Glucose-dependent insulinotropic polypeptide receptor small molecule antagonists and uses thereof

Abstract

The present application provides a class of small molecule antagonists of glucose-dependent insulinotropic polypeptide receptor, the compound or pharmaceutical composition can be used for preventing, treating or reducing the patient GIP receptor antagonist mediated metabolic diseases, such as obesity, type 2 diabetes, pulmonary alveolar proteinosis, non-alcoholic fatty liver disease, atherosclerosis, heart failure, polycystic ovary syndrome, Alzheimer's disease or osteoporosis.

Description

[0001] This application claims the following priority:

[0002] CN202510494501.8, Application Date: April 18, 2025;

[0003] CN202510983770.0, Application Date: 2025.07.16;

[0004] CN202511087824.1, Application Date: 2025.08.04;

[0005] CN202512048134.1, Application Date: 2025.12.31; Technical Field

[0006] This invention belongs to the field of pharmaceuticals, specifically relating to glucose-dependent insulinotropic peptide receptor small molecule antagonists and their applications. Background Technology

[0007] The glucose-dependent insulinotropic peptide (GIP) receptor is a G protein-coupled receptor (GPCR) primarily expressed in various tissues, including the pancreas, intestine, adipose tissue, vascular system, heart, and brain (Nat Rev Endocrinol 2023; 18:201-216). The endogenous ligand of GIPR is the glucose-dependent insulinotropic peptide (GIP), which is secreted by intestinal K cells after eating and plays a role in promoting insulin release, regulating lipid metabolism, and maintaining energy balance.

[0008] Recent studies have revealed that the glucose-dependent insulinotropic peptide (GIP) receptor signaling pathway plays a crucial role in diseases such as obesity, type 2 diabetes mellitus (T2DM), and metabolic syndrome, making it an important target for the treatment of metabolic diseases. GIPR gene knockout mice are resistant to high-fat diet-induced weight gain and exhibit improved insulin sensitivity and lipid profiles (Yamada et al. Diabetes. 2006, 55:S86; and Miyawaki et al. Nature Med. 2002, 8:738-742). Recent data support that loss of function of heterozygous GIP receptors leads to a reduction in human BMI and obesity risk (Akbari et al. Science. 2021, 373: 6550). Small molecule, peptide, and monoclonal antibody GIP receptor antagonist activities have been shown to prevent weight gain and insulin resistance in preclinical obesity models (Nakamura et al. DiabetesMetab Syndr Obes. 2021,14:1095-1105; Yang et al. Mol Metab. 2022, 66: 101638; and Killion et al. Sci. Transl. Med., 2018, 10:eaat3392).

[0009] The combined use of glucose-dependent insulinotropic peptide (GIP) receptor modulators and glucagon-like peptide-1 receptor (GLP-1R) agonists has been extensively studied and proven to deliver significant and remarkable weight loss results (Lu et al. CellRep Med. 2021, 2(5):100263). Several multi-target drugs have entered clinical trials, among which zepbound (a dual GIPR and GLP-1R agonist) was approved by the US FDA on November 8, 2023, for the treatment of hypoglycemia and weight loss. Zepbound demonstrated better weight loss efficacy and fewer side effects in a head-to-head clinical trial with semaglutide (Wegovy). Furthermore, the GLP-1R agonist / GIPR antagonist MariTide (maridebart cafraglutide, AMG 133), used to treat obesity and type 2 diabetes, also achieved positive results in a Phase II clinical trial (NCT05669599).

[0010] Currently, modulators targeting glucose-dependent insulinotropic peptide (GIP) receptors mainly focus on peptide molecules (such as antibodies or GIP analogs), but they suffer from problems such as low oral bioavailability, high production costs, and immunogenicity risks. Small molecule antagonists, due to their structural tunability, convenient oral administration, and lower production costs, have become a more promising development direction. Researchers have already conducted some studies in hopes of finding effective small molecule GIP receptor antagonists. For example, WO2024214038A1 discloses a series of small molecule glucose-dependent insulinotropic peptide receptor antagonists. However, further development of more comprehensive and effective small molecule GIP receptor antagonists is still needed. Summary of the Invention

[0011] The purpose of this invention is to provide a novel class of small molecule compounds with excellent GIP receptor inhibitory activity and pharmacokinetic properties.

[0012] Specifically, the present invention provides a compound, which is a compound of formula (I), or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of a compound of formula (I).

[0013] (I);

[0014] in:

[0015] L1 is , , , , phenyl or 5-6 heteroaryl;

[0016] L2 is a bond or -(CR7R8) n -;

[0017] Each R1 is independently selected from H, D, F, Cl, Br, I, NH2, CN, OH, guanidinyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0018] R2 is -OH, - , -B(OH)2, -C(=O)OW1, -C(=O)NW2W3 or -S(=O)2W4, where W1, W2, W3 and W4 are independently selected from H, D, C respectively. 1-6 Alkyl, C 3-8 cycloalkyl and C 6-10 Aryl, the C 1-6 Alkyl, C 3-8cycloalkyl and C 6-10 The aryl group can be independently and optionally replaced by one, two, or three substituents selected from D, F, Cl, Br, I, NH2, CN, OH, and guanidinyl groups;

[0019] Ring A is , Or C 5-10 Bridged heterocyclic alkyl groups;

[0020] Ring B is C 6-10 Aryl or 5-12 heteroaryl, wherein C 6-10 The aryl and 5-12 heteroaryl groups can be independently and optionally surrounded by 1, 2, 3 or 4 groups selected from D, F, Cl, Br, I, oxo (=O), NH2, CN, OH, guanidinyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 1-6 Substituents of cycloalkyl and 3-6 membered heterocyclic alkyl groups;

[0021] Ring C and ring D are independently selected from phenyl, naphthyl, 5-6 membered heteroaryl, and C64, respectively. 3-6 Monocyclic cycloalkyl, 3-6 membered monocyclic heterocyclic alkyl, C 5-10 Spirocycloalkyl, C 5-10 cycloalkyl, C 5-10 Bridged cycloalkyl, 5-10 membered spirocyclic heterocyclic alkyl, 5-10 membered fused heterocyclic alkyl and 5-10 membered bridged cycloalkyl, wherein the phenyl, naphthyl, 5-6 membered heteroaryl, C 3-6 Monocyclic cycloalkyl, 3-6 membered monocyclic heterocyclic alkyl, C 5-10 Spirocycloalkyl, C 5-10 cycloalkyl, C 5-10 Bridged cycloalkyl, 5-10 membered spirocyclic heterocyclic alkyl, 5-10 membered fused heterocyclic alkyl, and 5-10 membered bridged cycloalkyl can be independently and optionally surrounded by 1, 2, 3, or 4 groups selected from D, F, Cl, Br, I, oxo (=O), NH2, CN, OH, guanidinyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 1-6 Substituents of cycloalkyl and 3-6 membered heterocyclic alkyl groups;

[0022] R3, R4, R5, and R6 are independently selected from H, D, F, Cl, Br, I, CN, guanidino, and C, respectively. 1-6 Alkyl and C 1-6 Alkoxy; the C 1-6 Alkyl and C 1-6The alkoxy group can be optionally and independently substituted by 1, 2, 3, 4 or 5 substituents selected from D, F, Cl, Br, I;

[0023] Each of R7 and R8 is independently selected from H, D, F, Cl, Br, I, OH, and C. 1-6 Alkyl and C 1-6 Alkoxy, the C 1-6 Alkyl and C 1-6 The alkoxy group may be independently and optionally substituted by 1, 2, 3, 4 or 5 substituents selected from H, D, F, Cl, Br, I, oxo (=O), CN, NH2 and OH; or R7 and R8 may optionally form a C with the carbon atom they are attached to. 3-6 cycloalkyl, the C 3-6 The cycloalkyl group may be independently and optionally substituted by one, two or three substituents selected from D, oxo (=O), F, Cl, Br, I, CN, NH2 and OH;

[0024] m can be 0, 1, 2, 3, or 4;

[0025] n is 1, 2, or 3.

[0026] In some embodiments, each R1 is independently selected from H, D, F, Cl, Br, I, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3, and -OCH2CF2CHF2.

[0027] In some implementations, ring A is , , , , , , , or , , or .

[0028] In some implementations, L1 is , , , , phenyl, , , , , , , , , , or .

[0029] In some embodiments, ring B is selected from any of the following groups:

[0030] phenyl, , , , , , , , , , , , , , , , , , , , , , , , , , , , and ;

[0031] The above groups may be independently and optionally replaced by 1, 2 or 3 substituents selected from D, oxo (=O), F, Cl, Br, I, CN, NH2, OH, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3 and -OCH2CF2CHF2.

[0032] In some embodiments, ring C and ring D are independently selected from phenyl, naphthyl, and... , , , , , , , , , , , , , , , , , , , , , , , , , and ;

[0033] The above groups may be independently and optionally replaced by 1, 2 or 3 substituents selected from D, oxo (=O), F, Cl, Br, I, CN, NH2, OH, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3 and -OCH2CF2CHF2.

[0034] In some embodiments, R3, R4, R5, and R6 are independently selected from H, D, F, Cl, Br, I, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3, and -OCH2CF2CHF2;

[0035] In some embodiments, R7 and R8 are independently selected from H, D, F, Cl, Br, I, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3, and -OCH2CF2CHF2; or R7 and R8 may optionally form cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl with the carbon atom they are connected to, wherein the cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl may be independently and optionally substituted by one, two, or three substituents selected from D, F, Cl, Br, and I.

[0036] In some embodiments, it is a compound having one of the following structures or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof:

[0037] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

[0038] The present invention also relates to pharmaceutical compositions comprising a compound of formula (I) of the present invention, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, mediator or combination thereof thereof.

[0039] The present invention also relates to the use of the aforementioned compounds or pharmaceutical compositions thereof in the preparation of medicaments for the prevention, treatment or relief of GIP receptor antagonist-mediated diseases in patients.

[0040] Some of these implementations involve GIP receptor antagonists mediated by diseases such as obesity, type 2 diabetes, pulmonary deposition disease, non-alcoholic steatohepatitis, atherosclerosis, heart failure, polycystic ovary syndrome, Alzheimer's disease, or osteoporosis.

[0041] On the other hand, the present invention relates to methods for the preparation, separation and purification of compounds contained in formula (I).

[0042] The compounds of this invention have a significant inhibitory effect on GIP receptors, high plasma exposure, long half-life, high oral bioavailability, and excellent pharmacokinetic properties.

[0043] Definitions and general terms

[0044] This invention will list in detail the relevant literature for the specific details provided, and the embodiments are accompanied by diagrams of structural and chemical formulas. This invention is intended to cover all options, variations, and equivalents that may be included in the field of prior art as defined in the claims. Those skilled in the art will recognize many similar or equivalent methods and substances described herein that can be applied in the practice of this invention. This invention is by no means limited to the description of methods and substances. Many documents and similar substances distinguish or conflict with this application, including but not limited to the definitions of terms, usages of terms, described techniques, or the scope controlled as described in this application.

[0045] Unless otherwise stated, the following definitions will apply in this invention. For the purposes of this invention, chemical elements are defined according to the periodic table, CAS version, and the Chemical Handbook, 75th Ed, 1994. Furthermore, general principles of organic chemistry are found in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry," by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007; therefore, all content incorporates these references.

[0046] The term "comprising" is an open-ended expression, meaning it includes the contents specified in this invention, but does not exclude other aspects.

[0047] Compounds described herein may optionally be substituted with one or more substituents, such as the general formula compounds of this invention, or the specific examples, subclasses, and classes of compounds included in this invention, as described in the embodiments. It should be understood that the term "optionally substituted" is used interchangeably with the term "substituted or unsubstituted." Generally, the term "optionally," whether preceding the term "substituted," indicates that one or more hydrogen atoms in the given structure are substituted by a specific substituent. Unless otherwise indicated, an optional substituent group may have one substituent substituted at each substituted position of the group. When more than one position in the given structural formula can be substituted by one or more substituents selected from a specific group, the substituents may be substituted at the same or different positions. The substituents described may be, but are not limited to, hydrogen, F, Cl, Br, I, nitro, cyano, oxo (=O), hydroxy, alkyl, hydroxyalkyl, alkylamino, aminoalkyl, haloalkoxy, cycloalkyl, amino, aryl, heterocyclic, heteroaryl, alkenyl, alkynyl, cycloalkyloxy, alkoxy, alkoxyalkyl, haloalkyl, etc.

[0048] The C used in this invention m-n This means that the part has an integer number of carbon atoms within a given range. For example, "C 1-6 The "" indicates that the group can have 1, 2, 3, 4, 5, or 6 carbon atoms. For example, "C 1-3 "" means that the group can have 1 carbon atom, 2 carbon atoms, or 3 carbon atoms.

[0049] The term "heteroatom" refers to a heteroatom or heterogroup (i.e., a group containing a heteroatom), including atoms other than carbon (C) and hydrogen (H) and groups containing such heteroatoms. For example, heteroatoms include, but are not limited to, oxygen, nitrogen, sulfur, phosphorus, silicon, germanium, aluminum, and boron. Specific heteroatoms or heterogroups include, for example: -O-, -S-, -N=, =O, =S, -P(=O)-, -P(=O)2-, -P(=O)O-, -P(=O)2O-, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O)2-, and optionally substituted -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O)2N(H)- or -S(=O)N(H)-. Preferably, the term "heterogeneous" means that the heteroatom or heteroatomic group (i.e., the group containing heteroatoms) is selected from oxygen, nitrogen or sulfur.

[0050] Indicates the link site.

[0051] The term "alkyl" refers to a compound with the general formula C1. n H 2n+1 The alkyl group. This alkyl group can be straight-chain or branched. For example, the term "C 1-6 "Alkyl" refers to an alkyl group containing 1 to 6 carbon atoms (e.g., 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.). Similarly, the alkyl portion (i.e., alkyl) of alkoxy, alkylamino, dialkylamino, alkylsulfonyl, and alkylthio groups has the same definition as above.

[0052] The term "cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of carbon atoms, including monocyclic and bicyclic systems, wherein bicyclic systems include spirocyclic, fused, and bridged rings. The cycloalkyl group comprises C... 3-6 C 3-5 C 3-8 C 6-10 C 4-10 C 4-12 C 4-8 C 4-6 C 4-5 C 5-8 Or C 5-6 Cycloalkyl groups, etc.; they can be monovalent, divalent, or polyvalent. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornel, [2.2.2]bicyclooctane, etc.

[0053] The term "heterocyclic alkyl" refers to a fully saturated cyclic group, including monocyclic and bicyclic systems, wherein bicyclic systems include spirocyclic, fused, and bridged rings. Unless otherwise indicated, the heterocycle is typically a 3- to 16-membered, 3- to 11-membered, 3- to 10-membered, 3- to 7-membered, 3- to 6-membered, or 3- to 5-membered ring containing 1 to 3 heteroatoms independently selected from boron, nitrogen, oxygen, sulfur, silicon, and / or phosphorus (preferably heteroatoms of sulfur, oxygen, and / or nitrogen, preferably 1 or 2 heteroatoms). Examples of 3-membered heterocyclic alkyl groups include, but are not limited to, ethylene oxide, cyclothioethylene, and cycloazoethylene; non-limiting examples of 4-membered heterocyclic alkyl groups include, but are not limited to, acridine, oxadiazolyl, and thiobutylcycloyl; examples of 5-membered heterocyclic alkyl groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, imidazolyl, and tetrahydropyrazolyl; examples of 6-membered heterocyclic alkyl groups include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, morpholinyl, piperazine, 1,4-thiaoxane, 1,4-dioxane, thiomorpholinyl, 1,3-dithiaalkyl, and 1,4-dithiaalkyl; and examples of 7-membered heterocyclic alkyl groups include, but are not limited to, azirheptanyl, oxeheptanyl, and thioheptanyl.

[0054] The term "spirocyclic" refers to a polycyclic group in which rings share a single carbon atom (called a spiro atom) between rings. It may contain 0 or more double or triple bonds and 0 to 5 heteroatoms selected from N, O, S, P, Si, and their oxidation states. Spirocyclic rings are typically 5 to 14-membered, 5 to 12-membered, or 5 to 10-membered. Common spirocyclic rings are tri-spirotri- (representing a three-membered ring followed by a spirotri-membered ring), tri-spirotetra-, tri-spiropenta-, tri-spirohexa-, tetra-spirotetra-, tetra-spirotetra-, tetra-spirotetra-, tetra-spirotetra-, tetra-spirotetra-, tetra-spirotetra-, tetra-spirotetra-, penta-spirotetra-, or ... .

[0055] The term "fused ring" refers to a polycyclic group in which rings share two adjacent ring atoms and a chemical bond. It may contain one or more double or triple bonds and can contain 0 to 5 heteroatoms selected from N, S, O, P, Si, and their oxidation states. Fused rings are typically 5 to 20-membered, 5 to 14-membered, 5 to 12-membered, or 5 to 10-membered rings. Common fused ring types include trifused tetracyclic rings (representing fused rings formed by a three-membered and a four-membered ring; according to IUPC nomenclature, this could be a fused ring with either a three-membered or a four-membered ring as the base ring, and the same applies below), trifused pentacyclic rings, trifused hexacyclic rings, tetrafused tetracyclic rings, tetrafused pentacyclic rings, tetrafused hexacyclic rings, pentafused pentacyclic rings, pentafused hexacyclic rings, and hexafused hexacyclic rings. Non-limiting examples of fused rings include purines, quinolines, isoquinolines, benzopyrans, benzofurans, benzothiophenes, etc. .

[0056] A "bridged ring" refers to two rings that share two non-adjacent ring atoms and may contain one or more double or triple bonds. A bridged ring can contain 0 to 5 heteroatoms selected from N, S, O, P, Si, and their oxidation states. Typically, a bridged ring has 5 to 20, 5 to 14, 5 to 12, or 5 to 10 ring atoms. Non-limiting examples of bridged rings include:

[0057]

[0058] The term "aryl" refers to a monocyclic, bicyclic, or tricyclic carbocyclic system containing 6-14, 6-12, or 6-10 ring atoms, wherein at least one ring is aromatic, and each ring comprises a ring of 3-7 atoms and has one or more attachment sites connected to the remainder of the molecule. Examples of aryl groups include phenyl, naphthyl, anthracene, indanyl, indenyl, etc. , , , wait.

