Amide compounds

Abstract

Provided are compounds of formula (I) or stereoisomers or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising same, and the use thereof as androgen receptor (AR) antagonists in the treatment or prevention of androgen receptor-mediated diseases, preferably for the treatment of androgen receptor-mediated alopecia, such as androgenetic alopecia.

Description

Amide compounds

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This disclosure claims priority to and the benefit of the following Chinese patent applications filed with the State Intellectual Property Office of China: Chinese Patent Application No. 202311282820.X filed on September 28, 2023, and Chinese Patent Application No. 202410900706.7 filed on July 5, 2024. The entire text of the above patent applications is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure relates to the field of medicinal chemistry, and in particular to a class of amide compounds or stereoisomers or pharmaceutically acceptable salts thereof, pharmaceutical compositions containing the same, and their use as androgen receptor (AR) antagonists. Background Art

[0004] The androgen receptor (AR) is a member of the nuclear receptor family. It comprises four major regions: the N-terminal active transcriptional control domain (NTD), the DNA binding domain (DBD), the hinge region, and the C-terminal ligand binding domain (LBD). Its activation is closely associated with the development of diseases such as benign prostatic hyperplasia, prostate cancer, seborrheic dermatitis, acne, premenstrual syndrome, lung cancer, polycyclic ovarian syndrome, hirsutism, and alopecia. Therefore, the AR is an important target in multiple areas of drug discovery.

[0005] Androgenic alopecia (AGA) is currently the most common type of hair loss clinically diagnosed, but its specific pathogenesis remains incompletely understood. Most researchers believe it is related to androgen metabolism. Studies have shown that elevated expression of the androgen receptor gene and / or type II 5α-reductase gene in hair follicles in areas of hair loss enhances the effects of androgens on susceptible follicles. In AGA, cells in the dermis of susceptible follicles contain a specific type II 5α-reductase enzyme that converts testosterone circulating in the area into dihydrotestosterone. This enzyme binds to androgen receptors within these cells, triggering a series of reactions that lead to progressive miniaturization of hair follicles and hair loss, ultimately leading to baldness. Currently, only minoxidil and finasteride are approved by the FDA for the treatment of AGA. Minoxidil is a potassium channel opener, and the exact mechanism of its effectiveness in AGA is not fully understood. It is applied topically to the affected area. However, it has a long onset of action and is associated with adverse reactions such as itching, contact dermatitis, and worsening hair loss after discontinuation of treatment. Finasteride, a type II 5-α-reductase inhibitor, reduces serum and scalp dihydrotestosterone (DHT) levels and is used to treat male AGA. However, long-term use of finasteride can lead to erectile and ejaculatory dysfunction and decreased libido, making it a significant deterrent for men with hair loss. Therefore, research into novel molecules with AR antagonistic activity is crucial for the study of androgenic alopecia.

[0006] Summary of the Invention

[0007] In one aspect, the present disclosure provides a compound represented by formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0008] in,

[0009] X is selected from CH or N;

[0010] Y is selected from OH, COOH, -CONH2, -COOR 3 or -CONR 3’ R 3 ;

[0011] R 1 Selected from OH or -O-C1-C6 alkyl;

[0012] R 2 is selected from H, deuterium, C1-C6 alkyl, C1-C6 deuterated alkyl or C1-C6 haloalkyl;

[0013] R 3 Selected from C1-C6 alkyl, C1-C6 deuterated alkyl, C3-C6 cycloalkyl, 4-10 membered heterocyclic group, C6-C 10Aryl or 5-10 membered heteroaryl, the C1-C6 alkyl, C3-C6 cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally replaced by R 3a replace;

[0014] R 3a Selected from halogen, OH, CN, NH2, -COR 3b 、-COOR 3b 、-NHCOR 3b 、-CONHR 3b , -O-C1-C6 alkyl, C1-C6 alkyl, phenyl, 4-6 membered heterocyclic group or 5-6 membered heteroaryl, wherein the C1-C6 alkyl, phenyl, 4-6 membered heterocyclic group or 5-6 membered heteroaryl is optionally replaced by R 3c replace;

[0015] R 3b Selected from H or C1-C6 alkyl;

[0016] R 3c is selected from halogen, OH, C1-C6 alkyl or -O-C1-C6 alkyl;

[0017] R 3’ Selected from H or C1-C6 alkyl;

[0018] R 4 Selected from NO2, CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl or -O-C1-C6 alkyl;

[0019] R 5 is selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, -COOC1-C6 alkyl, -S-C1-C6 alkyl, -O-C1-C6 alkyl, -Se-C1-C6 alkyl, -S(O)-C1-C6 alkyl or -S(O)2-C1-C6 alkyl;

[0020] R 6 Selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or -O-C1-C6 alkyl;

[0021] n is selected from 1, 2, 3, 4, 5, 6, 7, 8 or 9;

[0022] Provided that, when Y is selected from OH, n is selected from 1, 2 or 3.

[0023] In another aspect, the present disclosure provides a pharmaceutical composition comprising the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0024] In another aspect, the present disclosure provides a method for preventing or treating a disease mediated by androgen receptor in a mammal, comprising administering to a mammal in need of such treatment a therapeutically effective amount of a compound of formula (I) or its stereoisomer or pharmaceutically acceptable salt, or a pharmaceutical composition thereof.

[0025] In another aspect, the present disclosure provides use of a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating androgen receptor-mediated diseases.

[0026] In another aspect, the present disclosure provides a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for use in preventing or treating androgen receptor-mediated diseases.

[0027] In another aspect, the present disclosure provides use of a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in preventing or treating androgen receptor-mediated diseases.

[0028] Detailed description

[0029] The present disclosure provides a compound represented by formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0030] in,

[0031] X is selected from CH or N;

[0032] Y is selected from OH, COOH, -CONH2, -COOR 3 or -CONR 3’ R 3 ;

[0033] R 1 Selected from OH or -O-C1-C6 alkyl;

[0034] R 2 is selected from H, deuterium, C1-C6 alkyl, C1-C6 deuterated alkyl or C1-C6 haloalkyl;

[0035] R 3 Selected from C1-C6 alkyl, C1-C6 deuterated alkyl, C3-C6 cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl, the C1-C6 alkyl, C3-C6 cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally replaced by R 3a replace;

[0036] R 3a Selected from halogen, OH, CN, NH2, -COR3b 、-COOR 3b 、-NHCOR 3b 、-CONHR 3b , -O-C1-C6 alkyl, C1-C6 alkyl, phenyl, 4-6 membered heterocyclic group or 5-6 membered heteroaryl, wherein the C1-C6 alkyl, phenyl, 4-6 membered heterocyclic group or 5-6 membered heteroaryl is optionally replaced by R 3c replace;

[0037] R 3b Selected from H or C1-C6 alkyl;

[0038] R 3c is selected from halogen, OH, C1-C6 alkyl or -O-C1-C6 alkyl;

[0039] R 3’ Selected from H or C1-C6 alkyl;

[0040] R 4 Selected from NO2, CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl or -O-C1-C6 alkyl;

[0041] R 5 is selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, -COOC1-C6 alkyl, -S-C1-C6 alkyl, -O-C1-C6 alkyl, -Se-C1-C6 alkyl, -S(O)-C1-C6 alkyl or -S(O)2-C1-C6 alkyl;

[0042] R 6 Selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or -O-C1-C6 alkyl;

[0043] n is selected from 1, 2, 3, 4, 5, 6, 7, 8 or 9;

[0044] Provided that, when Y is selected from OH, n is selected from 1, 2 or 3.

[0045] In some embodiments, X is N.

[0046] In some embodiments, Y is selected from COOH, -CONH2, -COOR 3 or -CONR 3’ R 3 In some embodiments, Y is selected from -CONH2, -COOR 3 or -CONR 3’ R 3 .

[0047] In some embodiments, Y is selected from -COOR 3 or -CONR3’ R 3 .

[0048] In some embodiments, Y is -COOR 3 .

[0049] In some embodiments, Y is selected from OH, COOH, -COOCH2CH3, -COOCH3, -COOCH(CH3)2, -COOC(CH3)3, -CONHCH3, or -CONH2. In some embodiments, Y is selected from -COOCH2CH3, -COOCH3, -COOCH(CH3)2, -COOC(CH3)3, or -CONHCH3.

[0050] In some embodiments, Y is selected from -COOCH2CH3, -COOCH3, -COOCH(CH3)2, or -COO(CH3)3.

[0051] In some embodiments, R 1 For OH.

[0052] In some embodiments, R 2 Selected from C1-C6 alkyl, C1-C6 deuterated alkyl or C1-C6 haloalkyl.

[0053] In some embodiments, R 2 is selected from H, C1-C6 alkyl or C1-C6 haloalkyl. 2 Selected from H, C1-C3 alkyl or C1-C3 haloalkyl.

[0054] In some embodiments, R 2 is selected from C1-C6 alkyl or C1-C6 haloalkyl. 2 Selected from C1-C3 alkyl or C1-C3 haloalkyl.

[0055] In some embodiments, R 2 Selected from H, CH3 or CF3.

[0056] In some embodiments, R 2 In some embodiments, R 2 Selected from CH3.

[0057] In some embodiments, R 1 OH, R 2 For CH3.

[0058] In some embodiments, R 3Selected from C1-C6 alkyl, C1-C6 deuterated alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclic group, C6-C 10 Aryl or 5-6 membered heteroaryl, the C1-C6 alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclic group, C6-C 10 Aryl or 5-6 membered heteroaryl is optionally replaced by R 3a replace.

[0059] In some embodiments, R 3 is selected from C1-C6 alkyl or C1-C6 deuterated alkyl, wherein the C1-C6 alkyl is optionally replaced by R 3a replace.

[0060] In some embodiments, R 3 In some embodiments, R 3 is a C1-C4 alkyl group, wherein the C1-C4 alkyl group is optionally replaced by R 3a replace.

[0061] In some embodiments, R 3 is an unsubstituted C1-C6 alkyl. 3 It is an unsubstituted C1-C4 alkyl group.

[0062] In some embodiments, R 3 is selected from methyl, ethyl, isopropyl or tert-butyl. 3 is selected from unsubstituted methyl, unsubstituted ethyl, unsubstituted isopropyl or unsubstituted tert-butyl.

[0063] In some embodiments, R 3’ is H or C1-C4 alkyl. In some embodiments, R 3’ For H.

[0064] In some embodiments, R 3a Selected from halogen, OH, CN, NH2, -COR 3b 、-COOR 3b 、-NHCOR 3b 、-CONHR 3b , -O-C1-C6 alkyl or C1-C6 alkyl, the C1-C6 alkyl is optionally replaced by R 3c replace.

[0065] In some embodiments, R 3a is selected from halogen, OH, CN, NH2, -O-C1-C6 alkyl or C1-C6 alkyl, wherein the C1-C6 alkyl is optionally replaced by R 3c replace.

[0066] In some embodiments, R 4 is selected from halogen or CN.

[0067] In some embodiments, R 4 is selected from F, Cl or CN.

[0068] In some embodiments, R 4 For CN.

[0069] In some embodiments, R 5 Selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, -COOC1-C6 alkyl, -S-C1-C6 alkyl or -O-C1-C6 alkyl.

[0070] In some embodiments, R 5 is selected from C1-C6 haloalkyl, -S-C1-C6 alkyl, -O-C1-C6 alkyl, -Se-C1-C6 alkyl, -S(O)-C1-C6 alkyl or -S(O)2-C1-C6 alkyl. In some embodiments, R 5 is selected from C1-C3 haloalkyl, -S-C1-C3 alkyl, -O-C1-C3 alkyl, -Se-C1-C3 alkyl, -S(O)-C1-C3 alkyl or -S(O)2-C1-C3 alkyl. In some embodiments, R 5 Selected from C1-C3 haloalkyl, -S-C1-C3 alkyl, -O-C1-C3 alkyl or -Se-C1-C3 alkyl.

[0071] In some embodiments, R 5 is selected from C1-C6 haloalkyl, -S-C1-C6 alkyl or -O-C1-C6 alkyl. 5 Selected from C1-C3 haloalkyl, -S-C1-C3 alkyl or -O-C1-C3 alkyl.

[0072] In some embodiments, R 5 Selected from CF3, -S-CH3, -O-CH3, -Se-CH3, -S(O)-CH3 or -S(O)2-CH3.

[0073] In some embodiments, R 5 In some embodiments, R 5 In some embodiments, R 5 It is CF3.

[0074] In some embodiments, R 6 is selected from H or halogen.

[0075] In some embodiments, R 6 is selected from H, F or Cl.

[0076] In some embodiments, R 6 For H.

[0077] In some embodiments, n is selected from 1, 2, or 3.

[0078] In some embodiments, n is selected from 1 or 2.

[0079] In some embodiments, n is 2.

[0080] In some embodiments, X is selected from CH or N; Y is selected from OH, COOH, -CONH2, -COOR 3 or -CONR 3’ R 3 , where R 3 is an unsubstituted C1-C6 alkyl group, and R 3’ is H or C1-C6 alkyl; R 1 OH; R 2 is selected from H, C1-C3 alkyl or C1-C3 haloalkyl; R 4 CN; R 5 is selected from C1-C3 haloalkyl, -S-C1-C3 alkyl, -O-C1-C3 alkyl, -Se-C1-C3 alkyl, -S(O)-C1-C3 alkyl or -S(O)2-C1-C3 alkyl; and R 6 For H.

[0081] In some embodiments, X is selected from CH or N; Y is selected from OH, COOH, -COOCH2CH3, -COOCH3, -COOCH(CH3)2, -COOC(CH3)3, -CONHCH3, or -CONH2; R 1 OH; R 2 is selected from H, CH3 or CF3; R 4 CN; R 5 is selected from CF3, -S-CH3, -O-CH3, -Se-CH3, -S(O)-CH3 or -S(O)2-CH3; and R 6 For H.

[0082] In some embodiments, X is selected from CH or N; Y is selected from OH or -COOR 3 , where R 3 is an unsubstituted C1-C6 alkyl group; R 1 OH; R 2 Selected from H or C1-C3 alkyl; R 4 CN; R 5is selected from C1-C3 haloalkyl, -S-C1-C3 alkyl, -O-C1-C3 alkyl, -Se-C1-C3 alkyl; and R 6 For H.

[0083] In some embodiments, X is selected from CH or N; Y is selected from OH, -COOCH2CH3, -COOCH3, -COOCH(CH3)2, or -COOC(CH3)3; R 1 OH; R 2 Selected from H or CH3; R 4 CN; R 5 is selected from CF3, -S-CH3, -O-CH3 or -Se-CH3; and R 6 For H.

[0084] In some embodiments, the compound represented by formula (I) or its stereoisomer or its pharmaceutically acceptable salt is selected from the compound represented by formula (II) or its stereoisomer or its pharmaceutically acceptable salt,

[0085] Among them, X, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 , n are as defined above.

[0086] In some embodiments, the compound represented by formula (I) or its stereoisomer or its pharmaceutically acceptable salt is selected from the following compounds or their stereoisomers or their pharmaceutically acceptable salts:

[0087] In another aspect, the present disclosure provides a pharmaceutical composition comprising the compound of formula (I) or (II) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0088] In another aspect, the present disclosure provides a method for preventing or treating a disease mediated by an androgen receptor in a mammal, comprising administering to a mammal, preferably a human, in need of such treatment, a therapeutically effective amount of a compound of formula (I) or (II) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0089] In another aspect, the present disclosure provides use of a compound of formula (I) or (II) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating androgen receptor-mediated diseases.

