P38 MAP kinase inhibitors

专利摘要:

公开号:ES2605727T9
申请号:ES07732629T
申请日:2007-05-01
公开日:2020-12-11
发明作者:David Charles Festus Moffat；Stéphane Pintat；Stephen Davies
申请人:Macrophage Pharma Ltd；
IPC主号:

专利说明:

[0002] P38 MAP kinase inhibitors
[0004] The present invention relates to a series of amino acid and amino acid ester compounds, to compositions containing them, to processes for their preparation and to their use in medicine as p38 MAP kinase inhibitors for the treatment of autoimmune and inflammatory diseases, including rheumatoid arthritis, psoriasis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, chronic obstructive pulmonary disease, asthma, multiple sclerosis, diabetes, atopic dermatitis, graft-versus-host disease, systemic lupus erythematosus, and others.
[0006] Background of the invention
[0008] Inappropriate activation of leukocytes, including monocytes, macrophages, and neutrophils that results in the production of high levels of cytokines, such as TNF-a, IL1-p, and IL-8, is a feature of the pathogenesis of several inflammatory diseases, including rheumatoid arthritis, ulcerative colitis, Crohn's disease, chronic obstructive pulmonary disease (COPD), asthma, and psoriasis. The production of cytokines by inflammatory cells is the result of the response to a variety of external stimuli, which give rise to the activation of a series of intracellular signaling mechanisms. Notable among these is the mitogen-activated protein kinase (MAPK) superfamily consisting of highly conserved signaling kinases that regulate cell growth, differentiation, and responses to stress. Mammalian cells contain at least three families of MAPK: the extracellular signal-regulated MAPK kinases (ERK) p42 / 44, the NH2-terminal c-Jun kinases (JNK), and the MAPK p38 (also called p38a / Mpk2 / RK / SAPK2a / CSBP1 / 2). P38 MAPK was first cloned after its identification as a tyrosine phosphorylated kinase after lipopolysaccharide (LPS) stimulation of monocytes [Han et al., Science 1994,265,808]. Additional homologues of mammalian p38 have been described and include p38p [Jiang et al, J.Biol. Chem, 1996, 271, 17920], p38Y [Li et al, Biochem. Biophys. Res. Commun., 1996, 228, 334] and p388 [jiang et al, J.Biol.Chem. 1997, 272, 30122]. While p38a and p38p are ubiquitously expressed, p38Y is restricted primarily to skeletal muscle and p388 is predominantly expressed in the lung and kidney.
[0010] The release of cytokines by host defensive cells and the response of leukocytes to cytokines and other pro-inflammatory stresses are regulated to varying degrees by p38 MAPK [Cuenda et al., FEBS Lett, 1995, 364, 229-233]. In other cell types, p38 MAPK controls stress responses, such as TNF-α-stimulated IL-8 production by bronchial epithelial cells, and positive regulation of the ICAM-1 cell adhesion molecule in endothelial cells stimulated by LPS. Upon activation, through dual phosphorylation of a TGY motif by dual-specific kinases MKK3 and MKK6, p38 MAPK exerts its effects through phosphorylation of transcription factors and other kinases. MAP kinase-activated protein kinase-2 (MAP-KAPK-2) has been identified as a target for phosphorylation of p38. Mice [Kotlyarov et al., Nat. Cell Biol. 1999, 1, 94-97] lacking MAPKAP-K2 have been shown to release reduced levels of TNF-α, IL-1p, IL-6, IL-10 and IFN-y in response to LPS / galactosamine mediated endotoxic shock. Regulation of the levels of these cytokines as well as COX-2 is at the mRNA level. TNF-a levels are regulated by translational control by AU-rich elements of the 3'-UTR and TNF-a mRNA, with MAPKAP-K2 signaling being the one that increases the translation of TNF-a mRNA. MAPKAP-K2 signaling results in increased stability of COX-2 mRNA, IL-6, and macrophage inflammatory protein. MAPKAP K2 determines the cellular location of MAPK p38 as well as the transduction of MAPK p38 signaling, which has a nuclear localization signal at its carboxyl end and a nuclear export signal as part of its autoinhibitory domain [Engel et al., EMBO J. 1998, 17, 3363-3371]. In stressed cells, MAPKAP-K2 and MAPK p38 migrate to the cytoplasm from the nucleus, this migration only occurring when MAPK p38 is catalytically active. This event is believed to be driven by the exposure of the MAPKAP-K2 nuclear export signal, as a result of phosphorylation by MAPK p38 [Meng et al., J. Biol. Chem. 2002, 277, 37401-37405]. Furthermore, p38 MAPK directly or indirectly leads to the phosphorylation of several transcription factors that are thought to mediate inflammation, including ATF1 / 2 (activating transcription factors 1/2), CHOP-10 / GADD-153 (inducible gene 153 by DNA damage and growth arrest), SAP-1 (serum response factor accessory protein-1) and MEF2C (monocyte enhancing factor-2) [Foster et al., Drug News Perspect. 2000, 13, 488-497].
[0012] In several cases, inhibition of p38 MAPK activity by small molecules has been shown to be useful for the treatment of various pathologies mediated by inadequate cytokine production, including rheumatoid arthritis, COPD, asthma, and cerebral ischemia. This modality has been the subject of several reviews [Salituro et al., Current Medicinal Chemistry, 1999, 6, 807-823 and Kumar et al, Nature Reviews Drug Discovery 2003, 2, 717-726].
[0014] Inhibitors of p38 MAPK have been shown to be effective in animal models of rheumatoid arthritis, such as collagen-induced arthritis in rats [Revesz et al., Biorg. Med. Chem. Lett., 2000, 10, 1261-1364] and adjuvant-induced arthritis in rats [Wadsworth et al., J. Pharmacol. Exp. Ther., 1999, 291,1685-1691. In murine models of pancreatitis-induced lung injury, pretreatment with a MAPK inhibitor p38 reduced airway release of TNF-α and pulmonary edema [Denham et al, Crit. Care Med., 2000, 29, 628 and Yang et al, Surgery, 1999, 126, 216]. Inhibition of p38 MAPK prior to ovalbumin (OVA) exposure in OVA-sensitized mice reduced the accumulation of cytokines and inflammatory cells in the airways in a model of allergic airway inflammation, [Underwood et al., J Pharmacol. Exp. Ther., 2000, 293, 281]. Increased p38 MAP kinase activity has been observed in patients suffering from inflammatory bowel disease [Waetzig et al., J. Immunol, 2002,168, 5432-5351]. Inhibitors of p38 MAPK have been shown to be effective in rat models of cardiac hypertrophy [Behr et al., Circulation, 2001, 104, 1292-1298] and focal cerebral ischemia [Barone et al., J. Pharmacol. Exp. Ther., 2001,296, 312-321].
[0016] The present inventors have discovered a group of compounds which are selective and potent inhibitors of MAPK p38 (p38a, p, y 6) and the isoforms and splice variants thereof, especially p38a, p38p and p38p2. The compounds are therefore useful in medicine, for example, in the treatment and prophylaxis of immune and inflammatory disorders described herein. The compounds are characterized by the presence in the molecule of an amino acid motif or an amino acid ester motif that can be hydrolyzed by an intracellular carboxylesterase. Compounds of the invention having the lipophilic amino acid ester motif cross the cell membrane, and are hydrolyzed in acid by intracellular carboxylesterases. The polar hydrolysis product accumulates in the cell as it does not easily cross the cell membrane. Thus, the compound's p38 MAP kinase activity is prolonged and enhanced within the cell. The compounds of the invention are related to the p38 MAP kinase inhibitors covered by the disclosure in International Patent Application WO03076405, but differ from them in that the present compounds have the amino acid ester motif mentioned above.
[0018] Document WO 2003/076905 deals with monocyclic aroylpyridines, processes for their production and their use as medicines, especially in the treatment of COPD.
[0020] Detailed description of the invention
[0022] According to the invention there is provided a compound of formula (I):
[0027] in which:
[0029] G is -CH =
[0030] D is an optionally substituted phenyl ring;
[0031] R6 is hydrogen;
[0032] P represents hydrogen and U represents a radical of formula (IA); or U represents hydrogen and P represents a radical of formula (IA);
[0034] -A- (CH2) z-X1-L1-Y-NH-CHR1R2 (IA)
[0036] in which
[0038] A represents an optionally substituted phenyl or cyclohexyl ring;
[0039] z is 0 or 1;
[0040] Y is a bond, -C (= O) -, -S (= O) ² -, -C (= O) NRa-, -C (= S) -NRa, -C (= NH) NRa or -S (= O) ² NR3- where R3 is hydrogen or optionally substituted C ¹ -C ⁶ alkyl;
[0042] L1 is a divalent radical of formula - (Alq1) m (Q) n (Alq2) p- in which
[0044] m, n and p are independently 0 or 1,
[0045] Q is (i) a divalent, carbocyclic or heterocyclic, mono or bicyclic, optionally substituted radical having 5-13 ring members, or (ii), in the case where m and p are both 0, a divalent radical of formula -X2- Q1- or -Q1-X2- where X2 is -O-, S- or NRA- where RA is hydrogen or optionally substituted C ¹ -C ³ alkyl, and Q1 is a divalent, carbocyclic or heterocyclic, mono or bicyclic radical , optionally substituted having 5-13 ring members,
[0046] Alk1 and Alq2 independently represent optionally substituted C ³ -C ⁷ cycloalkyl divalent radicals, or alkylene radicals C ¹ -C ⁶ alkenylene or ^C2 - ^C6 alkynylene ^C2 - ^C6 optionally substituted, linear or branched, which may result in, or optionally contain an ether linkage (-O-), thioether (-S -) or amino (-NRA-) where RA is hydrogen or optionally substituted C ¹ -C ³ alkyl; and
[0048] X1 represents a bond; -C (= O); or -S (= O) ² -; -NR4G (= G) -, -C (= O) NR4 -, - NR4C (= O) NR5-, -NR4S (= O) 2- or -S (= O) 2NR4- where R ⁴ and R ⁵ are independently hydrogen or optionally substituted C ¹ -C ⁶ alkyl.
[0049] R ¹ is an ester group that is hydrolyzable by one or more intracellular carboxylesterase enzymes to give a carboxylic acid group; and
[0050] R ² is selected from:
[0052] (i) C ¹ -C ⁶ alkyl, phenyl, 2-, 3- or 4-hydroxyphenyl, 2-, 3- or 4-methoxyphenyl, 2, -3- or 4-pyridylmethyl, benzyl, phenylethyl, 2-, 3- or 4-hydroxybenzyl, 2-, 3- or 4-benzyloxybenzyl, 2-, 3- or 4-C ¹ -C ⁶ alkoxybenzyl, and benzyloxy (C ¹ -C ⁶ alkyl);
[0054] (ii) the characteristic group of a natural amino acid, in which any functional group can be protected;
[0056] (iii) groups - [Alk] n R 6 where Alk is an alkyl group ^(C1 - ^C6) alkenyl or ^(C2 - ^C6) optionally interrupted by one or more -O- or -S- atoms or -N ( R7) - [where R ⁷ is a hydrogen atom or a (C ¹ -C ⁶ ) alkyl group], n is 0 or 1, and R6 is an optionally substituted cycloalkyl or cycloalkenyl group;
[0058] (iv) a benzyl group substituted on the phenyl ring with a group of formula -OCH ² COR ¹⁵ where R ¹⁵ is hydroxyl, amino, (C ¹ -C ⁶ ) alkoxy, phenylalkoxy (C ¹ -C ⁶ ), alkylamino ( C ¹ -C ⁶ ), di ((C ¹ -C ⁶ ) alkyl) amino, phenylalkylamino (C ¹ -C ⁶ ), the remainder of a derivative of an amide, ester, acid halide or amino acid thereof, said residue linked by an amide bond, said amino acid being selected from glycine, a or alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, serine, threonine, cysteine, methionine, asparagine, glutamine, lysine, histidine, arginine, glutamic acid and Aspartic acid;
[0060] (v) a heterocyclicicalkyl group (C ¹ -C ⁶ ), being both unsubstituted and mono or disubstituted on the heterocyclic ring with halo, nitro, carboxy, alkoxy (C ¹ -C ⁶ ), cyano, alkanoyl (C ¹ -C ⁶ ), trifluoromethylalkyl (C ¹ -C ⁶ ), hydroxy, formyl, amino, alkylamino (C ¹ -C ⁶ ), di-alkylamino (C ¹ -C ⁶ ), mercapto, alkylthio (C ¹ -C ⁶ ), hydroxyalkyl ^(C1 - ^C6), mercapto ^(C1 - ^C6) or alquilfenilmetilo ^(C1 - ^C6); and
[0062] (vi) a -CRaRbRc group in which:
[0064] each of Ra, Rb and Rc is independently hydrogen, ^(C1 - ^C6), alkenyl ^(C2 - ^C6), alkynyl ^(C2 - ^C6), phenylalkyl ^(C1 - ^C6), cycloalkyl ( C ³ -C ⁸ ); or
[0066] Rc is hydrogen and Ra and Rb are independently phenyl or heteroaryl, such as pyridyl; or
[0068] Rc is hydrogen, ^(C1 - ^C6), alkenyl ^(C2 - ^C6), alkynyl ^(C2 - ^C6), phenylalkyl ^(C1 - ^C6) alkyl or (C ³ -C ^8), and Ra and Rb together with the carbon atom to which they are attached form a 3 to 8 membered cycloalkyl or a 5 to 6 membered heterocyclic ring; or
[0070] Ra, Rb and Rc together with the carbon atom to which they are attached form a tricyclic ring (for example adamantyl); or
[0072] Ra and Rb are each independently ^(C1 - ^C6), alkenyl ^(C2 - ^C6), alkynyl ^(C2 - ^C6), phenylalkyl (C ¹ -C ^6), or a group as defined for Rc below other than hydrogen, or Ra and Rb together with the carbon atom to which they are attached form a cycloalkyl or heterocyclic ring, and Rc is hydrogen, -OH, -SH, halogen, -CN, -CO ² H, perfluoroalkyl (C ¹ -C ⁴ ), -CH ² OH, -CO ² alkyl (C ¹ -C ⁶ ), -Oalkyl (C ¹ -C ⁶ ), -Oalkenyl (C ² -C ⁶ ), -Salkyl (C ¹ -C ⁶ ), -SO (C ¹ -C ⁶ ) alkyl, -SO ² (C ¹ -C ⁶ ) alkyl, -Salkenyl (C ² -C ⁶ ), -SO (C ² -C ⁶ ) alkenyl, -SO ² alkenyl (C ² -C ⁶ ) or -Q2-W group in which Q ² represents a bond or -O-, -S-, -SO- or -SO ² - and W represents a phenyl, phenylalkyl, cycloalkyl group (C ³ -C ⁸ ), cycloalkylalkyl (C ³ -C ⁸ ), cycloalkenyl (C ⁴ -C ⁸ ), cycloalkenylalkyl (C ⁴ -C ⁸ ), heteroaryl or heteroarylalkyl, group W which may be optionally substituted with one or more selected substituents ind depending on, hydroxyl, halogen, -CN, -CO ² H, -CO ² (C ¹ -C ⁶ ) alkyl, -CONH ² , -CONH (C ¹ -C ⁶ ) alkyl, -CONH (C ¹ -C ⁶ alkyl ) ² , -CHO, -CH ² OH, perfluoroalkyl (C ¹ -C ⁴ ), -Oalkyl (C ¹ -C ⁶ ), -Salkyl (C ¹ -C ⁶ ), -SOalkyl (C ¹ -C ⁶ ), -SO ² (C ¹ -C ⁶ ) alkyl, -NO ² , -NH ² , -NH (C ¹ -C ⁶ ) alkyl, -N ((C ¹ -C ⁶ ) alkyl) ² , -NHCO (C ¹ - C ⁶⁾ alkyl, ^(C1 - ^C6), alkenyl ^(C2 - ^C6), alkynyl ^(C2 - ^C6), cycloalkyl (C ³ -C ⁸⁾ cycloalkyl , (C ⁴ -C ^8), phenyl or benzyl;
[0074] Unless otherwise indicated in the context in which it appears, the term "substituted" as applied to any moiety herein, means substituted with up to four compatible substituents, each of which is independently selected from alkyl ( C ¹ -C ⁶ ), alkoxy (C ¹ -C ⁶ ), hydroxy, hydroxyalkyl (C ¹ -C ⁶ ), mercapto, mercaptoalkyl (C ¹ -C ⁶ ), alkylthio (C ¹ -C ⁶ ), phenyl, halo , including fluorine, bromine and chlorine, trifluoromethyl, trifluoromethoxy, nitro, nitrile (-CN), oxo, -COOH, -COORA, -CORA, -SO ² RA, -CONH ² , -SO ² NH ² , -CONHRA, -SO2NHRA, -CONRARB, -SO2NRARB, -NH2, -NHRa, -NRaRb, -OCONH2, -OCONHRA, -OCONRARB, -NHCORA, -NHCOORA, -NRBCOORA, -NHSO2ORA, -NRBSO2HORA2, -NRBSORHCON2HORA2, -NRBSORHCON2HORA2, -NRBSORHCON2 , -NHCONHRB, -NRACONHRB, -NRACONRARB -NHCONRARB or where RA and RB are independently alkyl ^(C1 - ^C6), cycloalkyl (C ³ -C ^6), phenyl or monocyclic heteroaryl having 5 or 6 ring atoms .
[0075] The compounds of formula (I) above can be prepared in the form of salts, especially pharmaceutically acceptable salts, N-oxides, hydrates and solvates thereof. Any claims for a compound herein, or reference herein to "compounds of the invention", "compounds to which the invention relates", "compounds of formula (I)" and the like, includes salts, N -oxides, hydrates and solvates of such compounds.
[0076] Although the above definition potentially includes high molecular weight molecules, it is preferable, in line with general principles of medical chemistry practice, that the compounds referred to in the present invention have molecular weights of no more than 600.
[0077] The present invention also provides a pharmaceutical composition comprising a compound defined above together with a pharmaceutically acceptable carrier.
[0078] The compounds to which the invention relates can be used for the inhibition of the enzymatic activity of MAP kinase p38 in vitro or in vivo.
[0079] In one aspect of the invention, the invention provides a compound as defined above for the treatment of autoimmune or inflammatory diseases, in particular those mentioned above in which p38 MAP kinase activity is involved.
[0080] Terminology
[0081] The term "ester" or "esterified carboxyl group" means a group RXO (C = O) - in which RX is the characteristic group of the ester, which is theoretically derived from the alcohol RXOH.
[0082] As used herein, the term "(Ca-Cb) alkyl", where a and b are integers, refers to a straight or branched chain alkyl radical having from a to b carbon atoms. Thus, when a is 1 and b is 6, for example, the term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, and n-hexyl. As used herein, the term "divalent alkylene radical (Ca-Cb)", where a and b are integers, refers to a saturated hydrocarbon chain having from a to b carbon atoms and two unsatisfied valences .
[0083] As used herein, the term "alkylene (Ca-Cb)", where a and b are integers, refers to a straight or branched chain alkenyl moiety having from a to b carbon atoms, having at least an E or Z stereochemistry double bond where applicable. The term includes, for example, vinyl, allyl, 1- and 2-butenyl, and 2-methyl-2-propenyl.
[0084] As used herein, the term "divalent alkenylene radical (Ca-Cb)" means a hydrocarbon chain having a to b carbon atoms, at least one double bond, and two unsatisfied valences. As used herein, the term "Ca-Cb alkynyl", where a and b are integers, refers to straight chain or branched chain hydrocarbon groups having from a to b carbon atoms and further having a triple link. This term would include, for example, ethynyl, 1-propynyl, 1- and 2-butynyl, 2-methyl-2-propynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 2-hexynyl, 3-hexynyl, 4- hexynyl and 5-hexynyl.
[0085] As used herein, the term "divalent alkynylene radical (Ca-Cb)", where a and b are integers, refers to a divalent hydrocarbon chain having from a to b carbon atoms, and at least one triple bond.
[0086] As used herein, the term "carbocylic" refers to a mono, bi, or tricyclic radical having up to 16 ring atoms, all of which are carbon, and includes aryl and cycloalkyl.
[0087] As used herein, the term "cycloalkyl" refers to a saturated, monocyclic, carbocyclic radical having 3-8 carbon atoms and includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
[0088] As used herein, the unqualified term "aryl" refers to an aromatic, mono, bi, or tricyclic radical, and includes radicals having two monocyclic carbocyclic aromatic rings that are directly linked by a covalent bond. Illustrative of such radicals are phenyl, biphenyl and naphthyl.
[0089] As used herein, the unqualified term "heteroaryl" refers to a mono, bi, or tricyclic aromatic radical containing one or more heteroatoms selected from S, N, and O, and includes radicals having two such monocyclic rings. , or one such monocyclic ring and a monocyclic aryl ring, which are directly linked by a covalent bond. Illustrative of such radicals are thienyl, benzothienyl, furyl, benzfuryl, pyrrolyl, imidazolyl, benzoimidazolyl, thiazolyl, benzothiazolyl, isothiazolyl, benzoisothiazolyl, pyrazolyl, oxazolyl, benzoxazolyl, isoxazolyl, benzoisoxazolyl, isothiazolyl, pyrrolyloxyazolyl, triazolothiazolyl, triazolyl thiazolyl, triazolyl thiazolyl pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, indolyl, and indazolyl.
[0091] As used herein, the unqualified term "heterocyclyl" or "heterocyclic" includes "heteroaryl" as defined above, and in its non-aromatic meaning refers to a non-aromatic, mono, bi, or tricyclic radical containing one or more heteroatoms selected from S, N and O, and to groups consisting of a monocyclic non-aromatic radical containing one or more of such heteroatoms that is covalently linked to another such radical or to a monocyclic carbocyclic radical. Illustrative of such radicals are pyrrolyl, furanyl, thienyl, piperidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, pyrazolyl, pyridinyl, pyrrolidinyl, pyrimidinyl, piperazinyl, indolyl, morpholinyl, benzfuranyl, pyrazolyl, isoxazolyl, methiazolyl, thiadiazolyl, pyrazolyl, pyridinyl, pyrrolidinyl, pyrimidinyl, piperazinyl, indolyl, morpholinyl, benzfuranyl, pyrazylenyl, isoxazolyl, benzfuranyl, pyrazylenedioranil , maleimido and succinimido.
[0093] A "divalent phenylene, pyridinylene, pyrimidinylene or pyrazinylene radical" is a benzene, pyridine, pyrimidine or pyrazine ring, with two unsatisfied valences and includes 1,3-phenylene, 1,4-phenylene, and the following:
[0098] Unless otherwise specified in the context where it appears, the term "substituted" as applied to any moiety herein means substituted with up to four compatible substituents, each of which may independently be, for example, alkyl ( Ci-Ca), alkoxy (Ci-Ca), hydroxy, hydroxyalkyl (Ci-Ca), mercapto, mercaptoalkyl (Ci-Ca), alkylthio (Ci-Ca), phenyl, halo (including fluorine, bromine and chlorine), trifluoromethyl , trifluoromethoxy, nitro, nitrile (-CN), oxo, -COOH, -COORA, -CORA, -SO ² RA, -CONH ² , -SO ² NH ² , -CONHRA, -SO ² NHRA -CONRARB, -SO ² NRARB, -NH ² , -NHRA, -NRARB, -OCONH ² , -OCONHRA, -OCONRARB, -NHCORA, -NHCOORA, -NRBCOORA, -NHSO ² ORA, -NRBSO ² OH, -NRBSO ² ORA, -NHCONH ² , -NRACONH ² , -NHCONHRB, -NRACONHRB, -NHCONRARB or -NRACONRARB where RA and RB are independently a monocyclic alkyl (Ci-Ca), cycloalkyl (C3-Ca), phenyl or heteroaryl having 5 or more ring atoms. An "optional substituent" can be one of the above substituent groups.
[0100] The term "side chain of a natural or non-natural alpha-amino acid" refers to the group RY in a natural or non-natural amino acid of the formula NH ² -CH (RY) -COOH.
[0102] Examples of natural alpha amino acid side chains include those of alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, histidine, 5-hydroxylysine, 4-hydroxyproline, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, α-aminoadipic acid, α-amino-n-butyric acid, 3,4-dihydroxyphenylalanine, homoserine, α-methylserine, ornithine, pipecolic acid and thyroxine.
[0104] Natural alpha-amino acids that contain functional substituents, for example amino, carboxyl, hydroxy, mercapto, guanidyl, imidazolyl or indolyl groups in their characteristic side chains include arginine, lysine, glutamic acid, aspartic acid, tryptophan, histidine, serine, threonine, tyrosine and cysteine. When R ² in the compounds of the invention is one of said side chains, the optional substituent may be optionally protected.
[0106] The term "protected" when used in relation to a functional substituent on a side chain of a lateral alpha-amino acid means a derivative of such a substituent that is substantially non-functional. For example, carboxyl groups can be esterified (for example as an alkyl Ci-Ca ester), amino groups can be converted to amides (for example as an NHCOalkyl Ci-Ca amide) or carbamates (for example as an NHC (= O) Oalkyl Ci-Ca or NHC (= O) OCH ² Ph carbamate), the hydroxyl groups can be converted to ethers (for example an Oalkyl
[0108] to
[0109] C ¹ -C ⁶ or an O (Ci-C6 alkyl phenyl ether) or esters (for example an OC (= O) alkyl ester) and thiol groups can be converted to thioethers (for example a tert-butyl or benzyl thioether) or thioesters (for example a SC (= O) C ¹ -C ⁶ alkyl thioester). Examples of non-natural alpha-amino acid side chains include those referred to below in the description of R ² groups suitable for use in the compounds of the present invention.
[0111] As used herein, the term "salt" includes base addition, acid addition, and quaternary salts. Compounds of the invention that are acidic can form salts, including pharmaceutically acceptable salts, with bases, such as alkali metal hydroxides, for example sodium and potassium hydroxides; alkaline earth metal hydroxides, for example calcium, barium and magnesium hydroxides; with organic bases, for example, N-methyl-D-glucamine, choline, tris (hydroxymethyl) amino-methane, L-arginine, L-lysine, N-ethylpiperidine, dibenzylamine and the like. Those compounds (I) which are basic can form salts, including pharmaceutically acceptable salts, with inorganic acids, for example with hydric acids, such as hydrochloric or hydrobromic acids, sulfuric acid, nitric acid or phosphoric acid, and the like, and with organic acids. , for example with acetic, tartaric, succinic, fumaric, maleic, malic, salicylic, citric, methanesulfonic, p-toluenesulfonic, benzoic, benzenesulfonic, glutamic, lactic and mandelic acids, and the like. For a review of suitable salts, see Stahl and Wermuth's Handbook of Pharmaceutical Salts: Properties, Selection, and Use (Wiley-VCH, Weinheim, Germany, 2002).
[0113] The term "solvate" is used herein to describe a molecular complex comprising the compound of the invention and a stoichiometric amount of one or more pharmaceutically acceptable solvent molecules, eg, ethanol. The term "hydrate" is used when said solvent is water.
[0115] Compounds of the invention that contain one or more actual or potential chiral centers, due to the presence of asymmetric carbon atoms, can exist as enantiomers or as a variety of diastereoisomers with R or S stereochemistry at each chiral center. The invention includes all of these enantiomers and diastereoisomers and mixtures thereof.
[0117] As mentioned, the esters of the invention are converted by intracellular esterases to carboxylic acids. Both esters and carboxylic acids can have p38 MAP kinase inhibitory activity in their own right.
[0119] In the compounds referred to by the invention:
[0121] Group D
[0122] D is an optionally substituted phenyl ring.
[0123] The R6 substituent
[0124] R6 is hydrogen.
[0125] Regioisomers P / U
[0126] At present it is preferred that P is hydrogen and U is a radical of formula (IA) as defined above.
[0127] The radical A
[0128] In the radical of formula (IA), it is presently preferred that A is optionally substituted 1,4-phenylene. In that case, preferred optional substituents include fluorine and chlorine. A can also be a cyclohexyl ring, which is optionally substituted.
[0130] A particularly preferred subgroup of compounds of the invention consists of those of formula (IIA), (IIB) and (IIC):
[0133] where R ¹¹ = F, R ¹² = H, R ¹³ = H and R ¹⁴ = H; or
[0134] R ¹¹ = F, R ¹² = F, R ¹³ = H and R ¹⁴ = H; or
[0135] R ¹¹ = F, R ¹² = H, R ¹³ = F and R ¹⁴ = F; or
[0136] R ¹¹ = F, R ¹² = F, R ¹³ = F and R ¹⁴ = F; or
[0137] R ¹¹ = F, R ¹² = F, R ¹³ = F and R ¹⁴ = H
[0139] and those where z, X1, L1, Y, R1 and R2 are as defined above with reference to formula (I), and as further described below.
[0141] The radical -YL ¹ -X ¹ - [CH2] ^z -