[0059] The term "heteroaryl" refers to an aromatic ring containing 1 to 5 heteroatoms selected from N, O, S, P, Si, and their oxidation states. It can be monocyclic, bicyclic, or polycyclic. Bicyclic or polycyclic rings can be bridged rings, fused rings, spirocyclic rings, and combinations thereof. When bicyclic or polycyclic, it can be a fusion of a heteroaryl group with an aryl group, a fusion of two heteroaryl groups, or a fusion of a heteroaryl group with a cycloalkyl or heterocycloalkyl group, where the heteroaryl group serves as the linking site. "xy-membered heteroaryl" indicates that the total number of ring atoms in the heteroaryl group is x to y. It can be a 5-6-membered heteroaryl group, or a 5-6-membered heteroaryl group fused with other rings (e.g., cycloalkyl, heterocycloalkyl, aromatic rings), where the heteroaromatic ring serves as the linking site. For example, "5-12-membered heteroaryl" indicates that the heteroaryl group serves as the linking site, and the total number of ring atoms is 5-12.

[0060] Examples of heteroaryl groups include, but are not limited to: furanyl, imidazolyl (e.g., N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), isoxazolyl, oxazolyl (e.g., 2-oxazolyl, 4-oxazolyl, 5-oxazolyl), pyrrolyl (e.g., N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl), pyridyl, pyrimidinyl (e.g., 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl), pyridazinyl, thiazolyl, etc. Azolyl (e.g., 2-thiazolyl, 4-thiazolyl, 5-thiazolyl), tetrazolyl (e.g., 5-tetrazolyl), triazolyl, thiophene (e.g., 2-thiophene, 3-thiophene), pyrazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiodiazolyl, 1,3,4-thiodiazolyl, 1,2,5 -Thiodiazolyl, pyrazinyl, 1,3,5-triazinyl; also includes, but is by no means limited to, the following bicyclic compounds: benzimidazolyl, benzofuranyl, benzothiopheneyl, indoleyl (e.g., 2-indoleyl), purinyl, quinolinyl (e.g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), 1,2,3,4-tetrahydroisoquinolinyl, 1,3-benzodioxonelyl, indolinyl, isoquinolinyl (e.g., 1-isoquinolinyl) Quinolinyl, 3-isoquinolinyl or 4-isoquinolinyl), imidazo[1,2-a]pyridyl, pyrazolo[1,5-a]pyridyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, [1,2,4]triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl, [1,2,4]triazolo[1,5-a]pyridyl, , , , , , , , , , , , , , , , , , , , , wait

[0061] When a bond cross-bonds two atoms in a ring (including monocyclic, fused, or spirocyclic rings), this bond can bond with any atom in the ring (including monocyclic, fused, or spirocyclic rings). For example, structural units. This means that the bonds on both sides can be connected to any two different atoms in ring E, ring F, or ring G; for example... This indicates that the bonds on both sides can be connected to any two different atoms on ring E, the central benzene ring, or ring G; further for example... This indicates that the bonds on both sides can be connected to any two different atoms in the four rings of the system.

[0062] As described in this invention, the group "-O-(CR)" m R n ) p The "-" symbol has two connection sites that can be attached to the rest of the molecule, and the connection methods of the two connection sites can be interchanged. For example, formula f represents the group -O-(CR) m R n ) p - It can be attached to the rest of the molecule via the E-terminus or E'-terminus. For example, EO-(CR m R n ) p -E' or E'-O-(CR m R n ) p -E.

[0063]

[0064] Unless otherwise explicitly stated, the descriptive phrases “each and each is independently”, “each and each is independently”, and “each and each is independently” used throughout this document are interchangeable and should be interpreted broadly. They can mean either that the specific options expressed by the same symbols in different groups do not affect each other, or that the specific options expressed by the same symbols in the same group do not affect each other.

[0065] Unless otherwise indicated, the structural formulas described in this invention include all isomers (e.g., enantiomers, diastereomers, geometric isomers, or conformational isomers): for example, R and S configurations containing an asymmetric center, (Z) and (E) isomers of double bonds, and (Z) and (E) conformational isomers. Therefore, any single stereochemical isomer of the compounds of this invention, or a mixture of its enantiomers, diastereomers, geometric isomers, or conformational isomers, is within the scope of this invention.

[0066] Unless otherwise indicated, the structural formulas and compounds described in this invention include all isomers (e.g., enantiomers, diastereomers, geometric isomers, or conformational isomers), nitrides, hydrates, solvates, metabolites, pharmaceutically acceptable salts, and prodrugs. Therefore, compounds of this invention that are individual stereochemical isomers, enantiomers, diastereomers, geometric isomers, conformational isomers, nitrides, hydrates, solvates, metabolites, pharmaceutically acceptable salts, and prodrugs are also within the scope of this invention.

[0067] The structural formulas of the compounds described in this invention include enriched isotopes of one or more different atoms. Examples of isotopes that can be incorporated into the compounds of this invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, respectively, for example... 2 H, 3 H, 13 C 14 C 15 N、 18 O、 17 O、 31 P, 32 P, 35 S, 18 F and 36 Cl. The compounds of the present invention, their prodrugs, and pharmaceutically acceptable salts of said compounds or prodrugs containing the aforementioned isotopes and / or other atoms are all within the scope of the present invention. Certain isotope-labeled compounds of the present invention, such as those doped with radioactive isotopes, are also within the scope of the present invention. 3 H and 14 Those of type C can be used for drug and / or substrate tissue distribution determination. Tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred because they are easy to prepare and detectable. Furthermore, heavier isotopes (e.g., deuterium, i.e., 2 H) substitution can provide certain therapeutic advantages due to increased metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in some cases. The isotopically labeled compounds and their prodrugs used in the methods of this invention can generally be prepared by using readily available isotopically labeled reagents instead of non-isotopically labeled reagents according to compound preparation procedures disclosed in the art.

[0068] "Metabolic product" refers to the product obtained in vivo by the metabolism of a specific compound described in this invention or its pharmaceutically acceptable salt, analogue, or derivative, which exhibits activity similar to that of the compound of formula (I) in vivo or in vitro. A metabolite of a compound can be identified using techniques known in the art, and its activity can be characterized by experimental methods as described in this invention. Such products can be obtained by methods such as oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, or enzymatic cleavage of an administered compound. Accordingly, this invention includes metabolites of compounds, including metabolites produced by sufficient contact of the compounds of this invention with mammals for a period of time.

[0069] The definitions and conventions of stereochemistry used in this invention are generally referenced in the following literature: SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of this invention may contain asymmetric or chiral centers, and therefore exist as different stereoisomers. All stereoisomers of the compounds of this invention, including, but not limited to, diastereomers, enantiomers, transisomers, and mixtures thereof, such as racemic mixtures, constitute a part of this invention. Many organic compounds exist in optically active forms, i.e., they are capable of rotating the plane of plane-polarized light. In describing optically active compounds, the prefixes D, L, or R, S are used to indicate the absolute configuration of the chiral center of the molecule. The prefixes d, l, or (+), (-) are used to name compounds whose plane polarization is rotated. (-) or l indicates the compound is levorotatory, while (+) or d indicates it is dextrorotatory. These stereoisomers have the same chemical structure, but their stereostructures differ. Specific stereoisomers can be enantiomers, and mixtures of isomers are usually called enantiomeric mixtures. A 50:50 enantiomeric mixture is called a racemic mixture or racemate, which may result in a lack of stereoselectivity or stereodirection during chemical reactions. The terms "racemic mixture" and "racemate" refer to a mixture of two equimolar enantiomers that lack optical activity.

[0070] The terms "tautomer" or "tautomerism form" refer to isomers of different energies that can interconvert through a low energy barrier. For example, proton tautomers (i.e., proton-transfer tautomers) include interconversions via proton transfer, such as isomerization between keto-enol and imine-enamine forms. Valence tautomers include interconversions involving the recombination of bonding electrons.

[0071] As used in this invention, "pharmaceutically acceptable salts" refers to the organic and inorganic salts of the compounds of this invention. Pharmaceutically acceptable salts are well-known in the field, as described in the literature: SM Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19, 1977. Salts formed from pharmaceutically acceptable non-toxic acids include, but are not limited to: inorganic acid salts formed by reaction with amino groups, such as hydrochlorides, hydrobromic acids, phosphates, sulfates, and perchlorates; organic acid salts, such as acetates, oxalates, maleates, tartrates, citrates, succinates, and malonates; or salts obtained by other methods described in the literature, such as ion exchange. Other pharmaceutically acceptable salts include adipate, malate, 2-hydroxypropionate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, transbutenedioic acid, glucono-heptahydrate, glyceryl phosphate, gluconate, hemisulfate, heptahydrate, hexanoate, hydroiodate, 2-hydroxy-ethanesulfonate, lacturonate, lactate, laurate, lauryl sulfate, malate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, borate, pectinate, persulfate, 3-phenylpropionate, picrate, pentanoate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Salts obtained by means of appropriate bases include alkali metals, alkaline earth metals, ammonium, and N+ (C 1-4 Salts of alkyl groups (4). This invention also envisions quaternary ammonium salts formed from any compound containing an N group. Water-soluble or oil-soluble or dispersed products can be obtained by quaternization. Alkali metals or alkaline earth metals that can form salts include sodium, lithium, potassium, calcium, magnesium, etc. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts, and amine cations that resist the formation of equilibrium ions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C... 1-8 Sulfonates and aromatic sulfonates.

[0072] In this invention, "hydrate" refers to an associative compound formed when the solvent molecules are water.

[0073] The term "solvent" in this invention refers to an association formed by one or more solvent molecules and the compound of this invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol.

[0074] The term "nitrogen oxide" in this invention refers to an N-oxide formed by oxidizing one or more nitrogen atoms when the compound contains several amine functional groups. Specific examples of N-oxides are N-oxides of tertiary amines or N-oxides of nitrogen-containing heterocyclic nitrogen atoms. The corresponding amines can be treated with oxidizing agents such as hydrogen peroxide or peracids (e.g., peroxycarboxylic acids) to form N-oxides (see Advanced Organic Chemistry, Wiley Interscience, 4th edition, Jerry March, pages). In particular, N-oxides can be prepared using LWDeady's method (Syn.Comm. 1977, 7, 509-514), for example, by reacting the amine compound with m-chloroperoxybenzoic acid (MCPBA) in an inert solvent (e.g., dichloromethane).

[0075] As used in this invention, the term "prodrug" refers to the conversion of a compound into the compound represented by formula (I) in vivo. Such conversion is influenced by the hydrolysis of the prodrug in the blood or its enzymatic conversion into the parent structure in the blood or tissues. The prodrug compounds of this invention can be esters; in existing inventions, esters that can serve as prodrugs include phenyl esters and aliphatic (C) esters. 1-24Esters, acyloxymethyl esters, carbonates, carbamates, and amino acid esters. For example, one compound of this invention contains a hydroxyl group, meaning it can be acylated to yield a prodrug form. Other prodrug forms include phosphate esters, such as those obtained by phosphorylation of a parent compound with a hydroxyl group. For a complete discussion of prodrugs, please refer to the following literature: T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, Vol. 14 of the ACS Symposium Series; Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987; J. Rautio et al, Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270; and SJ Hecker et al, Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry, 2008, 51, 2328-2345.

[0076] Unless otherwise stated herein or the context clearly indicates otherwise, the terms “an,” “a,” “the,” and similar terms used herein, as well as in the context of the invention (especially in the context of the claims), may be interpreted as including both the singular and the plural. Detailed Implementation

[0077] The present invention will be described in detail below with reference to embodiments, but this does not imply any adverse limitation on the invention. The present invention has been described in detail, and specific embodiments thereof have also been disclosed. 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 invention without departing from the spirit and scope thereof.

[0078] Other specific compounds of the present invention can be prepared by referring to the method of Example 1 and in combination with common knowledge known to those skilled in the art.

[0079] Example 1: 4-(6-((1s, 4s)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-1-carbamoyl)pyridin-3-yl)benzoic acid (compound 1)

[0080]

[0081] Step 1: Synthesis of 3-fluoro-4-(prop-1-en-2-yl)aniline

[0082] 4-Bromo-3-fluoroaniline (25 g, 131.57 mmol) was dissolved in a mixed solution of 1,4-dioxane (500 mL) and water (50 mL). Potassium carbonate (54.55 g, 394.70 mmol) and pinacol isopropenylborate (24.32 g, 144.72 mmol) were added, and the mixture was purged with nitrogen three times. Then, 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (1.93 g, 2.63 mmol) was added, and the mixture was purged with nitrogen three times. The mixture was heated to 90°C and stirred for 36 hours. The reaction mixture was cooled to room temperature, filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate (500 mL × 3). The filtrate was collected, diluted with saturated sodium chloride aqueous solution (500 mL), separated, and the aqueous phase was extracted with ethyl acetate (900 mL × 3). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated. The product was purified by silica gel column chromatography to yield 11.5 g.

[0083] LC-MS (ESI) m / z:[M+H] + =152.1;

[0084] Step 2: Synthesis of 3-fluoro-4-isopropylaniline

[0085] Solution 1: Dissolve 10 g of 3-fluoro-4-(prop-1-en-2-yl)aniline in tetrahydrofuran (100 mL) and methanol (100 mL).

[0086] The fixed bed was completely filled with 5% palladium / alumina (WXSC1035, 10.5 g) granular catalyst. The H2 pressure regulator was adjusted to 1.0 MPa, and the H2 flow rate was 50 mL / min.

[0087] Solution 1 was pumped into the fixed bed of a flow reactor at 1 mL / min and flowed at 50.0 °C for 50 min. The reaction mixture was then collected from the reactor output. The reaction solution was concentrated and purified by silica gel column chromatography to obtain 11.1 g of product.

[0088] LC-MS (ESI) m / z:[M+H] + =154.1;

[0089] Step 3: Synthesis of 2-fluoro-4-isocyanate-1-isopropylbenzene

[0090] 3-Fluoro-4-isopropylaniline (4 g, 26.11 mmol) was dissolved in acetonitrile (160 mL), and triethylamine (3.17 g, 31.33 mmol) was added. The mixture was stirred at 20°C for 10 minutes. Then, 1,1-carbonyldiimidazole (4.23 g, 26.11 mmol) was added, and the mixture was stirred at 20°C for 2 hours. The reaction solution was directly concentrated and proceeded to the next step to obtain 4.65 g of crude product (not suitable for long-term storage).

[0091] Step 4: Synthesis of benzyl 4-(6-aminopyridin-3-yl)benzoate

[0092] 5-Bromopyridine-2-amine (3 g, 17.34 mmol) and 4-benzyloxycarbonylphenylboronic acid (4.44 g, 17.34 mmol) were dissolved in dioxane (30 mL) and water (6 mL). 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (0.63 g, 0.87 mmol) and sodium carbonate (3.68 g, 34.68 mmol) were added. After purging the reaction mixture with nitrogen, the mixture was heated to 80 °C and reacted for 2 hours. The reaction mixture was directly concentrated to remove the solvent, and then extracted with water (50 mL) and dichloromethane (50 mL × 3). The combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated to obtain the crude product, which was purified by normal-phase silica gel column chromatography to give 2.8 g of the product.

[0093] LC-MS (ESI) m / z:[M+H]+ =305.1;

[0094] Step 5: Synthesis of 4-(6-((1s,4s)-2-azabicyclo[2.1.1]hexane-1-carbamate)pyridin-3-yl)benzyl benzoate hydrochloride

[0095] (1s,4s)-2-(tert-Butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-1-carboxylic acid (500 mg, 2.20 mmol) was dissolved in dichloromethane (10 mL), pyridine (870 mg, 11.00 mmol) was added, the mixture was purged with nitrogen three times, phosphorus oxychloride (1011 mg, 6.60 mmol) was added, and the mixture was stirred at 20°C for 0.5 h. Then, benzyl 4-(6-aminopyridin-3-yl)benzoate (669 mg, 2.20 mmol) was added to the reaction mixture, and the mixture was stirred at 20°C for 12 h. The reaction mixture was diluted with ethyl acetate (30 mL), and the pH was adjusted to 2-3 with 1 mol hydrochloric acid aqueous solution, and then separated. The organic phase was extracted with 0.5 mol hydrochloric acid aqueous solution (90 mL × 3), and the aqueous phases were collected and combined. The product was concentrated to remove most of the water, then slurried with methanol (60 mL), filtered, and the filtrate was collected. The solvent was removed by concentration, and the product was separated by high-performance liquid chromatography and lyophilized to obtain 310 mg of the product.

[0096] Step 6: Synthesis of benzyl 4-(6-((1s,4s)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-1-carbamoyl)pyridin-3-yl)benzoate

[0097] 4-(6-((1s,4s)-2-azabicyclo[2.1.1]hexane-1-carbamate)pyridin-3-yl)benzyl benzoate hydrochloride (300 mg, 0.67 mmol) was dissolved in tetrahydrofuran (12 mL), and triethylamine (270 mg, 2.67 mmol) was added. The mixture was stirred at 20°C for 10 min, and a solution of 2-fluoro-4-isocyanate-1-isopropylbenzene (180 mg, 1.00 mmol) in tetrahydrofuran (1.8 mL) was added. The mixture was stirred at 20°C for 1 h. The reaction mixture was diluted with water (30 mL) and ethyl acetate (60 mL), and extracted with ethyl acetate (60 mL × 3). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated. The solution was purified by silica gel column chromatography (petroleum ether:ethyl acetate (containing 20% ​​dichloromethane) = 1:4, Rf = 0.3) to obtain 255 mg.

