beta-lactamase inhibitors, their use, and pharmaceutical composition

专利摘要:
BETA-LACTAMASE INHIBITORS, THEIR USE, AND PHARMACEUTICAL COMPOSITION. The invention relates to compounds and compositions that modulate the activity of beta-lactamases. In some embodiments, the compounds described herein inhibit beta-lactamase. In certain embodiments, the compounds described herein are useful in the treatment of bacterial infections.
公开号:BR112015013123B1
申请号:R112015013123-9
申请日:2013-12-05
公开日:2021-02-17
发明作者:Christopher J. Burns；Denis DAIGLE；Bin Liu；Daniel McGarry；Daniel C. Pevear；Robert E. Lee Trout
申请人:VenatoRx Pharmaceuticals, Inc.；
IPC主号:

专利说明:

[0001] [0001] This order claims the benefit of US Serial No. 61 / 734,900, filed on December 7, 2012, and US Serial No. 61 / 783,238, filed on March 14, 2013, each of which are hereby incorporated by reference in their entirety. FIELD OF THE INVENTION
[0002] [0002] The present invention relates to compounds containing boron, compositions, preparations and their use as inhibitors of beta-lactamase enzymes and as antibacterial agents. BACKGROUND OF THE INVENTION
[0003] [0003] Antibiotics are the most effective drugs for curing bacterial infectious diseases clinically. They have a wide market due to their advantages of good antibacterial effect with limited side effects. Among them, the beta-lactam class of antibiotics (eg, penicillins, cephalosporins and carbapenems) is widely used because it has a strong bactericidal effect and low toxicity.
[0004] [0004] To counteract the effectiveness of the various beta-lactams, bacteria have evolved to produce variants of beta-lactam deactivation enzymes called beta-lactamases, and with the ability to share this tool inter- and intra-species. These beta-lactamases are classified as "serine" or "metallo" based, respectively, on the presence of a serine or zinc key at the active site of the enzyme. The rapid spread of this bacterial resistance mechanism can severely limit options for beta-lactam treatment in the hospital and in the community. SUMMARY OF THE INVENTION
[0005] [0005] Compounds that modulate the activity of beta-lactamases are described herein. In some embodiments, the compounds described herein inhibit beta-lactamases. In certain embodiments, the compounds described herein are useful in the treatment of bacterial infections.
[0006] [0006] In one aspect, compounds of Formula I or Formula Ia, or their salts, solvates, polymorphs, stereoisomers, tautomers, prodrugs, metabolites, N-oxides or pharmaceutically acceptable isomers are provided here:
[0007] [0007] In some embodiments of a Formula I or Formula Ia compound, Ra, Rb, and Rc are independently selected from the group consisting of hydrogen, fluorine, chlorine, optionally substituted C1-C6 alkyl, optionally substituted C3-C6 cycloalkyl, -OH, -OR10, -NR4R5, and -SR10. In certain embodiments, Ra, Rb and Rc are independently hydrogen, fluorine, or chlorine. In the preferred embodiments, Ra, Rb, and Rc are hydrogen.
[0008] [0008] In some embodiments of a Formula I or Formula Ia compound, R3 is hydrogen, methyl, ethyl, propyl, butyl or isopropyl. In preferred embodiments, R3 is hydrogen.
[0009] [0009] In some embodiments of a Formula I or Formula Ia compound, X1 and X2 are -OH.
[0010] [0010] In some embodiments of a compound of Formula I or Formula Ia, Rd is hydrogen or C1-C4 alkyl. In preferred embodiments, Rd is hydrogen.
[0011] [0011] In some embodiments of a Formula I or Formula Ia compound, Z is> C = O or> SO2. In preferred embodiments, Z is> C = O.
[0012] [0012] In some embodiments of a Formula I or Formula Ia compound, L is -CR1R2- or = CR1-; M is -O-, -S-, -SO2-, or -N (R4) -; m is 0 or 1; and n is 1 or 2. In certain embodiments, L is a bond, -CR1R2-, or = CR1-; M is a bond or -O-; m is 0; and n is 1 or 2. In other embodiments, L is a bond or> C = O; M is a bond or -N (R4) -; and m and n are 0. In other embodiments, L is a bond; M is a bond; and m or n are 1. In some embodiments, L is -CR1R2-ou = CR1-; M is a bond; and m and n are 0. In certain embodiments, L is -CR1R2- or = CR1-; M is a bond; and m or n are 1.
[0013] [0013] In some embodiments of a Formula I or Formula Ia compound, CycA is selected from the group consisting of cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclopentene, cyclohexene, cyclohexene, heptene and cyclooctene, where the olefin functionality of cyclopentene, cyclohexene, cycloheptene and cyclooctene is not directly linked to an oxygen, sulfur or nitrogen substituent. In certain embodiments, CycA is cyclobutane, cyclopentane, cyclohexane or cyclohexene, in which the olefin functionality of cyclohexene is not directly linked to an oxygen, sulfur or nitrogen substituent. In other modalities, CycA is selected from the group consisting of octane bicycles [3.3.0], nonane bicycles [4.3.0], cis-decalines, trans-decalines, bicycles [2.1.1] hexane, bicycles [2.2.1] heptane, bicycles [2.2.2] octane, bicycles [3.2.2] nonane and bicycles [3.3.2] decane. In preferred embodiments, CycA is cyclobutane, cyclopentane, and cyclohexane. In some embodiments of a Formula I or Formula Ia compound, at least one Y is selected from the group of fluorine, chlorine, bromine, optionally substituted C1-C6 alkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted heterocycle, optionally substituted aryl, optionally substituted heteroaryl, = O, -OH, -OR10, -SR10, -NR4R5, - (CR6R7) vNR4R5, - (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4R5 (CR6R7) vR6, -NR4 (CR6R7) NR4 (CR6R7) vNR4R5 (CR6R7) vNR4R5, -O (CR6R7) vNR4R5, -S (O) o, 1,2 (CR6R7) vNR4R5, -N (R4) C (O) (CR6R7) vNR4R5, - (CR6R7) vN (R4) C (O) (CR6R7) vNR4R5, - (CR6R7) vNR4 (CR6R7) vNR4R5, -NR4 (CR6R7) vOR10, -NR4 (CR6R7) vS (O) o, 1,2R10, -C (O) NR4 (CR6R7) vNR4R5, -S (O) o, 1,2NR4 (CR6R7) vNR4R5, -NR5C (O) NR4 (CR6R7) vNR4R5, -OC (O) NR4 (CR6R7) vNR4R5, -NR5C (= NR7) NR4 (CR6R7) vNR4R5, -N (R4) C (= NR5) R6, - (CR6R7) vN (R4) C (= NR5) R6, -NR4 (CR6R7) vN (R4) C (= NR5) R6, -O (CR6R7) vN (R4) C (= NR5) R6, 0-S (O) o, 1,2 (CR6R7) vN (R4) C (= NR5) R6, - (CR6R7) vC (= NR5) NR4R5, -NR4 (CR6R7) vC (= NR5) NR4R5, -O (CR6R7) vC (= NR5) NR4R5, -S (O) o, 1,2 (CR6R7) vC (= NR 5) NR4R5, - (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4 (CR6R7) vN (R4) C (= NR5) NR4R5, -O (CR6R7) vN (R4) C (= NR5) NR4R5 , -S (O) o, 1,2 (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4C (= NR5) NR4C (= NR5) NR4R5, - (CR6R7) vC (= NR4) NR5 (= NR4) NR4R5, -NR4 (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4 (CR6R7) vNR4C (= NR4) NR4R5, -O (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -S (O) 0,1,2- (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4C (= NR5) NR4R5, -C (= NR4) NR4R5, -C (= NR4) NR4C ( O) R6, -NR4SO2R6, -NR4C (O) R6, -NR4C (= O) OR6, -C (O) NR4R5, - (CR6R7) vC (O) NR4R5, -SO2NR4R5, -Heteroaryl-NR4R5, -Heterocyclyl- NR4R5, -Heteroaryl-N (R4) C (= NR5) NR4R5, -Heterocyclyl-N (R4) C (= NR5) NR4R5, -N (R4) -Heteroaryl-NR4R5, -N (R4) -Heterocyclyl-NR4R5, - (CR6R7) vHeteroaryl-NR4R5, - (CR6R7) vHeterocyclyl-NR4R5, - (CR6R7) vHeteroaryl-N (R4) (= NR5) NR4R5, - (CR6R7) vHeterocyclyl-N (R4) C (= NR5) NR4R5, - NR4R5 (CR6R7) vHeterocyclyl-C (= NR5) NR4R5 - (CR6R7) vHeteroaryl, - (CR6R7) vHeterocyclyl, -O-Heteroaryl, -O-Heterocyclyl, -NR4 (CR6R7) vHeteroaryl, -NR4 (CR6R7) (CR6R7) vHeteroaryl, -O (CR6R7) vHeterocyclyl, -NR4 (CR6R7) vNR5-Het eroaryl, -NR4 (CR6R7) vNR5-Heterocyclyl, -O (CR6R7) vNR5-Heteroaryl, -O (CR6R7) vNR5-Heterocyclyl, -O (CR6R7) vO-Heterocyclyl, -NR4R5R9 + Q-, - (CR6R7) Q-, -NR4 (CR6R7) vNR4R5R9 + Q-, -NR4R9 + (CR6R7) vNR4R5R9 + Q-2, - (CR6R7) v (T) + Q-, and -O (CR6R7) vNR4R5R9 + Q-; on what: each T is independently selected from the group consisting of pyridine-1-yl, pyrimidin-1-yl, and thiazol-3-yl; each Q is independently a pharmaceutically acceptable counterion; and each v is independently 1, 2, 3, or 4; or Y taken together with the carbon atom to which it is attached forms an optionally substituted spiro-carbocycle or optionally substituted spiro-heterocycle; or two Y taken together with the carbon atoms to which they are attached form an optionally substituted carbocycle or an optionally substituted heterocycle; each of R6 and R7 is independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, optionally substituted C1-C6 alkyl, optionally substituted alkoxyalkyl, optionally substituted hydroxyalkyl, optionally substituted C3-C6 cycloalkyl, -OH, -OR10, -SR10, -NR4R5, -NR4C (O) R5, -NR4C (O) OR5, -NR4C (O) NR5, -C (O) OR5, -C (O) NR4R5, -C (N = R5) NR4R5 - NR4SO2R5, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; or R6 and R7 taken together form an oxo, oxime, or an optionally substituted carbocycle or a heterocycle optionally substituted with the carbon to which they are attached; each R9 is independently optionally substituted C1-C6 alkyl. In some embodiments, at least one Y comprises 1 to 6 basic nitrogen atoms. In some embodiments, at least one Y comprises 1, 2 or 3 basic nitrogen atoms. In some embodiments, at least one Y comprises 2 basic nitrogen atoms.
[0014] [0014] In some embodiments of a Formula I or Formula Ia compound, at least one Y is selected from the group consisting of fluorine, chlorine, optionally substituted C1-C6 alkyl, = O, -OH, -OR10, -NR4R5, - (CR6R7) vNR4R5, -NR4 (CR6R7) vNR4R5, - (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4R5 (CR6R7) vR6, -NR4R5 (CR6R7) vHeterocycly-C (= NR5) = NR4) NR4R5, -NR4 (CR6R7) vNR4R5 (CR6R7) vNR4R5, -O (CR6R7) vNR4R5, -N (R4) C (O) (CR6R7) vNR4R5, - (CR6R7) vN (R4) C (O) ( CR6R7) vNR4R5, -C (O) NR4 (CR6R7) vNR4R5, -S (O) 0,1,2NR4 (CR6R7) vNR4R5, -NR5C (O) NR4 (CR6R7) vNR4R5, -OC (O) NR4 (CR6R7) vNR4R5, -NR5C (= NR7) NR4 (CR6R7) vNR4R5, -N (R4) C (= NR5) R6, - (CR6R7) vN (R4) C (= NR5) R6, -NR4 (CR6R7) vN (R4) C (= NR5) R6, -O (CR6R7) vN (R4) C (= NR5) R6, - (CR6R7) vC (= NR5) NR4R5, -NR4 (CR6R7) vC (= NR5) NR4R5, -O (CR6R7 ) vC (= NR5) NR4R5, - (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4 (CR6R7) vN (R4) C (= NR5) NR4R5, -O (CR6R7) vN (R4) C ( = NR5) NR4R5, -NR4C (= NR5) NR4C (= NR5) NR4R5, - (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4 (CR6R7) v (= NR4) NR5C (= NR4) NR4R5 , -O (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4C (= NR5) NR4R5, -C (= NR4) NR4R5, -C (= NR4) NR4C (O) R6, -NR4SO2R6, -NR4C (O) R6, -NR4C (= O) OR6, -C (O) NR4R5, - (CR6R7) vC (O ) NR4R5, -Heteroaryl-NR4R5, -Heterocyclyl-NR4R5, -Heteroaryl-N (R4) C (= NR5) NR4R5, -Heterocyclyl-N (R4) C (= NR5) NR4R5, -N (R4) -Heteroaryl-NR4R5 , -N (R4) -Heterocyclyl-NR4R5, - (CR6R7) vHeteroaryl-NR4R5, - (CR6R7) vHeterocyclyl-NR4R5, - (CR6R7) vHeteroaryl-N (R4) C (= NR5) NR4R5, - (CR6R7) vHeterocycle N (R4) C (= NR5) NR4R5, - (CR6R7) vHeteroaryl, - (CR6R7) vHeterocyclyl, -O-Heteroaryl, -O-Heterocyclyl, -NR4 (CR6R7) vHeteroaryl, -NR4 (CR6R7) vHeterocyclyl, -O ( CR6R7) vHeteroaryl, -O (CR6R7) vHeterocyclyl, and -O (CR6R7) vO-Heterocyclyl. In certain embodiments, at least one Y is selected from the group consisting of fluorine, optionally substituted C1-C6 alkyl, -OH, -NR4R5, - (CR6R7) vNR4R5, - (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4R5 (CR6R7) vR6, -NR4R5 (CR6R7) vHeterocyclyl-C (= NR5) NR4R5, -NR4 (CR6R7) vNR4C (= NR4) NR4R5, -NR4 (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4 (CR6R7) ) vNR4R5, -C (O) NR4 (CR6R7) vNR4R5, -NR5C (O) NR4 (CR6R7) vNR4R5, -NR5C (= NR7) NR4 (CR6R7) vNR4R5, -N (R4) C (= NR5) R6, - (CR6R7) vN (R4) C (= NR5) R6, -NR4 (CR6R7) vN (R4) C (= NR5) R6, - (CR6R7) vC (= NR5) NR4R5, -NR4 (CR6R7) vC (= NR5 ) NR4R5, - (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4 (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4C (= NR5) NR4C (= NR5) NR4R5, - (CR6R7 ) vC (= NR4) NR5C (= NR4) NR4R5, -NR4 (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4C (= NR5) NR4R5, -C (= NR4) NR4R5, -C (= NR4) NR4C (O) R6, -NR4C (O) R6, - (CR6R7) vC (O) NR4R5, -Heterocyclyl-NR4R5, -Heterocyclyl-N (R4) C (= NR5) NR4R5, -N (R4) - Heterocyclyl-NR4R5, - (CR6R7) vHeterocyclyl-NR4R5, - (CR6R7) vHeterocyclyl-N (R4) C (= NR5) NR4R5, - (CR6R7) vHeterocyclyl, and -NR4 (CR6R7) vHeterocyclyl. In other embodiments, at least one Y is selected from the group consisting of -Heteroaryl-NR4R5, -Heterocyclyl-NR4R5, -Heteroaryl-N (R4) C (= NR5) NR4R5, -Heterocyclyl-N (R4) C (= NR5 ) NR4R5, -N (R4) -Heteroaryl-NR4R5, -N (R4) -Heterocyclyl-NR4R5, -Heteroaryl-C (= NR5) NR4R5, -Heterocyclyl-C (= NR5) NR4R5, - (CR6R7) vHeteroaril-NR4R5 , - (CR6R7) vHeterocyclyl-NR4R5, - (CR6R7) vHeteroaryl-N (R4) C (= NR5) NR4R5, and - (CR6R7) vHeterocyclyl-N (R4) C (= NR5) NR4R5. In preferred embodiments, at least one Y is selected from the group consisting of -NR4R5, -NR4C (= NR5) NR4R5, -C (= NR4) NR4R5, -N (R4) C (= NR5) R6, - (CR6R7) vNR4R5, - (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4 (CR6R7) vNR4R5, -NR4 (CR6R7) vOR10, - (CR6R7) vNR4 (CR6R7) vNR4R5, NR5C (= NR5) NR4 (NR5) NR4 vNR4R5, -NR4 (CR6R7) vN (R4) C (= NR5) NR4R5, -NR5C (O) CR6 (NR4R5) (CR6R7) vNR4R5, - (CR6R7) vC (= NR5) NR4R5, - (CR6R7) vN (R4 ) C (O) (CR6R7) vNR4R5, -C (= NR4) NR4C (O) R6, -NR4 (CR6R7) vHeteroaryl, and -O (CR6R7) vNR4R5.
[0015] [0015] In some modalities, p is 0, 1, 2, 3, or 4. In certain modalities, p is 1 or 2. In some modalities, p is 1.
[0016] [0016] In some embodiments of a Formula I or Formula Ia compound, R4 and R5 are independently selected from the group consisting of hydrogen, -OH, C1-C6 optionally substituted alkyl, optionally substituted alkoxyalkyl, optionally substituted hydroxyalkyl, and optionally heterocyclyl replaced. In preferred embodiments, R4 and R5 are independently hydrogen or optionally substituted C1-C6 alkyl.
[0017] [0017] In some embodiments of a Formula I or Formula Ia compound, R6 and R7 are independently selected from the group consisting of hydrogen, optionally substituted C1-C6 alkyl, -OH, -NR4R5, and optionally substituted heterocyclyl, or R6 and R7 taken together form a heterocycle optionally substituted with the carbon to which they are attached. In preferred embodiments, R6 and R7 are independently hydrogen, fluorine, or optionally substituted C1-C6 alkyl. In some modalities,
[0018] [0018] In certain embodiments of a Formula I or Formula Ia compound, the compound is selected from the group represented by the following structures:
[0019] In another aspect, pharmaceutical compositions comprising a compound of Formula I or Formula Ia as described herein, or a salt, solvate, polymorph, stereoisomer, tautomer, prodrug, metabolite, N-oxide or isomer thereof, are provided herein. and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition further comprises a beta-lactam antibiotic. In certain embodiments, the beta-lactam antibiotic is a penicillin, cephalosporin, carbapenem, monobactam, bridged monobactam, or a combination of these.
[0020] [0020] In another aspect, methods of treating a bacterial infection in an individual are provided herein, comprising administering to the individual a pharmaceutical composition as described herein, optionally in combination with a beta-lactam antibiotic. In certain embodiments, methods of treating a bacterial infection in an individual comprise administering to the individual a pharmaceutical composition as described herein in combination with a beta-lactam antibiotic. INCORPORATION BY REFERENCE
[0021] [0021] All publications, patents and patent applications mentioned in this specification are incorporated herein by reference to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference. DETAILED DESCRIPTION OF THE INVENTION
[0022] [0022] Beta-lactamases are typically grouped into 4 classes: classes Ambler A, B, C, and D, based on their amino acid sequences. Enzymes in classes A, C and D are serine beta-lactamases of active site, whereas enzymes in class B are dependent on Zn. The most recent cephalosporins and carbapenens have been partially developed based on their ability to evade the effect of deactivating beta-lactamase variants based on early serine. However, a recent wave of new versions of serine-based beta-lactamases - for example, Class A Extended-Spectrum Beta-Lactamase (ESBL) enzymes, class A carbapenemases (eg KPC-2), class cephalosporinases C-mediated chromosomes and plasmids (AmpC, CMY, etc.), and class D oxacillinases - as well as class B metallo-beta-lactamases (eg, VIM, NDM) began to diminish the utility of the antibiotic family of beta-lactam, including the latest generation beta-lactam drugs, leading to a serious medical condition. In fact, the number of cataloged serine-based beta-lactamases exploded from less than ten in the 1970s to more than 750 variants (see, for example, Jacoby & Bush, “Amino Acid Sequences for TEM, SHV and OXA Extended- Spectrum and Inhibitor Resistant β-Lactamases ”, on the Lahey Clinic website).
[0023] [0023] Commercially available beta-lactamase inhibitors (clavulanic acid, sulbactam, tazobactam) were developed to treat beta-lactamases that were clinically relevant in the 1970s and 1980s (for example, penicillinases). These beta-lactamase inhibitors are weakly active against the diversity of beta-lactamase enzymes (based on both serine and metallo) that now appear clinically. In addition, these enzyme inhibitors are only available as fixed combinations with penicillin derivatives. No combination with cephalosporins (or carbapene-mos) is clinically available. This fact, combined with the increased use of more recent generation cephalosporins and carbapenens, is driving the selection and diffusion of new variants of beta-lactamase (ESBLs, carbapenemases, chromosomes and Class C, Class D plasmid-mediated oxacillinases, etc.). ). While maintaining good inhibitory activity against ESBLs, inherited beta-lactamase inhibitors are largely ineffective against the new Class A and Class B carbapenemases, against the plasmid-mediated Class C chromosome and cephalosporinases and against many of the Class D oxacillinases.
[0024] [0024] To address this growing therapeutic vulnerability, and because there are three main molecular classes of serine-based beta-lactamases, and one main class of metallo-beta-lactamases, and each of these classes contains significant numbers of variants of beta- lactamase, we identified an approach for the development of new beta-lactamase inhibitors with broad spectrum functionality. In particular, we have identified an approach to the development of compounds that are active against both beta-lactamase enzymes based on both serine and metal. The compounds of the present invention demonstrate potent activity in all four major classes of beta-lactamases.
[0025] [0025] The present invention is directed to certain boron-based compounds (boronic acids and cyclic boronic acid esters), which are beta-lactamase inhibitors and antibacterial compounds. The compounds and their pharmaceutically acceptable salts are useful alone and in combination with beta-lactam antibiotics for the treatment of bacterial infections, particularly antibiotic-resistant bacterial infections. Some modalities include compounds, compositions, pharmaceutical compositions, their use and preparation. Definitions
[0026] [0026] In the description that follows, certain specific details are presented in order to provide a complete understanding of the various modalities. However, a person skilled in the art will understand that the invention can be practiced without these details. In other cases, well-known structures have not been presented or described in detail to avoid unnecessary obscure descriptions of the modalities. Unless the context otherwise requires, throughout the specification and claims that follow, the word "understand" and its variations, such as "understand" and "understanding", must be interpreted in an inclusive open sense , that is, as "including, but not limited to, these". In addition, the topics provided herein are for convenience only and not to interpret the scope or meaning of the claimed invention.
[0027] [0027] The reference throughout this specification to "a modality" or "an embodiment", means that a particular aspect, structure or characteristic described in connection with the modality is included in at least one modality. Thus, the appearances of the phrases "in one modality" or "in one embodiment" in various places throughout this specification are not necessarily referring entirely to the same modality. In addition, the particular aspects, structures or characteristics can be combined in any suitable way in one or more modalities. Likewise, as used in this specification and the appended claims, the singular forms "one", "one", "o" and "a" include references in the plural, unless the content clearly dictates otherwise. It should also be noted that the term "or" is generally used in its sense, including "and / or", unless the content clearly dictates otherwise.
[0028] [0028] The term "antibiotic" refers to a compound or composition that decreases the viability of a microorganism, or that inhibits the growth or proliferation of a microorganism. The phrase "inhibits growth or proliferation" means increasing the generation time (that is, the time required for the bacterial cell to divide or for the population to fold) at least around 2 times. Preferred antibiotics are those that can increase the generation time by at least about 10 times or more (for example, at least about 100 times or even indefinitely, as in total cell death). As used in this description, an antibiotic is further intended to include an antimicrobial, bacteriostatic or bactericidal agent. Examples of antibiotics suitable for use in connection with the present invention include penicillins, cephalosporins and carbapenems.
[0029] [0029] The term "β-lactam antibiotic" refers to a compound with antibiotic properties that contains a β-lactam functionality. Non-limiting examples of β-lactam antibiotics useful with respect to the invention include penicillins, cephalosporins, penems, carbapenems and monobactams.
[0030] [0030] The term "β-lactamase" means a protein capable of inactivating a β-lactam antibiotic. Β-lactamase can be an enzyme that catalyzes the hydrolysis of the β-lactam ring of an β-lactam antibiotic. Of particular interest here are microbial β-lactamases. Β-lactamase can be, for example, a β-lactamase serine or a metallo-lactamase. Β-lactamases of interest include those described on a continuous website that monitors the beta-lactamase nomenclature (www.lahey.org) and in Bush, K. and G. A. Jacoby. 2010. An updated functional classification of β-lactamases. Antimicrob. Chemother Agents. 54: 969-976. The β-lactamases of particular interest here include the β-lactamases found in bacteria such as the class A β-lactamases including the subclasses SHV, CTX-M and KPC, the class B β-lactamases such as VIM, the β- class C lactamases (mediated by both chromosome and plasmid), and class D β-lactamases. The term "β-lactamase inhibitor" refers to a compound that is capable of inhibiting β-lactamase activity. Inhibition of β-lactamase activity means inhibition of the activity of a class A, B, C or D β-lactamase. For antimicrobial applications, inhibition at a 50% inhibitory concentration is preferably achieved at or below about 100 micrograms / ml, or at or below about 50 micrograms / ml, or at or below about 25 micrograms / ml. The terms β-lactamases of "class A", "class B", "class C" and "class D" are understood by those skilled in the art and are described in Bush, K. and G. A. Jacoby. 2010. An updated functional classification of β-lactamases. Antimicrob. Chemother Agents. 54: 969-976.
[0031] [0031] The terms below, as used herein, have the following meanings, unless otherwise indicated.
[0032] [0032] "Amino" refers to the radical -NH2.
[0033] [0033] "Cyan" or "nitrile" refers to the radical -CN.
[0034] [0034] "Hydroxy" or "hydroxyl" refers to the radical -OH.
[0035] [0035] "Nitro" refers to the radical -NO2.
[0036] [0036] "Oxo" refers to the substituent = O.
[0037] [0037] "Oxima" refers to the substituent = N-OH.
[0038] [0038] "Thioxus" refers to the substituent = S.
[0039] [0039] "Alkyl" refers to an optionally substituted straight chain or optionally substituted saturated hydrocarbon monoradical having from one to about ten carbon atoms, more preferably from one to six carbon atoms, where one carbon sp3-hybridized alkyl residue is linked to the rest of the molecule by a single bond. Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1- butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2- pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, tert-amyl and hexyl, and larger alkyl groups, such as heptyl, octyl and the like. When a numerical range such as "C1-C6 alkyl" or "C1-6 alkyl" appears here, it means that the alkyl group can consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms , 5 carbon atoms or 6 carbon atoms, although this definition also covers the occurrence of the term "alkyl" where no numerical range is designated. Unless otherwise specifically mentioned in the specification, an alkyl group can be optionally substituted as described below, for example, with oxo, amino, nitrile, nitro, hydroxyl, alkyl, alkylene, alkynyl, alkoxy, aryl, cycloalkyl , heterocyclyl, heteroaryl, and the like.
[0040] [0040] "Alkenyl" refers to an optionally substituted straight chain hydrocarbon or optionally substituted branched chain having one or more carbon-carbon double bonds and having two to about ten carbon atoms, more preferably two to about of six carbon atoms, in which a sp2-hybridized carbon of the alkenyl residue is attached to the rest of the molecule by a single bond. The group can be in the cis or trans conformation around the double bonds, and should be understood to include both isomers. Examples include, but are not limited to, ethylene (-CH = CH2), 1-propenyl (-CH2CH = CH2), isopropenyl [-C (CH3) = CH2], butenyl, 1,3-butadienyl and the like. Whenever a numerical range such as "C2-C6 alkenyl" or "C2-6 alkenyl" appears here, it means that the alkenyl group can consist of two carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 atoms of carbon or 6 carbon atoms, although this definition also covers the occurrence of the term "alkenyl", where no numerical range is designated.
[0041] [0041] "Alquinyl" refers to an optionally substituted straight chain or optionally substituted branched hydrocarbon mono-radical having one or more carbon-carbon triple bonds and having two to about ten carbon atoms, more preferably two at about six carbon atoms. Examples include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, 1,3-butadinyl and the like. When a numerical range such as "C2-C6 alkynyl" or "C2-6 alkynyl" appears here, it means that the alkynyl group can consist of two carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms, although this definition also covers the occurrence of the term "alkynyl" where no numerical range is designated.
[0042] [0042] "Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain. Unless specifically mentioned otherwise in the specification, an alkylene group can be optionally substituted as described below.
[0043] [0043] "Aloxy" refers to a radical of the formula ORa, where Ra is an alkyl radical as defined. Unless specifically mentioned otherwise in the specification, an alkoxy group can be optionally substituted as described below.
[0044] [0044] "Arila" refers to a radical derived from a hydrocarbon ring system comprising hydrogen, from 6 to 30 carbon atoms and at least one aromatic ring. The aryl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems. Aryl radicals include, but are not limited to, aryl radicals derived from the aceantrile-no, acenaftylene, acefenantrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s- hydrocarbon ring systems indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene and triphenylene. Unless specifically mentioned otherwise in the specification, the term "aryl" or the prefix "ar-" (as in "aralkyl") is intended to include the aryl radicals that are optionally substituted.
[0045] [0045] "Cycloalkyl" or "carbocycle" refers to a stable non-aromatic, monocyclic or polycyclic carbocyclic ring, which may include fused or bridged ring systems, which is saturated or unsaturated. Representative cycloalkyls or carbocycles include, but are not limited to, cycloalkyls having three to fifteen carbon atoms, three to ten carbon atoms, three to eight carbon atoms, three to six carbon atoms, three to five carbon atoms, or three to four carbon atoms. Cycloalkyls or monocyclic carbocycles include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl and cyclooctyl. Polycyclic cycloalkyls or carbocycles include, for example, adamantyl, norbornyl, decalinyl, bicyclo [3.3.0] octane, bicyclo [4.3.0] nonane, cis-decalin, trans-decaline, bicycle [2.1.1] hexane, bicyclo 2.2.1] heptane, bicycles [2.2.2] octane, bicycles [3.2.2] nonane and bicycles [3.3.2] decane, and 7,7-dimethyl-bicycles [2.2.1] heptanyl. Unless otherwise specifically mentioned in the specification, a cycloalkyl or carbocycle group can be optionally substituted. Illustrative examples of cycloalkyl groups include, but are not limited to, the following components:
[0046] [0046] "Aralkyl" means a radical of - (alkylene) -R where R is aryl as defined above.
[0047] [0047] "Cycloalkylalkyl" means a radical of - (alkylene) -R where R is cycloalkyl as defined above; for example, cyclopropylmethyl, cyclobutylmethyl, cyclopentylethyl or cyclohexylmethyl, and the like.
[0048] [0048] "Fused" refers to any ring structure described herein that is fused to an existing ring structure. When the fused ring is a heterocyclyl ring or a heteroaryl ring, any carbon atom in the existing ring structure that becomes part of the fused heterocyclyl ring or the fused heteroaryl ring can be replaced with a nitrogen atom.
[0049] [0049] "Halo" or "halogen" refers to bromine, chlorine, fluorine or iodine.
[0050] [0050] "Haloalkyl" refers to an alkyl radical, as defined above, which is replaced by one or more halo radicals, as defined above, for example, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2- trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like. Unless specifically mentioned otherwise in the specification, a haloalkyl group can be optionally substituted.
[0051] [0051] "Haloalkoxy" similarly refers to a radical of the formula -ORa, where Ra is a haloalkyl radical as defined. Unless specifically mentioned otherwise in the specification, a haloalkoxy group can be optionally substituted as described below.
[0052] [0052] "Heterocycloalkyl" or "heterocyclyl" or "heterocyclic ring" or "heterocycle" refers to a stable 3- to 24-membered non-aromatic ring radical comprising from 2 to 23 carbon atoms and from 1 to 8 heteroatoms selected from the group consisting of nitrogen, oxygen, phosphorus and sulfur. Unless specifically mentioned otherwise in the specification, the heterocyclyl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclyl radical can be optionally oxidized; the nitrogen atom can be optionally quaternized; and the heterocyclyl radical can be partially or completely saturated. Examples of such heterocyclyl radicals include, but are not limited to, azetidinyl, dioxolanyl, thienyl [1,3] dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octa-hydroindolyl, octahydroxyindole , 2-oxopiperi-dinila, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, tritianyl, tetrahydropyrilyl, thiororphinin, thiororphinin, thiororphinin -tiomorpholinyl, 12-crown-4, 15-crown-5, 18-crown-6, 21-crown-7, aza-18-crown-6, diaza-18-crown-6, aza-21-crown-7 , and diaza-21-crown-7. Unless otherwise specifically mentioned in the specification, a heterocyclyl group can be optionally substituted. Illustrative examples of heterocycloalkyl groups, also referred to as non-aromatic heterocycles, include:
[0053] [0053] "Heteroaryl" refers to a radical of the 5- to 14-membered ring system comprising hydrogen atoms, from one to thirteen carbon atoms, from one to six heteroatoms selected from the group consisting of nitrogen, oxygen, phosphorus and sulfur, and at least one aromatic ring. For the purposes of this invention, the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl radical can be optionally oxidized; the nitrogen atom can be optionally quaternized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo [o] [1,4] dioxepinyl, 1,4-benzodoxylanyl, benzodoxylanyl, benzodoxanil, benzone , benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo [4,6] imidazo [1,2-a] pyridinyl, carbazolyl, cinnolinyl, dibenzoturanyl, furibyl, furibyl, furonol imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxopyridinyl, 1-oxopyridinyl, 1-oxopyridinyl, 1 phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyr idazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl and thiophenyl (i.e., thienyl). Unless specifically mentioned otherwise in the specification, a heteroaryl group can be optionally substituted.
[0054] [0054] All of the above groups can be replaced or not replaced. The term "substituted" as used herein means any of the above groups (for example, alkyl, alkylene, alkoxy, aryl, cycloalkyl, haloalkyl, heterocyclyl and / or heteroaryl), it can be further functionalized in which at least one hydrogen atom is replaced by a bond to a non-hydrogen atom substituent. Unless specifically mentioned in the specification, a substituted group may include one or more substituents selected from: oxo, amino, -CO2H, nitrile, nitro, hydroxyl, thiooxy, alkyl, alkylene, alkoxy, aryl, cycloalkyl, heterocyclyl, heteroaryl , dialkylamines, arylamines, alkylarylamines, diarylamines, trialkylammonium (-N + R3), N-oxides, imides and enamines; a silicon atom in groups such as trialkylsilyl groups, dialkylarylsilyl groups, alkyldarylsilyl groups, triarylsilyl or perfluoroalkyl or perfluoroalkoxy groups, for example, trifluoromethyl or trifluoromethoxy. "Substituted" also means any of the above groups in which one or more hydrogen atoms are replaced by a higher order bond (for example, a double or triple bond) to a hetero atom such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and nitrogen in groups such as imines, oximes, hydrazones and nitriles. For example, "substituted" includes any of the above groups in which one or more hydrogen atoms are replaced by -NH2, -NRgC (= O) NRgRh, -NRgC (= O) ORh, -NRgSO2Rh, -OC (= O ) NRgRh, -ORg, -SRg, -SORg, -SO2Rg, -OSO2Rg, -SO2ORg, = NSO2Rg, and -SO2NRgRh. In the foregoing, Rg and Rh are the same or different and independently hydrogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl and / or heteroaryl and / or heteroaryl . In addition, each of the above substituents can also optionally be substituted with one or more of the above substituents. In addition, any of the groups above can be substituted to include one or more of internal atoms of oxygen, sulfur or nitrogen. For example, an alkyl group can be replaced with one or more internal oxygen atoms to form an ether or polyether group. Likewise, an alkyl group can be substituted with one or more internal sulfur atoms to form a thioether, disulfide, etc.
[0055] [0055] The term "optional" or "optionally" means that the event or circumstance subsequently described may or may not occur, and that the description includes cases where said event or circumstance occurs and cases in which it does not. For example, "optionally substituted alkyl" means "alkyl" or "substituted alkyl", as defined above. In addition, an optionally substituted group can be unsubstituted (for example, -CH2CH3), fully substituted (for example, -CF2CF3), monosubstituted (for example, -CH2CH2F) or substituted at a level for any part between fully substituted and mono -substituted (for example, -CH2CHF2, -CH2CF3, -CF2CH3, -CFHCHF2, etc.). It will be understood by those skilled in the art in relation to any group containing one or more substituents that such groups are not intended to introduce any substitution or substitution patterns (for example, substituted alkyl includes optionally substituted cycloalkyl groups, which in turn are defined as including optionally substituted alkyl groups, potentially ad infinitum) that are sterically impractical and / or synthetically non-viable. Thus, any substituents described should generally be understood to have a maximum molecular weight of about 1,000 Daltons, and more typically, up to about 500 Daltons.
[0056] [0056] An "effective amount" or "therapeutically effective amount" refers to an amount of a compound administered to a mammalian individual, either as a single dose or as part of a series of doses, which is effective to produce a therapeutic effect wanted.
[0057] [0057] "Treatment" of an individual (for example, a mammal, such as a human being) or a cell is any type of intervention used in an attempt to alter the natural course of the individual or cell. In some embodiments, treatment includes administration of a pharmaceutical composition, subsequent to the onset of a pathological event or contact with an etiologic agent, and includes stabilizing the condition (for example, the condition does not get worse) or relieving the condition. In other embodiments, treatment also includes prophylactic treatment (for example, administration of a composition described herein when an individual is suspected of suffering from a bacterial infection).
[0058] [0058] A "tautomer" refers to a change in protons from one atom of a molecule to another atom of the same molecule. The compounds presented herein can exist as tautomers. Tautomers are compounds that are interconvertible by the migration of a hydrogen atom, accompanied by a transition of a single bond and an adjacent double bond. In the bonding arrangements where tautomerization is possible, a chemical balance of the tautomers will exist. All tautomeric forms of the compounds disclosed herein are contemplated. The exact ratio of tautomers depends on several factors, including temperature, solvent and pH. Some examples of tautomeric interconversions include:
[0059] [0059] A "metabolite" of a compound disclosed herein is a derivative of that compound that is formed when the compound is metabolized. The term "active metabolite" refers to a biologically active derivative of a compound that is formed when the compound is metabolized. The term "metabolized", as used herein, refers to the sum of processes (including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes, such as oxidation reactions) through which a particular substance is altered by an organism. Thus, enzymes can produce specific structural changes in a compound. For example, cytochrome P450 catalyzes a variety of oxidative and reductive reactions, while uridine diphosphate glucuronyl transferases catalyze the transfer of an activated glucuronic acid molecule to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines and sulfhydryl-free groups. Further information on metabolism can be obtained from The Pharmacological Basis of Therapeutics, 9th Edition, McGraw-Hill (1996). The metabolites of the compounds disclosed herein can be identified by administering compounds to a host and analyzing tissue samples from the host, or by incubating the compounds with liver cells in vitro and analyzing the resulting compounds. Both methods are well known in the art. In some embodiments, the metabolites of a compound are formed by oxidative processes and correspond to the corresponding hydroxy-containing compound. In some embodiments, a compound is metabolized to pharmacologically active metabolites. Compounds
[0060] [0060] Compounds that modulate beta-lactamase activity are described herein. In some embodiments, the compounds described herein inhibit beta-lactamase. In certain embodiments, the compounds described herein are useful in the treatment of bacterial infections. In some embodiments, bacterial infection is an upper or lower respiratory tract infection, an urinary tract infection, an intra-abdominal infection, or a skin infection.
[0061] [0061] In one aspect, compounds of Formula I or Formula Ia, or their salts, solvates, polymorphs, stereoisomers, tautomers, prodrugs, metabolites, N-oxides or pharmaceutically acceptable isomers are provided here:
[0062] [0062] In some embodiments of a Formula I or Formula Ia compound, Ra, Rb, and Rc are independently selected from the group consisting of hydrogen, fluorine, chlorine, optionally substituted C1-C6 alkyl, optionally substituted C3-C6 cycloalkyl, -OH, -OR10, -NR4R5, and -SR10. In certain embodiments, Ra, Rb, and Rc are independently hydrogen, fluorine, or chlorine. In the preferred embodiments, Ra, Rb, and Rc are hydrogen.
[0063] [0063] In some embodiments of a Formula I or Formula Ia compound, R3 is hydrogen, methyl, ethyl, propyl, butyl, or isopropyl. In preferred embodiments, R3 is hydrogen.
[0064] [0064] In some embodiments of a compound of Formula I or Formula Ia, X1 and X2 are -OH.
[0065] [0065] In some embodiments of a compound of Formula I or Formula Ia, Rd is hydrogen or C1-C4-alkyl. In some embodiments, Rd is methyl. In preferred embodiments, Rd is hydrogen.
[0066] [0066] In some embodiments of a Formula I or Formula Ia compound, Z is Z is> C = O or> SO2. In preferred embodiments, Z is> C = O.
[0067] [0067] In some embodiments of a Formula I or Formula Ia compound, L is -CR1R2- or = CR1-. In certain embodiments, L is a bond. In some embodiments of a Formula I or Formula Ia compound, M is -O-, -S-, -SO2-, or -N (R4) -. In certain embodiments, M is a bond or -O-. In other modalities, M is a link. In some embodiments of a Formula I or Formula Ia compound, m is 0 or 1. In certain embodiments, m is 0. In other embodiments, m is 1. In some embodiments of a Formula I or Formula Ia compound, n is 1 or 2. In certain modalities, n is 1. In other modalities, n is 0. In other modalities, n is 2. In some modalities, men are 0. In certain modalities, m or n are 1.
[0068] [0068] In some embodiments of a Formula I or Formula Ia compound, L is -CR1R2- or = CR1-; M is -O-, -S-, -SO2-, or -N (R4) -; m is 0 or 1; and n is 1 or 2. In certain embodiments, L is a bond, -CR1R2-, or = CR1-; M is a bond or -O-; m is 0; and n is 1 or 2. In other embodiments, L is a bond or> C = O; M is a bond or -N (R4) -; and m and n are 0. In some embodiments, L is> C = O; M is -N (R4) -; m and n are 0. In certain embodiments, L is a bond; M is a bond; m and n are 0. In other embodiments, L is a bond; M is a bond; m or n are 1. In some embodiments, L is -CR1R2-ou = CR1-; M is a bond; m and n are 0. In certain embodiments, L is -CR1R2- or = CR1-; M is a bond; m or n is 1.
[0069] [0069] In some embodiments of a Formula I or Formula Ia compound, CycA is selected from the group consisting of cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclopentene, cyclohexene, cyclohexene, heptene and cyclooctene, where the olefin functionality of cyclopentene, cyclohexene, cycloheptene and cyclooctene is not directly linked to an oxygen, sulfur or nitrogen substituent. In certain embodiments, CycA is cyclobutane, cyclopentane, cyclohexane or cyclohexene, in which the olefin functionality of cyclohexene is not directly linked to an oxygen, sulfur or nitrogen substituent. In other modalities, CycA is selected from the group consisting of octane bicycles [3.3.0], nonane bicycles [4.3.0], cis-decalines, trans-decalines, bicycles [2.1.1] hexane, bicycles [2.2.1] heptane, octane bicycle [2.2.2], nonane bicycle [3.2.2], and decane bicycle [3.3.2]. In some embodiments, CycA is cyclopentane. In preferred embodiments, CycA is cyclohexane. In some embodiments, CycA is cyclohexane covalently linked to a Y and L; said covalent bonds in the 1,4-trans arrangement.
[0070] [0070] In some embodiments of a Formula I or Formula Ia compound, each Y is selected from the group consisting of fluorine, chlorine, bromine, optionally substituted C1-C6 alkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted heterocycle, aryl optionally substituted, optionally substituted heteroaryl, = O, -OH, -OR10, -SR10, -NR4R5, - (CR6R7) vNR4R5, - (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4R5 (CR6R7) vR6, -NR4 vHeterocyclyl-C (= NR5) NR4R5, -NR4 (CR6R7) vNR4C (= NR4) NR4R5, -NR4 (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4 (CR6R7) vNR4R5, -O (CR6R7) ) o, i, 2 (CR6R7) vNR4R5, -N (R4) C (O) (CR6R7) vNR4R5, - (CR6R7) vN (R4) C (O) (CR6R7) vNR4R5, - (CR6R7) vNR4 (CR6R7) vNR4R5, -NR4 (CR6R7) vOR10, -NR4 (CR6R7) vS (O) 0,1,2R10, -C (O) NR4 (CR6R7) vNR4R5, -S (O) 0,1,2NR4 (CR6R7) vNR4R5, -NR5C (O) NR4 (CR6R7) vNR4R5, -OC (O) NR4 (CR6R7) vNR4R5, -NR5C (= NR7) NR4 (CR6R7) vNR4R5, -N (R4) C (= NR5) R6, - (CR6R7) vN (R4) C (= NR5) R6, -NR4 (CR6R7) vN (R4) C (= NR5) R6, -O (CR6R7) vN (R4) C (= NR5) R6, -S (O) o, 1.2 (CR6R7) vN (R4) C (= NR5) R6, - (CR6R7) vC (= NR5) NR4R5, -NR4 (CR6R7) vC (= NR5) NR4R5, -O (CR6R7) vC (= NR5) NR4R5, -S (O) o, 1,2 (CR6R7) vC (= NR5) NR4R5, - (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4 (CR6R7) vN (R4) C (= NR5) NR4R5, -O (CR6R7) vN (R4) C (= NR5) NR4R5, -S (O) o, 1.2 (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4C (= NR5) NR4C (= NR5) NR4R5, - (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4 (C R6R7) vC (= NR4) NR5C (= NR4) NR4R5, -O (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -S (O) o, 1,2- (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4C (= NR5) NR4R5, -C (= NR4) NR4R5, -C (= NR4) NR4C (O) R6, -NR4SO2R6, -NR4C (O) R6, -NR4C (= O ) OR6, -C (O) NR4R5, - (CR6R7) vC (O) NR4R5, -SO2NR4R5, -Heteroaryl-NR4R5, -Heterocyclyl-NR4R5, -Heteroaryl-N (R4) C (= NR5) NR4R5, -Heterocyclyl- N (R4) C (= NR5) NR4R5, -N (R4) -Heteroaryl-NR4R5, -N (R4) -Heterocyclyl-NR4R5, - (CR6R7) vHeteroaryl-NR4R5, - (CR6R7) vHeterocyclyl-NR4R5, - (CR6R7 ) vHeteroaryl-N (R4) C (= NR5) NR4R5, - (CR6R7) vHeterocyclyl-N (R4) C (= NR5) NR4R5, - (CR6R7) vHeteroaryl, - (CR6R7) vHeterocyclyl, -O-Heteroaryl, -O -Heterocyclyl, -NR4 (CR6R7) vHeteroaryl, -NR4 (CR6R7) vHeterocyclyl, -O (CR6R7) vHeteroaryl, -O (CR6R7) vHeterocyclyl, -NR4 (CR 6R7) vNR5-Heteroaryl, -NR4 (CR6R7) vNR5-Heterocyclyl, -O (CR6R7) vNR5-Heteroaryl, -O (CR6R7) vNR5-Heterocyclyl, -O (CR6R7) vO-Heterocyclyl, -NR4R5, - + Q CR6R7) vNR4R5R9 + Q-, -NR4 (CR6R7) vNR4R5R9 + Q-, -NR4R9 + (CR6R7) vNR4R5R9 + Q-2, - (CR6R7) v (T) + Q-, and -O (CR6R7) vNR4R5R9 ; on what: each T is independently pyridine — 1 — ila, pyrimidin — 1 — ila, or thiazol-3-yl; Q is a pharmaceutically acceptable counterion; and each v is independently 1,2, 3, or 4; or Y taken together with the carbon atom to which it is attached forms an optionally substituted spiro-carbocycle or an optionally substituted spiro-heterocycle; or two Y taken together with the carbon atoms to which they are attached form an optionally substituted carbocycle or an optionally substituted heterocycle; each of R6 and R7 is independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, optionally substituted C1-C6 alkyl, optionally substituted alkoxyalkyl, optionally substituted hydroxyalkyl, optionally substituted C3-C6 cycloalkyl, -NR4C (O) OR5 , -NR4C (O) NR5, -C (O) OR5, -C (N = R5) NR4R5, -OH, -OR10, -SR10, -NR4R5, -NR4C (O) R5, -C (O) NR4R5, -NR4SO2R5, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; or R6 and R7 taken together form an oxo, oxime, or an optionally substituted carbocycle or a heterocycle optionally substituted with the carbon to which they are attached; and each R9 is independently an optionally substituted C1-C6 alkyl.
[0071] [0071] In some embodiments of a Formula I or Formula Ia compound, at least one Y is selected from the group consisting of fluorine, chlorine, optionally substituted C1-C6 alkyl, = O, -OH, -OR10, -NR4R5, - (CR6R7) vNR4R5, - (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4R5 (CR6R7) vR6, -NR4R5 (CR6R7) vHeterocyclyl-C (= NR5) NR4R5, -NR4 (CR6R7) NR4R NR4 (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4 (CR6R7) vNR4R5, -O (CR6R7) vNR4R5, -N (R4) C (O) (CR6R7) vNR4R5, - (CR6R7) ON (R4) C (R4) C) (CR6R7) vNR4R5, -C (O) NR4 (CR6R7) vNR4R5, -S (O) o, i, 2NR4 (CR6R7) vNR4R5, -NR5C (O) NR4 (CR6R7) vNR4R5, -OC (O) NR4 (CR6R7 ) vNR4R5, -NR5C (= NR7) NR4 (CR6R7) vNR4R5, -N (R4) C (= NR5) R6, - (CR6R7) vN (R4) C (= NR5) R6, -NR4 (CR6R7) vN (R4 ) C (= NR5) R6, -O (CR6R7) vN (R4) C (= NR5) R6, - (CR6R7) vC (= NR5) NR4R5, -NR4 (CR6R7) vC (= NR5) NR4R5, -O ( CR6R7) vC (= NR5) NR4R5, - (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4 (CR6R7) vN (R4) C (= NR5) NR4R5, -O (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4C (= NR5) NR4C (= NR5) NR4R5, - (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4 (C R6R7) vC (= NR4) NR5C (= NR4 ) NR4R5, -O (CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4C (= NR5) NR4R5 , -C (= NR4) NR4R5, -C (= NR4) NR4C (O) R6, -NR4SO2R6, -NR4C (O) R6, -NR4C (= O) OR6, -C (O) NR4R5, - (CR6R7) vC (O) NR4R5, -Heteroaryl-NR4R5, -Heterocyclyl-NR4R5, -Heteroaryl-N (R4) C (= NR5) NR4R5, -Heterocyclyl-N (R4) C (= NR5) NR4R5, -N (R4) - Heteroaryl-NR4R5, -N (R4) -Heterocyclyl-NR4R5, - (CR6R7) vHeteroaryl-NR4R5, - (CR6R7) vHeterocyclyl-NR4R5, - (CR6R7) vHeteroaryl-N (R4) C (= NR5) CR4R7, - (CR4R7) - ) vHeterocyclyl-N (R4) C (= NR5) NR4R5, - (CR6R7) vHeteroaryl, - (CR6R7) vHeterocyclyl, -O-Heteroaryl, -O-Heterocyclyl, -NR4 (CR6R7) vHeteroaryl, -NR4 (CR6R7) vHeter -O (CR6R7) vHeteroaryl, -O (CR6R7) vHeterocyclyl, and -O (CR6R7) vO-Heterocyclyl. In certain embodiments, at least one Y is selected from the group consisting of fluorine, optionally substituted C1-C6 alkyl, -OH, -NR4R5, - (CR6R7) vNR4R5, - (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4R5 (CR6R7 ) vR6, -NR4R5 (CR6R7) vHeterocyclyl-C (= NR5) NR4R5, -NR4 (CR6R7) vNR4C (= NR4) NR4R5, -NR4 (CR6R7) vNR4R5 (CR6R7) vNR4R5, -NR4 (CR6R7) CR6R7) vNR4R5, -C (O) NR4 (CR6R7) vNR4R5, -NR5C (O) NR4 (CR6R7) vNR4R5, -NR5C (= NR7) NR4 (CR6R7) vNR4R5, -N (R4) C (= NR5) R6, - (CR6R7) vN (R4) C (= NR5) R6, -NR4 (CR6R7) vN (R4) C (= NR5) R6, - (CR6R7) vC (= NR5) NR4R5, -NR4 (CR6R7) vC (= NR5) NR4R5, - (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4 (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4C (= NR5) NR4C (= NR5) NR4R5, - ( CR6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4 (C R6R7) vC (= NR4) NR5C (= NR4) NR4R5, -NR4C (= NR5) NR4R5, -C (= NR4) NR4R5, -C (= NR4) NR4C (O) R6, -NR4C (O) R6, - (CR6R7) vC (O) NR4R5, -Heterocyclyl-NR4R5, -Heterocyclyl-N (R4) C (= NR5) NR4R5, -N (R4 ) -Heterocyclyl-NR4R5, - (CR6R7) vHeterocyclyl-NR4R5, - (CR6R7) vHeterocyclyl-N (R4) C (= NR5) NR4R5, - (CR6R7) vHeterocyclyl, and -NR4 (CR6R7) vHeterocyclic. In other embodiments, at least one Y is selected from the group consisting of -Heteroaryl-NR4R5, -Heterocyclyl-NR4R5, -Heteroaryl-N (R4) C (= NR5) NR4R5, -Heterocyclyl-N (R4) C (= NR5 ) NR4R5, -N (R4) -Heteroaryl-NR4R5, -N (R4) -Heterocyclyl-NR4R5, -Heteroaryl-C (= NR5) NR4R5, -Heterocyclyl-C (= NR5) NR4R5, - (CR6R7) vHeteroaril-NR4R5 , - (CR6R7) vHeterocyclyl-NR4R5, - (CR6R7) vHeteroaryl-N (R4) C (= NR5) NR4R5, and - (CR6R7) vHeterocyclyl-N (R4) C (= NR5) NR4R5. In the specific embodiments, at least one Y is 2- (NR4R5) -pyridyl, 2- (NR4R5) -pyrimidinyl, 2- (NR4R5) -thiazolyl, 2- (NR4R5) -imidazolyl, 3- (NR4R5) -pyrazolyl, 3 - (R4R5N) -isothiazolyl, 2- (R4R5N) -oxazolyl, piperidine, pyrrolidine, 4-amino-piperidinyl, 3-amino-pyrrolidinyl, piperazine, or 4-carboxyidoyl-piperazinyl. In preferred embodiments, at least one Y is selected from the group consisting of -NR4R5, -NR4C (= NR5) NR4R5, -C (= NR4) NR4R5, -N (R4) C (= NR5) R6, - (CR6R7) vNR4R5, - (CR6R7) vN (R4) C (= NR5) NR4R5, -NR4 (CR6R7) vNR4R5, -NR4 (CR6R7) vOR10, - (CR6R7) vNR4 (CR6R7) vNR4R5, NR5C (= NR5) NR4 vNR4R5, -NR4 (CR6R7) vN (R4) (= NR5) NR4R5, -NR5C (O) CR6 (NR4R5) (CR6R7) vNR4R5, - (CR6R7) vC (= NR5) NR4R5, - (CR6R7) vN (R4) C (O) (CR6R7) vNR4R5, -C (= NR4) NR4C (O) R6, -NR4 (CR6R7) vHeteroaryl, and -O (CR6R7) vNR4R5.
[0072] [0072] In some modalities, p is 0, 1, 2, 3, or 4. In certain modalities, p is 1 or 2. In some modalities, p is 2. In other modalities, p is 1.
[0073] [0073] In some embodiments of a Formula I or Formula Ia compound, each of R4 and R5 is independently selected from the group consisting of hydrogen, -OH, C1-C6 optionally substituted alkyl, optionally substituted alkoxyalkyl, optionally substituted hydroxyalkyl, and optionally substituted heterocyclyl. In preferred embodiments, each of R4 and R5 is independently hydrogen or optionally substituted C1-C6 alkyl.
[0074] [0074] In some embodiments of a Formula I or Formula Ia compound, each of R6 and R7 is independently selected from the group consisting of hydrogen, optionally substituted C1-C6 alkyl, -OH, -NR4R5, and optionally substituted heterocyclyl, or R6 and R7 taken together form a heterocycle optionally substituted with the carbon to which they are attached. In preferred embodiments, each of R6 and R7 is independently hydrogen, fluorine, or optionally substituted C1-C6 alkyl.
[0075] [0075] In some embodiments of a Formula I or Formula Ia compound, R6 and R7 are independently selected from the group consisting of hydrogen, optionally substituted C1-C6 alkyl, -OH, -NR4R5, and optionally substituted heterocyclyl, or R6 and R7 taken together form a heterocycle optionally substituted with the carbon to which they are attached. In preferred embodiments, R6 and R7 are independently hydrogen, fluorine, or optionally substituted C1-C6 alkyl. In some modalities,