[0090] In another aspect, the present disclosure provides use of a compound of formula (I) or (II) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in preventing or treating androgen receptor-mediated diseases.

[0091] In another aspect, the present disclosure provides a compound of formula (I) or (II) or a stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for use in preventing or treating androgen receptor-mediated diseases.

[0092] In some embodiments, the androgen receptor-mediated disease is selected from androgenic alopecia.

[0093] Any embodiment of any aspect of the present disclosure may be combined with other embodiments, provided that no contradiction occurs. In addition, in any embodiment of any aspect of the present disclosure, any technical feature may be applicable to the technical feature in other embodiments, provided that no contradiction occurs.

[0094] Definition and explanation of terms

[0095] Unless otherwise indicated, the terms used in this disclosure have the following meanings. The definitions of groups and terms described in this disclosure, including their definitions as examples, exemplary definitions, preferred definitions, definitions described in tables, and definitions of specific compounds in the examples, may be combined and coupled with each other in any manner. A particular term should not be considered as undefined or unclear unless specifically defined, but should be understood according to its ordinary meaning in the art. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0096] The diagrammatic representations of racemates or enantiomerically pure compounds herein are from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise indicated, wedge and dotted wedge keys are used. To indicate the absolute configuration of a stereocenter, use black real and imaginary bonds. Indicates the relative configuration of a stereocenter (such as the cis-trans configuration of an alicyclic compound).

[0097] The term "tautomer" refers to functional group isomers resulting from the rapid shift of an atom in a molecule between two positions. The compounds of the present disclosure may exhibit tautomerism. Tautomeric compounds may exist as two or more interconvertible species. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer usually result in a mixture whose physical and chemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates, while in phenols, the enol form predominates. The present disclosure encompasses all tautomeric forms of the compounds.

[0098] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0099] The compounds of the present invention may have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms or asymmetric double bonds, so that the compounds of the present invention may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis and trans isomers, E and Z geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures or other mixtures thereof, such as mixtures enriched in enantiomers or diastereomers, all of which are within the definition of the compounds of the present invention and mixtures thereof. Additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms or asymmetric phosphorus atoms may be present in substituents such as alkyl groups, and all of which are within the definition of the compounds of the present invention and mixtures thereof. Compounds of the present disclosure containing an asymmetric atom can be isolated in optically pure or racemic forms. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or reagents.

[0100] Asterisk "*" herein represents a chiral center, indicating that the absolute configuration of the position is either S-configuration or R-configuration.

[0101] The term "substituted" refers to the replacement of any one or more hydrogen atoms on a particular atom by a substituent, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is an oxo (i.e., =O), it means that two hydrogen atoms are replaced. Oxo does not occur on aromatic groups.

[0102] The term "optional" or "optionally" refers to that the event or situation described subsequently may or may not occur, and the description includes that the event or situation occurs and that the event or situation does not occur. For example, an ethyl group is "optionally" substituted with halogen, meaning that the ethyl group may be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl, etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2, etc.), or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3, etc.). It will be appreciated by those skilled in the art that for any group comprising one or more substituents, any sterically impossible and / or incomposable replacement or substitution pattern will not be introduced.

[0103] When any variable (such as R a 、Rb ) appears more than once in the composition or structure of a compound, its definition is independent in each case. For example, if a group is represented by two R b is replaced, then each R b There are independent options.

[0104] When a substituent's bond crosses two atoms in a ring, the substituent can be bonded to any atom in the ring. Indicates that H at any position 1 or 2 on the ring is replaced by R 6 replace.

[0105] In this article Indicates the attachment site.

[0106] In this article, C m -C n It means having an integer number of carbon atoms in the range mn. For example, "C1-C 10 ” means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, or 10 carbon atoms.

[0107] The term "alkyl" refers to a group of the formula C n H 2n+1 The term "C1-C 10 The term "alkyl" is understood to mean a straight-chain or branched saturated hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Specific examples of the alkyl radical include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl , 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc.; the term "C1-C6 alkyl" may be understood to mean an alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms, specific examples of which include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C3 alkyl" may be understood to mean a straight-chain or branched saturated alkyl group having 1, 2 or 3 carbon atoms. The "C1-C 10The term "alkyl" may include "C1-C6 alkyl" or "C1-C3 alkyl" and the like, and the "C1-C6 alkyl" may further include "C1-C3 alkyl".

[0108] The term "deuterated alkyl" refers to a group in which hydrogen on an alkyl group is replaced by deuterium, including monodeuterated alkyl and polydeuterated alkyl. For example, the term "C1-C6 deuterated alkyl" refers to a C1-C6 alkyl group as defined above that is substituted by one or more deuterium groups, including but not limited to CD3, CH2CD3, and the like.

[0109] The term "haloalkyl" is intended to include monohaloalkyl and polyhaloalkyl. For example, the term "C1-C6 haloalkyl" means a C1-C6 alkyl as defined above substituted with one or more halogens, including but not limited to trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, and the like.

[0110] The term "cycloalkyl" refers to a fully saturated carbon ring that exists in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 3- to 10-membered ring. The term "C3-C 10 "Cycloalkyl" is understood to mean a saturated monocyclic, bicyclic, spirocyclic or bridged ring having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Specific examples of the cycloalkyl group include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decyl, and the like. The term "C3-C 10 "Cycloalkyl" may include "C3-C6 cycloalkyl". The term "C3-C6 cycloalkyl" may be understood to mean a saturated monocyclic or bicyclic hydrocarbon ring having 3, 4, 5 or 6 carbon atoms. Specific examples include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0111] The term "heterocyclyl" refers to a fully saturated or partially saturated (heteroaromatic as a whole that is not aromatic) monocyclic, fused, spirocyclic or bridged ring group, which contains 1, 2, 3, 4 or 5 heteroatoms or heteroatomic groups (i.e., heteroatom-containing atomic groups) in its ring atoms, wherein the "heteroatoms or heteroatomic groups" include, but are not limited to, nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(=O)2-, -S(=O)-, -P(=O)2-, -P(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH- or -NHC(=O)NH-. The term "3-10 membered heterocyclyl" refers to a heterocyclyl group having 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms, and containing 1, 2, 3, 4 or 5 heteroatoms or heteroatomic groups independently selected from the above-mentioned heteroatoms or heteroatomic groups in its ring atoms. “3-10 membered heterocyclyl” includes “4-7 membered heterocyclyl”, wherein specific examples of 4 membered heterocyclyl include but are not limited to azetidinyl, thietanyl or oxetanyl; specific examples of 5 membered heterocyclyl include but are not limited to tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl or 2,5-dihydro-1H-pyrrolyl; specific examples of 6 membered heterocyclyl include but are not limited to tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridinyl or 4H-[1,3,4]thiadiazinyl; specific examples of 7 membered heterocyclyl include but are not limited to diazepanyl. The heterocyclic group may also be a bicyclic group, wherein specific examples of 5,5-membered bicyclic groups include but are not limited to hexahydrocyclopenta[c]pyrrol-2(1H)-yl; specific examples of 5,6-membered bicyclic groups include but are not limited to hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group may be a benzo-fused ring group of the above-mentioned 4-7-membered heterocyclic groups, specific examples of which include but are not limited to dihydroisoquinolinyl and the like. "4-10 membered heterocyclyl" may include "5-10 membered heterocyclyl", "4-7 membered heterocyclyl", "5-6 membered heterocyclyl", "6-8 membered heterocyclyl", "4-10 membered heterocycloalkyl", "5-10 membered heterocycloalkyl", "4-7 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", "6-8 membered heterocycloalkyl", etc., and "4-7 membered heterocyclyl" may further include "4-6 membered heterocyclyl", "5-6 membered heterocyclyl", "4-7 membered heterocycloalkyl", "4-6 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", etc. Although some bicyclic heterocyclyl groups in the present disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclyl group as a whole is still non-aromatic.

[0112] The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π electron system. The aryl group may have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. The term "C6-C 20 "Aryl" is understood to be an aromatic radical having 6 to 20 carbon atoms. In particular, a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 or a ring having 13 carbon atoms ("C 13 aryl) such as fluorenyl; or a ring having 14 carbon atoms ("C 14 The term "C6-C 10 "Aryl" is understood to be an aromatic radical having 6 to 10 carbon atoms. In particular, a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 The term "C6-C 20 "Aryl" may contain "C6-C 10 Aryl".

[0113] The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system having aromatic character, which contains at least one ring atom selected from N, O, S, and the remaining ring atoms are C. The term "5-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms, and containing 1, 2, 3, 4 or 5, preferably 1, 2 or 3 heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl, and the like, and benzo derivatives thereof, such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl, and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl or isoquinolyl, and the like; or acinyl, indolizinyl, purinyl, and the like, and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, and the like. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms and containing 1, 2 or 3, preferably 1-2, heteroatoms independently selected from N, O and S.

[0114] The term "halo" or "halogen" refers to fluorine, chlorine, bromine or iodine.

[0115] The term "hydroxy" refers to an -OH group.

[0116] The term "cyano" refers to a -CN group.

[0117] The term "amino" refers to a -NH2 group.

[0118] The term "therapeutically effective amount" means

[0119] An amount of a compound of the present disclosure that (i) treats a particular disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) delays the onset of one or more symptoms of a particular disease, condition, or disorder as described herein.

[0120] The amount of a compound of the disclosure that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one skilled in the art based on their own knowledge and this disclosure.

[0121] The term "treating" means administering a compound or formulation of the present disclosure to improve or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0122] (i) inhibiting a disease or disease state, i.e., arresting its development;

[0123] (ii) ameliorating the disease or condition, i.e., causing regression of the disease or condition.

[0124] The term "preventing" or "preventing" means administering a compound or formulation of the present disclosure to prevent a disease or one or more symptoms associated with the disease, including preventing the disease or disease state from occurring in a mammal, particularly when such mammal is susceptible to the disease state but has not yet been diagnosed as having the disease state.

[0125] In this disclosure, examples of the term "mammal" include, but are not limited to, any member of the class Mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals, such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, etc.

[0126] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0127] The term "pharmaceutically acceptable salt" refers to pharmaceutically acceptable acid addition salts or base addition salts, including salts formed between a compound and an inorganic acid or organic acid, and salts formed between a compound and an inorganic base or an organic base.

[0128] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure or their salts and a pharmaceutically acceptable excipient. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present disclosure to an organism.

[0129] The term "pharmaceutically acceptable excipient" refers to an excipient that is non-irritating to organisms and does not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art and include, for example, carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0130] The word "comprise" or "comprises" and its English variations such as comprises or comprising are to be understood as having an open and non-exclusive meaning, ie, "including but not limited to".

[0131] The present disclosure also includes isotopically labeled compounds of the present disclosure that are identical to those described herein, but where one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0132] Certain isotopically labeled compounds of the present disclosure (e.g.,3 H and 14 C-labeled) can be used in compound and / or substrate tissue distribution assays. 3 H) and carbon-14 (i.e. 14 C) isotopes are particularly preferred due to their ease of preparation and detectability. Positron emitting isotopes, such as 15 O. 13 N. 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the disclosure can generally be prepared by following procedures analogous to those disclosed in the Schemes and / or Examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0133] The pharmaceutical compositions of the present disclosure can be prepared by combining the compounds of the present disclosure with suitable pharmaceutically acceptable excipients, and can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.

[0134] Typical routes of administration of the disclosed compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0135] The pharmaceutical composition of the present disclosure can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, emulsification methods, freeze-drying methods, and the like.

[0136] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present disclosure to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, and the like for oral administration to a patient.

[0137] Solid oral compositions can be prepared by conventional mixing, filling, or tableting methods. For example, they can be prepared by mixing the active compound with a solid excipient, optionally grinding the resulting mixture, adding other suitable excipients as needed, and then granulating the mixture to obtain a tablet or dragee core. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, or flavoring agents.

[0138] The pharmaceutical composition may also be suitable for parenteral administration, such as sterile solutions, suspensions or lyophilized products in appropriate unit dosage forms.

[0139] In all methods of administration of the compounds of formula (I) described herein, the daily dosage is 0.01 mg / kg to 500 mg / kg body weight, in single or divided doses.

[0140] The compounds disclosed herein can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining the same with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples disclosed herein.

[0141] The chemical reactions of the embodiments of the present disclosure are carried out in a suitable solvent that is compatible with the chemical transformations of the present disclosure and the reagents and materials required. In order to obtain the compounds of the present disclosure, it is sometimes necessary for those skilled in the art to modify or select synthetic steps or reaction schemes based on existing embodiments. Example

[0142] The present invention is described in detail below by way of examples, but this is not intended to limit the present disclosure in any way. While the present disclosure has been described in detail herein, including specific embodiments thereof, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present disclosure without departing from the spirit and scope of the present disclosure.

[0143] All reagents used in this disclosure were commercially available and used without further purification.

[0144] Unless otherwise specified, the ratios expressed in mixed solvents are by volume. The eluents described below may be composed of two or more solvents, with the ratios being the volume ratios of the individual solvents. For example, "ethyl acetate / petroleum ether = 5 / 1" indicates that the volume ratio of ethyl acetate to petroleum ether in the mixed eluent during gradient elution is 5:1.

[0145] Unless otherwise stated, % refers to wt%.

[0146] Compounds are manually or The software named the commercially available compounds using the supplier's catalog name.

[0147] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The units of NMR shifts are 10 - 6 (ppm). The solvents for NMR determination are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS); "IC 50 ” refers to the half-maximal inhibitory concentration, which is the concentration at which half of the maximum inhibitory effect is achieved.

[0148] Abbreviations:

[0149] TBAF: tetrabutylammonium fluoride; THF: tetrahydrofuran; TsOH: p-toluenesulfonic acid; BnBr: benzyl bromide; DMF: N,N-dimethylformamide; Cy2NH: dicyclohexylamine; AcCl: acetyl chloride; EtOH: ethanol; EA: ethyl acetate; DMAc: N,N-dimethylacetamide; LiOH . H2O: lithium hydroxide monohydrate; BnOH: benzyl alcohol; DCC: N,N'-dicyclohexylcarbodiimide; DMAP: 4-dimethylaminopyridine; DCM: dichloromethane; ACN: acetonitrile; RuCl3: ruthenium trichloride; MeOH: methanol; i-PrOH: isopropanol; t-BuOH: tert-butanol; TMSCHN2: trimethylsilyldiazomethane; Toluene: toluene; TEA: triethylamine; TMSCF3: (trifluoromethyl)trimethylol silane; CsF: cesium fluoride; Pd2(dba)3: tris(dibenzylideneacetone)dipalladium; dppf: 1,1'-bis(diphenylphosphino)ferrocene; DIEA: N,N-diisopropylethylamine; SOCl2: thionyl chloride; MeMgBr: methylmagnesium bromide; m-CPBA: m-chloroperbenzoic acid; NaBH4: sodium borohydride; EtONa: sodium ethoxide; Acetone: acetone; NaIO4: sodium periodate; DMSO: dimethyl sulfoxide.