[0142] This radical (or bond) arises from a particular chemical strategy selected for attaching the R ¹ CH (R ² ) NH- amino acid ester motif to the A ring system. Obviously, the chemical strategy for that coupling can vary widely, and hence therefore many combinations of the variables Y, L1, X1 and z are possible. The precise combination of variables that make up the binding chemistry between the amino acid ester motif and the A ring system will usually be irrelevant to the primary binding mode of the compound as a whole. On the other hand, such binding chemistry will in some cases pick up additional binding interactions with the enzyme. It should also be noted that the benefits of the amino acid ester motif (easy entry into the cell, hydrolysis of esterase within the cell, and accumulation within the cell of active carboxylic acid hydrolysis product) are best achieved when Bonding between the amino acid ester motif and the A ring system is not a substrate for peptidase activity within the cell, which can result in cleavage of the amino acid from the molecule. Of course, stability for intracellular peptidases is easily tested by incubating the compound with discontinuous cell contents, and testing for any such cleavages.
[0144] With the above general observations in mind, taking in turn the variables that constitute the radical -Y-L1-X1- [CH2] z-:
[0146] z can be 0 or 1, so that a methylene radical attached to ring system A is optional;
[0148] specific examples of Y when macrophage selectivity is not required include - (C = O) - and - (C = O) NH-; Where selectivity of a macrophage is required, any of the other options for Y are suitable, including the case where Y is a bond.
[0149] In the radical L1, examples of radicals Alq1 and Alq2, when present, include -CH ² -, -CH ² CH ² -, -CH ² CH ² CH ² -, -CH ² CH ² CH ² CH ² -, -CH = CH-, -CH = CHCH ² -, -CH ² CH = CH-, CH ² CH = CHCH ² -C = C-, -C = CCH ² -, CH ² C = C- and CH ² C = CCH ² . Additional examples of Alq1 and Alq2 include -CH ² W-, -CH ² CH ² W-, -CH ² CH ² WCH ² -, -CH2CH2WCH (CH3) -, -CH ² WCH ² CH ² -, -CH ² WCH ² CH ² WCH ² - and -WCH ² CH ² -, where W is -O-, -S-, -NH-, -N (CH3) - or -CH ² CH ² N (CH ² CH ² OH) CH ² -. Additional examples of Alq1 and Alq2 include divalent cyclopropyl, cyclopentyl and cyclohexyl radicals.
[0150] In L1, when n is 0, the radical is a hydrocarbon chain (optionally substituted and perhaps having an ether, thioether, or amino linkage). Currently, it is preferred that there are no optional substituents on L1. When m and p are both 0, L1 is a divalent carbocyclic or heterocyclic, monocyclic or bicyclic radical with 5-13 ring atoms (optionally substituted). When n is 1 and at least one of m and p is 1, L1 is a divalent radical that includes a hydrocarbon chain or chains and a carbocyclic or heterocyclic, mono or bicyclic radical with 5-13 ring atoms (optionally substituted). When present, Q can be, for example, a divalent phenyl, naphthyl, cyclopropyl, cyclopentyl or cyclohexyl radical, or a heterocyclic, mono or bicyclic radical which It has 5 to 13 ring members, such as a piperidinyl, piperazinyl, indolyl, pyridyl, thienyl or pyrrolyl radical, but 1,4-phenylene is presently preferred.
[0151] Specifically, in some embodiments of the invention, L1, m and p can be 0, where n is 1. In other embodiments, n and p can be 0, where m is 1. In other embodiments, m, n and p can all be 0. In still additional embodiments m can be 0, n can be 1, where Q is a monocyclic heterocyclic radical, and p can be 0 or 1. Alq1 and Alq2, when present, can be selected from -CH ² -, -CH ² CH ² - and -CH ² CH ² CH ² - and Q can be 1,4-phenylene.
[0153] Specific examples of the radical -Y-L1-X1- [CH ² ] z- include -C (= O) - and -C (= O) NH-, as well as - (CH ² ) v-, - (CH ² ) vO-, -C (= O) - (CH ² ) v-, -C (= O) - (CH ² ) vO-, -C (= O) -MH- (CH2) w-, -C ( = O) -NH- (CH ² ) wO-
[0158] where v is 1, 2, 3 or 4 and w is 1, 2 or 3, such as -Y-L1-X1- [CH ² ] z-, is -CH ² -, -CH ² CH ² -, - CH ² CH ² CH ² -, -CH2CH2CH2CH2-, -CH2O-, -CH2CH2O-, -CH2CH2CH2O-, -CH2CH2CH2CH2O-, -C (= O) -CH2-, -C (= O) -CH2O-, -C (= O) -NH-CH ² - or -C (= O) -NH-CH ² O-.
[0160] The Ri group
[0162] The ester group R ¹ must be one that in the compound of the invention is hydrolyzable by one or more intracellular carboxylesterase enzymes to give a carboxylic acid group. Intracellular carboxylesterase enzymes capable of hydrolyzing the ester group of a compound of the invention to give the corresponding acid include the three known human enzyme isotopes hCE-1, hCE-2 and hCE-3. Although these are considered to be the main enzymes, other enzymes, such as biphenylhydrolase (BPH) can also play a role in the hydrolysis of the ester. In general, if the carboxylesterase hydrolyzes the free amino acid ester to give the parent acid, it will also hydrolyze the ester motif when covalently conjugated to the inhibitor. Thus, the broken cell assay and / or isolated carboxylesterase assay described herein provide a simple, rapid, and direct first screen for esters having the required hydrolysis profile. The ester motifs selected in such a way can then be retested in the same carboxylesterase assay when conjugated to the inhibitor by the selected conjugation chemistry, to confirm that it is still a carboxylesterase substrate at that background level.
[0164] Subject to the requirement that they be hydrolyzed by intracellular carboxylesterase enzymes, examples of particular R ¹ ester groups include those of formula - (C = O) OR14 where R ¹⁴ is R ⁸ R ⁹ R ¹⁰ C- in which
[0166] (i) R8 is hydrogen or (C1-C3) alkyl - (Z1) a- [(C1-C3) alkyl] b- optionally substituted or (C2-C3) alkenyl - (Z1) a- [(C1-C3) alkyl )] b- where a and b are independently 0 or 1 and Z1 is -O-, -S- or -NR ¹¹ -where R ¹¹ is hydrogen or (C ¹ -C ³ ) alkyl; and R ⁹ and R ¹⁰ are independently hydrogen or (C ¹ -C ³ ) alkyl -;
[0168] (ii) R8 is hydrogen or R12R13N-(C ¹ -C ³ ) alkyl - optionally substituted where R ¹² is hydrogen or (C ¹ -C ³ ) alkyl and R ¹³ is hydrogen or (C ¹ -C ³ ) alkyl; or R ¹² and R ¹³ together with the nitrogen to which they are attached form an optionally substituted monocyclic heterocyclic ring of 5 or 6 ring atoms or a bicyclic heterocyclic ring system of 8 to 10 ring atoms, and R ⁹ and R ¹⁰ are independently hydrogen or (C1-C3) alkyl -; or
[0169] (iii) R8 and R ⁹ , taken together with the carbon to which they are attached, form an optionally substituted monocyclic carbocyclic ring of 3 to 7 ring atoms or bicyclic carbocyclic ring system of 8 to 10 ring atoms, and R ¹⁰ is hydrogen.
[0171] Within these classes, R ¹⁰ is usually hydrogen. Specific examples of R ¹⁴ include methyl, ethyl, n- or iso-propyl, n-, sec- or tert-butyl, cyclohexyl, allyl, phenyl, benzyl, 2-, 3- or 4-pyridylmethyl, N-methylpiperidine- 4-yl, tetrahydrofuran-3-yl or methoxyethyl. It is presently preferred where R ¹⁴ is cyclopentyl.
[0173] Macrophages are known to play a key role in inflammatory disorders through the release of cytokines, in particular TNF-a and IL-1 (van Roon et al., Arthritis and Rheumatism, 2003, 1229-1238). In rheumatoid arthritis, they contribute primarily to the maintenance of joint inflammation and the destruction of the joint. Macrophages are also involved in tumor growth and development (Naldini and Carraro, Curr Drug Targets Inflamm Allergy, 2005, 3-8). Therefore, agents that selectively target macrophage cell proliferation could be of value for the treatment of cancer and autoinflammatory diseases. Targeting specific cell types might be expected to lead to a reduction in side effects. The inventors have discovered a method of targeting p38 kinase inhibitors to macrophages that is based on the observation that how the esterase motif binds to the p38 kinase inhibitor determines whether it is hydrolyzed, and therefore whether or not it accumulates in different cell types. Specifically, macrophages have been found to contain the human hCE-1 carboxyesterase, while other cell types do not.
[0174] In the general formula (I), when the nitrogen of the esterase motif R-iCH (R ² ) NH- is not directly attached to a carbonyl (-C (= O) -), that is, when Y is not a radical -C (= O), -C (= O) O- or -C (= O) NR3-, the ester will be hydrolyzed only by hCE-1 and therefore the inhibitors will only accumulate in macrophages. Unless "monocyte" or "monocytes" are specified herein, the term macrophage or macrophages will be used to denote macrophages (including tumor associated macrophages) and / or monocytes.
[0176] The R2 amino acid side chain
[0178] Subject to the requirement that the R ¹ ester group be hydrolyzed by intracellular carboxylesterase enzymes, the identity of the R ² side chain group is not critical.
[0180] They are amino acid side chains:
[0182] C ¹ -C ⁶ alkyl, phenyl, 2-, 3- or 4-hydroxyphenyl, 2-, 3- or 4-methoxyphenyl, 2, -3- or 4-pyridylmethyl, benzyl, phenylethyl groups,
[0183] 2-, 3- or 4-hydroxybenzyl, 2-, 3- or 4-benzyloxybenzyl, 2-, 3- or 4-C ¹ -C ⁶ alkoxybenzyl, and benzyloxy (C ¹ -C ⁶ alkyl) -;
[0184] the characteristic group of a natural amino acid, in which any functional group can be protected;
[0186] groups - [Alk] n R 6 where Alk is an alkyl group ^(C1 - ^C6) alkenyl or ^(C2 - ^C6) optionally interrupted by one or more -O- or -S- atoms or -N (R7) - [where R ⁷ is a hydrogen atom or a (C ¹ -C ⁶ ) alkyl group], n is 0 or 1, and R6 is an optionally substituted cycloalkyl or cycloalkenyl group;
[0188] a benzyl group substituted on the phenyl ring with a group of formula -OCH ² COR ¹⁵ where R ¹⁵ is hydroxyl, amino, alkoxy (C ¹ -C ⁶ ), phenylalkoxy (C ¹ -C ⁶ ), alkylamino (C ¹ -C ⁶ ), di (C ¹ -C ⁶ alkyl) amino, phenylalkylamino (C ¹ -C ⁶ ), the residue of an acid halide or amino acid, derived from an ester or amide thereof, said residue being linked by a bond of amide, said amino acid being selected from glycine, a or alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, serine, threonine, cysteine, methionine, asparagine, glutamine, lysine, histidine, arginine, glutamic acid and aspartic acid;
[0190] a heterocyclic (C ¹ -C ⁶ ) alkyl group, being both unsubstituted and mono or disubstituted on the heterocyclic ring with halo, nitro, carboxy, alkoxy (C ¹ -C ⁶ ), cyano, alkanoyl (C ¹ -C ⁶ ), trifluoromethyl (C ¹ -C ⁶ ) alkyl, hydroxy, formyl, amino, alkylamino (C ¹ -C ⁶ ), di-alkylamino (C ¹ -C ⁶ ), mercapto, alkylthio (C ¹ -C ⁶ ), hydroxyalkyl (C ¹ -C ⁶ ), mercaptoalkyl
[0191] (C ¹ -C ⁶ ) or (C ¹ -C ⁶ ) alkylphenylmethyl; and
[0193] a -CRaRbRc group in which:
[0195] each of Ra, Rb and Rc is independently hydrogen, ^(C1 - ^C6), alkenyl ^(C2 - ^C6), alkynyl ^(C2 - ^C6), phenylalkyl ^(C1 - ^C6), cycloalkyl ( C ³ -C ⁸ ); or
[0197] Rc is hydrogen and Ra and Rb are independently phenyl or heteroaryl such as pyridyl; or
[0199] Rc is hydrogen, ^(C1 - ^C6), alkenyl ^(C2 - ^C6), alkynyl ^(C2 - ^C6), phenylalkyl ^(C1 - ^C6) alkyl or (C ³ -C ^8), and
[0200] Ra and Rb, together with the carbon atom to which they are attached, form a 3 to 8 membered cycloalkyl or a 5 to 6 membered heterocyclic ring; or
[0202] Ra, Rb and Rc, together with the carbon atom to which they are attached, form a tricyclic ring (for example adamantyl); or
[0204] Ra and Rb are each independently ^(C1 - ^C6), alkenyl ^(C2 - ^C6), alkynyl ^(C2 - ^C6), phenylalkyl (C ¹ -C ⁶⁾ or a group as defined for Rc below other than hydrogen, or Ra and Rb, together with the carbon atom to which they are attached, form a cycloalkyl or heterocyclic ring, and Rc is hydrogen, -OH,
[0205] -SH, halogen, -CN, - CO ² H, perfluoroalkyl (C ¹ -C ⁴ ), -CH ² OH, -CO ² alkyl (C ¹ -C ⁶ ), -Oalkyl (C ¹ -C ⁶ ), - -Oalkenyl (C ² -C ⁶ ), -Salkyl (C ¹ -C ⁶ ), -SOalkyl (C ¹ -C ⁶ ), -SO ² alkyl (C ¹ -C ⁶ ), -Salkenyl (C ² -C ⁶ ) , -SOalkenyl (C ² -C ⁶ ), -SO ² alkenyl (C ² -C ⁶ ) or a -Q2-W group, where Q2 represents a bond or -O-, -S-,
[0206] -SO- or -SO ² - and W represents a group phenyl, phenylalkyl, cycloalkyl (C ³ -C ⁸ ), cycloalkylalkyl (C ³ -C ⁸ ), cycloalkenyl (C ⁴ -C ⁸ ), cycloalkenylalkyl (C ⁴ -C ⁸ ), heteroaryl or heteroarylalkyl, group W which may be optionally substituted with one or more substituents independently selected from, hydroxyl, halogen, -CN, -CO ² H, -CO ² (C ¹ -C ⁶ ) alkyl, -CONH ² , -CONHalkyl (C ¹ -C ⁶ ), -CONH (C ¹ -C ⁶ alkyl) ² , -CHO, -CH ² OH, perfluoroalkyl (C ¹ -C ⁴ ), -Oalkyl (C ¹ -C ⁶ ), -Salkyl (C ¹ -C ⁶ ), -SOalkyl (C ¹ -C ⁶ ), -SO ² alkyl (C ¹ -C ⁶ ),
[0207] -NO ² , -NH ² , -NH (C ¹ -C ⁶ ) alkyl, -N ((C ¹ -C ⁶ ) alkyl) ² , -NHCO (C ¹ -C ⁶ ) alkyl, (C ¹ -C ⁶ ) alkyl , alkenyl (C ² -C ⁶ ), cycloalkyl (C ³ -C ⁸ ), cycloalkenyl (C ⁴ -C ⁸ ), phenyl or benzyl.
[0209] Particular examples of R ² groups include hydrogen (the glycine "side chain"), benzyl, phenyl, cyclohexylmethyl, cyclohexyl, pyridin-3-ylmethyl, tert-butoxymethyl, iso-butyl, sec-butyl, tert-butyl, 1 -benzylthio-1-methylethyl,
[0210] 1-methylthio-1-methylethyl, 1-mercapto-1-methylethyl and phenylethyl. Currently, preferred R ² groups include phenyl, benzyl, isobutyl, cyclohexyl, and t-butoxymethyl.
[0212] For the compounds of the invention to be administered systemically, esters with a low rate of carboxylesterase cleavage are preferred, as they are less susceptible to pre-systemic metabolism. Therefore, its ability to reach its target tissue intact is increased, and the ester can be converted to the acid product inside the cells of the target tissue. However, for local administration, where the ester is applied directly to or targeted to the target tissue by, for example, inhalation, it will often be desirable for the ester to have a high rate of cleavage by esterases, to minimize systemic exposure. and the consequent unwanted side effects. In the compounds of the present invention, if the carbon adjacent to the alpha carbon of the alpha amino acid ester is monosubstituted, that is, R ² is CH ² Rz (with Rz being the mono-substituent) the esters tend to cleave more rapidly than if that carbon was di or trisubstituted, as in the case where R ² is, for example, phenyl or cyclohexyl.
[0214] As mentioned above, the compounds to which the invention relates are inhibitors of the activity of MAP kinase p38, and therefore are useful in the treatment of diseases, such as psoriasis, inflammatory bowel disease, Crohn's disease , ulcerative colitis, chronic obstructive pulmonary disease, asthma, multiple sclerosis, diabetes, atopic dermatitis, graft-versus-host disease, or systemic lupus erythematosus and rheumatoid arthritis, in which the activity of MAP kinase p38 is involved.
[0216] It will be understood that the specific dosage level for any particular patient will depend on a variety of factors including the activity of the specific compound employed, age, body weight, general health, sex, diet, time. of administration, the route of administration, the rate of excretion, the combination of drugs, and the severity of the particular disease undergoing treatment. Optimal dosage levels and dosing frequency will be determined by clinical trial.
[0218] The compounds to which the invention relates can be prepared for administration by any route consistent with their pharmacokinetic properties. Orally administrable compositions may be in the form of tablets, capsules, powders, granules, lozenges, liquid or gel preparations, such as sterile oral, topical or parenteral solutions or suspensions. Tablets or capsules for oral administration may be in unit dose presentation form, and may contain conventional excipients, such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone; fillers, for example, lactose, sugar, cornstarch, calcium phosphate, sorbitol, or glycine; tablet lubricant, eg, magnesium stearate, talc, polyethylene glycol, or silica; disintegrants, for example potato starch, or suitable wetting agents, such as sodium lauryl sulfate. The tablets can be coated according to methods well known in normal pharmaceutical practice. Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or they may be presented as a dry product for constitution with water or other suitable vehicle before use. Said liquid preparations may contain conventional additives, such as suspending agents, for example, sorbitol, syrup, methyl cellulose, glucose syrup, gelatin, hydrogenated edible fats; emulsifying agents, for example, lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which may include edible oils), for example, almond oil, fractionated coconut oil, oily esters, such as glycerin, propylene glycol, or ethyl alcohol; preservatives, for example methyl or propyl p-hydroxybenzoate or sorbic acid, and if desired conventional flavoring or coloring agents.
[0220] For topical application to the skin, the drug can be made into a cream, lotion, or ointment. Cream or ointment formulations that can be used for the drug are conventional formulations known in the art, for example, as described in conventional textbooks of pharmacy, such as the British Pharmacopoeia.
[0221] For topical application by inhalation, the drug may be formulated, for example, for aerosol administration, by pressure-directed jet atomizers or ultrasonic atomizers, or preferably by propellant-driven metered aerosols or the non-propellant administration of micronized powders, for example , inhalation capsules or other "dry powder" delivery systems. Excipients, such as, for example, propellants (for example, frigene in the case of metered aerosols), surfactants, emulsifiers, stabilizers, preservatives, flavors, and fillers (for example, lactose in the case of powder inhalers) can be present in such inhaled formulations. For inhalation purposes, a wide variety of apparatuses are available with which aerosols of optimal particle size can be generated and administered, using an inhalation technique that is suitable for the patient. In addition to the use of adapters (spacers, expanders) and pear-shaped containers (e.g. Nebulator®, Volumatic®), and automatic devices that emit a spray jet (Autohaler®), for metered aerosols, particularly in the In the case of powder inhalers, a number of technical solutions are available (eg Diskhaler®, Rotadisk®, Turbohaler® or the inhalers described, for example, in European Patent Application EP 0505321).
[0223] For topical application to the eye, the drug can be prepared in a solution or suspension in a suitable sterile aqueous or non-aqueous vehicle. Additives may also be included, for example buffers such as sodium metabisulfite or disodium edeatate; preservatives, including fungicidal and bactericidal agents, such as phenylmercuric acetate or nitrate, benzalkonium chloride or chlorhexidine, and thickening agents, such as hypromellose.
[0224] The active ingredient can also be administered parenterally in a sterile environment. Depending on the vehicle and concentration used, the drug can be suspended or dissolved in the vehicle. Advantageously, adjuvants, such as a local anesthetic, preserving agents, and buffers can be dissolved in the vehicle.
[0226] Synthesis
[0228] There are multiple synthetic strategies for the synthesis of compounds (I) with regard to the present invention, but all rely on known chemical procedures, known to the specialist in synthetic organic chemistry. Therefore, compounds according to formula (I) can be synthesized according to procedures described in conventional literature and are well known to those skilled in the art. Typical bibliographic sources are "Advanced organic chemistry", 4th edition (Wiley), J March, "Comprehensive Organic Transformation", 2nd Edition (Wiley), RC Larock, "Handbook of Heterocyclic Chemistry", 2nd edition (Pergamon), AR Katritzky) , review articles such as those found in "Synthesis", "Acc. Chem. Res.", "Chem. Rev", or primary bibliographic sources identified by online searches of conventional bibliography or secondary sources, such as "Chemical Abstracts " or " Beilstein ".
[0230] Compounds of the invention can be prepared by a variety of procedures that are described generally below and more specifically in the Examples hereinafter. In the reactions described below, it may be necessary to protect reactive functional groups, eg, hydroxyl, amino and carboxy groups, where these are desired in the final product, to prevent their unwanted participation in the reactions [see for example Greene, TW, "Protecting Groups in Organic Synthesis", John Wiley and Sons, 1999]. Conventional protecting groups can be used in conjunction with conventional practice. In some cases, deprotection may be the final step in the synthesis of a compound of the general formula (I) and it is understood that the processes according to the invention described later in the present document extend to said removal of protecting groups .
[0232] Examples of such methods, which can be used for the synthesis of compounds of the general formula (I) are set forth, but not limited to, in the reactions shown below in Scheme 1.
[0233] Scheme 1
[0241] Thus, aminoesters of general formula (A) can be prepared by treating tert-butyl carbamate of general formula (2a) with trifluoroacetic acid in dichloromethane. Intermediates of general formula (2) can be prepared by methods described in WO 03/076405 and references therein. The amino esters of general formula (2b) can be formed as a by-product in the synthesis of compounds of formula (2a) and treated with trifluoroacetic acid to give compounds of general formula (B).
[0243] Intermediate esters of general formula (5) can be prepared by the procedures shown in Scheme 2.
[0244] Scheme 2
[0249] Hydrogenation of the nitrobenzyl intermediate (6) over palladium-carbon catalyst in THF provides amines of the general formula (5). Intermediates of formula (6) can be prepared by reacting the corresponding amine with di-tert-butoxycarbonate in an inert solvent, such as THF at room temperature. Intermediates of the general formula (7) can be produced by alkylating aminoesters of formula (8) with 4-nitrobenzyl bromide. The reaction can be carried out in a dialkylamide solvent, such as DMF in the presence of an inorganic base, such as potassium or cesium carbonate. Such reactions are discussed in March's Advanced Organic Chemistry [John Wiley and Sons, 1992].
[0251] An alternative method for the synthesis of N-benzylamino acid esters of general formula (9), where further purification is required at the aryl ring of the benzyl substituent, is set forth in Scheme 3.
[0256] In a further aspect of the invention, the aminoesters of general formula (9) can be prepared, but not limited to, by the reactions set forth in Scheme 3. Thus, benzonitriles of general formula (11), which are commercially available or can be easily synthesized by methods known to those skilled in the art, they can be converted to the corresponding benzaldehyde of general formula (10) by reduction with a suitable metal hydride, such as DIBAL-H and acid hydrolysis of the intermediate imine [see, for example LeBel J. Am. Chem. Soc., 1964, 86, 3759]. The N-benzyl amino acid ester of general formula (9) can be prepared by reaction with the mentioned benzaldehyde under reductive alkylation conditions, using borohydride reagents, such as NaBHaCN or NaBH (OAc) 3, under acidic conditions, in a protic solvent. , such as methanol [see, for example, Borsch et al, J.Am. Chem. Soc., 1971,93, 2897].
[0257] Scheme 4
[0265] In a further aspect of the invention, compounds of general formula (C) can be prepared by methods set forth in Scheme 4, starting from the alkylation of intermediates of general formula (8) with mesylates of general formula (12). The alkylation can be carried out in an inert solvent, such as THF, in the presence of sodium iodide and inorganic bases, such as potassium carbonate. It will be recognized by those skilled in the art that the corresponding alkyl bromides or alkylchlorides will be useful in this process. The preparation of the mesylate (12) can be carried out by treating the primary alcohol (13) with methanesulfonyl chloride, in an inert solvent, such as dichloromethane and in the presence of an organic base, such as triethylamine. Compounds of general formula (14) can be prepared by methods described in WO 03/076405 and references therein.
[0267] In a further aspect of the invention, compounds of general formula (D) can be prepared, but not limited to, by the reactions in Scheme 5.
[0270] Therefore, alcohols of general formula (14) can be alkylated with a suitably protected cycloalkanol derivative, such as 1,4-dioxaspiro [4,5] decan-8-ol using triphenylphosphine and a dialkyl azadicarboxylate, such as DEAD in an inert ethereal solvent [see, eg, Mitsunobu et al, Bull. Chem. Soc. Jpn., 1967, 40, 2380]. Ketals of general formula (16) can be protected to give the corresponding ketone under acidic aqueous conditions. Reductive amination of compounds of formula (16) can be achieved by treatment with amino acid esters of the general formula (8) in the presence of borohydride reagents, such as sodium cyanoborohydride and sodium triacetoxyborohydride under acidic conditions to give compounds of the general formula ( D).
[0272] In a further aspect for the invention, compounds of general formula (E) can be prepared, but not limited to, by the reactions in Scheme 6.
[0277] Reductive amination of compounds of formula (22) can be achieved by treatment with dibenzylamine in the presence of borohydride reagents, such as sodium cyanoborohydride and sodium triacetoxyborohydride under acidic conditions to give compounds of general formula (21). Hydrogenation of (21) and subsequent reaction with thiophosgene can give the isocyanate of general formula (20). Compounds of the general formula (19) can be prepared by reacting (20) with the corresponding acetophenone using sodium tert-butoxide. The alkylation of (19) with iodoethane can be carried out using an inorganic base, such as potassium carbonate in a solvent, such as acetone. Compounds of the general formula (18) can undergo cyclization, oxidation and then subsequent ammonium displacement to give the ketals of the general formula (17). Thus, the ketals of general formula (17) can be deprotected to give the corresponding cyclohexanone intermediate under acidic aqueous conditions, then cyclohexanone reacts with amino acid esters of general formula (8) under reductive amination conditions using borohydride reagents, such as sodium cyanoborohydride and sodium triacetoxyborohydride.
[0279] In another aspect of the invention, amino acids of general formula (F) can be prepared by, but not restricted to, the methods set forth in Scheme 7.
[0280] Scheme 7