[0098] LC-MS (ESI) m / z:[M+H] + =593.3;

[0099] Step 7: Synthesis of 4-(6-(((1s,4s)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-1-carbamoyl)pyridin-3-yl)benzoic acid

[0100] Palladium on carbon (250 mg, 2.35 mmol, 10% Pd) was added to a nitrogen-purged reaction flask, which was then submerged in methanol (50 mL). Next, benzyl benzoate (250 mg, 0.42 mmol) was added. The mixture was purged three times with hydrogen and stirred at 20°C under a hydrogen atmosphere of 15 Psi for 0.5 hours. The reaction mixture was diluted with tetrahydrofuran (50 mL), filtered through diatomaceous earth, and the filter cake was washed with tetrahydrofuran (150 mL × 3), followed by washing with a dichloromethane / methanol mixture (5 / 1, 150 mL × 3). The filtrate was collected and concentrated to obtain the crude product. The crude product was dissolved in dimethyl sulfoxide (5 mL) and purified by high performance liquid chromatography to obtain 5 mg of the product.

[0101] LC-MS (ESI) m / z:[M+H] + =503.2;

[0102] 1 H NMR (400 MHz, DMSO-d6) δ = 12.96 (s, 1H), 9.86 (s, 1H), 8.98 (s,1H), 8.70 (s, 1H), 8.20 (s, 2H), 8.02 (d, J = 8.4 Hz, 2H), 7.84 (d, J = 8.4Hz, 2H), 7.37 (dd, J = 12.0 Hz, 14.0 Hz, 1H), 7.20 - 7.15 (m, 2H), 3.53 (s,2H), 3.05 (septet, J = 6.8, 1H), 2.80 (s, 1H), 2.14 (s, 2H), 1.74 - 1.73 (m,2H), 1.15 (d, J = 6.8 Hz, 6H).

[0103] Example 2: 4-(6-((1s,4s)-7-((3-fluoro-4-isopropylphenyl)carbamoyl)-7-azabicyclo[2.2.1]heptane-1-carbamoyl)pyridin-3-yl)benzoic acid (compound 2)

[0104]

[0105] Step 1: Synthesis of (1r,4r)-4-((tert-Butoxycarbonyl)amino)cyclohexylmethanesulfonate

[0106] The flask was purged with nitrogen. (1r,4r)-4-hydroxycyclohexyl)carbamate tert-butyl ester (15 g, 55.74 mmol) was placed in the reaction flask, and dichloromethane (150 mL) and dry triethylamine (10.58 g, 83.61 mmol) were added separately. The mixture was purged with nitrogen three times. The mixture was then cooled to 0°C in an ice bath. Methanesulfonyl chloride (9.67 g, 83.61 mmol) was diluted with dichloromethane (30 mL) and added dropwise to the reaction mixture through a constant-pressure dropping funnel (the addition was completed within approximately 1 hour). The reaction mixture was stirred at 20°C for 12 hours. The pH of the reaction mixture was adjusted to be greater than 8 by slowly adding a saturated sodium bicarbonate aqueous solution, and the reaction was quenched by vigorous stirring. The product was then further diluted with dichloromethane (200 mL) and saturated sodium bicarbonate aqueous solution (200 mL), and the pH value was measured (greater than 8). The mixture was then separated, the organic phase was washed with water (200 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the crude product (approximately 12 g). This crude product was purified by normal-phase column chromatography to obtain 6.1 g of product.

[0107] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.65 - 4.60 (m, 1H), 3.48 - 3.47(m, 1H), 3.01 (s, 3H), 2.31 - 2.06 (m, 4H), 1.70 - 1.66 (m, 2H), 1.44 (s,9H), 1.31 - 1.30 (m, 2H).

[0108] Step 2: Synthesis of (1s,4s)-7-azabicyclo[2.2.1]heptane-7-carboxylic acid tert-butyl ester

[0109] (1r,4r)-4-((tert-Butoxycarbonyl)amino)cyclohexyl methanesulfonate (5 g, 23.22 mmol) was dissolved in tetrahydrofuran (20 mL), and then added in two portions (51.1 mL, 51.13 mmol) of 1 mol potassium tert-butoxide solution in tetrahydrofuran (2 h apart). The reaction mixture was stirred at 20 °C for 12 h. The mixture was diluted with 1 mol hydrochloric acid aqueous solution (105 mL) and ethyl acetate (300 mL), separated, and the aqueous phase was extracted three times with ethyl acetate (300 mL). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated. The product was purified by normal-phase column chromatography to give 1.7 g.

[0110] 1H NMR (400 MHz, CHLOROFORM-d) δ = 4.19 (br s, 2H), 1.77 (br d, J =7.6 Hz, 4H), 1.45 (s, 9H), 1.39 (br d, J = 7.2 Hz, 4H).

[0111] Step 3: Synthesis of (1s,4s)-1-formyl-7-azabicyclo[2.2.1]heptane-7-carboxylic acid tert-butyl ester

[0112] (1s,4s)-7-azabicyclo[2.2.1]heptane-7-carboxylic acid tert-butyl ester (1.2 g, 6.08 mmol) was dissolved in tetrahydrofuran (6 mL), cooled to 0 °C, and N,N,N,N-tetramethylethylenediamine (1.06 g, 9.12 mmol) was added, followed by nitrogen purging three times. A 1.3 mol sec-butyllithium solution in cyclohexane (7.1 mL, 9.12 mmol) was added dropwise, maintaining the temperature at 0–5 °C. The mixture was stirred at 0 °C for 1 hour, and N,N-dimethylformamide (0.89 g, 12.17 mmol) was added, followed by stirring at 0 °C for 2 hours. The reaction mixture was quenched with a saturated ammonium chloride aqueous solution (20 mL), diluted with water (50 mL) and ethyl acetate (50 mL), separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was then purified by normal-phase column chromatography to yield 1.0 g of product.

[0113] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 9.95 (s, 1H), 4.31 (s, 1H), 2.01 -1.93 (m, 4H), 1.63 - 1.55 (m, 4H), 1.43 (s, 9H).

[0114] Step 4: Synthesis of (1s,4s)-7-(tert-butoxycarbonyl)-7-azabicyclo[2.2.1]heptane-1-carboxylic acid

[0115] (1s,4s)-1-formyl-7-azabicyclo[2.2.1]heptane-7-carboxylic acid tert-butyl ester (1 g, 2.93 mmol) and 2-methyl-2-butene (2.26 g, 32.23 mmol) were dissolved in tert-butanol (30 mL). Sodium chlorite (1.32 g, 14.65 mmol) and sodium dihydrogen phosphate (3.51 g, 5.15 mmol) were dissolved in water (10 mL) and then added to the reaction solution. The mixture was stirred at 25 °C for 1 hour. The reaction solution was directly extracted three times with saturated brine (10 mL) and ethyl acetate (30 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was slurried with methyl tert-butyl ether (20 mL), filtered, and the filter cake was washed with methyl tert-butyl ether (10 mL). The filtrate was collected, evaporated to dryness to remove the solvent, and 560 mg of the product was obtained.

[0116] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.37 (t, J = 4.4 Hz, 1H), 2.19 -2.18 (m, 2H), 1.94 - 1.88 (m, 4H), 1.54 - 1.49 (m, 3H), 1.46 (s, 9H).

[0117] Step 5: Synthesis of benzyl 4-(6-((1s,4s)-7-azabicyclo[2.2.1]heptane-1-carbamate)pyridin-3-yl)benzoate

[0118] (1s,4s)-7-(tert-Butoxycarbonyl)-7-azabicyclo[2.2.1]heptane-1-carboxylic acid (560 mg, 2.32 mmol) was dissolved in dichloromethane (12 mL), and pyridine (918 mg, 11.61 mmol) and phosphorus oxychloride (1068 mg, 6.97 mmol) were added, respectively. The mixture was stirred at 0 °C for 10 min, and benzyl 4-(6-aminopyridin-3-yl)benzoate (707 mg, 2.32 mmol) was added. The reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was diluted with ethyl acetate (30 mL), and the pH was adjusted to 2-3 with 1 mol hydrochloric acid aqueous solution. The mixture was separated. The organic phase was extracted three times with 0.5 mol hydrochloric acid aqueous solution (90 mL), and the aqueous phase was collected. Most of the water was removed by concentration, and the mixture was purified by high performance liquid chromatography. The fraction was lyophilized to give 270 mg of the product.

[0119] LC-MS (ESI) m / z: [M+H] + =428.2;

[0120] Step 6: Synthesis of benzyl 4-(6-((1s,4s)-7-((3-fluoro-4-isopropylphenyl)carbamoyl)-7-azabicyclo[2.2.1]heptane-1-carbamoyl)pyridin-3-yl)benzoate

[0121] Benzyl benzoate (250 mg, 0.58 mmol) of 4-(6-((1s,4s)-7-azabicyclo[2.2.1]heptane-1-carbamate)pyridin-3-yl)benzoate was dissolved in tetrahydrofuran (5 mL), and triethylamine (178 mg, 1.75 mmol) was added. The mixture was stirred at 20 °C for 10 min. Then, 2-fluoro-4-isocyano-1-(prop-2-yl)benzene (158 mg, 0.88 mmol) was added, and the reaction mixture was stirred at 20 °C for 2 h. The reaction mixture was diluted with water (30 mL) and ethyl acetate (30 mL), and after separation, the aqueous phase was extracted three times with ethyl acetate (90 mL). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated. The product was purified by normal-phase column chromatography (petroleum ether:ethyl acetate = 2:3, Rf = 0.3) to give 320 mg of product.

[0122] LC-MS (ESI) m / z: [M+H] + =607.4;

[0123] Step 7: Synthesis of 4-(6-((1s,4s)-7-((3-fluoro-4-isopropylphenyl)carbamoyl)-7-azabicyclo[2.2.1]heptane-1-carbamoyl)pyridin-3-yl)benzoic acid

[0124] Palladium on carbon (300 mg, 2.82 mmol, 10% Pd) was added to a nitrogen-purged reaction flask, and the flask was submerged in methanol (30 mL) and tetrahydrofuran (10 mL). Benzyl benzoate (310 mg, 0.51 mmol) was added to the reaction mixture, and the mixture was purged with hydrogen three times. The reaction mixture was stirred at 20°C for 30 minutes under a hydrogen atmosphere (15 Psi). The reaction mixture was filtered, and the filter cake was washed with tetrahydrofuran (150 mL). Finally, the mixture was washed three times with a dichloromethane / methanol (5 / 1) mixture (150 mL). The filtrate was collected and concentrated to obtain the crude product. The crude product was purified by high performance liquid chromatography, and the fraction was freeze-dried to obtain 80 mg of product.

[0125] LC-MS (ESI) m / z: [M+H] + =517.3;

[0126] 1 H NMR (400 MHz, DMSO-d6) δ = 13.00 (br s, 1H), 9.53 (s, 1H), 9.28 (s,1H), 8.68 (s, 1H), 8.21 (d, J = 8.0 Hz, 2H), 8.02 (d, J = 8.4 Hz, 2H), 7.85(d, J = 8.4 Hz, 2H), 7.33 (d, J = 14.4 Hz, 1H), 7.20 - 7.16 (m, 2H), 4.58 (t,J = 4.4 Hz, 1H), 3.06 -3.03 (m, 1H), 2.18 (br t, J = 11.2 Hz, 2H), 1.88 -1.79 (m, 4H), 1.62 (br t, J = 4.0, 2H), 1.15 (d, J = 6.8, 6H).

[0127] Example 3: 4'-((1s,4s)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (compound 3)

[0128]

[0129] Step 1: Synthesis of methyl 4'-amino-[1,1'-biphenyl]-4-carboxylic acid hydrochloride

[0130] 4-(4-aminophenyl)benzoic acid (5.00 g, 23.45 mmol) was dissolved in methanol (50 mL), cooled to 0 °C, and thionyl chloride (5.58 g, 46.90 mmol) was added. The mixture was stirred at 20 °C for 20 minutes, then heated to 60 °C and stirred for 3 hours. The reaction solution was filtered directly, and the filter cake was washed with methyl ether (30 mL). The filter cake was collected, the solvent was removed, and 4.0 g of the product was obtained.

[0131] Step 2: Synthesis of methyl 4'-(((1s,4s)-2-azabicyclo[2.1.1]hexane-1-formamido)-[1,1'-biphenyl]-4-carboxylic acid

[0132] (1s,4s)-2-(tert-Butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-1-carboxylic acid (500 mg, 2.20 mmol) was dissolved in dichloromethane (10 mL), and pyridine (870 mg, 11.00 mmol) was added. After purging with nitrogen three times, phosphorus oxychloride (1012 mg, 6.60 mmol) was added. The reaction mixture was stirred at 20 °C for 1 hour. Then, methyl 4'-amino-[1,1'-biphenyl]-4-carboxylic acid hydrochloride (670 mg, 2.20 mmol) was added to the reaction mixture. The mixture was stirred at 20 °C for 12 hours. The reaction mixture was diluted with ethyl acetate (30 mL), and the reaction was quenched with water. The mixture was separated, and the organic phase was extracted three times with 0.5 mol / L hydrochloric acid aqueous solution (90 mL). The aqueous phase was collected and concentrated to remove water. The product (1.0 g) was purified by normal-phase column chromatography.

[0133] LC-MS (ESI) m / z: [M+H] + =337.2;

[0134] Step 3: Synthesis of 4'-(((1s,4s)-2-azabicyclo[2.1.1]hexane-1-formamido)-[1,1'-biphenyl]-4-carboxylic acid

[0135] Methyl 4'-(((1s,4s)-2-azabicyclo[2.1.1]hexane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (1 g, 2.97 mmol) was dissolved in acetonitrile (30 mL), and potassium trimethylsilanol (2.29 g, 17.84 mmol) was added. The reaction mixture was stirred at 20 °C for 12 hours. Subsequent analysis showed that 10% of the starting material remained, so the reaction mixture was heated to 70 °C and stirred for 1 hour. The reaction mixture was concentrated to remove the solvent, and methanol (30 mL) was added and stirred. The mixture was then filtered. The filtrate was collected and concentrated to obtain the crude product. The crude product was purified by high-performance liquid chromatography, and the fraction was lyophilized to obtain 320 mg of the product.

[0136] LC-MS (ESI) m / z: [M+H] + =323.1;

[0137] Step 4: Synthesis of 4'-(((1s,4s)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0138] 4'-(((1s,4s)-2-azabicyclo[2.1.1]hexane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (200 mg, 0.62 mmol) was dissolved in tetrahydrofuran (10 mL), and triethylamine (251 mg, 2.48 mmol) was added. The reaction mixture was stirred at 25 °C for 10 min, and then 2-fluoro-4-isocyanate-1-isopropylbenzene (222 mg, 1.24 mmol) was added. The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was quenched with methanol and concentrated. The product was purified by high performance liquid chromatography and lyophilized to give 90 mg of product.

[0139] LC-MS (ESI) m / z: [M+H] + =502.3;

[0140] 1 H NMR (400 MHz, DMSO-d6) δ = 12.84 (br s, 1H), 9.60 (s, 1H), 8.92 (s,1H), 7.99 (d, J = 8.0 Hz, 2H), 7.83 - 7.77 (m, 4H), 7.69 (d, J = 8.4 Hz, 2H),7.37 (d, J = 13.6 Hz, 1H), 7.20 - 7.19 (m, 2H), 3.54 (s, 2H), 3.09-3.02 (m,1H), 2.81 (br s, 1H), 2.12 (s, 2H), 1.72-1.67 (m, 2H), 1.16 (d, J = 7.2 Hz, 6H).

[0141] Example 4: 4'-((1S,3R,4R)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.2.1]heptane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (compound 4)

[0142]

[0143] Step 1: Synthesis of (1S,3R,4R)-3-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-2-azabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester

[0144] (1S,3R,4R)-2-(tert-butoxycarbonyl)-2-azabicyclo[2.2.1]heptane-3-carboxylic acid (250 mg, 1.04 mmol) was dissolved in dichloromethane (5 mL), and methyl 4'-amino-[1,1'-biphenyl]-4-carboxylic acid (236 mg, 1.04 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (239 mg, 1.24 mmol) were added. The reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was diluted with water (20 mL) and ethyl acetate (20 mL), separated, and the aqueous phase was extracted three times with ethyl acetate (30 mL). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated to remove the solvent. The crude product was purified by normal-phase column chromatography, and the fraction was concentrated to remove the solvent, yielding 450 mg of product.

[0145] LC-MS (ESI) m / z: [M+H] + =451.2;

[0146] Step 2: Synthesis of 4'-((1S,3R,4R)-2-(tert-butoxycarbonyl)-2-azabicyclo[2.2.1]heptane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0147] (1S,3R,4R)-3-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-2-azabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester (250 mg, 0.55 mmol) was dissolved in acetonitrile (10 mL), and potassium trimethylsilanolate (214 mg, 1.66 mmol) was added. The reaction mixture was stirred at 20°C for 12 hours. When the reaction mixture showed any remaining starting material, it was heated to 60°C and stirred for 1 hour. The pH was adjusted to 6-7 by adding 1 mol of dilute hydrochloric acid aqueous solution, and then diluted with water (50 mL) and ethyl acetate (50 mL). The mixture was separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to remove the solvent, yielding 226 mg of the product.

[0148] LC-MS (ESI) m / z: [M+H] + =437.1;

[0149] Step 3: Synthesis of 4'-(((1S,3R,4R)-2-azabicyclo[2.2.1]heptane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid trifluoroacetate

[0150] 4'-((1S,3R,4R)-2-(tert-butoxycarbonyl)-2-azabicyclo[2.2.1]heptane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (250 mg, 0.55 mmol) was dissolved in dichloromethane (4 mL), and trifluoroacetic acid (2 mL) was added. The mixture was stirred at 25°C for 1 hour. The reaction solution was diluted with dichloromethane (20 mL), concentrated, and the solvent was removed to give 200 mg of the product.

