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WO2024169773A1 - 吲唑啉酮类化合物及其制备方法和用途 - Google Patents

吲唑啉酮类化合物及其制备方法和用途 Download PDF

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Publication number
WO2024169773A1
WO2024169773A1 PCT/CN2024/076228 CN2024076228W WO2024169773A1 WO 2024169773 A1 WO2024169773 A1 WO 2024169773A1 CN 2024076228 W CN2024076228 W CN 2024076228W WO 2024169773 A1 WO2024169773 A1 WO 2024169773A1
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mmol
reaction
alkyl
add
compound
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PCT/CN2024/076228
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English (en)
French (fr)
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柯潇
方群
孙启正
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成都康弘药业集团股份有限公司
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    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/10—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing aromatic rings
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
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    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
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    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
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    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
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    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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    • A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
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    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
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    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
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    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/10—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing aromatic rings
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
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    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
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    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
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    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
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    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
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    • C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
    • C07D493/08—Bridged systems

Definitions

  • This patent application relates to the field of medicine, and relates to indazolinone GLP1 receptor agonists, their preparation methods and their applications in medicine.
  • GLP1R Glucagon-like peptide 1 Receptor
  • GLP1R Glucagon-like peptide 1 Receptor
  • GLP1R agonists In the field of type 2 diabetes and obesity, many peptide GLP1R agonists have been launched. In the field of NASH and neurodegenerative diseases, GLP1R agonists have also shown preclinical/clinical therapeutic potential, and related clinical explorations are also being actively carried out. However, most of the peptide drugs currently on the market require subcutaneous injection, which affects patient compliance.
  • PDE10A is a dihydrolase that can hydrolyze cyclic adenosine monophosphate and cyclic guanosine monophosphate at the same time. It is highly expressed in the brain, especially in the medium spiny neurons of the striatum. Based on reports in the literature (Expert Rev Neurother, 2017 (17), 6, 553-560; Brain 2015, 138 (pt 10), 3003-3015), in clinical practice, PDE10A selective inhibitors represented by Mardepodect unexpectedly showed side effects of movement disorders. This result is consistent with the clinical observation that PDE10A expression in the striatum of Parkinson's and Huntington's patients is reduced. This suggests that the inhibition of PDE10A may be accompanied by motor side effects.
  • GLP-1 receptor agonists include WO2009111700A2, WO2010114824A1, WO2018109607A1, WO2019239319A1, and WO2018056453A1, etc. Therefore, it is necessary to provide a GLP1R agonist that can be taken orally, has good efficacy, and has few side effects.
  • the present invention provides a GLP1R agonist which can be taken orally, has good efficacy, few side effects, good solubility and high bioavailability.
  • R1 is selected from hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxyl, cycloalkyl, heterocyclic radical, aryl and heteroaryl, wherein said alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclic radical, aryl and heteroaryl are optionally substituted with one or more substituents selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxyl, hydroxyalkyl, cycloalkyl, heterocyclic radical, aryl and heteroaryl;
  • R2 is selected from hydrogen atom, alkyl, aryl or heteroaryl, wherein the alkyl, aryl or heteroaryl is optionally substituted by one or more substituents selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxyl, hydroxyalkyl, cycloalkyl, heterocyclic, aryl and heteroaryl, and the cycloalkyl and heterocyclic are optionally substituted by one or more substituents selected from halogen, alkyl, cyano;
  • R3 is the same or different at each occurrence and is independently selected from hydrogen, halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl and heteroaryl;
  • R5 is selected from cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally substituted with one or more substituents selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, hydroxyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;
  • X is selected from N, O or S;
  • L is a straight chain or branched chain alkyl
  • Z0, Z1, Z2, Z3, Z4, Z5 and Z6 may be the same or different, and each independently selects CR4 or a nitrogen atom;
  • n, and p are integers of 0-4.
  • R1 and R3 are each independently selected from a hydrogen atom, a halogen, a C1-6 alkyl, a C1-6 alkoxy, a halogenated C1-6 alkyl, a halogenated C1-6 alkoxy, a cyano group, an amino group, a nitro group, a hydroxyl group, a hydroxy C1-6 alkyl group, a C3-8 cycloalkyl group and a C3-20 heterocyclic group.
  • R2 is preferably a C1-6 alkyl substituted by an aryl or heteroaryl group, and the aryl or heteroaryl group is further preferably an aryl or heteroaryl group in the form of a monocyclic or condensed ring, optionally substituted by one or more substituents selected from halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, and hydroxyl.
  • R5 is preferably an aryl group or a heteroaryl group, and the aryl group or the heteroaryl group is optionally substituted by an alkyl group, an alkoxy group, a haloalkyl group, a cyano group, or a halogen group.
  • Another aspect of the present invention provides a compound having a structure as shown in Formula II or a pharmaceutically acceptable salt thereof:
  • Z0 is selected from C or N;
  • Z2 is selected from CR4 or N, R4 is selected from H or CH3;
  • Z4, Z5 or Z6 is selected from C or N;
  • R2 is selected from a heterocyclic group or a C1-6 alkyl group, wherein the heterocyclic group is preferably an optionally substituted C3-8 oxygen-containing heterocyclic group, and the C1-6 alkyl group is preferably substituted by CH2OCH3, Substituted C1-6 alkyl;
  • R3 is the same or different at each occurrence and is independently selected from hydrogen, F or methyl, and n is selected from an integer of 1 or 2;
  • L is CH2, and p is an integer of 1-4.
  • the aryl or heteroaryl group is selected from:
  • X0, X1, X2, X3 and X4 are each independently selected from C, N, O and S;
  • X0 is selected from C or N;
  • At least one of X1, X2, X3 and X4 is N;
  • aryl or heteroaryl groups are optionally substituted by H, halogen, cyano, amino, nitro, hydroxy, C1-6 alkyl, C1-6 alkoxy, C3-8 cycloalkyl, halogenated C1-6 alkyl, halogenated C1-6 alkoxy, or hydroxy C1-6 alkyl.
  • Another aspect of the present invention provides a compound having a structure as described in Formula III or a pharmaceutically acceptable salt thereof:
  • Z0 is selected from C or N;
  • Z2 is selected from CR4 or N, R4 is selected from H or CH3;
  • Z4, Z5 or Z6 is selected from C or N;
  • R3 is selected from F, n is selected from an integer of 1 or 2;
  • R5 is selected from the following aryl or heteroaryl groups:
  • X0 is selected from C or N
  • the aryl or heteroaryl group may be optionally substituted by F, Cl, CN, OCH3, CF3;
  • R6 is selected from -CH2OCH3, R
  • R10 is selected from H, halogen, CN, OCH3, or CH2CF2.
  • Another aspect of the present invention provides a compound having a structure shown in Formula IV or a pharmaceutically acceptable salt thereof:
  • Z0 is selected from C or N;
  • Z4, Z5 or Z6 is selected from N or C;
  • R5 is selected from the following groups:
  • X0 is selected from C or N
  • R7 is selected from F or methoxy
  • R8 is selected from Cl or CN
  • R12 is selected from halogen or CF3
  • R6 is selected from
  • Another aspect of the present invention provides a compound having a structure as described in Formula V or a pharmaceutically acceptable salt thereof:
  • Z6 is selected from C or N;
  • R5 is selected from the following groups:
  • X0 is selected from C or N
  • R7 is selected from F or methoxy
  • R8 is selected from Cl or CN
  • R12 is selected from halogen or CF3
  • R6 is selected from Best
  • Another aspect of the present invention provides a compound having a structure as described in Formula VI or a pharmaceutically acceptable salt thereof:
  • R13 is selected from hydrogen, halogen, C 1-3 alkyl, C 1-3 haloalkyl or C 1-3 alkoxy;
  • Z4, Z5 or Z6 is selected from N or C;
  • X0 is selected from C or N;
  • R7, R8, R14, R15 are selected from hydrogen, halogen, cyano, C1-3 alkyl, C1-3 haloalkyl or C1-3 alkoxy.
  • Another aspect of the present invention provides a compound having a structure as described in Formula VII or a pharmaceutically acceptable salt thereof:
  • R13 is selected from hydrogen, halogen, C 1-3 alkyl, C 1-3 haloalkyl or C 1-3 alkoxy;
  • Z6 is selected from N or C
  • R7 and R8 are selected from hydrogen, halogen, cyano, C 1-3 alkyl, C 1-3 haloalkyl or C 1-3 alkoxy.
  • R7 is selected from halogen or methoxy
  • R8 is selected from halogen or CN.
  • R7 is selected from F or methoxy
  • R8 is selected from Cl or CN.
  • compositions which contains the general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (VI), general formula (VII) and specific compounds in the present patent application, or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.
  • the present patent application further relates to the use of general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (VI), general formula (VII) and specific compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing the same in the preparation of drugs for stimulating GLP-1 receptors.
  • This patent application further relates to general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (VI), Use of general formula (VII) and specific compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same in the preparation of drugs for treating and/or preventing type I diabetes, type II diabetes, malnutrition-related diabetes, diabetic complications, obesity, hyperglycemia, glucose intolerance, cardiovascular disease, hyperlipidemia, cerebral infarction, stroke, non-alcoholic steatohepatitis (NASH), Parkinson's disease, dementia, insulin resistance and hepatic insulin resistance; preferably use of the same in the preparation of drugs for treating and/or preventing type I diabetes, type II diabetes, obesity, diabetic complications, non-alcoholic steatohepatitis and cardiovascular disease.
  • NASH non-alcoholic steatohepatitis
  • Parkinson's disease dementia
  • insulin resistance and hepatic insulin resistance preferably use of the same in the preparation of drugs for treating and/or preventing
  • the present patent application further relates to the use of general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (VI), general formula (VII) and specific compounds, or pharmaceutically acceptable salts thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same in the preparation of drugs for the treatment and/or prevention of idiopathic type I diabetes, latent immune diabetes in adults (LADA), maturity-onset diabetes of the young (MODY), gestational diabetes, non-alcoholic fatty liver disease (NAFLD), atherosclerosis, hypertension and coronary heart disease.
  • LADA latent immune diabetes in adults
  • MODY maturity-onset diabetes of the young
  • NAFLD non-alcoholic fatty liver disease
  • atherosclerosis hypertension and coronary heart disease.
  • the present patent application also relates to a method for agonizing the GLP-1 receptor, which comprises administering to a patient in need thereof a therapeutically effective amount of the compounds of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII) and specific compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same.
  • the present patent application also relates to a method for treating and/or preventing type I diabetes, type II diabetes, malnutrition-related diabetes, diabetic complications, obesity, hyperglycemia, glucose intolerance, cardiovascular disease, hyperlipidemia, cerebral infarction, stroke, non-alcoholic fatty liver disease (NASH), Parkinson's disease, dementia, insulin resistance and liver insulin resistance; preferably a method for type I diabetes, type II diabetes, obesity, diabetic complications, non-alcoholic fatty liver disease and cardiovascular disease, which comprises administering to a patient in need thereof a therapeutically effective amount of the general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (VI), general formula (VII) and specific compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same.
  • NASH non-alcoholic fatty liver disease
  • the present patent application also relates to a method for treating and/or preventing idiopathic type I diabetes, latent immune diabetes in adults (LADA), maturity-onset diabetes of the young (MODY), gestational diabetes, non-alcoholic fatty liver disease (NAFLD), atherosclerosis, hypertension and coronary heart disease, which comprises administering to a patient in need thereof a therapeutically effective amount of general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (VI), general formula (VII) and specific compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same.
  • LADA latent immune diabetes in adults
  • MODY maturity-onset diabetes of the young
  • NAFLD non-alcoholic fatty liver disease
  • atherosclerosis hypertension and coronary heart disease
  • the present patent application further relates to a general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (VI), general formula (VII) and specific compounds or their pharmaceutically acceptable salts, or pharmaceutical compositions containing the same, which are used as drugs.
  • This patent application also relates to a general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (VI), general formula (VII) and specific compounds, or a pharmaceutical composition containing the same, which is used as a GLP-1 receptor agonist.
  • This patent application further relates to a general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (V), Formula (VI), general formula (VII) and specific compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same, for treating and/or preventing type I diabetes, type II diabetes, malnutrition-related diabetes, diabetic complications, obesity, hyperglycemia, glucose intolerance, cardiovascular disease, hyperlipidemia, cerebral infarction, stroke, non-alcoholic steatohepatitis (NASH), Parkinson's disease, dementia, insulin resistance and liver insulin resistance; preferably for treating and/or preventing type I diabetes, type II diabetes, obesity, diabetic complications, non-alcoholic steatohepatitis and cardiovascular disease.
  • NASH non-alcoholic steatohepatitis
  • Parkinson's disease dementia
  • insulin resistance and liver insulin resistance preferably for treating and/or preventing type I diabetes, type II diabetes, obesity, diabetic complications, non-alcoholic steatohepatit
  • the present patent application further relates to a general formula (I), general formula (II), general formula (III), general formula (IV), general formula (V), general formula (VI), general formula (VII) and specific compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same, which are used to treat and/or prevent idiopathic type I diabetes, latent immune diabetes in adults (LADA), maturity-onset diabetes of the young (MODY), gestational diabetes, non-alcoholic fatty liver disease (NAFLD), atherosclerosis, hypertension and coronary heart disease.
  • idiopathic type I diabetes latent immune diabetes in adults (LADA), maturity-onset diabetes of the young (MODY), gestational diabetes, non-alcoholic fatty liver disease (NAFLD), atherosclerosis, hypertension and coronary heart disease.
  • LADA latent immune diabetes in adults
  • MODY maturity-onset diabetes of the young
  • NAFLD non-alcoholic fatty liver disease
  • atherosclerosis hypertension and coronary heart disease.
  • Diabetic complications are complications caused by diabetes or hyperglycemia, and they can be acute complexes or chronic complexes.
  • the term “acute complexes” includes ketoacidosis and infectious diseases (e.g., skin infections, soft tissue infections, biliary system infections, respiratory system infections, urinary tract infections), and "chronic complexes” include, for example, microangiopathy (e.g., nephropathy, retinopathy), neuropathy (e.g., sensory nerve disorders, motor nerve disorders, autonomic nerve disorders), and gangrene.
  • Major diabetic complexes include diabetic retinopathy, diabetic nephropathy, and diabetic neuropathy.
  • Chronic heart disease includes myocardial infarction and angina pectoris.
  • “Dementia” includes, for example, Alzheimer's disease, (early onset dementia) EOD, vascular dementia, and diabetic dementia.
  • the active compound can be prepared into a form suitable for administration by any appropriate route, and the composition of the present patent application can be prepared by conventional methods using one or more pharmaceutically acceptable carriers. Therefore, the active compound of the present patent application can be formulated into various dosage forms for oral administration, injection (e.g., intravenous, intramuscular or subcutaneous) administration, inhalation or insufflation administration.
  • injection e.g., intravenous, intramuscular or subcutaneous
  • inhalation or insufflation administration e.g., inhalation or insufflation administration.
  • the present invention provides a method for preparing the compound of formula I (Scheme 1),
  • PG 1 is an alkyl group or a benzyl group, such as a methyl group, an ethyl group, a tert-butyl group, etc., preferably a methyl group or an ethyl group;
  • PG 2 is an alkoxycarbonyl group, an alkanoyl group, an aroyl group, etc., for example, tert-butyloxycarbonyl, benzyloxycarbonyl, tert-methoxycarbonyl, allyloxycarbonyl, trimethylsilylethoxycarbonyl, trifluoroacetyl, phthaloyl, etc., preferably tert-butyloxycarbonyl;
  • LG 1 is halogen or substituted sulfonate, such as F, Cl, Br, I or triflate, preferably F, Cl or triflate;
  • LG 2 is halogen or substituted sulfonate, such as Cl, Br, I or triflate, preferably Br, I or triflate;
  • LG 3 is halogen or substituted sulfonate, such as Cl, Br, I or triflate, preferably Br, I;
  • LG 4 is a halogen, preferably Cl and Br;
  • reaction conditions of each step are as follows:
  • the first step the acyl halide prepared from raw material 1 undergoes nucleophilic substitution with raw material 2, or raw material 1 and raw material 2 directly
  • the amide intermediate 3 is then condensed.
  • the acyl halide reaction of the raw material 1 is carried out in a suitable organic solvent and a suitable halogenating agent, and the suitable organic solvent includes DCM, THF, DMF, etc., and the suitable halogenating agent includes oxalyl chloride, dichlorothionyl, etc.
  • the acyl halide prepared by the raw material 1 reacts with the raw material 2 in a nucleophilic substitution reaction in the presence of a suitable organic solvent, temperature, and base, and the suitable organic solvent includes DCM, THF, dioxane, etc., and the suitable temperature range is 0-25°C, and the suitable base includes triethylamine, TIPEA, N-methylmorpholine, etc.
  • the reaction of the raw material 1 and the raw material 2 condensing to form the amide intermediate 3 is carried out in the presence of a suitable condensing agent, and the suitable condensing agent includes HATU, PyBOP, DCC, etc.
  • Step 2 Deprotection of intermediate 3 to form intermediate 4.
  • a general deprotection scheme is selected according to the different protecting groups of PG 2. For example, when PG 2 is Boc, the Boc group is generally removed by acidic hydrolysis, and when PG 2 is Cbz, the Cbz group is removed by catalytic hydrogenolysis.
  • Step 3 Intermediate 4 undergoes intramolecular aromatic nucleophilic substitution to obtain isoindazolinone intermediate 5.
  • the reaction is carried out in a suitable organic solvent, including n-butanol, DMSO, etc., preferably DMSO; the reaction is carried out at a suitable reaction temperature, which is 120-170°C, such as 120°C, 130°C, etc., preferably 150°C.
  • Step 4 Intermediate 5 undergoes nucleophilic substitution with halide raw material 6 to obtain intermediate 7.
  • the reaction is carried out in a suitable organic solvent, including DMF, ethanol, etc., preferably DMF; the reaction is carried out at a suitable reaction temperature, 30-70°C, such as 30°C, 40°C, 50°C, etc., preferably 50°C.
  • the reaction is carried out in the presence of a suitable base, including Na 2 CO 3 , K 2 CO 3 , DIPEA, etc., preferably K 2 CO 3 ; preferably, the appropriate equivalent ratio of the reaction intermediate 5 to the intermediate 6 is 1:1-1:1.5.
  • the appropriate equivalent ratio of the intermediate 5 to the base is 1:1-1:3, preferably 1:2.
  • Step 5 Intermediate 7 is coupled with diborane pinacol ester to form borate ester compound 8.
  • the coupling reaction is preferably carried out in the presence of a metal catalyst and a base.
  • the metal catalyst is selected from a palladium metal catalyst, preferably [1,1′-bis(diphenylphosphino)ferrocene] palladium dichloride.
  • the base is preferably potassium acetate.
  • the coupling reaction preferably uses 1,4-dioxane as solvent.
  • the coupling reaction is carried out under a suitable inert atmosphere (e.g., a nitrogen environment).
  • the coupling reaction is carried out at a suitable temperature of 70 to 90° C., preferably 80° C.
  • the coupling reaction is carried out for a suitable time, such as 1 to 4 hours, such as 2 hours.
  • Step 6 Intermediate 8 and intermediate 9 undergo Suzuki coupling reaction to form linked aryl intermediate 10.
  • the coupling reaction is preferably carried out in the presence of a metal catalyst, an organophosphorus ligand and a base.
  • the metal catalyst is selected from a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride, triphenylphosphine palladium, palladium acetate, etc., preferably [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride.
  • the organophosphorus compound is selected from BINAP, Xantphos, X-Phos and S-Phos, etc.
  • the base is an inorganic base or an organic base, such as sodium tert-butoxide, potassium tert-butoxide, potassium phosphate, potassium carbonate, cesium carbonate, sodium carbonate, preferably cesium carbonate.
  • the coupling reaction is carried out in The coupling reaction is carried out in a mixed solvent consisting of a suitable organic solvent and water, and the mixed solvent can be selected from DMSO and water, DMF and water, 1,4-dioxane and water, toluene and water, xylene and water, etc.
  • the preferred mixed solvent is 1,4-dioxane and water, and the preferred volume ratio of the two is 10:1 to 5:1.
  • the coupling reaction is carried out under a suitable inert atmosphere (such as a nitrogen environment).
  • the coupling reaction is carried out at a suitable temperature, which can be, for example, 80 to 120°C, preferably 90°C or 100°C.
  • the coupling reaction is carried out for a suitable time, such as 8 to 24 hours, such as 12 hours.
  • Step 7 The ester intermediate 10 is subjected to alkaline hydrolysis to generate the final product 11.
  • the hydrolysis reaction is carried out in a suitable mixed solvent, the mixed solvent comprises water and another organic solvent, the organic solvent comprises methanol, ethanol, THF, etc., and the ratio of water to organic solvent in the mixed solvent is 4:1-1:4;
  • the reaction is carried out in the presence of a suitable base, the base comprises K 2 CO 3 , LiOH, TBD, etc., and the equivalent ratio of the intermediate 10 to the base is 1:2-1:3;
  • the reaction is carried out at a suitable reaction temperature, the reaction temperature is 40-60°C, such as 40°C, 50°C, etc., preferably 50°C.
  • the present invention provides a method for preparing a compound of formula VI, comprising the following steps:
  • PG 1 is an alkyl group or a benzyl group, such as a methyl group, an ethyl group, a tert-butyl group, etc., preferably a methyl group or an ethyl group;
  • PG 2 is an alkoxycarbonyl group, an alkanoyl group, an aroyl group, etc., for example, tert-butyloxycarbonyl, benzyloxycarbonyl, tert-methoxycarbonyl, allyloxycarbonyl, trimethylsilylethoxycarbonyl, trifluoroacetyl, phthaloyl, etc., preferably tert-butyloxycarbonyl;
  • LG 1 is halogen or substituted sulfonate, such as F, Cl, Br, I or triflate, preferably F, Cl or triflate;
  • LG 2 is halogen or substituted sulfonate, such as Cl, Br, I or triflate, preferably Br, I or triflate;
  • LG 3 is halogen or substituted sulfonate, such as Cl, Br, I or triflate, preferably Br, I;
  • LG 4 is a halogen, preferably Cl and Br;
  • reaction conditions of each step are as follows:
  • Step 1 The acyl halide prepared from raw material 1 undergoes nucleophilic substitution with raw material 2, or raw material 1 and raw material 2 are directly condensed to form amide intermediate 3.
  • the acyl halide preparation reaction of raw material 1 is carried out in a suitable organic solvent and a suitable halogenating agent, the suitable organic solvent includes DCM, THF, DMF, etc., and the suitable halogenating agent includes oxalyl chloride, dichlorothionyl, etc.
  • the acyl halide prepared from raw material 1 undergoes nucleophilic substitution with raw material 2 in the condensed state.
  • the reaction is carried out in the presence of a suitable organic solvent, temperature and base, wherein the suitable organic solvent includes DCM, THF, dioxane, etc., the suitable temperature range is 0-25° C., and the suitable base includes triethylamine, TIPEA, N-methylmorpholine, etc.
  • the reaction of condensing the raw material 1 with the raw material 2 to form the amide intermediate 3 is carried out in the presence of a suitable condensing agent, wherein the suitable condensing agent includes HATU, PyBOP, DCC, etc.
  • Step 2 Deprotection of intermediate 3 to form intermediate 4.
  • a general deprotection scheme is selected according to the different protecting groups of PG 2. For example, when PG 2 is Boc, the Boc group is generally removed by acidic hydrolysis, and when PG 2 is Cbz, the Cbz group is removed by catalytic hydrogenolysis.
  • Step 3 Intermediate 4 undergoes intramolecular aromatic nucleophilic substitution to obtain isoindazolinone intermediate 5.
  • the reaction is carried out in a suitable organic solvent, including n-butanol, DMSO, etc., preferably DMSO; the reaction is carried out at a suitable reaction temperature, which is 120-170°C, such as 120°C, 130°C, etc., preferably 150°C.
  • Step 4 Intermediate 5 undergoes nucleophilic substitution with raw material 6 to obtain intermediate 7.
  • the reaction is carried out in a suitable organic solvent, including DMF, ethanol, etc., preferably DMF; the reaction is carried out at a suitable reaction temperature, 30-70°C, such as 30°C, 40°C, 50°C, etc., preferably 50°C.
  • the reaction is carried out in the presence of a suitable base, including Na 2 CO 3 , K 2 CO 3 , DIPEA, etc., preferably K 2 CO 3 ; preferably, the suitable equivalent ratio of the reaction intermediate 5 to the intermediate 6 is 1:1-1:1.5.
  • the suitable equivalent ratio of the intermediate 5 to the base is 1:1-1:3, preferably 1:2.
  • Step 5 Intermediate 7 is coupled with diborane pinacol ester to form borate ester compound 8.
  • the coupling reaction is preferably carried out in the presence of a metal catalyst and a base.
  • the metal catalyst is selected from a palladium metal catalyst, preferably [1,1′-bis(diphenylphosphino)ferrocene] palladium dichloride.
  • the base is preferably potassium acetate.
  • the coupling reaction preferably uses 1,4-dioxane as solvent.
  • the coupling reaction is carried out under a suitable inert atmosphere (e.g., a nitrogen environment).
  • the coupling reaction is carried out at a suitable temperature of 70 to 90° C., preferably 80° C.
  • the coupling reaction is carried out for a suitable time, such as 1 to 4 hours, such as 2 hours.
  • Step 6 Intermediate 8 and intermediate 9 undergo Suzuki coupling reaction to form linked aryl intermediate 10.
  • the coupling reaction is preferably carried out in the presence of a metal catalyst, an organophosphorus ligand and a base.
  • the metal catalyst is selected from a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium, triphenylphosphine palladium, palladium acetate, etc., preferably [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium.
  • a palladium metal catalyst such as tris(dibenzylideneacetone)dipalladium, [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium, triphenylphosphine palladium, palladium acetate, etc., preferably [
  • the organophosphorus compound is selected from BINAP, Xantphos, X-Phos and S-Phos, etc.
  • the base is an inorganic base or an organic base, such as sodium tert-butoxide, potassium tert-butoxide, potassium phosphate, potassium carbonate, cesium carbonate, sodium carbonate, preferably cesium carbonate.
  • the coupling reaction is carried out in a mixed solvent consisting of a suitable organic solvent and water, and the mixed solvent can be selected from DMSO and water, DMF and water, 1,4-dioxane and water, toluene and water, xylene and water, etc.
  • the preferred mixed solvent is 1,4-dioxane and water, and the preferred volume ratio of the two is 10:1 to 5:1.
  • the coupling reaction is carried out in a suitable inert atmosphere (e.g. Preferably, the coupling reaction is carried out at a suitable temperature, which may be, for example, 80 to 120° C., preferably 90° C. or 100° C. Preferably, the coupling reaction is carried out for a suitable time, such as 8 to 24 hours, for example 12 hours.
  • Step 7 The ester intermediate 10 is subjected to alkaline hydrolysis to generate the final product 11.
  • the hydrolysis reaction is carried out in a suitable mixed solvent, the mixed solvent comprises water and another organic solvent, the organic solvent comprises methanol, ethanol, THF, etc., and the ratio of water to organic solvent in the mixed solvent is 4:1-1:4;
  • the reaction is carried out in the presence of a suitable base, the base comprises K 2 CO 3 , LiOH, TBD, etc., and the equivalent ratio of the intermediate 10 to the base is 1:2-1:3;
  • the reaction is carried out at a suitable reaction temperature, the reaction temperature is 40-60°C, such as 40°C, 50°C, etc., preferably 50°C.
  • Z 0 is CH
  • Z 1 is CR 13
  • Z 2 and Z 3 are N
  • Z 4 and Z 5 are both CH
  • X is O
  • Lp is CH 2
  • m 0
  • n 2
  • R 2 is Both R 3 are F and are located at the 2nd and 5th positions of the benzene ring respectively.
  • R 5 is When the following formula VII compound is obtained
  • the present invention provides a method for preparing a compound of Formula VII, comprising the following steps:
  • PG 1 is an alkyl group or a benzyl group, such as a methyl group, an ethyl group, a tert-butyl group, etc., preferably a methyl group or an ethyl group;
  • PG 2 is an alkoxycarbonyl group, an alkanoyl group, an aroyl group, etc., for example, tert-butyloxycarbonyl, benzyloxycarbonyl, tert-methoxycarbonyl, allyloxycarbonyl, trimethylsilylethoxycarbonyl, trifluoroacetyl, phthaloyl, etc., preferably tert-butyloxycarbonyl;
  • LG 1 is halogen or substituted sulfonate, such as F, Cl, Br, I or triflate, preferably F, Cl or triflate;
  • LG 2 is halogen or substituted sulfonate, such as Cl, Br, I or triflate, preferably Br, I or triflate;
  • LG 3 is halogen or substituted sulfonate, such as Cl, Br, I or triflate, preferably Br, I;
  • LG 4 is a halogen, preferably Cl and Br;
  • reaction conditions of each step are as follows:
  • Step 1 The acyl halide prepared from raw material 1 undergoes nucleophilic substitution with raw material 2, or raw material 1 and raw material 2 are directly condensed to form amide intermediate 3.
  • the acyl halide preparation reaction of raw material 1 is carried out in a suitable organic solvent and a suitable halogenating agent, the suitable organic solvent includes DCM, THF, DMF, etc., and the suitable halogenating agent includes oxalyl chloride, dichlorothionyl, etc.
  • the acyl halide prepared from raw material 1 undergoes nucleophilic substitution with raw material 2 in the condensed state.
  • the reaction is carried out in the presence of a suitable organic solvent, temperature and base, wherein the suitable organic solvent includes DCM, THF, dioxane, etc., the suitable temperature range is 0-25° C., and the suitable base includes triethylamine, TIPEA, N-methylmorpholine, etc.
  • the reaction of condensing the raw material 1 with the raw material 2 to form the amide intermediate 3 is carried out in the presence of a suitable condensing agent, wherein the suitable condensing agent includes HATU, PyBOP, DCC, etc.
  • Step 2 Deprotection of intermediate 3 to form intermediate 4.
  • a general deprotection scheme is selected according to the different protecting groups of PG 2. For example, when PG 2 is Boc, the Boc group is generally removed by acidic hydrolysis, and when PG 2 is Cbz, the Cbz group is removed by catalytic hydrogenolysis.
  • Step 3 Intermediate 4 undergoes intramolecular aromatic nucleophilic substitution to obtain isoindazolinone intermediate 5.
  • the reaction is carried out in a suitable organic solvent, including n-butanol, DMSO, etc., preferably DMSO; the reaction is carried out at a suitable reaction temperature, which is 120-170°C, such as 120°C, 130°C, etc., preferably 150°C.
  • Step 4 Intermediate 5 undergoes nucleophilic substitution with raw material 6 to obtain intermediate 7.
  • the reaction is carried out in a suitable organic solvent, including DMF, ethanol, etc., preferably DMF; the reaction is carried out at a suitable reaction temperature, 30-70°C, such as 30°C, 40°C, 50°C, etc., preferably 50°C.
  • the reaction is carried out in the presence of a suitable base, including Na 2 CO 3 , K 2 CO 3 , DIPEA, etc., preferably K 2 CO 3 ; preferably, the suitable equivalent ratio of the reaction intermediate 5 to the intermediate 6 is 1:1-1:1.5.
