farnesoid x receptors, their uses, combination and pharmaceutical composition

专利摘要:
PHARNESOID X RECEPTORS, THEIR USES, PHARMACEUTICAL COMPOSITION, AND COMBINATION. The present invention relates to compounds of Formula I, (I) a stereoisomer, enantiomer, a pharmaceutically acceptable salt or an amino acid conjugate thereof; where variables are defined here; and their pharmaceutical compositions, which are useful as modulators of Farnesoid X (FXR) receptor activity.
公开号:BR112016009630B1
申请号:R112016009630-4
申请日:2014-11-04
公开日:2021-02-09
发明作者:Donatella Chianelli；Xiaodong Liu；Valentina Molteni；John Nelson；Jason ROLAND；Paul Rucker；David Tully
申请人:Novartis Ag；
IPC主号:

专利说明:

CROSS REFERENCE TO RELATED REQUESTS
[001] This application claims the benefit of provisional application serial number US 61 / 900,013, filed on November 5, 2013, which is incorporated by reference here in its entirety. FIELD OF THE INVENTION
[002] The present invention relates to compositions and methods for modulating the activity of farnesoid X receptors (FXRs). BACKGROUND OF THE INVENTION
[003] The farnesoid X receptor (FXR) is a member of the nuclear hormone receptor superfamily and is mainly expressed in the liver, kidney and intestine (see, for example, Seol et al. (1995) Mol. Endocrinol. 9: 72-85 and Forman et al. (1995) Cell 81: 687-693). Its functions as a heterodimer with the retinoid X receptor (RXR) and binds to response elements in target gene promoters to regulate transcription genes. The FXR-RXR heterodimer binds with high affinity to an inverted repeat 1 (IR-1) response element, in which the consensus receptor binding hexamers are separated by a nucleotide. FXR is part of an interrelated process, in which FXR is activated by bile acids (the end product of cholesterol metabolism) (see, for example, Makishima et al. (1999) Science 284: 1362-1365, Parks et al (1999) Science 284: 1365-1368, Wang et al. (1999) Mol. Cell. 3: 543-553), which serves to inhibit cholesterol metabolism. See also, Urizar et al. (2000) J. Biol. Chem. 275: 39313-39317.
[004] FXR is a key regulator of cholesterol homeostasis, triglyceride synthesis and lipogenesis. (Crawley, Expert Opinion Ther. Patents (2010), 20 (8): 1047-1057). In addition to the treatment of dyslipidemia, multiple indications for FXR have been described, including treatment of liver disease, diabetes, vitamin D-related diseases, drug-induced side effects and hepatitis. (Crawley, supra). Although advances have been made in the development of new FXR agonists, there is still significant room for improvement. SUMMARY OF THE INVENTION
[005] The present invention relates to compositions and methods for modulating the activity of farnesoid X receptors (FXRs). For example, the present invention provides new compounds that are FXR agonists or partial agonists, and are useful as pharmaceuticals for treating FXR-mediated conditions.
[006] In one aspect, the compounds of the invention are defined by Formula (I):

[007] or a pharmaceutically acceptable salt thereof, where,
[008] R0 is Ring A or C1-6 alkyl;
[009] Ring A is aryl; 5 to 10 membered heteroaryl comprising 1 to 3 N, O or S heteroatoms; 4- to 6-membered heterocycle comprising 1 to 2 N, O or S heteroatoms; or C3-7 cycloalkyl; and said Ring A is substituted or not by 1 to 2 substitutes independently represented by R2;
[0010] Ring B is aryl; 5 to 10 membered heteroaryl comprising 1 to 3 N, O or S heteroatoms; 4- to 6-membered heterocycle comprising 1 to 2 N, O or S heteroatoms; or C3-7 cycloalkyl; and said Ring B is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[0011] X is - (CR4R5) - or -C (O) -;
[0012] Y is -O-, - (CR4R5) -, * -O (CR4R5) - or -NR-, where "*" indicates the point of attachment of Y to the ring containing the ring atoms Z;
[0013] Z1, Z2, Z3, and Z4 are each independently -CR3- or -N-;
[0014] L1 is * 1– (CR4R5) 1-2– or * 1– (CR4R5) -C (O) -NR–, where "* 1" indicates the point of attachment from L1 to N;
[0015] L is - (CR R) 1-2-, - (CR R) -C (O) -, - (CR R) -C (O) - NR-, * 2- (CR4R5) 2-O -, * 2- (CR4R5) 2-NR-, * 2- (CR4R5) 2-SO2-, * 2- (CR4R5) 2-NR-C (O) -, or * 2- (CR4R5) -C ( O) -NR- (CR4R5) -; where "* 2" indicates the connection point from L2 to N;
[0016] L3 is - (CR4R5) - or -C (O) -;
[0017] each R2 is independently halo, hydroxyl, C1-6 alkyl, or C1-6 alkyl substituted by halo;
[0018] each R3 is independently hydrogen, halo, or C1-6 alkyl; and
[0019] R, R4 and R5 are independently hydrogen or C1-6 alkyl.
[0020] In one embodiment, the compounds of the invention are defined by Formula (II):
or a pharmaceutically acceptable salt thereof; on what,
[0021] R0 is Ring A or C1-6 alkyl; wherein Ring A is phenyl, pyridyl or cyclopropyl, each of which is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[0022] Ring B is selected from phenyl, pyridyl, 1H-indolyl-2 substitutes independently represented by R2;
[0023] X is - (CR4R5) -;
[0024] Y is -O-, - (CR4R5) - or * -O (CR4R5) -, where "*" indicates the point of attachment of Y to the ring containing the ring atoms Z;
[0025] Z1, Z2, Z3, and Z4 are each independently -CR3- or -N-;
[0026] L1 is * 1- (CR4R5) 1-2- where "* 1" indicates the point of attachment from L1 to N;
[0027] L2 is * 2- (CR4R5) 1-2-, * 2- (CR4R5) -C (O) -NR-, * 2- (CR4R5) 2-O-, * 2- (CR4R5) 2- NR- or * 2- (CR4R5) -C (O) -NR- (CR4R5) -; where "* 2" indicates the connection point from L2 to N;
[0028] each R2 is independently halo, C1-6 alkyl, or C1-6 alkyl substituted by halo;
[0029] each R3 is independently hydrogen, halo, or C1-6 alkyl; and
[0030] R, R4 and R5 are independently hydrogen or C1-6 alkyl.
[0031] In another embodiment, the compounds of the invention are represented by Formula (III):

[0032] or a pharmaceutically acceptable salt thereof; on what
[0033] Ring A is phenyl or pyridyl, each of which is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[0034] Ring B is selected from phenyl, pyridyl, 1H-indolyl, and cyclopentyl, each of which is irreplaceable or substituted by 1 to 2 substituents independently represented by R2;
[0035] L1 is - (CR4R5) -;
[0036] L2 is selected from - (CH2) -, * 2-CH2C (O) NH-, * 2- CH (CH3) C (O) NH-, * 2-CH2C (O) NHCH2-, * 2- (CH2) 2O-, and * 2- (CH2) 2NH-; where "* 2" indicates the connection point from L2 to N;
[0037] X is CH2;
[0038] Y is selected from -O-, -CH2-, -C (CH3) 2- and * -O-CH2-, where "*" indicates the point of attachment of Y to the ring containing the ring atoms Z ;
[0039] Z1 is CR3 or N;
[0040] Z2 is CR3;
[0041] Z3 is CR3;
[0042] Z4 is CR3 or N;
[0043] each R2 is independently selected from halo, methyl, and trifluoromethyl;
[0044] each R3 is independently hydrogen, halo, or C1-6 alkyl; and
[0045] each R4 and R5 is independently hydrogen or methyl.
[0046] In yet another embodiment, the compounds of the invention are represented by Formula (IV):

[0047] or a pharmaceutically acceptable salt thereof; on what
[0048] each phenyl ring is optionally substituted again by 1 to 2 substituents independently represented by R2, wherein R2 is fluorine or methyl;
[0049] L2 is selected from -CH2-, * 2-CH2CH2NH-, * 2-CH2CH2O-, and * 2-CH2C (O) NH-, where "* 2" indicates the point of attachment from L2 to N;
[0050] Z2 is selected from CH, CF, CCl, and CCH3; and
[0051] Z3 is selected from CH, CF, CCl, and CCH3.
[0052] The present invention also provides a compound represented by Formula (V):

[0053] in which each phenyl ring is optionally substituted again by 1-2 substituents independently represented by R2, where R2 is fluorine or methyl;
[0054] L2 is selected from -CH2-, * 2-CH2CH2NH-, * 2-CH2CH2O-, and * 2-CH2C (O) NH-, where "* 2" indicates the point of attachment from L2 to N;
[0055] Z2 is selected from CH, CF, CCl, and CCH3;
[0056] Z3 is selected from CH, CF, CCl, and CCH3; and
[0057] R6 is C1-6 alkyl.
[0058] The compounds of Formula I, II, III, IV and V, and their pharmaceutically acceptable salts exhibit valuable pharmacological properties when tested in vitro in cell-free kinase assays and in cell assays, and are therefore useful as products pharmacists.
[0059] In one aspect, the invention relates to methods for modulating FXR in a cell, comprising contacting the cell with an effective amount of a compound of Formula I, II, III, IV or V, or a pharmaceutically acceptable salt of the same; and optionally in combination with a second therapeutic agent.
[0060] In another aspect, the invention relates to methods for treating, ameliorating or preventing an FXR-mediated disorder in an individual suffering from, comprising administering to the individual a therapeutically effective amount of a compound of Formula I, II, III , IV or V, or a pharmaceutically acceptable salt thereof; and optionally in combination with a second therapeutic agent. The present invention also provides for the use of a compound of Formula I, II, III, IV or V, or a pharmaceutically acceptable salt thereof; and optionally in combination with a second therapeutic agent, in the manufacture of a medicament for treating an FXR-mediated disorder. In yet another aspect, the present invention relates to a combination comprising a therapeutically effective amount of a compound of Formula I, II, III, IV or V, or a pharmaceutically acceptable salt thereof, and a second therapeutic agent. therapeutic. Where a second therapeutic agent is used, the second therapeutic agent may also be useful in the treatment of an FXR-mediated disorder.
[0061] In one embodiment, the compounds (alone or in combination with a second therapeutic agent) are useful for treating liver disease or gastrointestinal disease, including, but not limited to, liver diseases selected from intrahepatic cholestasis, induced cholestasis estrogen, drug-induced cholestasis, cholestasis of pregnancy, cholestasis associated with parenteral nutrition, progressive family cholestasis (PFIC), Alagille syndrome, primary biliary cirrhosis (PBC), primary sclerosing cholangitis, ductopenic liver transplant rejection, graft disease versus host associated with liver transplantation, cystic fibrosis kidney disease, non-alcoholic grease liver disease (NAFLD), non-alcoholic steatoepatitis (NASH), alcoholic liver disease, and kidney disease associated with parenteral nutrition; and gastrointestinal diseases selected from bile acid malabsorption (including primary bile acid diarrhea and secondary bile acid diarrhea), bile reflux gastritis, and inflammatory bowel disease such as Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, fun colitis, indeterminate colitis and Behcet's disease. DETAILED DESCRIPTION OF THE INVENTION Definitions
[0062] For the purpose of interpreting this specification, the following definitions will be applied and whenever appropriate, terms used in the singular will also be included in the plural and vice versa.
[0063] As used here, "C1-6 alkyl" indicates an alkyl radical having from 1 to 6, in some cases from 1 to 4 carbon atoms, the radicals being linear or branched with a single or multiple branches. For example, "C1-6 alkyl" includes, but is not limited to, n-butyl, sec-butyl, isobutyl, tert-butyl; propyl, such as n-propyl, 2-methylpropyl or isopropyl; ethyl or methyl.
[0064] As used herein, the term "alkylene" refers to the divalent alkyl group as defined here above having 1 to 4 carbon atoms. Representative examples of alkylene include, but are not limited to, methylene, ethylene, n-propylene, iso-propylene, n-butylene, sec-butylene, iso-butylene, tert-butylene, and the like.
[0065] As used here, "C3-7 cycloalkyl" refers to saturated or partially unsaturated monocyclic, bicyclic, bridged or spirocyclic groups of 3 to 7 carbon ring atoms. Exemplary mocyclic hydrocarbon groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl and cyclohexenyl and the like.
[0066] As used herein, "halogen" or "halo" refers to fluorine, chlorine, bromine, and iodine; and more particularly, fluorine or chlorine.
[0067] As used herein, "halo C1-6 alkyl" or "C1-6 alkyl substituted by halo" refers to an alkyl radical, as defined above that is replaced by one or more halo radicals, as defined above, and it is particularly fluorine C1-6 alkyl, more particularly trifluoromethyl.
[0068] As used here, a "stereoisomer" refers to a compound made up of the same atoms linked by the same bonds, but having different three-dimensional structures, which are not interchangeable. The present invention contemplates several stereoisomers and mixtures thereof and includes "enantiomers", which refers to two stereoisomers whose molecules are mirror images that do not overlap each other.
[0069] As used herein, the term "amino acid conjugate" refers to conjugates of the compound of Formula I, II, III, IV or V with any suitable amino acid. Preferably, such suitable amino acid conjugates of the compounds of Formula I, II, III, IV or V will have the additional advantage of enhanced integrity in bile or intestinal fluids. Suitable amino acids include, but are not limited to, glycine, taurine and acyl glucuronide. Thus, the present invention encompasses glycine, taurine and acyl glucuronide conjugated to the compound of Formula I, II, III, IV or V.
[0070] As used herein, the term "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (eg, antibacterial agents, antifungal agents), isotonic agents, absorption, salts, preservatives, drugs, drug stabilizers, binders, excipients, disintegrating agents, lubricants, purifying agents, flavoring agents, dyes, and the like and combinations thereof, as is known to those skilled in the art (see, for example) example, Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329). Except to the extent that any conventional vehicle is incompatible with the active ingredient, its use in therapeutic or pharmaceutical compositions is contemplated.
[0071] As used herein, the term "therapeutically effective amount" refers to an amount of the compound of formula (I) that is sufficient to achieve the indicated effect. Therefore, a therapeutically effective amount of a compound of formula (I) used in the treatment of an FXR-mediated condition will be a sufficient amount for the treatment of the FXR-mediated condition.
[0072] As used here, the term "individual" refers to an animal. Typically the animal is a mammal. An individual also refers to, for example, primates (for example, humans, male or female), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds and the like . In certain modalities, the individual is a primate. In still other modalities, the individual is a human.
[0073] As used herein, the term "treat", "treating" or "treatment" of any disease or disorder refers in one embodiment to improving the disease or disorder (for example, slowing or arresting or reducing the development of disease or at least one of its clinical symptoms). In another modality "treating", "treating" or "treatment" refers to relieving or improving at least one physical parameter including those that cannot be discernible by the patient. In yet another modality, "treating", "treating" or "treatment" refers to modulating the disease or disorder, either physically, (for example, stabilization of a discernible symptom), physiologically, (for example, stabilization of a parameter physical), or both. In yet another embodiment, "treating", "treating" or "treatment" refers to preventing or delaying the onset or development or progression of the disease or disorder.
[0074] As used here, an individual is "in need of" treatment if that individual benefits biologically, medically or in the quality of life from such treatment.
[0075] As used here, the term "dyslipidemia" refers to an abnormality in, or abnormal amounts of, lipids and lipoproteins in the blood and the resulting disease states, caused by, exacerbated by, or in addition to, such abnormality (see, Dorland's Illustrated Medical Dictionary, 29th edition or later versions thereof, WB Saunders Publishing Company, New York, NY). Disease states covered within the definition of dyslipidemia as used here include hyperlipidemia, hypertriglyceridemia, low plasma HDL, high plasma LDL, high plasma VLDL, liver cholestasis, and hypercholesterolemia.
[0076] As used here, the phrase "diseases related to dyslipidemia" refers to diseases including but not limited to atherosclerosis, thrombosis, coronary artery disease, stroke, and hypertension. Diseases related to dyslipidemia also include metabolic diseases such as obesity, diabetes, insulin resistance, and complications thereof.
[0077] As used here, the term "cholestasis" refers to any condition in which the flow of bile from the liver is blocked, and can be intraepathic (for example, occurring within the liver) or extraepathic (for example occurring outside the liver).
[0078] As used herein, "liver fibrosis" includes liver fibrosis due to any cause, including, but not limited to, virus-induced liver fibrosis such as those due to hepatitis B and C; exposure to alcohol (alcoholic liver disease), pharmaceutical compounds, oxidative stress, cancer radiation therapy or industrial chemicals; and diseases such as primary biliary cirrhosis, adipose liver, obesity, non-alcoholic steatohepatitis, cystic fibrosis, hematochromatosis, and autoimmune hepatitis.
[0079] As used here, "FXR agonists" refers to an agent that binds directly and positively regulates FXR activity.
[0080] As used herein, the term "one, one (a)," "one, one (an)," "o, a (the)" and similar terms used in the context of the present invention (especially in the context of the claims ) must be interpreted to cover both the singular and the plural, unless otherwise indicated here or clearly contradicted by the context. Any definition here can be used in combination with any other definition to describe a structural group.
[0081] By convention, the drag element of such a definition is that which connects to the original portion. For example, the complex alkoxyalkyl group would represent an alkoxy group attached to the parent molecule through an alkyl group. Ways of Carrying Out the Invention
[0082] The present invention relates to compositions and methods for FXR. Various embodiments of the invention are described here. It will be recognized that features specified in each modality can be combined with other features specified to provide other modalities.
[0083] In a first embodiment, the compounds of the present invention are defined by Formula (I):

[0084] or a pharmaceutically acceptable salt thereof, wherein,
[0085] R0 is Ring A or C1-6 alkyl;
[0086] Ring A is aryl; 5 to 10 membered heteroaryl comprising 1 to 3 N, O or S heteroatoms; 4- to 6-membered heterocycle comprising 1 to 2 N, O or S heteroatoms; or C3-7 cycloalkyl; and said Ring A is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[0087] Ring B is aryl; 5 to 10 membered heteroaryl comprising 1 to 3 N, O or S heteroatoms; 4- to 6-membered heterocycle comprising 1 to 2 N, O or S heteroatoms; or C3-7 cycloalkyl; and said Ring B is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[0088] X is - (CR4R5) - or –C (O) -;
[0089] Y is –O–, - (CR4R5) -, * –O (CR4R5) - or –NR–, where "*" indicates the point of attachment of Y to the ring containing Z ring atoms;
[0090] Z1, Z2, Z3, and Z4 are each independently -CR3- or -N-;
[0091] L1 is * 1- (CR4R5) 1-2- or * 1- (CR4R5) -C (O) -NR-, where "* 1" indicates the point of attachment from L1 to N;
[0092] L2 is * 2- (CR4R5) 1-2-, * 2- (CR4R5) -C (O) -, * 2- (CR4R5) - C (O) -NR-, * 2- (CR4R5) 2-O -, * 2- (CR4R5) 2-NR-, * 2- (CR4R5) 2-SO2-, * 2- (CR4R5) 2-NR-C (O) -, or * 2- (CR4R5) -C (O) -NR- (CR4R5) -; where "* 2" indicates the connection point from L2 to N;
[0093] L3 is - (CR4R5) - or -C (O) -;
[0094] each R2 is independently halo, hydroxyl, C1-6 alkyl, or C1-6 alkyl substituted by halo;
[0095] each R3 is independently hydrogen, halo, or C1-6 alkyl; and
[0096] R, R4 and R5 are independently hydrogen or C1-6 alkyl.
[0097] In a second embodiment, the compounds of the present invention are defined by Formula (I) as defined in the first embodiment, in which
[0098] R0 is Ring A or C1-6 alkyl;
[0099] Ring A is aryl; 5 to 10 membered heteroaryl comprising 1 to 3 N, O or S heteroatoms; or C3-7 cycloalkyl; and said Ring A is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[00100] Ring B is aryl; 5 to 10 membered heteroaryl comprising 1 to 3 N, O or S heteroatoms; or C3-7 cycloalkyl; and said Ring B is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[00101] X is - (CR4R5) -;
[00102] Y is -O-, - (CR4R5) -, or * -O (CR4R5) -, where "*" indicates the point of attachment of Y to the ring containing the ring atoms Z;
[00103] Z1, Z2, Z3, and Z4 are each independently -CR3- or -N-;
[00104] L1 is * 1- (CR4R5) 1-2- where "* 1" indicates the point of attachment from L1 to N;
[00105] L2 is * 2- (CR4R5) 1-2-, * 2- (CR4R5) -C (O) -NR-, * 2- (CR4R5) 2-O-, * 2- (CR4R5) 2- NR- or * 2- (CR4R5) -C (O) -NR- (CR4R5) -; where "* 2" indicates the connection point from L2 to N;
[00106] L3 is -C (O) -;
[00107] each R2 is independently halo, C1-6 alkyl, or C1-6 alkyl substituted by halo;
[00108] each R3 is independently hydrogen, halo, or C1-6 alkyl;
[00109] R, R4 and R5 are independently hydrogen or C1-6 alkyl.
[00110] In a third embodiment, the compounds of the present invention are defined by Formula (II):

[00111] or a pharmaceutically acceptable salt thereof; on what,
[00112] R0 is Ring A or C1-6 alkyl; wherein Ring A is phenyl, pyridyl or cyclopropyl, each of which is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[00113] Ring B is selected from phenyl, pyridyl, 1H-indolyl, and C3-7 cycloalkyl, each of which is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[00114] X is - (CR4R5) -;
[00115] Y is -O-, - (CR4R5) - or * -O (CR4R5) -, where "*" indicates the point of attachment of Y to the ring containing Z ring atoms;
[00116] Z1, Z2, Z3, and Z4 are each independently -CR3- or -N-;
[00117] L1 is * 1- (CR4R5) 1-2- where "* 1" indicates the connection point from L1 to N;
[00118] L2 is * 2- (CR4R5) 1-2-, * 2- (CR4R5) -C (O) -NR-, * 2- (CR4R5) 2-O-, * 2- (CR4R5) 2- NR- or * 2- (CR4R5) -C (O) -NR- (CR4R5) -; where "* 2" indicates the connection point from L2 to N;
[00119] each R2 is independently halo, C1-6 alkyl, or C1-6 alkyl substituted by halo;
[00120] each R3 is independently hydrogen, halo, or C1-6 alkyl; and
[00121] R, R4 and R5 are independently hydrogen or C1-6 alkyl.
[00122] In a fourth mode, R0 in any of the above modes is selected from * 3-CH2C (CH3) 2-, * 3-CH2CH (CH3) -, and * 3-cyclopropane-1,1, -diyl , where "* 3" indicates the connection point from R0 to L2.
[00123] As a reference to any of the above modalities, Y can be selected from -O-, -CH2-, -C (CH3) 2-, and * -O-CH2-, where "*" indicates the point of bonding Y to the six-membered ring containing the ring atoms Z.
[00124] With reference to any of the above modalities, L1 can be - (CR4R5) - or -CH2-. In a variation, L1 is - (CR4R5) -.
[00125] With reference to any of the above modalities, L2 can be selected from - (CH2) -, * 2-CH2C (O) NH--, * 2- CH (CH3) C (O) NH-, * 2 -CH2C (O) NHCH2-, * 2- (CH2) 2O-, and * 2- (CH2) 2NH, where "* 2" indicates the point of attachment from L2 to N.
[00126] With reference to any of the above modalities, each R2 when present can be independently selected from halo, methyl, and trifluoromethyl.
[00127] In a fifth embodiment, the compounds of the present invention are represented by Formula (III):

[00128] or a pharmaceutically acceptable salt thereof; on what
[00129] Ring A is phenyl or pyridyl, each of which is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[00130] Ring B is selected from phenyl, pyridyl, 1H-indolyl, and cyclopentyl, each of which is substituted or unsubstituted by 1 to 2 substituents independently represented by R2;
[00131] L1 is - (CR4R5) -
[00132] L2 is selected from - (CH2) -, * 2-CH2C (O) NH-, * 2- CH (CH3) C (O) NH-, * 2-CH2C (O) NHCH2—, * 2- (CH2) 2O—, and * 2- (CH2) 2NH—; where "* 2" indicates the connection point from L2 to N;
[00133] X is CH2;
[00134] Y is selected from -O-, -CH2-, -C (CH3) 2- and * -O-CH2-, where "*" indicates the point of attachment of Y to the ring containing the ring atoms Z ;
[00135] Z1 is CR3 or N;
[00136] Z2 is CR3;
[00137] Z3 is CR3;
[00138] Z4 is CR3 or N;
[00139] each R2 is independently selected from halo, methyl, and trifluoromethyl;
[00140] each R3 is independently hydrogen, halo, or C1-6 alkyl; and
[00141] each of R4 and R5 is independently hydrogen or methyl.
[00142] In a sixth modality, Y in any of the above modalities is -O-. In an alternative embodiment, Y is -CH2-. In another alternative embodiment, Y is * -O-CH2-, where "*" indicates the point of attachment of Y to the ring containing the ring atoms Z.
[00143] In a seventh modality, L1 in any of the above modalities is -CH2-.
[00144] In an eighth mode, L2 in any of the above modes is selected from - (CH2) -, * 2-CH2C (O) NH-, * 2- (CH2) 2O-, and * 2- ( CH2) 2NH-; where "* 2" indicates the connection point from L2 to N.
[00145] In an eighth modality, L2 in any of the above modes is - (CH2) -.
[00146] In a tenth modality, R2 when present in any of the above modalities is independently fluorine or methyl. In a variation, each R2 is fluorine. In another variation, each R2 is methyl.
[00147] In an eleventh mode, R3 in any of the above modes is selected from hydrogen, fluorine, chlorine, and methyl.
[00148] In a twelfth mode, with reference to any of the above modes, Z1 is selected from CH, CF, CCH3, and N; Z2 is selected from CH, CF, CCl, and CCH3; Z3 is selected from CH, CF, CCl, and CCH3; and Z4 is CH or N.
[00149] With reference to any of the above modalities, each of the R, R4, R5 and R6 when present can independently be hydrogen or methyl. In a variation, R, R4, R5 and R6 when present are hydrogen.
[00150] In a thirteenth embodiment, the compounds of the present invention are represented by Formula (IV):