[0151] Step 4: Synthesis of 4'-((1S,3R,4R)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.2.1]heptane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0152] 4'-(((1S,3R,4R)-2-azabicyclo[2.2.1]heptane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid trifluoroacetate (150 mg, 0.33 mmol) was dissolved in tetrahydrofuran (6 mL), and triethylamine (226 mg, 2.23 mmol) was added with stirring, followed by 2-fluoro-4-isocyanate-1-isopropylbenzene (160 mg, 0.89 mmol). The mixture was stirred at 20°C for 2 hours. The reaction solution was quenched with methanol (5 mL), the solvent was removed by concentration, and the crude product was separated by high performance liquid chromatography and lyophilized to obtain 122 mg of the product.

[0153] LC-MS (ESI) m / z: [M+H] + =516.2;

[0154] 1H NMR (400 MHz, DMSO-d6) δ = 12.94 (s, 1H), 10.07 (s, 1H), 8.52 (s,1H), 7.99 (d, J = 8.4 Hz, 2H), 7.79 - 7.70 (m, 6H), 7.43 (dd, J = 2.0, 14.0Hz, 1H), 7.25 (dd, J = 2.0, 8.4 Hz, 1H), 7.17 (t, J = 8.8 Hz, 1H), 4.60 (s,1H), 4.00 (s, 1H), 3.08 - 3.05 (m, 1H), 2.68 (s, 1H), 1.99 (br d, J = 8.8 Hz,1H), 1.77 - 1.61 (m, 3H), 1.39 - 1.37 (m, 1H), 1.38 (br d, J = 9.6 Hz, 1H), 1.17 (d, J = 6.8 Hz, 6H).

[0155] Example 5: 4'-(((1S,3R,4S)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid, 4'-((1R,3S,4R)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid (compounds 5A and 5B)

[0156]

[0157] Step 1: Synthesis of 3-formyl-2-azabicyclotert-butyl[2.1.1]hexane-2-carboxylic acid ester

[0158] The starting material, 2-azabicyclotert-butyl[2.1.1]hexane-2-carboxylate (500 mg, 2.73 mmol), was dissolved in anhydrous tetrahydrofuran (5 mL). The solution was cooled to -70 °C, and 2,5-dimethyl-2,5-diazahexane (476 mg, 4.09 mmol) was added. The mixture was purged with nitrogen three times, and a 1.4 mol solution of sec-butyllithium in tetrahydrofuran (3.2 mL, 4.09 mmol) was added dropwise. The reaction mixture was kept at -70 ± 10 °C and stirred at -70 °C for 2 hours. Then, anhydrous N,N-dimethylformamide (399 mg, 5.46 mmol) was added, and the reaction mixture was heated to 0–25 °C and stirred for 0.5 hours. The reaction mixture was quenched with a saturated ammonium chloride aqueous solution (20 mL), diluted with water (50 mL) and ethyl acetate (50 mL), and separated. The aqueous phase was extracted three times with ethyl acetate (150 mL). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated to remove the solvent. The crude product was purified by normal-phase column chromatography, and the fraction was concentrated to remove the solvent, yielding 120 mg of product.

[0159] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 9.75 (br s, 1H), 4.46 (br s, 1H), 4.03 (br s, 1H), 3.03 - 3.02 (m, 1H), 2.07 (br d, J = 6.4 Hz, 1H), 1.91 (brd, J = 8.0 Hz, 1H), 1.67 - 1.62 (m, 2H), 1.47 (s, 9H).

[0160] Step 2: Synthesis of 2-(tert-Butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-3-carboxylic acid

[0161] The starting materials 3-formyl-2-azabicyclotert-butyl[2.1.1]hexane-2-carboxylic acid ester (120.00 mg, 0.57 mmol) and 2-methyl-2-butene (439 mg, 6.25 mmol) were dissolved in tert-butanol (3 mL), and a prepared solution of sodium chlorite (256 mg, 2.84 mmol) and sodium dihydrogen phosphate (618 mg, 5.15 mmol) in water (1 mL) was added. The reaction solution was stirred at 25 °C for 1 hour. The reaction solution was directly diluted with saturated brine (10 mL) and extracted three times with ethyl acetate (30 mL). The organic phase was dried and filtered, and the filtrate was directly concentrated to obtain the crude product. The crude product was added to methyl tert-butyl ether (20 mL) and stirred, filtered, and the filter cake was washed with methyl tert-butyl ether (10 mL). The filtrate was collected, and the solvent was removed to obtain 110 mg of product.

[0162] H NMR (400 MHz, CHLOROFORM-d) δ = 4.34 (d, J = 6.8 Hz, 1H), 4.25 (s,1H), 3.14 - 3.13 (m, 1H), 2.06 (br d, J = 6.4 Hz, 1H), 2.00 (br d, J = 6.8Hz, 1H), 1.67 - 1.61 (m, 1H), 1.53 - 1.45 (m, 10H).

[0163] Step 3: Synthesis of 3-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-2-azabicyclo[2.1.1]hexane-2-carboxylic acid tert-butyl ester

[0164] The starting material 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-3-carboxylic acid (110 mg, 0.48 mmol) was dissolved in dichloromethane (3 mL), and methyl 4'-amino-[1,1'-biphenyl]-4-carboxylic acid (110 mg, 0.48 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (111 mg, 0.58 mmol) were added. The reaction mixture was stirred at 25 °C for 12 hours. The mixture was diluted with water (20 mL) and ethyl acetate (60 mL), separated, and the aqueous phase was extracted three times with ethyl acetate (120 mL). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by normal-phase column chromatography (petroleum ether:ethyl acetate (containing 20% ​​dichloromethane) = 2:1, Rf = 0.4), and the fraction was concentrated to remove the solvent, yielding 130 mg of the product.

[0165] Step 4: Synthesis of 4'-(2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0166] The starting material 3-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-2-azabicyclo[2.1.1]hexane-2-carboxylic acid tert-butyl ester (130 mg, 0.30 mmol) was dissolved in acetonitrile (10 mL), and potassium trimethylsilanolate (191.04 mg, 1.49 mmol) was added. The reaction mixture was stirred at 25 °C for 12 hours. The solution was diluted with water (50 mL) and ethyl acetate (100 mL), and then the pH was adjusted to 3-4 with 1 mol of dilute hydrochloric acid aqueous solution. The mixture was separated, and the aqueous phase was extracted three times with ethyl acetate (300 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to remove the solvent, yielding 120 mg of the product.

[0167] Step 5: Synthesis of 4'-(2-azabicyclo[2.1.1]hexane-3-formamido)-[1,1'-biphenyl]-4-carboxylic acid trifluoroacetate

[0168] The starting material 4'-(2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (120 mg, 0.28 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at 25 °C for 1 hour. After dilution with dichloromethane (10 mL), the solution was concentrated to remove the solvent, yielding 110 mg of the product.

[0169] Step 6: Synthesis of 4'-(2-(((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0170] 4'-(2-azabicyclo[2.1.1]hexane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid trifluoroacetate (91 mg, 0.21 mmol) was dissolved in tetrahydrofuran (5 mL), and triethylamine (143 mg, 1.41 mmol) was added. The mixture was stirred at 25 °C for 5 minutes, followed by the addition of 2-fluoro-4-isocyanate-1-isopropylbenzene (102 mg, 0.56 mmol). The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was quenched with methanol (5 mL), and the solvent was removed by concentration. The crude product was separated by high performance liquid chromatography, and the fraction was lyophilized to obtain 95 mg of the product.

[0171] LC-MS (ESI) m / z: [M+H] + =502.3;

[0172] Step 7: Synthesis of 4'-(((1S,3R,4S)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid and 4'-((1R,3S,4R)-2-((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid

[0173] The racemic mixture 4'-(2-(((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (90 mg, 0.18 mmol) was chirally resolved, and the fraction was lyophilized to give compound 5A: 27 mg, retention time 1.704 min; and compound 5B: 40 mg, retention time 1.981 min.

[0174] Compound 5A:

[0175] LC-MS (ESI): [M+H] + =502.2;

[0176] 1 H NMR (400 MHz, DMSO-d6) δ = 12.90 (br s, 1H) 10.21 (s, 1H), 8.76 (s,1H), 7.99 (d, J = 8.4 Hz, 2H), 7.80 - 7.76 (m, 4H), 7.72 (d, J = 8.8 Hz, 2H), 7.45 (dd, J = 2.0, 13.2 Hz, 1H), 7.25 (br d, J = 8.4 Hz, 1H), 7.18 (t, J =8.8 Hz, 1H), 4.74 (br d, J = 6.8 Hz, 1H), 4.44 (s, 1H), 3.10 - 3.04 (m, 2H),2.11 - 2.10 (m, 1H), 1.97 - 1.95 (m, 1H), 1.83 (t, J = 8.0 Hz, 1H), 1.42 (t,J = 8.4 Hz, 1H), 1.18 (d, J = 6.8 Hz, 6H).

[0177] Compound 5B:

[0178] LC-MS (ESI): [M+H] + =502.3

[0179] 1H NMR (400 MHz, DMSO-d6) δ = 12.90 (br s, 1H), 10.20 (s, 1H), 8.76(s, 1H), 7.99 (d, J = 8.4 Hz, 2H), 7.80 - 7.76 (m, 4H), 7.72 (d, J = 8.8 Hz,2H), 7.44 (dd, J = 2.0, 13.6 Hz, 1H), 7.23 (br dd, J =2.0, 8.4 Hz, 1H), 7.16(br d, J = 8.8 Hz, 1H), 4.74 (br d, J = 7.0 Hz, 1H), 4.44 (s, 1H), 3.10 -3.04 (m, 2H), 2.11 - 2.10 (m, 1H), 1.96 - 1.95 (m, 1H), 1.86 (t, J = 10.4 Hz, 1H), 1.43 - 1.42 (m, 1H), 1.18 (d, J = 6.8 Hz, 6H).

[0180] Example 6: (R)-4'-(5-(((3-fluoro-4-isopropylphenyl)carbamoyl)-5-azaspiro[2.3]hexane-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid, (S)-4'-(5-(((3-fluoro-4-isopropylphenyl)carbamoyl)-5-azaspiro[2.3]hexane-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (compounds 6A and 6B)

[0181]

[0182] Step 1: Synthesis of 4-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-5-azaspiro[2,3]hexane-5-carboxylic acid tert-butyl ester

[0183] The starting material 5-(tert-butoxycarbonyl)-5-azaspiro[2,3]hexane-4-carboxylic acid (100 mg, 0.44 mmol) was dissolved in dichloromethane (5 mL), and methyl 4'-amino-[1,1'-biphenyl]-4-carboxylic acid (99 mg, 0.44 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (106 mg, 0.55 mmol) were added separately. The reaction mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with water (60 mL) and ethyl acetate (60 mL), separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated. The product was purified by normal-phase column chromatography to give 190 mg.

[0184] LC-MS (ESI) m / z: [M+Na] + =459.2;

[0185] Step 2: Synthesis of 4'-(5-(tert-butoxycarbonyl)-5-azaspiro[2.3]hexane-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0186] The starting material 4-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-5-azaspiro[2,3]hexane-5-carboxylic acid tert-butyl ester (190 mg, 0.44 mmol) was dissolved in acetonitrile (10 mL), and potassium trimethylsilanolate (223 mg, 1.74 mmol) was added. The reaction mixture was stirred at 25 °C for 4 hours. The pH of the reaction mixture was adjusted to 3-4 by adding 1 mol of dilute hydrochloric acid aqueous solution, and then diluted with ethyl acetate (50 mL). The mixture was separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to remove the solvent, yielding 190 mg of the product.

[0187] LC-MS (ESI) m / z: [M+Na] + =445.2;

[0188] Step 3: Synthesis of 4'-(5-azaspiro[2.3]hexane-4-formamido)-[1,1'-biphenyl]-4-carboxylic acid trifluoroacetate

[0189] The starting material 4'-(5-(tert-butoxycarbonyl)-5-azaspiro[2.3]hexane-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (170 mg, 0.40 mmol) was dissolved in dichloromethane (17 mL), and trifluoroacetic acid (1.5 mL) was added. The reaction mixture was stirred at 25 °C for 0.5 hours. The reaction mixture was diluted with dichloromethane, concentrated, and the solvent was removed to give 170 mg of the product.

[0190] Step 4: Synthesis of 4'-(5-(((3-fluoro-4-isopropylphenyl)carbamoyl)-5-azaspiro[2.3]hexane-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0191] The starting material 4'-(5-azaspiro[2.3]hexane-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid trifluoroacetate (170 mg, 0.39 mmol) was dissolved in tetrahydrofuran (10 mL), and triethylamine (200 mg, 1.98 mmol) was added. The mixture was stirred at 25 °C for 5 minutes, and then 2-fluoro-4-isocyanate-1-isopropylbenzene (180 mg, 1.00 mmol) was added to the reaction solution. The reaction solution was stirred at 25 °C for 4 hours. The reaction solution was quenched with methanol (5 mL), and the solvent was removed by concentration. The crude product was separated by high performance liquid chromatography, and the fraction was lyophilized to obtain 120 mg of product.

[0192] LC-MS (ESI) m / z: [M+H] + =502.3;

[0193] Step 5: Synthesis of (R)-4'-(5-(((3-fluoro-4-isopropylphenyl)carbamoyl)-5-azaspiro[2.3]hexane-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid and (S)-4'-(5-(((3-fluoro-4-isopropylphenyl)carbamoyl)-5-azaspiro[2.3]hexane-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0194] The starting material 4'-(5-(((3-fluoro-4-isopropylphenyl)carbamoyl)-5-azaspiro[2.3]hexane-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (120 mg, 0.25 mmol) was resolved by SFC. Fraction 1 was concentrated to remove the solvent, yielding compound 6A: 45 mg, retention time 1.924 min; Fraction 2 was concentrated to remove the solvent, yielding compound 6B: 40 mg, retention time 2.479 min.

[0195] Compound 6A: LC-MS (ESI): [M+H] + =502.3;

[0196] 1H NMR (400 MHz, DMSO-d6) δ = 9.95 (s, 1H), 8.78 (s, 1H), 8.00 (d, J =8.0 Hz, 2H), 7.77 (d, J = 8.8 Hz, 4H), 7.72 (d, J = 8.8 Hz, 2H), 7.43 (dd, J= 1.6, 13.6 Hz, 2H), 7.22 - 7.18 (m, 2H), 4.74 (s, 1H), 4.16 (d, J = 7.2 Hz,1H), 4.06 (d, J = 7.6 Hz, 1H), 3.07 (q, J = 6.8 Hz, 1H), 1.18 (d, J = 6.8 Hz, 6H), 0.86 - 0.78 (m, 2H), 0.62 - 0.54 (m, 2H).

[0197] Compound 6B:

[0198] LC-MS (ESI): [M+H] + =502.2;

[0199] 1 H NMR (400 MHz, DMSO-d6) δ = 9.96 (s, 1H), 8.79 (s, 1H), 8.00 (d, J= 8.0 Hz, 2H), 7.78 (d, J = 8.8 Hz, 4H), 7.72 (d, J = 8.8 Hz, 2H), 7.43 (dd,J = 1.6, 13.6 Hz, 2H), 7.22 - 7.18 (m, 2H), 4.74 (s, 1H), 4.17 (d, J = 7.6Hz, 1H), 4.06 (d, J = 7.6 Hz, 1H), 3.07 (q, J = 7.2 Hz, 1H), 1.18 (d, J = 6.8Hz, 6H), 0.86 - 0.77 (m, 2H), 0.62 - 0.54 (m, 2H).

[0200] Example 7: 8-((1R,2R,5S)-3-((4-cyclobutylphenyl)carbamoyl)-3-azabicyclo[3.1.0]hexane-2-carboxamido)-6H-benzo[c]chromene-3-carboxylic acid (Compound 7)

[0201]

[0202] Step 1: Synthesis of (Z)-4-((tert-butyldiphenylsilyl)oxy)butyl-2-en-1-ol

[0203] (2Z)-But-2-en-1,4-diol (5 g, 56.75 mmol) was dissolved in tetrahydrofuran (50 mL), cooled to 0 °C, and then sodium hydride (2.27 g, 56.75 mmol) was added. The reaction mixture was stirred at 0 °C for 30 minutes, and then chloro(2-methylpropyl-2-yl)diphenylsilane (14.7 mL, 56.75 mmol) was added. The reaction mixture was naturally heated to 25 °C and stirred for 16 hours. The reaction mixture was quenched with saturated ammonium chloride aqueous solution and extracted with ethyl acetate (200 mL × 3). The combined organic phases were washed once with saturated brine (500 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated to obtain a crude product, which was purified by normal-phase silica gel column chromatography to yield 16.8 g of the product.

[0204] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 7.71 (br d, J = 7.2 Hz, 4H), 7.52- 7.36 (m, 6H), 5.81 - 5.59 (m, 2H), 4.29 (d, J = 5.8 Hz, 2H), 4.08 - 3.98(m, 2H), 1.56 - 1.50 (m, 1H), 1.08 (d, J = 1.0 Hz, 9H).

[0205] Step 2: Synthesis of (3-(((tert-butyldiphenylsilyl)oxy)methyl)ethylene oxide-2-yl)methanol

[0206] (Z)-4-((tert-butyldiphenylsilyl)oxy)butyl-2-en-1-ol (15 g, 45.94 mmol) was dissolved in dichloromethane (200 mL), and m-chloroperoxybenzoic acid (24.1 mL, 66.61 mmol) was slowly added at 0 °C. The reaction mixture was stirred at 20 °C for 18 hours. The reaction mixture was poured into ice water (200 mL) and extracted with dichloromethane (200 mL × 3). The extract was first washed with saturated sodium sulfite aqueous solution, and then the organic phases were combined and washed with saturated brine (200 mL). The mixture was dried over anhydrous sodium sulfate and filtered to obtain the crude product. The crude product was purified by normal-phase silica gel column chromatography to obtain 13 g of the product.