  • the suitable equivalent ratio of the intermediate 5 to the base is 1:1-1:3, preferably 1:2.
  • Step 5 Intermediate 7 is coupled with diborane pinacol ester to form borate ester compound 8.
  • the coupling reaction is preferably carried out in the presence of a metal catalyst and a base.
  • the metal catalyst is selected from a palladium metal catalyst, preferably [1,1′-bis(diphenylphosphino)ferrocene] palladium dichloride.
  • the base is preferably potassium acetate.
  • the coupling reaction preferably uses 1,4-dioxane as solvent.
  • the coupling reaction is carried out under a suitable inert atmosphere (e.g., a nitrogen environment).
  • the coupling reaction is carried out at a suitable temperature of 70 to 90° C., preferably 80° C.
  • the coupling reaction is carried out for a suitable time, such as 1 to 4 hours, such as 2 hours.
  • Step 6 Intermediate 8 and intermediate 9 undergo Suzuki coupling reaction to form linked aryl intermediate 10.
  • the coupling reaction is preferably carried out in the presence of a metal catalyst, an organophosphorus ligand and a base.
  • the metal catalyst is selected from a palladium metal catalyst, such as tris(dibenzylideneacetone)dipalladium, [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium, triphenylphosphine palladium, palladium acetate, etc., preferably [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium.
  • a palladium metal catalyst such as tris(dibenzylideneacetone)dipalladium, [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium, triphenylphosphine palladium, palladium acetate, etc., preferably [
  • the organophosphorus compound is selected from BINAP, Xantphos, X-Phos and S-Phos, etc.
  • the base is an inorganic base or an organic base, such as sodium tert-butoxide, potassium tert-butoxide, potassium phosphate, potassium carbonate, cesium carbonate, sodium carbonate, preferably cesium carbonate.
  • the coupling reaction is carried out in a mixed solvent consisting of a suitable organic solvent and water, and the mixed solvent can be selected from DMSO and water, DMF and water, 1,4-dioxane and water, toluene and water, xylene and water, etc.
  • the preferred mixed solvent is 1,4-dioxane and water, and the preferred volume ratio of the two is 10:1 to 5:1.
  • the coupling reaction is carried out in a suitable inert atmosphere (e.g. Preferably, the coupling reaction is carried out at a suitable temperature, which may be, for example, 80 to 120° C., preferably 90° C. or 100° C. Preferably, the coupling reaction is carried out for a suitable time, such as 8 to 24 hours, for example 12 hours.
  • Step 7 The ester intermediate 10 is subjected to alkaline hydrolysis to generate the final product 11.
  • the hydrolysis reaction is carried out in a suitable mixed solvent, the mixed solvent comprises water and another organic solvent, the organic solvent comprises methanol, ethanol, THF, etc., and the ratio of water to organic solvent in the mixed solvent is 4:1-1:4;
  • the reaction is carried out in the presence of a suitable base, the base comprises K 2 CO 3 , LiOH, TBD, etc., and the equivalent ratio of the intermediate 10 to the base is 1:2-1:3;
  • the reaction is carried out at a suitable reaction temperature, the reaction temperature is 40-60°C, such as 40°C, 50°C, etc., preferably 50°C.
  • the technical effect of the present invention is, on the one hand, to provide a compound with good GLP1R cell agonist activity.
  • the compound has an EC50 of less than 1000 pM in cell agonist activity.
  • the compound has an EC50 of less than 100 pM in cell agonist activity.
  • the compound has an EC50 of less than 80 pM in cell agonist activity.
  • the compound has an EC50 of less than 50 pM in cell agonist activity.
  • the compound has an EC50 of less than 20 pM in cell agonist activity.
  • the compound has an EC50 of less than 10 pM in cell agonist activity.
  • Another technical effect of the present invention is to provide a compound with good safety.
  • the compound of the present invention shows a weaker inhibitory effect than the compound of the prior art (such as PF06882961).
  • the compound of the present invention shows a higher hERG inhibition activity, for example, in some embodiments, the compound of the present invention shows that the inhibition IC 50 of hERG is greater than 30 ⁇ M. This shows that the compound of the present invention has better safety.
  • the technical effect of the present invention is on the other hand to provide a compound with good solubility.
  • the compound of the present invention shows a solubility far superior to that of existing compounds in a simulated fasting intestinal fluid solubility experiment.
  • the compound of the present invention has a solubility of 10-1000 ⁇ M.
  • the compound of the present invention has a solubility of 50-1000 ⁇ M.
  • the compound of the present invention has a solubility of 50-500 ⁇ M.
  • the compound of the present invention has a solubility of 100-500 ⁇ M.
  • Another technical effect of the present invention is to provide a compound with good bioavailability.
  • the compound of the present invention is significantly superior to the existing control compound in terms of oral exposure (Cmax and AUC) and bioavailability.
  • alkyl alkylene, alkenyl, “alkynyl”, “alkoxy”, "cycloalkyl”, “spirocycloalkyl”, “fused cycloalkyl”, “bridged cycloalkyl”, “fused heterocyclyl”, “aryl”, “heteroaryl” are all substituted or unsubstituted, and when substituted, they may be substituted at any available point of attachment.
  • alkyl refers to a saturated aliphatic hydrocarbon group having 1 to 20 carbon atoms (i.e., C1-20 alkyl).
  • the alkyl group preferably has 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12) carbon atoms (i.e., C1-12 alkyl), and more preferably has 1 to 6 carbon atoms (i.e., C1-6 alkyl).
  • 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,
  • lower alkyl groups having 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, it may be substituted at any available point of attachment.
  • the substituent is preferably selected from one or more of a D atom, a halogen, an alkoxy group, a haloalkyl group, a haloalkoxy group, a cycloalkyloxy group, a heterocyclyloxy group, a hydroxyl group, a hydroxyalkyl group, a cyano group, an amino group, a nitro group, a cycloalkyl group, a heterocyclyl group, an aryl group, and a heteroaryl group.
  • alkylene refers to a divalent alkyl group, wherein alkyl is as defined above, having 1 to 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) carbon atoms (i.e., C1-20 alkylene).
  • the alkylene group preferably has 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms (i.e., C1-12 alkylene), and more preferably has 1 to 6 carbon atoms (i.e., C1-6 alkylene).
  • alkylene groups include, but are not limited to, methylene (-CH2-), 1,1-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2)-, 1,1-propylene (-CH(CH2CH3)-), 1,2-propylene (-CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), 1,4-butylene (-CH2CH2CH2CH2-), and the like.
  • the alkylene group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment.
  • the substituents are preferably selected from one or more of alkenyl, alkynyl, alkoxy, haloalkoxy, cycloalkyloxy, heterocyclyloxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio and oxo.
  • alkenyl refers to an alkyl group containing at least one carbon-carbon double bond in a molecule, wherein the definition of alkyl is as described above, and it has 2 to 12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12) carbon atoms (i.e., C2-12 alkenyl).
  • the alkenyl group preferably has 2 to 6 carbon atoms (i.e., C2-6 alkenyl).
  • Non-limiting examples include: vinyl, propenyl, isopropenyl, butenyl, etc.
  • Alkenyl can be substituted or unsubstituted, and when substituted, it is preferably selected from one or more of alkoxy, halogen, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, and heteroaryl.
  • alkynyl refers to an alkyl group containing at least one carbon-carbon triple bond in the molecule, wherein the definition of alkyl is as described above, and it has 2 to 12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms.
  • the alkynyl group preferably has 2 to 6 carbon atoms (i.e., C2-6 alkynyl).
  • the alkynyl group can be substituted or unsubstituted, and when substituted, it is preferably selected from one or more of alkoxy, halogen, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.
  • alkoxy refers to -O-(alkyl), wherein alkyl is as defined above. Non-limiting examples include: methoxy, ethoxy, propoxy and butoxy, etc.
  • Alkoxy can be substituted or unsubstituted, and when substituted, it can be substituted at any available point of attachment, and the substituent is preferably selected from one or more of a deuterium atom, a halogen, an alkoxy, a haloalkyl, a haloalkoxy, a cycloalkyloxy, a heterocyclyloxy, a hydroxyl, a hydroxyalkyl, a cyano, an amino, a nitro, a cycloalkyl, a heterocyclyl, an aryl and a heteroaryl.
  • cycloalkyl refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein the cycloalkyl ring has 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) carbon atoms, preferably 3 to 12 carbon atoms (i.e., 3 to 12-membered cycloalkyl), more preferably 3 to 8 (e.g., 3, 4, 5, 6, 7 and 8) carbon atoms (i.e., 3 to 8-membered cycloalkyl), and most preferably 3 to 6 carbon atoms (i.e., 3 to 6-membered cycloalkyl).
  • 3 to 20 e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20
  • 3 to 12 carbon atoms i.e., 3 to 12-membered cycloalkyl
  • 3 to 8 e.g., 3, 4, 5, 6, 7 and
  • Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl and cyclooctyl, etc.; polycyclic cycloalkyls include spirocycloalkyls, fused cycloalkyls and bridged cycloalkyls.
  • spirocycloalkyl refers to a polycyclic group of 5 to 20 yuan, in which a carbon atom (called a spiral atom) is shared between the monocyclic rings, and it may contain one or more double bonds. Preferably, it is 6 to 14 yuan, and more preferably, it is 7 to 10 yuan (e.g., 7, 8, 9 or 10 yuan).
  • the spirocycloalkyl is divided into a single spiral cycloalkyl or a multi-spirocycloalkyl (e.g., a double spiral cycloalkyl), preferably a single spiral cycloalkyl or a double spiral cycloalkyl. 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 spiral cycloalkyl.
  • condensed cycloalkyl refers to 5 to 20 yuan, and each ring in the system shares a pair of adjacent carbon atoms with other rings in the system. All carbon polycyclic groups, wherein one or more rings can contain one or more double bonds. Preferably 6 to 14 yuan, more preferably 7 to 10 yuan (such as 7,8,9 or 10 yuan).
  • the polycyclic condensed cycloalkyls such as bicyclic, tricyclic, tetracyclic can be divided, preferably bicyclic or tricyclic condensed cycloalkyl, more preferably 3 yuan/4 yuan, 3 yuan/5 yuan, 3 yuan/6 yuan, 4 yuan/4 yuan, 4 yuan/5 yuan, 4 yuan/6 yuan, 5 yuan/4 yuan, 5 yuan/3 yuan, 5 yuan/5 yuan, 5 yuan/6 yuan, 6 yuan/3 yuan, 6 yuan/4 yuan, 6 yuan/5 yuan, 6 yuan/6 yuan, 6 yuan/7 yuan, 7 yuan/5 yuan or 7 yuan/6 yuan of bicycl
  • bridged cycloalkyl refers to a 5 to 20-membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, and which may contain one or more double bonds. Preferably, it is 6 to 14 members, and more preferably, it is 7 to 10 members (e.g., 7, 8, 9 or 10 members). According to the number of constituent rings, it can be divided into polycyclic bridged cycloalkyls such as bicyclic, tricyclic, and tetracyclic, preferably bicyclic, tricyclic or tetracyclic bridged cycloalkyls, and more preferably bicyclic or tricyclic bridged cycloalkyls.
  • the cycloalkyl ring includes the cycloalkyl group as described above (including monocyclic, spirocyclic, condensed and bridged rings) fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group.
  • the cycloalkyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably selected from one or more of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.
  • heterocyclyl refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic substituent having 3 to 20 ring atoms, one or more of which are heteroatoms selected from nitrogen, oxygen and sulfur, the sulfur being optionally oxidized (i.e. forming sulfoxide or sulfone), and the remaining ring atoms being carbon.
  • it has 3 to 12 (e.g. 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) ring atoms, of which 1 to 4 (e.g. 1, 2, 3 and 4) are heteroatoms (i.e. 3 to 12-membered heterocyclyl); more preferably, it has 3 to 8 (e.g.
  • ring atoms of which 1 to 3 (e.g. 1, 2 and 3) are heteroatoms (i.e. 3 to 8-membered heterocyclyl); more preferably, it has 3 to 6 ring atoms, of which 1 to 3 are heteroatoms (i.e. 3 to 6-membered heterocyclyl); most preferably, it has 5 or 6 ring atoms, of which 1 to 3 are heteroatoms (i.e. 5 or 6-membered heterocyclyl).
  • 1 to 3 e.g. 1, 2 and 3
  • heteroatoms i.e. 3 to 8-membered heterocyclyl
  • 3 to 6 ring atoms of which 1 to 3 are heteroatoms (i.e. 3 to 6-membered heterocyclyl)
  • Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, tetrahydropyranyl, 1,2,3,6-tetrahydropyridinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc.
  • Polycyclic heterocyclyls include spiroheterocyclyls, fused heterocyclyls, and bridged heterocyclyls.
  • spiroheterocyclyl refers to a polycyclic heterocyclic group of 5 to 20 members, in which one atom (called spiro atom) is shared between single rings, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, the sulfur may be optionally oxoed (i.e. forming sulfoxide or sulfone), and the remaining ring atoms are carbon. It may contain one or more double bonds.
  • 6 to 14 members more preferably 7 to 10 members (e.g. 7, 8, 9 or 10 members).
  • spiroheterocyclyl is divided into monospiroheterocyclyl or polyspiroheterocyclyl (e.g. bispiroheterocyclyl), preferably monospiroheterocyclyl or bispiroheterocyclyl.
  • fused heterocyclic group refers to a polycyclic heterocyclic group of 5 to 20 members, 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, one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, the sulfur may be optionally oxoed (i.e., forming sulfoxide or sulfone), and the remaining ring atoms are carbon.
  • 6 to 14 members more preferably 7 to 10 members (e.g., 7, 8, 9 or 10 members).
  • the number of constituent rings it can be divided into bicyclic, tricyclic, tetracyclic and other polycyclic fused heterocyclic groups, preferably a bicyclic or tricyclic fused heterocyclic group, more preferably a 3-membered/4-membered, 3-membered/5-membered, 3-membered/6-membered, 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/3-membered, 5-membered/4-membered, 5-membered/5-membered, 5-membered/6-membered, 6-membered/3-membered, 6-membered/4-membered, 6-membered/5-membered, 6-membered/6-membered, 6-membered/7-membered, 7-membered/5-membered or 7-membered/6-membered bicyclic fused heterocyclic group.
  • bridged heterocyclic group refers to a polycyclic heterocyclic group of 5 to 14 members, any two rings sharing two atoms that are not directly connected, which may contain one or more double bonds, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, the sulfur may be optionally oxoed (i.e., forming sulfoxide or sulfone), and the remaining ring atoms are carbon.
  • 6 to 14 members more preferably 7 to 10 members (e.g., 7, 8, 9 or 10 members).
  • the number of constituent rings it can be divided into bicyclic, tricyclic, tetracyclic and other polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic bridged heterocyclic groups, more preferably bicyclic or tricyclic bridged heterocyclic groups.
  • the heterocyclic ring includes a heterocyclic group as described above (including a monocyclic, spiro heterocyclic, fused heterocyclic and bridged heterocyclic) fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, and the heterocyclic group may be substituted or unsubstituted.
  • the substituent is preferably selected from one or more of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclic groupoxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic group, aryl and heteroaryl.
  • aryl refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (fused polycyclic is a ring that shares adjacent carbon atom pairs) group with a conjugated ⁇ electron system, preferably 6- to 10-membered, such as phenyl and naphthyl.
  • the aryl ring includes an aryl ring as described above fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring.
  • the aryl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably selected from one or more of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.
  • heteroaryl refers to a heteroaromatic system containing 1 to 4 (e.g., 1, 2, 3, and 4) heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen.
  • the heteroaryl group is preferably 5 to 10 members (e.g., 5, 6, 7, 8, 9, or 10 members), more preferably 5 or 6 members (i.e., 5 or 6 membered heteroaryl), such as furanyl, thienyl, pyridyl, Pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, etc.
  • the heteroaryl ring includes a heteroaryl fused to an aryl, heterocyclic or cycloalkyl ring as described above, wherein the ring connected to the parent structure is a heteroaryl ring.
  • the heteroaryl group can be substituted or unsubstituted, and when substituted, it can be substituted at any available point of attachment, and the substituent is preferably selected from one or more of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclic oxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, and heteroaryl.
  • cycloalkyl, heterocyclyl, aryl and heteroaryl groups include residues derived from a parent ring atom by removing one hydrogen atom, or residues derived from the same or two different ring atoms of the parent by removing two hydrogen atoms, i.e., "cycloalkylene", “heterocyclylene”, “arylene” and "heteroarylene”.
  • amino protecting group refers to a group that is easily removed and introduced on the amino group in order to keep the amino group unchanged when other parts of the molecule react.
  • Non-limiting examples include (trimethylsilyl)ethoxymethyl, tetrahydropyranyl, tert-butyloxycarbonyl (Boc), acetyl, benzyl, benzyloxycarbonyl (Cbz), allyl and p-methoxybenzyl, etc.
  • These groups can be optionally substituted with 1-3 substituents selected from halogen, alkoxy or nitro.
  • hydroxy protecting group is a suitable group for hydroxy protecting known in the art.
  • Non-limiting examples include: trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), tert-butyldimethylsilyl (TBS), tert-butyldiphenylsilyl, methyl, tert-butyl, allyl, benzyl, methoxymethyl (MOM), ethoxyethyl, 2-tetrahydropyranyl (THP), formyl, acetyl, benzoyl, p-nitrobenzoyl, etc.
  • cycloalkyloxy refers to a cycloalkyl-O- group wherein cycloalkyl is as defined above.
  • heterocyclyloxy refers to heterocyclyl-O-, wherein heterocyclyl is as defined above.
  • alkylthio refers to an alkyl-S- group in which alkyl is as defined above.
  • haloalkyl refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.
  • haloalkoxy refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.
  • deuterated alkyl refers to an alkyl group substituted with one or more deuterium atoms, wherein alkyl is as defined above.
  • hydroxyalkyl refers to an alkyl group substituted with one or more hydroxy groups, wherein alkyl is as defined above.
  • halogen refers to fluorine, chlorine, bromine or iodine.
  • hydroxy refers to -OH.
  • thiol refers to -SH.
  • amino refers to -NH2.
  • cyano refers to -CN.
  • nitro refers to -NO2.
  • carboxylate refers to -C(O)O(alkyl), -C(O)O(cycloalkyl), (alkyl)C(O)O-, or (cycloalkyl)C(O)O-, wherein alkyl and cycloalkyl are as defined above.
  • the compounds of this patent application include isotopic derivatives thereof.
  • isotopic derivative refers to a compound whose structure differs only in the presence of one or more isotopically enriched atoms.
  • compounds having the structure of this patent application replacing hydrogen with “deuterium” or “tritium”, or replacing fluorine with 18F-fluorine labeling (18F isotope), or replacing carbon atoms with 11C-, 13C-, or 14C-enriched carbon (11C-, 13C-, or 14C-carbon labeling; 11C-, 13C-, or 14C-isotope) are within the scope of this patent application.
  • Such compounds can be used as analytical tools or probes in, for example, biological assays, or can be used as in vivo diagnostic imaging tracers for diseases, or as tracers for pharmacodynamics, pharmacokinetics, or receptor studies.
  • the various deuterated forms of compounds of this patent application refer to compounds in which each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize deuterated forms of compounds with reference to relevant literature.
  • deuterated starting materials can be used to prepare deuterated compounds, or they can be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated borane tetrahydrofuran solution, deuterated lithium aluminum hydride, deuterated iodoethane and deuterated iodomethane, etc.
  • Deuterated compounds can generally retain activities comparable to those of undeuterated compounds, and when deuterated at certain specific sites, better metabolic stability can be achieved, thereby obtaining certain therapeutic advantages.
  • stereoisomer refers to isomers with the same structure but different arrangements of atoms in space. It includes cis and trans (or Z and E) isomers, (-)- and (+)-isomers, (R)- and (S)-enantiomers, diastereomers, (D)- and (L)-isomers, tautomers, atropisomers, conformers and mixtures thereof (such as racemates, mixtures of diastereomers).
  • the substituents in the compounds of this patent application may have additional asymmetric atoms. All of these stereoisomers and their mixtures are included within the scope of this patent application.
  • Optically active (-)- and (+)-isomers, (R)- and (S)-enantiomers and (D)- and (L)-isomers may be prepared by chiral synthesis, chiral reagents or other conventional techniques.
  • An isomer of a compound of the present patent application can be prepared by asymmetric synthesis or chiral auxiliary, or, when the molecule contains a basic functional group (such as amino group) or an acidic functional group (such as carboxyl group), a diastereoisomer salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art to obtain pure isomers.
  • the separation of enantiomers and diastereomers is usually completed by chromatography.
  • the bond "/" represents an unspecified configuration, that is, if chiral isomers exist in the chemical structure, the bond “/" can be or or contain both and configurations.
  • tautomer or tautomeric form refers to a tautomer that exists in equilibrium and Structural isomers that are easily converted from one isomeric form to another isomeric form. It includes all possible tautomers, i.e., in the form of a single isomer or in the form of a mixture of any proportions of the tautomers. Non-limiting examples include: keto-enol, imine-enamine, lactam-lactim, etc. All tautomeric forms are within the scope of this patent application, and the naming of the compound does not exclude any tautomer.
  • 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 or does not occur.
  • a heterocyclic group optionally substituted with an alkyl group means that an alkyl group may but need not be present, and the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.
  • Substituted means that one or more hydrogen atoms, preferably 1 to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently by a corresponding number of substituents.
  • substituents Those skilled in the art can determine possible or impossible substitutions (by experiment or theory) without undue effort.
  • an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.
  • “Pharmaceutical composition” means a mixture containing one or more compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients.
  • the purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.
  • “Pharmaceutically acceptable salts” refer to salts of the compounds of the present application, which are safe and effective when used in mammals and have the desired biological activity. Salts can be prepared separately during the final isolation and purification of the compound, or by reacting a suitable group with a suitable base or acid.
  • Bases commonly used to form pharmaceutically acceptable salts include inorganic bases and organic bases.
  • Acids commonly used to form pharmaceutically acceptable salts include inorganic acids and organic acids.
  • the term "therapeutically effective amount” refers to an amount of the drug or agent sufficient to achieve or at least partially achieve the desired effect.
  • the determination of a therapeutically effective amount varies from person to person, depending on the age and general condition of the recipient and on the specific active substance, and the appropriate therapeutically effective amount in each case can be determined by a person skilled in the art based on routine experiments.
  • pharmaceutically acceptable refers to those compounds, materials, compositions and/or dosage forms that are, within the scope of sound medical judgment, suitable for contact with patient tissues without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit/risk ratio, and effective for the intended use.
  • FIG. 1 Blood glucose-time variation curves of each test compound in Example 52
  • FIG. 2 Comparison of blood glucose-time integral (AUC) of each test compound in Example 52
  • FIG. 3 Random blood glucose-time variation curves of each test compound in Example 53
  • FIG. 4 Fasting blood glucose-time change curves of each test compound in Example 53
  • FIG. 5 Blood glucose-time variation curves of each test compound in Example 53
  • the 1 H-NMR was measured by a Bruker 400 MHz nuclear magnetic resonance spectrometer, the measuring solvent was deuterated methanol (CD 3 OD), deuterated chloroform (CDCl 3 ) or hexadeuterated dimethyl sulfoxide (DMSO-d6), and the internal standard was tetramethylsilane (TMS).
  • TMS tetramethylsilane
  • MS mass spectrometry
  • ESI Agilent
  • TLC Thin layer chromatography
  • the reaction is monitored by thin layer chromatography (TLC) or liquid chromatography-mass spectrometry (LC-MS), and the developing solvent systems used include dichloromethane and methanol system, n-hexane and ethyl acetate system, and petroleum ether and ethyl acetate system.
  • the developing solvent system is adjusted according to the polarity of the compound to be separated (by adjusting the volume ratio of the solvent or adding triethylamine, etc.).
  • the instrument model used for preparative high performance liquid chromatography is: Agilent 1260, chromatographic column: Waters XBridge Prep C18OBD (19mm ⁇ 150mm ⁇ 5.0 ⁇ m); column temperature: 25°C; flow rate: 20.0mL/min; detection wavelength: 214nm; elution gradient: (0min: 10% A, 90% B; 16.0min: 90% A, 10% B); mobile phase A: 100% acetonitrile; mobile phase B: 0.05% ammonium bicarbonate aqueous solution.
  • reaction temperature is room temperature (20°C to 30°C).
  • the reagents used in the examples were purchased from Acros Organics, Aldrich Chemical Company, Shanghai Tebo Chemical Technology Co., Ltd., etc.
  • Step 1 Add potassium carbonate (14.50 g, 104.89 mmol) to a DMF (100 mL) solution of raw material Int 1b1 (10 g, 69.93 mmol) and bromoacetaldehyde diethyl acetal (16.55 g, 83.92 mmol), and stir at 120 ° C overnight. After the reaction is completed by TLC monitoring, 100 mL of water is added, and ethyl acetate (100 mL x 3) is extracted. The organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate is concentrated to obtain the crude product Int 1b2, which is used directly in the next step without purification.
  • Step 2 Add 30 mL of PPA to a solution of intermediate Int 1b2 (18 g, 69.56 mmol) in DCE (180 mL), and stir overnight at 80°C under nitrogen protection. After the reaction is completed by TLC monitoring, 200 mL of water is added, and the mixture is extracted with ethyl acetate (200 mL x 3). The organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the concentrated filtrate is purified by silica gel column chromatography to obtain Int 1b3.
  • Step 3 Add NBS (10.00 g, 56.04 mmol) and AIBN (1.54 g, 9.34 mmol) to a DCE (80 mL) solution of the intermediate Int 1b3 (7.8 g, 46.70 mmol), heat to 72 °C and stir overnight. After the reaction is complete, add 100 mL of water, extract with ethyl acetate (100 mL x 3), wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, and purify the concentrated filtrate with a silica gel column to obtain Int 1b4.
  • Step 4 Add potassium acetate (8 g, 81.30 mmol) to a DMF (20 mL) solution of the intermediate Int 1b4 and stir at room temperature for 3 h. After the reaction is completed by TLC monitoring, 20 mL of water is added and extracted with ethyl acetate (20 mL x 3). The organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated to obtain the crude product Int 1b5, which is used directly in the next step without purification.
  • Step 5 Sodium methoxide (0.86 g, 16.00 mmol) was added to a solution of intermediate Int 1b5 (1.8 g, 8.00 mmol) in THF (20 mL), and the mixture was stirred at room temperature for 1 h. After the reaction was completed, 100 mL of water was added, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the concentrated filtrate was purified by silica gel column chromatography. Spectral purification afforded Int 1b6.
  • Step 7 Add LDA (2M, 2.2 mL, 4.40 mmol) to a solution of intermediate Int 1b7 (1.0 g, 2.95 mmol) in ultra-dry tetrahydrofuran (10 mL), stir at -60 ° C for 1 h, then add N-fluorobisbenzenesulfonamide (9.3 g, 29.50 mmol), continue stirring at -60 ° C for 1 h, warm to room temperature and stir for another 1 h.
  • Step 1 Add 1,1-difluoro-2-iodoethane (1.1 g, 5.73 mmol) and cesium carbonate (3.7 g, 11.35 mmol) to a DMF (10 mL) solution of the raw material Int 1c-1 (1 g, 5.68 mmol), stir at room temperature for 16 h, and after the reaction is complete, concentrate.
  • Step 2 Methanol (194 mg, 6.06 mmol) and lithium borohydride (3 mL, 6.06 mmol) were added to a solution of intermediate Int 1c-2 (725 mg, 3.02 mmol) in THF (8 mL), and the mixture was stirred at 50 ° C for 12 h. After the reaction was completed, saturated ammonium chloride (10 mL) was added to quench, ethyl acetate (15 mL x 3) was extracted, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain crude product Int 1c-3.
  • LCMS (ESI, m/z): 213.0 [M+H] + .
  • Step 3 Add 2-fluoro-6-bromopyridine (608 mg, 3.45 mmol) and potassium tert-butoxide (484 mg, 4.32 mmol) in tetrahydrofuran (3 mL) to a solution of intermediate Int 1c-3 (733 mg, 3.46 mmol) in dioxane (10 mL), and stir the reaction at 40 ° C for 30 min. After the reaction is completed, add saturated ammonium chloride (20 mL) solution to quench, add water (20 mL) to dilute the reaction system, extract with ethyl acetate (15 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain crude Int 1c. LCMS (ESI, m/z): 369.9.
  • Step 1 Add dimethyl sulfide borane (16 mL, 32.17 mmol) to a solution of raw material Int 1e1 (950 mg, 5.36 mmol) in THF (10 mL) and stir at room temperature for 1 h. After the reaction is complete, add water (10 mL) and ethyl acetate. The product was extracted with ester (10 mL x 3), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the intermediate Int 1e2. The crude product was used directly in the next step.
  • Step 2 Add t-BuOK (420.51 mg, 3.75 mmol) in THF (2 mL) to a solution of intermediate Int 1e2 (510 mg, 3.13 mmol) and 2-fluoro-6-bromopyridine (550.88 mg, 3.13 mmol) in dioxane (6 mL), and stir at 45 °C for 0.5 h. After the reaction is complete, add water (10 mL), extract with ethyl acetate (10 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify with a silica gel column to obtain Int 1e. LCMS (ESI, m/z): 318.9 [M+H] + .
  • Step 1 Add imidazole (5 g, 73.44 mmol) to a DCM (100 mL) solution of the raw material Int 1f1 (5.0 g, 48.95 mmol) at 0°C, add dropwise a DCM (40 mL) solution of TBSCl (7.35 g, 48.95 mmol), and stir at room temperature for 3 hours. After the reaction is completed, cool to 0°C, add water (50 ml), extract with ethyl acetate (120 mL x 3), dry with sodium sulfate, concentrate, and purify the crude product with a silica gel column to obtain Int 1f2.
  • Step 2 Add triphenylphosphine (6.24 g, 23.78 mmol), imidazole (1.62 g, 23.78 mmol) and iodine (5.54 g, 21.79 mmol) to a DMF (100 mL) solution of the intermediate Int 1f2 (4.3 g, 19.81 mmol), stir at -20°C for 2 h, add water (200 ml) after the reaction, extract with ether (120 mL x 3), dry the organic phase with sodium sulfate, concentrate, and purify the crude product on a silica gel column to obtain Int 1f.
  • Step 1 Add dimethylamine tetrahydrofuran solution (8.5 mL) to the raw material Int 1f-5-1 (850 mg, 5.04 mmol) and stir at 70°C for 12 h. After the reaction is completed, concentrate. Purify by silica gel column to obtain Int 1f-5-2. LCMS (ESI, m/z): 214.0 [M+H] + .
  • Step 2 To a solution of intermediate Int 1f-5-2 (600 mg, 2.80 mmol) and 2-fluoro-6-bromopyridine (493.46 mg, 2.80 mmol) in dioxane (6 mL), add a solution of t-BuOK (392.52 mg, 3.50 mmol) in THF (2 mL), and stir at 45 ° C for 0.5 h. After the reaction is completed, water (10 mL) is added, and ethyl acetate (10 mL x 3) is extracted, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column to obtain Int 1f-5.