[0285] Thus, for example, amino acid esters of the general formula (C) can be hydrolyzed to the corresponding amino acids (F) by treatment with aqueous sodium or potassium hydroxide, or any suitable base, at room temperature in a cosolvent, such like methanol or ethanol.
[0287] In another aspect of the invention, amino acids of general formula (G) can be prepared by, but not restricted to, the methods set forth in Scheme 8.
[0289] Scheme 8
[0294] Therefore, aminoesters of general formula G can be prepared by alkylating intermediates of general formula (8) with mesylates of general formula (23). The alkylation can be carried out in an inert solvent, such as THF, in the presence of sodium iodide and inorganic bases, such as potassium carbonate. The preparation of Mesylate (23) can be made by treating the primary alcohol (24) with methanesulfonyl chloride in an inert solvent, such as dichloromethane and in the presence of an organic base, such as triethylamine. Alcohol (24) can be prepared by deprotection of the acetyl group of intermediate (25) under acidic conditions, such as HCl. Intermediates of general formula (4), (25) and (26) can be prepared by similar methods described in WO 03/076405 and references therein.
[0296] The following examples illustrate the invention. All temperatures are in ° C. The following abbreviations are used:
[0297] MeOH = methanol
[0298] EtOH = ethanol
[0299] EtOAc = ethyl acetate
[0300] Boc = tert-butoxycarbonyl
[0301] CDI = 1,1'-carbonyldiimidazole
[0302] DCM = dichloromethane
[0303] DMF = dimethylformamide
[0304] DMSO = dimethylsulfoxide
[0305] TFA = trifluoroacetic acid
[0306] THF = tetrahydrofuran
[0307] Na2CO3 = sodium carbonate
[0308] HCl = hydrochloric acid
[0309] DIPEA = diisopropylethylamine
[0310] NaH = sodium hydride
[0311] NaOH = sodium hydroxide
[0312] NaHCO3 = sodium hydrogen carbonate
[0313] Pd / C = palladium on carbon
[0314] TME = tert-butyl methyl ether
[0315] N ² = nitrogen
[0316] Na2SO4 = sodium sulfate
[0317] EtsN = triethylamine
[0318] NH ³ = ammonia
[0319] TMSCl = trimethylchlorosilane
[0320] TBME = tertiary butyl methyl ether
[0321] NH4Cl = ammonium chloride
[0322] LiAlH4 = lithium aluminum hydride
[0323] MgSO4 = magnesium sulfate
[0324] nBuLi = n-butyllithium
[0325] CO ² = carbon dioxide
[0326] EDCl = W- (3-dimethylaminopropyl) -W'-ethylcarbodiimide hydrochloride
[0327] Et2O = diethyl ether
[0328] LiOH = lithium hydroxide
[0329] HOBt = 1-hydroxybenzotriazole
[0330] ELS = Evaporative Light Scattering
[0331] TLC = thin layer chromatography
[0332] ml = milliliter (s)
[0333] g = gram (s)
[0334] mg = milligrams (s)
[0335] mole = moles
[0336] mmol = millimole (s)
[0337] LCMS = high performance liquid chromatography / mass spectrometry
[0338] NMR = nuclear magnetic resonance
[0339] TA = room temperature
[0341] Microwave irradiation was performed using a CEM Discover directed microwave reactor. Solvents were removed using a GeneVac Series I without heating or a Genevac Series
[0343] II with VacRamp at 30 ° C or a Buchi rotary evaporator. Purification of compounds by flash column chromatography was performed using silica gel, particle size 40-63 pm (230-400 mesh) obtained from Silicicle. Compound purification by preparative HPLC was performed on Gilson systems using ThermoHypersil-Keystone Hyperprep HS C18 reversed phase columns (12 pm, 100 x 21.2 mm), 20-100% B gradient (A = water / 0 TFA , 1%, B = acetonitrile / 0.1% TFA) for 9.5 min, flow = 30 ml / min, DMSO 2: 1 injection solvent: acetonitrile (1.6 ml), UV detection at 215 nm .
[0345] 1 H NMR spectra were recorded on a Bruker 400 MHz AV spectrometer or a Bruker 300 MHz AV spectrometer in deuterated solvents. Chemical shifts (8) are in parts per million. Thin layer chromatography (TLC) analysis was performed with Kieselgel 60 F ²⁵⁴ plates (Merck) and visualized using UV light.
[0346] Analytical HPLCMS was performed on Agilent HP1100, Waters 600 or Waters 1525 LC systems using Hypersil BDS C18 reverse phase columns (5 µm, 2.1 x 50 mm), 0-95% gradient B (A = water / TFA 0.1%, B = acetonitrile / 0.1% TFA) for 2.10 min, flow = 1.0 ml / min. UV spectra were recorded at 215 nm using a Gilson G1315A diode array detector, G1214A single wavelength UV detector, Waters 2487 dual wavelength UV detector, Waters 2488 dual wavelength UV detector, Array UV detector. of Waters 2996 diodes. Mass spectra were obtained in the m / z range 150 to 850, at a sampling rate of 2 scans per second or 1 scan every 1.2 seconds using a Micromass LCT with Z-spray interface o Micromass LCT with Z-spray or MUX interface. Data was integrated and reported using OpenLynx and OpenLynx Browser software.
[0348] Intermediate
[0350] Intermediate 1A (S) -2-r (4-Am¡nobenz¡n-tert-butoxycarbonylamino1-4-cyclopentyl methylpentanoate
[0355] Cyclopentyl (S) -2- [tert-butoxycarbonyl- (4-nitrobenzyl) amino] -4-methylpentanoate (3.8 g, 8.74 mmol) was dissolved in EtOH (100 mL) prior to the addition of catalyst Pd / C (10% humidity) (100 mg) and hydrogenated under balloon pressure at room temperature for 18 h. The reaction mixture was filtered through a pad of celite and evaporated to dryness to give a pink solid (3.15 g, 89% yield). LCEM purity 100%, m / z 405 [M + H] +.
[0356] The benzyl carbamate starting material for this procedure was prepared as follows:
[0361] Cyclopentyl (S) - (4-nitrobenzyl) amino] -4-methylpentanoate (15.8 g, 47.4 mmol) was dissolved in THF (250 mL) prior to addition of potassium carbonate (7.58 g, 56 , 9 mmol) and water (150 ml). Di-tert-butyl dicarbonate (15.5 g, 71.1 mmol) was added and the reaction mixture was heated at 50 ° C for 18 h. The reaction mixture was concentrated under reduced pressure to remove volatiles, giving an aqueous residue that was extracted with EtOAc (200 ml). The EtOAc layer was washed consecutively with 0.1M HCl (150 mL), sat aq NaHCO3. and water (150 ml). The organic layer was dried (Na2SO4), filtered, and concentrated to dryness. After purification by flash column chromatography (10% EtOAc / hexane) the product was isolated (9.36 g, 46% yield). LC 94% purity, m / z 435 [M + H] +.
[0363] The nitrobenzylamino starting material used in this procedure was prepared as follows
[0365] 4-Nitrobenzyl bromide (11 g, 50 mmol) was dissolved in DMF (180 ml) and potassium carbonate (13.6 g, 99 mmol) was added, followed by L-leucine cyclopentyl ester (Intermediate 8) (16 g , 43 mmol). The reaction was stirred for 18h at RT. The residue was diluted with EtOAc (500 ml) and washed with water (3 x 100 ml), dried (Na2SO4), filtered and concentrated to dryness to give the crude product (15.8 g) which was used in the next step without further purification. LCMS purity 60%, m / z 335 [M + H] +.
[0366] The following compounds were prepared in a similar manner:
[0367] Intermediate 1B (S) -2-r (4-Aminobenzh-tert-butoxycarbonylaminol-3-cyclopentyl phenylpropionate
[0371] LCMS purity 75%, m / z 439 [M + H] +.
[0372] Intermediate 1C (S) -r (4-Aminobenzyl) -tert-butoxycarbonylaminol-cyclopentyl phenylacetate
[0376] LCEM purity 100%, m / z 425 [M + H] +.
[0377] Intermediate 1D (S) -2-r (4-Amino-3.5-d¡fluorobenzl) -tert-butoxycarbonylamino1-4-methylcyclopentylpentanoate
[0382] Cyclopentyl 2 (S) - (4-amino-3,5-difluorobenzyl) amino] -4-methylpentanoate (crude 2.54 g, assumed 5.73 mmol) was dissolved in a mixture of THF (25 ml) and water (25 ml). K ² CO ³ (5.15 g, 37.3 mmol) and Boc ² O (8.14 g, 37.2 mmol) were added and stirring was continued at RT for 18 h. The volatiles were removed under reduced pressure and the residual aqueous layer was extracted with EtOAc (50 ml). The organic layer was dried (Na2SO4), filtered, and concentrated under reduced pressure. Purification by flash chromatography (5% EtOAc / heptane) gave the N-Boc protected product (1.0 g, 40%). LCMS purity 89% m / z 441 [M + H] +.
[0384] The benzylamino carbamate used as the starting material was prepared as follows
[0389] To a solution of 4-amino-3,5-difluorobenzaldehyde (0.90 g, 5.73 mmol) in 1/1 of MeOH / DMF (16 ml), L-leucine cyclopentyl ester (Intermediate 8) was added (3.19 g, 8.59 mmol) and K ² CO ³ (1.19 g, 8.59 mmol). The reaction mixture was adjusted to pH 5-6 using glacial acetic acid (dropwise) and stirred for 1 hr before the addition of NaCNBH3 (0.72 g, 11.46 mmol). Stirring was continued at room temperature for 18 h. The reaction mixture was concentrated to remove MeOH, diluted with EtOAc (20 ml), washed with NaHCO3 (5 ml) followed by water (10 ml). The organic layer was dried (Na2SO4), filtered and concentrated in vacuo to give the crude product (2.54 g) which was reacted in the next step without purification. CL purity = 68%.
[0391] The benzaldehyde used as the starting material was prepared in the following manner;
[0396] To a stirred solution of 4-amino-3,5-difluorobenzonitrile (2.0 g, 12.98 mmol) in toluene (16 mL) was added DIBAL (1.5 M in toluene) dropwise at 0 ° C . The reaction mixture was warmed to RT and stirring was continued for 2 h. The reaction was quenched by dropwise addition to aq. Citric acid. 10% (10 ml). EtOAc (50 ml) and aq. Sodium potassium tartrate were added. saturated (Rochelle's salt) (30 ml) and the mixture was vigorously stirred for 20 min. The organic layer was isolated and washed with water (10 ml), dried (Na2SO4), filtered and concentrated to dryness to give a pale yellow solid (1.9 g, 93%). LCMS purity 92%, m / z 158 [M + H] +.
[0398] The following compounds were prepared in a similar manner:
[0399] Intermediate 1E (S) -2-r (4-Am¡no-3.5-d¡fluorobenz¡l) -tert-butoxycarbon¡lam¡no1-3-phenylpropyl cyclopentyl
[0403] LCMS purity 86%, m / z 475 [M + H] +.
[0404] Intermediate 1F (S) -2-r (4-Amino-3.5-d¡fluorobenzl) -tert-butox¡carbon¡lam¡no1-phenylacetate of cyclopentyl
[0408] LCMS purity 86%, m / z 461 [M + H] +.
[0409] Intermediate 2A (S) -rterc-Butox¡carbon¡l - (- 4ir3- (4-fluorophen¡l) -3-oxoprop¡on¡m¡do¡l1am¡nobenz¡l) cyclopentyl aminophenylacetate
[0413] A mixture of 3- (4-fluorophenyl) -3-oxothiopropionimidic acid 4-chlorophenyl ester [WO 03/076405] (300 mg, 0.874 mmol), Intermediate 1C (0.41 g, 0.961 mmol) and acetic acid glacial (3 ml) was stirred at 80 ° C for 2 h. The reaction mixture was evaporated to dryness under reduced pressure to give a thick residue which was triturated with ether (3 ml). The resulting solid was collected by suction filtration. The product was neutralized by partitioning between EtOAc (20 mL) and aq NaHCO3. sat. (10 ml). The organic layer was dried (Na2SO4), filtered, and concentrated in vacuo. Yield = 305 mg (59%). LCMS purity = 75%, m / z 588 [M + H] +. The product was used in the next step without further purification.
[0414] The following starting materials were prepared in an analogous manner:
[0415] ^{Intermediate 2B (S) -2-rferc-Butox¡carbon¡l- (4-ir3- (4-fluorophen¡l) -3-oxoproplon¡m¡do¡} n ^{am¡no} benz¡l) am¡no1- 3-} cyclopentyl phenylpropionate
[0419] From Intermediate 1B, LCEM Purity 76%, m / z 602 [M + H] +.
[0420] ^{Intermediate 2C (S) -2-rterc-Butox¡carbon¡l- (4-ir3- (4-fluorophen¡l) -3-oxoprop¡on¡m¡do¡n am¡no} benz¡nam¡no1- 1-4-} cyclopentyl methylpentanoate
[0424] From Intermediate 1A, LCMS Purity 55%, m / z 568 [M + H] +.
[0425] Intermediate 2D (S) -rterc-Butox¡carbon¡l - (- 4-r3- (2.4-fluorophen¡l) -3-oxoprop¡on¡m¡do¡l1am¡nobenz¡l) cyclopentyl aminophenylacetate
[0429] From Intermediate 1C, LCMS Purity 76%, m / z 606 [M + H] +.
[0430] Intermediate 2E (S) -2-rferc-Butox¡carbon¡l- (4-ir3- (2.4-d¡fluorophen¡l) -3-oxoprop¡on¡m¡doM1am¡no} benz¡l) am¡nol Cyclopentyl -3-phenylpropionate
[0433] From Intermediate 1B, LCMS Purity 78%, m / z 620 [M + H] +.
[0434] Intermediate 2F (S) -2-rterc-Butoxycarbonyl- (4-ir3- (2,4-difluorophenin-3-oxopropionimidoylamino} benzyl) aminol-4-methylpentanoate of cyclopentyl
[0438] From Intermediate 1A, LCMS Purity 76%, m / z 586 [M + H] +.
[0439] Intermediate 2G (S) -rferc-Butoxycarbonyl - (- 4-r3- (3-methyl-4-fluorophenyl) -3-oxopropionimidoylaminobenzyl) aminophenylacetate cyclopentyl
[0443] From Intermediate 1C, LCEM Purity 77%, m / z 602 [M + H] +.
[0444] Intermediate 2H (S) -2-rterc-Butoxycarbonyl- (4-irH3- (3-methyl-4-fluorophenyl) -3-oxopropionimidoylamino} benzyl) -aminol-3-phenylpropionate cyclopentyl
[0448] From Intermediate 1B, LCEM Purity 77%, m / z 616 [M + H] +.
[0449] ^{Intermediate 2I (S) -2-rterc-butoxycarbonyl- (4-ir3- (3-methyl-4-fluorophenyl) -3-oxopropionimidoi} n ^{amino} benzyl) aminol-} 1-4-methylpentanoate cyclopentyl
[0450] From Intermediate 1A, LCMS purity 77%, m / z 582 [M + H] +.
[0451] Intermediate 2J (S) - [tert-Butox¡carbon¡l- (3.5-d¡fluoro-4-l [3- (4-fluorophen¡n-3-oxo-prop¡on¡m¡do¡lalam¡no } benzyl) -aminolphenyl cyclopentyl acetate
[0455] From Intermediate 1F, LCEM purity%, m / z 624 [M + H] +.
[0456] Intermediate 2K (S) -2- [tert-Butox¡carbon¡l- (3.5-d¡fluoro-4-l [3- (4-fluoro-phen¡n-3-oxoprop¡on¡m¡do¡llam No} benzyl) -aminol-4-methylpentanoate cyclopentyl
[0460] From Intermediate 1D. LCEM purity%, m / z 604 [M + H] +.
[0461] Intermediate 2L (S) 2- [tert-Butox¡carbon¡l- (3.5-d¡fluoro-4-l [3- (4-fluoro-phen¡n-3-oxo-prop¡on¡m¡doMlam¡ no} cyclopentyl benzinaminol-3-phenylpropionate
[0465] From Intermediate 1E, LCEM Purity 100%, m / z 638 [M + H] +.
[0466] Intermediate 2M (S) -2- [tert-Butoxycarbonyl- (3.5-d¡fluoro-4-l [3- (2,4-d¡fluorophenyl) -3-oxoprop¡onimidoylamino} benzyl) aminol-4- cyclopentyl methylpentanoate
[0467] From Intermediate 1D, LCEM Purity 86%, m / z 622 [M + H] +.
[0468] Intermediate 3A (SHl4-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n¡l-1-¡l1benc¡l} -tert-butox¡- cyclopentyl carbonylaminophenylacetate
[0472] To a solution of Intermediate 2A (305 mg, 0.52 mmol) in MeOH (5 ml) was added methyl propriolate (70 µl, 0.78 mmol). The mixture was heated at 80 ° C for 3 h. The reaction mixture was concentrated in vacuo and the residue was purified by column chromatography (30% EtOAc / heptane). Yield = 200mg (60%). LCEM purity 80%, m / z 640 [M + H] +.
[0473] The following compounds were produced in a similar way:
[0474] Intermediate 3B (SHl4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n¡l-1-M1benc¡l} -tert-butox¡carbon Lamino) cyclopentyl phenylacetate
[0478] From Intermediate 2D, LCMS Purity 71%, m / z 658 [M + H] +.
[0479] Intermediate 3C S - l4-r6-Am¡no-5- 3-methyl-4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n¡l-1-M1benc¡l} - tertiary
[0480]
[0483]
[0484] Intermediate 3D (S) -2- (l4-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1benzyl} -tert-butox Cyclopentyl ¡-carbonylamino) -3-phenylpropionate
[0488] From Intermediate 2B, CLEM Purity 64%, m / z 654 [M + H] +.
[0489] Intermediate 3E (S) -2 - ({4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-¡l1-benzyl} cyclopentyl tert-butoxycarbonylamino) -3-phenylpropionate
[0493] From Intermediate 2E, LCMS Purity 59%, m / z 672 [M + H] +.
[0494] Intermediate 3F (S) -2- (l4-r6-Am¡no-5- (3-methyl-4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1 -benzl} -tert-butoxycarbonylamino) -3-cyclopentyl phenylpropionate
[0498] From Intermediate 2H, LCMS Purity 87%, m / z 668 [M + H] +.
[0499] Intermediate 3G (S) -2- (ld4-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1benc¡l} -terc- cyclopentyl butoxy-carbonylamino) -4-methylpentanoate
[0503] From Intermediate 2C, LCEM Purity 82%, m / z 620 [M + H] +.
[0504] ^{Intermediate 3I (S) -2- (l4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l-2-oxo-2H-p¡r¡d¡n-1- ¡n benz¡l} cyclopentyl-tert-butoxycarbonylamine)} -4-methylpentanoate
[0508] From Intermediate 2F, LCMS Purity 84%, m / z 638 [M + H] +.
[0509] Intermediate 3J (S) -2- (14-r-Am¡no-5- (3-methyl-4-fluorobenzo¡h-2-oxo-2H-p¡r¡d¡n-1-¡llbenzyl } -te / 'cbutoxycarbonylamino) -4-cyclopentyl methylpentanoate
[0513] From Intermediate 21, LCEM Purity 90%, m / z 634 [M + H] +.
[0514] Intermediate 3K (S) -2- (l4-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡ll-3.5-d¡ fluorobenzyl} -tert-butoxy-carbonylamino) -3-phenyl cyclopentyl propionate
[0518] From Intermediate 2L, LCMS Purity 92%, m / z 690 [M + H] +.
[0519] Intermediate 3L (S) -2- (l4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-Ml-3.5-d Fluorobenzl} -tert-butoxy-carbonylamino) -4-cyclopentyl methylpentanoate
[0523] From Intermediate 2M, LCMS Purity 92%, m / z 674 [M + H] +.
[0524] Intermediate 4A 3- {4-r6-am¡no-5- (4-fluoro-3-methyl-benzo¡l) -2-oxo-2H-p¡r¡d¡n-1-illfenox¡} Propyl ester of methanesulfonic acid
[0528] To a suspension of 6-amino-5- (4-fluoro-3-methyl-benzoyl) -1- [4- (3-hydroxy-propoxy) -phenyl] -1H-pyridin-2-one (100 mg, 0 , 25 mmol) in anhydrous DCM (1 ml) at 0 ° C was added methanesulfonyl chloride (21.5 ml, 0.28 mmol), followed by Et3N (70 µl, 0.50 mmol). The reaction mixture was allowed to warm to RT and stirred for 10-20 min for completion, monitored by TLC (5% MeOH / DCM). The reaction mixture was diluted with DCM (10 ml), washed with 10% citric acid (5 ml), followed by aq NaHCO3. sat. (5 ml) and water (5 ml). The DCM layer was dried (Na2SO4), filtered, and concentrated in vacuo. Yield = 105mg (88%). CLe purity M = 79% m / z = 475 [M + H] +. This material was used in the next step without further purification.
[0529] The alcohol used as the starting material was prepared as follows:
[0530] 6-Amino-5- (4-fluoro-3-methyl-benzoyl) -1- [4- (3-hydroxy-propoxy) -phenyl] -1H-pyridin-2-one was prepared as shown below.
[0531] A mixture of 6-amino-5- (4-fluoro-3-methyl-benzoyl) -1- [4-hydroxy-phenyl] -1H-pyridin-2-one [WO 03/076405] (0.80 g , 2.37 mmol), 3-bromo-1-propanol (0.23 ml, 2.60 mmol), K ² CO ³ (1.37 g, 9.46 mmol), Nal (0.73 g, 4 , 86 mmol) in acetone (20 ml) was heated at 70 ° C for 18 h under an atmosphere of N ² . The reaction mixture was concentrated under reduced pressure, suspended in water (20 ml), and the resulting solid was filtered and washed with ether (0.5 ml). Yield = 0.8g (85%). LCMS purity = 96%, m / z 397 [M + H] +
[0532] The following methanesulfonate intermediates were prepared in a similar manner to Intermediate 4A using methods described in WO 03/076405 for the synthesis of the corresponding 4-hydroxyphenyl intermediates. Intermediate 4B 3l4-r6-Amino-5- (4-fluorobenzoyl) -2-oxo-2H-pyridin-1-illphenoxy} propyl methanesulfonic acid ester
[0536] LCMS purity 66%, m / z 461 [M + H] +.
[0537] Intermediate 4C 3-f4-r6-Amino-5- (2,4-difluorobenzoyl) -2-oxo-2H-pyridin-1-illphenoxy} propyl methanesulfonic acid ester
[0541] LCMS purity 88%, m / z 479 [M + H] +.
[0542] Intermediate 4D 3-l4-r6-Am¡no-5- (4-fluoro-3-methyl-benzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-3.5- d¡fluorophenox¡} prop¡l ester of methanesulfonic acid
[0546] LCMS purity 51%, m / z 511 [M + H] +.
[0547] Intermediate 4E 3-l4-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1-3.5-d¡fluorophenox¡} -prop The ester of methanesulfonic acid
[0551] LCMS purity 72%, m / z 497 [M + H] +.
[0552] Intermediate 4F 3- {4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1-3.5-d¡fluorophenoxy} Propyl ester of methanesulfonic acid
[0556] LCMS purity 81%, m / z 515 [M + H] +.
[0557] The following intermediates were prepared by direct alkylation of 4-hydroxyphenyl intermediates (described in WO03 / 076405) with 1-bromo-5-chloropentane.
[0558] Intermediate 4G 6-Am¡no-1- {4-r (5-chloropent¡nox¡1-2.6-d¡fluorophen¡l} -5- (2.4-d¡fluoro-benzo¡l) p¡r¡d ¡N-2 (1H) -one
[0562] To a solution of 6-amino-5- (2,4-difluorobenzoyl) -1- (2,6-difluoro-4-hydroxyphenyl) -pyridin-2 (1H) -one (300 mg, 0.79 mmol) in acetone (6 ml) under a nitrogen atmosphere were added 1-bromo-5-chloropentane (0.115 ml, 0.87 mmol, 1.1 equiv.), sodium iodide (238 mg, 1.59 mmol, 2 equiv. .) and potassium carbonate (438 mg, 3.17 mmol, 4 equiv.). The mixture was heated at 70 ° C for 16 hours, before being allowed to cool to room temperature and was partitioned between EtOAc (50 ml) and water (50 ml). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. Purification by column chromatography (30% EtOAc in heptane) provided a 3: 2 mixture of the title compound and 6-amino-1- {4 - [(5-iodopentyl) oxy] -2,6-difluorophenyl} - 5- (2,4-Difluorobenzoyl) pyridin-2 (1H) -one (142 mg) which was used without further purification.
[0563] LC / MS: m / z 483, 575 [M + H] +.
[0564] Intermediate 4H 6-Am¡no-1-14-r (5-chloropent¡l) ox¡1-2.6-d¡fluorophen¡l} -5- (4-fluoro-benzo¡l) pyrid¡n- 2 (1H) -one
[0569] To a solution of 6-amino-5- (2,4-fluorobenzoyl) -1- (2,6-difluoro-4-hydroxyphenyl) -pyridin-2 (1H) -one (200 mg, 0.56 mmol) in Anhydrous DMF ⁽⁶ ml) under a nitrogen atmosphere were added 1-bromo-5-chloropentane (0.088 ml, 0.67 mmol, 1.2 equiv.) And potassium carbonate (115 mg, 0.83 mmol, 1, 5 equiv.). The mixture was heated to 40 ° C for 19 hours, before being allowed to cool to room temperature and diluted with EtOAc (20 ml). The solution was washed with water (3 x 20 ml) and brine (20 ml). The organic layer was dried over MgSO ⁴ , filtered, and concentrated under reduced pressure. Purification by column chromatography (20-40% EtOAc in heptane) provided the title compound as a yellow solid (104 mg) which was used without further purification.
[0570] LC / MS: m / z 465 [M + H] +. 1H NMR (300 MHz, CD ³ OD) ⁸ : 7.60 (2H, m), 7.53 (1H, d, J = 9.4 Hz), 7.33 (2H, m), 7.05 (2H, m), 5.72 (1H, d, J = 9.8 Hz), 4.11 (2H, t, J = 6.3 Hz), 3.68 (2H, t, J = 6, 5Hz), 1.84-1.77 (4H, m), 1.56 (2H, m).
[0572] Intermediate 4J 2-l4-r6-Am¡no-5- (4-fluoro-benzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-phen¡l} -et¡l ester of methanesulfonic acid
[0577] To a suspension of 6-Amino-5- (4-fluoro-3-methyl-benzoyl) -1- [4- (2-hydroxy-ethyl) -phenyl] -1H-pyridin-2-one (150 mg, 0 , 43 mmol) in anhydrous DCM (3 ml) at 0 ° C was added methanesulfonyl chloride (34 µl, 0.47 mmol), followed by Et3N (120 ml, 0.85 mmol). The reaction mixture was allowed to warm to RT and stirred for 24 hours to completion. The reaction mixture was diluted with dCm (10 ml), washed with 10% citric acid (5 ml), followed by aq NaHCO3. sat. (5 ml) and water (5 ml). The DCM layer was dried (MgSO4), filtered, and concentrated in vacuo. Yield = 183 mg (crude). LCMS purity = 85% m / z = 431 [M + H] +. This material was used in the next step without further purification. The alcohol used as the starting material was prepared as follows:
[0579] 2- {4- [6-amino-5- (4-fluoro-benzoyl) -2-oxo-2H-pyridin-1-yl] -phenyl} -ethyl acetic acid ester (300 mg) was dissolved in water ( 5 ml) and conc. (5 ml) and heated at 100 ° C for 1 hour. The reaction was then cooled, diluted with 10 ml of water, and filtered. The resulting solid was then dried under reduced pressure to give 264 mg of product, m / z = 353 [M + H] +.
[0581] The 2- {4- [6-amino-5- (4-fluoro-benzoyl) -2-oxo-2H-pyridin-1-yl] -phenyl} -ethyl ester of acetic acid used as the starting material was prepared from as follows:
[0583] A solution of propiolic acid (270 µl, 4.39 mmol) and CDI (712 mg, 4.34 mmol) in THF (13 ml) was warmed from 0 ° C to RT and stirred for 1.5 hours. To this solution was added 2- (4 = {[3- (4-fluoro-phenyl) -3-oxo-propionimidoyl] -amino} -phenyl) -ethyl acetic acid ester (1 g, 2.92 mmol) in THF ⁽⁶ ml) and the reaction was heated to 80 ° C for a maximum period of 2 hours. After cooling and evaporation under reduced pressure, the crude residue was sonicated with methanol (7 ml) before filtration, washing with a minimal amount of methanol. An off white solid was collected (350 mg crude).
[0585] The 2- (4 - {[3- (4-fluoro-phenyl) -3-oxo-propionimidoyl] -amino} -phenyl) -ethyl ester of acetic acid used as starting material was prepared as follows:
[0587] 3- (4-Fluoro-phenyl) -3-oxo-thiopropionimidic acid 4-chloro-phenyl ester (1 g, 2.9 mmol) and 4-aminophenethyl alcohol (418 mg, 3.08 mmol) were dissolved in acid acetic acid (5 ml) and heated at 80 ° C for a period of 24 hours. The reaction was cooled to RT and evaporated under reduced pressure. The crude residue was partitioned between DCM and Na2CO3. The DCM layer was further washed with brine and dried over MgSO4 before evaporation under reduced pressure. The product was isolated (1 g crude) as a 3: 1 mixture of acetylated product: alcohol. This was collected completely unpurified in the cyclization reaction above. Product m / z = 343 [M + H] +, alcohol m / z = 301 [M + H] +.
[0588] Intermediate 56-Am¡no-1-r2.6-d¡fluoro-4- (4-oxo-cyclohex¡lox¡) phen¡l1-5- (4-fluorobenzo¡l) -1H-p¡r D¡n-2-one
[0593] To a solution of 6-amino-1- [4- (1,4-dioxa-spiro [4,5] dec-8-yloxy) -2,6-difluoro-phenyl] -5- (4-fluorobenzoyl) - 1H-pyridin-2-one (0.55 g, 1.10 mmol) in 1,4-dioxane (10 mL) was added aq. 2M (5 ml) at room temperature. Stirring was continued for 18 h. After completion of the reaction, the reaction mixture was diluted with water (10 ml) before evaporation of dioxane under reduced pressure. The residual aqueous solution was extracted with EtOAc (2 x 10 ml). The combined organic layers were dried (Na2SO4), filtered and concentrated to dryness under reduced pressure to give the desired ketone as a white solid (0.43 g, 86%). LCMS purity 98%, m / z 457 [M + H] +, 1H NMR (400 MHz, CDCIa), 8: 2.05-2.15 (2H, m), 2.25-2.45 (4H , m), 2.55-2.70 (2H, m), 4.65-4.75 (1H, m), 5.85 (1H, d), 6.70-6.75 (2H, m ), 7.05-7.15 (2H, m), 7.50-7.65 (3H, m).
[0595] The ketal used as the starting material was prepared as follows:
[0600] To a stirred solution of 1,4-dioxa-spiro [4.5] decan-8-ol (0.5 g, 1.45 mmol) in THF (1.5 ml) was added 6-amino-1- (2,6-Difluoro-4-hydroxyphenyl) -5- (4-fluoro-benzoyl) -1H-pyridin-2-one (prepared by methods described in WO 03/076405) (0.5 g, 1, 39 mmol) and triphenylphosphine (0.38 g, 1.45 mmol) at RT. Diisopropyl azodicarboxylate (0.29 mL, 1.45 mmol) was added dropwise and stirring continued for 18 h. The reaction mixture was evaporated to dryness and purified by column chromatography to provide the desired material as a white solid (0.55 g, 79%). LCMS purity 99%, m / z 501 [M + H] +, 1H NMR (400 MHz, CDCI ³ ), 8: 1.55-1.65 (2H, m), 1.75-2.00 ( 6H, m), 3.85-3.90 (4H, m,), 4.35-4.40 (1H, m), 5.85 (1H, d), 6.10-6.20 ( 2H, m), 7.05-7.15 (2H, m), 7.45-7.60 (3H, m).
[0602] Intermediate 66-Am¡no-5- (4-fluorobenzo¡l) -1- (4-oxo-cyclohex¡l) -1H-p¡r¡d¡n-2-one
[0607] 2M HCl (14 ml) was added to a yellow solution of 6-amino-1- (1,4-dioxa-spiro [4.5] dec-8-yl) -5- (4-fluorobenzoyl) - 1H-pyridin-2-one (664mg, 1.78mmol) in 1,4-dioxane (60ml) at RT. The resulting yellow solution was stirred at RT for 24 h, then diluted with H ² O (30 mL) and concentrated in vacuo to remove 1,4-dioxane, giving a yellow crystalline solid. The solid was isolated by filtration, washed with H ² O, and air dried to give a yellow crystalline solid. Yield = 479mg, 82%. LCMS purity 92%, m / z 329 [M + H] +, 1H NMR (400 MHz, CDCI ³ ), 8: 1.90-2.30 (8H, m), 5.35 (1H, m) , 5.65 (1H, d), 7.05-7.15 (2H, m), 7.30 (1H, d), 7.35-7.45 (2H, m), 11.45 ( 1H, s).