[0207] LC-MS (ESI): [M+Na] + =365.1;

[0208] Step 3: Synthesis of tert-butyl(3-(chloromethyl)epoxyethylene-2-yl)methoxy)diphenylsilane

[0209] (3-(((tert-butyldiphenylsilyl)oxy)methyl)ethylene oxide-2-yl)methanol (7.9 g, 23.07 mmol) was dissolved in carbon tetrachloride (60 mL), and triphenylphosphine (9.1 g, 34.60 mmol) and sodium bicarbonate (4.2 g, 50.74 mmol) were added. The reaction mixture was stirred at 80 °C for 5 hours. The reaction mixture was poured into water (100 mL), and extracted with ethyl acetate (120 mL × 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered to obtain the crude product. The crude product was purified by normal-phase silica gel column chromatography to obtain 7.5 g of the product.

[0210] LC-MS (ESI): [M+H] + =361.1;

[0211] Step 4: Synthesis of 1-((tert-butyldiphenylsilyl)oxy)but-3-yne-2-ol

[0212] tert-Butyl(3-(chloromethyl)epoxyethylene-2-yl)methoxy)diphenylsilane (7 g, 19.39 mmol) was dissolved in tetrahydrofuran (90 mL), and n-butyllithium (23.3 mL, 58.18 mmol) was added at -78 °C. The reaction mixture was stirred at -78 °C for 1 hour. The reaction mixture was poured into an ice-cold ammonium chloride solution (200 mL), and extracted with ethyl acetate (300 mL × 3). The combined organic phases were washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, and filtered to obtain the crude product. The crude product was purified by normal-phase silica gel column chromatography to give 2.8 g of the product.

[0213] LC-MS (ESI): [MH] - =323.0;

[0214] Step 5: Synthesis of 4-methyl-N-(prop-2-yn-1-yl)benzenesulfonamide

[0215] Proprynne-1-amine (11.6 mL, 181.55 mmol) and triethylamine (25.2 mL, 181.55 mmol) were dissolved in dichloromethane (100 mL), and 4-methylbenzenesulfonyl chloride (32.9 mL, 172.48 mmol) was added. The reaction mixture was stirred at 25 °C for 2 hours. Ethyl acetate (500 mL) was added to the reaction mixture, and the organic phase was washed once with a brine solution (500 mL). The organic phase was washed once with saturated brine (500 mL) and filtered. The filtrate was directly concentrated to give 32 g of product.

[0216] LC-MS (ESI): [M+H] + =210.1;

[0217] Step 6: Synthesis of N-(3-bromoprop-2-yn-1-yl)-4-methylbenzenesulfonamide

[0218] 4-Methyl-N-(prop-2-ynyl)benzenesulfonamide (5.2 g, 24.85 mmol) was dissolved in acetone (40 mL), cooled to 0 °C, and then silver nitrate (0.1 mL, 2.48 mmol) and N-bromosuccinimide (4.42 g, 24.85 mmol) were added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was filtered directly through a filter cake lined with diatomaceous earth, and the filter cake was washed with ethyl acetate (50 mL). The combined organic phases were evaporated to dryness, and the crude product was purified by silica gel column chromatography to give 4.8 g of the product.

[0219] LC-MS (ESI): [M+H] + =290.0;

[0220] Step 7: Synthesis of N-(3-(tert-butylthio)prop-2-yn-1-yl)-4-methylbenzenesulfonamide

[0221] N-(3-bromoprop-2-yn-1-yl)-4-methylbenzenesulfonamide (4.8 g, 16.66 mmol), 4-(2-methylprop-2-yl)-2-[4-(2-methylprop-2-yl)pyridin-2-yl]pyridine (0.9 mL, 3.33 mmol) and 2,6-dimethylpyridine (3.9 mL, 33.31 mmol) were dissolved in acetonitrile (60 mL), purged with nitrogen three times, and then 2-methylprop-2-thiol (2.1 mL, 18.32 mmol) and copper(1+)hexafluoro-λ5-phosphonotetra(acetonitrile) (0.6 g, 1.67 mmol) were added. The reaction mixture was stirred at 20 °C for 1 hour. The reaction solution was poured into water (200 mL), and ethyl acetate (200 mL × 3) was added for extraction. The organic phases were combined and washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was evaporated to dryness to obtain the crude product. The crude product was purified by normal phase extraction to obtain 3.7 g of the product.

[0222] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 7.77 (d, J = 8.4 Hz, 2H), 7.31 (d,J = 8.4 Hz, 2H), 4.58 (t, J = 5.2 Hz, 1H), 4.01 (d, J = 6.0 Hz, 2H), 2.43 (s,3H), 1.29 (s, 9H) .

[0223] Step 8: Synthesis of N-(1-(((tert-butyldiphenylsilyl)oxy)but-3-yn-2-yl)-N-(3-(tert-butylthio)prop-2-yn-1-yl)-4-methylbenzenesulfonamide

[0224] N-(3-(tert-butylthio)prop-2-yn-1-yl)-4-methylbenzenesulfonamide (2.3 g, 7.73 mmol), 1-((tert-butyldiphenylsilyl)oxy)but-3-yn-2-ol (2.5 g, 7.73 mmol), and triphenylphosphine (2.4 g, 9.28 mmol) were dissolved in tetrahydrofuran (20 mL), purged three times with nitrogen, and diisopropyl azodicarbonate (1.8 mL, 9.28 mmol) was slowly added at 0 °C. The reaction mixture was stirred at 20 °C for 1 hour. The organic phase was poured into water (60 mL), and then extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, and filtered to obtain the crude product. The crude product was purified by normal-phase silica gel column chromatography and then purified by reverse-phase chromatography (formic acid system) to obtain 2.6 g of the product.

[0225] LC-MS (ESI): [M+H] + =604.2;

[0226] Step 9: Synthesis of 4-(((tert-butyldiphenylsilyl)oxy)methyl)-5-p-tolyl-5,6-dihydro-4H-thieno[3,4-c]pyrrole

[0227] N-(1-(((tert-butyldiphenylsilyl)oxy)but-3-yn-2-yl)-N-(3-(tert-butylthio)prop-2-yn-1-yl)-4-methylbenzenesulfonamide (2.5 g, 4.14 mmol) was dissolved in acetic acid (50 mL), and silver acetate (69.1 mg, 0.41 mmol) was added. The reaction mixture was stirred at 20 °C for 1 hour. The reaction mixture was directly concentrated under reduced pressure and evaporated to dryness to obtain the crude product. The crude product was purified by normal-phase silica gel column chromatography to obtain 2.1 g of the product.

[0228] LC-MS (ESI): [M+H] + =548.3;

[0229] Step 10: Synthesis of (5-toluenesulfonyl-5,6-dihydro-4H-thieno[3,4-c]pyrrolo-4-yl)methanol

[0230] 4-(((tert-butyldiphenylsilyl)oxy)methyl)-5-p-tolyl-5,6-dihydro-4H-thieno[3,4-c]pyrrole (3.7 mL, 3.65 mmol) was dissolved in tetrahydrofuran (20 mL), and a tetrabutylammonium fluoride solution (11.0 mL, 10.95 mmol) in tetrahydrofuran was added. The reaction mixture was stirred at 25 °C for 2 hours. The reaction mixture was directly extracted with water (50 mL) and ethyl acetate (50 mL × 2). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated to obtain crude product, which was purified by normal-phase silica gel column chromatography to give 905 mg of product.

[0231] LC-MS (ESI): [M+H] + =310.1;

[0232] Step 11: Synthesis of 5-toluenesulfonyl-5,6-dihydro-4H-thieno[3,4-c]pyrrole-4-carboxaldehyde

[0233] (5-Toluenesulfonyl-5,6-dihydro-4H-thieno[3,4-c]pyrrolo-4-yl)methanol (0.75 g, 2.42 mmol) was dissolved in acetonitrile (20 mL), and 2-iodobenzoic acid (5.43 g, 19.39 mmol) was added. The reaction solution was reacted at 60 °C for 16 hours. The reaction solution was directly filtered, and the filter cake was washed with acetonitrile (30 mL × 2). The organic phases were combined and directly concentrated to give 700 mg of product.

[0234] LC-MS (ESI): [M+H] + =308.0;

[0235] Step 12: Synthesis of 5-toluenesulfonyl-5,6-dihydro-4H-thieno[3,4-c]pyrrole-4-carboxylic acid

[0236] 5-Toluenesulfonyl-5,6-dihydro-4H-thieno[3,4-c]pyrrole-4-carboxaldehyde (0.70 g, 2.28 mmol) and 2-methylisobutylene (2.4 mL, 22.77 mmol) were dissolved in tert-butanol (5 mL), and a prepared solution of sodium chlorite (0.82 g, 9.11 mmol) and sodium dihydrogen phosphate (488 mg, 0.65 mmol) in water (2 mL) was added. The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was extracted with saturated brine (50 mL) and ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude product obtained by direct concentration of the filtrate was purified by normal-phase silica gel column chromatography to give 430 mg of the product.

[0237] LC-MS (ESI): [M+H] + =324.1;

[0238] Step 13: Synthesis of methyl 4'-(5-toluenesulfonyl-5,6-dihydro-4H-thieno[3,4-c]pyrrole-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0239] 5-Toluenesulfonyl-5,6-dihydro-4H-thieno[3,4-c]pyrrole-4-carboxylic acid (400 mg, 1.24 mmol) and methyl 4-(4-aminophenyl)benzoate (281.11 mg, 1.24 mmol) were dissolved in dichloromethane (5 mL), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (308.26 mg, 1.61 mmol) was added. The reaction mixture was reacted at 25°C for 16 hours. The reaction mixture was extracted with aqueous solution (20 mL) and dichloromethane (20 mL × 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated to obtain crude product, which was purified by normal-phase silica gel column chromatography to give 460 mg of product.

[0240] LC-MS (ESI): [M+H] + =533.0;

[0241] Step 14: Synthesis of 4'-(5,6-dihydro-4H-thieno[3,4-c]pyrrole-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0242] Methyl 4'-(5-toluenesulfonyl-5,6-dihydro-4H-thieno[3,4-c]pyrrolo-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (50 mg, 0.09 mmol) and phenol (0.1 mL, 0.94 mmol) were dissolved in hydrobromic acid and 33% w / w (45% w / v) acetic acid solution (1 mL). The reaction mixture was heated to 50 °C and reacted for 16 hours. Water (1 mL) and acetonitrile (1 mL) were added to the reaction mixture. A solid precipitated at this point, and the reaction mixture was directly filtered. The filter cake was collected to give 12 mg of product.

[0243] LC-MS (ESI): [M+H] + =365.1;

[0244] Step 15: Synthesis of 4'-(5-(((4-isopropylphenyl)carbamoyl)-5,6-dihydro-4H-thieno[3,4-c]pyrrolo-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0245] 4'-(5,6-dihydro-4H-thieno[3,4-c]pyrrolo-4-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (12 mg, 0.03 mmol) was dissolved in tetrahydrofuran (0.5 mL), and triethylamine (10 mg, 0.10 mmol) and 4-isocyano-1-(prop-2-yl)benzene (7 mg, 0.04 mmol) were added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was quenched with methanol (10 mL). The crude product obtained by direct concentration of the reaction mixture was purified by high performance liquid chromatography, and the fraction was lyophilized to obtain 5 mg of product.

[0246] LC-MS (ESI): [M+H] + =526.2;

[0247] 1 H NMR (400 MHz, DMSO-d6) δ = 10.58 - 10.42 (m, 1H), 8.45 - 8.39 (m,1H), 7.97 (d, J = 8.4 Hz, 2H), 7.76 - 7.67 (m, 6H), 7.48 - 7.39 (m, 3H), 7.38- 7.34 (m, 1H), 7.12 (d, J = 8.6 Hz, 2H), 5.55 - 5.45 (m, 1H), 4.81 - 4.64(m, 2H), 2.87 - 2.77 (m, 1H), 1.17 (d, J = 6.8 Hz, 6H).

[0248] Example 8: (R)-4'-(2-(((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid, (S)-4'-(2-(((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid (compounds 8A and 8B)

[0249]

[0250] Step 1: Synthesis of tert-butyl 1-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-2-azaspiro[3,4]octane-2-carboxylic acid

[0251] The starting material 2-(tert-butoxycarbonyl)-2-azaspiro[3,4]octane-1-carboxylic acid (105 mg, 0.41 mmol) was dissolved in dichloromethane (5 mL), and methyl 4'-amino-[1,1'-biphenyl]-4-carboxylic acid (105 mg, 0.46 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (105 mg, 0.55 mmol) were added separately. The reaction mixture was stirred at 25 °C for 12 hours. The mixture was diluted with water (50 mL) and ethyl acetate (50 mL), separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated to remove the solvent. The crude product was purified by normal-phase column chromatography, and the fraction was concentrated to remove the solvent, yielding 190 mg of product.

[0252] LC-MS (ESI) m / z: [M+H] + =465.3;

[0253] Step 2: Synthesis of 4'-(2-(tert-butoxycarbonyl)-2-azaspiro[3,4]octane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0254] The starting material 1-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-2-azaspiro[3,4]octane-2-carboxylic acid tert-butyl ester (190 mg, 0.41 mmol) was dissolved in acetonitrile (6 mL), and potassium trimethylsilanolate (315 mg, 2.45 mmol) was added. The reaction mixture was stirred at 25 °C for 3 hours. The mixture was diluted with water (50 mL) and ethyl acetate (50 mL), and the pH was adjusted to 3-4 with 1 molar hydrochloric acid solution. The mixture was separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to remove the solvent, yielding 190 mg of the product.

[0255] LC-MS (ESI) m / z: [M+H] + =451.1;

[0256] Step 3: Synthesis of 4'-(2-azaspiro[3.4]octane-1-formamido)-[1,1'-biphenyl]-4-carboxylic acid hydrochloride

[0257] The starting material 4'-(2-(tert-butoxycarbonyl)-2-azaspiro[3.4]octane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (180 mg, 0.40 mmol) was dissolved in 4 mol of dioxane hydrochloride solution (10 mL). The reaction solution was stirred at 25 °C for 1 hour. The reaction solution was diluted with dichloromethane and concentrated to remove the solvent, yielding 140 mg of product.

[0258] LC-MS (ESI) m / z: [M+H] + =351.2;

[0259] Step 4: Synthesis of 4'-(2-(((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0260] The starting material 4'-(2-azaspiro[3,4]octane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid hydrochloride (120 mg, 0.31 mmol) was dissolved in tetrahydrofuran (5 mL), and triethylamine (174 mg, 1.71 mmol) was added. The reaction mixture was stirred at 25 °C for 3 minutes, and then 2-fluoro-4-isocyanate-1-isopropylbenzene (123 mg, 0.68 mmol) was added. The reaction mixture was stirred at 25 °C for 12 hours. The reaction mixture was quenched with methanol, and the solvent was removed by concentration to obtain the crude product. The crude product was separated by high performance liquid chromatography, and the fraction was lyophilized to obtain 135 mg of the product.

[0261] LC-MS (ESI) m / z: [M+H] + =530.3;

[0262] Step 5: Synthesis of (R)-4'-(2-(((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid and (S)-4'-(2-(((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid

[0263] The starting material 4'-(2-(((3-fluoro-4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid (130 mg, 0.25 mmol) was purified by SFC to obtain compound 8A: 40 mg, retention time 1.175 min; and compound 8B: 40 mg, retention time 1.515 min.

[0264] Compound 8A:

[0265] LC-MS (ESI): [M+H] + =530.1;

[0266] 1H NMR (400 MHz, DMSO- d6) δ = 10.05 (s, 1H), 8.66 (s, 1H), 7.99 (d,J = 8.4 Hz, 2H), 7.79 - 7.76 (m, 4H), 7.72 (d, J = 8.8 Hz, 2H), 7.41 (dd, J =2.0, 13.6 Hz, 1H), 7.21 - 7.15 (m, 2H), 4.57 (s, 1H), 3.91 (d, J = 7.2 Hz,1H), 3.80 (d, J = 7.2 Hz, 1H), 3.08 - 3.07 (m, 1H), 1.95 - 1.91 (m, 2H), 1.64- 1.52 (m, 6H), 1.17 (d, J = 6.8 Hz, 6H).

[0267] Compound 8B:

[0268] LC-MS (ESI): [M+H] + =530.1;

[0269] 1 H NMR (400 MHz, DMSO-d6) δ = 10.07 (s, 1H), 8.67 (s, 1H), 7.99 (d, J= 8.4 Hz, 2H), 7.79 - 7.76 (m, 4H), 7.72 (d, J = 8.8 Hz, 2H), 7.42 (dd, 1.95 - 1.93 (m, 2H), 1.64- 1.52 (m, 6H), 1.17 (d, J = 7.2 Hz, 6H).

[0270] Example 9: (R)-4'-(2-(((4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid, (S)-4'-(2-((4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid (compounds 9A and 9B)

[0271]

[0272] Step 1: Synthesis of tert-butyl 1-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-2-azaspiro[3,4]octane-2-carboxylic acid

[0273] The starting materials 2-(tert-butoxycarbonyl)-2-azaspiro[3.4]octane-1-carboxylic acid (100 mg, 0.39 mmol) and methyl 4'-amino-[1,1'-biphenyl]-4-carboxylic acid (115.72 mg, 0.51 mmol) were dissolved in dichloromethane (3 mL), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (120.13 mg, 0.63 mmol) was added. The reaction mixture was stirred at 20 °C for 2 hours. The reaction mixture was extracted with water (20 mL) and dichloromethane (20 mL × 3), and the liquid was separated. The organic phase was washed once with saturated sodium chloride aqueous solution (20 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to remove the solvent, and the crude product was purified by normal-phase column chromatography. The fraction was concentrated to remove the solvent to obtain 183 mg of product.