[0150] Example 1: Synthesis of N-[6-cyano-5-(trifluoromethyl)pyridin-3-yl]-2,6-dihydroxy-2-methyl-hexanamide (Compound 001)

[0151] Step 1: Synthesis of bromo-[4-[tert-butyl(dimethyl)silyl]oxybutyl]magnesium

[0152] Under nitrogen, magnesium (545 mg, 22.42 mmol) and 4-bromobutoxy-tert-butyl-dimethylsilane (5.0 g, 18.71 mmol) were dissolved in tetrahydrofuran (50 mL). Iodine (1 pellet) was added at 25°C. After complete addition, the reaction mixture was stirred at 70°C for 4 hours. The reaction mixture was cooled to room temperature and used directly in the next reaction.

[0153] Step 2: Synthesis of 6-[tert-butyl(dimethyl)silyl]oxy-N-[6-cyano-5-(trifluoromethyl)pyridin-3-yl]-2-hydroxy-2-methyl-hexanamide

[0154] The reaction mixture from the previous step (33.3 mL) was slowly added dropwise to a solution of N-[6-cyano-5-(trifluoromethyl)pyridin-3-yl]-2-oxo-propionamide (Compound 1-3, 1.0 g, 3.89 mmol) in tetrahydrofuran (20 mL) at -78°C under nitrogen over 15 minutes. The reaction mixture was stirred at -78°C for 1 hour. LCMS indicated the reaction was complete. The reaction mixture was quenched with saturated aqueous ammonium chloride and extracted with ethyl acetate (50 mL x 2). The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (silica, ethyl acetate / petroleum ether = 1 / 1) to afford Compound 1-4 (550 mg).

[0155] MS (ESI): m / z = 446.20 [M+H] + .

[0156] Step 3: Synthesis of N-[6-cyano-5-(trifluoromethyl)pyridin-3-yl]-2,6-dihydroxy-2-methyl-hexanamide

[0157] Tetrabutylammonium fluoride (1.11 mL, 1 M in THF) and intermediate compound 1-4 (450 mg, 1.01 mmol) were dissolved in tetrahydrofuran (15 mL), and the reaction mixture was stirred at 25°C for 1 h. The reaction mixture was concentrated, and the crude product was purified by column chromatography (silica, ethyl acetate / petroleum ether = 1 / 1) to obtain the target compound 001 (115 mg).

[0158] MS (ESI): m / z = 332.10 [M+H] + .

[0159] 1 H NMR (400MHz, DMSO-d6) δ10.70(s,1H),9.40(d,J=2.2Hz,1H),8.92(d,J=2.2Hz,1H),5.85(s,1H),4.33(t,J=5.1Hz,1H) ,3.36(d,J=5.7Hz,2H),1.81–1.69(m,1H),1.67–1.53(m,1H),1.53–1.43(m,1H),1.43–1.32(m,5H),1.28–1.14(m,1H).

[0160] Example 2: Synthesis of N-[6-cyano-5-(trifluoromethyl)pyridin-3-yl]-2,4-dihydroxy-2-methyl-butyramide (Compound 002)

[0161] Step 1: Synthesis of ((1,3-dioxolan-2-yl)methyl)magnesium bromide

[0162] Under nitrogen, add 2-(bromomethyl)-1,3-dioxolane (20 g, 119.76 mmol), lithium chloride (10.15 g, 239.52 mmol), and magnesium powder (3.49 g, 143.71 mmol) to tetrahydrofuran (200 mL). Stir the reaction at 70°C for 4 h. The reaction mixture can be used directly in the next reaction.

[0163] Step 2: Synthesis of N-[6-cyano-5-(trifluoromethyl)pyridin-3-yl]-3-(1,3-dioxolane-2-yl)-2-hydroxy-2-methylpropionamide

[0164] At -78°C under nitrogen, the reaction mixture (200 mL) from the previous step was slowly added dropwise to a solution of N-[6-cyano-5-(trifluoromethyl)pyridin-3-yl]-2-oxo-propionamide (compound 1-3, 2.0 g, 7.78 mmol) in tetrahydrofuran (20 mL). After completion of the addition, the reaction mixture was warmed to 70°C and stirred for 18 hours. LCMS indicated the reaction was complete. The reaction mixture was quenched with saturated aqueous ammonium chloride and extracted with ethyl acetate (50 mL x 2). The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (silica, ethyl acetate / petroleum ether = 1 / 1) to obtain the target compound 2-3 (220 mg).

[0165] MS (ESI): m / z = 346.10 [M+H] + .

[0166] 1 H NMR (400MHz, DMSO-d6) δ10.67(s,1H),9.36(d,J=2.2Hz,1H),8.87(d,J=2.2Hz,1H),6.06(s,1 H),5.06–4.98(m,1H),3.89–3.62(m,4H),2.12–2.05(m,1H),1.98–1.89(m,1H),1.40(s,3H).

[0167] Step 3: Synthesis of N-[6-cyano-5-(trifluoromethyl)pyridin-3-yl]-2-hydroxy-2-methyl-4-oxobutanamide

[0168] Intermediate compound 2-3 (150 mg, 434.44 μmol) and p-toluenesulfonic acid (22.63 mg, 434.44 μmol) were added to tetrahydrofuran (10 mL). The reaction mixture was stirred at room temperature for 18 h. LCMS indicated the reaction was complete. Water (60 mL) and ethyl acetate (80 mL) were added sequentially. The organic phase was washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 2-4 (120 mg).

[0169] MS (ESI): m / z = 302.1 [M+H] + .

[0170] Step 4: Synthesis of N-[6-cyano-5-(trifluoromethyl)pyridin-3-yl]-2,4-dihydroxy-2-methyl-butyramide

[0171] Intermediate compound 2-4 (100 mg, 331.98 μmol) was added to tetrahydrofuran (1 mL). Sodium borohydride (18.84 mg, 497.97 μmol) was added under ice-water bath. The reaction mixture was stirred under ice-water bath for 18 h. The mixture was quenched by adding 2 mL of water under ice-water bath. The reaction mixture was concentrated, and the residue was purified by Prep-HPLC (Phenomenex C18, 5 μm silica, 30 mm diameter, 80 mm length; eluents were composed of decreasingly polar mixtures of water (containing 7 mmol / L NH4HCO3) and acetonitrile; acetonitrile gradient ratio 60%-70%, elution time: 9 minutes) to afford the target compound 002 (8.2 mg).

[0172] MS (ESI): m / z = 304.10 [M+H] + .

[0173] 1 H NMR(400MHz,DMSO-d6)δ9.36(d,J=2.2Hz,1H),8.89(d,J=2.2Hz,1H),6.05(s ,1H),3.62–3.43(m,2H),2.05–1.93(m,1H),1.85–1.73(m,1H),1.37(s,3H).

[0174] Example 3: Synthesis of ethyl 5-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-4-hydroxy-4-methyl-5-oxopentanoate (Compound 003)

[0175] Step 1: Synthesis of 5-(benzyloxy)-4,5-dioxopentanoic acid

[0176] 2-Ketoglutaric acid (2 g, 13.69 mmol) was dissolved in N,N-dimethylformamide (80 mL). Dicyclohexylamine (1.24 g, 6.84 mmol, 1.36 mL) and benzyl bromide (1.17 g, 6.84 mmol) were added sequentially. The mixture was heated to 50°C and stirred overnight. LC-MS indicated the reaction was complete. The reaction solution was poured into methyl tert-butyl ether (300 mL) and washed three times with water (100 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 1 / 2) to afford compound 3-2 (1.1 g).

[0177] MS (ESI): m / z = 235.00 [MH] - .

[0178] 1 H NMR (400MHz, DMSO-d6) δ12.27(s,1H),7.48–7.32(m,5H),5.26(s,2H),3.09-3.01(m,2H),2.52–2.46(m,2H).

[0179] Step 2: Synthesis of ethyl 5-(benzyloxy)-4,5-dioxopentanoate

[0180] Intermediate compound 3-2 (1 g, 4.23 mmol) was dissolved in anhydrous ethanol (10 mL). Acetyl chloride (365 mg, 4.66 mmol) was added in an ice-water bath. The mixture was incubated and stirred for 3 h. LC-MS indicated the reaction was complete. The reaction solution was diluted with ethyl acetate (100 mL) and washed three times with saturated sodium bicarbonate solution (50 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 1 / 5) to afford compound 3-3 (700 mg).

[0181] MS (ESI): m / z = 282.1 [M + NH4] + .

[0182] 1 H NMR (400MHz, Chloroform-d) δ7.45–7.25(m,5H),5.30(s,2H),4.20–4.05(m,2H),3.20–3.12(m,2H),2.70–2.62(m,2H),1.29–1.21(m,3H).

[0183] Step 3: Synthesis of 5-ethoxy-2,5-dioxopentanoic acid

[0184] Intermediate compound 3-3 (500 mg, 1.89 mmol) was dissolved in ethyl acetate (10 mL), and Pd / C (46 mg) was added. The mixture was stirred under a hydrogen atmosphere for 15 minutes. TLC indicated the reaction was complete (developing solvent: dichloromethane / methanol = 10 / 1, Rf = 0.4). The reaction mixture was filtered, and the filtrate was concentrated to obtain compound 3-4 (300 mg).

[0185] 1 H NMR (400MHz, Chloroform-d) δ4.2–4.08(m,2H),3.26–3.12(m,2H),2.77–2.62(m, 2H),1.31–1.20(m,3H).

[0186] Step 4: Synthesis of ethyl 5-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-4,5-dioxopentanoate

[0187] Intermediate compound 3-4 (300 mg, 2.05 mmol) was dissolved in N,N-dimethylacetamide (12 mL), cooled to -10°C, and thionyl chloride (244.22 mg, 2.05 mmol) was added. The mixture was stirred for 10 minutes, and 5-amino-3-(trifluoromethyl)pyridine-2-carbonitrile (384 mg, 2.05 mmol) was added. The mixture was kept warm and stirred for 3 hours. LC-MS indicated that the reaction was complete. The reaction solution was poured into ice water (100 mL) and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was isolated and purified by column chromatography (silica, ethyl acetate / petroleum ether = 1 / 2) to obtain compound 3-5 (300 mg).

[0188] MS (ESI): m / z = 344.0 [M+H] + .

[0189] 1 H NMR (400MHz, DMSO-d6) δ10.73(s,1H),9.40(d,J=2.2Hz,1H),8.91(d,J=2.3Hz,1 H),4.11–4.02(m,2H),3.32–3.16(m,2H),2.68–2.60(m,2H),1.25–1.15(m,3H).

[0190] Step 5: Synthesis of ethyl 5-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-4-hydroxy-4-methyl-5-oxopentanoate

[0191] Intermediate compound 3-5 (100 mg, 291.32 μmol) was dissolved in tetrahydrofuran (10 mL), cooled to -78°C, and methylmagnesium bromide (0.5 M in THF, 1.40 mL) was added dropwise. The mixture was incubated for 2 h, then slowly warmed to room temperature and stirred overnight. LC-MS indicated the reaction was complete. The reaction solution was poured into ice water (40 mL) and extracted with ethyl acetate (15 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The residue was purified by Prep-HPLC using a YMC-TAR column (5 μm silica, 30 mm diameter, 150 mm length); eluents were a decreasingly polar mixture of water (containing 7 mmol / L NH₄HCO₃) and acetonitrile; the acetonitrile gradient was 45% to 75% over 9 minutes) to afford the target compound 003 (15 mg).

[0192] MS (ESI): m / z = 360.10 [M+H] + .

[0193] 1 H NMR(400MHz,DMSO-d6)δ9.37(d,J=2.3Hz,1H),8.89(d,J=2.3Hz,1H),6.01(s,1H),4.06–3.95(m,2H),2.4 7–2.39(m,1H),2.29–2.18(m,1H),2.10–1.97(m,1H),1.94–1.81(m,1H),1.37(s,3H),1.18–1.10(m,3H).

[0194] Example 4: Synthesis of 5-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-4-hydroxy-4-methyl-5-oxopentanoic acid (Compound 004)

[0195] Compound 003 (80 mg, 222.65 μmol) was dispersed in a mixture of tetrahydrofuran (3 mL) and water (1 mL), and lithium hydroxide monohydrate (8.55 mg, 203.76 μmol) was added and stirred at room temperature for 3 h. LC-MS showed that the reaction was complete. The reaction solution was poured into 0.5 M hydrochloric acid solution (15 mL) and extracted three times with ethyl acetate (5 mL*3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by RP-FLASH [Chromatographic column: 40 g C18 Column, 20-35 μm, Mobile phase A: pure water, mobile phase B: acetonitrile; flow rate: 18 mL / min; gradient: 30% B to 50% B, elution time: 10 minutes] to obtain the target compound 004 (15 mg).

[0196] MS (ESI): m / z = 332.0 [M+H] + .

[0197] 1 H NMR(400MHz,DMSO-d6)δ10.71(s,1H),9.38(d,J=2.2Hz,1H),8.92–8.88(m,1H),2.43 –2.32(m,1H),2.22–2.12(m,1H),2.06–1.93(m,1H),1.91–1.80(m,1H),1.37(s,3H).

[0198] Examples 5 & 6: Synthesis of (6S) / (6R)-6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid ethyl ester (Compounds 005 & 006)

[0199] Step 1: Synthesis of benzyl cyclopent-1-ene-1-carboxylate

[0200] Starting material 5-1 (8 g, 71.35 mmol) was dissolved in dichloromethane (200 mL). Benzyl alcohol (9.26 g, 85.62 mmol), N,N'-dicyclohexylcarbodiimide (17.67 g, 85.62 mmol), and 4-dimethylaminopyridine (871.64 mg, 7.13 mmol) were added and stirred at room temperature overnight. TLC (developing solvent: ethyl acetate / petroleum ether = 1 / 3, Rf = 0.6) indicated the reaction was complete. The reaction solution was concentrated, and the residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 0 / 1 to 1 / 4) to obtain the title compound 5-2 (13 g).

[0201] 1 H NMR (400MHz, Chloroform-d) δ7.42–7.27(m,5H),6.87–6.79(m,1H),5.19(s,2H),2.65–2.55(m,2H),2.55–2.45(m,2H),2.02–1.90(m,2H).

[0202] Step 2: Synthesis of 6-(benzyloxy)-5,6-dioxohexanoic acid

[0203] Intermediate compound 5-2 (5 g, 24.72 mmol) was dissolved in a mixture of carbon tetrachloride (120 mL) and acetonitrile (120 mL). A solution of sodium periodate (10.58 g, 49.44 mmol) in water (120 mL) was added dropwise at 0°C. Ruthenium trichloride (512.79 mg, 2.47 mmol) was then added as a solid. The mixture was slowly warmed to room temperature and stirred for 6 h. LC-MS indicated completion of the reaction. The reaction mixture was poured into water (300 mL) and extracted with dichloromethane (150 mL x 3). The organic phases were combined, washed three times with water (100 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The residue was purified by column chromatography (silica, ethyl acetate:petroleum ether = 0 / 1 to 1 / 1) to yield the title compound 5-3 (2.5 g).

[0204] MS (ESI): m / z = 249.10 [MH] - .

[0205] 1 H NMR (400MHz, Chloroform-d) δ7.44–7.31(m,5H),5.28(s,2H),2.99–2.88(m,2H),2.47–2.38(m,2H),2.07–1.90(m,2H).