[0076] [0076] In some modalities of a compound of Formula I or Formula Ia, in each Y the inclusion of non-hydrogen atoms is defined. For example, in some embodiments, each Y comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 24, 28, 32, 36, 40, 50 or 60 non-hydrogen atoms. In some embodiments each Y comprises less than 50, 40, 36, 32, 28, 24, 20, 18, 16, 14, 12, 10, 9, 8, 7, 6, 5, 4, 3 or 2 atoms not hydrogen. In some embodiments, each Y is independently a group comprising 1 to 50 non-hydrogen atoms. In some embodiments, non-hydrogen atoms are any atoms that are not a hydrogen atom. In some embodiments, non-hydrogen atoms are atoms generally found in organic molecules. In some embodiments, the non-hydrogen atoms are atoms selected from the group consisting of C, N, O, S and P. In some embodiments, each Y is independently a group comprising 1 to 50 non-hydrogen atoms selected from the group consisting of in C, N, O, S, and P.
[0077] [0077] In some modalities of a compound of Formula I or Formula Ia, each Y is defined by its molecular formula. For example, in some modalities, each Y has the formula CwHxNyOz; where each w is independently from 0 to 30; each x is independently from 1 to 69; each y is independently from 1 to 8; and each z is independently from 0 to 10. In some embodiments, each Y has the formula CwHxNyOz; where each w is independently from 0 to 10; each x is independently from 1 to 25; each y is independently from 1 to 4; and each z is independently from 0 to 3. In some embodiments, each y is 2.
[0078] [0078] In some embodiments of a Formula I or Formula Ia compound, each Y is defined by its molecular weight. In some embodiments, each Y has a molecular weight of less than 500, 450, 400, 350, 300, 250, 200, 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 90, 80, 75, 70 or 50 daltons, for example. In some embodiments, each Y has a molecular weight of less than 200 daltons. In some embodiments, each Y has a molecular weight of less than 150 daltons. In some embodiments, each Y has a molecular weight between 30 and 280 daltons.
[0079] [0079] In some embodiments of a Formula I or Formula Ia compound, each Y is defined by the number of basic nitrogen atoms it comprises. For example, each Y can comprise 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or 0 basic nitrogen atoms. In some embodiments, each Y comprises 1 to 6 basic nitrogen atoms. In some embodiments, each Y comprises 1, 2, or 3 basic nitrogen atoms. In some embodiments, each Y comprises 2 basic nitrogen atoms. In some embodiments, at least one Y comprises 1 to 6 basic nitrogen atoms. In some embodiments, at least one Y comprises 1, 2, or 3 basic nitrogen atoms. In some embodiments, at least one Y comprises 2 basic nitrogen atoms. A basic nitrogen atom is a nitrogen atom that can be at least partially protonated in a substantially neutral aqueous buffer. For example, a basic nitrogen atom can be a nitrogen atom of an amine group or a nitrogen atom in a functional group such as an alkyl amine, a cycloalkyl amine, a heterocycloalkyl group, a heteroaryl group comprising a nitrogen , an amidine or a guanidine.
[0080] [0080] In some embodiments of a Formula I or Formula Ia compound, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is 1,4-cyclohexyl; and Y is a group comprising 2 basic nitrogen atoms. In some embodiments, the basic nitrogen atoms are each, an atom within an amine group, and amidine group, a guanidine group, an alkyl heterocycle group, and heteroaryl group, or an amino alkyl group. In some embodiments, Y comprises two groups of amine. In some embodiments, Y comprises two groups of guanidine. In some embodiments, Y comprises an amine group and a guanidine group.
[0081] [0081] In some embodiments of a compound of Formula I or Formula Ia, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is 1,4-cyclohexyl; and Y has the formula CwHxNyOz; where w is 0 to 10; x is 1 to 25; y is 1 to 4; and z is 0 to 3. In some modalities, y is 2. In some modalities, y is 4.
[0082] [0082] In some embodiments of a Formula I or Formula Ia compound, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is 1,4-cyclohexyl; and Y has a molecular weight between 30 and 280 daltons and Y comprises at least 1 basic nitrogen atom. In some embodiments, Y has a molecular weight between 30 and 280 daltons and Y comprises at least 2 basic nitrogen atoms. In some embodiments, Y is an alkyl group comprising 2 amino groups, and Y has a molecular weight between 30 and 280 daltons.
[0083] [0083] In some embodiments of a compound of Formula I or Formula Ia, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is 1,4-cyclohexyl; and Y is -NR4 (CR6R7) vNR4R5; and v is 2. In some embodiments, each of R6 and R7 is independently selected from the group consisting of H, methyl, or OH. In some embodiments, each of R6 and R7 is independently H or methyl. In some embodiments, each of R6 and R7 is H. In some embodiments, each of R4 is H. In some embodiments, R5 is selected from H, optionally substituted C1-C6 alkyl, and optionally substituted C3-C6 cycloalkyl. In some embodiments, R5 is selected from H, optionally substituted C1-C6 alkyl, and optionally substituted C3-C6 cycloalkyl. In some embodiments, R5 is selected from the group consisting of methyl, ethyl, propyl, isopropyl and H. In some embodiments, each of R4, R6, and R7 is H, and R5 is optionally selected from H, C1-C6 alkyl substituted and optionally substituted C3-C6 cycloalkyl. In some embodiments, each R4 is independently H or C1-C3 optionally substituted alkyl; each of R6 and R7 are H; and R5 is selected from H, C1-C6 optionally substituted alkyl and C3-C6 cycloalkyl optionally substituted. In some embodiments, R5 is a guanidine group. In some embodiments, R5 is an amidine group. In some embodiments CycA is trans-1,4-cyclohexyl.
[0084] [0084] In some embodiments of a Formula I or Formula Ia compound, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is 1,4-cyclohexyl; and Y is -NR4R5. In some embodiments, R4 and R5 are each selected from the group consisting of H, guanidine, amidine, optionally substituted alkyl and heterocycloalkyl. In some embodiments, R4 and R5 are each H. In some embodiments, R5 is an amidine group. In some embodiments, R5 is a guanidine group.
[0085] [0085] In some embodiments of a compound of Formula I or Formula Ia, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is 1,4-cyclohexyl; and Y is - (CR6R7) vNR4R5; and v is 1 or 2. In some embodiments, R4 and R5 are each selected from the group consisting of H, guanidine, amidine, optionally substituted alkyl and heterocycloalkyl. In some embodiments, v is 1; R6 and R7 are each H or methyl; and R4 and R5 are each independently H, optionally substituted C1-C6 alkyl or optionally substituted C3-C6 cycloalkyl. In some embodiments, v is 1; R6 and R7 are each H; and R4 and R5 are each independently H, optionally substituted C1-C6 alkyl or optionally substituted C3-C6 cycloalkyl. In some embodiments, v is 1; R6 and R7 are each of H; and R4 and R5 are each H, C1-C6 alkyl or C3-C6 cycloalkyl. In some embodiments, v is 1; R6 and R7 are each H or methyl; and R4 and R5 are each H.
[0086] [0086] In some embodiments of a compound of Formula I or Formula Ia, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is 1,4-cyclohexyl; and Y is - (CR6R7) vNR4C (= NR4) NR4R5; and v is 1 or 2. In some embodiments, R4 and R5 are each selected from the group consisting of H, optionally substituted C1-C6 alkyl or optionally substituted C3-C6 cycloalkyl. In some embodiments, v is 1; R6 and R7 are each H or methyl; and R4 and R5 are each independently H, optionally substituted C1-C6 alkyl or optionally substituted C3-C6 cycloalkyl. In some embodiments, v is 1; R6 and R7 are each H; and R4 and R5 are each independently H, optionally substituted C1-C6 alkyl or optionally substituted C3-C6 cycloalkyl. In some embodiments, v is 1; R6 and R7 are each H; and R4 and R5 are each H, C1-C6 alkyl, or C3-C6 cycloalkyl. In some embodiments, v is 1; R6 and R7 are each H or methyl; and R4 and R5 are each H.
[0087] [0087] In some embodiments of a Formula I or Formula Ia compound, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is 1,4-cyclohexyl; and Y is -NR4 (CR6R7) vNR4C (= NR4) NR4R5; and v is 2. In some embodiments, R4 and R5 are each selected from the group consisting of H, optionally substituted alkyl and heterocycloalkyl. In some embodiments, R6 and R7 are each H or methyl; and R4 and R5 are each independently H, optionally substituted C1-C6 alkyl or optionally substituted C3-C6 cycloalkyl. In some modalities; R6 and R7 are each H; and R4 and R5 are each independently H, optionally substituted C1-C6 alkyl or optionally substituted C3-C6 cycloalkyl. In some embodiments, R6 and R7 are each H; and R4 and R5 are each H, C1-C6 alkyl or C3-C6 cycloalkyl. In some embodiments, R6 and R7 are each H or methyl; and R4 and R5 are each H.
[0088] [0088] In some embodiments of a Formula I or Formula Ia compound, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is 1,4-cyclohexyl; and Y is - (CR6R7) v; v is 1 or 2; each R7 is H or methyl; and at least one R6 is -C (N = R5) NR4R5. In some embodiments, each of R4 and R5 is selected from H, optionally substituted C1-C6 alkyl and optionally substituted C3-C6 cycloalkyl. In some embodiments, each of R4 and R5 is selected from H, C1-C6 alkyl, and C3-C6 cycloalkyl. In some embodiments, each of R4 and R5 is selected from H and methyl. In some modalities, each of R4 and R5 is H.
[0089] [0089] In some embodiments of a compound of Formula I or Formula Ia, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH, Z is> C = O; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; Y is -NR4 (CR6R7) vNR4R5; and v is 2; each of R6 and R7 is independently selected from the group consisting of H, methyl, or OH; each R4 is H; R5 is selected from H, C1-C6 optionally substituted alkyl and C3-C6 cycloalkyl optionally substituted; and CycA is an optionally substituted 3- to 10-membered non-aromatic carbocycle, in which an optional olefin functionality of the non-aromatic carbocycle is not directly linked to an oxygen, sulfur or nitrogen substituent. In some embodiments, CycA is selected from the group consisting of cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclopentene, cyclohexene, cycloheptene and cyclooctene, in which the olefin functionality Cyclopentene, cyclohexene, cycloheptene and cyclooctene is not directly linked to an oxygen, sulfur or nitrogen substituent. In certain embodiments, CycA is cyclobutane, cyclopentane, cyclohexane or cyclohexene, in which the olefin functionality of cyclohexene is not directly linked to an oxygen, sulfur or nitrogen substituent. In other modalities, CycA is selected from the group consisting of octane bicycles [3.3.0], nonane bicycles [4.3.0], cis-decalines, trans-decalines, bicycles [2.1.1] hexane, bicycles [2.2.1] heptane, bicycles [2.2.2] octane, bicycles [3.2.2] nonane, and bicycles [3.3.2] decane. In some embodiments, CycA is cyclopentane. In preferred embodiments, CycA is cyclohexane. In some embodiments, CycA is cyclohexane covalently linked to one of Y and L; said covalent bonds in the 1,4-trans arrangement.
[0090] [0090] In some embodiments of a Formula I or Formula Ia compound, Ra, Rb, Rc, Rd, and R3 are H; X1 is OH; X2 when present is OH; n is 0; m is 1; p is 1; M and L are each a link; R1 and R2 are each H; CycA is cyclohexane, cyclopentane or cyclobutane; Y is -NR4 (CR6R7) vNR4R5; and v is 2; each of R6 and R7 is independently selected from the group consisting of H, methyl, or OH; each R4 is H; R5 is selected from H, C1-C6 optionally substituted alkyl and C3-C6 cycloalkyl optionally substituted; and Z is selected from the group consisting of> C = O,> C = S, or> SO2. Preparation of Compounds
[0091] [0091] Compounds of Formula I or Formula Ia which inhibit the activity of beta-lactamases, and processes for their preparation are described herein. Also described herein are pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically active metabolites and pharmaceutically acceptable prodrugs of such compounds. Pharmaceutical compositions comprising at least one such compound or a pharmaceutically acceptable salt, a pharmaceutically acceptable solvate, a pharmaceutically active metabolite or a pharmaceutically acceptable prodrug of such a compound, and a pharmaceutically acceptable excipient are also provided.
[0092] [0092] Formula I or Formula Ia compounds can be synthesized using conventional synthetic reactions known to those of skill in the art or using methods known in the art. The reactions can be used in a linear sequence to provide the compounds or they can be used to synthesize the fragments that are subsequently ligated by methods known in the art.
[0093] [0093] The starting material used for the synthesis of the compounds described herein can be synthesized or obtained from commercial sources, such as, but not limited to, Aldrich Chemical Co. (Milwaukee, Wisconsin), Bachem (Torrance, California ), or Sigma Chemical Co. (St. Louis, Mo.). The compounds described herein, and other related compounds having different substituents can be synthesized using techniques and materials known to those of skill in the art, such as described, for example, in March, Advanced Organic Chemistry 4th Ed., (Wiley 1992); Carey and Sundberg, Advanced Organic Chemistry 4th Ed., Vols. A and B (Plenum 2000, 2001); Green and Wuts, Protective Groups in Organic Synthesis 3rd Ed., (Wiley 1999); Fieser and Fieser’s Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd’s Chemistry of Carbon Compounds, Volumes 1-5 and Supplemental (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991); and Larock’s Comprehensive Organic Transformations (VCH Publishers Inc., 1989). (All of which are incorporated herein by reference in their entirety). Other methods for the synthesis of the compounds described herein can be found in International Patent Publication No. WO 01/01982901, Arnold et al. Bioorganic & Medicinal Chemistry Letters 10 (2000) 2167-2170; Burchat et al. Bioorganic & Medicinal Chemistry Letters 12 (2002) 1687-1690. The general methods for preparing the compound as disclosed herein, can be derived from the reactions known in the field, and the reactions can be modified through the use of appropriate reagents and conditions, as would be recognized by the skilled person, with respect to the introduction of the various components found in the formulas as provided here.
[0094] [0094] The reaction products can be isolated and purified, if desired, using conventional techniques, including, but not limited to, filtration, distillation, crystallization, chromatography and the like. Such materials can be characterized using conventional means, including physical constants and spectral data.
[0095] [0095] The compounds described herein can be prepared as an isolated isomer or a mixture of isomers. Additional Forms of the Compounds Disclosed Here Isomers
[0096] [0096] In some embodiments, due to the oxophilic nature of the boron atom, the compounds described here can convert or exist in equilibrium with alternative forms, particularly in water-containing media (aqueous solution, plasma, etc.). Consequently, the compounds described here can exist in a balance between the "closed" cyclic form shown in Formula I and the "open" acyclic form shown in Figure Ia. In addition to the compounds described here, they can associate in intramolecular dimers, trimers and related combinations .
[0097] [0097] In addition, in some embodiments, the compounds described here exist as geometric isomers. In some embodiments, the compounds described herein have one or more double bonds. The compounds presented herein include all cis, trans, syn, anti, entgegen (E), and zusammen (Z) isomers, as well as their corresponding mixtures. In some situations, the compounds exist as tautomers. The compounds described herein include all possible tautomers within the formulas described herein. In some situations, the compounds described herein have one or more chiral centers and each center exists in the R configuration or the S configuration. The compounds described herein include all diastereomeric, enantiomeric and epimeric forms, as well as their corresponding mixtures. In the additional embodiments of the compounds and methods provided herein, mixtures of enantiomers and / or diastereoisomers, resulting from a single stage of preparation, combination or interconversion, are useful for the applications described here. In some embodiments, the compounds described here are prepared as their individual stereoisomers by reacting a racemic mixture of the compound with an optically active resolving agent to form a pair of diastereoisomeric compounds, separating the diastereomers and recovering the optically pure enantiomers. In certain embodiments, dissociable complexes are preferred (for example, crystalline diastereomeric salts). In certain embodiments, diastereomers have distinct physical properties (for example, melting points, boiling points, solubilities, reactivity, etc.) and are separated taking advantage of these dissimilarities. In some embodiments, the diastereomers are separated by chiral chromatography, or preferably, by separation / resolution techniques based on differences in solubility. In some embodiments, the optically pure enantiomer is then recovered, along with the resolving agent, by any practical means that do not result in racemization. Labeled compounds
[0098] [0098] In some embodiments, the compounds described here exist in their isotopically labeled forms. In some embodiments, the methods described herein include methods of treating disease by administering such isotopically labeled compounds. In some embodiments, the methods disclosed herein include methods of treating disease by administering such isotopically labeled compounds as pharmaceutical compositions. Thus, in some embodiments, the compounds described herein include isotopically labeled compounds, which are identical to those recited here, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as 2H, 3H, 13C, 14C, 15N, 180, 17O, 31P, 32P, 35S, 18F, and 36Cl, respectively. The compounds described herein, and their metabolites, salts, esters, prodrugs, solvates, hydrates or pharmaceutically acceptable derivatives containing the aforementioned isotopes and / or other isotopes of other atoms, are within the scope of this invention. Certain isotopically labeled compounds, for example, those whose radioactive isotopes such as 3H and 14C are incorporated, are useful in medications and / or substrate tissue distribution assays. Tritiated isotopes, i.e., 3H and carbon-14, i.e., 14C, are particularly preferred for their ease of preparation and detection capability. In addition, substitution with heavy isotopes such as deuterium, i.e., 2H, produces certain therapeutic advantages resulting from increased metabolic stability, for example, increased in vivo half-life or reduced dosage requirements. In some embodiments, isotopically labeled compounds, their salts, ester, prodrug, solvate, hydrate or pharmaceutically acceptable derivative, are prepared by any suitable method.
[0099] [0099] In some embodiments, the compounds described herein are labeled by other means, including, but not limited to, the use of chromophores or fluorescent components, bioluminescent labels or chemiluminescent labels. Pharmaceutically acceptable salts
[0100] In some embodiments, the compounds described herein exist as their pharmaceutically acceptable salts. In some embodiments, the methods disclosed herein include methods of treating disease by administering such pharmaceutically acceptable salts. In some embodiments, the methods disclosed herein include methods of treating disease by administering such pharmaceutically acceptable salts as pharmaceutical compositions.
[0101] [00101] In some embodiments, the compounds described herein have acidic or basic groups and therefore react with any of a number of inorganic or organic bases and inorganic and organic acids, to form a pharmaceutically acceptable salt. In certain embodiments, these salts are prepared in situ during the final isolation and purification of the compounds of the invention, or by separately reacting a purified compound in its free form with a suitable acid or base, and isolating the salt so formed.
[0102] [00102] Examples of pharmaceutically acceptable salts include salts prepared by reacting the compounds described herein with a mineral, organic acid or inorganic base, such salts including, acetate, acrylate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, bisulfite, bromide, butyrate, butin-1,4-dioate, camphorate, camphorsulfonate, caproate, caprylate, chlorobenzoate, chloride, citrate, cyclopentanopropionate, decanoate, digluconate, dihydrogen phosphate, dinitrobenzoate, dodecyl sulfate, ethanesulfonate, gluten, formia glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hexino-1,6-dioate, hydroxybenzoate, γ-hydroxybutyrate, hydrochloride, hydrobromide, iodhydride, 2-hydroxyethanesulfonate, iodide, isobutyrate, lactate, maleate, malonate, methane, malonate, methane methanesulfonate, methoxybenzoate, methylbenzoate, monohydrogen phosphate, 1-naphthalenesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, palmoate, pectin ato, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, pyrosulfate, pyrophosphate, propiolate, phthalate, phenylacetate, phenylbutyrate, propanesulfonate, salicylate, succinate, sulphate, sulphite, succinate, suberate, sebacate, tartarate, sebacate, tartarate tosylate, undeconate and xylenesulfonate.
[0103] [00103] Furthermore, the compounds described herein can be prepared as pharmaceutically acceptable salts formed by reacting the free base form of the compound with a pharmaceutically acceptable inorganic or organic acid, including, but not limited to, inorganic acids such as hydrochloric acid hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, and the like; and organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanopropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, p-toluenesulfonic acid, tartaric acid, trifluoroacetic acid , citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, aryl sulfonic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanesulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo- [2.2.2] oct-2-ene-1-carboxylic acid, glucoeptonic acid, 4,4'-methylenebis- (3-hydroxy-2-ene-1-carboxylic acid), acid 3-phenylpropionic, trimethyl-cetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxinaftoic acid, salicylic acid, stearic acid and muconic acid. In some embodiments, other acids, such as oxalic, although in isolation not pharmaceutically acceptable, are employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts.
[0104] [00104] In some embodiments, those compounds described herein that comprise a free acid group react with a suitable base such as hydroxide, carbonate, bicarbonate, sulfate, of a pharmaceutically acceptable metal cation, with ammonia, or with a primary organic amine, secondary, tertiary or pharmaceutically acceptable quaternary. Representative salts include alkaline or alkaline earth salts, such as lithium, sodium, potassium, calcium, magnesium and aluminum salts and the like. Illustrative examples of bases include sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, N + (C1-4 alkyl) 4, and the like.
[0105] Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylene diamine, ethanolamine, diethanolamine, piperazine and more. It should be understood that the compounds described herein include the quaternization of any groups containing basic nitrogen that they contain. It should be understood that the compounds described herein also include the quaternization of any boron-containing groups that they contain. Such quaternization can result from the treatment of Lewis acidic boron with a Lewis base to form a complex or a salt thereof. In some embodiments, water-dispersible or oil-soluble products are obtained through such quaternization. Solvates
[0106] [00106] In some embodiments, the compounds described herein exist as solvates. The invention provides methods of treating disease by administering such solvates. The invention further provides methods of treating disease by administering such solvates as pharmaceutical compositions.
[0107] [00107] Solvates contain stoichiometric or non-stoichiometric amounts of a solvent, and, in some embodiments, are formed during the crystallization process with pharmaceutically acceptable solvents such as water, ethanol, and the like. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is alcohol. The solvates of the compounds described herein can be conveniently prepared or formed during the processes described herein. By way of example only, the hydrates of the compounds described herein can be conveniently prepared by recrystallizing an aqueous / organic solvent mixture, using organic solvents including, but not limited to, dioxane, tetrahydrofuran or methanol. In addition, the compounds provided herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the compounds and methods provided herein. Polymorphs
[0108] [00108] In some embodiments, the compounds described herein exist as polymorphs. The invention provides methods of treating disease by administering such polymorphs. The invention further provides methods of treating disease by administering such polymorphs as pharmaceutical compositions.
[0109] [00109] Thus, the compounds described herein include all of their crystalline forms known as polymorphs. Polymorphs include the different crystalline packaging arrangements of the same elemental composition as a compound. In certain cases, polymorphs have different X-ray diffraction patterns, infrared spectra, melting points, density, resistance, crystal shape, optical and electrical properties, stability and solubility. In certain cases, several factors such as the recrystallization solvent, crystallization rate and storage temperature cause a unique crystal form to dominate. Prodrugs
[0110] [00110] In some embodiments, the compounds described herein exist in the form of a prodrug. The invention provides methods of treating disease by administering such prodrugs. The invention further provides methods of treating disease by administering such prodrugs as pharmaceutical compositions.
[0111] [00111] Prodrugs are generally drug precursors that, after administration to an individual and subsequent absorption, are converted into an active principle, or a more active species through some process, such as conversion by a metabolic route. Some prodrugs have a chemical group present in the prodrug that makes it less active and / or confers solubility or some other property to the drug. As soon as the chemical group has been cleaved and / or modified from the prodrug, the active drug is generated. Prodrugs are often useful because, in some situations, they are easier to administer than the original drug. They are, for example, bioavailable by oral administration, while of origin they are not. In certain cases, the prodrug also improved the solubility in pharmaceutical compositions over the original drug. An example, without limitation, of a prodrug, would be a compound as described herein which is administered as an ester (the "prodrug") to facilitate transmission across a cell membrane where water solubility is detrimental to mobility, but which is then metabolically hydrolyzed to carboxylic acid, the active entity, as well as inside the cell where water solubility is beneficial. Another example of a prodrug can be a short peptide (polyamino acid) attached to an acid group, where the peptide is metabolized to reveal the active component. (See, for example, Bundgaard, "Design and Application of Prodrugs" in A Textbook of Drug Design and Development, Krosgaard-Larsen and Bundgaard, Ed., 1991, Chapter 5, 113-191, which is incorporated herein by reference).
[0112] [00112] In certain embodiments, prodrugs are designed as reversible drug derivatives, for use as modifiers to improve drug transport to specific tissues at the site. The design of prodrugs to date has been to increase the effective water solubility of the therapeutic compound for targeting in regions where water is the main solvent.
[0113] [00113] Additionally, prodrug derivatives of the compounds described herein that can be prepared by the methods described herein are otherwise known in the art (for more details see Saulnier et al., Bioorganic and Medicinal Chemistry Letters, 1994, 4, 1985). By way of example only, appropriate prodrugs can be prepared by reacting a non-derivatized compound with a suitable carbamylating agent, such as, but not limited to, 1,1-acyloxyalkylcarbanochloridate, para-nitrophenyl carbonate, or the like . The prodrug forms of the compounds described herein, wherein the prodrug is metabolized in vivo to produce a derivative as presented herein, are included within the scope of the claims. In fact, some of the compounds described here are prodrugs for another derivative or active compound.
[0114] [00114] In some embodiments, prodrugs include compounds in which an amino acid residue, or a polypeptide chain of two or more (e.g., two, three or four) amino acid residues, is covalently joined through an amide bond or ester to an amino, hydroxy or carboxylic acid free group of compounds of the present invention. Amino acid residues include, but are not limited to, the 20 naturally occurring amino acids and also include 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid, cirtulline , homocysteine, homossein, ornithine and methionine sulfone. In other embodiments, prodrugs include compounds in which a nucleic acid residue, or an oligonucleotide of two or more (e.g., two, three or four) nucleic acid residues, is covalently linked to a compound of the present invention.
[0115] [00115] Pharmaceutically acceptable prodrugs of the compounds described herein include, but are not limited to, esters, carbonates, thiocarbonates, N-acyl derivatives, N-acyloxyalkyl derivatives, quaternary tertiary amine derivatives, N-Mannich bases, Schiff bases, amino acid conjugates, phosphate esters, metal salts and sulfonate esters. Compounds having free amino, starch, hydroxy or carboxylic groups can be converted into prodrugs. For example, carboxyl-free groups can be derivatized as amides or alkyl esters. In certain cases, all of these prodrug components incorporate groups including, but not limited to, ether, amine and carboxylic acid functionalities.
[0116] [00116] Hydroxy prodrugs include esters, such as, but not limited to, acyloxyalkyl esters (e.g., acyloxymethyl, acyloxyethyl), alkoxycarbonyloxyalkyl esters, alkyl esters, aryl esters, phosphate esters, sulfon ester esters and disulfide-containing esters; ethers, amides, carbamates, hemisuccinates, dimethylaminoacetates and phosphoryloxythyloxycarbonyls, as outlined in Advanced Drug Delivery Reviews 1996, 19, 115.
[0117] [00117] The amine derived prodrugs include, but are not limited to, the following groups and combinations of groups:
[0118] [00118] In certain cases, the sites in any of the aromatic ring components are susceptible to various metabolic reactions, therefore, the incorporation of appropriate substituents into the aromatic ring structures can reduce, minimize or eliminate this metabolic pathway. Metabolites
[0119] [00119] In some embodiments, the compounds of Formula I or Formula Ia are susceptible to various metabolic reactions. Therefore, in some embodiments, the incorporation of appropriate substituents into the structure will reduce, minimize or eliminate a metabolic pathway. In the specific embodiments, the appropriate substituent to decrease or eliminate the susceptibility of an aromatic ring to metabolic reactions is, by way of example only, a halogen, or an alkyl group.
[0120] [00120] In additional or new embodiments, the compounds of Formula I or Formula Ia described herein are metabolized after administration to an organism in need of producing a metabolite which is then used to produce a desired effect, including a desired therapeutic effect. Pharmaceutical Compositions / Formulations
[0121] [00121] In another aspect, a pharmaceutical composition is provided herein comprising a compound of Formula I or Formula Ia as described herein, or its pharmaceutically acceptable salt, polymorph, solvate, prodrug, N-oxide or isomer, and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition further comprises a beta-lactam antibiotic. In certain embodiments, the beta-lactam antibiotic is a penicillin, cephalosporin, carbapenem, monobactam, bridged monobactam, or a combination of these.
[0122] [00122] In some embodiments, the compounds described herein are formulated in pharmaceutical compositions. The pharmaceutical compositions are formulated in a conventional manner using one or more pharmaceutically acceptable inactive ingredients that facilitate the processing of the active compounds in the preparations that can be used pharmaceutically. The appropriate formulation is dependent on the route of administration selected. A summary of the pharmaceutical compositions described herein can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa .: Mack Publishing Company, 1995); Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins1999), incorporated herein by reference for such description.
[0123] [00123] Pharmaceutical compositions which include a compound of Formula I or Formula Ia and at least one inactive ingredient pharmaceutically acceptable carrier are provided herein. In some embodiments, the compounds described herein are administered as pharmaceutical compositions in which a compound of Formula I or Formula Ia is mixed with other active ingredients, as in combination therapy. In other embodiments, the pharmaceutical compositions include other medicinal or pharmaceutical agents, carriers, adjuvants, preservatives, stabilizers, humectants or emulsifiers, solution promoters, salts for regulating osmotic pressure and / or buffers. In yet other embodiments, the pharmaceutical compositions include other therapeutically valuable substances.
[0124] [00124] A pharmaceutical composition, as used herein, refers to a mixture of a Formula I or Formula Ia compound with other chemical components (i.e. pharmaceutically acceptable inactive ingredients), such as carriers, excipients, binders, agents fillers, suspending agents, flavoring agents, sweetening agents, disintegrating agents, dispersing agents, surfactants, lubricants, dyes, diluents, solubilizers, wetting agents, plasticizers, stabilizers, penetration enhancers, wetting agents, antifoaming agents, antioxidants , preservatives, or one or more of its combinations. The pharmaceutical composition facilitates the administration of the compound in an organism. In practicing the methods of treatment or use provided herein, the therapeutically effective amounts of the compounds described herein are administered in a pharmaceutical composition to a mammal having a disease, disorder or condition to be treated. In some embodiments, the mammal is a human being. A therapeutically effective amount can vary widely depending on the severity of the disease, the age and relative health of the individual, the potency of the compound used and other factors. The compounds can be used alone or in combination with one or more therapeutic agents as components of mixtures.
[0125] [00125] The pharmaceutical formulations described herein are administered to an individual through the appropriate administration routes, including, but not limited to, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular), intranasal, buccal, topical administration routes , rectal or transdermal. The pharmaceutical formulations described herein include, but are not limited to, aqueous liquid dispersions, liquids, gels, syrups, elixirs, slurries, suspensions, self-emulsifying dispersions, solid solutions, liposome dispersions, aerosols, solid oral dosage forms, powders , immediate-release formulations, controlled-release formulations, fast-dissolving formulations, tablets, capsules, pills, powders, pills, effervescent formulations, lyophilized formulations, delayed-release formulations, prolonged-release formulations, pulsatile-release formulations, multiparticulate formulations and mixed and immediate release formulations.
[0126] [00126] Pharmaceutical compositions including a compound of Formula I or Formula Ia are prepared in a conventional manner, such as, by way of example only, by means of conventional mixing, dissolving, granulating, drug-making, levigation, emulsification processes , encapsulation, capture or compression.
[0127] [00127] The pharmaceutical compositions will include at least one compound of Formula I or Formula Ia as an active ingredient in the form of free acid or free base, or in a pharmaceutically acceptable salt form. In addition, the pharmaceutical methods and compositions described herein include the use of N-oxides (if appropriate), crystalline forms, amorphous phases, as well as active metabolites of these compounds having the same type of activity. In some embodiments, the compounds described herein exist in unsolvated form or in forms solvated with pharmaceutically acceptable solvents such as water, ethanol, and the like. The solvated forms of the compounds presented here are also considered to be disclosed herein.
[0128] [00128] Pharmaceutical preparations for oral use are obtained by mixing one or more solid excipients with one or more of the compounds described herein, optionally grinding the resulting mixture, and processing the granule mixture, after adding suitable auxiliaries, if desired , to obtain tablets or pill cores. Suitable excipients include, for example, fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations such as, for example, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or others such as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. If desired, disintegrating agents are added, such as cross-linked sodium croscarmellose, polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate. In some embodiments, dyestuffs or pigments are added to tablets or dragee coatings for identification or to characterize the different combinations of doses of active compounds.
[0129] [00129] Pharmaceutical preparations that are administered orally include controlled-release capsules produced from gelatin, as well as soft capsules, sealed capsules produced from gelatin and a plasticizer, such as glycerol or sorbitol. Controlled release capsules contain the active ingredients in admixture with the filler such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds are dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In some embodiments, stabilizers are added.
[0130] [00130] In certain embodiments, delivery systems for pharmaceutical compounds can be used, such as, for example, liposomes and emulsions. In certain embodiments, the compositions provided herein may also include a mucus-adhesive polymer, selected from, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly (methyl methacrylate), polyacrylamide, polycarbophile, acrylic acid / acrylate copolymer of butyl, sodium alginate and dextran. Combination Treatment
[0131] [00131] The compounds according to Formula I or Formula Ia can be used in combination with one or more antibiotics in the treatment of bacterial infections. Such antibiotics can be administered, by a route and in an amount commonly used, therefore, contemporary or sequentially with a compound of Formula I or Ia. When a compound of Formula I or Ia is used simultaneously with one or more antibiotics, a pharmaceutical composition in unit dosage form containing such other drugs and the compound of the present invention is preferable. However, combination therapy can also include therapies in which the compound of Formula I or IA and one or more antibiotics are administered at different overlapping times. It is also contemplated that when used in combination with one or more antibiotics, antibiotics can be used in lower doses than when each is used alone.
[0132] Accordingly, the pharmaceutical compositions of the present invention also include those containing one or more antibiotics, in addition to a compound according to Formula I or Formula Ia. In some embodiments, a pharmaceutical composition comprising a compound of Formula I or Ia further comprises a beta-lactam antibiotic. In certain embodiments, the beta-lactam antibiotic is a penicillin, cephalosporin, carbapenem, monobactam, bridged monobactam, or a combination of these.
[0133] [00133] The above combinations include combinations of a compound of Formula I or Ia not only with one antibiotic, but also with two or more antibiotics. Likewise, compounds of formula I or Ia, in combination with an antibiotic or alone, can be used in combination with other drugs that are used to prevent, treat, control, improve or reduce the risk of bacterial infections or conditions associated with bacterial infections. Such other medications can be administered, via a route and in a commonly used amount, therefore, contemporaneously or sequentially with a compound of Formula I or Ia. When a compound of Formula I or Ia is used concurrently with one or more of other medications, a pharmaceutical composition containing these drugs other than the compound of the present invention is preferable. Accordingly, the pharmaceutical compositions of the present invention also include those that also contain one or more other active ingredients, in addition to a compound of Formula I or Ia. The weight ratio of the compound of Formula I or Ia to the second active ingredient can be varied and will depend on the effective dose of each ingredient. Generally, an effective dose of each will be used.
[0134] [00134] In some embodiments, compounds according to Formula I or Formula Ia are used in combination with one or more antibiotics in the treatment of bacterial infections. In certain embodiments, bacterial infection is an upper or lower respiratory tract infection, an urinary tract infection, an intra-abdominal infection, or a skin infection. In some embodiments, the one or more antibiotics are selected from β-lactam antibiotics. Β-lactam antibiotics include, but are not limited to, penicillins, penems, carbapenems, cephalosporins, cephamycins, monobactams, or combinations thereof. Penicillins include, but are not limited to, amoxicillin, ampicillin, azidocillin, azlocillin, bacampicillin, benzyl benzylpenicillin, benzoxy phenoxymethylpenicillin, benzylpenicillin (G), carbenicillin, carindacillin, clomethyl, cloxacillin, detoxoxyl metampicillin, methicillin, mezlocillin, nafcillin, oxacillin, penamecillin, pheneticillin, phenoxymethylpenicillin (V), piperacillin, pivampicillin, pivmecillin, procaine benzylpenicillin, propicillin, sulbenicillin, temampicin, tembicicillin, temamp. The penalties include, but are not limited to, pharopenemus. The carbapenems include, but are not limited to, biapenem, ertapenem, doripenem, imipenem, meropenem, panipenem. Cephalosporins / cephamycins include, but are not limited to, cefacetril, cefaclor, cefadroxil, cephalexin, cephaloglycine, cephalonium, cephaloridine, cephalotin, cephaphrine, cefatrizine, cephaphrine, cefazefe, cefazefe, cefazefe, cefazefe, , cefepime, cefetamet, cefixime, cefmenoxime, cefmetazole, cefminox, cefodizime, cefonicide, cefoperazone, ceforanide, cefotaxime, cefotetan, cefotiam, cefovecine, cefoxitine, cefmaprime, cefoze, cefoze, cefoze , cefsulodine, ceftaroline fosamila, ceftazidime, cefteram, ceftezol, ceftibuten, ceftiofur, ceftiolene, ceftizoxime, ceftobiprol, ceftriaxone, cefuroxime, cefuzonam, flomoxef, latamoxef, loracarbef. Monobactams include, but are not limited to, aztreonam, carumonam, nocardicin A, tigemonam. Pharmaceutical Composition Administration
[0135] [00135] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ophthalmic, pulmonary, transmucosal, transdermal, vaginal, optic, nasal and topical administration. Furthermore, by way of example only, parenteral release includes intramuscular, subcutaneous, intravenous, intramedullary injections, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic and intranasal injections.
[0136] [00136] In some embodiments, the compounds of Formula I or Formula Ia and their compositions are administered in any suitable manner. The mode of administration can be selected based, for example, on the desirable treatment, whether local or systemic, and on the area to be treated. For example, the compositions can be administered orally, parenterally (e.g., intravenously, subcutaneously, intraperitoneally or intramuscularly), by inhalation, extracorporeal, topical (including transdermally, ophthalmicly, vaginally, rectally, intranasally) or similar.
[0137] [00137] Parenteral administration of the composition, if used, is generally characterized by injection. Injectables can be prepared in conventional forms, as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid before injection, or as emulsions. A more recently revised approach to parenteral administration involves the use of a slow-release or sustained-release system, such that a constant dosage is maintained. Assays for Antibacterial Activity
[0138] [00138] Assays for inhibiting beta-lactamase activity are well known in the art. For example, the ability of a compound to inhibit beta-lactamase activity in a standard enzyme inhibition assay can be used (see, for example, Page, Biochem J, 295: 295-304 (1993)). Beta-lactamases for use in such assays can be purified from bacterial sources or are preferably produced by recombinant DNA techniques, since the genes and cDNA clones that encode many beta-lactamases are known (see, for example, example, Cartwright & Waley, Biochem J 221: 505-12 (1984)).
[0139] [00139] Alternatively, the sensitivity of known or planned bacteria to produce a beta-lactamase for an inhibitor can be determined. Other bacterial inhibition assays include agar disk diffusion and agar dilution (see, for example, Traub & Leonhard, Chemotherapy 43 159-67 (1997)). Thus, a beta-lactamase can be inhibited by contacting the beta-lactamase enzyme with an effective amount of a compound of the invention or by contacting the bacteria that produce beta-lactamase enzymes with an effective amount of such a compound, so that beta-lactamase in bacteria is brought into contact with the inhibitor. Contact can occur in vitro or in vivo. "Contact" means that the beta-lactamase and the inhibitor are brought together so that the inhibitor can bind to the beta-lactamase. The amounts of a compound effective to inhibit a beta-lactamase can be determined empirically, and making such determinations is within the skill of the art. Inhibition includes both reduction and elimination of beta-lactamase activity. Methods
[0140] [00140] The present description also provides methods for inhibiting bacterial growth, for example, by reducing bacterial resistance to a β-lactam antibiotic, such methods comprising contacting a bacterial cell culture, or a cell culture, tissue or organism infected by bacteria, with a beta-lactamase inhibitor described here. Preferably, the bacteria to be inhibited by administration of a Formula I or Ia beta-lactamase inhibitor are bacteria that are resistant to beta-lactamase antibiotics. The term "resistant" is well understood by those of practical skill in the art (see, for example, Payne et al., Antimicrobial Agents and Chemotherapy 38 767-772 (1994), Hanaki et al., Antimicrobial Agents and Chemotherapy 30 1120- 1126 (1995)).
[0141] [00141] These methods are useful for inhibiting the growth of bacteria in a variety of contexts. In certain embodiments, a compound of Formula I or Ia is administered to an experimental cell culture in vitro to prevent the growth of bacteria resistant to beta-lactams. In certain other embodiments, a compound of Formula I or Ia is administered to a mammal, including a human, to prevent the growth of bacteria resistant to beta-lactam in vivo. The method according to this embodiment comprises administering a therapeutically effective amount of a beta-lactamase inhibitor over a therapeutically effective period of time to a mammal, including a human. Preferably, the beta-lactamase inhibitor is administered in the form of a pharmaceutical composition as described above. In some embodiments, a beta-lactam antibiotic is co-administered with the beta-lactamase inhibitor as described above.
[0142] [00142] In another aspect provided herein are methods for treating a bacterial infection, the method of which comprises administering to a subject a pharmaceutical composition comprising a compound of Formula I or Formula Ia, or a pharmaceutically acceptable salt thereof, and a pharmaceutically excipient acceptable. In some embodiments, methods of treating a bacterial infection in an individual comprise administering to the individual a pharmaceutical composition as described herein, optionally in combination with a beta-lactam antibiotic. In some embodiments, bacterial infection is an upper or lower respiratory tract infection, an urinary tract infection, an intra-abdominal infection, or a skin infection.
[0143] [00143] In some embodiments, the infection that is treated or prevented comprises a bacterium that includes Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas acidovorans, Pseudomonas alkaligenes, Pseudomonas putida, Stenotrophomonas maltophilia, Burkholderia cepacia, Aeromonas hydrophilia, Escherichia colophila, typhimurium, Salmonella typhi, Salmonella paratyphi, Salmonella enteritidis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescensis, Proteella, Morganella tisalensis, Proteella Providencia rettgeri, Providencia stuartii, Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter haemolyticus, Yersinia enterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Yersinia intermedia, Bordetella pertussis, Bordetella parapertussis, Bordetella, Bordetella mophilus influenzae, Haemophilus parainfluenzae, Haemophilus haemolyticus, Haemophilus parahaemolyticus, Haemophilus ducreyi, Pasteurella multocida, Pasteurella haemolytica, Branhamella catarrhalis, Helicobella pylori, Campylobyacter, Vibrant, , Neisseria gonorrhoeae, Neisseria meningitidis, Kingella, Moraxella, Gardnerella vaginalis, Bacteroides fragilis, Bacteroides distasonis, Bacteroides 3452A homology group, Bacteroides vulgatus, Bacteroides ovalus, Bacteroides thetaiotaomicron, Bacteroides bacteroides bactericides , Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium leprae, Corynebacterium diphtheriae, Corynebacterium ulcerans, Streptococcus pneumoniae, Streptococcus agalactiae, Streptococcus pyogenes, Enterococcus faecalis, Enter ococcus faecium, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Staphylococcus intermedius, Staphylococcus hyicus subsp. hyicus, Staphylococcus haemolyticus, Staphylococcus hominis, or Staphylococcus saccharolyticus.
[0144] [00144] In some embodiments, the infection that is treated or prevented comprises a bacterium that includes Pseudomonas aeruginosa, Pseudomonas fluorescens, Stenotrophomonas malto-philia, Escherichia coli, Citrobacter freundii, Salmonella typhimurium, Salmonella typhi, Salmonella paratyisidi, Salmonella paratyidi, Salmonella paratyidi, Salmonella , Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens, Acinetobacter calcoaceticus, Acinetobacter haemolyticus, Yersinia enterocolitica, Yersinia pestis, Yersinia pestis, Yersinia pestis, Yersinia pseudotubis parahaemolyticus, Helicobacter pylori, Campylobacter fetus, Campylobacter jejuni, Campylobacter coli, Vibrio cholerae, Vibrio parahaemolyticus, Legionella pneumophila, Listeria monocytogenes, Neisseria gonorrhoeae, Neisseria meningitidis, Moraxella, Bacteroides fragilis des vulgatus, Bacteroides ovalus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides eggerthii, or Bacteroides splanchnicus. EXAMPLES List of abbreviations
[0145] [00145] As used above, and throughout the description of the invention, the following abbreviations, unless otherwise indicated, will be understood to have the following meanings ACN acetonitrile BN benzyl BOC or Boc BOP benzotriazol-1-yl-oxitris (dimethylamino) phosphonium t-Bu tert-butyl Benzyl carbamate cbz Cyclohexyl Cy DBU 1,8-Diazabiciclo [5.4.0] undec-7-eno DCC dicyclohexylcarbodiimide DCM dichloromethane (CH2CI2) DIC 1,3-diisopropylcarbodiimide DEAD diethyl azodicarboxylate DIAD diisopropyl azodicarboxylate DIEA diisopropylethylamine DMAP 4- (N, N-dimethylamino) pyridine DMP reagent DMF dimethylformamide DMA N, N-Dimethylacetamide DME 1,2-Dimethoxy-ethane DMSO dimethylsulfoxide Dppf 1,1'-Bis (diphenylphosphino) ferrocene EDCI 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide HCl Equivalent Equivalent (s) Et ethyl Et2O diethyl ether EtOH ethanol EtOAc ethyl acetate HOAt 1-hydroxy-7-azabenzotriazole HOBT 1-hydroxybenztriazole HOSu N-hydroxysuccinamide HPLC high performance liquid chromatography LAH lithium aluminum anhydride Methylate me MeI methyl iodide MeOH methanol MOMCl methoxymethyl chloride MOM methoxymethyl MS mass spectroscopy NMP N-methyl-pyrrolidin-2-one Nuclear magnetic resonance NMR PyBOP Benzotriazol-1-yl-oxitris-pyrrolidine-phosphonium hexafluorophosphate SPHOS 2-Dicyclohexylphosphino-2 ', 6'-dimethoxybiphenyl TBD 1,5,7-triazabiciclo [4.4.0] -dec-5-ene RP-HPLC reverse phase high pressure liquid chromatography TBS ferc-butyldimethylsilyl TBSCl ferc-butyldimethylsilyl chloride TBTU O- (Benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium TEOC 2-T rimethylsilylethyl carbamate TFA trifluoroacetic acid Tf2O triflate anhydride TMG 1,1,3,3-Tetramethylguanidine THF tetrahydrofuran THP tetrahydropyran TLC thin layer chromatography XPHOS 2-Dicyclohexylphosphino-2 ', 4', 6'-triisopropylbiphenyl General Examples for the Preparation of Compounds of the Invention