[0609] The pyridone acetal used as the starting material in the above procedure was prepared as follows
[0612] Triethylamine (0.74 ml, 5.31 mmol) was added to a solution of 1- (1,4-dioxa-spiro [4.5] dec-8-yl) -6-ethanesulfinyl-5- (4-fluorobenzoyl ) -1H-pyridin-2-one (1.046 g, 2.42 mmol) in 0.5 M NH ³ in 1,4-dioxane (30 mL) at RT under N ² . The resulting yellow solution was stirred at RT overnight and then concentrated in vacuo to give a yellow solid, which was triturated with TBME, isolated by filtration and washed with TBME to give a pale yellow solid. Yield = 802mg, 89%. LCMS purity 100%, m / z 373 [M + H] +, 1H NMR (400 MHz, CDCI ³ ), 5: 1.90-2.00 (6H, m), 2.60 (2H, m) , 4.15 (4H, m), 5.90 (1H, d), 7.25 (2H, m), 7.55 (1H, d), 7.65 (2H, m).
[0614] The sulfoxide used in the above procedure was prepared as follows
[0619] M-Chloroperbenzoic acid (583 mg, 2.60 mmol) was added in one portion to a yellow solution of 1- (1,4-dioxaspiro [4.5] dec-8-yl) -6-ethylsulfanyl-5 - (4-fluorobenzoyl) -1H-pyridin-2-one (986 mg, 2.36 mmol) in CH ² Cl ² (30 ml) at RT under N ² atmosphere. The resulting yellow solution was stirred at RT overnight and then diluted with CH ² Cl ² (25 mL) and washed with sat. Na2SO3. (2 x 30 mL), sat. NaHCO3 (2 x 30 ml), H ² O (30 ml), dried (Na2SO4), filtered and concentrated in vacuo to give light yellow oil. Yield = 1.046 g, 102%. LCMS 96% purity, m / z 434 [M + H] +.
[0621] The sulfide used in the above procedure was prepared as follows
[0626] 1-Chloro-N, N-2-trimethylpropenylamine (2.03 ml, 15.34 mmol) was added to a colorless solution of propiolic acid (0.94 ml, 15.34 mmol) in anhydrous THF (50 ml) to 0 ° C in an atmosphere of N ² . The resulting colorless solution was stirred at 0 ° C for 2 h, after which time a yellow solution of the acid N- (1,4-dioxa-spiro [4.5] dec-8-yl) -3 was added - (4-fluorophenyl) -3-oxo-thiopropionimidoc (4.667 g, 12.79 mmol) in anhydrous THF (50 ml) for 5 min at 0 ° C. The resulting yellow solution was then allowed to warm to RT and stir for 24 h. The reaction mixture was concentrated in vacuo to give a dark brown oil, which was diluted with EtOAc (20 ml) and allowed to stand at RT overnight to give a crystalline solid which was isolated by filtration and washed with heptane and TBME. Yield = 216 mg. The filtrate was concentrated in vacuo to give a brown solid that was dissolved in CH ² Ch (100 mL) and washed with sat. Na2CO3. (3 x 100 ml), H ² O (2 x 100 ml), dried (Na2SO4), filtered and concentrated in vacuo to give a brown oil. Purification by flash column chromatography (silica, 100% CH ² Cl ² to 30% EtOAc / CH ² Cb) gave the cyclized product after trituration with TBME. Yield = 770 mg. Total yield = 986 mg, 19%. LCEM purity 100%, m / z 418 [M + H] +.
[0628] The thiopropionimidic acid used in the above procedure was prepared as follows:
[0633] K ² CO ³ (16.1 g, 117 mmol) was added to a solution of N- (1,4-dioxaspiro [4.5] dec-8-yl) -3- (4-fluorophenyl) -3-oxothiopropionamide (18.8 g, 55.7 mmol) in acetone (200 ml) at RT / N ² followed by ethyl iodide (6.68 ml, 83.6 mmol). The reaction mixture was stirred at RT / N ² for 2 h and then concentrated in vacuo to give a brown paste which was taken up in EtOAc (300 ml) and washed with H ² O (250 ml). The organic phase was separated and the aqueous phase was extracted with EtOAc (2 x 150 ml). The combined organic phases were dried (Na2SO4), filtered and concentrated in vacuo to give a brown oil. Purification by flash column chromatography (silica, 15% EtOAc / Heptane) gave a yellow oil. Yield = 9.94g, 49%. LCMS purity 94%, m / z 366 [M + H] +.
[0634] The thiopropionamide used in the above procedure was prepared as follows:
[0639] A solution of 4-fluoroacetophenone (6.76 ml, 55.7 mmol) in THF (50 ml) was added slowly over 5 min to a stirred suspension of KOtBu (6.56 g, 58.5 mmol) in THF (40 ml) at 0 ° C. A solution of 8-isothiocyanate-1,4-dioxaspiro [4.5] decane (11.1 g, 55.7 mmol) in THF (30 ml) was added at 0 ° C over 5 min and the resulting mixture was stirred at 0 ° C for 90 min. The reaction mixture was evaporated to dryness, giving a dark brown solid which was used crude in the next step. Yield = 18.8 g, 100%. LCMS purity 55%, m / z 338 [M + H] +.
[0641] The isothiocyanate used in the above procedure was prepared as follows
[0646] Calcium carbonate (13.75 g, 137.4 mmol) was added to a solution of 1,4-dioxaspiro [4.5] dec-8-ylamine (13.5 g, 85.9 mmol) in CH ² Cb (675 ml) and H ² O (330 ml) with vigorous stirring at RT. Thiophosgene (8.5 ml, 111.6 mmol) was added dropwise over 5 min and after the addition was complete, the reaction mixture was stirred at RT for 2 h. The The reaction mixture was diluted with H ² O (600 ml) and extracted into CH ² CI ² (300 ml). The organic phase was dried (Na2SO4), filtered and concentrated in vacuo to give the product. Yield = 8.5g, 50%. LCMS purity 47%, m / z 200 [M + H] +.
[0647] The cyclohexylamine used in the above procedure was prepared as follows:
[0652] 10% Pd (OH) ² / C (1 g) was added to a fine suspension of N, N-dibenzyl-N-1,4-dioxaspiro [4.5] dec-8-ylamine (21.13 g, 62.7 mmol) in EtOH (400 ml) at RT. The resulting mixture was evacuated and purged three times with H ² and then kept under an atmosphere of H ² (balloon) overnight. The reaction mixture was evacuated and purged three times with N ² and then the catalyst was removed by filtration. The filtrate was concentrated in vacuo to give the amine as a colorless oil. Yield = 14.34 g, 99%. LC-MS purity (ELS detection) 100%, m / z 158 [M + H] +.
[0654] The dibenzylamine used in the above procedure was prepared as follows:
[0659] Dibenzylamine (27.8 mL, 145 µmol) was added to a solution of 1,4-dioxaspiro [4.5] decan-8-one (21.5 g, 138 mmol) in DCE (350 mL) at RT at a N ² atmosphere and stirred for 1h. Sodium triacetoxyborohydride (46.7 g, 220 mmol) was added portionwise over 10 min and after the addition was complete, the reaction was stirred at RT / N ² overnight. Saturated NaHCO3 (300 ml) was added, followed by DCM (300 ml) and the reaction mixture was stirred for 30 min. The organic phase was separated and washed with NaHCO3 (300 ml) and brine (300 ml), dried (Na2SO4), filtered and concentrated in vacuo, giving an oil that after trituration with heptane gave a white solid that it was isolated by filtration. Yield = 30.95g, 67%. LCEM purity 100%, m / z 338 [M + H] +.
[0661] Intermediate 7 (S) -2- (tert-Butox¡carbon¡l-13.5-d¡fluoro-4-r3- (4-fluoro-benzo¡l) -6-oxo-1.6-dih¡drop¡r ¡D¡n-2-¡lam¡no1benz¡l} am¡no) -3-phenyl cyclopentyl propionate
[0666] Pyridone was formed as a by-product of the procedure described for the synthesis of Intermediate 3K. LCEM purity 80%, m / z 690 [M + H] +.
[0667] Preparation of amino acid esters (Intermediates 8 to 16)
[0668] Route I. Used for the preparation of Intermediates 8, 9, 13, 14 and 15
[0671] Prepared intermediates:
[0673] Synthesis of compounds indicated in Figure 1
[0674] Route I ( illustrated for Intermediate 9)
[0675] Stage 1 - Ester formation
[0679] To a solution of (S) -2-tert-butoxycarbonylamino-3-cidohexyl-propionic acid (5 g, 19.4 mmol) in DMF (50 ml) at 0 ° C was added cyclopentanol (8.8 ml, 97 , 15 mmol), EDCI (4.09 g, 21.37 mmol) and finally DMAP (237 mg, 1.94 mmol). The reaction mixture was warmed to RT and stirred for 18 h. The DMF was removed in vacuo to give a clear oil. This was separated between water and EtOAc. The organic phase was dried (MgSO4) and concentrated in vacuo. The crude extract was purified by column chromatography (25% EtOAC in heptane) to yield the desired product as a clear oil (14.87 g, 55%). 1H NMR (300MHz, d6-DMsO) 8; 7.09 (1H, d), 5.08 (1H, t), 3.76 (1H, t), 1.50-1.85 (10H, ma), 1.39 (9H, s), 1 0.00-1.25 (9H, ma).
[0680] Step 2- Deprotection of Boc to produce cyclopentyl (2S) -amino (cyclohexyl) acetate hydrochloride (Intermediate)
[0684] The product from Step 1 (14.87 g, 45.69 mmol) was dissolved in DCM (100 ml) and treated with 4M HCl / dioxane (22.8 ml, 91.38 mmol) and the reaction mixture stirred at RT for 24 h. The crude mixture was concentrated under reduced pressure to give an orange oil. This was triturated with Et ² O to give a white precipitate. This was further washed with Et ² O to give the desired product as a white powder (7.78 g, 65%). 1H NMR (300MHz, d6-DMSO) 8; 8.45 (3H, bs), 5.22 (1H, t), 3.28 (1H, d), 1.95-1.50 (10H, ma), 1.30-0.90 ( 9H, Tue).
[0685] Route II ( illustrated for Intermediate 10)
[0686] Step 1 - Ester formation to produce (1S) -2- (cyclopentyloxy) -2-oxo-1-phenylethanaminium 4-methylbenzenesulfonate (Intermediate 10)
[0689] To a suspension of (S) -phenylglycine (5 g, 33.1 mmol) in cyclohexane (150 ml) was added cyclopentanol (29.84 ml, 331 mmol) and p-toluene sulfonic acid (6.92 g, 36 , 4 mmol). The reaction was adjusted with a Dean-Stark receptor and heated to 135 ° C for complete dissolution. After 12 h, the reaction was cooled to RT, leading to precipitation of a white solid. The solid was filtered and washed with EtOAc before drying under reduced pressure to give the required product as a white powder (11.01 g, 85%). 1H NMR (300 µHz, k6-DMSO) 8; 8.82 (2H, bs), 8.73 (1H, bs), 7.47 (7H, m), 7.11 (2H, d), 5.25 (1H, bs), 5.18 ( 1H, m), 2.29 (3H, s), 1.87-1.36 (8H, m).
[0690] Intermediates 11 and 12 were prepared using 2-indanol and α-norborneol, respectively, instead of cyclopentanol (via Route II). In a similar manner, intermediates 13 and 14 were prepared using dimethylaminoethanol and 4- (2-hydroxyethyl) -morpholine, respectively (via Route I). Intermediate 15 was prepared by Route I using commercially available Z-Dab (Boc) -OH (N-a-Z-N-y-Boc-L-2,4-diaminobutyric acid).
[0692] The corresponding (R) -amino acid esters of the above intermediates can be prepared in a similar manner to that shown above, starting from the relevant commercially available (R) -amino acids. Furthermore, the corresponding ferc-butyl esters of Leucine and Phenylglycine are commercially available and used directly where appropriate.
[0694] Examples
[0696] Example 1 (S) -f4-r6-Amino-5- (4-fluorobenzoih-2-oxo-2H-pyridin-1-illbenzylamino} cyclopentyl phenylacetate
[0701] A mixture of Intermediate 3A (80 mg, 0.125 mmol) in a 20% TFA / DCM solution (5 ml) was allowed to stir at RT for 1 h. The reaction mixture was evaporated to dryness and purified by preparative HPLC to give the desired product, yield = 33 mg (40%), LCEM purity = 100% m / z 540 [M + H] +, 1H NMR (400 MHz, DMSO), 8: 1.21-1.82 (8H, m), 4.01-4.14 (2H, m), 5.11-5.21 (2H, m), 5.64 ( 1H, d), 7.21-7.54 (13H, m), 7.62 (1H, d), 10.16 (2H, bs).
[0703] The following examples were prepared in a similar manner to Example 1.
[0705] Example 2 (S) -2- (4- [6-Amino-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1benz¡lamino} - Cyclopentyl 3-phenylpropyl ester
[0710] From Intermediate 3D. LCMS purity 100%, m / z 554 [M + H] +, 1H NMR (400 MHz, DMSO), 8: 1.22-1.83 (8H, m), 3.10 (1H, m), 4.45 (3H, m), 5.19 (1H, m), 5.85 (1H, d), 7.35-7.74 (14H, m), 7.82 (1H, bs), 9 , 96 (1H, bs).
[0711] Example 3 (S) -2-14-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1benz¡lam¡no} - Cyclopentyl 4-methylpentanoate
[0715] From Intermediate 3G. LCMS purity 100%, m / z 520 [M + H] +, 1 H NMR (400 MHz, DMSO) 5: 1.10 ^(6H, m), 1.70 to 2.11 (11H, m) , 4.14-4.53 (3H, m), 5.42 (1H, m, CH), 5.90 (1H, d), 7.49-7.91 (9H, m), 9.83 (2H, sa).
[0716] Example 4 (S) - {4-r6-Amino-5- (3-methyl-4-fluorobenzoyl) -2-oxo-2H-pyridin-1-yl-benzylamino} cyclopentyl phenylacetate
[0720] From Intermediate 3C, LCMS purity 97%, m / z 554 [M + H] +, 1 H NMR (400 MHz, CD ³ OD), 5: 1.30 to 1.81 ⁽⁸ H, m), 2.25 (3H, s), 3.72 (2H, s), 4.34 (1H, s), 5.08 (1H, m), 5.70 (1H, d), 7, 03-7.38 (10H, m), 7.46-7.61 (3H, m).
[0721] Example 5 (S) -2-l4-r6-Am¡no-5- (3-methyl-4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1benc Cyclopentyl lamine} -3-phenylpropyl
[0725] From Intermediate 3F. LCMS purity 100%, m / z 568 [M + H] +, 1 H NMR (400 MHz, DMSO), 5: 1.09 to 1.77 ⁽⁸ H, m), 2.30 (3H, s) , 2.95 (1H, m), 3.14 (2H, s), 4.19-4.42 (3H, m), 5.02 (1H, m), 5.69 (1H, d ), 7.19-7.51 (11H, m), 7.68 (2H, m), 9.79 (2H, bs).
[0726] Example 6 (S) -2-l4-r6-Am¡no-5- (3-MetM-4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1benz¡lam¡ no} -4-cyclopentyl methylpentanoate
[0731] From Intermediate 3J. LCMS purity 100%, m / z 534 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 0.71 (6H, m), 1.32-1.70 (11H, m) , 2.09 (3H, s), 3.72-4.14 (3H, m), 5.03 (1H, m), 5.50 (1H, d), 7.00-7.29 (6H, m), 7.46 (2H, m), 9.40 (2H, bs).
[0733] Example 7 (SH4-r6-Amino-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1benz¡lamino} phenylacetate of c¡ clopent
[0738] From Intermediate 3B. LCMS purity 100%, m / z 558 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 1.33-1.89 (8H, m), 3.71 (2H, m), 4.28 (1H, s), 5.04 (1H, m), 5.61 (1H, d), 6.91 (1H, bs), 7.18-7.60 (13H, m ), 10.05 (1H, bs).
[0740] Example 8 (S) -2- {4-r6-Amino-5- (2,4-difluorobenzoih-2-oxo-2H-pyridin-1-yl-benzylamino} -3-phenylpropionate cyclopentyl
[0745] From Intermediate 3E. LCMS purity 100%, m / z 572 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 1.08-1.76 (8H, m), 2.95 (1H, t), 4.11-4.40 (3H, m), 4.98 (1H, m), 5.68 (1H, d), 6.89 (1H, bs), 7.13-7.50 (12H, m), 7.65 (1H, m), 9.64-10.12 (2H, bs).
[0747] Example 9 (S) -2-14-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1benz¡lam¡no} Cyclopentyl -4-methylpentanoate
[0750] From Intermediate 3I.Pureza LCMS 100%, m / z 538 [M + H] +, 1 H NMR (400 MHz, DMSO) 5: 0.79 ^(6H, m), 1.39 to 1, 78 (11H, m), 3.84-4.22 (3H, m), 5.10 (1H, m), 5.59 (1H, d), 6.79 (1H, bs), 7, 03-7.18 (2H, m), 7.21-7.42 (4H, m), 7.56 (2H, m), 9.54 (1H, bs), 9.92 (1H, sa).
[0752] Example 10 (S) -2-M-r6-Am¡no-5- (2.4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1-3.5-difluorobenz¡lam ¡No} -3-cyclopentyl phenylpropionate
[0757] Starting with Intermediate 3K. LCMS purity 94%, m / z 591 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 1.20-1.90 (10H, m), 3.10 (1H, m), 3.50-3.60 (2H, m), 4.40-4.50 (4H, m), 5.20 (1H, m), 5.90 (1H, d), 7 , 35-7.50 (7H, m), 7.65-7.70 (5H, m), 9.50 (1H, br)
[0759] Example 11 (S) -2-l4-r6-Am¡no-5- (4-fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-¡l1-3.5-d¡fluorobenz¡ lamino} -4-cyclopentyl methylpentanoate
[0764] From Intermediate 3L .Pureza LCMS 96%, m / z 574 [M + H] +, 1 H NMR (400 MHz, CD ³ OD), 5: 0.95 to 1.15 ^(6H, m), 1.65-2.05 (11H, m), 4.15-4.25 (1H, m), 4.35-4.45 (2H, m), 5.35-5.45 (1H, m ), 5.85 (1H, d), 7.10-7.20 (2H, m), 7.45-7.55 (4H, m).
[0766] Example 12 (S) -2- (3-l4-r6-Am¡no-5- (3-methyl-4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1 -M1phenoxy} propylamine) -3-phenyl cyclopentyl propionate
[0771] To a mixture of cyclopentyl ester tosylate salt of L-phenylalanine (Intermediate 16) (218 mg, 0.54 mmol), K ² CO ³ (192 mg, 1.39 mmol), Nal (108 mg, 0.72 mmol) ) a solution of Mesylate Intermediate 4A (170 mg, 0.35 mmol) in THF (2 ml) was added. The reaction mixture was diluted with DMF (2 ml) and heated at 70 ° C for 18 h with stirring. The reaction mixture was cooled to RT, THF was removed by concentration under reduced pressure. The residue was diluted with EtOAc (20 ml) and washed with water (10 ml), dried (Na2SO4), filtered and evaporated to dryness. Preparative HPLC purification provided the desired product, yield = 57 mg, 15%. LCMS purity 97%, m / z 612 [M + H] +, 1 H NMR (400 MHz, CD ³ OD), 5: 1.30-2.00 ⁽⁸ H, m), 2.30 (2H, m), 3.10 (1H, m), 3.40 (1H, m), 4.25 (2H, m), 4.40 (1H, m), 5.20 (1H, m), 5 , 85 (1H, d), 6.90 (2H, m), 7.10 (2H, d), 7.20-7.45 (7H, m), 7.65 (2H, m), 7, 75 (1H, m).
[0773] The following compounds were prepared in a similar manner
[0774] Example 13 (S) - (3-f4-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-3.5-d¡fluorophenox Propylamine) -cyclopentyl phenylacetate
[0779] From Intermediate 4E and cyclopentyl ester tosylate salt of L-phenylglycine (Intermediate 10), LCEM 96% Purity, m / z 620 [M + H] +, 1H NMR (400 MHz, CD ³ OD), ⁸ : 1.40-1.65 (5H, m), 1.80 (2H, m), 1.95 (1H, m), 2.30 (2H, m), 3.15 (1H, m) , 3.30 (1H, m), 4.25 (2H, m), 5.25 (1H, s), 5.40 (1H, m), 5.90 (1H, d), 6.90 (2H, d), 7.30 (2H, t), 7.55-7.60 (5H, m), 7.65 (2H, m) 7.75 (1H, d).
[0781] Example 14 (S) - (3-f4-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-3.5-d¡fluorophenox Propylamine) -cyclopentyl phenylacetate
[0786] From Intermediate 4E and L-phenylalanine cyclopentyl ester tosylate salt (Intermediate 16), LCMS Purity 97%, m / z 634 [M + H] +, 1H NMR (400 MHz, CD ³ OD), ⁸ : 1.30-2.00 ⁽⁸ H, m), 2.30 (2H, m), 3.10 (1H, m), 3.40 (1H, m), 4.25 (2H, m ), 4.40 (1H, m), 5.20 (1H, m), 5.85 (1H, d), 6.90 (2H, m), 7.10 (2H, d), 7 , 20-7.45 (7H, m), 7.65 (2H, m), 7.75 (1H, m).
[0787] Example 15 (S) -2- (3- (4-r6-Am¡no-5- (4-fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-¡l1-3.5- cyclopentyl difluorophenoxyphenox¡} -propylamino) -4-methyl pentanoate
[0792] From Intermediate 4E and L-leucine cyclopentyl ester (Intermediate 8), LCMS Purity 96%, m / z 600 [M + H] +, 1H NMR (400 MHz, CD ³ OD), ⁸ : 1.10 ^(6H, m), 1.70 to 2.0 (12H, m), 2.30 (2H, m), 4.10 (1H, m), 4.25 (2H, m), 4 , 40 (1H, m), 5.40 (1H, m), 5.85 (1H, d), 6.90 (2H, m). 7.10 (2H, d), 7.60. (2H, m), 7.65 (2H, m), 7.75 (1H, m).
[0794] Example 16 N- (3-f4-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -M1-3.5-d¡fluorophenox¡} prop Ethyl nL-leucinate
[0799] From Intermediate 4E and L-leucine ethyl ester, LCMS Purity 98%, m / z 560 [M + H] +, 1H NMR (400 MHz, CD ³ OD), ⁸ : 1.10-1.20 ^(6H, m), 1.45-1.55 (3H, t), 1.65-1.85 (2H, m), 1.90-2.00 (1H, m), 2.15 -2.30 (2H, m), 2.85-3.05 (2H, m), 3.55 (1H, m), 4.35-4.50 (4H, m), 6.00 ( 1H, d), 7.10 (1H, d), 7.45-7.55 (1H, m), 7.80-7.85 (1H, m), 7.95 (1H, d).
[0800] Example 17 (S) - (3-ir6-Am¡no-5- (4-fluoro-3-methylbenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-3.5 -d¡fluorophenox¡} prop¡lam¡no) cyclopentyl phenylacetate
[0804] From Intermediate 4D and cyclopentyl ester tosylate salt of L-phenylglycine (Intermediate 10), LCEM Purity 100%, m / z 634 [M + H] +, 1H NMR (400 MHz, d6-DMSO), ⁸ : 1.20-1.80 ⁽⁸ H, m), 2.0 (2H, m), 2.20 (3H, m), 2.80-3.00 (2H, m), 4.10 (2H , m), 5.10 (1H, m), 5.30 (1H, s), 5.60 (1H, d), 6.95 (1H, d), 7.20 (1H, m), 7 30 (1H, m), 7.40-7.50 ⁽⁸ H, m), 9.65 (1H, m).
[0805] Example 18
[0806] (S) -2- (3-l4-r6-Am¡no-5- (4-fluoro-3-methylbenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1 -3.5-d¡fluorophenox¡} prop¡lam¡no) -3-phen¡lpr cyclopentyl opionate
[0810] From Intermediate 4D and cyclopentyl ester tosylate salt of L-phenylalanine (Intermediate 16), LCEM Purity 97%, m / z 648 [M + H] +, 1H NMR (400 MHz, CD ³ OD), ⁸ : 1.20-1.90 (9H, m), 2.25 (2H, m), 2.35 (3H, s), 3.15 (1H, m), 3.45 (1H, m), 4 , 25 (2H, m), 4.40 (1H, d), 5.20 (2H, m), 5.82 (1H, d), 6.95 (2H, m), 7.20 (1H, m), 7.30-7.50 (7H, m), 7.75 (1H, d). Example 19 (S) -2- (3- {4-r6-Am¡no-5- (4-fluoro-3-methylbenzo¡l) -2-oxo-2H-p¡r¡d¡n- 1-¡l1-3.5-difluorophenox¡} -propylamino) -4-methylpentanoate cyclopentyl
[0814] From Intermediate 4D and L-leucine cyclopentyl ester (Intermediates 8), LCMS Purity 100%, m / z 614 [M + H] +, 1H NMR (400 MHz, d6-DMSO), ⁸ : 0.90 ^(6H, m), 1.60-1.70 (10H, m), 1.90 (2H, m), 2.15 (2H, m), 2.30 (3H, s), 3.00 -3.20 (2H, m), 4.10 (1H, s), 4.20 (2H, m), 5.25 (1H, m), 5.70 (1H, d), 7.05 (1H, d), 7.25 (1H, m), 7.40 (1H, m), 7.50 (1H, m), 7.60 (1H, d).
[0815] Example 20 (S) - (3-l4-r6-Amino-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1-3.5- cyclopentyl difluorophenoxy} propylamine) -phenylacetate
[0820] From Intermediate 4F and cyclopentyl ester tosylate salt of L-phenylglycine (Intermediate 10), CLEM Purity 91%, m / z 638 [M + H] +, 1H NMR (400 MHz, d6-DMSO), ⁸ : 1.50-2.10 ⁽⁸ H, m), 2.30 (2H, m), 3.10-3.25 (2H, m), 4.33 (2H, m), 5.40 (1 H, m), 5.56 (1H, m), 5.90 (1H, d), 7.20 (1H, d) 7.40-7.75 (9H, m), 9.85 (2H, m).
[0822] Example 21 (S) -2- (3-l4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-3.5 -d¡fluorophenox¡} -propylamno) -3-cyclopentyl phenylpropionate
[0827] From Intermediate 4F and cyclopentyl ester tosylate salt of L-phenylalanine (Intermediate 16). LCMS purity 100%, m / z 652 [M + H] +, 1H NMR (400 MHz, d6-DMSO), ⁸ : 1.10-1.80 (9H, m), 2.15 (2H, m), 2.95-3.20 (2H, m), 4.20 (2H, m), 4.40 (1H, m), 5.10 (1H, m), 5.75 (1H, d), 7.06 (2H, d), 7.25-7.58 (9H, m), 9.34 (2H, m).
[0829] Example 22 (S) -2- (3-l4-r6-Amino-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-3.5 -difluorophenoxy} -propylamno) -4-cyclopentyl methylpentanoate
[0834] From Intermediate 4F and L-leucine cyclopentyl ester (Intermediate 8), LCMS Purity 87%, m / z 618 [M + H] +, 1H NMR (400 MHz, d6-DMSO), ⁸ : 1.0 ^(6H, m), 1.75 (9H, m), 1.95 (2H, m), 2.20 (2H, m), 3.10-3.30 (2H, m), 4.16 (1H, m), 4.26 (2H, m), 5.33 (1H, m), 5.80 (1H, d), 7.15 (2H, d), 7.40-7.65 ( 4H, m), 9.13- 9.25 (2H, m).
[0836] Example 23 (SH3-l-r6-Amino-5- (4-fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-M1-phenox¡} propylamino) -fenulacetate cyclopentyl
[0841] From Intermediate 4B and cyclopentyl ester tosylate salt of L-phenylglycine (Intermediate 10), CLEM 95% Purity, m / z 584 [M + H] +, 1H NMR (400 MHz, CD ³ OD), ⁸ : 1.25-1.55 (5H, m), 1.60-1.85 (3H, m), 2.15 (2H, m), 3.00 (1H, m), 3.15 (1 H, m), 4.05 (2H, m), 5.10 (1H, s), 5.25 (1H, m), 5.70 (1H, d), 7.00 (2H, m), 7.15 (4H, m), 7.40-7.50 (7H, m), 7.55 (1H, d).
[0842] Example 24 (S) -2- (3-l4-r6-Amino-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡llfenox¡} prop Cyclopentyl lamine) -3-phenylpropyl
[0847] From Intermediate 4B and L-phenylalanine cyclopentyl ester (Intermediate 16), LCEM Purity 93%, m / z 598 [M + H] +, 1H NMR (400 MHz, CD ³ OD), ⁸ : 1.50 -2.10 ⁽⁸ H, m), 2.50 (2H, m), 3.35 to 3.40 (1H, m), 3.55-3.70 (2H, m), 4.40 -4.50 (2H, m), 4.60 (1H, m), 5.40-5.45 (1H, m), 6.05-6.10 (1H, d), 7, 40-7.65 (11H, m), 7.85 (2H, m). 7.90 (1H, m).
[0848] Example 25 (S) -2- (3- {4-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡llfenox¡} cyclopentyl propylamine) -4-methylpentanoate
[0853] From Intermediate 4B and L-leucine cyclopentyl ester (Intermediate 8), LCMS Purity 97%, m / z 564 [M + H] +, 1H NMR (400 MHz, d6-DMSO), ⁸ : 0.90 ^(6H, m), 1.60-1.75 (10H, m), 1.90 (2H, m), 2.15 (2H, m), 3.10-3.30 (2H, m) , 4.10 (1H, m), 4.15 (2H, m), 5.30 (1H, m), 5.70 (1H, d). 7.15 (2H, d), 7.30 (2H, d), 7.35 (2H, t), 7.50 (1H, d), 7.55 (1H, m) 9.05-30 ( 2H, m).
[0855] Example 26 (S) - (3-14r6-Am¡no-5- (4-fluoro-3-methylbenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l- phenoxy} propylamine) cyclopentyl phenylacetate
[0860] From Intermediate 4A and cyclopentyl ester tosylate salt of L-phenylglycine (Intermediate 10), CLEM 95% Purity, m / z 598 [M + H] +, 1H NMR (400 MHz, d6-DMSO), ⁸ : 1.302.20 (10H, m), 2.30 (3H, m), 2.90-3.10 (2H, m), 4.15 (2H, m), 5.20 (1H, m), 5 , 30 (1H, m), 5.70 (1H, d), 7.10 (2H, d), 7.25-7.40 (5H, m), 7.40-7.50 (3H, m ), 7.55 (5H, m), 9.70 (2H, m).
[0861] Example 27 (S) -2- (3-14-r6-Am¡no-5- (4-fluoro-3-methylbenzo¡l-2-oxo-2H-p¡r¡d¡n-1- ¡Llfenox¡} propylam¡no) -4-cyclopentyl methylpentanoate
[0866] From Intermediate 4A and L-leucine cyclopentyl ester (Intermediate 8), LCEM Purity 89%, m / z 578 [M + H] ⁺ , ¹ H NMR (400 MHz, d ⁶ -DMSO), 8: 0 , 95 (6H, m), 1.55-2.25 (12H, m), 2.30 (3H, m), 2.75-3.30 (2H, m), 4.15 (3H, m ), 5.25 (1H, m), 5.70 (1H, d). 7.15 (2H, d), 7.30-7.40 (4H, m), 7.40-7.50 (2H, m).
[0867] Example 28 (S) - (3-l4-r6-Amino-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-phenox¡} propylamine) cyclopentyl phenylacetate
[0872] From Intermediate 4C and cyclopentyl ester tosylate salt of L-phenylglycine (Intermediate 10), LCEM Purity 99%, m / z 602 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 8: 1.35-2.15 (10H, m), 2.90-3.10 (2H, m), 4.10 (2H, m), 5.25 (1H, m), 5.40 (1H, m), 5.70 (1H, d), 7.10 (2H, d), 7.25-7.30 (4H, m), 7.40-7.50 (2H, m), 7.55 (5H, m), 9.70 (2H, m).
[0874] Example 29 (S) -2- (3-l4-r6-Amino-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1phenox¡} cyclopentyl propylene) -3-phenylpropionate
[0879] From Intermediate 4C and cyclopentyl ester tosylate salt of L-phenylalanine (Intermediate 16), LCMS Purity 99%, m / z 616 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 8: 1.10-1.80 (8H, m), 2.20 (2H, m), 2.95 (1H, m), 3.10-3.30 (2H, m), 3.40 (2H, m), 4.20 (2H, m), 4.40 (1H, m), 5.05 (1H, m), 5.70 (1H, d), 7.15 (2H, d), 7.20-7.55 (11H, m), 9.70 (2H, m).
[0880] Example 30 (S) -2- (3-l4-r6-Amino-5- (2.4d¡fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-¡l1phenox¡} prop Lamino) -4-cyclopentyl methylpentanoate
[0885] From Intermediate 4C and L-leucine cyclopentyl ester (Intermediate 8), LCMS Purity 99%, m / z 582 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 8: 0.95 (6H, m), 1.65-2.15 (13H, m), 3.10-3.20 (2H, m), 4.15 (3H, m), 5.30 (1H, m), 5.70 (1H, d), 7.15 (2H, d), 7.25-7.55 (6H, m), 9.24 (2H, m)
[0887] Example 31 (S) -2- (4-l4-r6-Amino-5- (4-fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-M1-3.5-d¡ Fluorophenoxy} -cyclohexylamino) -4-methylpentanoate cyclopentyl trifluoroacetate
[0892] A suspension of Intermediate 5 (120 mg, 0.263 mmol) and cyclopentyl ester tosylate salt of L-Phenylalanine (Intermediate 16) (98 mg, 0.263 mmol) in MeOH (1.2 ml) was allowed to stir at RT for 1 h before the addition of NaCNBH3 (66 mg, 1.05 mmol). Stirring was continued at RT for 18 h. After the reaction was complete, the reaction mixture was concentrated to dryness and partitioned between EtOAc (10 ml) and water (10 ml). The organic layer was dried (Na2SO4), filtered and concentrated to dryness under reduced pressure and purified by preparative HPLC. This gave the desired product as a TFA salt. Yield = 37 mg (18%).
[0893] LCMS purity 97%, m / z 674 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 5: 1.10-2.20 (16H, m), 2.35-2.45 (1H, m), 2.70-3.00 (2H, m), 3.50-3.55 (1H, m), 4.25-4.35 (1H, m), 4, 95-5.05 (1H, m), 5.70 (1H, d), 6.75 (2H, dd), 7.05-7.25 (7H, m), 7.45-7.55 ( 2H, m), 7.65 (1H, d)
[0895] The following examples were prepared in a similar manner:
[0897] Example 32 (2S) - [(4- {4 [6-amino-5- (4-fluorobenzoyl) -2-oxopyridin-1 (2H) -yl1-3,5-difluorophenoxy) cyclohexyl) amino1- (phenyl) acetate cyclopentyl
