Novel benzazepine spirocyclic derivatives

Abstract

The benzazaspiro derivatives represented by formula (I) and pharmaceutically acceptable salts thereof, as well as the compounds, can be used for diagnosing, preventing and / or treating diseases associated with vasopressin receptors.

Description

[0001] The present invention claims the following priority:

[0002] Application number: CN202011353057.1, filing date: November 26, 2020;

[0003] Application number: CN202111322711.7, application date: November 9, 2021. Technical Field

[0004] The present invention relates to novel benzazepine spirocyclic derivatives and salts thereof. The present invention also relates to drugs containing the benzazepine spirocyclic derivatives and salts thereof as active ingredients, which can be used for diagnosing, preventing and / or treating diseases related to vasopressin receptors. Background Art

[0005] Hormones play a crucial role in regulating human internal homeostasis. Arginine vasopressin (AVP) is closely linked to the regulation of water and sodium metabolism. Disturbances in AVP metabolism can cause a variety of conditions, including hyponatremia, syndrome of inappropriate antidiuretic hormone secretion, congestive heart failure, cirrhosis, kidney disease, hypertension, and edema. Arginine vasopressin (AVP) receptor antagonists inhibit the binding of AVP to the receptor, thereby providing therapeutic benefits for these conditions. Arginine vasopressin V2 receptor antagonists, such as tolvaptan, can increase free water excretion without affecting electrolyte metabolism, making them ideal agents for treating these conditions. However, marketed AVP V2 receptor antagonists, such as tolvaptan, are metabolized by hepatic enzymes, producing a large number of metabolites in the body and leading to severe drug-induced hepatotoxicity. The FDA has issued a black box warning on the drug's label, limiting its application. Therefore, the development of novel V2 receptor antagonists with high efficacy and minimal side effects is crucial. Summary of the Invention

[0006] One object of the present invention is to provide a novel benzazepine spirocyclic compound or a salt thereof having vasopressin V2 receptor antagonism, favorable metabolic stability and / or metabolites with reduced drug-induced hepatotoxicity, and the medical use of the compound.

[0007] In one aspect of the present invention, the present invention provides a compound represented by formula (X), its optical isomers and pharmaceutically acceptable salts thereof,

[0008]

[0009] in,

[0010] Ring A is selected from heterocycloalkyl and cycloalkyl, wherein the heterocycloalkyl and cycloalkyl are optionally substituted by 1, 2, 3 or 4 R A replace;

[0011] Ring B is selected from aryl, heteroaryl, heterocycloalkyl and cycloalkyl, wherein the aryl, heteroaryl, heterocycloalkyl or cycloalkyl is optionally substituted with 1, 2 or 3 R3;

[0012] Ring C is selected from aryl, heteroaryl, heterocycloalkyl and cycloalkyl, said aryl, heteroaryl, heterocycloalkyl or cycloalkyl being optionally substituted with 1, 2 or 3 R4;

[0013] T1 and T2 are independently selected from N and CH;

[0014] R1, R2, R3, R4 are each independently selected from H, F, Cl, Br, I, CN, OH, NH2, alkyl, heteroalkyl, aryl, heteroaryl, heterocycloalkyl and cycloalkyl, wherein the alkyl, heteroalkyl, aryl, heteroaryl, heterocycloalkyl or cycloalkyl is optionally substituted with 1, 2, 3 or 4 R;

[0015] R and R A R' is independently selected from H, F, Cl, Br, I, CN, OH, NH2, alkyl, heteroalkyl, aryl, heteroaryl, heterocycloalkyl and cycloalkyl, wherein the alkyl, heteroalkyl, aryl, heteroaryl, heterocycloalkyl or cycloalkyl is optionally substituted with 1, 2, 3 or 4 R's;

[0016] R' is selected from H, F, Cl, Br, I, CN, OH, NH2, alkyl and heteroalkyl;

[0017] m1 and m2 are independently selected from 1, 2, 3 or 4;

[0018] L X is selected from -NH(C=O)-, -alkyl-NH(C=O)-, -NH(C=O)-alkyl-, alkyl, alkenyl, and alkynyl, said -alkyl-NH(C=O)-, -NH(C=O)-alkyl-, alkyl, alkenyl, or alkynyl being optionally substituted with 1, 2, 3, or 4 R;

[0019] Furthermore, when ring A is selected from heterocycloalkyl, the compound represented by formula (I) is not selected from

[0020] The heterocycloalkyl or heteroaryl group contains 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -O-, -NH-, -N=, -S-, -C(=O)-, -C(=O)O-, -S(=O)-, -S(=O)2- and N.

[0021] In another aspect of the present invention, the present invention provides a compound represented by formula (I), an optical isomer thereof and a pharmaceutically acceptable salt thereof,

[0022]

[0023] in,

[0024] Ring A is selected from 3-6 membered heterocycloalkyl and C 3-6 Cycloalkyl, the 3-6 membered heterocycloalkyl and C 3-6 The cycloalkyl group is optionally substituted with 1 or 2 R A replace;

[0025] Ring B is selected from phenyl and 5-6 membered heteroaryl, wherein the phenyl or 5-6 membered heteroaryl is optionally substituted with 1, 2 or 3 R3;

[0026] Ring C is selected from phenyl and 5-6 membered heteroaryl, wherein the phenyl or 5-6 membered heteroaryl is optionally substituted with 1, 2 or 3 R4;

[0027] T1 and T2 are independently selected from N and CH;

[0028] R1 is independently selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups;

[0029] R2 is independently selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl and C 3-6 Cycloalkyl, the C 1-6 Alkyl or C 3-6 Cycloalkyl is optionally substituted with 1, 2 or 3 R;

[0030] R3 are independently selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups;

[0031] R4 are independently selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 3-6 Cycloalkyl, phenyl and 5-6 membered heteroaryl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 3-6 Cycloalkyl, phenyl or 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 R;

[0032] R and R A are independently selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Alkylamino, the C 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Alkylamino is optionally substituted with 1, 2 or 3 R';

[0033] R' is selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 alkyl;

[0034] m1 and m2 are independently selected from 1, 2 or 3;

[0035] Furthermore, when ring A is selected from 3-6 membered heterocycloalkyl, the compound represented by formula (I) is not selected from

[0036] The 3-6 membered heterocycloalkyl or 5-6 membered heteroaryl contains 1, 2 or 3 heteroatoms or heteroatom groups independently selected from -O-, -NH-, -N=, -S-, -C(=O)-, -C(=O)O-, -S(=O)-, -S(=O)2- and N.

[0037] In another aspect of the present invention, the present invention provides a compound represented by formula (II), an optical isomer thereof and a pharmaceutically acceptable salt thereof,

[0038]

[0039] in,

[0040] X1 is selected from C(R A )2, NH and O;

[0041] X2 is selected from CH and N;

[0042] T1 and T2 are independently selected from N and CH;

[0043] R1 is independently selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups;

[0044] R 2a and R 2b are independently selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl and C 3-6 Cycloalkyl, the C 1-6Alkyl or C 3-6 Cycloalkyl is optionally substituted with 1, 2 or 3 R;

[0045] R3 is selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups;

[0046] R4 are independently selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 3-6 Cycloalkyl, phenyl and 5-6 membered heteroaryl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 3-6 Cycloalkyl, phenyl or 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 R;

[0047] R is selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Alkylamino, the C 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Alkylamino is optionally substituted with 1, 2 or 3 R';

[0048] R' is selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 alkyl;

[0049] n1 is selected from 0, 1 or 2;

[0050] n2 is selected from 1, 2 or 3;

[0051] Furthermore, when X1 is selected from O, the compound represented by formula (II) is not selected from

[0052] In another aspect of the present invention, the present invention also provides a compound represented by formula (III), an optical isomer thereof and a pharmaceutically acceptable salt thereof,

[0053]

[0054] in,

[0055] R1 is independently selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups;

[0056] R3 is selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups;

[0057] R4 are independently selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 3-6 Cycloalkyl, phenyl and 5-6 membered heteroaryl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 3-6 Cycloalkyl, phenyl or 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 R;

[0058] R is selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Alkylamino, the C 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Alkylamino is optionally substituted with 1, 2 or 3 R';

[0059] R' is selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 alkyl;

[0060] X2 is selected from CH and N.

[0061] In some embodiments of the present invention, R is selected from H, F, Cl, Br, I, CN, OH, NH2, CH3, CF3, Other variables are as defined in the present invention.

[0062] In some embodiments of the present invention, R A is selected from H, OH and NH2, and other variables are as defined in the present invention.

[0063] In some embodiments of the present invention, R4 is selected from H, F, Cl, Br, I, CN, OH, NH2, CH3, CF3, cyclopropyl, cyclobutyl, cyclopentyl, phenyl, pyridyl, pyrimidinyl, thienyl and thiazolyl, and other variables are as defined herein.

[0064] In some embodiments of the present invention, ring A is selected from cyclopropyl, cyclobutyl, cyclopentyl, aziridine, oxiranyl, azetidinyl, oxetane, pyrrolidinyl and tetrahydrofuranyl, and the cyclopropyl, cyclobutyl, cyclopentyl, aziridine, oxiranyl, azetidinyl, oxetane, pyrrolidinyl or tetrahydrofuranyl is optionally replaced by 1 or 2 R A Substitution, other variables are as defined in the present invention.

[0065] In some embodiments of the present invention, ring A is selected from Other variables are as defined in the present invention.

[0066] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0067] In some embodiments of the present invention, Ring B is selected from phenyl and pyridinyl, wherein the phenyl or pyridinyl is optionally substituted with 1, 2 or 3 R3, and other variables are as defined herein.

[0068] In some embodiments of the present invention, ring C is selected from Other variables are as defined in the present invention.

[0069] In another aspect of the present invention, the present invention also provides the following compounds, their optical isomers and pharmaceutically acceptable salts, which are selected from

[0070]

[0071] In another aspect of the present invention, the present invention also provides the use of the aforementioned compound, its optical isomers and pharmaceutically acceptable salts thereof in the preparation of a medicament for preventing or treating diseases related to the arginine vasopressin V1a receptor, arginine vasopressin V1b receptor, arginine vasopressin V2 receptor, sympathetic nervous system or renin-angiotensin-aldosterone system.

[0072] In some embodiments of the present invention, the diseases associated with arginine vasopressin V1a receptor, arginine vasopressin V1b receptor, arginine vasopressin V2 receptor, sympathetic nervous system or renin-angiotensin-aldosterone system include: hypertension, Reye's syndrome, dysmenorrhea, premature birth, corticotropin-releasing hormone secretion disorder, adrenal hyperplasia, depression, chronic congestive heart failure, cirrhosis, syndrome of inappropriate antidiuretic hormone secretion, hyponatremia caused by chronic heart failure / cirrhosis / inappropriate antidiuretic hormone secretion, or polycystic kidney disease.

[0073] Definition and Description

[0074] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0075] As used herein, the phrase "at least one" when referring to a list of one or more elements should be understood to mean at least one element selected from any one or more elements in the list of elements, but does not necessarily include at least one of each element specifically listed in the list of elements, and does not exclude any combination of elements in the list of elements. This definition also allows that elements other than the elements specifically identified in the list of elements to which the phrase "at least one" refers may optionally be present, whether related or unrelated to those specifically identified elements.

[0076] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0077] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, trifluoroacetic acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine) and organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and can be converted into either base or acid addition salts.

[0078] The pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of the two.

[0079] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. All such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, are contemplated by the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0080] The compounds of the present invention may exist in specific forms. Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers with different functional groups that are in dynamic equilibrium at room temperature and can quickly convert into each other. If tautomerism is possible (such as in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also known as prototropic tautomers) include interconversions that occur through proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence isomers (valencetautomers) include interconversions that occur through the reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-ene-2-one.

[0081] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14C). For example, deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared to undeuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced therapeutic efficacy, and prolonged biological half-life. All isotopic variations of the compounds of this invention, whether radioactive or not, are encompassed by this invention. "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs as well as instances where it does not.

[0082] When the group valence bond is marked with a dotted line When, for example, In the example, the dashed line represents the point of attachment of the group to the rest of the molecule. When, for example, In the example, the dotted line represents a single bond or its absence, which also means Represents a single bond or double bond

[0083] The term "substituted" or "substituted with" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, including deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. The term "optionally substituted" or "optionally substituted with" means that the atom may or may not be substituted. Unless otherwise specified, the type and number of substituents may be any based on chemical practicability.

[0084] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 1, 2, or 3 R's, the group may optionally be substituted with 1, 2, or 3 R's, with each occurrence of R' being an independent choice. Furthermore, combinations of substituents and / or their variants are permitted only if such combinations result in stable compounds.

[0085] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected, such as When L1 represents a single bond, it means that the structure is actually

[0086] When the substituents listed do not specify through which atom they are connected to the substituted group, such substituents can be bonded through any atom thereof. For example, a pyridyl substituent can be connected to the substituted group through any carbon atom on the pyridine ring.

[0087] When the linking group is listed without specifying its linking direction, its linking direction is arbitrary, for example, The connecting group L is -CH2O-, in which case -CH2O- can connect phenyl and cyclopentyl in the same direction as reading from left to right to form It is also possible to connect phenyl and cyclopentyl groups in the opposite direction of reading from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0088] Unless otherwise specified, the number of atoms in a ring is generally defined as the number of members of the ring, for example, a "3-6 membered ring" refers to a "ring" having 3-6 atoms arranged around it.

[0089] Unless otherwise specified, the term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and even more preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched-chain isomers thereof. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo.

[0090] Unless otherwise specified, the term “C 1-6"Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. 1-6 Alkyl groups include C 1-5 、C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 , C6 and C5 alkyl, etc.; which can be monovalent (such as CH3), divalent (-CH2-) or polyvalent (such as ). C 1-6 Examples of alkyl groups include, but are not limited to, CH3, wait.

[0091] Unless otherwise specified, the term “C 1-4 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 4 carbon atoms. 1-4 Alkyl groups include C 1-2 、C 1-3 、C 3-4 and C 2-3 Alkyl, etc.; it can be monovalent (such as CH3), divalent (-CH2-) or polyvalent (such as ). C 1-4 Examples of alkyl groups include, but are not limited to, CH3, wait.

[0092] Unless otherwise specified, “C 2-3 "Alkenyl" is used to refer to a linear or branched hydrocarbon group consisting of 2 to 3 carbon atoms containing at least one carbon-carbon double bond, which may be located at any position of the group. 2-3 Alkenyl includes C3 and C2 alkenyl; the C 2-3 Alkenyl groups can be monovalent, divalent, or polyvalent. 2-3 Examples of alkenyl groups include, but are not limited to wait.

[0093] Unless otherwise specified, “C 2-3 "Alkynyl" is used to represent a linear or branched hydrocarbon group consisting of 2 to 3 carbon atoms containing at least one carbon-carbon triple bond, which may be located at any position of the group. It may be monovalent, divalent or polyvalent. The C 2-3 Alkynyl groups include C3 and C2 alkynyl groups. 2-3 Examples of alkynyl groups include, but are not limited to wait.