  • t-BuOK 392.52 mg, 3.50 mmol
  • Step 1 To a solution of 4-bromo-2,5-difluorobenzaldehyde Int 2a-1 (5 g, 22.7 mmol) in THF (50 mL), add tert-butyl carbazate (3.6 g, 27.2 mmol) and sodium cyanoborohydride (2.8 g, 45.4 mmol). Stir at 50 °C for 3 h. After the reaction is complete, concentrate the reaction solution and use the crude product Int 2a-2 directly in the next step.
  • Step 2 Sodium borohydride (1.7 g, 45.4 mmol) was added to a methanol (60 mL) solution of the intermediate Int 2a-2 (7.6 g, 22.7 mmol), and the reaction was stirred at room temperature for 1 h. After the reaction was completed, water (100 mL) was added to dilute the reaction system, and ethyl acetate (10 mL x 3) was used for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography to obtain Int 2a. LCMS (ESI, m/z): 378.2 [M+H+MeCN] + .
  • Step 1 Add borane dimethyl sulfide (21.4 mL, 2M) to a THF (50 mL) solution of the raw material Int 2c-1 (5 g, 21.46 mmol) and stir at room temperature overnight. After the reaction is complete, add HCl (1N, 10 mL) to quench the reaction, filter the reaction solution, concentrate the filtrate to remove tetrahydrofuran, add water (30 mL), extract with ethyl acetate (3 ⁇ 50 mL), dry the organic phase, filter, and evaporate the solvent under reduced pressure to obtain the crude product of Int 2c-2.
  • 1 H NMR 400 MHz, CDCl 3 ) ⁇ 7.16-7.23 (m, 2H), 4.61 (s, 2H), 2.29 (s, 3H).
  • Step 2 The intermediate Int 2c-2 (4.3 g, 19.63 mmol) was dissolved in dichloromethane (100 mL), and Dess-Martin periodinane (16.65 g, 39.26 mmol) was added at 0°C, and stirred at room temperature overnight. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate solution (80 mL), extracted with dichloromethane (3 ⁇ 100 mL), and the organic phase was washed with 10% sodium thiosulfate solution, dried over sodium sulfate, filtered, concentrated to remove the solvent, and the crude product was purified by silica gel column chromatography to obtain Int2c-3.
  • Step 3 The intermediate Int 2c-3 (3.187 g, 14.68 mmol) and tert-butyloxycarbonylhydrazine (2.91 g, 22.03 mmol) were dissolved in methanol (50 mL) and reacted at 50°C for 2 h. After the reaction, the solvent was evaporated under reduced pressure and the crude product was purified by silica gel column chromatography to obtain Int 2c-4. LCMS (ESI, m/z): 275.0 [M+H-56] + .
  • Step 4 The intermediate Int 2c-4 (4.70 g, 14.20 mmol) was dissolved in THF (80 mL), and a tetrahydrofuran solution of DIBAL-H (59.4 mL, 1.5 M) was added dropwise at -78 °C under nitrogen protection. After the addition, the temperature was gradually raised to room temperature for 2 h. After the reaction, methanol (10 mL) was added dropwise to quench the reaction. The reaction solution was concentrated, and a saturated potassium sodium tartrate solution (300 mL) was added to the residue, stirred until the solution was clear, and then ethyl acetate (200 mL ⁇ 3) was added for extraction.
  • DIBAL-H 59.4 mL, 1.5 M
  • Step 1 Add tert-butyl carbazate to a solution of the raw material Int 2d-1 (5 g, 24.63 mmol) in tetrahydrofuran (50 mL), and react at 50°C for 2 h. After the reaction is complete, the reaction solution is returned to room temperature and concentrated under vacuum to obtain Int 2d-2.
  • LCMS (ESI, m/z): 278.9 [M-55] + .
  • Step 2 Under nitrogen protection, a solution of the intermediate Int 2d-2 (9 g, 26.85 mmol) in tetrahydrofuran (100 mL) was cooled to -78°C, and DIBAL-H (100 mL, 161.12 mmol) was added and reacted at -78°C for 1 h. After the reaction was complete, the reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain a crude product, which was purified by silica gel column chromatography to obtain Int 2d. LCMS (ESI, m/z): 281.0 [M-55] + .
  • Step 1 Dissolve the raw materials trimethyl sulfoxide iodide (161 g, 732 mmol) and potassium tert-butoxide (82 g, 732 mmol) in tert-butanol (600 mL) solution, stir at 60 ° C for 0.5 h, then heat to 80 ° C, dropwise add the raw material Int 3-1 (60 g, 366 mmol), and stir for 2 h.
  • Step 2 Add palladium carbon (10 g) to a solution of intermediate Int 3-2 (25 g, 140.4 mmol) in methanol (350 mL), and stir at 50 ° C for 12 h under a hydrogen atmosphere. After the reaction is completed, filter, concentrate the filtrate, and purify it with a silica gel column to obtain Int 3-3.
  • Step 3 Add imidazole (12.85 g, 189 mmol), triphenylphosphine (24.65 g, 94.1 mmol) and iodine (23.9 g, 94.1 mmol) to 50 mL of dichloromethane and stir at 0 ° C for 0.5 h. After returning to room temperature, add intermediate Int 3-3 (6.44 g, 73.1 mmol) dropwise and stir at room temperature for 12 h.
  • Example 1 Synthesis of 2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-3-(2-methoxyethyl)-3-methyl-1-oxoisoindoline-5-carboxylic acid (KH01), (R)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-3-(2-methoxyethyl)-3-methyl-1-oxoisoindoline-5-carboxylic acid (KH01-1) and (S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2-fluorobenzyl)-3-(2-methoxyethyl)-3-methyl-1-oxoisoindoline-5-carboxylic acid (KH01-2)
  • Step 1 Add p-methoxybenzylamine (2.23 g, 16.23 mmol) and triethylamine (3.28 g, 32.46 mmol) to a DMF (50 mL) solution of methyl 2-bromomethyl-4-bromobenzoate KH 1-1 (5 g, 16.23 mmol), respectively, and stir at room temperature for 2 h. After the reaction is completed, add 100 mL of water, extract with EA (100 mL*3), wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, and separate the concentrated filtrate by silica gel column chromatography to obtain KH 1-2.
  • Step 2 Sodium hydride (362 mg, 9.06 mmol) was added to a DMF (15 mL) solution of KH 1-2 (1.5 g, 4.53 mmol). The mixture was stirred at 80 °C for 30 min, cooled to room temperature, and iodomethane (772 mg, 5.44 mmol) was added to the mixture, and stirred at room temperature for 1 h. After the reaction was completed, 15 mL of water was added, and EA was extracted (15 mL ⁇ 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the concentrated crude product was purified by silica gel column chromatography to obtain KH1-3. LCMS (ESI, m/z): 346.0 [M+H] + .
  • Step 3 Sodium hydride (158 mg, 3.94 mmol) and 2-bromoethyl methyl ether (544 mg, 3.94 mmol) were added to a DMF (7 mL) solution of KH 1-3 (680 mg, 1.97 mmol), and the mixture was stirred at 80°C for 1 h. After the reaction was completed, water (10 mL) was added, and EA (10 mL*3) was used for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography to obtain KH 1-4.
  • Step 4 Pd(dppf)Cl 2 (88 mg, 0.12 mmol) and triethylamine (627 mg, 6.21 mmol) were added to a solution of KH1-4 (500 mg, 1.24 mmol) in ethanol (10 mL), and stirred at 120°C for 16 h under a carbon monoxide atmosphere (four standard atmospheres). After the reaction was completed, the solvent was evaporated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain KH 1-5.
  • Step 5 A solution of KH 1-5 (380 mg, 0.96 mmol) in trifluoroacetic acid (5 mL) was stirred at 80° C. for 16 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain KH 1-6.
  • Step 6 To a solution of KH 1-6 (200 mg, 0.72 mmol) in DMF (5 mL) was added sodium hydride (58 mg, 1.44mmol), the mixture was stirred at room temperature for 30min, 4-bromo-2-fluorobenzyl bromide (232mg, 0.86mmol) was added to the mixture, and the mixture was stirred at room temperature for 16h. After the reaction was completed, water (10mL) was added, and EA (10mL ⁇ 3) was extracted. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The crude product of the concentrated filtrate was purified by silica gel column chromatography to obtain KH1-7. LCMS (ESI, m/z): 464.0[M+H] + .
  • Step 7 Add pinacol diboronate (248 mg, 0.97 mmol), Pd(dppf)Cl 2 (44 mg, 0.06 mmol) and potassium acetate (128 mg, 1.31 mmol) to a solution of KH 1-7 (300 mg, 0.65 mmol) in dioxane (5 mL), respectively, and stir the mixture at 80°C for 4 h. After the reaction, the crude product was used directly in the next step without further treatment. LCMS (ESI, m/z): 512.2 [M+H] + .
  • Step 8 Water (0.5 mL), Int 1a (239 mg, 0.78 mmol), Pd(dppf)Cl 2 (44 mg, 0.06 mmol) and potassium carbonate (181 mg, 1.31 mmol) were added to the reaction system of KH 1-8, and the mixture was stirred at 90°C for 16 h. After the reaction was completed, the reaction solution was concentrated and the crude product was purified by silica gel column chromatography to obtain KH 1-9. LCMS (ESI, m/z): 612.2 [M+H] + .
  • Step 9 Lithium hydroxide (20.16 mg, 0.48 mmol) was added to a mixed solution of KH 1-9 (150 mg, 0.24 mmol) in methanol (1.5 mL) and water (0.5 mL), and the mixture was stirred at 60°C for 1 h. After the reaction was completed, the solvent was evaporated under reduced pressure, and the crude product was purified by preparative HPLC to obtain KH 01.
  • Step 10 Compound KH01 (27 mg, 0.04 mmol) was chirally separated by SFC to obtain KH01-1 and KH01-2.
  • KH01-1 LCMS (ESI, m/z): 584.1 [M+H] + .
  • Example 2 2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2,5-difluorobenzyl)-1-((oxetan-2-yl)methyl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxylic acid (KH02), (R)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2,5-difluorobenzyl)-1-((oxetan-2-yl)methyl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxylic acid (KH02), Synthesis of carboxylic acid (KH02-1), (S)-2-(4-(6-((4-cyano-2-fluorobenzyl)oxy)pyridin-2-yl)-2,5-difluorobenzyl)
  • Step 1 Add sulfamic acid (3.2 g, 32.9 mmol) and sodium hypochlorite (4.9 g, 55.1 mmol) to a mixed solution of 3-fluoro-4-formylbenzoic acid methyl ester compound KH2-1 (5 g, 27.5 mmol) in ACN (50 mL)-H 2 O (10 mL), and stir the reaction at room temperature for 4 h. After the reaction is completed, adjust the pH to 3-4 with 1N hydrochloric acid solution. Add ethyl acetate (30 mL ⁇ 3) for extraction, combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain KH2-2.
  • LCMS ESI, m/z
  • Step 2 To a solution of KH 2-2 (4.9 g, 24.7 mmol) in DCM (50 mL) were added oxalyl chloride (62.8 g, 494.8 mmol) and DMF (20 mg, 0.27 mmol). The mixture was stirred at room temperature for 16 h. After the reaction was complete, the mixture was evaporated under reduced pressure and the crude product was used directly in the next step.
  • Step 3 Add intermediate KH 2-3 (4.1 g, 22.2 mmol) and triethylamine (4.5 g, 44.4 mmol) to a solution of tert-butyl 2-(4-bromo-2,5-difluorobenzyl)hydrazine-1-carboxylate (Int 2a, 7.5 g, 22.2 mmol) in DCM (50 mL). The reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain KH 2-4. LCMS (ESI, m/z): 460.7 [M+H-56] + .
  • Step 4 Add 4M hydrochloric acid in ethyl acetate (50 mL) to a solution of intermediate KH 2-4 (5 g, 9.6 mmol) in ethyl acetate (50 mL), and stir the mixture at room temperature for 2 h. After the reaction is complete, concentrate to obtain crude KH2-5.
  • LCMS (ESI, m/z): 417.1 [M+H] + .
  • Step 5 Dissolve the intermediate KH 2-5 (4.8 g, 11.51 mmol) in DMSO (50 mL), add DIEA (4.46 g, 34.53 mmol) to the reaction system, and stir the reaction at 150°C for 2 h. After the reaction is completed, add water (80 mL) to dilute the reaction system, extract with ethyl acetate (30 mL x 3), combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate the crude reaction solution to purify KH 2-6 by silica gel column chromatography. LCMS (ESI, m/z): 398.8 [M+H] + .
  • Step 6 Dissolve KH 2-6 (0.5 g, 1.26 mmol) in DMF (5 mL), add 2-iodomethyloxetane (0.32 g, 1.64 mmol) and anhydrous potassium carbonate (0.35 g, 2.52 mmol) to the reaction system at 50 °C. After stirring for 12 h, water (30 mL) was added to dilute the reaction system, and the mixture was extracted with ethyl acetate (20 mL ⁇ 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography to obtain KH 2-7.
  • Step 7 Dissolve the intermediate KH 2-7 (0.4 g, 0.86 mmol) in 1,4-dioxane (5 mL), add pinacol diboron (0.33 g, 1.28 mmol), KOAc (0.17 g, 1.71 mmol) and Pd(dppf)Cl 2 (0.06 g, 0.09 mmol) to the reaction system in sequence, and stir at 80°C for 2 h. After the reaction is complete, evaporate the solvent to obtain KH 2-8. LCMS (ESI, m/z): 515.3 [M+H] + .
  • Step 8 Dissolve the intermediate KH 2-8 (0.4 g, 0.77 mmol) in a mixed solvent of 1,4-dioxane (5 mL)-water (0.5 mL), and add Int 1a (0.24 g, 0.78 mmol), K 2 CO 3 (0.22 g, 1.55 mmol) and Pd(dppf)Cl 2 (0.06 g, 0.08 mmol) to the reaction system in sequence, and stir the reaction at 90°C for 2 h. After the reaction is completed, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography to obtain KH 2-9. LCMS (ESI, m/z): 615.0 [M+H] + .
  • Step 10 Dissolve the intermediate KH 2-9 (30 mg, 0.048 mmol) in a mixed solution of methanol (2.5 mL) and water (0.5 mL), add potassium carbonate (14 mg, 0.099 mmol) to the reaction system, and stir at 50 ° C for 3 h. After the reaction is completed, dilute the reaction system with water (10 mL), adjust the pH value, extract with ethyl acetate (10 mL x 3), wash the combined organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, and purify the concentrated crude product by preparative HPLC to obtain KH 2.
  • KH 2-9a 40 mg
  • KH 2-9b 70 mg
  • LCMS (ESI, m/z): 650.3[M+H] + .
  • Step 4 Add LiOH (7.9 mg, 0.33 mmol) in water (2 mL) to a solution of intermediate KH 3-2b (110 mg, 0.16 mmol) in THF (8 mL) and stir at room temperature for 1 h. After the reaction is complete, add 20 mL of water, adjust pH to 3 with 1 M hydrochloric acid, extract with ethyl acetate (30 mL x 3), wash the combined organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain KH 03-2.
  • Step 1 Dissolve the intermediate KH 2-6 (1 g, 2.51 mmol) in DMF (5 mL), add 2-(1-(iodomethyl)cyclopropyl)acetonitrile (0.41 g, 3.77 mmol) and anhydrous potassium carbonate (0.7 g, 5.06 mmol) to the reaction system, and stir the reaction at 50 ° C for 3 h. After the reaction is completed, dilute the reaction system with water (30 mL), extract with ethyl acetate (20 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product with a silica gel column to obtain KH 4-2.
  • LCMS (ESI, m/z): 490.2 [M+H] + .
  • Step 2 Dissolve the intermediate KH 4-2 (0.2 g, 0.40 mmol) in 1,4-dioxane (3 mL), add diboric acid pinacol ester (0.16 g, 0.61 mmol), KOAc (0.08 g, 0.81 mmol) and Pd(dppf)Cl 2 (0.03 g, 0.04 mmol) to the reaction system, and stir at 80°C for 2 h. After the reaction is completed, evaporate the reaction solution and use it directly in the next step.
  • LCMS (ESI, m/z): 538.2 [M+H] + .
  • Step 3 Dissolve the intermediate KH 4-3 (0.2 g, 0.37 mmol) in a mixed solvent of 1,4-dioxane (2 mL)-water (0.2 mL), and add Int 1a (0.12 g, 0.37 mmol), K 2 CO 3 (0.10 g, 0.74 mmol) and Pd(dppf) 2 Cl 2 (0.03 g, 0.04 mmol) to the reaction system in sequence, and stir at 90°C for 2 h. After the reaction is completed, evaporate the solvent under reduced pressure, and purify the crude product by silica gel column chromatography to obtain KH 4-4. LCMS (ESI, m/z): 638.1 [M+H] + .
  • Step 4 Dissolve the intermediate KH 4-4 (0.15 g, 0.23 mmol) in a methanol (2.5 mL)-water (0.5 mL) mixed solvent, then add K 2 CO 3 (0.06 mg, 0.47 mmol) into the reaction system and stir at room temperature for 3 h. After completion, the reaction system was diluted with water (10 mL), the pH was adjusted to 5-6 (1 M), extracted with ethyl acetate (10 mL x 3), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the reaction solution was concentrated. The crude product was purified by preparative HPLC to obtain KH04. LCMS (ESI, m/z): 624.2[M+H] + .
  • Step 1 Dissolve the intermediate KH 4-3 (0.1 g, 0.18 mmol) in a mixed solution of 1,4-dioxane (2 mL)-water (0.2 mL), and sequentially add 2-bromo-6-((4-chloro-2-fluorobenzyl)oxy)pyridine (0.06 g, 0.18 mmol), K 2 CO 3 (0.05 g, 0.36 mmol) and Pd(dppf)Cl 2 (0.014 g, 0.019 mmol) to the reaction system, and stir the reaction at 90°C for 2 h. After the reaction is complete, concentrate the reaction solution, and purify the crude product by silica gel column chromatography to obtain KH 5-2. LCMS (ESI, m/z): 647.2 [M+H] + .
  • Step 2 Dissolve the intermediate KH 5-2 (0.13 g, 0.2 mmol) in methanol (2.5 mL) and water (0.5 mL), add LiOH (0.02 mg, 0.47 mmol) to the reaction system, and stir at room temperature for 3 h. After the reaction is completed, dilute the reaction system with water (10 mL), adjust the pH to 5-6 with hydrochloric acid (1 M), extract with ethyl acetate (10 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, filter, and purify the concentrated filtrate by preparative HPLC to obtain KH05.
  • LCMS (ESI, m/z): 633.02[M+H] + .
  • Step 1 Dissolve the intermediate Int 2c (3.41 g, 10.24 mmol) and triethylamine (2.59 g, 25.70 mmol) in dichloromethane (40 mL), add the dichloromethane solution of the intermediate KH 2-3 (2.79 g, 12.85 mmol) dropwise at 0°C, and keep warm for 2 h. After the reaction is completed, add water (30 mL) to quench, extract with dichloromethane (3 ⁇ 150 mL), wash the organic phase with brine, dry with sodium sulfate, filter, concentrate to remove the solvent, and purify the residue by silica gel column chromatography to obtain KH 6-2.
  • Step 2 Add intermediate KH 6-2 (4.6 g, 8.98 mmol) to a solution of hydrogen chloride in ethyl acetate (2N, 50 mL) and stir at room temperature for 12 h. After the reaction is complete, concentrate under reduced pressure to obtain crude KH 6-3.
  • LCMS (ESI, m/z): 413.2 [M+H] + .
  • Step 3 Add DIEA (4.77 g, 34.57 mmol) to a DMSO (30 mL) solution of the intermediate KH 6-3 (2.8 g, 6.80 mmol) and react at 150°C for 2 h. After the reaction is complete, add water (200 mL), extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify the crude product by silica gel column chromatography to obtain KH 6-4.
  • LCMS (ESI, m/z): 393.2 [M+H] + .
  • Step 4 Add potassium carbonate (289 mg, 2.04 mmol) and 2-(1-(iodomethyl)cyclopropyl)acetonitrile (338 mg, 1.53 mmol) to a DMF (5 mL) solution of the intermediate KH 6-4 (400 mg, 1.02 mmol), and react at room temperature for 4 h. Add water (200 mL), extract with ethyl acetate, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and purify the crude product by silica gel column chromatography to obtain KH 6-5P1 and KH 6-5P2.
  • Step 5 Add potassium acetate (81 mg, 0.83 mmol), diboronic acid pinacol (136 mg, 0.54 mmol) and Pd(dppf)Cl 2 (30 mg, 0.04 mmol) to a solution of KH 6-5P1 (200 mg, 0.41 mmol) in dioxane (3 mL), and stir at 80°C for 4 h under nitrogen atmosphere to generate KH 6-6P1.
  • KH 6-6P2 was obtained by the same method. Both KH 6-6P1 and KH 6-6P2 were used directly in the next step without further treatment.
  • Step 6 Add 1,4-dioxane (3 mL)-water (0.3 mL) mixed solution to the KH 6-6P1 system, then add compound Int 1a (164 mg, 0.54 mmol), potassium carbonate (114 mg, 0.83 mmol), Pd(dppf)Cl 2 (30 mg, 0.04 mmol), and stir at 90°C for 4 h under nitrogen protection. After the reaction is completed, concentrate and purify by silica gel column chromatography to obtain KH 6-7P1. LCMS (ESI, m/z): 633.6 [M+H] + .
  • Step 7 Add potassium carbonate (78 mg, 0.57 mmol) and water (2 mL) to a methanol solution (2 mL) of KH 6-7P1 (180 mg, 0.31 mmol) and stir at 50°C for 2 h. After the reaction is complete, adjust the pH to 4 with 1N hydrochloric acid. Then 20 mL of water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC to obtain KH06-1. LCMS (ESI, m/z): 619.6[M+H] + .
  • KH06-2 was obtained by the same method using KH 6-7P2 as the starting material.
  • Step 1 Add triethylamine (2.8 g, 27.7 mmol) and raw material KH 2-3 (3 g, 13.8 mmol) to a DCM (30 mL) solution of intermediate Int 2c (2.6 g, 8.2 mmol), and stir the reaction at room temperature for 2 h. After the reaction is complete, concentrate the reaction solution under reduced pressure, and purify the crude product by silica gel column chromatography to obtain KH 7-2.
  • Step 2 3M hydrochloric acid ethyl acetate (10 mL) was added to a solution of the intermediate KH 7-2 (1.8 g, 3.6 mmol) in ethyl acetate (10 mL), and the mixture was stirred at room temperature for 12 h. After the reaction was completed, the mixture was concentrated to obtain crude KH 7-3.
  • LCMS (ESI, m/z): 399.0 [M+H] + .
  • Step 3 The intermediate KH 7-3 (0.4 g, 1.0 mmol) was dissolved in DMSO (10 mL), DIEA (0.38 g, 3.0 mmol) was added to the reaction system, and the reaction was stirred at 150 ° C for 2 h. After the reaction was completed, water (30 mL) was added to dilute the reaction system, and ethyl acetate (20 mL ⁇ 3) was used for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography to obtain KH 7-4. LCMS (ESI, m/z): 379.0[M+H] + .
  • Step 4 Dissolve the intermediate KH 7-4 (0.1 g, 0.26 mmol) in DMF (5 mL), add 2-(iodomethyl)butylene oxide (0.67 g, 0.33 mmol) and potassium carbonate (0.73 g, 0.52 mmol) to the reaction system, and stir the reaction at 50°C for 12 h. After the reaction is complete, add water (30 mL) to dilute the reaction system, extract with ethyl acetate (20 mL ⁇ 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product by silica gel column chromatography to obtain KH 7-5.
  • LCMS (ESI, m/z): 449.2 [M+H] + .
  • Step 5 Dissolve the intermediate KH 7-5 (0.05 g, 0.11 mmol) in 1,4-dioxane (2 mL), and sequentially add pinacol diboron (0.04 g, 0.16 mmol), KOAc (0.02 g, 0.22 mmol) and Pd(dppf)Cl 2 (0.008 g, 0.01 mmol) was added to the reaction system and the reaction was stirred at 80°C for 2 h. After the reaction was completed, the product was concentrated to obtain KH 7-6. LCMS (ESI, m/z): 497.3 [M+H] + .
  • Step 6 Dissolve the intermediate KH 7-6 (0.05 g, 0.1 mmol) in 1,4-dioxane (1 mL) and water (0.1 mL), and add the intermediate Int 1b (0.03 g, 0.1 mmol), K 2 CO 3 (0.02 g, 0.15 mmol) and Pd(dppf)Cl 2 (6 mg, 0.008 mmol) to the reaction system in sequence, and stir the reaction at 90°C for 2 h. After the reaction is completed, concentrate, and purify the crude product by silica gel column to obtain KH 7-7.
  • LCMS (ESI, m/z): 658.3 [M+H] + .
  • Step 7 Dissolve the intermediate KH 7-7 (0.05 g, 0.07 mmol) in a mixed solution of methanol (2.5 mL) and water (0.5 mL), add LiOH (3.6 mg, 0.15 mmol) to the reaction system, and stir at room temperature for 3 h. After the reaction is completed, dilute with water (10 mL), adjust the pH to 3-4 with 1 M hydrochloric acid, extract with ethyl acetate (10 mL ⁇ 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product by preparative HPLC to obtain KH07.
  • LCMS ESI, m/z
  • Step 1 Add 2-(1-(iodomethyl)cyclopropyl)acetonitrile (166.68 mg, 0.84 mmol) and potassium carbonate (209.76 mg, 1.52 mmol) to a DMF (5 mL) solution of the intermediate KH 6-4 (300 mg, 0.76 mmol), and react the mixture at 50° C. for 2 h. After the reaction is complete, add water, extract with ethyl acetate, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The crude product is purified by silica gel column chromatography to obtain KH 8-2. LCMS (ESI, m/z): 463.2 [M+H] + .
  • Step 2 Add potassium acetate (85 mg, 0.87 mmol), biboronic acid pinacol ester (143 mg, 0.56 mmol) and Pd(dppf)Cl 2 (32 mg, 0.04 mmol) to a solution of intermediate KH 8-2 (200 mg, 0.43 mmol) in dioxane (2 mL), and stir at 80°C for 4 h under nitrogen protection to generate KH 8-3. Used directly in the next step without further treatment. LCMS (ESI, m/z): 510.3 [M+H] + .
  • Step 3 Add 1,4-dioxane (1 mL) and water (0.1 mL) to the intermediate KH 8-3, then add compound Int 1a (156 mg, 0.51 mmol), potassium carbonate (108 mg, 0.78 mmol), Pd(dppf)Cl 2 (29 mg, 0.04 mmol), and stir at 90°C for 4 h under nitrogen atmosphere. After the reaction is completed, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography to obtain KH 8-4. LCMS (ESI, m/z): 610.6 [M+H] + .
  • Step 4 Add potassium carbonate (67 mg, 0.49 mmol) and water (1 mL) to a methanol solution (1 mL) of the intermediate KH 8-4 (148 mg, 0.24 mmol) and stir at 50°C for 2 h. After the reaction, the reaction solution was diluted with water, the pH value was adjusted to 5-6, and extracted with ethyl acetate. The organic phase was dried and filtered, the filtrate was concentrated, and the crude product was purified by preparative HPLC. Got KH08. LCMS (ESI, m/z): 596.5[M+H] + .
  • Step 1 Add intermediate Int 1c (122 mg, 0.33 mmol), potassium carbonate (71 mg, 0.51 mmol), and Pd(dppf)Cl 2 (19 mg, 0.03 mmol) to a mixed solution of intermediate KH 8-3 (130 mg, 0.25 mmol) in 1,4-dioxane (2 mL)-water (0.2 mL), replace with N 2 three times, and stir at 90°C for 4 h. After the reaction is completed, the crude product after reduced pressure concentration is purified by silica gel column chromatography to obtain KH 9-2. LCMS (ESI, m/z): 671.6 [M+H] + .
  • Step 2 Add potassium carbonate (66 mg, 0.48 mmol) and water (2 mL) to a methanol solution (2 mL) of the intermediate KH 9-2 (130 mg, 0.19 mmol), and stir at 50°C for 2 h. After the reaction, the reaction solution was diluted with water, the pH value was adjusted to 5-6, and extracted with ethyl acetate. The organic phase was dried and filtered, the filtrate was concentrated, and the crude product was purified by preparative HPLC to obtain KH09. LCMS (ESI, m/z): 657.6 [M+H] + .
  • LCMS ESI, m/z
  • Step 1 Dissolve the intermediate KH 7-6 (0.1 g, 0.2 mmol) in a mixed solution of 1,4-dioxane (2 mL) and water (0.2 mL), and add the intermediate Int 1a (0.12 g, 0.4 mmol), K 2 CO 3 (0.05 g, 0.4 mmol) and Pd(dppf)Cl 2 (0.015 g, 0.02 mmol) to the reaction system in sequence, and stir the reaction at 90°C for 2 h. After the reaction is complete, concentrate the reaction solution, and purify the crude product by silica gel column chromatography to obtain KH 11-2. LCMS (ESI, m/z): 597.1 [M+H] + .
  • Step 2 Dissolve the intermediate KH 11-2 (0.1 g, 0.17 mmol) in acetonitrile (2 mL), add TBD (0.9 M, 0.4 mL) to the reaction system, and stir at room temperature for 2 h. After the reaction is completed, dilute with water (5 mL), adjust the pH to 3 with 1N hydrochloric acid, extract with ethyl acetate (5 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify the crude product by preparative HPLC to obtain KH11.
  • LCMS ESI, m/z: 583.2[M+H] + .
  • Step 1 Dissolve the intermediate KH 7-6 (0.1 g, 0.2 mmol) in a mixed solution of 1,4-dioxane (2 mL) and water (0.2 mL), and sequentially add the intermediate Int 1d (0.065 g, 0.2 mmol), K 2 CO 3 (0.05 g, 0.4 mmol) and Pd(dppf)Cl 2 (0.015 g, 0.02 mmol) to the reaction system, and stir the reaction at 90°C for 2 h. After the reaction is complete, concentrate the reaction solution, and purify the crude product by silica gel column chromatography to obtain KH 12-2. LCMS (ESI, m/z): 615.1 [M+H] + .
  • Step 2 Dissolve the intermediate KH 12-2 (0.1 g, 0.16 mmol) in acetonitrile (2 mL), add TBD (0.9 M, 0.4 mL) to the reaction system, and stir at room temperature for 2 h. After the reaction is completed, dilute with water (5 mL), adjust the pH to 3 with 1N hydrochloric acid, extract with ethyl acetate (5 mL ⁇ 3), wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify the crude product by preparative HPLC to obtain KH12.
  • LCMS ESI, m/z
  • Step 2 To a solution of intermediate KH 13-2 (310 mg, 0.50 mmol) in acetonitrile (3 mL), add an aqueous solution of TBD (0.9 M, 1.2 mL), and stir the mixture at room temperature for 2 h. Concentrate to obtain a crude product, which is purified by Prep-HPLC to obtain KH13.
  • LCMS (ESI, m/z): 613.1 [M+H] + .