[00151] each phenyl ring is optionally substituted again by 1 to 2 substituents independently represented by R2, wherein R2 is fluorine or methyl;
[00152] L2 is selected from -CH2-, * 2-CH2CH2NH—, * 2-CH2CH2O-, and * 2-CH2C (O) NH-, where "* 2" indicates the point of attachment from L2 to N;
[00153] Z2 is selected from CH, CF, CCl, and CCH3; and
[00154] Z3 is selected from CH, CF, CCl, and CCH3.
[00155] In one mode, L2 in any of the above modes is - (CH2) -. In an alternative embodiment, L2 is * 2- (CH2) 2NH-. In another alternative embodiment, L2 is * 2-CH2C (O) NH-. In each mode, "* 2" indicates the connection point from L2 to N.
[00156] Particular compounds according to the present invention include, but are not limited to: 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4- dihydrochromene [4,3-c] pyrazol-3-carboxamido) ace-tamido) benzoic; 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid ; 3- (2- (8-chloro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid; 4-fluoro-3- (2- (1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (N- (2-fluorobenzyl) -1,8-dimethyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (N- (2-fluorobenzyl) -1,6-dimethyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-fluoro-N- (2-fluorobenzyl) - 1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (7-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (7-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (6,8-difluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1,7-dimethyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1,6-dimethyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-6-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid ; 4-fluoro-3- (2- (N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (7,8-difluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4- fluorobenzoic; 3- (2- (7,8-difluoro-N- (3-fluorobenzyl) -1- methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4- fluorobenzoic; 3- (2- (N-benzyl-7,8-difluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid; 4-fluoro-3- (2- (8-fluoro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid ; 4-fluoro-3- (2- (N- (2-fluorobenzyl) -1-methyl-4,5-dihydro-1H-pyrazolo [4,3-h] quinoline-3-carboxamido) acetamido) benzoic acid ; 4-fluoro-3- (2- (N- (2-fluorobenzyl) - 1-methyl-4,5-dihydro-1H-pyrazolo [3,4-f] quinoline-3-carboxamido) acid ) benzoic; (S) -4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido acid ) propanamido) benzoic; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl -4,5-dihydro-1H-benzo [g] indazole -3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-7-fluoro- N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid ; 4-fluoro-3- (2- (N- (3-fluorobenzyl) -1,5,5-trimethyl-4,5-dihydro-1H-benzo [g] indazol-3-carboxamido) acetamido) benzoic acid ; 4-fluoro-3- (2- (N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (3-fluorobenzyl) - 1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid; 3- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid; 3- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid; 3- (2- (N-benzyl-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid; 3- (2- (9-chloro-N- (2-fluorobenzyl) -1-methyl-4,5-dihydro-1H-benzo [2,3] oxepine [4,5-c] pyrazole-3 acid - carboxamido) acetamido) -4-fluorobenzoic; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (cyclopentylmethyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid; 3- (2 - (8-chloro-N- (cyclopentylmethyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -5-fluorobenzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-methylbenzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -2-methylpropanoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1- methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -2,2- dimethylpropanoic; 1 - ((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) methyl) cyclopropane- carboxylic; 4- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorobenzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic acid; 4- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic acid; N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-methylbenzoic acid; 4- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5- dimethylbenzoic; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -4-fluorobenzoic acid; 4- (2- (8-chloro-N- (cyclopentylmethyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5-dimethylbenzoic acid; 4- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5- difluorobenzoic; 4- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3- (trifluoromethyl) acid ) benzoic; 4- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5-difluorobenzoic acid; 3,5-difluoro-4- (2- (8-fluoro- N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy acid ) benzoic; 4- (2- (N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5-difluorobenzoic acid; 4- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorobenzoic acid; 4- (2- (N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorobenzoic acid; 4- (2- (N-benzyl-7,8-difluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorobenzoic acid; 3-fluoro-4- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic acid ; 4- (2- (7,8-difluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy acid -3- fluorobenzoic; 4- (2- (8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorobenzoic acid; 3-fluoro-4- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic acid ; 4- (2- (7,8-difluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy acid -3- fluorobenzoic; 3 - ((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) - 1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid; 3 - (((2- (8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino acid -4 fluorobenzoic; 3 - ((2- (8-chloro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino acid -4 fluorobenzoic; 4-fluoro-3 - ((2- (8-fluoro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl acid) amino) benzoic; 3 - ((2- (7,8-difluoro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino acid) -4- fluorobenzoic; 4-fluoro-3 - (((2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic; 4-fluoro-3 - ((2- (N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic acid; 3 - ((2- (N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-fluorobenzoic acid; 3 - ((2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-fluorobenzoic acid; 3 - ((2- (7,8-difluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino acid) -4-fluorobenzoic; 3 - ((2- (N-benzyl-7,8-difluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-fluorobenzoic acid ; 4- ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - (((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorobenzoic acid; 4 - ((8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-chloro-N- (3,5-difluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorobenzoic acid; 4 - ((8-chloro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-chloro-N- (2,3-difluorobenzyl) -1- methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-chloro-N- (3-fluoro-5-methylbenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid ; 4 - ((N- (3,5-difluorobenzyl) -8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-chloro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorobenzoic acid; 3-fluoro-4 - ((8-fluoro-1-methyl-N - (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - (((N - (((1H-indol-5-yl) methyl) -8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acid) benzoic; 5 - ((8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) methyl) picolinic acid; 4 - ((8-chloro-N - (((5-fluoropyridin-3-yl) methyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acid) benzoic; 4 - ((8-chloro-N - (((5-chloropyridin-3-yl) methyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acid) benzoic; 4 - ((8-chloro-N- (3-fluorobenzyl) -1-methyl -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -2-fluorobenzoic acid; 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -2-fluorobenzoic acid; N-benzyl-N- (4-carbamoylbenzyl) -8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamide; 4 - ((8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-chloro-N- (2-fluorobenzyl) - 1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorobenzoic acid; 3-fluoro-4 - ((8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid ; 4 - ((8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorobenzoic acid; 4 - ((8-chloro-N- (2-fluorobenzyl) -1- methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -2-fluorobenzoic acid; 6 - ((8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) nicotinic acid; 5 - ((8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -6-methylpicolinic acid; 4-fluoro-3 - (((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic; and 3 - (((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) - 4-fluorobenzoic; or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
[00157] More particular examples of the compounds according to the present invention include, but are not limited to: 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1 acid , 4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoyl; 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid ; 3- (2- (7-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (7,8-difluoro-N- (2-fluorobenzyl) -1- methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4- fluorobenzoic; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic acid; 3 - ((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic acid; 3 - ((2- (8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino acid -4 fluorobenzoic; 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; and 4-fluoro-3 - (((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl ) amino) benzoic; or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
[00158] In one embodiment, the present invention provides a compound according to any of the above embodiments, wherein the compound is in the form of a pharmaceutically acceptable salt selected from TRIS (2-amino-2-hydroxymethyl-1, 3-propanediol), arginine, lysine, sodium and meglumine salt.
[00159] The present invention also provides a compound of Formula (V):