[0094] Unless otherwise specified, the term "heteroalkyl" by itself or in combination with another term refers to a stable straight or branched chain alkyl radical or combination thereof consisting of a certain number of carbon atoms and at least one heteroatom or heteroatom group. In some embodiments, the heteroatom is selected from B, O, N and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen heteroatom is optionally quaternized. In other embodiments, the heteroatom group is selected from -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O)2-, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O)2N(H)- and -S(=O)N(H)-. In some embodiments, the heteroalkyl group is C 1-6 In other embodiments, the heteroalkyl group is C 1-3 Heteroalkyl. The heteroatom or heteroatom group may be located at any interior position of the heteroalkyl group, including the position at which the alkyl group is attached to the remainder of the molecule, but the term "alkoxy" is used conventionally to refer to those alkyl groups that are attached to the remainder of the molecule through an oxygen atom. Examples of heteroalkyl groups include, but are not limited to, -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH2(CH3)2, -CH2-CH2-O-CH3, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)(CH2CH3), -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH2(CH3)2, -CH2-S-CH2-CH3, -CH2-CH2, -S(=O)-CH3, -CH2-CH2-S(=O)2-CH3, and up to two heteroatoms can be consecutive, for example, -CH2-NH-OCH3.

[0095] Unless otherwise specified, the term “C 1-6 "Alkoxy" refers to an alkyl group containing 1 to 6 carbon atoms which is attached to the rest of the molecule via an oxygen atom. 1-6 Alkoxy groups include C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 , C6, C5, C4 and C3 alkoxy, etc. 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), pentoxy (including n-pentoxy, isopentoxy and neopentoxy), hexyloxy, and the like.

[0096] Unless otherwise specified, the term “C1-3 "Alkoxy" refers to those alkyl groups containing 1 to 3 carbon atoms which are attached to the rest of the molecule via an oxygen atom. The C 1-3 Alkoxy groups include C 1-3 、C 1-2 、C 2-3 , C1, C2 and C3 alkoxy, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

[0097] Unless otherwise specified, the term “C 1-6 "Alkylamino" means an alkyl group containing 1 to 6 carbon atoms which is attached to the rest of the molecule via an amino group. 1-6 Alkylamino groups include C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 , C6, C5, C4, C3 and C2 alkylamino, etc. 1-6 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -N(CH2CH3)(CH2CH3), -NHCH2CH2CH3, -NHCH2(CH3)2, -NHCH2CH2CH2CH3, and the like.

[0098] Unless otherwise specified, the term “C 1-3 "Alkylamino" means an alkyl group containing 1 to 3 carbon atoms which is attached to the rest of the molecule via an amino group. 1-3 Alkylamino groups include C 1-3 、C 1-2 、C 2-3 , C1, C2 and C3 alkylamino, etc. 1-3 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -NHCH2CH2CH3, -NHCH2(CH3)2, and the like.

[0099] Unless otherwise specified, the term “C 1-6 "Alkylthio" refers to those alkyl groups containing 1 to 6 carbon atoms which are linked to the rest of the molecule via a sulfur atom. 1-6 Alkylthio includes C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 , C6, C5, C4, C3 and C2 alkylthio, etc. 1-6Examples of alkylthio groups include, but are not limited to, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH2(CH3)2, and the like.

[0100] Unless otherwise specified, the term “C 1-3 "Alkylthio" refers to those alkyl groups containing 1 to 3 carbon atoms which are linked to the rest of the molecule via a sulfur atom. 1-3 Alkylthio includes C 1-3 、C 1-2 、C 2-3 , C1, C2 and C3 alkylthio, etc. 1-3 Examples of alkylthio groups include, but are not limited to, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH2(CH3)2, and the like.

[0101] Unless otherwise specified, the term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms (which can be a specific point or an interval consisting of two points, such as 3, 4, 5, 6 ring atoms, 4 to 11 ring atoms, 6 to 12 ring atoms, etc.), more preferably 3 to 8 carbon atoms, and most preferably 3 to 6 (e.g., 3, 4, 5 or 6) carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc., preferably cycloalkyl; polycyclic cycloalkyls include spirocyclic, fused ring and bridged ring cycloalkyls.

[0102] The term "spiroalkyl" refers to a polycyclic group having a carbon atom (called a spiro atom) shared between 5 to 20 monocyclic rings, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiro atoms shared between the rings, the spiroalkyl group is divided into a single spiroalkyl group, a double spiroalkyl group or a multi-spiroalkyl group, preferably a single spiroalkyl group and a double spiroalkyl group. More preferably, it is a 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiroalkyl group. Non-limiting examples of spiroalkyl groups include: wait.

[0103] The term "fused cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with the other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl groups. Non-limiting examples of fused cycloalkyl groups include: wait.

[0104] The term "bridged cycloalkyl" refers to a 5-20 membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 5-14 members, more preferably 7-10 members. Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl group, preferably a bicyclic, tricyclic or tetracyclic group, more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged cycloalkyl groups include: wait.

[0105] The cycloalkyl group includes the above-mentioned cycloalkyl groups (such as monocyclic, condensed, spirocyclic and bridged cycloalkyl groups) fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl and the like; preferably phenylcyclopentyl and tetrahydronaphthyl.

[0106] The cycloalkyl group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo.

[0107] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, but excluding the ring portion of -OO-, -OS- or -SS-, the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms (which can be specific points or an interval consisting of any two points, such as 3, 4, 5, 6 ring atoms, 4 to 11 ring atoms, 6 to 12 ring atoms, etc.), of which 1 to 4 are heteroatoms; preferably, it contains 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; more preferably, it contains 3 to 6 ring atoms, of which 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include azetidinyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc., preferably tetrahydropyranyl, piperidinyl, pyrrolidinyl, etc. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups.

[0108] The term "spiroheterocyclyl" refers to a polycyclic heterocyclic group in which the monocyclic rings of 5 to 20 members share one atom (called a spiro atom), wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer 0 to 2) heteroatom, and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated π electron system. It is preferably 6 to 14 members, more preferably 7 to 11 members. According to the number of shared spiral atoms between the rings, the spiro heterocyclic group is divided into a single spiral heterocyclic group, a double spiral heterocyclic group or a multi-spiro heterocyclic group, preferably a single spiral heterocyclic group and a double spiral heterocyclic group. More preferably 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiral heterocyclic group. Non-limiting examples of spiro heterocyclic groups include: wait.

[0109] The term "fused heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 20 members, wherein each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, the remaining ring atoms being carbon. Preferably, it is 6 to 14 members, more preferably 7 to 11 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of fused heterocyclic groups include: wait.

[0110] The term "bridged heterocyclyl" refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected, which may contain one or more double bonds but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, the remaining ring atoms being carbon. Preferably, it is 6 to 14 members, more preferably 7 to 11 members. Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic group, preferably a bicyclic, tricyclic or tetracyclic group, more preferably a bicyclic or tricyclic group. Non-limiting examples of bridged heterocyclic groups include: wait.

[0111] The heterocyclic group includes the above-mentioned heterocyclic groups (such as monocyclic, fused, spirocyclic and bridged heterocyclic groups) fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, and its non-limiting examples include: wait.

[0112] The heterocyclic group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo.

[0113] The term "aryl" refers to a 6- to 20-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, more preferably 6-membered, such as phenyl and naphthyl. The aryl group includes the above-mentioned aryl groups fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, non-limiting examples of which include:

[0114] The aryl group may be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from one or more substituents of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio.

[0115] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 20 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 5-10-membered and contains 1 to 3 heteroatoms; more preferably 5-membered or 6-membered and contains 1 to 3 heteroatoms; non-limiting examples include pyrazolyl, imidazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, thiadiazole, pyrazinyl and the like. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0116] The heteroaryl group may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from one or more substituents of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio and heterocycloalkylthio.

[0117] Unless otherwise specified, “C 3-6 "Cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which is a monocyclic or bicyclic ring system. 3-6 Cycloalkyl groups include C 3-5、C 4-5 and C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0118] Unless otherwise specified, the term "3-6 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 3 to 6 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p , p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spirocyclic, fused and bridged rings. In addition, with respect to the "3-6 membered heterocycloalkyl", a heteroatom may occupy the position at which the heterocycloalkyl is connected to the rest of the molecule. The 3-6 membered heterocycloalkyl includes 4-6 membered, 5-6 membered, 4 membered, 5 membered and 6 membered heterocycloalkyls, etc. Examples of 3-6 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl or homopiperidinyl, etc.

[0119] Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" are used interchangeably in the present invention. The term "5-6 membered heteroaryl" refers to a monocyclic group with a conjugated π electron system consisting of 5 to 6 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) p, p is 1 or 2). 5-6 membered heteroaryl can be connected to the rest of the molecule through a heteroatom or a carbon atom. The 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl. Examples of the 5-6 membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl) and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

[0120] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 , and C 12 , also includes any range from n to n+m, such as C 1-12 Including C 1-3 、C 1-6 、C 1-9 、C 3-6 、C 3-9 、C 3-12 、C 6-9 、C 6-12 , and C 9-12 etc.; similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring, and also includes any range from n to n+m, for example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 5-10-membered ring, a 6-7-membered ring, a 6-8-membered ring, a 6-9-membered ring and a 6-10-membered ring, etc.

[0121] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (e.g., an affine substitution reaction). For example, representative leaving groups include trifluoromethanesulfonate; chloro, bromo, iodo; sulfonate groups such as methanesulfonate, toluenesulfonate, p-bromobenzenesulfonate, p-toluenesulfonate, etc.; acyloxy groups such as acetoxy and trifluoroacetoxy, etc.

[0122] The term "protecting group" includes, but is not limited to, an "amino protecting group," a "hydroxy protecting group," or a "thiol protecting group." The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butyloxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-bis-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like. The term "hydroxy protecting group" refers to a protecting group suitable for preventing side reactions at the hydroxyl group. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and tert-butyl; acyl groups such as alkanoyl (e.g., acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like.

[0123] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

[0124] The solvent used in the present invention is commercially available.

[0125] Compounds are named according to the conventional nomenclature in the art or using Software naming, commercially available compounds use supplier catalog names. BRIEF DESCRIPTION OF THE DRAWINGS

[0126] Figure 1 1 is a graph showing the results of an LLC-PK1 cell proliferation inhibition experiment according to an embodiment of the present invention. DETAILED DESCRIPTION

[0127] The present application is described in detail below by way of examples, but this does not necessarily mean that there are any adverse limitations on the present application. The present application has been described in detail herein, and specific embodiments thereof have 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 application without departing from the spirit and scope of the present application.

[0128] Unless otherwise specified, the experimental materials and reagents used in the following examples can be obtained from commercial sources.

[0129] Preparation of intermediates

[0130] Reference Example 1: Preparation of Intermediate I-1

[0131]

[0132] At room temperature, p-toluenesulfonyl chloride (21.9 g, 115 mmol) was added to a solution of 7-chloro-1,2,3,4-tetrahydrobenzo[B]azepin-5-one (15 g, 76.7 mmol) in pyridine (150 mL). The reaction mixture was allowed to react at room temperature for 16 hours. The mixture was concentrated under reduced pressure, poured into water (200 mL), and extracted with ethyl acetate (100 mL x 3). The organic phases were combined. The organic phases were washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to remove the organic solvent, and the residue was purified by silica gel chromatography to obtain intermediate I-1.

[0133] LC-MS (ESI) [M+H] + 349.9.

[0134] 1 H NMR (400MHz, CDCl3) δ7.66(d,J=2.4Hz,1H),7.58(d,J=8.3Hz,2H),7.47(dd,J=8.6,2.5Hz,1H),7.43(d,J=8 .5Hz,1H),7.28(d,J=8.0Hz,2H),3.83(t,J=6.5Hz,2H),2.43(s,3H),2.40–2.35(m,2H),2.00–1.91(m,2H).

[0135] Reference Example 2: Preparation of Intermediate I-2

[0136]

[0137] At room temperature, intermediate I-1 (20.0 g, 57.2 mmol) was added to cyclohexane (250 mL), followed by n-butylamine (8.49 mL, 85.8 mmol) and trifluoroacetic acid (1.00 mL). The reaction mixture was refluxed under argon for 48 hours and concentrated under reduced pressure to obtain a residue. A mixture of ethyl acetate and petroleum ether (volume ratio 1:15, 64 mL) was added to the residue with stirring. After stirring for 10 minutes, the mixture was filtered and the resulting solid was dried under reduced pressure to obtain intermediate I-2.

[0138] 1 H NMR (400MHz, CDCl3) δ7.56–7.51(m,2H),7.45(d,J=2.2Hz,1H),7.41–7.38(m,1H),7.37–7.34( m,1H),7.22(d,J=8.0Hz,2H),3.76(t,J=6.2Hz,2H),2.93(t,J=7.2Hz,2H),2.40(s,3H),2.10– 2.06(m,2H),1.76–1.69(m,2H),1.65–1.58(m,2H),1.41-1.35(m,2H),0.96(t,J=7.4Hz,3H).

[0139] Reference Example 3: Preparation of Intermediate I-3

[0140]

[0141] At room temperature, a selective fluorine reagent (26.2 g, 74.0 mmol) was added to acetonitrile (200 mL). Intermediate I-2 (15.0 g, 37.0 mmol) was added in five batches, with one batch added every 40 minutes. After the addition was complete, the reaction mixture was stirred at room temperature for 72 hours. Ice water (200 mL) was added, and concentrated hydrochloric acid (15.0 mL) was slowly added dropwise with stirring. After stirring for five minutes, the mixture was filtered and the resulting solid was dried under reduced pressure to obtain Intermediate I-3.

[0142] 1 H NMR (400MHz, DMSO-d6) δ7.79–7.63(m,4H),7.44(d,J=8.0Hz,2H),7.37–7.32(m,1H),4.07–3.98(m,2H),2.67–2.53(m,2H),2.41(s,3H).

[0143] Reference Example 4: Preparation of Intermediate I-4

[0144]

[0145] Intermediate I-3 (12.0 g, 31.1 mmol) was added to concentrated sulfuric acid (15.0 mL) at 0°C, and the reaction mixture was stirred at room temperature for 4 hours. The mixture was poured into ice water, adjusted to pH 11 with 50% aqueous sodium hydroxide solution, and extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-4.

[0146] LCMS (ESI) [M+H] + 232.1.

[0147] 1 H NMR (400MHz, DMSO) δ7.45(dd,J=8.4,3.6Hz,2H),7.34(dd,J=8.9,2.6Hz,1H),6.91(d,J=8.9Hz,1H),3.29–3.21(m,2H),2.70–2.56(m,2H).

[0148] Reference Example 5: Preparation of Intermediate I-5

[0149]

[0150] Methyltriphenylphosphonium bromide (18.5 g, 51.8 mmol) was added to tetrahydrofuran (120 mL) at room temperature. The temperature was lowered to 0°C, and potassium tert-butoxide (5.81 g, 51.8 mmol) was added. The reaction mixture was stirred at 0°C for 1 hour, and then a solution of intermediate I-4 (6.00 g, 25.9 mmol) in tetrahydrofuran (20 mL) was added at 0°C. After the addition was complete, the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was poured into ice water (200 mL) and extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-5.

[0151] LCMS (ESI) [M+H] + 230.1.

[0152] Reference Example 6: Preparation of Intermediate I-6

[0153]

[0154] At room temperature, methyl 6-aminonicotinate (1.0 g, 6.57 mmol) was dissolved in pyridine (20 mL), and 2-trifluoromethylbenzoyl chloride (1.51 g, 7.25 mmol) was added. After the addition was complete, the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was poured into ice water (100 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with water (50 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography to obtain intermediate I-6.

[0155] LC-MS (ESI) [M+H] + 325.0.

[0156] Reference Example 7: Preparation of Intermediate I-7

[0157]

[0158] At room temperature, intermediate I-6 (1.35 g, 4.16 mmol) was dissolved in tetrahydrofuran (10 mL). A solution of sodium hydroxide (499 mg, 12.5 mmol) in water (2 mL) was added. After the addition was complete, the reaction mixture was stirred at 70°C for 1 hour. After completion of the reaction, the pH of the reaction solution was adjusted to 5-6 with 1N hydrochloric acid. The solid was filtered and dried to obtain intermediate I-7.