  • Step 1 Add K 2 CO 3 (670 mg, 4.855 mmol) to a DMF (10 mL) solution of KH 7-4 (920 mg, 2.427 mmol) and intermediate Int 1f (1.191 g, 3.641 mmol), and stir at 50°C for 2 h. After the reaction is complete, add water (30 mL), extract with ethyl acetate (30 mL ⁇ 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product by silica gel column to obtain KH 14-2.
  • LCMS (ESI, m/z): 577.1[M+H] + .
  • Step 2 Add TBAF (1.35 mL, 1 M) in THF to a solution of intermediate KH 14-2 (520 mg, 0.901 mmol) in THF (10 mL) and stir at room temperature for 2 h. After the reaction is complete, add water (30 mL), extract with ethyl acetate (30 mL ⁇ 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product with a silica gel column to obtain KH 14-3.
  • LCMS (ESI, m/z): 463.1 [M+H] + .
  • Step 3 Add DAST (146 mg, 0.907 mmol) to a solution of intermediate KH 14-3 (210 mg, 0.454 mmol) in dichloromethane (8 mL), and stir at -78 °C for 1 h. After the reaction is complete, quench the reaction with saturated aqueous NaHCO 3 solution, extract with dichloromethane (30 mL ⁇ 3), dry with sodium sulfate, concentrate, and purify the crude product by silica gel column to obtain KH14-4.
  • LCMS (ESI, m/z): 465.1[M+H] + .
  • Step 4 Pd(dppf)Cl 2 (28 mg, 0.0387 mmol) and KOAc (51 mg, 0.516 mmol) were added to a solution of intermediate KH 14-4 (120 mg, 0.258 mmol) and biboronic acid pinacol ester (98 mg, 0.387 mmol) in dioxane (10 mL), and stirred at 80°C for 6 h under nitrogen atmosphere. After the reaction was completed, the obtained KH 14-5 was directly used in the next step.
  • Step 5 Add Int 1a (71 mg, 0.232 mmol), Pd(dppf)Cl 2 (28 mg, 0.0387 mmol), potassium carbonate (71 mg, 0.516 mmol), water (1 mL) to the reaction solution containing the intermediate KH 14-5 (11.5 mL), and stir at 90°C for 12 h under nitrogen atmosphere. After the reaction is completed, add water (30 mL), extract with ethyl acetate (40 mL ⁇ 3), dry with sodium sulfate, concentrate, and purify the crude product by silica gel column to obtain KH 14-6.
  • Step 6 Add TBD (0.45 mL, 0.9 M in H 2 O) to a solution of the intermediate KH 14-6 (110 mg, 0.179 mmol) in acetonitrile (8 mL) and stir at room temperature for 2 h. After the reaction is complete, add water (20 mL), adjust the pH to 3 with HCl (1 M), extract with ethyl acetate (30 mL ⁇ 3), wash with saturated brine, dry with sodium sulfate, concentrate, and purify the crude product by Prep-HPLC to obtain KH14.
  • LCMS (ESI, m/z): 599.2[M+H] + .
  • Step 2 The intermediate KH 15-2a was separated by SFC to obtain KH 15-3a (retention time 1.288 min) and KH15-3b (retention time 2.09 min).
  • LCMS (ESI, m/z): 610.1 [M+H] + .
  • Step 1 Add a DCM (20 mL) solution of the intermediate KH 2-3 to a DCM (20 mL) solution of the intermediate Int 2e (1.45 g, 4.545 mmol) and TEA (1.02 g, 10.10 mmol) at 0°C, and stir at room temperature for 2 h. After the reaction is complete, add water (30 mL), extract with DCM (40 mL ⁇ 3), wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify on a silica gel column to obtain KH 16-2.
  • LCMS (ESI, m/z): 499.1[M+H] + .
  • Step 2 Add ethyl acetate (25 mL, 3 M) solution of hydrochloric acid to a solution of intermediate KH 16-2 (2.0 g, 4.008 mmol) in methanol (10 mL), and stir the mixture at room temperature for 2 h. After the reaction is complete, concentrate, add water (30 mL), adjust pH to 8 with saturated NaHCO 3 aqueous solution, extract with DCM (50 mL ⁇ 3), wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain KH 16-3.
  • LCMS (ESI, m/z): 399.0[M+H] + .
  • Step 3 Add DIEA (1.0 g, 7.769 mmol) to a DMSO (15 mL) solution of the intermediate KH 16-3 (1.5 g, 3.759 mmol) and stir at 150°C for 3 h. After the reaction is complete, add water (40 mL), extract with EA (50 mL ⁇ 3), wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify the crude product by normal phase column to obtain KH 16-4.
  • LCMS (ESI, m/z): 379.0 [M+H] + .
  • Step 4 Add 2-iodomethaneoxetane (367 mg, 1.852 mmol) to a DMF (10 mL) solution of the intermediate KH 16-4 (520 mg, 1.425 mmol) and K 2 CO 3 (393 mg, 2.850 mmol), and stir at 50°C for 16 h. After the reaction is complete, add water (30 mL), extract with EA (40 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product with a normal phase column to obtain KH 16-5.
  • LCMS (ESI, m/z): 449.0 [M+H] + .
  • Step 5 Pd(dppf)Cl 2 (78 mg, 0.107 mmol) and KOAc (140 mg, 1.425 mmol) were added to a solution of intermediate KH 16-5 (320 mg, 0.713 mmol) and biboronic acid pinacol ester (272 mg, 1.069 mmol) in 1,4-dioxane (15 mL), and stirred at 80°C for 12 h under nitrogen atmosphere. After the reaction was completed, the KH 16-6 reaction solution was directly used in the next step.
  • Step 6 Add intermediate Int 1a (186 mg, 0.606 mmol), Pd(dppf)Cl 2 (78 mg, 0.107 mmol), K 2 CO 3 (197 mg, 1.425 mmol) and H 2 O (1.5 mL) to the reaction solution of intermediate KH 16-6 (15 mL, 0.713 mmol), and stir at 90° C. for 2 h under nitrogen atmosphere. After the reaction is completed, concentrate, and purify the crude product by silica gel column chromatography to obtain KH 16-7. LCMS(ESI,m/z):597.2[M+H] + .
  • Step 7 Add LiOH (21 mg, 0.871 mmol) in H 2 O (2 mL) to a solution of intermediate KH 16-7 (260 mg, 0.436 mmol) in THF (10 mL), and stir at room temperature for 1 h. After the reaction is complete, add water (20 mL), adjust pH to 3 with 1 M hydrochloric acid, extract with EA (40 mL ⁇ 3), wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify by preparative HPLC to obtain KH16. LCMS(ESI,m/z):583.2[M+H] + .
  • Step 1 Add 2 mL of potassium tert-butoxide (730 mg, 6.53 mmol) in tetrahydrofuran to a solution of raw material KH 17-1 (1 g, 5.43 mmol) and 2-fluoro-6-bromopyridine (956.5 mg, 5.43 mmol) in dioxane (10 mL), stir at room temperature, and gradually heat to 40 ° C for 0.5 h. After the reaction is completed, water (10 mL) and ethyl acetate (10 mL x 3) are added for extraction, and the organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column to obtain KH 17-2.
  • Step 2 Add lithium hydroxide (125 mg, 5.35 mmol) dissolved in water (2 mL) to a solution of intermediate KH 17-2 (890 mg, 2.62 mmol) in methanol (10 mL), and stir at room temperature for 2 h. After the reaction is completed, adjust pH to 3 with 1N hydrochloric acid, add water (10 mL) and ethyl acetate (10 mL x 3) for extraction, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain KH 17-3.
  • LCMS (ESI, m/z): 341.9[M+H] + .
  • Step 3 TCFH (840 mg, 3.0 mmol) and NMI (566 mg, 6.9 mmol) were added to a solution of intermediate KH 17-3 (750 mg, 2.3 mmol) and methoxymethylamine hydrochloride (337 mg, 3.45 mmol) in dichloromethane (10 mL), and the mixture was stirred at room temperature for 1 h. After the reaction was completed, water (10 mL) and ethyl acetate (10 mL x 3) were added for extraction, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and purified by silica gel column chromatography to obtain KH 17-4.
  • Step 4 Slowly add cyclopropylmagnesium bromide (1.54 g, 10.6 mmol) to a solution of intermediate KH 17-4 (780 mg, 2.12 mmol) in ultra-dry tetrahydrofuran (10 mL), and stir at 0°C for 3 h. Then, add saturated aqueous ammonium chloride solution (10 mL), extract with ethyl acetate (10 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify the crude product by silica gel column chromatography to obtain KH 17-5.
  • LCMS (ESI, m/z): 351.9 [M+H] + .
  • Step 5 Add intermediate KH 2-8 (200 g, 0.43 mmol), potassium carbonate (118.7 mg, 0.56 mmol) and Pd(dppf)Cl 2 (31.4 mg, 0.043 mmol) to a mixed solution of intermediate KH 17-5 (150 mg, 0.43 mmol) in dioxane (3 mL) and water (0.3 mL), and stir at 90°C for 2.5 h. After the reaction is completed, the reaction solution is concentrated and the crude product is purified by silica gel column chromatography to obtain KH 17-6. LCMS (ESI, m/z): 658.2 [M+H] + .
  • Step 6 Add an aqueous solution of TBD (0.9 mol, 0.8 mL) to a solution of the intermediate KH 17-6 (220 mg, 0.334 mmol) in acetonitrile (2 mL) and stir at room temperature for 2.5 h. After the reaction is complete, add water (5 mL), extract with ethyl acetate (5 mL ⁇ 3), wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate. The crude product obtained by concentration is purified by preparative HPLC to obtain KH17. LCMS (ESI, m/z): 644.1 [M+H] + .
  • Step 1 Pd(dppf)Cl 2 (2.34 g, 4.23 mmol) was added to a methanol (100 mL) solution of the raw material KH 18-1 (10 g, 42.3 mmol) and TEA (21.398 g, 211.9 mmol), and stirred at 120°C for 12 h under a carbon monoxide pressure atmosphere. After the reaction was completed, the crude product was filtered and concentrated to obtain KH 18-2, which was purified by Prep-HPLC. LCMS (ESI, m/z): 214.8 [MH] - .H NMR (400 MHz, DMSO) ⁇ 7.77-7.60 (m, 2H), 3.90 (s, 3H).
  • Step 2 Add oxalyl chloride (94.1 g, 740.7 mmol) and catalytic amount of DMF to a solution of intermediate KH 18-2 (8 g, 37.0 mmol) in dichloromethane (80 mL), and stir at room temperature for 12 h. After the reaction is complete, concentrate to obtain crude KH 18-3, which is directly used for the next step.
  • Step 3 Add intermediate KH 18-3 dropwise to a solution of intermediate Int 2b (8.7 g, 27.4 mmol) and TEA (5.52 g, 54.7 mmol) in dichloromethane (60 mL), and stir at room temperature for 2.5 h. After the reaction is complete, add water (100 mL), extract with dichloromethane (100 mL x 3), wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and purify the crude product by silica gel column chromatography to obtain KH 18-4.
  • LCMS (ESI, m/z): 580.0 [M+Na+ACN] + .
  • Step 4 Add hydrogen chloride ethyl acetate solution (100 mL) to a solution of intermediate KH 18-4 (13.7 g, 26.5 mmol) in ethyl acetate (100 mL) and stir at room temperature for 12 h. After the reaction is complete, filter to obtain KH 18-5.
  • LCMS (ESI, m/z): 419.0 [M+H] + .
  • Step 5 Add DIEA (928 mg, 7.2 mmol) to a solution of intermediate KH 18-5 (1 g, 2.4 mmol) in dimethyl sulfoxide (10 mL), and stir at 150°C for 2 h. After the reaction is complete, add water (20 mL), extract with ethyl acetate (20 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain a crude product, which is purified by preparative HPLC to obtain KH 18-6.
  • LCMS (ESI, m/z): 397.0[M+H] + .
  • Step 6 Add DIEA (304.6 mg, 2.36 mmol) to a solution of intermediate KH 18-6 (470 mg, 1.18 mmol) and 2-iodomethyloxetane (257.2 mg, 1.3 mmol) in dimethyl sulfoxide (5 mL), and stir at 50°C for 12 h. After the reaction is complete, extract with water (10 mL) and ethyl acetate (10 mL ⁇ 3), wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and purify the crude product by silica gel column chromatography to obtain KH 18-7.
  • LCMS (ESI, m/z): 469.0 [M+H] + .
  • Step 7 Add potassium acetate (66.3 mg, 0.68 mmol) and Pd(dppf)Cl 2 (24.9 mg, 0.034 mmol) to a solution of intermediate KH 18-7 (158 mg, 0.34 mmol) and Int 1a (129 mg, 0.51 mmol) in tetrahydrofuran (3 mL), and stir at 80°C for 2.5 h. After the reaction is complete, the crude KH 18-8 is used directly in the next step. LCMS (ESI, m/z): 515.2 [M+H] + .
  • Step 8 Add water (0.03 mL), potassium carbonate (93.4 mg, 0.68 mmol) and Pd(dppf)Cl 2 (24.9 mg, 0.034 mmol) to the reaction solution of the previous step, and stir at 90°C for 2.5 h. After the reaction is completed, add water (10 mL) and ethyl acetate (10 mL ⁇ 3) for extraction, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and purify the crude product by silica gel column chromatography to obtain KH 18-9.
  • LCMS (ESI, m/z): 615.1 [M+H] + .
  • Step 9 Add an aqueous solution of TBD (0.9 mol, 0.4 ml) to a solution of the intermediate KH 18-9 (120 mg, 0.195 mmol) in acetonitrile (2 mL) and stir at room temperature for 2.5 h. After the reaction is complete, add water (5 mL) and ethyl acetate (5 mL ⁇ 3) for extraction, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, filter, and concentrate. The crude product is purified by preparative HPLC to obtain KH18. LCMS (ESI, m/z): 601.2[M+H] + .
  • Step 1 Add intermediate Int 2d (1.56 mg, 4.62 mmol) and triethylamine (1.2 mL) to a solution of intermediate KH 2-3 (1 g, 4.62 mmol) in dichloromethane (10 mL) and stir at room temperature for 2 h. After the reaction is complete, vacuum concentrate to obtain a crude product which is purified by silica gel column chromatography to obtain KH 19-2. LCMS (ESI, m/z): 416.9 [M+1] + .
  • Step 2 Add ethyl acetate (20 mL) of hydrogen chloride to a solution of intermediate KH 19-2 (2 g, 3.87 mmol) in ethyl acetate (20 mL) and react at room temperature for 2 h. After the reaction is complete, filter the reaction solution, rinse the filter cake with ethyl acetate, and dry the filter cake to obtain KH 19-3.
  • LCMS (ESI, m/z): 419.0 [M+1] + .
  • Step 3 Add DIEA (1.2 mL) to a solution of intermediate KH 19-3 (950 mg, 2.28 mmol) in dimethyl sulfoxide (10 mL) and react at 150°C for 2 h. After the reaction is complete, pour the reaction solution into water, add ethyl acetate for extraction, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain a crude product; the crude product is then purified by silica gel column chromatography to obtain KH 19-4.
  • LCMS (ESI, m/z): 397.0 [M+1] + .
  • Step 4 To a solution of KH 19-4 (100 mg, 0.25 mmol) in DMF (2 mL) was added 2-iodomethyl Oxetane (65 mg, 0.33 mmol) and potassium carbonate (70 mg, 0.50 mmol) were reacted at 50°C for 4 h. After the reaction was complete, the reaction solution was poured into water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography to obtain KH 19-5. LCMS (ESI, m/z): 469.0 [M+1] + .
  • Step 5 Add pinacol diboronate (90 mg, 0.35 mmol), potassium acetate (46 mg, 0.47 mmol) and Pd(dppf)Cl 2 (17 mg, 0.024 mmol) to a solution of intermediate KH 19-5 (110 mg, 0.24 mmol) in 1,4-dioxane (3 mL), and react at 80° C. under nitrogen protection for 2 h. After the reaction is complete, the product KH 19-6 is directly used for the next step.
  • LCMS (ESI, m/z): 433.0 [M+1] + .
  • Step 6 Add intermediate Int 1a (78 mg, 0.25 mmol), potassium carbonate (70 mg, 0.51 mmol) and Pd(dppf)Cl 2 (18 mg, 0.025 mmol) to a mixed solution of intermediate KH19-6 (110 mg, 0.25 mmol) in 1,4-dioxane (2 mL) and water (0.5 mL), and react at 90° C. under nitrogen protection for 12 h. After the reaction is complete, concentrate and purify the crude product by silica gel column chromatography to obtain KH 19-7. LCMS (ESI, m/z): 615.2 [M+1] + .
  • LCMS (ESI, m/z): 601.1[M+1] + .
  • Step 1 Add t-BuOK (940 mg, 8.4 mmol) to a solution of raw material KH 20-1 (1 g, 7.0 mmol) and 2-fluoro-6-bromopyridine (1.2 g, 7.0 mmol) in dioxane (10 mL), and stir the mixture at 40°C for 0.5 h. After the reaction is complete, add water (10 mL), extract with ethyl acetate (10 mL x 3), wash the combined organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify the crude product by silica gel column chromatography to obtain KH20-2.
  • Step 2 Add water (0.02 mL), potassium carbonate (93.4 mg, 0.68 mmol) and Pd(dppf)Cl 2 (24.9 mg, 0.034 mmol) to a solution of intermediate KH 20-2 (45 mg, 0.15 mmol) and intermediate KH 2-8 (77 mg, 0.15 mmol) in dioxane (2 ml), and stir at 90°C for 2.5 h. After the reaction is complete, add water (5 mL), extract with ethyl acetate (5 mL x 3), wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify the crude product by silica gel column chromatography to obtain KH 20-3.
  • LCMS (ESI, m/z): 607.1 [M+H] + .
  • Step 1 Dissolve the raw material KH 21-1 (2g, 7.19mmol) in DMF (20mL), add cuprous cyanide (1.5g, 16.85mmol) to the reaction system, and stir the reaction at 140°C for 2h. After the reaction is completed, cool to room temperature and add water (80mL) to quench, extract with ethyl acetate (30mL ⁇ 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product with a silica gel column to obtain KH 21-2.
  • Step 2 Dissolve the intermediate KH 21-2 (1.2 g, 6.77 mmol) in acetonitrile (10 mL), add deuterated iodomethane (1.9 g, 13.10 mmol) and cesium carbonate (4.4 g, 13.50 mmol) into the reaction system, and stir at room temperature for 6 h. After the reaction is complete, concentrate the product, and purify the crude product by silica gel column to obtain KH 21-3.
  • Step 3 Dissolve the intermediate KH 21-3 (1 g, 5.15 mmol) in THF (10 mL), add LiBH 4 (5 mL, 2.0 M) and MeOH (0.33 g, 10.31 mmol) to the reaction system, and stir at 50 °C for 2 h. After the reaction is completed, cool to room temperature, add saturated ammonium chloride solution (50 mL) to quench, extract with ethyl acetate (30 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify the crude product with a silica gel column to obtain KH 21-4.
  • Step 4 Dissolve the intermediate KH 21-4 (0.2 g, 1.19 mmol) in 1,4-dioxane (3 mL), add 2-bromo-6-fluoropyridine (0.21 g, 1.19 mmol) and potassium tert-butoxide (1.4 mL, 1 M) into the reaction system, and react at 45 °C. After the reaction was completed, the mixture was cooled to room temperature and concentrated. The crude product was purified by silica gel column to obtain KH 21-5. LCMS (ESI, m/z): 322.0 [M+H].
  • Step 5 Dissolve the intermediate KH 21-5 (0.22 g, 0.67 mmol) in 1,4-dioxane (3 mL) and water (0.3 mL), add the intermediate KH 2-8 (0.3 g, 0.64 mmol), potassium carbonate (0.17 g, 1.28 mmol) and Pd(dppf)Cl 2 (0.05 g, 0.06 mmol) to the reaction system, and stir at 90°C for 2 h. After the reaction is complete, concentrate, and purify the crude product by silica gel column to obtain KH 21-6. LCMS (ESI, m/z): 630.2 [M+H].
  • Step 6 The intermediate KH 21-6 (0.2 g) was separated by SFC [model: C2AS, conditions: 1525% MeOH DEA C2AS] to obtain KH 21-6a and KH 21-6b with retention times of 1.597 min and 2.553 min, respectively.
  • Step 7 Dissolve the intermediate KH 21-6b (0.1 g, 0.15 mmol) in acetonitrile (2 mL), add TBD (0.9 M, 0.4 mL) to the reaction system, and stir at room temperature for 2 h. After the reaction is completed, dilute with water (5 mL), adjust the pH to 5-6 with dilute acetic acid, extract with ethyl acetate (5 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product by Prep-HPLC to obtain KH21-2.
  • LCMS (ESI, m/z): 616.2 [M+H] + .
  • Example 3 The same synthesis method as in Example 3 was adopted, and the intermediates Int 1f-1 to Int 1f-4 were used as raw materials to react with the intermediate KH 2-8 (or the S-configured intermediate KH 2-8b) to obtain the target molecules KH23 to KH27, respectively.
  • Example 28 Preparation of (S)-2-(4-(6-((4-cyano-furo[3,2-c]pyridin-7-yl)methoxy)pyridin-2-yl)-2,5-difluorobenzyl)-1-((oxetane-2-yl)methyl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxylic acid (KH28)
  • Step 1 Dissolve the raw material KH 28-1 (10 g, 73.74 mmol) in acetonitrile (100 mL), add NBS (14.6 g, 73.74 mmol) dropwise at 0°C and stir for 1 h. After the reaction is complete, concentrate and slurry the crude product with methanol to obtain KH 28-2.
  • LCMS (ESI, m/z): 213.9 [M+H] + .
  • Step 2 Dissolve the intermediate KH 28-2 (3 g, 14.01 mmol) in ethanol (30 mL), add triethylamine (7 g, 69.17 mmol) and Pd(dppf)Cl 2 (1 g, 1.36 mmol) into the reaction system, and stir at 100 °C for 16 h in a carbon monoxide environment. After the reaction is complete, filter and concentrate, and the crude product is slurried with methyl tert-butyl ether to obtain KH 28-3.
  • LCMS (ESI, m/z): 208.1 [M+H].
  • Step 3 KH 28-3 (2 g, 9.61 mmol) was dissolved in phosphorus oxychloride (5 mL), and the reaction was stirred under reflux at 120 ° C for 1 h. After the reaction was completed, the mixture was cooled to room temperature and concentrated. The crude product was slowly added to an ice-water bath (80 mL) to quench, extracted with ethyl acetate (30 mL ⁇ 3), washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column to obtain KH 28-4.
  • LCMS (ESI, m/z): 226.0 [M+H].
  • Step 4 Dissolve the intermediate KH 28-4 (0.5 g, 2.21 mmol) in THF (10 mL), add LiBH 4 (2.6 mL, 2.0 M) and MeOH (0.17 g, 5.31 mmol) to the reaction system, and stir at 50 °C for 1 h. After the reaction is completed, cool to room temperature, add saturated ammonium chloride solution (30 mL) to quench, extract with ethyl acetate (20 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify the crude product by silica gel column to obtain KH 28-5.
  • LCMS (ESI, m/z): 184.8 [M+H].
  • Step 5 Dissolve the intermediate KH 28-5 (0.23 g, 1.25 mmol) in DMF (3 mL), add zinc cyanide (0.29 g, 2.50 mmol) and Pd(dppf)Cl 2 (0.09 mg, 0.12 mmol) to the reaction system, and stir the reaction at 120°C for 3 h. After the reaction is completed, cool to room temperature, add water (20 mL) to dilute, extract with ethyl acetate (10 mL ⁇ 3), wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify the crude product by silica gel column to obtain KH 28-6.
  • LCMS (ESI, m/z): 175.1 [M+H].
  • Step 6 Dissolve the intermediate KH 28-6 (0.18 g, 1.02 mmol) in 1,4-dioxane (2 mL), add 2-fluoro-6-bromopyridine (0.18 g, 1.02 mmol) and potassium tert-butoxide (1 mL, 1 M) to the reaction system, and stir the reaction at 45°C for 30 min. After the reaction is completed, cool to room temperature, concentrate, and the crude product is purified by silica gel column to obtain KH 28-7. LCMS (ESI, m/z): 330.0 [M+H].
  • Step 7 Dissolve the intermediate KH 28-7 (0.13 g, 0.27 mmol) in 1,4-dioxane (2 mL) and water (0.2 mL), add the intermediate KH 2-8b (0.1 g, 0.27 mmol), potassium carbonate (0.07 g, 0.55 mmol) and Pd(dppf)Cl 2 (0.02 g, 0.02 mmol) into the reaction system, and stir at 90°C for 2 h. After the reaction is complete, concentrate. The crude product was purified by silica gel column to obtain KH 28-8. LCMS (ESI, m/z): 638.0 [M+H].
  • Step 8 Dissolve the intermediate KH 28-8 (0.08 g, 0.13 mmol) in acetonitrile (2 mL), add TBD (0.9 M, 0.2 mL) to the reaction system, and stir at room temperature for 2 h. After the reaction is completed, dilute with water (5 mL), adjust the pH to 5-6 with hydrochloric acid (1 M), extract with ethyl acetate (5 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product by Prep-HPLC to obtain KH28.
  • LCMS (ESI, m/z): 624.1 [M+H] + .
  • Example 29 Preparation of (S)-2-(4-(4-((4-cyano-2-methoxybenzyl)oxy)pyrimidin-2-yl)-2,5-difluorobenzyl)-1-((oxetan-2-yl)methyl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxylic acid (KH29) and (S)-2-(4-(2-((4-cyano-2-methoxybenzyl)oxy)pyrimidin-4-yl)-2,5-difluorobenzyl)-1-((oxetan-2-yl)methyl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxylic acid (KH30)
  • Step 4 TBD (0.9 M, 0.72 mL) was added to a solution of KH 29-4 (90 mg, 0.14 mmol) in acetonitrile (1 mL), and the mixture was stirred at room temperature for 1 h. After concentration, the crude product was purified by Prep-HPLC to obtain KH 29. LCMS (ESI, m/z): 614.2 [M+H] + .
  • Step 6 TBD (0.9 M, 0.72 mL) was added to a solution of intermediate KH 30-1 (90 mg, 0.14 mmol) in acetonitrile (1 mL), and the mixture was stirred at room temperature for 1 h. The crude product was concentrated and purified by Prep-HPLC to obtain KH30. LCMS (ESI, m/z): 614.1 [M+H] + .
  • Step 1 Add the raw material KH 31-1 (5 g, 24.5 mmol) and sodium methoxide (1.97 g, 36.4 mmol) in methanol (100 mL) and stir at room temperature for 12 h. After the reaction is complete, add water (50 mL) and ethyl acetate (50 mL x 3) Extraction, washing with saturated brine, drying over anhydrous sodium sulfate, concentration, and purification by silica gel column gave KH 31-2.
  • 1 H NMR 400 MHz, DMSO) ⁇ 8.56 (s, 1H), 7.36 (s, 1H), 3.96 (s, 3H), 3.82 (s, 3H).
  • Step 2 Add lithium borohydride tetrahydrofuran solution (7.5 ml, 14.9 mmol, 2 mol/L) and methanol (0.63 ml, 14.9 mmol) to a solution of the intermediate KH 31-2 (1.5 g, 7.43 mmol) in tetrahydrofuran (15 mL), and stir at 50°C for 1 h.
  • Step 3 Add intermediate KH 31-3 (800 mg, 4.6 mmol) and zinc cyanide (1.076 g, 9.2 mmol) and Pd(dppf)Cl 2 (336 mg, 0.46 mmol) to a DMF (8 mL) solution and stir at 120°C for 3 h. After the reaction is complete, add water (10 mL), extract with ethyl acetate (10 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify with a silica gel column to obtain KH 31-4.
  • LCMS (ESI, m/z): 165.2 [M+H] + .
  • Step 4 To a solution of intermediate KH 31-4 (300 mg, 1.83 mmol) and 2-fluoro-6-bromopyridine (325 mg, 1.83 mmol) in dioxane (6 mL) was added dropwise a solution of potassium tert-butoxide in tetrahydrofuran (2.0 mL, 1 mol/L). Stir at 45°C for 0.5h. After the reaction was completed, water (5mL) was added, extracted with ethyl acetate (5mL x 3), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain crude KH 31-5 which was directly used in the next step.
  • LCMS (ESI, m/z): 319.9 [M+H] + .
  • Step 5 Add potassium carbonate (309 mg, 2.24 mmol) and Pd(dppf)Cl 2 (81.9 mg, 0.112 mmol) to a solution of intermediate Int 31-5 (360 mg, 1.12 mmol) and KH 2-8b (574 mg, 1.12 mmol) in dioxane (6 mL) and water (0.6 ml), and stir at 70°C for 2.5 h. After the reaction is complete, add water (5 mL), extract with ethyl acetate (5 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify on a silica gel column to obtain KH 31-6.
  • LCMS (ESI, m/z): 428.1 [M+H] + .
  • Step 6 Add lithium hydroxide aqueous solution (0.3 mol, 7 ml) to a solution of intermediate KH 31-6 (100 mg, 0.16 mmol) in acetonitrile (2 mL), and stir at 100°C for 5 min. After the reaction is complete, add water (5 mL), add acetic acid aqueous solution dropwise to make the solution acidic, extract with ethyl acetate (5 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain a crude product, which is purified by Prep-HPLC to obtain KH 31.
  • LCMS (ESI, m/z): 601.2[M+H] + .
  • Step 1 Pd(dppf)Cl2 (333 mg, 0.455 mmol) was added to a methanol (100 mL) solution of the raw material KH 32-1 (1 g, 4.55 mmol) and TEA (2.3 g, 22.73 mmol), and stirred at 120 ° C for 12 h under a carbon monoxide pressure atmosphere. After the reaction was completed, 2.5% sodium bicarbonate aqueous solution was added, and ethyl acetate was extracted. The aqueous phase was adjusted to pH 5-6 with 1N hydrochloric acid, and then ethyl acetate was added for extraction.
  • Step 2 Add oxalyl chloride (10.21 g, 80.4 mmol) and catalytic amount of DMF to a solution of intermediate KH 32-2 (800 mg, 4.02 mmol) in dichloromethane (8 mL), and stir at room temperature for 12 h. After the reaction is complete, concentrate to obtain crude KH 32-3, which is directly used for the next step.
  • Step 3 Add intermediate KH 32-3 to a solution of intermediate Int 2c (1.354 g, 4.02 mmol) and TEA (812 mg, 8.04 mmol) in dichloromethane (8 mL) and stir at room temperature for 3 h. After the reaction is complete, add water (10 mL) and dichloromethane (10 mL x 3) to extract, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify on a silica gel column to obtain KH 32-4.
  • LCMS (ESI, m/z): 519.9 [M+H]+.
  • Step 4 Add hydrochloric acid ethyl acetate solution (13 mL, 4 mol/L) to a solution of intermediate KH 32-4 (1.3 g, 2.17 mmol) in ethyl acetate (13 mL) and stir at room temperature for 12 h. After the reaction is complete, filter to obtain KH 32-5.
  • LCMS (ESI, m/z): 399.9 [M+H]+.