[0274] LC-MS (ESI) m / z: [M+Na] + =487.3;

[0275] Step 2: Synthesis of 4'-(2-(tert-butoxycarbonyl)-2-azaspiro[3,4]octane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0276] The starting material 1-((4'-(methoxycarbonyl)-[1,1'-biphenyl]-4-yl)carbamoyl)-2-azaspiro[3,4]octane-2-carboxylic acid tert-butyl ester (180 mg, 0.39 mmol) was dissolved in acetonitrile (8 mL), and potassium trimethylsilanolate (298.24 mg, 2.32 mmol) was added. The reaction mixture was stirred at 20 °C for 12 hours. Water (20 mL) was added to the reaction mixture, and the pH of the aqueous phase was adjusted to 5-6 with 1 mol hydrochloric acid solution, followed by extraction with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated under reduced pressure to remove the solvent, yielding 170 mg of the product.

[0277] LC-MS (ESI) m / z:[M+H] + =451.3;

[0278] Step 3: Synthesis of 4'-(2-azaspiro[3.4]octane-1-formamido)-[1,1'-biphenyl]-4-carboxylic acid trifluoroacetate

[0279] The starting material 4'-(2-(tert-butoxycarbonyl)-2-azaspiro[3.4]octane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (170 mg, 0.38 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (2 mL, 26.93 mmol) was added. The reaction mixture was stirred at 20 °C for 1 hour. The reaction mixture was then directly concentrated under reduced pressure to remove the solvent, yielding 132 mg of the product.

[0280] Step 4: Synthesis of 4'-(2-(((4-isopropylphenyl)carbamoyl)-2-azaspiro[3,4]octane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0281] The starting material 4'-(2-azaspiro[3,4]octane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid trifluoroacetate (130 mg, 0.28 mmol) was dissolved in tetrahydrofuran (4 mL), and triethylamine (225.24 mg, 2.23 mmol) was added. The reaction mixture was stirred at 25°C for 3 minutes, and then 1-isocyanate-4-isopropylbenzene (119.61 mg, 0.74 mmol) was added. The reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched with methanol, and the solvent was removed by concentration. The crude product was separated by high performance liquid chromatography, and the fraction was lyophilized to give 122 mg of the product.

[0282] LC-MS (ESI) m / z:[M+H] + =512.2;

[0283] Step 5: Synthesis of (R)-4'-(2-(((4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid and (S)-4'-(2-((4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid

[0284] The starting material 4'-(2-(((4-isopropylphenyl)carbamoyl)-2-azaspiro[3.4]octane-1-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (120 mg, 0.23 mmol) was separated by supercritical fluid chromatography (SFC). Fraction 1 was concentrated to remove the solvent, yielding compound 9A: 51.01 mg, retention time 1.590 min; Fraction 2 was concentrated to remove the solvent, yielding compound 9B: 40.21 mg, retention time 1.311 min.

[0285] Compound 9A: LC-MS (ESI): [M+H] + =512.2;

[0286] 1 H NMR (400 MHz, DMSO-d6) δ = 10.04 (s, 1H), 8.46 (s, 1H), 7.99 (d, J= 8.4 Hz, 2H), 7.84 - 7.68 (m, 6H), 7.41 (d, J = 8.4 Hz, 2H), 7.10 (d, J =8.4 Hz, 2H), 4.56 (s, 1H), 3.90 (d, J = 7.2 Hz, 1H), 3.79 (d, J = 7.2 Hz, 1H), 2.85 - 2.73 (m, 1H), 2.04 - 1.83 (m, 2H), 1.74 - 1.47 (m, 6H), 1.16 (d,J = 6.8 Hz, 6H).

[0287] Compound 9B: LC-MS (ESI): [M+H] + =512.2;

[0288] 1 H NMR (400 MHz, DMSO-d6) δ = 10.05 (s, 1H), 8.47 (s, 1H), 7.99 (d, J= 8.4 Hz, 2H), 7.83 - 7.66 (m, 6H), 7.41 (d, J = 8.4 Hz, 2H), 7.10 (d, J =8.4 Hz, 2H), 4.56 (s, 1H), 3.90 (d, J = 7.2 Hz, 1H), 3.79 (d, J = 7.2 Hz, 1H), 2.85 - 2.75 (m, 1H), 2.01 - 1.85 (m, 2H), 1.75 - 1.46 (m, 6H), 1.16 (d,J = 6.8 Hz, 6H).

[0289] Example 10: 4'-(((1S,3R,4S)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid, 4'-((1R,3S,4R)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid (compounds 10A and 10B)

[0290]

[0291] Step 1: Synthesis of 4'-(((1S,4S)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid

[0292] The starting material 4'-(((1S,4S)-2-azabicyclo[2.1.1]hexane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid trifluoroacetate (90 mg, 0.21 mmol) was dissolved in tetrahydrofuran (4 mL), and triethylamine (108 mg, 1.06 mmol) was added. The mixture was stirred for 3 minutes. Then, 1-isocyanate-4-isopropylbenzene (69 mg, 0.42 mmol) was added. The reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched with methanol, concentrated to remove the solvent, and the crude product was separated by high performance liquid chromatography. The fraction was lyophilized to obtain 60 mg of the product.

[0293] LC-MS (ESI) m / z:[M+H] + =484.3;

[0294] Step 2: Synthesis of 4'-(((1S,3R,4S)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid and 4'-((1R,3S,4R)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)-[1,1'-biphenyl]-4-carboxylic acid

[0295] The starting material 4'-(((1S,4S)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carboxamido)-[1,1'-biphenyl]-4-carboxylic acid (50 mg, 0.10 mmol) was separated by supercritical fluid chromatography. Fraction 1 was concentrated to remove the solvent to obtain compound 10A: 22 mg, retention time 1.315 min; fraction 2 was concentrated to remove the solvent to obtain compound 10B: 20 ​​mg, retention time 1.625 min.

[0296] Compound 10A: LC-MS (ESI): [M+H] + =484.2;

[0297] 1¹H NMR (400 MHz, DMSO-d6) δ = 10.21 (s, 1H), 8.59 (s, 1H), 7.99 (d, J= 8.4 Hz, 2H), 7.77 (d, J = 8.4 Hz, 4H), 7.72 (d, J = 8.8 Hz, 2H), 7.43 (d, J= 8.4 Hz, 2H), 7.12 (d, J = 8.8 Hz, 2H), 4.74 (d, J = 6.4 Hz, 1H), 4.43 (s, 1H), 3.05 - 3.03 (m, 1H), 2.83 - 2.80 (m, 1H), 2.10 - 2.09 (m, 1H), 1.97 - 1.95 (m, 1H), 1.83 - 1.81 (m, 1H), 1.43 - 1.41 (m, 1H), 1.17 (d, J = 7.2 Hz, 6H).

[0298] Compound 10B: LC-MS (ESI): [M+H] + = 484.2;

[0299] 1 ¹H NMR (400 MHz, DMSO-d6) δ = 10.21 (s, 1H), 8.59 (s, 1H), 7.98 (d, J= 8.4 Hz, 2H), 7.77 - 7.75 (m, 4H), 7.71 (d, J = 9.2 Hz, 2H), 7.43 (d, J = 8.4 Hz, 2H), 7.12 (d, J = 8.4 Hz, 2H), 4.73 (d, J = 6.8 Hz, 1H), 4.43 (s, 1H), 3.05 - 3.03 (m, 1H), 2.84 - 2.80 (m, 1H), 2.10 - 2.08 (m, 1H), 1.97 - 1.94 (m, 1H), 1.83 - 1.81 (m, 1H), 1.43 - 1.38 (m, 1H), 1.17 (d, J = 7.2 Hz, 6H).

[0300] Example 11: 5-(4-(((1S,3R,4S)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)phenyl)-6-methylpyridinecarboxylic acid, 5-(4-((1R,3S,4R)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)phenyl)-6-methylpyridinecarboxylic acid (compounds 11A and 11B)

[0301]

[0302] Step 1: Synthesis of (1S,4S)-3-((4-(6-(methoxycarbonyl)-2-methylpyridin-3-yl)phenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-2-carboxylic acid tert-butyl ester

[0303] (1S,4S)-3-{[(2-methylprop-2-yl)oxy]carbonyl}-3-azabicyclo[2.1.1]hexane-2-carboxylic acid (80 mg, 0.31 mmol) and methyl 5-(4-aminophenyl)-6-methylpyridine-2-carboxylate (82.09 mg, 0.34 mmol) were dissolved in dichloromethane (1 mL), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (88.57 mg, 0.46 mmol) was added. The reaction mixture was reacted at 25 °C for 1 hour. The reaction mixture was extracted with water (30 mL) and dichloromethane (20 mL × 3). The combined organic phases were washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The crude product obtained by direct concentration of the filtrate was purified by normal-phase silica gel column chromatography to give 131 mg of product.

[0304] LC-MS (ESI) m / z:[M+H] + =452.3;

[0305] Step 2: Synthesis of 5-(4-(((1S,4S)-2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-3-carboxamido)phenyl)-6-methylpyridinecarboxylic acid

[0306] (1S,4S)-3-((4-(6-(methoxycarbonyl)-2-methylpyridin-3-yl)phenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-2-carboxylic acid tert-butyl ester (131 mg, 0.29 mmol) was dissolved in acetonitrile (5 mL), and potassium trimethylsilanolate (186.10 mg, 1.45 mmol) was added. The reaction solution was reacted at 25 °C for 2 hours. Water (10 mL) was added to the reaction solution, and the pH of the aqueous phase was adjusted to 5-6 with 1 mol hydrochloric acid aqueous solution. The aqueous phase was extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated under reduced pressure to give 125 mg of product.

[0307] LC-MS (ESI) m / z:[M+H] + =438.1;

[0308] Step 3: Synthesis of 5-(4-(((1S,4S)-2-azabicyclo[2.1.1]hexane-3-carboxamido)phenyl)-6-methylpyridinecarboxylic acid trifluoroacetate

[0309] 5-(4-(((1S,4S)-2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-3-carboxamido)phenyl)-6-methylpyridinecarboxylic acid (125 mg, 0.29 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (1.1 mL, 14.29 mmol) was added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove the solvent, yielding 121 mg of the product.

[0310] LC-MS (ESI) m / z:[M+H] + =338.2;

[0311] Step 4: Synthesis of 5-(4-(((1S,4S)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)phenyl)-6-methylpyridinecarboxylic acid

[0312] 5-(4-(((1S,4S)-2-azabicyclo[2.1.1]hexane-3-carboxamido)phenyl)-6-methylpyridinecarboxylic acid trifluoroacetate (121 mg, 0.27 mmol) was dissolved in tetrahydrofuran (3 mL), and triethylamine (0.2 mL, 1.34 mmol) and 4-isocyano-1-(prop-2-yl)benzene (0.1 mL, 0.54 mmol) were added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was quenched with methanol (10 mL), and then concentrated under reduced pressure to remove the solvent to obtain the crude product. The crude product was purified by high performance liquid chromatography, and the fraction was lyophilized to obtain 60 mg of the product.

[0313] LC-MS (ESI) m / z:[M+H] + =499.1;

[0314] Step 5: Synthesis of 5-(4-(((1S,3R,4S)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)phenyl)-6-methylpyridinecarboxylic acid and 5-(4-((1R,3S,4R)-2-((4-isopropylphenyl)carbamoyl)-2-azabicyclo[2.1.1]hexane-3-carbamoyl)phenyl)-6-methylpyridinecarboxylic acid

[0315] Racemic 6-methyl-5-[4-({[(1S,4S)-3-({[4-(propyl-2-yl)phenyl]amino}carbonyl)-3-azabicyclo[2.1.1]hex-2-yl]carbonyl}amino)phenyl]pyridine-2-carboxylic acid (60 mg, 0.12 mmol) was purified by supercritical fluid chromatography with chiral resolution. Fraction 1 was lyophilized to give product 11A: 13 mg, retention time 1.775 min; fraction 2 was lyophilized to give product 11B: 15 mg, retention time 1.126 min.

[0316] Compound 11A: LC-MS (ESI): [M+H] + =499.4;

[0317] 1¹H NMR (400 MHz, DMSO-d6) δ = 10.24 (s, 1H), 8.60 (s, 1H), 7.93 - 7.87(m, 1H), 7.80 - 7.71 (m, 3H), 7.47 - 7.38 (m, 4H), 7.16 – 7.09 (m, 2H), 4.77- 4.70 (m, 1H), 4.44 (s, 1H), 3.07 - 3.04 (m, 1H), 2.86 - 2.80 (m, 1H), 2.50- 2.49 (m, 3H), 2.09 (br s, 1H), 1.99 - 1.94 (m, 1H), 1.86 - 1.80 (m, 1H),1.43 (br s, 1H), 1.18 (d, J = 6.8 Hz, 6H).

[0318] Compound 11B: LC-MS (ESI): [M+H] + = 499.4;

[0319] 1 ¹H NMR (400 MHz, DMSO-d6) δ = 10.24 (s, 1H), 8.60 (s, 1H), 7.91 - 7.83(m, 1H), 7.79 - 7.74 (m, 2H), 7.73 - 7.67 (m, 1H), 7.46 - 7.41 (m, 2H), 7.41- 7.36 (m, 2H), 7.12 (d, J = 8.8 Hz, 2H), 4.76 - 4.71 (m, 1H), 4.44 (s, 1H),3.05 - 3.02 (m, 1H), 2.86 - 2.78 (m, 1H), 2.48 (br s, 3H), 2.13 - 2.06 (m,1H), 1.98 - 1.93 (m, 1H), 1.85 - 1.79 (m, 1H), 1.44 - 1.33 (m, 1H), 1.17 (d,J = 6.8 Hz, 6H).

[0320] Example 12: 4-[4-({[(1S,4S)-2-({[4-(prop-2-yl)phenyl]amino}carbonyl)-2-aza-6-oxaspiro[3.4]oct-1-yl]carbonyl}amino)phenyl]benzoic acid, 4-[4-({[(1R,4S)-2-({[4-(prop-2-yl)phenyl]amino}carbonyl)-2-aza-6-oxaspiro[3.4]oct-1-yl]carbonyl}amino)phenyl]benzoic acid, 4-[4-({[(1S,4R)-2-({[4-(prop-2-yl)phenyl]amino}carbonyl)-2-aza-6-oxaspiro[3.4]oct-1-yl]carbonyl}amino)phenyl]benzoic acid, 4-[4-({[(1R,4R)-2-({[4-(prop-2-yl)phenyl]amino}carbonyl)-2-aza-6-oxaspiro[3.4]oct-1-yl]carbonyl}amino)phenyl]benzoic acid (compounds 12A, 12B, 12C, and 12D)

[0321]

[0322] Step 1: Synthesis of tert-butyl 1-{[(4-{4-[(benzyloxy)carbonyl]phenyl}phenyl)amino]carbonyl}-2-aza-6-oxaspiro[3,4]octane-2-carboxylate

[0323] 2-{[(2-methylprop-2-yl)oxy]carbonyl}-2-aza-5-oxaspiro[3,4]octane-1-carboxylic acid (100 mg, 0.39 mmol) and benzyl 4-(4-aminophenyl)benzoate (82 mg, 0.27 mmol) were dissolved in dichloromethane (4 mL), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (112 mg, 0.29 mmol) and diisopropylethylamine (0.10 mL, 0.61 mmol) were added. The mixture was reacted at room temperature for 4 hours, and the product was purified by column chromatography after concentration to obtain 151 mg of the product.

[0324] LC-MS(ESI): [M+H] + =543.4;

[0325] Step 2: Synthesis of 4-{4-[(2-aza-6-oxaspiro[3.4]oct-1-ylcarbonyl)amino]phenyl}benzyl benzoate hydrochloride

[0326] 151 mg, 0.18 mmol of tert-butyl 1-{[(4-{4-[(benzyloxy)carbonyl]phenyl}phenyl)amino]carbonyl}-2-aza-6-oxaspiro[3,4]octane-2-carboxylate was added to a solution of 1,4-dioxane (3 mL, 4N) in hydrochloric acid, and the reaction was carried out at room temperature for 1 hour. The solvent was removed by direct concentration to give 91.00 mg of the product.

[0327] Step 3: Synthesis of benzyl benzoate 4-[4-({[2-({[4-(propyl-2-yl)phenyl]amino}carbonyl)-2-aza-6-oxaspiro[3,4]oct-1-yl]carbonyl}amino)phenyl]benzoate

[0328] 4-(prop-2-yl)aniline (0.052 mL, 0.38 mmol) was dissolved in dichloromethane (2 mL), and triphosgene (38 mg, 0.13 mmol) was added under ice-water bath conditions. The reaction was carried out at this temperature for 2 hours.

[0329] 4-{4-[(2-aza-6-oxaspiro[3.4]oct-1-ylcarbonyl)amino]phenyl}benzyl benzoate hydrochloride (91.0 mg, 0.19 mmol) was dissolved in dichloromethane (2 mL), followed by the addition of triethylamine (0.11 mL, 0.76 mmol), and then freshly prepared isocyanate solution was added dropwise. The reaction was allowed to proceed overnight at room temperature. After concentration, the product was purified by column chromatography to give 88 mg of the product.

[0330] LC-MS(ESI): [M+H] + =604.6;

[0331] Step 4: 4-[4-({[(1S,4S)-2-({[4-(prop-2-yl)phenyl]amino}carbonyl)-2-aza-6-oxaspiro[3.4]oct-1-yl]carbonyl}amino)phenyl]benzoic acid, 4-[4-({[(1R,4S)-2-({[4-(prop-2-yl)phenyl]amino}carbonyl)-2-aza-6-oxaspiro[3.4]oct-1-yl]carbonyl}amino)phenyl]benzoic acid, Synthesis of 4-[4-({[(1S,4R)-2-({[4-(prop-2-yl)phenyl]amino}carbonyl)-2-aza-6-oxaspiro[3.4]oct-1-yl]carbonyl}amino)phenyl]benzoic acid and 4-[4-({[(1R,4R)-2-({[4-(prop-2-yl)phenyl]amino}carbonyl)-2-aza-6-oxaspiro[3.4]oct-1-yl]carbonyl}amino)phenyl]benzoic acid

[0332] Benzyl benzoate (88 mg, 0.15 mmol) was dissolved in tetrahydrofuran (3 mL), and 5% palladium / carbon (20 mg, mmol) was added. The mixture was hydrogenated overnight at room temperature. After filtration and concentration, the product was separated by SFC. Fraction 1 was lyophilized to give compound 12A: 2 mg, retention time 1.781 min; fraction 2 was lyophilized to give compound 12B: 3 mg, retention time 1.544 min; fraction 3 was lyophilized to give compound 12C: 5 mg, retention time 1.617 min; and fraction 4 was lyophilized to give compound 12D: 3 mg, retention time 1.293 min.