[0159] LC-MS (ESI) [M+H] + 311.0.

[0160] Reference Example 8: Preparation of Intermediate I-8

[0161]

[0162] Intermediate I-7 (244 mg, 0.785 mmol) was dissolved in N,N-dimethylacetamide (6.00 mL) at room temperature and cooled to 0°C. Thionyl chloride (125 mg, 1.05 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. A solution of Intermediate I-5 (120 mg, 0.523 mmol) in N,N-dimethylacetamide (3 mL) was then added, and the reaction mixture was stirred at room temperature for another 16 hours. The reaction mixture was added to water (30 mL) and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated sodium bicarbonate aqueous solution (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-8.

[0163] LCMS (ESI) [M+H] + 522.2.

[0164] Reference Example 9: Preparation of Intermediate I-9

[0165]

[0166] At room temperature, o-methylbenzoyl chloride (5.00 g, 32.3 mmol) was dissolved in chloroform (80 mL), and 4-amino-2-methylbenzoic acid (4.88 g, 32.3 mmol) and triethylamine (9.81 g, 96.9 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours. The reaction system was poured into water (200 mL) and extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-9.

[0167] 1 H NMR(400MHz, DMSO-d6)δ10.53(s,1H),7.87(d,J=8.4Hz,1H),7.73–7.62(m,2H),7 .51–7.45(m,1H),7.44–7.37(m,1H),7.36–7.25(m,2H),2.53(s,3H),2.39(s,3H).

[0168] Reference Example 10: Preparation of Intermediate I-10

[0169]

[0170] Intermediate I-9 (211 mg, 0.785 mmol) was dissolved in N,N-dimethylacetamide (6.00 mL) at room temperature and cooled to 0°C. Under argon, thionyl chloride (125 mg, 1.05 mmol) was added. The reaction mixture was stirred at room temperature for 3 hours, followed by a solution of Intermediate I-5 (120 mg, 0.523 mmol) in N,N-dimethylacetamide (3.00 mL). The reaction mixture was stirred at room temperature for another 16 hours. The reaction mixture was added to water (30 mL) and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed sequentially with saturated sodium bicarbonate aqueous solution (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-10.

[0171] LCMS (ESI) [M+H] + 481.1.

[0172] Reference Example 11: Preparation of Intermediate I-11

[0173]

[0174] Methyl 6-amino-4-methylnicotinate (500 mg, 3.01 mmol) was dissolved in pyridine (20 mL) at room temperature, and 2-trifluoromethylbenzoyl chloride (628 mg, 3.01 mmol) was added. After the addition was complete, the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was poured into ice water (100 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with water (50 mL × 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography to obtain intermediate I-11.

[0175] LC-MS (ESI) [M+H] + 339.1.

[0176] Reference Example 12: Preparation of Intermediate I-12

[0177]

[0178] Intermediate I-11 (700 mg, 2.07 mmol) was dissolved in tetrahydrofuran (10 mL) at room temperature, and a solution of sodium hydroxide (414 mg, 10.35 mmol) in water (5 mL) was added. After the addition was complete, the reaction mixture was stirred at 70°C for 1 hour. The pH of the reaction solution was adjusted to 5-6 with 1N hydrochloric acid, filtered, and the solid was dried to obtain Intermediate I-12.

[0179] Reference Example 13: Preparation of Intermediate I-13

[0180]

[0181] Intermediate I-12 (200 mg, 0.617 mmol) was dissolved in dichloromethane (10 mL) at 0°C. Oxalyl chloride (165 mg, 1.30 mmol) and N,N-dimethylformamide (1 drop) were added. After the addition was complete, the reaction mixture was stirred at 0°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the crude intermediate I-13. The crude product was used directly in the next reaction without further purification.

[0182] Reference Example 14: Preparation of Intermediate I-14

[0183]

[0184] 7-Chloro-1,2,3,4-tetrahydrobenzo[B]azepin-5-one (26.0 g, 0.133 mol) and di-tert-butyl dicarbonate (200 g) were heated to 100°C for 16 hours. After cooling to room temperature, most of the di-tert-butyl dicarbonate was removed under reduced pressure using an oil pump. The residue was separated and purified by silica gel chromatography to obtain intermediate I-14.

[0185] LC-MS (ESI) [M+H-56] + 239.9.

[0186] 1 H NMR (400MHz, DMSO-d6) δ7.66–7.58(m,2H),7.48(d,J=8.4Hz,1H),3.66(s,2H),2.64(t,J=6.6Hz,2H),2.07–1.96(m,2H),1.40(d,J=13.8Hz,9H).

[0187] Reference Example 15: Preparation of Intermediate I-15

[0188]

[0189] Under argon, methyltriphenylphosphonium bromide (47.9 g, 0.134 mol) was added to a solution of potassium tert-butoxide (18.9 g, 0.168 mol) in tetrahydrofuran (500 mL) at 0°C. The reaction mixture was stirred at 0°C for 30 minutes. Intermediate I-14 (33.0 g, 0.112 mol) was then added to the reaction system. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with water (500 mL) and extracted with ethyl acetate (500 mL x 2). The organic phases were combined, washed with water (500 mL) and saturated brine (500 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-15.

[0190] LC-MS (ESI) [M+H-56] + 238.0.

[0191] 1 H NMR(400MHz, DMSO-d6)δ7.36(s,1H),7.30(dd,J=8.5,2.5Hz,1H),7.22(d,J=8.5Hz,1H),5 .20(d,J=11.7Hz,2H),3.51(s,2H),2.37(t,J=6.1Hz,2H),1.81(s,2H),1.45–1.28(m,9H).

[0192] Reference Example 16: Preparation of Intermediate I-16

[0193]

[0194] Intermediate I-15 (6.10 g, 20.8 mmol) was dissolved in a mixture of trifluoroacetic acid and dichloromethane (20.0 mL / 40.0 mL) at room temperature, and the reaction mixture was stirred at room temperature for 3 hours. The mixture was poured into a saturated sodium bicarbonate solution (200 mL) and extracted with dichloromethane (50 mL x 4). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was separated and purified by silica gel chromatography to obtain intermediate I-16.

[0195] 1 H NMR(400MHz,DMSO-d6)δ7.11(d,J=2.5Hz,1H),7.00(dd,J=8.6,2.5Hz,1H),6.69(d,J=8.6Hz,1H), 5.17(d,J=1.6Hz,1H),4.99–4.93(m,1H),3.18–3.07(m,2H),2.50–2.45(m,2H),1.88–1.76(m,2H).

[0196] Reference Example 17: Preparation of Intermediate I-17

[0197]

[0198] At room temperature, intermediate I-13 (150 mg) was dissolved in pyridine (10 mL), and intermediate I-16 (60 mg, 0.310 mmol) was added. After the addition was complete, the reaction mixture was stirred at room temperature for 16 hours. The reaction solution was diluted with water (10 mL) and extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with water (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography to obtain intermediate I-17.

[0199] LC-MS (ESI) [M+H] + 500.1.

[0200] Reference Example 18: Preparation of Intermediate I-18

[0201]

[0202] At room temperature, o-methylbenzoyl chloride (6.11 g, 39.5 mmol) was added to a solution of methyl 6-aminonicotinate (5.00 g, 32.9 mmol) in pyridine (40.0 mL). The reaction mixture was stirred at room temperature for 2 hours, then poured into water (300 mL) and filtered. The resulting solid was dried to afford Intermediate I-18.

[0203] LC-MS (ESI) [M+H]+ 271.0.

[0204] Reference Example 19: Preparation of Intermediate I-19

[0205]

[0206] At room temperature, sodium hydroxide (2.66 g, 66.6 mmol) was added to a solution of intermediate I-18 (6.00 g, 22.2 mmol) in methanol / water (60 mL / 30 mL). The reaction mixture was stirred at 70°C for 30 minutes. After cooling to room temperature, the pH was adjusted to 5-6 with 3N dilute hydrochloric acid in an ice-water bath, and extracted with ethyl acetate (50 mL x 6). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain intermediate I-19.

[0207] LC-MS (ESI) [M+H] + 257.1.

[0208] Reference Example 20: Preparation of Intermediate I-20

[0209]

[0210] Intermediate I-19 (1.50 g, 5.85 mmol) and N,N-dimethylformamide (0.10 mL) were added to dichloromethane (20.0 mL) at room temperature. The reaction system was cooled to 0°C, and oxalyl chloride (1.49 g, 11.7 mmol) was added. The reaction solution was stirred at 0°C for 1 hour and concentrated to dryness to obtain the crude product, Intermediate I-20. The crude product was used directly in the next reaction without further purification.

[0211] Reference Example 21: Preparation of Intermediate I-21

[0212]

[0213] At room temperature, intermediate I-16 (378 mg, 1.95 mmol) was added to pyridine (5.00 mL), and intermediate I-20 (500 mg) was added all at once. The reaction mixture was stirred at room temperature for 16 hours, poured into water (30 mL), and extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-21.

[0214] LC-MS (ESI) [M+H] + 432.1.

[0215] Reference Example 22: Preparation of Intermediate I-22

[0216]

[0217] Intermediate I-7 (200 mg, 0.65 mmol) was dissolved in dichloromethane (10 mL) at 0°C. Oxalyl chloride (165 mg, 1.30 mmol) and N,N-dimethylformamide (1 drop) were added. After the addition was complete, the reaction mixture was stirred at 0°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the crude intermediate I-22. The crude product was used directly in the next reaction without further purification.

[0218] Reference Example 23: Preparation of Intermediate I-23

[0219]

[0220] At room temperature, intermediate I-22 (214 mg, 0.65 mmol) was dissolved in pyridine (10 mL), and intermediate I-16 (97 mg, 0.501 mmol) was added. After the addition was complete, the reaction mixture was stirred at room temperature for 16 hours. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with water (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel chromatography to obtain intermediate I-23.

[0221] LC-MS (ESI) [M+H] + 486.2.

[0222] Reference Example 24: Preparation of Intermediate I-24

[0223]

[0224] At room temperature, cuprous cyanide (24.0 g, 0.268 mol) was added to a solution of 2-amino-5-bromo-4-methylpyridine (25.0 g, 0.134 mol) in N,N-dimethylacetamide (230 mL). The reaction mixture was stirred at 170°C under argon for 16 hours. After cooling to 0°C, ethylenediamine (50 mL) and water (500 mL) were added to the reaction solution and stirred for 15 minutes to quench the reaction. The mixture was extracted with ethyl acetate (300 mL x 3). The organic phases were combined, washed with water (300 mL x 3) and saturated brine (500 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-24.

[0225] LC-MS (ESI) [M+H] + 134.1.

[0226] 1 H NMR (400MHz, DMSO-d6) δ8.23(s,1H),6.89(s,2H),6.35(s,1H),2.25(s,3H).

[0227] Reference Example 25: Preparation of Intermediate I-25

[0228]

[0229] Intermediate I-24 (10.2 g, 76.6 mmol) was added to a solution of sodium hydroxide (90 mL, 10 mol / L) and ethanol (90 mL) at room temperature. The reaction mixture was refluxed for 16 hours. After cooling to room temperature, the pH was adjusted to neutral with hydrochloric acid (6 mol / L). The precipitated solid was filtered, the filter cake washed with water, and dried to afford Intermediate I-25.

[0230] LC-MS (ESI) [M+H] + 153.3.

[0231] Reference Example 26: Preparation of Intermediate I-26

[0232]

[0233] At room temperature, thionyl chloride (15 mL) was added to a solution of intermediate I-25 (7.8 g, 51.3 mmol) in methanol (80 mL). The reaction mixture was refluxed for 16 hours. After cooling to room temperature, most of the methanol solvent was removed under reduced pressure, and the mixture was diluted with water (100 mL). The pH was adjusted to 13 with aqueous sodium hydroxide solution (2 mol / L). The precipitated solid was filtered, the filter cake was washed with water, and then dried to obtain intermediate I-26.

[0234] LC-MS (ESI) [M+H] + 167.1.

[0235] Reference Example 27: Preparation of Intermediate I-27

[0236]

[0237] o-Toluoyl chloride (5.14 g, 33.2 mmol) was added to a solution of intermediate I-26 (4.6 g, 27.7 mmol) in pyridine (35 mL) at room temperature. The reaction mixture was stirred at room temperature for 3 hours. Ice water (70 mL) was added to the reaction solution to precipitate a solid, which was filtered and the filter cake washed with plenty of water and petroleum ether and dried to obtain intermediate I-27.

[0238] LC-MS (ESI) [M+H] +285.2.

[0239] Reference Example 28: Preparation of Intermediate I-28

[0240]

[0241] At room temperature, 5% aqueous sodium hydroxide solution (48 mL) was added to a solution of intermediate I-27 (4.1 g, 14.4 mmol) in methanol (60 mL). The reaction mixture was reacted at 70°C for 30 minutes. After cooling to room temperature, the mixture was diluted with water (50 mL) and the pH was adjusted to weak acidity with hydrochloric acid (3 mol / L). The precipitated solid was filtered, and the filter cake was washed with plenty of water and lyophilized to obtain intermediate I-28.

[0242] LC-MS (ESI) [M+H] + 271.1.

[0243] 1 H NMR(400MHz,DMSO-d6)δ13.04(s,1H),10.99(s,1H),8.75(s,1H),8.16(s,1H),7.4 8(d,J=7.6Hz,1H),7.43–7.35(m,1H),7.32–7.24(m,2H),2.60(s,3H),2.39(s,3H).

[0244] Reference Example 29: Preparation of Intermediate I-29

[0245]

[0246] Oxalyl chloride (2.07 g, 16.3 mmol) and N,N-dimethylformamide (1 drop) were added to a solution of intermediate I-28 (2.2 g, 8.14 mmol) in dichloromethane (30 mL) under an ice-water bath. The reaction mixture was allowed to react at 0°C for 30 minutes. The solvent was concentrated to dryness at low temperature to obtain the crude intermediate I-29, which was used directly in the next reaction without purification.

[0247] Reference Example 30: Preparation of Intermediate I-30

[0248]

[0249] Intermediate I-29 (1.75 g) was added to a solution of Intermediate I-16 (900 mg, 4.65 mmol) in pyridine (30 mL) at room temperature. The reaction mixture was stirred at room temperature for 3 hours. Water (50 mL) was added and extracted with ethyl acetate (30 mL × 2). The organic phases were combined and washed with water (30 mL × 3) and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-30.

[0250] LC-MS (ESI) [M+H] + 446.2.

[0251] Reference Example 31: Preparation of Intermediate I-31

[0252]

[0253] N-Bromosuccinimide (110 mg, 0.618 mmol) was added to a solution of Intermediate I-30 (230 mg, 0.516 mmol) in tetrahydrofuran / water (15 mL / 3 mL) at room temperature, and the reaction was stirred at room temperature for 16 hours. Aqueous sodium hydroxide (2 mL, 2.5 N) was then added, and the reaction was stirred at room temperature for 2 hours. The reaction was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain the crude product. The crude product was purified by silica gel chromatography to obtain Intermediate I-31.

[0254] LC-MS (ESI) [M+H] + 462.0.

[0255] 1 H NMR (400MHz, DMSO-d6) δ10.97–10.53(m,1H),8.47–7.63(m,2H),7.56–7.10(m,6H),7.03–6.66(m,1 H),4.98–4.67(m,1H),3.30–2.62(m,3H),2.48–2.25(m,6H),2.21–1.99(m,2H),1.91–1.52(m,2H).