  • Step 5 Add DIEA (193.5 mg, 1.5 mmol) to a solution of intermediate KH 32-5 (300 mg, 0.75 mmol) and intermediate Int 3 (163 mg, 0.83 mmol) in dimethyl sulfoxide (3 mL), and stir at 100°C for 12 h. After the reaction is complete, add water (5 mL), extract with ethyl acetate (5 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify on a silica gel column to obtain KH 32-6.
  • Step 6 Add potassium acetate (67 mg, 0.68 mmol) and Pd(dppf)Cl 2 (24.9 mg, 0.034 mmol) to a solution of intermediate KH 32-6 (160 mg, 0.34 mmol) and borane pinacol ester (130 mg, 0.51 mmol) in dioxane (2 mL), and stir at 80°C for 2.5 h. After the reaction is complete, use it directly in the next step.
  • Step 8 Add TBD aqueous solution (0.9 mol, 0.4 ml) to a solution of intermediate KH 32-8 (80 mg, 0.13 mmol) in acetonitrile (2 mL) and stir at room temperature for 1 h. After the reaction is completed, add water (5 mL), add acetic acid aqueous solution to make the solution acidic, extract with ethyl acetate (5 mL ⁇ 3), wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate to obtain a crude product, which is purified by Prep-HPLC to obtain KH32.
  • LCMS (ESI, m/z): 602.0 [M+H]+.
  • Step 1 Add triethylamine (43 g, 424.94 mmol) and Pd(dppf)Cl 2 (3 g, 4.10 mmol) to a methanol (250 mL) solution of the starting material KH33-1 (20 g, 84.38 mmol), and stir the mixture at 120 ° C for 12 h under a carbon monoxide environment.
  • Step 2 Add intermediate Int 2a (8.5 g, 25.22 mmol), 2-chloro-1-methylpyridinium iodide (9.6 g, 37.64 mmol) and DIEA (6.5 g, 50.29 mmol) to a solution of intermediate KH33-2 (5.5 g, 25.46 mmol) in dichloromethane (20 mL), and stir the mixture at room temperature for 12 h.
  • Step 3 Add ethyl acetate (3M, 50 mL) and hydrochloric acid to a solution of intermediate KH33-3 (9 g, 16.75 mmol) in ethyl acetate (50 mL). Stir the mixture at room temperature for 12 h. After the reaction is complete, filter and dry the filter cake to obtain KH33-4 (7 g, yield 96.0%). LCMS (ESI, m/z): 437.1 [M+H] + .
  • Step 4 Add DIEA (14 mL, 77.8 mmol) to a DMSO (70 mL) solution of the intermediate KH33-4 (6.8 g, 15.56 mmol), and stir the reaction at 150 ° C for 2 h. After the reaction is completed, dilute with water (100 mL), extract with ethyl acetate (50 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate. The crude product is purified by silica gel column to obtain KH33-5 (4.7 g, yield 72.7%). LCMS (ESI, m/z): 416.9 [M+H] + .
  • Step 5 Add potassium carbonate (550 mg, 4 mmol) and sodium iodide (60 mg, 0.4 mmol) to a DMF (12 mL) solution of intermediate KH33-5 (800 mg, 2 mmol) and intermediate Int 3 (592 mg, 2.9 mmol), and stir at 50 ° C for 36 h. After the reaction is completed, add water (8 mL) and ethyl acetate (8 mL x 3) for extraction, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify on a silica gel column to obtain KH33-6 (300 mg, yield 32.1%). LCMS (ESI, m/z): 484.8[M+H] + .
  • Step 6 Add potassium acetate (52 mg, 0.534 mmol) and Pd(dppf)Cl 2 (20 mg, 0.027 mmol) to a solution of intermediate KH33-6 (130 mg, 0.267 mmol) and biboronic acid pinacol ester (101 mg, 0.4 mmol) in dioxane (2 mL), and stir at 80° C. for 2.5 h under nitrogen atmosphere to obtain intermediate KH33-7.
  • Step 7 Add water (0.03 mL), intermediate Int 1a (83 mg, 0.267 mmol), potassium carbonate (73.6 mg, 0.543 mmol) and Pd(dppf)Cl 2 (19 mg, 0.0267 mmol) to the reaction solution of intermediate KH33-7, and stir at 90°C for 2.5 h under nitrogen atmosphere. After the reaction is completed, concentrate and purify on a silica gel column to obtain KH33-8 (70 mg, yield 38.7%). LCMS (ESI, m/z): 632.9 [M+H] + .
  • Step 8 Add TBD aqueous solution (0.9M, 0.3mL) to a solution of intermediate KH33-8 (70mg, 0.15mmol) in acetonitrile (2mL) and stir at room temperature for 2h. After the reaction is completed, add water (5mL), add acetic acid aqueous solution dropwise to make the solution acidic, extract with ethyl acetate (5mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product by preparative high performance liquid chromatography to obtain KH33.
  • LCMS (ESI, m/z): 658.8 [M+H] + .
  • Step 1 To a solution of the starting material KH34-1 (136 mg, 0.78 mmol) in 1,4-dioxane (2 mL), add the intermediate KH31-3 (200 mg, 0.78 mmol) and a tetrahydrofuran solution of potassium tert-butoxide (1.5 mL, 1.0 M), and stir the mixture at 45°C for 30 min. After the reaction is complete, concentrate, and purify the crude product by silica gel column to obtain KH34-2 (160 mg, yield 59.0%). LCMS (ESI, m/z): 348.8 [M+H] + .
  • Step 2 Add KH34-2, water (0.02 mL), potassium carbonate (73 mg, 0.543 mmol) and Pd(dppf)Cl 2 (20 mg, 0.03 mmol) to the reaction solution of intermediate KH33-7, and stir at 90°C for 2.5 h under nitrogen atmosphere. After the reaction is completed, add water (5 mL), extract with ethyl acetate (5 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify with silica gel column to obtain KH34-3 (110 mg, yield 61.1%). LCMS (ESI, m/z): 672.9 [M+H] + .
  • Step 3 Add TBD aqueous solution (0.9M, 0.4mL) to a solution of intermediate KH34-3 (100mg, 0.15mmol) in acetonitrile (2mL) and stir at room temperature for 2h. After the reaction is completed, add water (5mL), add acetic acid aqueous solution dropwise to make the solution acidic, extract with ethyl acetate (5mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and obtain a crude product which is purified by preparative high performance liquid chromatography to obtain KH34.
  • LCMS (ESI, m/z): 658.8[M+H] + .
  • Step 1 Add t-BuOK (185 mg, 1.65 mmol) and raw material KH35-1 (230 mg, 1.10 mmol) to a solution of intermediate Int 1e2 (215 mg, 1.32 mmol) in dioxane (10 mL) and react at room temperature for 2 h. After the reaction is complete, add saturated ammonium chloride (10 mL) to quench the reaction, add ethyl acetate (10 mL x 3) to extract the organic phase, wash with saturated brine (20 mL), dry with anhydrous sodium sulfate, and concentrate under vacuum to obtain a crude product. The crude product is purified by normal silica gel to obtain KH35-2 (180 mg, yield 48.8%).
  • Step 2 Dissolve the intermediate KH35-2 (63 mg, 0.19 mmol) in 1,4-dioxane (5 mL) and water (0.5 mL), add the intermediate KH33-7 (120 mg, 0.19 mmol), potassium carbonate (80 mg, 0.57 mmol) and Pd(dppf)Cl 2 (15 mg, 0.02 mmol) to the reaction system, heat to 90 °C under nitrogen atmosphere, and react for 3 h. After the reaction is completed, add water (10 mL) to quench the reaction, add ethyl acetate (10 mL x 3) to extract the organic phase, wash with saturated brine (10 mL), dry with anhydrous sodium sulfate, and concentrate under vacuum to obtain a crude product.
  • Step 3 Add TBD (0.9M, 1mL) to a solution of intermediate KH35-3 (80mg, 0.12mmol) in acetonitrile (5mL) and stir at room temperature for 1h. After the reaction is complete, add saturated brine (10mL) to quench the reaction, add ethyl acetate (10mL x 3) to extract the organic phase, wash with saturated brine (10mL), dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain a crude product. The crude product is purified by reverse C18 to obtain KH35. LCMS (ESI, m/z): 649.0 [M+H] + .
  • Step 1 Add intermediate KH29-3b (106 mg, 0.33 mmol), potassium carbonate (80 mg, 0.57 mmol) and Pd(dppf)Cl 2 (32 mg, 0.04 mmol) to a solution of intermediate KH33-7 (150 mg, 0.28 mmol) in 1,4-dioxane (4 mL) and water (0.4 mL). Stir the mixture at 60°C for 2 h. After the reaction is complete, concentrate the mixture and purify the crude product by silica gel column to obtain KH36-2 (124 mg, yield 68.5%). LCMS (ESI, m/z): 646.1 [M+H] + .
  • Step 2 TBD (0.9M, 0.6mL) was added to a solution of intermediate KH36-2 (124mg, 0.18mmol) in acetonitrile (2mL), and the mixture was stirred at room temperature for 2h. After the reaction was completed, it was diluted with water (5mL), acetic acid (1M) was used to adjust the pH to 5-6, and ethyl acetate (3mL x 3) was used for extraction, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by preparative high performance liquid chromatography to obtain KH36.
  • LCMS ESI, m/z
  • Step 1 Under a nitrogen atmosphere at -78°C, n-butyl lithium (1.7 mL, 2.72 mmol) was added dropwise to a solution of raw material KH37-1 (700 mg, 3.02 mmol) in ether (10 mL), stirred at -78°C for 10 min, and then DMF (0.47 mL, 6.03 mmol) was added dropwise to the reaction solution, and the reaction solution was stirred at -78°C for 20 min under a nitrogen atmosphere.
  • n-butyl lithium 1.7 mL, 2.72 mmol
  • Step 2 Add NaBH 4 (208.86 mg, 5.52 mmol) to a solution of intermediate KH37-2 (500 mg, 2.76 mmol) in ethanol (10 mL) and stir at room temperature for 1.5 h. After the reaction is complete, add water (30 mL), extract with ethyl acetate (30 mL x 3), wash with saturated brine (30 mL), dry with anhydrous sodium sulfate, and concentrate. Obtain a yellow oily crude product KH37-3 (400 mg, 79.12%), which is used directly in the next step.
  • LCMS (ESI, m/z): 184.0 [M+H] + .
  • Step 3 Add compound 2-bromo-6-fluoropyridine (349.42 mg, 1.99 mmol) and cesium carbonate (905.67 mg, 2.78 mmol) to a solution of intermediate KH37-3 (400 mg, 2.18 mmol) in acetonitrile (10 mL), and stir at 95 ° C for 2 h. After the reaction is completed, add water (30 mL), extract with ethyl acetate (30 mL x 3), wash with saturated brine (30 mL), dry with anhydrous sodium sulfate, and concentrate. Purify by silica gel column to obtain KH37-4 (260 mg, yield 38.61%).
  • Step 4 Add intermediate KH37-4 (69.44 mg, 0.205 mmol), K 3 PO 4 (130.39 mg, 0.614 mmol), Pd(dppf)Cl 2 (13.22 mg, 0.0204 mmol) and H 2 O (1 mL) to the reaction solution of intermediate KH33-7, and stir the reaction mixture at 80°C for 2 hours under N 2 atmosphere. After the reaction is completed, add water (15 mL) to dilute, extract with ethyl acetate (10 mL x 3), wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, filter and concentrate. The crude product is purified by silica gel column to obtain KH37-5 (80 mg, yield 58.79%). LCMS (ESI, m/z): 665.1 [M+H] +.
  • Step 5 Add TBD (2 mL, 0.9 M) to a solution of the intermediate KH37-5 (80 mg, 0.120 mmol) in acetonitrile (2 mL), and stir the mixture at room temperature for 30 minutes. After the reaction is completed, adjust the pH of the reaction solution to 5-6 with citric acid, add water (15 mL) to dilute, extract with ethyl acetate (10 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate. The crude product is purified by preparative HPLC to obtain KH37.
  • Step 2 Add the compound 2-bromo-6-fluoropyridine (529.41 mg, 3.01 mmol) to a solution of the intermediate KH38-2 (450 mg, 3.01 mmol) in dioxane (6 mL), and add a solution of potassium tert-butoxide (405.07 mg, 3.61 mmol) in tetrahydrofuran (2 mL) to the reaction solution at room temperature, and stir at 40°C for 1 h. After the reaction is completed, add 1M potassium carbonate aqueous solution (30 mL), extract with ethyl acetate (20 mL x 3), wash with saturated brine (30 mL), dry with anhydrous sodium sulfate, and concentrate.
  • Step 3 Add intermediate KH38-3 (188.28 mg, 0.616 mmol), potassium phosphate (196.19 mg, 0.924 mmol), Pd(dppf)Cl 2 (19.89 mg, 0.0308 mmol) and water (1 mL) to the reaction solution of intermediate KH33-7, and stir at 80°C for 1 h under nitrogen atmosphere. After the reaction is completed, add water (20 mL), extract with ethyl acetate (10 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate. The crude product is purified by silica gel plate to obtain KH38-4 (170 mg, yield 87.45%). LCMS (ESI, m/z): 631.1 [M+H] + .
  • Step 4 Add TBD (3 mL, 0.9 M) to a solution of the intermediate KH38-4 (170 mg, 0.269 mmol) in acetonitrile (3 mL), and stir the mixture at room temperature for 3 hours. LCMS detected the formation of a small amount of product and a large amount of raw materials remaining. TBD (17 mL, 0.9 M) was added and stirred at room temperature overnight. After the reaction was completed, the pH of the reaction solution was adjusted to 5-6 with citric acid, water (10 mL) was added, and ethyl acetate (8 mL x 3) was extracted, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated.
  • Step 1 Add iodomethane (614 mg, 4.32 mmol) and cesium carbonate (2.3 g, 7.05 mmol) to a DMF (10 mL) solution of the raw material KH39-1 (900 mg, 3.61 mmol) and stir at room temperature for 1 h. After the reaction is complete, add water (30 mL) to dilute, extract with ethyl acetate (20 mL x 3), wash with saturated brine (15 mL x 3), dry with anhydrous sodium sulfate, filter, concentrate, and the crude product is purified by normal phase column to obtain KH39-2 (750 mg, yield 78.9%).
  • LCMS (ESI, m/z): No. [M+H] + .
  • Step 2 Add LiOH (340 mg, 14.19 mmol) to a solution of intermediate KH39-2 (750 mg, 2.85 mmol) in methanol (5 mL) and water (1 mL), and stir at room temperature for 4 h. After the reaction is complete, add water (5 mL) to dilute, adjust the pH to 3-4 with dilute hydrochloric acid (1.0 M), extract with ethyl acetate (10 mL x 3), wash with saturated brine (10 mL), dry with anhydrous sodium sulfate, filter, and concentrate to obtain crude KH39-3 (700 mg, yield 99%).
  • LCMS (ESI, m/z): 247.0 [MH] - .
  • Step 3 Add TEA (570 mg, 5.63 mmol) and Pd(dppf)Cl 2 (204 mg, 0.27 mmol) to a solution of the intermediate KH39-3 (700 mg, 2.82 mmol) in methanol (10 mL), and stir overnight at 120°C under a carbon monoxide environment.
  • Step 4 Add intermediate Int 2a (840 mg, 2.49 mmol), 2-chloro-1-methylpyridinium iodide (701 mg, 2.74 mmol) and triethylamine (478 mg, 4.72 mmol) to a DCM (7 mL) solution of intermediate KH39-4 (569 mg, 2.49 mmol), and stir at room temperature for 3 h. After the reaction is complete, concentrate, and the crude product is purified by silica gel column to obtain KH39-5 (500 mg, yield 36.7%). LCMS (ESI, m/z): 612.0 [M+64] + .
  • Step 5 Add ethyl acetate (4M, 8 mL) and hydrochloric acid to a solution of intermediate KH39-5 (500 mg, 0.91 mmol) in ethyl acetate (8 mL), and stir at room temperature overnight. After the reaction is complete, concentrate to obtain the crude product KH39-6 (400 mg, yield 98.0%).
  • LCMS (ESI, m/z): 448.8 [M+H] + .
  • Step 6 Add DIEA (306 mg, 2.36 mmol) to a DMSO (15 mL) solution of the intermediate KH39-6 (350 mg, 0.78 mmol), and stir at 150°C for 2 h. After the reaction is complete, add water (30 mL) to dilute, extract with ethyl acetate (25 mL x 3), wash with saturated brine (30 mL), dry with anhydrous sodium sulfate, filter, concentrate, and the crude product is purified by normal phase column to obtain KH39-7 (90 mg, yield 27.0%).
  • LCMS (ESI, m/z): 426.8 [M+H] + .
  • Step 7 Add intermediate Int 3 (34 mg, 0.17 mmol) and potassium carbonate (39 mg, 0.28 mmol) to a DMF (1.5 mL) solution of intermediate KH39-7 (60 mg, 0.14 mmol), and stir at 50 ° C overnight. After the reaction is completed, add water (30 mL) to dilute, extract with ethyl acetate (20 mL x 3), wash with saturated brine (30 mL), dry with anhydrous sodium sulfate, filter, concentrate, and the crude product is purified by normal phase column to obtain KH39-8 (30 mg, yield 43.1%).
  • LCMS (ESI, m/z): 499.1 [M+H] + .
  • Step 8 Add pinacol borate (23 mg, 0.09 mmol), potassium acetate (12 mg, 0.12 mmol) and Pd(dppf)Cl 2 (5 mg, 0.006 mmol) to a solution of intermediate KH39-8 (30 mg, 0.06 mmol) in 1,4-dioxane (2 mL), and stir at 80°C for 3 h under nitrogen. After the reaction is complete, the system is directly used in the next step (32.8 mg, 100%). LCMS (ESI, m/z): 463.1 [M-82+H] + .
  • Step 9 Add intermediate Int 1a (19 mg, 0.06 mmol), potassium carbonate (17 mg, 0.12 mmol) and Pd(dppf)Cl 2 (5 mg, 0.006 mmol) to a solution of KH39-9 (32.8 mg, 0.06 mmol) in 1,4-dioxane (2 mL) and water (0.2 mL), and stir the mixture at 90°C overnight under nitrogen. After the reaction was completed, the mixture was concentrated and the crude product was purified by normal phase column to obtain KH39-10 (20 mg, 51.7%). LCMS (ESI, m/z): 644.9 [M+H] + .
  • Step 10 TBD (0.9M, 0.5mL) was added to a solution of intermediate KH39-10 (20mg, 0.03mmol) in acetonitrile (1mL), and the mixture was stirred at room temperature for 1.5h.
  • the reaction system was diluted with water (5mL), the pH was adjusted to 3-4 with acetic acid (1M), extracted with ethyl acetate (5mL x 3), washed with saturated brine (10mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by preparative high performance liquid chromatography to obtain KH39.
  • Example 39 A similar method to Example 39 was used, with intermediate KH39-9 and Int 1f-1 as starting materials to obtain compound KH40.
  • LCMS (ESI, m/z): 640.1 [M+H] + .
  • Step 1 Slowly add NaBH 4 (148 mg, 3.91 mmol) to a solution of KH41-1 (500 mg, 2.60 mmol) in tetrahydrofuran (5 mL) and methanol (0.3 mL), and stir the reaction mixture at room temperature for 2 hours. After the reaction is complete, add water to dilute, extract with dichloromethane (20 mL x 3), wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, and concentrate. The crude product is purified by silica gel column to obtain KH41-2 (240 mg, yield 47.50%). LCMS (ESI, m/z): 194.0 [M+H] + .
  • Step 2 TEA (0.52 mL, 3.71 mmol) and MsCl (0.28 mL, 3.71 mmol) were added to a solution of intermediate KH41-2 (240 mg, 1.24 mmol) in dichloromethane (5 mL) in turn, and the reaction solution was stirred at room temperature overnight.
  • TEA 0.52 mL, 3.71 mmol
  • MsCl 0.28 mL, 3.71 mmol
  • Step 3 To a solution of (S)-2-(4-(6-hydroxypyridin-2-yl)-2,5-difluorobenzyl)-1-((oxetan-2-yl)methyl)-3-oxo-2,3-dihydro-1H-indazole-6-carboxylic acid methyl ester (30 mg, 0.059 mmol) in N,N-dimethylformamide (1 mL), intermediate KH41-3 (31.9 mg, 0.117 mmol) and Ag 2 CO 3 (64.69 mg, 0.234 mmol) were added, and the reaction mixture was stirred at 50°C overnight.
  • Step 4 Add TBD (1 mL, 0.9 M) to a solution of the intermediate KH41-4 (12 mg, 0.017 mmol) in acetonitrile (2 mL), and stir the reaction mixture at room temperature overnight. After the reaction is completed, adjust the pH of the reaction solution to 3-4 with citric acid, add water (10 mL), extract with ethyl acetate (10 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate. The crude product is purified by preparative high performance liquid chromatography to obtain KH41. LCMS (ESI, m/z): 673.0 [MH] - .
  • Example 39 A method similar to the ninth and tenth steps of Example 39 was used, and the intermediates KH39-9 and KH38-3 were used as starting materials to obtain the target molecule KH42.
  • LCMS (ESI, m/z): 629.1[M+H] + .
  • the target molecule KH43 was obtained by using a method similar to the ninth and tenth steps of Example 39, with intermediates KH39-9 and KH37-4 as starting materials.
  • Step 1 Slowly add NaBH 4 (168.7 mg, 4.46 mmol) to a solution of the raw material KH44-1 (1 g, 5.86 mmol) in tetrahydrofuran (20 mL) and react at room temperature for 2 h. After the reaction is complete, add saturated ammonium chloride (30 mL) to quench the reaction, add ethyl acetate (30 mL x 3) to extract the organic phase, wash with saturated brine (50 mL), dry with anhydrous sodium sulfate, and concentrate under vacuum to obtain KH44-2 (1.03 g). LCMS (ESI, m/z): 155.2 [M+H-18] + .
  • Step 2 Add t-BuOK (356 mg, 3.17 mmol) to a tetrahydrofuran (10 mL) solution of the intermediate KH44-2 (500 mg, 2.90 mmol), stir at room temperature for half an hour, add compound 4-bromo-2-(methylsulfonyl)pyrimidine (626 mg, 2.64 mmol) and react at 0°C for 2 hours. After the reaction is complete, add saturated ammonium chloride (30 mL) to quench the reaction, add ethyl acetate (20 mL x 3) to extract the organic phase, wash with saturated brine (40 mL), and add anhydrous sulfuric acid.
  • Step 3 KH44-3 (93 mg, 0.28 mmol) was dissolved in 1,4-dioxane (5 mL) and water (0.5 mL), and the intermediate KH2-8b (150 mg, 0.28 mmol), potassium carbonate (78 mg, 0.56 mmol) and Pd(dppf)Cl 2 (22 mg, 0.03 mmol) were added to the reaction system, and the temperature was raised to 90°C under a nitrogen atmosphere for 3 h.
  • Step 4 Add TBD (0.9M, 1mL) to a solution of intermediate KH44-4 (110mg, 0.17mmol) in acetonitrile (5mL), and stir at room temperature for 1h. After the reaction is complete, add saturated brine (10mL) to quench the reaction, add ethyl acetate (10mL x 3) to extract the organic phase, wash with saturated brine (10mL), dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain a crude product, which is purified by reverse C18 to obtain KH44.
  • LCMS (ESI, m/z): 622.9 [M+H] +.
  • Step 1 Add n-butyl lithium (2.56 mL, 4.09 mmol) to a solution of raw material KH45-1 (1 g, 4.31 mmol) in ether (15 mL) at -78 °C and stir at -78 °C for 10 min. Ethyl formate (638.56 mg, 8.62 mmol) was added to the reaction system and stirred at room temperature for 2 h. The reaction was quenched with ammonium chloride solution (30 mL), extracted with ethyl acetate (30 mL x 3), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to obtain KH45-2 (700 mg, crude product). LCMS (ESI, m/z): 181.9 [M+H] + .
  • Step 2 Sodium borohydride (418 mg, 11.04 mmol) was added to an ethanol (10 mL) solution of the intermediate KH45-2 (1 g, 5.52 mmol), and the mixture was stirred at room temperature for 2 h. After the reaction was completed, ammonium chloride (20 mL) was added to quench, extracted with ethyl acetate (30 mL x 3), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the crude product KH45-3 (0.8 g, yield 79.1%).
  • LCMS (ESI, m/z): 183.9 [M+H] + .
  • Step 3 Add triethylamine (164 mg, 1.62 mmol) and methanesulfonyl chloride (149 mg, 1.30 mmol) to a solution of KH45-3 (200 mg, 1.08 mmol) in dichloromethane (3 mL), and stir the mixture at room temperature for 2 h. After the reaction is complete, concentrate to obtain the crude product KH45-4 (250 mg, yield 88.6%).
  • Step 4 Dissolve the intermediate KH2-8b (164.48 mg, 0.32 mmol) in 1,4-dioxane (1 mL) and water (1 mL), and add 6-chloro-2-hydroxypyridine (41.6 mg, 0.32 mmol), K 2 CO 3 (122.88 mg, 0.96 mmol) and Pd(dppf)Cl 2 (21.77 mg, 0.03 mmol) to the reaction system in sequence, and stir the reaction at 90°C for 3 h. After the reaction is completed, concentrate, and purify the crude product by silica gel column to obtain KH45-5 (100 mg, yield 64.97%). LCMS (ESI, m/z): 482.0 [M+H] + .
  • Step 5 Dissolve the intermediate KH45-5 (92 mg, 0.19 mmol) in DMF (3 mL), add the intermediate KH45-4 (100 mg, 0.38 mmol) and silver carbonate (211 mg, 0.76 mmol) to the reaction system, catalyze with sodium iodide, and stir at 50 ° C for 12 h. After the reaction is completed, dilute with water (10 mL), extract with ethyl acetate (5 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and purify the crude product with a silica gel column to obtain KH45-6 (100 mg, yield 40.6%). LCMS (ESI, m/z): 646.9 [M+H] + .
  • Step 6 To a solution of intermediate KH45-6 (100 mg, 0.15 mmol) in acetonitrile (2 mL), TBD (0.9 M, 0.5 mL) was added, and the mixture was stirred at room temperature for 2 h. After concentration, the crude product was purified by preparative HPLC to obtain KH45. LCMS (ESI, m/z): 633.1 [M+H] + .
  • Step 1 Add TEA (8.23 mL, 59.18 mmol) and Pd(dppf)Cl 2 (1.26 g, 1.97 mmol) to a solution of KH46-1 (5 g, 19.73 mmol) in methanol (70 mL). Stir the reaction mixture at 80°C overnight under a carbon monoxide (balloon pressure) atmosphere. After the reaction is complete, concentrate the reaction mixture and dilute it with water (80 mL).
  • Step 2 Add oxalyl chloride (4.11 mL, 49.18 mmol) and DMF (0.18 mL, 2.41 mmol) to a solution of intermediate KH46-2 (5.6 g, 24.08 mmol) in dichloromethane (50 mL) at 0°C, and stir the reaction mixture at room temperature for 30 min. After the reaction is complete, concentrate the reaction solution to obtain KH46-3 (6 g, yield 100%).
  • Step 3 Add the dichloromethane solution of intermediate KH46-3 (6 g, 23.90 mmol) to the dichloromethane (50 mL) solution of intermediate Int 2a (7.31 g, 21.67 mmol) and triethylamine (10.04 mL, 72.23 mmol), and stir the reaction mixture at room temperature for 1 h. After the reaction is completed, add water (80 mL) to dilute, extract with dichloromethane (30 mL x 3), wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate to obtain KH46-5 (11 g, yield 82.81%). LCMS (ESI, m/z): 452.9 [M-100+H] + .
  • Step 4 Add hydrochloric acid-ethyl acetate (50 mL, 4 M) solution to a solution of intermediate KH46-4 (11 g, 19.94 mmol) in ethyl acetate (20 mL), and stir the reaction mixture at room temperature for 6 h. After the reaction is complete, concentrate. The crude product is filtered and purified to obtain KH46-5 (4.9 g, yield 54.42%). LCMS (ESI, m/z): 452.9 [M+H] + .
  • Step 5 A solution of intermediate KH46-5 (4.9 g, 10.85 mmol) in NMP (50 mL) was stirred at 150°C for 4.5 h. After the reaction was completed, water (100 mL) was added to dilute the mixture. Solids precipitated and were filtered. The filter cake was dried to obtain KH46-6 (4.4 g, yield 93.96%).
  • LCMS (ESI, m/z): 433.0 [M+H] + .
  • Step 6 Add K 2 CO 3 (326.61 mg, 2.36 mmol) to a DMF (6 mL) solution of intermediate KH46-6 (340 mg, 0.788 mmol) and Int 3 (311.95 mg, 1.58 mmol), and stir the reaction mixture at 50°C overnight. After the reaction is complete, add water (20 mL) to dilute, extract with ethyl acetate (15 mL x 2), wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, filter and concentrate. The crude product is purified by silica gel column to obtain KH46-7 (130 mg, yield 32.89%). LCMS (ESI, m/z): 502.9 [M+H] + .
  • Step 7 Pd(dppf)Cl 2 (16.52 mg, 0.0259 mmol) and KOAc (76.29 mg, 0.777 mmol) were added to a solution of intermediate KH46-7 (130 mg, 0.259 mmol) and bis(pinacol) borate (131.60 mg, 0.518 mmol) in dioxane (5 mL), and the reaction mixture was stirred at 90°C overnight under N 2 atmosphere. LCMS showed that the reaction was not complete.
  • Step 8 To a solution of intermediate KH46-8 and intermediate KH37-4 (60 mg, 0.177 mmol) in THF (4 mL) and H 2 O (1 mL), K 3 PO 4 (92.83 mg, 0.437 mmol) and Pd(dppf)Cl 2 (9.41 mg, 0.0146 mmol) were added, and the reaction mixture was stirred at 80° C. for 1 hour under N 2 atmosphere. After the reaction was completed, water (15 mL) was added to dilute, and ethyl acetate (10 mL x 3) was used for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The crude product was concentrated. The crude product was purified by silica gel plate to obtain KH46-9 (50 mg, purity 34%, yield 17.12%). LCMS (ESI, m/z): 681.0 [M+H] + .
  • Step 9 Add TBD (1.5 mL, 0.9 M) to a solution of intermediate KH46-9 (50 mg, 0.0734 mmol) in acetonitrile (2 mL), and stir the mixture at room temperature for 2 hours. After the reaction is completed, adjust the pH of the reaction solution to 5-6 with citric acid, add water (10 mL) to dilute, extract with ethyl acetate (8 mL x 3), wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate. The crude product is purified by preparative high performance liquid chromatography to obtain KH46. LCMS (ESI, m/z): 667.0 [M+H] + .
  • Complete culture medium DMEM + 10% fetal bovine serum + 1 ⁇ penicillin-streptomycin + 200 ⁇ g/ml hygromycin
  • reaction plate was centrifuged at 200 g for 30 seconds at room temperature, allowed to stand at 25°C for 1 hour, and data were collected using Envision.
  • Compound PF06882961 was prepared by referring to patent CN110325530A (page 87 of the specification, Example 4A-01).
  • PDE10A1 solution Dilute the PDE10A1 recombinant enzyme stock solution to 15 pg/ ⁇ l with PDE assay buffer and add 20 ⁇ l/well to all compound wells and vehicle control wells. Add 20 ⁇ l PDE assay buffer to the blank control well.