[0146] [00146] The starting materials and intermediates for the compounds of this invention can be prepared by applying or adapting the methods described below, their obvious chemical equivalents or, for example, as described in the literature such as The Science of Synthesis, Volumes 1-8 . Editors E. M. Carreira et al. Thieme publishers (2001-2008). Reagent details and reaction options are also available through structure and reaction searches using commercial computer search engines such as Scifinder (www.cas.org) or Reaxys (www.reaxys.com).
[0147] [00147] Certain compounds of the invention (I) (SCHEME 1) are prepared from esters of boronic acid protected by corresponding functional groups (II) by treatment with a Lewis acid such as BCb, in a solvent such as dichloromethane, at a temperature between -78 ° C and 0 ° C, followed by an aqueous suppressor.
[0148] [00148] Alternatively, (I) is obtained from (II) by treating (II) with aqueous hydrochloric acid (around 3-5 Molar) in dioxane at a temperature between room temperature and 100 ° C.
[0149] [00149] The required boronic acid (II) esters are obtained (SCHEME 2) through the coupling of amine (III) with acid (carboxylic or sulfonic) (IV). This transformation is carried out first by activating the acid functionality as an acid chloride, anhydride or reactive ester (Va, Vb or Vc), followed by treating the activated substrate with (III) in a solvent such as DMF, DMA, NMP , THF or dichloromethane (or a mixture of these) at approximately room temperature, generally in the presence of a non-nucleophilic base such as 4-methyl-morpholine, triethylamine or diisopropylethylamine.
[0150] [00150] The formation of acid chloride (Va) involves the treatment of (IV) with a chlorinating agent such as thionyl chloride, phosphorus pentachloride or oxalyl chloride, in a solvent such as dichloromethane, in the presence of such a catalyst like DMF, around room temperature. In certain cases, DMF is also used as a cosolvent. The formation of anhydride (Vb) (Z is C = O) involves treatment of (IV) with a sterically hindered acid chloride or chloroformate, such as trimethylacetyl chloride or isopropylchloroformate, in an inert solvent such as dichloromethane, in the presence of a non-nucleophilic base, such as triethyl amine or diisopropylamine at room temperature or below. The formation of the activated ester (Vc) involves the treatment of (IV) with an activation reagent system such as EDCI, DCC / HOBt, HATU, BOP or TBTU reagents, in a solvent such as DMF, DMA, NMP or dichloromethane in room temperature or below (International Journal of Pharmaceutical Sciences Review and Research (2011), 8 (1), 108-119).
[0151] [00151] The requested acids (IV) are prepared by several different reaction sequences. Although there are common themes and strategies among the illustrative examples cited below, the selection of an appropriate reaction sequence (including protection group requirements) is dictated by the nature and disposition of the present functionality in the target molecule and, therefore, may involve obvious adaptations of the methods illustrated in order to be applied in a particular case.
[0152] [00152] In the case where Y1 is linked to CycA via an amine functionality, the required acids (IV) (SCHEME 3) are conveniently prepared from an appropriately substituted carbocyclic ketone (VI). For example, treatment of (VI) with a suitable amine (VII) in the presence of a reducing agent such as sodium tri-acetoxyborohydride, sodium cyanoborohydride or sodium borohydride in a solvent such as dichloromethane, 1,2-dichloro- ethane, THF, methanol, acetic acid or a mixture of these, at a temperature around room temperature provides ester (VIII). In the case where the use of a primary amine (VII, R5 = H) is called (VIII), it can also be prepared by treating an equimolar mixture of (VI) and (VII, R5 = H) with an acid of Lewis / desiccant, such as Ti (OEt) 4, in a solvent such as dichloromethane or 1,2-dichloroethane, at room temperature or above to provide the intermediate imine. This is followed by the reduction of the imine with sodium borohydride, in a solvent such as methanol, at a temperature between -78 ° C and room temperature.
[0153] [00153] The acid (IV) is obtained from the ester (VIII) by the formal hydrolysis of the ester functionality. The reaction conditions employed depend on the type of ester used. In the case of methyl, ethyl or other simple alkyl, hydrolysis is generally achieved by brief treatment with an aqueous base, such as sodium hydroxide or lithium hydroxide, in a solvent mixture of THF, water and methanol. However, other acid protecting groups can also be used, such as benzyl, allyl, 2-trimethylsilyl-ethyl or 2,2,2-trichloroethyl. In such cases, conversion of the ester to the corresponding acid is achieved using the standard deprotection products in the literature (Greene's Protective Groups in Organic Synthesis. Fourt Edition. John Wiley & Sons, Inc. 2006).
[0154] [00154] In certain cases, it is convenient to perform the reductive amination sequence using a keto-acid derivative (VI, R = H). In this case, the treatment of an equimolar mixture of keto-acid (VI, R = H) and amine (VII) with hydrogen gas in a solvent such as methanol, in the presence of a catalyst such as palladium on carbon, provides the acid (IV) directly.
[0155] [00155] In another approach to amino-linked systems (SCHEME 4), treatment of ketone (VI) with a reducing agent, such as sodium borohydride in methanol at around 0 ° C or L-selectride in THF, at a temperature between -78 ° C and room temperature it provides alcohol (IX). Treatment of alcohol (IX) with methanesulfonyl chloride or p-toluenesulfonyl chloride, in the presence of a non-nucleophilic base, such as triethylamine or DIEA, in a solvent such as dichloromethane or pyridine, at around 0 ° C, provides the corresponding sulfonate ester (X). The displacement of the sulfonate group with azide by treating (X) with sodium azide or a tetraalkylammonium azide in a solvent such as DMA, DMF, NMP, acetonitrile or DMSO, at a temperature between room temperature and 120 ° C, produces azide (XI). The reduction of azide with triphenylphosphine and water in THF around room temperature (Staudinger reaction) produces the primary amine (XII). Another derivatization of (XII), where appropriate, can be performed by reductive amination with an appropriate aldehyde or ketone, using the conditions already described to provide (XV).
[0156] Alternatively, the formation of the N-BOC derivative of (XII) through treatment with BOC2O, in the presence of a non-nucleophilic base such as triethylamine or DIEA, in a solvent such as dichloromethane, at room temperature, provides carbamate (XIII). Treatment of (XIII) with an alkyl halide or sulfonate in the presence of a base, such as sodium hydride, potassium carbonate or tetramethylguanidine, in a solvent such as DMF, DMA, NMP, THF, DMPU or ethanol (or a mixture of these), at room temperature or below, supplies (XIV). Cleavage of the BOC group with an acid, such as TFA in dichloromethane or HCl in dioxane, ethyl acetate or ether, around room temperature, provides the secondary amine (XV). Another derivatization of (XV), where appropriate, can be performed by reductive amination with an appropriate aldehyde or ketone, using the conditions already described to provide (VIII). The hydrolysis of (VIII) as already outlined produces (IV).
[0157] [00157] In the case where Y1 is a guanidine, the guanidine group is derived from the appropriate primary (XII) or secondary carbocyclic amine (XV) (SCHEME 5) through treatment with a reagent such as 1,3-Di-tert- butyloxycarbonyl-S-methylisothiourea, in a solvent such as DMF, (Synthesis, (2004), 37-42) or pyridine at room temperature or above, or through treatment with N, N! Bis- (BOC) -1H-pyrazol-1-carboxamidine in the presence of a base such as diisopropylethylamine, in a solvent such as DMF or DMA around room temperature. The selective cleavage of the ester functionality, as already described, provides the corresponding acid (IV).
[0158] [00158] Alternatively, the guanidinyl group can be introduced by treating an appropriate carbocyclic alcohol (IX) with a reagent such as Cbz-guanidine, in the presence of triphenylphosphine and diethyl-azo-dicarboxylati, in a solvent such as THF ( Mitsunobu conditions: Chemical Reviews, (2009), 109, 2551-2651) to supply (VIII) directly.
[0159] [00159] In the case where Y1 is an amidine linked to Cyc A through nitrogen, the required acids (SCHEME 6) are prepared from the appropriate primary (XII) or secondary (XV) amine by treatment with a suitable alkyl thioimidate, such as the 2-naptylmethylthioimidate derivative (XVI), in a solvent such as ethanol at a temperature between 0 ° C and room temperature (Tetrahedron Letters, (1997), 38 (2), 179-182) to provide (XVII ). The protection of amidine (XVII) as a carbamate derivative, such as BOC or Cbz under standard conditions, followed by beautiful selective ester hydrolysis provides (IV) acid.
[0160] [00160] In the case where Y1 is an amidine linked to CycA through carbon, the amidine functionality (SCHEME 7) is introduced by converting an appropriate carbocyclic ketone (XVIII) to the corresponding exocyclic nitrola (XIX) through treatment (XVIII) with toluenesulfonylmethylisocyanide (Journal of Organic Chemistry (1977), 42 (19), 3114-18) in the presence of a base such as COBut in a solvent such as DMSO or DME containing about 2% t-butanol or ethanol in a temperature between 0 ° C and 50 ° C. Treatment of (XIX) with HCl in methanol to form the corresponding imidate ester (XX) is followed by the reaction of this intermediate with an appropriate amine (R4R5NH), in a solvent such as methanol or THF around room temperature to provide the amidine (XXI). In certain cases, it is convenient to carry out the direct formation of amidine from nitrile (XIX) using a suitable methyl chloroaluminium amide, in a solvent such as toluene at around 80 ° C (Tetrahedron Letters, (1990), 31 ( 14), 1969-1972). In addition, in the case where R5 = H, amidine functionality can also be introduced by treating the appropriate carbocyclic nitrile (XIX) with hydroxylamine or an O-alkylhydroxylamine to provide N-hydroxyl- (or alkoxy) -amidine (XXII, R5 = OH, OR). This is followed by the removal of the protecting groups by catalytic hydrogenolysis to provide the amidine (XXIII). Selective acylation of amidine functionality in (XXIII) through treatment with BOC anhydride or Cbz chloride, as previously described, produces the primary alcohol (XXIV). The conversion of primary alcohol (XXIV) to the corresponding acid (IV) is performed using one of several oxidation protocols such as NaIO4 with RuCb catalytic in a mixture of water solvents / CCl4 / CH3CN in the ratio 3/2/2, at around room temperature (Journal of Organic Chemistry, (1981), 46 (19), 3936-8) or with pyridinium dichromate in DMF (Tetrahedron Letters, (1979). 20 (52), 399).
[0161] [00161] In certain cases, the primary alcohol (XXIV) is oxidized to the corresponding carboxylic acid using a two-step procedure, which involves initial oxidation with the aldehyde using a DMSO-based oxidizing system, such as Swern (Organic oxidation) Reactions. (1990), 39, 297-572.) Or through treatment with excess periodinane Dess Martin in a solvent such as dichloromethane around room temperature. The subsequent oxidation of the aldehyde is carried out by treatment with sodium chloride / sodium dihydrogen phosphate in the presence of tetramethylethylene, in a solvent such as t-butanol / water around room temperature (Journal of Organic Chemistry, (1980), 45, 4825).
[0162] [00162] In the case where Y1 is a methylene group substituted by nitrogen, the required acids (SCHEME 8) are prepared from the appropriate carbocyclic ketones (XVIII) by converting the ketone functionality, first, into the corresponding hydroxyl methyl derivative through treatment with an olefin reagent such as methyltriphenylphosphonium bromide in the presence of sodium hexamethyldisilazide, in a solvent such as THF at around 0 ° C. (Wittig reaction) or through treatment with lithium trimethylsilylmethane / CeCb at around 0 ° C to room temperature, in a solvent such as THF or ether (Peterson reaction) (Journal of Organic Chemistry, (1987) 52 ( 2), 281-3) or through the reaction with the Tebbe reagent modified by Petasis (dicyclopentadienyl-dimethyltitanium) in THF / toluene at around 60 ° C (Journal of the American Chemical Society, (1990), 112 (17) 6392 -6394 to supply (XXV) .This is followed by hydroboration oxidation of the exocyclic alkane in (XXV) with a reagent such as borane THF or an alkyl derivative, at around 0 ° C, in a solvent such as THF, followed by by oxidative preparation with hydrogen peroxide NaOH aq. to provide (XXVI) The conversion of hydroxymethyl (XXVI) to the functionalized amino-methyl derivative (XXVII) is performed by conversion to the corresponding tosylate, azide and primary amine as described above. Alternatively, the oxidation of (XXVI) to the aldehyde (XXVIII) followed by am reductive inhalation of (XXVIII) with an amine (R4R5NH), as already described, also provides amine (XXVII). The conversion of (XXVII) to the requested acid (IV) is performed by modifying the side chain as previously described.
[0163] [00163] In an alternative approach to systems where Yi is a nitrogen-substituted methylene group, CycA is a 5- to 7-membered carbocycle and Z is a carbonyl group, the required carboxylic acids (IV) are prepared from of an acrylamide derivative (XXIX) (SCHEME 9). (XXIX) is condensed, in a Diels Alder reaction, with an appropriately substituted diene, such as siloxy-diene (XXX), in an inert solvent such as toluene, xylene or DMA at a temperature between 70 o and 190 ° C to provide carbocyclic silylenol ether (XXXI).
[0164] [00164] The carbocyclic silylenol ether (XXXI) is then elaborated to supply several of the required carboxylic acids (IV) by applying the known transformations of the functional group. For example, (SCHEME 10), treatment of (XXXI) with N-phenyl-triflimide and CsF in a sealed system, using a solvent such as DME, around room temperature or below, provides the corresponding enol triflate (XXXII) ( Journal of the American Chemical Society, (2002) 124, 1129011291). The (XXXII) methoxy carbonylation with carbon monoxide / methanol in the presence of a non-nucleophilic base, such as triethylamine and a catalyst, such as Pd (OAc) 2, together with 1,3- (bis-diphenylphosphino) - Propane, in a solvent such as DMSO, at a temperature between 50 and 100 ° C, provides the unsaturated ester (XXXIII). The unsaturated ester (XXXIII) is reduced using a heterogeneous Pd, Rh or Pt catalyst, such as 10% Pd on carbon, under an atmosphere of hydrogen gas (1 to 4 atm), in a solvent such as ethyl acetate, methanol or THF (or a mixture thereof) at room temperature up to 70 ° C to provide the saturated ester (XXXIV). Alternatively, in certain cases, the unsaturated ester (XXXIII) can be reduced by treatment with excess magnesium, in a solvent such as methanol, around room temperature (Tetrahedron Letters, (1986); 27 (21), 2409- 2410) to provide (XXXIV).
[0165] [00165] In appropriate cases, the selective reduction of the amide functionality of the intermediate (XXXIV) to supply the corresponding amine (XXXV) (SCHEME 11) is carried out by treating (XXXIV) with a silane reducing agent, such as diphenylsilane and a rhodium catalyst such as rhodium-hydrocarbonyltriphenylphosphine, in a solvent such as THF, around room temperature (Tetrahedron Letters, (1998); 39 (9), 1017) or through treatment with 1,1,3, 3-tetramethyldisiloxane and a catalytic amount of chloroplatinic acid hydrate, in a solvent such as toluene, at a temperature between room temperature and 90 ° C (Journal of the American Chemical Society, (2009); 131 (41), 15032- 15040).
[0166] [00166] The ester functionality in (XXXV) is hydrolyzed to provide the required acid (IV) for brief treatment with a base such as sodium hydroxide or lithium hydroxide, in a solvent such as THF / methanol / water, around room temperature.
[0167] [00167] Alternatively, the reduction of amide functionality in (XXXIV), with the concomitant reduction of the ester group, to provide (XXXVI), is carried out by treating (XXXIV) with a hydride reducing agent, such as lithium aluminum hydride, in an ethereal solvent such as THF, diethyl ether, dimethoxy-ethane or methyl-butyl-ether, at a temperature between 0 ° C and 65 ° C. The conversion of the resulting amino alcohol (XXXVI; R4, R5 = H) to the corresponding acid (IV), is carried out through the derivatization of amine and the oxidation of the primary alcohol as already described.
[0168] [00168] Carboxylic acid homologated by a corresponding carbon (IV) where Cyc A is a six-membered ring, L, M = bond, m = 0, n = 1, Y1 = (R4R5NCH2) (SCHEME 12), is also prepared from carbocyclic silyl ether (XXXI). Treatment of (XXXI) with potassium carbonate in methanol around room temperature or with tetra-n-butylammonium fluoride hydrate in a solvent such as THF (and where appropriate, capped with an equimolar amount of acetic acid) provides ketone ( XXXVII). Treatment of (XXXVII) with a trialkyl-phosphonoa-ketate, such as triethylphosphonoacetate, in a solvent such as THF, in the presence of a base such as sodium hydride, at a temperature between about -5 ° C and room temperature, provides the corresponding α, β-unsaturated ester (XXXVIII) (Liebigs Annalen / Recueil, (1997), 7, 1283-1301.). Reduction of alkane and amide functionality in (XXXVIII) and subsequent processing (ester hydrolysis or alcohol oxidation), analogous to that described above provides the required acid (IV).
[0169] [00169] Carboxylic acid (IV) where Cyc A is a six-membered ring, L, M = bond, m = 0, n = 2, Y1 = (R4R5NCH2) (SCHEME 13) is prepared from alcohol (XXXVI ) through oxidation with the corresponding aldehyde (XXXIX) under standard conditions followed by a Horner-Wadswort-Emmons reaction, as described above, to provide the α, β-unsaturated ester (XL). The reduction of the alkane in (XL) and the ester hydrolysis analogous to that described above provides the required acid (IV).
[0170] [00170] The carboxylic acid (IV) where Cyc A is a six-membered ring, L = O, M = bond, m = 0, n = 1, Y1 = R4R5NCH2) (SCHEME 14) is prepared from ketone (XXXVII) through treatment with a reagent such as sodium borohydride in methanol at around 0 ° C or L-selectride in THF, at a temperature between -78 ° C and room temperature to provide the alcohol (XLI). Condensation of (XLI) with ethyl diazoacetate in the presence of a catalyst such as Rh (acac) dimer in a solvent such as dichloromethane, provides the alkoxyacetate derivative (XLII). Selective amide reduction and ester hydrolysis of (XLII) as already described produces (IV).
[0171] [00171] Carboxylic acid (IV) where Cyc A is a six-membered ring, L = NR4, M = bond, m = 0, Y1 = (R4R5NCH2) (SCHEME 15) is prepared from ketone (XXXVII) through treatment with an appropriate amino ester (XLIII) under the reductive amination conditions already described to produce (XLIV). Selective amide reduction and ester hydrolysis of (XLIV), as already described, provides the required acid (IV).
[0172] [00172] The seven-membered carbocyclic systems (IV) (SCHEME 16) are accessed by expanding the carbocyclic silylenol ether ring (XXXI).
[0173] [00173] For example, the treatment of (XXXI) with a synthetic carbene equivalent, such as Simmons Smith's reagent, followed by the gentle oxidation of the resulting cyclopropane, using a reagent such as FeCb (Journal of Organic Chemistry, (1985) , 50 (4), 531-534) or CAN / NaOAc (Organic Letters, (2007), 9 (7), 1323-1326), provides the seven-member enone (XLV). Treatment of (XLV) with a silane, such as PhMe2SiH, in the presence of a catalyst, such as the Wilquinson (Ph3P) 3RhCl catalyst, pure or in a solvent such as benzene or toluene, provides the expanded ring silyl ether (XLVI). This seven-membered carbocycle is processed in a manner directly analogous to this six-membered ring congener (XXXI) described above, to supply the required acids (IV, CycA = heptane).
[0174] [00174] In certain cases, it is convenient to prepare carbocyclic systems (IV, CycA = cyclopentane) (SCHEME 17) from ketone (XXXI) by Favorskii rearrangement of the corresponding α-halo-ketone (XLVII) (Current Organic Chemistry ( 2005), 9 (17), 1713-1735). For example, treatment of (XXXI) with a halogenating reagent, such as bromine, NCS, or pyridinium tribromide in an inert solvent such as dichloromethane or hexane, at a temperature between -78 ° C and room temperature, provides the α-halo-ketone (XLVII). The application of Favorskii's rearrangement conditions in (XLVII) (slow addition to a suspension of sodium methoxide in ether, around room temperature) then provides the contracted ester-substituted carbocycle (XLVIII).
[0175] [00175] This intermediate is processed in a manner directly analogous to its six-membered ring counterpart (XXXIV) described above, to supply the required acids (IV).
[0176] [00176] In certain cases carbocyclic systems (IV, CycA = cyclopentane) are prepared through a cycloaddition reaction between an acrylamide derivative (XXIX) and 2 - ((trimethylsilyl) methyl) allyl acetate (XLIX), in a solvent such as THF, toluene or xylene, in the presence of a catalyst such as bis (diphenylphosphinopropane) / palladium acetate, at a temperature between 70 ° and 160 ° C (Journal of the American Chemical Society. (1979), 101 (21) , 6429-6431) to provide (L). The exocyclic olefin in (L) is oxidatively cleaved by treatment with catalytic amounts of osmium tetroxide (Org. Synth. Oxid. Met. Compd. (1986), 633-93. Publisher: Plenum, New York) in the presence of a co-oxidant such as N-methyl morpholine N-oxide, in a solvent such as tert-butanol / water to produce the corresponding dihydroxy derivative (LI). This diol is then oxidatively cleaved using sodium periodate, in a solvent such as THF / water, around room temperature, to provide (LII).
[0177] [00177] The carbocyclic ketone (LII) is then processed in a manner directly analogous to this six-membered ring congener (XXXVII) to provide the corresponding acid (IV).
[0178] [00178] In the case where Y1 and Y2 are both methylene groups substituted by nitrogen in such a way that Y1 and Y2 are positioned close to the carbocycle, the required acids (IV) are prepared as illustrated above (SCHEMES 9 to 18) except starting with a fumaric or maleic acid diamide instead of an acrylamide.
[0179] [00179] In the case where CycA is a 5, 6 or 7 membered carbocycle in which Y3 and Y4 taken together form a ring, the required carboxylic acids (IV) (SCHEME 19) are prepared by the methods described above, except for a siloxide. cyclic diene such as (LIII) is used in the Diels Alder reaction to provide bicyclic silylenol ether (LIV). The starting diene structures are prepared from the corresponding α, β-unsaturated ketone (LV) by treating the appropriate enone with trialkylsilyltriflate in the presence of a base such as triethylamine, proton sponge or DBU in a solvent such as ether a 0 ° C or above.
[0180] [00180] In the case where CycA is a 5, 6 or 7 membered carbocycle, M = bond, O, NR4, Y1 and Y2 are each linked to CycA via a nitrogen atom and Y1 and Y2 are positioned close together in the carbocycle, the required acids (IV) (SCHEME 20) are prepared from the appropriate cyclic olefins (LVI). For example, treatment of (LVI) with sodium azide, in the presence of a mild oxidant, such as Mn (OAc) 3 (H2O) 2 and an acid such as acetic acid or trifluoroacetic acid, in a solvent such as acetonitrile, in a temperature between -30 ° C and 0 ° C, provides diazide (LVII) predominantly in the transisomer configuration (Synthetic Communications, 28 (10), 1913-1922; 1998). The subsequent reduction of bis-azide by treatment with a reducing agent, such as triphenylphosphine, in a solvent such as THF, followed by the in-situ hydrolysis of the intermediate aza-phosphorane by adding excess water produces the bis-amine ( LVIII). In certain cases, the reduction of bis-azide is also achieved through treatment with hydrogen gas in the presence of a catalyst such as palladium on carbon in a solvent such as THF or methanol.
[0181] [00181] This primary bis-amine (LVIII) is protected as a BOC, Cbz or other suitable N-protected derivative (Greene's Protective Groups in Organic Synthesis; 4t Edition: John Wiley & Sons, Inc., 2006). For example, treatment of (LVIII) with an appropriate anhydride or chloroform, in the presence of a base such as triethylamine, in a solvent such as THF or dichloromethane, around room temperature, provides the carbamate intermediate (LIX). Where appropriate, carbamate is further derivatized by treatment with a suitable alkylating agent, in the presence of a base, such as K2CO3, in a solvent such as DMF, DMA, or acetonitrile, to provide (LX). The removal of the carbamate protecting group, followed by treatment of the resulting secondary amine with an aldehyde or ketone in the presence of a reducing agent such as sodium borohydride, sodium cyanoborohydride or sodium triacetoxyborohydride, in a solvent such as dichloromethane, 1 , 2-dichloroethane, methanol or THF, around room temperature, provides (LXI). Hydrolysis of the (LXI) ester, as described above, produces the desired acid (IV).
[0182] [00182] Alternatively, treatment of the appropriate cyclic olefin (LVI) (SCHEME 21) with an oxidant such as meta-chloro-perbenzoic acid in a solvent such as dichloromethane at around 0 ° C, provides the corresponding cyclic epoxide (LXII) . The opening of the epoxide ring through treatment with sodium azide and ammonium chloride in a solvent such as ethanol, polyethylene glycol, or DMF / water at a temperature between room temperature and 80 ° C, provides the trans hydroxyl azide (LXIII). The reaction of (LIII) with methanesulfonyl chloride in pyridine at around 0 ° C produces the mesylate (LXIV). Treatment of (LXIV) with tetrabutylammonium azide in a solvent such as toluene, provides cis-oriented bis-azide (LXV). The processing of the intermediate (LXV) is carried out as previously described to provide acids (IV).
[0183] [00183] Where appropriate, cyclic olefins (LVI) (SCHEME 22) are prepared from acyclic starting materials through olefin metathesis. For example, treatment of ester (LXVI) with a strong base, such as LDA in a solvent such as THF, THF / DMPU or DME at a temperature between -78 ° C and 0 ° C, forms the corresponding lithium enolate. This lithium enolate is treated with a halo-alkyl olefin suitable to produce bis-olefin (LXVII). The treatment of (LXVII) with that of a range of Grubb's or Schrock metathesis catalysts (Tetrahedron, (2012), 68 (2) 397-421: Organic Letters, (2007), 9 (23), 4885-4888: Tetrahedron, (2004), 60, 7117-7139) in a solvent such as dichloromethane, at room temperature or above or, in an aqueous solution of dimethyl ether PEG500, provides the cyclic olefin (LVI, M = bond, n = 0) .
[0184] [00184] It should be noted that the cyclic silyl enol ethers or alkyl enol ethers corresponding to (LVI) which are precursors of cyclic ketones, are also conveniently prepared by this approach using a suitable silyl ether or acyclic alkyl ether in the closed ring metathesis reaction (Tetrahedron Letters, (2001), 42 (45), 8023).
[0185] [00185] The subsequent transformations of the standard functional group into the exocyclic alkoxycarbonyl group of (LVI, M = bond, n = 0) provide a range of homologous cyclic olefins substituted by alkoxycarbonyl-alkyl (SCHEME 23). For example, the reduction of ester functionality through treatment with a reducing agent such as DIBALH, in a solvent such as toluene, THF or dichloromethane, at a temperature between -78 o and 0 ° C, provides the primary alcohol (LXVIII) . The oxidation of (LXVIII) with that of a range of oxidants described above, (such as periodinane Dess-Martin), in a solvent such as dichloromethane, produces the corresponding aldehyde (LXIX). Wittig's olefin (LXIX) using a triphenylphosphoranilidine-acetate ester (LXX), in a solvent such as toluene or THF, at a temperature between room temperature and 80 ° C, provides the unsaturated ester (LXXI). The selective reduction of the conjugated double bond in (LXXI) in the corresponding saturated ester (LVI, n = 2) is carried out by treatment with a tape of magnesium, in a solvent such as methanol, around room temperature.
[0186] [00186] Alternatively, the treatment of alcohol (LXVIII) with p-toluenesulfonyl chloride, in a solvent such as pyridine or dichloromethane, in the presence of a base such as triethylamine or DMAP, at a temperature between -20 ° C and the temperature environment, provides the tosylate derivative (LXXII). The displacement of the tosyl group in (LXXII) through treatment with sodium cyanide in DMF, DMA or DMSO, at a temperature between room temperature and 160 ° C, provides nitrile (LXXIII). The solvolysis of this nitrile through treatment with HCl in an appropriate alcohol (such as methanol or ethanol) produces (LVI, M = bond, n = 1).
[0187] [00187] The closed ring metathesis can be used to provide access to (LVI, M = O or NR4) through the use of bis alkane substrate substituted by alkoxy or amino (SCHEME 24). For example, coupling an appropriately substituted aldehyde or imine (LXXIV) with a suitable Grignard provides the secondary alcohol (sulfinamine) (LXXV). The alkylation or reductive amination of (LXXV) under standard conditions provides (LXXVI). Exposure of (LXXVI) to closed ring metathesis conditions, as described above, provides carbocyclic alkane (LVI, M = O, NR4).
[0188] [00188] In the case where CycA is a 4- to 7-membered carbocycle, Y1 is a group of 1-amino-t-amino-alkyl (t = 2, 3, 4), the required acids (IV) (SCHEME 25) they are generally prepared by sequential installation of the amino functionality on the hydroxyl-substituted or substituted carbonoxy carbon scaffold. For example, the selective reduction of ester functionality in (LXXVIII) in the corresponding alcohol (LXXIX) is carried out by treatment with a reducing agent, such as borane: THF, in a solvent such as THF, around room temperature. The oxidation of primary alcohol to the corresponding aldehyde (LXXX) is achieved by that of a wide range of standard oxidation protocols such as Swern oxidation or Dess-Martin periodane oxidation. The aldehyde is then treated with an olefin-substituted Grignard alkyl reagent, in a solvent such as THF, diethyl ether or DME at a temperature between about -50 ° C and room temperature, to provide the secondary alcohol (LXXXI). In certain cases, where the required olefin substituted alkyl is an allyl (t = 1), condensation with the aldehyde (LXXX) can also be carried out using an allyl boronate or allyl silane in the presence of a Lewis acid such as TiCU or BF3 etherate in a solvent such as dichloromethane at a temperature between -78 ° C and room temperature. The olefin-substituted secondary alcohol (LXXXI) is then converted to the corresponding amine by treatment with p-toluenesulfonyl chloride in the presence of a base such as triethylamine or DMAP, in a solvent such as dichloromethane or pyridine, at a temperature between -20 ° C and room temperature to provide the corresponding tosylate (LXXXII). The tosylate is then treated with an azide salt such as sodium azide, tetrabutylammonium azide in a solvent such as DMF, DMA or DMSO at a temperature between room temperature and 160 ° C to produce the corresponding azide (LXXXIII). This intermediate is reduced to the primary amine (LXXXIV) and subsequently derivatized as described above to provide (LXXXV). Alternatively, the introduction of the primary amino functionality is carried out by converting the secondary alcohol (LXXXI) to the corresponding ptalimide (LXXXVI) under the Mitsunobu conditions (Chemical Reviews, (2009) 2551-2651) followed by the deprotection of the ptalimide through treatment with excess hydrazine in a solvent such as ethanol at room temperature or above, to provide the primary amine (LXXXIV).
[0189] [00189] The installation of a second amino functionality in (LXXXV), is performed through the oxidative cleavage of the olefin as previously described, to supply the aldehyde (LXXXVII). This intermediate is converted to the requested amine (LXXXVIII) through direct reductive alkylation or by reduction in the primary alcohol using a reducing agent such as sodium borohydride in a solvent such as THF / methanol at a temperature between -78 ° C and 0 ° C, followed by derivatization of the resulting primary alcohol (by means of the corresponding tosylate, azide and primary amine) as described above. The processing of the side chain of (LXXXVIII) as previously described provides (IV).
[0190] [00190] In the particular case where Y1 is an optionally substituted 1-amino-2-amino-ethyl group, the installation of the diamine functionality can be performed by direct amination of the appropriate exocyclic alkane (Mn (OAc) 3 / NaN3 / TFA then reduction). The requested exocyclic olefin is prepared by the Wittig, Peterson or Tebbe type olefin (LXXX) as already described.
[0191] [00191] In the case where CycA is an optionally substituted 4- to 7-membered carbocycle, Y1 is an optionally substituted t-amino-alkyl group (t = 1, 2, 3, 4), Y2 is an optionally substituted amine, and Y1 and Y2 are both bonded on the same carbon of the CycA ring, the required acids (IV) (SCHEME 26) are prepared from the appropriate carbocyclic ketone precursor (LXXXIX) through treatment with t-butylsulfinamine (Chemical Reviews, (2010 ), 110 (6), 3600-3740), typically in a solvent such as THF or methanol, in the presence of Ti (OEt) 4, at a temperature between room temperature and 60 ° C. The resulting t-butylsulfinimine is then condensed with an appropriate organometallic, such as an olefin-substituted alkyl Grignard (or where appropriate, a CeCl3 modified Grignard reagent) in a solvent such as THF, ether, dichloromethane or toluene at a temperature between -60 o and 0 ° C to provide the sulfinamine-substituted carbocycle (XC). The removal of the sulfinyl group is carried out by treatment with an acid such as trifluoroacetic acid in a solvent such as dichloromethane at around room temperature to produce the corresponding primary amine (XCI, R4, R5 = H). The primary amine is derivatized or protected, as appropriate, by the methods described above. The processing of the alkane functionality to provide the appropriate amine is carried out by oxidative cleavage (OsO4 / NMO then NaIO4), then the reductive amination of the resulting aldehyde to provide (XCII). Alternatively, aldehyde hydride reduction and alcohol derivatization, as described above, also provides (XCII). The conversion of (XCII) to the requested acid is performed by processing the precursor group of the side chain of acid Y as already described.
[0192] [00192] In the specific case where t = 1, (IV) is prepared from ketone (LXXXIX) through a Strecker reaction (Synthesis, (2007), 12301234. Organic Letters, (2008), 10, 1509-1512) followed by reduction of the resulting nitrile group. The subsequent processing of this intermediate to supply (IV) is carried out as already described.
[0193] [00193] In a variation of the above system where CycA is an optionally substituted 4- to 7-membered carbocycle, Y1 is a t-amino-alkyl group (t = 2, 3, 4), Y2 is an amine, Y1 and Y2 are both attached to the same carbon of the CycA ring and Y1 and Y2 are connected to each other through their substituents (by a group of CH2), the required acids (IV) (SCHEME 27) are prepared from the intermediate (XCI) through treatment with OsO4 in a solvent such as acetone / water in the presence of an oxidant such as N-methyl-morpholine N-oxide to produce the corresponding diol. This intermediate is treated with sodium periodate in THF / water to form the truncated aldehyde (XCIII). (XCIII) is then condensed with an α-phosphonoglycine ester derivative, such as the BOC-protected trimethyl α-phosphono-glycine ester, in the presence of a base such as potassium t-butoxide in a solvent such as dichloromethane in a temperature between -78 ° C and -50 ° C to provide the unsaturated ester (XCIV) (Tetrahedron, (2001), 57, 6463). (XCIV) is selectively hydrogenated using a rhodium phosphine-cationic phosphite catalyst to provide the corresponding saturated BOC-TCEOC protected amino ester. This intermediate is deprotected under standard conditions (Zinc / acetic acid in THF) to produce the BOC-protected amino ester (XCV).
[0194] [00194] The cyclization of the amino ester, in the presence of a base such as DBU, in a solvent such as toluene, at a temperature between room temperature and 110 ° C, provides the lactam (XCVI). Selective reduction of tertiary amide in (XCVI) using a silane reducing agent in the presence of rhodiohydrocarbonyltriphenylphosphine, or chloroplatinic acid, as described above, followed by amine derivatization, provides (XCVII). Subsequent processing of the side chain Y, as previously described, supplies the required acids (IV).
[0195] [00195] In the case where CycA is an optionally substituted 4- to 7-membered carbocycle, Y1 is an optionally substituted t-amino-alkyl group (t = 2, 3, 4), Y2 is an optionally substituted amine, and carbon of the CycA ring linked to Y1 and the carbon of the CycA ring linked to Y2 are positioned close together, the required acids (IV) (SCHEME 28) can be prepared from the appropriate cyclic ketone (XCVIII). For example, in the case where t = 2, treatment of (XCVIII) with a trialkylsilyl triflate in the presence of a base such as triethylamine in a solvent such as ether, provides the corresponding silyl enol ether (XCIX). The reaction of this intermediate with a nitro-olefin (R6R7C = CR6NO2) in the presence of a Lewis acid such as TiCl4 / (TiOiPr) 4 or SnCÍ4 in a solvent such as dichloromethane at a temperature between -78 ° C and room temperature, supplies the cyclic ketone substituted by nitro-alkyl (C) (Journal of the American Chemical Society, 106 (7), 2149-56; 1984; Helvetica Chimica Acta, 82 (11), 18291842; 1999; Canadian Journal of Chemistry, 65 (4), 836-50; 1987). The hydride reduction of this intermediate using a reagent such as lithium aluminum hydride in a solvent such as THF at a temperature between -20 ° C and reflux, provides the amino alcohol (CI). The amino group of (CI) is selectively derivatized as described above to provide (CII). The installation of the second amino group is carried out by derivatizing carbocyclic alcohol (CII) using the methods described above to supply (CIII). The removal of the benzyl ether protecting group and the oxidation of the resulting primary alcohol are carried out as described previously to produce the required carboxylic acid (IV).
[0196] [00196] For the case where t = 1 (SCHEME 29), the installation of the appropriate side chain functionality is achieved by the reaction of silylenol ether (XCIX) with an imine salt (where appropriate generated in situ from N , O-acetal and trimethylsilyltriflate) in a solvent such as dichloromethane at a temperature between -78 ° C and room temperature to provide the amino ketone (CIV). The processing of the ketone in (CIV), as previously described, provides the diamino functionalized scaffold (CV). Therefore, the removal of benzyl ether and the oxidation of the resulting primary alcohol in the usual manner produce the desired acid (IV).
[0197] [00197] For the case where t = 3 (SCHEME 30), the installation of the appropriate side chain functionality is achieved by the reaction of silylenol ether (XCIX) with methyl lithium, in a solvent such as THF, at a temperature between -20 ° C and 0 ° C, to generate the corresponding lithium enolate regioselectively. This is followed by treatment of the enolate with an electrophile such as an acrylamide (or acrylonitrile) to provide (CIV). Processing the ketone as already described provides the amine-functionalized amide (CVII). Reducing the amide with a reducing agent such as lithium aluminum hydride in a solvent such as THF at a temperature between -10 ° C and reflux, produces the diamine (CVIII). This intermediate is then processed, as described above (benzyl removal treatment, then oxidation), to provide the required acid (IV).
[0198] [00198] In the case where A is an optionally substituted 4- to 7-membered carbocycle, Y1 is an optionally substituted r-amino-alkyl group (t = 2, 3, 4), Y2 is an optionally substituted amino-methylene, and both Y1 and Y2 are bonded to the same carbon as the ring of A, the required acids (IV) (SCHEME 31) are prepared from carbocyclic ketone (CIX). For example, the conversion of (CIX) to the corresponding exocyclic nitrile (CX) is performed by treatment with TOSMIC and base as previously described. Treatment of (CX) with a base such as LDA, LHMDS or NaHMDS in a solvent such as THF and an imine, such as i-butylsulfinimine (Chemical Reviews, (2010), 110 (6), 3600-3740) provides the nitrile substituted by i-butylsulfinamino-methyl (CXI). The removal of the sulfinyl group by treatment with an acid such as trifluoroacetic acid in dichloromethane followed by derivatization of the primary amine as described above, provides the amino nitrile (CXII). The reduction of nitrile (CXII) with lithium aluminum hydride, as described above, produces the primary amine (CXIII). This amine is further derivatized, where appropriate, as described above (this may also include temporary derivatization using an easily removable protecting group) to provide (CXIV). Subsequent processing of the exocyclic benzyl ether in (CXIV), using the usual procedures, supplies the required acid (IV).
[0199] [00199] In the case where CycA is a 6 or 7 membered carbocycle, Yi is an optionally substituted r-amino-alkyl group (t = 1, 2, 3), Y2 is an optionally substituted amine and the ring carbon in CycA linked to Y1 and the carbon of the ring in CycA linked to Y2 are separated by a methylene, the required acids (IV) (SCHEME 32) are prepared from an appropriately substituted carbocyclic enone such as (CXV). For example, in the case where t = 1, an appropriately functionalized carbon unit is installed by treating the carbocyclic enone (CXV) with a cyanate reagent system such as trimethylsilyl cyanide or hydrogen cyanide in the presence of such a base such as KF, Cs2CO3, Bu4NF or tetramethylguanidine in a suitable solvent, such as methanol, THF, DMA or acetonitrile to provide (CXVI). The introduction of nitrile can also be carried out by treating the enone (CXV) with diethyl aluminum cyanide in a solvent such as benzene, toluene or dichloromethane, at a temperature between about -20 ° C and room temperature. Subsequent derivatization of carbocyclic ketone (CXVI), (by means of reductive amination, conversion to azide; reduction of azide and other derivatization of the resulting primary amine), as previously described, provides the carbocycle substituted by 1-amino-3-cyano (CXVII ). This intermediate is converted to the aminomethyl derivative required by the reduction of nitrile and functionalization of the resulting primary amine (CXVIII) as already described to supply (CXIX). Side chain processing in the usual manner provides (IV).
[0200] [00200] In the case where t = 2 or 3 (SCHEME 33), the appropriately functionalized carbon scaffold is prepared by treating the carbocyclic enone (CXV) with an allyl silane in a non-coordinating solvent such as dichloromethane in the presence of a Lewis acid such as TiCl4, SnCl4, or BF3 etherate, at a temperature between -78 ° C and room temperature to produce the olefin-functionalized carbocyclic ketone (CXX). The processing of the ketone functionality in (CXX), as already described, produces (CXXI). The oxidative cleavage of the olefin in (CXXI) and processing the resulting aldehyde into the corresponding amine, as already outlined, provides (CXXII). This intermediate is then converted to the required acid (IV, t = 2) by processing the side chain in the usual manner. Alternatively, hydroboration of the olefin (CXXI) through treatment with a borane, such as 9-BBN, in a solvent such as THF at around 0 ° C, followed by the oxidative preparation (NaOH / H2O2) provides (CXXIII). Subsequent processing of (CXXIII) as already described, establishes the amine functionality to provide (CXXIV). The side-chain processing of this intermediate produces acid (IV, t = 3).
[0201] [00201] In the case where A is an optionally substituted 6 or 7 membered carbocycle, Y1 is an optionally substituted amine, Y2 is an optionally substituted amine and the ring carbon in CycA is attached to Y1 and the ring carbon in CycA is attached to Y2 are separated by a methylene, the required acids (IV) (SCHEME 34) are prepared from an appropriately substituted carbocyclic enone such as (CII) by treatment with a suitable amine in the presence of a catalyst such as RuCb, Cu ( acac) 2, FeCl3, Ce (NH4) NO3 Pd (acac) 2NH4PF6 in a solvent such as water or polyethylene glycol (Green Chemistry, (2006), 8 (4), 356-358; Synthesis, (2005), (13 ), 2129-2136; Helvetica Chimica Acta, (2004), 87 (6), 1522-1526; Advanced Synthesis & Catalysis, (2005), 347 (6), 763-766; Synthetic letters, (2006), (10 ), 1549-1553) at a temperature between room temperature and 80 ° C to provide β-amino ketone (CXXV). Subsequent processing of the ketone functionality as described above produces the diamine (CXXVI). The conversion of (CXXVI) to acid (IV) is achieved by side chain processing as previously described.
[0202] [00202] In the case where Y1 is an optionally substituted amino-alkyloxy, the required acids (IV) (SCHEME 35) are prepared by treating an appropriate carbocyclic alcohol (CXXVII) with an alkyl halide substituted by ptalimido or azido in the presence of a base, such as sodium hydride, in a solvent such as DMF, DMA, DMSO, at a temperature between 5 ° C and 80 ° C (a catalyst such as tetrabutylammonium iodide can also be used) to provide the alkoxy-carbocycles replaced by corresponding latent amino (CXXVIII and CXXIX) respectively. The unmasking of the latent amine by treating (CXXVIII) with hydrazine in ethanol, or treating (CXXIX) with triphenylphosphine and water in THF, provides the corresponding primary amine (CXXX). The derivatization of amine as previously described produces (CXXXI). The processing of (CXXXI) by the standard method provides the required acid (IV).
[0203] [00203] Alternatively (SCHEME 36), the treatment of alcohol (CXVII) with a reagent system such as iodine / imidazole / Ph3P or NBS / Ph3P or with an appropriate sulfonyl chloride (or anhydride) in the presence of a base such as pyridine or triethylamine in a solvent such as THF or dichloromethane, provides the corresponding iodide, bromide or sulfonate (CXXXII). Reaction of (CXXXII) with an appropriate alcohol in the presence of a base such as sodium hydride, in a solvent such as THF, DMF, DMA or TBTU, or a mixture of these, provides the carbocycles substituted by alkoxy (CXXVIII) and ( CXXIX). The conversion of these intermediates to (IV) is carried out as already described.
[0204] [00204] Formula (I) provides the case where Y1 and Y2, taken together with the carbon atom or carbon atoms to which they are attached, form an optionally substituted carbocycle or optionally substituted heterocycle. A specific example of this provision (SCHEME 37) is where CycA is a cyclohexane, Y1 is a group of (N-methyl-guanidinyl) -methyl. Y2 is a methyl group. Y1 and Y2 are positioned close together on CycA and Y1 and Y2 are linked together by the formal fusion of two methyl groups to form a substituted piperidine. In this particular case, the requested acids (IV) are prepared from an appropriate piperidinone such as (CXXXIII). For example, treatment of (CXXXNI) with α-methyl benzylamine (Bioorganic and medicinal Chemistry Letters, (2008), 18 (4), 1312-1317) and an appropriate methyl-vinyl ketone derivative in toluene followed by cyclization with methoxide sodium in methanol provides the bicyclic ketone (CXXXIV). Hydrogenation of (CXXXIV) using a catalyst such as palladium on carbon in a solvent such as methanol and the installation of the guanidinyl group as already described, produces the ketone (CXXXV). The processing of the ketone (CXXXV) as already outlined provides the required acids (IV, M = binding; n = 1.2). Alternatively, the reduction of (CXXXV) with a hydride reducing agent such as sodium borohydride in methanol at a temperature between -78 ° C and 0 ° C, provides the corresponding alcohol which is treated with ethyl diazoacetate and a catalytic amount of Rh dimer (acac) 2 in a solvent such as dichloromethane to provide the ester (CXXXVI). Saponification of (CXXXVI) by brief treatment with lithium hydroxide in THF / methanol / water provides the acid (IV, M = O, n = 1).
[0205] [00205] In the case where Z is a sulfonyl group (SCHEME 38), the requested sulfonic acid is prepared from the corresponding activated carboxylic acid (V) by treatment with sodium hydroxythiopyridone in a solvent such as dichloromethane, around room temperature , to produce the Barton ester intermediate (CXXXVII). (CXXXVII) is treated with iodoform in CCI4 under a tungsten UV lamp around the reflux temperature to provide the decarboxylative iodination product (CXXXVIII) (Journal of Organic Chemistry, 75 (19), 6489-6501; 2010) . Alternatively, the treatment of acid (IV) with iodine-benzene diacetate and iodine in CCl4, under a tungsten UV lamp, around the reflux temperature (Journal of Organic Chemistry, (1986), 51, 402), provides (CXXXVIII) directly. The treatment of (CXXXVIII) with sodium sulfite in ethanol, isopropanol or aqueous acetone, at a temperature between 60 and 90 ° C, followed by acidification, produces sulfonic acid (IV). Alternatively, the treatment of (CXXXVIII) with thiourea in acetone, around 60 ° C, provides the derivative of isothiouronium salt (CXXXIX) (Synthetic Letters, (2010), 7, 1037). The cleavage of (CXXXVIII) with aq. supplies thiol (CXL). Treatment of (CXL) with perfromic acid (formic acid and aqueous H2O2 at around 0 ° C to room temperature) provides (IV).
[0206] [00206] The following preparations of Formula I or Formula Ia compounds and intermediates are provided to allow those of skill in the art to clearly understand and practice the present invention. They should not be considered as limiting the scope of the invention, but only as illustrative and representative of it. EXAMPLE 1: (R) -3- (trans-4- (aminomethyl) cyclohexanecarboxa-mido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin- 8-carboxylic
[0207] [00207] To anhydrous CH2Cl2 (0.61 mL, 9.4 mmol) in THF (20 mL) under Argon at -100 ° C (MeOH / N2 Liq.) Was added n-BuLi (2.7 mL, 2, 5 M in hexane) by dropping and the reaction mixture was stirred at the same temperature for 30 min. A solution of THF (5 mL) of 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,02, 6] dec-4-ylmethyl) -benzoic (2.37g, 5.92 mmol) was added over a period of 10 min. After 20 min, the cooling bath was removed and the reaction mixture was slowly warmed to 0 ° C and stirred at the same temperature for 1 h. The reaction mixture was then cooled to -78 ° C, LHMDS (8.0 mL, 1M in THF) was added slowly and the resulting reaction mixture was stirred while warming to room temperature gradually overnight. Anhydrous MeOH (0.29 mL, 7.1 mmol) was added at -10 ° C, the reaction was stirred at the same temperature for 1 h and then at room temperature for 1 h.
[0208] [00208] In a separate flask containing 0.386 g of 4 - ((tert-butoxycarbonylamino) methyl) cyclohexanecarboxylic acid (1.5 mmol), anhydrous CH2Cl2 (12 mL) was added. To this reaction mixture was added NMM (0.22 ml, 2 mmol), followed by HATU (0.570 g, 1.5 mmol). DMF (1 mL) was added and the resulting solution was stirred at room temperature (RT) for 1 h, during which time a portion of the above reaction solution (1.5 mmol) was added to the flask and the reaction was stirred for 2 hours. H. The reaction was quenched by the addition of water (30 ml) and the aqueous phase extracted with EtOAc (3X50 ml). The organic phase was washed with brine, dried over Na2SO4, and concentrated in vacuo to provide the crude product, which was purified by silica gel scintillation chromatography (hexane / EtOAc, 2: 1 to 1: 2) to provide the product ( 200 mg, 20%). ESI-MS m / z 669.1 (MH) +. Step 2: Synthesis of (R) -3- (trans-4- (aminomethyl) cyclohexanocarboxamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carbo-xyl.
[0209] [00209] To a solution of 3 - ((2R) -2- (trans-4 - ((tert-butoxycarbonyl-amino) methyl) cyclohexanecarboxamido) -2- (2,9,9-trimethyl-3,5 -dioxa-4-bora-tricycle [6,1,1,026] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate from step 1 (200 mg, 0.30 mmol) in anhydrous CH2Cl2 (5 mL) at -78 ° C BCl3 (2.1 mL, 1M in DCM, 2.1 mmol) was added, and the reaction mixture was stirred at the same temperature for 1 h, at which time the reaction mixture was heated to 0 ° C and stirred at the same temperature for an additional 1 h. The reaction was quenched by the addition of water (5 ml) to 0 ° C. The crude product was purified by preparative reverse phase HPLC and dried using lyophilization to provide the product (30 mg) as a white solid. ESI-MS m / z 347 (MH) +. EXAMPLE 2: (R) -3- (trans-4-aminocyclohexanecarboxamido) -2-hydroxy-3.4-dihydro-2H-benzofe1f1.21oxaborinin-8-carboxylic acid