[0206] Step 3: Synthesis of 1-benzyl-6-ethyl 2-oxoadipate

[0207] Intermediate compound 5-3 (1 g, 4.00 mmol) was dissolved in anhydrous ethanol (10 mL). Acetyl chloride (378.83 mg, 4.82 mmol) was added dropwise in an ice-water bath. The mixture was incubated and the reaction continued for 5 h. LC-MS indicated the reaction was complete. The reaction solution was poured into dichloromethane (30 mL), washed twice with water (10 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 0 / 1 to 1 / 1) to obtain the title compound 5-4 (800 mg).

[0208] MS (ESI): m / z = 279.10 [M+H] + .

[0209] 1 H NMR(400MHz,Chloroform-d)δ7.44–7.31(m,5H),5.28(s,2H),4.17–4.07(m,2H) ),2.97–2.89(m,2H),2.42–2.32(m,2H),2.03–1.90(m,2H),1.41–1.20(m,3H).

[0210] Step 4: Synthesis of 6-ethoxy-2,6-dioxohexanoic acid

[0211] Intermediate compound 5-4 (350 mg, 1.26 mmol) was dissolved in ethyl acetate (4 mL). Pd / C (30 mg) was added and the reaction was stirred under a hydrogen atmosphere for 15 min. TLC (developing solvent: methanol / dichloromethane = 1 / 10, Rf = 0.3) indicated complete reaction. Filter and concentrate the filtrate to obtain the title compound 5-5 (220 mg).

[0212] 1 H NMR (400MHz, Chloroform-d) δ4.19–4.09(m,2H),3.07–2.99(m,2H),2.44–2.35(m,2H),2.07–1.94(m,2H),1.30–1.22(m,3H).

[0213] Step 5: Synthesis of ethyl 6-(6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5,6-dioxohexanoate

[0214] Intermediate compound 5-5 (360 mg, 1.91 mmol) was dissolved in N,N-dimethylacetamide (7.97 mL), cooled to -10°C, and thionyl chloride (273.12 mg, 2.30 mmol) was added dropwise. 5-amino-3-(trifluoromethyl)pyridine-2-carbonitrile (357.98 mg, 1.91 mmol) was added as a solid, and the mixture was stirred at this temperature for 2 h. LC-MS indicated the reaction was complete. The reaction solution was diluted with ethyl acetate (50 mL), washed three times with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 0 / 1 to 1 / 1) to afford the title compound 5-6 (450 mg).

[0215] MS (ESI): m / z = 358.10 [M+H] + .

[0216] 1 H NMR (400MHz, DMSO-d6) δ11.43(s,1H),9.39(d,J=2.3Hz,1H),8.85(d,J=2.3Hz,1H),4.11– 3.98(m,2H),3.02–2.94(m,2H),2.42–2.33(m,2H),1.88–1.76(m,2H),1.21–1.15(m,3H).

[0217] Step 6: Synthesis of ethyl 6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoate

[0218] Intermediate 5-6 was prepared on a scaled-up basis using the method described in Steps 1-5. Intermediate compound 5-6 (17 g, 47.58 mmol) was dissolved in tetrahydrofuran (200 mL) and cooled to -78°C under a nitrogen atmosphere. A 3 M solution of methylmagnesium bromide in tetrahydrofuran (47.58 mL) was added dropwise, and the mixture was stirred overnight. LC-MS indicated the reaction was complete. At -78°C, saturated aqueous ammonium chloride (350 mL) was slowly added to quench the reaction. The mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 0 / 1 to 1 / 1) to afford the title compound 5-7 (11 g).

[0219] MS (ESI): m / z = 374.10 [M+H] + .

[0220] 1 H NMR (400MHz, DMSO-d6) δ10.72(s,1H),9.40(d,J=2.2Hz,1H),8.91(d,J=2.3Hz,1H),5.92(s,1H),4.07– 3.97(m,2H),2.31–2.22(m,2H),1.81–1.65(m,3H),1.50–1.38(m,1H),1.36(s,3H),1.20–1.11(m,3H).

[0221] Step 7: Synthesis of (6S) / (6R)-ethyl 6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoate

[0222] Compound 5-7 (1 g) was separated by SFC [chromatographic column: Chiral Cel OJ-H 150×4.6 mm ID, 5 μm; mobile phase: A: carbon dioxide; B: methanol (0.05% diethanolamine), gradient: mobile phase B: 5% to 40%, elution 4.5 minutes; mobile phase B: 5%, elution 1.5 minutes; flow rate: 2.5 mL / min; column temperature: 40°C] to give compound 005 (310 mg) and compound 006 (330 mg):

[0223] Compound 005:

[0224] SFC retention time: 2.247 min.

[0225] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C] Retention time: 24.980 minutes.

[0226] MS (ESI): m / z = 374.10 [M+H] + .

[0227] 1 H NMR (400MHz, DMSO-d6) δ10.72(s,1H),9.40(d,J=2.2Hz,1H),8.91(d,J=2.3Hz,1H),5.92(s,1H),4.07– 3.97(m,2H),2.31–2.22(m,2H),1.81–1.65(m,3H),1.50–1.38(m,1H),1.36(s,3H),1.20–1.11(m,3H).

[0228] Compound 006:

[0229] SFC retention time: 3.064 min.

[0230] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C] Retention time: 16.224 minutes.

[0231] MS (ESI): m / z = 374.10 [M+H] + .

[0232] 1 H NMR (400MHz, DMSO-d6) δ10.72(s,1H),9.40(d,J=2.2Hz,1H),8.91(d,J=2.3Hz,1H),5.92(s,1H),4.07– 3.97(m,2H),2.31–2.22(m,2H),1.81–1.65(m,3H),1.50–1.38(m,1H),1.36(s,3H),1.20–1.11(m,3H).

[0233] Examples 7 & 8: Synthesis of (6S) / (6R)-6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid methyl ester (Compounds 007 & 008)

[0234] Step 1: Synthesis of (6S) / (6R)-6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid

[0235] Disperse compound 005 (or 006) (270 mg, 723.23 μmol) in a mixture of tetrahydrofuran (3 mL) and water (1 mL). Add lithium hydroxide monohydrate (34.66 mg, 826.02 μmol) and stir at room temperature overnight. LC-MS indicates the reaction is complete. Pour the reaction solution into 0.1 M hydrochloric acid solution (15 mL) and extract with ethyl acetate (5 mL x 3). Combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain compound 7-1 (or 8-1) (240 mg).

[0236] Compound 7-1:

[0237] MS (ESI): m / z = 346.10 [M+H] + .

[0238] 1 H NMR (400MHz, DMSO-d6) δ11.97(s,1H),10.70(s,1H),9.40(d,J=2.2Hz,1H),8.91(d,J=2.3Hz ,1H),5.90(s,1H),2.19(t,J=7.1Hz,2H),1.81–1.54(m,3H),1.48–1.37(m,1H),1.37(s,3H).

[0239] Compound 8-1:

[0240] MS (ESI): m / z = 346.10 [M+H] + .

[0241] 1 H NMR (400MHz, DMSO-d6) δ11.97(s,1H),10.69(s,1H),9.39(d,J=2.2Hz,1H),8.91(d,J=2.2Hz ,1H),5.90(s,1H),2.19(t,J=7.1Hz,2H),1.82–1.54(m,3H),1.48–1.38(m,1H),1.36(s,3H).

[0242] Step 2: Synthesis of methyl (6S) / (6R)-6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoate

[0243] Intermediate compound 7-1 (or 8-1) (50 mg, 144.81 μmol) was dissolved in methanol (4 mL). Thionyl chloride (51.69 mg, 434.44 μmol) was added dropwise at room temperature. The temperature was raised to 60°C and the reaction was stirred for 2 h. LC-MS indicated the reaction was complete. The solvent was evaporated under reduced pressure, and the residue was purified by Prep-HPLC (YMC-TAR column, 5 μm silica, 30 mm diameter, 150 mm length; eluent: decreasingly polar mixtures of water (containing 7 mmol / L NH4HCO3) and acetonitrile; acetonitrile gradient: 40% to 70% over 9.5 minutes) to afford target compound 007 (12 mg) (or 008 (15 mg)).

[0244] Compound 007:

[0245] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C] Retention time: 23.394 minutes.

[0246] MS (ESI): m / z = 360.10 [M+H] + .

[0247] 1 H NMR (400MHz, DMSO-d6) δ10.71(s,1H),9.40(d,J=2.2Hz,1H),8.91(d,J=2.2Hz,1H),5.91( s,1H),3.56(s,3H),2.33–2.25(m,2H),1.83–1.55(m,3H),1.51–1.39(m,1H),1.36(s,3H).

[0248] Compound 008:

[0249] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150 × 4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: Mobile phase B: 10% to 90%, elution over 35 minutes; Flow rate: 1.0 mL / min; Column temperature: 25°C] Retention time: 15.201 min.

[0250] MS (ESI): m / z = 360.10 [M+H] + .

[0251] 1H NMR (400MHz, DMSO-d6) δ10.73(s,1H),9.40(d,J=2.2Hz,1H),8.91(d,J=2.3Hz,1H),5.94( s,1H),3.56(s,3H),2.32–2.24(m,2H),1.81–1.53(m,3H),1.50–1.38(m,1H),1.36(s,3H).

[0252] Examples 9 & 10: Synthesis of (6S) / (6R)-6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid isopropyl ester (Compounds 009 & 010)

[0253] Intermediate compound 7-1 (or 8-1) (50 mg, 144.81 μmol) was dissolved in isopropanol (4 mL). Thionyl chloride (52 mg, 434.44 μmol) was added dropwise at room temperature. The temperature was raised to 60°C and the reaction was stirred for 2 h. LC-MS indicated the reaction was complete. The solvent was evaporated under reduced pressure, and the residue was purified by Prep-HPLC (YMC-TAR column, 5 μm silica, 30 mm diameter, 150 mm length; eluent: decreasingly polar mixtures of water (containing 7 mmol / L NH4HCO3) and acetonitrile; acetonitrile gradient ratio 45%-75% over 11 minutes) to afford the target compound 009 (10 mg) (or 010 (12 mg)).

[0254] Compound 009:

[0255] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C] Retention time: 26.011 minutes.

[0256] MS (ESI): m / z = 388.10 [M+H] + .

[0257] 1 H NMR (400MHz, DMSO-d6) δ10.71(s,1H),9.39(d,J=2.2Hz,1H),8.91(d,J=2.3Hz,1H),5.91(s,1H),4.93– 4.79(m,1H),2.28–2.17(m,2H),1.83–1.53(m,3H),1.50–1.38(m,1H),1.36(s,3H),1.18–1.12(m,6H).

[0258] Compound 010:

[0259] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C] Retention time: 16.601 min.

[0260] MS (ESI): m / z = 388.10 [M+H] + .

[0261] 1 H NMR (400MHz, DMSO-d6) δ10.71(s,1H),9.39(d,J=2.2Hz,1H),8.91(d,J=2.3Hz,1H),5.91(s,1H),4.93– 4.80(m,1H),2.28–2.17(m,2H),1.81–1.54(m,3H),1.49–1.39(m,1H),1.36(s,3H),1.18–1.11(m,6H).

[0262] Examples 11 & 12: Synthesis of (6S) / (6R)-tert-butyl 6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoate (Compounds 011 & 012)

[0263] Intermediate compound 7-1 (or 8-1) (300.00 mg, 868.88 μmol) and O-tert-butyl-N,N'-diisopropylisourea (1.74 g, 8.69 mmol) were added to dichloromethane (7.5 mL) and tert-butanol (644.01 mg, 8.69 mmol) and stirred at room temperature for 15 h. LCMS indicated the reaction was complete. The reaction solution was filtered, the filtrate was concentrated, and the residue was purified by Prep-HPLC [Phenomenex C18, 5 μm silica, 30 mm diameter, 80 mm length; using a mixture of pure water and acetonitrile (acetonitrile content: 60%-70%) as the eluent, elution time: 8 minutes] to obtain the target compound 011 (150 mg) (or 012 (110 mg)).

[0264] Compound 011:

[0265] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C] Retention time: 32.009 minutes.

[0266] MS (ESI): m / z = 400.1 [MH] - .

[0267] 1 H NMR (400MHz, DMSO-d6) δ10.70(s,1H),9.39(s,1H),9.01–8.88(m,1H),5.90(s,1H),2.22–2.13(m,2H),1.82–1.48(m,4H),1.44–1.32(m,12H).

[0268] Compound 012:

[0269] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C] Retention time: 17.147 minutes.

[0270] MS (ESI): m / z = 400.1 [MH] - .

[0271] 1 H NMR (400MHz, DMSO-d6) δ10.70(s,1H),9.39(s,1H),9.00–8.84(m,1H),5.89(s,1H),2.22–2.11(m,2H),1.83–1.49(m,4H),1.46–1.32(m,12H).

[0272] Examples 13 & 14: Synthesis of (6S) / (6R)-6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid ethyl ester (Compounds 013 & 014)

[0273] Step 1: Synthesis of 6-bromo-5-fluoropyridin-3-amine

[0274] 5-Fluoropyridin-3-amine (13.0 g, 116 mmol) was dissolved in N,N-dimethylformamide (60 mL) and cooled in an ice bath. A solution of N-bromosuccinimide (20.64 g, 116 mmol) in N,N-dimethylformamide (50 mL) was slowly added dropwise while still in an ice bath. The reaction was continued in an ice bath for 30 minutes. LC-MS monitoring was performed until the reaction was complete. The reaction solution was poured into 500 mL of water and extracted with ethyl acetate (200 mL x 3). The combined organic phases were washed sequentially with water (300 mL x 2) and saturated brine (300 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a brownish-yellow crude product, which was purified by column chromatography (silica, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to afford Intermediate 13-2 (8.4 g).

[0275] MS(ESI):m / z=190.90,192.90[M+H] + .

[0276] Step 2: Synthesis of 5-amino-3-fluoropyridine-2-carbonitrile

[0277] Intermediate 13-2 (6 g, 31.41 mmol) and cuprous cyanide (11.25 g, 125.64 mmol) were dispersed in N,N-dimethylformamide (100 mL). The mixture was then equipped with a nitrogen balloon and the atmosphere was replaced three times. The mixture was then placed in a 130°C oil bath under a protective atmosphere and reacted for 6 hours. LC-MS indicated the reaction was complete. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (silica, petroleum ether / ethyl acetate = 1 / 2) to afford the title compound 13-3 (2.6 g).

[0278] MS (ESI): m / z = 138.0 [M+H] + .

[0279] Step 3: Synthesis of 5-amino-3-(methylthio)pyridine-2-carbonitrile

[0280] Compound 13-3 (2 g, 14.59 mmol) was added to a reaction flask and dissolved in N,N-dimethylformamide (30 mL). Sodium thiomethoxide (2.04 g, 29.11 mmol) was added with stirring at room temperature. The resulting reaction solution was stirred at room temperature for 12 hours. The reaction solution was poured into water (200 mL) and extracted with ethyl acetate (80 mL*5). The organic phases were combined and washed with water (200 mL*2) and saturated brine (400 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound 13-4 (1.7 g).

[0281] MS (ESI): m / z = 166.0 [M+H]+ .