  • binding reagent Take 240 ⁇ l of binding reagent and add it to 23760 ⁇ l of binding reagent diluent, mix well, and add 100 ⁇ l/well to all wells.
  • % inhibition rate (FP V - FP S )/(FP V - FP B ) ⁇ 100%
  • FP S sample fluorescence value
  • FP V vehicle control fluorescence value
  • FP B blank control fluorescence value.
  • hERG human ether-à-go-go related gene potassium channel (IKr, rapidly activating delayed rectifier cardiac potassium current) currents stably expressed in vitro in CHO cell lines was evaluated by manual patch clamp.
  • CHO hERG cells were obtained from Sophion Biosciences (Ballerup, Denmark).
  • test article will first be dissolved in an appropriate solvent and then diluted with ECS at a ratio of 0.1% to form different concentrations of the test article working solution: 0.37, 1.1, 3.3, 10 and 30 ⁇ M.
  • Two cells were measured at each concentration.
  • the hERG current was recorded under the whole-cell patch clamp technique at room temperature.
  • the output signal of the patch clamp amplifier was converted to analog and low-pass filtered at 2.9 kHz.
  • the data was recorded using Patchmaster Pro software.
  • the inhibition percentage of each concentration of the test article was calculated from the recorded current response using the following formula (1-test article/tail peak current recorded after positive control perfusion/tail peak current recorded after vehicle control perfusion (starting current)) ⁇ 100%.
  • the inhibition percentage of all cells recorded was averaged, and the IC 50 value was obtained from the concentration-effect curve by the Hill fitting method.
  • the inhibitory IC 50 of compounds KH03-2, KH13 and KH15-2 on hERG were all greater than 30 ⁇ M.
  • Example 51 Glucose tolerance experiment in genetically engineered hGLP1R mice
  • mice Genetically engineered hGLP1R mice were purchased from Shanghai Nsweeping Model, aged 6-8 weeks, male, weighing 25-30 grams. The quarantine period was 3 days, and routine health examinations were completed by veterinarians. Animals with abnormal performance were eliminated before the experiment. Animals that passed the quarantine were marked with individual animal numbers on their tails and were kept in a SPF-level constant temperature and humidity laminar clean room in an animal center certified by AAALAC, and were kept in single cages (each cage had a cage label indicating the number of animals, sex, strain, receiving time, group, and start time of the experiment).
  • the temperature in the breeding room was 22-25°C, the humidity was 40-70%, and the light was alternating between light and dark for 12 hours (cages: made of polycarbonate, soft corn cob high-pressure sterilized clean bedding, changed twice a week.
  • Feed and drinking water clean-grade mouse feed, purchased from Beijing Keao Xieli Feed Co., Ltd. Drinking water was sterilized by high pressure, and food was irradiated with cobalt 60 rays. Animals can freely take sterile food and drinking water).
  • mice were randomly divided into groups according to their body weight one day before administration. After fasting overnight (17:00-09:00), the blank administration group and other administration groups were administered 30 minutes in advance (the blank administration group used normal saline as the solvent; the other administration groups used 5% DMSO: 20% Solutol: 75% normal saline as the solvent, which was configured into a clear solution).
  • Oral administration oral glucose (2g/kg) was administered after measuring the blood glucose baseline value at 0h, and venous blood was collected from the tail tip of the mice at 15, 30, 60, and 120 minutes. The blood glucose readings of the mice were obtained by using an Accu-Chek Guide blood glucose meter, and the blood glucose change curve was drawn to calculate the AUC area.
  • Example 52 Type II diabetes model of hGLP1R transgenic mice induced by high-fat diet and STZ (HFD-STZ) Experiment
  • mice Male hGLP1R transgenic mice (purchased from Shanghai South Model Organisms Technology Co., Ltd.) aged 10-12 weeks were induced with a high-fat diet (Cat. No. D12492, Research Diets) for 7 weeks, and streptozotocin (STZ) was injected intraperitoneally once a week in the 5th and 6th weeks to induce type II diabetes model; 10-12 week old normal male C57BL/6 mice were normal control group (blank group) and given ordinary diet.
  • a high-fat diet Cat. No. D12492, Research Diets
  • STZ streptozotocin
  • mice were divided into 3 groups according to body weight and fasting blood glucose level, and were given solvent (BID, oral gavage), PF06882961 (positive drug, 15 mg/kg, BID, oral gavage) and test substance KH15-2 (15 mg/kg, BID, oral gavage) respectively; the normal control group was given solvent (BID, oral gavage); each group had 8 mice, except for the normal control group which was given ordinary diet, the other groups were given HFD until the end of the experiment.
  • solvent BID, oral gavage
  • each group had 8 mice, except for the normal control group which was given ordinary diet, the other groups were given HFD until the end of the experiment.
  • the body weight and random blood glucose of the animals were monitored, and an oral glucose tolerance test was performed on all animals at the end of the study.
  • KH15-2 has a good therapeutic effect on HFD+HTZ-induced type II diabetes in hGLP1R mice, and can reduce the body weight, random blood glucose level and fasting blood glucose level of model mice, and enhance glucose tolerance.
  • control compound was prepared according to patent CN114761395A (page 61 of the specification, Example 86).

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Abstract

本发明涉及一种如式I所述的吲唑啉酮类GLP1受体激动剂，所述各取代基的定义如本文中所述；含有该化合物或其盐的组合物以及其制备用于激动GLP-1受体的药物中的用途。

Description

吲唑啉酮类化合物及其制备方法和用途 技术领域
本专利申请涉及药物领域，涉及吲唑啉酮类GLP1受体激动剂，其制备方法及其在医药上的应用。
背景技术
GLP1R(Glucagon-like peptide 1 Receptor)被称为胰高血糖素样肽1受体，属于B类GPCR家族。当人体摄入食物后，肠道L细胞分泌肠促胰素GLP1，该激素以葡萄糖依赖性方式激动GLP1R，促进胰腺β细胞分泌胰岛素，调控体内血糖稳态；同时，导致胃肠道蠕动降低、饱腹感增强、食欲降低。
在2型糖尿病和肥胖症领域，前后已有多款肽类GLP1R激动剂上市。在NASH和神经退行性疾病领域，GLP1R激动剂也展现了临床前/临床上的治疗潜力，相关临床探索也正在积极地开展。但是目前上市的肽类药物大部分均需要皮下注射，影响患者依从性。
PDE10A是一种能同时水解环磷酸腺苷和环磷酸鸟苷的双水解酶，其在大脑中高表达，特别是纹状体中棘神经元。基于文献(Expert Rev Neurother,2017(17),6,553-560；Brain 2015,138(pt 10),3003-3015)的报道，在临床上，以Mardepodect为代表的PDE10A选择性抑制剂意外显示出运动障碍的副作用，这一结果与临床上观察到帕金森和亨廷顿病人纹状体中PDE10A表达降低的现象相一致。这提示我们PDE10A的抑制可能伴随运动方面副作用。
公开的GLP-1受体小分子激动剂专利申请包括WO2009111700A2、WO2010114824A1、WO2018109607A1、WO2019239319A1和WO2018056453A1等。因此，需要提供一种能够口服的、药效好且副作用小的GLP1R激动剂。
发明内容
本发明提供一种能够口服的、药效好、副作用小、溶解性好、生物利用率高的GLP1R激动剂。
本专利申请的目的在于提供一种如式I所示的化合物或其药学上可接受的盐：
其中：
R1选自氢原子、卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、氰基、氨基、硝基、羟基、环烷基、杂环基、芳基和杂芳基，其中所述的烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、环烷基、杂环基、芳基和杂芳基任选地被选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、氰基、氨基、硝基、羟基、羟烷基、环烷基、杂环基、芳基和杂芳基中的一个或多个取代基所取代；
R2选自氢原子、烷基、芳基或杂芳基，其中所述的烷基、芳基或杂芳基任选被选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、氰基、氨基、硝基、羟基、羟烷基、环烷基、杂环基、芳基和杂芳基中的一个或多个取代基所取代，所述环烷基和杂环基任选被卤素、烷基、氰基中的一个或多个取代基所取代；
R3在每次出现时相同或不同，且各自独立地选自氢原子、卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、氰基、氨基、硝基、羟基、环烷基、杂环基、芳基和杂芳基；
R5选自环烷基、杂环基、芳基和杂芳基，其中所述的环烷基、杂环基、芳基和杂芳基任选地被选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、氰基、氨基、硝基、羟基、羟烷基、环烷基、杂环基、芳基和杂芳基中的一个或多个取代基所取代；
X选自N、O或S；
L为直链或支链烷基；
Z0、Z1、Z2、Z3、Z4、Z5和Z6可以相同或不相同，各自独立地选CR4或氮原子；
m、n、p为0-4的整数。
在本专利申请的一些实施例中，R1、R3各自独立地选自氢原子、卤素、C1-6烷基、C1-6烷氧基、卤代C1-6烷基、卤代C1-6烷氧基、氰基、氨基、硝基、羟基、羟C1-6烷基、C3-8环烷基和C3-20杂环基。
在本专利申请的一些实施例中，R2优选被芳基或杂芳基取代的C1-6烷基，所述芳基或杂芳基进一步优选为单环或者稠环形式的芳基或杂芳基，任选地被选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、氰基、氨基、硝基、羟基中的一个或多个取代基所取代。
在本专利申请的一些实施例中，R5优选芳基或杂芳基，所述芳基或杂芳基任选地被烷基、烷氧基、卤代烷基、氰基、卤素所取代。
本发明另一方面提供具有如式II所示的结构化合物或其药学上可接受的盐：
Z0选自C或N；
Z2选自CR4或N，R4选自H或CH3；
Z4、Z5或Z6选自C或N；
R2选自杂环基或C1-6烷基，所述杂环基优选为任选取代的C3-8含氧杂环基，所述C1-6烷基优选被CH2OCH3、取代的C1-6烷基；
R3在每次出现时相同或不同，各自独立地选自氢、F或甲基，n选自1或2的整数；
R5选自取代或未取代的芳基或杂芳基，所述芳基或杂芳基任选被卤素、CN、OC1-6烷基、-C(＝O)C1-6烷基、-C(＝O)-N(C1-6烷基)2、C1-6卤代烷基所取代；
L为CH2，p为1-4的整数。
在一些实施例中，所述芳基或杂芳基选自：
其中X0、X1、X2、X3和X4各自独立的选自C、N、O和S；
优选X0选自C或N；
进一步优选X1、X2、X3和X4至少其中一个是N；
上述芳基或杂芳基任选被H、卤素、氰基、氨基、硝基、羟基、C1-6烷基、C1-6烷氧基、C3-8环烷基、卤代C1-6烷基、卤代C1-6烷氧基、羟C1-6烷基所取代。
本发明另一方面提供具有式III所述的结构的化合物或其药学上可接受的盐：
Z0选自C或N；
Z2选自CR4或N，R4选自H或CH3；
Z4、Z5或Z6选自C或N；
R3选自F，n选自1或2的整数；
R5选自如下芳基或杂芳基：
其中X0选自C或N，
所述芳基或杂芳基可以任选被F、Cl、CN、OCH3、CF3取代；
R6选自-CH2OCH3、R
R10选自H、卤素、CN、OCH3、或CH2CF2。。
本发明另一方面提供具有式IV所示的结构的化合物或其药学上可接受的盐：

Z0选自C或N；
Z4、Z5或Z6选自N或C；
R3是F，n＝2；
R5选自如下基团：
其中X0选自C或N，
R7选自F或甲氧基；R8选自Cl或CN；
R9选自F或H；R11选自Cl或CN；
R12选自卤素或CF3；
R6选自
本发明另一方面提供具有式V所述的结构的化合物或其药学上可接受的盐：
R3选自F，n＝2；
Z6选自C或N；
R5选自如下基团：
其中X0选自C或N，
R7选自F或甲氧基；R8选自Cl或CN；
R9选自F或H；R10选自Cl或CN；
R12选自卤素或CF3；
R6选自优选
本发明另一方面提供具有式VI所述的结构的化合物或其药学上可接受的盐：
其中，
R13选自氢、卤素、C1-3烷基、C1-3卤代烷基或C1-3烷氧基；
Z4、Z5或Z6选自N或C；
X0选自C或N；
R7、R8、R14、R15选自氢、卤素、氰基、C1-3烷基、C1-3卤代烷基或C1-3烷氧基。
本发明另一方面提供具有式VII所述的结构的化合物或其药学上可接受的盐：
其中，
R13选自氢、卤素、C1-3烷基、C1-3卤代烷基或C1-3烷氧基；
Z6选自N或C；
R7、R8选自氢、卤素、氰基、C1-3烷基、C1-3卤代烷基或C1-3烷氧基。
在一些优选的实施例中，R7选自卤素或甲氧基；R8选自卤素或CN。
在另一些优选的实施例中，R7选自F或甲氧基；R8选自Cl或CN。
另一方面，本专利申请提供下述具体化合物:


本专利申请的另一方面涉及一种药物组合物，所述药物组合物含有本专利申请中的通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通式(VI)、通式(VII)及具体化合物，或其可药用的盐，以及一种或多种药学上可接受的载体、稀释剂或赋形剂。
本专利申请进一步涉及通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通式(VI)、通式(VII)及具体化合物，或其可药用的盐，或包含其的药物组合物在制备用于激动GLP-1受体的药物中的用途。
本专利申请进一步涉及通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通式(VI)、 通式(VII)及具体化合物，或其可药用的盐，或包含其的药物组合物在制备用于治疗和/或预防I型糖尿病、II型糖尿病、营养不良相关性糖尿病、糖尿病并发症、肥胖症、高血糖症、葡萄糖耐受不良、心血管疾病、高脂血症、脑梗塞、中风、非酒精性脂肪性肝炎(NASH)、帕金森病、痴呆、胰岛素抗性和肝脏胰岛素抗性的药物中的用途；优选在制备用于治疗和/或预防I型糖尿病、II型糖尿病、肥胖症、糖尿病并发症、非酒精性脂肪性肝炎和心血管疾病的药物中的用途。
本专利申请进一步涉及通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通式(VI)、通式(VII)及具体化合物，或其可药用的盐，或其可药用的盐，或包含其的药物组合物在制备用于治疗和/或预防特发性I型糖尿病、成人隐匿性免疫性糖尿病(LADA)、年青的成年发病型糖尿病(MODY)、妊娠糖尿病、非酒精性脂肪性肝病(NAFLD)、动脉粥样硬化、高血压和冠心病的药物中的用途。
本专利申请还涉及一种激动GLP-1受体的方法，其包括给予所需患者治疗有效量的通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通式(VI)、通式(VII)及具体化合物，或其可药用的盐，或包含其的药物组合物。
本专利申请还涉及一种治疗和/或预防I型糖尿病、II型糖尿病、营养不良相关性糖尿病、糖尿病并发症、肥胖症、高血糖症、葡萄糖耐受不良、心血管疾病、高脂血症、脑梗塞、中风、非酒精性脂肪性肝炎(NASH)、帕金森病、痴呆、胰岛素抗性和肝脏胰岛素抗性的方法；优选I型糖尿病、II型糖尿病、肥胖症、糖尿病并发症、非酒精性脂肪性肝炎和心血管疾病的方法，其包括给予所需患者治疗有效量的通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通式(VI)、通式(VII)及具体化合物，或其可药用的盐，或包含其的药物组合物。
本专利申请还涉及一种治疗和/或预防特发性I型糖尿病、成人隐匿性免疫性糖尿病(LADA)、年青的成年发病型糖尿病(MODY)、妊娠糖尿病、非酒精性脂肪性肝病(NAFLD)、动脉粥样硬化、高血压和冠心病的方法，其包括给予所需患者治疗有效量的通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、、通式(VI)、通式(VII)及具体化合物，或其可药用的盐，或包含其的药物组合物。
本专利申请进一步涉及一种通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通式(VI)、通式(VII)及具体化合物或其可药用的盐，或包含其的药物组合物，其用作药物。
本专利申请还涉及一种通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通式(VI)、通式(VII)及具体化合物，或包含其的药物组合物，其用作GLP-1受体激动剂。
本专利申请进一步涉及一种通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通 式(VI)、通式(VII)及具体化合物，或其可药用的盐，或包含其的药物组合物，其用于治疗和/或预防I型糖尿病、II型糖尿病、营养不良相关性糖尿病、糖尿病并发症、肥胖症、高血糖症、葡萄糖耐受不良、心血管疾病、高脂血症、脑梗塞、中风、非酒精性脂肪性肝炎(NASH)、帕金森病、痴呆、胰岛素抗性和肝脏胰岛素抗性；优选用于治疗和/或预防I型糖尿病、II型糖尿病、肥胖症、糖尿病并发症、非酒精性脂肪性肝炎和心血管疾病。
本专利申请进一步涉及一种通式(I)、通式(II)、通式(III)、通式(IV)、通式(V)、通式(VI)、通式(VII)及具体化合物，或其可药用的盐，或包含其的药物组合物，其用于治疗和/或预防特发性I型糖尿病、成人隐匿性免疫性糖尿病(LADA)、年青的成年发病型糖尿病(MODY)、妊娠糖尿病、非酒精性脂肪性肝病(NAFLD)、动脉粥样硬化、高血压和冠心病。
“糖尿病并发症”是糖尿病或高血糖症引起的并发症，并且其可以是急性复合体或者慢性复合体。术语“急性复合体”包括酮酸症和感染性疾病(例如皮肤感染、软组织感染、胆道系统感染、呼吸系统感染、泌尿道感染)，“慢性复合体”包括例如微血管病(例如肾病、视网膜病)、神经病(例如感觉神经障碍、运动神经障碍、自主神经障碍)和坏疽。主要糖尿病复合体包括糖尿病视网膜病变、糖尿病肾病和糖尿病神经病。
“冠心病”包括心肌梗塞和心绞痛。
“痴呆”包括例如阿尔茨海默氏病、(早发性痴呆)EOD、血管性痴呆和糖尿病性痴呆。
可将活性化合物制成适合于通过任何适当途径给药的形式，通过常规方法使用一种或多种药学上可接受的载体来配制本专利申请的组合物。因此，本专利申请的活性化合物可以配制成用于口服给药、注射(例如静脉内、肌肉内或皮下)给药，吸入或吹入给药的各种剂型。
另一方面，当式I化合物中Z2为N时，在一些实施方案中，本发明提供制备式I化合物的方法(方案1)，
其包括以下步骤：
其中，
PG1为烷基或者苄基，例如甲基、乙基、叔丁基等，优选甲基、乙基；
PG2为烷氧羰基、烷酰基、芳酰基等，例如叔丁氧羰基、苄氧羰基、笏甲氧羰基、烯丙氧羰基、三甲基硅乙氧羰基、三氟乙酰基、邻苯二甲酰基等，优选叔丁氧羰基；
LG1为卤素或取代的磺酸酯基，例如F、Cl、Br、I或三氟甲磺酸酯基，优选F、Cl或三氟甲磺酸酯基；
LG2为卤素或取代的磺酸酯基，例如Cl、Br、I或三氟甲磺酸酯基，优选Br、I或三氟甲磺酸酯基；
LG3为卤素或取代的磺酸酯基，例如Cl、Br、I或三氟甲磺酸酯基，优选Br、I；
LG4为卤素，优选Cl和Br；
其他各基团如上述所定义；
各步骤的反应条件如下：
第一步：原料1制备所得的酰卤与原料2发生亲核取代，或者原料1与原料2直 接缩合生成酰胺中间体3。所述原料1制备酰卤反应在合适的有机溶剂中以及合适的卤化试剂中进行，所述合适有机溶剂包括DCM、THF、DMF等，所述合适的卤化试剂包括草酰氯、二氯亚砜等。所述原料1制备所得的酰卤与原料2发生亲核取代在合适的有机溶剂、温度、碱的存在下反应，所述合适有机溶剂包括DCM、THF、二氧六环等，所述合适的温度范围0-25℃，所述合适的碱包括三乙胺、TIPEA、N-甲基吗啉等。所述原料1与原料2缩合生成酰胺中间体3的反应在合适的缩合剂存在下进行，所述合适的缩合剂包括HATU、PyBOP、DCC等。
第二步：中间体3脱保护形成中间体4。根据PG2保护基的不同，选用通用的脱保护方案。比如，当PG2为Boc时，一般通过酸性水解去除Boc基、当PG2为Cbz时，催化氢解去除Cbz基等。
第三步：中间体4发生分子内的芳香亲核取代，得到异吲唑啉酮中间体5。所述反应在合适的有机溶剂中进行，所述有机溶剂包括正丁醇、DMSO等，优选DMSO；所述反应在合适的反应温度下进行，所述反应温度120-170℃，比如120℃、130℃等，优选150℃。
第四步：中间体5与卤代物原料6发生亲核取代得到中间体7。所述反应在合适的有机溶剂中进行，所述有机溶剂包括DMF、乙醇等，优选DMF；所述反应在合适的反应温度下进行，所述反应温度30-70℃，比如30℃、40℃、50℃等，优选50℃。所述反应在合适的碱的存在下反应，所述碱包括Na2CO3、K2CO3、DIPEA等，优选K2CO3；优选所述反应中间体5与中间体6合适的当量比例为1：1-1：1.5。所述中间体5与所述碱的合适当量比为1：1-1：3，优选1：2。
第五步：中间体7与二硼烷频哪醇酯偶联形成硼酸酯化合物8。所述偶联反应优选在金属催化剂和碱的存在下进行。优选地，所述金属催化剂选自钯金属催化剂，优选[1，1′-双(二苯基膦基)二茂铁]二氯化钯。所述碱优选醋酸钾。所述偶联反应优选溶剂1，4-二氧六环。优选地，所述偶联反应在适合的惰性气氛(例如氮气环境)下进行。优选地，所述偶联反应在适合的温度下进行，所述温度70～90℃，优选80℃。优选地，所述偶联反应进行合适的时间，例如1～4小时，例如2小时。
第六步：中间体8与中间体9发生Suzuki偶联反应形成连芳基中间体10。所述偶联反应优选在金属催化剂、有机磷配体和碱的存在下进行。优选地，所述金属催化剂选自钯金属催化剂，例如三(二亚苄基丙酮)二钯、[1，1′-双(二苯基膦基)二茂铁]二氯化钯、三苯基膦钯、醋酸钯等，优选[1，1′-双(二苯基膦基)二茂铁]二氯化钯。所述有机磷化合物选自BINAP、Xantphos、X-Phos和S-Phos等。所述碱是无机碱或有机碱，例如叔丁醇钠、叔丁醇钾、磷酸钾、碳酸钾、碳酸铯、碳酸钠，优选碳酸铯。优选地，所述偶联反应在 适合的有机溶剂与水组成的混合溶剂中进行，所述混合溶剂可选自DMSO和水、DMF和水、1，4-二氧六环与水、甲苯和水、二甲苯和水等，优选的混合溶剂为1，4-二氧六环和水，二者的优选体积比为10：1～5：1。优选地，所述偶联反应在适合的惰性气氛(例如氮气环境)下进行。优选地，所述偶联反应在适合的温度下进行，所述温度可以是例如80～120℃，优选90℃或100℃。优选地，所述偶联反应进行合适的时间，例如8～24小时，例如12小时。
第七步：酯中间体10经过碱性水解生成终产物11。所述水解反应在合适的混合溶剂中进行，所述混合溶剂包含水和另一种有机溶剂，所述有机溶剂包括甲醇、乙醇、THF等，所述混合溶剂中水与有机溶剂的比例为4：1-1：4；所述反应在合适的碱的存在下进行，所述碱包括K2CO3、LiOH、TBD等，所述中间体10与碱的当量比为1：2-1：3；所述反应在合适的反应温度下进行，所述反应温度40-60℃，比如40℃、50℃等，优选50℃。
当式I化合物中Z0为CH，Z1为CR13，Z2和Z3为N，X为O，Lp为CH2，m＝0,n＝2,R2为两个R3均为F且分别处于苯环2，5位，R5为时，得到如下式VI化合物
在一些实施方案中，本发明提供制备如下式VI化合物的方法，其包括以下步骤：
其中，
PG1为烷基或者苄基，例如甲基、乙基、叔丁基等，优选甲基、乙基；
PG2为烷氧羰基、烷酰基、芳酰基等，例如叔丁氧羰基、苄氧羰基、笏甲氧羰基、烯丙氧羰基、三甲基硅乙氧羰基、三氟乙酰基、邻苯二甲酰基等，优选叔丁氧羰基；
LG1为卤素或取代的磺酸酯基，例如F、Cl、Br、I或三氟甲磺酸酯基，优选F、Cl或三氟甲磺酸酯基；
LG2为卤素或取代的磺酸酯基，例如Cl、Br、I或三氟甲磺酸酯基，优选Br、I或三氟甲磺酸酯基；
LG3为卤素或取代的磺酸酯基，例如Cl、Br、I或三氟甲磺酸酯基，优选Br、I；
LG4为卤素，优选Cl和Br；
其他各基团如上述所定义；
各步骤的反应条件如下：
第一步：原料1制备所得的酰卤与原料2发生亲核取代，或者原料1与原料2直接缩合生成酰胺中间体3。所述原料1制备酰卤反应在合适的有机溶剂中以及合适的卤化试剂中进行，所述合适有机溶剂包括DCM、THF、DMF等，所述合适的卤化试剂包括草酰氯、二氯亚砜等。所述原料1制备所得的酰卤与原料2发生亲核取代在合 适的有机溶剂、温度、碱的存在下反应，所述合适有机溶剂包括DCM、THF、二氧六环等，所述合适的温度范围0-25℃，所述合适的碱包括三乙胺、TIPEA、N-甲基吗啉等。所述原料1与原料2缩合生成酰胺中间体3的反应在合适的缩合剂存在下进行，所述合适的缩合剂包括HATU、PyBOP、DCC等。
第二步：中间体3脱保护形成中间体4。根据PG2保护基的不同，选用通用的脱保护方案。比如，当PG2为Boc时，一般通过酸性水解去除Boc基、当PG2为Cbz时，催化氢解去除Cbz基等。
第三步：中间体4发生分子内的芳香亲核取代，得到异吲唑啉酮中间体5。所述反应在合适的有机溶剂中进行，所述有机溶剂包括正丁醇、DMSO等，优选DMSO；所述反应在合适的反应温度下进行，所述反应温度120-170℃，比如120℃、130℃等，优选150℃。
第四步：中间体5与原料6发生亲核取代得到中间体7。所述反应在合适的有机溶剂中进行，所述有机溶剂包括DMF、乙醇等，优选DMF；所述反应在合适的反应温度下进行，所述反应温度30-70℃，比如30℃、40℃、50℃等，优选50℃。所述反应在合适的碱的存在下反应，所述碱包括Na2CO3、K2CO3、DIPEA等，优选K2CO3；优选所述反应中间体5与中间体6合适的当量比例为1：1-1：1.5。所述中间体5与所述碱的合适当量比为1：1-1：3，优选1：2。