[0256] Reference Example 32: Preparation of Intermediate I-32

[0257]

[0258] 2-Methyl-4-nitrobenzoyl chloride (2.06 g, 10.3 mmol) and triethylamine (3.13 g, 30.9 mmol) were added to a solution of Intermediate I-16 (2.0 g, 10.3 mmol) in dichloromethane (40 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour, poured into water (100 mL), and extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-32.

[0259] LC-MS (ESI) [M+H] + 356.9.

[0260] Reference Example 33: Preparation of Intermediate I-33

[0261]

[0262] Iron powder (282 mg, 5.05 mmol) and ammonium chloride (540 mg, 10.1 mmol) were added to a solution of intermediate I-32 (360 mg, 1.01 mmol) in methanol / water (20 mL / 5 mL) at room temperature. Under argon, the reaction mixture was stirred at 60°C for 3 hours. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated to dryness to obtain the crude product. The crude product was purified by silica gel chromatography to obtain intermediate I-33.

[0263] LC-MS (ESI) [M+H] + 327.0.

[0264] Reference Example 34: Preparation of Intermediate I-34

[0265]

[0266] At room temperature, o-chlorobenzoyl chloride (225 mg, 1.29 mmol) and triethylamine (260 mg, 2.59 mmol) were added to a solution of intermediate I-33 (280 mg, 0.857 mmol) in 1,2-dichloroethane (20 mL). The reaction mixture was stirred at 50°C for 2 hours. After cooling to room temperature, the reaction solution was diluted with dichloromethane (20 mL), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-34.

[0267] LC-MS (ESI) [M+H] + 465.2.

[0268] Reference Example 35: Preparation of Intermediate I-35

[0269]

[0270] At room temperature, intermediate I-9 (2.00 g, 7.43 mmol) was added to a solution of thionyl chloride (20.0 mL), and the reaction mixture was stirred at 40°C for 4 hours. The reaction system was concentrated under reduced pressure to obtain the crude intermediate I-35, which was used directly in the next reaction.

[0271] Reference Example 36: Preparation of Intermediate I-36

[0272]

[0273] At room temperature, intermediate I-16 (1.42 g, 7.31 mmol) was dissolved in pyridine (30.0 mL), and intermediate I-35 (2.00 g) was added. The reaction mixture was stirred at room temperature for 8 hours. The reaction system was poured into water (100 mL) and extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-36.

[0274] LC-MS (ESI) [M+H] + 445.0.

[0275] Reference Example 37: Preparation of Intermediate I-37

[0276]

[0277] Intermediate I-36 (1.00 g, 2.25 mmol) was dissolved in dichloromethane (30.0 mL) at room temperature, and m-chloroperbenzoic acid (1.95 g, 11.3 mmol) was added. The reaction system was stirred at room temperature for 10 hours. The system was concentrated under reduced pressure at room temperature to obtain a crude product, which was then separated and purified by C18 reverse-phase chromatography to obtain intermediate I-37.

[0278] 1 H NMR (400MHz, DMSO-d6) δ10.41–10.23(m,1H),7.77–7.10(m,9H),6.83–6.63(m,1 H),5.04–3.53(m,1H),3.21–2.68(m,3H),2.37–2.33(m,6H),2.13–1.56(m,4H).

[0279] Reference Example 38: Preparation of Intermediate I-38

[0280]

[0281] At 25°C, intermediate I-1 (1.00 g, 2.86 mmol) was dissolved in tetrahydrofuran (10 mL). Tert-butylsulfenamide (415.74 mg, 3.43 mmol) and tetraethyl titanate (1.30 g, 5.72 mmol) were added and stirred in a microwave at 80°C for 3 hours. Water (10 mL) was added, the mixture was filtered, and the filter cake was washed with ethyl acetate (20 mL). The filtrate was collected and washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-38.

[0282] LC-MS (ESI) [2M+H] + 453.0.

[0283] Reference Example 39: Preparation of Intermediate I-39

[0284]

[0285] Zinc powder (2.01 g, 30.76 mmol) was suspended in tetrahydrofuran (100 mL) at 25°C and the atmosphere was replaced with nitrogen three times. 1,2-Dibromoethane (96.31 mg, 0.51 mmol) and trimethylsilyl chloride (278.48 mg, 2.56 mmol) were added and stirred at 65°C for 1 hour. Tert-butyl bromoacetate (5.00 g, 25.63 mmol) was added dropwise. After the addition was complete, the mixture was stirred at 50°C for 1 hour. Cooling gave a tetrahydrofuran solution of Intermediate I-39 (100 mL, 25.63 mmol).

[0286] Reference Example 40: Preparation of Intermediate I-40

[0287]

[0288] To a solution of Intermediate I-39 in tetrahydrofuran (100 mL, 25.63 mmol) was added dropwise a solution of Intermediate I-38 (850.00 mg, 1.88 mmol) in tetrahydrofuran (20 mL) at 25°C. The mixture was stirred at 50°C for 16 hours. Water (100 mL) was added and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-40.

[0289] LC-MS (ESI) [M+H] + 569.2.

[0290] Reference Example 41: Preparation of Intermediate I-41

[0291]

[0292] At 25°C, intermediate I-40 (350.00 mg, 0.61 mmol) was dissolved in tetrahydrofuran (10 mL). A solution of diisobutylaluminum hydride in toluene (2.05 mL, 3.07 mmol, 1.5 M) was added and allowed to react at room temperature for 16 hours. Water (0.12 mL), 15% aqueous sodium hydroxide solution (0.12 mL), and water (0.3 mL) were added in sequence, followed by anhydrous sodium sulfate. The mixture was stirred at room temperature for 0.5 hours. The mixture was filtered, the filtrate was concentrated, and the mixture was purified by silica gel chromatography to afford intermediate I-41.

[0293] LC-MS (ESI) [M+H] + 499.2.

[0294] Reference Example 42: Preparation of Intermediate I-42

[0295]

[0296] At 25°C, intermediate I-41 (170.00 mg, 0.34 mmol) was dissolved in toluene (10 mL). Cyanomethylidenetri-n-butylphosphine (98.47 mg, 0.41 mmol) was added and stirred at 110°C for 3 hours. After cooling to room temperature, water (10 mL) was added and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The mixture was purified by silica gel chromatography to afford intermediate I-42.

[0297] LC-MS (ESI) [M+H] + 481.2.

[0298] Reference Example 43: Preparation of Intermediate I-43

[0299]

[0300] Intermediate I-42 (100.00 mg, 0.21 mmol) was dissolved in concentrated hydrochloric acid (5 mL) at 25°C and reacted at 100°C for 16 hours. The mixture was directly concentrated to dryness and purified by C18 reverse phase column to obtain intermediate I-43.

[0301] LC-MS (ESI) [M+H] + 223.0.

[0302] Reference Example 44: Preparation of Intermediate I-44

[0303]

[0304] Intermediate I-43 (40.00 mg, 0.18 mmol) was dissolved in tetrahydrofuran (5 mL) at 25°C. 10% aqueous sodium bicarbonate (5 mL) and di-tert-butyl dicarbonate (47.04 mg, 0.22 mmol) were added and allowed to react at room temperature for 1 hour. Water (10 mL) was added and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The mixture was then purified by silica gel chromatography to afford Intermediate I-44.

[0305] LC-MS (ESI) [M+H] + 323.2.

[0306] Reference Example 45: Preparation of Intermediate I-45

[0307]

[0308] At 25°C, intermediate I-44 (50.00 mg, 0.15 mmol) was dissolved in dichloromethane (10 mL). Triethylamine (47.02 mg, 0.46 mmol) and 2-methyl-4-nitrobenzoyl chloride (46.37 mg, 0.23 mmol) were added and stirred at room temperature for 16 hours. Water (10 mL) was added and the mixture was extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The mixture was then purified by silica gel chromatography to afford intermediate I-45.

[0309] LC-MS (ESI) [M-100+H] + 386.2.

[0310] Reference Example 46: Preparation of Intermediate I-46

[0311]

[0312] Intermediate I-45 (50.00 mg, 0.10 mmol) was dissolved in ethanol (4 mL) at 25°C. Reduced iron powder (28.73 mg, 0.51 mmol) and saturated aqueous ammonium chloride (2 mL) were added and stirred at 80°C for 5 hours. The mixture was filtered, and the filter cake was washed with ethyl acetate (10 mL). The filtrate was collected and separated by adding water (10 mL). The organic phase was collected, concentrated, and purified by silica gel chromatography to obtain Intermediate I-46.

[0313] LC-MS (ESI) [M-100+H] + 356.2.

[0314] Reference Example 47: Preparation of Intermediate I-47

[0315]

[0316] Intermediate I-46 (30.00 mg, 0.066 mmol) was dissolved in dichloromethane (5 mL) at 25°C. Triethylamine (19.97 mg, 0.20 mmol) and 2-methylbenzoyl chloride (12.21 mg, 0.079 mmol) were added sequentially and allowed to react at room temperature for 1 hour. Water (10 mL) was added and the mixture was extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. The mixture was then purified by silica gel chromatography to afford Intermediate I-47.

[0317] LC-MS (ESI) [M-100+H] + 474.2.

[0318] Reference Example 48: Preparation of Intermediate I-48

[0319]

[0320] At room temperature, cesium carbonate (82.7 g, 254 mmol) and palladium tetrakistriphenylphosphine (2.93 g, 2.54 mmol) were added to a solution of 2-bromo-4-chloro-1-nitrobenzene (20.0 g, 84.6 mmol) and 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (21.3 g, 101.4 mmol) in dioxane / water (600 mL / 200 mL). The reaction mixture was stirred at 100°C under argon for 5 hours. After cooling to room temperature, the solvent was mostly removed under reduced pressure, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain the crude product. The crude product was purified by silica gel chromatography to obtain intermediate I-48.

[0321] LC-MS (ESI) [M+H] + 240.0.

[0322] 1 H NMR (400MHz, CD3OD) δ7.92 (d, J=8.7Hz, 1H), 7.52 (dd, J=8.7, 2.3Hz, 1H), 7.44 (d, J=2.3 Hz,1H),5.75–5.71(m,1H),4.26–4.21(m,2H),3.89(t,J=5.3Hz,2H),2.36–2.31(m,2H).

[0323] Reference Example 49: Preparation of Intermediate I-49

[0324]

[0325] At room temperature, m-chloroperbenzoic acid (16.2 g, 93.9 mmol) was added to a solution of Intermediate I-48 (15.0 g, 62.6 mmol) in dichloromethane (100 mL). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with dichloromethane (100 mL) and washed with saturated aqueous sodium thiosulfate (100 mL × 3), saturated aqueous sodium carbonate (100 mL × 3), and saturated brine (200 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-49.

[0326] LC-MS (ESI) [M+H] + 256.0.

[0327] 1 H NMR(400MHz, DMSO-d6)δ8.17(d,J=8.7Hz,1H),7.77–7.68(m,2H),4.01–3.88(m,2H),3. 69–3.61(m,1H),3.53–3.45(m,1H),3.37(s,1H),2.20–2.11(m,1H),2.03–1.95(m,1H).

[0328] Reference Example 50: Preparation of Intermediate I-50

[0329]

[0330] Boron trifluoride etherate (4.37 g, 30.8 mmol) was added to a solution of intermediate I-49 (7.5 g, 29.3 mmol) in dichloromethane (70 mL) under an ice-water bath. The reaction mixture was stirred at 0°C for 30 minutes. The reaction mixture was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-50.

[0331] LC-MS (ESI) [M+H] + 256.0.

[0332] 1H NMR (400MHz, DMSO-d6) δ9.56 (s, 1H), 8.07 (d, J = 8.7Hz, 1H), 7.72 (dd, J = 8.7, 2.2Hz, 1H), 7.65 (d, J = 3.6Hz, 1H), 4.31 (d, J=9.8Hz, 1H), 4.05 (d, J=9.9Hz, 1H), 4.00–3.86 (m, 2H), 2.69–2.60 (m, 1H), 2.40–2.31 (m, 1H).

[0333] Reference Example 51: Preparation of Intermediate I-51

[0334]

[0335] Under argon protection in an ice-water bath, sodium hydride (278 mg, 60% wt, 6.95 mmol) was added to triethylphosphonoacetate (1.18 g, 5.26 mmol) in tetrahydrofuran (30 mL). The mixture was stirred in an ice-water bath for 30 minutes, and then intermediate I-50 (890 mg, 3.48 mmol) was added. The reaction mixture was reacted at room temperature for 2 hours. Water (30 mL) was added to the reaction mixture and extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-51.

[0336] LC-MS (ESI) [M+H] + 326.1.

[0337] 1 H NMR (400MHz, DMSO-d6) δ7.91–7.84(m,1H),7.70–7.63(m,2H),7.08(d,J=15.9Hz,1H),5.68(d,J=15.9Hz ,1H),4.18–4.06(m,3H),3.92–3.81(m,3H),2.48–2.43(m,1H),2.36–2.27(m,1H),1.19(t,J=7.1Hz,3H).

[0338] Reference Example 52: Preparation of Intermediate I-52

[0339]

[0340] Platinum dioxide (50 mg) was added to a solution of intermediate I-51 (720 mg, 2.21 mmol) in tetrahydrofuran (50 mL) at room temperature. The reaction mixture was hydrogenated using a hydrogen balloon at room temperature for approximately 6 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was purified by silica gel chromatography to obtain intermediate I-52.

[0341] LC-MS (ESI) [M+H] + 298.1.

[0342] Reference Example 53: Preparation of Intermediate I-53

[0343]

[0344] Sodium hydroxide (88.7 mg, 2.22 mmol) was added to a solution of intermediate I-52 (220 mg, 0.739 mmol) in tetrahydrofuran / water (10 mL / 3 mL) at room temperature. The reaction mixture was stirred at 40°C for 16 hours. After cooling to room temperature, the reaction mixture was adjusted to pH 7 with dilute hydrochloric acid (1N) and concentrated under reduced pressure to remove the organic solvent to afford crude intermediate I-53, which was used directly in the next step without purification.

[0345] LC-MS (ESI) [M+H] + 270.1.

[0346] Reference Example 54: Preparation of Intermediate I-54

[0347]

[0348] 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (283 mg, 1.48 mmol) and 4-dimethylaminopyridine (135 mg, 1.11 mmol) were added to a solution of Intermediate I-53 (300 mg) in tetrahydrofuran (60 mL) at room temperature. The reaction mixture was allowed to react at room temperature for 16 hours. Water (60 mL) was added to the reaction mixture and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-54.

[0349] LC-MS (ESI) [M+H] + 252.1.

[0350] 1H NMR (400MHz, DMSO-d6) δ9.57(d,J=30.6Hz,1H),7.40–7.29(m,2H),7.04–6.97(m,1H),3.90–3.78(m,3H),3.73–3.65(m,1H),2.31–2.05(m,6H).

[0351] Reference Example 55: Preparation of Intermediate I-55

[0352]

[0353] At room temperature, a borane-tetrahydrofuran solution (2.09 mL, 1 M) was added to a solution of intermediate I-54 (105 mg, 0.417 mmol) in tetrahydrofuran (20 mL). The reaction mixture was allowed to react at 40°C for 2 hours. After cooling to room temperature, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was purified by silica gel chromatography to obtain intermediate I-55.

[0354] LC-MS (ESI) [M+H] + 238.1.