[00160] wherein each phenyl ring is substituted or unsubstituted by 1 to 2 substituents independently represented by R2, wherein R2 is fluorine or methyl;
[00161] L2 is selected from * 2-CH2-, * 2-CH2CH2NH-, * 2-CH2CH2O-, and * 2-CH2C (O) NH-, where "* 2" indicates the connection point from L2 to N;
[00162] Z2 is selected from CH, CF, CCl, and CCH3;
[00163] Z3 is selected from CH, CF, CCl, and CCH3; and
[00164] R6 is C1-6 alkyl.
[00165] In another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound according to any of the above embodiments and variations or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.
[00166] In yet another aspect, the present invention provides a combination comprising a therapeutically effective amount of a compound according to any of the above embodiments and variations or a pharmaceutically acceptable salt thereof, and a second therapeutic agent.
[00167] In yet another aspect, the invention provides a method for treating a condition mediated by farnesoid X receptors (FXR) in an individual suffering from, comprising administering to the individual a therapeutically effective amount of a compound of any one. the above modalities and variations of a pharmaceutically acceptable salt thereof; and optionally in combination with a second therapeutic agent.
[00168] In yet another aspect, the invention provides a compound according to any of the above modalities and variations or a pharmaceutically acceptable salt thereof, and optionally in combination with a second therapeutic agent, for use in the treatment of a condition mediated by FXR.
[00169] In yet another aspect, the invention provides for the use of a compound of any of the above modalities and variations or a pharmaceutically acceptable salt thereof, and optionally in combination with a second therapeutic agent, for the preparation of a medicament for the treatment of an FXR-mediated condition in an individual.
[00170] In one embodiment, the condition mediated by FXR with respect to any of the above methods, uses or combinations, is a liver disease or a gastrointestinal disease. For example, the compounds of the invention can be used to treat FXR-mediated liver disease, in which liver disease is selected from cholestasis (e.g., intrahepatic cholestasis, estrogen-induced cholestasis, drug-induced cholestasis, pregnancy cholestasis, cholestasis associated with parenteral nutrition, progressive family cholestasis (PFIC)); Alagille syndrome, primary biliary cirrhosis (PBC), primary sclerosing cholangitis, ductopenic liver transplant rejection, graft versus host disease associated with liver transplantation, cystic fibrosis kidney disease, non-alcoholic adipose liver disease (NAFLD) , non-alcoholic steatohepatitis (NASH), alcoholic liver disease, and liver disease associated with parenteral nutrition. The compounds of the invention can also be used to treat an FXR-mediated gastrointestinal disease, in which the gastroin-testinal disease is selected from malabsorption of bile acids (including primary biliary acid diarrhea and secondary bile acid diarrhea), gastritis bile reflux, and inflammatory bowel disease such as Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, amusement colitis, indeterminate colitis and Behçet's disease.
[00171] More particularly, the condition mediated by FXR is non-alcoholic adipose liver disease (NAFLD) or non-alcoholic steatoepatitis (NASH). With reference to the combination of therapies of the invention, the other therapeutic agent may also be useful in the treatment of non-alcoholic adipose liver disease (NAFLD) or non-alcoholic steatoepatitis (NASH).
[00172] In one embodiment, the compounds of the invention are administered enterically; and more particularly, orally.
[00173] Unless otherwise specified, the term "compounds of the present invention" refers to compounds of Formula I, II, III, IV or V, pharmaceutically acceptable salt thereof, prodrugs, and inherently formed moieties (e.g. polymorphs, solvates and / or hydrates). The compounds of the present invention can be stereoisomers (including diastereoisomers and enantiomers), and can be a mixture of stereoisomers or a single stereoisomer. The compounds of the present invention can also be isotopically labeled tautomers and compounds (including deuterium substitutions). Other compounds of the invention are detailed in the Examples, infra.
[00174] Some of the compounds described here contain one or more asymmetric centers or axes and can thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined, in terms of absolute stereochemistry, as (R) or (S). The present invention is intended to include all possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures. Optically active (R) and (S) isomers can be prepared using chiral syntones or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituent can have an E or Z configuration. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent can have a cis or trans configuration. All tautomeric forms are also intended to be included.
[00175] Any formula given here is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have structures depicted by the formulas given here except that one or more atoms are replaced by an atom having an atomic mass or selected mass number. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2H, 3 H 11C 13C 14C 15N 18F 31P 32P 35S 36Cl 125I r ti m nt AH ,,,, N, F, P, P,, l and I respectively. The invention includes several isotopically labeled compounds as defined herein, for example, those in which radioactive isotopes, such as 3H, 13C, and 14C, are present. Such isotopically labeled compounds are useful in metabolic studies (with 14C), kinetic reaction studies (with, for example 2H or 3H), detection and imaging techniques, such as positron emission tomography (PET) or computed tomography by single photon emission (SPECT) including substrate or drug tissue distribution tests, or in the radioactive treatment of patients. In particular, an 18F or labeled compound may be particularly desirable for PET or SPECT studies. Isotopically labeled compounds of this invention and prodrugs thereof can generally be prepared by performing procedures disclosed in the schemes or in the examples and preparations described below by replacing a readily available isotopically labeled reagent with a non-isotopically labeled reagent.
[00176] In addition, substitution with heavier isotopes, particularly deuterium (eg 2H or D) may provide certain therapeutic advantages resulting from increased metabolic stability, for example, increasing half-life in vivo or reducing dosage requirements or a improvement in the therapeutic index. It is understood that deuterium in this context is considered to be a substitute for a compound of Formula (I). The concentration of such heavier isotopes, specifically, deuterium, can be defined by the isotopic enrichment factor. The term "isotopic enrichment factor" as used here means the relationship between isotopic abundance and the natural abundance of a specific isotope. If a substituent on a compound of this invention is denoted deuterium, that compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation in each designated deuterium atom) at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation) deuterium), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).
[00177] Isotopically labeled compounds of Formula I, II, III, IV and V can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described. Examples and accompanying Processes using an appropriate isotopically labeled reagent in place of the previously unlabeled reagent used.
[00178] Pharmaceutically acceptable solvates according to the invention include those in which the crystallization solvent can be isotopically substituted, for example, D2O, d6-acetone, d6-DMSO.
[00179] Compounds of the invention, for example, compounds of Formula I, II, III, IV and V that contain groups capable of acting as donors and / or acceptors for hydrogen bonds may be able to form crystals with suitable cocrystal builders. These co-crystals can be prepared from compounds of Formula I, II, III, IV or V by known cocrystalline procedures. Such procedures include grinding, heating, co-sublimation, co-fusion, or contacting in solution compounds of Formula I, II, III, IV or V with the cocrystal former under conditions of crystallization and isolating cocrystals thus formed. Suitable cocrystal builders include those described in WO 2004/078163. Accordingly, the invention also provides co-crystals comprising a compound of Formula I, II, III, IV or V.
Any asymmetric atom (for example, carbon or the like) of the compound (s) of the present invention can be present in racemic or enantiomerically enriched configuration, for example to (R), (S) or (R, S). In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R) or (S) configuration. Substituents in atoms with unsaturated bonds can, if possible, be present in cis (Z) or trans (E) form.
[00181] Therefore, as used here, a compound of the present invention can be in the form of one of the possible isomers, rotamers, atropisomers, tautomers or mixtures thereof, for example, as substantially pure geometric (cis or trans) isomers, di-astereomers, optical isomers (antipodes), racemates or mixtures thereof. Any mixtures resulting from isomers can be separated based on the physical-chemical differences of the constituents, in the pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and / or fractional crystallization. Any racemates resulting from the final product or intermediates can be resolved in optical antipodes by known methods, for example, by separating the diastereomeric salts of the same, obtained with an optically active acid or base, and releasing the optically active acidic or basic compound. In particular, the basic portion can thus be employed to resolve the compounds of the present invention into their optical antipodes, for example, by fractional crystallization of a salt former with an optically active acid, for example, tartaric acid, acid dibenzoyl tartaric acid, diacetyl tartaric acid, di-O tartaric acid, O'-p-toluoyl, mandelic acid, malic acid or camphor-10-sulfonic acid. Racemic products can also be resolved by chiral chromatography, for example, high pressure liquid chromatography (HPLC) using a chiral absorber. Pharmacology and Utility
[00182] The compounds of Formula I, II, III, IV and V in free form or in salt form, exhibit valuable pharmacological properties, for example, FXR modulation properties, for example, as indicated in in vitro tests and / or in vivo as provided for in the following sections, and are therefore indicated for therapy in the treatment of a disorder that can be treated by FXR modulation, such as those described below.
[00183] With development of the first synthetic FXR linker GW4064 as an instrument compound (Maloney et al., J. Med. Chem. 2000, 43 (16), 2971-2974; Willson et al., Med. Res. Rev. 2001, 21 (6) 513-22), and the development of an artificial and semi-synthetic bile acid ligand 6-alpha-ethyl-CDCA, the effects of FXR over-stimulation by potent agonists can be analyzed. Both ligands have been shown to induce bile flow in the attached bile duct of animals. In addition to choleretic effects, hepatoprotective effects can also be demonstrated (Pellicciari et al., J. Med. Chem. 2002, 45 (17), 3569-3572; Liu et al., J. Clin. Invest. 2003, 112 ( 11), 16781687). This hepatoprotective effect was again reduced to an antifibrotic effect that results from the suppression of Tissue Inhibitors of Matrix Metalloproteinases, TIMP-1 and 2, the induction of collagen deposit resolving Metalloproteinase Matrix 2 (MMP-2) in hepatic and stellate cells the subsequent reduction of alpha-collagen mRNA and transforming growth factor beta mRNA (TGF-beta) which are both pro-fibrotic factors by FXR agonists (Fiorucci et al., Gastroenterology 2004, 127 (5), 1497-1512; Fiorucci et al., Pharmacol. Exp. Ther. 2005, 314 (2), 584-595).
[00184] The antifibrotic activity of FXR is at least partially mediated by the induction of PPARY, another nuclear receptor, with which the antifibrotic activity is associated (Fiorucci et al., J. Pharmacol. Exp. Ther. 2005, 315 (1 ), 58-68; Galli et al., Gastroenterology 2002, 122 (7), 1924-1940; Pineda Torra et al., Mol. Endocrinol. 2003, 17 (2), 259-272). In addition, anti-cholestatic activity has been demonstrated in the linked biliary duct of animal models as well as in animal models of estrogen-induced cholestasis (Fiorucci et al., J. Pharmacol. Exp. Ther. 2005, 313 (2), 604 -612).
[00185] Genetic studies demonstrate that in hereditary forms of cholestasis (intrahepatic Progressive Familial Cholestasis = PFIC, Type I to IV), any nuclear location of the FXR itself is reduced as a consequence of a mutation in the FIC1 gene (in PFIC Type I , also called Byler's disease) (Chen et al., Gastroente-rology. 2004, 126 (3), 756-64; Alvarez et al., Hum. Mol. Genet. 2004; 13 (20), 2451-60 ) or target FXR gene levels encoding MDR-3 phospholipid export pump are reduced (in PFIC Type III). Taken together, there is a growing body of evidence that FXR binding compounds will demonstrate substantial clinical utility in the therapeutic regimen of chronic cholestatic conditions such as primary biliary cirrhosis (PBC) or primary sclerosing cholangitis (PSC) (reviewed in: Rizzo et al ., Curr. Drug Targets Immune Endocr. Metabol. Disord. 2005, 5 (3), 289-303; Zollner, MoI. Pharm. 2006, 3 (3), 231-51, Cai et al., Expert Opin. Ther Targets 2006, 10 (3), 409-421).
[00186] In addition, FXR appears to be involved in the regulation of many different physiological processes that are relevant in the etiology and for the treatment of diseases as diverse as cholesterol gallstones, metabolic disorders such as Type II Diabetes, dyslipidemia or obesity, chronic inflammatory diseases such as inflammatory bowel diseases or chronic intraepathic forms of cholestasis and many other diseases (Claudel et al., Arterioscler. Thromb. Vase. Biol. 2005, 25 (10), 2020-2030; Westin et al. , Mini Rev. Med. Chem. 2005, 5 (8), 719-727).
[00187] FXR has been shown to be a key regulator of serum triglycerides (Maloney et al., J. Med. Chem. 2000, 43 (16), 2971-2974; Willson et al., Med. Res. Rev. 2001 , 21 (6), 513-22). Recent reports indicate that the activation of FXR by synthetic agonists leads to a significant reduction in serum triglycerides, mainly in the reduced VLDL form, but also to reduce total serum cholesterol (Figge et al., J. Biol. Chem. 2004, 279 (4) , 2790-2799; Bilz et al., Am. J. Physiol. Endocrinol. Metab. 2006, 290 (4), E716-22). However, the reduction in serum triglycerides is not an independent effect. Treatment of db / db or ob / ob mice with synthetic FXR GW4064 agonists resulted in marked and combined reduction in serum triglycerides, total cholesterol, free fatty acids, and ketone bodies such as 3-OH Butyrate. In addition, activation of FXR interacts with the series of intracellular insulin signaling reactions in hepatocytes, resulting in a reduced glucose reduction of hepatic gluconeogenesis, but concomitantly with an increase in hepatic glycogen. Insulin sensitivity and glucose tolerance were positively impacted by FXR treatment (Stayrook et al., Endocrinology 2005, 146 (3), 984-91; Zhang et al., Proc. Natl. Acad. Sci. USA 2006 , 103 (4), 1006-1011; Cariou et al., J. Biol. Chem. 2006, 281, 1103911049; Ma et al., J. Clin. Invest. 2006, 116 (4), 1102-1109; Duran - Sandoval et al., Biochimie 2005, 87 (1), 93-98).
[00188] The compounds of the invention are also useful for the treatment of gastrointestinal diseases, including but not limited to malabsorption of bile acid (including primary bile acid diarrhea and secondary bile acid diarrhea), bile reflux gastritis and heart disease. inflammatory bowel (IBD). Malabsorption of bile acid, which leads to the excretion of excessive fecal bile acid and diarrhea in patients, is characterized by a cycle in which the regulation of bile acid synthesis feedback is interrupted, resulting in the production of additional bile acid. The regulation of bile acid synthesis feedback is under the control of a series of endocrine reactions, in which the activation of the nuclear bile acid receptor FXR induces the enteric expression of fibroblast growth factor 15 (FGF15) in rodents or FGF19 in humans. In the liver, FGF15 or FGF19 act together with FXR-mediated expression of a small heterodimer partner to suppress bile acid synthesis (Jung et al., Journal of Lipid Research 48: 2693-2700 (2007) Walters JR, Nat Rev Gastroenterol Hepatol 11 (7): 426-34 (2014)).
[00189] In another embodiment, the compounds according to the invention are useful for beneficially altering lipid profiles, including but not limited to a reduction in total cholesterol levels, a reduction in LDL cholesterol levels, a reduction in VLDL cholesterol levels , elevation of HDL cholesterol levels, and / or reduction of triglyceride levels. Thus, the present invention provides a method for treating conditions mediated by FXR such as dyslipidemia and diseases related to dyslipidemia comprising administering a therapeutically effective amount of a compound of the present invention to an individual in need thereof.
[00190] In another embodiment, the pharmaceutical compound or composition is used to treat a disease selected from the group consisting of disorders of lipids and lipoproteins such as hypercholesterolemia, hypertriglyceridemia, and atherosclerosis as a clinically manifested condition. which can be improved by the beneficial effects of FXR's in increasing HDL cholesterol, reducing serum triglycerides, increasing the conversion of cholesterol in the liver to bile acids and increased clearance and metabolic conversion of VLDL and other lipoproteins in the liver.
[00191] In another embodiment, the aforementioned compound and pharmaceutical composition are used for the preparation of a combined lipid-lowering drug, anti-cholestatic and anti-fibrotic effects of FXR target drugs can be exploited for the treatment of liver steatosis and associated syndromes such as non-alcoholic steatoepatitis (NASH), or for the treatment of cholestatic and fibrotic effects that are associated with alcohol-induced cirrhosis, or with virus-transmitted forms of hepatitis.
[00192] FXR appears to be involved in the control of antibacterial defense in the intestine (lnagaki et al., Proc. Natl. Acad. Sci. US A. 2006, 103 (10), 3920- 3905), and may have a beneficial impact on therapy for Inflammatory Bowel Disorders (IBD), particularly those forms where the upper (ileal) part of the intestine is affected (for example, ileal Crohn's disease) because this appears to be the site of action for controlling FXR over bacterial growth. In IBD, desensitization of the adaptive immune response is somewhat impaired in the intestinal immune system. Bacterial overgrowth can then be the trigger for the establishment of a chronic inflammatory response. Therefore, the dampening of bacterial growth by mechanisms transmitted by FXR can be an essential mechanism to prevent acute inflammatory episodes. Thus, the invention also relates to a compound according to formula (I) or a pharmaceutical composition comprising said compound to treat a disease related to inflammatory bowel diseases, such as Crohn's disease or ulcerative colitis. Restoration of intestinal barrier function and reduction in FXR-mediated non-commensal bacterial load is believed to be useful in reducing the exposure of bacterial antigens to the intestinal immune system and may therefore reduce inflammatory responses.
[00193] The invention also relates to a compound or pharmaceutical composition for the treatment of obesity and related diseases such as metabolic syndrome (combined conditions of dyslipidemia, diabetes and abnormally high body mass index), which can be overcome by reduction of serum triglycerides mediated by FXR, blood glucose and increased sensitivity to insulin and weight loss mediated by FXR. The compounds or pharmaceutical composition of the present invention are also useful in the preparation of a medicament to treat clinical complications of type I and type II diabetes, such as diabetic nephropathy, diabetic retinopathy, and Occlusive Peripheral Arterial Disease (PAOD).
[00194] In addition, conditions and diseases that result from chronic fibrotic and adipose organ degeneration due to the accumulation of lipid and specifically forced triglyceride and subsequent activation of pro-fibrotic reaction series can also be treated by applying the compounds or pharmaceutical composition of the present invention. Such conditions and diseases include non-alcoholic steatoepatitis (NASH) and chronic cholestatic conditions in the liver, diabetic glomerulosclerosis and nephropathy in the kidney, macular degeneration and diabetic retinopathy in the eye and neurodegenerative diseases such as Alzheimer's disease in the brain or diabetic neuropathies in the nervous system peripheral.
[00195] Examples of other FXR-mediated diseases include drug-induced bile duct injury, bile duct obstruction, gallstones, cholelithiasis, liver fibrosis, liver cirrhosis, alcohol-induced cirrhosis, dyslipidemia, atherosclerosis, diabetes, diabetic nephropathy, colitis , neonatal jaundice, prevention of severe neonatal jaundice, vein-occlusive disease, portal hypertension, metabolic syndrome, hypercholesterolemia, intestinal bacterial overgrowth, erectile dysfunction, and other conditions mediated by FXR leading to extraepathic cholestasis. Pharmaceutical Administration and Compositions
[00196] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable carrier. The pharmaceutical composition can be formulated for particular administration routines, such as oral administration, parenteral administration, and rectal administration, etc. In addition, the pharmaceutical compositions of the present invention can be made in a solid form (including without limitation capsules, tablets, pills, granules, powders or suppositories), or in a liquid form (including solutions without limitations, suspensions or emulsions). The pharmaceutical compositions may be subjected to conventional pharmaceutical operations, such as sterilization and / or may contain conventional inert diluents, lubricating agents, or buffering agents, as well as adjuvants, such as preservatives, stabilizers, wetting agents, emulsifiers and buffers, etc.
[00197] In a particular embodiment, the pharmaceutical composition is formulated for oral administration. Typically, pharmaceutical compositions are tablets or gelatin capsules comprising the active ingredient together with a) diluents, for example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine; b) lubricants, for example, silica, talc, stearic acid, its magnesium or calcium salt and / or polyethylene glycol; for pills too; c) binders, for example, magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carbonoxymethylcellulose and / or polyvinylpyrrolidone; if desired; d) disintegrants, for example, starches, agar, alginic acid or its sodium salt, or effervescent mixtures; and / or e) absorbents, dyes, flavors and sweeteners.
[00198] Tablets can be film coated or enteric coated according to methods known in the art.
[00199] Compositions suitable for oral administration include an effective amount of a compound of the invention in the form of tablets, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use are prepared according to any of the methods versed in the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets can contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients are, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example, starch, gelatin or acacia; and lubricating agents, for example, magnesium stearate, stearic acid or talc. The tablets are coated or uncoated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thus provide sustained action over a longer period. For example, a retardant material, such as glyceryl monostearate or glyceryl distearate can be employed. Formulations for oral use can be presented as rigid gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oily medium, for example, peanut oil, liquid paraffin or olive oil.
[00200] Certain injectable compositions are aqueous isotonic solutions or suspensions, and suppositories are advantageously prepared from emulsions or fatty suspensions. Said compositions can be sterilized and / or contain adjuvants, such as preservatives, stabilizers, humectants or emulsifiers, solution promoters, salts for regulating osmotic pressure and / or buffers. In addition, they may also contain other therapeutically valuable substances. Said compositions are prepared according to conventional mixing, granulation or coating methods, respectively, and contain about 0.1 to 75%, or contain about 1 to 50%, of the active ingredient.
[00201] Compositions suitable for transdermal application include an effective amount of a compound of the invention with a suitable vehicle. Suitable vehicles for transdermal delivery include pharmacologically acceptable absorbable solvents to assist passage through the host's skin. For example, transdermal devices are in the form of a bandage comprising a reinforcement member, a reservoir containing the compound optionally with vehicles, optionally a rate control barrier to release the compound from the host's skin at a controlled and predetermined rate over a period time, and features to attach the device to the skin.
[00202] Compositions suitable for topical application, for example, to the skin and eyes, include aqueous solutions, suspensions, ointments, creams, gels or sprayable formulations, for example, for aerosol release or the like. Such topical delivery systems will, in particular, be suitable for dermal application, for example, for the treatment of skin cancer, for example, for prophylactic use in sun creams, lotions, sprays and the like. They are, therefore, particularly suitable for topical use, including cosmetics, formulations well known in the art. Such may contain solubilizers, stabilizers, tonicity-enhancing agents, buffers and preservatives.
[00203] As used here, a topical application can also refer to inhalation or an intranasal application. They can conveniently be delivered in the form of a dry powder (or alone, as a mixture, for example, a dry mixture with lactose, or a mixed component particle, for example with phospholipids) from a dry powder inhaler or a presentation aerosol spray from a pressurized container, pump, spray, atomizer or nebulizer, with or without the use of a suitable propellant.
[00204] Dosage forms for topical or transdermal administration of a compound of this invention include powder, sprays, ointments, pastes, creams, lotions, gels, solutions, plasters and inhalers. The active compound can be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be desirable.
[00205] Ointments, pastes, creams and gels may contain, in addition to an active compound of this invention, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, traganthus, cellulose derivatives, polyethylene glycols, silicones , bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
[00206] Powders and sprays may contain, in addition to a compound of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powders, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.
[00207] Transdermal patches have the additional advantage of providing the controlled release of a compound of the present invention to the body. Such dosage forms can be made by dissolving or dispersing the compound in the appropriate medium. Absorption enhancers can also be used to increase the flow of the compound through the skin. The rate of such flow can be controlled either by providing a rate control membrane or by dispersing the active compound in a gel or polymer matrix.
[00208] Ointments, powders, ophthalmic solutions and the like, are also contemplated as included in the scope of this invention.
[00209] The present invention also provides anhydrous pharmaceutical compositions and dosage forms comprising the compounds of the present invention as active ingredients, as water can facilitate the degradation of certain compounds. Anhydrous pharmaceutical compositions and dosage forms of the invention can be prepared using anhydrous ingredients or containing low moisture content or low humidity content. An anhydrous pharmaceutical composition can be prepared and stored in such a way that its anhydrous nature is maintained. Therefore, anhydrous compositions are packaged using known materials to prevent exposure to water in such a way that they can be included in the appropriate formulation kits. Examples of suitable packaging include, but are not limited to, hermetically sealed thin sheets of metal, plastics, unit dose containers (eg vials), ampoule packs, and strip packs.
[00210] The invention also provides pharmaceutical compositions and dosage forms that comprise one or more agents that reduce the rate at which the compound of the present invention as an active ingredient will be broken down. Such agents, which are referred to herein as "stabilizers", include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers, etc.
[00211] The pharmaceutical composition or combination of the present invention can be in a single dose of about 1 to 1000 mg of active ingredient (s) for an individual of about 50 to 70 kg, or about 1 to 500 mg or about 1 to 250 mg, or about 1 to 150 mg, or about 0.5 to 100 mg, or about 1 to 50 mg of active ingredients. The therapeutically effective dosage of a compound, the pharmaceutical composition, or combinations thereof, is dependent on the individual's species, body weight, age and individual condition, the disorder or disease or severity of the same to be treated. A specialist doctor, clinician or veterinarian can easily determine the effective amount of each of the active ingredients needed to prevent, treat or inhibit the progress of the disorder or disease.
[00212] The dosage properties mentioned above are demonstrable in in vitro and in vivo tests using, advantageously, mammals, for example, mice, rats, dogs, monkeys or isolated organs, tissues and preparations thereof. The compounds of the present invention can be applied in vitro in the form of solutions, for example, aqueous solutions, and in vivo or enterically, parenterally, advantageously intravenously, for example, as a suspension or in aqueous solution. The in vitro dosage can vary in concentrations of about 10 to 3 molars and 10 to 9 molars. A therapeutically effective amount in vivo can vary depending on the means of administration, between about 0.1 to 500 mg / kg, or between about 1 to 100 mg / kg.
[00213] The compound of the present invention can be administered simultaneously with, or before, or after, one or more other therapeutic agent. The compound of the present invention can be administered separately, by the same or different route of administration, or together in the same pharmaceutical composition as the other agents.
[00214] In one embodiment, the invention provides a product comprising a compound of Formula I, II, III, IV or V, and at least one other therapeutic agent as a combined preparation for simultaneous, separate or sequential use in therapy. In one embodiment, therapy is the treatment of an FXR-mediated disease or condition. Products supplied as a combined preparation include a composition comprising a compound of Formula I, II, III, IV or V, and the other therapeutic agent (s) together in the same pharmaceutical composition, or the compound of Formula I, II, III, IV or V and the other therapeutic agent (s) separately, for example, in the form of a kit.
[00215] In one embodiment, the invention provides a pharmaceutical composition comprising a compound of Formula I, II, III, IV or V, and another therapeutic agent (s). It is contemplated that the invention provides a pharmaceutical composition comprising a compound of Formula I, II, III, IV or V in combination with naturally occurring non-toxic bile acid, such as ursodeoxycholic acid, as an aid in preventing possible vitamin depletion soluble in fat, secondary to treatment with FXR agonists. Therefore, the compounds of the invention can be administered concurrently with naturally occurring non-toxic bile acid, either as separate entities or as a single formulation comprising a compound of Formula I, II, III, IV or V, and naturally occurring bile acids .
[00216] Optionally, the pharmaceutical composition can comprise a pharmaceutically acceptable excipient, as described above.
[00217] In one embodiment, the invention provides a kit comprising two or more separate pharmaceutical compositions, at least one of which contains a compound of Formula I, II, III, IV or V. In one embodiment, the kit comprises means for retaining said compositions separately, such as a container, divided vial, or divided sheet metal package. An example of such a kit is an ampoule package, as typically used for packaging tablets, capsules and the like.
[00218] The kit of the invention can be used to administer different dosage forms, for example, oral and parenteral, to administer the separate compositions at different dosage intervals, or to titrate the separate compositions against each other. To assist compliance, the invention kit typically comprises directions for administration.
[00219] In the combination of therapies of the invention, the compound of the invention and the other therapeutic agent can be produced and / or formulated by the same or different productions. In addition, the compound of the invention and the other therapeutic can be combined into a combination therapy: (i) before the release of the combination product to doctors (for example, in the case of a kit comprising the compound of the invention and the other therapeutic agent); (ii) by the doctors themselves (or under the doctor's guidance) immediately before administration; (iii) in the patients themselves, for example, during the sequential administration of the compound of the invention and the other therapeutic agent.
Therefore, the invention provides for the use of a compound of Formula I, II, III, IV or V to treat an FXR-mediated disease or condition, in which the drug is prepared for administration, or administered with another therapeutic agent. The invention also provides a compound of Formula I, II, III, IV or V for use in a method of treating an FXR-mediated disease or condition, wherein the compound of Formula I, II, III, IV or V is prepared for administration, or administered with another therapeutic agent. The invention also provides another therapeutic agent for use in a method of treating an FXR-mediated disease or condition, wherein the other therapeutic agent is prepared for administration, or administered with, a compound of Formula I, II, III, IV or V.
[00221] The invention also provides for the use of a compound of Formula I, II, III, IV or V to treat an FXR-mediated disease or condition, in which the patient has previously (for example, within 24 hours) been treated with another therapeutic agent. Alternatively, the invention provides for the use of another therapeutic agent to treat an FXR-mediated disease or condition, in which the patient has previously (for example, within 24 hours) been with a compound of Formula I, II, III, IV or V. Additional Modalities
[00222] The present invention further encompasses additional embodiments described here.
[00223] Modality 1. A compound according to formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof,
on what,
[00224] R0 is Ring A or C1-6 alkyl; on what
[00225] Ring A is aryl; 5 to 10 membered heteroaryl comprising 1 to 3 N, O or S heteroatoms; 4- to 6-membered heterocycle comprising 1 to 2 N, O or S heteroatoms; or C3-7 cycloalkyl; each of which is substituted or unsubstituted by 1 to 2 substitutes each independently represented by R2; wherein L3 and R0 may be attached to the same or different ring atoms of Ring A; and C1-6 alkyl is optionally substituted by 1 to 2 C1-6 alkyl;
[00226] Ring B is aryl; 5 to 10 membered heteroaryl comprising 1 to 3 N, O or S heteroatoms; 4 to 6 membered heterocycle comprising 1 to 2 N, O or S heteroatoms; or C3-7 cycloalkyl; each of which is substituted or unsubstituted by 1 to 2 substituents each independently represented by R2;
[00227] X is (CR4R5) or C (O);
[00228] Y is O, (CR4R5), * O (CR4R5) or NR, where "*" indicates the point of attachment of Y to the ring containing the ring atoms Z;
[00229] Z1, Z2, Z3, and Z4 are each independently CR3 or N;
[00230] L1 is * 1- (CR4R5) 1-2- or * 1– (CR4R5) -C (O) -NR-, where "* 1" indicates the point of attachment from L1 to N;
[00231] L is - (cR R) 1-2-, - (cR R) -c (O), - (cR R) -c (O) -NR, * 2- (cR4R5) 2-O-, * 2- (cR4R5) 2-NR-, * 2- (cR4R5) 2-SO2-, * 2- (cR4R5) 2-NR- C (O) -, or * 2- (CR4R5) -C (O) -NR- (CR4R5); where "* 2" indicates the connection point from L2 to N;
[00232] L3 is - (CR4R5) - or -C (O) -;
[00233] each R2 is independently halo, hydroxyl, C1-6 alkyl, or C1-6 alkyl substituted by halo;
[00234] each R3 is independently hydrogen, halo, or C1-6 alkyl; and
[00235] R, R4 and R5 are independently hydrogen or C1-6 here.
[00236] Modality 2. A compound according to modality 1, or a salt, tautomer or stereoisomer thereof, in which the compound is represented by Formula (II),
on what
[00237] R0 is Ring A or C1-6 alkyl; on what
[00238] Ring A is selected from phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl and C3-7 cycloalkyl, each of which is substituted or unsubstituted by 1 to 3 substituents each independently represented by R2; wherein L3 and R0 may be attached to the same or different ring atoms of Ring A; and C1-6 alkyl is optionally substituted by 1 to 2 C1-6 alkyl;
[00239] Ring B is selected from phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1H-indolyl, and C3-7 cycloalkyl, each of which is substituted or unsubstituted by 1 to 2 substituents each independently represented by R2 ;
[00240] X is (CR4R5);
[00241] Y is O, (CR4R5) or * O (CR4R5), where "*" indicates the point of attachment of Y to the ring containing Z ring atoms;
[00242] Z1, Z2, Z3, and Z4 are each independently CR3 or N;
[00243] L1 is - (CR4R5) 1-2-,
[00244] L2 is * 2- (CR4R5) 1-2-, * 2- (CR4R5) -C (O) -NR-, * 2- (CR4R5) 2-O-, or * 2- (CR4R5) 2 -NR-; where "* 2" indicates the connection point from L2 to N; and
[00245] each R2 is independently halo, hydroxyl, C1-6 alkyl, or C1-6 alkyl substituted by halo;
[00246] each R3 is independently hydrogen, halo, or C1-6 alkyl;
[00247] R, R4 and R5 are independently hydrogen or C1-6 alkyl.
[00248] Mode 3. A compound according to mode 1 or 2, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, where R0 is selected from * 3-CH2C (CH3) 2-, * 3- CH2CH (CH3) -, and * 3-cyclopropane-1,1-di-yl-, where "* 3" indicates the point of attachment from R0 to L2.
[00249] Modality 4. A compound according to modality 1 or 2, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, in which the compound is represented by Formula (III):
on what
[00250] Ring A is phenyl or pyridyl, each of which is substituted or unsubstituted by 1 to 2 substituents each independently represented by R2;
[00251] Ring B is selected from phenyl, pyridyl, 1H-indolyl, and cyclopentyl, each of which is substituted or unsubstituted by 1 to 2 substituents each independently represented by R2;
[00252] L1 is - (CR4R5) -;
[00253] L2 is selected from * 2- (CH2) -, * 2-CH2C (O) NH-, * 2- CH (CH3) C (O) NH-, * 2-CH2C (O) NHCH2-, * 2- (CH2) 2O-, and * 2- (CH2) 2NH-; where "* 2" indicates the connection point from L2 to N;
[00254] X is CH2;
[00255] Y is selected from O, CH2, C (CH3) 2, and * O-CH2, where "*" indicates the point of attachment of Y to the ring containing the ring atoms Z;
[00256] Z1 is CR3 or N;
[00257] Z2 is CR3;
[00258] Z3 is CR3;
[00259] Z4 is CR3 or N; and
[00260] each R2 is independently selected from halo, methyl, and trifluoromethyl .;
[00261] each R3 is independently hydrogen, halo, or C1-6 alkyl; and
[00262] each of R4 and R5 is independently hydrogen or methyl.
[00263] Modality 5. A compound according to any one of modalities 1 to 4, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, where Y is O.
[00264] Modality 6. A compound according to any one of modalities 1 to 5, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, where L1 is -CH2-.
[00265] Modality 7. A compound according to any one of modalities 1 to 6, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, where L2 is selected from * 2- (CH2) -, * 2- CH2C (O) NH-, * 2- (CH2) 2O-, and * 2- (CH2) 2NH-; where "* 2" indicates the connection point from L2 to N.
[00266] Modality 8. A compound according to any one of modalities 1 to 6, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, where L2 is * 2- (CH2) -.
[00267] Modality 9. A compound according to any one of modalities 1 to 8, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein each R2 is independently fluorine or methyl.
[00268] Modality 10. A compound according to any one of modalities 1 to 9, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, in which each R3 is independently selected from hydrogen, fluorine, chlorine, and methyl.
[00269] Mode 11. A compound according to any one of modalities 1 to 9, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein
[00270] Z1 is selected from CH, CF, CCH3, and N;
[00271] Z2 is selected from CH, CF, CCl, and CCH3;
[00272] Z3 is selected from CH, CF, CCl, and CCH3; and
[00273] Z4 is CH or N.
[00274] Mode 12. The pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of modalities 1 to 11 and a pharmaceutically acceptable carrier.
[00275] Mode 13. A combination comprising a therapeutically effective amount of a compound according to any of modalities 1 to 11, and a second therapeutic agent being useful in the treatment of cholestasis, intraepathic cholestasis, estrogen-induced cholestasis, cholestasis drug-induced cholesterol, pregnancy cholestasis, cholestasis associated with parenteral nutrition, primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), progressive familial cholestasis (PFIC), non-alcoholic adipose liver disease (NAFLD), non-alcoholic steatopatitis (NASH), drug-induced bile duct injury, gallstones, liver cirrhosis, alcohol-induced cirrhosis, cystic fibrosis, bile duct obstruction, cholelithiasis, liver fibrosis, dyslipidemia, atherosclerosis, diabetes, diabetic nephropathy, colitis, neonatal jaundice, prevention of severe neonatal jaundice, veno-occlusive disease, portal hypertension, metabolic syndrome, hypercholesterolemia, bac overgrowth intestinal tract, or erectile dysfunction.
[00276] Mode 14. A method for treating a condition mediated by farnesoid X receptors (FXR) in an individual suffering from, comprising administering to the individual a therapeutically effective amount of a compound of any one of modalities 1 to 11, or a pharmaceutical composition thereof, and optionally in combination with a second therapeutic agent.
[00277] Modality 15. A pharmaceutical composition comprising a compound according to any one of modalities 1 to 11 for use in the treatment of an FXR-mediated condition.
[00278] Mode 16. A compound of Formula (VI)
on what:
[00279] Ring A is phenyl or pyridyl, each of which is substituted or unsubstituted by 1 to 2 substituents each independently represented by R2;
[00280] Ring B is selected from phenyl, pyridyl, 1H-indolyl, and cyclopentyl, each of which is substituted or unsubstituted by 1 to 2 substituents each independently represented by R2;
[00281] L1 is - (CR4R5) -;
[00282] L2 is selected from * 2- (CH2) -, * 2-CH2C (O) NH-, * 2- CH (CH3) C (O) NH-, * 2- (CH2) 2O-, and * 2- (CH2) 2NH-; where "* 2" indicates the connection point from L2 to N;
[00283] X is CH2;
[00284] Y is selected from O, CH2, C (CH3) 2, and * O-CH2, where "*" indicates the point of attachment of Y to the ring containing the ring atoms Z;
[00285] Z1 is CR3 or N;
[00286] Z2 is CR3;
[00287] Z3 is CR3;
[00288] Z4 is CR3 or N;
[00289] each R2 is independently selected from halo, methyl, and trifluoromethyl .;
[00290] each R3 is independently hydrogen, halo, or C1-6 alkyl;
[00291] each of R4 and R5 is independently hydrogen or methyl; and
[00292] R6 is C1-6 alkyl.
[00293] Mode 17. The compound according to mode 16, where Y is O.
[00294] Mode 18. The compound according to mode 16, where L1 is -CH2-.
[00295] Mode 19. The compound according to mode 16, where L2 is selected from * 2- (CH2) -, * 2-CH2C (O) NH-, * 2- (CH2) 2O-, and * 2- (CH2) 2NH-; where "* 2" indicates the connection point from L2 to N; and more particularly, where L2 is -CH2-.
[00296] Mode 20. The compound according to mode 16, in which each R2 is independently fluorine or methyl.
[00297] Mode 21. The compound according to mode 16, in which each R3 is independently selected from hydrogen, fluorine, chlorine, and methyl.
[00298] Modality 22. A method for preparing a crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [ 4,3-c] pyrazol-3-carboxamido) acetamido) benzoic, comprising the 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl) reaction step -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ace-tamido) benzoic with 2-amino-2-hydroxymethyl-propane-1,3-diol in a solvent such as a methanol solvent : dichloromethane (1: 1 by volume).
[00299] Mode 23. A crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3-c ] pyrazol-3-carboxamido) acetamido) benzoyl produced according to modality 22; and more particularly, wherein said crystalline form has a melting point of about 125 ° C as determined by differential scanning calorimetry.
[00300] Mode 24. A method for preparing a crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [ 4,3-c] pyrazol-3-carboxamido) acetamido) benzoic, comprising the step of: (i) reacting 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) acid - 1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoyl in a solvent such as methanol with aqueous L-arginine; and (ii) optionally, again crystallize the solid obtained from (i) in a solvent such as an acetonitrile: methanol solvent (2: 1 by volume).
[00301] Mode 25. A crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3-c ] pyrazole-3-carboxamido) acetamido) benzoyl produced according to modality 24; and more particularly, wherein said crystalline form has a melting point of about 206 ° C as determined by differential scanning calorimetry.
[00302] Mode 26. A method for preparing a crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [ 4,3-c] pyrazol-3-carboxamido) acetamido) benzoic, comprising the step of: (i) reacting 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) acid - 1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoyl in a solvent such as methanol with aqueous L-lysine; and (ii) optionally, again crystallize the solid obtained in (i) in a solvent such as acetonitrile.
[00303] Mode 27. A crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3-c ] pyrazol-3-carboxamido) acetamido) benzoyl produced according to modality 26.
[00304] Mode 28. A method for preparing a crystalline form of 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene) [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic, comprising the step of reacting 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) - 1- methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic with 2-amino-2-hydroxymethyl-propane-1,3-diol in a solvent such as methanol.
[00305] Mode 29. A crystalline form of 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3- c] pyrazol-3-carboxamido) ethyl) amino) benzoyl produced according to modality 28; and more particularly, wherein said crystalline form has a melting point of about 160 ° C as determined by differential scanning calorimetry.
[00306] Mode 30. A method for preparing a crystalline form of 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene) [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic, comprising the step of: (i) reacting 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl acid) ) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic and a solvent such as methanol with aqueous L-arginine; and (ii) optionally, crystallizing the solid obtained from (i) in a solvent such as acetonitrile.
[00307] Mode 31. A crystalline form of 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3- c] pyrazol-3-carboxamido) ethyl) amino) benzoyl produced according to modality 30; and more particularly, wherein said crystalline form has a melting point of about 161 ° C as determined by differential scanning calorimetry.
[00308] Mode 32. A method for preparing a crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [ 4,3-c] pyrazol-3-carboxamido) acetamido) benzoic, comprising the reaction step 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) - 1-methyl- 1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic in a solvent such as methanol with aqueous L-arginine.
[00309] Mode 33. A crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3-c ] pyrazol-3-carboxamido) acetamido) benzoyl produced according to modality 32; and more particularly, wherein said crystalline form has a melting point of about 206 oC as a given differential scanning calorimetry pot.
[00310] Mode 34. A method for preparing a crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [ 4,3-c] pyrazol-3-carboxamido) acetamido) benzoic, comprising the step of: (i) reacting 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) acid - 1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoyl in a solvent such as methanol with aqueous sodium hydroxide; and (ii) optionally, again crystallize the solid obtained in (i) in a solvent such as acetonitrile.
[00311] Mode 35. A crystalline form of 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3-c ] pyrazol-3-carboxamido) acetamido) benzoyl produced according to modality 34; and more particularly, wherein said crystalline form has a melting point of about 161 ° C as determined by differential scanning calorimetry.
[00312] Mode 36. A method for preparing a crystalline form of 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3- carboxamido) methyl) benzoic, comprising the step of reacting 4 - (((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acid) benzoyl with 2-amino-2-hydroxymethyl-propane-1,3-diol in a solvent such as methanol.
[00313] Mode 37. A crystalline form of 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid produced in accordance with modality 36; and more particularly, wherein said crystalline form has a melting point of about 195.6 oC as determined by differential scanning calorimetry.
[00314] Mode 38. A method for preparing a crystalline form of 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3- carboxamido) methyl) benzoic, comprising the step of: (i) reacting 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3- carboxamido) methyl) benzoyl in a solvent such as acetone with aqueous meglumine; and (ii) optionally, again comprising heating the solid obtained in (i) to a temperature ranging from 60 to 90 oC (for example, about 80 oC).
[00315] Mode 39. A crystalline form of 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid produced according to modality 38, step (i) having a dehydration point of about 71 oC as determined by differential scanning calorimetry; or obtained after another heating in step (ii) having a melting point of about 167.5 oC as determined by differential scanning calorimetry.
[00316] Mode 40. A method for preparing a crystalline form of 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3- carboxamido) methyl) benzoic, comprising the step of: (i) reacting 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3- carboxamido) methyl) benzoyl and meglumine in a solvent such as methanol; and (ii) again comprising heating the reagents to a temperature ranging from 60-90 oC.
[00317] Mode 41. A crystalline form of 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid produced in accordance with modality 40; and more particularly, in which the crystalline form has a melting point of about 180.6 oC as determined by differential scanning calorimetry. Processes for Manufacturing the Compounds of the Invention
[00318] The compounds of the present invention can be prepared by the routes described in the following schemes or in the Examples. All of the methods described here can be performed in any suitable order, unless otherwise indicated here or otherwise clearly contradicted by the context.
[00319] Compounds of Formula (I) in which L1 is methyl, L2 is * 2- CH2C (O) NH-, L3 is C (O) and R0 is Ring A can be prepared according to Scheme 1:
Layout 1
[00320] Compounds of Formula (I) in which L1 is methyl, L2 is * 2- CH2C (O) NH-, L3 is C (O) and R0 is Ring A can be prepared according to Scheme 2:
Scheme 2
[00321] Compounds of Formula (I) in which L1 is methyl, L2 is * 2- (CH2) 2NH-, L3 is C (O) and R0 is Ring A can be prepared according to Scheme 3:
Layout 3
[00322] Compounds of Formula (I) in which L1 is methyl, L2 is * 2- (CH2) 2O-, L3 is C (O) and R0 is Ring A can be prepared according to Scheme 4:
Caption: (aqueous) Scheme 4
[00323] Compounds of Formula (I) in which L1 is methyl, L2 is * 2- CH2C (O) NH-, L3 is C (O) and R0 is C1-6alkyl can be prepared according to Scheme 5:
Legend: oxalyl chloride Scheme 5
[00324] Compounds of Formula (I) in which L1 is methyl, L2 is * 2- (CH2) - C (O) NH (CH2), L3 is C (O) and R0 is Ring A can be prepared according to Scheme 6:
Layout 6
[00325] In each of the schemes above 1 to 6, X, Y, Z1, Z2, Z3, Z4, A and B are as defined in any of the above modalities. Generally, the tricyclic core (VI) is coupled to an amide side chain (VIIa, VIIb, VIIc, VIId or VIIe) with or without the use of a suitable amide coupling agent such as HATU; followed by hydrolysis to provide a compound of Formula (I).
[00326] In each of the Schemes above 1 to 5, the tricyclic core (VI) can be prepared according to Scheme 7 in which X, Y, Z1, Z2, Z3 and Z4 are as defined in any one of the above modalities. Typical conditions are exemplified in the synthesis of ethyl 2- (6-chloro-4-oxochroman-3-yl) -2-oxoacetate (I-1), 8-chloro-1-methyl-1,4-dihydrochromene [ 4,3-c] ethyl (I-3) pyrazol-3-carboxylate and 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxylic acid (I- 4), below.
Legend: diethyl oxalate Scheme 7
[00327] In Scheme 1 above, the amide side chain (VIIa) can be prepared according to Scheme 8, where A and B are as defined in any of the above embodiments. Typical conditions are exemplified in the synthesis of methyl 4-fluoro-3- (2 - ((3-fluorobenzyl) amino) acetamido) benzoate (I-16), infra.
Layout 8
[00328] In Scheme 2 above, the amide side chain (VIIb) can be prepared according to Scheme 9, where A and B are as defined in any of the above embodiments. Typical conditions are exemplified in the synthesis of methyl 4 - ((benzylamino) methyl) benzoate (I-23), infra.
Layout 9
[00329] In Scheme 3 above, the side chain of amide (VIIc) can be prepared according to Scheme 10, where A and B are as defined in any of the above modalities. Typical conditions are exemplified in the synthesis of 3 - ((2 - ((tert-butoxycarbonyl) amino) ethyl) amino) - methyl 4-fluorobenzoate (I-44), 3 - ((2-aminoethyl) amino) -4- methyl fluorobenzoate (I-45) and 4-fluoro-3 - ((2 - (((3-fluorobenzyl) amino) ethyl) amino) methyl (I-46) benzoate, infra.
Layout 10
[00330] In Scheme 4 above, the amide side chain (VIId) can be prepared according to Scheme 11, where B is as defined in any of the above modalities. Typical conditions are exemplified in the synthesis of 2 - ((2-fluorobenzyl) amino) ethanol (I-72), infra.
Layout 11
[00331] Each reaction step can be carried out in a manner known to those skilled in the art. For example, the reaction can be carried out in the presence of a suitable solvent or diluent or a mixture thereof. A reaction can also be carried out, if necessary, in the presence of an acid or base, with cooling or heating, for example, in the temperature range of approximately -30 oC to approximately 150 oC. In particular examples, a reaction is carried out in the temperature range of approximately 0 oC to 100 oC, and more particularly, in the temperature range of ambient temperature to approximately 80 oC, in an open or closed reaction vessel and / or in the atmosphere of an inert gas, for example, nitrogen.
[00332] The invention also relates to those forms of the process in which a compound obtainable as an intermediate at any stage of the process is used as a starting material and the remaining process steps are carried out, or in which a starting material is formed under reaction conditions or is used in the form of a derivative, for example, in a protected form or in the form of a salt, or a compound obtainable by the process according to the invention is produced under the process conditions and processed again in situ . Compounds of the invention and intermediates can be converted to one another according to methods generally known to those skilled in the art. Intermediates and final products can be worked and / or purified according to standard methods, for example, using chromatographic methods, distribution methods, (re) crystallization, and the like.
[00333] In the reactions described, reactive functional groups, for example, hydroxyl, amino, imino, thio or carboxy groups, where these are desired in the final product, can be protected to avoid their unwanted participation in the reactions. A characteristic of protecting groups is that they can be easily removed (for example, without the occurrence of unwanted side reactions), for example, by solvolysis, reduction, photolysis or alternatively under physiological conditions (for example, by enzymatic cleavage). Conventional protecting groups can be used according to standard practice (see, for example, T.W. Greene and P. G. M. Wuts in "Protective Groups in Organic Chemistry," 4th Ed., Wiley-Interscience, 2006, and subsequent versions thereof).
[00334] All the process steps mentioned above, mentioned here before and below can be carried out under reaction conditions that are known to those skilled in the art, including those mentioned specifically, in the absence of or, usually, in the presence of solvents or diluents , including, for example, solvents or diluents that are inert with respect to used reagents and dissolve them, in the absence or presence of catalytic, condensing or neutralizing agents, for example, ion exchangers, such as cation exchangers, for example example, in the form of H +, depending on the nature of the reaction and / or the reagents at reduced, normal, or elevated temperature, for example, in a temperature range of about -100 ° C to about 190 ° C, including, for example, from approximately -80 ° C to approximately 150 ° C, for example, at about -80 to -60 ° C, at room temperature, at about -20 to 40 ° C or at reflux temperature, under pressure atmospheric or in a fe chado, when appropriate under pressure, and / or in an inert atmosphere, for example, under an argon or nitrogen atmosphere.
[00335] At all stages of the reactions, mixtures of isomers that are formed can be separated into the individual isomers, for example, diastereoisomers or enantiomers, or into any desired mixtures of isomers, for example, racemates or mixtures of di-astereoisomers. Mixtures of obtainable isomers according to the invention can be separated in a manner known to those skilled in the art in the individual isomers; diastereoisomers can be separated, for example, by division between mixtures of polyphasic solvents, recrystallization and / or chromatographic separation, for example on silica gel or by, for example, medium pressure liquid chromatography on a reverse phase column, and racemates can be separated, for example, by the formation of salts with optically pure salt-forming reagents and separation of the mixture of diastereoisomers thus obtained, for example, by fractional crystallization, or by chromatography on optically active column materials.
[00336] The solvents from which those solvents that are suitable for any particular reaction can be selected include those specifically mentioned or, for example, water, esters, such as lower alkyl-lower alkanoates, for example, ethyl acetate, ethers , such as aliphatic ethers, for example, diethyl ether, or cyclic ethers, for example, tetrahydrofuran or dioxane, liquid aromatic hydrocarbons, such as benzene or toluene, alcohols, such as methanol, ethanol or 1- or 2 -propanol, nitriles, such as acetonitrile, halogenated hydrocarbons, such as methylene chloride or chloroform, acid amides, such as dimethylformamide or dimethyl acetamide, bases, such as heterocyclic or heteroatomic nitrogen bases, for example, pyridine or N-methylpyrrolidin-2-one, anhydrides of carboxylic acids, such as anhydrides of lower alkanoic acid, for example, acetic, cyclic anhydride, linear hydrocarbons or r amified, such as cyclohexane, hexane or isopen-tane, meticyclohexane, or mixtures of these solvents, for example, aqueous solutions, unless otherwise indicated in the process description. Such solvent mixtures can also be used in carrying out, for example, by chromatography or partitioning.
The compounds of the present invention are either obtained in free form, as a salt thereof, or as prodrug derivatives thereof. When both a basic group and an acid group are present in the same molecule, the compounds of the present invention can also form internal salts, for example, zuterether molecules. In many cases, the compounds of the present invention are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar to these. As used herein, the terms "salt" or "salts" refer to an acid addition or base addition salt of a compound of the invention. "Salts" include, in particular, "pharmaceutically acceptable salt". The term "pharmaceutically acceptable salts" refers to salts that retain the biological efficacy and properties of the compounds of this invention and, which are typically not biologically or otherwise undesirable.
[00338] Salts of compounds of the present invention having at least one salt-forming group can be prepared in a manner known to those skilled in the art. For example, salts of compounds of the present invention having acidic groups can be formed, for example, by treating the compounds with metal compounds, such as alkali metal, salts of desirable organic carboxylic acids, for example, the sodium salt of 2-ethylhexanoic acid , with an organic alkali metal or alkaline earth metal compounds, such as the corresponding hydrogen hydroxides, carbonates or carbonates, such as sodium or potassium hydroxide, hydrogen carbonate or carbonate, with corresponding calcium compounds or with ammonia or a suitable organic amine, stoichiometric amounts or just a small excess of the salt-forming agent preferably being used. Acid addition salts of compounds of the present invention are obtained in a customary manner, for example, by treating the compounds with a suitable acid or anion-exchange reagent. Internal salts of compounds of the present invention containing acid or basic salt-forming groups, for example, a free carboxy group and a free amino group, can be formed, for example, by the neutralizing salts, such as acid addition salts, to the point isoelectric, for example, with weak bases, or by treatment with ion exchangers. Salts can be converted to free compounds according to methods known to those skilled in the art. Metals and ammonia salts can be converted, for example, by treatment with acids, and suitable acid addition salts, for example, by treatment with a suitable basic agent.
[00339] Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids, for example, acetate, aspartate, benzoate, besylate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, camphorsulfonate, chloride salts / hydrochloride, chloroteophyllinate, citrate, ethanisulfonate, fumarate, gluceptate, gluconate, glucuronate, hypurate, iodate / iodide, isethionate, lactate, lactobionate, lauryl sulfate, malate, maleate, malonate, mandelaate, mesylate, methylate sulfate, na , nitrate, octa-decanoate, oleate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, polygalacturonate, propionate, stearate, succinate, sulfosalicylate, tartrate, tosylate and trifluoroacetate.
[00340] Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.
[00341] Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.
[00342] Inorganic bases from which salts can be derived include potassium, ammonia, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonia, potassium, sodium, calcium and magnesium salts.
[00343] Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Certain organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine and tromethamine.
[00344] Other pharmaceutically acceptable salts can be derived from L-arginine, TRIS (2-amino-2-hydroxymethyl-1,3-propanediol), adipic acid (adipate), L-ascorbic acid (ascorbate), capric acid (caprate ), sebacic acid (sebacate), 1-hydroxy-2-naphthoic acid (xinafoate), L-glutamic acid (glutamate), glutaric acid (glutarate), triphenylacetic acid (triphenatate) and galactic acid / mucic acid (mucate) .
[00345] The pharmaceutically acceptable salts of the present invention can be synthesized from a parent compound, a basic or acidic moiety, by conventional chemical methods. Generally, such salts can be prepared by reacting free acid forms of these compounds with a stoichiometric amount of the appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate or the like), or by reacting free forms of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are typically carried out in water or in an organic solvent, or in a mixture of the two. Generally, use of non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile is desirable, when feasible. Lists of additional suitable salts can be found, for example, in "Remington: The Science and Practice of Pharmacy," 21st Ed., Pharmaceutical Press 2011; and in "Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (2nd Edition of Rev., Wiley-VCH 2011, and subsequent versions thereof).
[00346] The present invention also provides prodrugs of the compounds of the present invention which converts in vivo to the compounds of the present invention. A prodrug is an active or inactive compound that is chemically modified through in vivo physiological action, such as hydrolysis, metabolism and the like, in a compound of this invention following administration of the prodrug in an individual. The suitability and techniques involved in the production and use of prodrugs are well known to those skilled in the art. Prodrugs can be conceptually divided into two non-exclusive categories, bioprecursor prodrugs and vehicle prodrugs. (See, The Practice of Medicinal Chemistry, Ch. 31-32 (Ed. Wermuth, Academic Press, San Diego, Calif., 2001, and subsequent versions thereof). activity compared to the corresponding active drug compound which contains one or more protecting groups and is converted to an active form by metabolism or solvolysis, either the active drug form or any released metabolic products must have acceptably low toxicity.
[00347] Vehicle prodrugs are composed of drugs that contain a transport portion, for example, that improve absorption and / or localized distribution to a site (s) of action. Desirably for such a vehicle prodrug, the link between the drug moiety and the transport moiety is a covalent bond, the prodrug is inactive or less active than the drug compound, and any release moiety of the transport moiety is acceptably non-toxic. For prodrugs where the transport portion is intended to increase absorption, typically the release of the transport portion should be rapid. In other cases, it is desirable to use a moiety that provides slow release, for example, certain polymers or other moieties, such as cyclodextrins. Vehicle prodrugs can, for example, be used to improve one or more of the following properties: increased lipophilicity, increased duration of pharmacological effects, increased site specificity, reduced toxicity and adverse reactions, and / or increased drug formulation (eg stability, solubility in water, suppression of an unwanted organoleptic or physicochemical property). For example, lipophilicity can be increased by esterifying (a) hydroxyl groups with lipophilic carboxylic acids (for example, a carboxylic acid having at least one lipophilic moiety), or (b) groups of carboxylic acids with lipophilic alcohols (for example , an alcohol having at least a lipophilic moiety, for example, aliphatic alcohols).
Exemplary prodrugs are, for example, esters of free carboxylic acids and S-acyl derived from thiols and O-acyl derived from alcohols or phenols, where acyl has a meaning as defined herein. Suitable prodrugs are constantly derived from pharmaceutically acceptable esters convertible by solvolysis under physiological conditions to the parent carboxylic acid, for example, alkyl esters, cycloalkyl esters, alkenyl esters, benzyl esters, mono- or di-substituted alkyl esters, such as the w- (amino, mono or di alkylamino, carboxy, alkoxycarbonyl) alkyl esters, the alkyl esters of a- (alkanoyloxy, alkoxycarbonyl or di alkylaminocarbonyl), such as the pivaloyloxymethyl ester and the like conventionally used in the art. In addition, the amines were masked as substituted arylcarbonyloxymethyl derivatives that were cleaved by esterases in vivo releasing the free drug and formaldehyde (Bundgaard, J. Med. Chem. 2503 (1989), and subsequent versions thereof). In addition, prodrugs containing an acidic NH group, such as imidazole, imide, indole and the like, have been masked as N-acyloxymethyl groups (Bundgaard, Design of Prodrugs, Elsevier (1985), and subsequent versions thereof). Hydroxy groups were masked as esters and ethers. EP 039,051 (Sloan and Little) describes Mannich-based hydroxamic acid prodrugs, their preparation and use.
[00349] Furthermore, the compounds of the present invention, including their salts, can also be obtained in the form of hydrates, or their crystals can, for example, include the solvent used for crystallization. Different crystalline forms may be present. The compounds of the present invention can inherently or by design form solvates with pharmaceutically acceptable solvents (including water); moreover, it is intended that the invention covers both solvated and unsolvated forms. The term "solvate" refers to a molecular complex of a compound of the present invention (including pharmaceutically acceptable salts thereof) with one or more solvent molecules. Such solvent molecules are those commonly used in the pharmaceutical technique, which are known to be harmless to the recipient, for example, water, ethanol, and the like. The term "hydrate" refers to the complex where the solvent molecule is water. The compounds of the present invention include salts, hydrates and solvates thereof, they can inherently or by design form polymorphs.
[00350] Compounds of the invention in non-oxidized form can be prepared from N-oxides of compounds of the invention by treatment with a reducing agent (for example, sulfur, sulfur dioxide, triphenyl phosphine, lithium borohydride, sodium borohydride, trichloride phosphorus, tribromide, or the like) in a suitable inert organic solvent (eg, acetonitrile, ethanol, aqueous dioxane, or the like) at 0 to 80 ° C. EXAMPLES
[00351] The examples provided here are offered to illustrate, but not to limit the compounds of the invention, as well as the preparation of such compounds and intermediates. It is understood that if there is a discrepancy between the name and the structure of a particular compound, the structure should be considered correct since the names of the compounds were generated from the structures. All variables are as defined here.
[00352] All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents and catalysts used to synthesize the compounds of the present invention are either commercially available or can be produced by methods of organic synthesis known to those versed in the technique (Houben-Weyl Science of Synthesis volumes 1-48, Georg Thieme Verlag, and subsequent versions thereof). In addition, the compounds of the present invention can be produced by methods of organic synthesis known to those skilled in the art as shown in the following examples.
[00353] Temperatures are given in degrees Celsius. If not mentioned otherwise, all evaporations are carried out under reduced pressure, typically between about 15 mm Hg and 100 mm Hg (20 to 133 mbar). The structure of final products, intermediates and starting materials is confirmed by standard analytical methods, for example, microanalysis and spectroscopic characteristics, for example, MS, IR, NMR. Abbreviations used are those conventional in the art.
[00354] Unless otherwise noted, melting points were calculated by differential scanning calorimetry (DSC) using the TA Q2000 or TA Discovery differential scanning calorimeters at a scan rate of 10 ° C / min. The accuracy of the sample temperature measurement is generally within approximately ± 1 oC.
[00355] NMR and LC-MS methodologies are well known in the art. The methods described here are illustrative only, and are not considered limiting.
[00356] NMR. NMR spectra were recorded on a Bruker AVANCE-400 spectrometer that operates at a proton frequency of 400.13 MHz equipped with a 5mm QNP cryoprobe (1H / 13C / 19F / 31P); or a Bruker AVANCE-600 spectrometer operating at a frequency of 600.13 MHz equipped with a 5 mm Z-gradient TCI cryoprobe or a 5 mm TXI cryoprobe. Unless otherwise indicated, samples were acquired at a temperature of 300 ° K, and spectra were referenced to the appropriate solvent peak.
[00357] LC-MS methods. Mass spectra were acquired in the LC-MS system using electrospray, chemical impact and electron ionization methods from a variety of instruments. Typical methods are described below. Method 1:
[00358] Waters Acquity Binary Gradient Pump; Waters Acquity PDA detector. Waters auto-sampler; Waters Quadratro micro API mass spectrometer with ESI and APCI ion source; UPLC column: Waters Acquity; BEH; C18 1.7 um 50x2.1 mm; Mobile Phase: (A) H2O + 0.025% TFA and (B) Acetonitrile + 0.025% TFA. Gradient: 0.4 mL / min, initial 15% B rise to 95% B for 3.0 minutes, then maintained up to 4.0 minutes, return to 15% B at 4.1 minutes until completion cycle, then equilibrate the column for 2.0 minutes. MS scan: 100 to 1000 amu in 0.5 seconds for one channel; Diode Array Detector: 200 nm and 400 nm. Method 2:
[00359] Waters Acquity Binary Gradient Pump; Waters Acquity PDA detector. Waters auto-sampler; Waters Quattro micro API Mass Spectrometer with ESI and APCI ion source; UPLC column: Acquity Wa-ters; BEH; C18 1.7 um 50x2.1 mm; Mobile Phase: (A) H2O + 0.025% TFA and (B) Acetonitrile + 0.025% TFA. Gradient: 0.4mL / min, initial 20% B rise to 90% B for 2.0 minutes, then maintained up to 4.0 minutes, return to 20% B at 4.1 minutes until end of the cycle, then balance the column for 2.0 minutes. MS scan: 100 to 1000 amu in 0.5 seconds for one channel; Diode Array Detector: 200 nm and 400 nm. Method 3:
[00360] Agilent 1200sl / 6140 system; UPLC column: Waters Acquisition; HSS T3; C18 1.8 um 50x2.0 mm; Mobile Phase: (A) H2O + 0.05% TFA and (B) Acetonitrile + 0.035% TFA. Gradient: 0.9mL / min, initial 10% B rise to 100% B for 1.95 minutes, then return to 10% B at 2.00 minutes until the end of the cycle. MS scan: 100 to 1000 amu in 0.5 seconds for one channel; Diode Array Detector: 190 nm and 400 nm; Drain tube temperature: 50 ° C and N 2 gas flow: 2.81 kg / cm2 (40 psi) for ELSD Detector. Method 4:
[00361] Agilent 1100sl / 1946 system; UPLC column: Waters Atlantis; C18 1.8 um 50x2.0 mm; Mobile Phase: (A) H2O + 0.05% TFA and (B) Acetonitrile + 0.035% TFA. Gradient: 1.0 mL / min, initial 10% B rise to 90% B for 3.00 minutes, then return to 10% B at 3.5 minutes until the end of the cycle. MS scan: 100 to 1000 amu in 0.5 seconds for one channel; Diode Array Detector: 190 nm and 400nm; Drain tube temperature: 50 ° C and N2 gas flow: 2.81 kg / cm2 (40 psi) for ELSD detector.
[00362] Analytical Method: WATERS ZQ SHIMADZU LEAP CTC, ZORBAX SB-C8 30 * 4.6 mm, 3.5 um, UV1: 220 nm, UV 2: 254 nm, A: H2O (0.03% TFA) , B: CH3CN (0.05% TFA), Flow: 2,000 (ml / min), Time /% B: 0/5, 1.90 / 95, 2.30 / 95, 2.31 / 5, 2.50 / 5. Abbreviations Boc butyl carboxy tertiary br broad d doublet doublet doublet DIAD diisopropyl diode azodicarboxylate DIEA diethylisopropylamine DMF N, N-dimethylformamide DMSO dimethylsulfoxide ESI electrospray ionization EtOAc ethyl acetate 2-HATH-hydroxy -il) - 1,1,3,3-tetramethyl uronium hr (s) HPLC hour (s) high performance liquid chromatography LCMS liquid chromatography and MeOH mass spectrometry methanol MS multiplet m mass spectrometry mg min (s) ml milligram minute (s) milliliter mmol millimol m / z mass-to-load ratio NaHDMS bis (trimethylsilyl) sodium amide NMM N-methylmorpholine NMR nuclear magnetic resonance PPh3 triphenylphosphine rt retention time s triplet singlet TFA trifluoroacetic acid THF tetra-hydrofuran TRF (2-amino-2-hydroxymethyl-1,3-propanediol) Tris ^ HCl aminotris (hydroxymethyl) methane hydrochloride Intermediates 2- (6-chloro-4-oxochroman-3-yl) -2-ethyl oxoacetate (I- 1)
Caption: diethyl oxalate
[00363] A solution of 6-chloro-2,3-dihydrochromen-4-one (109 mmol) in THF was treated with a solution of NaHMDS (60 mL, 120 mmol, 1.1 eq, 2M in THF) in THF at -78 oC under nitrogen. After stirring for 30 minutes, diethyl oxalate (22 mL, 163 mmol, 1.5 eq.) Was added at - 78 ° C dropwise and then stirred for 1 hour at room temperature. Subsequently the reaction was quenched with 1N HCl until the pH value was adjusted to 3. The resulting mixture was extracted with EtOAc (200 ml x 3). The combined organic phase was washed with brine and dried over Na2SO4. The solvent was removed in vacuo to provide I-1 as a yellow solid. MS (m / z): 283 (M + H) +. 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxylate (I-3)