[0210] [00210] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6.1, 1.02.6] dec-4-ylmethyl) -benzoic acid and tran-4- (tert-butoxycarbonylamino) cyclohexanecarboxylic acid after the procedure described in step 1 of Example 1. The crude product was purified by flash chromatography on silica gel (hexane / EtOAc, 2: 1 to 1: 2). ESI-MS m / z 655.1 (MH) +. Step 2: Synthesis of (R) -3- (trans-4-aminocyclohexanecarboxa-mido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8 -carboxylic.
[0211] [00211] Prepared from 3 - ((2R) -2- (trans-4- (tert-butoxycarbo-nylamino) cyclohexanecarboxamido) -2- (2,9,9-trimethyl-3,5-dioxa- Tert -butyl 4-bora-tri-cycle [6,1,1,02,6] dec-4-yl) ethyl) -2-methoxybenzoate and BCl3 after the procedure described in Step 2 of Example 1. The crude product it was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 333 (MH) +. EXAMPLE 3: (R) -3- (2-trans-4- (aminomethyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0212] [00212] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,02'6] dec-4-ylmethyl) -benzoic acid and 2- (trans-4 - ((tert-butoxycarbonylamino) methyl) cyclohexyl) a-skeptic acid after the procedure described in step 1 of Example 1. The crude product was purified by silica gel scintillation chromatography (hexane / EtOAc, 2: 1 to 1: 2). ESI-MS m / z 683.1 (MH) +. Step 2: Synthesis of (R) -3- (2-trans-4- (aminomethyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1, 2] oxaborinin-8-carboxylic.
[0213] [00213] Prepared from 3 - ((2R) -2- (2-trans-4 - (((tert-butoxycarbonylamino) methyl) cyclohexyl) acetamido) -2- (2,9,9-trimethyl -3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate and BCl3 after the procedure described in Step 2 of the Example 1. The crude product was purified by preparative reversed-phase HPLC and dried using lyophilization. ESI-MS m / z 361 (MH) +. EXAMPLE 4: (R) -3- (2- (trans-4- (guanidinomethyl) cyclohexyl) acetamido) -2-hydroxy-3.4-dihydro-2H-benzo [e] [1.21oxaborinin-8- carboxylic
[0214] [00214] Synthesis of (R) -3- (2- (trans-4- (guanidinomethyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] acid [1] , 2] oxaborinin-8-carboxylic.
[0215] [00215] To (R) -3- (2-trans-4- (aminomethyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2 ] oxaborinin-8-carboxylic acid from Example 3 (12 mg) in MeOH (2 ml) was added (1H-pyrazol-1-yl) tert-butyl methanediylidenodicarbamate (12 mg) and stirred for 4 h. The solvent was removed in vacuo. The residue was dissolved in 4 N HCl in dioxane (2 ml) and stirred for 2 h. The solvent was removed in vacuo and the crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 403 (MH) +. EXAMPLE 5: (R) -3- (2- (trans-4 - ((2- (dimethylamino) acetamido) methyl) cyclohexyl) acetamido) -2-hydroxy-3.4-dihydro-2H-benzo [ e1 [1.21oxaborinin-8-carboxylic
[0216] [00216] Ao 3 - ((2R) -2- (2- (trans-4 - ((tert-butoxycarbonylamino) methyl) cyclohexyl) acetamido) -2- (2.9.9-trimethyl-3.5-dioxa -4-bora-tricycle [6.1.1.02,6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate (422 mg. 0.75 mmol from Example 3. Step 1) in a flask 4N HCl was added in dioxane (3 ml) and the reaction mixture was stirred at RT for 1 h. Removal of the solvents gave the product as a yellow foam. Step 2: Synthesis of acid 3 - ((2R) -2- (2- (trans-4 - ((2- (dimethylamino) acetamido) methyl) cyclohexyl) acetamido) -2- (2.9.9-trimethyl- 3.5-dioxa-4-bora-tricycle [6.1.1.02 6] dec-4-yl 1) ethyl) -2-methoxybenzoic.
[0217] [00217] To acid 3 - ((2R) -2- (2- (trans-4- (aminomethyl) cyclohexyl) acetamide) -2- (2.9.9-trimethyl-3.5-dioxa-4-bora -tricycle [6.1.1.026] dec-4-yl) ethyl) -2-methoxybenzoic from step 1 in THF (5 ml). TEA (0.35 ml) was added. followed by 2-bromoacetyl bromide (0.07 mL. 0.8 mmol). The reaction mixture was stirred at RT for 1 h. Water was added and the aqueous phase extracted with EtOAc. The organic phase was dried and concentrated to provide the crude product. which was dissolved in THF (5 ml) and dimethyl amine (1 ml. 2 N in THF) was added. After stirring at RT for 8 h. the volatile components were removed in vacuo and the residue was taken to the next step without further purification. ESI-MS m / z 612.1 (MH) +. Step 3: Synthesis of (R) -3- (2- (trans-4 - ((2- (dimethylamino) acetami-do) methyl) cyclohexyl) acetamido) -2-hydroxy-3,4-di- hydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0218] [00218] Prepared from 3 - ((2R) -2- (2- (trans-4 - ((2- (dimethylamino) acetamido) methyl) cyclohexyl) acetamido) -2- (2, 9,9-trimethyl-3,5-di-oxa-4-bora-tricycle [6,1,1,02'6] dec-4-yl) ethyl) -2-methoxybenzoic and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reversed-phase HPLC and dried using lyophilization. ESI-MS m / z 446 (MH) +. EXAMPLE 6: (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3.4-dihydro-2H-benzo [e1 [1.21oxaborinin-8-carboxylic acid
[0219] [00219] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,02,6] dec-4-ylmethyl) -benzoic acid and 2- (trans-4- (tert-butoxycarbonylamino) cyclohexyl) acetic acid after the procedure described in step 1 of Example 1. The crude product was purified by flash chromatography on silica- gel (hexane / EtOAc, 2: 1 to 1: 2). ESI-MS m / z 613.1 (MH) +. Step 2: Synthesis of (R) -3- (2- (trans-4-aminocyclohexyl) acetami-do) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1, 2] oxaborinin-8-carboxylic.
[0220] [00220] Prepared from 3 - ((2R) -2- (2- (trans-4- (tert-butoxycarbonylamino) cyclohexyl) acetamido) -2- (2,9,9-trimethyl-3, 5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-methoxybenzoic and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 347 (MH) +. EXAMPLE 7: (R) -3- (2- (cis-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0221] [00221] Prepared from the tert-butyl ester of 2-methoxy-3- (2.9.9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec- 4-ylmethyl) -benzoic acid and 2- (cis-4- (tert-butoxycarbonylamino) cyclohexyl) acetic acid after the procedure described in step 1 of Example 1. The crude product was purified by flash chromatography on silica- gel (Hexane / EtOAc. 2: 1 to 1: 2). ESI-MS m / z 613.1 (MH) +. Step 2: Synthesis of (R) -3- (2- (cis-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1.21oxaborinin-8 -carboxylic.
[0222] [00222] Prepared from 3 - ((2R) -2- (2- (cis-4- (tert-butoxycarbonylamino) cyclohexyl) acetamido) -2- (2.9.9-trimethyl-3, 5-dioxa-4-bora-tricycle [6,1,1,02,61dec-4-yl) ethyl) -2-methoxybenzoic and BCb after the procedure described in Step 2 of Example 1. The crude product was purified by HPLC preparative phase and dried using lyophilization. ESI-MS m / z 347 (MH) +. EXAMPLE 7: (R) -3- (2- (trans-4 - (((dimethylamino) methyl) cyclohexyl) -N-methylacetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [ e] [1.21 oxaborinin-8-carboxylic
[0223] [00223] Benzyl chloroformate (1.5 mL, 10.5 mmol) and sodium hydroxide (12.5 mL, 1N, 12.5 mmol) were added to a solution of trans- (4-aminomethylcyclohexyl acid) acetic (2.08 g, 10.0 mmol) in THF (32 mL) and H2O (16 mL). The reaction was stirred at RT for 17 h. The reaction was quenched with 1N HCl and extracted with EtOAc (2x). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated to produce 1.91 g (62%) of product which was carried on to the next step without further purification. ESI-MS m / z 306 (MH) +. Step 2: 3- [2- {2- [4- (Benzyloxycarbonylamino-methyl) -cyclohexyl] -acetylamino} -2- (2,9,9-trimethyl-3,5) tert-butyl ester -dioxa-4-borora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy-benzoic.
[0224] [00224] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,02,6] dec-4-ylmethyl) -benzoic acid and [4- (Benzyloxycarbonylamino-methyl) -cyclohexyl] -acetic acid after the procedure described in step 1 of Example 1. The crude product was purified by flashing silica gel chromatography ( 10 to 100% EtOAc / hexane) ESI-MS m / z 717 (MH) +. Step 3: 3- [2- [2- (4-aminomethyl-cyclohexyl) -acetylamino] -2- (2,9,9-trimethyl-3,5-dioxa-4-bora) tert-butyl ester -tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy-benzoic.
[0225] [00225] A solution of tert-butyl ester of 3- [2- {2- [4- (Benzyloxycarbonylamino-methyl) -cyclohexyl] -acetylamino} -2- (2,9,9-trimethyl-3, 5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy-benzoic (1.56 g, 2.18 mmol) in MeOH (22 mL) was purged with Argon for 5 minutes. Carbon palladium (10%, 0.153 g) was added, the flask evacuated, and the reaction stirred under a hydrogen atmosphere for 6.5 h. The reaction was filtered through a plugged Celite filter frit, washed with MeOH and DCM, and concentrated to provide 1.29 g of crude product which was carried on to the next step without further purification. ESI-MS m / z 583 (MH) +. Step 4: 3- [2 - {[2- (4-methylaminomethyl-cyclohexyl) -acetyl1-methyl-amino} -2- tert-butyl ester -2- (2,9,9-trimethyl-3,5- dioxa-4-bora-tricyclo- [6,1,1,02,6] dec-4-yl) -ethyl1-2-methoxy-benzoic.
[0226] [00226] To a solution of tert-butyl ester of 3- [2- [2- (4-aminomethyl-cyclohexyl) -acetylamino1-2- (2,9,9-trimethyl-3,5-dioxa- DIEA-4-bora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl1-2-methoxy-benzoic (0.208 g, 0.357 mmol) in DCM (3.8 mL) under Argon (0.18 mL, 1.03 mmol) and iodomethane (0.068 mL, 1.09 mmol). The reaction was stirred at RT for 4 h. The reaction was quenched with MeOH, concentrated, and carried on to the next step without further purification. ESI-MS m / z 611 (MH) +. Step 5: (R) -3- (2- (trans-4 - (((dimethylamino) methyl) cyclohexyl) -N-methylacetamido) -2-hydroxy-3,4-dihydro-2H-benzo [ e] [1,2] oxaborinin-8-carboxylic
[0227] [00227] Prepared from 3- [2 - {[2- (4-dimethylaminomethyl-cyclohexyl) -acetyl1-methyl-amino} -2- (2,9,9-trimethyl- 3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy-benzoyl and BCl3 after the procedure described in step 2 of Example 1 The crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 403 (MH) +. EXAMPLE 8: (R) -3- (2- (trans-4-guanidinocyclohexyl) acetamido) - 2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin- 8-carboxylic
[0228] [00228] Synthesis of (R) -3- (2- (trans-4-guanidinocyclohexyl) ace-tamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1, 2] oxaborinin-8-carboxylic
[0229] [00229] Prepared from 3 - ((R) -2- (2- (trans-4-aminocyclohexyl) acetamido) -2-boronoethyl) -2-hydroxybenzoic acid (Example 6) after the procedure described in Example 4. ESI-MS m / z 389 (MH) +. EXAMPLE 9: (R) -3- (2- (trans-4 - (((2-aminoethylamino) methyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e ] [1,2] oxaborinin-8-carboxylic
[0230] [00230] A sealable reaction tube was charged with 3- [2- [2- (4-aminomethyl-cyclohexyl) -acetylamino] -2- (2.9.9-trimethyl-3,5) tert-butyl ester -dioxa-4-bora-tricycle [6,1,1.02,6] dec-4-yl) -ethyl] -2-methoxy-benzoic (0.205 g. 0.352 mmol). potassium carbonate (0.057 g. 0.412 mmol). 2- (boc-amino) ethyl bromide (0.095 g. 0.424 mmol). and DMF (3.0 mL). The tube was sealed and the reaction heated to 65 ° C for 24 h. The reaction was cooled to RT. The reaction was diluted with EtOAc and washed with 5% aqueous LiCl (2x) and brine. The organic layer was dried over Na2SO4. filtered and concentrated to provide 0.120 g of crude product which was carried on to the next step without further purification. ESI-MS m / z 726 (MH) +. Step 2: (R) -3- (2- (trans-4 - (((2-aminoethylamino) methyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e ] [1.2] oxaborinin-8-carboxylic
[0231] [00231] Prepared from 3- [2- (2- {4 - [(2-tert-butoxycarbonylamino-ethylamino) -methyl] -cyclohexyl} -acetylamino acid) tert-butyl ester - 2- (2.9 .9-trimethyl-3,5-dioxa-4-bora-tricycle [6.1.1.02.6] dec-4-yl) -ethyl] -2-methoxy-benzoic and BCl3 after the procedure described in Step 2 of Example 1 The crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 404 (MH) +. EXAMPLE 10: (3R) -3- (2- (4- (aminomethyl) cyclohexylidene) acetamide) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1.21oxaborinin- 8-carboxylic
[0232] [00232] Potassium tert-butoxide (0.479 g. 4.27 mmol) was added to a solution of triethylphosphonoacetate (0.85 mL. 4.28 mmol) in DMF (6.5 mL) under Argon and the reaction stirred at RT for 10 min. (4-Oxo-cyclohexylmethyl) -carbamic acid tert-butyl ester (0.643 g. 2.83 mmol) in DMF (6.5 mL) was added dropwise for 8 min. After 20 min of shaking. a precipitate was observed and additional DMF (6.5 ml) was added and the reaction stirred for an additional 17 h. The reaction was poured into ice-cold H2O and extracted with Et2O (3x). The combined organic layers were washed with H2O and brine. dried by Na2SO4. filtered. and concentrated. Scintillation chromatography (0 to 30% EtOAc / hexane) gave 0.784 g (93%) of product. ESI-MS m / z 298 (MH) +. Step 2: [4- (tert-butoxycarbonylamino-methyl) -cyclohexylidene] -acetic acid.
[0233] [00233] To a solution of [4- (tert-Butoxycarbonylamino-methyl) -cyclohexylidene] -acetic acid ethyl ester (0.518 g. 1.74 mmol) in MeOH (16 mL) and THF (4 mL) Sodium hydroxide (9.0 mL, 1N. 9.0 mmol) was added and the reaction stirred at RT for 23 h. The reaction was quenched with H2O and extracted with EtOAc (2x). The aqueous layer was acidified to pH ~ 1 with 1 N HCl and extracted with EtOAc (3x). The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo to provide 0.300 g (64%) of the product. ESI-MS m / z 270 (MH) +. Step 3: 3- [2- {2- [4- (tert-butoxycarbonyl-mino-methyl) -cyclohexylidene1-acetylamino} -2- (2,9,9-trimethyl-3, tert-butyl ester) 5-dioxa-4-bora-tricycle [6,1,1,02,61dec-4-yl) -ethyl1-2-methoxy-benzoic.
[0234] [00234] Prepared from 2-methoxy-3- [2- (2.9.9-trimethyl-3,5-dioxa-4-bora-tricycle] tert-butyl ester] [6.1.1.02.61dec-4-yl ) -2- (trime-tilsilanil-mino) -ethyl] -benzoic acid [4- (tert-butoxycarbonylamino-methyl) -cyclohexylidene] -acetic after the procedure described in Step 1 of Example 1. The crude product was purified by silica gel scintillation chromatography (0 to 100% EtOAc / hexane). ESI-MS m / z 681 (MH) +. Step 4: (3R) -3- (2- (4- (aminomethyl) cyclohexylidene) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin -8-carboxylic
[0235] [00235] Prepared from 3- [2- {2- [4- (tert-butoxycarbonylamino-methyl) -cyclohexylidene] -acetylamino} -2- (2,9,9-trimethyl) tert-butyl ester -3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy-benzoic and BCl3 after the procedure described in Step 2 of Example 1 The crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 359 (MH) +. EXAMPLE 11: (R) -3- (2- (4- (aminomethyl) -1- (nitromethyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1.21oxaborinin-8-carboxylic
[0236] [00236] A sealable reaction tube was loaded with 2,8,9-T risisobutyl-2,5,8,9-tetraaza-1-phosfabicyclo [3,3,3] undecane (0.16 mL, 0.450 mmol) and THF (1.0 mL) under Argon. Nitromethane (0.23 mL, 4.24 mmol) was added and the reaction stirred at room temperature for 5 min, then cooled to 0 ° C for 15 min. [4- (tert-Butoxycarbonylamino-methyl) -cyclohexylidene] -acetic acid ethyl ester (0.261 g, 0.878 mmol) in THF (1.5 mL) was added slowly and the reaction stirred at RT for 15 min. The tube was sealed and the reaction heated to 70 ° C for 17 h. The reaction was cooled to room temperature, cooled quickly with 0.5M HCl, and extracted with EtOAc (2x). The combined organic layers were washed with saturated NaHCO3 and brine, dried over Na2SO4, filtered, and concentrated to provide 0.304 g of crude product which was carried on to the next step without further purification. ESI-MS m / z 359 (MH) +. Step 2: [4- (tert-butoxycarbonylamino-methyl) -1-nitromethyl-cyclohexyl] -acetic acid.
[0237] [00237] To a solution of [4- (tert-butoxy-bonylamino-methyl) -1-nitromethyl-cyclohexyl] -acetic acid ethyl ester (0.304 g, 0.848 mmol) in MeOH (5.0 mL) and THF (1.5 ml) was added sodium hydroxide (4.2 ml, 1N, 4.2 mmol) and the reaction stirred at RT for 5 h. The reaction was acidified to pH ~ 2 with 1N HCl and extracted with EtOAc (3x). The combined organic layers were dried over Na2SO4, filtered, and concentrated to provide 0.308 g of crude product which was carried on to the next step without further purification. ESI-MS m / z 331 (MH) +. Step 3: 3- [2- {2- [4- (tert-butoxycarbonyl-mino-methyl) -1-nitromethyl-cyclohexyl] -acetylamino} -2- tert-butyl ester} -2- (2,9,9 -trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy-benzoic.
[0238] [00238] Prepared from 2-methoxy-3- [2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle] tert-butyl ester] [6,1,1,02 , 6] dec-4-yl) -2- (trime-tilsilanil-amino) -ethyl] -benzoic acid and [4- (tert-butoxycarbonylamino-methyl) -1-nitromethyl-cyclohexyl] -acetic after procedure described in Step 1 of Example 1. The crude product was purified by silica gel scintillation chromatography (0 - 100% EtOAc / hexane). ESI-MS m / z 742 (MH) +. Step 4: (R) -3- (2- (4- (aminomethyl) -1- (nitromethyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [ and] [1,2] oxaborinin-8-carboxylic.
[0239] [00239] Prepared from 3- [2- {2- [4- (tert-butoxycarbonylamino-methyl) -1-nitromethyl-cyclohexyl] -acetylamino} -2- tert-butyl ester -2- (2.9 , 9-trimethyl-3,5-dioxa-4-bora-tricycle [6.1, 1.02.6] dec-4-yl) -ethyl] -2-methoxy-benzoic and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reversed-phase HPLC and dried using lyophilization. ESI-MS m / z 420 (MH) +. EXAMPLE 12: (R) -3- (2- (trans-4 - ((S) -2.3-diaminopropanamido) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [ e] [1,2] oxaborini-na-8-carboxylic
[0240] [00240] Synthesis of 3 - ((R) -2-borono-2- (2-trans-4 - ((S) -2.3-diaminopropanamido) cyclohexyl) acetamido) ethyl) -2-hydroxybenzoic acid.
[0241] [00241] To (S) -2- (benzyloxycarbonylamino) -3- (tert-butoxycarbonylamino) propanoic acid (85 mg. 0.25 mol) in DCM / DMF (4 mL, 1/1) was added NMM ( 1.2 eq) followed by HATU (95 mg. 0.26 mol). The reaction mixture was stirred at RT for 1 h. In a separate bottle. 3 - ((2R) -2- (2- (trans-4- (tert-butoxycarbonylamino) cyclohexyl) acetamido) -2- (2.9.9-trimethyl-3,5-dioxa- 4-bora-tricycle [6.1.1.02.6] dec-4-yl) ethyl) -2-methoxybenzoic (133 mg. 0.2 mmol Example 6) was treated with 4 N HCl in dioxane (2 mL) and the resulting solution was stirred at RT for 2 h. The solvent was then removed under reduced pressure. The above active ester preparation was added to this residue and the reaction mixture was stirred overnight at RT. Water was added and the aqueous phase extracted with EtOAc. The organic phase was washed with 1N HCl, sat. NaHCO3, brine, dried and concentrated in vacuo to provide the product (0.100 g) as brown oil without further purification. ESI-MS m / z 833.1 (MH) +. The residue was then dissolved in MeOH and Pd in carbon cat. (20 mg) was added and stirred under a hydrogen atmosphere overnight. After filtration and removal of the solvent, the residue was treated with BCl3 after the procedure described in step 2 of Example 1 to provide the title compound. ESI-MS m / z 433 (MH) +. EXAMPLE 13: (R) -3- (2- (trans-4- (dimethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2 ] oxaborinin-8-carboxylic
[0242] [00242] Synthesis of (R) -3- (2- (trans-4- (dimethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] acid [1] , 2] oxaborinin-8-carbo-xyl
[0243] [00243] To (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin -8-carboxylic (10.0 mg from Example 6) in MeOH (5 ml) was added formaldehyde (1.0 ml, 37% solution), followed by 10% Pd / C (20 mg). The reaction mixture was hydrogenated in an H2 balloon for 3 h. The reaction mixture was filtered and the solvent was removed in vacuo. The final product was purified by preparative reversed-phase HPLC and dried using lyophilization. ESI-MS m / z 433 (MH) +. EXAMPLE 14: (R) -3- (2- (trans-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] acid [1] , 2] oxaborinin-8-carboxylic
[0244] [00244] To acid (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin -8-carboxylic (Example 6, 15 mg) in MeOH (2 ml) tert-butyl 2-oxoethylcarbamate (20 mg) was added. Pd / C (10% by weight, 10 mg) was added and the reaction mixture was stirred under H2 balloon overnight. The reaction mixture was filtered and the solvent was then removed under reduced pressure and the residue was carried on to the next step without further purification. ESI-MS m / z 490.1 (MH) +. Step 2: Synthesis of (R) -3- (2- (trans-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0245] [00245] To (R) -3- (2- (trans-4- (2- (tert-butoxycarbonylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro- 2H-benzo [e] [1,2] oxaborinin-8-carboxylic (20 mg) in a flask was added 1 ml of 4 N HCl in dioxane. The resulting reaction mixture was stirred at RT for 2 h. The solvent was removed in vacuo and the residue was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 390 (MH) +. EXAMPLE 15: (R) -2-hydroxy-3- (2- (trans-4- (piperazin-1-yl) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0246] [00246] To 2- (4-oxocyclohexyl) acetic acid (0.576 g, 3.7 mmol) and tert-butyl piperazine-1-carboxylate (0.700 g, 3.7 mmol) in MeOH, Pd was added in carbon (80 mg), and the resulting reaction mixture was stirred under a hydrogen atmosphere overnight. The catalyst was removed by filtration and the solvent was removed under reduced pressure to provide the acid as a white solid (1.2 g, 99%). Step 2: Synthesis of 4- (4- {3-tert-butoxycarbonyl-2-methoxy-phenyl) -1- (2,9,9-trimethyl-3,5-dioxa-4-tert-butyl ester) -bora-tricycle [6,1,1,02,6] dec-4-yl) -ethylcarbamoyl] -methyl} -cyclohexyl) -piperazine-carboxylic.
[0247] [00247] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,0 26] dec -4-ylmethyl) -benzoic acid and 2- (4- (4- (tert-butoxycarbonyl) piperazin-1-yl) cyclohexyl) acetic acid after the procedure described in step 1 of Example 1. The crude product was purified by silica gel scintillation chromatography (hexane / EtOAc, 2: 1 to 1: 2). ESI-MS m / z 638.1 (MH) +. Step 3: Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (piperazin-1-yl) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [ and] [1,2] oxaborinin-8-carboxylic acid.
[0248] [00248] Prepared from 4- (4- {3-tert-butoxycarbonyl-2-methoxy-phenyl) -1- (2,9,9-trimethyl-3,5-dioxa-4) tert-butyl ester -bora-tricycle [6,1,1,02,6] dec-4-yl) -ethylcarbamoyl] -methyl} -cyclohexyl) -piperazine-carboxylic and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reverse phase HPLC to obtain the trans isomer as the main product, which was dried using lyophilization. ESI-MS m / z 416 (MH) +. EXAMPLE 16: (R) -2-hydroxy-3- (2- (cis-4- (piperazin-1-yl) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0249] [00249] Synthesis of (R) -2-hydroxy-3- (2- (cis-4- (piperazin-1-yl) cyclohexyl) acetamido) -3,4-dihydro-2H- benzo [e] [1,2] oxaborinin-8-carbo-xyl.
[0250] [00250] Prepared from 4- (4- {3-tert-butoxycarbonyl-2-methoxy-phenyl) -1- (2,9,9-trimethyl-3,5-dioxa-4) tert-butyl ester -bora-tricycle [6,1,1,02,6] dec-4-yl) -ethylcarbamoyl] -methyl} -cyclohexyl) -piperazine-carboxylic (Example 16, step 2) and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reversed-phase HPLC to obtain the cis isomer as the by-product, which was dried using lyophilization. ESI-MS m / z 416 (MH) +. EXAMPLE 17: (3R) -3 - [[2 - [(1S, 3S, 4S) -3,4-bis (aminomethyl) cyclohexyl] acetyl] amino] -2-hydroxy-3,4-di- hydro-1,2-benzoxaborinin-8-carbo-xylic acid (3R) -3 - [[2 - [(1R, 3R, 4R) -3,4-bis (aminomethyl) cyclohexyl] acetyl] amino] -2-hydroxy-3,4-dihydro-1,2-benzoxaborinin-8-carboxylic
[0251] [00251] To a cooled (-5 ° C) solution of N, N-dibenzylamine (19.3 ml, 100 mmol) in DCM (200 ml) is added TEA (13.8 ml, 100 mmol). To this solution is added, dripping for 5 minutes, fumarolyl chloride (4.2 mL, 40 mmol). Upon complete addition, the cold bath is removed and stirring continued for 1 h. This solution is diluted with DCM, washed with HCl (approx. 100 ml of 1M aq.) Then water (2x), dried over MgSO4 and concentrated. The solid residue is crystallized from hot toluene (109 ° C) to provide the title compound (17.9g) as a white solid. ESI-MS (m / z) 475 (MH) +. Step 2: Synthesis of (trans) -N, N, N ', N'-tetrabenzyl-4-oxo-cyclohexane-1,2-dicarboxamide.
[0252] [00252] To a suspension of (E) -N, N, N ', N'-tetrabenzylbut-2-ene-diamide (10.6 g, 22 mmol) in o-xylene (106 mL) is added hydroquinone (180 mg, 1.63 mmol) followed by 2-trimethylsilyloxy-1,3-butadiene (9.48 g, 66 mmol). The resulting mixture is sprayed with Argon and then sealed. The mixture is then heated to 135 ° C and stirred at this temperature for 87 h. The resulting solution is cooled, then concentrated under reduced pressure. The residue is purified by chromatography on silica (240 g of silica elution with 10% ethyl acetate in hexane) to provide 12.81 g of product as an oil. This crude product is absorbed in THF (10 ml) and MeOH (40 ml). To this solution is added potassium carbonate (2.76g, 20 mmol). The resulting mixture is stirred for 10 min then diluted with Et2O, washed with water and brine, dried over MgSO4 and concentrated. The residue is purified by chromatography on silica (160 g elution of silica with 20% ethyl acetate / 20% dichloromethane in hexane) to provide the title compound (10.11 g) as an oil. ESI-MS (m / z) 545 (MH) +. Step 3: Synthesis of ethyl 2 - [(trans) -3,4-bis (dibenzylcarbamoyl) cyclohexyl-dene] acetate.
[0253] [00253] To a cooled suspension (-5 ° C) of sodium hydride (846 mg, 60% dispersion in mineral oil, 21.2 mmol) in THF (60 mL) is added, by dripping, triethylphosphonoacetate (4.2 mL, 21.2 mmol). Upon complete addition, the cold bath is removed and stirring continued for 20 min. The resulting solution is cooled to -5 ° C. To this solution is added a solution of (trans) -N, N, N ', N-tetrabenzyl-4-oxo-cyclohexane-1,2-dicarboxamide (10.11g, 18.4 mmol) in THF (10 mL). Upon complete addition, the cold bath is removed and stirring continued for 30 min. To this solution is added HCl (30 ml, 1M aqueous). The resulting mixture is diluted with Et2O, washed with brine, dried over MgSO4 and concentrated. The residue is purified by chromatography on silica (160g elution of silica with 20% ethyl acetate / 10% dichloromethane in hexane) to provide the title compound (9.89 g) as an oil (4: 1 mixture of Z isomers) . ESI-MS (m / z) 615 (MH) +. Step 4: Synthesis of 2 - [(1R, 3R, 4R) -3,4-bis (dibenzylcarbamoyl) cyclohexyl] (racemic) -methyl acetate.
[0254] [00254] To a solution of 2 - [(trans) -3,4-bis (dibenzylcarbamoyl) cyclohexylidene] ethyl acetate (9.27 g, 15 mmol) in dry MeOH (75 mL) is added Magnesium tape (1.08 g, 45 mmol). The suspension was stirred for 5 h. To this homogeneous solution, another batch of magnesium tape (200 mg, 8.2 mmol) is added. This mixture is stirred for 17 h. The resulting solution is extracted with EtOAc, washed with brine, dried over MgSO4 and concentrated. The residue is purified by chromatography on silica (eluting with 20% ethyl acetate / 10% dichloromethane in hexane) to provide the title compound (8.27 g) as a foam. ESI-MS (m / z) 617 (MH) +. Step 5: Synthesis of (racemic) -2 - [(1R, 3R, 4R) -3,4-bis [(dibenzylamino) methyl] cyclohexyl] ethanol.
[0255] [00255] To a cooled (-10 ° C) solution of 2 - [(1R, 3R, 4R) -3,4-bis (dibenzylcarbamoyl) cyclohexyl] (racemic) -methyl acetate (8.18 g, 13.5 mmol) in THF (40 mL) is added dropwise with lithium aluminum hydride (50 mL, 1 M in THF). Upon complete addition, the cold bath is removed and stirring continued for 15 min. The resulting solution is heated to 55 ° C and stirred at this temperature for 2.5 h. This solution is cooled to 0 ° C. To the resulting solution, water (1.9 ml) is added dropwise then NaOH (1.9 ml, 5M aqueous) then water (0.95 ml). The resulting mixture is diluted with Et2O (100 ml), filtered through Celite and the filtrate concentrated in vacuo to provide the title compound as an oil (8.1 g). ESI-MS (m / z) 547 (MH) +. Step 6: Synthesis of (racemic) -tert-butyl N - [[(1S, 2S, 4S) -2 - [(tert-butoxycarbonylamino) methyl] -4- (2-hydroxyethyl) cyclohexyl] methyl] carbamate .
[0256] [00256] To a mixture of (racemic) -2 - [(1R, 3R, 4R) -3,4-bis [(diben-zylamino) methyl] cyclohexyl] ethanol (8.1g, 13 mmol) and hydroxide of palladium on carbon (1.0 g, 20% palladium by weight) is added MeOH (70 ml). The resulting mixture is flushed with hydrogen gas (1 atm) and stirred for 17 h. The system is then flushed with Argon, diluted with DCM and filtered through Celite. The filtrate is concentrated under vacuum. The residue is absorbed in DCM (30 ml) and THF (20 ml). To this solution is added DIEA (4.6 mL, 26 mmol) followed by di-tert-butyl dicarbonate (5.66 g, 26 mmol). The resulting solution is stirred for 2 h, then diluted with EtOAc, washed with water and brine, dried over MgSO4 and concentrated. The residue is purified by chromatography on silica (160g silica, eluting with 80% ethyl acetate in hexane) to provide the title compound (3.1 g) as a viscous oil. 1H NMR (CDCl3) δ 0.7 - 1.0 (m, 1H), 1.05 - 1.35 (m, 4H), 1.4 - 1.75 (m, 4H), 1.46 (s , 18H), 1.75 - 1.96 (m, 2H), 2.9 - 3.05 (m, 2H), 3.05 - 3.75 (bm, 5H), 4.6 - 4.8 (bd, 1H), 4.95 (bs, 1H). Step 7: Synthesis of (racemic) -2 - [(1S, 3S, 4S) -3,4-bis [(tert-butoxycarbonylamino) methyl] cyclohexyl] acetic acid.
[0257] [00257] To a solution of N - [[(1S, 2S, 4S) -2 - [(tert-butoxycarbonyl-mino) methyl] -4- (2-hydroxyethyl) cyclohexyl] methyl] carbamate of (racemic) -tert-butyl (772mg, 2mmol) in acetonitrile (3ml), carbon tetrachloride (3ml) and water (4.5ml) is added sodium periodate (1.28g, 6mmol) followed by RuCl3. H2O (21 mg, 0.1 mmol). The resulting mixture is stirred for 2 h, then diluted with EtOAc, washed with brine, dried over MgSO4 and concentrated. The residue is taken up in a saturated sodium carbonate solution and extracted with EtOAc. The aqueous phase is acidified with HCl (2M aqueous) and extracted with EtOAc. The organic extract is washed with brine, dried over MgSO4 and concentrated. This residue is purified by chromatography on silica (eluting with ethyl acetate) to provide the title compound (394 mg) as a viscous oil. 1H NMR (DMSO-d6) δ 0.7 - 1.0 (m, 1H), 1.0 - 1.9 (m, 9H), 1.39 (s, 18H), 2.0 - 2.3 (m, 2H), 2.6 - 3.18 (m, 4H), 6.6 - 6.8 (bm, 2H). Step 8: Synthesis of 3 - ((2R) -2 - (- [(1S, 3S, 4S) -3,4-bis [(tert-butoxycarbon-nylamino) methyl] cyclohexyl] acetamido) -2- ( 2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate and 3- ( (2R) - 2- (- [(1R, 3R, 4R) -3,4-bis [(tert-butoxycarbonylamino) methyl] cyclohexyl] acetamido) -2- (2,9,9-trimethyl-3, Tert-butyl 5-dioxa-4-bora-tricycle [6,1,1,026] dec-4-yl) ethyl) -2-methoxybenzoate.
[0258] [00258] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,0 2,6 ] dec-4-ylmethyl) -benzoic acid and (racemic) -2 - [(1S, 3S, 4S) -3,4-bis [(tert-butoxycarbonyl-mino) methyl] cyclohexyl] acetic acid after the procedure described in Step 1 of Example 1. Unless the crude product is purified by chromatography on silica (elution with 30% ethyl acetate in hexane then 60% ethyl acetate in hexane) to provide the title compound ESI- MS (m / z) 834 (MNa) +. Step 9: Synthesis of (3R) -3 - [[2 - [(1S, 3S, 4S) -3,4-bis (aminomethyl) cyclohexyl] acetyl] amino] -2-hydroxy-3,4- dihydro-1,2-benzoxaborinin-8-carboxylic acid (3R) -3 - [[2 - [(1R, 3R, 4R) -3,4-bis (aminomethyl) cyclohexyl] acetyl] amino] -2-hydroxy-3,4-dihydro-1,2-benzoxaborinin-8-carboxylic.
[0259] [00259] Prepared from 3 - ((2R) -2 - (- [(1S, 3S, 4S) -3,4-bis [(tert-bu-toxicarbonylamino) methyl] cyclohexyl] acetamido) -2 - tert-butyl (2,9,9-trimethyl-3,5-dio-xa-4-bora-tricycle [6,1,1,02'6] dec-4-yl) ethyl) -2-methoxybenzoate and 3 - ((2R) -2 - (- [(1 R, 3R, 4R) -3,4-bis [(tert-butoxycarbonylamino) methyl] cyclohexyl] acetamido) -2- (2,9,9 -trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate after the procedure described in Step 2 of the Example 1. ESI-MS (m / z) 390 (MH) +. EXAMPLE 18: (3R) -3 - [[((3S, 4S) -3,4-diaminocyclopentanocarbonyl] amino] -2-hydroxy-3,4-dihydro-1,2-benzoxaborinin-8-carboxylic acid (3R) -3 - [[(3R, 4R) -3,4-diaminocyclopentanocarbonyl] amino] -2-hydroxy-3,4-dihydro-1,2-benzoxaborinin-8-carboxylic
[0260] [00260] To a cooled suspension (-20 ° C) of manganese triacetate (8.02 g, 30 mmol) in acetonitrile (120 mL) is added sodium azide (3.4 g, 50 mmol). To this mixture, a solution of ethyl cyclopent-3-ene-1-carboxylate (1.4 g, 10 mmol) in TFA (14 mL) is added by dripping over approximately 10 min. The resulting mixture is stirred at -25 ° C to -19 ° C for 3 h, then allowed to warm to RT and stirred for 17 h. To this mixture is added sodium thiosulfate (30 mL, 10% aqueous). This mixture is stirred for 5 min then extracted with hexane. The hexane extract is washed with a saturated solution of saturated sodium bicarbonate (3x), then brine, dried over MgSO4 and concentrated. This residue is purified by chromatography on silica (90 g silica, eluting with 10% ethyl acetate in hexane) to provide the title compound (1.278 g) as an oil. 1H NMR (DMSO-d6) δ 1.15 (t, J = 7Hz, 3H), 1.68 - 1.91 (m, 2H), 2.13 - 2.35 (m, 2H), 3.01 (m, 1H), 3.97 (m, 2H), 4.05 (q, J = 7Hz, 2H). Step 2: Synthesis of ethyl (racemic) (3S, 4S) -3,4-bis (tert-butoxycarbonyl-mino) cyclopentanecarboxylate.
[0261] [00261] To a solution of (racemic) -ethyl (3S, 4S) -3,4-diazidocyclopentanocarbon-xylate (3.8 g, 16.9 mmol) in THF (85 mL) is added triphenylphosphine (9.7 g, 37.18 mmol). The resulting solution is stirred for 17 h. To this solution is added water (4.2 ml). The resulting solution is stirred for 6 h. To this solution is added DIEA (8.9 mL, 51 mmol) followed by di-tert-butyl-dicarbonate (11.05 g, 51 mmol). This mixture is stirred for 2 h then concentrated in vacuo. The residue is purified by chromatography on silica (230 g silica, eluting with 20% ethyl acetate in hexane) to provide the title compound (3.79 g) as a white solid. 1H NMR (DMSO-d6) δ 1.15 (t, J = 7Hz, 3H), 1.36 (s, 18H), 1.5 - 1.66 (m, 2H), 1.91 - 2.11 (m, 2H), 2.81 (m, 1H), 3.61 (bm, 2H), 4.01 (q, J = 7Hz, 2), 6.8 (bs, 2H). Step 3: Synthesis of (racemic) - (3S, 4S) -3,4-bis (tert-butoxycarbonyl-mino) cyclopentanecarboxylate.
[0262] [00262] To a solution of ethyl (racemic) (3S, 4S) -3,4-bis (tert-butoxy-carbonylamino) cyclopentanecarboxylate (790 mg, 2.12 mmol) in MeOH (3 mL) ml) sodium hydroxide (3 ml, 1M aqueous) is added. The resulting solution is stirred for 20 min then acidified with HCl (1M, aqueous). This mixture is diluted with EtOAc, washed with brine, dried over MgSO4 and concentrated to provide the title compound (671 mg) as a solid. 1H NMR (DMSO-d6) δ 1.38 (s, 18H), 1.46 - 1.62 (m, 2H), 1.87 - 2.11 (m, 2H), 2.77 (m, 1H ), 3.6 (bm, 2H), 6.75 (bs, 2H), 12.15 (bs, 1H). Step 4: Synthesis of 3 - ((2R) -2 - ((3S, 4S) -3,4-bis (tert-butoxycarbonyl-mino) cyclopentanecarboxamido) -2- (2,9,9-trimethyl-3,5 -dioxa-4-bora-trici-clo [6,1,1,026] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate and 3 - ((2R) -2- (3R, 4R) -3 , 4-bis (tert-butoxycarbonylamino) cyclopentanecarboxamido) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6.1, 1.02'6] dec-4-yl ) tert-butyl ethyl) -2-methoxy-benzoate.
[0263] [00263] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,02'6] dec-4-ylmethyl) -benzoic and (racemic) - (3S, 4S) -3,4-bis (tert-butoxycarbonylamino) cyclopen-tanocarboxylate after the procedure described in Step 1 of Example 1. Unless crude product is purified by chromatography on silica (elution with 30% ethyl acetate in hexane then 60% ethyl acetate in hexane) to provide the title compound ESI-MS (m / z) 778 (MNa) +. Step 5: Synthesis of (3R) -3 - [[((3S, 4S) -3,4-diaminocyclopentane-carbonyl] amino] -2-hydroxy-3,4-dihydro-1,2-benzoxaborinin-8 acid -carboxylic acid and (3R) -3 - [[(3R, 4R) -3,4-diaminocyclopentanocarbonyl] amino] -2-hydroxy-3,4-dihydro-1,2-benzoxaborinin-8-carboxylic acid .
[0264] [00264] Prepared from 3 - ((2R) -2 - ((3S, 4S) -3,4-bis (tert-butoxy-carbonylamino) cyclopentanecarboxamido) -2- (2,9,9-trimethyl-3 , 5-dioxa-4-bora-tricycle [6,1,1,02'6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate and 3 - ((2R) -2- (3R, 4R) -3,4-bis (tert-butoxycarbonylamino) cyclopentanecarboxamido) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02'6] dec -4-yl) ethyl) -2-tert-butyl methoxybenzoate after the procedure described in Step 2 of Example 1. ESI-MS (m / z) 334 (MH) +. EXAMPLE 19: (3R) -3 - [[((1S, 3S, 4S) -3,4-diaminocyclohexanocarbonyl] amino] -2-hydroxy-3,4-dihydro-1,2-benzoxaborinin acid -8-carboxylic and (3R) -3 - [[((1R, 3R, 4R) -3,4-diaminocyclohexanecarbonyl] amino] -2-hydroxy-3,4-dihydro-1,2-benzoxaborinin -8-carboxylic
[0265] [00265] Prepared from methyl cyclohex-3-ene-1-carboxylate, using essentially the same procedure described in Step 1 of Example 19. This material was used without purification. Step 2: Synthesis of (racemic) methyl (1S, 3R, 4R) -3,4-bis (tert-butoxycarbonylamino) cyclohexanecarboxylate.
[0266] [00266] Prepared from (racemic) -methyl (3R, 4R) -3,4-diazidocyclohexanocarboxylate using essentially the same procedure described in Step 2 of Example 19. 1H NMR (CDCb) δ 1.3 - 1.45 (m, 3H), 1.40 (s, 18H), 1.91 (m, 1H), 2.10 (m, 1H), 2.41 (m, 1H) m, 2.72 (m, 1H), 3.28 (m, 1H), 3.52 (m, 1H), 3.68 (s, 3H), 4.79 (bs, 1H), 4.93 (bs, 1H) . Step 3: Synthesis of (racemic) - (1S, 3R, 4R) -3,4-bis (tert-butoxy-carbonylamino) cyclohexanecarboxylic acid.
[0267] [00267] Prepared from (racemic) methyl (1S, 3R, 4R) -3,4-bis (tert-butoxycarbo-nylamino) cyclohexanecarboxylate using essentially the same procedure as described in Step 3 of Example 19. 1H NMR (CDCl3) δ 1.28 - 1.62 (m, 3H), 1.42 (s, 18H), 1.86 (m, 1H), 2.20 (m, 1H), 2.47 (m , 1H), 2.75 (m, 1H), 3.32 (m, 1H), 3.53 (m, 1H), 4.91 (bd, 1H), 5.55 (bd, 1H). Step 4: Synthesis of 3 - ((2R) -2 - ((1S, 3R, 4R) -3,4-bis (tert-butoxycarbon-nylamino) cyclohexanecarboxamido) -2- (2,9,9-trimethyl -3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate and 3 - ((2R) - 2- ( (1R, 3S, 4S) -3,4-bis (tert-butoxycarbonylamino) cyclohexanecarboxy-wet) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6, 1,1,02,6] tert-butyl dec-4-yl) ethyl) -2-methoxybenzoate.
[0268] [00268] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,0 2,6 ] dec-4-ylmethyl) -benzoic acid and (racemic) - (1S, 3R, 4R) -3,4-bis (tert-butoxycarbonylamino) cyclohexanecarboxylic acid following the procedure described in Step 1 of Example 1. Unless the crude product is purified by chromatography on silica (elution with 30% ethyl acetate in hexane then 60% ethyl acetate in hexane) to provide the title compound ESI-MS (m / z) 792 (MNa) +. Step 5: Synthesis of (3R) -3 - [[((1S, 3S, 4S) -3,4-diaminocyclohexanecarbonyl] amino] -2-hydroxy-3,4-dihydro-1,2-benzoxaborinin acid -8-carboxylic and (3R) -3 - [[((1R, 3R, 4R) -3,4-diaminocyclohexanocarbon-nil] amino] -2-hydroxy-3,4-dihydro-1,2 -benzoxaborinin-8-carboxylic.
[0269] [00269] Prepared from 3 - ((2R) -2 - ((1S, 3R, 4R) -3,4-bis (tert-bu-toxicarbonylamino) cyclohexanecarboxamido) -2- (2,9,9 -trimethyl-3,5-dioxa -4-bora-tricycle [6,1,1,02'6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate and 3 - ((2R) -2 - (((1R, 3S, 4S) -3,4-bis (tert-butoxycarbonylamino) cyclohexane-carboxamido) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [ 6,1,1,026] dec-4-yl) ethyl) -2-methoxybenzoate tert-butyl after the procedure described in Step 2 of Example 1. ESI-MS (m / z) 348 (MH) +. EXAMPLE 20: (R) -3- (3- (trans-4-aminocyclohexyl) propanamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin- 8-carboxylic Step 1: Synthesis of 3 - ((2R) -2- (3- (trans-4- (tert-butoxycarbonylamino) cyclohexyl) propanamido) -2- (2,9,9 -trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-methoxybenzoic.
[0270] [00270] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,02,6] dec-4-ylmethyl) -benzoic acid and 3- (trans-4- (tert-butoxycarbonylamino) cyclohexyl) propanoic acid after the procedure described in step 1 of Example 1. The crude product was purified by flashing silica chromatography -gel (hexane / EtOAc, 2: 1 to 1: 2). ESI-MS m / z 683.1 (MH) +. Step 2: Synthesis of (R) -3- (3- (trans-4-aminocyclohexyl) propanamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carboxylic.
[0271] [00271] Prepared from 3 - ((2R) -2- (3- (trans-4- (tert-butoxycarbonylamino) cyclohexyl) propanamido) -2- (2,9,9 -trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,026] dec-4-yl) ethyl) -2-methoxybenzoic and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 361 (MH) +. EXAMPLE 21: (R) -3- (3- (trans-4-guanidinocyclohexyl) propanamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin- 8- carboxylic
[0272] [00272] Synthesis of (R) -3- (3- (trans-4-guanidinocyclohexyl) pro-panamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1, 2] oxaborinin-8-carbo-xyl.
[0273] [00273] Prepared from (R) -3- (3- (trans-4-aminocyclohexyl) propanamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1, 2] oxaborin-na-8-carboxylic (Example 21) after the procedure described in Example 4. ESI-MS m / z 403 (MH) +. EXAMPLE 22: (R) -3- (2- (trans-4 - ((2- (dimethylamino) ethyl) (methyl) amino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro acid -2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0274] [00274] Synthesis of (R) -3- (2- (trans-4 - ((2- (dimethylamino) ethyl) (methyl) amino) cyclohexyl) acetamido) -2-hydroxy-3,4-di acid -hydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0275] [00275] Prepared from (R) -3- (2- (trans-4- (2-aminoethyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- acid benzo [e] [1,2] oxaborinin-8-carboxylic (Example 15) after the procedure described in Example 14. ESI-MS m / z 432 (MH) +. EXAMPLE 23: (R) -3- (2- (trans-4- (4-carbamimidoylpiperazin-1-yl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [ e] [1,2] oxaborinin-8-carboxylic
[0276] [00276] Synthesis of (R) -3- (2- (trans-4- (4-carbamimidoylpipera-zin-1-yl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro- 2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0277] [00277] Prepared from (R) -2-hydroxy-3- (2- (cis-4- (pipera-zin-1-yl) cyclohexyl) acetamido) -3,4-dihydro- 2H-benzo [e] [1,2] oxaborinin-8-carboxylic (Example 17) after the procedure described in Example 4. ESI-MS m / z 458 (MH) +. EXAMPLE 24: (R) -3- (2- (trans-4- (2-guanidinoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1 , 2] oxaborinin-8-carboxylic
[0278] [00278] Synthesis of (R) -3- (2- (trans-4- (2-guanidinoethylami-no) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [ and] [1,2] oxa-borinine-8-carboxylic.
[0279] [00279] Prepared from (R) -3- (2- (trans-4- (2-aminoethyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- benzo [e] [1,2] oxaborinin-8-carboxylic (Example 15) after the procedure described in Example 4. ESI-MS m / z 432 (MH) +. EXAMPLE 25: (R) -2-hydroxy-3- (2- (trans-4- (2-hydroxyethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1.2 ] oxaborinin-8-carboxylic
[0280] [00280] Prepared from (R) -3- (2- (trans-4- (2-aminoethyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- benzo [e] [1.2] oxa-borinine-8-carboxylic (Example 15) and 2- (tert-butyldimethylsilyloxy) acetal-dehyde after the procedure described in Step 1 of Example 15. ESI-MS m / z 505 (MH) +. Step 2: Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (2-hydroxy-tilamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [ and] [1,2] oxaborin-8-carboxylic.
[0281] [00281] (R) -3- (2- (trans-4- (2- (tert-butyldimethylsilyloxy) ethyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro- 2H-benzo [e] [1,2] oxaborinin-8-carboxylic from step 1 (100 mg) in a flask was added 4 ml of 4N HCl in dioxane. The resulting reaction mixture was stirred at RT for 2 h. The solvent was removed in vacuo and the residue was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 391 (MH) +. EXAMPLE 26: (R) -2-hydroxy-3- (2- (trans-4- (pyridin-3-ylmethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1.21oxaborinin-8- (R) -3- (2- (trans-4- (2-carboxylic
[0282] [00282] Synthesis of (R) -3- (2- (trans-4- (2-guanidinoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2Hbenzo acid [e] [1 , 2] oxaborinin-8-carboxylic.
[0283] [00283] To 40 mg of (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [ 1.2] oxaborinin-8-carboxylic (Example 6) in MeOH (5 ml) was added TEA (0.03 ml), followed by nicotinaldehyde (20 mg), AcOH (0.01 ml) and sodium triacetoxyborohydride (25 mg). The reaction mixture was stirred at RT overnight. The solvent was then removed in vacuo and the residue purified by reverse phase HPLC to provide the title compound. ESI-MS m / z 438 (MH) +. EXAMPLE 27: (R) -3- (2- (trans-4- (carboxymethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2 ] oxaborinin-8-carbo-xyl
[0284] [00284] To (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin -8-carboxylic (Example 6) in MeOH was added TEA (2.5 eq), followed by ethyl bromoacetate (1.2 eq). The reaction mixture was stirred at RT overnight. To this reaction mixture was then added 1N NaOH and stirred for 6 h. After concentration in vacuo, 1N HCl was added to adjust the pH to 1. The reaction mixture was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 405 (MH) +. EXAMPLE 28: (R) -3- (2- (trans-4- (4,5-dihydro-1H-imidazol-1-yl) cyclohexyl) acetamido) -2-hydroxy-3,4- acid dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0285] [00285] To (R) -3- (2- (trans-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [ 1.2] oxaborinin-8-carbo-xylate (Example 15) in MeOH was added DIEA (2.5 eq), followed by isopropyl formimidate hydrochloride (1.2 eq). The reaction mixture was stirred at RT overnight. The mixture was then concentrated in vacuo and the crude product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 400 (MH) +. EXAMPLE 29: (R) -3- (2- (trans-4-formimidamidocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin- 8-carboxylic.
[0286] [00286] To (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin -8-carboxylic (Example 6) in MeOH was added DIEA (2.5 eq), followed by isopropyl formimidate hydrochloride (1.2 eq). The reaction mixture was stirred at RT overnight. The mixture was then concentrated in vacuo and the crude product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 374 (MH) +. EXAMPLE 30: (R) -3- (2-cyclohexylacetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic acid
[0287] [00287] Prepared from the 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,0 2,6 ] dec-4-ylmethyl) -benzoic acid and 2-cyclohexylacetic acid after the procedure described in step 1 of Example 1. The crude product was purified by flash chromatography on silica gel (Hexane / EtOAc). Step 2: Synthesis of (R) -3- (3- (trans-4-aminocyclohexyl) propanamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carboxylic.
[0288] [00288] Prepared from 3 - ((2R) -2- (2-cyclohexylacetamido) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6.1 , 1.02.6] dec-4-yl) ethyl) -2-methoxy-benzoate of tert-butyl and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 332 (MH) +. EXAMPLE 31: (R) -2-hydroxy-3- (2- (trans-4- (pyridin-2-ylmethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0289] [00289] Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (pyridin-2-yl-ylamino) cyclohexyl) acetamido) -3,4-dihydro-2H- benzo [e] [1,2] oxaborin-8-carboxylic
[0290] [00290] Prepared from (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] acid [1, 2] oxaborinin-8-carboxylic (Example 6) and picolinaldehyde after the procedure of Example 27. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 438 (MH) +. EXAMPLE 32: (R) -2-hydroxy-3- (2- (trans-4- (piperidin-4-ylmethyl-mino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [ and] [1.21 oxaborinin-8-carboxylic.
[0291] [00291] Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (piperidin-4-ylmethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [ and] [1,2] oxaborinin-8-carboxylic.
[0292] [00292] Prepared from (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1, 2] oxaborinin-8-carboxylic (Example 6) and tert-butyl 4-formylpiperidine-1-carboxylate after the procedure of Example 27. The Boc group was removed by treatment with 4N HCl in dioxane. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 444 (MH) +. EXAMPLE 33: (R) -3- (2- (trans-4 - (((1-carbamimidoylpiperidin-4-yl) methylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H acid -benzo [e] [1,2] oxaborinin-8-carboxylic
[0293] [00293] Synthesis of (R) -3- (2- (trans-4 - (((1-carbamimidoylpiperidin-4-yl) methylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-di acid -hydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0294] [00294] Prepared from (R) -2-hydroxy-3- (2- (trans-4- (piperidin-4-ylmethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H- benzo [e] [1,2] oxaborinin-8-carboxylic (Example 33) and tert-butyl (1H-pyrazol-1-yl) meta-nodiylidenodicarbamate following the procedure of Example 4. The product was purified using phase HPLC reverse to provide the title compound. ESI-MS m / z 486 (MH) +. EXAMPLE 34: (3R) -3- (2- (4- (3-aminoazetidin-1-yl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0295] [00295] Synthesis of (3R) -3- (2- (4- (3-aminoazetidin-1-yl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [ and] [1,2] oxaborinin-8-carboxylic.
[0296] [00296] Prepared from tert-butyl azetidin-3-ylcarbamate after the procedure described in Example 16. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 402 (MH) +. EXAMPLE 35: (3R) -3- (2- (4- (azetidin-3-ylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0297] [00297] Synthesis of (3R) -3- (2- (4- (azetidin-3-ylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carbo-xyl.
[0298] [00298] Prepared from tert-butyl 3-aminoazetidine-1-carboxylate after the procedure described in Example 16. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 402 (MH) +. EXAMPLE 40: (R) -2-hydroxy-3- (2- (4-morpholinocyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8- carboxylic
[0299] [00299] Synthesis of (R) -2-hydroxy-3- (2- (4-morpholinocyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1,2] oxaborinin- 8-carboxylic.
[0300] [00300] Prepared from morpholine and 2- (4-oxocyclohexyl) acetic acid after the procedure described in Example 16. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 417 (MH) +. EXAMPLE 41: (3R) -3- (2- (4- (3-guanidinoazetidin-1-yl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carbo-xylic
[0301] [00301] Synthesis of (3R) -3- (2- (4- (3-guanidinoazetidin-1-yl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [ and] [1,2] oxaborinin-8-carboxylic.
[0302] [00302] Prepared from (3R) -3- (2- (4- (3-aminoazetidin-1-yl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- benzo [e] [1,2] oxaborin-8-carboxylic (Example 35) and tert-butyl (1H-pyrazol-1-yl) methanediylidene-dicarbamate after the procedure of Example 4. The product was purified using Reverse phase HPLC to provide the title compound. ESI-MS m / z 444 (MH) +. EXAMPLE 42: (R) -3 - ((R) -2-amino-2-cyclohexylacetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin acid -8-carboxylic
[0303] [00303] Synthesis of (R) -3 - ((R) -2-amino-2-cyclohexylacetami-do) -2-hydroxy-3,4-dihydro-2H-benzo [e] acid [1] , 2] oxaborinin-8-carboxylic.
[0304] [00304] Prepared from (R) -2- (tert-butoxycarbonyl-mino) -2-cyclohexylacetic acid using the procedure described in step 1 and step 2, Example 1. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 347 (MH) +. EXAMPLE 43: 3- {2- [4- (2-Amino-ethylamino) -1-hydroxy-cyclohexyl] -acetylamino} -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0305] [00305] To a suspension of zinc powder (2.06 g, 31.5 mmol) in diethyl ether (50 mL) under argon was added trimethylsilyl chloride (3.0 mL, 23.6 mmol) and the reaction stirred for 15 min at TA. The reaction was then heated to reflux for 25 min. Benzyl bromoacetate (3.9 mL, 24.6 mmol) and 1,4-cyclohexanedione monoethylene acetal (3.05 g, 19.6 mmol) were added and the reaction was refluxed for 1.3 h. The reaction was then cooled to RT, cooled quickly with 1N HCl (125 ml) and stirred overnight. The aqueous layer was extracted with Et2O (3x). The combined organic layers were washed with sat. NaHCO3, dried over MgSO4, filtered, and concentrated. The crude product was purified by flash silica gel chromatography (0 - 60% EtOAc: Hexane) to provide 2.79 g (54%) of pure product. ESI-MS m / z 285 (M + Na) +.
[0306] [00306] Step 2: Synthesis of benzyl ester of [4- (2-tert-butoxycarbonylamino-ethylamino) -1-hydroxy-cyclohexyl] -acetic acid.
[0307] [00307] Titanium ethoxide (0.47 mL, 2.24 mmol) was added to a solution of benzyl ester of (1-hydroxy-4-oxo-cyclohexyl) -acetic acid (1.22 g, 4, 65 mmol) and 2- (Boc-amino) ethylamine (0.97 g, 6.05 mmol) in DCM (5.0 mL) under argon. The reaction was stirred at RT for 5 h. The reaction was concentrated in vacuo. The residue was diluted with methanol (23 mL) under argon and cooled to -78 ° C. Sodium triacetoxyborohydride (1.49 g, 7.03 mmol) was added in a single portion and the reaction allowed to warm slowly to RT overnight. The reaction was quenched with 10% Na2CO3 and extracted with ethyl acetate (2x). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated. The crude product was carried on without purification. ESI-MS m / z 407 (MH) +. Step 3: Synthesis of {4- [tert-butoxycarbonyl- (2-tert-butoxycarbonylamino-ethyl) -amino] -1-hydroxy-cyclohexyl} -acetic acid benzyl ester.
[0308] [00308] Triethylamine (1.1 mL, 7.89 mmol) and di-tert-butyl dicarbonate (1.22 g, 5.59 mmol) were added to a solution of [4- (2- tert-butoxycarbonylamino-ethylamino) -1-hydroxy-cyclohexyl] -acetic (1.89 g, 4.65 mmol) in DCM (46 mL) under argon. The reaction was stirred at RT for 17 h. The reaction was quenched with brine and extracted with DCM (2x). The combined organic layers were dried over Na2SO4, filtered and concentrated. The crude product was purified by flash silica gel chromatography (0 - 75% EtOAc: Hexane). ESI-MS m / z 507 (MH) +. Step 4: Synthesis of {4- [tert-butoxycarbonyl- (2-tert-butoxycarbon-nylamino-ethyl) -amino] -1-hydroxy-cyclohexyl} -acetic acid.