[0355] 1 H NMR (400MHz, CDCl3) δ7.21(d,J=2.3Hz,1H),7.00(dd,J=8.3,2.4Hz,1H),6.65(d,J=8.3Hz,1H),4.10(d,J=8.8Hz,1H),3.97–3.89(m,2H ),3.81–3.74(m,1H),3.14–3.05(m,1H),3.00–2.93(m,1H),2.40–2.32(m,1H),2.18–2.09(m,1H),1.89–1.83(m,2H),1.75–1.69(m,2H).

[0356] Reference Example 56: Preparation of Intermediate I-56

[0357]

[0358] At room temperature, 2-methyl-4-nitrobenzoyl chloride (42.4 mg, 0.212 mmol) was added to a solution of intermediate I-55 (42 mg, 0.177 mmol) and pyridine (42.0 mg, 0.531 mmol) in tetrahydrofuran (5 mL). The reaction mixture was reacted at 50°C for 5 hours. After cooling to room temperature, the mixture was diluted with water (15 mL) and extracted with ethyl acetate (10 mL x 2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-56.

[0359] LC-MS (ESI) [M+H] + 401.0.

[0360] Reference Example 57: Preparation of Intermediate I-57

[0361]

[0362] Iron powder (31.3 mg, 0.560 mmol) and ammonium chloride (59.9 mg, 1.12 mmol) were added to a solution of intermediate I-56 (45 mg, 0.112 mmol) in methanol / water (10 mL / 3 mL) at room temperature. Under argon protection, the reaction solution was reacted at 50°C for 2 hours. After cooling to room temperature, filtering, the filtrate was concentrated to remove most of the solvent, diluted with water (5 mL), and extracted with ethyl acetate (10 mL × 3). The organic phases were combined and washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-57.

[0363] LC-MS (ESI) [M+H] + 371.0.

[0364] Reference Example 58: Preparation of Intermediate I-58

[0365]

[0366] Trimethylsulfoxide iodide (370 mg, 1.68 mmol) and potassium tert-butoxide (189 mg, 1.68 mmol) were added to dimethylsulfoxide (6.00 mL) at room temperature. The mixture was stirred under argon for 30 minutes. Intermediate I-4 (130 mg, 0.561 mmol) was then added at room temperature. The reaction mixture was stirred at room temperature for another 3 hours. Water (20 mL) was added and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-58.

[0367] LCMS (ESI) [M+H] + 260.0.

[0368] 1 HNMR (400MHz, DMSO-d6) δ7.57(d,J=2.6Hz,1H),7.17(dd,J=8.4,2.6Hz,1H),6.86(d,J=8.4Hz,1H),5.65(d,J=4 .9Hz,1H),4.57–4.40(m,2H),3.22–3.05(m,2H),2.73(t,J=12.4Hz,1H),2.41–2.32(m,1H),2.20–2.04(m,1H).

[0369] Reference Example 59: Preparation of Intermediate I-59

[0370]

[0371] N-Bromosuccinimide (395 mg, 2.22 mmol) was added to a solution of Intermediate I-21 (800 mg, 1.85 mmol) in tetrahydrofuran / water (15 mL / 3 mL) at room temperature, and the reaction mixture was stirred at room temperature for 24 hours. A 10% aqueous sodium hydroxide solution (3.70 mL, 9.25 mmol) was then added. The reaction mixture was stirred at room temperature for another hour. The reaction mixture was extracted with ethyl acetate (15 mL x 3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified on a C18 reverse-phase column to obtain Intermediate I-59.

[0372] LC-MS (ESI) [M+H] + 448.1.

[0373] 1 H NMR (400MHz, DMSO-d6) δ10.87(m,2H),8.20–6.90(m,10H),4.94–4.81(m,1H),3.13–2.78(m,3H),2.35(s,3H),2.15–1.93(m,2H),1.66(m,2H).

[0374] Reference Example 60: Preparation of Intermediate I-60

[0375]

[0376] 2-Phenylbenzoic acid (14.0 g, 70.6 mmol) was dissolved in dichloromethane (200 mL) at room temperature. Thionyl chloride (16.8 g, 141 mmol) was added to the mixture, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure to obtain crude intermediate I-60, which was used directly in the next reaction.

[0377] Reference Example 61: Preparation of Intermediate I-61

[0378]

[0379] At room temperature, intermediate I-60 (10.0 g) was dissolved in pyridine (80.0 mL), and methyl 6-aminonicotinate (7.03 g, 46.2 mmol) was added. The reaction mixture was stirred at room temperature for 3 hours. The reaction system was poured into water (500 mL) and extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified on a C18 reverse-phase column to obtain intermediate I-61.

[0380] LC-MS (ESI) [M+H] + 333.0.

[0381] Reference Example 62: Preparation of Intermediate I-62

[0382]

[0383] Intermediate I-61 (10.0 g, 30.1 mmol) was dissolved in methanol / water (100 mL / 20 mL) at room temperature. Sodium hydroxide (3.61 g, 90.3 mmol) was added to the mixture, and the reaction mixture was stirred at room temperature for 3 hours. The pH of the mixture was adjusted to 6-7 with dilute hydrochloric acid (1.0 mol / L). A large amount of solid precipitated, which was collected by filtration and dried under reduced pressure to obtain Intermediate I-62.

[0384] LC-MS (ESI) [M+H] + 319.0.

[0385] Reference Example 63: Preparation of Intermediate I-63

[0386]

[0387] Intermediate I-62 (2.00 g, 6.28 mmol) was dissolved in dichloromethane (30.0 mL) at room temperature, and thionyl chloride (2.24 g, 18.8 mmol) was added. The reaction mixture was stirred at 50°C for 3 hours. The reaction system was concentrated under reduced pressure to obtain crude intermediate I-63, which was used directly in the next reaction.

[0388] Reference Example 64: Preparation of Intermediate I-64

[0389]

[0390] At room temperature, intermediate I-16 (633 mg, 3.27 mmol) was dissolved in pyridine (10.0 mL), and intermediate I-63 (1.10 g) was added. The reaction mixture was stirred at room temperature for 3 hours. The system was poured into water (50.0 mL) and extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-64.

[0391] LC-MS (ESI) [M+H] + 494.2.

[0392] Reference Example 65: Preparation of Intermediate I-65

[0393]

[0394] Intermediate I-64 (500 mg, 1.01 mmol) was dissolved in tetrahydrofuran / water (25 mL / 5 mL) at room temperature. N-bromosuccinimide (360 mg, 2.02 mmol) was added, and the reaction mixture was stirred at room temperature for 24 hours. A 25% aqueous sodium hydroxide solution (1.00 mL) was added to the reaction system, and the system was stirred at room temperature for 1 hour. The reaction system was poured into water (20 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-65.

[0395] LC-MS (ESI) [M+H] + 510.1.

[0396] Reference Example 66: Preparation of Intermediate I-66

[0397]

[0398] In an ice-water bath, dissolve N-methylurea (5.0 g, 67.5 mmol) in water (40 mL), then add sodium nitrite (5.12 g, 74.2 mmol). Slowly add concentrated hydrochloric acid (8.9 mL) dropwise over approximately 30 minutes while stirring in an ice-water bath. Allow to react in an ice-water bath for 30 minutes. Filter the precipitate, wash the filter cake with water, and pump dry to obtain intermediate I-66, which is used directly in the next reaction without further purification.

[0399] Reference Example 67: Preparation of Intermediate I-67

[0400]

[0401] Under an ice-water bath, add 40% aqueous potassium hydroxide solution (40 mL) and diethyl ether (54 mL) to a smooth reaction flask. Add intermediate I-66 (4.2 g, 40.7 mmol) in batches. Gently shake the flask several times. After 30 minutes of reaction, transfer the upper ether layer to another smooth flask containing solid potassium hydroxide and dry. The resulting diethyl ether solution (0.75 M) of intermediate I-67 is used directly in the next reaction.

[0402] Reference Example 68: Preparation of Intermediate I-68

[0403]

[0404] Under an ice-water bath, a solution of intermediate I-67 in diethyl ether (40 mL, 0.75 M) was slowly added to a solution of intermediate I-15 (300 mg, 1.02 mmol) and palladium acetate (50 mg) in diethyl ether (10 mL). The mixture was stirred under ice-water bath for 1 hour. The reaction mixture was quenched with acetic acid (1 mL), diluted with water (30 mL), and extracted with diethyl ether (20 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to remove the organic solvent to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-68.

[0405] LC-MS (ESI) [M+H-56] + 252.1.

[0406] Reference Example 69: Preparation of Intermediate I-69

[0407]

[0408] Trifluoroacetic acid (1 mL) was added to a solution of intermediate I-68 (90 mg, 0.292 mmol) in dichloromethane (5 mL) at room temperature. The reaction mixture was stirred at room temperature for 5 hours. The mixture was concentrated under reduced pressure to dryness to obtain the crude intermediate I-69, which was used directly in the next reaction.

[0409] LC-MS (ESI) [M+H] + 208.1.

[0410] Reference Example 70: Preparation of Intermediate I-70

[0411]

[0412] 2-Methyl-4-nitrobenzoyl chloride (115 mg, 0.576 mmol) was added to a solution of intermediate I-69 (60 mg) and pyridine (68.6 mg, 0.867 mmol) in 1,2-dichloroethane (10 mL) at room temperature. The reaction mixture was stirred at 50°C for 16 hours. After cooling to room temperature, the mixture was diluted with dichloromethane (20 mL), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-70.

[0413] LC-MS (ESI) [M+H] + 371.1.

[0414] Reference Example 71: Preparation of Intermediate I-71

[0415]

[0416] Iron powder (30.2 mg, 0.541 mmol) and ammonium chloride (57.8 mg, 1.08 mmol) were added to a solution of intermediate I-70 (40 mg, 0.108 mmol) in methanol / water (10 mL / 3 mL) at room temperature. Under argon, the reaction mixture was stirred at 50°C for 3 hours. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated to dryness to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-71.

[0417] LC-MS (ESI) [M+H] + 341.1.

[0418] Reference Example 72: Preparation of Intermediate I-72

[0419]

[0420] At room temperature, diethyl malonate (10.0 g, 62.5 mmol) was dissolved in dimethyl sulfoxide (150 mL). Sodium hydroxide (2.92 g, 60% wt, 72.9 mmol) was slowly added. The reaction mixture was stirred at room temperature for 30 minutes. 2,4-Dichloronitrobenzene (10.0 g, 52.1 mmol) was then added to the reaction mixture. After the addition was complete, the mixture was heated to 80°C and stirred for 5 hours. The mixture was cooled to room temperature, water (300 mL) was added, and the mixture was extracted with ethyl acetate (300 mL x 3). The organic phases were combined, washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain the crude product. The crude product was purified by silica gel chromatography to obtain intermediate I-72.

[0421] LC-MS (ESI) [M+H] + 316.0.

[0422] Reference Example 73: Preparation of Intermediate I-73

[0423]

[0424] At room temperature, intermediate I-72 (10.0 g, 31.68 mmol) was dissolved in dimethyl sulfoxide (100 mL) and water (1 mL). Lithium chloride (6.65 g, 158.4 mmol) was added. After the addition was complete, the mixture was heated to 100°C and stirred for 5 hours. After cooling to room temperature, the mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL x 3). The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-73.

[0425] 1 H NMR (400MHz, CDCl3) δ8.08(d,J=8.8Hz,1H),7.44(dd,J=8.8,2.3Hz,1H),7.35(d,J=2.2Hz,1H),4.17(q,J=7.1Hz,2H),3.99(s,2H),1.25(t,J=7.1Hz,3H).

[0426] Reference Example 74: Preparation of Intermediate I-74

[0427]

[0428] At 0°C, intermediate I-73 (3.5 g, 14.4 mmol) was dissolved in N,N-dimethylformamide (100 mL) and slowly added to a solution of sodium hydroxide (860 mg, 60% wt, 21.5 mmol) in N,N-dimethylformamide (10 mL). The reaction mixture was stirred for 30 minutes. 1,3-Diiodopropane (5.1 g, 17.2 mmol) was added to the reaction mixture. After the addition was complete, the mixture was stirred at room temperature overnight. The mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL x 3). The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain the crude product. The crude product was purified by silica gel chromatography to obtain intermediate I-74.

[0429] 1H NMR (400MHz, CDCl3) δ7.88(d,J=8.6Hz,1H),7.44(d,J=2.2Hz,1H),7.37(dd,J=8.6,2.2Hz,1H),4.2 0(q,J=7.1Hz,2H),2.85–2.76(m,2H),2.46–2.31(m,3H),1.92–1.80(m,1H),1.22(t,J=7.1Hz,3H).

[0430] Reference Example 75: Preparation of Intermediate I-75

[0431]

[0432] At room temperature, intermediate I-74 (1.3 g, 4.58 mmol) was dissolved in methanol (10 mL) and water (10 mL). Sodium hydroxide (916 mg, 22.9 mmol) was added and the mixture was heated to 100°C with stirring for 3 hours. After cooling to room temperature, the reaction solution was concentrated to remove most of the methanol. The solution was then acidified with 1N hydrochloric acid and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain intermediate I-75.

[0433] 1 H NMR(400MHz, DMSO-d6)δ12.52(s,1H),7.97(d,J=8.6Hz,1H),7.65–7.55(m,2H), 2.67–2.58(m,2H),2.39(m,2H),2.17(dt,J=19.4,8.9Hz,1H),1.85–1.72(m,1H).

[0434] Reference Example 76: Preparation of Intermediate I-76

[0435]

[0436] At room temperature, a borane-tetrahydrofuran solution (19.5 mL, 1 M) was added to a solution of intermediate I-75 (1.0 g, 3.91 mmol) in tetrahydrofuran (50 mL). The reaction mixture was stirred at room temperature overnight. Water (50 mL) was added for dilution, and the mixture was extracted with ethyl acetate (50 mL x 2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was purified by silica gel chromatography to obtain intermediate I-76.

[0437] Reference Example 77: Preparation of Intermediate I-77

[0438]

[0439] At room temperature, intermediate I-76 (800 mg, 3.31 mmol) was dissolved in dichloromethane (10 mL). Dess-Martin periodinane (4.21 g, 9.93 mmol) was added. After the addition was complete, the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered, the filtrate was diluted with water (20 mL), and extracted with dichloromethane (20 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-77.

[0440] Reference Example 78: Preparation of Intermediate I-78

[0441]

[0442] At 0°C, sodium hydride (157 mg, 60% wt, 3.92 mmol) was added to triethylphosphonoacetate (659 mg, 2.94 mmol) in tetrahydrofuran (30 mL) under argon protection. The mixture was stirred in an ice-water bath for 30 minutes, and then intermediate I-77 (470 mg, 1.96 mmol) was added. The reaction mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated to remove most of the tetrahydrofuran, water (50 mL) was added, and the mixture was extracted with ethyl acetate (50 mL x 3). The aqueous phase was acidified with 1N hydrochloric acid and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain intermediate I-78.

[0443] 1 H NMR (400MHz, DMSO-d6) δ12.22(s,1H),7.88(d,J=8.7Hz,1H),7.59(dd,J=8.7,2.3Hz,1H),7.47(d,J=2.3Hz,1H),7. 30(d,J=15.7Hz,1H),5.65(d,J=15.7Hz,1H),2.43–2.33(m,4H),2.05(dd,J=20.5,10.1Hz,1H),1.79–1.70(m,1H).

[0444] Reference Example 79: Preparation of Intermediate I-79

[0445]

[0446] Intermediate I-78 (360 mg, 1.28 mmol) was dissolved in tetrahydrofuran (20 mL) at room temperature, and Raney nickel (300 mg) was added. After the addition was complete, the reaction mixture was stirred under a hydrogen atmosphere for 16 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain the crude intermediate I-79. The crude product was used directly in the next reaction.