[00364] To a heated solution of I-1 (109 mmol) in ethanol was added 1-methylhydrazine (109 mmol,) and the solution was stirred for 12 hours at room temperature. After the solvent was removed in vacuo, the residue was purified by column chromatography (petroleum ether: ethyl acetate = 95: 5) to provide I-2 and the desired product I-3 as a yellow solid. 1H-NMR: (300MHz, CDCl3): δ 7.69 (d, J = 2.7 Hz, 1H), 7.13 (dd, J = 2.4 Hz, J = 8.7 Hz, 1H), 6.86 (d, J = 8.7 Hz, 1H), 5.43 (s, 2H), 4.38 (q, J = 7.2 Hz, 2H), 4.22 (s, 3H), 1 , 41 (t, J = 7.2 Hz, 3H). MS (m / z): 293 (M + H) +. 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxylic acid

[00365] To a solution of I-3 (44 mmol) in 100 mL of THF / water (4: 1) was added NaOH (89 mmol, 2 eq.) And the mixture was heated at 60 oC for 8 hours. Subsequently, the resulting white solid was filtered and washed with methanol and the solid was treated with 100 ml of 1N HCl. The resulting solid was collected by filtration, washed with methanol completely and dried in vacuo to provide I-4 as a white solid. 1H-NMR (300MHz, CDCl3): δ 13.05 (brs, 1H), 7.72 (d, J = 2.7 Hz, 1H), 7.34 (dd, J = 2.7 Hz, J = 8.7 Hz, 1H), 7.06 (d, J = 8.7 Hz, 1H), 5.41 (s, 2H), 4.19 (s, 3H). MS (m / z): 265 (M + H) +.
[00366] The following intermediates were prepared according to the procedures described for synthesis of I-4 using the appropriate reagents.