[0309] [00309] A benzyl ester solution of {4- [tert-butoxycarbonyl- (2-tert-butoxycarbonylamino-ethyl) -amino] -1-hydroxy-cyclohexyl} -acetic acid (0.540 g, 1 , 07 mmol) in methanol (15 mL) was purged with argon for 5 min. Carbon palladium (10%, 0.127 g) was added, the flask evacuated, and the reaction stirred under an H2 atmosphere for 19 h. The reaction was filtered through a plugged Celite filter frit, washed with methanol and DCM, and concentrated. The crude product was carried on without purification. ESI-MS m / z 439 (M + Na) +. Step 5: Synthesis of 3- [2- (2- {4- [tert-butoxy-carbonyl- (2-tert-butoxycarbonylamino-ethyl) -amino] -1-hydroxy-cyclohexyl} tert-butyl ester} -acetylamino) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy -benzoic.
[0310] [00310] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,02,6] dec-4-ylmethyl) -benzoic acid and {4- [tert-butoxycarbonyl- (2-tert-butoxycarbonylamino-ethyl) -amino] -1-hydroxy-cyclohexyl} -acetic acid after the procedure described in step 1 of Example 1. The crude product was purified by flash silica gel chromatography (5-100% EtOAc: Hexane). ESI-MS m / z 828 (MH) +. Step 6: Synthesis of 3- {2- [4- (2-amino-ethylamino) -1-hydroxy-cyclohexyl] -acetylamino} -2-hydroxy-3,4-dihydro-2H-benzo [ and] [1,2] oxaborinin-8-carboxylic.
[0311] [00311] Prepared from 3- [2- (2- {4- [tert-Butoxycarbonyl- (2-tert-butoxycarbonylamino-ethyl) -amino] -1-hydroxy-cyclohexyl} tert-butyl ester} -acetylamino) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy -benzoic and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reversed-phase HPLC and dried using lyophilization. ESI-MS m / z 406 (MH) +. EXAMPLE 44: 3- [2- (4-amino-1-hydroxy-cyclohexyl) -acetylamino] -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin acid -8-carboxylic
[0312] [00312] Prepared from the benzyl ester of (1-hydroxy-4-oxo-cyclohexyl) -acetic acid and benzylamine after the procedure described in Step 2 of Example 43. The crude product was purified by flash silica chromatography. gel (0 - 10% CH3OH: CH2Cl2). ESI-MS m / z 354 (MH) +. Step 2: Synthesis of (4-tert-butoxycarbonylamino-1-hydroxy-cyclohexyl) -acetic acid.
[0313] [00313] A solution of benzyl ester of (4-benzylamino-1-hydroxy-cyclohexyl) -acetic acid (1.41 g, 3.99 mmol) and di-tert-butyl dicarbonate (0.952 g, 4, 36 mmol) in ethanol (35 mL) was purged with argon for 5 min. Palladium hydroxide (20%, 0.608 g) was added, the flask evacuated, and the reaction stirred under an atmosphere of H2 at 65 ° C for 43 h. The reaction was cooled to RT, filtered through a plugged Celite filter frit, washed with methanol and DCM, and concentrated. The crude product was carried on without purification. ESI-MS m / z 296 (M + Na) +. Step 3: Synthesis of 3- [2- [2- (4-tert-butoxycarbonylamino-1-hydroxy-cyclohexyl) -acetylamino] -2- (2,9,9-trimethyl) tert-butyl ester -3,5-dioxa-4-bora-tricyclo [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy-benzoic.
[0314] [00314] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,02,6] dec-4-ylmethyl) -benzoic acid and (4-tert-butoxycarbonylamino-1-hydroxy-cyclohexyl) -acetic acid after the procedure described in step 1 of Example 1. The crude product was purified by flash chromatography on silica gel (5 - 100% EtOAc: Hexane). ESI-MS m / z 685 (MH) +. Step 4: Synthesis of 3- [2- (4-amino-1-hydroxy-cyclohexyl) -acetyl-mino] -2-hydroxy-3,4-dihydro-2H-benzo [e] [1 , 2] oxaborinin-8-carboxylic.
[0315] [00315] Prepared from 3- [2- [2- (4-tert-butoxycarbonylamino-1-hydroxy-cyclohexyl) -acetylamino] -2- (2,9,9-trimethyl) tert-butyl ester -3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy-benzoic and BCl3 after the procedure described in Step 2 of Example 1 The crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 363 (MH) +. EXAMPLE 45: 3- [2- (4-amino-cyclohexylamino) -acetylamino] -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic acid
[0316] [00316] To a suspension of trans-N-Boc-1,4-diaminocyclohexane (0.256 g, 1.19 mmol) and potassium carbonate (0.663 g, 4.80 mmol) in acetonitrile (15 mL) and DMF (5 ml) benzyl bromoacetate (0.21 ml, 1.33 mmol) was added under argon and the reaction was stirred at RT for 19 h. The reaction was diluted with ethyl acetate and washed with sat. and brine. The organic layer was dried over Na2SO4, filtered and concentrated. The crude product was carried on without purification. ESI-MS m / z 363 (MH) +. Step 2: Synthesis of [tert-butoxycarbonyl- (4-tert-butoxycarbonylamino-cyclohexyl) -amino] -acetic acid benzyl ester.
[0317] [00317] Prepared from the benzyl ester of (4-tert-buto-xycarbonylamino-cyclohexylamino) -acetic acid and di-tert-butyl dicarbonate after the procedure described in Step 3 of Example 43. The crude product was purified by silica gel scintillation chromatography (0 - 50% EtOAc: Hexane). ESI-MS m / z 463 (MH) +. Step 3: Synthesis of [tert-butoxycarbonyl- (4-tert-butoxycarbonylamino-cyclohexyl) -amino] -acetic acid.
[0318] [00318] A solution of [tert-butoxycarbon-nil- (4-tert-butoxycarbonylamino-cyclohexyl) -amino] -acetic acid (0.277 g, 0.599 mmol) in methanol (6 mL) was purged with argon for 5 min. Palladium hydroxide (20%, 0.053 g) was added, the flask evacuated, and the reaction stirred under an H2 atmosphere for 19 h. The reaction was filtered through a plugged Celite filter frit, washed with methanol and DCM, and concentrated. The crude product was carried on without purification. ESI-MS m / z 395 (M + Na) +. Step 4: Synthesis of 3- [2- {2- [tert-butoxycarbonyl- (4-tert-butoxycarbonylamino-cyclohexyl) -amino] -acetylamino} -2- (2.9) , 9-trimethyl-3,5-dioxa-4-bora-tricyclo [6.1, 1.02.6] dec-4-yl) -ethyl] -2-methoxy-benzoic.
[0319] [00319] Prepared from the tert-butyl ester of 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,026] dec-4- ilmethyl) -benzoic acid and [tert-butoxycarbonyl- (4-tert-butoxycarbonylamino-cyclohexyl) -amino] -acetic acid after the procedure described in step 1 of Example 1. The crude product was purified by flash chromatography on silica gel (5 - 100% EtOAc: Hexane). ESI-MS m / z 784 (MH) +. Step 5: Synthesis of 3- [2- (4-amino-cyclohexylamino) -acetylamino] -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8 acid -carboxylic.
[0320] [00320] Prepared from 3- [2- {2- [tert-butoxycarbonyl- (4-tert-butoxycarbonylamino-cyclohexyl) -amino] -acetylamino} -2- (tert-butyl ester) -2- (2.9 , 9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) -ethyl] -2-methoxy-benzoic and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reversed-phase HPLC and dried using lyophilization. ESI-MS m / z 362 (MH) +. EXAMPLE 46: (R) -3- (2- (cis-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1 , 2] oxaborinin-8-carboxylic
[0321] [00321] Synthesis of (R) -3- (2- (cis-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-acid.
[0322] [00322] Prepared from (R) -3- (2- (cis-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1, 2] oxaborinin-8-carbo-xyl (Example 7) after the same procedure as described in Example 10. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 390 (MH) +. EXAMPLE 47: (3R) -2-hydroxy-3- (2- (4-hydroxycyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8- carboxylic
[0323] [00323] Prepared from the 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,0 26] dec -4-ylmethyl) -benzoic acid and 2- (4-oxocyclohexyl) acetic acid after the procedure described in step 1 of Example 1. The crude product was purified by flash chromatography on silica gel (Hexane / EtOAc). ESI-MS m / z 568.1 (MH) +. Step 2: Synthesis of (3R) -2-hydroxy-3- (2- (4-hydroxycyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1,2] oxaborinin- 8-carboxylic.
[0324] [00324] (R) -2-Hydroxy-3- (2- (4-oxocyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8- carboxylic acid was prepared from 3 - ((2R) -2- (hexahydrobenzo [d] [1,3,2] dioxaborol-2-yl) -2- (2,9,9-trimethyl-3,5-dioxa-4 -bora-tricycle [6,1,1,026] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate as described in Step 2, Example 1. To the crude product in H2O and MeOH was added NaBH4. The resulting reaction mixture was stirred at room temperature for 4 h. After removing MeOH, the product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 348 (MH) +. EXAMPLE 48: (R) -2-hydroxy-3- (2- (trans-4- (2- (pyridin-2-ylamino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H -benzo [e] [1,2] oxaborinin-8-carboxylic.
[0325] [00325] To a mixture of 3-carboxy-2-methoxy-phenyl boronic acid (35.0 g, 178.5 mmol,) and (+) pinanediol (30.35 g, 178.5 mmol) was added toluene ( 400 mL). The resulting mixture was stirred for 3 h then concentrated in vacuo (28 mmHg, bath temperature 40 ° C). The resulting solid was dried by toluene azeotrope (2 times, approximately 100 ml). This residue was dried under high vacuum (approx. 1 mmHg) at room temperature for 17.5 h to provide the crude title compound which can be used without further purification. 48.06 g of this crude product was recrystallized from 150 ml of chloroform / hexane (1: 5 v / v) to provide the crude title compound. The mother liquor for crystallization was concentrated and purified by silica chromatography (120 g of silica eluted with 40-100% ethyl acetate in hexane) to provide an additional batch of the title compound. Step 1b: Synthesis of 3-tert-butyloxycarbonyl-2-methoxy-phenyl boronic acid (+) pinanediolate.
[0326] [00326] Recrystallized 3-carboxy-2-methoxy-phenyl boronic acid (+) pinanediolate diester (9.90 g, 30 mmol) was added thionyl chloride (20 mL, reagent grade) and the reaction flask ventilated through a CaCl2 siphon. The resulting solution was heated in an oil bath maintained at 95 ° C and stirred for 1 h. This solution was cooled to room temperature for about 10 min, then concentrated under reduced pressure (20 - 30 mm H g, 35 ° C) until a constant weight of 10.57 g was reached.
[0327] [00327] In a separate vial, to a cooled (-5 ° C) solution of t-BuOH (3.7 mL, 39 mmol) in THF (100 mL) was added, by dripping, BuLi (14.4 mL, 2.5 M in hexane, 36 mmol) for about 5 min. On complete addition, the resulting solution was stirred for 20 min. To this solution was added the crude acid chloride (above) (10.57 g, 30 mmol) in THF (15 ml) for about 30 sec. Upon complete addition, the cold bath was removed and stirring was continued for 4 h. To this solution was added HCl (50 ml, 0.2 M aq). The mixture (pH = 3) was extracted with ether and the ether extract washed with brine, dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (120 g silica eluted with 2 -20% ethyl acetate in hexane) to provide the title compound as a cream colored solid. Step 1c: Synthesis of methylboronic acid (+) pinanediolate (3-tert-butoxycarbonyl-2-methoxy-phenyl) diester.
[0328] [00328] To a cooled solution (-100 ° C outside temperature) of 3-ether-butyloxycarbonyl-2-methoxy-phenyl boronic acid (+) pinanediolate diester (34.6 g, 89.6 mmol) and chlorine- iodomethane (10.3 ml, 140 mmol) in THF (250 ml) was added, dropwise from the side of the flask, BuLi (54 ml, 2.5 M in hexanes, 135 mmol) over 80 min. On complete addition, the reaction solution was stirred 15 min. To the resulting solution, ZnCl2 (90 mL, 1M in ether) was added by dripping from the side of the flask for approximately 40 min. Upon complete addition, the cold bath was removed and stirring continued for 16.5 h. The reaction mixture was diluted with NH4Cl (300 ml, aqueous saturated) and ethyl acetate (700 ml). The separated organic extract was washed with another portion of saturated aq NH4Cl (100 ml) and brine (100 ml), dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (224 g of silica eluted with hexane (1L), then 10% ethyl acetate in hexane (2 L) to provide the title compound as a colorless oil. This material was slowly crystallized at - 10 ° C. Step 1d: Synthesis of boronic [(1S) -2- (3-ether-butoxycarbonyl-2-methoxy-phenyl) -1-chloro-ethyl] boronic diester
[0329] [00329] Method 1: To a cooled (-100 ° C outside temperature) solution of dichloromethane (2.27 mL, 35 mmol) in THF (44 mL) was added, by dripping from the side of the flask, BuLi (8.88 mL, 2.5 M in hexanes, 22 mmol) for 45 min. After approx. 80% of the BuLi was added, a white precipitate formed. Upon complete addition, the reaction mixture was stirred 30 min. To this mixture was added, by dripping from the side of the flask, (+) pinanediolate diester (3-tert-butoxycarbonyl-2-methoxy-phenyl) methylbohonic acid (8.0 g, 20 mmol) in THF (20 mL) for approximately 30 min. Upon complete addition, the resulting solution was stirred for 5 minutes. To this solution was added ZnCh (22 mL, 1M in ether) by dripping from the side of the flask for approximately 12 min. Upon complete addition, the cold bath was removed and replaced with a bath at -10 ° C. The reaction mixture was stirred for 1.25 h. To this solution was added chilled ether (300 ml) and chilled aqueous saturated NH4Cl (125 ml). The organic phase was washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by chromatography on silica (120 g silica eluted with 2 -20% ethyl acetate in hexane) to provide the title compound as a colorless oil. This material was slowly crystallized at -10 ° C.
[0330] [00330] Method 2: methylboronic acid (3-tert-butoxycarbonyl-2-methoxy-phenyl) (+) pinanediolate diester (2.0 g, 5 mmol) and dichloromethane (1.6 mL, 25 mmol) in THF (20 ml) was stirred at -60 ° C for 30 min. To this solution was added LDA (6.5 mmol, 2 M Aldrich's solution) over a period of 10 min. The resulting reaction mixture was stirred at -60 ° C for 20 min. ZnCl2 (8.75 mmol, 1M ether solution) was added at -60 ° C slowly. The reaction mixture was stirred at -50 to -60 ° C for 30 min. This resulting mixture was heated to 0 ° C over a period of 1 h, at which time 10% H2SO4 solution (10 mL) was added and the reaction mixture stirred for 10 min. After phase separation, the organic phase was washed with water and brine. The organic phase was then dried and concentrated in vacuo. The residue was then purified by flash silica chromatography (EtOAc / Hexane: 4/1) to provide the title compound. Step 2: Synthesis of ethyl 2- [4- [2- (2-pyridylamino) ethylamino] cyclohexyl] acetate.
[0331] [00331] To a mixture of 2- (N- [2- (amino) -ethyl] -amino) -pyridine (685 mg, 5 mmol) and 2- (4-oxocyclohexyl) ethyl acetate (786 mg, 4 mmol) dichloromethane (4 mL) was added followed by titanium ethoxide (420 μΙ, 2 mmol, technical grade). The resulting mixture was stirred for 4 h, then concentrated under reduced pressure. The residue was taken up in methanol (10 ml) and cooled to -78 ° C. To this solution was added sodium borohydride (228 mg, 6 mmol). Upon complete addition, the cold bath was removed and stirring continued for 1.25 h. The mixture was diluted with dichloromethane and poured into saturated sodium carbonate solution (15 ml). The organic phase was separated, dried over magnesium sulfate and concentrated to provide the crude title compound as a 6: 1 mixture of trans: cis isomers. This mixture was used without further purification. Step 3: Synthesis of ethyl 2- [4- [tert-butoxycarbonyl- [2- [tert-butoxycarbonyl (2-pyridyl) amino] ethyl] amino] cyclohexyl] acetate.
[0332] [00332] To a solution of 2- [4- [2- (2-pyridylamino) ethylamino] cyclohexyl] ethyl acetate (1.31 g, 4 mmol) in dichloromethane (12 mL) was added di-ether- butyl-dicarbonate (2.18 g, 10 mmol) followed by diisopropylethylamine (1.76 mL, 10 mmol). The resulting solution was stirred for 4 h, diluted with ethyl acetate, washed with brine, dried over magnesium sulfate and concentrated. The residue was purified by silica chromatography (30 g silica; eluted with 20% ethyl acetate / 10% dichloromethane in hexanes) to provide the title compound as a 6: 1 mixture of trans: cis isomers. Step 4: Synthesis of 2- [4- [tert-butoxycarbonyl- [2- [tert-butoxycarbonyl (2-pyridyl) amino] ethyl] amino] cyclohexyl] acetic acid.
[0333] [00333] To a solution of 2- [4- [tert-butoxycarbonyl- [2- [tert-butoxy-carbonyl (2-pyridyl) amino] ethyl] amino] cyclohexyl] ethyl acetate (968 mg, 1, 91 mmol) in THF (3 mL); methanol (3 ml) and water (6 ml) lithium hydroxide monohydrate (397 mg, 9.7 mmol) was added. The resulting solution was stirred for 2.25 h, then acidified to pH 3 with 1 N HCl. The resulting mixture was extracted with dichloromethane (4 times). The combined organic extract was dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (10 g silica; eluted with 40 - 100% ethyl acetate in hexanes) to provide the title compound as a 6: 1 mixture of trans: cis isomers. Step 5: Synthesis of 2-methoxy-3 - ((2R) -2- (2- (trans-4- (2- (pyridin-2-ylamino) ethylamino) cyclohexyl) acetamido) -2- (2, 9,9-trimethyl-3,5-dioxa-4-boron-tri-cyclo [6,1,1,02,6] dec-4-yl) ethyl) tert-butyl benzoate.
[0334] [00334] To a cooled (-78 ° C) solution of [(1S) -2- (3-ferc-butoxycarbonyl-2-methoxy-phenyl) -1-chloro-ethyl] boronic acid (+) pinanediolate (from step 1) (1.35 g, 3 mmol) in THF (9 mL) a solution of lithium bistrimethylsilyl amide (3.0 mL, 1M in THF, 3 mmol) was added dropwise. Upon complete addition, the cold bath was removed and stirring continued for 16.75 hours. The resulting solution, which was approximately 0.25 M benzoic acid, 3 - [(2R) -2- [bis (trimethylsilyl) amino] -2 - [(3aS, 4S, 6S, 7aR) -hexahydro-3a, 5, 5-trimethyl-4,6-methane-1,3,2-benzodioxaborol-2-yl] ethyl] -2-methoxy, 1,1-dimethyl ethyl ester in THF, was used without further purification.
[0335] [00335] In a separate bottle, in a mixture of 2- [4- [tert-butoxycarbonyl- [2- [tert-butoxycarbonyl (2-pyridyl) amino] ethyl] amino] cyclohexyl] acetic acid (477 mg, 1 mmol) and HATU (418 mg, 1.1 mmol) was added DMA (3mL) followed by N-methyl-morpholine (120 μl, 1.1 mmol). The resulting solution was stirred for 90 minutes. To this solution was added a solution of benzoic acid, 3 - [(2R) -2- [bis (trimethylsilyl) amino] -2 - [(3aS, 4S, 6S, 7aR) - hexahydro-3a, 5,5-trimethyl -4,6-methane-1,3,2-benzodioxaborol-2-yl] ethyl] -2-methoxy, 1,1-dimethylethyl ester (4 mL, 0.25 M in 1 mmol THF). The resulting mixture was stirred for 2.5 hours, diluted with ethyl acetate, washed with water and brine, dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (10 g silica; eluted with 20 - 100% ethyl acetate in hexanes) to provide the title compound. Step 6: Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (2- (pyridin-2-ylamino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro -2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0336] [00336] To a solution of 2-methoxy-3 - ((2R) -2- (2- (trans-4- (2- (pyridin-2-ylamino) ethylamino) cyclohexyl) acetamido) -2- ( 2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) tert-butyl benzoate (396 mg, 0.45 mmol) in 1,4-dioxane (6 ml) HCl (6 ml, 3M in water) was added. The resulting solution was heated to reflux and stirred at this temperature for 3.5 hours. The resulting mixture was cooled to room temperature and extracted with ether (2X). The remaining aqueous solution was purified directly by Phenomenex Luna C18 35 x 75 mm reverse phase HPLC; flow rate 40 mL / min; eluted with 5 - 70% CH3CN in H2O / 0.1% TFA for 8 minutes. The title compound was isolated as the TFA salt by lyophilization. EXAMPLE 49: (R) -2-hydroxy-3- (2- (trans-4- (2- (methylsulfonamido) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e ] [1,2] oxaborinin-8-carboxylic
[0337] [00337] To a cooled (-10 ° C) solution of N- (2-aminoethyl) 2-tert-butyl carbeate (1.60 g, 10 mmol) in dichloromethane (25 mL) was added triethylamine (1 , 38 mL, 10 mmol) followed by methanesulfonyl chloride (770 μl, 10 mmol). The resulting solution was stirred for 5 minutes, then the cold bath was removed and stirring continued for 1 h. The reaction mixture was then diluted with ethyl acetate, washed with water and brine, dried over magnesium sulfate and concentrated to provide the title compound as a solid. This material was used without further purification. Step 2: Synthesis of N- (2-aminoethyl) methanesulfonamide; 2.2.2-trifluoroacetic acid salt.
[0338] [00338] To a solution of tert-butyl N- [2- (methanesulfonamido) ethyl] carbamate (2.23 g, 9.4 mmol) in dichloromethane (30 mL) was added trifluoroacetic acid (7.5 mL). The resulting solution was stirred for 1.5 h, then concentrated in vacuo to provide the title compound as a white solid. This material was used without further purification. Step 3: Synthesis of ethyl 2- [4- [2- (methanesulfonamido) ethylamino] cyclohexyl] acetate.
[0339] [00339] To a mixture of ethyl 2- (4-oxocyclohexyl) acetate (736mg, 4 mmol) and N- (2-aminoethyl) methanesulfonamide; 2.2.2-trifluoroacetic acid salt (1.26 g, 5 mmol) in dichloromethane (6 mL) was added triethylamine (690 μl 5 mmol) followed by titanium (IV) ethoxide (420 μl 2 mmol, technical grade). The resulting cloudy mixture was stirred for 4 hours, then concentrated under reduced pressure. The residue was taken up in methanol (6 ml). This mixture was cooled to -78 ° C. To the resulting mixture, sodium borohydride (187mg, 4.8 mmol) was added. Upon complete addition, the cold bath was allowed to expire and stirring continued for 15.5 hours. The resulting mixture was concentrated under reduced pressure to provide a thick mass. This residue was taken up in dichloromethane (40 ml). To this mixture was added Na2CO3 (5.5 ml, saturated aqueous solution). The resulting mixture was stirred for 5 minutes. To this mixture was added celite (1.8 g). This mixture was stirred for 5 minutes, then filtered through a pad of celite. The filtrate is washed with saturated aqueous Na2CO3, dried over magnesium sulfate and concentrated to provide the title compound as a 6: 1 mixture of trans: cis isomers. This material was used without further purification. Step 4: Synthesis of 2- [4- [tert-butoxycarbonyl- [2- (methanesulfonamido) ethyl] amino] cyclohexyl] ethyl acetate
[0340] [00340] To a solution of 2- [4- [2- (methanesulfonamido) ethylamino] cyclohexyl] ethyl acetate (1.1 g, 4 mmol) in dichloromethane (12 mL) was added di-tert- butyl-dicarbonate (1.74 g, 8 mmol) followed by diisopropylethylamine (1.4 mL, 8 mmol). The resulting solution was stirred for 4 hours, then diluted with dichloromethane, washed with water, dried over magnesium sulfate and concentrated. The residue was purified by silica chromatography (25 g silica; eluted with 5 - 50% ethyl acetate in hexanes) to provide the title compound as a 6: 1 trans: cis isomer. Step 5: Synthesis of trans 2- [4- [tert-butoxycarbonyl- [2- (meta-nosulfonamido) ethyl] amino] cyclohexyl] acetic acid.
[0341] [00341] To a solution of 2- [4- [tert-butoxycarbonyl- [2- (methanesulphonamido) ethyl] amino] cyclohexyl] ethyl acetate (0.842 g, 2.06 mmol) in methanol (4 mL ); THF (4 ml) and water (4 ml) were added lithium hydroxide monohydrate (252 mg, 6 mmol). The resulting solution was stirred for 2 hours, then acidified with HCl (7 ml, 1M aq.), Diluted with ethyl acetate, washed with brine, dried over magnesium sulfate and concentrated. The residue was triturated with ether to provide the title compound as a solid. Step 6: Synthesis of acid (+) pinanediolate of [(1R) -1 - [[2- [4- [ferc-butoxycarbonyl- [2- (methanesulfonamido) ethyl] amino] cyclohexyl] acetyl] amino] -2- (3-fer-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic
[0342] [00342] The title compound was prepared using the same procedure as described in Example 48; Step 4 unless using 2- [4- [tert-butoxycarbonyl- [2- (methanesulfone-mido) ethyl] amino] cyclohexyl] acetic acid instead of 2- [4- [tert-butoxycarbonyl) - [2- [tert-butoxycarbonyl (2-pyridyl) amino] ethyl] amino] cyclohexyl] acetic. Step 7: Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (2- (methylsulfo-namido) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H -benzo [e] [1,2] oxaborinin-8-carboxylic.
[0343] [00343] To a solution of [(1R) -1 - [[2- [4- [tert-butoxycarbonyl- [2- (methanesulfonamido) ethyl] amino] cyclohexyl] acetyl] (+) pinanediolate diester amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic (460 mg, 0.55 mmol) in 1,4-dioxane (4 mL) HCl (4 mL, 3 M in water). The resulting solution was heated to reflux and stirred at this temperature for 3.5 hours. The resulting mixture was cooled to room temperature and extracted with ether (2 times). The remaining aqueous solution was concentrated to 25% by volume and the residue was directly purified by reverse phase HPLC Column Phenomenex Luna C18 35 x 75 mm; flow rate 40 mL / min; eluted with 5 - 70% CH3CN in H2O / 0.1% TFA for 8 minutes. The title compound was isolated as the TFA salt by lyophilization. EXAMPLE 50: (S) -3- (2- (trans-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1 , 2] oxaborinin-8-carbo-xylic.
[0344] [00344] To a solution of 3-iodo-2-hydroxy-benzaldehyde (4.0 g, 16.06 mmol) in DMA (32 mL) was added cesium carbonate (5.85 g, 18 mmol) followed by iodide methyl (1.12 mL, 18 mmol). The resulting mixture was stirred for 4.75 hours, then diluted with ether, washed with water, dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (50 g silica; eluted with 0 - 20% ethyl acetate in hexanes) to provide the title compound as an oil. Step 2: Synthesis of 3-iodo-2-methoxy-benzoic acid.
[0345] [00345] To a solution of 3-iodo-2-methoxy-benzaldehyde (3.68 g, 14 mmol) in tert-butanol (70 mL) was added 2,3-dimethyl-but-2-ene (7 mL) followed by a solution comprising disodium phosphate hydrogen monohydrate (7.56 g 56 mmol) and sodium chloride (7.56 g, approx. 66 mmol, technical grade) in water (70 mL). The resulting mixture was stirred for 20 minutes, then diluted with ethyl acetate, washed with brine, dried over magnesium sulfate and concentrated. The residual solid was recrystallized from cyclohexane to provide the title compound as a white solid. Step 3: Synthesis of 3-iodo-2-methoxy-benzoyl chloride.
[0346] [00346] To 3-iodo-2-methoxy-benzoic acid (8.29 g, 29.8 mmol) was added thionyl chloride (15 mL). The resulting solution was stirred for 2 minutes then heated to 82 ° C and stirred at this temperature for 30 minutes. The solution was then concentrated under reduced pressure to provide the title compound. This material was used without further purification. Step 4: Synthesis of tert-butyl 3-iodo-2-methoxy-benzoate.
[0347] [00347] To a cooled (-5 ° C) solution of tert-butanol (12.87 mL, 30 mmol) in THF (30 mL) was added, by dripping, BuLi (12.0 mL, 2.5 M in hexanes, 30 mmol). Upon complete addition, the solution was stirred for 20 minutes. To this solution is added a solution of 3-iodo-2-methoxy-benzoyl chloride (8.8 g, 29.8 mmol) in THF (12 mL). Upon complete addition, the cold bath was removed and stirring continued for 1.5 h. This solution was diluted with ethyl acetate, washed with water and brine, dried over magnesium sulfate and concentrated. The residue was purified by silica chromatography (50 g silica; eluted with 0 - 20% ethyl acetate in hexanes) to provide the title compound as an oil. Step 5: Synthesis of methylboronic acid (3-tert-buto-xicarbonyl-2-methoxy-phenyl) (-) pinanediolate diester
[0348] [00348] To a cooled (-40 ° C) solution of tert-butyl 3-iodo-2-methoxy-benzo-act (3.34 g, 10 mmol) in THF (25 mL) was added, by dripping, isopropylmagnesium chloride complex: lithium chloride (7.69 mL, 1.3 M in THF, 10 mmol). Upon complete addition, the solution was stirred for 20 minutes, then cooled to -78 ° C. To this solution was added, by dripping from the side of the flask, chloro-methylboronic acid (-) pinanediolate diester (2.28 g, 10 mmol) in THF (2 mL) (chloro-acid (-) pinanediolate diester) Methylboronic was prepared according to Strynadka et al. Biochemistry 2000, 39, 5312; unless using (-) pinanediol instead of (+) pinanediol). The resulting solution was stirred for 45 minutes. ZnCl2 (10 mL, 1M in ether, 10 mmol) was added to this solution. The resulting mixture was stirred for 5 minutes, then the cold bath was removed and stirring continued for 2 hours. This mixture was diluted with ether, washed with 0.1 M HCl and brine, dried over magnesium sulfate and concentrated. The residue is purified by chromatography on silica (50 g silica; eluted with 0 - 20% ethyl acetate in hexanes) to provide the title compound as an oil. This material crystallizes on standing at -10 ° C. Step 6: Synthesis of [(1R) -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) -1-chloro-ethyl] boronic acid (-) pinanediolate
[0349] [00349] To a cooled (-100 ° C) solution of dichloromethane (518 μΙ, 8 mmol) in THF (10 mL) was added, by dripping from the side of the flask, BuLi (2.0 mL, 2.5 M in hexanes, 5 mmol) for about 20 minutes. A precipitate forms after about 75% of the BuLi has been added. Upon complete addition, the resulting cloudy solution was stirred for 40 minutes. To this mixture, a solution of (-) pinanediolate diester (3-tert-butoxycarbonyl-2-methoxy-phenyl) methylboronic acid (1.8 g, 4.5 mmol) was added by dripping from the side of the flask. THF (4 mL). On complete addition, a solution of ZnCl2 (5 mL, 1 M in ether, 5 mmol) was added, dropwise, for about 8 minutes. The -100 ° C bath was replaced with a -10 ° C bath and the resulting mixture was allowed to stir for 1 hour. To this mixture was added a cold saturated solution of NH4Cl followed by cold ether (5 ° C). The organic phase was separated, washed with brine, dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (25 g silica; eluted with 0 - 20% ethyl acetate in hexanes) to provide the title compound as an oil. This material crystallizes on standing at -10 ° C. Step 7: Synthesis of 2- [4- [tert-butoxycarbonyl- [3- (tert-butoxycarbonyl-mino) ethyl] amino] cyclohexyl] ethyl acetate.
[0350] [00350] The title compound (isolated as a 6.8: 1 mixture of trans: cis isomers) was prepared using essentially the same procedure used in Example 48, Step 2, unless using ethyl 2- [4- [ Ethyl 3- (tert-butoxycarbonylamino) ethylamino] cyclohexyl] acetate instead of 2- [4- [2- (2-pyridylamino) ethylamino] cyclohexyl] ethyl acetate. Step 8: Synthesis of trans-2- [4- [tert-butoxycarbonyl- [3- (tert-buto-xicarbonylamino) ethyl] amino] cyclohexyl] acetic acid.
[0351] [00351] The title compound was prepared using essentially the same procedure used in Example 63, Step 3, unless using 2- [4- [ferc-butoxycarbonylamino] [3- (tert-butoxycarbonylamino) ethyl] amino] cycle -hexyl] ethyl acetate instead of 2- [4- [tert-butoxycarbonyl- [3- (tert-butoxycarbonylamino) propyl] amino] cyclohexyl] ethyl acetate. Step 9: Synthesis of acid (-) pinanediolate of [(1S) -1 - [[2- [trans-4- [tert-butoxycarbonylamino) ethyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic
[0352] [00352] To a cooled (-20 ° C) solution of [(1R) -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) -1-chloro-ethyl] boronic acid (-) pinanediolate (430 mg, 1 mmol) in THF (2 mL), a solution of lithium bistrimethylsilyl amide (1.0 mL, 1M in THF, 1 mmol) was added dropwise. Upon complete addition, the cold bath was removed and stirring was continued for 1 hour. The resulting solution in approximately 0.29 M [(1S) -1- [bis (trimethylsilyl) amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic acid (-) pinanediolate was used without another procedure.
[0353] [00353] In a separate vial, in a mixture of 2- [4-trans- [tert-butoxycarbonyl- [2- (tert-butoxycarbonylamino) ethyl] amino] cyclohexyl] acetic acid (400 mg, 1 mmomL) and HATU (418 mg, 1.1 mmol) was added DMA (2 mL) followed by N-methyl-morpholine (120 μl, 1.1 mmol). The resulting solution was stirred for 90 minutes. To this solution was added the solution of approximately 0.29 M diester of (-) pinanediolate of [(1S) -1- [bis (trimethylsilyl) amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl] ) ethyl] boronic acid prepared above. The resulting mixture was stirred for 4 hours, diluted with ethyl acetate, washed with water and brine, dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (10 g silica; eluted with 20 - 100% ethyl acetate in hexanes) to provide the title compound as a foam. Step 10: Synthesis of (S) -3- (2- (trans-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0354] [00354] To a cooled solution (-78 ° C) of acid (-) pinanediolate diester [(1S) -1 - [[2- [trans-4- [tert-butoxycarbonyl- [2- (tert-butoxycarbonylamino ) ethyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic (468 mg, 0.59 mmol) in dichloromethane (2 mL) was added, by dripping, a solution of BCl3 (3 mL, 1M in CH2Cl2, 3 mmol). Upon complete addition, the resulting mixture was stirred for 30 minutes then heated to 0 ° C for 30 minutes. To this mixture, water (6 ml) was added. The resulting mixture was allowed to warm to room temperature for 20 minutes. This mixture was extracted with ether and the remaining aqueous phase was purified by reverse phase HPLC (Phenomenex Luna C18 35 x 75 mm column; Flow rate 40 mL / min; eluted with 5 - 45% CH3CN in H2O / 0.1% TFA for 8 minutes). The title compound was isolated as the TFA salt by lyophilization. ESI-MS m / z 390 (MH) +. EXAMPLE 51: (R) -2-hydroxy-3- (2- (trans-4- (2- (methylamino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e ] [1,2] oxaborinin-8-carboxylic
[0355] [00355] Prepared from 2- (trans-4- (tert-butoxycarbonyl (2- (tert-butoxycarbonyl (methyl) amino) ethyl) amino) cyclohexyl) acetic acid using the procedure described in step 1 and step 2 , Example 1. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 404 (MH) +. EXAMPLE 52: (R) -2-hydroxy-3- (2- (trans-4- (2-imino-3-methylimide-zolidin-1-yl) cyclohexyl) acetamido) -3,4-di- hydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0356] [00356] Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (2-imino-3-methyl-limidazolidin-1-yl) cyclohexyl) acetamido) -3,4- dihydro-2H-benzo [e] [1.21 oxaborinin-8-carboxylic.
[0357] [00357] To (R) -2-hydroxy-3- (2- (trans-4- (2- (methylamino) ethyl-mino) cyclohexyl) acetamido) -3,4-dihydro-2H- benzo [e] [1.21oxaborinin-8-carboxylic acid from Example 51 (10 mg) in MeOH (1 ml) was added tert-butyl methanediylidenodicarbamate (1H-pyrazol-1-yl) (12 mg) and TEA (0.1 ml) . The reaction mixture was stirred at RT for 48 h. The solvent was removed in vacuo. The residue was treated with a mixture of TFA (3 ml) and DCM (2 ml) and stirred for 1 h. The solvent was then removed in vacuo and the crude product was purified by preparative reversed-phase HPLC and dried using lyophilization. ESI-MS m / z 429 (MH) +. EXAMPLE 53: (R) -3- (2- (trans-4 - ((S) -2-aminopropylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [ e] [1.21 oxaborinin-8-carboxylic
[0358] [00358] Ao 3 - ((2R) -2- (2- (tran-4- (tert-butoxycarbonylamino) cyclohexyl) acetamido) -2- (2.9.9-trimethyl-3,5-dioxa-4- bora-tricycle [6.1.1.02.6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate from step 1 of Example 6 (640 mg) 4 N HCl in dioxane (4 ml) was added. The resulting reaction mixture was stirred at RT for 1 h. Diethyl ether was added to the precipitate outside the product as a white solid (500 mg) which was used directly in the next step. Step 2: Synthesis of acid 3 - ((2R) -2- (2- (trans-4 - ((S) -2- (tert-butoxycarbonylamino) propylamino) cyclohexyl) acetamido) -2- (2 , 9,9-trimethyl-3,5-dio-xa-4-bora-tricyclo [6.1, 1.02.6] dec-4-yl) ethyl) -2-methoxybenzoic.
[0359] [00359] Prepared from acid 3 - ((2R) -2- (2- (trans-4-amino) cyclohexyl) acetamido) -2- (2,9,9-trimethyl-3,5-dioxa -4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-methoxybenzoic and (S) -tert-butyl 1-oxopropan-2-ylcarbamate after the procedure described in Example 27. Step 3: Synthesis of (R) -3- (2- (trans-4 - ((S) -2-aminopropylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- benzo [e] [1,2] oxaborinin-8-carboxylic. To 3 - ((2R) -2- (2- (trans-4 - ((S) -2- (tert-butoxycarbonylamino) propylamino) cyclohexyl) acetamido) -2- (2,9,9-trimethyl) -3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-methoxybenzoic from step 2 in a flask 3 N aqueous HCl was added and the mixture The reaction mixture was stirred at reflux for 1 h. The product was purified using reverse phase HPLC and dried using lyophilization. ESI-MS m / z 409 (MH) +. EXAMPLE 54: (R) -2-hydroxy-3- (2- (trans-4- (2- (methoxycarbonyl-mino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo acid [e] [1,2] oxa-borinine-8-carboxylic
[0360] [00360] Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (2- (methoxycarbonylamino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H -benzo [e] [1,2] oxaborinin-8-carboxylic.
[0361] [00361] To (R) -2-hydroxy-3- (2- (trans-4- (2- (methylamino) ethyl-mino) cyclohexyl) acetamido) -3,4-dihydro-2H- benzo [e] [1,2] oxaborinin-8-carboxylic acid of Example 51 (39 mg) in a mixture of H2O (1 ml), THF (1 ml) and MeOH (1 ml) was added NaHCO3 (200 mg), followed by methyl chloroformate (1.2 eq). The reaction mixture was stirred at RT overnight. The solvent was then removed in vacuo and the crude product was purified by preparative reversed-phase HPLC and dried using lyophilization. ESI-MS m / z 448 (MH) +. EXAMPLE 55: (R) -3- (2- (3- (2-aminoethylamino) cyclobutyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin -8- carboxylic
[0362] [00362] To methyl 2- (3-oxocyclobutyl) acetate (284 mg) and tert-butyl 2-amino-ethylcarbamate (336 mg) in a flask, MeOH (10 mL) and Pd / C (10%) were added , 50 mg). The reaction mixture was stirred under a hydrogen atmosphere overnight. At the end of the reaction, the catalyst was filtered through the Celite buffer and the solvent was removed under reduced pressure. The crude product (577 mg) was taken to the next step without further purification. Step 2: Synthesis of methyl 2- (3- (tert-butoxycarbonyl (2- (tert-butoxycarbonyl-mino) ethyl) amino) cyclobutyl) acetate.
[0363] [00363] To the product from step 1 in DCM (15 ml) was added TEA (0.35 ml) and di-tert-butyl dicarbonate (480 mg). The reaction mixture was stirred at RT overnight. The organic phase was washed with 1 N HCl, water and brine, dried over sodium sulfate. Removal of solvents under reduced pressure gave the product (1.0 g) without further characterization. Step 3: Synthesis of 2- (3- (tert-butoxycarbonyl (2- (tert-butoxycarbonyl-amino) ethyl) amino) cyclobutyl) acetic acid.
[0364] [00364] To 2- (3- (tert-butoxycarbonylamino) (2- (tert-butoxycarbonylamino) ethyl) amino) cyclobutyl) methyl acetate from step 2 in a mixture of MeOH and H2O was added 1N NaOH (8 mL). The resulting reaction mixture was stirred at RT overnight. Half of the solvents were removed under reduced pressure and 1N HCl was added to adjust the pH of the solution to 4. The aqueous phase was extracted with EtOAc three times. The combined organic phase was then dried and concentrated in vacuo to provide the acid (0.9 g). Step 4: Synthesis of 3 - ((R) -2- (2- (3- (tert-butoxy-carbonylamino) ethyl) amino) cyclobutyl) acetamido) -2- (2.9, Tert-butyl 9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-methoxybenzoate
[0365] [00365] Prepared from the 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,0 2,6 ] dec-4-ylmethyl) -benzoic acid and 2- (3- (tert-butoxycarbonyl (2- (tert-butoxycarbonyl-mino) ethyl) amino) cyclobutyl) acetic acid after the procedure described in Step 1 of Example 1 The crude product was purified by silica gel scintillation chromatography (hexane / EtOAc, 2: 1 to 1: 2). ESI-MS m / z 784.1 (MH) +. Step 5: Synthesis of (R) -3- (2- (3- (2-aminoethylamino) cyclobutyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2 ] oxaborinin-8-carboxylic.
[0366] [00366] Prepared from 3 - ((R) -2- (2- (3- (tert-butoxycarbonylamino) ethyl) amino) cyclobutyl) acetamido) -2- (2.9, 9-tri-methyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate and BCl3 after the described procedure in Step 2 of Example 1. The crude product was purified by preparative reversed-phase HPLC and dried using lyophilization. ESI-MS m / z 362 (MH) +. EXAMPLE 56: (R) -3- (2- (3-aminocyclobutyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic acid
[0367] [00367] To 2- (3-oxocyclobutyl) methyl acetate (426 mg) and (2,4-dime-toxifenyl) methanamine (501 mg) in a vial was added MeOH (20 mL) and Pd / C (10% , 100 mg). The reaction mixture was stirred under a hydrogen atmosphere overnight. The catalyst was removed by filtration through the Celite buffer. Removal of the solvent provided the crude product (850 mg) which was used directly in the next step without further purification. Step 2: Synthesis of methyl 2- (3- (tert-butoxycarbonyl (2,4-dimethoxybenzyl) amino) cyclobutyl) acetate.
[0368] [00368] To the product from step 1 in DCM (20 ml) was added TEA (0.56 ml) and di-tert-butyl dicarbonate (900 mg). The reaction mixture was stirred at RT overnight. The organic phase was washed with 1 N HCl, water and brine, dried over anhydrous sodium sulfate. Removal of solvents under reduced pressure to provide the product that was purified by flash chromatography (0.75 g). Step 3: Synthesis of 2- (3- (tert-butoxycarbonyl (2,4-dimethoxybenzyl) amino) cyclobutyl) acetic acid.
[0369] [00369] To 2- (3- (tert-butoxycarbonyl (2,4-dimethoxybenzyl) amino) cyclo-butyl) methyl acetate from step 2 in a mixture of MeOH, THF and H2O was added 1N NaOH (10 mL). The resulting reaction mixture was stirred at RT for 2 h. Half of the solvents were removed under reduced pressure and 1N HCl was added to adjust the pH of the solution to 4. The aqueous phase was extracted with EtOAc three times. The combined organic phase was then dried and concentrated in vacuo to provide the acid (0.67 g). Step 4: Synthesis of 3 - ((2R) -2- (2- (3- (tert-butoxycarbonylamino) cyclobutyl) acetamido) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora -tricycle [6.1, 1.02.6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate.
[0370] [00370] Prepared from the 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-ora-tricycle) tert-butyl ester [6,1,1,0 2,6 ] dec-4-ylmethyl) -benzoic acid and 2- (3- (tert-butoxycarbonyl (2,4-dimethoxybenzyl) amino) cyclobutyl) acetic acid after the procedure described in Step 1 of Example 1. The crude product was purified by silica gel scintillation chromatography (hexane / EtOAc, 2: 1 to 1: 2). ESI-MS m / z 641.1 (MH) +. Step 5. Synthesis of (R) -3- (2- (3- (2-aminoethylamino) cyclobutyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2 ] oxaborinin-8-carboxylic.
[0371] [00371] Prepared from 3 - ((2R) -2- (2- (3- (tert-butoxycarbonyl-mino) cyclobutyl) acetamido) -2- (2,9,9-trimethyl-3,5-dioxa -4-bora-tricycle [6,1,1,02'6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 319 (MH) +. EXAMPLE 57: (R) -3- (2- (3- (3- (2-aminoethyl) -1- (2- (3- (2-aminoethyl) ureido) ethyl) ureido) cyclobutyl) acetamido) -2 -hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0372] [00372] Prepared from 3 - ((R) -2- (2- (3- (tert-butoxycarbonylamino) ethyl) amino) cyclobutyl) acetamido) -2 (2,9,9 -tri-methyl-3,5-dioxa-4-bora-tricycle [6,1,1,026] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate (Step 4 of Example 55) and 4 N HCl after the procedure described in Step 1 of Example 53. Step 2: Synthesis of (R) -3- (2- (3- (3- (2-aminoethyl) -1- (2- (3- (2-amino-ethyl) ureido) ethyl) ureido) cyclobutyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0373] [00373] To the 1,1'-carbonyldiimidazole (180 mg) in DCM (5 ml) was added tert-butyl 2-aminoethylcarbamate (160 mg). The resulting reaction mixture was stirred at RT for 1 h. A part of this solution (1 mL) was then added to 3 - ((2R) -2- (2- (3- (2-aminoethylamino) cyclobutyl) acetamido) -2- (2,9,9-trimethyl-3 , 5-dioxa-4-bora-tricycle [6,1,1,026] dec-4-yl) ethyl) -2-methoxybenzoic from step 1 (20 mg) in DMF and TEA (2 eq) in a different bottle. The reaction mixture was stirred overnight. Water was added and extracted with EtOAc. The combined organic phases were dried and concentrated in vacuo to provide the product which was used directly in the next step. Step 3: Synthesis of (R) -3- (2- (3- (3- (2-aminoethyl) -1- (2- (3- (2-amino-ethyl) ureido) ethyl) ureido) cyclobutyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0374] [00374] Prepared from (R) -3- (2- (3- (3- (2-aminoethyl) -1- (2- (3- (2-aminoethyl) ureido) ethyl) ureido) cyclobutyl acid) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by Preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 534 (MH) +. EXAMPLE 58: (R) -3- (2- (3- (3- (2-aminoethyl) ureido) cyclobutyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e ] [1,2] oxaborinin-8-carboxylic
[0375] [00375] Synthesis of (R) -3- (2- (3- (3- (2-aminoethyl) ureido) cyclobutyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e ] [1,2] oxaborinin-8-carboxylic.
[0376] [00376] Prepared from 3 - ((2R) -2- (2- (3- (tert-butoxycarbonylamino) cyclobutyl) acetamido) -2- (2,9,9-trimethyl-3,5-dioxa -4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate (Step 4 of Example 56) after the procedure described in Example 57. The product crude was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 405 (MH) +. EXAMPLE 59: (R) -3- (2- (trans-4- (3- (2-aminoethyl) ureido) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carboxylic
[0377] [00377] Synthesis of (R) -3- (2- (trans-4- (3- (2-aminoethyl) ureido) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H acid -benzo [e] [1,2] oxaborin-8-carboxylic.
[0378] [00378] Prepared from 3 - ((2R) -2- (2- (trans-4-aminocyclohexyl) acetamido) -2- (2,9,9-trimethyl-3,5-dioxa-4) acid -bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-methoxybenzoic (Step 1 of Example 53) after the procedure described in Example 57. The crude product was purified by preparative HPLC reverse phase and dried using lyophilization. ESI-MS m / z 433 (MH) +. EXAMPLE 60: (R) -3- (2- (trans-4- (3-aminopropylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1 , 2] oxaborinin-8-carboxylic
[0379] [00379] The title compound (isolated as a 4: 1 mixture of trans: cis isomers) was prepared using essentially the same procedure used in Example 48, Step 1, except that using tert-butyl N- (3-aminopropyl) carbamate instead of 2- (N- [2- (amino) -ethyl] -amino) -pyridine. Step 2: Synthesis of ethyl 2- [4- [tert-butoxycarbonyl- [3- (tert-butoxycarbonyl-mino) propyl] amino] cyclohexyl] acetate.
[0380] [00380] The title compound (isolated as a 4: 1 mixture of trans: cis isomers) was prepared using essentially the same procedure used in Example 48, Step 2, except that using ethyl 2- [4- [3- (tert -butoxycarbonylamino) propylamino] cyclohexyl] tert-butyl acetate instead of 2- [4- [2- (2-pyridylamino) ethylamino] cyclohexyl] ethyl acetate. Step 3. Synthesis of trans-2- [4- [tert-butoxycarbonyl- [3- (tert-buto-xicarbonylamino) propyl] amino] cyclohexyl] acetic acid.
[0381] [00381] To a solution of 2- [4- [tert-butoxycarbonyl- [3- (tert-butoxy-carbonylamino) propyl] amino] cyclohexyl] ethyl acetate (1.53 g, 3.47 mmol) in methanol (3 ml) and THF (3 ml) NaOH (7.5 ml, 1M aqueous) was added. The resulting solution was stirred for 2.75 hours, acidified with HCl (8 ml, 1M aqueous). This mixture was extracted with ethyl acetate, washed with brine, dried over magnesium sulfate and concentrated. The residue was taken up in ether (5 ml). To this solution was added (-) α-methyl-benzylamine (428 μl, 347 mmol) and the resulting solution left to stand overnight. The crystalline mass was filtered, washed with ether and the collected solid recrystallized from isopropanol / ether to provide 1.1 g of solid. This material was suspended with ethyl acetate, washed with 1 M aqueous HCl, followed by brine, dried over magnesium sulfate and concentrated to provide the title compound. Step 4: Synthesis of acid (+) pinanediolate [1 - [[2- [4- [tert-butoxycarbonyl- [3- (tert-butoxycarbonylamino) propyl] amino] cyclohexyl] acetyl] amino] -2 - (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic.
[0382] The title compound was prepared using essentially the same procedure as described in Example 50; step 9, except that using trans-2- [4- [tert-butoxycarbonyl- [3- (tert-butoxycarbonylamino) propyl] amino] cyclohexyl] acetic acid instead of 2- [4-trans- [tert- butoxycarbonyl- [2- (tert-butoxycarbonylamino) ethyl] amino] cyclohexyl] acetic. Step 5: Synthesis of (R) -3- (2- (trans-4- (3-aminopropylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0383] The title compound was prepared using essentially the same procedure as described in Example 49; step 7 except that using [1 - [[2- [4- [tert-butoxycarbonyl- [3- (tert-butoxycarbonylamino) propyl] amino] cyclohexyl] ace-til] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic acid instead of acid (+) pinanediolate diester [(1R) -1 - [[2- [4- [tert-butoxycarbonyl- [ 2- (methanesulfonamido) ethyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic. EXAMPLE 61: (R) -2-hydroxy-3- (2- (trans-4- (2- (2-hydroxyethylamino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0384] [00384] To (R) -3- (2- (trans-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [ 1.2] oxaborinin-8-carbo-xylic from Example 15 (92 mg) in MeOH (2 ml) was added TEA (70 μl), acetic acid (30 μl), 2- (tert-butyldimethylsilyloxy) acetaldehyde (35 mg ) and sodium triacetoxyborohydride (212 mg). The reaction mixture was stirred overnight at RT. Solvent was removed under reduced pressure and the product was carried on to the next step without further purification. Step 2: Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (2- (2-hydroxyethyl-mino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro -2H-benzo [e] [1,2] oxa-borinine-8-carboxylic
[0385] [00385] To the compound of step 1 was added a mixture of TFA (2 ml) and H2O (0.2 ml). The resulting reaction mixture was stirred at RT for 2 h. The solvents were then removed in vacuo and the residue purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 434 (MH) +. EXAMPLE 62: (R) -3- (2- (trans-4- (2 - ((S) -2-aminopropylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro -2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0386] [00386] To (R) -3- (2- (trans-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [ 1.2] oxaborinin-8-carbo-xylic from Example 15 (92 mg) in MeOH (2 mL) TEA (70 μΙ), acetic acid (30 μl), 1-oxopropan-2-ylcarbamate (S) was added -tert-butyl (86 mg) and sodium triacetoxyborohydride (212 mg). The reaction mixture was stirred overnight at RT. The solvent was removed and the product was carried on to the next step without further purification. Step 2: Synthesis of (R) -3- (2- (trans-4- (2 - ((S) -2-aminopropylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4 acid -dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0387] [00387] To the compound of step 1, 3N HCl (2 ml) was added and the resulting reaction mixture was heated to reflux for 1 h. The solvents were then removed in vacuo and the residue purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 447 (MH) +. EXAMPLE 63: (R) -3- (2- (trans-4 - ((2-aminoethyl) (methyl) amino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- acid benzo [e] [1,2] oxaborinin-8-carboxylic.
[0388] The title compound (isolated as a 6.5: 1 mixture of trans: cis isomers) was prepared using essentially the same procedure used in Example 48, Step 1, except that using N- (3-aminoethyl) carbamate tert-butyl instead of 2- (N- [2- (amino) -ethyl] -amino) -pyridine. Step 2: Synthesis of ethyl 2- [4- [2- (tert-butoxycarbonylamino) ethyl-methyl-amino] cyclohexyl] acetate.
[0389] [00389] To a solution of 2- [4- [2- (tert-butoxycarbonylamino) ethylamino] cyclohexyl] ethyl acetate (326 mg, 1 mmol) in dichloromethane (4 mL) was added formalin (97 μΙ , 1.2 mmol) followed by acetic acid (60 μl, 1 mmol) and sodium triacetoxyborohydride (255 mg, 1.2 mmol). The resulting cloudy solution was stirred for 19 hours. To these mixture was added sodium carbonate (2 ml, saturated aqueous solution). The mixture was diluted with ethyl acetate and separated. The organic extract was washed with brine, dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (10 g silica; eluted with 2 to 20% methanol in dichloromethane) to provide the title compound. Step 3: Synthesis of 2- [4- [2- (tert-butoxycarbonylamino) ethyl-methyl-amino] cyclohexyl] acetic acid.
[0390] [00390] To a solution of 2- [4- [2- (tert-butoxycarbonylamino) ethyl-methyl-amino] cyclohexyl] ethyl acetate (231 mg, 0.67 mmol) in methanol (2 mL) and THF (2 ml) NaOH (1 ml, 2 M aqueous solution) was added. The resulting solution was stirred for 2.75 hours, then concentrated under reduced pressure to approximately% of the original volume. The residue was acidified with HCl (3 ml, 1 M aqueous solution). This solution was purified directly by reverse phase chromatography on silica (30 g of C18 silica; eluted with 5-100% acetonoitrile / water / 0.1% TFA) to provide the title compound. Step 4: Synthesis of acid (+) pinanediolate diester [1 - [[2- [4- [2- (tert-butoxycarbonylamino) ethyl-methyl-amino] cyclohexyl] acetyl] amino] -2- (3 -tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic.