[0447] LC-MS (ESI) [M+H] + 254.3.

[0448] Reference Example 80: Preparation of Intermediate I-80

[0449]

[0450] 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (331 mg, 1.73 mmol) and 4-dimethylaminopyridine (211 mg, 1.73 mmol) were added to a solution of Intermediate I-79 (220 mg) in tetrahydrofuran (60 mL) at room temperature, and the reaction mixture was stirred at room temperature for 16 hours. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-80.

[0451] LC-MS (ESI) [M+H] + 236.2.

[0452] Reference Example 81: Preparation of Intermediate I-81

[0453]

[0454] At room temperature, a borane-tetrahydrofuran solution (1.9 mL, 1 M) was added to a solution of intermediate I-80 (90 mg, 0.382 mmol) in tetrahydrofuran (20 mL). The reaction mixture was allowed to react at 40°C for 2 hours. After cooling to room temperature, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-81.

[0455] LC-MS (ESI) [M+H] + 222.2.

[0456] Reference Example 82: Preparation of Intermediate I-82

[0457]

[0458] At room temperature, 2-methyl-4-nitrobenzoyl chloride (94.8 mg, 0.475 mmol) was added to a solution of intermediate I-81 (70 mg, 0.316 mmol) and pyridine (75.0 mg, 0.948 mmol) in tetrahydrofuran (5 mL). The reaction mixture was stirred at 50°C for 5 hours. After cooling to room temperature, the mixture was diluted with water (15 mL) and extracted with ethyl acetate (10 mL x 2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-82.

[0459] LC-MS (ESI) [M+H] + 385.2.

[0460] Reference Example 83: Preparation of Intermediate I-83

[0461]

[0462] Iron powder (39.2 mg, 0.700 mmol) and ammonium chloride (74.9 mg, 1.40 mmol) were added to a solution of intermediate I-82 (54 mg, 0.140 mmol) in methanol / water (10 mL / 3 mL) at room temperature. Under argon protection, the reaction solution was stirred at 50°C for 2 hours. After cooling to room temperature, the mixture was filtered, and the filtrate was concentrated to remove most of the solvent. Water (5 mL) was added to dilute the mixture, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-83.

[0463] LC-MS (ESI) [M+H] + 355.1.

[0464] Reference Example 84: Preparation of Intermediate I-84

[0465]

[0466] Intermediate I-42 (880.00 mg, 1.83 mmol) was dissolved in concentrated hydrochloric acid (5 mL) at 25°C and stirred at 80°C for 16 hours. The mixture was concentrated to dryness, basified by the addition of ammonia in methanol (1 mL, 2 M), and concentrated under reduced pressure to afford the crude product. The crude product was purified by silica gel chromatography to afford Intermediate I-84.

[0467] LC-MS (ESI) [M+H] + 377.2.

[0468] Reference Example 85: Preparation of Intermediate I-85

[0469]

[0470] Intermediate I-85 (670.00 mg, 1.78 mmol) was dissolved in dichloromethane (10 mL) at 25°C. Boc-L-hydroxyproline (493.30 mg, 2.13 mmol), N,N-diisopropylethylamine (683.57 mg, 5.33 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (809.91 mg, 2.13 mmol) were added and stirred at room temperature for 1 hour. Water (20 mL) was added, and the mixture was extracted with dichloromethane (20 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-85.

[0471] LC-MS (ESI) [M+H] + 590.2.

[0472] Reference Example 86: Preparation of Intermediates I-86A and 86B

[0473]

[0474] Intermediate I-85 (1.22 g) was purified twice by SFC chiral separation columns (ChiralPak AD, 250×30 mm ID, 10 μm and SSwhelk O1, 250×30 mm ID, 10 μm) to obtain chiral intermediate I-86A (Rt=4.726 min) and chiral intermediate I-86B (Rt=5.077 min).

[0475] Chiral analysis method: Column: Chiralpak AD-3 150×4.6mm ID, 3μM

[0476] Mobile phase: A: supercritical carbon dioxide B: ethanol (0.05% diethylamine)

[0477] Elution gradient: 5% to 40% B, 5 minutes; 40% B 2.5 minutes; then 5% B 2.5 minutes

[0478] Flow rate: 2.5 mL / min

[0479] Column temperature: 35°C

[0480] Automatic Back Pressure Regulator (ABPR): 1500 psi

[0481] Intermediate I-86A LC-MS (ESI) [M+H] + 590.4; Intermediate I-86B LC-MS (ESI) [M+H] + 590.2.

[0482] Reference Example 87: Preparation of Intermediate I-87

[0483]

[0484] Intermediate I-86A (200.00 mg, 0.34 mmol) was dissolved in concentrated hydrochloric acid (3 mL) at 25°C and stirred at 100°C for 16 hours. The organic solvent was then removed by direct concentration to afford the crude chiral intermediate I-87. This crude product was used directly in the next reaction.

[0485] LC-MS (ESI) [M+H] + 223.0.

[0486] Reference Example 88: Preparation of Intermediate I-88

[0487]

[0488] Intermediate I-87 (400.00 mg) was dissolved in tetrahydrofuran (4 mL) at 25°C, and saturated aqueous sodium bicarbonate (2 mL) and di-tert-butyl dicarbonate (89.05 mg, 0.41 mmol) were added. The mixture was allowed to react at room temperature for 1 hour. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-88.

[0489] LC-MS (ESI) [M+H] + 323.0.

[0490] Reference Example 89: Preparation of Intermediate I-89

[0491]

[0492] At 25°C, intermediate I-88 (60.00 mg, 0.19 mmol) was dissolved in dichloromethane (5 mL). Triethylamine (56.42 mg, 0.56 mmol) and 2-methyl-4-nitrobenzoyl chloride (55.64 mg, 0.28 mmol) were added and stirred at room temperature for 16 hours. Water (10 mL) was added and the mixture was extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-89.

[0493] LC-MS (ESI) [M-100+H] + 386.0.

[0494] Reference Example 90: Preparation of Intermediate I-90

[0495]

[0496] At 25°C, intermediate I-89 (60.00 mg, 0.12 mmol) was dissolved in ethanol (8 mL). Reduced iron powder (34.47 mg, 0.62 mmol) and saturated aqueous ammonium chloride (2 mL) were added and stirred at 80°C for 5 hours. The mixture was filtered, and the filter cake was washed with ethyl acetate (20 mL). The filtrate was collected, water (10 mL) was added, and the layers were separated. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-90.

[0497] LC-MS (ESI) [M-100+H] + 356.4.

[0498] Reference Example 91: Preparation of Intermediate I-91

[0499]

[0500] At 25°C, intermediate I-90 (50.00 mg, 0.11 mmol) was dissolved in dichloromethane (5 mL). Triethylamine (33.29 mg, 0.33 mmol) and o-methylbenzoyl chloride (25.43 mg, 0.16 mmol) were added sequentially, and the mixture was stirred at room temperature for 1 hour. Water (10 mL) was added, and the mixture was extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-91.

[0501] LC-MS (ESI) [M-100+H] + 474.0.

[0502] Reference Example 92: Preparation of Intermediate I-92

[0503]

[0504] Intermediate I-86B (750.00 mg, 1.27 mmol) was dissolved in concentrated hydrochloric acid (8 mL) at 25°C and stirred at 100°C for 16 hours. The organic solvent was then concentrated to give the crude chiral intermediate I-92. The crude product was used directly in the next reaction.

[0505] LC-MS (ESI) [M+H] + 223.0.

[0506] Reference Example 93: Preparation of Intermediate I-93

[0507]

[0508] At 25°C, intermediate I-92 (1.40 g) was dissolved in tetrahydrofuran (20 mL). Saturated aqueous sodium bicarbonate (10 mL) and di-tert-butyl dicarbonate (332.61 mg, 1.52 mmol) were added and stirred at room temperature for 1 hour. Water (20 mL) was added and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-93.

[0509] LC-MS (ESI) [M+H] + 323.2.

[0510] Reference Example 94: Preparation of Intermediate I-94

[0511]

[0512] At 25°C, intermediate I-93 (70.00 mg, 0.22 mmol) was dissolved in dichloromethane (5 mL). Triethylamine (65.77 mg, 0.65 mmol) and 2-methyl-4-nitrobenzoyl chloride (64.92 mg, 0.33 mmol) were added and stirred at room temperature for 16 hours. Water (10 mL) was added and the mixture was extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-94.

[0513] LC-MS (ESI) [M-100+H] + 386.0.

[0514] Reference Example 95: Preparation of Intermediate I-95

[0515]

[0516] Intermediate I-94 (90.00 mg, 0.19 mmol) was dissolved in ethanol (8 mL) at 25°C. Reduced iron powder (51.71 mg, 0.93 mmol) and saturated aqueous ammonium chloride (2 mL) were added and stirred at 80°C for 5 hours. The mixture was filtered, and the filter cake was washed with ethyl acetate (20 mL). The filtrate was collected, water (20 mL) was added, and the layers were separated. The organic phase was collected and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain Intermediate I-95.

[0517] LC-MS (ESI) [M-100+H] + 356.2.

[0518] Reference Example 96: Preparation of Intermediate I-96

[0519]

[0520] At 25°C, intermediate I-95 (70.00 mg, 0.15 mmol) was dissolved in dichloromethane (5 mL). Triethylamine (46.60 mg, 0.46 mmol) and o-methylbenzoyl chloride (35.60 mg, 0.23 mmol) were added sequentially, and the mixture was stirred at room temperature for 1 hour. Water (10 mL) was added, and the mixture was extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-96.

[0521] LC-MS (ESI) [M-100+H] + 474.2.

[0522] Reference Example 97: Preparation of Intermediate I-97

[0523]

[0524] Under an ice-water bath, thionyl chloride (311 mg, 2.61 mmol) was added to a solution of intermediate I-19 (335 mg, 1.31 mmol) in N,N-dimethylacetamide (15 mL). The reaction mixture was stirred at room temperature for 3 hours, followed by the addition of intermediate I-5 (150 mg, 0.653 mmol). The reaction mixture was stirred and reacted at room temperature for 16 hours. Water (40 mL) was added for dilution, followed by extraction with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography to obtain intermediate I-97.

[0525] LCMS (ESI) [M+H] + 468.1.

[0526] Reference Example 98: Preparation of Intermediate I-98

[0527]

[0528] At room temperature, o-chlorobenzoyl chloride (1.27 g, 7.23 mmol) was added to a solution of methyl 6-aminonicotinate (1.00 g, 6.57 mmol) in pyridine (20.0 mL). The reaction mixture was stirred at room temperature for 1 hour, then poured into water (100 mL), filtered, and the filter cake dried to yield Intermediate I-98.

[0529] LC-MS (ESI) [M+H] + 290.9.

[0530] Reference Example 99: Preparation of Intermediate I-99

[0531]

[0532] Aqueous sodium hydroxide (2 mL, 3.5 N) was added to a solution of intermediate I-98 (680 mg, 2.34 mmol) in tetrahydrofuran (10 mL) at room temperature, and the reaction mixture was stirred at 70°C for 1 hour. The reaction mixture was adjusted to pH 5 with dilute hydrochloric acid (1 N), filtered, and the filter cake was drained to obtain intermediate I-99.

[0533] LC-MS (ESI) [M+H] + 277.0.

[0534] Reference Example 100: Preparation of Intermediate I-100

[0535]

[0536] Under an ice-water bath, thionyl chloride (622 mg, 5.23 mmol) was added to a solution of intermediate I-99 (800 mg, 2.89 mmol) in N,N-dimethylacetamide (20 mL). The reaction mixture was stirred at room temperature for 3 hours, followed by the addition of intermediate I-5 (300 mg, 1.31 mmol). The reaction mixture was stirred and reacted at room temperature for 16 hours. Water (40 mL) was added for dilution, followed by extraction with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel chromatography to obtain intermediate I-100.

[0537] LCMS (ESI) [M+H] + 487.9.

[0538] Preparation of Example:

[0539] Example 1: Preparation of Compound 1

[0540]

[0541] Intermediate I-8 (150 mg, 0.287 mmol) was added to an aqueous ammonia solution (8.00 mL) at room temperature. Elemental iodine (728 mg, 2.87 mmol) and α-isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylene) (1.00 mL) were added to the reaction system. The reaction mixture was stirred at room temperature for 24 hours. Saturated aqueous sodium thiosulfate (50 mL) was added and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC (ammonia system) to obtain compound 1.

[0542] LC-MS (ESI) [M+H] + 537.0.

[0543] 1 HNMR (400MHz, DMSO-d6) δ11.25–11.15(m,1H),8.55–8.13(m,1H),8.10–7.48(m,7H),7.27(d,J=7.4Hz ,1H),7.12–6.86(m,1H),5.09–4.71(m,1H),3.19–2.94(m,1H),2.88–2.54(m,2H),2.45–2.16(m,2H).

[0544] Example 2: Preparation of Compound 2

[0545]

[0546] Intermediate I-10 (150 mg, 0.312 mmol) was added to an aqueous ammonia solution (8.00 mL) at room temperature. Elemental iodine (792 mg, 3.12 mmol) and α-isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylene) (1.00 mL) were then added to the reaction system. The reaction mixture was stirred at room temperature for 24 hours. Saturated aqueous sodium thiosulfate (50 mL) was added and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC (ammonia system) to obtain compound 2.

[0547] LC-MS (ESI) [M+H] + 496.0.

[0548] 1 HNMR(400MHz,DMSO-d6)δ10.16-10.48(m,1H),7.55-7.80(m,2H),7.04-7.50(m,7H),6.59-6.82(m,1H),4.52 -4.98(m,1H),3.54-3.74(m,1H),3.34-3.49(m,1H),2.96-3.12(m,1H),2.72-2.84(m,1H),2.13-2.45(m,8H).

[0549] Example 3: Preparation of Compound 3

[0550]

[0551] At room temperature, intermediate I-17 (40 mg, 0.080 mmol) was added to aqueous ammonia (5 mL), followed by iodine (101.8 mg, 0.800 mmol) and α-isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylene) (0.1 mL). After the addition was complete, the reaction mixture was stirred at room temperature overnight. The mixture was diluted with saturated aqueous sodium sulfite (10 mL) and extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with water (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by preparative HPLC (ammonia system) to obtain compound 3.

[0552] LC-MS (ESI) [M+H] + 515.0.

[0553] 1H NMR(400MHz,DMSO-d6)δ11.28-10.84(m,1H),8.64-6.69(m,9H),5.00-4.67(m,1H),2.78 (dt,J=43.7,22.6Hz,1H),2.47(s,3H),2.22-1.74(m,5H),1.54(dd,J=52.0,31.2Hz,2H).

[0554] Example 4: Preparation of Compound 4

[0555]

[0556] Intermediate I-21 (100 mg, 0.232 mmol) was dissolved in aqueous ammonia (5.00 mL) at room temperature. Elemental iodine (177 mg, 0.696 mmol) and α-isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylene) (0.300 mL) were added to the reaction system. The reaction mixture was stirred at room temperature for 16 hours. Water (20 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated sodium thiosulfate (30 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC (ammonia system) to obtain compound 4.

[0557] LC-MS (ESI) [M+H] + 447.1.

[0558] 1 H NMR (400MHz, DMSO-d6) δ10.86–10.76(m,1H),8.48–8.01(m,2H),7.61–7.09(m,7H),6.85(dd,J=43.8,8. 3Hz,1H),4.86(d,J=12.6Hz,1H),2.84–2.71(m,1H),2.35(s,3H),2.13–1.77(m,5H),1.58–1.48(m,1H).