4-fluoro-3- (2 - ((3-fluorobenzyl) amino) acetamido) methyl benzoate (I-16)

[00367] Bromoacetyl bromide (1.1 mL, 12.5 mmol) was added dropwise to a mixture of methyl 3-amino-4-fluorobenzoate (10.9 mmol) and diisopropylethylamine (2, 8 mL, 16.4 mmol) in dichloromethane at 0 oC. After stirring for 30 minutes at 0 oC, the reaction mixture was diluted with dichloromethane and water. The layers were separated and the aqueous phase was washed with dichloromethane (2x). The combined organic layers were dried over MgSO4 and concentrated (vacuum) to provide a dark brown liquid. LCMS showed that amide was the major component and aniline was a minor component. The mixture was dissolved in DMF (30 ml) and potassium carbonate (12.3 mmol) was added. To the mixture, 3-fluorobenzylamine (10.9 mmol) was added and the reaction stirred at room temperature for 16 hours. The reaction was diluted with water (300 ml) and the aqueous solution was extracted with ethyl acetate (3x). The combined organic layers were washed with water, brine, and were dried over MgSO4. The mixture was filtered and concentrated and the crude residue was purified by flash chromatography (silica gel, 0 to 60% ethyl acetate / hexanes) to provide I-16. 1H NMR (400 MHz, d4-MeOH): δ 8.80 (dd, J = 8.0, 4.0 Hz, 1H), 7.89-7.85 (m, 1H), 7.56-7 , 51 (m, 1H), 7.38-7.29 (m, 3H), 7.25 (app dt, J = 8.3, 6.0 Hz, 1H), 4.35 (s, 2H) , 4.09 (s, 2H), 3.91 (s, 3H).
[00368] The following intermediates were prepared according to the procedures described for the synthesis of I-16 using the appropriate reagents.

2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) acetic acid (I-22)
Caption: oxalyl chloride Step 1
[00369] To a suspension of 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxylic acid (I-4) (1.9 mmol) in CH2Cl2 (10 ml), catalytic amount of DMF (25 µl) and oxalyl chloride (0.78 ml, 8.9 mmol) were added dropwise at 0 ° C. The resulting suspension was warmed to room temperature and stirred for 1 hour. The solvent was removed under vacuum until complete dryness (it was necessary to remove oxalyl chloride completely). The obtained residue was dissolved with CH2Cl2 (10 mL) and then dropwise in a solution of methyl 2 - ((3-fluorobenzyl) amino) acetate (1.9 mmol) in CH2Cl2 (10 mL) in the presence of DIEA (0.56 mL, 3.8 mmol). The reaction mixture was stirred for 30 minutes at room temperature and was then charged with 50 ml of water. Organic layer was separated and the aqueous layer was extracted with CH2Cl2 (50 ml). Combined organic layers were washed with H2O and brine successively, dried over Na2SO4, filtered and concentrated to provide the crude product which was purified by column chromatography (0 to 60% EtOAc in hexane vs) to provide 2- ( 8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acetate (I-21). 1H NMR (400 MHz, CDCl3): δ 7.47 - 7.43 (m, 2H), 7.34 - 7.29 (m, 1H), 7.23 - 7.19 (m, 1H), 7 , 18 - 7.04 (m, 2H), 6.97 (dd, J = 2.3, 8.7 Hz, 1H), 5.57 - 4.12 (m, 9H), 3.77 - 3 , 74 (m, 3H). Mixture of rotamers. MS (m / z): 444.1 (M + H) +. Step 2
[00370] 2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acetate (I-21) (1.62 mmol) was dissolved in THF / MeOH / H2O (3: 2: 1, 10 mL) and followed by the addition of LiOH monohydrate (0.408 g, 9.72 mmol). The reaction mixture was stirred at room temperature for 1 hour and diluted with 10 ml of water and acidified to pH = 2.0, Solid was collected and dried to provide 2- (8-chloro-N- (2-fluorobenzyl) acid -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) acetic (I-22). 1H NMR (400 MHz, DMSO): δ 12.73 (s, 1H), 7.71 (t, J = 2.6 Hz, 1H), 7.42 - 7.30 (m, 3H), 7, 24 - 7.15 (m, 2H), 7.07 - 7.02 (m, 1H), 5.41 - 4.10 (m, 9H). Mixture of rotamers. MS (m / z) 430.0 (M + H) +. 4 - Methyl ((benzylamino) methyl) benzoate (I-23)

[00371] Benzyl amine (8.65 mL, 79 mmol) (Aldrich) and methyl 4-formylbenzoate (79 mmol) (Aldrich) were dissolved in methanol (Volume: 350 mL) and the mixture was stirred for 2 hours at temperature resulting in a white precipitate. The reaction mixture was cooled to 0 ° C and sodium boroidide (158 mmol) (Aldrich) was added in portions over 15 minutes after stirring for 1 hour at 0 ° C and the mixture was allowed to warm to room temperature. After stirring for an additional 1 hour the mixture was cooled in an ice bath and the reaction was slowly quenched with water (15 mL, 833 mmol). The resulting mixture was allowed to stir for 15 minutes. After removing the solvent (aspirator) the mixture was dissolved in EtOAc and washed with H2O. The aqueous phase was washed with EtOAc and the combined organic layers were dried (MgSO4). The crude material was purified by chromatography (silica gel, charged pure with DCM rhinse, 0 - 40% DCM / (10% MeOH in DCM)) to provide I-23. 1H NMR (400 MHz, DMSO): δ 9.17 (bs, 2H), 7.95-7.96 (m, 2H), 7.56 - 7.54 (m, 2H), 7.44 - 7 , 34 (m, 5H), 4.19 (s, 1H), 4.12 (s, 1H), 3.78 (s, 3H).
[00372] The following intermediates were prepared according to the procedures described for the synthesis of I-23 using the appropriate reagents.


3 - ((2 - (((tert-butoxycarbonyl) amino) ethyl) amino) -4-methyl fluorobenzoate (I-44)

[00373] Methyl 3-amino-4-fluorobenzoate (8.4 mmol) and tert-butyl (2-oxoethyl) carbamate (12.6 mmol) were dissolved in MeOH (100 mL) and AcOH (10 mL). After stirring for 1 hour at room temperature the reaction mixture was treated with NaCNBH3 (16.6 mmol) and stirred for 2 hours. After removing the solvent (aspirator) the mixture was dissolved in EtOAc and washed with NaHCO3 (saturated, 1x). The aqueous phase was washed with EtOAc (1x) and the combined organic layers were dried (MgSO4). The product (I -44) was made raw for the next step. 1H NMR (400 MHz, CDCl3): δ 7,407.38 (m, 2H), 7.02 (dd, J = 8.8 Hz, 11.1, 1H), 4.88 (s, 1H), 3, 91 (s, 3H), 3.40-3.38 (m, 4H), 1.47 (s, 9H). MS (m / z): 313.1 (M + H) +. Methyl 3 - ((2-aminoethyl) amino) -4-fluorobenzoate (I-45)

[00374] 3 - ((2 - ((tert-butoxycarbonyl) amino) ethyl) amino) -4-methyl fluorobenzoate (1.57 g) was dissolved in dichloromethane (10 ml) and treated with TFA (10 ml). The reaction stirred at room temperature for 1 hour. The solvent was azeotropically removed with dichloromethane and the cream colored solid (I-45) was used directly for the next step. 1H NMR (400 MHz, DMSO): δ 7.78 (s, 3H), 7.24 (m, 3H), 5.93 (s, 1H), 3.83 (s, 3H), 3.50 - 3.22 (m, 5H), 3.04 (s, 2H). MS (m / z): 213.1 (M + H) +. Methyl 4-fluoro-3 - ((2 - ((3-fluorobenzyl) amino) ethyl) amino) benzoate (I-46)

[00375] A mixture of methyl 3 - ((2-aminoethyl) amino) -4-fluorobenzoate (5.0 mmol), sodium bicarbonate (5.0 mmol) and methanol (30 mL) stirred at room temperature for 30 minutes. 3-fluoro-benzaldehyde (5.0 mmol) was added and stirred at room temperature for 2 hours. Sodium borohydride (7.4 mmol) was added in portions. The reaction stirred at room temperature for 30 minutes. LCMS showed that the reaction was complete. The solvent was removed and the residue was diluted with ethyl acetate (50 ml). The organic solution was washed with water and brine, dried over MgSO4, and concentrated. The crude material was purified by flash chromatography (silica 5 to 40% ethyl acetate / hexanes) to provide (I-46) as a white solid. 1H NMR (400 MHz, DMSO): δ 7.33 (td, J = 6.3, 8.1 Hz, 1H), 7.25 (dd, J = 1.9, 8.6 Hz, 1H), 7.23 - 7.10 (m, 4H), 7.08 - 6.98 (m, 1H), 5.67 (s, 1H), 3.82 (s, 3H), 3.74 (s, 2H), 3.20 (q, J = 6.2 Hz, 2H), 2.72 (t, J = 6.3 Hz, 2H). MS (m / z): 321.1 (M + H) +.
[00376] The following intermediates were prepared according to the procedures described for the synthesis of I-46 using the appropriate reagents.
Methyl 4- (2-aminoethoxy) -3-fluorobenzoate (I-51)

[00377] To a mixture of methyl 3-fluoro-4-hydroxybenzoate (17mmol), cesium carbonate (25mmol) and DMF (30ml) was added tert-butyl (2-bromoethyl) carbamate (20mmol). The resulting suspension is stirred at 60 ° C for 4 hours. The reaction was diluted with water (100 ml) and the resulting precipitate was collected by vacuum filtration. The filter cake was washed with water and dried in vacuo to provide pure I-50. I-50 (17 mmol) was taken up in dichloromethane (12 ml) and treated with TFA (5 ml). The reaction stirred at room temperature for 30 minutes. LCMS showed that the reaction was complete. The reaction was concentrated to dryness. The raw material was taken up in dichloromethane and stirred with solid sodium bicarbonate for 8 hours to remove any TFA residue. The mixture was filtered and the filtrate was dried to provide I-51 as a clear oil. 1H NMR (400 MHz, DMSO): δ 7.79 (d, J = 9.4 Hz, 1H), 7.73 (dd, J = 1.6, 11.8 Hz, 1H), 7.33 ( t, J = 8.6 Hz, 1H), 4.22 (t, J = 5.4 Hz, 2H), 3.84 (s, 3H), 3.12 (t, J = 5.4 Hz, 2H). Methyl 4- (2- (benzylamino) ethoxy) -3-fluorobenzoate (I-52)

[00378] A mixture of methyl 4- (2-aminoethoxy) -3-fluorobenzoate (I-51) (1.4 mmol) and sodium bicarbonate (1.4 mmol) in methanol (5 mL) was stirred at room temperature room for 30 minutes. Acetic acid (1.0 mL) and aldehyde was added and the reaction stirred at room temperature for 2 hours. NaCNBH3 (2.1 mmol) was added in three portions. The reaction stirred at room temperature for an additional 15 minutes. LCMS indicated that the reaction was complete. The reaction was diluted with water and extracted with ethyl acetate. The organic extracts were combined and washed with water and brine, dried over MgSO4 and concentrated to provide I-52. No further purification was necessary. MS (m / z): 304.1 (M + H) +.
[00379] The following intermediate was prepared according to the procedure described for the synthesis of I-52 using the appropriate reagents.
Methyl 4- (2 - ((tert-butoxycarbonyl) (2-fluorobenzyl) amino) ethoxy) -3- fluorobenzoate (I-55)

[00380] Di-tert-butyl dicarbonate (3.0 mmol) was added to a mixture of 2 - ((2-fluorobenzyl) amino) ethanol (3.0 mmol) in dichloromethane (25 mL) and NaOH ( 1 M in H2O, 9.6 mL, 9.6 mmol) at room temperature. The reaction mixture was stirred for 16 hours and then diluted with H2O (50 ml) and dichloromethane (25 ml). After separating the layers, the aqueous phase was washed with dichloromethane (2x, 50 ml) and the combined organic extracts were washed with H2O (1x 50 ml), dried (Na2SO4) and concentrated (vacuum cleaner). The crude mixture was purified by chromatography (silica gel, 0 to 60% EtO-Ac / hexanes) to provide I-54 as an oil. Diisopropyl azodicarboxylate (0.41 mL, 2.1 mmol) was added dropwise to a mixture of methyl I-54 (1.0 mmol), methyl 3-fluoro-4-hydroxybenzoate (2.0 mmol) and PPh3 (2.0 mmol) in THF (15 mL) at 0 ° C. After stirring for 10 minutes, the reaction mixture was allowed to warm to room temperature naturally, and stirred for an additional 4 hours. The reaction mixture was diluted with Et2O (50 ml) and H2O (50 ml) and the layers were separated. The aqueous phase was washed with Et2O (2x, 50 ml) and the combined organic extracts were dried (MgSO4) and then concentrated (vacuum). The crude mixture was purified by chromatography (silica gel, 0 to 25% EtOAc / hexanes) to provide an inseparable mixture of methyl I-55 and 3-fluoro-4-hydroxybenzoate. The mixture was concentrated, diluted with EtOAc (100 ml) and then washed with NaOH (x2, 1 M, 25 ml) to remove methyl 3-fluoro-4-hydroxybenzoate from the mixture. The organic phase was dried (MgSO4) and concentrated (vacuum) to provide I-55. 1H NMR (400 MHz, CDCI3): δ 7.69 - 7.55 (m, 2H), 7.23 - 7.06 (m, 2H), 7.00 - 6.86 (m, 2H), 6 , 86 - 6.71 (m, 1H), 4.56 - 4.45 (m, 2H), 4.14 - 3.99 (m, 2H), 3.75 (s, 3H), 3.59 - 3.44 (m, 2H), 1.40 - 1.25 (m, 9H). Methyl 3-fluoro-4- (2 - ((2-fluorobenzyl) amino) ethoxy) benzoate (I-56)

[00381] Trifluoroacetic acid (2 ml) was added to a solution of I-55 (0.9 mmol) in dichloromethane (8 ml) at room temperature. After stirring for 1 hour, the reaction mixture was concentrated (aspirator), diluted with MeOH and neutralized by passing the mixture through SPE-carbonate polymer binding cartridges (6 cartridges, units of 100 mg each). The solvent was removed (vacuum) to provide I-56. MS (m / z): 322.1 (M + H) +; rt = 0.981. Methyl 3- (2-aminoacetamido) -4-fluorobenzoate (I-58)

[00382] HATU (8.2 mmol) was added to a mixture of N-Boc glycine (6.8 mmol), methyl 3-amino-4-fluorobenzoate (6.2 mmol), diisopropylethylamine (3 , 6 ml, 20.5 mmol) and DMF (15 ml). The reaction was stirred at room temperature for 1 hour. LCMS indicated that the reaction was complete. The reaction was diluted with water and extracted with ethyl acetate. The organic extracts were dried over MgSO4 and concentrated. No further purification was necessary. The crude amide (I-57) (9.2 mmol) was dissolved in dichloromethane (15 ml) and treated with trifluoroacetic acid (8 ml). The reaction was stirred at room temperature for 1 hour. LCMS indicated that the reaction was complete. The reaction was concentrated to dryness and the raw material was purified by preparative HPLC to provide I-58. 1H NMR NMR (400 MHz, d4-MeOH) δ 8.71 (br d, J = 8.0 Hz, 1H), 7.79-7.75 (m, 1H), 7.21 (dd, J = 10.3, 8.2 Hz, 1H), 3.87 (s, 2H), 3.82 (s, 3H). MS (m / z): 227.1 (M + H) +. Methyl 4-fluoro-3- (2 - ((3-methylbenzyl) amino) acetamido) benzoate (I-59)

[00383] A mixture of methyl 3- (2-aminoacetamido) -4-fluorobenzoate (I-58) (1.5 mmol) and sodium bicarbonate (1.5 mmol) in methanol (5 mL) was stirred at room temperature room for 30 minutes. Acetic acid (1.0 mL) and 3-methylbenzaldehyde were added and the reaction stirred at room temperature for 2 hours. NaCNBH3 (2.2 mmol) was added in three portions. The reaction stirred at room temperature for an additional 15 minutes. LCMS indicated that the reaction was complete. The reaction was concentrated and purified by preparative HPLC to provide I-59. MS (m / z): 331.1 (M + H) +; r.t. = 1.51.
[00384] The following intermediate was prepared according to the procedure described for the synthesis of I-59 using the appropriate reagents.
(S) -3- (2 - ((tert-butoxycarbonyl) amino) propanamido) -4-methyl fluorobenzoate (I-60)

[00385] Isobutyl chloroformate (5.3 mmol) was added to a solution of N- (tert-butoxycarbonyl) -L-alanine (5.0 mmol) and N-methylmorpholine (5.3 mmol) in THF (25 mL ) at 0 ° C. After stirring for 30 minutes, methyl 3-amino-4-fluorobenzoate (5.3 mmol) was added as a solid and the resulting mixture was stirred for 16 hours at room temperature. After removing the solvent (aspirator), the residue was dissolved in EtOAc (50 ml) and washed with NaHCO3 (saturated, 50 ml), HCl (0.1 M in H2O, 50 ml) and brine (50 ml). The organics were dried (MgSO4) and concentrated (vacuum). The crude material was purified by chromatography (silica gel, 0 to 40% EtOAc / hexanes) to provide I-60, 1H NMR (600 MHz, CDCl3): δ 8.96 (dd, J = 2.0, 7, 6 Hz, 1H), 8.67 (br s, 1H), 7.83 - 7.75 (m, 1H), 7.17 - 7.11 (m, 1H), 4.94 (br s, 1H ), 4.36 (br s, 1H), 3.90 (s, 3H), 1.50 - 1.43 (m, 12H). (S) -4-fluoro-3- (2 - ((2-fluorobenzyl) amino) propanamido) methyl benzoate (I-62)