[0391] The title compound was prepared using essentially the same procedure as described in Example 50; step 9, except that using 2- [4- [2- (tert-butoxycarbonylamino) ethyl-methyl-amino] cyclohexyl] acetic acid instead of 2- [4-trans- [tert-butoxycarbonyl- [2- (tert-butoxycarbonylamino) ethyl] amino] cyclohexyl] acetic. Step 6: Synthesis of (R) -3- (2- (trans-4 - ((2-aminoethyl) (methyl) amino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro- 2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0392] The title compound was prepared using essentially the same procedure as described in Example 49; step 7 except using [1 - [[2- [4- [2- (tert-butoxycarbonylamino) ethyl-methyl-amino] cyclohexyl] acetyl] amino] -2- (3) (+) pinanediolate diester -tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic acid instead of acid (+) pinanediolate diester [(1R) -1 - [[2- [4- [tert-butoxycarbonyl- [2- (methanes- sulfonamido) ethyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic. EXAMPLE 64: (R) -2-hydroxy-3- (3-hydroxy-2- (trans-4- (2- (methylmino) ethylamino) cyclohexyl) propanamido) -3,4-dihydro acid -2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0393] [00393] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,0 26] dec -4-ylmethyl) -benzoic and 2- (trans-4- (tert-butoxycarbonyl (2- (tert-butoxycarbonyl (methyl) amino) ethyl) amino) cyclohexyl) acetic acid after the procedure described in step 1 of the Example 1. The crude product was purified by silica gel scintillation chromatography (hexane / EtOAc, 2: 1 to 1: 2). Step 2. Synthesis of 3 - ((2R) -2- (2- (trans-4- (tert-butoxycarbonyl (2- (tert-butoxycarbonyl (methyl) amino) ethyl) amino) cyclohexyl) -3-hydroxypropanamido ) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-tert-methoxybenzoate butyl.
[0394] [00394] Ao 3 - ((2R) -2- (2- (trans-4- (tert-butoxycarbonyl (2- (tert-butoxycarbonyl (methyl) amino) ethyl) amino) cyclohexyl) acetamido) - 2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) -2-tert-butyl methoxybenzoate ( 110 mg) in THF (5 ml) LDA (2 M in benzene, 160 μl) was added at -76 ° C. The reaction mixture was stirred at the same temperature for 30 min before formaldehyde (20 mg) was added. The reaction mixture was allowed to warm to RT and stirred for 4 h. Brine was added and extracted with EtOAc. The organic phase was dried and concentrated to provide the crude product which was used in the next step without further purification. Step 3: Synthesis of (R) -2-hydroxy-3- (3-hydroxy-2- (trans-4- (2- (methylamino) ethylamino) cyclohexyl) propanamido) -3,4-dihydro -2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0395] [00395] Prepared from 3 - ((2R) -2- (2- (trans-4- (tert-butoxycarbonyl (2- (tert-butoxycarbonyl (methyl) amino) ethyl) amino) cyclohexyl) -3-hydro-xipropanamido) -2- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) - Tert-butyl 2-methoxybenzoate and BCl3 after the procedure described in Step 2 of Example 1. The crude product was purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 434 (MH) +. EXAMPLE 65: (R) -3- (2- (trans-4 - ((R) -2-amino-3-hydroxypropyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-di- hydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0396] [00396] To a solution of 2- [4- (tert-butoxycarbonylamino) cyclohexyl] acetic acid (1.0 g, 4 mmol) in DMF (10 mL) was added K2CO3 (600 mg, 4 mmol) followed by benzyl bromide (0.5 mL, 4.2 mmol). The resulting mixture was stirred for 16.75 hours, diluted with ether, washed with water (2X) followed by brine, dried over magnesium sulfate and concentrated. The residue was taken up in dichloromethane (12 ml). To this solution was added TFA (3 ml). The resulting solution was stirred for 2.5 hours then concentrated under reduced pressure. The residue was taken up in ethyl acetate, washed with saturated sodium bicarbonate solution followed by brine, dried over magnesium sulfate and concentrated under reduced pressure to provide the title compound as a white solid. Step 2: Synthesis of (3R) -4 - [[[4- (2-benzyloxy-2-oxo-ethyl) cyclohexyl] -tert-butoxycarbonyl-amino] methyl] -2,2-dimethyl-oxazolidine-3 tert-butyl carboxylate.
[0397] [00397] To a solution of tert-butyl (3R) -4-formyl-2,2-dimethyl-oxazolidine-3-carboxylate (460 mg, 2 mmol) in dichloromethane (6 mL) was added 2- [trans- Benzyl 4- (amino) -cyclohexyl] acetate (494 mg, 2 mmol) followed by sodium triacetoxyborohydride (636 mg, 3 mmol) and acetic acid (120 μl, 2 mmol). The resulting solution was stirred for 16.75 hours. To this solution was added sodium carbonate (aqueous saturated solution). The mixture was diluted with ethyl acetate, washed with brine, dried over magnesium sulfate and concentrated. The residue was taken up in dichloromethane (6 ml). To this solution was added di-tert-butyl dicarbonate (654 mg, 3 mmol) followed by triethylamine (460 μl 3.3 mmol). This solution was stirred for 2 hours then diluted with ether, washed with brine, dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (25 g silica; eluted with 10 to 40% ethyl acetate in hexanes) to provide the title compound as a white solid. Step 3: Synthesis of 2- [4- [tert-butoxycarbonyl - [[((3R) -3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4-yl] methyl] amino] cyclohexyl] acetic acid.
[0398] [00398] To a solution of (3R) -4 - [[[4- (2-benzyloxy-2-oxo-ethyl) cyclohexyl] -tert-butoxycarbonyl-amino] methyl] -2,2-dimethyl-oxazolidine -3-tert-butyl carboxylate (520 mg, 0.928 mmol) in ethyl acetate (4 mL) was added palladium on carbon (56 mg, 10% palladium on dry carbon powder). The mixture was subjected to the gas removal treatment, flushed with hydrogen gas and stirred under this atmosphere for 20 minutes. The system was again subjected to gas removal treatment, then flushed with argon. This mixture was diluted with dichloromethane, filtered through celite and concentrated under reduced pressure. The residue was triturated with hexane to provide the title compound as a white solid. Step 4: Synthesis of acid (+) pinanediolate diester [1 - [[2- [4- [tert-butoxycarbonyl - [[(3R) -3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4- yl] methyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic.
[0399] The title compound was prepared using essentially the same procedure as described in Example 50; step 9, except that using 2- [4- [tert-butoxycarbonyl - [[((3R) -3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4-yl] methyl] amino] cyclohexyl] acetic instead of 2- [4-trans- [tert-butoxycarbonyl- [2- (tert-butoxycarbonylamino) ethyl] amino] cyclohexyl] acetic acid. Step 5: Synthesis of (R) -3- (2- (trans-4 - ((R) -2-amino-3-hydroxy-propylamino) cyclohexyl) acetamido) -2-hydroxy-3,4- dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0400] The title compound was prepared using essentially the same procedure as described in Example 49; step 7, except that using acid (+) pinanediolate diester [1 - [[2- [4- [tert-butoxycarbonyl - [[(3R) -3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4 -yl] methyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic acid instead of acid (+) pinanediolate diester [(1R) - 1 - [[2- [4- [ferc-butoxycarbonyl- [2- (methanesulfonamido-) ethyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl ] boronic. EXAMPLE 66: (R) -2-hydroxy-3- (2- (trans-4- (2- (methylthio) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carboxylic
[0401] [00401] The title compound was prepared using essentially the same procedure used in Example 48, Step 1 except that using 2- (methylthio) -ethylamine instead of 2- (N- [2- (amino) -ethyl] -amino ) -pyridine. Step 2: Synthesis of ethyl 2- [4- [tert-butoxycarbonyl (2-methylsulfanylethyl) amino] cyclohexyl] acetate.
[0402] The title compound (isolated as a 6.8: 1 mixture of trans: cis isomers) was prepared using essentially the same procedure used in Example 48, Step 2, except that using 2- [4- (2-methylsulfanylethylamino) ) ethyl cyclohexyl] ethyl acetate instead of ethyl 2- [4- [2- (2-pyridylamino) ethylamino] cyclohexyl] acetate. Step 3: Synthesis of 2- [4- [tert-butoxycarbonyl (2-methylsulfanylethyl) amino] cyclohexyl] acetic acid.
[0403] [00403] To a solution of 2- [4- [tert-butoxycarbonyl (2-methylsulfa-ethylethyl) amino] cyclohexyl] ethyl acetate (2.11 g, 5.89 mmol) in methanol (10 mL) and THF (10 ml) NaOH (10 ml, 1M aqueous) was added. The resulting solution was stirred for 3.75 hours, then acidified with HCl (2M, aqueous to pH 2). This mixture was extracted with ethyl acetate, washed with brine, dried over magnesium sulfate and concentrated. The residue was purified by chromatography on silica (25 g silica; eluted with 20 - 60% ethyl acetate in hexanes) to provide the title compound as a 6.9: 1 mixture of trans to cis isomers. Step 4: Synthesis of [2- (3-tert-) acid (+) pinanediolate diester butoxycarbonyl-2-methoxy-phenyl) -1 - [[2- [4- [tert-butoxycarbonyl (methylsulfanylmethyl) amino] cyclohexyl] acetyl] amino] ethyl] boronic.
[0404] The title compound was prepared using essentially the same procedure as described in Example 50; step 9, except that using 2- [4- [tert-butoxycarbonyl (2-methylsulfanyl-til) amino] cyclohexyl] acetic acid instead of 2- [4-trans- [tert-butoxy-bonyl- [2 - (tert-butoxycarbonylamino) ethyl] amino] cyclohexyl] acetic. Step 5: Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (2- (methylthio) ethylamine) cyclohexyl) acetamido) -3,4-dihydro-2H -benzo [e] [1,2] oxaborinin-8-carboxylic.
[0405] The title compound was prepared using essentially the same procedure as described in Example 49; step 7, except that using [2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) -1 - [[2- [4- [tert-butoxycarbonyl (methylsulfanyl-methyl) amino] diester ] cyclohexyl] acetyl] amino] ethyl] boronic acid instead of acid (+) pinanediolate diester [(1R) -1 - [[2- [4- [tert-butoxycarbonyl- [2- (meta-ourulfonamido) ethyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic. EXAMPLE 67: (R) -3- (2- (trans-4 - ((S) -2-amino-3-hydroxypropyl- mino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic
[0406] The title compound was prepared using essentially the same procedure as used in Example 65; step 2, except using tert-butyl (3S) -4-formyl-2,2-dimethyl-oxazolidine-3-carboxylate instead of (3R) -4-formyl-2,2-dimethyl-oxazolidine-3- tert-butyl carbo-xylate. Step 2: Synthesis of 2- [4- [tert-butoxycarbonyl - [[(3S) -3-tert-butoxy-carbonyl-2,2-dimethyl-oxazolidin-4-yl] methyl] amino] cyclohexyl]] acetic.
[0407] The title compound was prepared using essentially the same procedure used in Example 65; step 3, except that using (3S) -4 - [[[4- (2-benzyloxy-2-oxo-ethyl) cyclohexyl] -tert-butoxycarbonyl-amino] methyl] -2,2-dimethyl-oxazolidine- Tert-butyl 3-carboxylate instead of (3R) -4 - [[[4- (2-benzyloxy-2-oxo-ethyl) cyclohexyl] -tert-butoxycarbonyl-amino] methyl] -2,2- tert-butyl dimethyl-oxazolidine-3-carboxylate. Step 3: Synthesis of acid (+) pinanediolate diester [1 - [[2- [4- [tert-butoxycarbonyl - [[(3S) -3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4- il] methyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic
[0408] The title compound was prepared using essentially the same procedure as described in Example 50; step 9, except that using 2- [4- [tert-butoxycarbonyl - [[((3S) -3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4-yl] methyl] amino] cyclohexyl] acetic acid instead of 2- [4-trans- [tert-butoxycarbonyl- [2- (tert-butoxycarbo-nylamino) ethyl] amino] cyclohexyl] acetic acid. Step 4: Synthesis of (R) -3- (2- (trans-4 - ((S) -2-amino-3-hydroxypropylamino) cyclo-exyl) acetamido) -2-hydroxy-3,4- dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0409] The title compound was prepared using essentially the same procedure as described in Example 49; step 7, except that using acid (+) pinanediolate diester [1 - [[2- [4- [tert-butoxycarbonyl - [[(3S) -3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4 -yl] methyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic acid instead of acid (+) pinanediolate diester [(1R) - 1 - [[2- [4- [tert-butoxycarbonyl- [2- (methanesulfonamido) ethyl] amino] cyclohexyl] acetyl] amino] -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) ethyl] boronic. EXAMPLE 68: (R) -3- (2- (trans-4- (2-aminoacetamido) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1 , 2] oxaborinin-8-carbo-xyl
[0410] [00410] To 2- (tert-butoxycarbonylamino) acetic acid (175 mg) in a vial was added HATU (380 mg) and N-methyl morpholine (0.56 ml). After stirring at RT for 1 h, this solution was added to 3 - ((2R) -2- (2- (trans-4-aminocyclohexyl) acetamido) -2- (2,9,9-trimethyl-3 , 5-dioxa-4-bora-tricycle [6,1,1,0 2,6] dec-4-yl) ethyl) -2-methoxybenzoic from step 1 of Example 53 (110 mg) in DMF (3 ml) . The resulting reaction mixture was stirred at RT overnight. Water was added to the reaction mixture and extracted with EtOAc. The organic phase was dried and concentrated in vacuo to provide the crude product which was used in the next step without further purification. Step 2: Synthesis of (R) -3- (2- (trans-4- (2-aminoacetamido) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0411] [00411] Prepared from 3 - ((2R) -2- (2- (trans-4- (2- (tert-butoxycarbonylamino) acetamido) cyclohexyl) acetamido) -2- (2,9,9 -trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,026] dec-4-yl) ethyl) -2-methoxybenzoic after the procedure described in Step 2 of Example 62. The product was purified by HPLC preparative phase and dried using lyophilization. ESI-MS m / z 404 (MH) +. EXAMPLE 69: (R) -3- (2- (trans-4- (bis ((1H-imidazol-2-yl) methyl) amino) cyclohexyl) acetamido) -2-hydroxy-3,4-di -hydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0412] [00412] Synthesis of (R) -3- (2- (trans-4- (bis ((1H-imidazol-2-yl) methyl) amino) cyclohexyl) acetamido) -2-hydroxy-3,4 acid -dihydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0413] [00413] Prepared from (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1, 2] oxaborinin-8-carboxylic (Example 6) and 1H-imidazole-2-carbaldehyde after the procedure of Example 27. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 507 (MH) +. EXAMPLE 70: (R) -3- (2- (trans-4 - (((1H-imidazol-5-yl) methylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H acid -benzo [e] [1,2] oxaborinin-8-carboxylic.
[0414] [00414] Synthesis of (R) -3- (2- (trans-4 - ((1H-imidazol-5-yl) methyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-di acid -hydro-2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0415] [00415] Prepared from (R) -3- (2- (trans-4-aminocyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1, 2] oxaborinin-8-carboxylic (Example 6) and 1H-imidazole-5-carbaldehyde after the procedure of Example 27. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 427 (MH) +. EXAMPLE 71: (R) -2-hydroxy-3- (2- (trans-4- (2- (isopropylamino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H-benzo [e ] [1,2] oxaborinin-8-carboxylic.
[0416] [00416] Synthesis of (R) -2-hydroxy-3- (2- (trans-4- (2- (isopropyl-mino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H -benzo [e] [1,2] oxaborinin-8-carboxylic.
[0417] [00417] To (R) -3- (2- (trans-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [ 1.2] oxaborinin-8-carboxylic acid from Example 15 (92 mg) in MeOH (2 ml) was added TEA (70 μl), acetic acid (30 μl), acetone (0.1 ml) and sodium triacetoxyborohydride (212 mg). The reaction mixture was stirred overnight at RT. The solvent was removed and the product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 432 (MH) +. EXAMPLE 72: (R) -2-hydroxy-3- (2-trans-4- (2- (pyrimidin-2-ylamino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro-2H- benzo [e] [1,2] oxaborinin-8-carboxylic.
[0418] [00418] Synthesis of (R) -2-hydroxy-3- (2-trans-4- (2- (pyrimidin-2-ylamino) ethylamino) cyclohexyl) acetamido) -3,4-dihydro- 2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0419] [00419] To (R) -3- (2- (trans-4- (2-aminoethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e] [ 1.2] oxaborinin-8-carboxylic acid from Example 15 (46 mg) in MeOH (2 ml) was added TEA (70 μl) and 2-chloropyrimidine (25 mg). The reaction mixture was stirred at 70 ° C overnight. The solvent was removed and the product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 468 (MH) +. EXAMPLE 73: (R) -3- (2- (trans-4- (2- (cyclopentylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e ] [1,2] oxaborinin-8-carboxylic.
[0420] [00420] Synthesis of (R) -3- (2- (trans-4- (2- (cyclopentylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carboxylic.
[0421] [00421] Prepared from (R) -3- (2- (trans-4- (2-aminoethyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- benzo [e] [1,2] oxa-borinine-8-carboxylic and cyclopentanone after the procedure described in Example 71. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 458 (MH) +. EXAMPLE 74: (R) -3- (2- (trans-4- (2- (cyclopropylmethylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e ] [1,2] oxaborinin-8-carboxylic.
[0422] [00422] Synthesis of (R) -3- (2- (trans-4- (2- (cyclopropylmethylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carboxylic.
[0423] [00423] Prepared from (R) -3- (2- (trans-4- (2-aminoethyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- benzo [e] [1,2] oxa-borinine-8-carboxylic and cyclopropanecarbaldehyde after the procedure described in Example 71. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 444 (MH) +. EXAMPLE 75: (R) -3- (2- (trans-4- (2- (bis (cyclopropylmethyl) amino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H acid -benzo [e] [1,2] oxaborinin-8-carboxylic.
[0424] [00424] Synthesis of (R) -3- (2- (trans-4- (2- (bis (cyclopropylmethyl) amino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro acid -2H-benzo [e] [1,2] oxaborinin-8-carboxylic.
[0425] [00425] Prepared from (R) -3- (2- (trans-4- (2-aminoethyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- benzo [e] [1,2] oxa-borinine-8-carboxylic and cyclopropanecarbaldehyde after the procedure described in Example 71. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 498 (MH) +. EXAMPLE 76: (R) -3- (2- (trans-4- (1,3-diaminopropyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carbo-xyl.
[0426] [00426] Synthesis of 2- (trans-4- (2,2,12,12-tetramethyl-4,10-dioxo-3,11-doxa-5,9-diazatridecan-6-yl) cyclohexyl) acid acetic. Step 1: Synthesis of 2- (1,4-dioxaspas [4,5] decan-8-yl) ethyl acetate.
[0427] [00427] To a cooled (0 ° C) suspension of NaH (60%, 4.4 g, 110 mmol) in THF (200 mL) was added triethyl phosphonoacetate (25.16 g, 110 mmol) at a rate of to produce a smooth progressive gas development. After completing the addition, the homogeneous solution was stirred for 30 min. To this solution was added 1,4-cyclohexanedione monoethylene ketal (15.62 g, 100 mmol) in THF (40 ml) for 10 min. After completing the addition, the ice bath was removed and stirring was continued for 3 h. The reaction was quenched by the addition of saturated aqueous NH4Cl, extracted with EtOAc, washed with brine, dried over Na2SO4. Filtration and evaporation to dryness provided the crude product which was used in the next step without further purification.
[0428] [00428] The above crude product (25.3 g, 100 mmol) was dissolved in MeOH (80 ml) and 10% Pd / C (1 g) added. The resulting mixture was hydrogenated at 35 psi for 3 h. After filtration and evaporation, the residue was purified by scintillating column chromatography (eluent: 20% EtOAc in 30% hexanes) which provided the product as a colorless oil (20 g, 87%). Step 2: Synthesis of 2- (1,4-dioxaspospiro [4,5] decan-8-yl) ethanol.
[0429] [00429] To a solution of 2- (1,4-dioxaspospiro [4,5] decan-8-yl) ethyl acetate from step 1 (3.31 g, 14.5 mmol) in Et2O (80 mL) at 0 ° C under N2 LiAlH4 (1M in THF, 13.66 mL, 13.66 mmol) was added in 15 min. The resulting mixture was stirred at 0 ° C for a further 20 min and quenched by the addition of saturated aqueous NH4Cl solution, extracted with EtOAc, dried over Na2SO4. Filtration and evaporation to dryness gave the alcohol as a white solid (2.76 g, 100%). LC / MC: 187.1 (MH) +. Step 3: Synthesis of 8- (2- (benzyloxy) ethyl) -1,4-dioxaspas [4,5] decane.
[0430] [00430] To a pre-cooled (0 ° C) suspension of NaH (60% mineral, 0.44 g, 11 mmol) in THF (20 mL) was added 2- (1,4-dioxaspospiro [4,5 ] decan-8-yl) ethanol (1.86 g, 10 mmol) in THF (10 mL). The resulting solution was stirred at 0 ° C for 10 min, and allowed to warm to RT, and stirred for 1 h. To the above mixture was added benzyl bromide (1.78 g, 15 mmol) and the resulting mixture was stirred at RT overnight. The reaction was quenched by the addition of saturated aqueous NH4Cl solution, extracted with EtOAc, washed with brine, dried over Na2SO4. Purification by flashing column chromatography (eluent: 20% EtOAc in 30% hexanes) gave the product (2.3 g, 83%). LC / MC: 277.1 (MH) +. Step 4: Synthesis of 4- (2- (benzyloxy) ethyl) cyclohexanone.
[0431] [00431] A solution of 8- (2- (benzyloxy) ethyl) -1,4-dioxaspospiro [4,5] decane (2.30 g, 8.32 mmol) in acetonitrile (18 mL) was added a solution of 6 N HCl and the resulting solution was stirred at RT for 2 h. After removing acetonitrile through evaporation, the residue was neutralized by solid NaHCO3, extracted with EtOAc, dried by Na2SO4. Filtration and evaporation to dryness gave the ketone as a white solid (1.84 g, 95.5%). Step 5: Synthesis of ((2- (4-methylenocyclohexyl) ethoxy) methyl) benzene.
[0432] [00432] To a cooled (-78 ° C) mixture of Ph3PCH2Br (5.42 g, 14.88 mmol) in THF (25 mL) was added COtBu (1M in THF, 17.05 mL, 17.05 mmol) by drip under N2. The resulting mixture was stirred at 0 ° C for 1 h and heated to RT for an additional 1.5 h. The reaction mixture was then cooled to -40 ° C, a solution of 4- (2- (benzyloxy) ethyl) cyclohexanone (1.8 g, 7.75 mmol) in THF (15 mL) was added by dropping. The mixture was then stirred at RT overnight and quenched by brine, extracted with EtOAc, dried over Na2SO4. Purification by flashing column chromatography (eluent: 20% EtOAc in 30% hexanes) gave the title product (1.77 g, 99%). LC / MC: 231.1 (MH) +. Step 6: Synthesis of (trans-4- (2- (benzyloxy) ethyl) cyclohexyl) methanol
[0433] [00433] To a cooled (0 ° C) solution of ((2- (4-methylenocyclohexyl) ethoxy) methyl) benzene (9.68 g, 41.66 mmol) in THF (200 mL) was added B2H6 Me2S (2M in THF, 41.66 mL, 83.32 mmol) under N2. After being stirred at 0 ° C for 2 h and at RT for another 2 h, the solution was cooled to 0 ° C and a mixture of 3 M aqueous NaOH solution (34 mL) and 30% hydrogen peroxide solution (34) was added. mL) by dripping. The resulting mixture was stirred at 0 ° C for 1 h and at RT for 1.5 h. Aqueous preparation and purification by sparkling column chromatography (eluent: 40% EtOAc in hexanes) provided the product as a yellow oil (7.04 g, 68%). LC / MC: 249.1 (MH) +. Step 7: Synthesis of (trans-4- (2- (benzyloxy) ethyl) cyclohexanecarbaldehyde.
[0434] [00434] A solution of DMSO (2.20 mL, 31 mmol) in DCM (45 mL) was added by dripping to a pre-cooled (-78 ° C) solution of oxalyl chloride (2.9 mL, 33, 8 mmol) in DCM (45 mL) under N2. After stirring at -78 ° C for 10 min, a solution of (trans-4- (2- (benzyloxy) ethyl) cyclohexyl) methanol (7.0 g, 28.18 mmol) in DCM (45 mL) was added by dripping. The resulting solution was stirred at -78 ° C for 15 min, and TEA (23.6 ml, 169.1 mmol) was added. The solution was stirred at -78 ° C for 15 min and at RT for 20 min. The reaction was diluted with DCM, washed with 1 M HCl solution and brine, dried over Na2SO4. Purification by flashing column chromatography (eluent: 20% EtOAc in hexanes) gave the aldehyde product as a yellow oil (6.86 g, 98.8%). Step 8: Synthesis of (R, E) -N - ((trans-4- (2- (benzyloxy) ethyl) cyclohexyl) methylene) -2-methylpropane-2-sulfinamide.
[0435] [00435] To a solution of (trans-4- (2- (benzyloxy) ethyl) cyclohexane-carbaldehyde (6.45 g, 26.18 mmol) and (R) - (+) - tert-butylsulfinamide (3 , 49 g, 28.8 mmol) in THF (66 mL) titanium (IV) ethoxide (8.78 mL, 41.89 mmol) was added under N 2. The resulting solution was stirred at RT for 20 h and quickly cooled by addition of saturated NaHCO3 solution by dripping The mixture was vigorously stirred for 30 min and filtered through a Celite buffer, concentration. Purification by sparkling column chromatography (eluent: 20% EtOAc in 30% hexanes) provided the product of titer (8.38 g, 92%) LC / MC: 350.1 (MH) +. Step 9: Synthesis of (R) -N- (1- (trans-4- (2- (benzyloxy) ethyl) cyclohexyl) but-3-enyl) -2-methylpropane-2-sulfinamide.
[0436] [00436] To a cooled (0 ° C) solution of (R, E) -N - ((trans-4- (2- (well-zyloxy) ethyl) cyclohexyl) methylene) -2-methylpropane-2- sulfinamide in DCM (65 ml) allyl magnesium chloride (2 M in THF, 5.87 ml, 11.74 mmol) was added by dropping under N2. The resulting mixture was stirred at 0 ° C for 1 h and quickly cooled by the addition of saturated NH4Cl solution, separated. The aqueous phase was extracted with EtOAc and the combined organic phase was washed with brine, dried over Na2SO4. Purification by flashing column chromatography (eluent: 20% EtOAc in hexanes) gave the product (2.57 g, 100%). LC / MC: 392.1 (MH) +. Step 10: Synthesis of tert-butyl 1 - ((trans-4- (2- (benzyloxy) ethyl) cyclohexyl) but-3-enyl-carbamate
[0437] [00437] To a stirred solution of (R) -N- (1- (trans-4- (2- (benzyloxy) ethyl) cyclohexyl) but-3-enyl) -2-methylpropane-2-sulfinamide (2 , 56 g, 6.5 mmol) in MeOH (3.5 mL) a solution of 4 M HCl in 1,4-dioxane (3.25 mL, 13 mmol) was added. The resulting solution was stirred at RT for 30 min and then concentrated. The residue was dissolved in EtOAc, washed with brine, dried over Na2SO4. Filtration and evaporation to dryness gave the crude product.
[0438] [00438] To a chilled (0 ° C) solution of the above crude product in DCM (40 mL) was added di-tert-butyl dicarbonate (1.75 g, 7.8 mmol) and TEA (3.62 mL) , 26 mmol). The mixture was slowly warmed to RT and stirred at RT for 4 h. The reaction was quenched by the addition of a saturated NH4Cl solution, separated. The aqueous phase was extracted with DCM and the combined DCM extracts were washed with 20% citric acid, brine, dried over Na2SO4. Purification by flashing column chromatography (eluent: 20% EtOAc in hexanes) provided the product as a colorless gel (2.78 g, 40%). LC / MC: 388.1 (MH) +. Step 11: Synthesis of tert-butyl 1- (trans-4- (2- (benzyloxy) ethyl) cyclohexyl) -3-oxo-propylcarbamate.
[0439] [00439] To a solution of tert-butyl 1 - ((trans-4- (2- (benzyloxy) ethyl) cyclohexyl) but-3-enylcarbamate (6.5 mmol) in 1,4-dioxane (118 mL) and water (38 mL) were added N-methylmorpholine-N-oxide (1.52 g, 13 mmol) and OsO4 (4 wt.% with water, 1.4 mL, 0.23 mmol). it was stirred at RT for 18 h and NaIO4 (4.87 g, 22.75 mmol) was added. The resulting mixture was stirred at RT for 4 h and quickly cooled by the addition of saturated aqueous Na2S2O3 solution. was purified by scintillating column chromatography (eluent: 10% EtOAc in 30% hexanes) to provide the aldehyde as a colorless oil (2.36 g, 89.7%) LC / MC: 412.1 (MNa) +. Step 12: Synthesis of tert-butyl 1- (trans-4- (2-hydroxyethyl) cyclohexyl) -3-oxopropyl-carbamate.
[0440] [00440] A solution of the aldehyde from the above step (2.36 g, 6.06 mmol) in MeOH (20 mL) was added 10% Pd / C (0.2 g). The resulting mixture was hydrogenated via an H2 balloon at RT overnight. Filtration and evaporation to dryness gave the product as a colorless gel (1.87 g, 100%). LC / MS: 322.1 (MNa) +. Step 13: Synthesis of 2- (trans-4- (1- (tert-butoxycarbonyl-mino) -3-oxopropyl) cyclohexyl) ethyl acetate.
[0441] [00441] To a solution of 1- (trans-4- (2-hydroxyethyl) cyclohexyl) -3-oxopropylcarbamate of tert-butyl (1.88 g, 6.28 mmol) in DCM (60 mL) DMAP (cat. Quantity), followed by the addition of TEA (2.63 ml, 18.84 mmol) and acetic anhydride (0.89 ml, 9.42 mmol) at 0 ° C under N2. The mixture was stirred at RT for 3 h, and diluted with DCM, cooled quickly by adding aqueous NaHCO3 solution. The organic phase was separated and dried by Na2SO4. Purification by flashing column chromatography (eluent: 40% EtOAc in hexanes) provided the product as a colorless gel (1.90 g, 88.6%). Step 14: Synthesis of 2- (trans-4- (1- (tert-butoxycarbonyl-mino) -3-hydroxypropyl) cyclohexyl) ethyl acetate.
[0442] [00442] To a cooled solution (ethylene glycol + dry ice) of 2- (trans-4- (1- (tert-butoxycarbonylamino) -3-oxopropyl) cyclohexyl) ethyl acetate (1.88 g, 5, 5 mmol) in ethanol (90 mL) NaBH4 (0.212 g, 5.5 mmol) was added. The mixture was stirred at -10 ~ 15 ° C for 10 min and 0 ° C for 10 min, and then quenched by the addition of saturated aqueous NH4G solution (30 ml) and brine (30 ml). After removing the volatile by evaporation, the aqueous residue was extracted with EtOAc, dried over Na2SO4. Purification by flashing column chromatography (eluent: 30% EtOAc in 40% hexanes) provided the alcohol product (1.0 g, 53%). LC / MC: 366.1 (MNa) +. Step 15: Synthesis of 2- (trans-4- (3-azido-1- (tert-butoxycarbonylamino) propyl) cyclohexyl) ethyl acetate
[0443] [00443] To a cooled (0 ° C) solution of 2- (trans-4- (1- (tert-butoxycarbonylamino) -3-hydroxypropyl) cyclohexyl) ethyl acetate (1.0 g, 2.9 mmol ) in DCM (15 ml) TEA (0.81 ml, 5.82 mmol) and methanesulfonyl chloride (0.34 ml, 4.35 mmol) were added by dropping under N2. The mixture was stirred at RT for 3 h and diluted with DCM, washed with aqueous NH4Cl solution, dried over Na2SO4. Purification by flashing column chromatography (eluent: 40% EtOAc in hexanes) provided the mesylate as a yellow oil (0.96 g, 79%). LC / MC: 444.1 (MNa) +.
[0444] [00444] A mixture of mesylate (0.95 g, 2.25 mmol) and NaN3 (1.17 g, 18 mmol) in DMF (25 mL) was heated to 80 ° C overnight. After aqueous preparation, the crude product was purified by scintillating column chromatography (eluent: 30% EtOAc in hexanes) to provide the azide as a yellow oil (0.74 g, 89%). LC / MS: 391.1 (MNa) +. Step 16: Synthesis of 2- (trans-4- (3-amino-1- (tert-butoxycarbonylamino) propyl) cyclohexyl) ethyl acetate.
[0445] [00445] A mixture of the azide from step 15 (0.72 g, 1.95 mmol) and 10% Pd / C (0.1 g) in MeOH (20 mL) was hydrogenated via an H2 balloon at RT for 18 h. Filtration and evaporation to dryness provided the amine in a quantitative yield. LC / MS: 343.1 (MH) +. Step 17: Synthesis of 2- (trans-4- (2,2,12,12-tetramethyl-4,10-dioxo-3,11-doxa-5,9-diazatridecan-6-yl) cyclohexyl acetate ) ethyl.
[0446] [00446] To a cooled (0 ° C) solution of 2- (trans-4- (3-amino-1- (tert-butoxycarbonylamino) propyl) cyclohexyl) ethyl acetate (1.95 mmol) in DCM ( 15 ml) di-tert-butyl dicarbonate (0.53 g, 2.34 mmol) and TEA (1.09 ml, 7.8 mmol) were added. The mixture was slowly heated to RT and stirred at RT overnight. After being quickly cooled through the aqueous NH4Cl solution, the reaction was extracted with DCM, washed with 0.5 N aqueous HCl solution, brine, and dried over Na2SO4. Filtration and evaporation to dryness gave the crude product. LC / MS: 465.1 (MNa) +. Step 18: Synthesis of tert-butyl 1- (4- (2-hydroxyethyl) cyclohexyl) propane-1,3-diildi-carbamate.
[0447] [00447] To an acetate solution of 2- (trans-4- (2,2,12,12-tetra-methyl-4,10-dioxo-3,11-doxa-5,9-diazatridecan-6-yl ) cyclohexyl) ethyl (1.95 mmol) in MeOH (5 mL) K2CO3 (0.135 g, 0.975 mmol) was added and the resulting mixture was stirred at RT for 3 h. The reaction was quenched by the addition of aqueous NH4CL solution. After removing MeOH by evaporation, the residue was extracted with EtOAc, dried over Na2SO4. Filtration and evaporation to dryness gave the product as a white foam (0.71 g, 91%). LC / MS: 423.1 (MNa) +. Step 19: Synthesis of 2- (trans-4- (2,2,12,12-tetramethyl-4,10-dioxo-3,11-dioxide-5,9-diazatridecan-6-yl) acid) cyclohexyl) acetic.
[0448] [00448] A mixture of tert-butyl 1- (4- (2-hydroxyethyl) cyclohexyl) propane-1,3-diildicarbamate (1.95 mmol), ruthenium (III) chloride (0.008 g, 0.039 mmol), sodium periodate (1.67 g, 2.8 mmol) in carbon tetrachloride (10 ml), acetonitrile (10 ml) and water (10 ml) was stirred at RT for 2 h. The reaction mixture was cooled to 0 ° C and 0.5N HCl (10 ml) added, extracted with DCM, dried over Na2SO4. Purification by flashing column chromatography (eluent: 40% EtOAc in hexanes) gave the title acid as a white foam (0.67 g, 83%). LC / MC: 437.1 (MNa) +.
[0449] [00449] Synthesis of (R) -3- (2- (trans-4- (1,3-diaminopropyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [ and] [1,2] oxaborinin-8-carboxylic.
[0450] [00450] To a cooled (-25 ° C) solution of [(1S) -2- (3-tert-butoxycarbonyl-2-methoxy-phenyl) -1-chloro-ethyl] boronic acid (+) pinanediolate (Example 48, Step 1, 0.41 g, 0.92 mmol) in THF (2 ml) LHMDS (1 ml, 1M in THF) was added by dropping under N2. Upon completion of the addition, the reaction was stirred at RT for 1.5 h. Meanwhile, in a separate vial, to a mixture of 2- (trans-4- (2,2,12,12-tetramethyl-4,10-dioxo-3,11-doxa-5,9-diazatridecan-6) acid -yl) cyclohexyl) acetic (0.38 g, 0.92 mmol) and HATU (0.38 g, 1 mmol) DMA (2 mL) and 4-methylmorpholine (0.11 mL) were added and the mixture The resulting mixture was stirred at RT under N2 for 1.5 h. After 1.5 h, the two solutions were mixed and stirred at RT overnight. After aqueous preparation, the residue was purified by FC chromatography (eluent: 30% EtOAc in 40%, 50% hexanes) to provide the product (0.25 g, 33%). LC / MS: 848.2 (MNa) +.
[0451] [00451] To a solution of the above product (0.22 g, 0.266 mmol) in 1,4-dioxane (0.6 ml) was added aqueous 3N HCl (3 ml). The resulting mixture was heated to 100 ° C for 3 h. After cooling to RT, the residue was extracted with ether and the aqueous residue was concentrated. Reverse phase HPLC and lyophilization of the collection provided the title compound as a white solid. LC / MS: 404.1 (MH) +. EXAMPLE 77: (R) -3- (2- (trans-4- (1,2-diaminoethyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carboxylic.
[0452] [00452] Synthesis of 2- (trans-4- (2,2,11,11-tetramethyl-4,9-dioxo-3,10-dioxa-5,8-diazadodecan-6-yl) cyclohexyl) acid acetic. Step 1: Synthesis of ethyl 2- (4-oxocyclohexyl) acetate.
[0453] [00453] To a solution of 2- (1,4-dioxaspospiro [4,5] decan-8-yl) ethyl acetate (from step 1, Example 77, 4.71 g, 19.44 mmol) in acetonitrile ( 45 mL) 6 N aqueous HCl solution (45 mL) was added. The resulting mixture was stirred at RT for 2 h and neutralized with solid NaHCO3 to pH 8, extracted with EtOAc. The organic phase was washed with brine, dried over Na2SO4. Filtration and evaporation to dryness provided the ketone product as a colorless oil (2.97 g, 78.1%). Step 2: Synthesis of ethyl 2- (4-methylenocyclohexyl) acetate.
[0454] [00454] To a cooled (0 ° C) suspension of methyl triphenylphosphonium bromide (8.58 g, 23.5 mmol) in THF (60 mL) was added COBu (3.17 g, 28.3 mmol) in portions under N2. The reaction was slowly warmed to RT and stirred for 1 h. The resulting mixture was cooled to 0 ° C and a solution of ethyl 2- (4-oxocyclohexyl) acetate (2.9 g, 15.7 mmol) in THF (15 ml) was added. The resulting mixture was stirred at RT for 2 h and at 50 ° C overnight. After cooling to RT, the reaction was quenched by the addition of saturated NH4Cl, extracted with EtOAc, washed with brine, dried over Na2SO4. Purification by flashing column chromatography (eluent: 20% EtOAc in hexanes) provided the product as a colorless oil (2.11 g, 73.7%). Step 3: Synthesis of ethyl 2- (trans-4- (hydroxymethyl) cyclohexyl) acetate.
[0455] [00455] 9-BBN (0.5 N in THF, 57.5 mL, 28.75 mmol) was added to a solution of 2- (4-methylene-cyclohexyl) ethyl acetate (2.10 g, 11, 5 mmol) in THF (20 mL) at 0 ° C under N2. The mixture was heated to RT and stirred at RT for 3 h. The reaction mixture was cooled to 0 ° C and a mixture of 20% NaOAc solution (40 ml) and 30% H2O2 (30 ml) was added by dropping. The resulting mixture was heated to RT and stirred for 40 min, cooled quickly with saturated NH4Cl solution, diluted with EtOAc, and separated. The organic phase was washed with saturated Na2S2O3 solution, brine and dried over Na2SO4. Purification by sparkling column chromatography (eluent: 30% EtOAc in 40% hexanes) gave the product alcohol as a colorless oil (1.66 g, 72.1%). LC / MC: 201.1 (MH) +. Step 4: Synthesis of ethyl 2- (trans-4-formylcyclohexyl) acetate.
[0456] [00456] A solution of DMSO (0.64 mL, 9.06 mmol) in DCM (2 mL) was added by dripping to a pre-cooled (-78 ° C) solution of oxalyl chloride (0.85 mL, 9.9 mmol) in DCM (2 mL) under N2. After stirring at -78 ° C for 10 min, a solution of ethyl 2- (trans-4- (hydroxymethyl) cyclohexyl) acetate (1.65 g, 8.23 mmol) in DCM (12 mL) was added by drip. The resulting solution was stirred at -78 ° C for 15 min, and TEA (6.89 ml) was added. The solution was stirred at -78 ° C for 15 min and at RT for 20 min. The reaction was diluted with DCM, washed with 1M HCl solution and brine, dried over Na2SO4. Purification by flashing column chromatography (eluent: 20% EtOAc in 30% hexanes) provided the ketone product as yellow oil (0.76 g, 47%). LC / MC: 199.1 (MH) +. Step 5: Synthesis of ethyl 2- (trans-4- (amino (cyano) methyl) cyclohexyl) acetate.
[0457] [00457] A mixture of 2- (trans-4-formylcyclohexyl) ethyl acetate (0.75 g, 3.78 mmol), NaCN (0.21 g, 4.28 mmol), saturated aqueous NH4OH solution (0.53 ml) and NH4Cl (0.24 g) in EtOH (22 ml) and water (11 ml) was heated to 70 ° C overnight. After removing the organic solvent, the residue was extracted with EtOAc, washed with saturated NaHCO3 solution and brine, dried over Na2SO4. Filtration and evaporation to dryness provided the crude product. LC / MC: 225.1 (MH) +. Step 6: Synthesis of 2- (trans-4- (tert-butoxycarbonylamino) (cyano) methyl) cyclohexyl) ethyl acetate.
[0458] [00458] To a solution of 2- (trans-4- (amino (cyano) methyl) cyclohexyl) ethyl acetate (3.70 mmol) in THF (25 mL) was added di-tert-butyldicarbonate (1, 25 g, 5.55 mmol) and NaHCO3 (0.62 g, 7.4 mmol). The resulting mixture was stirred at RT overnight. Aqueous preparation and purification by sparkling column chromatography (eluent: 30% EtOAc in hexanes) provided the product (0.54 g, 45%). LC / MC: 347.1 (MNa) +. Step 7: Synthesis of ethyl 2- (trans-4- (2-amino-1- (tert-butoxycarbonylamino) ethyl) cyclohexyl) acetate
[0459] [00459] A solution of 2- (trans-4- (tert-butoxycarbonylamino) (cia-no) methyl) cyclohexyl) ethyl acetate (1.63 mmol) in acetic acid (15 mL) was added Pd (OH ) 2 (0.2 g). The resulting mixture was hydrogenated at RT under 55 psi for 3 days. Filtration and evaporation to dryness provided the crude product. LC / MC: 329.1 (MH) +. Step 8: Synthesis of 2- (trans-4- (2,2,11,11-tetramethyl-4,9-dioxo-3,10-dioxa-5,8-diazadodecan-6-yl) cyclohexyl) acetate of ethyl.
[0460] [00460] A mixture of ethyl 2- (trans-4- (2-amino-1- (tert-butoxycarbonyl-mino) ethyl) cyclohexyl) acetate (1.63 mmol), di-tert-butyldicar-bonate (0.44 g, 1.96 mmol) and TEA (2.27 mL, 16.3 mmol) in DCM (15 mL) was stirred at RT overnight. The reaction was quenched by the addition of saturated NH4G solution and separated. The organic phase was washed with 0.5N HCl, brine, and dried over Na2SO4. Purification by sparkling column chromatography (eluent: 20% EtOAc in 30% hexanes) provided the product as a colorless oil (0.58 g, 83%). LC / MC: 451.1 (MNa) +. Step 9: Synthesis of 2- (trans-4- (2,2,11,11-tetramethyl-4,9-dioxo- 3.10-dioxa-5,8-diazadodecan-6-yl) acetic acid.
[0461] [00461] To a solution of 2- (trans-4- (2,2,11,11-tetramethyl-4,9-dioxo- 3.10-dioxa-5,8-diazadodecan-6-yl) cyclohexyl) acetate of ethyl (0.58 g, 1.35 mmol) in THF (5 mL) and MeOH (5 mL) was added 3N NaOH (2.25 mL, 6.75 mmol). The resulting mixture was stirred at RT for 3 h. After removing the volatiles through evaporation, the residue was acidified by 0.5 N HCl solution to pH ~ 4.5 and extracted with EtOAc, dried over Na2SO4. Filtration and evaporation to dryness provided the acid in a quantitative yield. LC / MS: 423.1 (MNa) +.
[0462] [00462] Synthesis of (R) -3- (2- (trans-4- (1,2-diaminoethyl) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [ and] [1,2] oxaborinin-8-carboxylic.
[0463] [00463] Prepared from 2- (trans-4- (2,2,11,11-tetramethyl-4,9-dioxo-3,10-dioxa-5,8-diazadodecan-6-yl) hexyl) acetic after the coupling and deprotection procedure described in Example 76. The product was obtained as a white solid. LC / MS: 390.1 (MH) +. EXAMPLE 78: (R) -3- (2- (trans-4- (2- (ethylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e ] [1,2] oxaborinin-8-carboxylic.
[0464] [00464] Synthesis of (R) -3- (2- (trans-4- (2- (ethylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborin-8-carboxylic.
[0465] [00465] Prepared from (R) -3- (2- (trans-4- (2-aminoethyl-mino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H- benzo [e] [1,2] oxaborinin-8-carboxylic and acetaldehyde after the procedure described in Example 71. The product was purified using reverse phase HPLC to provide the title compound. ESI-MS m / z 418 (MH) +. EXAMPLE 79: (R) -3- (2- (trans-4- (2- (dimethylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo [e ] [1,2] oxaborinin-8-carboxylic. Step 1: Synthesis of 2- (trans-4- (tert-butoxycarbonyl (2- (dimethyl-mino) ethyl) amino) cyclohexyl) acetic acid.
[0466] [00466] To 2- (trans-4- (tert-butoxycarbonylamino) cyclohexyl) acetic acid (1.47 g) in DMF (10 ml) was added Na2CO3 (0.907 g) and benzyl bromide (0.75 ml) ). The resulting reaction mixture was stirred at RT overnight. Water was then added and extracted with EtOAc. The organic phase was dried and concentrated to provide the desired product as a white solid (1.60 g). To this solid was added 4N HCl (10 ml) and the reaction mixture was stirred at RT for 1 h. Diethyl ether was then added to the reaction mixture to precipitate the 2- (trans-4-aminocyclohexyl) benzyl acetate HCl salt as a white solid.
[0467] [00467] To the salt of 2- (trans-4-aminocyclohexyl) benzyl acetate HCl (860 mg) in DMF was added K2CO3 (414 mg) and salt of 2-bromo-N, N-dimethylethanamine HBr (700 mg ). The resulting reaction mixture was stirred at 60 ° C overnight. Water was then added and extracted with EtOAc. The organic phase was dried and concentrated to provide the crude product which was used directly in the next step.
[0468] [00468] To the above product in DCM (20 ml) was added TEA (1 ml) and di-tert-butyl dicarbonate (1.5 g). The reaction mixture was stirred at RT overnight. The organic phase was washed with brine, dried and concentrated. The residue was purified by HPLC. To this product in MeOH (10 ml) was added Pd / C (10%, 50 mg) and the reaction mixture was stirred under a hydrogen atmosphere overnight. The catalyst was filtered through Celite buffer and the solvent removed under reduced pressure to provide 2- (trans-4- (tert-butoxycarbonyl) (2- (dimethylamino) ethyl) amino) cyclohexyl) acetic acid as a yellow foam (250 mg). Step 2: Synthesis of 3 - ((2R) -2- (2- (trans-4- (tert-butoxycarbonyl (2- (dimethylamino) ethyl) amino) cyclohexyl) acetamido) -2- (2, 9,9-trimethyl-3,5-dioxa-4-bora-tricycle [6,1,1,02,6] dec-4-yl) ethyl) tert-butyl -2-methoxybenzoate
[0469] [00469] Prepared from 2-methoxy-3- (2,9,9-trimethyl-3,5-dioxa-4-bora-tricycle) tert-butyl ester [6,1,1,02'6] dec-4-ylmethyl) -benzoic acid and 2- (trans-4- (tert-butoxycarbonyl (2- (dimethylamino) ethyl) amino) cyclohexyl) acetic acid after the procedure described in Step 1 of Example 1. Step 3: Synthesis of (R) -3- (2- (trans-4- (2- (dimethylamino) ethylamino) cyclohexyl) acetamido) -2-hydroxy-3,4-dihydro-2H-benzo acid [e] [1,2] oxaborinin-8-carboxylic.
[0470] [00470] To the compound of step 2 (40 mg) was added 3N HCl (2 ml) and the resulting reaction mixture was heated to reflux for 1 h. The solvents were then removed in vacuo and the residue purified by preparative reverse phase HPLC and dried using lyophilization. ESI-MS m / z 418 (MH) +. Table 1. Examples of compounds
[0471] [00471] To prepare a parenteral pharmaceutical composition suitable for administration by injection, 100 mg of a compound of Formula I or Formula Ia, or a pharmaceutically acceptable salt soluble in water thereof, is dissolved in DMSO and then mixed with 10 ml of saline sterile 0.9%. The mixture is incorporated in a dosage unit suitable for administration by injection. EXAMPLE 108: Oral Composition of a Formula I or Formula Ia Compound
[0472] [00472] To prepare a pharmaceutical composition for oral delivery, 400 mg of a compound of Formula I or Formula Ia and the following ingredients are intimately mixed and pressed into individual marked tablets. Tablets Formulation
[0473] [00473] The following ingredients are mixed intimately and loaded into a solid structure gelatin capsule. Capsule Formulation
[0474] [00474] For the β-lactamases SHV-5, Kpc-2, p99AmpC and OXA-1, the bacterial cells that carry the expression plasmids (expressed as unlabeled native proteins) for the individual β-lactamases were cultured in 1 L Superbroth (Teknova Inc. Hollister, CA) supplemented with 100 μg / mL kanamycin selection and 1x 5052 (0.5% glycerol, 0.05% glucose and 0.2% α-lactose) at 35 ° C for 18 to 20 hours. The cells were harvested by centrifugation (4000 x g, 4 ° C, 20 min), resuspended in 50 ml of 10 mM HEPES pH 7.5 (1/20 of the initial volume). The cells were subjected to sonication lysis (5 pulses of 45 seconds) at 45 W on ice. Lysates were clarified by centrifugation at 10,000 x g for 40 minutes at 4 ° C. The samples were diluted 5 times in 50 mM sodium acetate, pH 5.0, stored overnight at 4 ° C, after which they were centrifuged at 10,000 xg for 30 minutes to clarify, and filtered through 0.45 filters. μm. The samples were loaded onto a 5 mL Capto S Sepharose cation exchange column (GE Healthcare) pre-equilibrated with 50 mM sodium acetate pH 5.0. The column was washed with 5 column volumes of 50 mM sodium acetate pH 5.0 to wash the unbound protein and a linear gradient of NaCl (0 to 500 mM) was used to elute the protein (more than 16 CV) at from the column. Fractions were tested for β-lactamase activity using Centa (Calbiochem, Gibbstown, NJ) or Nitrocefin (EMD Millipore chemicals, Darmstadt, Germany) as a β-lactamase reporter substrate for activity in isolated fractions. The active fractions were combined, concentrated and further purified by gel filtration chromatography on a Superdex 75 prep grade gel filtration column (GE Healthcare, Piscataway, NJ) pre-equilibrated with 50 mM Hepes pH 7.5, 150 mM NaCl. The active fractions were combined concentrated, quantified by BCA protein determination (Thermo Scientific, Rockford, IL), subjected to dialysis in PBS and frozen at -80 ° C in 20% glycerol until use.
[0475] [00475] For Vim 2-metallo β-lactamase, the procedure was identical with the following exceptions, firstly the protein was not adjusted with respect to pH to pH 5 with 50 mM sodium acetate, secondly, to The chromatography step was changed to a 5 mL Q-Sepharose anion exchange column pre-equilibrated with 50 mM Hepes pH 7.5, and protein elution was achieved by a linear gradient of NaCl (0 to 600 mM). Finally, VIM-2 purification required a second operation (3rd step) on the Q Sepharose anion exchange column to achieve an acceptable purity (> 90%). Inhibition of β-Lactamase.
[0476] [00476] To determine the level of inhibition of β-lactamase enzymes, the compounds were diluted with PBS at pH 7.4, to produce concentrations ranging from 100 to 0.00005 μΜ in 96 well microtiter plates. An equal volume of diluted enzyme feedstock was added, and the plates were incubated at 37 ° C for 15 min. Nitrocefine was used as a substrate for p99 AmpC, VIM-2 and OXA-1 and dispensed in each reservoir at a final concentration of 100 μM. The absorbance at 486 nm was immediately monitored for 10 min using a Biotek Powerwave XS2 microplate spectrophotometer using the GEN5 softweare package (Biotek Instruments, Winooski VT). Similarly, imipenem was used as a substrate for KPC-2 and Cefotaxime was used for SHV-5, while changes in absorbance after hydrolysis of the β-lactam ring were monitored at 300 nm and 260 nm, respectively, in the microtiter assay plates of 96 UV-transparent reservoirs. The maximum rates of hydrolysis were compared to those in the control reservoirs (without inhibitor), and the percent inhibition of the enzyme was calculated for each concentration of inhibitor. The inhibitor concentration necessary to reduce the initial rate of substrate hydrolysis by 50% (IC50) was calculated as the residual activity of β-lactamase at 486 nm, using the GraFit version 7 kinetics software package (Erithacus Software, Surrey, UK ). EXAMPLE II: Inhibition of Various β-Lactamases through Exemplary Compounds
[0477] [00477] Using the methodology described above, the examples of the present invention were evaluated for their ability to inhibit β-lactamase enzymes from all four Ambler classifications (A through D). The results of these assays are summarized in Table 3 for the representative enzymes across different subtypes (note that SHV-5 represents an Ambler Class A Extended Spectrum β-Lactamases, KPC-2 exemplifies a carbapenemase Class A, P99 represents cAMP Class C chromosome, OXA-1 represents a Class D oxacillinase and VIM-2 represents a class B zinc-dependent metallo-lactamase that also has carbapenemase activity), where A represents an IC50 of 10 to 100 μΜ, B represents an IC50 of 1 to 10 μΜ, C represents an IC 50 of 0.1 to 1 μΜ, and D represents an IC 50 of <0.1 μΜ. NT = Not tested. Table 3. Inhibition of Various β-Lactamases through Compounds
[0478] [00478] To determine the ability of test compounds to potentiate the growth inhibition of bacterial strains that produce beta-lactamase enzymes, broth microdilution assays based on classic MIC cells were employed. Six bacterial strains that produce beta-lactamase enzymes were used: E. coli which expresses Class A Extended Spectrum Beta-Lactamase (ESBL) CTX-M-15, E. cloacae which expresses Class C P99, K. pneumoniae which expresses class A carbapenemase KPC-2, P. aeruginosa expressing class B carbapenemase VIM-2, K. pneumoniae expressing class A carbapenemase KPC-2 and class B carbapenemase VIM-4, and S. aureus which produces Class A PC-1 penicillinase. The test was conducted in Cation Adjusted Mueller Hinton Broth (CAMHB, BD # 212322, BD Diagnostic Systems, Sparks, MD). The strains of bacteria were grown for 3 to 5 hours in CAMBH broth. The test compounds were added to a microtiter plate in 2-fold serial dilutions with CAMHB in a final concentration range of 32 μg / mL to 0.25 μg / mL. A CAMHB overlay containing a beta-lactam was added to the compounds at a final static concentration of 4 μg / mL. Ceftazidime (CAZ, Sigma # C3809-1G, Sigma-Aldrich, St. Louis, MO) was used as the partner antibiotic for E. coli that expresses Ambler Class A ESBL CTX-M-15 (MIC alone> 128 μg / mL) , and E. cloacae which expresses Class C P99 (MIC alone = 128 μg / mL). Meropenem (Mero, USP # 1392454, US Pharmacopeia, Rockville, MD) was used as the partner antibiotic for K. pneumoniae that expresses Ambler Class A carbapenemase KPC-3 (MIC alone> 128 μg / mL), P. aeruginosa that expresses Class Carbapenemase VIM-2 (MIC alone = 16 μg / mL), and K. pneumoniae expressing Ambler Class A carbapenemase KPC-2 and Ambler Class B carbapenemase VIM-4 (MIC alone = 64 μg / mL). Piperacillin (Pip, Fisher # ICN15626801, MP Biomidicals, Solon, OH) was used as the partner antibiotic for S. aureus that produces Class A penicillinase PC-1 (MIC alone = 64 μg / mL). The titration of test compounds with MIC reading indicates the concentration of the test article necessary to sufficiently inhibit the activity of the beta-lactamase enzyme and protect the intrinsic antibacterial activity of the beta-lactam. In addition to the titration of the test compounds, the MICs of a control panel of beta-lactams are also tested to ensure that the strains behave consistently from test to test. As soon as the test compound and antibiotics are added, the plates can be inoculated according to the CLSI broth microdilution method. After inoculation, the plates are incubated for 16 to 20 hours at 37 ° C, then the Minimum Inhibitory Concentration (MIC) of the test compound is determined visually.
[0479] [00479] Using the methodology described above, the examples of the present invention were evaluated for their ability to inhibit the growth of β-lactamase producing bacteria in the presence of a β-lactam antibiotic.
[0480] [00480] Representative results are shown in Table 3 where A represents a MIC> 16 μg / mL, B represents a MIC between 1 and 16 μg / mL inclusive, and C represents a MIC value <1 μg / mL. NT = Not tested. EXAMPLE IV: In Vitro Antibacterial Activity of Exemplary Compounds
[0481] [00481] Using the methodology described above in Example III, exemplary Formula I or Formula Ia compounds were evaluated for their ability to inhibit the growth of β-lactamase-producing bacteria in the presence of a β-lactam antibiotic.
[0482] [00482] Representative results are shown in Table 4 where A represents a MIC of the β-lactam antibiotic fixed in the presence of> 32 μg / mL of an β-lactamase inhibitor of the exemplary compounds, B represents the MIC in the presence of 8 and 32 μg / mL of an β-lactamase inhibitor of exemplary compounds, and C represents MIC in the presence of <4 μg / mL of an β-lactamase inhibitor of exemplary compounds. NT = Not tested. Table 4: Broad-spectrum inhibition of bacterial growth. The MIC of the example compounds of the invention in the presence of a fixed amount (4 μg / ml) of the β-lactam antibiotics called ceftazidime (CAZ), meropenem (Mero), Piperacillin (Pip).
[0483] [00483] Although the preferred embodiments of the present invention have been presented and described herein, it will be obvious to those skilled in the art that such modalities are provided by way of example only. Numerous variations, changes and substitutions will now occur for those skilled in the art without departing from the invention. It should be understood that, several alternatives to the modalities of the invention described herein can be employed in the practice of the invention. It is intended that the following claims define the scope of the invention and that the methods and structures within the scope of these claims and their equivalents are thus covered.