[0559] Example 5: Preparation of Compound 5

[0560]

[0561] At room temperature, intermediate I-23 (100 mg, 0.206 mmol) was added to aqueous ammonia (5 mL), followed by iodine (533 mg, 2.1 mmol) and α-isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylene) (0.3 mL). After the addition was complete, the reaction mixture was stirred at room temperature overnight. The mixture was diluted with saturated aqueous sodium sulfite (10 mL) and extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with water (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by preparative HPLC (ammonia system) to obtain compound 5.

[0562] LC-MS (ESI) [M+H] + 501.0.

[0563] 1 H NMR(400MHz,DMSO-d6)δ11.21–11.10(m,1H),8.50–7.93(m,2H),7.84–7.10(m,7H),6.96– 6.76(m,1H),4.96–4.71(m,1H),2.86–2.69(m,1H),2.21–1.83(m,5H),1.80–1.44(m,2H).

[0564] Example 6: Preparation of Compound 6

[0565]

[0566] Intermediate I-30 (150 mg, 0.336 mmol) was dissolved in aqueous ammonia (5.00 mL) at room temperature. Elemental iodine (426 mg, 1.68 mmol) and α-isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylene) (0.300 mL) were added to the reaction system. The reaction mixture was stirred at room temperature for 16 hours. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated sodium thiosulfate (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC (aqueous ammonia system) to obtain compound 6.

[0567] LC-MS (ESI) [M+H] + 461.1.

[0568] 1H NMR (400MHz, DMSO-d6) δ10.97–10.44(m,1H),8.59–7.03(m,8H),6.75(dd,J=41.4,8.3Hz,1H),4.97– 4.75(m,1H),2.90–2.63(m,1H),2.48–2.31(m,6H),2.22–1.74(m,5H),1.54(dd,J=53.9,30.7Hz,2H).

[0569] Example 7: Preparation of Compound 7

[0570]

[0571] Potassium tert-butoxide (85.3 mg, 0.760 mmol) and trimethylsulfoxide iodide (335 mg, 1.52 mmol) were added to dimethyl sulfoxide (1.5 mL) at room temperature and stirred at room temperature for 10 minutes. Intermediate I-31 (70 mg, 0.152 mmol) was then added, and the reaction mixture was reacted at 50°C for 16 hours. After cooling to room temperature, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined. The organic phases were washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC (ammonium bicarbonate system) to obtain compound 7.

[0572] LC-MS (ESI) [M+H] + 476.1.

[0573] 1 H NMR(400MHz,DMSO-d6)δ8.43–7.64(m,1H),7.53–6.91(m,7H),6.88–6.51(m,1H),4.94–4.43(m,1H),3 .39(d,J=9.5Hz,1H),3.26(d,J=12.6Hz,2H),3.18–2.77(m,2H),2.29–1.90(m,8H),1.84–1.46(m,2H).

[0574] Example 8: Preparation of Compound 8

[0575]

[0576] Intermediate I-34 (150 mg, 0.322 mmol) was dissolved in aqueous ammonia (6 mL) at room temperature. Elemental iodine (817 mg, 3.22 mmol) and α-isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylene) (0.2 mL) were added to the reaction system. The reaction mixture was stirred at room temperature for 16 hours. The reaction solution was quenched with saturated sodium sulfite (10 mL) and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC to obtain compound 8.

[0577] LC-MS (ESI) [M+H] + 479.9.

[0578] 1 H NMR (400MHz, DMSO-d6) δ10.66–10.31(m,1H),7.74–6.99(m,9H),6.78–6.56(m,1H),4. 84-3.63(m,1H),3.14–2.65(m,1H),2.39(s,3H),2.19–1.72(m,5H),1.69–1.45(m,2H).

[0579] Example 9: Preparation of Compound 9

[0580]

[0581] Compound I-36 (300 mg, 0.674 mmol) was dissolved in aqueous ammonia (20.0 mL) at room temperature. Elemental iodine (513 mg, 2.02 mmol) and α-isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylene) (30 mg) were added to the reaction system, and the reaction mixture was stirred at room temperature for 8 hours. The reaction system was poured into a saturated sodium bisulfite solution (100 mL) and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC to obtain compound 9.

[0582] LC-MS (ESI) [M+H] + 460.0.

[0583] 1H NMR(400MHz,DMSO-d6)δ10.49–10.10(m,1H),7.92–6.91(m,9H),6.79–6.53(m,1H),4.95– 3.61(m,1H),3.10–2.67(m,1H),2.40–2.33(m,6H),2.12–1.75(m,4H),1.69–1.43(m,2H).

[0584] Example 10: Preparation of Compounds 10A and 10B

[0585]

[0586] Intermediate I-37 (220 mg, 0.477 mmol) was dissolved in tert-butanol (2.20 mL) at room temperature. Trimethylsulfoxide iodide (1.05 g, 4.77 mmol) and potassium tert-butoxide (268 mg, 2.39 mmol) were added sequentially. The reaction mixture was stirred at 55°C for 18 hours. The reaction system was cooled to room temperature, poured into water (30.0 mL), and extracted with ethyl acetate (10 mL x 4). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC to obtain Compound 10A and Compound 10B.

[0587] Compound 10A:

[0588] LC-MS(ESI)[M+Na] + 497.0.

[0589] 1 H NMR (400MHz, DMSO-d6) δ10.41–10.28(m,1H),7.83–7.54(m,3H),7.48–7.22(m,7H),4.74–4.42( m,2H),3.54(m,1H),3.15–3.01(m,1H),2.96–2.80(m,1H),2.41–2.31(m,8H),1.87–1.52(m,3H).

[0590] Compound 10B:

[0591] LC-MS (ESI) [M+H] + 489.1.

[0592] 1H NMR(400MHz, DMSO-d6+D2O)δ7.70–7.56(m,1H),7.41(dd,J=8.3,2.4Hz,1H),7.33–6.87(m,8H),4.62–4.33(m,2H), 3.41–3.30(m,2H),3.28–3.20(m,1H),3.09–2.96(m,1H),2.88–2.72(m,1H),2.30–2.06(m,8H),1.82–1.36(m,3H).

[0593] Example 11: Preparation of Compound 11

[0594]

[0595] Intermediate I-47 (40.00 mg, 0.070 mmol) was dissolved in a solution of hydrogen chloride in dioxane (5 mL, 3 M) at 25°C and reacted at room temperature for 1 hour. The mixture was concentrated to dryness and purified by preparative HPLC (ammonium bicarbonate system) to obtain compound 11.

[0596] LC-MS (ESI) [M+H] + 474.2.

[0597] 1 H NMR (400MHz, Methanol-d4) δ7.68–7.51(m,1H),7.48–7.08(m,5H),6.92(dd,J=8.5,2.5Hz ,1H),6.73(d,J=8.5Hz,1H),6.60(d,J=8.4Hz,1H),6.09(dt,J=64.2,6.5Hz,1H),4.65–4.5 5(m,1H),3.71(s,1H),3.46(dt,J=13.0,5.2Hz,1H),2.89–2.78(m,2H),2.78–2.64(m,1H) ,2.62–2.51(m,1H),2.51–2.40(m,1H),2.30–2.28(m,6H),2.24–2.06(m,1H),1.97(m,1H).

[0598] Example 12: Preparation of Compound 12

[0599]

[0600] 2-Methylbenzoyl chloride (21.9 mg, 0.142 mmol) and triethylamine (28.7 mg, 0.284 mmol) were added to a solution of intermediate I-57 (35 mg, 0.0944 mmol) in dichloromethane (5 mL) at room temperature. The reaction mixture was allowed to react at room temperature for 3 hours. The reaction mixture was diluted with dichloromethane (10 mL), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by preparative HPLC (ammonium bicarbonate system) to obtain compound 12.

[0601] LC-MS (ESI) [M+H] + 489.1

[0602] 1 H NMR (400MHz, DMSO-d6) δ10.41–10.26(m,1H),7.77–6.58(m,10H),4.37–4.04(m,1H),3.99–3.47(m, 4H),3.06–2.80(m,1H),2.70–2.51(m,1H),2.45–2.16(m,7H),2.04–1.93(m,1H),1.86–1.51(m,3H).

[0603] Example 13: Preparation of Compound 13

[0604]

[0605] At room temperature, intermediate I-7 (54.0 mg, 0.174 mmol) was dissolved in N,N-dimethylacetamide (5.00 mL). The temperature was lowered to 0°C, and thionyl chloride (20.7 mg, 0.174 mmol) was added under argon. The reaction mixture was stirred at 0°C for 3 hours, and then intermediate I-58 (30.0 mg, 0.116 mmol) was added. The reaction mixture was stirred at room temperature for another 16 hours. The mixture was added to water (20 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC (ammonia system) to obtain compound 13.

[0606] LCMS (ESI) [M+H] + 552.0.

[0607] 1HNMR(400MHz,DMSO-d6)δ11.29–11.19(m,1H),8.41–7.97(m,2H),7.90–7.63(m, 6H),7.45–6.92(m,2H),4.85–4.49(m,3H),3.54–2.63(m,4H),2.38–2.15(m,1H).

[0608] Example 14: Preparation of Compound 14

[0609]

[0610] Potassium tert-butoxide (175 mg, 1.56 mmol) and trimethylsulfoxide iodide (343 mg, 1.56 mmol) were added to dimethyl sulfoxide (1.00 mL) at room temperature and stirred at room temperature for 5 minutes. Intermediate I-59 (100 mg, 0.223 mmol) was then added, and the reaction mixture was stirred at 70°C for 20 hours. After cooling to room temperature, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (5 mL x 3). The organic phases were combined. The organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by preparative HPLC (ammonia system) to obtain compound 14.

[0611] LC-MS (ESI) [M+H] + 462.0.

[0612] 1 H NMR (400MHz, DMSO-d6) δ11.09–10.74(m,1H),8.56–8.00(m,2H),7.87–7.17(m,7H),7.06–6. 78(m,1H),4.80–4.33(m,3H),3.11–2.92(m,1H),2.89–2.54(m,2H),2.47–2.15(m,4H),2.12– 1.55(m,3H).

[0613] Example 15: Preparation of Compound 15

[0614]

[0615] At room temperature, intermediate I-65 (200 mg, 0.392 mmol) was dissolved in dimethyl sulfoxide (5.00 mL). Trimethylsulfoxide iodide (863 mg, 3.92 mmol) and potassium tert-butoxide (220 mg, 1.96 mmol) were added sequentially. The reaction mixture was stirred at 50°C for 24 hours. The reaction system was cooled to room temperature, poured into water (30 mL), and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over saturated brine (15 mL), and anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by preparative HPLC (ammonium bicarbonate system) to obtain compound 15.

[0616] LC-MS (ESI) [M+H] + 524.0.

[0617] 1 H NMR(400MHz,DMSO-d6)δ8.12–7.65(m,1H),7.64–7.10(m,10H),7.08–6.01(m,4 H),4.88–4.43(m,1H),3.14–2.86(m,4H),2.83–2.66(m,1H),2.17–1.51(m,4H).

[0618] Example 16: Preparation of Compound 16

[0619]

[0620] O-(7-Azabenzotriazol-1-yl)-N,N,N'-tetramethyluronium hexafluorophosphate (46.8 mg, 0.123 mmol) and N,N-diisopropylethylamine (23.9 mg, 0.185 mmol) were added to a solution of intermediate I-71 (21 mg, 0.0616 mmol) and o-methylbenzoic acid (10.1 mg, 0.0742 mmol) in N,N-dimethylformamide (5 mL) at room temperature. The reaction mixture was stirred at 50°C for 16 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness and purified by C18 reverse-phase chromatography to yield compound 16.

[0621] LC-MS (ESI) [M+H] + 459.2.

[0622] 1H NMR (400MHz, DMSO-d6) δ10.40–10.24(m,1H),7.77–7.45(m,2H),7.40–7.02(m,7H),6.76–6.66(m,1H),4.88–2.68( m,1H),2.37(m,6H),2.00–1.87(m,1H),1.82–1.52(m,2H),1.38–1.14(m,2H),1.02–0.85(m,1H),0.82–0.68(m,2H).

[0623] Example 17: Preparation of Compound 17

[0624]

[0625] 2-Methylbenzoic acid (19.3 mg, 0.142 mmol), N,N-diisopropylethylamine (43.7 mg, 0.339 mmol), and O-(7-azabenzotriazol-1-yl)-N,N,N'-tetramethyluronium hexafluorophosphate (85.9 mg, 0.226 mmol) were added to a solution of intermediate I-83 (40 mg, 0.113 mmol) in N,N-dimethylformamide (3 mL) at room temperature. The reaction mixture was reacted at room temperature for 16 hours. The reaction mixture was separated and purified by preparative HPLC (formic acid system) to obtain compound 17.

[0626] LC-MS (ESI) [M+H] + 473.3.

[0627] 1 H NMR (400MHz, DMSO-d6) δ10.40–10.26(m,1H),7.75–7.17(m,9H),6.86–6.55(m,1H),4.52– 3.50(m,1H),2.98–2.83(m,1H),2.77–2.64(m,1H),2.41–2.23(m,7H),2.22–1.35(m,8H).

[0628] Example 18: Preparation of Compound 18

[0629]

[0630] Intermediate I-91 (50.00 mg, 0.087 mmol) was dissolved in a solution of hydrogen chloride in dioxane (5 mL, 3 M) at 25°C and stirred at room temperature for 1 hour. The mixture was concentrated to dryness and purified by preparative HPLC (ammonium bicarbonate system) to give chiral compound 18.

[0631] LC-MS (ESI) [M+H]+ 474.2.

[0632] 1 H NMR(400MHz,DMSO-d6)δ10.31(s,1H),7.58–7.21(m,6H),7.08(dd,J=8.3,2.5Hz ,1H),6.83(d,J=8.3Hz,1H),6.69(d,J=8.4Hz,1H),6.32–5.90(m,1H),4.68–4.4 9(m,1H),3.77–3.64(m,1H),3.59(s,1H),3.49–3.40(m,2H),2.82–2.69(m,2H), 2.44(d,J=6.2Hz,2H),2.39(s,1H),2.34(s,3H),2.29(s,3H),2.22–2.06(m,1H).

[0633] Example 19: Preparation of Compound 19

[0634]

[0635] Intermediate I-96 (70.00 mg, 0.12 mmol) was dissolved in a solution of hydrogen chloride in dioxane (5 mL, 3 M) at 25°C and stirred at room temperature for 1 hour. The mixture was concentrated to dryness and purified by preparative HPLC (ammonium bicarbonate system) to give chiral compound 19.

[0636] LC-MS (ESI) [M+H] + 474.2.

[0637] 1 H NMR(400MHz,DMSO-d6)δ10.34(s,1H),7.57–7.23(m,6H),7.11(dd,J=8.5,2.4Hz,1 H),6.85(d,J=8.5Hz,1H),6.66(d,J=8.4Hz,1H),6.27–5.98(m,1H),4.63–4.48(m, 1H),3.85–3.57(m,1H),3.51–3.34(m,1H),2.99–2.78(m,3H),2.64(q,J=8.6,7.7H z,1H),2.48(d,J=18.7Hz,2H),2.39(s,1H),2.34–2.28(m,6H),2.23–1.90(m,1H).