[00386] Trifluoroacetic acid (1.0 mL) was added to a solution of I-60 (0.9 mmol) in dichloromethane (5 mL) at room temperature. after stirring for 30 minutes, the solvent was removed (vacuum) and the residue was dissolved in dichloromethane (25 ml) and washed with NaHCO3 (saturated, 25 ml). The aqueous phase was washed with dichloromethane (2x, 25 ml) and the combined organic extracts were dried (Na2SO4) and concentrated (vacuum) to provide 190 mg of I-61 as an oil. 2-Fluorobenzaldehyde (94 mg, 0.8 mmol) was added to a solution of I-61 (0.8 mmol) in MeOH (9 mL) and AcOH (1 mL). After stirring for 1 hour at room temperature, the reaction mixture was treated with NaBH4 (1.6 mmol) and stirred for 30 minutes. After removing the solvent (aspirator), the mixture was dissolved in EtOAc (25 ml) and washed with NaHCO3 (saturated, 25 ml). The aqueous phase was washed with EtOAc (2x, 25 ml) and the combined organic extracts were dried (MgSO4) and concentrated (vacuum). The material was purified by chromatography (silica gel, 0 to 60% EtOAc / hexanes) to provide I-62. MS (m / z): 349.2 (M + H) +; rt = 1.097. 1-Methyl-4,5-dihydro-1H-benzo [2,3] oxepine [4,5-c] pyrazole-3-carboxylic acid (I-64)
Caption: steps
[00387] Intermediate I-64 was prepared according to the procedure described for the preparation of intermediate I-4, starting with 3,4-dihydrobenzo [b] oxepin-5 (2H) -one (I-63 , commercially available). 1H NMR (400MHz, DMSO): 1H NMR (400 MHz, DMSO): δ 12.77 (s, 1H), 7.80 (d, J = 2.6 Hz, 1H), 7.44 (dd, J = 8.6, 2.6 Hz, 1H), 7.22 (d, J = 8.7 Hz, 1H), 4.33 (t, J = 6.3 Hz, 2H), 4.05 (s , 3H), 3.12 (t, J = 6.3 Hz, 2H). MS (m / z): 279.0 / 281.0 (M + H) + (chlorine isotope standard); rt = 1.302. 2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) acetic acid (I-66)
Step 1
[00388] To a 20 mL reaction vessel, reagents were added in the following order: glycine (10 mmol), AcOH (2 mL), MeOH (12 mL), water (2 mL). This was stirred until complete dissolution. Then 2-fluorobenzaldehyde (5.0 mmol) was added. After 20 minutes, sodium cyanoborohydride (3.0 mmol) was added. The reaction was stirred 10 minutes, at which time an additional portion of sodium cyanoborohydride was added (3.0 mmol). The reaction was stirred for 10 minutes, filtered, and then purified by modified acetic acid (0.05%) reverse phase chromatography (10 to 50%, water-ACN) and subsequently recrystallized by MeOH / water 1: 5 (2 mL) to provide 2 - ((2-fluorobenzyl) amino) acetic acid (I-65) as a white powder. 1H NMR (400 MHz, DMSO): δ 7.61 (t, J = 8.1 Hz, 1H), 7.54 (app q, J = 7.8 Hz, 1H), 7.36-7.28 (m, 2H), 4.24 (s, 2H), 3.92 (s, 2H). MS (m / z): 184.1 (M + H) +. Step 2
[00389] To a 100 mL reaction vessel, reagents were added in the following order: 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxylic acid (I-4 ) (1.5 mmol), THF (15 mL), and N-methyl morpholine (NMM) (1 mL, 6.9 mmol). This was stirred until complete dissolution. Then 2-chloro-4,6-dimethoxy-1,3,5-triazine (1.6 mmol) was added and this solution was stirred for 40 minutes at 50 ° C until a completely white precipitate formed. The precipitate was physically stirred with rapid stirring to ensure that all solids were mixed well. Then 2- ((2-fluorobenzyl) amino) acetic acid (I-65) (2.3 mmol) was added and the reaction was stirred for 30 minutes at 50 ° C and then diluted with 5 ml of MeOH and in then purified by modified acetic acid (0.05%) reverse phase chromatography (30 to 80%, water / ACN) and subsequently recrystallized from MeOH / water 1: 1 (20 mL) to provide a white powder (I-66 ). 1H NMR (400 MHz, CDCl3): δ 10.05 (br s, 1H), 7.39 (app t, J = 7.8 Hz, 2H), 7.23-7.20 (m, 1H), 7.14 (d, J = 8.3 Hz, 1H), 7.09 (t, J = 8.2 Hz, 1H), 7.02 (t, J = 8.2 Hz, 1H), 6, 84 (d, J = 8.3 Hz, 2H), 5.44-5.40 (m, 2H), 4.82 (br s, 1H), 4.59 (br s, 1H), 4.05 -4.02 (m, 1H), 4.01 (s, 3H). MS (m / z): 430.1 / 432.1 (M + H) + (chlorine isotope standard). 2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) acetic acid (I-68)
Caption: oxalyl chloride Step 1
[00390] To a suspension of 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxylic acid (I-4) (1.9 mmol) in CH2Cl2 (10 ml), catalytic amount of DMF (25 µl) and oxalyl chloride (0.78 ml, 8.9 mmol) were added dropwise at 0 ° C. The resulting suspension was warmed to room temperature and stirred for 1 hour. The solvent was removed under vacuum until complete dryness (it was necessary to remove oxalyl chloride completely). The residue obtained was dissolved with CH2Cl2 (10 mL) and then dropped into a solution of methyl 2 - ((3-fluorobenzyl) amino) acetate (1.9 mmol) in CH2Cl2 (10 mL) in the presence of DIEA (0.56 mL, 3.8 mmol). The reaction mixture was stirred for 30 minutes at room temperature and then 50 ml of water was charged. Organic layer was separated and the aqueous layer was extracted with CH2Cl2 (50 ml). Combined organic layers were washed with H2O and brine successively, dried over Na2SO4, filtered and concentrated to provide the crude product, which was purified by column chromatography (0 to 60% EtOAc in hexanes) to provide 2- (8-chloro-chloride). N- (2-fluorobenzyl) -1- methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acetate (I-67). 1H NMR (400 MHz, CDCl3): δ 7.47 - 7.43 (m, 2H), 7.34 - 7.29 (m, 1H), 7.23 - 7.19 (m, 1H), 7 , 18 - 7.04 (m, 2H), 6.97 (dd, J = 2.3, 8.7 Hz, 1H), 5.57 - 4.12 (m, 9H), 3.77 - 3 , 74 (m, 3H). Mixture of routines. MS (m / z): 444.1 (M + H) +. Step 2
[00391] 2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acetate (I-67) (1.6 mmol) was dissolved in THF / MeOH / H2O (3: 2: 1, 10 mL) and followed by the addition of LiOH monohydrate (0.408 g, 9.72 mmol). The reaction mixture was stirred at room temperature for 1 hour and diluted with 10 ml of water and acidified to pH = 2.0, Solid was collected and dried to provide 2- (8-chloro-N- (2-fluorobenzyl) acid -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) acetic (I-68). 1H NMR (400 MHz, DMSO): δ 12.73 (s, 1H), 7.71 (t, J = 2.6 Hz, 1H), 7.42 - 7.30 (m, 3H), 7, 24 - 7.15 (m, 2H), 7.07 - 7.02 (m, 1H), 5.41 - 4.10 (m, 9H). Mixture of rotamers. MS (m / z): 430.0 (M + H) +. 3- (2 - ((cyclopentylmethyl) amino) acetamido) -4-methyl fluorobenzoate (I-70)
Caption: water Step 1
[00392] Reagents were added to a 40 mL reaction vessel in the following order: N- (tert-butoxycarbonyl) glycine (1.0 mmol), THF (4.0 mL), N-methyl morpholine (ca 0.2 mL , 2.0 mmol). This was stirred until complete dissolution. Then 2-chloro-4,6-dimethoxy-1,3,5-triazine (1.0 mmol) was added and this solution was stirred for 20 minutes at 50 ° C until a totally white precipitate formed. The precipitate was physically stirred to ensure that all solids were well stirred. Then methyl 3-amino-4-fluorobenzoate (1.3 mmol) was added and the reaction was stirred for 30 minutes at 50 ° C and then at room temperature for 2 hours, and then diluted with 5mL of MeOH and in then purified, and poured into water (rapidly stirred, 25 mL). A white solid was collected and treated with dichloromethane (5 ml) and TFA (5 ml), and heated at 34 ° C for about 1 hour until removal of the Boc group was completed (by LCMS monitoring). The material was concentrated to dryness and subjected to reverse phase chromatography (10 to 40% water / ACN) to provide I-69. The desired fractions were then passed through free base ion exchange polymer binding cartridges to remove any TFA residue (using StratoSpheres product, SPE PL HCO3 MP SPE 0.9 mmol nominal load x 2 units). Mobilizing eluent was MeOH (10 mL of total discharge volume). 1H NMR NMR (400 MHz, d4-MeOH): δ 8.71 (br d, J = 8.0 Hz, 1H), 7.79-7.75 (m, 1H), 7.21 (dd, J = 10.3, 8.2 Hz, 1H), 3.87 (s, 2H), 3.82 (s, 3H). MS (m / z): 227.1 (M + H) +. Step 2
[00393] To a 20 mL reaction vessel, reagents were added in the following order: methyl 3- (2-aminoacetamido) -4-fluorobenzoate (I-69) (0.63 mmol,), AcOH (1 mL), MeOH (4 mL) and cyclopentanecarbaldehyde (1 mmol). After 30 minutes, sodium cyanoborohydride (0.80 mmol) was added. The reaction was stirred 10 minutes, at which time an additional portion of sodium cyanoborohydride was added (0.40 mmol). The reaction was stirred for 2 hours, filtered, and then modified TFA (0.05%) reverse phase chromatography (20 to 60%, water-ACN) was purified and subsequently residual TFA was removed using an attached ion exchange cartridge a polymer (SPE PL HCO3 MP SPE 0.9 mmol nominal charge x 1 unit and 8 mL MeOH as the mobilizing eluant) to provide the amine (I-70) as a glassy solid. 1H NMR NMR (400 MHz, d4-MeOH): δ 8.80 (br d, J = 8.0 Hz, 1H), 7.91-7.86 (m, 1H), 7.32 (dd, J = 11.0, 8.0 Hz, 1H), 4.08 (s, 2H), 3.92 (s, 3H), 3.08 (app d, J = 8.1 Hz, 2H), 2, 31-2.22 (m, 1H), 1.97-1.92 (m, 2H), 1.77-1.64 (m, 4H), 1.36-1.30 (m, 2H). MS (m / z): 309.1 (M + H) +.
[00394] The following intermediates were prepared according to the procedures described for the synthesis of I-70 using the appropriate reagents.
8-chloro-N- (2-fluorobenzyl) -N- (2-hydroxyethyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamide (I-73)
Step 1
[00395] To a 250 mL reaction vessel, reagents were added in the following order: ethanol amine (40.0 mmol), AcOH (8 mL), MeOH (60 mL), and 2-fluorobenzaldehyde (20.0 mmol). After 30 minutes, sodium cyanoborohydride (20.0 mmol) was added in portions over 30 minutes. The reaction was stirred 1 hour, and then partially concentrated (in vacuo) before direct purification by modified acetic acid (0.05%) reverse phase chromatography (5 to 15%, water-ACN) to provide after drying 2- ((2-fluorobenzyl) amino) ethanol (I-72) as a semisolid. 1H NMR (400 MHz, d4-MeOH): δ 7.61 (dt, J = 7.9, 1.8 Hz, 1H), 7.41-7.35 (m, 1H), 7.21 (app t, J = 7.9 Hz, 1H), 7.13 (app t, J = 8.9 Hz, 1H), 4.00 (s, 2H), 3.73 (t, J = 6.2 Hz , 1H), 2.87 (t, J = 6.2 Hz, 1H). MS (m / z): 170.1 (M + H) +. Step 2
[00396] To a 100 mL reaction vessel, reagents were added in the following order: 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxylic acid (I-4 ) (2.0 mmol, THF (10 mL) and N-methyl morpholine (0.72 mL, 5.0 mmol). This was stirred until complete dissolution. Then, 2-chloro-4.6 was added -dimethoxy-1,3,5-triazine (2.2 mmol) and this solution was stirred for 20 minutes at 50 ° C until it formed a totally white precipitate.The precipitate was physically stirred with rapid stirring to ensure that all solids were Then, 2 - ((2-fluorobenzyl) amino) ethanol (I72) (2.4 mmol) was added and the reaction was stirred for 30 minutes at 50 ° C and then diluted with 5 ml of MeOH and then purified by modified acetic acid (0.05%) reverse phase chromatography (30 to 80%, water / ACN) and subsequently recrystallized by MeOH / water 1: 3 (10mL) to provide a white powder (I-73). 1H NMR (400 MHz, DMSO): δ 7.74-7.70 (m, 1H), 7.38-7.28 (m, 5H), 7.04 (dd, J = 8.4 , 1.5 Hz, 1H), 5.40-5.35 (m, 3H), 4.80-4.75 (m, 2H), 4.20 (s, 3H), 3.93 (t, J = 6.2 Hz, 1H) 3.63 (q, J = 6.2 Hz, 1H), 3.55 (q, J = 6.2 Hz, 1H), 3.41 (t, J = 6 , 5 Hz, 1H). Mixture of rotamers. MS (m / z): 416.1 / 418.1 (M + H) + (chlorine isotope standard).
[00397] The following intermediates were prepared according to the procedure described for the synthesis of I-73 using the appropriate reagents.

1,5,5-trimethyl-4,5-dihydro-1H-benzo [g] indazole-3-carboxylic acid (I-78)
Caption: steps
[00398] Intermediate I-78 was prepared according to the procedure described for the preparation of intermediate I-4, starting with 4,4-dimethyl-3,4-dihydronaphthalen-1 (2H) -one. MS (m / z): 257.1 (M + H) +. Example 1
4-Fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid (Compound 1) Step 1
[00399] To a solution of methyl 4-fluoro-3- (2 - ((2-fluorobenzyl) amine) acetamido) benzoate (I-17) (8.2 mmol) and HATU (8.2 mmol) in DMF (30 mL), diisopropylethylamine (30 mmol) and 8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxylic acid (I-5) (7 , 4 mmol) were added to provide a yellowish solution. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was poured onto 300 ml of ice resulting in a precipitate that was collected by vacuum filtration. The solid was then redissolved in EtOAc (600 ml) and washed with 5% Na2CO3, water, brine, dried and concentrated. The solid was purified by trituration in hot MeOH. The resulting solid was filtered and dried. 1H NMR (400 MHz, DMSO): δ 10.14 (m, 1H), 8.79 - 8.46 (m, 1H), 7.80 - 7.66 (m, 1H), 7.56 (m , 1H), 7.50 - 7.29 (m, 3H), 7.22 (m, 3H), 7.06 (m, 1H), 5.38 (m, 3H), 4.82 (d, J = 41.7 Hz, 3H), 4.35 - 4.03 (m, 3H), 3.85 (m, 3H). Mixture of rotamers. Step 2
[00400] The resulting ester (5 mmol) was dissolved in a 4: 1 THF-MeOH (100 mL) and treated with 1N KOH (20 mL). The white suspension was stirred at room temperature for 2 hours resulting in a clear solution. The reaction was stirred at room temperature for an additional 16 hours until the reaction was completed, as indicated by LCMS. The organic solvent was removed in vacuo resulting in a white suspension. The resulting suspension was diluted with water (100 ml) and the pH was adjusted to ~ 5 w / AcOH. The white solid was collected by filtration, washed with water and dried under vacuum for 24 hours. The carboxylic acid product was again purified by stirring in 8: 2 MeOH-water solution (200 ml) at 70 ° C for 2 hours. After cooling to room temperature, the solid was collected by vacuum filtration to provide 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3-c] pyrazol-3-carboxamido) acetamido) benzoyl. 1H NMR (400MHz, DMSO): δ 10.06-10.01 (m, 1H), 8.58-8.56 (m, 1H), 8.51-8.49 (m, 1H), 7, 72-7.69 (m, 1H), 7.58-7.53 (m, 1H), 7.46-7.33 (m, 3H), 7.24-7.12 (m, 3H), 7.08-7.04 (m, 1H), 5.40-5.37 (m, 3H), 4.86-4.76 (m, 3H), 4.24-4.12 (m, 4H ). Mixture of rotamers. MS (m / z): 551.1 (M + H) +.
[00401] In one embodiment, a solution of L-arginine (0.82 mmol) in deionized water (6 mL) was added to 4-fluoro-3- (2- (8-fluoro- N- (2-fluorobenzyl acid) ) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3 carboxamido) acetamido) benzoic (0.82 mmol) in MeOH (50 mL). The clear mixture was stirred for 0.5 hour at room temperature and then concentrated to provide a white semi-solid residue. The residue was taken up in anhydrous acetronitrile (25 ml) and the white suspension was slowly concentrated under reduced pressure. The process was repeated several times (5x) to provide L-arginine salt of 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-di- hydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoyl. Melting point (157 ° C). 1H NMR (400 MHz, DMSO): δ 9.88 - 9.77 (m, 1H), 8.37 - 8.26 (m, 1H), 8.03 - 7.68 (m, 5H), 7 , 66 - 7.51 (m, 2H), 7.48 - 7.31 (m, 3H), 7.15 (m, 6H), 5.43 - 5.35 (m, 2H), 4.86 - 4.72 (m, 3H), 4.22 - 4.10 (m, 4H), 3.26 - 3.18 (m, 1H), 3.17 - 2.99 (m, 2H), 1 , 76 - 1.52 (m, 4H).
[00402] In another embodiment, 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid suspension [4,3-c] pyrazol-3-carboxamido) acetamido) benzoyl (0.84 mmol) in MeOH (20 mL) was heated to 70 ° C. A 1N NaOH solution (0.84 ml) was added dropwise. The clear solution was stirred at room temperature for 1 hour before the solvent was removed under reduced pressure. The crude material was triturated with MeOH and crystallized by suspension in acetonitrile (5 ml) to provide sodium salt of 4-fluoro-3- (2- (8-fluoro-N- (2-fluorobenzyl) - 1-methyl- 1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoyl. Melting point (161 ° C). 1H NMR (400 MHz, DMSO): δ 9.86 - 9.60 (m, 1H), 8.27 - 8.17 (m, 1H), 7.63 - 7.55 (m, 2H), 7 , 48 - 7.31 (m, 2H), 7.26 - 7.04 (m, 5H), 5.42 - 5.37 (m, 2H), 4.84 - 4.74 (m, 3H) , 4.20 - 4.12 (m, 4H). 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid (Compound 2)
[00403] 4-Fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido acid) benzoic acetamido) was prepared following the procedures for Compound 1. Melting point (246 ° C). 1H NMR (400 MHz, DMSO): δ 10.14 - 10.04 (m, 1H), 8.71 - 8.58 (m, 1H), 7.79 - 7.69 (m, 1H), 7 , 62 - 7.51 (m, 1H), 7.47 - 7.36 (m, 2H), 7.29 - 7.02 (m, 5H), 5.42 - 5.32 (m, 2H) , 4.84 - 4.72 (m, 2H), 4.23 - 4.08 (m, 3H), 3.88 - 3.80 (m, 3H). Mixture of rotamers. MS (m / z) (M + H) +, 551.3. Analysis Calculated for C28H21F3N4O5 * 0.15 CH4O: C, 61.09; H, 3.85; N, 10.18. Found: C, 60.97; H, 4.02; N, 10.16.
[00404] Methanol (500 ml) and dichloromethane (500 ml) were added to a mixture of 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4 -dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic (22 mmol) and 2-amino-2-hydroxymethyl-propane-1,3-diol (TRIS; 22 mmol). The reaction was stirred at 25 ° C for 3 hours. The solvent was removed in vacuo, and dichloromethane (500 ml) was added to the solid and concentrated (2x re-applied). The solid was suspended in 100 ml of dichloromethane and collected by vacuum filtration to provide TRIS salt of 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1 acid, 4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoyl. Melting point (125 ° C). 1H NMR (400 MHz, DMSO) δ 10.01 - 9.65 (m, 1H), 8.59 - 8.30 (m, 1H), 7.76 - 6.95 (m, 10H), 5, 53 - 4.00 (m, 9H), 3.58 - 3.10 (m, 10H). Analysis Calculated for C32H32F3N5O8 * 1 H2O: C, 55.73; H, 4.97; N, 10.16. Found: C, 55.74; H, 4.99; N, 10.12.
[00405] In one embodiment, a solution of L-arginine (0.73 mmol) in deionized water (5.8 mL) was added to the suspension of 4- fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic (0.73 mmol) in methanol (70 mL). The mixture was heated to 70 ° C for 0.5 hours. After cooling in, the solvent was removed in vacuo. A 2: 1 mixture of acetonitrile and methanol (20 ml) was added to the solid and concentrated (repeated 2x). The solid was crystallized by suspending the solid residue in a 2: 1 mixture of acetonitrile and methanol (5 mL) and stirring the suspension at room temperature for 24 hours to provide 4- fluoro-3- (2- (8-fluoro) acid -N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic L-arginine salt as a white precipitate. Melting point (206 ° C). 1H NMR (400 MHz, DMSO) δ 9.88 - 9.78 (m, 1H), 8.41 - 8.31 (m, 1H), 8.08 - 7.82 (m, 5H), 7, 68 - 7.52 (m, 2H), 7.47 - 7.36 (m, 1H), 7.28 - 7.04 (m, 6H), 5.43 - 5.30 (m, 3H), 4.81 - 4.70 (m, 3H), 4.14 (s, 4H), 3.26 - 3.21 (m, 2H), 3.16 - 3.01 (m, 2H), 1, 78 - 1.52 (m, 4H).
[00406] In another embodiment, a solution of L-lysine (0.20 mmol) in deionized water (3 mL) was added to a hot suspension of 4-fluoro-3- (2- (8-fluoro-N-) acid (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic (0.20 mmol) in methanol (25mL). The mixture was stirred at room temperature for 16 hours. The solvent was removed in vacuo and the crude material was crystallized by suspending the solid residue in acetonitrile (5 ml). The mixture was stirred at room temperature for an additional 24 hours to provide 4-fluoro-3- (2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4- L-lysine salt dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic as a white precipitate. 1H NMR (400 MHz, DMSO): δ 9.86 - 9.77 (m, 1H), 8.40 - 8.28 (m, 1H), 7.68 - 7.51 (m, 2H), 7 , 48 - 7.36 (m, 1H), 7.29 - 7.02 (m, 7H), 5.43 - 5.36 (m, 2H), 5.36 - 5.31 (m, 1H) , 4.80 - 4.71 (m, 3H), 4.19 - 4.11 (m, 4H), 3.17 - 3.09 (m, 1H), 2.76 - 2.64 (m, 2H), 1.77 - 1.31 (m, 7H); some peaks fall below the peaks of the solvent.
[00407] The following compounds were prepared according to the procedures described in Example 1 using the appropriate intermediates.