[0282] 1 H NMR (400MHz, DMSO-d6) δ7.75 (d, J = 2.3Hz, 1H), 6.84 (d, J = 2.3Hz, 1H), 6.47 (s, 2H), 2.50 (s, 3H).

[0283] Step 4: Synthesis of ethyl 6-(6-cyano-5-(methylthio)pyridin-3-yl)amino)-5,6-dioxohexanoate

[0284] Intermediate compound 5-5 (1.37 g, 7.28 mmol) was dissolved in N,N-dimethylacetamide (15 mL), cooled to -10°C, and thionyl chloride (1.04 g, 8.72 mmol) was added dropwise. After completion of the dropwise addition, solid compound 13-4 (1.2 g, 7.26 mmol) was added and stirred for 12 h. LC-MS indicated a viable reaction. The mixture was diluted with ethyl acetate (30 mL), washed three times with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The residue was isolated and purified by column chromatography (silica, ethyl acetate:petroleum ether = 0 / 1 to 1 / 1) to afford compound 13-5 (1.6 g).

[0285] MS (ESI): m / z = 336.0 [M+H] + .

[0286] 1 H NMR(400MHz,Chloroform-d)δ8.96(s,1H),8.41(s,2H),4.19–4.07(m,2H),3.13–3 .04(m,2H),2.60(s,3H),2.46–2.38(m,2H),2.09–1.96(m,2H),1.31–1.22(m,3H).

[0287] Step 5: Synthesis of ethyl 6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoate

[0288] Intermediate compound 13-5 (1 g, 2.98 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to -78°C, and methylmagnesium bromide (3 M in Et2O, 2.98 mL) was added dropwise. The temperature was slowly raised to room temperature and stirred overnight. LC-MS indicated a viable reaction. At 0°C, saturated aqueous ammonium chloride (50 mL) was added to quench the reaction. The reaction was returned to room temperature and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The residue was purified by column chromatography (silica, ethyl acetate:petroleum ether = 1 / 4) to afford compound 13-6 (600 mg).

[0289] MS (ESI): m / z = 352.0 [M+H] + .

[0290] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.95(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H),5.86(s,1H),4.08–3.97(m,2H),2.58( s,3H),2.26(t,J=7.1Hz,2H),1.80–1.66(m,2H),1.64–1.53(m,1H),1.44(d,J=7.6Hz,1H),1.35(s,3H),1.20–1.11(m,3H).

[0291] Step 6: Synthesis of (6S) / (6R)-ethyl 6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoate

[0292] Compound 13-6 (1 g) was separated by SFC [chromatographic column: Chiral Cel OJ-H 150×4.6 mm ID, 5 μm; mobile phase: A: carbon dioxide; B: ethanol (0.05% diethanolamine), gradient: mobile phase B: 5% to 40%, elution in 4.5 minutes; mobile phase B: 5%, elution in 1.5 minutes; flow rate: 2.5 mL / min; column temperature: 40°C] to give compound 013 (350 mg) and compound 014 (370 mg).

[0293] Compound 013:

[0294] SFC retention time: 3.468 min.

[0295] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C] Retention time: 21.615 minutes.

[0296] MS (ESI): m / z = 352.00 [M+H] + .

[0297] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.95(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H),5.86(s,1H),4.08–3.97(m,2H),2. 58(s,3H),2.31–2.22(m,2H),1.80–1.66(m,2H),1.64–1.53(m,1H),1.47–1.40(m,1H),1.35(s,3H),1.20–1.11(m,3H).

[0298] Compound 014:

[0299] SFC retention time: 4.292 min.

[0300] Chiral-HPLC [Chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); Mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C] Retention time: 15.077 minutes.

[0301] MS (ESI): m / z = 352.00 [M+H] + .

[0302] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.95(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H),5.86(s,1H),4.08–3.97(m,2H),2. 58(s,3H),2.31–2.22(m,2H),1.80–1.66(m,2H),1.64–1.53(m,1H),1.47–1.40(m,1H),1.35(s,3H),1.20–1.11(m,3H).

[0303] Examples 15 & 16: Synthesis of (6S) / (6R)-6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid methyl ester (Compounds 015 & 016)

[0304] Step 1: Synthesis of (6S) / (6R)-6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid

[0305] Compound 013 (or Compound 014) (100 mg, 284.56 μmol) was dispersed in a mixture of tetrahydrofuran (3 mL) and water (1 mL). Lithium hydroxide monohydrate (35.82 mg, 853.68 μmol) was added and stirred at room temperature for 3 h. LC-MS indicated the reaction was complete. The reaction solution was poured into 0.1 M hydrochloric acid solution (15 mL) and extracted with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield Compound 15-1 (90 mg) (or 16-1).

[0306] Compound 15-1:

[0307] MS (ESI): m / z = 324.10 [M+H] + .

[0308] 1 H NMR (400MHz, DMSO-d6) δ11.98(s,1H),10.31(s,1H),8.95(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H),5 .84(s,1H),2.58(s,3H),2.19(t,J=7.1Hz,2H),1.81–1.52(m,3H),1.46–1.37(m,1H),1.35(s,3H).

[0309] Compound 16-1:

[0310] MS (ESI): m / z = 324.10 [M+H] + .

[0311] 1H NMR (400MHz, DMSO-d6) δ11.98(s,1H),10.31(s,1H),8.95(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H),5 .84(s,1H),2.58(s,3H),2.19(t,J=7.1Hz,2H),1.81–1.52(m,2H),1.47–1.38(m,1H),1.35(s,3H).

[0312] Step 2: Synthesis of methyl (6S) / (6R)-6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoate

[0313] Compound 15-1 (or 16-1) (90 mg, 278.32 μmol) was dissolved in a mixture of toluene (4 mL) and methanol (1 mL). The mixture was cooled to 0°C and trimethylsilyldiazomethane (2 M in n-hexane, 278.32 μL) was added dropwise. The mixture was stirred for 2 h. LC-MS indicated the reaction was complete. Two drops of acetic acid were added to quench the reaction. The reaction solution was diluted with ethyl acetate (20 mL) and washed three times with water (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 1 / 1) to yield compound 015 (65 mg) (or 016 (65 mg)).

[0314] Compound 015:

[0315] Chiral-HPLC retention time: 19.582 min.

[0316] MS (ESI): m / z = 338.0 [M+H] + .

[0317] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.95(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H),5.86(s,1H),3.56(s,3 H),2.58(s,3H),2.33–2.22(m,2H),1.81–1.64(m,2H),1.64–1.52(m,1H),1.48–1.38(m,1H),1.35(s,3H).

[0318] Compound 016:

[0319] Chiral-HPLC retention time: 13.855 min.

[0320] MS (ESI): m / z = 338.0 [M+H] + .

[0321] 1 H NMR (400MHz, DMSO-d6) δ10.31(s,1H),8.95(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H),5.86(s,1H),3.56(s,3 H),2.58(s,3H),2.33–2.24(m,2H),1.81–1.63(m,2H),1.63–1.53(m,1H),1.52–1.38(m,1H),1.35(s,3H).

[0322] Examples 17 & 18: Synthesis of (6S) / (6R)-6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid isopropyl ester (Compounds 017 & 018)

[0323] Intermediate compound 15-1 (or 16-1) (140 mg, 432.94 μmol) was added to isopropanol (4 mL), and thionyl chloride (51.51 mg, 432.94 μmol) was added dropwise. The temperature was raised to 60°C and stirred for 4 h. LC-MS indicated the reaction was complete. The reaction solution was cooled to room temperature, diluted with ethyl acetate (10 mL*3), and washed three times with water (10 mL*3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by column chromatography (silica, petroleum ether / ethyl acetate = 1 / 1) to obtain compound 017 (130 mg) (or 018 (125 mg)).

[0324] Compound 017:

[0325] Chiral-HPLC [chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C], retention time: 22.544 min.

[0326] MS (ESI): m / z = 366.10 [M+H] + .

[0327] 1H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.95(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H),5.85(s,1H),4.91–4.80(m,1H),2. 58(s,3H),2.27–2.18(m,2H),1.81–1.63(m,2H),1.63–1.52(m,1H),1.50–1.37(m,1H),1.35(s,3H),1.19–1.12(m,6H).

[0328] Compound 18:

[0329] Chiral-HPLC [chromatographic column: AD-RH (Dacelide 150×4.6 mm, 5 μm); mobile phase: A: water; B: acetonitrile, gradient: mobile phase B: 10% to 90%, elution over 35 minutes; flow rate: 1.0 mL / min; column temperature: 25°C], retention time: 15.406 minutes.

[0330] MS (ESI): m / z = 366.10 [M+H] + .

[0331] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.95(d,J=2.0Hz,1H),8.41(d,J=2.1Hz,1H),5.87(s,1H),4.92–4.79(m,1H),2. 58(s,3H),2.27–2.19(m,2H),1.81–1.66(m,2H),1.64–1.53(m,1H),1.49–1.40(m,1H),1.35(s,3H),1.19–1.12(m,6H).

[0332] Example 19: Synthesis of tert-butyl 6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoate (Compound 019)

[0333] Step 1: Synthesis of 6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid

[0334] Intermediate compound 13-6 (300 mg, 853.68 μmol) was dispersed in a mixture of tetrahydrofuran (4 mL) and water (1 mL). Lithium hydroxide monohydrate (38.51 mg, 917.78 μmol) was added and stirred at room temperature overnight. LC-MS indicated the reaction was complete. The reaction solution was poured into 0.1 M hydrochloric acid solution (15 mL) and extracted with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the title compound 19-1 (253 mg).

[0335] MS (ESI): m / z = 324.10 [M+H] + .

[0336] 1 H NMR (400MHz, DMSO-d6) δ10.34(s,1H),8.96(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H), 2.58(s,3H),2.21–2.12(m,2H),1.81–1.53(m,3H),1.46–1.37(m,1H),1.34(s,3H).

[0337] Step 2: Synthesis of tert-butyl 6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoate

[0338] Intermediate compound 19-1 (100 mg, 309.25 μmol) and O-tert-butyl-N,N'-diisopropylisourea (619.48 mg, 3.09 mmol) were dissolved in a mixture of dichloromethane (5 mL) and tert-butanol (0.5 mL). The reaction mixture was stirred at room temperature overnight. LC-MS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 1 / 9 to 4 / 5) to afford compound 019 (35 mg).

[0339] MS (ESI): m / z = 380.10 [M+H] + .

[0340] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),8.96(d,J=2.1Hz,1H),8.41(d,J=2.1Hz,1H),5.85(s,1H),2.58( s,3H),2.21–2.12(m,2H),1.80–1.69(m,1H),1.69–1.61(m,1H),1.61–1.51(m,1H),1.43–1.32(m,13H)

[0341] Example 20: Synthesis of ethyl 6-[(6-cyano-5-(methoxy)pyridin-3-yl)amino]-5-hydroxy-5-methyl-6-oxohexanoate (Compound 020)

[0342] Step 1: Synthesis of ethyl 6-[(6-cyano-5-(methoxy)pyridin-3-yl)amino]-5,6-dioxohexanoate

[0343] Intermediate compound 20-1 (946.25 mg, 6.34 mmol) was added to tetrahydrofuran (20 mL) and precooled at 0°C for 15 min. Thionyl chloride (1.20 g, 10.06 mmol) was added and stirred at 0°C for 2 h. Triethylamine (1.36 g, 13.41 mmol) was added and stirred on ice for 10 min. Intermediate compound 5-5 (500 mg, 2.65 mmol) was added and stirred at room temperature for 12 h. LCMS indicated complete conversion. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The organic phases were combined and concentrated, and the residue was purified by column chromatography (silica, petroleum ether / ethyl acetate = 1 / 1) to afford the title compound 20-2 (940 mg).

[0344] MS (ESI): m / z = 320.1 [M+H] + .

[0345] 1 H NMR (400MHz, DMSO-d6) δ11.07(s,1H),8.76(d,J=1.9Hz,1H),8.24(d,J=1.9Hz,1H),4.08–4.01(m,2 H),3.94(s,3H),2.97(t,J=7.2Hz,2H),2.37(t,J=7.4Hz,2H),1.86–1.76(m,2H),1.19–1.17(m,3H).

[0346] Step 2: Synthesis of 6-[(6-cyano-5-(methyloxy)pyridin-3-yl)amino]-5-hydroxy-5-methyl-6-oxo-hexanoic acid ethyl ester

[0347] Intermediate compound 20-2 (200 mg, 626.35 μmol) was added to tetrahydrofuran (8 mL). Methylmagnesium bromide (3 M in Et2O, 627 μL) was added at -50°C and the reaction was stirred at -40°C for 12 h. LCMS indicated complete conversion. The reaction was quenched with saturated ammonium chloride solution, diluted with water (20 mL), and extracted with ethyl acetate (20 mL x 3). The organic phases were combined and concentrated, and the residue was purified by column chromatography (silica, petroleum ether / ethyl acetate = 3 / 1) to afford the target compound 020 (90 mg).

[0348] MS (ESI): m / z = 336.1 [M+H] + .

[0349] 1 H NMR (400MHz, DMSO-d6) δ10.29(s,1H),8.75(d,J=1.9Hz,1H),8.29(d,J=1.9Hz,1H),5.85(s,1H),4.07–3.99( m,2H),3.93(s,3H),2.33–2.21(m,2H),1.82–1.63(m,2H),1.63–1.40(m,2H),1.35(s,3H),1.20–1.10(m,3H).

[0350] Example 21: Synthesis of 5-[[6-cyano-5-(trifluoromethyl)pyridin-3-yl]carbamoyl]-6,6,6-trifluoro-5-hydroxyhexanoic acid ethyl ester (Compound 021)

[0351] Intermediate compound 5-6 (50 mg, 139.94 μmol) and (trifluoromethyl)trimethylsilane (59 mg, 419.83 μmol) were added to tetrahydrofuran (0.5 mL). Cesium fluoride (5 mg, 32.91 μmol) was added under nitrogen atmosphere, and the reaction solution was stirred at 25°C for 3 h. LCMS showed that the reaction was complete. Water (0.5 mL) was added to dilute the reaction solution, the reaction solution was filtered, and the filtrate was concentrated. The residue was purified by Prep-HPLC [Phenomenex C 18 , 5 μm silica, 30 mm diameter, 80 mm length; using a mixture of water (containing 7 mmol / L NH4HCO3) and acetonitrile (acetonitrile content: 60%-70%) as eluent, elution time: 8 minutes] to purify the target compound 021 (5 mg).

[0352] MS (ESI): m / z = 428.00 [M+H] + .

[0353] 1 H NMR(400MHz,Chloroform-d)δ9.45(s,1H),9.00–8.93(m,1H),8.83–8.75(m,1H),6.16(s,1H), 4.36–4.08(m,2H),2.59–2.38(m,3H),2.01–1.89(m,1H),1.71–1.58(m,2H),1.39–1.24(m,3H).

[0354] Example 22: N 1 -(6-cyano-5-(trifluoromethyl)pyridin-3-yl)-2-hydroxy-N 6 Synthesis of 2-dimethyladipamide (Compound 022)

[0355] Step 1: Synthesis of 6-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-5-hydroxy-5-methyl-6-oxohexanoic acid

[0356] Compound 5-7 (100 mg, 267.86 μmol) was dispersed in a mixture of tetrahydrofuran (3 mL) and water (1 mL). Lithium hydroxide monohydrate (12.84 mg, 306.01 μmol) was added and stirred at room temperature overnight. LC-MS indicated the reaction was complete. The reaction solution was poured into 0.1 M aqueous hydrogen chloride solution (15 mL) and extracted with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield compound 22-1 (80 mg).