[0638] Example 20: Preparation of Compound 20

[0639]

[0640] α-Isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylenediamine) (1.00 mL) and elemental iodine (1.19 g, 4.69 mmol) were added to a solution of intermediate I-97 (220 mg, 0.470 mmol) in aqueous ammonia (20 mL) at room temperature. The reaction mixture was stirred at room temperature for 16 hours (LCMS and TLC indicated approximately 10% product formation, with most of the starting material remaining). Saturated aqueous sodium sulfite (40 mL) was added, and the mixture was extracted with ethyl acetate (30 mL x 2). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel chromatography to recover a mixture of starting material and product (210 mg). The above procedure was repeated twice. The reaction mixture was added to a saturated aqueous sodium sulfite solution (30 mL), extracted with ethyl acetate (30 mL x 2), and the combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel chromatography and C18 reverse phase chromatography to obtain compound 20.

[0641] LC-MS (ESI) [M+H] + 483.1.

[0642] 1 H NMR(400MHz,DMSO-d6)δ10.90–10.80(m,1H),8.48–8.07(m,1H),8.08–7.20(m,8H),7.09– 6.82(m,1H),5.03–4.76(m,1H),3.19–2.92(m,1H),2.84–2.54(m,2H),2.42–2.13(m,5H).

[0643] Example 21: Preparation of Compound 21

[0644]

[0645] α-Isotridecyl-ω-hydroxy-poly(oxy-1,2-ethylenediamine) (2.00 mL) and elemental iodine (1.56 g, 6.15 mmol) were added to a solution of I-100 (300 mg, 0.614 mmol) in aqueous ammonia (20 mL) at room temperature. The reaction mixture was stirred at room temperature for 16 hours (LCMS and TLC showed approximately 10% product formation, with most of the starting material remaining). Saturated aqueous sodium sulfite (40 mL) was added, and the mixture was extracted with ethyl acetate (30 mL x 2). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel chromatography to recover a mixture of starting material and product (295 mg). The reaction mixture was then re-dosed as described above. Saturated aqueous sodium sulfite (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL x 2). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel chromatography and C18 reverse phase chromatography to obtain compound 21.

[0646] LC-MS (ESI) [M+H] + 503.0.

[0647] 1 H NMR (400MHz, DMSO-d6) δ11.16–11.06(m,1H),8.56–6.84(m,10H),5.08–4.57(m,1H),3.18–2.94(m,1H),2.89–2.53(m,2H),2.46–2.16(m,2H).

[0648] Experimental Example 1: Inhibitory IC of Compounds on Vasopressin-Induced Activation of Vasopressin Receptor V2R 50 test

[0649] (1) Cells

[0650] HeLa cell line stably expressing human vasopressin receptor V2R (HeLa-V2R): constructed by Shanghai GeneChem Technology Co., Ltd. using lentiviral infection method, and verified by qPCR to stably express human V2R.

[0651] (2) Reagents

[0652] DMEM cell culture medium: Brand: Gibco, Catalog Number: 11995065; Fetal bovine serum: Brand: Jitai, Catalog Number: FND500; 0.25% trypsin: Brand: Gibco, Catalog Number: 25200072; Puromycin Dihydrochloride: Brand: Gibco, Catalog Number: A1113803; cAMP-GS HIRANGE KIT: Brand: Cisbio, Catalog Number: 62AM6PEC; IBMX: Brand: Sigma, Catalog Number: i5879; Vasopressin AVP: Customized by Jier Biochemical (Shanghai) Co., Ltd.

[0653] (3) Test method

[0654] HeLa-V2R cells were cultured in DMEM supplemented with 10% fetal bovine serum at 37°C and 5% CO2. 2 μg / mL puromycin was added to the culture medium to continuously select cells expressing V2R. On the day of the experiment, the cells were trypsinized, washed twice with the stimulation buffer in the cAMP-GS HIRANGE kit, resuspended, counted, and prepared into 1.6×10 6 cells / ml, add IBMX to a final concentration of 0.5mM. Transfer 5uL of cell suspension / well to a 384-well plate, and add 2.5uL of different concentrations of the test compound (3-fold dilution starting from 10uM, 10 concentration gradients) or DMSO (minimum value Min, maximum value Max control) to the corresponding wells. After incubation at room temperature for 30 minutes, add 2.5uL of vasopressin AVP solution to the test compound well and the maximum value well to a final concentration of 2.25nM, add 2.5uL of stimulation buffer to the minimum value well, and incubate at 25 degrees for 60 minutes. At the same time, prepare cAMP standard samples (3-fold dilution starting from 5.6uM, 10 concentration points), and transfer 10uL of cAMP standard to the corresponding wells of the 384-well plate. Dilute the cAMP-d2 fluorescent and anti-cAMP antibody probes provided in the cAMP-GS HIRANGE kit 20-fold with the lysis buffer in the kit. Add 5 μL of each to each well of a 384-well plate. Mix thoroughly, centrifuge briefly, and incubate at 25°C for 2 hours before analysis. Analyze samples using the HTRF method on an Envision microplate reader, measuring fluorescence intensity at 615 nm and 665 nm. Duplicate wells are prepared for each sample, and 32 replicate wells are prepared for both the Min and Max wells.

[0655] (4) Data processing

[0656] Calculate the fluorescence intensity ratio FI of each well sample at 665nm and 615nm wavelength 665 / 615 The logarithm of the standard concentration is X, FI 665 / 615X1000 is the Y value, and the standard curve is obtained by fitting the "log (inhibitor) vs response–variable slope (four parameters)" model in Prism 8.0 software. 665 / 615 X1000 is the Y value, and the cAMP concentration corresponding to each sample was calculated based on the above standard curve in Prism8.0 software.

[0657] The formula for calculating %Inhibition is as follows:

[0658]

[0659] in is the average calculated value of cAMP concentration in all maximum value wells; is the average calculated value of cAMP concentration in all minimum value wells; Ccmpd is the calculated value of cAMP concentration of the test compound.

[0660] IC50 was calculated using nonlinear regression using the "log (inhibitor) vs response – variable slope (four parameters)" model in Prism 8.0 software, with % Inhibition as Y and the logarithm of compound concentration as X, where Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * Hill Slope)).

[0661] The experimental results are shown in Table 1:

[0662] Table 1: Evaluation of the compounds' inhibition of cAMP increase in human cervical cancer cells (Human V2R Hela-Stable cell line OE2)

[0663] Compound number <![CDATA[IC 50 (nM)]]> Compound number <![CDATA[IC 50 (nM)]]> 1 10.88 2 9.35 4 3.14 5 4.67 8 5.01 9 4.88 16 11.23 17 10.43 20 5.54 21 4.51

[0664] Experimental Example 2: In vivo pharmacokinetic study of the compounds of the present invention

[0665] In this study, the pharmacokinetics of the drug were evaluated in mice via intravenous and oral administration.

[0666] Experimental Methods and Conditions: Male CD1 mice, 6-8 weeks old, with free access to food and water, were given a single intravenous injection of the test compound (1 mg / kg) in 5% DMSO / 10% Solutol / 85% Saline. Blood samples were collected retro-orbitally 5, 15, 30, 1, 2, 4, 8, or 24 hours after administration, or 10 mg / kg in 5% DMSO / 10% Solutol / 85% Saline (6, 8, or 24 hours after administration). Blood samples (≥50 μL each) were collected and anticoagulated with sodium heparin. Plasma was centrifuged within 1 hour for analysis. Plasma concentrations were determined by liquid chromatography-tandem mass spectrometry (LC / MS / MS), and pharmacokinetic parameters were calculated using Phoenix WinNonlin software. Tofaptane was used as the control substance 1. The experimental results are shown in Tables 2 and 3.

[0667] Table 2: Pharmacokinetics of oral administration (10 mg / kg)

[0668] Compound <![CDATA[T 1 / 2 (hr)]]> <![CDATA[C max (ng / mL)]]> <![CDATA[AUC 0‐inf (ng*hr / mL)]]> F(％) Compound 1 0.60 1255 4058 56 Reference substance 1 1.58 1307 1613 44

[0669] Table 3: Pharmacokinetics of intravenous administration (1 mg / kg)

[0670] Compound <![CDATA[T 1 / 2 (hr)]]> <![CDATA[AUC 0-inf (ng*hr / mL)]]> Cl (ml / min / kg) Compound 1 0.92 719 23.2

[0671] Reference substance 1 0.53 367 45.5

[0672] The experimental data showed that the pharmacokinetic results of the compound of the present invention in mice after intravenous and oral administration showed a lower metabolic clearance rate Cl and a higher in vivo exposure amount AUC 0-inf .

[0673] Experimental Example 3: Test of the inhibitory effect of compounds on LLC-PK1 cell proliferation

[0674] (1) Cells

[0675] Pig kidney epithelial cells LLC-PK1: purchased from ATCC, Cat#CL-101

[0676] (2) Reagents:

[0677] Medium 199,Gibco(Cat#11150059)

[0678] Fetal Bovine Serum(FBS),Australia,Jitai(Cat#FND500)

[0679] Trypsin-EDTA(0.25%),phenol red,Gibco(Cat#25200072)

[0680] PBS, pH 7.4, Gibco (Cat#10010031)

[0681] DMSO (dimethyl sulfoxide), Sigma (Cat#D8418)

[0682] Poly-D-lysine,Gibco(Cat#A3890401)

[0683] Vasopressin AVP: Customized by Jier Biochemical (Shanghai) Co., Ltd.

[0684] Verapamil hydrochloride,MCE(Cat#HY-A0064)

[0685] AlamarBlue TM HS Cell Viability Reagent, Invitrogen(Cat#A50100)

[0686] (3) Test method:

[0687] The pathogenesis of polycystic kidney disease (PKD) is associated with low intracellular calcium concentrations in renal collecting duct epithelial cells, leading to cAMP-dependent excessive cell proliferation. Based on a 2004 study by Tamio Yamaguchi et al. published in The Journal of Biological Chemistry, we optimized and performed a renal epithelial LLC-PK1 cell proliferation assay to evaluate the ability of compounds to inhibit vasopressin-induced cell proliferation after reducing intracellular calcium concentrations.

[0688] LLC-PK1 cells were cultured in M199 medium supplemented with 10% fetal bovine serum at 37°C and 5% CO2. On the first day of the experiment, a 96-well plate was coated with 0.01% Poly-D-lysine. 100 μl of culture medium was added to each well. After standing at room temperature for 10 minutes, the cells were aspirated and air-dried at room temperature for 1 hour. The cells were then washed once with 200 μl of 1X PBS and used for further filtration. LLC-PK1 cells were trypsinized, centrifuged, resuspended in serum-free M199, counted, and diluted to 1×10 cells in serum-free M199 medium. 5 / ml cell suspension, add FBS to a final concentration of 1%. Transfer 200ul of cell suspension / well to a 96-well plate. After 24 hours of cell culture, aspirate the supernatant of the solution, wash once with 200ul PBS, and then add 160ul of M199 culture medium containing 0.05% FBS and 20ul of 10X Verapamil (final concentration 5uM) in sequence, and continue to culture for 24 hours. On the third day, add 10uL of different concentrations of the test compound (final concentration starting from 3uM, 3-fold dilution, 8 concentration gradients) or DMSO (minimum value Min, maximum value Max control) to the corresponding wells. Add 10uL of vasopressin AVP solution to the test compound well and the maximum value well to a final concentration of 10nM, and add 10uL of serum-free M199 culture medium to the minimum value well, and continue to culture for 48 hours. On the fifth day, carefully aspirate the culture medium, wash the cells once with 200 μl of PBS, carefully add 90 μl of M199 serum-free culture medium, then add 10 μl of AlamarBlue reagent. Centrifuge at 300 rpm for 1 minute, incubate at 37°C for 2 hours, and then analyze. Samples were analyzed using a SpectraMax instrument with excitation at 560 nm and emission at 595 nm. Three replicate wells were performed for each sample, and six replicate wells were performed for both the Min and Max instruments.

[0689] (4) Data processing

[0690] Compound concentration was used as the X value, and the fluorescence intensity of each experimental well, after subtracting the background fluorescence intensity, was used as the Y value, representing the number of viable cells in that well at the time of the assay. A bar graph was constructed using GraphPad Prism 8.0 software, using a Grouped-Summary-Data-Separated bar graph, to demonstrate the dose-effect relationship between different compounds on AVP-induced cell proliferation. Using tolvaptan as a positive control, the inhibitory effect of the compounds on proliferation was qualitatively evaluated: compounds with overall superiority to tolvaptan were designated "+++," those with similar performance to tolvaptan were designated "++," those with weaker performance than tolvaptan were designated "+," and those with no inhibition were designated "-."

[0691] Data quality control: Calculate S / B, which is the average value of the Max well / the average value of the Min well. A value ≥ 2 is considered to have passed QC.

[0692] The experimental results showed that compound 1 had a stronger inhibitory effect on LLC-PK1 cell proliferation than the control substance 1 (tofaptane), and had no pro-proliferation effect under high concentration conditions. Figure 1 shown.

Claims

1. A compound represented by formula (II), an optical isomer thereof, and a pharmaceutically acceptable salt thereof, in, X2 is selected from CH and N; T1 and T2 are independently selected from N and CH; R1 is independently selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups; R 2a and R 2b are independently selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl and C 3-6 Cycloalkyl, the C 1-6 Alkyl or C 3-6 Cycloalkyl is optionally substituted with 1, 2 or 3 R; R3 is selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups; R4 are independently selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 3-6 Cycloalkyl, phenyl and 5-6 membered heteroaryl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 3-6 Cycloalkyl, phenyl or 5- to 6-membered heteroaryl is optionally substituted by 1, 2 or 3 R; R is selected from H, F, Cl, Br, I, CN, OH, NH2, C 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Alkylamino, the C 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 The alkylamino group is optionally substituted with 1, 2 or 3 R'; R' is selected from H, F, Cl, Br, I, CN, OH, NH2 and C 1-6 alkyl; Structural unit Selected from 2. The compound according to claim 1, its optical isomers and pharmaceutically acceptable salts thereof, wherein: The structure of the compound is shown in formula (III), 3. The compound according to claim 1 or 2, its optical isomers and pharmaceutically acceptable salts thereof, wherein: R is selected from H, F, Cl, Br, I, CN, OH, NH2, CH3, CF3, 4. The compound according to claim 1 or 2, its optical isomers and pharmaceutically acceptable salts thereof, wherein: R4 is selected from H, F, Cl, Br, I, CN, OH, NH2, CH3, CF3, cyclopropyl, cyclobutyl, cyclopentyl, phenyl, pyridyl, pyrimidinyl, thienyl and thiazolyl.

5. The compound according to claim 1, its optical isomers and pharmaceutically acceptable salts thereof, wherein: Structural unit Selected from 6. The compound of the following formula, its optical isomers and pharmaceutically acceptable salts thereof, which are selected from 7. Use of the compound according to any one of claims 1 to 6, its optical isomers and pharmaceutically acceptable salts thereof in the preparation of a medicament for preventing or treating diseases associated with the arginine vasopressin V1a receptor, arginine vasopressin V1b receptor, arginine vasopressin V2 receptor, sympathetic nervous system or renin-angiotensin-aldosterone system.

8. The use according to claim 7, wherein the diseases related to the arginine vasopressin V1a receptor, the arginine vasopressin V1b receptor, the arginine vasopressin V2 receptor, the sympathetic nervous system or the renin-angiotensin-aldosterone system include: High blood pressure, Reye's syndrome, dysmenorrhea, premature birth, impaired corticotropin-releasing hormone secretion, adrenal hyperplasia, depression, chronic congestive heart failure, cirrhosis, syndrome of inappropriate antidiuretic hormone secretion, hyponatremia due to chronic heart failure / cirrhosis / inappropriate antidiuretic hormone secretion, or polycystic kidney disease.

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