[0902] Starting with Intermediate 5 (120 mg, 0.263 mmol) and L-phenylglycine cyclopentyl ester (Intermediate 10). LCMS purity 97%, m / z 660 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 5: 1.30-1.80 (14H, m), 1.90-2.05 (2H, m), 2.35-2.50 (1H, m), 4.35-4.45 (1H, m), 4.50-4.60 (1H, m), 5.05 -5.10 (1H, m), 5.70 (1H, d), 6.75-6.85 (2H, m), 7.10-7.15 (2H, m), 7.20-7 , 35 (5H, m), 7.45-7.55 (2H, m), 7.55-7.60 (1H, m).
[0904] Example 33 (S) -2- (4- {4-r6-Amino-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡ll-3.5 -d¡fluorophenox¡} cyclohex¡lam¡no) -4-cyclopentyl methylpentanoate
[0909] Starting with Intermediate 5 (120 mg, 0.263 mmol) and L-Leucine cyclopentyl ester (Intermediate 8). LCMS purity 100%, m / z 640 [M + H] +, 1 H NMR (400 MHz, CD ³ OD), 5: 0.95-1.05 ^(6H, m), 1.50 to 2, 00 (16H, m), 2.10-2.35 (3H, m), 3.15-3.20 (1H, m), 4.00-4.15 (1H, m), 4, 40 and 4.75 (each 0.5H, m), 5.25-5.35 (1H, m), 5.75 (1H, d), 6.85-6.95 (2H, m ), 7.15-7.25 (2H, m), 7.55-7.65 (2H, m), 7.65-7.70 (1H, m).
[0911] Examples 34 and 35 (S) -2-14-r6-Am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡llclohex¡lam¡no } -3-cyclopentyl propionate
[0916] Intermediate 6 (50 mg, 0.15 mmol) was added to a colorless solution of cyclopentyl ester L-phenylalanine (89 mg, 0.38 mmol) in MeOH (10 mL) at RT / N ² and stirred at RT for 1 h. Glacial AcOH was added dropwise to adjust the pH to ⁶ , followed by NaCNBH3 (38 mg, 0.61 mmol). The resulting colorless solution was stirred at RT overnight and then carefully quenched with sat. NaHCO3. (20 ml) and extracted into CH ² Cl ² (3 x 15 ml). The combined organic phases were washed with 2M HCl (2 x 20 mL) and brine (20 mL), dried (Na2SO4), filtered and concentrated in vacuo to give a cream colored solid. Purification by flash column chromatography (silica, elusion in 40-100% EtOAc / heptane gradient) gave the desired material, separable as two isomers, isomer 1 (Example 34) as a white solid (Yield = 39 mg, 47%) LCEM purity 100 %, m / z 546 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 8: 1.40-2.05 (16H, m), 2.85-3.10 (3H, m ), 3.60-3.70 (1H, m), 4.55-4.65 (1H, m), 5.10-5.20 (1H, m), 5.70 (1H, d), 7.15-7.40 (7H, m), 7.45-7.50 (1H, m), 7.50-7.60 (2H, m) and Isomer 2 (Example 35) LCMS purity 97% , m / z 546 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 8: 1.25-1.85 (12H, m), 1.95-2.20 (2H, m) , 2.45-2.95 (4H, m), 3.00-3.10 (1H, m), 3.65-3.75 (1H, m), 4.55-4.65 (1H, m), 5.05-5.15 (1H, m), 5.65 (1H, d), 7.20-7.35 (7H, m), 7.40-7.50 (1H, m) , 7.50-7.60 (2H, m) as a colorless film (Yield = 15mg, 18%).
[0918] Example 36 (S) - (3-l4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1-3.5- difluorophenoxy} -propylamino) phenyl tere-butyl acetate
[0923] From Intermediate 4F and L-phenylglycine tert-butyl ester, LCMS Purity 93%, m / z 626 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 8: 1.30 (9H , s), 2.15 (2H, m), 3.00-3.15 (2H, m), 4.06 (2H, m), 5.02 (1H, s), 5.70 (1H, d), 6.75 (2H, d), 7.02 (2H, m), 7.30-7.50 (7H, m).
[0925] Example 37 (S) -2- (3-l4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1¡l1-3.5 -difluorophenoxy} -propylamine) -3-phenylpropyl tere-butyl
[0930] From Intermediate 4F and L-phenylalanine tert-butyl ester. LCMS purity 97%, m / z 640 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 8: 1.25 (9H, s), 2.16 (2H, m), 3, 00 (1H, dd), 3.15-3.25 (2H, m), 3.35 (1H, dd), 4.10 (2H, m), 4.20 (1H, m), 5.71 (1H, d), 6.79 (2H, d), 7.02 (2H, t), 7.20-7.30 (5H, m), 2H (2H, m).
[0932] Example 38 (S) -2- (3- {4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l-2-oxo-2H-p¡r¡d¡n-1-¡l1- Tere-butyl 3,5-difluorophenoxy} -propylamino) -4-methylpentanoate
[0937] From Intermediate 4F and L-leucine tert-butyl ester, LCMS Purity 97%, m / z 606 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 8: 0.95 (6H , m), 1.41 (9H, s), 1.61 (1H, m), 1.75 (2H, m), 2.15 (2H, m), 3.22-3.25 (2H, m), 3.88 (1H, m), 4.13 (2H, m), 4.20 (1H, m), 5.40 (1H, s), 5.75 (1H, d), 6 , 85 (2H, d), 7.00 (2H, t), 7.40 (2H, m).
[0938] Example 39 (S) -2- (3-14-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-Mlfenox¡} propylene) -4-methyl tere-butyl pentanoate
[0943] From Intermediate 4C and L-leucine tert-butyl ester, LCMS Purity 91%, m / z 570 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 8: 0.95 (6H , m), 1.45 (9H, s), 1.65 (1H, m), 2.15 (2H, m), 3.15-3.30 (2H, m), 3.85 (1H, m), 4.15 (2H, m), 5.75 (1H, d), 7.00-7.20 (6H, m), 7.35 (2H, m).
[0945] Example 40 (2S) -r (3-l4-r6-Amno-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -ill-3.5-d ¡-Fluorophenoxytpropiham inolffenihacetate of 2.3-d¡h¡dro-1H-¡nden-2-¡lo
[0950] From Intermediate 4F and L-phenylglycine indanyl ester (Intermediate 11), LCMS Purity 96%, m / z 686 [M + H] +, 1H NMR (300 MHz, CD ³ OD), 8: 7.55 -7.47 (2H, m), 7.46-7.31 (5H, m), 7.22-7.10 (6H, m), 6.85 (2H, d, J = 9.6 Hz ), 5.82 (1H, d, J = 9.6Hz), 5.57-5.51 (1H, m), 4.37 (1H, s), 4.13 (1H, t, J = 6.0 Hz), 3.32-3.21 (2H, m), 3.05-2.98 (1H, m), 2.80-2.63 (3H, m), 2.05-1.97 (2H, m).
[0952] Example 41 (2R) -r (3-l4-r6-amino-5- (2.4-d¡fluorobenzo¡l-2-oxop¡r¡d¡n-1 (2H) -Ml-3.5-difluorophenox¡ } -propyl) amnol (phenl) acetate of 2,3-dihydro-1H-inden-2-it
[0957] From Intermediate 4F and D-phenylglycine indanyl ester, LCMS Purity 94%, m / z 686 [M + H] +, 1H NMR (300 MHz, CD ³ OD), 8: 7.56-7.47 (2H, m), 7.38-7.31 (5H, m), 7.28-7.14 (6H, m), 6.85 (2H, d, J = 9.6 Hz), 5, 82 (1H, d, J = 9.6 Hz), 5.57-5.52 (1H, m), 4.37 (1H, s), 4.13 (1H, t, J = 6.0 Hz ), 3.31-3.21 (2H, m), 3.09-2.99 (1H, m), 2.78-2.64 (3H, m), 2.06-1.99 (2H , m).
[0959] Example 42 (2R) -r (3-l4-r6-Amino-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -¡ll-3.5- difluorophenoxy} -propyl) amnol (phenyl) cyclopentyl acetate
[0964] From Intermediate 4F and D-phenylglycine cyclopentyl ester, LCMS Purity 95%, m / z 638 [M + H] +.
[0965] Example 43 (2S) -r (3-l4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l} -2-oxop¡r¡d¡n-1 (2H) -¡l1-3.5- bicyclor2.2.11hept-2-yl difluorophenoxy} propyl) -aminoKphenihacetate
[0969] From Intermediate 4F and L-phenylglycine norborneyl ester (Intermediate 12), LCMS Purity 97%, m / z 664 [M + H] +, 1H NMR (300 MHz, CD ³ OD), ⁸ : 7.50 7.21 (7H, m), 7.02 (2H, t, J = ^{8, 6Hz),} 6.75 (2H, d, J = 9.6Hz), 5.70 (1H, d , J = 9.6Hz), 4.50 (1H, d, J = ^{6, 6Hz),} 4.32 to 4.26 (1H, m), 4.05 (2H, t, J = 5 , 9Hz), 2.71 to 2.56 (2H, m), 2.24 to 1.90 (4H, m), 1.64 to 1.06 ⁽⁸ H, m).
[0970] Example 44 (2R) -r (3-l4-r6-Amino-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -¡l1-3.5- d¡fluorophenox¡} prop¡l) am¡no1- (phenyl) acetate bicyclor2.2.11hept-2-yl
[0974] From Intermediate 4F and D-phenylglycine norborneyl ester, LCEM Purity 98%, m / z 664 [M + H] +, 1H NMR (300 MHz, CD ³ OD), ⁸ : 7.42-7.17 (7H, m), 7.00 (2H, t, J = ^{8, 6Hz),} 6.74 (2H, d, J = 9.1Hz), 5.69 (1H, d, J = 9, 8 Hz), 4.49 (1H, d, J = ^6.8 Hz), 4.32-4.26 (1H, m), 4.03 (2H, t, J = 6.0 Hz), 2 , 70 to 2.52 (2H, m), 2.18 to 1.86 (4H, m), 1.63 to 0.92 ⁽⁸ H, m). Example 45 (S) - (3-l4-r6-Amino-5- (4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-3.5-d Tere-butyl fluorophenoxy} propylamine) -phenylacetate
[0978] From Intermediate 4E and L-phenylglycine tert-butyl ester, LCMS Purity 100%, m / z 608 [M + H] +, 1H NMR (300 MHz, CD ³ OD), ⁸ : 7.55-7 47 (3H, m), 7.30-7.21 ^(6H, m), 7.09 (1H, t, J = 8.7Hz), 6.61 (2H, d, J = 9, 3 Hz), 5.81 (1H, d, J = 9.6 Hz), 4.18 (1H, s), 4.03 (2H, t, J = 6.0 Hz), 2.75-2 , 69 (1H, m), 2.65-2.58 (1H, m), 1.96-1.88 (1H, m), 1.96-1.88 (2H, m); 1.32 (9H, s)
[0979] Example 46 N- (3-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -¡l1-3.5-d¡fluorophenox 2- (dimethylamine) ethyl propil) -L-leucinate
[0983] From Intermediate 4F and Intermediate 13, CLEM Purity 90%, m / z 621 [M + H] +, 1H NMR (300 MHz, DMSO), ⁸ : 10.18 (1H, br s), 9, 50 (1H, bs), 7.57 (1H, c, J = 7.8Hz), 7.39 (2H, m), 7.37-7.15 (3H, m), 7.04 ( 2H, m), 5.73 (1H, d, J = 9.6 Hz), 4.59-4.46 (2H, m), 4.21 (2H, t, J = 9.0 Hz), 4.11 (1H, m), 3.14 (2H, m), 2.86 ^(6H, s), 2.14 (2H, m), 1.74 (2H, m), 0.92 (8H, m).
[0985] Example 47 N- (3-l4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -¡l1-3.5-d¡fluorophenox ¡} Prop¡l) -L-leucinate of 2-morpholin-4-¡let¡lo
[0990] From Intermediate 4F and Intermediate 14, LCEM Purity 90%, m / z 621 [M + H] +, 1H NMR (300 MHz, DMSO), 8: 7.57 (1H, c, J = 7, 5Hz), 7.40 (2H, m), 7.26-7.17 (3H, m), 7.06 (2H, d, J = 10.8Hz), 5.74 (1H, d , J = 9.9 Hz), 4.53 (2H, m), 4.21 (4H, m), 3.80 (4H, m), 3.37 (2H, m), 3.17 (4H , m), 2.15 (2H, m), 1.75 (3H, m), 0.94 (6H, bs).
[0992] Example 48 (2S) -r (3-l4-r6-Amino-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -¡l1-3.5- d¡fluorophenox¡} propyl) -amino1 (cyclohex¡l) cyclopentyl acetate
[0997] From Intermediate 4F and L-cyclohexylglycine cyclopentyl ester (Intermediate 9), CLEM 95% Purity, m / z 664 [M + H] +, 1H NMR (300 MHz, DMSO), 8: 9.15 (1H , bs), 8.95 (1H, bs), 7.62-7.52 (1H, m), 7.46-7.31 (2H, m), 7.27-7.20 (1 H, m), 7.05 (2H, d, J = 10.2 Hz), 5.74 (1H, d, J = 9.6 Hz), 5.30-5.25 (1H, m), 4.20 (2H, t, J = 5.7 Hz), 4.10-3.95 (1H, m), 3.25-2.95 (2H, m), 2.20-2.07 (2H, m), 2.00-1.50 (15H, m), 1.30-1.00 (4H, m), 0.95-0.75 (1H, m)
[0999] Example 49 (2S) -r (3-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -M1-3.5-d OfFluorophenox¡} prop¡l) am¡no1- (cyclohexyl) tere-butyl acetate
[1004] From Intermediate 4F and L-cyclohexylglycine tert-butyl ester, LCMS Purity 95%, m / z 632 [M + H] +, 1H NMR (300 MHz, DMSO), 8: 9.10 (1H, s) ), 8.85 (1H, bs), 7.62-7.52 (1H, m), 7.45-7.31 (2H, m), 7.28-7.20 (1H, m), 7.06 (2H, d, J = 10.2 Hz), 5.74 (1H, d, J = 9.6 Hz), 4.20 (2H, t, J = 5.9 Hz), 3, 95-3.85 (1H, m), 3.25-2.95 (2H, m), 2.20-2.07 (2H, m), 2.00-1.60 (6H, m) , 1.51 (9H, s), 1.35-1.07 (4H, m), 1.00-0.85 (1H, m).
[1006] Example 50 N- (3-l4-r6-Am¡no-5- (2.4-d¡fluorobenzo¡l} -2-oxop¡r¡d¡n-1 (2H) -¡l1-3.5-d¡fluorophenox Propyl) -D-cyclopentyl leucinate
[1009] From Intermediate 4F and D-Leucine Cyclopentyl Ester, LCMS Purity 95%, m / z 618 [M + H] +, 1H NMR (300 MHz, DMSO), 8: 10.10 (1H, sa) , 9.40-9.10 (2H, m), 8.15 (1H, bs), 7.62-7.52 (1H, m), 7.47-7.31 (2H, m), 7.28-7.12 (1H, m), 7.07 (2H, d, J = 10.5 Hz), 5.73 (1H, d, J = 9.6 Hz), 5.30-5 , 20 (1H, m), 4.25-4.00 (3H, m), 3.30-3.00 (2H, m), 2.20-2.00 (2H, m), 1.95 -1.80 (2H, m), 1.75-1.55 (10H, m), 1.00-0.90 (6H, m).
[1011] Example 51 N- (3- {4- [6-amino-5- (2,4-difluorobenzoyl) -2-oxopyridin-1 (2H) -yl] -3,5-difluorophenoxy} propyl) -D-leuccinate ferc-butyl
[1016] From Intermediate 4F and D-Leucine tert-butyl ester, LCMS Purity 95%, m / z 606 [M + H] +, 1H NMR (300 MHz, DMSO), 8: 9.30-9.00 (2H, m), 7.62-7.52 (1H, m), 7.47-7.32 (2H, m), 7.28-7.12 (1H, m), 7.06 ( 2H, d, J = 10.2 Hz), 5.73 (1H, d, J = 9.6 Hz), 4.20 (2H, t, J = 5.7 Hz), 3.99 (1 H, bs), 3.25-2.95 (2H, m), 2.20-2.05 (2H, m), 1.80-1.60 (3H, m), 1.49 (9H, s), 0.95 (6H, d, J = 4.8 Hz).
[1018] Example 52 (2S) -4-Amino-2-r (3-14-r6-amino-5- (2.4-d¡fluorobenzo¡n-2-oxop¡r¡d¡n-1 (2H ) -M1-3.5-d¡fluorophenoxy} prop¡l) amino1butanoate cyclopentyl
[1023] From Intermediate 4F and Intermediate 15, CLEM Purity 90%, m / z 605 [M + H] +, 1H NMR (300 MHz, CD ³ OD), ⁸ : 7.46 - 7.55 (2H, m), 7.12 (2H, t, J = 8.7 Hz), 6.93 (2H, d, J = 9.6 Hz), 5.81 (1H, d, J = 9.6 Hz) , 5.37-5.44 (1H, m), 4.20-4.31 (4H, m), 3.33-3.42 (1H, m), 2.25-2.49 (4H , m), 1.91-2.08 (2H, m), 1.65-1.89 (7H, m).
[1025] Example 53 M- (5-f4-r6-am¡no-5- (2.4-d¡fluorobenzo¡n-2-oxop¡r¡d¡n-1 (2H) -¡l1-3.5-d¡fluorophenox¡ } pentyl) -L-cyclopentyl leucinate
[1030] From Intermediate________ ⁴ ®. To a solution of 6-amino-1- {4 - [(5-chloropentyl) oxy] -2,6-difluorophenyl} -5- (2,4-difluorobenzoyl) pyridin-2 (1H) -one (138 mg, 0.29 mmol) in anhydrous DMF (3 ml) under a nitrogen atmosphere, cyclopentyl L-leucinate (Intermediate 8) (284 mg, 1.43 mmol, 5 equiv.), Sodium iodide (86 mg , 0.57 mmol, 2 equiv.) And N, N-diisopropylethylamine (0.052 ml, 0.29 mmol, 1 equiv.). The mixture was heated at 90 ° C for 16 hours, before being allowed to cool to room temperature and was diluted with EtOAc (25 ml). The solution was washed with water (2 x 25 ml) and brine (25 ml). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. Purification by column chromatography (3-4% MeOH in DCM), followed by preparative HPLC provided the title compound as a colored solid (96 mg, 52% yield).
[1031] LC / MS: m / z 646 [M + H] +. 1H NMR (300 MHz, CD ³ OD) 8: 7.56 to 7.47 (2H, m), 7.13 (2H, m), 6.88 (2H, m), 5.82 (1H, d, J = 9.8 Hz), 5.23 (1 H, t, J = 4.1 Hz), 4.11 (2H, t, J = 6.3 Hz), 3.28 (1 H, m), 2.61-2.51 (2H, m), 1.95-1.41, (17H, ma), 0.98-0.93 (6H, m).
[1032] Example 54 N- (5-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -¡l1-3.5-d¡fluorophenoxy } pent¡l) -L-tere-butyl leucinate
[1037] From Intermediate________4G. To a solution of 6-amino-1- {4 - [(5-chloropentyl) oxy] -2,6-difluorophenyl} -5- (2,4-difluorobenzoyl) pyridin-2 (1H) -one (96 mg, 0.20 mmol) in anhydrous DMF (3 ml) under a nitrogen atmosphere was added tert-butyl L-leucinate hydrochloride (198 mg, 0.99 mmol, 5 equiv.), Sodium iodide (60 mg, 0.40 mmol, 2 equiv.) And N, N-diisopropylethylamine (0.072 ml, 0.40 mmol, 2 equiv.). The mixture was heated at 90 ° C for 20 hours, before being allowed to cool to room temperature and diluted with EtOAc (20 ml). The solution was washed with water (2 x 20 ml) and brine (20 ml). The organic layer was dried over MgSO ⁴ , filtered, and concentrated under reduced pressure. Purification by column chromatography (1-3% MeOH in DCM), followed by preparative HPLC provided the title compound as a white solid (23 mg, 18% yield).
[1038] LC / MS: m / z 634 [M + H] +. 1H NMR (300 MHz, CD ³ OD) ^8: 7.56 to 7.47 (2H, m), 7.14 (2H, m), 6.89 (2H, m), 5.81 (1H, d, J = 9.6 Hz), 4.11 (2H, t, J = 6.2 Hz), 3.19 (1H, m), 2.60-2.53 (2H, m), 1 , 89 to 1.84 (2H, m), 1.72 to 1.41 (16H, m), 0.96 ^(6H, m).
[1039] Example 55 N- (5-14-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -M1-3.5-d¡fluorophenox¡} pent ¡NL-cyclopentine leucine
[1044] From Intermediate 4H. To a solution of 6-amino-1- {4 - [(5-chloropentyl) oxy] -2,6-difluorophenyl} -5- (4-fluorobenzoyl) pyridin-2 (1H) -one (99 mg, 0, 21 mmol) in anhydrous DMF (3 ml) under nitrogen atmosphere was added cyclopentyl L-leucinate (Intermediate 8) (212 mg, 1.06 mmol, 5 equiv.), Sodium iodide (64 mg, 0, 43 mmol, 2 equiv.) And W, W-diisopropylethylamine (0.039 ml, 0.21 mmol, 1 equiv.). The mixture was heated to 90 ° C for 20 hours, before being allowed to cool to room temperature and was diluted with EtOAc (25 ml). The solution was washed with water (2 x 25 ml) and brine (25 ml). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. Purification by column chromatography (2% MeOH in DCM), followed by preparative HPLC provided the title compound as a yellow solid (64 mg, 48% yield).
[1045] LC / MS: m / z 628 [M + H] +. 1H NMR (300 MHz, CD ³ OD) ⁸ : 7.71 (1H, d, J = 9.6 Hz), 7.62 (2H, m), 7.26 (2H, m), 6.89 ( 2H, m), 5.81 (1H, d, J = 9.6Hz), 5.23 (1H, t, J = 5.3Hz), 4.11 (2H, t, J = 6 , 4 Hz), 3.28 (1H, m), 2.55 (2H, m), 1.91 to 1.48 (17H, m), 0.98 to 0.93 ^(6H, m).
[1047] Example 56 N- (14-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -¡l1phenoxy} acetyl) -L-leuc¡ cyclopentyl nato
[1052] LC / MS: m / z 564 [M + H] +. 1H NMR (300 MHz, DMSO-d6) ⁸ : 8.47 (1H, d, J = 7.7 Hz), 7.56 (2H, m), 7.45 (1H, d, J = 9, 6Hz), 7.38-7.24 (4H, m), 7.15 (2H, m), 5.69 (1H, d, J = 9.8Hz), 5.09 (1H, t, J = 5.3 Hz), 4.63 (2H, m), 4.31 (1H, m), 1.84-1.79 (2H, m), 1.66-1.53 ( 9H, m), 0.92 to 0.86 ^(6H, m).
[1053] To a solution of 6-amino-5- (4-fluorobenzoyl) -1- (4-hydroxyphenyl) pyridin-2 (1H) -one [WO 03/076405] (100 mg, 0.31 mmol) in anhydrous DMF (3 ml) under a nitrogen atmosphere were added cyclopentyl N- (bromoacetyl) -L-leucinate (109 ml, 0.34 mmol, 1.1 equiv.) And potassium carbonate (51 mg, 0.37 mmol, 1.2 equiv.). The mixture was heated at 40 ° C for 16 hours, before being allowed to cool to room temperature it was added to water (20 ml). The mixture was extracted with EtOAc (3 x 15 ml) and the combined extracts were washed with water (2 x 40 ml) and brine (40 ml). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. Purification by column chromatography (2-3% MeOH in DCM), followed by trituration with minimal MeOH provided the title compound as a white solid (91 mg, 52% yield).
[1055] Cyclopentyl N- (bromoacetyl) -L-leucinate was synthesized from cyclopentyl L-leucinate in one step, the details of which are given below.
[1060] To a solution of cyclopentyl L-leuccinate (Intermediate 8) (568 mg, 2.84 mmol) in DCM (6 ml) was added triethylamine (0.24 ml, 2.84 mmol, 1 equiv.) Dropwise. and bromoacetyl chloride (1.44 ml, 3.13 mmol, 1.1 equiv.). The mixture was stirred at room temperature for 20 hours, diluted with DCM (50 ml) and washed with water (50 ml) and brine (50 ml). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure to provide a crude mixture containing the title compound (902 mg) that was used without further purification. LC / MS: m / z 320/322 [M + H] +.
[1062] ^{Example 57 N-r2- (4-16-am¡no-5-r (4-fluorophenyl) carbon¡} n ^{-2-oxop¡r¡d¡n-1 (2H) -¡l} phen¡l ) cyclopentyl ethyl1-L-leucinate}
[1067] From Intermediate 4J and L-Leucine cyclopentyl ester (Intermediate 8). LC / MS: m / z 534 [M + H] +. 1H NMR (300 MHz, DMSO-d6) 8: 8.47 (1H, d, J = 7.7Hz), 7.56 (2H, m), 7.45 (1H, d, J = 9 , 6Hz), 7.38-7.24 (4H, m), 7.15 (2H, m), 5.69 (1H, d, J = 9.8Hz), 5.09 (1H , t, J = 5.3 Hz), 4.63 (2H, m), 4.31 (1H, m), 1.84-1.79 (2H, m), 1.66-1.53 (9H, m), 0.92-0.86 (6H, m).
[1069] The following examples were synthesized in a similar way:
[1071] Example 58 N-r2- (4-16-amino-5-r (4-fluorophenyl) carbonM1-2-oxop¡r¡d¡n-1 (2H) -¡l} phenl) etM1 -L-tere-butyl leucinate
[1076] From Intermediate 4J and t-butyl ester of L-Leucine. LC / MS: m / z 522 [M + H] +. 1H NMR (300 MHz, DMSO-d6) 8: 9.40-9.10 (2H, m), 7.59-7.44 (5H, m), 7.38-7.30 (4H, m) , 5.71 (1H, d, J = 9.6Hz), 4.00 (1H, bs), 3.40-3.28 (1H, m), 3.25-3.15 ( 1H, m), 3.10-3.00 (2H, m), 1.80-1.70 (3H, m), 0.96 (6H, d, J = 5.1 Hz).
[1077] Example 59 N-r2- (4-16-am¡no-5-r (2.4-d¡fluorophen¡l) carbon¡l1-2-oxop¡r¡d¡n-1 (2H) -¡l} fen ¡L) eth¡l1-L-cyclopentyl leucinate
[1082] Example 59 was synthesized by a similar route to that of Example 57 using 3- (2,4-difluoro-phenyl) -3-oxo-thiopropionimide 4-chloro-phenyl ester. LC / MS: m / z 552 [M + H] +. 1H NMR (300MHz, DMSO-da) 8: 10.07 (1H, bs), 9.35 (2H, bs), 7.55-6.95 (8H, m), 5.72 (1H, d, J = 9.9 Hz), 5.27 (1H, t, J = 5.7 Hz), 4.15-4.00 (1H, m), 3.41-3.15 (2H, m ), 3.10-3.00 (2H, m), 1.96-1.80 (2H, m), 1.78-1.55 (9H, m), 0.95 (6H, d, J = 5.1 Hz).
[1084] Example 60 N-r2- (4-16-am¡no-5-r (2.4-d¡fluorophen¡l) carbon¡l1-2-oxop¡r¡d¡n-1 (2H) -¡l} fen ¡L) tert-butyl ethyl1-L-leucinate
[1089] Example 60 was synthesized by a route similar to Example 57 using 3- (2,4-difluorophenyl) -3-oxo-thiopropionimide 4-chloro-phenyl ester. LC / MS: m / z 540 [M + H] +. 1H NMR (300MHz, DMSO-d6) 8: 10.07 (1H, bs), 9.30 (2H, bs), 7.55-6.94 (8H, m), 5.72 (1H , d, J = 9.6 Hz), 4.05-3.93 (1H, m), 3.40-3.10 (2H, m), 3.08-3.00 (2H, m) , 1.80-1.65 (3H, m), 1.50 (9H, s), 0.96 (6H, d, J = 5.1 Hz).
[1091] Example 61 (2SHr2- (4-16-Amino-5-r (4-fluorophenyl) carbonM1-2-oxop¡r¡d¡n-1 (2H) -¡l} phen¡l) et¡ l1am¡no} (phenyl) cyclopentyl ethanoate
[1096] From Intermediate 4J and cyclopentyl ester of L-Phenylglycine (Intermediate 10). LC / MS: m / z 554 [M + H] +, 1H NMR (300 MHz, CDCIa) 8: 10.35 (1H, bs), 7.60-7.13 (14H, m), 5, 91 (1H, d, J = 10.2 Hz), 5.22-5.14 (1H, m), 4.36 (1H, s), 3.00-2.85 (4H, m), 2 , 16 (1H, bs), 1.99-1.43 (8H, m).
[1098] Example 62 (2SHr2- (4-16-Amino-5-r (4-fluorophen¡hcarbonM1-2-oxop¡r¡d¡n-1 (2H) -¡l} phenenet¡l1am¡no} tert-butyl (phenyl) ethanoate
[1103] From Intermediate 4J and t-butyl ester of L-Phenylglycine. LC / MS: m / z 542 [M + H] + 1H NMR (300 MHz, CDCI ³ ) 8: 10.30 (1H, bs), 7.51-7.45 (3H, m), 7.38 (2H, d, J = 6.9Hz), 7.27-7.04 (9H, m), 5.82 (1H, d, J = 9.6Hz), 4.20 (1H, s), 2.86-2.75 (4H, m), 2.04 (1H, bs), 1.31 (9H, s).
[1104] Example 63 N-r2- (4- {6-am¡no-5-r (4-methylphenyl) carbon¡ll-2-oxop¡r¡d¡n-1 (2H) -¡l} phenyl) ethyl-L- cyclopentyl leuccinate
[1109] Example 63 was prepared by a methodology similar to Example 59 using 3- (4-methylphenyl) -3-oxo-thiopropionimidic acid 4-chloro-phenyl ester, prepared by a method similar to that used for Intermediate 4J. LC / MS: m / z 530 [M + H] +. 1H NMR (300 MHz, CDCIa) 8: 7.65 (1H, d, J = 9.9Hz), 7.47 (4H, m), 7.26 (4H, m), 5.89 (1 H, d, J = 9.9Hz), 5.20 (1H, m), 3.25 (1H, t, J = 7.2Hz), 2.87 (4H, m), 2.44 (3H, s), 1.99-1.53 (9H, m), 1.42 (2H, t, J = 6.3Hz), 0.91 (6H, m).
[1111] Example 64 M-r2- (4- {6-am¡no-5-r (4-methoxyphen¡l) carbon¡ll-2-oxop¡r¡d¡n-1 (2H) -¡l} phenyl) ethyl-L-cyclopentyl leucinate
[1116] Example 64 was prepared by a methodology similar to Example 59 using 3- (4-methoxy-phenyl) -3-oxo-thiopropionimide 4-chloro-phenyl ester, prepared by a method similar to that used for Intermediate 4J. LC / MS: m / z 546 [M + H] +. 1H NMR (300 MHz, CDCIa) 8: 7.67 (1H, d, J = 9.9Hz), 7.55 (2H, d), 7.46 (2H, d), 7.24 (2H , d, J = 8.4 Hz), 6.98 (2H, d, J = 6.9 Hz), 5.90 (1H, d, J = 9.6 Hz), 5.18 (1H , m), 3.88 (3H, s), 3.24 (1H, t, J = 7.2 Hz), 2.87 (4H, m), 1.97-1.53 (9H, m ), 1.43 (2H, t), 0.90 (6H, m).
[1118] Example 65 N-r2- (4-16-am¡no-5-r (4-chlorophenyl) carbon¡ll-2-oxop¡r¡d¡n-1 (2H) -¡l} phen¡l ) ethinL-cyclopentyl leucinate
[1123] Example 65 was prepared by a methodology similar to Example 59 using 3- (4-chloro-phenyl) -3-oxo-thiopropionimidic acid 4-chloro-phenyl ester, prepared by a method similar to that used for Intermediate 4J. LC / MS: m / z 551 [M + H] +. 1H NMR (300 MHz, CDCI ³ ) 8: 7.40 (7H, m), 7.16 (2H, d, J = 8.4 Hz), 5.82 (1H, d, J = 9.9 Hz ), 5.11 (1H, m), 3.17 (1H, t, J = 7.5 Hz), 2.78 (4H, m), 1.92-1.43 (9H, m), 1 , 35 (2H, t), 0.82 (6H, dd).
[1125] ^{Example 66 N-r3- (4-16-amino-5-r (4-fluorophenyl) carbon¡} n ^{-2-oxop¡r¡d¡n-1 (2H) -¡l} -3- fluorophenox¡) prop¡n -L-} cyclopentyl ^leucinate
[1130] Example 66 was prepared by a methodology similar to Example 25 using 6-amino-5- (4-fluoro-3-methyl-benzoyl) -1- [2-fluoro-4-hydroxy-phenyl] -1H-pyridine -2-one [WO 03/076405]. Purity LCMS 97%, m / z 582 [M + H] +, 1H NMR (400 MHz, da-DMSO), 5: 7.57 ( 2H, m), 7.48 (1H, d, J = 9, 6Hz), 7.34 (3H, m), 7.10 (1H, dd, J = 11.9, 2.3Hz), 7.00 (1H, dd, J = 9.7, 2.3 Hz), 5.70 (1H, d, J = 9.6Hz), 5.12 (1H, m), 4.11 (2H, t, J = 6.2Hz), 3.14 ( 1H, m), 2.68 (1H, m), 1.98 (1H, m), 1.88-1.82 (4H, m), 1.67-1.57 (7H, m ), 0.88 ^(6H, t, J = 7.2Hz).
[1132] ^{Example 67 N-r3- (4-16-amino-5-r (4-fluorophenyl) carbon¡} n ^{-2-oxop¡r¡d¡n-1 (2H) -¡l} phenM) propM1} Cyclopentyl ^-L-leucinate
[1137] Example 61 was synthesized by a similar route to Example 57 using 3- (4-amino-phenyl) -propan-1-ol. LC / MS: m / z 548 [M + H] +. 1H NMR (300MHz, DMSO-d6) 5: 9.13 (2H, bs), 7.59-7.52 (2H, m), 7.50-7.42 (3H, m), 7.39 -7.25 (4H, m), 5.70 (1H, d, J = 9.6Hz), 5.28-5.24 (1H, m), 4.04 (1H, bs), 3.35-2.85 (2H, m), 2.80-2.70 (2H, m), 2.10-1.80 (4H, m), 1.75-1.55 (10H, m ), 0.93 ^(6H, d, J = 3.4 Hz).
[1139] 3- (4-Amino-phenyl) -propan-1-ol was synthesized in a one-step procedure from 4-nitro cinnamyl alcohol as follows:
[1141] To a solution of 4-nitro cinnamyl alcohol (2 g, 11.1 mmol) in methanol (30 ml) under a nitrogen atmosphere was added Raney nickel (2 ml suspension in water). The reaction was then exposed to hydrogen gas and stirred under an atmosphere of hydrogen for 12 hours to complete the reaction. The reaction mixture was filtered through Celite, washing with methanol and ethyl acetate. The filtrate was then concentrated under reduced pressure before purification by column chromatography (8: 2 EtOAc: Hexane) to give the required product (1.68 g, 95%) as a yellow solid.
[1143] Example 68 N2-r3- (4-16-am¡no-5-r (2.4-d¡fluorophen¡l) carbon¡l1-2-oxop¡r¡d¡n-1 (2H) -¡l} - 3.5-d¡fluorophenox¡) propM1-L¡s¡s¡nate de cyclopent¡lo
[1148] Example 68 was synthesized by a similar route to Example 52 using (Z) -cyclopentyl ester of L-Lysine. LC / MS: m / z 633 [M + H] +. 1H NMR (300 MHz, CD ³ OD) 5: 7.45 to 7.54 (2H, m), 7.12 (2H, t, J = ^{8, 6Hz),} 6.93 (2H, d, J = 9.8 Hz), 5.81 (1H, d, J = 9.8 Hz), 5.33-5.40 (1H, m), 4.25 (2H, t, J = 5.1 Hz), 4.10-4.16 (1H, m), 2.96 (2H, t), 2.27-2.35 (2H, m), 1.63-2.12 (16H, m ).
[1150] Example 69 M2-r3- (4-16-am¡no-5-r (2.4-d¡fluorophen¡l) carbon¡l1-2-oxop¡r¡d¡n-1 (2H) -¡l} - 3.5-d¡fluorophenox¡) propM1-L¡s¡nate of tert-feut¡nate
[1155] Example 69 was synthesized by a route similar to that of Example 52 using (Z) -tbutyl ester of L-Lysine. LC / MS: m / z 621 [M + H] +, 1H NMR (300 MHz, CD ³ OD) 5: 7.48 (2H, dd, J = 9.7, 2.7 Hz), 7.12 (2H, t, J = ^8.6 Hz), ^{6.8 8} (2H, d, J = 9.2 Hz), 5.81 (1H, d, J = 9.6 Hz), 4.18 ( 2H, t, J = 6.2Hz), 3.15 (H, t, J = ^{6, 6Hz),} 2.76 to 2.87 (3H, m), 2.68 (1H, dt, J = 11.5, 6.9 Hz), 1.97-2.05 (4H, m), 1.64 (4H, dt, J = 6.1 Hz), 1.01 (9H, s).
[1156] Example 70 N-r2- (3-16-amino-5-r (4-fluorophenyl) carbon¡l-2-oxop¡r¡d¡n-1 (2H) -¡l} phen¡l ) cyclopentyl ethyl1-L-leucinate
[1161] Example 70 was synthesized using a methodology similar to Intermediate 4J (using 3-aminophenethyl alcohol instead) and L-Leucine cyclopentyl ester (Intermediate 8).
[1162] LC / MS: m / z 534 [M + H] +. 1 H NMR (300 MHz, DMSO-da) 8: 9.40-9.00 (2H, m), 7.65-7.44 (5H, m), 7.38-7.11 (4H, m) , 5.72 (1H, d, J = 9.9Hz), 5.30-5.20 (1H, m), 4.10-4.00 (1H, m), 3.45-3 , 15 (2H, m), 3.10-3.00 (2H, m), 1.95-1.80 (2H, m), 1.75-1.55 (9H, m), 9.93 (6H, d, J = 4.8 Hz).
[1164] Example 71 (S) -2-13.5-D¡fluoro-4-r3- (4-fluorobenzo¡l) -6-oxo-1.6-d¡h¡drop¡r¡d¡n-2-¡lamino1benc Lamino} -3-cyclopentyl phenylpropionate
[1169] A solution of Intermediate 7 (20 mg) in 20% TFA / DCM (0.5 ml) was allowed to stand at RT for 1 hr. After completion, the reaction mixture was blown evaporated to dryness under a moderate stream of N ² . DCM (0.5 ml) was added and blown under N ² atmosphere. Drying in a N ² atmosphere was continued overnight. Yield = 20mg, 98%.
[1170] LCMS 96% purity, m / z 590 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 8: 0.75-1.33 (8H, m), 2.90 (1H, m ), 3.50 (2H, m), 4.25 (3H, m), 4.93 (1H, m), 5.70, (1H, m), 5.98 (1H, m), 7, 15-7.62 (11H, m), 9.7 (1H, bs), 10.42 (0.5H, bs)
[1171] Reference Example 72 (SH4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1benzylamine} -phenylacetic acid