[0333] Compound 12A: LC-MS (ESI): [M+H] + =514.3;

[0334] 1 H NMR (400 MHz, DMSO-d6) δ10.26 (s, 1H), 8.63 (s, 1H), 8.03 (d, J =8.1 Hz, 2H), 7.81 (d, J = 8.7 Hz, 2H), 7.76 (d, J = 8.9 Hz, 4H), 7.46 (d, J =8.5 Hz, 2H), 7.17 (d, J = 8.6 Hz, 2H), 4.80 (s, 1H), 4.11 (d, J = 7.7 Hz,1H), 4.03 (d, J = 7.5 Hz, 1H), 3.86 (dd, J = 14.3, 7.8 Hz, 1H), 3.81 – 3.77(m, 1H), 3.75 – 3.70 (m, 2H), 2.91 – 2.82 (m, 1H), 2.41 – 2.28 (m, 2H), 1.22(d, J = 6.9 Hz, 6H).

[0335] Compound 12B: LC-MS (ESI): [M+H] + =514.3;

[0336] 1¹H NMR (400 MHz, DMSO-d6) δ10.20 (s, 1H), 8.58 (s, 1H), 7.95 (d, J =8.2 Hz, 2H), 7.77 (d, J = 8.7 Hz, 2H), 7.72 – 7.61 (m, 4H), 7.41 (d, J = 8.5Hz, 2H), 7.11 (d, J = 8.5 Hz, 2H), 4.69 (s, 1H), 4.04 (d, J = 7.5 Hz, 1H),3.99 – 3.90 (m, 3H), 3.74 – 3.66 (m, 2H), 2.85 – 2.76 (m, 1H), 2.00 – 1.92(m, 2H), 1.17 (d, J = 6.9 Hz, 6H).

[0337] Compound 12C: LC-MS(ESI): [M+H] + =514.3;

[0338] 1 ¹H NMR (400 MHz, DMSO-d6) δ10.24 (s, 1H), 8.62 (s, 1H), 8.05 (d, J =8.2 Hz, 2H), 7.88 – 7.73 (m, 6H), 7.47 (d, J = 8.4 Hz, 2H), 7.17 (d, J = 8.5Hz, 2H), 4.74 (s, 1H), 4.10 (d, J = 7.4 Hz, 1H), 4.05 – 3.97 (m, 3H), 3.78 –3.72 (m, 2H), 2.87 (dt, J = 13.7, 6.9 Hz, 1H), 2.06 – 1.97 (m, 2H), 1.23 (d,J = 6.9 Hz, 6H).

[0339] Compound 12D: LC-MS(ESI): [M+H] + =514.3;

[0340] 1H NMR (400 MHz, DMSO-d6) δ10.21 (s, 1H), 8.57 (s, 1H), 7.99 (d, J =8.3 Hz, 2H), 7.81 – 7.68 (m, 6H), 7.41 (d, J = 8.5 Hz, 2H), 7.11 (d, J = 8.6Hz, 2H), 4.74 (s, 1H), 4.06 (d, J = 7.6 Hz, 1H), 3.97 (d, J = 7.7 Hz, 1H), 3.81 (dd, J = 14.4, 7.6 Hz, 1H), 3.76 – 3.71 (m, 1H), 3.71 – 3.63 (m, 2H),2.81 (dt, J = 13.7, 6.8 Hz, 1H), 2.35 – 2.22 (m, 2H), 1.17 (d, J = 6.9 Hz, 6H).

[0341] Example 13: 4-(4-((((3R)-2-(((4-(prop-2-yl)phenyl)amino)carbonyl)tetrahydroisoxazole-3-yl)carbonyl)amino)phenyl)benzoic acid, 4-(4-((((3S)-2-(((4-(prop-2-yl)phenyl)amino)carbonyl)tetrahydroisoxazole-3-yl)carbonyl)amino)phenyl)benzoic acid (compounds 13A and 13B)

[0342]

[0343] Step 1: Synthesis of (((dimethyl(tert-butyl)silyl)oxy)amino)methane-tert-butyl ester)

[0344] Dissolve ((tert-butyl)oxy)methanehydroxyline (10 g, 75.10 mmol) and triethylamine (11.5 mL, 82.62 mmol) in dichloromethane (100 mL). After cooling to 0°C, add dropwise a solution of tert-butyldimethylchlorosilane (13.0 mL, 75.10 mmol) in dichloromethane (100 mL). React the solution at 25°C for 16 hours. Add water (100 mL) to the reaction solution, separate the layers, and extract the aqueous phase once with dichloromethane (100 mL). Combine the organic phases, wash with saturated brine (100 mL), dry to anhydrous sodium sulfate, and filter. Concentrate the filtrate directly to remove the solvent, yielding 18 g of product.

[0345] 1H NMR (400 MHz, CHLOROFORM-d) δ = 6.70 (s, 1H), 1.48 (s, 9H), 0.96 (s, 9H), 0.21 - 0.15 (m, 6H).

[0346] Step 2: Synthesis of ethyl 3-formyl-2-(2,2,7,7,8,8-hexamethyl-4-oxoylide-5-aza-3,6-dioxa-7-silylen-5-yl)propionate

[0347] Ethyl (2E)-3-formylpropionic-2-enoate (7.2 g, 56.19 mmol) was dissolved in dichloromethane (40 mL). After cooling to 0 °C, tetrahydropyrrole (2.3 mL, 28.10 mmol) was added, and the mixture was stirred for 10 minutes. Then, a solution of (((dimethyl(tert-butyl)silyl)oxy)amino)methane-tert-butyl ester (16.68 g, 67.43 mmol) in dichloromethane (40 mL) was added dropwise. The reaction mixture was directly concentrated to remove the solvent, and the crude product was purified by normal-phase silica gel column chromatography to obtain 3.5 g of the product.

[0348] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 9.84 (s, 1H), 4.93 (dd, J = 5.7,7.6 Hz, 1H), 4.24 - 4.18 (m, 2H), 3.25 (dd, J = 8.1, 17.3 Hz, 1H), 2.81 (dd,J = 4.8, 17.7 Hz, 1H), 1.49 (s, 9H), 1.25 (br d, J = 6.0 Hz, 3H), 0.93 (s,9H), 0.17 (s, 6H).

[0349] Step 3: Synthesis of ethyl 2-(2,2,7,7,8,8-hexamethyl-4-oxoylide-5-aza-3,6-dioxa-7-silylen-5-yl)-4-hydroxybutyrate

[0350] Ethyl 3-formyl-2-(2,2,7,7,8,8-hexamethyl-4-oxoylide-5-aza-3,6-dioxa-7-silanon-5-yl)propionate (3500 mg, 9.32 mmol) was dissolved in methanol (35 mL), cooled to -20 °C, and sodium borohydride (706 mg, 18.64 mmol) was added in portions. After the addition was complete, the reaction mixture was heated to 0 °C and reacted for 1 hour. Water (20 mL) was added to the reaction mixture, and after stirring for 2 minutes, ethyl acetate (20 mL × 3) was added for extraction. The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated to obtain a crude product, which was purified by normal-phase silica gel column chromatography to yield 1.3 g of the product.

[0351] Step 4: Synthesis of 6-(ethoxycarbonyl)-2,2,3,3-tetramethyl-5-(((tert-butyl)oxy)carbonyl)-5-aza-4-oxa-3-silazaoct-8-yl methanesulfonate

[0352] Ethyl 2-(2,2,7,7,8,8-hexamethyl-4-oxoylide-5-aza-3,6-dioxa-7-silanenon-5-yl)-4-hydroxybutyrate (1.30 g, 3.44 mmol) and triethylamine (1.4 mL, 10.33 mmol) were dissolved in dichloromethane (13 mL). After cooling to 0 °C, methanesulfonyl chloride (0.5 mL, 6.98 mmol) was added, and the reaction was carried out at room temperature for 1 hour. The reaction solution was directly extracted with water (20 mL) and dichloromethane (20 mL × 2). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated to give 1.35 g of product.

[0353] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.57 - 4.51 (m, 1H), 4.45 - 4.33(m, 2H), 4.28 - 4.16 (m, 2H), 3.03 (s, 3H), 2.51 - 2.38 (m, 1H), 2.35 - 2.20(m, 1H), 1.51 - 1.49 (m, 9H), 1.29 (t, J = 7.2 Hz, 3H), 0.93 (s, 9H), 0.18(d, J = 7.2 Hz, 6H).

[0354] Step 5: Synthesis of ethyl 2-(((tert-butyl)oxy)carbonyl)tetrahydroisoxazole-3-carboxylate

[0355] The starting material 6-(ethoxycarbonyl)-2,2,3,3-tetramethyl-5-(((tert-butyl)oxy)carbonyl)-5-aza-4-oxa-3-silazaoct-8-yl methanesulfonate (1350 mg, 2.96 mmol) was dissolved in tetrahydrofuran (13 mL), cooled to 0°C, and then tetrabutylammonium fluoride (1.3 mL, 4.44 mmol) solution was added. The reaction solution was extracted with ammonium chloride aqueous solution (20 mL) and ethyl acetate (20 mL × 3). The organic phase was washed with sodium bicarbonate aqueous solution (20 mL), saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The crude product obtained by direct concentration of the filtrate was purified by normal-phase silica gel column chromatography to give 605 mg of the product.

[0356] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.75 - 4.61 (m, 1H), 4.24 (d, J =7.2 Hz, 2H), 4.18 - 4.10 (m, 1H), 3.89 - 3.79 (m, 1H), 2.68 - 2.57 (m, 1H),2.52 - 2.41 (m, 1H), 1.50 (s, 9H), 1.30 (s, 3H).

[0357] Step 6: Synthesis of 2-(((tert-butyl)oxy)carbonyl)tetrahydroisoxazole-3-carboxylic acid

[0358] Ethyl 2-(((tert-butyl)oxy)carbonyl)tetrahydroisoxazole-3-carboxylate (605 mg, 2.47 mmol) was dissolved in tetrahydrofuran (5 mL), and a cold aqueous solution of lithium hydroxide monohydrate (404.67 mg, 9.87 mmol) (3 mL) was added. The reaction mixture was stirred at 25 °C for 2 hours. The pH of the reaction mixture was adjusted to approximately 3 by adding 2 mol of potassium hydrogen sulfate aqueous solution, followed by the addition of water (10 mL) and chloroform (30 mL). After separation, the aqueous phase was extracted again with chloroform (20 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated to remove the solvent, yielding 465 mg of the product.

[0359] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.79 - 4.69 (m, 1H), 4.20 - 4.09(m, 1H), 3.91 - 3.83 (m, 1H), 2.70 - 2.60 (m, 2H), 1.53 (s, 9H).

[0360] Step 7: Synthesis of 3-(((4-(4-(methoxycarbonyl)phenyl)phenyl)amino)carbonyl)tetrahydroisoxazole-2-carboxylic acid-tert-butyl ester

[0361] 2-(((tert-butyl)oxy)carbonyl)tetrahydroisoxazole-3-carboxylic acid (400 mg, 1.84 mmol) and methyl 4-(4-aminophenyl)benzoate (460.34 mg, 2.03 mmol) were dissolved in dichloromethane (53 mL), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (458.91 mg, 2.39 mmol) was added. The reaction mixture was stirred at 25 °C for 16 hours. The reaction mixture was then extracted with water (20 mL) and dichloromethane (20 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The crude product obtained by concentration of the filtrate was purified by normal-phase silica gel column chromatography to give 610 mg of the product.

[0362] Step 8: Synthesis of 4-(4-((tetrahydroisoxazole-3-ylcarbonyl)amino)phenyl)benzoate

[0363] 3-(((4-(4-(methoxycarbonyl)phenyl)phenyl)amino)carbonyl)tetrahydroisoxazole-2-carboxylic acid-tert-butyl ester (350 mg, 0.82 mmol) was dissolved in acetonitrile (10 mL), and potassium trimethylsilanolate (421.15 mg, 3.28 mmol) was added. The reaction mixture was stirred at 25 °C for 16 hours. The reaction mixture was extracted with 1 mol hydrochloric acid aqueous solution (20 mL) and ethyl acetate (20 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated under reduced pressure to remove the solvent, yielding 251 mg of the product.

[0364] Step 9: Synthesis of 4-(4-(((2-(((4-(prop-2-yl)phenyl)amino)carbonyl)tetrahydroisoxazol-3-yl)carbonyl)amino)phenyl)benzoic acid

[0365] 4-(4-((tetrahydroisoxazol-3-ylcarbonyl)amino)phenyl)benzoate (251 mg, 0.72 mmol) and triethylamine (0.6 mL, 4.02 mmol) were dissolved in tetrahydrofuran (5 mL), and 4-isocyano-1-(prop-2-yl)benzene (0.10 mL, 0.80 mmol) was added. The reaction solution was reacted at 25 °C for 1 hour. The reaction solution was quenched with methanol (10 mL), the solvent was removed by concentration, and the crude product was separated by high performance liquid chromatography. The fraction was lyophilized to obtain 45 mg of product.

[0366] LC-MS (ESI) m / z: (M+H) + =474.2;

[0367] Step 10: Synthesis of 4-(4-((((3R)-2-(((4-(prop-2-yl)phenyl)amino)carbonyl)tetrahydroisoxazole-3-yl)carbonyl)amino)phenyl)benzoic acid and 4-(4-((((3S)-2-(((4-(prop-2-yl)phenyl)amino)carbonyl)tetrahydroisoxazole-3-yl)carbonyl)amino)phenyl)benzoic acid

[0368] The racemic product 4-(4-(((2-(((4-(prop-2-yl)phenyl)amino)carbonyl)tetrahydroisoxazol-3-yl)carbonyl)amino)phenyl)benzoic acid (45 mg, 0.10 mmol) was purified by supercritical fluid chromatography. Fraction 1 was lyophilized to give compound 13A: 15 mg, retention time: 1.733 min; fraction 2 was lyophilized to give compound 13B: 16 mg, retention time: 2.301 min.

[0369] Compound 13A: LC-MS (ESI): (M+H) + = 474.2;

[0370] 1 H NMR (400 MHz, DMSO-d6) δ = 13.17 - 12.42 (m, 1H), 10.16 (s, 1H), 9.39 (s, 1H), 7.99 (s, 2H), 7.81 (s, 4H), 7.74 (d, J = 8.6 Hz, 2H), 7.55 (d,J = 8.4 Hz, 2H), 7.16 (d, J = 8.4 Hz, 2H), 5.01 - 4.90 (m, 1H), 4.30 - 4.19(m, 1H), 3.91 - 3.76 (m, 1H), 2.88 - 2.78 (m, 1H), 2.72 - 2.63 (m, 1H), 2.47(br d, J = 5.4 Hz, 1H), 1.18 (d, J = 6.8 Hz, 6H).

[0371] Compound 13B: LC-MS (ESI): (M+H) + = 474.2;

[0372] 1H NMR (400 MHz, DMSO-d6) δ = 13.09 - 12.76 (m, 1H), 10.16 (s, 1H), 9.39 (s, 1H), 8.00 (d, J = 8.4 Hz, 2H), 7.89 - 7.77 (m, 4H), 7.77 - 7.70 (m,2H), 7.59 - 7.46 (m, 2H), 7.23 - 7.11 (m, 2H), 5.00 - 4.88 (m, 1H), 4.29 -4.18 (m, 1H), 3.88 - 3.74 (m, 1H), 2.88 - 2.78 (m, 1H), 2.70 - 2.62 (m, 1H), 2.49 - 2.42 (m, 1H), 1.18 (d, J = 6.8 Hz, 6H).

[0373] Example 14: 4-(4-((((2R)-5-oxylidene-1-((2R)-1-oxylidene-1-((4-(prop-2-yl)phenyl)amino)prop-2-yl)pyrrolidine-2-yl)carbonyl)amino)phenyl)benzoic acid, 4-(4-((((2R)-5-oxylidene-1-((2S)-1-oxylidene-1-((4-(prop-2-yl)phenyl)amino)isopropyl)pyrrolidine-2-yl)carbonyl)amino)phenyl)benzoic acid (compounds 14A and 14B))

[0374]

[0375] Step 1: Synthesis of (2R)-5-oxomylidene-2-carboxylic acid benzyl ester

[0376] The starting material (5R)-5-((benzyloxy)carbonyl)-2-oxylidene-1-carboxylic acid tert-butyl ester (3 g, 9.39 mmol) was dissolved in a dioxane solution of hydrochloric acid (50 mL, 2 M). The reaction mixture was stirred at 45 °C for 3 hours. The pH of the reaction mixture was adjusted to 8-9 by adding saturated sodium carbonate aqueous solution, and then diluted with water (100 mL) and ethyl acetate (150 mL). The mixture was separated, and the aqueous phase was extracted three times with ethyl acetate (500 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to remove the solvent, yielding 2.1 g of the product.