权利要求:
Claims (12)
[0001]
Compound, characterized by the fact that it has Formula (I) or Formula (Ia), a pharmaceutically acceptable salt, stereoisomer or N-oxide of this:
[0002]
Compound according to claim 1, characterized by the fact that Ra, Rb and Rc are independently hydrogen, fluorine or chlorine.
[0003]
Compound according to claim 1 or 2, characterized by the fact that R3 is hydrogen.
[0004]
Compound according to any one of claims 1 to 3, characterized in that X1 and X2 are -OH.
[0005]
Compound according to any one of claims 1 to 4, characterized in that Rd is hydrogen or C1-C4 alkyl.
[0006]
Compound according to any one of claims 1 to 5, characterized by the fact that: L is -CR1R2-; M is a bond; and men are 0.
[0007]
Compound according to any one of claims 1 to 6, characterized by the fact that:
[0008]
Compound, according to claim 1, characterized by the fact that it is selected from the group represented by the following structures:
[0009]
Compound according to claim 1, characterized by the fact that it is:
[0010]
Pharmaceutical composition, characterized in that it comprises a compound, as defined in any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer, N-oxide or isomer thereof, and a pharmaceutically acceptable excipient.
[0011]
Pharmaceutical composition according to claim 10, characterized by the fact that it further comprises a beta-lactam antibiotic.
[0012]
Use, of a compound, as defined in any one of claims 1 to 9, or of a pharmaceutical composition, as defined in claim 10 or 11, optionally in combination with a beta-lactam antibiotic, characterized in that it is for the preparation of a medicament for treating a bacterial infection in a mammalian individual.