[0357] MS (ESI): m / z = 346.10 [M+H] + .

[0358] 1 H NMR (400MHz, DMSO-d6) δ9.40(d,J=2.2Hz,1H),8.91(d,J=2.3Hz,1H),2.22–2.13(m,2H),1.81–1.54(m,3H),1.48–1.36(m,1H),1.36(s,3H).

[0359] Step 2: N 1 -(6-cyano-5-(trifluoromethyl)pyridin-3-yl)-2-hydroxy-N 6 Synthesis of 2-dimethyladipamide

[0360] Intermediate compound 22-1 (50 mg, 144.81 μmol) was dissolved in tetrahydrofuran (4 mL), cooled to 0°C, and thionyl chloride (18 mg, 151.31 μmol) was added dropwise. The mixture was allowed to react at room temperature for 6 h. Methylamine solution (1 M in THF, 1.5 mL) was added to the reaction mixture, and the mixture was stirred at room temperature for 3 h. LC-MS indicated the reaction was complete. The reaction mixture was poured into ice-cold 1 M hydrochloric acid solution (30 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The residue was purified by Prep-HPLC using a YMC-TAR column (5 μm silica, 30 mm diameter, 150 mm length; eluent: a decreasingly polar mixture of water (containing 7 mmol / L NH₄HCO₃) and acetonitrile; acetonitrile gradient from 30% to 60% over 7 minutes) to afford the target compound 022 (15 mg).

[0361] MS (ESI): m / z = 359.10 [M+H] + .

[0362] 1 H NMR (400MHz, DMSO-d6) δ9.39 (s, 1H), 8.91 (d, J = 2.3Hz, 1H), 7.69 (s, 1H), 5.90 (s, 1H),2.55-2.51(m,3H),2.10-2.01(m,2H),1.74-1.50(m,3H),1.42-1.32(m,4H).

[0363] Example 23: N 1 Synthesis of -(6-cyano-5-(trifluoromethyl)pyridin-3-yl)-2-hydroxy-2-methyladipamide (Compound 023)

[0364] Compound 22-1 (50 mg, 144.81 μmol) was dissolved in tetrahydrofuran (4 mL), cooled to 0°C, and thionyl chloride (18 mg, 151.31 μmol) was added dropwise. The mixture was allowed to react at room temperature for 6 h. Aqueous ammonia (2 mL, 35%) was added to the reaction mixture, and the mixture was stirred at room temperature for 3 h. LC-MS indicated the reaction was complete. The reaction mixture was poured into ice-cold 1 M hydrochloric acid (30 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The residue was purified by Prep-HPLC using a YMC-TAR column (5 μm silica, 30 mm diameter, 150 mm length; eluent: a decreasingly polar mixture of water (containing 7 mmol / L NH₄HCO₃) and acetonitrile; acetonitrile gradient from 40% to 70% over 12 minutes) to afford the target compound 023 (15 mg).

[0365] MS (ESI): m / z = 345.10 [M+H] + .

[0366] 1 H NMR (400MHz, DMSO-d6) δ10.70(s,1H),9.40(d,J=2.3Hz,1H),8.91(d,J=2.3Hz,1H),7.23( s,1H),6.69(s,1H),5.92(s,1H),2.06-1.97(m,2H),1.79-1.52(m,3H),1.43-1.33(m,4H).

[0367] Example 24: 6-[(6-cyano-5-methylselenopyridin-3-yl)amino]-5-hydroxy-5-methyl-6-oxohexanoic acid ethyl ester (Compound 024)

[0368] Step 1: Synthesis of 5-amino-3-fluoropyridine-2-carbonitrile

[0369] 6-Bromo-5-fluoropyridin-3-amine (5 g, 26.18 mmol), zinc cyanide (6.15 g, 52.36 mmol), zinc powder (166.47 mg, 2.54 mmol), tris(dibenzylideneacetone)dipalladium (1.2 g, 1.31 mmol), and 1,1'-bis(diphenylphosphino)ferrocene (1.45 g, 2.62 mmol) were added to N,N-dimethylformamide (50 mL). The reaction mixture was allowed to react at 100°C under an argon atmosphere for 16 hours. LCMS indicated the reaction was complete. The reaction mixture was poured into water (500 mL) and extracted with ethyl acetate (200 mL x 4). The combined organic phases were washed with saturated brine (400 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The crude product was purified by column chromatography (silica, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to give the title compound 24-2 (2.9 g).

[0370] MS (ESI): m / z = 138.10 [M+H] + .

[0371] Step 2: Synthesis of 5-amino-3-(methylseleno)pyridine-2-carbonitrile

[0372] 5-Amino-3-fluoropyridine-2-carbonitrile (1.6 g, 11.67 mmol), dimethyl diselenide (6.68 g, 35.53 mmol), N,N-diisopropylethylamine (4.52 g, 35.01 mmol), and potassium carbonate (4.83 g, 35.01 mmol) were added to N,N-dimethylformamide (20 mL). The reaction mixture was stirred at 100°C for 18 hours. LCMS indicated the reaction was complete. The reaction mixture was poured into water and extracted with ethyl acetate (200 mL x 4). The combined organic phases were washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The crude product was purified by column chromatography (silica, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to afford the title compound 24-3 (2 g).

[0373] MS (ESI): m / z = 214.10 [M+H] + .

[0374] 1 H NMR (400MHz, DMSO-d6) δ7.79(d,J=2.3Hz,1H),6.95(d,J=2.4Hz,1H),6.49(s,2H),2.39(s,3H).

[0375] Step 3: Synthesis of ethyl 6-[(6-cyano-5-methylselenopyridin-3-yl)amino]-5,6-dioxohexanoate

[0376] 6-Ethoxy-2,6-dioxohexanoic acid (166.34 mg, 883.94 μmol) and 5-amino-3-(methylseleno)pyridine-2-carbonitrile (100 mg, 471.45 μmol) were dissolved in N,N-dimethylacetamide (3 mL). Thionyl chloride (112.18 mg, 942.91 μmol) was added dropwise at -10°C. The reaction mixture was stirred at -10°C for 18 hours. LCMS indicated the reaction was complete. The reaction mixture was poured into water (50 mL) and extracted with ethyl acetate (30 mL x 4). The combined organic phases were washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to afford the title compound 24-4 (220 mg).

[0377] MS (ESI): m / z = 384.10 [M+H] + .

[0378] Step 4: Synthesis of ethyl 6-[(6-cyano-5-methylselenopyridin-3-yl)amino]-5-hydroxy-5-methyl-6-oxohexanoate

[0379] Intermediate 24-4 (200 mg, 523.18 μmol) was added to tetrahydrofuran (5 mL), and methylmagnesium bromide in tetrahydrofuran (3 M, 524 μL) was added dropwise at -78°C. The reaction was stirred at -78°C for 18 hours. LCMS indicated the reaction was complete. The reaction was diluted with ethyl acetate (10 mL), quenched by the addition of saturated ammonium chloride solution (10 mL) at low temperature, and extracted with ethyl acetate (20 mL x 4). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The crude product was purified by prep-HPLC (Phenomenex C18, 5 μm silica, 30 mm diameter, 80 mm length; using a mixture of water (containing 0.05% ammonium bicarbonate) and acetonitrile (acetonitrile content: 38-68%) as eluent) to afford the target compound 024 (60 mg).

[0380] MS (ESI): m / z = 400.10 [M+H] + .

[0381] 1 H NMR(400MHz,Chloroform-d)δ9.15(s,1H),8.59(d,J=2.3Hz,1H),8.40(d,J=2.3Hz,1H),4.18–4.12(m,2H),3.55( s,1H),2.48(s,3H),2.42–2.35(m,2H),2.10–2.01(m,1H),1.82–1.62(m,3H),1.51(s,3H),1.26(t,J=7.2Hz,3H).

[0382] Example 25: (5R)-6-[(6-cyano-5-methylsulfonylpyridin-3-yl)amino]-5-hydroxy-5-methyl-6-oxo-hexanoic acid ethyl ester or (5S)-6-[(6-cyano-5-methylsulfonylpyridin-3-yl)amino]-5-hydroxy-5-methyl-6-oxo-hexanoic acid ethyl ester (Compound 025)

[0383] Compound 014 (90.00 mg, 256.10 μmol) was added to dichloromethane (1 mL). m-Chloroperbenzoic acid (155.99 mg, 768.31 μmol, 85% content) was added under ice-cooling. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was filtered, and the filtrate was washed with water (1 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by column chromatography (silica, petroleum ether / ethyl acetate = 2 / 1) to obtain compound 025 (59 mg).

[0384] MS (ESI): m / z = 384.2 [M+H]+ .

[0385] 1 H NMR (400MHz, DMSO-d6) δ10.74(s,1H),9.39–9.30(m,1H),9.13–9.05(m,1H),5.89(s,1H),4.10–3.95(m,2H),3.44( s,3H),2.31–2.23(m,2H),1.81–1.66(m,2H),1.64–1.54(m,1H),1.50–1.39(m,1H),1.36(s,3H),1.20–1.11(m,3H).

[0386] Example 26: (5R)-6-[(6-cyano-5-methylsulfinylpyridin-3-yl)amino]-5-hydroxy-5-methyl-6-oxo-hexanoic acid ethyl ester or (5S)-6-[(6-cyano-5-methylsulfinylpyridin-3-yl)amino]-5-hydroxy-5-methyl-6-oxo-hexanoic acid ethyl ester (Compound 026)

[0387] Compound 014 (50 mg, 142.28 μmol) was added to dichloromethane (0.5 mL). m-Chloroperbenzoic acid (23.11 mg, 113.82 μmol, 85% content) was added under ice-cooling. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was filtered, and the filtrate was washed with water (1 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The mixture was purified by column chromatography (silica, petroleum ether / ethyl acetate = 2 / 1) to obtain compound 026 (5 mg).

[0388] MS (ESI): m / z = 368.1 [M+H] + .

[0389] 1 H NMR (400MHz, DMSO-d6) δ10.64(s,1H),9.21–9.11(m,1H),9.03–8.90(m,1H),5.85(s,1H),4.14–3.91(m,2H),2.91( s,3H),2.32–2.23(m,2H),1.84–1.65(m,2H),1.65–1.54(m,1H),1.50–1.41(m,1H),1.36(s,3H),1.20–1.11(m,3H).

[0390] Example 27: Ethyl 7-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-6-hydroxy-6-methyl-7-oxoheptanoate (Compound 027)

[0391] Step 1: Synthesis of triethyl 1-oxopentane-1,2,5-tricarboxylate

[0392] Starting material 27-1 (2 g, 9.89 mmol) was added to tetrahydrofuran (10 mL). Sodium ethoxide (672.45 mg, 9.89 mmol) was added under ice-water bath. The mixture was stirred at room temperature for 30 minutes, and diethyl oxalate (1.45 g, 9.89 mmol) was added. The reaction mixture was stirred at room temperature for 18 hours. 20 mL of water was added to dilute the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic layers were combined and washed with water (30 mL x 2), then with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound 27-2 (2.99 g).

[0393] 1 H NMR(400MHz,Chloroform-d)δ4.45–4.29(m,2H),4.25–4.09(m,4H),4.02(t,J=7.0 Hz,1H),2.42–2.27(m,2H),2.04–1.85(m,2H),1.79–1.59(m,2H),1.44–1.17(m,9H)

[0394] Step 2: Synthesis of 2-oxopimelic acid

[0395] Intermediate 27-2 (6 g, 19.85 mmol) was added to 4 M aqueous HCl (60 mL). The reaction mixture was stirred at 70°C for 18 hours. LCMS indicated the reaction was complete. Dilute with 20 mL of water and extract with ethyl acetate (20 mL x 3). The combined organic layers were washed with water (30 mL x 2), saturated brine (50 mL), and dried over anhydrous sodium sulfate. Filter and concentrate under reduced pressure to obtain the title compound 27-3 (3 g).

[0396] MS (ESI): m / z = 173.00 [MH] - .

[0397] Step 3: Synthesis of diethyl 2-oxopimelate

[0398] Intermediate 27-3 (2.8 g, 16.08 mmol) and concentrated sulfuric acid (3.15 g, 32.16 mmol, 1.71 mL) were added to ethanol (25 mL). The reaction mixture was stirred at 50°C for 18 hours. LCMS showed that the reaction was complete. The reaction mixture was cooled and poured into water (20 mL). It was extracted with ethyl acetate (30 mL*3). The organic layers were combined and washed with water (40 mL*2), washed with saturated brine (40 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, concentrated under reduced pressure, and purified by column chromatography (silica, ethyl acetate / petroleum ether = 3 / 1) to obtain the title compound 27-4 (1.6 g).

[0399] MS (ESI): m / z = 231.1 [M+H] + .

[0400] 1 H NMR (400MHz, Chloroform-d) δ4.37–4.22(m,2H),4.18–4.08(m,2H),2.92–2.81(m,2H),2.37–2.24(m,2H),1.75–1.55(m,4H),1.44–1.19(m,6H).

[0401] Step 4: Synthesis of 7-ethoxy-2,7-dioxoheptanoic acid

[0402] Intermediate 27-4 (166 mg, 720.93 μmol) was added to ethanol (2 mL) and water (1 mL). Potassium hydroxide (40.45 mg, 720.93 μmol) was added at 0°C, and the reaction mixture was stirred at room temperature for 1 hour. LCMS indicated the reaction was complete. The reaction mixture was washed once with ethyl acetate (20 mL), and the aqueous phase was acidified (pH = 2-3) with dilute hydrochloric acid (10 mL). The mixture was then extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the title compound 27-5 (130 mg).

[0403] MS (ESI): m / z = 201.20 [MH] - .

[0404] 1 H NMR (400MHz, Chloroform-d) δ4.27–4.07(m,2H),3.01–2.92(m,2H),2.39–2.31(m,2H),1.78–1.59(m,2H),1.35–1.18(m,3H).

[0405] Step 5: Synthesis of ethyl 7-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-6,7-dioxoheptanoate

[0406] Intermediate 27-5 (324.18 mg, 1.60 mmol) and 5-amino-3-(trifluoromethyl)pyridine-2-carbonitrile (200 mg, 1.07 mmol) were added to N,N-dimethylacetamide (3 mL). Thionyl chloride (216.17 mg, 1.82 mmol) was added under ice-water bath. The reaction mixture was stirred under ice-water bath for 18 hours. LCMS indicated the reaction was complete. The reaction mixture was poured into ice water and extracted with ethyl acetate (5 mL*3). The organic layers were combined and washed with water (40 mL*2), then with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the title compound 27-6 (400 mg).

[0407] MS (ESI): m / z = 372.10 [M+H] + .