第五步：中间体7与二硼烷频哪醇酯偶联形成硼酸酯化合物8。所述偶联反应优选在金属催化剂和碱的存在下进行。优选地，所述金属催化剂选自钯金属催化剂，优选[1，1′-双(二苯基膦基)二茂铁]二氯化钯。所述碱优选醋酸钾。所述偶联反应优选溶剂1，4-二氧六环。优选地，所述偶联反应在适合的惰性气氛(例如氮气环境)下进行。优选地，所述偶联反应在适合的温度下进行，所述温度70～90℃，优选80℃。优选地，所述偶联反应进行合适的时间，例如1～4小时，例如2小时。
第六步：中间体8与中间体9发生Suzuki偶联反应形成连芳基中间体10。所述偶联反应优选在金属催化剂、有机磷配体和碱的存在下进行。优选地，所述金属催化剂选自钯金属催化剂，例如三(二亚苄基丙酮)二钯、[1，1′-双(二苯基膦基)二茂铁]二氯化钯、三苯基膦钯、醋酸钯等，优选[1，1′-双(二苯基膦基)二茂铁]二氯化钯。所述有机磷化合物选自BINAP、Xantphos、X-Phos和S-Phos等。所述碱是无机碱或有机碱，例如叔丁醇钠、叔丁醇钾、磷酸钾、碳酸钾、碳酸铯、碳酸钠，优选碳酸铯。优选地，所述偶联反应在适合的有机溶剂与水组成的混合溶剂中进行，所述混合溶剂可选自DMSO和水、DMF和水、1，4-二氧六环与水、甲苯和水、二甲苯和水等，优选的混合溶剂为1，4-二氧六环和水，二者的优选体积比为10：1～5：1。优选地，所述偶联反应在适合的惰性气氛(例 如氮气环境)下进行。优选地，所述偶联反应在适合的温度下进行，所述温度可以是例如80～120℃，优选90℃或100℃。优选地，所述偶联反应进行合适的时间，例如8～24小时，例如12小时。
第七步：酯中间体10经过碱性水解生成终产物11。所述水解反应在合适的混合溶剂中进行，所述混合溶剂包含水和另一种有机溶剂，所述有机溶剂包括甲醇、乙醇、THF等，所述混合溶剂中水与有机溶剂的比例为4：1-1：4；所述反应在合适的碱的存在下进行，所述碱包括K2CO3、LiOH、TBD等，所述中间体10与碱的当量比为1：2-1：3；所述反应在合适的反应温度下进行，所述反应温度40-60℃，比如40℃、50℃等，优选50℃。
当式I化合物中Z0为CH，Z1为CR13，Z2和Z3为N，Z4和Z5均为CH，X为O，Lp为CH2，m＝0,n＝2,R2为两个R3均为F且分别处于苯环2，5位，R5为时，得到如下式VII化合物
在一些实施方案中，本发明提供制备如下式VII化合物的方法，其包括以下步骤：
其中，
PG1为烷基或者苄基，例如甲基、乙基、叔丁基等，优选甲基、乙基；
PG2为烷氧羰基、烷酰基、芳酰基等，例如叔丁氧羰基、苄氧羰基、笏甲氧羰基、烯丙氧羰基、三甲基硅乙氧羰基、三氟乙酰基、邻苯二甲酰基等，优选叔丁氧羰基；
LG1为卤素或取代的磺酸酯基，例如F、Cl、Br、I或三氟甲磺酸酯基，优选F、Cl或三氟甲磺酸酯基；
LG2为卤素或取代的磺酸酯基，例如Cl、Br、I或三氟甲磺酸酯基，优选Br、I或三氟甲磺酸酯基；
LG3为卤素或取代的磺酸酯基，例如Cl、Br、I或三氟甲磺酸酯基，优选Br、I；
LG4为卤素，优选Cl和Br；
其他各基团如上述所定义；
各步骤的反应条件如下：
第一步：原料1制备所得的酰卤与原料2发生亲核取代，或者原料1与原料2直接缩合生成酰胺中间体3。所述原料1制备酰卤反应在合适的有机溶剂中以及合适的卤化试剂中进行，所述合适有机溶剂包括DCM、THF、DMF等，所述合适的卤化试剂包括草酰氯、二氯亚砜等。所述原料1制备所得的酰卤与原料2发生亲核取代在合 适的有机溶剂、温度、碱的存在下反应，所述合适有机溶剂包括DCM、THF、二氧六环等，所述合适的温度范围0-25℃，所述合适的碱包括三乙胺、TIPEA、N-甲基吗啉等。所述原料1与原料2缩合生成酰胺中间体3的反应在合适的缩合剂存在下进行，所述合适的缩合剂包括HATU、PyBOP、DCC等。
第二步：中间体3脱保护形成中间体4。根据PG2保护基的不同，选用通用的脱保护方案。比如，当PG2为Boc时，一般通过酸性水解去除Boc基、当PG2为Cbz时，催化氢解去除Cbz基等。
第三步：中间体4发生分子内的芳香亲核取代，得到异吲唑啉酮中间体5。所述反应在合适的有机溶剂中进行，所述有机溶剂包括正丁醇、DMSO等，优选DMSO；所述反应在合适的反应温度下进行，所述反应温度120-170℃，比如120℃、130℃等，优选150℃。
第四步：中间体5与原料6发生亲核取代得到中间体7。所述反应在合适的有机溶剂中进行，所述有机溶剂包括DMF、乙醇等，优选DMF；所述反应在合适的反应温度下进行，所述反应温度30-70℃，比如30℃、40℃、50℃等，优选50℃。所述反应在合适的碱的存在下反应，所述碱包括Na2CO3、K2CO3、DIPEA等，优选K2CO3；优选所述反应中间体5与中间体6合适的当量比例为1：1-1：1.5。所述中间体5与所述碱的合适当量比为1：1-1：3，优选1：2。
第五步：中间体7与二硼烷频哪醇酯偶联形成硼酸酯化合物8。所述偶联反应优选在金属催化剂和碱的存在下进行。优选地，所述金属催化剂选自钯金属催化剂，优选[1，1′-双(二苯基膦基)二茂铁]二氯化钯。所述碱优选醋酸钾。所述偶联反应优选溶剂1，4-二氧六环。优选地，所述偶联反应在适合的惰性气氛(例如氮气环境)下进行。优选地，所述偶联反应在适合的温度下进行，所述温度70～90℃，优选80℃。优选地，所述偶联反应进行合适的时间，例如1～4小时，例如2小时。
第六步：中间体8与中间体9发生Suzuki偶联反应形成连芳基中间体10。所述偶联反应优选在金属催化剂、有机磷配体和碱的存在下进行。优选地，所述金属催化剂选自钯金属催化剂，例如三(二亚苄基丙酮)二钯、[1，1′-双(二苯基膦基)二茂铁]二氯化钯、三苯基膦钯、醋酸钯等，优选[1，1′-双(二苯基膦基)二茂铁]二氯化钯。所述有机磷化合物选自BINAP、Xantphos、X-Phos和S-Phos等。所述碱是无机碱或有机碱，例如叔丁醇钠、叔丁醇钾、磷酸钾、碳酸钾、碳酸铯、碳酸钠，优选碳酸铯。优选地，所述偶联反应在适合的有机溶剂与水组成的混合溶剂中进行，所述混合溶剂可选自DMSO和水、DMF和水、1，4-二氧六环与水、甲苯和水、二甲苯和水等，优选的混合溶剂为1，4-二氧六环和水，二者的优选体积比为10：1～5：1。优选地，所述偶联反应在适合的惰性气氛(例 如氮气环境)下进行。优选地，所述偶联反应在适合的温度下进行，所述温度可以是例如80～120℃，优选90℃或100℃。优选地，所述偶联反应进行合适的时间，例如8～24小时，例如12小时。
第七步：酯中间体10经过碱性水解生成终产物11。所述水解反应在合适的混合溶剂中进行，所述混合溶剂包含水和另一种有机溶剂，所述有机溶剂包括甲醇、乙醇、THF等，所述混合溶剂中水与有机溶剂的比例为4：1-1：4；所述反应在合适的碱的存在下进行，所述碱包括K2CO3、LiOH、TBD等，所述中间体10与碱的当量比为1：2-1：3；所述反应在合适的反应温度下进行，所述反应温度40-60℃，比如40℃、50℃等，优选50℃。
本发明的技术效果一方面在于提供一种具有良好的GLP1R细胞激动活性的化合物。优选的，在本发明的实施方案中，所述化合物在细胞激动活性中EC50小于1000的pM。在一些优选的实施方案中，所述化合物在细胞激动活性中EC50小于100的pM。在另一些优选的实施方案中，所述化合物在细胞激动活性中EC50小于80pM。在另一些优选的实施方案中，所述化合物在细胞激动活性中EC50小于50pM。在另一些优选的实施方案中，所述化合物在细胞激动活性中EC50小于20pM。在另一些优选的实施方案中，所述化合物在细胞激动活性中EC50小于10pM。
本发明的技术效果另一方面在于提供一种具有良好的安全性的化合物。一方面，在PDE10A1的抑制实验中，本发明的化合物显示相对于现有技术的化合物(例如PF06882961)具有更弱的抑制作用。另一方面，在hERG抑制测试中，本发明的化合物显示较高的hERG抑制活性，例如在一些实施方案中，本发明的化合物显示对hERG的抑制IC50均大于30μM。这显示本发明的化合物具有更好的安全性。
本发明的技术效果另一方面在于提供一种溶解性好的化合物。令人惊讶的，本发明所述化合物在模拟空腹肠液溶解度实验中显示出远优于现有化合物的溶解度。具体的，在一些实施方案中，本发明所述化合物具有10-1000μM的溶解度。在一些优选地实施方案中，本发明所述化合物具有50-1000μM的溶解度。在另一些的实施方案中，本发明所述化合物具有50-500μM的溶解度。在另一些的实施方案中，本发明所述化合物具有100-500μM的溶解度。
本发明的技术效果另一方面在于提供一种生物利用度好的化合物。在药代动力学实验中，本发明地化合物在口服暴露量(Cmax和AUC)和生物利用度方面显著优于现有的对照化合物。
术语说明
除非有相反陈述，在说明书和权利要求书中使用的术语具有下述含义。
术语“烷基”、“亚烷基”、“烯基”、“炔基”、“烷氧基”、“环烷基”、“螺环烷基”、“稠环烷基”、“桥环烷基”、“稠杂环基”、“芳基”、“杂芳基”均是取代或未取代的，当被取代时，其可以在任何可使用的连接点上被取代。
术语“烷基”指饱和脂肪族烃基团，其具有1至20个碳原子(即C1-20烷基)。所述烷基优选具有1至12个(例如1、2、3、4、5、6、7、8、9、10、11和12个)碳原子的烷基(即C1-12烷基)，更优选具有1至6个碳原子的烷基(即C1-6烷基)。非限制性实例包括甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基、正庚基、2-甲基己基、3-甲基己基、4-甲基己基、5-甲基己基、2,3-二甲基戊基、2,4-二甲基戊基、2,2-二甲基戊基、3,3-二甲基戊基、2-乙基戊基、3-乙基戊基、正辛基、2,3-二甲基己基、2,4-二甲基己基、2,5-二甲基己基、2,2-二甲基己基、3,3-二甲基己基、4,4-二甲基己基、2-乙基己基、3-乙基己基、4-乙基己基、2-甲基-2-乙基戊基、2-甲基-3-乙基戊基、正壬基、2-甲基-2-乙基己基、2-甲基-3-乙基己基、2,2-二乙基戊基、正癸基、3,3-二乙基己基、2,2-二乙基己基，及其各种支链异构体等。最优选具有1至6个碳原子的低级烷基，非限制性实施例包括甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基等。烷基可以是取代的或非取代的，当被取代时，其可以在任何可使用的连接点上被取代，取代基优选选自D原子、卤素、烷氧基、卤代烷基、卤代烷氧基、环烷基氧基、杂环基氧基、羟基、羟烷基、氰基、氨基、硝基、环烷基、杂环基、芳基、杂芳基中的一个或多个。
术语“亚烷基”指二价烷基，其中烷基如上所定义，其具有1至20个(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20个)碳原子(即C1-20亚烷基)。所述亚烷基优选具有1至12个(例如1、2、3、4、5、6、7、8、9、10、11和12个)碳原子(即C1-12亚烷基)，更优选具有1至6个碳原子的亚烷基(即C1-6亚烷基)。亚烷基的非限制性实例包括但不限于亚甲基(-CH2-)、1,1-亚乙基(-CH(CH3)-)、1,2-亚乙基(-CH2CH2)-、1,1-亚丙基(-CH(CH2CH3)-)、1,2-亚丙基(-CH2CH(CH3)-)、1,3-亚丙基(-CH2CH2CH2-)、1,4-亚丁基(-CH2CH2CH2CH2-)等。 亚烷基可以是取代的或非取代的，当被取代时，其可以在任何可使用的连接点上被取代，取代基优选选自烯基、炔基、烷氧基、卤代烷氧基、环烷基氧基、杂环基氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基和氧代基中的一个或多个。
术语“烯基”指分子中含有至少一个碳碳双键的烷基，其中烷基的定义如上所述，其具有2至12个(例如2、3、4、5、6、7、8、9、10、11和12个)碳原子(即C2-12烯基)。所述烯基优选具有2至6个碳原子(即C2-6烯基)。非限制性的实例包括：乙烯基、丙烯基、异丙烯基、丁烯基等。烯基可以是取代的或非取代的，当被取代时，其优选选自烷氧基、卤素、卤代烷基、卤代烷氧基、环烷基氧基、杂环基氧基、羟基、羟烷基、氰基、氨基、硝基、环烷基、杂环基、芳基和杂芳基中的一个或多个。
术语“炔基”指分子中含有至少一个碳碳三键的烷基，其中烷基的定义如上所述，其具有2至12个(例如2、3、4、5、6、7、8、9、10、11和12个)碳原子。所述炔基优选具有2至6个碳原子(即C2-6炔基)。炔基可以是取代的或非取代的，当被取代时，其优选选自烷氧基、卤素、卤代烷基、卤代烷氧基、环烷基氧基、杂环基氧基、羟基、羟烷基、氰基、氨基、硝基、环烷基、杂环基、芳基和杂芳基中的一个或多个。
术语“烷氧基”指-O-(烷基)，其中烷基的定义如上所述。非限制性的实例包括：甲氧基、乙氧基、丙氧基和丁氧基等。烷氧基可以是取代的或非取代的，当被取代时，其可以在任何可使用的连接点被取代，取代基优选选自氘原子、卤素、烷氧基、卤代烷基、卤代烷氧基、环烷基氧基、杂环基氧基、羟基、羟烷基、氰基、氨基、硝基、环烷基、杂环基、芳基和杂芳基中的一个或多个。
术语“环烷基”指饱和或部分不饱和的单环或多环环状烃取代基，环烷基环具有3至20个(例如3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20个)碳原子，优选具有3至12个碳原子(即3至12元环烷基)，更优选具有3至8个(例如3、4、5、6、7和8个)碳原子(即3至8元环烷基)，最优选具有3至6个碳原子(即3至6元环烷基)。单环环烷基的非限制性实例包括环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、环己二烯基、环庚基、环庚三烯基和环辛基等；多环环烷基包括螺环烷基、稠环烷基和桥环烷基。
术语“螺环烷基”指5至20元，单环之间共用一个碳原子(称螺原子)的多环基团，其可以含有一个或多个双键。优选6至14元，更优选为7至10元(例如7、8、9或10元)。根据环与环之间共用螺原子的数目将螺环烷基分为单螺环烷基或多螺环烷基(如双螺环烷基)，优选为单螺环烷基或双螺环烷基。更优选为4元/4元、4元/5元、4元/6元、5元/5元或5元/6元单螺环烷基。
术语“稠环烷基”指5至20元，系统中的每个环与体系中的其他环共享毗邻的一对碳原子的全碳多环基团，其中一个或多个环可以含有一个或多个双键。优选为6至14元，更优选为7至10元(例如7、8、9或10元)。根据组成环的数目可以分为双环、三环、四环等多环稠环烷基，优选为双环或三环稠环烷基，更优选为3元/4元、3元/5元、3元/6元、4元/4元、4元/5元、4元/6元、5元/4元、5元/3元、5元/5元、5元/6元、6元/3元、6元/4元、6元/5元、6元/6元、6元/7元、7元/5元或7元/6元的双环烷基。
术语“桥环烷基”指5至20元，任意两个环共用两个不直接连接的碳原子的全碳多环基团，其可以含有一个或多个双键。优选为6至14元，更优选为7至10元(例如7、8、9或10元)。根据组成环的数目可以分为双环、三环、四环等多环桥环烷基，优选为双环、三环或四环桥环烷基，更优选为双环或三环桥环烷基。
所述环烷基环包括如上所述的环烷基(包括单环、螺环、稠环和桥环)稠合于芳基、杂芳基或杂环烷基环上，其中与母体结构连接在一起的环为环烷基。
环烷基可以是取代的或非取代的，当被取代时，其可以在任何可使用的连接点上被取代，取代基优选选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、环烷基氧基、杂环基氧基、羟基、羟烷基、氰基、氨基、硝基、环烷基、杂环基、芳基和杂芳基中的一个或多个。
术语“杂环基”指饱和或部分不饱和单环或多环环状取代基，其具有3至20个环原子，其中一个或多个环原子为选自氮、氧和硫的杂原子，所述的硫可任选被氧代(即形成亚砜或砜)，其余环原子为碳。优选具有3至12个(例如3、4、5、6、7、8、9、10、11和12个)环原子，其中1～4个(例如1、2、3和4个)是杂原子(即3至12元杂环基)；更优选具有3至8个环原子(例如3、4、5、6、7和8个)，其中1-3个(例如1、2和3个)是杂原子(即3至8元杂环基)；更优选具有3至6个环原子，其中1～3个是杂原子(即3至6元杂环基)；最优选具有5或6个环原子，其中1～3个是杂原子(即5或6元杂环基)。单环杂环基的非限制性实例包括吡咯烷基、四氢吡喃基、1,2,3,6-四氢吡啶基、哌啶基、哌嗪基、吗啉基、硫代吗啉基和高哌嗪基等。多环杂环基包括螺杂环基、稠杂环基和桥杂环基。
术语“螺杂环基”指5至20元，单环之间共用一个原子(称螺原子)的多环杂环基团，其中一个或多个环原子为选自氮、氧和硫的杂原子，所述的硫可任选被氧代(即形成亚砜或砜)，其余环原子为碳。其可以含有一个或多个双键。优选6至14元，更优选7至10元(例如7、8、9或10元)。根据环与环之间共用螺原子的数目将螺杂环基分为单螺杂环基或多螺杂环基(如双螺杂环基)，优选为单螺杂环基或双螺杂环基。最优选 为3元/5元、3元/6元、4元/4元、4元/5元、4元/6元、5元/5元、5元/6元或6元/6元单螺杂环基。
术语“稠杂环基”指5至20元，系统中的每个环与体系中的其他环共享毗邻的一对原子的多环杂环基团，一个或多个环可以含有一个或多个双键，其中一个或多个环原子为选自氮、氧和硫的杂原子，所述的硫可任选被氧代(即形成亚砜或砜)，其余环原子为碳。优选6至14元，更优选7至10元(例如7、8、9或10元)。根据组成环的数目可以分为双环、三环、四环等多环稠杂环基，优选为双环或三环稠杂环基，更优选为3元/4元、3元/5元、3元/6元、4元/4元、4元/5元、4元/6元、5元/3元、5元/4元、5元/5元、5元/6元、6元/3元、6元/4元、6元/5元、6元/6元、6元/7元、7元/5元或7元/6元双环稠杂环基。
术语“桥杂环基”指5至14元，任意两个环共用两个不直接连接的原子的多环杂环基团，其可以含有一个或多个双键，其中一个或多个环原子为选自氮、氧和硫的杂原子，所述的硫可任选被氧代(即形成亚砜或砜)，其余环原子为碳。优选6至14元，更优选7至10元(例如7、8、9或10元)。根据组成环的数目可以分为双环、三环、四环等多环桥杂环基，优选为双环、三环或四环桥杂环基，更优选为双环或三环桥杂环基。
所述杂环基环包括如上所述的杂环基(包括单环、螺杂环、稠杂环和桥杂环)稠合于芳基、杂芳基或环烷基环上，其中与母体结构连接在一起的环为杂环基，杂环基可以是取代的或非取代的，当被取代时，其可以在任何可使用的连接点上被取代，取代基优选选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、环烷基氧基、杂环基氧基、羟基、羟烷基、氰基、氨基、硝基、环烷基、杂环基、芳基、杂芳基中的一个或多个。
术语“芳基”指具有共轭的π电子体系的6至14元全碳单环或稠合多环(稠合多环是共享毗邻碳原子对的环)基团，优选6至10元，例如苯基和萘基。所述芳基环包括如上所述的芳基环稠合于杂芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为芳基环。
芳基可以是取代的或非取代的，当被取代时，其可以在任何可使用的连接点上被取代，取代基优选选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、环烷基氧基、杂环基氧基、羟基、羟烷基、氰基、氨基、硝基、环烷基、杂环基、芳基和杂芳基中的一个或多个。
术语“杂芳基”指包含1至4个(例如1、2、3和4个)杂原子、5至14个环原子的杂芳族体系，其中杂原子选自氧、硫和氮。杂芳基优选5至10元(例如5、6、7、8、9或10元)，更优选为5元或6元(即5或6元杂芳基)，例如呋喃基、噻吩基、吡啶基、 吡咯基、N-烷基吡咯基、嘧啶基、吡嗪基、哒嗪基、咪唑基、吡唑基、三唑基、四唑基等。所述杂芳基环包括如上述的杂芳基稠合于芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为杂芳基环。杂芳基可以是取代的或非取代的，当被取代时，其可以在任何可使用的连接点上被取代，取代基优选选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、环烷基氧基、杂环基氧基、羟基、羟烷基、氰基、氨基、硝基、环烷基、杂环基、芳基、杂芳基中的一个或多个。
上述环烷基、杂环基、芳基和杂芳基包括从母体环原子上除去一个氢原子所衍生的残基，或从母体的相同或两个不同的环原子上除去两个氢原子所衍生的残基，即“亚环烷基”、“亚杂环基”、“亚芳基”、“亚杂芳基”。
术语“氨基保护基”是指为了使分子其它部位进行反应时氨基保持不变，在氨基上引入的易于脱去的基团。非限制性实施例包含(三甲基硅)乙氧基甲基、四氢吡喃基、叔丁氧羰基(Boc)、乙酰基、苄基、苄氧羰基(Cbz)、烯丙基和对甲氧苄基等。这些基团可任选地被选自卤素、烷氧基或硝基中的1-3个取代基所取代。
术语“羟基保护基”是本领域已知的适当的用于羟基保护的基团。非限制性的实例包括：三甲基硅基(TMS)、三乙基硅基(TES)、三异丙基硅基(TIPS)、叔丁基二甲基硅烷基(TBS)、叔丁基二苯基硅基、甲基、叔丁基、烯丙基、苄基、甲氧基甲基(MOM)、乙氧基乙基、2-四氢吡喃基(THP)、甲酰基、乙酰基、苯甲酰基、对硝基苯甲酰基等。
术语“环烷基氧基”指环烷基-O-，其中环烷基如上所定义。
术语“杂环基氧基”指杂环基-O-，其中杂环基如上所定义。
术语“烷硫基”指烷基-S-，其中烷基如上所定义。
术语“卤代烷基”指烷基被一个或多个卤素取代，其中烷基如上所定义。
术语“卤代烷氧基”指烷氧基被一个或多个卤素取代，其中烷氧基如上所定义。
术语“氘代烷基”指烷基被一个或多个氘原子取代，其中烷基如上所定义。
术语“羟烷基”指烷基被一个或多个羟基取代，其中烷基如上所定义。
术语“卤素”指氟、氯、溴或碘。
术语“羟基”指-OH。
术语“巯基”指-SH。
术语“氨基”指-NH2。
术语“氰基”指-CN。
术语“硝基”指-NO2。
术语“氧代基”或“氧代”指“＝O”。
术语“羰基”指C＝O。
术语“羧基”指-C(O)OH。
术语“羧酸酯基”指-C(O)O(烷基)、-C(O)O(环烷基)、(烷基)C(O)O-或(环烷基)C(O)O-，其中烷基和环烷基如上所定义。
本专利申请的化合物包含其同位素衍生物。术语“同位素衍生物”指结构不同仅在于存在一种或多种同位素富集原子的化合物。例如，具有本专利申请的结构，用“氘”或“氚”代替氢，或者用18F-氟标记(18F同位素)代替氟，或者用11C-，13C-，或者14C-富集的碳(11C-，13C-，或者14C-碳标记；11C-，13C-，或者14C-同位素)代替碳原子的化合物处于本专利申请的范围内。这样的化合物可用作例如生物学测定中的分析工具或探针，或者可以用作疾病的体内诊断成像示踪剂，或者作为药效学、药动学或受体研究的示踪剂。本专利申请的各种氘化形式的化合物是指与碳原子连接的各个可用的氢原子可独立地被氘原子替换。本领域技术人员能够参考相关文献合成氘化形式的化合物。在制备氘代形式的化合物时可使用市售的氘代起始物质，或它们可使用常规技术采用氘代试剂合成，氘代试剂包括但不限于氘代硼烷、三氘代硼烷四氢呋喃溶液、氘代氢化锂铝、氘代碘乙烷和氘代碘甲烷等。氘代物通常可以保留与未氘代的化合物相当的活性，并且当氘代在某些特定位点时可以取得更好的代谢稳定性，从而获得某些治疗优势。
本专利申请化合物可以存在特定的立体异构体形式。术语“立体异构体”是指结构相同但原子在空间中的排列不同的异构体。其包括顺式和反式(或Z和E)异构体、(-)-和(+)-异构体、(R)-和(S)-对映异构体、非对映异构体、(D)-和(L)-异构体、互变异构体、阻转异构体、构象异构体及其混合物(如外消旋体、非对映异构体的混合物)。本专利申请化合物中的取代基可以存在另外的不对称原子。所有这些立体异构体以及它们的混合物，均包括在本专利申请的范围内。可以通过手性合成、手性试剂或者其他常规技术制备光学活性的(-)-和(+)-异构体、(R)-和(S)-对映异构体以及(D)-和(L)-异构体。本专利申请某化合物的一种异构体，可以通过不对称合成或者手性助剂来制备，或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，得到纯的异构体。此外，对映异构体和非对映异构体的分离通常是通过色谱法完成。
本专利申请所述化合物的化学结构中，键“/”表示未指定构型，即如果化学结构中存在手性异构体，键“/”可以为或或者同时包含和两种构型。
本专利申请的化合物可以以不同的互变异构体形式存在，并且所有这样的形式包含在本专利申请的范围内。术语“互变异构体”或“互变异构体形式”是指平衡存在并且 容易从一种异构形式转化为另一种异构形式的结构异构体。其包括所有可能的互变异构体，即以单一异构体的形式或以所述互变异构体的任意比例的混合物的形式存在。非限制性的实例包括：酮-烯醇、亚胺-烯胺、内酰胺-内酰亚胺等。所有的互变异构形式在本专利申请的范围内，且化合物的命名不排除任何互变异构体。
“任选”或“任选地”意味着随后所描述的事件或环境可以但不必发生，该说明包括该事件或环境发生或不发生地场合。例如，“任选被烷基取代的杂环基团”意味着烷基可以但不必须存在，该说明包括杂环基团被烷基取代的情形和杂环基团不被烷基取代的情形。
“取代的”指基团中的一个或多个氢原子，优选为1～5个，更优选为1～3个氢原子彼此独立地被相应数目的取代基取代。本领域技术人员能够在不付出过多努力的情况下(通过实验或理论)确定可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。
“药物组合物”表示含有一种或多种本文所述化合物或其生理学上/可药用的盐或前体药物与其他化学组分的混合物，以及其他组分例如生理学/可药用的载体和赋形剂。药物组合物的目的是促进对生物体的给药，利于活性成分的吸收进而发挥生物活性。
“可药用的盐”是指本专利申请化合物的盐，这类盐用于哺乳动物体内时具有安全性和有效性，且具有应有的生物活性。可以在化合物的最终分离和纯化过程中，或通过使合适的基团与合适的碱或酸反应来单独制备盐。通常用于形成药学上可接受的盐的碱包括无机碱以及有机碱。通常用于形成药学上可接受的盐的酸包括无机酸以及有机酸。
针对药物或药理学活性剂而言，术语“治疗有效量”是指足以达到或至少部分达到预期效果的药物或药剂的用量。治疗有效量的确定因人而异，取决于受体的年龄和一般情况，也取决于具体的活性物质，个案中合适的治疗有效量可以由本领域技术人员根据常规试验确定。
本文所用的术语“药学上可接受的”是指这些化合物、材料、组合物和/或剂型，在合理的医学判断范围内，适用于与患者组织接触而没有过度毒性、刺激性、过敏反应或其他问题或并发症，具有合理的获益/风险比，并且对预期的用途是有效。
本文所使用的，单数形式的“一个”、“一种”和“该”包括复数引用，反之亦然，除非上下文另外明确指出。
当将术语“约”应用于诸如pH、浓度、温度等的参数时，表明该参数可以变化±10％，并且有时更优选地在±5％之内。如本领域技术人员将理解的，当参数不是关键的时， 通常仅出于说明目的给出数字，而不是限制。
附图说明
图1实施例52中各测试化合物的血糖-时间变化曲线
图2实施例52中各测试化合物的血糖-时间积分(AUC)对比图
图3实施例53中各测试化合物的随机血糖-时间变化曲线
图4实施例53中各测试化合物的空腹血糖-时间变化曲线
图5实施例53中各测试化合物的血糖-时间变化曲线
实施例
本文中的缩写具有以下含义：
缩写     含义
TLC     薄层色谱法
LC-MS      液相色谱-质谱联用
DMF     N，N-二甲基甲酰胺
DMSO      二甲基亚砜
EA         乙酸乙酯
PE      石油醚
THF     四氢呋喃
LDA    二异丙基氨基锂
EDCI     1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐
HOBT    1-羟基苯并三唑
Pd(dppf)Cl2    [1，1’-双(二苯基膦基)二茂铁]二氯化钯
SFC     超临界流体色谱
ee      对映体过量百分数
PPA        多聚磷酸
DCE     1,2-二氯乙烷
NBS     N-溴代丁二酰亚胺
AIBN    偶氮二异丁腈
TBD     157-三叠氮双环(4.4.0)癸-5-烯
Solutol       2-羟乙基-12-羟基十八酸酯
以下实施例中记载的化合物的结构通过核磁共振氢谱(1H-NMR)或质谱(MS)来确证。
1H-NMR的测定仪器为Bruker 400MHz核磁共振仪，测定溶剂为氘代甲醇(CD3OD)、氘代氯仿(CDCl3)或六氘代二甲基亚砜(DMSO-d6)，内标为四甲基硅烷(TMS)。化学位移(δ)以百万分之一(ppm)为单位给出。
质谱(MS)的测定仪器为Agilent(ESI)质谱仪，型号为Agilent 6120B。
薄层色谱法(TLC)使用Merck产的铝板(20×20cm)进行，薄层制备色谱法采用GF254(0.4～0.5mm)硅胶板进行。
反应的监测采用薄层色谱法(TLC)或液相色谱-质谱联用(LC-MS)，使用的展开剂体系包括二氯甲烷和甲醇体系、正己烷和乙酸乙酯体系以及石油醚和乙酸乙酯体系。根据要分离的化合物的极性不同对展开剂体系进行调节(通过调节溶剂的体积比或者加入三乙胺等进行)。
制备高效液相色谱法所使用的仪器型号：Agilent 1260，色谱柱：Waters XBridge Prep C18OBD(19mm×150mm×5.0μm)；色谱柱温：25℃；流速：20.0mL/min；检测波长：214nm；洗脱梯度：(0min：10％A，90％B；16.0min：90％A，10％B)；流动相A：100％乙腈；流动相B：0.05％碳酸氢铵水溶液。
除非特别指出，反应温度为室温(20℃～30℃)。
实施例中所使用的试剂购自Acros Organics、Aldrich Chemical Company、上海特伯化学科技有限公司等。
中间体：
共用中间体的制备
中间体4-(((6-溴吡啶-2-基)氧基)甲基)-3-氟氰基苯(Int 1a)的制备：
向原料2-氟-6-溴吡啶(10g,57.14mmol)的THF(100mL)溶液中，加入2-氟4-氰基苄醇(8.59g,57.14mmol)和叔丁醇钾(7.68g,68.57mmol)，40℃搅拌1h，监测反应完成后，加入150mL水，乙酸乙酯(100mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥后过滤,滤液浓缩，粗品经硅胶层析柱纯化得Int 1a。LCMS(ESI,m/z):307.0[M+H]+.
中间体2-溴-6-((4-氯-2-氟苯并呋喃-7-基)甲氧基)吡啶(Int 1b)的制备
第一步:向原料Int 1b1(10g,69.93mmol)和溴代乙醛缩二乙醇(16.55g,83.92mmol)的DMF(100mL)溶液中，加入碳酸钾(14.50g,104.89mmol)，于120℃搅拌过夜。TLC监控反应完毕后，加入100mL水，乙酸乙酯(100mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥后过滤,滤液浓缩得到粗品Int 1b2，该中间体无需纯化直接用于下一步反应。
第二步:向中间体Int 1b2(18g,69.56mmol)的DCE(180mL)溶液中，加入PPA30mL，氮气保护下，于80℃搅拌过夜。TLC监测反应完毕后，加入水200mL，乙酸乙酯(200mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥，过滤，浓缩后的滤液经硅胶柱色谱纯化,得到Int 1b3。
第三步:向中间体Int 1b3(7.8g,46.70mmol)的DCE(80mL)溶液中，加入NBS(10.00g,56.04mmol)和AIBN(1.54g,9.34mmol)，升温至72℃搅拌过夜。反应完毕后，加入水100mL，乙酸乙酯(100mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥,过滤，浓缩的滤液经硅胶柱纯化,得到Int 1b4。1H NMR(400MHz,DMSO)δ8.17(d,J＝2.2Hz,1H),7.41(d,J＝8.0Hz,1H),7.29(d,J＝8.0Hz,1H),7.01(d,J＝2.2Hz,1H),4.90(s,2H).
第四步:向中间体Int 1b4(2g,8.13mmol)的DMF(20mL)溶液中，加入乙酸钾(8g,81.30mmol)，室温下搅拌3h。TLC监测反应完毕，加入水20mL，乙酸乙酯(20mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥,过滤，滤液浓缩得到粗品Int 1b5，该产品无需纯化直接用于下一步。
第五步:向中间体Int 1b5(1.8g,8.00mmol)的THF(20mL)溶液中，加入甲醇钠(0.86g,16.00mmol)，室温搅拌1h。反应完毕，加入水100mL，乙酸乙酯(100mL x 3)萃取，饱和食盐水洗涤有机相后，无水硫酸钠干燥,过滤，浓缩的滤液经硅胶柱色 谱纯化,得到Int 1b6。1H NMR(400MHz,DMSO)δ8.11(d,J＝2.2Hz,1H),7.34(q,J＝8.0Hz,2H),7.00(d,J＝2.2Hz,1H),5.40(s,1H),4.79(s,2H).