类似技术:

---

公开号 | 公开日 | 专利标题

---

ES2878118T3|2021-11-18|Beta-lactamase inhibitors

---

US10125152B2|2018-11-13|Beta-lactamase inhibitors

---

EP2970340B1|2020-02-12|Beta-lactamase inhibitors

同族专利:

---

公开号 | 公开日

---

AU2013355110B2|2017-11-09|

---

JP2018184409A|2018-11-22|

---

US10669290B2|2020-06-02|

---

RU2654692C2|2018-05-22|

---

US8912169B2|2014-12-16|

---

US20180194783A1|2018-07-12|

---

AU2013355110A1|2015-07-02|

---

CA2893943C|2021-03-02|

---

US9422314B2|2016-08-23|

---

CA2893943A1|2014-06-12|

---

US10214547B2|2019-02-26|

---

IL239212A|2019-10-31|

---

KR102147420B1|2020-08-25|

---

EP3922635A1|2021-12-15|

---

JP6403219B2|2018-10-10|

---

US20160326189A1|2016-11-10|

---

US20140171390A1|2014-06-19|

---

HK1217334A1|2017-01-06|

---

ES2878118T3|2021-11-18|

---

PL2928898T3|2021-10-25|

---

CN105026407A|2015-11-04|

---

KR20150109348A|2015-10-01|

---

RU2015126912A|2017-01-12|

---

ZA201504200B|2017-09-27|

---

WO2014089365A1|2014-06-12|

---

US20190225628A1|2019-07-25|

---

EP2928898A4|2016-06-29|

---

PT2928898T|2021-07-05|

---

EP3922635A4|2021-12-15|

---

EP2928898B1|2021-04-14|

---

CN105026407B|2017-09-08|

---

JP2016502973A|2016-02-01|

---

US20150291630A1|2015-10-15|

---

EP2928898A1|2015-10-14|

---

BR112015013123A2|2017-07-11|

---

IL239212D0|2015-07-30|

---

HRP20210879T1|2021-08-20|

---

US9828391B2|2017-11-28|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

NZ177159A|1974-04-20|1978-03-06|Beecham Group Ltd|Clavulanic acid, salts, esters and preparation thereof from streptomyces clavuligerus: pharmaceutical compositions|

---

IT1283467B1|1996-07-19|1998-04-21|Menarini Farma Ind|1,2 CYCLOALKANE DERIVATIVES REPLACED AS INHIBITORS OF THE THROMBIN, PROCEDURE FOR THEIR PREPARATION AND THEIR USE IN FORMULATIONS|

---

DE60004685T2|1999-09-17|2004-07-29|Abbott Gmbh & Co. Kg|PYRAZOLOPYRIMIDINE AS A MEDICINAL PRODUCT|

---

US7271186B1|2002-12-09|2007-09-18|Northwestern University|Nanomolar β-lactamase inhibitors|

---

AU2004256841A1|2003-06-10|2005-01-20|Fulcrum Pharmaceuticals, Inc.|Beta-lactamase inhibitors and methods of use thereof|

---

ES2457593T3|2004-03-30|2014-04-28|Millennium Pharmaceuticals, Inc.|Bortezomib synthesis|

---

CN1965838A|2005-11-17|2007-05-23|李海超|Pharmaceutical composition containing cefpiramide and beta-lactamase inhibitor|

---

CA2629570C|2005-12-07|2012-11-27|Basilea Pharmaceutica Ag|Useful combinations of monobactam antibiotics with beta-lactamase inhibitors|

---

CN101129382B|2006-08-25|2013-12-25|天津和美生物技术有限公司|Antibiotic compound containing beta-lactam antibiotic and buffering component|

---

US20100120715A1|2007-11-13|2010-05-13|Burns Christopher J|Beta-lactamase inhibitors|

---

TW200930707A|2007-11-13|2009-07-16|Protez Pharmaceuticals Inc|Beta-lactamase inhibitors|

---

RS53862B1|2008-01-18|2015-08-31|Merck Sharp & Dohme Corp.|Beta-lactamase inhibitors|

---

AR076667A1|2009-05-12|2011-06-29|Novartis Int Pharm Ltd|BETA-LACTAMASA INHIBITORS|

---

KR20130031233A|2009-07-28|2013-03-28|아나코르 파마슈티칼스 인코포레이티드|Trisubstituted boron-containing molecules|

---

MX348653B|2010-08-10|2017-06-21|Rempex Pharmaceuticals Inc |Cyclic boronic acid ester derivatives and therapeutic uses thereof.|

---

EP2768503A1|2011-07-26|2014-08-27|Wockhardt Limited|Pharmaceutical compositions comprising beta-lactam antibiotic, sulbactam and beta-lactamase inhibitor|

---

SG11201400293UA|2011-09-04|2014-08-28|Glytech Inc|Glycosylated polypeptide and drug composition containing said polypeptide|

---

WO2013053372A1|2011-10-13|2013-04-18|Therabor Pharmaceuticals|Boronic acid inhibitors of beta-lactamases|

---

KR101288157B1|2011-12-07|2013-07-19|백병하|A Combined antibiotics comprising cepha antibiotics and beta-lactamase inhibitor|

---

SG11201403254QA|2011-12-22|2014-07-30|Ares Trading Sa|Alpha-amino boronic acid derivatives, selective immunoproteasome inhibitors|

---

EP2814483A2|2012-02-15|2014-12-24|Rempex Pharmaceuticals, Inc.|Methods of treating bacterial infections|

---

BR112015012909A2|2012-12-03|2017-07-11|Hoffmann La Roche|substituted triazole boronic acid compounds|

---

AU2013355110B2|2012-12-07|2017-11-09|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

US9101638B2|2013-01-04|2015-08-11|Rempex Pharmaceuticals, Inc.|Boronic acid derivatives and therapeutic uses thereof|

---

US9642869B2|2013-01-04|2017-05-09|Rempex Pharmaceuticals, Inc.|Boronic acid derivatives and therapeutic uses thereof|

---

MX2015008627A|2013-01-04|2015-09-23|Rempex Pharmaceuticals Inc|Boronic acid derivatives and therapeutic uses thereof.|

---

WO2014110442A1|2013-01-10|2014-07-17|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

WO2014151958A1|2013-03-14|2014-09-25|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

US9120796B2|2013-10-02|2015-09-01|Cubist Pharmaceuticals, Inc.|B-lactamase inhibitor picoline salt|

---

WO2015157618A1|2014-04-11|2015-10-15|The Texas A&M University System|Novel inhibitors of the new delhi metallo beta lactamase |

---

EP3145936B1|2014-05-19|2020-09-30|Qpex Biopharma, Inc.|Boronic acid derivatives and therapeutic uses thereof|

---

US20150361107A1|2014-06-11|2015-12-17|VenatoRx Pharmaceuticals, Inc.|Orally bioavailable beta-lactamase inhibitors|

---

US20150361108A1|2014-06-11|2015-12-17|VenatoRx Pharmaceuticals, Inc.|Orally bioavailable beta-lactamase inhibitors|

---

US9511142B2|2014-06-11|2016-12-06|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

RU2686740C2|2014-06-11|2019-04-30|Венаторкс Фармасьютикалс, Инк.|Beta-lactamase inhibitors|

---

US9949982B2|2014-09-04|2018-04-24|Shionogi & Co., Ltd.|Preparation containing cephalosporin having a catechol moiety|

---

WO2016100043A1|2014-12-19|2016-06-23|Rempex Pharmaceuticals, Inc.|Apparatus and continuous flow process for production of boronic acid derivatives|

---

CN108472284A|2015-09-11|2018-08-31|维纳拓尔斯制药公司|Beta-lactamase inhibitor|

---

WO2017100537A1|2015-12-10|2017-06-15|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

CN105801610B|2016-04-19|2018-05-18|武汉维舜医药科技有限公司|New-type wide-spectrum beta-lactamase inhibitor|

---

US10889600B2|2016-08-04|2021-01-12|VenatoRx Pharmaceuticals, Inc.|Boron-containing compounds|

---

MA47743A|2017-03-06|2020-01-15|Venatorx Pharmaceuticals Inc|SOLID FORMS AND COMBINATION COMPOSITIONS INCLUDING A BETA-LACTAMASE INHIBITOR AND THEIR USES|

---

US20200157123A1|2017-05-26|2020-05-21|VenatoRx Pharmaceuticals, Inc.|Penicillin-binding protein inhibitors|

---

WO2018218154A1|2017-05-26|2018-11-29|VenatoRx Pharmaceuticals, Inc.|Penicillin-binding protein inhibitors|

---

US11247965B2|2017-12-11|2022-02-15|VenatoRx Pharmaceuticals, Inc.|Hepatitis B capsid assembly modulators|

---

EP3802551A4|2018-05-25|2022-03-02|Venatorx Pharmaceuticals Inc|Penicillin-binding protein inhibitors|MX348653B|2010-08-10|2017-06-21|Rempex Pharmaceuticals Inc |Cyclic boronic acid ester derivatives and therapeutic uses thereof.|

---

US9012491B2|2011-08-31|2015-04-21|Rempex Pharmaceuticals, Inc.|Heterocyclic boronic acid ester derivatives and therapeutic uses thereof|

---

US10561675B2|2012-06-06|2020-02-18|Rempex Pharmaceuticals, Inc.|Cyclic boronic acid ester derivatives and therapeutic uses thereof|

---

AU2013355110B2|2012-12-07|2017-11-09|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

MX2015008627A|2013-01-04|2015-09-23|Rempex Pharmaceuticals Inc|Boronic acid derivatives and therapeutic uses thereof.|

---

US9241947B2|2013-01-04|2016-01-26|Rempex Pharmaceuticals, Inc.|Boronic acid derivatives and therapeutic uses thereof|

---

US9642869B2|2013-01-04|2017-05-09|Rempex Pharmaceuticals, Inc.|Boronic acid derivatives and therapeutic uses thereof|

---

US9101638B2|2013-01-04|2015-08-11|Rempex Pharmaceuticals, Inc.|Boronic acid derivatives and therapeutic uses thereof|

---

WO2014110442A1|2013-01-10|2014-07-17|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

WO2014151958A1|2013-03-14|2014-09-25|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

WO2015171398A1|2014-05-05|2015-11-12|Rempex Pharmaceuticals, Inc.|Salts and polymorphs of cyclic boronic acid ester derivatives and therapeutic uses thereof|

---

ES2750805T3|2014-05-05|2020-03-27|Rempex Pharmaceuticals Inc|Synthesis of boronate salts and uses thereof|

---

EP3145936B1|2014-05-19|2020-09-30|Qpex Biopharma, Inc.|Boronic acid derivatives and therapeutic uses thereof|

---

RU2686740C2|2014-06-11|2019-04-30|Венаторкс Фармасьютикалс, Инк.|Beta-lactamase inhibitors|

---

US9511142B2|2014-06-11|2016-12-06|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

US20150361108A1|2014-06-11|2015-12-17|VenatoRx Pharmaceuticals, Inc.|Orally bioavailable beta-lactamase inhibitors|

---

AU2015284307A1|2014-07-01|2017-02-02|Rempex Pharmaceuticals, Inc.|Boronic acid derivatives and therapeutic uses thereof|

---

WO2016081297A1|2014-11-18|2016-05-26|Rempex Pharmaceuticals, Inc.|Cyclic boronic acid ester derivatives and therapeutic uses thereof|

---

WO2016100043A1|2014-12-19|2016-06-23|Rempex Pharmaceuticals, Inc.|Apparatus and continuous flow process for production of boronic acid derivatives|

---

CN107207472B|2014-12-23|2020-08-14|卑尔根生物股份公司|Inhibitors of AKT kinase|

---

WO2016149393A1|2015-03-17|2016-09-22|Rempex Pharmaceuticals, Inc.|Boronic acid derivatives and therapeutic uses thereof|

---

CN108472284A|2015-09-11|2018-08-31|维纳拓尔斯制药公司|Beta-lactamase inhibitor|

---

WO2017100537A1|2015-12-10|2017-06-15|VenatoRx Pharmaceuticals, Inc.|Beta-lactamase inhibitors|

---

CN105801610B|2016-04-19|2018-05-18|武汉维舜医药科技有限公司|New-type wide-spectrum beta-lactamase inhibitor|

---

DK3478693T3|2016-06-30|2021-09-20|Qpex Biopharma Inc|BORONIC ACID DERIVATIVES AND THERAPEUTIC USES THEREOF|

---

US10889600B2|2016-08-04|2021-01-12|VenatoRx Pharmaceuticals, Inc.|Boron-containing compounds|

---

MA47743A|2017-03-06|2020-01-15|Venatorx Pharmaceuticals Inc|SOLID FORMS AND COMBINATION COMPOSITIONS INCLUDING A BETA-LACTAMASE INHIBITOR AND THEIR USES|

---

TW201841642A|2017-04-28|2018-12-01|日商大日本住友製藥股份有限公司|Heterocyclic derivative|

---

WO2018218154A1|2017-05-26|2018-11-29|VenatoRx Pharmaceuticals, Inc.|Penicillin-binding protein inhibitors|

---

US20200157123A1|2017-05-26|2020-05-21|VenatoRx Pharmaceuticals, Inc.|Penicillin-binding protein inhibitors|

---

SG11202003762VA|2017-11-01|2020-05-28|Melinta Therapeutics Inc|Synthesis of boronate ester derivatives and uses thereof|

---

JPWO2019208797A1|2018-04-27|2021-05-27|大日本住友製薬株式会社|Oxo-substituted compound|

---

EP3802551A4|2018-05-25|2022-03-02|Venatorx Pharmaceuticals Inc|Penicillin-binding protein inhibitors|

---

EP3849562A1|2018-09-12|2021-07-21|Venatorx Pharmaceuticals, Inc.|Combination compositions comprising a beta-lactamase inhibitor|

---

BR112021019656A2|2019-04-02|2022-01-18|Venatorx Pharmaceuticals Inc|Solid forms of an orally dispensed beta-lactamase inhibitor and uses thereof|

---

US11058695B2|2019-11-08|2021-07-13|Myongji University Industry And Academia Cooperation Foundation|Inhibitor of carbapenem-hydrolyzing class D beta-lactamases|

---

WO2022037680A1|2020-08-20|2022-02-24|辉诺生物医药科技（杭州）有限公司|Boracic acid compound|

法律状态:
2018-03-06| B07D| Technical examination (opinion) related to article 229 of industrial property law [chapter 7.4 patent gazette]|

---

2018-11-21| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

---

2019-07-16| B07E| Notification of approval relating to section 229 industrial property law [chapter 7.5 patent gazette]|Free format text: NOTIFICACAO DE ANUENCIA RELACIONADA COM O ART 229 DA LPI |

---

2020-01-14| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

---

2020-05-26| B06A| Patent application procedure suspended [chapter 6.1 patent gazette]|

---

2020-12-15| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

---

2021-02-17| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 05/12/2013, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

---

申请号 | 申请日 | 专利标题

---

US201261734900P| true| 2012-12-07|2012-12-07|

---

US61/734,900|2012-12-07|

---

US201361783238P| true| 2013-03-14|2013-03-14|

---

US61/783,238|2013-03-14|

---

PCT/US2013/073428|WO2014089365A1|2012-12-07|2013-12-05|Beta-lactamase inhibitors|

---