[1176] To a solution of Example 7 (100mg, 0.179mmol) in THF (1ml) and MeOH (0.5ml) was added 2M NaOH (aq, 1ml). The mixture was allowed to stir at RT for 3h, evaporated to near dryness, acidified using dropwise addition of 1M HCl and extracted with EtOAc (5 ml). The EtOAc layer was concentrated in vacuo to give the crude oil. LCMS shows 80% product m / z = 490 [M + H] + and 20% impurity m / z 470 [M + H] +. Preparative HPLC purification provided the desired product. Yield = 34mg, 31%). LCMS purity 100%, m / z 490 [M + H] +, 1H NMR (400 MHz, DMSO), 8: 3.64 (2H, m, CH ² ), 4.06 (1H, s, CH) , 5.50 (1H, d, Ar), 6.75 (1H, bs, NH), 6.96 (13H, m, Ar), 9.84 (1H, brs, NH).
[1178] The following compounds were prepared in a similar manner:
[1179] Reference Example 73 Acid (S) -2-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1benz¡lam No} -3-phenylpropionic
[1184] From Example 8. LCMS purity 99%, m / z 504 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 2.96-3.09 (3H, m), 3.81 (1H, d), 3.98 (1H, d), 5.76 (1H, d), 7.00 (1H, bs), 7.22-7.40 (9H, m), 7.41- 7.61 (4H, m), 10.11 (1H, bs).
[1186] Reference Example 74 Acid (S) -2-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1benz¡lam No! -4-methylpentanoic
[1191] From Example 9. LCMS purity 91%, m / z 470 [M + H] +, 1H NOR (400 MHz, DMSO), 5: 0.71 (6H, m), 1.40 (2H, m ), 1.60 (1H, m), 3.44 (1H, m), 3.89 (2H, s), 5.49 (1H, d), 6.70 (1H, bs), 6.94-7.08 (2H, m), 7.14-7.33 (4H, m), 7.46 (2H, m), 9.86 (1H, bs).
[1193] Reference Example 75 (SH4-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1benzylamine} -phenylacetic acid
[1198] From Example 1. LCMS purity 100%, m / z 472 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 4.13 (1H, d), 4.22 (1H, d ), 5.18 (1H, s), 5.73 (1H, d), 7.30-7.61 (13H, m), 7.73 (1H, d), 10.09 (2H, sa) .
[1200] Reference Example 76 (SH4-r6-am¡no-5- (3-methyl-4-fluorobenzo¡n-2-oxo-2H-pyr¡din-1-M1-benzylamino} phenylacetic acid
[1201] From Example 4. LCMS purity 86%, m / z 486 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 2, 20 (3H, s), 3.98 (1H, d ), 4.06 (1H, d), 5.07 (1H, s), 5.62 (1H, d), 7.12-7.50 (12H, m), 7.61 (1H, d ), 9.90 (2H, bs).
[1202] Reference Example 77 Acid (S) -2-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1benz¡lam No! -4-methylpentanoic
[1206] From Example 9. LCMS purity 91%, m / z 470 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 0.71 (6H, m), 1.40 (2H, m ), 1.60 (1H, m), 3.44 (1H, m), 3.89 (2H, s), 5.49 (1H, d), 6.70 (1H, bs), 6.94 -7.08 (2H, m), 7.14-7.33 (4H, m), 7.46 (2H, m), 9.86 (1H, bs).
[1207] Reference Example 78 Acid (S) -2-l4-r6-am¡no-5- (4-fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-¡l1benzylam¡no} - 3-phenylpropropionic
[1211] From Example 2. LCMS purity 100%, m / z 486 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 3.00 (2H, m), 3.98 (3H, m ), 5.65 (1H, d), 7.15-7.34 (11H, m), 7.40 (1H, d), 7.51 (2H, m).
[1212] Reference Example 79 Acid (S) -2- {4-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1benzylam¡no } -4-methylpentanoic
[1216] From Example 3. LCMS purity 100%, m / z 452 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 0.81 (6H, m), 1.51 (2H, m ), 1.72 (1H, m), 3.95 (2H, m), 5.61 (1H, d), 7.21-7.30 (4H, m), 7.39 (1H , d), 7.48 (2H, m), 7.52 (2H, m).
[1217] Reference Example 80 Acid (S) -2-l4-r6-am¡no-5- (3-methyl-4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1 -yl1-benzylamino} -3-phenylpropionic
[1221] From Example 5. LCMS purity 100%, m / z 500 [M + H] +, 1H NMR (400 MHz, DMSO), 5: 2.33 (3H, s), 2.90-3.07 (3H, m), 3.76 (1H, d), 3.92 (1H, d), 5.72 (1H, d), 7.20-7.42 (9H, m), 7 , 49 (4H, m).
[1222] Reference Example 81 Acid (S) -2-14-r6-am¡no-5- (3-methyl-4-fluorobenzo¡n-2-oxo-2H-P¡r¡d¡n-1- L1-benzylamino} -4-methylpentanoic
[1226] From Example 6. LCMS purity 98%, m / z 500 [M + H] +.
[1227] Reference Example 82 Cyclopentyl acid (S) -2- (3-14-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-yl1 -phenoxy} propylamine) -3-phenylpropionic
[1231] From Example 24. LCMS purity 93%, m / z 530 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 1.90 (2H, m), 2.80-2 , 90 (2H, m), 3.00 (2H, m), 3.40 (1H, m), 4.05 (2H, m), 5.70 (1H, d), 7.10 (1H , d), 7.20 (2H, d). 7.30 (5H, m), 7.35 (1H, d), 7.45 (1H, d), 7.60 (1H, d).
[1232] Reference Example 83 Acid (S) -2- (3- {4-r6-am¡no-5- (3-methyl-4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d ¡N-1-yl1phenox¡} -propylamino) -3-phenylpropionic
[1236] From Example 12. LCMS purity 96%, m / z 544 [M + H] ^{+, 1} H NMR (400 MHz, d ⁶ -DMSO), 5: 2.30 (2H, m), 2.40 (3H, s), 3.30 (1H, m), 4.30 (2H, m), 4.45 (1H, m), 5.85 (1H, d), 5.40 (1H , m), 5.85 (1H, d), 7.25 (2H, d), 7.40-7.55 (9H, m), 7.60-7.70 (2H, m).
[1237] Reference Example 84 Acid (SH3-14-r6-amino-5- (2,4-d¡fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-M1-phenox¡} -propylamino ) phenylacetic
[1241] From Example 28. LCMS purity 82%, m / z 534 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 5: 2.25 (2H, m), 3.10 (1H , m), 3.25 (1H, m), 4.20 (1H, m), 5.83 (1H, d), 7.15-7.60 (13H, d).
[1242] Reference Example 85 Acid (S) -2- (3-l4-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡l1fenox ¡} -Propylamino) -4-methylpentanoic
[1246] From Example 25. LCMS purity 100%, m / z 496 [M + H] +, 1 H NMR (400 MHz, d6-DMSO) 5: 1.00 ^(6H, m), 1,75- 1.90 (3H, m), 2.30 (2H, m), 3.10-3.30 (2H, m), 4.00 (1H, m), 4.25 (2H, m), 5 .85 (1H, d), 5.40 (1H, m), 5.85 (1H, d), 7.20 (2H, d), 7.30 (2H, d), 7.40 (2H, t), 7.55 (1H, m), 7.65 (2H, m).
[1247] Reference Example 86 Acid (S) -2- (3-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1- yl1phenoxy} -propylamine) -3-phenylpropionic
[1251] From Example 29. LCMS purity 100%, m / z 548 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 2.15 (2H, m), 3.15-3 , 30 (3H, m), 3.35 (1H, m), 4.10 (2H, m), 4.20 (1H, m), 5.65 (1H, d), 7.15 (2H, d), 7.20-7.35 (11H, m).
[1252] Reference Example 87 Acid (S) - (3-14-r6-am¡no-5- (4-fluoro-3-methylbenzo¡l) -2-oxo-2H-p¡r¡din-1- ¡L-phenoxy¡} -propylamine) phenylacetic
[1256] From Example 26. LCMS 95% purity, m / z 530 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 2.15 (3H, s) -2.35 (2H , m), 2.85 (2H, m), 3.05 (2H, m), 4.10 (2H, m), 5.25 (1H, m), 5.70 (1H, d), 4 , 25 (2H, m), 5.85 (1H, d), 5.40 (1H, m), 5.85 (1H, d), 7.10 (2H, d), 7.25 ( 2H, d), 7.3 (1H, d), 7.35 (1H, m), 7.40-7.55 (5H, m), 7.60 (2H, m).
[1257] Reference Example 88 Acid (S) -2- (3- {4-r6-am¡no-5- (4-fluoro-3-methylbenzo¡l) -2-oxo-2H-p¡r¡d ¡N-1-¡llphenox¡} -propylamino) -4-methylpentanoic
[1262] From Example 27. LCMS purity 94%, m / z 510 [M + H] +, 1H NMR (400 MHz, da-DMSO), 5: 0.90 (6H, bs), 1.65-1 , 80 (3H, m), 2.1-2.30 (3H, s + 2H, m), 3.0-3.20 (2H, m), 3.90 (1H, m), 4.15 ( 2H, m), 5.65 (1H, d), 7.15 (2H, d), 7.20-7.30 (3H, m), 7.30 (1H, m), 7.40 (1H , d), 7.45 (2H, s).
[1264] Reference Example 89 Acid S) - (3-l4-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-¡ll-3.5- difluorophenoxy} -propylamine) phenylacetic
[1269] From Example 13. LCMS purity 91%, m / z 552 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 2.10-2.25 (2H, ma), 2 , 80 (1H, m), 3.00 (1H, m), 4.15 (2H, d), 5.20 (1H, s), 5.70 (1H, d), 5, 65 (1H, d), 6.95 (2H, d), 7.30 (2H, t), 7.30 (1H, m), 7.40-7.60 (8H, m).
[1271] ^{Reference Example 90 Acid (SH3-14-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1- ¡n -3.5-} difluorophenoxy} propylam ino) phenylacetic
[1276] From Example 14. LCMS purity 98%, m / z 566 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 2.35 (2H, m), 3.1-3 , 3 (3H, m), 3.50 (1H, m), 4.25-4.40 (3H, m), 5.80 (1H, d), 5.70 (1H, d), 7, 10 (2H, d), 7.30-7.45 (7H, m), 7.60-7.70 (3H, m).
[1278] Reference Example 91 Acid (S) -2- (3- {4-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡din-1-¡ll- 3.5-difluorophenoxyphenoxy} propylamine) -4-methylpentanoic
[1283] From Example 15. LCMS purity 92%, m / z 532 [M + H] ^{+, 1} H NMR (400 MHz, d ⁶ -DMSO), 5: 0.95 (6H, m), 1.8 (3H, m), 2.30 (2H, m), 3.10-3.25 (2H, m), 3.95 (1H, m), 4.25 (2H, m), 5.80 ( 1H, d), 7.10 (2H, m), 7.40 (2H, m), 7.60 (1H, m), 7.65 (2H, m).
[1284] Reference Example 92 Acid (S) -2-l4-r6-am¡no-5- (2.4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-3.5-d ¡Fluorobenzylamino} -3-phenylpropionic
[1289] From Example 10. LCEM 95% purity, m / z 522 [M + H] +, 1H NMR (400 MHz, CD ³ OD), 5: 3.30 ( 2H, m), 4.15 (1H , m), 4.25 (2H, m), 5.75 (1H, d), 7.15-7.35 (9H, m), 5.20 (1H, m), 5.90 (1H , d), 7.35-7.50 (9H, m), 7.55 (2H, m), 7.65 (1H, d).
[1291] Reference Example 93 Acid (S) -2- (3-14-r6-am¡no-5- (4-fluoro-3-methylbenzo¡n-2-oxo-2H-p¡r¡d¡n -1-M1-3.5-d ifluorophenoxy} propylamine) -3-phenylpropionic
[1296] From Example 18. LCMS purity 92%, m / z 580 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 2.30 (2H, m), 2.40 (3H , s), 3.15-3.35 (4H, m), 3.50 (1H, m), 4.30 (2H, m), 4.35 (1H, m), 5.80 ( 1H, d), 7.10 (2H, m), 7.35-7.50 (7H, m), 7.55 (1H, m), 7.65 (1H, m).
[1298] Reference Example 94 (SH3-l4-r6-Amino-5- (4-fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-M1-phenoxy} propylamine) phenylacetic acid
[1303] From Example 23. LCMS purity 87%, m / z 516 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 2.25 (2H, m), 2.80 (1H , m), 3.10 (1H, m), 4.15 (2H, m), 5.30 (1H, s), 5.75 (2H, d), 7.15 (2H, d), 7 , 25 (2H, d), 7.40 (2H, t), 7.50-7.70 (7H, m).
[1304] Reference Example 95 Acid (SH3-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡n-2-oxo-2H-p¡r¡din-1-M1-3.5-d ifluorophenoxy} propylamine ) phenylacetic
[1309] From Example 20. LCMS purity 84%, m / z 570 [M + H] ^{+, 1} H NMR (400 MHz, d ⁶ -DMSO), 5: 2.30 (2H, m), 2.90 (1H, m), 3.15 (1H, m), 4.23 (2H, m), 5.32 (1H, s), 5.85 (1H, d), 7.10 (2H, d), 7.50 (3H, m), 7.60-7.65 (6H, m).
[1310] Reference Example 96 Acid (S) -2- (3-l4-r6-Am¡no-5- (2.4d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-illfenox ¡} -Propylamino) -4-methylpentanoic
[1315] From Example 30. LCMS purity 93%, m / z 514 [M + H] +, 1H NMR (400 MHz, da-DMSO), 5: 0.95 (6H, m), 1.85 (3H , m), 2.30 (2H, m), 3.15-3.22 (2H, m), 3.99 (1H, m), 4.21 (2H, m), 5.77 (1H , d), 7.20 (2H, d), 7.30 (4H, m), 7.45 (1H, m), 7.55 (1H, m).
[1317] Reference Example 97 Acid (S) -2- (3-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxo-2H-p¡r¡d¡n-1- ill-3.5-difluorophenoxy} -propylamino) -4-methylpentanoic
[1322] From Example 22. LCMS purity 88%, m / z 550 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 0.80-0.95 (6H, m), 1 , 50-1.85 (3H, m), 1.95-2.10 (2H, m), 2.95-3.05 (2H, m), 3.75-3.85 (1H, m) , 4.05-4.15 (2H, m), 5.65 (1H, d), 7.00 (1H, d), 7.10-7.20 (1H, m), 7.25-7 , 30 (1H, m), 7.30-7.40 (1H, m), 7.45-7.55 (1H, m), 7.80-8.25 (1H, bs), 9 , 90-10.20 (1H, bs).
[1324] Reference Example 98 Acid (S) -2- (3-14-r6-amino-5- (4-fluoro-3-methylbenzol) -2-oxo-2H-pyridin-1-ill- 3.5-difluorophenoxy} propylamino) -4-methylpentanoic
[1329] From Example 19. LCMS purity 93%, m / z 546 [M + H] +, 1H NMR (400 MHz, d6-DMSO), 5: 0.95-1.05 (6H, m), 1 , 55-1.75 (2H, m), 1.80-1.90 (1H, m), 2.10-2.25 (2H, m), 2.35 (3H, s), 3.00 -3.15 (2H, m), 3.70 (1H, m), 4.15-4.30 (2H, m), 5.80 (1H, d), 7.10 (1H, d), 7.25-7.35 (1H, m), 7.40-7.45 (1H, m), 7.50-7.55 (1H, m), 7.55-7.65 (1H, m ), 8.90-10.70 (2H, bs).
[1331] Reference Example 99 Acid (SH3-14-r6-Am¡no-5- (4-fluoro-3-metMbenzo¡l) -2-oxo-2H-p¡r¡d¡n-1-ill-3.5- difluorophenoxy} propylamine) phenylacetic
[1336] From Example 17. LCMS purity 100%, m / z 566 [M + H] ^{+, 1} H NMR (400 MHz, d ⁶ -DMSO), 5: 2.05-2.20 (2H, m) , 2.30 (3H, s), 2.75-3.05 (2H, m), 4.05-4.20 (2H, m), 4.65-4.85 (1H, m), 5.70 (1H, d), 7.00 (1H, d), 7.25-7.35 (1H, m), 7.35 to 7.60 ⁽⁸ H, m).
[1337] Reference Example 100 Acid (S) -2- (3-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡n-2-oxo-2H-p¡r¡din-1-¡l1- 3.5-Difluorophenoxy} -propylamino) -3-phenylpropionic
[1341] From Example 21. LCMS purity 100%, m / z 584 [M + H] +, 1H NMR (400 MHz, DMSO), ⁶ : 2.05-2.15 (2H, m) 3.00- 3.10 (3H, m), 4.00 to 4.25 (4H, m), 5.75 (1H, d), 7.05 (1H, d), 7.25-7.50 ⁽⁸ H , m), 7.55-7.65 (1H, m).
[1342] Reference Example 101 Acid (S) -2-l4-r6-m¡no-5- (4-fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-M1-3.5-d¡ fluorobenzylamino} -4-methylpentanoic
[1346] From Example 11. LCMS purity 92%, m / z 506 [M + H] +, 1 H NMR (400 MHz, MeOD) ^6: 0.85 to 1.05 ^(6H, m), 1, 65-1.85 (3H, m), 3.95-4.05 (1H, m), 4.25-4.35 (2H, m), 5.75 (1H, d), 6, 90 (1H, d), 7.00-7.10 (2H, m), 7.35-7.45 (4H, m).
[1347] Reference Example 102 Acid (S) -2- (4-14-r6-am¡no-5- (4-fluorobenzo¡n-2-oxo-2H-p¡r¡d¡n-1-M1-3.5 -difluorophenoxy} -cyclohexylamine) -4-methylpentanoic
[1351] From Example 31. LCMS purity 91%, m / z 606 [M ++ H] .1H NMR (400 MHz, CD ³ OD), ^6: 1.55 to 2.55 ⁽⁸ H, m), 3.20-3.40 (2H, m), 4.25-4.35 (1H, m), 4.45-4.55 (1H, m), 4.85-4.95 (1H , m), 5.95 (1H, d), 6.95-7.10 (2H, m), 7.35-7.55 ^(6H, m), 7.70-7.80 (2H, m), 7.80-7.85 (1H, d).
[1352] Exemp
[1356] From Example 32. LCMS purity 89%, m / z 592 [M + H] +, 1H NMR (400 MHz, CD ³ OD), ⁶ : 1.60-1.75 (2H, m), 1 , 80-1.95 (2H, m), 2.00-2.15 (2H, m), 2.15-2.30 (2H, m), 3.05-3.20 (1H, m) , 4.65-4.75 (1H, m), 4.75-4.80 (1H, m), 5.80 (1H, d), 6.85-6.95 (2H, m), 7 , 20-7.30 (2H, m), 7.40-7.50 (3H, m), 7.55-7.65 (4H, m), 7.65-7.70 (1H, m ).
[1357] Reference Example 104 N- (4-l4-r6-amino-5- (4-fluorobenzol) -2-oxop¡r¡din-1 (2H) -M1-3.5-difluorophenoxy} cyclohexih-L- leucine
[1362] From Example 33. LCMS purity 93%, m / z 572 [M + H] +, 1 H NMR (400 MHz, CD ³ OD), ^8: 0.85 to 1.00 ^(6H, m), 1.45-2.00 (9H, m), 2.05-2.25 (3H, m), 3.05-3.15 (1H, m), 3.60-3.75 (1H, m ), 4.30 and 4.65 (each 0.5H, m), 5.70 (1H, d), 6.75-6.85 (2H, m), 7.10-7.15 (2H , m), 7.45-7.55 (2H, m), 7.55-7.65 (1H, m).
[1364] Reference Example 105 (S ^ -M -re -am ino ^ - ^ - fluo robenzo in ^ -oxo ^ H -p irid in -l-illc ic lohexM -amino} -3-phenylpropionic acid
[1369] From Example 34. LCMS purity 98%, m / z 478 [M + H] +, 1H NMR (400 MHz, CD ³ OD), ⁸ : 1.55-1.85 (4H, m), 1 , 95-2.20 (2H, m), 2.30-2.75 (2H, m), 3.15-3.20 (1H, m), 3.25-3.35 (2H, m ), 4.05-4.15 (1H, m), 5.60 (1H, d), 7.05-7.15 (2H, m), 7.15-7.35 (5H, m ), 7.35-7.45 (3H, m).
[1371] Reference Example 106 Phenylacetic acid (R t-fó -M -re -am ino ^ -te ^ -d ifluo robenzo in ^ -oxo ^ H -p irid in -l-M I ^^ - d ifluorophenoxy} -propylamine)
[1376] From Example 42. LCMS purity ⁸⁸ %, m / z 570 [M + H] +, 1H NMR (400 MHz, DMSO), ⁸ : 2.05-2.20 (2H, m), 2.75 -2.95 (2H, m), 4.10-4.20 (2H, m), 4.30-4.50 (1H, m), 5.75 (1H, d), 7.00-7 , 10 (2H, m), 7.20-7.30 (1H, m), 7.35-7.50 (7H, m), 7.55-7.65 (1H, m).
[1378] Reference Example 107 Acid (2S) - ^ (3-14- ^ 6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -¡ ^ -3.5-Difluorophenoxy} propinamino1 (cyclohexyl) acetic
[1379] From Example 48, LCMS Purity 95%, m / z 576 [M + H] +, 1H NMR (300 MHz, DMSO), 8: 10.16 (1H, bs), 8.78 (1H , bs), 8.12 (1H, bs), 7.62-7.53 (1H, m), 7.47-7.32 (2H, m), 7.27-7.21 (1H , m), 7.07 (2H, d, J = 10.2 Hz), 5.74 (1H, d, J = 9.6 Hz), 4.19 (2H, t, J = 5.7 Hz ), 3.85-3.75 (1H, m), 3.15-3.00 (2H, m), 2.20-2.05 (2H, m), 1.95-1.60 (6H , m), 1.40-0.90 (5H, m).
[1381] Reference Example 108 N- (3-l4-r6-amino-5- (2,4-d¡fluorobenzo¡l) -2-oxop¡r¡din-1 (2H) -¡l1-3,5-difluorophenoxy } propyl) -D-leucine
[1386] From Example 50. CLEM purity 90%, m / z 550 [M + H] +, 1H NMR (300 MHz, DMSO), 8: 7.63-7.52 (1H, m), 7.46 -7.32 (2H, m), 7.28-7.10 (1H, m), 7.06 (2H, d, J = 10.2 Hz), 5.73 (1H, d, J = 9.9 Hz), 4.25-4.15 (2H, m), 3.90-3.80 (1H, m), 3.20-3.00 (2H, m), 2.20- 2.05 (2H, m), 1.80-1.55 (3H, m), 0.98-0.90 (6H, m).
[1388] Reference Example 109 N- (5-l4-r6-am¡no-5- (4-fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -M1-3.5-d¡fluorophenox¡ } -pentyl) -L-leuc¡na
[1393] From Example 55. To a solution of W- (5- {4- [6-amino-5- (4-fluorobenzoyl) -2-oxopyridin-1 (2H) -yl] -3,5-difluorophenoxy} pentyl ) -L-cyclopentyl leuccinate (33 mg, 0.05 mmol) in THF (1 ml) and water (1 ml) was added to Lio H (25 mg, 1.05 mmol, 20 equiv.). The mixture was stirred at room temperature for 16 hours, before being heated to 80 ° C for 10 hours. The mixture was concentrated under reduced pressure and water (5 ml) was added. The pH was adjusted to 7 using 1M HCl and the aqueous layer was extracted with 1-butanol (3 x 5 ml). The combined organic extracts were concentrated under reduced pressure. The solid residue was triturated with Et ² O, collected by filtration and purified by preparative HPLC to provide the title compound as a white solid as the mono-TFA salt (7 mg, 24 %). LC / MS: m / z 560 [M + H] +. 1H NMR (300MHz, DMSO-d6) 8: 7.62-7.51 (3H, m), 7.34 (2H, m), 7.05 (2H, m), 5.72 (1H, d, J = 9.8 Hz), 4.09 (2H, t, J = 5.7 Hz), 3.23 (1H, m), 2.80 (2H, m), 1.79-1 , 43 (9H, m), 0.89 (6H, t, J = 6.7Hz).
[1395] Reference Example 110 N- (l4-r6-Amino-5- (4-fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -M1phenox¡} -acetyl) -L -leucine
[1400] From Example________ 56. To a solution of W - ({4- [6-amino-5- (4-fluorobenzoyl) -2-oxopyridin-1 (2H) -yl] phenoxy} acetyl) -L-cyclopentyl leucinate (35 mg, 0.06 mmol) in THF (1 ml) and water (1 ml) was added to L¡Oh (30 mg, 1.24 mmol, 20 equiv.). The mixture was stirred at room temperature for 16 hours, concentrated under reduced pressure, and water (5 ml) was added. The pH was adjusted to 7 using 1M HCl and the aqueous layer was extracted with 1-butanol (3 x 5 ml). The combined organic extracts were concentrated under reduced pressure. The solid residue was triturated with Et ² O, filtered and dried under reduced pressure to provide the title compound as a cream colored solid (11 mg, 36% yield).
[1401] LC / MS: m / z 496 [M + H] +. 1H NMR (300 MHz, DMSO-d6) 8: 7.58-7.53 (3H, m), 7.43 (1H, d, J = 9.6 Hz), 7.36-6.98 ( 6H, m), 5.68 (1H, d, J = 9.6Hz), 4.58 (2H, s), 3.90 (1H, m), 1.67-1.31 (3H , m), 0.86 (6H, m).
[1402] ^{Reference Example 111 N-r2- (4-16-am¡no-5-r (4-fluorophenyl) carbon¡} n ^{-2-oxop¡r¡d¡n-1 (2H) -¡l} fen ¡L) etM1-L-leucine}
[1406] From Example 58. LC / MS: m / z 466 [M + H] +. 1H NMR (300 MHz, DMSO-d ⁶ ) 5: 8.21 (1H, bs), 7.60-7.44 (4H, m), 7.39-7.30 (4H, m), 5, 76-5.69 (1H, m), 4.00-3.85 (1H, m), 3.10-2.95 (2H, m), 1.85-1.60 (3H, m ), 1.30-1.10 (2H, m), 0.95 ^(6H, d, J = ^6Hz).
[1407] Reference Example 112 N-r2- (4-16-amino-5-r (4-methylphenyl) carbonM1-2-oxop¡r¡d¡n-1 (2H) -¡l} fen ¡L) et¡l1-L-leuc¡na
[1411] From Example 63. LC / MS: m / z 462 [M + H] +. 1H NMR (300 MHz, CD ³ OD) 5: 7.69 (1H, d, J = 9.6 Hz), 7.53 (2H, d, J = 7.2 Hz), 7.45 (2H, d, J = 8.1 Hz), 7.34 (2H, d, J = 7.8 Hz), 7.25 (2H, d, J = 8.4 Hz), 5.80 (1H, d, J = 9.6 Hz), 3.15 (1H, m), 3.02-2.75 (4H, m), 2.45 (3H, s), 1.73 (1H, m), 1 , 56 to 1.22 (2H, m), 0.96 ^(6H, dd).
[1412] Reference Example 113 N-r2- (4-16-am¡no-5-r (4-methoxyphen¡l) carbonM1-2-oxop¡r¡d¡n-1 (2H) -¡l} - fen¡l) et¡l1-L-leuc¡na
[1416] From Example 64. LC / MS: m / z 478 [M + H] +. 1H NMR (300 MHz, DMSO-d6) 5: 7.28 (2H, d, J = 8.7 Hz), 7.15 (2H, d, J = 8.1 Hz), 7.04 (1H, d, J = 9.3 Hz), 6.89 (4H, m), 4.87 (1H, d, J = 9.3 Hz), 3.78 (1H, m), 3.41 (3H, s), 2.75 (2H, m), 1.78 (1H, m), 1.24 (2H, m), 0.86 ^(6H, t).
[1417] Reference Example 114 N-r2- (4-16-am¡no-5-r (4-chlorophen¡l) carbon¡l1-2-oxop¡r¡d¡n-1 (2H) -¡l} fen ¡L) et¡l1-L-leucine
[1421] From Example 65. LC / MS: m / z 482 [M + H] +. 1H NMR (300 MHz, CD ³ OD) 5: 7.16 (1H, d), 7.52 ^(6H, m), 7.23 (2H, d), 6.82 (1H, d), 3.15 (1H, t), 1.74 (1H, m), 1.44 (2H, m), 0.93 ^(6H, dd).
[1422] Reference Example 115 (2S) -4-am¡no-2-r (3-l4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡din-1 ( 2H) -¡l1-3.5-d¡fluorophenox¡} prop¡l) amino1butanoate
[1427] From Example 52. LC / MS: m / z 537 [M + H] +. 1H NMR (300MHz, DMSO-da) 5: 7.52-7.75 ( 2H, m), 7.30-7.48 (2H, m), 7.20-7.29 (1H, m) , 7.09 (2H, d, J = 9.7Hz), 4.23 (1H, t, J = 6.1Hz), 4.07-4.17 (2H, m), 2.14 - 2.32 (2H, m), 1.22-1.41 (6H, m), 0.88 (4H, t, J = 7.3 Hz)
[1429] Reference Example 116 N- (5-f4-r6-am¡no-5- (2.4-d¡fluorobenzo¡l) -2-oxop¡r¡d¡n-1 (2H) -il1-3.5-d¡ fluorophenoxy} -pentyl) -L-leucine
[1434] From Example 54. To a solution of W- (5- {4- [6-amino-5- (2,4-difluorobenzoyl) -2-oxopyridin-1 (2H) -yl] -3,5-difluorophenoxy } pentyl) -L-tert-butyl leucinate (21 mg, 0.04 mmol) in DCM (2.5 ml) was added to TFA (2.5 ml). The mixture was stirred at room temperature for 20 hours, before being concentrated under reduced pressure. The residue was dissolved in minimal MeOH and azeotroped three times with 1: 1 toluene / DCM. The title compound was provided as a cream colored solid as the mono-TFA salt (21 mg, 92% yield). LC / MS: m / z 634 [M + H] +. 1H NMR (300MHz, DMSO-d6) 5: 10.14 (1H, bs), 8.21 (1H, bs), 7.57 (1H, m), 7.46 (1H, m ), 7.34 (1H, dd, J = 9.6, 2.4 Hz), 7.21 (1H, m), 7.06 (2H, d, J = 10.2 Hz), 5 , 73 (1H, d, J = 9.9Hz), 4.10 (2H, t, J = 5.7Hz), 3.40 (1H, m), 2.84 (2H, t, J = 6.6 Hz), 1.79-1.48 (9H, m), 0.90 (6H, t, J = 6.3 Hz).
[1436] The following examples were prepared in a similar manner:
[1438] Reference Example 117 N-r2- (4-f6-am¡no-5-r (2.4-d¡fluorophen¡l) carbonM1-2-oxop¡r¡d¡n-1 (2H) -¡l} fen ¡L) -et¡l1-L-leuc¡na
[1443] From Example 60. LC / MS: m / z 484 [M + H] +: 1H NMR (300 MHz, DMSO-d6) 5: 10.06 (1H, bs), 9.17 (2H, bs ), 7.55-6.94 (8H, m), 5.72 (1H, d, J = 9.6 Hz), 4.05-3.93 (1H, m), 3.40-3, 10 (3H, m), 1.85-1.65 (4H, m), 0.95 (6H, d, J = 5.7 Hz).
[1445] Reference Example 118 Acid (2SHr2- (4-16-amino-5-r (4-fluorophenyl) carbonM1-2-oxop¡r¡d¡n-1 (2H) -¡l) phen¡l ) etM1-amino} (phenyl) ethanoic
[1448] From Example 62. LC / MS: m / z 486 [M + H] +, 1H NMR (300 MHz, DMSO-d6) 8: 9.80 (2H, bs), 7.70-7.20 ( 14H, m), 5.70 (1H, d, J = 9.6Hz), 5.24 (1H, s), 3.20-2.90 (4H, m).
[1450] Measurement of biological activities
[1452] P38 MAP kinase activity
[1454] The ability of compounds to inhibit MAP kinase activity at p38 was measured in an assay carried out by Upstate (Dundee, UK). In a final reaction volume of 25 µl, p38 MAP kinase (5-10 mU) was incubated with 25 mM Tris, pH 7.5, 0.002 mM EGTA, 0.33 mg / ml myelin basic protein, acetate 10 mM Mg and [g-33p-ATP] (specific activity approximately 500 cpm / pmol, concentration as required). The reaction was started by adding the MgATP mixture. After incubating for 40 minutes at room temperature, the reaction was stopped by adding 5 µl of a 3% phosphoric acid solution. Then 1 µl of the reaction was added onto a Filtermat P30 and washed three times for 5 minutes in 75 mM phosphoric acid and once in methanol before drying and scintillation counting.
[1456] Duplicate data points were generated from a log 1/3 serial dilution of a stock solution in DMSO. Nine dilution steps are carried out starting from a maximum concentration of 10 pM, and a "no compound" blank is included. The radiometric filter binding assay was carried out at an ATP concentration of or near km. Scintillation count data was collected and subjected to free fit analysis using Prism software. From the curve generated, the concentration that provided 50% inhibition was determined and reported.
[1458] LPS stimulation of THP-1 cells
[1460] THP-1 cells were seeded in 100 µl at a density of 4 x 104 cells / well in 96-well V-bottom tissue culture treated plates and incubated at 37 ° C in 5% CO ² for 16 h. 2 h after the addition of the inhibitor in 100 µl of tissue culture medium, the cells were stimulated with LPS ( E coli strain 005: B5, Sigma) at a final concentration of 1 pg / ml and incubated at 37 ° C in 5% CO ² for 6 h. TNF-α levels from cell-free supernatants were measured by sandwich ELISA (R&D Systems # QTA00B).
[1462] LPS stimulation of human whole blood
[1464] Whole blood was drawn by venipuncture using heparinized vacutainers (Becton Dickinson) and diluted in an equal volume of RPMI1640 tissue culture medium (Sigma). 100 µl was plated into 96-well V-bottom tissue culture treated plates. 2 h after the addition of the inhibitor in 100 µl of RPMI1640 medium, the blood was stimulated with LPS ( E. coli strain 005: B5, Sigma) at a final concentration of 100 ng / ml and incubated at 37 ° C in 5% CO ² for 6 h. TNF-α levels from cell-free supernatants were measured by sandwich ELISA (R&D Systems # QTA00B).
[1466] IC50 values were assigned to one of three ranges as follows:
[1468] Range A: IC50 <100 nM
[1469] Range B: 100 nM <IC50 <1000 nM
[1470] Range C: IC50> 1000 nM
[1472] Results table
[1474]
[1475]
[1476]
[1477]
[1480] Lysed Cell Carboxylesterase Assay
[1482] Any given compound of the present invention in which Ri is an ester group can be tested to determine whether it meets the requirement of being hydrolyzed by intracellular esterases, by testing with the following assay.
[1484] Cell extract preparation
[1486] U937 or Hut78 tumor cells (~ 109) were washed in 4 volumes of Dulbecco's PBS (~ 1 liter) and pelleted at 525 g for 10 min at 4 ° C. This was repeated twice and the final cell pellet was resuspended in 35 ml of cold homogenization buffer (10 mM Trizma, 130 mM NaCl, 0.5 mM CaCb, pH 7.0 at 25 ° C). Homogenates were prepared by cavitation in nitrogen (4826 kPa (700 psi) for 50 min at 4 ° C). The homogenate was kept on ice and supplemented with a cocktail of inhibitors at final concentrations of:
[1488] Leupeptin 1 pM
[1489] Aprotinin 0.1 pM
[1490] E64 8 pM
[1491] Pepstatin 1.5 pM
[1492] Bestatin 162 pM
[1493] Chymostatin 33 pM
[1495] After clarification of the cell homogenate by centrifugation at 525 g for 10 min, the resulting supernatant was used as a source of esterase activity and stored at -80 ° C until needed.
[1497] Ester cleavage measurement
[1499] The hydrolysis of the esters to the corresponding carboxylic acids can be measured using the cell extract, prepared as in the previous case. For this effect, the cell extract (~ 30 pg / 0.5 ml total test volume) was incubated at 37 ° C in a buffer of 25 mM Tris-HCl, 125 mM NaCl, pH 7.5 at 25 ° C . Then, at time zero, the ester (substrate) was added to a final concentration of 2.5 pM and the samples were incubated at 37 ° C for a suitable time (usually 0 or 80 min). Reactions were stopped by adding 3 volumes of acetonitrile. For samples at time zero, the acetonitrile was added before the ester compound. After centrifuging at 12000 g for 5 min, the samples were analyzed for the ester and its corresponding carboxylic acid at room temperature by LCMS (Sciex API 3000, HP1100 binary pump, CTC PAL). Chromatography was based on an AceCN column (75 x 2.1 mm) and a mobile phase of acetonitrile in 5-95% water / 0.1% formic acid.
[1501] Hydrolysis rates are expressed in pg / ml / min.
[1503] Table 1 presents data showing that various amino acid ester motifs, conjugated to various intracellular enzyme inhibitors by several different binding chemistries are all hydrolyzed by intracellular carboxyesterases to the corresponding acid.
[1504]