[0408] Step 6: Synthesis of ethyl 7-((6-cyano-5-(trifluoromethyl)pyridin-3-yl)amino)-6-hydroxy-6-methyl-7-oxoheptanoate

[0409] Intermediate 27-6 (200 mg, 538.63 μmol) was added to tetrahydrofuran (3 mL). Methylmagnesium bromide tetrahydrofuran solution (3 M, 540 μL) was added to the reaction solution at -78°C. The reaction solution was stirred at -78°C for 18 hours. LCMS showed that the reaction was complete. 30 mL of saturated aqueous ammonium chloride solution was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL*3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica, ethyl acetate / petroleum ether = 5 / 1) to obtain the title compound 027 (40 mg).

[0410] MS (ESI): m / z = 388.10 [M+H] + .

[0411] 1 H NMR (400MHz, DMSO-d6) δ10.70(s,1H),9.39(d,J=2.2Hz,1H),8.91(d,J=2.3Hz,1H),5.87(s,1H),4.01(q ,J=7.1Hz,2H),2.30–2.22(m,2H),1.82–1.70(m,1H),1.68–1.41(m,3H),1.36(s,3H),1.29–1.06(m,5H).

[0412] Example 28: 5-[(6-cyano-5-methylthiopyridin-3-yl)carbamoyl]-6,6,6-trifluoro-5-hydroxyhexanoic acid ethyl ester (Compound 028)

[0413] Intermediate 13-5 (1 g, 2.98 mmol) and (trifluoromethyl)trimethylsilane (2.12 g, 14.91 mmol) were added to tetrahydrofuran (20 mL). Cesium fluoride (9.85 mg, 64.84 μmol) was added under a nitrogen atmosphere, and the reaction solution was stirred at 25°C for 3 hours. LCMS showed that the reaction was complete. The reaction solution was poured into ethyl acetate (20 mL) and water (20 mL). The layers were separated, and the aqueous phase was extracted twice with ethyl acetate (20 mL). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (silica, petroleum ether / ethyl acetate = 3 / 2) to obtain compound 028 (120 mg).

[0414] MS (ESI): m / z = 406.1 [M+H] + .

[0415] 1 H NMR(400MHz,DMSO-d6)δ10.66(s,1H),9.01–8.89(m,1H),8.48–8.31(m,1H),7.65(s,1H),4.09–3.94(m,2H), 2.60(s,3H),2.37–2.29(m,2H),2.20–2.06(m,1H),1.89–1.68(m,2H),1.47–1.34(m,1H),1.20–1.12(m,3H).

[0416] Example 29: 6-(4-cyano-3-methylthio-anilino)-5-hydroxy-5-methyl-6-oxohexanoic acid ethyl ester (Compound 029)

[0417] Step 1: Synthesis of 6-(4-cyano-3-methylthio-anilino)-5,6-dioxo-hexanoic acid ethyl ester

[0418] Intermediate 5-5 (1.01 g, 5.36 mmol) was added to N,N-dimethylacetamide (5 mL). Thionyl chloride (724.42 mg, 6.09 mmol) was added at -10°C, followed by 4-amino-2-methylthio-benzonitrile (29-1, 500 mg, 3.04 mmol). The reaction mixture was stirred at -78°C for 15 hours. LCMS indicated the reaction was complete. Ethyl acetate (50 mL) was added for dilution, quenched with saturated sodium bicarbonate solution (50 mL), and extracted with ethyl acetate (50 mL x 4). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (silica, petroleum ether / ethyl acetate = 1 / 1) to afford Intermediate 29-2 (600 mg).

[0419] MS (ESI): m / z = 335.20 [M+H] + .

[0420] 1 H NMR (400MHz, DMSO-d6) δ10.84(s,1H),7.96(d,J=1.9Hz,1H),7.84(dd,J=8.6,1.9Hz,1H),7.77(d,J=8.5Hz,1H),4.06 (q,J=7.1Hz,2H),2.97(t,J=7.2Hz,2H),2.56(s,3H),2.36(t,J=7.4Hz,2H),1.85–1.75(m,2H),1.18(t,J=7.1Hz,3H).

[0421] Step 2: Synthesis of ethyl 6-(4-cyano-3-methylthio-anilino)-5-hydroxy-5-methyl-6-oxohexanoate

[0422] Intermediate 29-2 (400 mg, 1.20 mmol) was added to tetrahydrofuran (4 mL), cooled to -78°C, and methylmagnesium bromide in diethyl ether (3 M, 1.2 mL) was added under constant temperature. The reaction mixture was stirred at -78°C for 15 hours. LCMS indicated completion of the reaction. Ethyl acetate (50 mL) was added for dilution, and the mixture was quenched by the addition of saturated ammonium chloride solution (50 mL) at low temperature. The mixture was then extracted with ethyl acetate (50 mL x 4). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC (Phenomenex C18, 5 μm silica, 30 mm diameter, 80 mm length; eluent: a mixture of water (containing 0.05% ammonium bicarbonate) and acetonitrile (50-80% acetonitrile)) to afford the target compound 029 (110 mg).

[0423] MS (ESI): m / z = 351.20 [M+H]+ .

[0424] 1 H NMR (400MHz, DMSO-d6) δ10.02(s,1H),7.88(d,J=1.9Hz,1H),7.79(dd,J=8.6,1.9Hz,1H),7.65(d,J=8.5Hz,1H),5.76(s,1H),3.97(q,J=7. 1Hz,2H),2.50(s,3H),2.21(t,J=7.1Hz,2H),1.73–1.58(m,2H),1.55–1.47(m,1H),1.41–1.31(m,1H),1.28(s,3H),1.10(t,J=7.1Hz,3H).

[0425] Example 30: 6-(4-cyano-3-methoxy-anilino)-5-hydroxy-5-methyl-6-oxohexanoic acid ethyl ester (Compound 030)

[0426] Step 1: Synthesis of 6-(4-cyano-3-methoxy-anilino)-5,6-dioxo-hexanoic acid ethyl ester

[0427] 6-Ethoxy-2,6-dioxohexanoic acid (971.24 mg, 5.16 mmol) was added to N,N-dimethylacetamide (5 mL), and the reaction mixture was cooled to -10°C. Thionyl chloride (802.97 mg, 6.75 mmol) was added dropwise, followed by 4-amino-2-methoxybenzonitrile (500 mg, 3.37 mmol). The reaction mixture was stirred at -78°C for 15 hours. LCMS indicated the reaction was complete. The mixture was quenched with saturated sodium bicarbonate solution (50 mL) and extracted with ethyl acetate (50 mL x 4). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (silica, petroleum ether / ethyl acetate = 1 / 1) to afford Intermediate 30-2 (600 mg).

[0428] MS (ESI): m / z = 319.20 [M+H] + .

[0429] 1H NMR (400MHz, DMSO-d6) δ10.77(s,1H),7.79(d,J=1.8Hz,1H),7.69(d,J=8.5Hz,1H),7.59(dd,J=8.6,1.8Hz,1H),4.05 (q,J=7.1Hz,2H),3.87(s,3H),2.95(t,J=7.2Hz,2H),2.35(t,J=7.4Hz,2H),1.84–1.73(m,2H),1.17(t,J=7.1Hz,3H).

[0430] Step 2: Synthesis of ethyl 6-(4-cyano-3-methoxy-anilino)-5-hydroxy-5-methyl-6-oxohexanoate

[0431] Intermediate 30-2 (500 mg, 1.57 mmol) was added to tetrahydrofuran (5 mL). Methylmagnesium bromide in tetrahydrofuran (3 M, 1.57 mL) was added at -78°C. The reaction was stirred at -78°C for 15 hours. LCMS indicated the reaction was complete. Ethyl acetate (50 mL) was added for dilution, and the mixture was quenched by saturated ammonium chloride solution (50 mL) at low temperature. The mixture was then extracted with ethyl acetate (50 mL x 4). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC (Phenomenex C18, 5 μm silica, 30 mm diameter, 80 mm length; eluent: a mixture of water (containing 0.05% ammonium bicarbonate) and acetonitrile (50-80% acetonitrile)) to afford the target compound 030 (110 mg).

[0432] MS (ESI): m / z = 335.20 [M+H] + .

[0433] 1 H NMR(400MHz, DMSO-d6)δ9.99(s,1H),7.80(d,J=1.7Hz,1H),7.66–7.56(m,2H),5.81(s,1H),4.02(q,J=7.1Hz,2H ),3.86(s,3H),2.26(t,J=7.1Hz,2H),1.79–1.65(m,2H),1.61–1.38(m,2H),1.33(s,3H),1.15(t,J=7.1Hz,3H).

[0434] Example 31: Ethyl 6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-6-oxohexanoate (Compound 031)

[0435] Compound 13-5 (10 mg, 29.82 μmol) was added to methanol (1 mL). Sodium borohydride (2.26 mg, 59.63 μmol) was added under ice-water bath. The reaction solution was stirred at room temperature for 1 hour. LCMS showed that the reaction was complete. 10 mL of water was added to the reaction solution under ice-water bath to quench the reaction solution, and the solution was extracted with ethyl acetate (200 mL*3). The organic phases were combined, washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound 031 (9 mg).

[0436] MS (ESI): m / z = 338.20 [M+H] + .

[0437] Example 32: Isopropyl 6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-6-oxohexanoate (Compound 032)

[0438] Step 1: Synthesis of 6-(6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-6-oxohexanoic acid

[0439] Compound 031 (50 mg, 148.19 μmol) and lithium hydroxide monohydrate (7.11 mg, 169.45 μmol) were added to tetrahydrofuran (1 mL) and deionized water (1 mL). The reaction solution was stirred at room temperature for 2 hours. LCMS showed that the reaction was complete. 10 mL of water was added to dilute, and the mixture was extracted with ethyl acetate (50 mL). 1 M HCl aqueous solution (10 mL) was added to the aqueous phase to adjust the system to acidic (pH = 2-3), and extracted with ethyl acetate (20 mL * 3). The organic phases were combined, washed with saturated brine (20 mL * 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound 32-1 (35 mg).

[0440] MS (ESI): m / z = 309.6 [M+H] + .

[0441] 1 H NMR(400MHz,DMSO-d6)δ8.89(d,J=2.1Hz,1H),8.36(d,J=2.1Hz,1H),4.11–4.0 3(m,1H),2.57(s,3H),2.14–2.05(m,2H),1.78–1.71(m,1H),1.65–1.55(m,3H).

[0442] Step 2: Synthesis of isopropyl 6-((6-cyano-5-(methylthio)pyridin-3-yl)amino)-5-hydroxy-6-oxohexanoate

[0443] Intermediate 32-1 (50 mg, 161.63 μmol) and concentrated sulfuric acid (15.85 mg, 161.63 μmol) were added to isopropanol (1 mL). The reaction was stirred at room temperature for 6 hours. LCMS indicated the reaction was complete. The reaction solution was filtered, and the filtrate was purified by Prep-HPLC [Phenomenex C18, 5 μm silica, 30 mm diameter, 80 mm length; using a mixture of pure water and acetonitrile (acetonitrile content: 60%-70%) as the eluent] to obtain the target compound 032 (10 mg).

[0444] MS (ESI): m / z = 351.7 [M+H] + .

[0445] 1 H NMR(400MHz, DMSO-d6)δ8.89(d,J=2.1Hz,1H),8.35(d,J=2.1Hz,1H),6.03(s,1H),4.93–4.82(m,1H),4.1 3–4.05(m,1H),2.58(s,3H),2.33–2.22(m,2H),1.79–1.68(m,1H),1.70–1.53(m,3H),1.20–1.14(m,6H).

[0446] Experimental Example 1: Effects of the Disclosed Compounds on AR Antagonistic Activity

[0447] (1) Materials and reagents:

[0448] (2) Instruments:

[0449] (3) Experimental methods:

[0450] Day 1: Cell plating

[0451] 8x10 5 LNCaP cells.

[0452] Day 2: Transfection

[0453] Using Lipo 3000 and P3000 (Lipofectamine TM 3000) transfection reagent and AR and PSA ARE1 reporter gene plasmids (Bio (Anhui) Co., Ltd.) (a total of 2.5 μg of plasmids and 1.25 μg of each plasmid were added to each well) were incubated at room temperature for 10 minutes, and then 250 μL of the above transfection reagent was added dropwise to each well containing cells.

[0454] Day 3: Cell treatment, plating and drug addition

[0455] The cells were washed with PBS and the medium was replaced with RPMI-1640 without phenol red and 5% charcoal-stripped serum. 4 cells / well), and the cells were re-plated into a 96-well plate (Corning, 3603).

[0456] After plating the cells, add the test compound (or DMSO or positive control) and incubate for at least 30 minutes (incubator, 37°C, 5% CO2). Then, add DHT (dihydrotestosterone) to a final concentration of 1 nM and continue incubating in a CO2 incubator. The final plate volume is 100 μL.

[0457] The test compound was prepared as a 10 mM stock solution with a final concentration of 3.3 μM or 6.7 μM. The negative control was 0.1% DMSO. The positive control was 30 μM Enzalutamide.

[0458] Day 4: Luciferase signal detection and data processing

[0459] After incubating the test compound (or DMSO or positive control) with DHT for 6 or 24 hours, the luciferase signal was detected using Vazyme Bright-Lite substrate. 70 μL of culture medium was added to each well, and the cells were shaken for 2 minutes before reading the luciferase signal.

[0460] (4) Data processing:

[0461] Check assay robustness using DMSO and low concentration control data:

[0462] Ave_H=Ave(DMSO)

[0463] Ave_L = Ave (30000 nM enzalutamide)

[0464] Inhibition rate (%) = (Ave_H-Sample) / (Ave_H-Ave_L) x 100%

[0465] Ave_H: measured value of negative control (0.1% DMSO);

[0466] Ave_L: measured value of the positive control (30 μM enzalutamide);

[0467] Sample: the measured value of the compound to be tested;

[0468] The activity test results of the test compounds are shown in Table 1.

[0469] Table 1 Activity test results of the compounds disclosed herein Note: a represents incubation time: 24h, test concentration: 3.3μM; b represents incubation time: 6h, test concentration: 6.7μM; c represents incubation time: 24h, test concentration: 6.7μM.

[0470] From the above results, it can be seen that the compounds disclosed herein have AR antagonistic activity.

[0471] Experimental Example 2: Metabolic Stability Determination of the Compounds of the Present Disclosure in Liver Microsomes

[0472] The metabolic stability of the disclosed compounds in liver microsomes was determined using the following test method.

[0473] 1. Test materials and instruments

[0474] 1. Source of liver microsomes: human liver microsomes (Corning 452117)

[0475] 2.Na2HPO4 (Tianjin Guangfu Fine Chemical Research Institute 20180130)

[0476] 3. KH2PO4 (Tianjin Guangfu Fine Chemical Research Institute 20180920)

[0477] 4.MgCl2 (Tianjin Guangfu Fine Chemical Research Institute 20191216)

[0478] 5. NADPH (Solarbio 1216C022)

[0479] 6. Positive control compound: Verapamil (Sigma MKBV4993V)

[0480] 7. AB Sciex Triple Quad 4000 LC / MS

[0481] 2. Test steps

[0482] 1. Preparation of 100 mM phosphate buffer (PBS): Weigh 7.098 g of Na₂HPO₄ and dissolve it in 500 mL of pure water by sonication to prepare Solution A. Weigh 3.400 g of KH₂PO₄ and dissolve it in 250 mL of pure water by sonication to prepare Solution B. Place Solution A on a stirrer and slowly add Solution B until the pH reaches 7.4 to prepare 100 mM PBS buffer.