Example 2 3- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid (Compound 29 )
Caption: oxalyl chloride Step 1
[00408] To a suspension of 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxylic acid (I-4) (0.20 mmol) in CH2Cl2 (1 mL) a catalytic amount of DMF (2.5 μL) and oxalyl chloride were added dropwise (0.60 mmol) at 0 ° C. The resulting suspension was warmed to room temperature and stirred for 1 hour. The solvent was removed under vacuum until complete dryness (it was necessary to remove oxalyl chloride completely). The obtained residue was dissolved with CH2Cl2 (1 ml) and then dropwise in a solution of methyl 3- (2- (benzylamino) acetamido) -4-fluorobenzoate (I-19) (0.2mmol) in CH2Cl2 ( 1 mL) in the presence of DIEA (0.38 mmol). The reaction mixture was stirred for 15 minutes at room temperature and was then charged with 50 ml of water. The organic layer was separated and the aqueous layer was extracted with CH2Cl2 (50 ml). Combined organic layers were washed with H2O and brine successively, dried over Na2SO4, filtered and concentrated to provide the crude product which was purified by column chromatography (0 to 60% EtOAc in hexanes) to provide 3- (2- (N- benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-methyl fluorobenzoate (29-Int). 1H NMR (400 MHz, CDCl3): δ 9.94 (s, 1H), 9.10 - 8.85 (m, 1H), 7.82 (s, 1H), 7.54 - 7.30 (m , 6H), 7.26 - 7.21 (m, 1H), 7.21 - 7.10 (m, 1H), 7.01 (d, J = 8.3 Hz, 1H), 5.64 - 4.45 (m, 5H), 4.36 - 4.06 (m, 4H), 3.95 (s, 3H). Mixture of rotamers. MS (m / z): 563.1 (M + H) +. Step 2
[00409] methyl 3- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoate ( 29-Int) (0.14 mmol) was dissolved in THF / MeOH / H2O (3: 2: 1, 5.0 mL) and followed by the addition of LiOH monohydrate (0.85 mmol). The reaction mixture was stirred at room temperature for 2 hours. Solvents were removed in vacuo and acidified to pH = 3.0 by adding 3N HCl. Solid was collected and dried to provide 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) acid) acetamido) -5-fluorobenzoic. 1H NMR (400MHz, DMSO): δ 10.11 - 9.96 (m, 1H), 8.67 - 8.53 (m, 1H), 7.78 - 7.66 (m, 2H), 7, 43 - 7.26 (m, 7H), 7.11 - 7.03 (m, 1H), 5.48 - 4.10 (m, 9H). Mixture of rotamers. MS (m / z): 549.1 (M + H) +.
[00410] The following compounds were prepared according to the procedures described in Example 2 using the appropriate intermediate.
Example 2A 3- (2- (9-Chloro-N- (2-fluorobenzyl) -1-methyl-4,5-dihydro-1 H-benzo [2,3] oxepine [4,5-c] acid pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic (Compound 34)
Caption: oxalyl chloride Step 1
[00411] To a suspension of 9-chloro-1-methyl-4,5-dihydro-1H-benzo [2,3] oxepine [4,5-c] pyrazole-3-carboxylic acid (1.8 mmol ) in CH2Cl2 (10 mL) a catalytic amount of DMF (25 µL) and oxalyl chloride (8.9 mmol) was added dropwise at 0 oC. The resulting suspension was warmed to room temperature and stirred for 1 hour. The solvent was removed under vacuum until complete dryness. The obtained residue was dissolved with CH2Cl2 (10 mL) and then dropwise and a solution of methyl 4-fluoro-3- (2 - ((2-fluorobenzyl) amino) acetamido) benzoate (I-17) ( 1.8 mmol) in CH2Cl2 (10 mL) in the presence of DIEA (3.6 mmol). The reaction mixture was stirred for 30 minutes at room temperature and was then charged with 50 ml of water. Organic layer was separated and the aqueous layer was extracted with CH2Cl2 (50 ml). Combined organic layers were washed with H2O and brine successively, dried over Na2SO4, filtered and concentrated to provide the crude product which was purified by column chromatography (0 to 60% EtOAc in hexanes) to provide N- (2- ( (5-acetyl-2-fluorophenyl) amino) -2-oxoethyl) -9-chloro-N- (2-fluorobenzyl) -1- methyl-4,5-dihydro-1H-benzo [2,3] oxepine [4,5-c] pyrazole-3-carboxamide. 1H NMR (400 MHz, CDCl3): δ 10.11 (s, 1H), 8.74 (s, 1H), 7.72 (s, 1H), 7.38 - 7.31 (m, 3H), 7.27 - 7.22 (m, 1H), 7.12 - 6.97 (m, 3H), 5.28 - 4.79 (m, 2H), 4.38 (t, J = 6.4 Hz, 3H), 4.05 - 3.94 (m, 4H), 3.84 (s, 3H), 3.13 (t, J = 6.4 Hz, 2H). MS (m / z): 595.1 (M + H) +. Step 2
[00412] N- (2 - ((5-acetyl-2-fluorophenyl) amino) -2-oxoethyl) -9-chloro-N- (2-fluorobenzyl) -1-methyl-4,5-dihydro- 1H-benzo [2,3] oxepine [4,5-c] pyrazol-3-carboxamide (1.6 mmol) was dissolved in THF / MeOH / H2O (3: 2: 1, 10mL) and followed by addition of monohydrate LiOH (9.7 mmol). The reaction mixture was stirred at room temperature for 4 hours and diluted with 10 ml of water and acidified to pH = 3.0, Solid was collected and dried to produce 3- (2- (9-chloro-N- (2 -fluorobenzyl) -1- methyl-4,5-dihydro-1H-benzo [2,3] oxepine [4,5-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic. 1H NMR (400MHz, DMSO): δ 10.03 (s, 1H), 8.69 - 8.48 (m, 1H), 7.84 - 7.66 (m, 2H), 7.52 - 7, 30 (m, 4H), 7.28 - 7.12 (m, 3H), 5.18 - 4.20 (m, 6H), 4.05 - 3.92 (m, 3H), 3.11 - 2.93 (m, 2H). Mixture of rotamers. MS (m / z): 581.1 (M + H) +. Example 3 3- (2- (8-Chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid (Compound 35)
Caption: KOH (aqueous)
[00413] To a 40 mL reaction vessel, reagents were added in the following order: 2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3- c] pyrazole-3-carboxamido) acetic (I-22) (0.10 mmol), THF (2 mL), and N-methyl morpholine (1.4 mmol). This was stirred until complete dissolution. Then 2-chloro-4,6-dimethoxy-1,3,5-triazine (0.20 mmol) was added and this solution was stirred for 20 minutes at 50 ° C until a completely white precipitate was formed. The precipitate was physically stirred to ensure that all solids were well stirred. Then methyl 3-aminobenzoate (0.30mmol) was added and the reaction was stirred for 30 minutes at 50oC and then diluted with 5 ml of MeOH and then directly purified by modified acetic acid (0.05%) chromatography of reverse phase (30 to 80%, ACN / water). Ester intermediate was used without further purification and was dissolved in 2 ml of MeOH, 2 ml of THF, and treated with 1 ml of 1N aqueous KOH solution (1.0 mmol). After heating the homogeneous solution at 60 ° C for 30 minutes, the reaction was cooled to room temperature, quenched with AcOH (17 N, approximately 0.06 mL) to about pH-6. The reaction was diluted with ethyl acetate (30 ml), rinsed with 2 x 5 ml of water, and the ethyl acetate fraction was partially concentrated to provide 3- (2- (8-chloro-N- (2- fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) acetamido) benzoic as a white solid. 1H NMR NMR (400 MHz, DMSO): δ 11.91 (br s, 1H), 10.15 (br s, 1H), 8.26-8.20 (m, 1H), 7.85 - 7, 62 (m, 3H), 7.48-7.29 (m, 4H), 7.20 (app q, J = 7.5 Hz, 2H), 7.05 (dd, J = 1.7, 8 , 7 Hz, 1H), 5.39-5.35 (m, 2H), 4.84-4.70 (m, 2H), 4.23-4.10 (m, 5H). Mixture of rotamers. MS (m / z): 549.2 / 551.2 (M + H) + (chlorine isotope standard). Example 4 3- (2- (8-chloro-N- (cyclopentylmethyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid ( Compound 36)
Caption: KOH (aqueous)
[00414] To a 40 mL reaction vessel, reagents were added in the following order: 8-chloro-1-methyl -1,4-dihydrochromene [4,3-c] pyrazole-3-carboxylic acid (I-4 ) (0.10 mmol, THF (3 mL) and N-methyl morpholine (1.0 mmol). This was stirred until complete dissolution. Then 2-chloro-4,6-dimethoxy-1,3,5 was added -triazine (0.11 mmol) and this solution was stirred for 20 minutes at 50 ° C to form a completely white precipitate.The precipitate was physically stirred with rapid stirring to ensure that all solids were mixed well. 3- (2 - ((cyclopentylmethyl) amino) acetamido) -4-methyl fluorobenzoate (I-70) (0.10 mmol) was added and the reaction was stirred for 30 minutes at 50 ° C and then diluted with 5 ml MeOH and then purified by modified acetic acid (0.05%) reverse phase chromatography (30 to 80%, water / ACN). The intermediate ester was carried out without further manipulation and was dissolved in 2 mL of MeOH, 2mL of THF, and treated with 1 mL of solution aqueous 1N KOH (1.0 mmol). After heating the homogeneous solution at 60oC for 30 minutes, the reaction was cooled to room temperature, quenched with AcOH (17 N, approximately 0.06 mL) to about pH = 6. The reaction was diluted with 30 mL of ethyl acetate, rinsed with 2 x 5mL of water, and the ethyl acetate fraction was partially concentrated to provide 3- (2- (8-chloro-N- (cyclopentylmethyl) -1-methyl-1,4-dihydrochromene acid [4 , 3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic as a white solid. 1H NMR (400 MHz, DMSO): δ 10.06-10.01 (m, 1H), 8.46-8.40 (m, 1 H), 7.82-7.78 (m, 2 H) , 7.36-7.32 (m, 2H), 7.03-7.00 (m, 1H), 5.32-5.20 (m, 2H), 4.89 (br s, 1H), 4.29 (br s, 2H), 4.24-3.90 (m, 4H), 2.10-2.05 (m, 1H), 1.87-1.67 (m, 6H), 1.091 , 02 (m, 2H). Mixture of rotamers. MS (m / z): 541.2 / 542.2 (M + H) + (chlorine isotope standard). Example 5 3- (2- (8-chloro-N- (cyclopentylmethyl) -1-methyl-1,4-dihydrochromene [4,3-c | pyrazol-3-carboxamido) acetamido) benzoic acid (Compound 37)
Caption: KOH (aqueous)
[00415] 3- (2- (8-chloro-N- (cyclopentylmethyl) -1-methyl -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid was pre- stopped following the procedures described in Example 4, except for the substitution of 3- (2 - ((cyclopentylmethyl) amino) acetamido) methyl benzoate (I-71) in place of 3- (2 - ((cyclopentylmethyl) amino) acetamido) -4-methyl fluorobenzoate (I-70). 1H NMR (400 MHz, DMSO): δ 12.66 (br s, 1H), 10.18-10.14 (m, 1H), 8.23 (app d, J = 10.0 Hz, 1 H) , 7.89-7.12 (m, 5 H), 7.06-7.02 (m, 1H), 5.35-5.25 (m, 2H), 4.89 (br s, 1H) , 4.72 (br s, 2H), 4.43 (br s, 1H), 4.14-4.00 (m, 3H), 1.99-0.96 (m, 9H). Mixture of rodamers. MS (m / z): 523.2 / 525.2 (M + H) + (chlorine isotope standard). Example 6 Acid 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -5- fluorobenzoic (Compound 38)
Step 1
[00416] To a mixture of 2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetic acid (I -22) (0.032 mmol), DIEA (0.16 mmol) and ethyl 3-amino-5-fluorobenzoate (0.05 mmol) in EtOAc (0.5 mL) was added T3P (50% by weight in EtOAc, 0.5 mL) and stirred at room temperature for 30 minutes. HPLC purification gave 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -5 -ethyl fluorobenzoate (38-Int). 1H NMR (400 MHz, CDCl3): δ 9.38 - 8.71 (m, 1H), 7.95 - 7.69 (m, 1H), 7.59 (s, 1H), 7.49 - 7 , 39 (m, 3H), 7.26 - 7.22 (m, 1H), 7.14 (td, J = 1.1, 7.5 Hz, 1H), 7.08 - 7.02 (m , 1H), 7.00 (d, J = 8.7 Hz, 1H), 5.53 (s, 2H), 5.08 - 4.81 (m, 1H), 4.67 - 4.47 ( m, 1H), 4.40 (q, J = 7.1 Hz, 2H), 4.17 (s, 3H), 1.42 (t, J = 7.1 Hz, 3H). MS (m / z): 595.1 (M + H) +. Step 2
[00417] 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -5- ethyl fluorobenzoate (38-Int) (8 mg, 0.0134 mmol) was dissolved in THF / MeOH / H2O (3: 2: 1, 5.0 mL) and followed by the addition of LiOH monohydrate (0.08 mmol ). The reaction mixture was stirred at room temperature for 2 hours. The solvents were removed in vacuo and acidified to pH = 3.0 by adding 3N HCl to provide 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-di acid - hydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -5-fluorobenzoic as a solid. 1H NMR (400MHz, DMSO): δ 10.59 - 10.47 (m, 1H), 7.99 (s, 1H), 7.87 - 7.79 (m, 1H), 7.79 - 7, 72 (m, 1H), 7.53 - 7.43 (m, 1H), 7.43 - 7.36 (m, 3H), 7.32 - 7.23 (m, 2H), 7.14 - 7.08 (m, 1H), 5.51 - 4.13 (m, 9H). Mixture of rotamers. MS (m / z): 567.0 (M + H) +.
[00418] The following compounds were prepared according to the procedures described in Example 6 using the appropriate intermediates.
3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -2-methylpropanoic acid ( Compound 40)
Step 1
[00419] To a mixture of 2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetic acid (I -22) (0.15mmol), DIEA (0.45mmol) and HATU (0.18mmol) in DMF (1ml) ethyl 3-amino-2-methylpropanoate (0.22mmol) was added and stirred at room temperature for 10 minutes. Then, the reaction was diluted with 20 ml of water and extracted with EtOAc (20 ml, twice). Combined organic layers were washed with H2O and brine soaked, dried over Na2SO4, filtered and concentrated to provide the crude product which was purified by column chromatography (0 to 80% EtOAc in hexanes) to provide 3- (2- (8- chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -2-methylpropanoate (40-Int). 1H NMR (400 MHz, CDCl3): δ 7.47 (d, J = 2.4 Hz, 1H), 7.43 - 7.33 (m, 1H), 7.33 - 7.29 (m, 1H ), 7.22 (dd, J = 2.4, 8.7 Hz, 1H), 7.14 (dt, J = 3.8, 7.5 Hz, 1H), 7.08 (t, J = 9.2 Hz, 1H), 6.98 (d, J = 8.7 Hz, 1H), 5.60 - 4.74 (m, 5H), 4.25 - 3.89 (m, 6H), 3.62 - 3.39 (m, 1H), 3.40 - 3.24 (m, 1H), 2.72 - 2.61 (m, 1H), 1.23 - 1.14 (m, 6H ). Mixture of routines. MS (m / z): 543.1 (M + H) +. Step 2
[00420] 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -2- ethyl methylpropanoate (40-Int) (0.125 mmol) was dissolved in THF / MeOH / H2O (3: 2: 1, 5.0 mL) and followed by the addition of LiOH monohydrate (0.75 mmol). The reaction mixture was stirred at room temperature for 2 hours. The solvents were removed in vacuo and then water (5 ml) was added and acidified to pH = 3.0 by adding 3N HCl to provide 3- (2- (8-chloro-N- (2-fluorobenzyl) acid -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -2-methylpropanoic. 1H NMR (400 MHz, DMSO): δ 12.26 (s, 1H), 8.10 - 7.99 (m, 1H), 7.73 - 7.67 (m, 1H), 7.40 - 7 , 29 (m, 3H), 7.24 - 7.16 (m, 2H), 7.09 - 7.02 (m, 1H), 5.43 - 3.89 (m, 9H), 3.32 - 3.06 (m, 2H), 1.06 - 0.97 (m, 3H). Mixture of rotamers. MS (m / z): 515.0 (M + H) +.
[00421] The following compounds were prepared according to the procedures described in Example 7 using the appropriate intermediates.
Example 8 4- (2- (8-Chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy acid -3- fluorobenzoic (Compound 43)
Caption: KOH (aqueous) Step 1
[00422] A 8-chloro-N- (2-fluorobenzyl) -N- (2-hydroxyethyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamide (I- 73) (0.45 mmol) THF (7 mL) and methyl 3-fluoro-4-hydroxybenzoate (0.88 mmol), triphenyl phosphine solid (0.90 mmol) were added, the resulting solution cooled to 00C and finally DIAD (diisopropyl azodicarboxylate) dropwise (0.20mL, AldrichTM commercial stock up to 95% content by weight, 0.96mmol). The internal temperature of the reaction is maintained at 0 oC for 10 minutes, and then allowed to warm to room temperature for 20 minutes. After one hour at room temperature, the reaction was diluted with THF (5 mL), filtered, and directly purified by TFA modified (0.05%) reverse phase chromatography (40 to 90%, water / ACN). All fractions were reduced to dryness under vacuum and subjected to a free base event using polymer immobilized carbonate (SPE-CO3H Varian Cartridge, 0.90 nominal loaded with MeOH mobilizer, 10 mL) to provide the methyl ethyl intermediate as a powder whitish (43-Int). 1H NMR (400 MHz, DMSO): δ 7.78-7.69 (m, 2H), 7.63 (br s, 1H), 7.42-7.38 (m, 3H), 7.18 ( app dt, J = 12.3, 8.0 Hz, 3H), 7.04 (app d, J = 8.0 Hz, 1H), 5.40-5.35 (m, 2H), 5.29 (br s, 1H), 4.83 (br s, 1H), 4.52-4.38 (m, 2H), 4.15-4.10 (m, 2H), 3.93 (s, 3H ) 3.73 (s, 3H). Mixture of rotamers. MS (m / z): 568.2 / 570.2 (M + H) + (chlorine isotope standard). Step 2
[00423] The ester, 4- (2- (8-chloro-N- (2-fluorobenzyl) -1- methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) Methyl fluorobenzoate (43-Int), generated in the previous step was then dissolved in THF (10 ml), MeOH (2 ml), and 1.0 M KOH (2.0 mmol). The resulting homogeneous solution was heated at 60oC for 2 hours. At this point, the reaction was cooled back to room temperature, quenched with 0.12 mL of AcOH (2 mmol, for PH-6 using Whatman-4 colored strip indicator paper to monitor). The reaction was then diluted with water (20 ml) and extracted with ethyl acetate (3 x 100 ml). The organic extracts were again washed with water (2 x 15 mL). The organic extracts were removed to dryness and MeOH / water (10 mL, 9: 1) was allowed to precipitate to provide 4- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1 acid , 4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorobenzoic as a white solid. 1H NMR (400 MHz, DMSO): δ 12.51 (br s, 1H), 7.89-7.19 (m, 9H), 7.07 (dd, J = 1.7, 8.7 Hz, 1H), 5.30-5.15 (m, 3H), 4.85 (br s, 1H), 4.43-4.18 (m, 3H, 4.04-3.99 (m, 3H) , 3.78 (m, 1H) Mixture of rotamers MS (m / z): 554.1 / 556.1 (M + H) + (chlorine isotope standard).
[00424] The following compounds were prepared according to the procedures described in Example 8 using the appropriate intermediates.

Example 9 4- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorobenzoic acid (Compound 55 )
Step 1
[00425] To a solution of 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxylic acid (I-4) (0.16 mmol) in anhydrous DMF So (1 mL) HATU (0.18 mmol), DIEA (0.48 mmol) and methyl 4- (2- (benzylamino) ethoxy) -3-fluorobenzoate (I-52) (0.16 mmol) were added and the resulting mixture was stirred at room temperature until completion (2 hours). The reaction mixture was then diluted with water (10 ml) and extracted with ethyl acetate (20 ml x 3). The organic phase was washed with 1N HCl, water, brine, dried over Na2SO4 and concentrated in vacuo. The crude residue was triturated with MeOH to provide 4- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) - 3-methyl fluorobenzoate as a white solid (55-Int). MS (m / z): 550.1 / 552.1 (M + H) + (chlorine isotope standard). Step 2
[00426] A 4- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3- acid suspension fluorobenzoic (61-int) (20 mg, 0.036 mmol) in a 3: 2: 1 THF - MeOH - H2O solution (2 mL) was treated with a 6M LiOH solution (100 µL) and the resulting mixture was stirred at 35 ° C until complete conversion (3 hours). The reaction mixture was then concentrated under reduced pressure. The crude residue was dissolved in water and 50% acetic acid was added to pH 5 at which time a white solid was separated. The solid was collected by filtration, washed with water and dried under high vacuum to provide 4- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole) acid -3-carboxamido) ethoxy) -3-fluorobenzoic. 1H NMR (400 MHz, DMSO): δ 7.75 - 6.98 (m, 11H), 5.44-5.30 (m, 5H), 4.45 - 3.99 (m, 6H). Mixture of rotamers. MS (m / z): 536.2 / 538.2 (M + H) + (chlorine isotope standard).
[00427] The following compounds were prepared according to the procedures described in Example 9 using the appropriate intermediates.
Example 10 3-Fluoro-4- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c | pyrazol-3-carboxamido) ethoxy acid ) benzoic (Compound 61)
Caption: N, N-diisopropylamine Step 1
[00428] HATU (0.22 mmol) was added to a mixture of 8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxylic acid (I-5) ( 0.20mmol), N, N-diisopropylamine (0.44 mmol) and DMF (2 mL) at room temperature. After stirring for 20 minutes, amine (I-56) (0.20 mmol) in DMF (1 ml) was added and the reaction mixture was stirred for 4 hours at room temperature. The mixture was diluted with H2O (10 ml) and EtOAc (10 ml), the layers were separated, and the H2O layer was washed with EtOAc (x2, 10 ml). The combined organic extracts were washed with H2O (10 ml), brine (10 ml), and then dried (MgSO4). After solvent removal, the crude material was purified by chromatography (solid charge, silica gel, 0 to 60% EtOAc / hexanes) to provide 3-fluoro-4- (2- (8-fluoro-N- (2- fluorobenzyl) - 1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) methyl benzoate (61-Int) as a white solid. 1H NMR (400 MHz, CDCl3): δ 7.84 - 7.64 (m, 2H), 7.43 - 7.29 (m, 1H), 7.21 - 7.13 (m, 1H), 7 , 13 - 7.02 (m, 2H), 7.01 - 6.87 (m, 3H), 5.60 - 5.54 (m, 1H), 5.50 - 5.43 (m, 2H) , 5.30 (s, 3H), 5.01 - 4.94 (m, 1H), 4.45 - 4.37 (m, 2H), 4.37 - 4.30 (m, 1H), 4 , 09 - 4.06 (m, 3H), 3.91 - 3.84 (m, 2H). Step 2
[00429] KOH (1.0 M in H2O, 1.0 mmol) was added to a solution of 3-fluoro-4- (2- (8-fluoro-N- (2-fluorobenzyl) -1-methyl-1 , 4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) methyl benzoate (61-Int) (0.2 mmol) in THF (4.0 mL) and MeOH (1.0 mL ) and the solution was stirred at 50 ° C for 2 hours. After removing solvent (vacuum cleaner), the crude residue was diluted with water (10 mL) and the solution was acidified with acetic acid (to pH ~ 5). The resulting white precipitate was collected by vacuum filtration, washed with H2O (20 mL), and dried overnight in high vacuum to provide 3-fluoro-4- (2- (8-fluoro-N- (2-fluorobenzyl acid) ) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic. 1H NMR (400 MHz, DMSO): δ 7.75 - 7.64 (m, 1H), 7.57 - 7.01 (m, 9H), 5.51 - 5.23 (m, 3H), 4 , 89 - 4.80 (m, 1H), 4.49 - 4.29 (m, 3H), 4.13 - 3.98 (m, 3H), 3.83 - 3.72 (m, 1H) . MS (m / z): 538.2 (M + H) +. Mixture of rotamers.
[00430] The following compound was prepared according to the procedures described in Example 10 using the appropriate intermediates.
Example 11 3 - ((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino acid) benzoic (Compound 63)
Legend: Periodinano Dess-Martin; 3-methyl amino benzoate; water Step 1
[00431] To a 40 mL reaction vessel were charged 8-chloro-N- (2-fluorobenzyl) -N- (2-hydroxyethyl) -1-methyl-1,4-dihydrochromene [4,3-c ] pyrazole-3-carboxamide (I-73) (0.10 mmol) and CH2Cl2 (10 mL) followed by the addition of commercial Periodinane Dess-Martin (Sigma-Aldrich, 1,1,1-Tris (acetyloxy) - 1, 1-dihydro-1,2-benziodoxol-3- (1H) -one, 0.20mmol) and buffered with solid sodium bicarbonate (0.20mmol). The milky white suspension was rapidly stirred for 2 hours and then the reaction was then directly diluted with ethyl acetate (150 ml) and washed with water (3 x 15 ml). The resulting organic extract was concentrated to a residue (in a vacuum) and used directly without delay or other manipulation. The residue was dissolved in 9: 1 MeOH / AcOH (5 ml) and treated with methyl-3-amino benzoate (0.40 mmol). The resulting reaction was then stirred at room temperature for 30 minutes and then treated with sodium cyanoborohydride (1.0 mmol, in portions over 30 minutes) followed by stirring at room temperature for an additional hour. The resulting reaction was diluted with ethyl acetate (150 ml) and washed with water (3 x 25 ml). The resulting organic extracts were concentrated in vacuo and then directly subjected to reverse phase chromatography using modified TFA (0.05%) water / ACN (35 to 80%). All fractions were reduced to dryness under vacuum and subjected to a free base event using polymer immobilized carbonate cartridge (SPE-CO3H Varian, 0.90 nominal loaded with MeOH mobilizer, 10 mL) to provide 3- ((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoate (63 -Int) as a white solid. 1H NMR (400 MHz, d4-MeOH): δ 7.48-7.19 (m, 8H), 7.03-6.92 (3H), 7.18 (app d, J = 8.0 Hz, 1H), 6.62 (app dt, J = 12.2, 8.0 Hz, 1H), 5.30-5.20 (m, 2H), 4.83 (br s, 2H), 4.21 (t, J = 5.0 Hz, 2H), 3.85 (s, 3H), 3.74 (s, 3H), 3.38 (t, J = 5.0 Hz, 2H). Mixture of rotamers. MS (m / z): 549.2 / 551.2 (M + H) + (chlorine isotope standard). Step 2
[00432] methyl 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoate (63-Int) (0.075 mmol) was dissolved in THF (3 mL), MeOH (2 mL), and 1.0 M KOH (1.0 mmol). The resulting homogeneous solution was heated at 60oC for 2 hours. At this point, the reaction was cooled to room temperature, quenched with AcOH (1.2 mmol, to pH = 6 using Whatman-4 colored strip indicator paper to monitor). The reaction was then carried out with water (5 ml) and ethyl acetate (3 x 20 ml) was extracted. The organic extracts were again washed with water (2 x 5 mL). The organic extracts were removed to dryness and allowed to precipitate from MeOH / water (3 mL, 9: 1) to provide 3 - ((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1 acid, 4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic as a white solid. 1H NMR (400 MHz, d4-MeOH): δ 7.48-7.10 (m, 7 H), 7.02-6.90 (m, 3H), 6.61 (br d, J = 8, 0 Hz, 1H), 5.40-5.20 (m, 2H), 4.80 (br s, 2H), 4.21 (t, J = 7.5 Hz, 2H), 3.91 (br s, 3H), 3.38 (t, J = 7.5 Hz, 2H). Mixture of rotamers. MS (m / z): 535.2 / 537.2 (M + H) + (chlorine isotope standard). Example 12 Acid 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4- fluorobenzoic (Compound 64)

[00433] 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorobenzoic acid was prepared according to the procedures described in Example 3 for 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole acid -3-carboxamido) acetamido) benzoyl, using methyl 3-amino-4-fluorobenzoate in place of methyl-3-amino benzoate in step 1. MS (m / z): 567.2 / 569.1 (M + H ) + (chlorine isotope standard); rt = 1.734. Example 13 3 - ((2- (8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino acid) -4- fluorobenzoic (Compound 65)

[00434] HATU (1.2 mmol) was added to a mixture of 8-chloro-1-methyl -1,4-dihydrochromene [4,3,3 c] pyrazole -3-carboxylic acid (I-4) ( 1.1 mmol), Hunig's base (2.4 mmol) and DMF (5 mL). A solution of methyl 4-fluoro-3 - ((2 - ((3-fluorobenzyl) amino) ethyl) amino) benzoate (I-46) (1.1 mmol) in DMF (3 mL) was added and the mixture stirred at room temperature for 4 hours. The mixture was diluted with water and ethyl acetate. The layers were separated and the aqueous phase was washed with ethyl acetate. The combined organic extracts were washed with water, brine, and then dried over MgSO4. The material was purified by chromatography (silica, 0 to 60% ethyl acetate / hexanes) to provide 3 - ((2- (8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-di -hydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-methyl fluorobenzoate (65-Int). 1H NMR (400 MHz, DMSO): δ 7.61 - 7.29 (m, 2H), 7.23 - 6.86 (m, 8H), 6.20 - 5.89 (m, 1H), 5 , 45 - 4.65 (m, 4H), 4.14 - 3.95 (m, 4H), 3.84 - 3.77 (m, 3H), 3.58 - 3.46 (m, 1H) , 3.46 - 3.37 (m, 2H). Step 2
[00435] 1N KOH (4 mmol, 5 equiv.) Was added to a solution of 3 - ((2- (8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-methyl fluorobenzoate (65-Int) (0.80 mmol) in THF (4 mL) and MeOH (2 mL) and the solution was stirred at 50 ° C for 2 hours. The solvent was removed and the crude residue was diluted with water. The aqueous solution was acidified with acetic acid (pH ~ 5) to provide the title compound as a white precipitate. 1H NMR (400 MHz, DMSO): δ 7.61 - 6.76 (m, 10H), 6.21 - 5.77 (m, 1H), 5.46 - 3.94 (m, 9H), 3 , 56 - 3.38 (m, 2H). Mixture of rotamers. MS (m / z): 537.2 (M + H) +.
[00436] The following compounds were prepared according to the procedures described in Example 13 using the appropriate intermediates.