权利要求:
Claims (13)
[1]
1. A compound of formula (I):

[2]
2. A compound according to claim i, wherein P is hydrogen and U is a radical of formula (IA) as defined in claim i.
[3]
A compound according to any preceding claim, wherein A is optionally substituted i, 4-phenylene.
[4]
4. A compound according to claim i having the formula (IIA), (IIB) and (IIC):

[5]
5. A compound according to any of claims 1 to 4, wherein the radical -Y-L1-X1- [CH ² ] z-, is -CH ² -, -CH ² CH ² -, -CH ² CH ² CH ² -, -CH ² CH ² CH ² CH ² -, -CH ² O-, -CH ² CH ² O-, -CH ² CH ² CH ² O-, -CH ² CH ² CH ² CH ² O-, -C (= O) -CH ² -, - C (= O) -CH ² O-, -C (= O) -NH-CH ² - or -C (= O) -NH-CH ² OR-.
[6]
6. A compound according to any preceding claim, wherein R ¹ is an ester group of formula - (C = O) OR14, wherein R ¹⁴ is R ⁸ R ⁹ R ¹⁰ C- where
(i) R8 is hydrogen or (C1-C3) alkyl - (Z1) a- [(C1-C3) alkyl] b- or (C2-C3) alkenyl - (Z1) a- [(C1-C3) alkyl] b- optionally substituted, where a and b are independently 0 or 1 and Z1 is -O-, -S- or -NR ¹¹ - where R ¹¹ is hydrogen or (C ¹ -C ³ ) alkyl; and R ⁹ and R ¹⁰ are independently hydrogen or (C ¹ -C ³ ) alkyl -;
(ii) R8 is hydrogen or R12R13N-(C ¹ -C ³ ) alkyl - optionally substituted where R ¹² is hydrogen or (C ¹ -C ³ ) alkyl and R ¹³ is hydrogen or (C ¹ -C ³ ) alkyl; or R ¹² and R ¹³ , together with the nitrogen to which they are attached, form an optionally substituted monocyclic heterocyclic ring of 5 or 6 ring atoms or heterocyclic bicyclic ring system of 8 to 10 ring atoms, and R ⁹ and R ¹⁰ are independently hydrogen or (C ¹ -C ³ ) alkyl -; or (iii) R8 and R ⁹ , taken together with the carbon to which they are attached, form an optionally substituted monocyclic carbocyclic ring of 3 to 7 ring atoms or bicyclic carbocyclic ring system of 8 to 10 ring atoms, and R ¹⁰ is hydrogen.
[7]
7. A compound according to claim 6, wherein R ¹⁴ is methyl, ethyl, n- or iso-propyl, n-, sec- or tert-butyl, cyclohexyl, allyl, phenyl, benzyl, 2-, 3 - or 4-pyridylmethyl, N-methylpiperidin-4-yl, tetrahydrofuran-3-yl or methoxyethyl.
[8]
8. A compound according to any preceding claim, wherein R ² is phenyl, benzyl, iso-butyl, cyclohexyl or t-butoxymethyl.
[9]
9. A compound according to any of claims 1 to 5, wherein R ¹ is an ester group of formula - (C = O) OR14, wherein R ¹⁴ is cyclopentyl, and R ² is phenyl, benzyl, iso-butyl, cyclohexyl, or t-butoxymethyl.
[10]
10. A compound according to claim 1 selected from the group consisting of
Cyclopentyl (S) - (3- {4- [6-Amino-5- (2,4-difluorobenzoyl) -2-oxo-2H-pyridin-1-yl] -3,5-difluorophenoxy} propylamino) phenylacetate
(S) -2- (3- {4- [6-Amino-5- (2,4-difluorobenzoyl) -2-oxo-2H-pyridin-1-yl] -3,5-difluorophenoxy} propylamino) -4 -cyclopentyl methylpentanoate
Example 42 (2R) - [(3- {4- [6-Amino-5- (2,4-difluorobenzoyl) -2-oxopyridin-1 (2H) -yl] -3,5-difluorophenoxy} propyl) amino] - (phenyl) cyclopentyl acetate
W- (3- {4- [6-amino-5- (2,4-difluorobenzoyl) -2-oxopyridine-1- (2H) -yl] -3,5-difluorophenoxy} propyl) -L-leucinate
2-morpholin-4-ylethyl
W- (3- {4- [6-amino-5- (2,4-difluorobenzoyl) -2-oxopyridine-1- (2H) -yl] -3,5-difluorophenoxy} propyl) -L-leucinate
2- (dimethylamino) ethyl
W- [2- (4- {6-am¡no-5 - [(4-fluorophenyl) carbon¡l] -2-oxop¡r¡d¡n-1 (2H) -¡l} phen¡ l) cyclopentyl ethyl] -L-leucinate
W- (5- {4- [6-am¡no-5- (2,4-d¡fluorobenzo¡l) -2-oxop¡nd¡n-1 (2H) -¡l] -3.5- d¡fluorophenox¡} pent¡l) -L-cyclopentyl leucinate W- [3- (4- {6-amino-5 - [(4-fluorophenyl) carbon¡l] -2-oxop¡r¡d¡n-1 (2H) -¡l} phen¡l) prop¡l] -L-cyclopent¡lo leucinate
(2S) -4-Amino-2 - [(3- {4- [6-amino-5- (2,4-d¡fluorobenzo¡l) -2-oxop¡nd¡n-1 ( Cyclopentyl 2H-¡l] -3,5-d¡fluorophenox¡} prop¡l) amino] -butanoate
W- (5- {4- [6-am¡no-5- (4-fluorobenzo¡l) -2-oxop¡nd¡n-1 (2H) -¡l] -3,5-d¡fluorophenox¡ } pentl) -L-cyclopentyl leucinate W- [2- (4- {6-amine-5 - [(2,4-dfluorophenyl) carbonl] -2-oxop¡r¡d¡n-1 (2H) -¡l} phen¡l) eth¡l] -L-cyclopent¡lo leucinate
W- [2- (4- {6-am¡no-5 - [(2,4-d¡fluorophen¡l) carbon¡l] -2-oxop¡r¡d¡n-1 (2H) -¡ l} phenyl) ethyl] -L-tert-butyl leucinate
(2S) - {[2- (4- {6-am¡no-5 - [(4-fluorophenyl) carbon¡l] -2-oxop¡r¡d¡n-1 (2H) -¡l } phenyl) ethyl] amino} (phenyl) cyclopentyl ethanoate W- [2- (4- {6-amino-5 - [(4-methylphene) l) carbon¡l] -2-oxop¡r¡d¡n-1 (2H) -¡l} phen¡l) eth¡l] -L-cyclopentyl leucinate
W- [2- (4- {6-am¡no-5 - [(4-chlorophenyl) carbon¡l] -2-oxop¡r¡d¡n-1 (2H) -¡l} phen¡ l) ethyl] -L-cyclopentyl leucinate.
[11]
11. A compound in accordance with any one of the preceding revisions which is in the form of a pharmaceutically acceptable salt.
[12]
12. A pharmaceutical composition comprising a compound according to any one of the foregoing revisions, together with a pharmaceutically acceptable carrier.
[13]
13. A compound according to any one of revisions 1 to 10 for the treatment of an autoimmune or inflammatory disease.