[0377] LC-MS (ESI) m / z: (M+H) + =220.0;

[0378] Step 2: Synthesis of tert-butyl 2-((5R)-5-((benzyloxy)carbonyl)-2-oxoylide-1-yl)propionate

[0379] (2R)-5-oxomylidene-2-carboxylic acid benzyl ester (1.8 g, 8.21 mmol) was dissolved in tetrahydrofuran (9 mL), cooled to 0°C, purged with nitrogen, and then sodium hydrogen (0.49 g, 12.32 mmol, 60% purity) was added. After purging with nitrogen three times, the reaction solution was stirred at 0°C for 0.5 hours. Then, a tetrahydrofuran (9 mL) solution of tert-butyl 2-bromopropionate (2.23 g, 10.67 mmol) was added dropwise to the above reaction solution at 0°C. After the addition was complete, the reaction solution was allowed to rise naturally to 20°C and stirred for 3 hours. The reaction solution was quenched with ammonium chloride aqueous solution (100 mL), and ethyl acetate (100 mL) was added for separation. The aqueous phase was extracted three times with ethyl acetate (200 mL). The collected organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated to remove the solvent. The crude product was separated by high performance liquid chromatography, and the fraction was freeze-dried to obtain 790 mg of product.

[0380] Step 3: Synthesis of 2-((5R)-5-((benzyloxy)carbonyl)-2-oxoylidene-1-yl)propionic acid

[0381] 710 mg, 1.47 mmol) of tert-butyl 2-((5R)-5-((benzyloxy)carbonyl)-2-oxoylide-1-yl)propionate was dissolved in dichloromethane (7 mL), and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at 20°C for 1 hour. The solution was diluted with water (10 mL) and dichloromethane (20 mL), and the pH was adjusted to 9 with saturated sodium carbonate solution. The mixture was separated. The aqueous phase was collected, and the pH was adjusted to 3-4 with 1 mol hydrochloric acid solution. The aqueous phase was extracted three times with ethyl acetate (150 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to remove the solvent, yielding 350 mg of the product.

[0382] LC-MS (ESI) m / z: (M+H) + =292.1;

[0383] Step 4: Synthesis of (2R)-5-oxoylide-1-(1-oxoylide-1-((4-(prop-2-yl)phenyl)amino)prop-2-yl)pyrrolidine-2-carboxylic acid benzyl ester

[0384] The starting materials 2-((5R)-5-((benzyloxy)carbonyl)-2-oxomylidene-1-yl)propionic acid (350 mg, 1.20 mmol) and 4-isopropylaniline (195 mg, 1.44 mmol) were dissolved in dichloromethane (7 mL), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (323 mg, 1.68 mmol) was added. The reaction mixture was stirred at 20°C for 12 hours. The reaction mixture was diluted with water (80 mL) and ethyl acetate (100 mL), separated, and the aqueous phase was extracted three times with ethyl acetate (200 mL). The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to remove the solvent. The crude product was purified by normal-phase column chromatography (petroleum ether:ethyl acetate = 1:4, Rf = 0.3), and the fraction was concentrated to remove the solvent to give 145 mg of product.

[0385] LC-MS (ESI) m / z: (M+H) + =409.2;

[0386] Step 5: Synthesis of (2R)-5-oxoylide-1-(1-oxoylide-1-((4-(prop-2-yl)phenyl)amino)prop-2-yl)pyrrolidine-2-carboxylic acid

[0387] The starting material (2R)-5-oxoylide-1-(1-oxoylide-1-((4-(prop-2-yl)phenyl)amino)prop-2-yl)pyrrolidine-2-carboxylic acid benzyl ester (145 mg, 0.36 mmol) was dissolved in methanol (40 mL), and palladium on carbon (147 mg, 0.14 mmol, 10% purity) was added. The reaction mixture was purged with hydrogen three times and stirred at 20°C under a hydrogen (15 Psi) atmosphere for 1 hour. The reaction mixture was directly filtered, and the filter cake was washed with tetrahydrofuran (100 mL), and then washed twice with methanol / dichloromethane (1:1, 200 mL). The filtrate was collected and concentrated to give 140 mg of product.

[0388] LC-MS (ESI) m / z: (M+H) + =319.1;

[0389] Step 6: Synthesis of benzyl benzoate 4-(4-((((2R)-5-oxoylide-1-(1-oxoylide-1-((4-(prop-2-yl)phenyl)amino)prop-2-yl)pyrrolidine-2-yl)carbonyl)amino)phenyl)benzoate

[0390] The starting materials (2R)-5-oxoylide-1-(1-oxoylide-1-((4-(propyl-2-yl)phenyl)amino)propyl-2-yl)pyrrolidine-2-carboxylic acid (140 mg, 0.44 mmol) and benzyl 4-(4-aminophenyl)benzoate (160 mg, 0.53 mmol) were dissolved in dichloromethane (8 mL), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (118 mg, 0.62 mmol) was added. The reaction mixture was stirred at 20°C for 2 hours. The mixture was diluted with water (50 mL) and ethyl acetate (50 mL), separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL). The organic phase was collected, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and concentrated to remove the solvent, yielding 300 mg of crude product.

[0391] LC-MS (ESI) m / z: (M+H) + =604.4;

[0392] Step 7: Synthesis of 4-(4-((((2R)-5-oxoylide-1-(1-oxoylide-1-((4-(prop-2-yl)phenyl)amino)prop-2-yl)pyrrolidine-2-yl)carbonyl)amino)phenyl)benzoic acid

[0393] Benzyl benzoate (300 mg, 0.30 mmol) was dissolved in tetrahydrofuran (30 mL), and palladium on carbon (300 mg, 2.82 mmol, 10% purity) was added. The reaction mixture was purged with hydrogen three times and stirred at 20°C for 1 hour under a 15 Psi hydrogen atmosphere. The reaction mixture was directly filtered, and the filter cake was washed with tetrahydrofuran (150 mL), followed by washing three times with methanol / dichloromethane (1:1, 300 mL). The filtrate was collected and concentrated to obtain the crude product. The crude product was separated by high performance liquid chromatography, and the fraction was lyophilized to obtain 110 mg of the product.

[0394] LC-MS (ESI) m / z: (M+H) + =514.3;

[0395] Step 8: Synthesis of 4-(4-((((2R)-5-oxylidene-1-((2R)-1-oxylidene-1-((4-(prop-2-yl)phenyl)amino)prop-2-yl)pyrrolidine-2-yl)carbonyl)amino)phenyl)benzoic acid and 4-(4-((((2R)-5-oxylidene-1-((2S)-1-oxylidene-1-((4-(prop-2-yl)phenyl)amino)prop-2-yl)pyrrolidine-2-yl)carbonyl)amino)phenyl)benzoic acid)

[0396] The raw material 4-(4-((((2R)-5-oxoylide-1-(1-oxoylide-1-((4-(prop-2-yl)phenyl)amino)prop-2-yl)pyrrolidine-2-yl)carbonyl)amino)phenyl)benzoic acid (130 mg, 0.25 mmol) was purified by supercritical fluid chromatography. Fraction 1 was lyophilized to obtain product 14A: 45 mg, retention time: 1.887 min; fraction 2 was lyophilized to obtain product 14B: 55 mg, retention time: 2.061 min.

[0397] Compound 14A: LC-MS (ESI) m / z: (M+H) + =514.3;

[0398] 1 H NMR (400 MHz, DMSO-d6) δ = 12.93 (s, 1H), 10.53 (s, 1H), 10.13 (s,1H), 8.00 (d, J = 8.4 Hz, 2H), 7.78 - 7.73 (m, 6H), 7.51 (d, J = 8.4 Hz, 2H),7.18 (d, J = 8.8 Hz, 2H), 4.72 (d, J = 8.4 Hz, 1H), 4.62 (q, J = 7.2 Hz, 1H),2.86 - 2.82 (m, 1H), 2.46 - 2.39 (m, 2H), 2.27 - 2.21 (m, 1H), 2.07 - 2.02(m, 1H), 1.31 (d, J = 7.6 Hz, 3H), 1.18 (d, J = 6.8 Hz, 6H).

[0399] Compound 14B: LC-MS (ESI) m / z: (M+H) + =514.3;

[0400] 1H NMR (400 MHz, DMSO-d6) δ = 12.92 (s, 1H), 10.78 (s, 1H), 10.20 (s,1H), 7.99 (d, J = 7.6 Hz, 2H), 7.78 - 7.73 (m, 6H), 7.47 (d, J = 8.0 Hz, 2H),7.14 (d, J = 8.0 Hz, 2H), 4.58 - 4.51 (m, 2H), 2.83 - 2.80 (m, 1H), 2.43 -2.41 (m, 2H), 2.29 - 2.25 (m, 1H), 2.08 - 2.04 (m, 1H), 1.38 (d, J = 6.8 Hz,3H), 1.15 (d, J = 6.4 Hz, 6H).

[0401] Biological test data

[0402] Experimental Example 1: Determination of the inhibitory activity of the compounds of the present invention against glucose-dependent insulinotropic peptide receptors.

[0403] Experimental objective: To determine the antagonistic activity of the tested compounds against hGIPR using a stable cell line of CHO-K1 (a subclone of Chinese hamster ovary cells) expressing human GIPR (hGIPR).

[0404] Experimental Protocol: This assay evaluates the in vitro antagonistic activity of test compounds by detecting the level of cyclic adenosine monophosphate (cAMP) in CHO-K1 cells stably expressing hGIPR. Upon agonist activation, hGIPR binds to the G protein complex, triggering a series of signal transductions to regulate changes in intracellular cAMP levels. Therefore, measuring intracellular cAMP levels can be used for pharmaceutical characterization. Intracellular cAMP levels were quantified using HTRF (homogeneous time-resolved fluorescence) technology. The principle of this method is that intracellularly produced cAMP competitively binds to cAMP labeled with the receptor dye Eu. TM The labeled cAMP antibody produces a specific signal that is inversely proportional to the intracellular cAMP level.

[0405] The test compound was dissolved in 100% DMSO (dimethyl sulfoxide) and serially diluted 4-fold, for a total of 10 dilutions. Each serially diluted test compound (50 nL) was transferred to a 384-well plate in duplicate. The final test concentration range of the compound was 38 pM to 10 μM, with a final DMSO concentration of 0.5%.

[0406] cAMP Hunter TMAfter resuscitation, passage, and expansion of CHO-K1 GIPR Gs Cell Lines, the culture supernatant was removed, and the cells were washed with DPBS. Following trypsin treatment and cell counting, the cells were resuspended in Stimulation buffer (1×HBSS, 20 mM HEPES, 0.1% BSA, 200 μM IBMX). 5 μL of cell suspension (2000 cells / well) was added to a 384-well plate containing the test compound, mixed, centrifuged at 600 rpm for 3 min, and incubated at 37°C for 2 h. Subsequently, 5 μL of GIPR agonist (0.5 nM hGIP) was added. 1-42 Incubate at 37°C for 30 min. Finally, add 5 μL of Eu-cAMP solution and 5 μL of ULight according to the cAMP Kit instructions. TM -anti-cAMP solution, incubated at room temperature in the dark for 1 h. Fluorescence signals of the samples were acquired using an EnVision multi-functional microplate reader (excitation wavelength: 320 nM; emission wavelength: 665 nM / 615 nM).

[0407] Data were analyzed by the ratio of fluorescence intensity at 665 nM and 615 nM for each well sample, and extrapolated from the signal values ​​of a cAMP standard curve at known concentrations. The data for each well cell were expressed in nanomolar (nM) cAMP. The % effect of the test compound was calculated using the formula: % = 100 - (cAMP level of the test compound - cAMP level at 100% effect) / (cAMP level at 0% effect - cAMP level at 100% effect) × 100. A 0% effect was defined as 0.5 nM hGIP. 1-42 The intracellular cAMP level produced after treatment, with a 100% effect defined as 0.5 nM hGIP. 1-42 and 20 μM hGIPR antagonist hGIP 3-42 Intracellular cAMP levels after mixed treatment. The percentage effect at each concentration of the test compound was plotted using a four-parameter logistic dose-response equation, and the IC50 was calculated. 50 value.

[0408] The results are shown in Table 1.

[0409] Table 1. Antagonistic activity of compounds against hGIPR

[0410]

[0411] Conclusion: The compounds of this invention have significant antagonistic activity against GIP receptors.

[0412] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

Claims

1. A compound, which is a compound of formula (I), or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of a compound of formula (I); (I)； in: L1 is , , , , phenyl or 5-6 heteroaryl; L2 is a bond or -(CR7R8) n -; Each R1 is independently selected from H, D, F, Cl, Br, I, NH2, CN, OH, guanidinyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups; R2 is -OH, - , -B(OH)2, -C(=O)OW1, -C(=O)NW2W3 or -S(=O)2W4, where W1, W2, W3 and W4 are independently selected from H, D, C respectively. 1-6 Alkyl, C 3-8 cycloalkyl and C 6-10 Aryl, the C 1-6 Alkyl, C 3-8 cycloalkyl and C 6-10 The aryl group can be independently and optionally replaced by one, two, or three substituents selected from D, F, Cl, Br, I, NH2, CN, OH, and guanidinyl groups; Ring A is , Or C 5-10 Bridged heterocyclic alkyl groups; Ring B is C 6-10 Aryl or 5-12 heteroaryl, wherein C 6-10 The aryl and 5-12 heteroaryl groups can be independently and optionally surrounded by 1, 2, 3 or 4 groups selected from D, F, Cl, Br, I, oxo (=O), NH2, CN, OH, guanidinyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 1-6 Substituents of cycloalkyl and 3-6 membered heterocyclic alkyl groups; Ring C and ring D are independently selected from phenyl, naphthyl, 5-6 membered heteroaryl, and C64, respectively. 3-6 Monocyclic cycloalkyl, 3-6 membered monocyclic heterocyclic alkyl, C 5-10 Spirocycloalkyl, C 5-10 cycloalkyl, C 5-10 Bridged cycloalkyl, 5-10 membered spirocyclic heterocyclic alkyl, 5-10 membered fused heterocyclic alkyl and 5-10 membered bridged cycloalkyl, wherein the phenyl, naphthyl, 5-6 membered heteroaryl, C 3-6 Monocyclic cycloalkyl, 3-6 membered monocyclic heterocyclic alkyl, C 5-10 Spirocycloalkyl, C 5-10 cycloalkyl, C 5-10 Bridged cycloalkyl, 5-10 membered spirocyclic heterocyclic alkyl, 5-10 membered fused heterocyclic alkyl, and 5-10 membered bridged cycloalkyl can be independently and optionally surrounded by 1, 2, 3, or 4 groups selected from D, F, Cl, Br, I, oxo (=O), NH2, CN, OH, guanidinyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 1-6 Substituents of cycloalkyl and 3-6 membered heterocyclic alkyl groups; R3, R4, R5, and R6 are independently selected from H, D, F, Cl, Br, I, CN, guanidino, and C, respectively. 1-6 Alkyl and C 1-6 Alkoxy; the C 1-6 Alkyl and C 1-6 The alkoxy group can be optionally and independently substituted by 1, 2, 3, 4 or 5 substituents selected from D, F, Cl, Br, I; Each of R7 and R8 is independently selected from H, D, F, Cl, Br, I, OH, and C. 1-6 Alkyl and C 1-6 Alkoxy, the C 1-6 Alkyl and C 1-6 The alkoxy group may be independently and optionally substituted by 1, 2, 3, 4 or 5 substituents selected from H, D, F, Cl, Br, I, oxo (=O), CN, NH2 and OH; or R7 and R8 may optionally form a C with the carbon atom they are attached to. 3-6 cycloalkyl, the C 3-6 The cycloalkyl group may be independently and optionally substituted by one, two or three substituents selected from D, oxo (=O), F, Cl, Br, I, CN, NH2 and OH; m can be 0, 1, 2, 3, or 4; n is 1, 2, or 3.

2. The compound according to claim 1, characterized in that: Each R1 is independently selected from H, D, F, Cl, Br, I, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3, and -OCH2CF2CHF2.

3. The compound according to claim 1, characterized in that: Ring A is , , , , , , , or , , or .

4. The compound according to claim 1, characterized in that: L1 is , , , , phenyl, , , , , , , , , , or .

5. The compound according to claim 1, characterized in that: Ring B is selected from any of the following groups: phenyl, , , , , , , , , , , , , , , , , , , , , , , , , , , , and ; The above groups may be independently and optionally replaced by 1, 2 or 3 substituents selected from D, oxo (=O), F, Cl, Br, I, CN, NH2, OH, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3 and -OCH2CF2CHF2.

6. The compound according to claim 1, characterized in that: Rings C and D are independently selected from phenyl, naphthyl, and phenyl groups, respectively. , , , , , , , , , , , , , , , , , , , , , , , , , and ; The above groups may be independently and optionally replaced by 1, 2 or 3 substituents selected from D, oxo (=O), F, Cl, Br, I, CN, NH2, OH, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3 and -OCH2CF2CHF2.

7. The compound according to claim 1, characterized in that: R3, R4, R5, and R6 are independently selected from H, D, F, Cl, Br, I, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3, and -OCH2CF2CHF2; R7 and R8 are independently selected from H, D, F, Cl, Br, I, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, -CHF2, -CF3, -CHFCH2F, -CF2CHF2, -CH2CF3, -CH2CF2CHF2, methoxy, ethoxy, n-propoxy, isopropoxy, -OCHF2, -OCF3, -OCHFCH2F, -OCF2CHF2, -OCH2CF3, and -OCH2CF2CHF2; or R7 and R8 may optionally form cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl with the carbon atom they are connected to, wherein the cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl may be independently and optionally substituted by 1, 2, or 3 substituents selected from D, F, Cl, Br, and I.

8. A compound having one of the following structures, or a stereoisomer, geometric isomer, tautomer, nitride, hydrate, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 9. A pharmaceutical composition comprising the compound of any one of claims 1-8; the pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient, carrier, adjuvant, or any combination thereof.

10. Use of the compound of any one of claims 1-8 or the pharmaceutical composition of claim 9 in the preparation of a medicament for the prevention, treatment or relief of a patient’s GIP receptor antagonist-mediated disease.

11. The use according to claim 10, wherein, The diseases mediated by the GIP receptor antagonists are obesity, type 2 diabetes, pulmonary deposition disease, non-alcoholic steatohepatitis, atherosclerosis, heart failure, polycystic ovary syndrome, Alzheimer's disease, or osteoporosis.

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