Example 14 4 - ((N-benzyl -8-chloro -1- methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid (Compound 75)
Step 1
[00437] HATU (43.1 mmol) (Oakwood) was added to a solution of 8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxylic acid (I- 4) (39 mmol) and DIEA (15 mL) (Aldrich) in DMF (300 mL) at room temperature. After stirring for 20 minutes, methyl 4- ((benzylamino) methyl) benzoate (I-23) (39 mmol) was added neat as an oil followed by a rinse of DMF (20 mL) and the reaction mixture was stirred for 2 hours. The mixture was poured onto ice and the resulting precipitate was filtered, rinsed with H2O (~ 200 ml). The solid was dissolved in DCM (~ 300 ml), washed with NaHCO3 (saturated), H2O and dried (MgSO4). The raw material was purified by chromatography (silica gel, charged pure with DCM rhinse, 10-100% EtOAc / Hexanes) and after removal of the solvent, an oil was obtained. The oil was suspended in EtOH (~ 100 ml) and the mixture was heated to reflux while stirring. After stirring for 1 hour, the mixture was cooled to room temperature and the resulting solid was filtered and rinsed with cold EtOH (~ 50 ml). The filtrate was concentrated (aspirator) and crystallization was repeated to provide 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) methyl benzoate. 1H NMR (400 MHz, DMSO): δ 7.99 - 7.87 (m, 2H), 7.75 - 7.65 (m, 1H), 7.45 - 7.22 (m, 8H), 7 , 12 - 7.02 (m, 1H), 5.48 - 5.41 (m, 2H), 5.30 - 5.21 (m, 2H), 4.64 - 4.55 (m, 2H) , 4.19 - 4.05 (m, 3H), 3.85 (s, 3H). MS (m / z): 502.0 / 504.0 (M + H) + (chlorine isotope standard). Step 2
[00438] 1.0M potassium hydroxide (53 mmol) was added to a solution of 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido) methyl) methyl benzoate (17.5 mmol) in THF (56 ml) and methanol (14 ml) and the mixture was heated at 50 ° C for 2 hours. After cooling to room temperature, the solvent was removed (vacuum). The crude residue was diluted with water and the aqueous solution was acidified with acetic acid (to pH ~ 6) resulting in a precipitate. The precipitate was collected by vacuum filtration and dried overnight under high vacuum. The solid was collected and EtOH (125 ml) was added. The mixture was stirred at 85 ° C for 2 hours and then cooled to room temperature. The solid was filtered, rinsed with cold EtOH (~ 75 mL) and dried overnight under high vacuum to provide 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4 , 3-c] pyrazol-3-carboxamido) methyl) benzoyl. 1H NMR (400 MHz, DMSO): δ 7.96 - 7.85 (m, 2H), 7.74 - 7.67 (m, 1H), 7.42 - 7.20 (m, 8H), 7 , 12 - 6.97 (m, 1H), 5.46 - 5.42 (m, 2H), 5.28 - 5.21 (m, 2H), 4.62 - 4.49 (m, 2H) , 4.18 - 4.07 (m, 3H). Mixture of rotamers. MS (m / z) 488.2 / 490.2 (M + H) + (chlorine isotope standard).
[00439] Methanol (400 mL) was added to a mixture of 4- ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido acid ) methyl) benzoic (8.20 mmol) and TRIS (8.20 mmol). The reaction was heated to 70 ° C for 0.5 hour. After cooling to room temperature, the solvent was removed in vacuo. The residue was sonicated in dichloromethane (10 ml) and concentrated again. The resulting white solid was dried under vacuum pump overnight. The crude material was crystallized by suspending the solid residue in a 4: 1 mixture of acetonitrile and methanol (5 ml). The mixture was stirred at room temperature for 24 hours to provide 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido acid salt) ) methyl) benzoic TRIS as a white precipitate. Melting point (195.6 oC). 1H NMR (400 MHz, DMSO): δ 7.92 - 7.80 (m, 2H), 7.78 - 7.64 (m, 1H), 7.41 - 7.19 (m, 8H), 7 , 13 - 7.00 (m, 1H), 5.44 (s, 2H), 5.25 - 5.14 (m, 2H), 4.61 - 4.48 (m, 2H), 4.18 - 4.03 (m, 3H), 3.39 (s, 7H).
[00440] In one embodiment, 4 - (((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid (30, 6 mmol) was added to 150 ml of acetone to provide a white suspension. To the suspension, 75 ml of melamine (34 mmol) water solution was gradually added. The resulting mixture was stirred at 50 ° C for 4 hours and then at room temperature for 12 hours to provide 4 - ((N-benzyl-8-chloro-1-methyl -1,4-dihydrochromene [4,3- c] pyrazol-3-carboxamido) methyl) benzoic meglumine monohydrate salt as a white solid, which was dehydrated at about 71 ° C as determined by DSC.
[00441] In another mode, another heating of the meglumine monohydrate salt to 80oC, 0% relative humidity in an oven for 30 minutes provided a white crystalline solid having a melting point (start Tm) of 167.5oC . Alternatively, 4 - ((N-benzyl-8-chloro-1-methyl -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid (0 , 1 mmol) was added to 1 mL 1: 1 (v / v) acetone / water. The resulting mixture was equilibrated at 50 ° C for one week to provide a white solid, which was dehydrated at about 61 ° C as determined by DSC.
[00442] In yet another embodiment, methanol (100 mL) was added to 4 - (((N-benzyl-8-chloro-1-methyl -1,4-dihydrochromene [4,3-c] pyrazole-3 acid) -carboxamido) methyl) benzoyl (4.1 mmol) and meglumine base (4.1 mmol). The resulting suspension was refluxed at 80oC for 24 hours and then cooled to room temperature to provide 4 - ((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene mglumin salt) [4, 3-c] pyrazol-3-carboxamido) methyl) benzoyl as a white solid. Melting point (180.6 oC).
[00443] The following compounds were prepared according to the procedures described in Example 14 using the appropriate intermediates.




Example 15 4- Fluoro-3- (((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole -3-carboxamido) ethyl acid) amino) benzoic (Compound 100)
Legend: N, N-diisopropylethylamine; water Step 1
[00444] HATU (1.2 mmol) was added to a mixture of 8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxylic acid (I-5) ( 1.1 mmol), Hunig's Base (2.4 mmol) and DMF (5 mL). A solution of methyl 4-fluoro-3 - ((2 - ((3-fluorobenzyl) amino) ethyl) amino) benzoate (I46) (1.1 mmol) in DMF (3 mL) was added and the reaction mixture stirred at room temperature for 4 hours. The mixture was diluted with water and ethyl acetate. The layers were separated and the aqueous phase was washed with ethyl acetate. The combined organic extracts were washed with water, brine, and then dried over MgSO4. The material was purified by chromatography (silica, 0 to 60% ethyl acetate / hexanes) to provide 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1 , 4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) methyl benzoate (100-Int). Step 2
[00445] 1N KOH (4 mmol, 5 equiv.) Was added to a solution of 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4 -dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) methyl benzoate (100-Int) (0.80 mmol) in THF (4 mL) and MeOH (2 mL) and the solution it was stirred at 50 ° C for 2 hours. The solvent was removed and the crude residue was diluted with water. The aqueous solution was acidified with acetic acid (pH ~ 5) to provide 4-fluoro-3 - (((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene acid [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic as a white precipitate. 1H NMR (400 MHz, DMSO): δ 7.61 - 7.26 (m, 2H), 7.20 - 6.82 (m, 9H), 6.11 - 5.81 (m, 1H), 5 , 43 - 4.70 (m, 4H), 4.15 - 3.94 (m, 4H), 3.53 (t, J = 6.6 Hz, 1H), 3.44 - 3.39 (m , 2H). MS (m / z): 537.2 (M + H) +; r.t. = 1.618; Elemental Analysis: calculated to 0.50, C28H22F3N4O * 0.52 H2O: C, 61.65; H, 4.43; N, 10.45); Found: C: 61.75; H: 4.21; N: 10.31.
[00446] 4-Fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido acid ) ethyl) amino) benzoic (7.5 mmol) added to a solution of TRIS (7.5 mmol) in MeOH (400 mL). The reaction was stirred at 60 ° C for 30 minutes. After cooling to room temperature, the solvent was removed and the raw material was azeotroped with dichloromethane (2x). The resulting clear solid was suspended in ethyl acetate (200 ml). The suspension was stirred at room temperature for 24 hours. The resulting solid was collected by filtration to provide 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene salt [4,3 -c] pyrazol-3-carboxamido) ethyl) amino) benzoyl TRIS as a solid. Melting point (160 ° C). 1H NMR (600 MHz, DMSO): δ 7.62 - 6.70 (m, 10H), 5-90-5.80 (m, 1H), 5.45 - 5.15 (m, 2H), 4 , 75-4.55 (m, 2H), 4.16 - 3.5 (m, 4H), 3.41-3.39 (m, 1H), 3.30 - 3.25 (m, 13H) . Anal.Calcd for C32H34F3N5O7 * 1.3 H2O: C, 56.36; H, 5.42; N, 10.27 * 1 H2O Found: C, 56.25; H, 5.29; N, 10.32.
[00447] A solution of L-arginine (0.20 mmol) in deionized water (3 mL) was added to a 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) acid suspension -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic (0.20 mmol) in MeOH (12 mL). The mixture was stirred at 70 ° C for 0.5 hour and then stirred for 2 hours at room temperature. The solvent was removed in vacuo and the crude material was crystallized by suspending the solid residue in acetonitrile (5 ml). The mixture was stirred at room temperature for an additional 24 hours to provide L-arginine salt of 4-fluoro-3 - ((2- (8-fluoro-N- (3-fluorobenzyl) - 1-methyl-1,4 -dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic as a white precipitate. Melting point (161 ° C). Example 16 Acid 3 - ((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole -3-carboxamido) ethyl) amino) -4- fluorobenzoic (Compound 101)
Legend: NN-diisopropylethylamine; Water.
[00448] Acid 3- ((2- (8-chloro -N- (2-fluorobenzyl) -1- methyl -1,4-dihydrochromene [4,3-c] pyrazole -3-carboxamido) ethyl) amino ) -4-fluorobenzoic was prepared according to the procedure described in Example 13 using the appropriate intermediates. MS (m / z): 553.2 / 555.2 (M + H) + (chlorine isotope standard); r.t. = 1.871. BIOLOGICAL TESTS Human GST-FXR LBD coactivator Interaction Assay
[00449] The FXR HTRF assay is a biochemical assay measuring the interaction between FXR and a co-activating protein (SRC1). The linker-induced interaction with a co-activating protein is a critical step in transcriptional activation by FXR. Thus, this is an assay designed to measure the FXR agonist activity of the test compounds.
[00450] Human Receptor ligand binding domain Combinant Farnesoid X (FXR) (amino acids 193-472) fused to the protein purified by glutathione S-transferase (GST) (GST-FXR LBD) was acquired (Invitrogen). The linker-dependent interaction between GST-FXR LBD and a peptide derived from Steroid Receptor Coactivator-1 (SRC-1) was monitored by Fluorescence Resonance Energy Transfer (FRET). GST-FXR LBD was mixed with a biotin-labeled SRC-1 peptide (Sequence: Biotin-CPSSHSSLTERHKILHRLLQEG -SPS-CONH2, American Peptide) in assay buffer (50 mM Tris ^ HCl, pH 7.4, 50 mM NaCl, 1 mM TCEP and 0.2% bovine serum albumin) and seeded on 384 black Proxi plates (Greiner Bio-One). Test compounds (in DMSO solution) and detection reagents (anti-GST-Cryptate labeled antibody and Streptavidin-XL665 conjugate; CisBio) were added in assay buffer containing 50 mM KF. The plates are incubated at room temperature in the dark for 2.5 hours, before reading in an Envision (PerkinElmer) at 665 nm and 590 nm. The results of the HTRF test were calculated from the ratio of 665 nm / 590 nm (ratio = (A665nm / A590nm) x 104) and expressed in Delta F% = (Sample ratio - Negative ratio) / Negative ratio x 100.
[00451] A negative control (without Streptavidin-XL665) was conducted with each assay and the base fluorescence was represented. A reference agent of FXR, (E) -3- (2-chloro-4 - (((3- (2,6-dichlorophenyl) - 5-isopropylisoxazol-4-yl) methoxy) styryl) benzoic acid (Compound GW4064), was included in each experiment as a positive control. The effectiveness of each test compound was compared to that of GW4064. In each concentration, the relative activity of the test compound was expressed as% Response = (Ramostra - RDMSO) / (Rpositiva - RDMSO), where Ramostra is the HTRF response (expressed in Delta F%) for the compound test, Rpositiva is the maximum response for GW4064 in saturation concentrations, and RDMSO is the response for DMSO control. EC50 values were calculated using GraphPad Prism (GraphPad Software) using non-linear regression curve fitting (log (agonist) vs. response - variable slope (four parameters)).
[00452] Table 1 summarizes the EC50 values for the compounds of the invention in human GST-FXR LBD coactivator interaction assay. Table 1

na = not determined
[00453] It is understood that the examples and modalities described here are for illustrative purposes only and that several modifications or changes in the light of them will be suggested for those skilled in the art and should be included in the spirit and competence of this request and the scope of the attached claims . All publications, patents, and patent applications cited here are hereby incorporated by reference for all purposes.

权利要求:
Claims (21)
[0001]
1. Compound, characterized by the fact that it presents Formula (I),
[0002]
2. Compound, according to claim 1, characterized by the fact that it presents Formula (II):
[0003]
3. Compound according to claim 1 or 2, characterized by the fact that R0 is selected from: * 3-CH2C (CH3) 2-, * 3-CH2CH (CH3) -, and * 3-cyclopropane-1, 1, - di-il, where "* 3" indicates the connection point from R0 to L2.
[0004]
4. Compound, according to claim 1 or 2, characterized by the fact that it presents Formula (III):
[0005]
Compound according to any one of claims 1 to 4, characterized by the fact that L2 is selected from - (CH2) -, * 2-CH2C (O) NH-, * 2- (CH2) 2O-, and * 2- (CH2) 2NH-; where "* 2" indicates the connection point from L2 to N.
[0006]
A compound according to any one of claims 1 to 5, characterized in that each R2 is independently fluorine or methyl.
[0007]
7. A compound according to any one of claims 1 to 6, characterized by the fact that each R3 is independently selected from hydrogen, fluorine, chlorine, and methyl.
[0008]
A compound according to any one of claims 1 to 7, characterized by the fact that: Z1 is selected from CH, CF, CCH3, and N; Z2 is selected from CH, CF, CCl, and CCH3; Z3 is selected from CH, CF, CCl, and CCH3; and Z4 is CH or N.
[0009]
9. Compound, according to claim 1, characterized by the fact that it presents Formula (IV):
[0010]
10. Compound according to claim 1, characterized by the fact that it is selected from: 4-fluorine-3- (2- (8-fluorine-N- (2-fluorobenzyl) -1-methyl-1 acid , 4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic; 4-fluoro-3- (2- (8-fluoro-N- (3-fluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid ; 3- (2- (8-chloro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluoro-benzoic acid; 4-fluoro-3- (2- (1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (N- (2-fluorobenzyl) -1,8-dimethyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (N- (2-fluorobenzyl) -1,6-dimethyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-fluorine-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (7-fluorine-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (7-Chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (6,8-difluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (N- (2-fluorbenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (2-fluorbenzyl) -1,7-dimethyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1,6-dimethyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-6-fluorine-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid ; 4-fluoro-3- (2- (N- (2-fluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; Acid 3- (2- (7,8-difluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4- fluorobenzoic; Acid 3- (2- (7,8-difluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4- fluorobenzoic; 3- (2- (N-benzyl-7,8-difluor-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorbenzoic acid; 4-fluoro-3- (2- (8-fluoro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid ; 4-Fluoro-3- (2- (N- (2-fluoro-benzyl) -1-methyl-4,5-dihydro-1H-pyrazolo [4,3-h] quinoline-3-carboxamido) acetamido) benzoic acid ; 4-Fluoro-3- (2- (N- (2-fluoro-benzyl) -1-methyl-4,5-dihydro-1-pyrazolo [3,4-f] quinoline-3-carboxamido) acetamido) benzoic acid ; (S) -4-Fluorine-3- (2- (8-fluorine-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazole-3-carboxamido acid ) propanamido) benzoic; 3- (2- (8-chloro-N- (2-fluorbenzyl) -1-methyl-4,5-dihydro-1H-benzo [g] indazole-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-7-fluorine-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid ; 4-Fluoro-3- (2- (N- (3-fluoro-benzyl) -1,5,5-trimethyl-4,5-dihydro-1H-benzo [g] indazole-3-carboxamido) acetamido) benzoic acid ; 4-fluoro-3- (2- (N- (3-fluorbenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (3-fluorbenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluoro-benzoic acid; 3- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorbenzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluoro-benzoic acid; 3- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluoro-benzoic acid; 3- (2- (N-benzyl-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorbenzoic acid; 3- (2- (9-Chloro-N- (2-fluorobenzyl) -1-methyl-4,5-dihydro-1H-benzo [2,3] oxepine [4,5-c] pyrazole-3 acid -carboxamido) acetamido) -4-fluorobenzoic; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (cyclopentylmethyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluorbenzoic acid; 3- (2- (8-chloro-N- (cyclopentylmethyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) benzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -5-fluorbenzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-methylbenzoic acid; 3- (2- (8-chloro-N- (2-fluorbenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -2-methylpropanoic acid; Acid 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -2,2- dimethylpropanoic; Carboxylic acid 1 - ((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) methyl) cyclopropane ; 4- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorbenzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic acid; 4- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic acid; N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-methylbenzoic acid; 4- (2- (8-Chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5- acid dimethylbenzoic; 3- (2- (8-chloro-N- (2-fluorbenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -4-fluoro-benzoic acid; 4- (2- (8-chloro-N- (cyclopentylmethyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5-dimethylbenzoic acid; 4- (2- (8-Chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5- acid diflúorbenzoic; 4- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3- (trifluoromethyl) acid ) benzoic; 4- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5-difluoro-benzoic acid; 3,5-Diflúor-4- (2- (8-fluor-N- (2-fluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy acid ) benzoic; 4- (2- (N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3,5-difluoro-benzoic acid; 4- (2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorbenzoic acid; 4- (2- (N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorbenzoic acid; 4- (2- (N-benzyl-7,8-difluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorbenzoic acid; 3-fluoro-4- (2- (8-fluoro-N- (3-fluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic acid ; 4- (2- (7,8-difluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3- fluorobenzoic; 4- (2- (8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3-fluorbenzoic acid; 3-fluoro-4- (2- (8-fluoro-N- (2-fluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) benzoic acid ; 4- (2- (7,8-difluoro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethoxy) -3- fluorobenzoic; 3 - ((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic acid; 3- (2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) acetamido) -4-fluoro-benzoic acid; 3 - (((2- (8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino acid -4 -fluorbenzoic; 3 - (((2- (8-chloro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino acid -4 -fluorbenzoic; 4-Fluoro-3 - (((2- (8-fluoro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl acid) amino) benzoic; Acid 3 - ((2- (7,8-difluoro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-fluoro-benzoic; 4-Fluoro-3 - (((2- (8-fluoro-N- (2-fluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl acid) amino) benzoic; 4-fluoro-3 - ((2- (N- (2-fluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic acid; 3 - ((2- (N-benzyl-8-fluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-fluoro-benzoic acid; 3 - (((2- (N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-fluorbenzoic acid; Acid 3 - ((2- (7,8-difluoro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-fluoro-benzoic; 3 - (((2- (N-benzyl-7,8-difluoro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) -4-fluoro-benzoic acid ; 4 - (((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - (((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorbenzoic acid; 4 - (((8-fluoro-N- (3-fluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-Chloro-N- (3,5-difluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - (((N-benzyl-8-fluor-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorbenzoic acid; 4 - ((8-chloro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-Chloro-N- (2,3-difluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-Chloro-N- (3-fluor-5-methylbenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - (((N- (3,5-difluoro-benzyl) -8-fluor-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - (((8-chloro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorbenzoic acid; 3-fluoro-4 - (((8-fluoro-1-methyl-N- (3-methylbenzyl) -1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - (((N - ((1H-indol-5-yl) methyl) -8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acid) benzoic; 5 - ((8-chloro-N- (3-fluorbenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) picolinic acid; 4 - (((8-Chloro-N - (((5-fluoro-pyridin-3-yl) methyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acid) benzoic; 4 - (((8-Chloro-N - (((5-chloropyridin-3-yl) methyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl acid) benzoic; 4 - (((8-Chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -2-fluorbenzoic acid; 4 - (((N-benzyl-8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -2-fluorbenzoic acid; N-benzyl-N- (4-carbamoylbenzyl) -8-chloro-1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamide; 4 - ((8-Chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - (((8-Chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorbenzoic acid; 3-fluoro-4 - (((8-fluoro-N- (2-fluoro-benzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - ((8-Chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) benzoic acid; 4 - (((8-Chloro-N - (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -3-fluorobenzoic acid 4- ((8-chloro-N- (2-fluorbenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -2-fluorbenzoic; 6 - ((8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) nicotinic acid; 5 - (((8-chloro-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) methyl) -6-methylpicolinic acid; 4-Fluorine-3 - (((2- (8-fluorine-N- (3-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) benzoic; and Acid 3 - (((2- (8-chloro-N- (2-fluorobenzyl) -1-methyl-1,4-dihydrochromene [4,3-c] pyrazol-3-carboxamido) ethyl) amino) - 4-fluorbenzoic; or a pharmaceutically acceptable salt thereof.
[0011]
A compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized by the fact that it is 4 - ((N-benzyl-8-chloro-1-methyl-1,4-di-idrochromene) [4 , 3-c] pyrazol-3-carboxamido) methyl) benzoic.
[0012]
12. A compound according to claim 11, characterized by the fact that it is TRIS salt of 4 - ((N-benzyl-8-chloro-1-methyl-1,4-di-idrochromene) [4, 3-c] pyrazol-3-carboxamido) methyl) benzoic.
[0013]
13. A compound according to claim 11, characterized by the fact that it is 4- ((N-benzyl-8-chloro-1-methyl-1,4-di-idrochromene) meglumine monohydrate salt [ 4,3-c] pyrazol-3-carboxamido) methyl) benzoic.
[0014]
14. A compound according to claim 11, characterized by the fact that it is a 4 - ((N-benzyl-8-chloro-1-methyl-1,4-di-idrochromene) meglumine salt [4, 3-c] pyrazol-3-carboxamido) methyl) benzoyl.
[0015]
15. Compound, characterized by the fact that it presents Formula (V)
[0016]
16. Pharmaceutical composition, characterized in that it comprises a therapeutically effective amount of a compound, as defined in any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
[0017]
17. Combination, characterized in that it comprises a therapeutically effective amount of a compound, as defined in any one of claims 1 to 14, and a second therapeutic agent.
[0018]
18. Use of a therapeutically effective amount of a compound, as defined in any one of claims 1 to 14, or of a pharmaceutically acceptable salt thereof, characterized by the fact that it is for the manufacture of a medicament for the treatment of a condition mediated by farnesoid X (FXR) receptors, and said FXR-mediated condition is a liver disease selected from intrahepatic cholestasis, estrogen-induced cholestasis, drug-induced cholestasis, pregnancy cholestasis, cholestasis associated with parenteral nutrition, cholestasis progressive family disease (PFIC), Alagille syndrome, primary biliary cirrhosis (PBC), primary sclerosing cholangitis, ductopenic liver transplant rejection, graft versus host disease associated with liver transplantation, cystic fibrosis kidney disease, non-adipose liver disease alcoholic (NAFLD), non-alcoholic steatoepatitis (NASH), alcoholic liver disease, and liver disease associated with parenteral nutrition; or a gastrointestinal disease selected from bile acid malabsorption, bile reflux gastritis and inflammatory bowel disease.
[0019]
19. Use of a therapeutically effective amount of a compound, as defined in any one of claims 1 to 14, or of a pharmaceutically acceptable salt thereof, characterized by the fact that it is for the manufacture of a medicament to treat an FXR-mediated condition , and the FXR-mediated condition is a gastrointestinal disease selected from bile acid malabsorption, bile reflux gastritis, and inflammatory bowel disease.
[0020]
20. Use, according to claim 18, characterized by the fact that said condition mediated by FXR is non-alcoholic adipose liver disease (NAFLD) or non-alcoholic steatoepatitis (NASH).
[0021]
21. Use of a therapeutically effective amount of a compound, as defined in any one of claims 1 to 14, or of a pharmaceutically acceptable salt thereof, characterized by the fact that it is for the manufacture of a medicament for diabetic nephropathy.

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

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

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2019-10-01| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

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2020-12-01| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2021-02-09| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 04/11/2014, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

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US201361900013P| true| 2013-11-05|2013-11-05|

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US61/900,013|2013-11-05|

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