[0483] 2. Preparation of reaction system

[0484] Prepare the reaction system according to the table below

[0485] 3. Preincubate the reaction system in a 37°C water bath for 10 minutes. Add 40 μL of 10 mM NADPH solution (dissolved in 100 mM phosphate buffer) to the reaction system for a final NADPH concentration of 1 mM. Replace the NADPH solution with 40 μL of phosphate buffer as a negative control. This negative control is used to eliminate the effects of the chemical stability of the compound itself.

[0486] 4. 4 μL of 100 μM compounds of the present disclosure and the positive control compound verapamil were added to the reaction system to initiate the reaction. The final concentration of the compounds was 1 μM.

[0487] 5. After thorough mixing on a vortex shaker at 0.5, 15, 30, 45, and 60 minutes, remove 50 μL of the incubation sample and terminate the reaction with 4x volume of glacial acetonitrile containing the internal standard. Centrifuge the sample at 3,220 g for 45 minutes. After centrifugation, transfer 90 μL of the supernatant to the injection plate, add 90 μL of ultrapure water, and mix thoroughly before LC-MS / MS analysis.

[0488] All data were calculated using Microsoft Excel software. Peak areas were detected by extracting ion spectra. The in vitro half-life (T) of the test compound was determined by linear fitting the natural logarithm of the elimination percentage of the test compound with time. 1 / 2 ).

[0489] In vitro half-life 1 / 2 ) is calculated by the slope k: in vitro T 1 / 2 =0.693 / k

[0490] In vitro intrinsic clearance (CL int , unit: μL / min / mg protein) was calculated using the following formula: int =k×incubation volume / enzyme protein content;

[0491] T calculated by the above formula 1 / 2 and CL int See Table 2 for values.

[0492] Table 2 Half-life and intrinsic clearance values ​​of the disclosed compounds in liver microsomes

[0493] From the above results, it can be seen that the disclosed compounds have the characteristics of high clearance rate and rapid metabolism, which avoids the accumulation of drugs in the body due to long-term administration, thereby affecting the systemic androgen receptor signaling pathway, thereby improving the safety of the drug.

[0494] Experimental Example 3 Whole Blood Stability

[0495] Test steps:

[0496] 1. Preparation of Stock Solutions

[0497] 1 mM stock solutions of test compounds were prepared in DMSO, and 1 mM stock solutions of the control compound propantheline were prepared in acetonitrile.

[0498] 2. Stability determination method

[0499] 4 μL of test compound stock solution (or control compound stock solution) was added to 796 μL of pre-incubated whole blood (CD-1 mouse whole blood or human whole blood) to achieve a final concentration of 5 μM. The final concentration of the solvent was 0.5%. At various time points (including 5, 15, 30, 60, and 120 min), 50 μL aliquots of the sampled whole blood were added to new tubes and incubated in a 37°C water bath with shaking at 60 rpm. The experiment was repeated twice. At the designated time points, 300 μL of room temperature quenching solution (acetonitrile solution containing internal standards (100 nM alprazolam, 500 nM labetalol, and 2 μM ketoprofen)) was added to the sampled whole blood samples to terminate the reaction. 50 μL of the sampled whole blood was added to new tubes containing 300 μL of room temperature quenching solution, and the time 0 sample was prepared. All samples were vortexed for 5 minutes. The samples in the plate were centrifuged at 3220 g for 30 minutes at 4°C to precipitate proteins. 100 μL of the supernatant was transferred to a new 96-well plate containing 100 μL of water for LC-MS / MS analysis.

[0500] 3. Data Analysis

[0501] All calculations were performed using Microsoft Excel. Peak area ratios were determined from extracted ion chromatograms. The percentage of compound remaining at each time point was calculated using the following formula:

[0502] Remaining percentage tmin (%) = peak area ratio tmin / peak area ratio 0min×100%

[0503] Wherein, peak area ratio t min: the peak area ratio of the reference compound and the test compound at t minutes;

[0504] Peak area ratio 0min: the peak area ratio of the control compound and the test compound at zero time point.

[0505] The slope value was determined by linear regression of the natural logarithm of the remaining percentage of the test compound versus the incubation time curve.

[0506] The in vitro half-life (in vitro t 1 / 2 ): in vitro t 1 / 2 =0.693 / k

[0507] Where: k is the rate constant (k = -slope value)

[0508] The half-lives of the disclosed compounds in CD-1 mouse or human whole blood are shown in Table 3.

[0509] Table 3. Stability of the disclosed compounds in CD-1 mice and human whole blood

[0510] From the above results, it can be seen that the compounds of the present disclosure have a short half-life in CD-1 mice and human whole blood, and have the characteristics of high clearance rate and rapid metabolism in the blood, which avoids the accumulation of drugs in the body due to long-term administration, thereby affecting the systemic androgen receptor signaling pathway, thereby improving the safety of the drugs.

[0511] Experimental Example 4 In vivo efficacy study

[0512] 4.1 Experimental Reagents

[0513] Solvent: Propylene glycol / ethanol (30:70, v / v)

[0514] 4.2 Experimental methods

[0515] Animal information: C57BL / 6 male mice, 6 weeks old, weighing approximately 18-20 g, were purchased from Shanghai Lingchang Biotechnology Co., Ltd. The mice were housed in an SPF-grade environment with separate ventilation in each cage. All animals had free access to standard certified commercial laboratory diet and free drinking water.

[0516] Skin preparation: After the mice have been adapted for 1-2 weeks, use a shaver to remove the hair in a 2 cm*3 cm area on the back of the mice. Confirm that the hair of the mice is in the resting phase (the skin is pink) and the skin is not damaged.

[0517] Dosing: The mice were divided and dosed on the third day after shaving. The disclosed compound 010 was applied to the shaved area twice a day, morning and evening, at a concentration of 0.5 wt% for 32 days. The control group was treated with the control vehicle. Each group consisted of 12 mice.

[0518] 4.3 Observation of mouse status and hair growth scoring

[0519] During the experiment, the condition of mice in each group was observed to see if there were any rashes, cracks, or dandruff on the skin.

[0520] After administration of Compound 010, mice were weighed, hair growth was scored, and photographs were taken on days 10, 13, 16, 20, 23, 27, and 32;

[0521] The scoring criteria are as follows:

[0522] 0 points: no growth, the skin of the shaved area is pink;

[0523] 1 point: the skin in the shaved area is gray (less than 20% growth);

[0524] 2 points: The skin of the shaved area is black (greater than 20% and less than 40%);

[0525] 3 points: The skin of the shaved area is black with a little hair growth (more than 40% but less than 60%);

[0526] 4 points: The skin in the shaved area is black with some hair growth (more than 60% but less than 80%);

[0527] 5 points: The hair in the shaved area has basically grown back (80% to 100% growth).

[0528] 4.4 Experimental Results

[0529] The results of the effects of the disclosed compounds on hair growth in mice are shown in Table 4.

[0530] Table 4 Effect of the disclosed compounds on hair growth in mice *P<0.05, **P<0.01, vs. vehicle control group

[0531] 4.5 Experimental Conclusion

[0532] In mice whose hair growth is in the telogen phase, the compound 010 of the present disclosure at a concentration of 0.5 wt % at 20 μL / cm 2 , BID administration by skin application significantly promoted hair growth. Compound 010 significantly promoted hair growth starting at day 32 (P < 0.01). Compound 010 did not significantly affect mouse body weight at the doses tested, nor did it cause any skin rash, cracking, or dandruff. Other compounds of the present disclosure also significantly promoted hair growth.

[0533] Although specific embodiments of the present disclosure have been illustrated and described, those skilled in the art will appreciate that, where feasible, the technical features described in one embodiment may be applied to another embodiment, or combined with the technical features described in another embodiment. Therefore, those skilled in the art will be able to make various changes and modifications to the embodiments of the present disclosure without departing from the spirit and scope of the present disclosure.

Claims

1. A compound represented by formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, in, X is selected from CH or N; Y is selected from OH, COOH, -CONH2, -COOR 3 or-CONR 3’ R 3 ; R 1 Selected from OH or -O-C1-C6 alkyl; R 2 is selected from H, deuterium, C1-C6 alkyl, C1-C6 deuterated alkyl or C1-C6 haloalkyl; R 3 is selected from C1-C6 alkyl, C1-C6 deuterated alkyl, C3-C6 cycloalkyl, 4-10 membered heterocyclic group, C6-C 10 aryl or 5-10 membered heteroaryl, the C1-C6 alkyl, C3-C6 cycloalkyl, 4-10 membered heterocyclic, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted by R 3a replace; R 3a Selected from halogen, OH, CN, NH2, -COR 3b 、-COOR 3b 、-NHCOR 3b ,-CONHR 3b , -O-C1-C6 alkyl, C1-C6 alkyl, phenyl, 4-6 membered heterocyclyl or 5-6 membered heteroaryl, wherein the C1-C6 alkyl, phenyl, 4-6 membered heterocyclyl or 5-6 membered heteroaryl is optionally replaced by R 3c replace; R 3b Selected from H or C1-C6 alkyl; R 3c Selected from halogen, OH, C1-C6 alkyl or -O-C1-C6 alkyl; R 3’ Selected from H or C1-C6 alkyl; R 4 Selected from NO2, CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl or -O-C1-C6 alkyl; R 5 is selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, -COOC1-C6 alkyl, -S-C1-C6 alkyl, -O-C1-C6 alkyl, -Se-C1-C6 alkyl, -S(O)-C1-C6 alkyl or -S(O)2-C1-C6 alkyl; R 6 is selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or -O-C1-C6 alkyl; n is selected from 1, 2, 3, 4, 5, 6, 7, 8 or 9; Provided that, when Y is selected from OH, n is selected from 1, 2 or 3.

2. The compound of formula (I) according to claim 1, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Y is selected from COOH, -CONH2, -COOR 3 or-CONR 3’ R 3 ; Alternatively, Y is selected from CONH2, -COOR 3 or-CONR 3’ R 3 ; Alternatively, Y is selected from -COOR 3 or-CONR 3’ R 3 ; Alternatively, Y is -COOR 3 ; Alternatively, Y is selected from OH, COOH, -COOCH2CH3, -COOCH3, -COOCH(CH3)2, -COOC(CH3)3, -CONHCH3 or -CONH2; Alternatively, Y is selected from -COOCH2CH3, -COOCH3, -COOCH(CH3)2, -COOC(CH3)3, or -CONHCH3; Alternatively, Y is selected from -COOCH2CH3, -COOCH3, -COOCH(CH3)2 or -COO(CH3)3.

3. The compound of formula (I) according to any one of claims 1 to 2, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R 1 For OH.

4. The compound of formula (I) according to any one of claims 1 to 3, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R 2 is selected from H, C1-C6 alkyl or C1-C6 haloalkyl; or R 2 Selected from C1-C6 alkyl, C1-C6 deuterated alkyl or C1-C6 haloalkyl; Or, R 2 Selected from C1-C6 alkyl or C1-C6 haloalkyl; Or, R 2 Selected from H, CH3 or CF3; Or, R 2 Selected from CH3 or CF3.

5. The compound of formula (I) according to any one of claims 1 to 4, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R 3 is selected from C1-C6 alkyl, C1-C6 deuterated alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclic group, C6-C 10 aryl or 5-6 membered heteroaryl, the C1-C6 alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclic, C6-C 10 Aryl or 5-6 membered heteroaryl is optionally substituted by R 3a replace; Or, R 3 is selected from C1-C6 alkyl or C1-C6 deuterated alkyl, wherein the C1-C6 alkyl is optionally substituted by R 3a replace; Or, R 3 is an unsubstituted C1-C6 alkyl group; Or, R 3 is selected from unsubstituted methyl, unsubstituted ethyl, unsubstituted isopropyl or unsubstituted tert-butyl.

6. The compound of formula (I) according to any one of claims 1 to 5, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R 3a Selected from halogen, OH, CN, NH2, -COR 3b 、-COOR 3b 、-NHCOR 3b ,-CONHR 3b , -O-C1-C6 alkyl or C1-C6 alkyl, wherein the C1-C6 alkyl is optionally replaced by R 3c replace.

7. The compound of formula (I) according to any one of claims 1 to 6, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R 3’ is H or C1-C4 alkyl; or, R 3’ For H.

8. The compound of formula (I) according to any one of claims 1 to 7, or its stereoisomer, or a pharmaceutically acceptable salt thereof, wherein: R 4 is selected from halogen or CN; or, R 4 is selected from F, Cl or CN; or, R 4 For CN.

9. The compound of formula (I) according to any one of claims 1 to 8, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R 5 Selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, -COOC1-C6 alkyl, -S-C1-C6 alkyl or -O-C1-C6 alkyl; Or, R 5 Selected from C1-C6 haloalkyl, -S-C1-C6 alkyl, -O-C1-C6 alkyl, -Se-C1-C6 alkyl, -S(O)-C1-C6 alkyl or -S(O)2-C1-C6 alkyl; Or, R 5 Selected from C1-C3 haloalkyl, -S-C1-C3 alkyl, -O-C1-C3 alkyl or -Se-C1-C3 alkyl; Or, R 5 Selected from C1-C6 haloalkyl, -S-C1-C6 alkyl or -O-C1-C6 alkyl; Or, R 5 Selected from CF3, -S-CH3, -O-CH3, -Se-CH3, -S(O)-CH3 or -S(O)2-CH3; Or, R 5 Selected from CF3, -S-CH3, -O-CH3 or -Se-CH3.

10. The compound of formula (I) according to any one of claims 1 to 9, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R 6 is selected from H or halogen; or, R 6 For H.

11. The compound of formula (I) according to any one of claims 1 to 10, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: n is selected from 1, 2 or 3; Alternatively, n is selected from 1 or 2; Alternatively, n is 2.

12. The compound of formula (I) according to any one of claims 1 to 11, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: X is N.

13. The compound of formula (I) according to any one of claims 1 to 12, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The compound represented by formula (I) or its stereoisomer or its pharmaceutically acceptable salt is selected from the compound represented by formula (II) or its stereoisomer or its pharmaceutically acceptable salt, Among them, X, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , n as defined in any one of claims 1-12.

14. The compound of formula (I) according to claim 1, or its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: The compound represented by formula (I) or its stereoisomer or its pharmaceutically acceptable salt is selected from the following compounds or their stereoisomers or their pharmaceutically acceptable salts:

15. A pharmaceutical composition comprising the compound of formula (I) according to any one of claims 1 to 14 or its stereoisomer or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

16. Use of the compound of formula (I) or its stereoisomer or pharmaceutically acceptable salt as claimed in any one of claims 1 to 14, or the pharmaceutical composition as claimed in claim 15, in the preparation of a medicament for preventing or treating androgen receptor-mediated diseases.

17. A method for preventing or treating a disease mediated by an androgen receptor in a mammal, comprising administering a therapeutically effective amount of a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, or a pharmaceutical composition according to claim 15 to a mammal, preferably a human, in need of such treatment.

18. A compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, or a pharmaceutical composition according to claim 15 for preventing or treating a disease mediated by androgen receptor in a mammal.

19. The use according to claim 16, the method according to claim 17, or the compound of formula (I) or its stereoisomer or pharmaceutically acceptable salt or pharmaceutical composition according to claim 18, wherein: The androgen receptor-mediated disease is selected from androgenic alopecia.

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