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同族专利:

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公开号 | 公开日

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US8044211B2|2011-10-25|

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WO2007129040A1|2007-11-15|

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EP2013175A1|2009-01-14|

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NZ573348A|2012-01-12|

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ES2605727T3|2017-03-16|

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CN102942520B|2015-03-04|

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BRPI0711310B1|2020-12-15|

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JP2009535385A|2009-10-01|

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CA2650970C|2014-09-16|

---

CN102942520A|2013-02-27|

---

DK2013175T3|2017-02-20|

---

EP2013175B1|2016-11-09|

---

CY1118518T1|2017-07-12|

---

BRPI0711310B8|2021-05-25|

---

HUE033281T2|2017-11-28|

---

PT2013175T|2017-02-14|

---

JP5340918B2|2013-11-13|

---

AU2007246869B2|2012-10-18|

---

AU2007246869A1|2007-11-15|

---

CA2650970A1|2007-11-15|

---

US20090099185A1|2009-04-16|

---

BRPI0711310A2|2011-12-06|

---

KR101429780B1|2014-08-18|

---

KR20090014187A|2009-02-06|

---

EP2013175B9|2020-11-11|

---

DK2013175T5|2020-12-21|

引用文献:

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IL163957D0|2002-03-14|2005-12-18|Bayer Healthcare Ag|Monocyclic aroylpyridinones as antiinflammatory agents|

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EP1964577B1|2005-05-05|2016-04-13|GlaxoSmithKline Intellectual Property Development Limited|Alpha aminoacid ester-drug conjugates hydrolysable by carboxylesterase|

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EP2245012B1|2008-02-29|2013-07-03|Chroma Therapeutics Limited|Inhibitors of p38 map kinase|GB0619753D0|2006-10-06|2006-11-15|Chroma Therapeutics Ltd|Enzyme inhibitors|

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CA2668070A1|2006-10-30|2008-05-08|Chroma Therapeutics Ltd.|Hydroxamates as inhibitors of histone deacetylase|

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CN101842361B|2007-06-27|2013-06-05|阿斯利康有限公司|Pyrazinone derivatives and their use in the treatment of lung diseases|

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EP2220044A1|2007-11-07|2010-08-25|Chroma Therapeutics Limited|P38 map kinase inhibitors|

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GB0803747D0|2008-02-29|2008-04-09|Martin|Enzyme and receptor modulation|

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EP2245012B1|2008-02-29|2013-07-03|Chroma Therapeutics Limited|Inhibitors of p38 map kinase|

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CN102099036B|2008-06-03|2015-05-27|英特芒尼公司|Compounds and methods for treating inflammatory and fibrotic disorders|

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AR073711A1|2008-10-01|2010-11-24|Astrazeneca Ab|ISOQUINOLINE DERIVATIVES|

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GB0903480D0|2009-02-27|2009-04-08|Chroma Therapeutics Ltd|Enzyme Inhibitors|

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GB201211310D0|2012-06-26|2012-08-08|Chroma Therapeutics Ltd|CSF-1R kinase inhibitors|

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AR092742A1|2012-10-02|2015-04-29|Intermune Inc|ANTIFIBROTIC PYRIDINONES|

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RS59458B1|2012-10-17|2019-11-29|Macrophage Pharma Ltd|N-[2-{4-[6-amino-5--2-oxopyridin-1-yl]-3,5-difluorophenyl}ethyl]-l-alanine and the terbutyl ester thereof|

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GB201306901D0|2013-04-16|2013-05-29|Chroma Therapeutics Ltd|Combination|

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RU2692485C2|2014-04-02|2019-06-25|Интермьюн, Инк.|Antifibrous pyridinones|

法律状态:

优先权:

---

申请号 | 申请日 | 专利标题

---

GB0608855A|GB0608855D0|2006-05-04|2006-05-04|Inhibitors of MAP kinase enzymes|

---

GB0608855|2006-05-04|

---

GB0613914A|GB0613914D0|2006-07-13|2006-07-13|Inhibitors of p38 map kinase|

---

GB0613914|2006-07-13|

---

PCT/GB2007/001596|WO2007129040A1|2006-05-04|2007-05-01|p38 MAP KINASE INHIBITORS|

---