第六步:向中间体Int 1b6(0.9g,4.91mmol)和2-氟-6-氯吡啶(0.62mg,4.91mmol)的二氧六环和水(Dioxane:H2O＝10:1，10mL)溶液中，加入叔丁醇钾(1.12g,9.84mmol)的THF(3ml)溶液，45℃下搅拌0.5h。反应完毕，加入10mL水，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤有机相后，无水硫酸钠干燥、过滤，浓缩后的滤液经硅胶柱层析纯化,得到Int 1b7。LCMS(ESI,m/z):339.9[M+H]+.1H NMR(400MHz,DMSO)δ8.17(d,J＝2.2Hz,1H),7.73-7.64(m,1H),7.46(d,J＝8.0Hz,1H),7.38(d,J＝8.0Hz,1H),7.27(d,J＝7.5Hz,1H),7.06(d,J＝2.2Hz,1H),6.93(d,J＝8.2Hz,1H),5.59(s,2H).
第七步:向中间体Int 1b7(1.0g,2.95mmol)的超干四氢呋喃(10mL)溶液中，加入LDA(2M,2.2mL,4.40mmol)，-60℃搅拌1h后，再加入N-氟代双苯磺酰胺(9.3g,29.50mmol)，-60℃继续搅拌1h后，升温至室温再搅拌1h。反应完毕，加入水(10mL)，用乙酸乙酯(10mL x 3)萃取，萃取的有机相经饱和食盐水洗涤、无水硫酸钠干燥,过滤，浓缩的滤液经硅胶柱层析纯化,得到Int 1b。LCMS(ESI,m/z):357.8[M+H]+.1H NMR(400MHz,DMSO)δ7.74-7.62(m,1H),7.44(q,J＝8.2Hz,2H),7.27(d,J＝7.5Hz,1H),6.94(d,J＝8.2Hz,1H),6.52(d,J＝6.4Hz,1H),5.53(s,2H).
中间体6-(((6-溴吡啶-2-基)氧基)甲基)-1-(2,2-二氟乙基)-1H-吲唑(Int 1c)的制备
第一步：向原料Int 1c-1(1g,5.68mmol)的DMF(10mL)溶液中，加入1,1-二氟-2-碘代乙烷(1.1g,5.73mmol)和碳酸铯(3.7g,11.35mmol)，室温下搅拌16h，反应完毕，浓缩，粗品经正相色谱纯化(PE:EA＝5:1)得Int 1c-2。LCMS(ESI,m/z):241.0[M+H]+.
第二步：向中间体Int 1c-2(725mg,3.02mmol)的THF(8mL)的溶液中加入甲醇(194mg,6.06mmol)和硼氢化锂(3mL,6.06mmol)，混合物在50℃下搅拌12h。反应完毕，加入饱和氯化铵(10mL)淬灭，乙酸乙酯(15mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩得粗品Int 1c-3。LCMS(ESI,m/z):213.0[M+H]+.
第三步：向中间体Int 1c-3(733mg,3.46mmol)的二氧六环(10mL)溶液中，加入2-氟-6-溴吡啶(608mg,3.45mmol)和叔丁醇钾(484mg,4.32mmol)的四氢呋喃(3mL)溶液，反应在40℃下搅拌30min。反应完毕后，加入饱和氯化铵(20mL)溶液淬灭，加入水(20mL)稀释反应体系，乙酸乙酯(15mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩得粗品Int 1c。LCMS(ESI,m/z):369.9.
中间体4-(((6-溴-3-氟-吡啶-2-基)氧基)甲基)-3-氟氰基苯(Int 1d)的制备
向3-氟-2,6-二溴吡啶(400mg,1.57mmol)和2-氟4-氰基苄醇(236.86mg,1.57mmol)的二氧六环(5mL)溶液中，加入叔丁醇钾(351.68mg,3.14mmol)的四氢呋喃(1mL)溶液，45℃搅拌30min。反应完毕，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化,得到Int 1d。LCMS(ESI,m/z):325.0[M+H]+.1H NMR(400MHz,DMSO)δ7.94(d,J＝10.2Hz,1H),7.78-7.72(m,3H),7.32(dd,J＝8.2,2.7Hz,1H),5.52(s,2H).
中间体4-(((6-溴吡啶-2-基)氧基)甲基)-3-甲氧基苯氰(Int 1e)的制备
第一步：向原料Int 1e1(950mg,5.36mmol)的THF(10mL)溶液中，加入二甲硫醚硼烷(16mL,32.17mmol)，室温下搅拌1h。反应完毕，加入水(10mL)，乙酸乙 酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩得中间体Int 1e2。该粗品直接用于下一步。LCMS(ESI,m/z):146.1[M+H]+.1H NMR(400MHz,DMSO)δ7.55(d,J＝7.7Hz,1H),7.47-7.38(m,2H),5.30(t,J＝5.6Hz,1H),4.53(d,J＝5.8Hz,2H),3.84(s,3H).
第二步：向中间体Int 1e2(510mg,3.13mmol)和2-氟-6-溴吡啶(550.88mg,3.13mmol)的二氧六环(6mL)溶液中，加入t-BuOK(420.51mg,3.75mmol)的THF(2mL)溶液，45℃搅拌0.5h。反应完毕，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到Int 1e。LCMS(ESI,m/z):318.9[M+H]+.
中间体2-乙撑基-3-(叔丁基二甲基硅氧基)碘丙烷(Int 1f)的制备
第一步：在0℃下向原料Int 1f1(5.0g,48.95mmol)的DCM(100mL)溶液中加入咪唑(5g,73.44mmol)，滴加TBSCl(7.35g,48.95mmol)的DCM(40mL)溶液，室温搅拌3小时。反应结束后，降到0℃，加入水(50ml),乙酸乙酯(120mL x 3)萃取,硫酸钠干燥，浓缩，粗产品经硅胶柱纯化后得到Int 1f2。1H NMR(400MHz,CDCl3)δ3.55(s,2H),3.48(d,J＝16.3Hz,2H),0.86-0.83(m,9H),0.47-0.42(m,2H),0.41-0.34(m,2H),0.03--0.02(m,6H).
第二步：向中间体Int 1f2(4.3g,19.81mmol)的DMF(100mL)溶液中加入三苯基膦(6.24g，23.78mmol)，咪唑(1.62g,23.78mmol)和碘(5.54g，21.79mmol)，-20℃搅拌2h，反应结束后加入水(200ml),用乙醚(120mL x 3)萃取,有机相用硫酸钠干燥，浓缩，粗产品经硅胶柱纯化后得到Int 1f。1H NMR(400MHz,CDCl3)δ3.45(s,2H),3.24(s,2H),0.84-0.82(m,9H),0.54-0.49(m,2H),0.02-0.06(m,8H).
中间体Int 1f-1、Int 1f-2、Int 1f-3、Int 1f-4的制备
以市售的苄醇为原料，采用Int 1a类似的合成方法，分别制备得到如下中间体。
表1

中间体2-(((6-溴吡啶-2-基)氧基)甲基)-5-氯-N,N-二甲基苯甲酰胺(Int 1f-5)的制备
第一步：向原料Int 1f-5-1中(850mg,5.04mmol)加入二甲胺的四氢呋喃溶液(8.5mL)，70℃搅拌12h。反应完毕，浓缩。经硅胶柱纯化得到Int 1f-5-2。LCMS(ESI,m/z):214.0[M+H]+.1H NMR(400MHz,DMSO)δ7.55-7.43(m,2H),7.26(d,J＝2.2Hz,1H),5.28(t,J＝5.6Hz,1H),4.38(d,J＝5.6Hz,2H),2.97(s,3H),2.75(s,3H).
第二步：向中间体Int 1f-5-2(600mg,2.80mmol)和2-氟-6-溴吡啶(493.46mg,2.80mmol)的二氧六环(6mL)溶液中，加入t-BuOK(392.52mg,3.50mmol)的THF(2mL)溶液，45℃搅拌0.5h。反应完毕，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到Int 1f-5。LCMS(ESI,m/z):318.9[M+H]+.1H NMR(400MHz,DMSO)δ7.69(t,J＝7.8Hz,1H),7.54-7.59(m,1H),7.48-7.52(m,1H),7.39-7.42(m,1H),7.24-7.28(m,1H),6.89(d,J＝8.2Hz,1H),5.25(s,2H),2.93(s,3H),2.81(s,3H).
中间体2-(4-溴-2,5-二氟苄基)肼-1-羧酸叔丁醇酯(Int 2a)的制备
第一步：向4-溴-2,5-二氟苯甲醛Int 2a-1(5g,22.7mmol)的THF(50mL)溶液中，加入肼基甲酸叔丁酯(3.6g,27.2mmol)和氰基硼氢化钠(2.8g,45.4mmol)，于50℃下搅拌3h，反应完毕，浓缩反应液，粗品Int 2a-2直接用于下一步。
第二步：向中间体Int 2a-2(7.6g,22.7mmol)的甲醇(60mL)溶液中，加入硼氢化钠(1.7g,45.4mmol)，反应在室温下搅拌1h。反应完毕，加入水(100mL)稀释反应体系，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤有机相，无水硫酸钠干燥，过滤，滤液浓缩后经硅胶柱层析纯化得Int 2a。LCMS(ESI,m/z):378.2[M+H+MeCN]+.
中间体2-(4-溴-2-氟苄基)肼-1-羧酸叔丁醇酯(Int 2b)和2-(4-溴-3-氟苄基)肼-1-羧酸叔丁醇酯(Int 2e)的制备
采用Int2a类似的方法，以不同的芳香醛和肼基甲酸叔丁酯为起始物料，得到中间体Int 2b、Int 2e如下：
表2
中间体2-(4-溴-2-氟-5-甲基苄基)肼-1-羧酸叔丁醇酯(Int 2c)的制备
第一步：向原料Int 2c-1(5g,21.46mmol)的THF(50mL)溶液中加入硼烷二甲硫醚(21.4mL,2M)，室温搅拌过夜。反应完后加入HCl(1N,10mL)淬灭反应后，过滤反应液，浓缩滤液除去四氢呋喃，加入水(30mL)，用乙酸乙酯萃取(3×50mL)，有机相干燥、过滤，减压蒸干溶剂得到Int 2c-2粗品。1H NMR(400MHz,CDCl3)δ7.16-7.23(m,2H),4.61(s,2H),2.29(s,3H).
第二步：中间体Int 2c-2(4.3g,19.63mmol)溶于二氯甲烷(100mL)中，在0℃下，加入戴斯马丁氧化剂(16.65g,39.26mmol)，室温搅拌过夜，反应完后用饱和碳酸氢钠溶液(80mL)淬灭反应，二氯甲烷(3×100mL)萃取，用10％硫代硫酸钠溶液洗涤有机相，硫酸钠干燥后过滤，浓缩除去溶剂，粗品经硅胶柱层析色谱纯化得Int2c-3。1H NMR(400MHz,CDCl3)δ10.29(s,1H),7.72(dd,J＝7.3,0.4Hz,1H),7.42(d,J＝9.6Hz,1H),2.42(s,3H).
第三步：中间体Int 2c-3(3.187g,14.68mmol)和叔丁氧羰基肼(2.91g,22.03mmol)溶于甲醇中(50mL)，50℃反应2h，反应完后减压蒸干溶剂，粗品经硅胶柱层析色谱纯化得Int 2c-4。LCMS(ESI，m/z)：275.0[M+H-56]+。1H NMR(400MHz,CDCl3)δ7.99(d,J＝12.6Hz,2H),7.91(d,J＝7.6Hz,1H),7.26-7.28(m,1H),2.35(s,3H),1.54(s,9H).
第四步：中间体Int 2c-4(4.70g,14.20mmol)溶于THF(80mL),氮气保护下于-78℃下滴加DIBAL-H(59.4mL,1.5M)的四氢呋喃溶液，滴加完后逐渐升温到室温反应2h，反应结束后滴加甲醇(10mL)淬灭反应。浓缩反应液，向残留物中加入饱和酒石酸钠钾(300mL)溶液，搅拌至溶液澄清，再加入乙酸乙酯(200mL×3)萃取，有机相合并后用食盐水洗涤，无水硫酸钠干燥有机相，过滤，滤液浓缩后经硅胶柱层析色谱纯化得Int 2c。LCMS(ESI，m/z)：279.0[M+H-56]+。1H NMR(400MHz,CDCl3)δ7.15-7.18(m,1H),7.07-7.12(m,1H),5.96(s,1H),4.15(s,1H),3.90(d,J＝4.8Hz,2H),2.27(s,3H),1.38(s,9H).
中间体2-(4-溴-2,6-二氟苄基)肼-1-羧酸叔丁醇酯(Int2d)的制备
第一步：向原料Int 2d-1(5g，24.63mmol)的四氢呋喃(50mL)溶液中加入肼基甲酸叔丁酯，于50℃下反应2h。反应完全后，待反应液恢复至室温，真空减压浓缩得Int 2d-2。LCMS(ESI，m/z)：278.9[M-55]+。
第二步：氮气保护下，中间体Int 2d-2(9g，26.85mmol)的四氢呋喃(100mL)溶液降温至于-78℃，再加入DIBAL-H(100mL，161.12mmol)，-78℃下反应1h。反应完全后，将反应液倒入水中，加入乙酸乙酯萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥，过滤，真空减压浓缩滤液得粗品，经硅胶柱层析色谱纯化得Int 2d。LCMS(ESI，m/z)：281.0[M-55]+。
中间体(S)-2-碘甲基氧杂环丁烷(Int 3)的制备
第一步:将原料三甲基碘化亚砜(161g,732mmol)和叔丁醇钾(82g,732mmol)溶于叔丁醇(600mL)溶液中，60℃搅拌0.5h后升温到80℃,滴加原料Int 3-1(60g,366mmol)，搅拌2h。反应完毕，加入水(200mL)，乙酸乙酯(200mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥,过滤，滤液浓缩所得粗品经硅胶柱层析色谱纯化得到Int 3-2。1H NMR(400MHz,DMSO)δ7.38-7.28(m,5H),4.88-4.82(m,1H),4.56-4.48(m,3H),4.436-4.384(m,1H),3.62(dd,J＝11.2,5.6Hz,1H),3.53(dd,J＝11.2,3.4Hz,1H),2.64-2.54(m,1H),2.47-2.41(m,1H).
第二步:向中间体Int 3-2(25g,140.4mmol)的甲醇(350mL)溶液中，加入钯碳(10g)，氢气氛围下于50℃搅拌12h。反应完毕,过滤，滤液浓缩，硅胶色谱柱纯化得到Int 3-3。1H NMR(400MHz,DMSO)δ4.81(t,J＝5.6Hz,1H),4.71-4.61(m,1H),4.52-4.42(m,1H),4.40-4.35(m,1H),3.48(t,J＝5.0Hz,2H),2.56-2.49(m,1H), 2.45-2.37(m,1H).
第三步:向50mL二氯甲烷中，加入咪唑(12.85g,189mmol)、三苯基磷(24.65g,94.1mmol)与碘单质(23.9g,94.1mmol)，0℃搅拌0.5h。恢复到室温后，滴加中间体Int 3-3(6.44g,73.1mmol)，室温搅拌12h。反应完毕，加入水(200mL)、乙酸乙酯(200mL x 3)萃取，饱和硫代硫酸钠水与饱和食盐水洗涤有机相，无水硫酸钠干燥有机相后,过滤，浓缩所得粗品经硅胶柱层析色谱纯化得到Int 3。1H NMR(400MHz,DMSO)δ4.74-4.63(m,1H),4.44-4.32(m,2H),3.52-3.41(m,2H),2.75-2.59(m,1H),2.36-2.19(m,1H).比旋度[α]D 20＝-52.859(c＝1.0272,MeOH,ee 100％)
实施例1：2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2-氟苄基)-3-(2-甲氧乙基)-3-甲基-1-氧代异吲哚啉-5-羧酸(KH01)、(R)-2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2-氟苄基)-3-(2-甲氧乙基)-3-甲基-1-氧代异吲哚啉-5-羧酸(KH01-1)和(S)-2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2-氟苄基)-3-(2-甲氧乙基)-3-甲基-1-氧代异吲哚啉-5-羧酸(KH01-2)的合成
第一步：向2-溴甲基-4-溴苯甲酸甲酯KH 1-1(5g,16.23mmol)的DMF(50mL)溶液中分别加入对甲氧基苄胺(2.23g,16.23mmol)和三乙胺(3.28g,32.46mmol),于室温搅拌2h。反应完毕后，加入水100mL，EA萃取(100mL*3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，过滤，浓缩后的滤液经硅胶柱层析色谱分离得KH 1-2。 LCMS(ESI,m/z):332.0[M+H]+.1H NMR(400MHz,CDCl3)δ7.73(t,J＝7.2Hz,1H),7.59(dd,J＝8.2,1.4Hz,1H),7.51(s,1H),7.33(d,J＝8.4Hz,2H),6.83(d,J＝8.6Hz,2H),4.82(d,J＝15.4Hz,1H),4.43(t,J＝14.8Hz,1H),3.79(d,J＝7.4Hz,3H),2.98(s,3H),2.74-2.60(m,2H),2.33-2.20(m,1H),2.18-2.07(m,1H),1.34(s,3H).
第二步：向KH 1-2(1.5g,4.53mmol)的DMF(15mL)溶液中加入氢化钠(362mg,9.06mmol)，混合物于80℃搅拌30min后，冷却至室温，向混合物中加入碘甲烷(772mg,5.44mmol),再室温搅拌1h。反应完毕，加入15mL水，EA萃取(15mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，过滤，浓缩的粗品经硅胶柱层析色谱纯化得KH1-3。LCMS(ESI,m/z):346.0[M+H]+.
第三步：向KH 1-3(680mg,1.97mmol)的DMF(7mL)溶液中加入氢化钠(158mg,3.94mmol)和2-溴乙基甲基醚(544mg,3.94mmoL),混合物于80℃搅拌1h。反应完毕后，加入水(10mL)，EA(10mL*3)萃取，饱和食盐水洗涤有机相，无水硫酸钠干燥，过滤，滤液浓缩，粗品经硅胶柱层析纯化得KH 1-4。1H NMR(400MHz,CDCl3)δ7.73(t,J＝7.2Hz,1H),7.59(dd,J＝8.2,1.4Hz,1H),7.51(s,1H),7.33(d,J＝8.6Hz,2H),6.83(d,J＝8.6Hz,2H),4.82(d,J＝15.4Hz,1H),4.43(t,J＝14.8Hz,1H),3.79(d,J＝7.4Hz,3H),2.98(s,3H),2.74-2.60(m,2H),2.33-2.20(m,1H),2.18-2.07(m,1H),1.34(s,3H).LCMS(ESI,m/z):404.0[M+H]+.
第四步：向KH1-4(500mg,1.24mmol)的乙醇(10mL)溶液中分别加入Pd(dppf)Cl2(88mg,0.12mmol)和三乙胺(627mg,6.21mmoL)，一氧化碳氛围(四个标准大气压)下于120℃搅拌16h。反应完毕，减压蒸干溶剂，粗品经硅胶柱层析纯化得KH 1-5。1H NMR(400MHz,CDCl3)δ8.15(dd,J＝8.0,1.2Hz,1H),8.03(s,1H),7.93(d,J＝8.0Hz,1H),7.35(d,J＝8.6Hz,2H),6.84(d,J＝8.6Hz,2H),4.86(d,J＝15.4Hz,1H),4.48(d,J＝15.4Hz,1H),4.42(q,J＝7.2Hz,2H),3.78(s,3H),2.96(s,3H),2.71-2.59(m,2H),2.31-2.18(m,2H),1.43(t,J＝7.2Hz,3H),1.38(s,3H).LCMS(ESI,m/z):398.3[M+H]+.
第五步：KH 1-5(380mg,0.96mmol)的三氟乙酸(5mL)溶液在80℃下搅拌16h。反应完毕，减压蒸干溶剂，粗品经硅胶柱层析纯化得KH 1-6。1H NMR(400MHz,CDCl3)δ8.15(dd,J＝8.0,1.2Hz,1H),8.05(s,1H),7.88(d,J＝8.0Hz,1H),6.97(s,1H),4.42(q,J＝7.2Hz,2H),3.59-3.52(m,1H),3.49-3.41(m,1H),3.31(s,3H),2.16-2.09(m,1H),2.04-1.97(m,1H),1.56(s,3H),1.43(t,J＝7.2Hz,3H).LCMS(ESI,m/z):278.1[M+H]+.
第六步：向KH 1-6(200mg,0.72mmol)的DMF(5mL)溶液中加入氢化钠(58mg, 1.44mmol)，混合物室温搅拌30min，向混合物加入4-溴-2-氟苄溴(232mg,0.86mmol),混合物室温搅拌16h。反应完毕，加入水(10mL)，EA(10mL×3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥后，过滤，滤液浓缩的粗品经硅胶柱层析纯化得KH1-7。LCMS(ESI,m/z):464.0[M+H]+.
第七步：向KH 1-7(300mg,0.65mmol)的二氧六环(5mL)溶液中分别加入联硼酸频那醇酯(248mg,0.97mmol)，Pd(dppf)Cl2(44mg,0.06mmoL)和醋酸钾(128mg,1.31mmoL),混合物于80℃搅拌4h。反应完毕后，粗品不做处理直接用于下一步反应。LCMS(ESI,m/z):512.2[M+H]+.
第八步：向KH 1-8的反应体系分别加入水(0.5mL)，Int 1a(239mg,0.78mmol)，Pd(dppf)Cl2(44mg,0.06mmoL)和碳酸钾(181mg,1.31mmoL),混合物于90℃搅拌16h。反应完毕后，浓缩反应液，粗品经硅胶柱层析纯化得KH 1-9。LCMS(ESI,m/z):612.2[M+H]+.
第九步：向KH 1-9(150mg,0.24mmol)的甲醇(1.5mL)-水(0.5mL)混合溶液中加入氢氧化锂(20.16mg,0.48mmol)，混合物于60℃搅拌1h。反应完毕，减压蒸干溶剂，粗品经制备HPLC纯化得到KH 01。1H NMR(400MHz,DMSO)δ8.24(s,1H),8.07(d,J＝8.6Hz,1H),7.92-7.82(m,5H),7.78-7.69(m,2H),7.64(d,J＝7.4Hz,1H),7.48(t,J＝8.2Hz,1H),6.92(d,J＝8.2Hz,1H),5.60(s,2H),4.82(d,J＝16.2Hz,1H),4.67(d,J＝16.2Hz,1H),2.86(s,3H),2.65(d,J＝6.4Hz,2H),2.41-2.33(m,1H),2.27-2.22(m,1H),1.45(s,3H).LCMS(ESI,m/z):584.1[M+H]+.
第十步：化合物KH 01(27mg,0.04mmol)通过SFC手性拆分得KH01-1和KH01-2。其中KH01-1：LCMS(ESI,m/z):584.1[M+H]+.1H NMR(400MHz,DMSO)δ13.34(s,1H),8.23(s,1H),8.07(d,J＝8.0Hz,1H),7.94-7.81(m,5H),7.75-7.69(m,2H),7.64(d,J＝7.6Hz,1H),7.48(t,J＝8.2Hz,1H),6.91(d,J＝8.2Hz,1H),5.60(s,2H),4.81(d,J＝16.2Hz,1H),4.66(d,J＝16.2Hz,1H),2.86(s,3H),2.66(s,2H),2.41-2.32(m,1H),2.27-2.21(m,1H),1.45(s,3H).KH01-2：LCMS(ESI,m/z):584.1[M+H]+.1H NMR(400MHz,DMSO)δ13.34(s,1H),8.23(s,1H),8.07(d,J＝8.0Hz,1H),7.94-7.81(m,5H),7.75-7.69(m,2H),7.64(d,J＝7.6Hz,1H),7.48(t,J＝8.2Hz,1H),6.91(d,J＝8.2Hz,1H),5.60(s,2H),4.81(d,J＝16.2Hz,1H),4.66(d,J＝16.2Hz,1H),2.86(s,3H),2.66(s,2H),2.41-2.32(m,1H),2.27-2.21(m,1H),1.45(s,3H).
实施例2：2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH02)、(R)-2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6- 羧酸(KH02-1)、(S)-2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH02-2)的合成
第一步：向3-氟-4-甲酰基苯甲酸甲酯化合物KH2-1(5g,27.5mmol)的ACN(50mL)-H2O(10mL)混合溶液中，加入氨基磺酸(3.2g,32.9mmol)和次氯酸钠(4.9g,55.1mmol)，反应在室温下搅拌4h。反应完毕后，用1N盐酸溶液调pH至3-4。再加入乙酸乙酯(30mL×3)萃取，合并有机相，饱和食盐水洗涤，无水硫酸钠干燥，过滤，滤液浓缩得KH2-2。LCMS(ESI,m/z):199.2[M+H]+.
第二步：向KH 2-2(4.9g,24.7mmol)的DCM(50mL)溶液中，加入草酰氯(62.8g,494.8mmol)和DMF(20mg,0.27mmol)，混合物在室温搅拌16h，反应完毕，减压蒸干，粗品直接用于下一步。
第三步：向2-(4-溴-2,5-二氟苄基)肼-1-羧酸叔丁酯(Int 2a，7.5g，22.2mmol)的DCM(50mL)溶液中，加入中间体KH 2-3(4.1g,22.2mmol)和三乙胺(4.5g,44.4mmol)。反应在室温下搅拌2h。反应完毕，减压浓缩反应液，粗品经硅胶柱层析纯化得KH 2-4。LCMS(ESI,m/z):460.7[M+H-56]+.
第四步：向中间体KH 2-4(5g,9.6mmol)的乙酸乙酯(50mL)溶液中，加入4M盐酸的乙酸乙酯溶液(50mL)，混合物在室温下搅拌2h，反应完毕，浓缩得粗品KH2-5。LCMS(ESI,m/z):417.1[M+H]+.
第五步：将中间体KH 2-5(4.8g,11.51mmol)溶于DMSO(50mL)，将DIEA(4.46g,34.53mmol)加入反应体系，反应在150℃下搅拌2h。反应完成后，加入水(80mL)稀释反应体系，乙酸乙酯(30mL x 3)萃取，合并有机相，饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩反应液的粗品经硅胶柱层析纯化得KH 2-6。LCMS(ESI,m/z):398.8[M+H]+.1H NMR(400MHz,DMSO)δ7.86-7.74(m,3H),7.66(d,J＝8.2Hz,1H),7.35-7.26(m,1H),5.07(s,2H),3.89(s,3H).
第六步:将KH 2-6(0.5g,1.26mmol)溶于DMF(5mL)，依次将2-碘甲基氧杂环丁烷(0.32g,1.64mmol)和无水碳酸钾(0.35g,2.52mmol)加入反应体系，于50℃ 搅拌12h。反应完毕，加入水(30mL)稀释反应体系，乙酸乙酯(20mL×3)萃取，合并有机相，经饱和食盐水洗涤、无水硫酸钠干燥，过滤，滤液浓缩，粗品经硅胶柱层析纯化得KH 2-7。LCMS(ESI,m/z):468.9[M+H]+.1H NMR(400MHz,DMSO)δ8.10(s,1H),7.93(d,J＝8.8Hz,1H),7.79(s,1H),7.41(dd,J＝9.0,0.8Hz,1H),7.31(s,1H),5.56(s,2H),5.13-5.03(m,1H),4.73(dd,J＝11.8,4.0Hz,2H),4.57(d,J＝7.0Hz,2H),3.87(s,3H),2.79-2.57(m,2H).
第七步:将中间体KH 2-7(0.4g,0.86mmol)溶于1,4-二氧六环(5mL)，依次将联硼酸频哪醇酯(0.33g,1.28mmol)、KOAc(0.17g,1.71mmol)和Pd(dppf)Cl2(0.06g,0.09mmol)加入反应体系，于80℃下搅拌2h。反应完毕，蒸干溶剂，得到KH 2-8。LCMS(ESI,m/z):515.3[M+H]+.
第八步:将中间体KH 2-8(0.4g,0.77mmol)溶于1,4-二氧六环(5mL)-水(0.5mL)混合溶剂中，依次将Int 1a(0.24g,0.78mmol)、K2CO3(0.22g,1.55mmol)和Pd(dppf)Cl2(0.06g,0.08mmol)加入反应体系，反应在90℃下搅拌2h。反应完毕，减压浓缩，粗品经硅胶柱层析纯化得KH 2-9。LCMS(ESI,m/z):615.0[M+H]+.
第九步:中间体KH 2-9(0.11g)经SFC纯化得KH 2-9a(RT＝1.145min)和KH2-9b(RT＝1.529min)。
第十步：将中间体KH 2-9(30mg,0.048mmol)溶于甲醇(2.5mL)-水(0.5mL)混合溶液，将碳酸钾(14mg,0.099mmol)加入反应体系，50℃下搅拌3h。反应完毕，用水(10mL)稀释反应体系，调节pH值，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤合并的有机相，无水硫酸钠干燥，过滤，浓缩的粗品经制备HPLC纯化得KH 2。
采用相同水解条件，由KH 2-9a(40mg)和KH 2-9b(70mg)分别水解得到KH 02-1和KH02-2。其中，KH 02-1:1H NMR(400MHz,MeOD)δ8.10(s,1H),7.89-7.83(m,2H),7.77-7.72(m,1H),7.69(d,J＝7.8Hz,2H),7.62-7.53(m,2H),7.49(d,J＝8.0,1H),6.99(dd,J＝11.2,6.0Hz,1H),6.89(d,J＝8.2Hz,1H),5.59(s,2H),5.37(s,2H),4.80-4.76(m,1H),4.58-4.54(m,1H),4.48-4.36(m,2H),4.20(dd,J＝16.0,2.4Hz,1H),2.66-2.62(m,1H),2.49-2.38(m,1H).LCMS(ESI,m/z):601.1[M+H]+.KH02-2:LCMS(ESI,m/z):601.1[M+H]+.1H NMR(400MHz,MeOD)δ8.12(s,1H),7.89(d,J＝8.2Hz,1H),7.84(dd,J＝8.2,0.8Hz,1H),7.76(t,J＝8.0Hz,1H),7.71(dd,J＝6.0,2.0Hz,1H),7.69(d,J＝7.2Hz,1H),7.59(dd,J＝9.6,1.6Hz,1H),7.55(dd,J＝8.0,1.6Hz,1H),7.48(dd,J＝7.4,1.6Hz,1H),7.00(dd,J＝11.2,6.0Hz,1H),6.88(d,J＝8.4Hz,1H),5.58(s,2H),5.37(s,2H),4.80-4.76(m,1H),4.59-4.52(m,1H),4.48-4.35(m,2H),4.20(dd,J＝16.0,2.4Hz,1H),2.69-2.58(m,1H),2.51-2.38(m,1H).1H  NMR(400MHz,DMSO-d6)δ13.32(s,1H),8.14(s,1H),7.91(d,J＝9.6Hz,1H),7.86(t,J＝8.0Hz,1H),7.81(d,J＝8.0Hz,1H),7.76-7.67(m,4H),7.47(dd,J＝7.6,1.6Hz,1H),7.12(dd,J＝11.4,6.0Hz,1H),6.97(d,J＝8.4Hz,1H),5.56(s,2H),5.25(s,2H),4.69-4.60(m,1H),4.44-4.30(m,3H),4.20(dd,J＝16.2,2.6Hz,1H),2.61-2.53(m,1H),2.42-2.30(m,1H).
实施例3：2-(4-(6-((4-氯-2-氟苯并呋喃-7-基)甲氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH03)和(S)-2-(4-(6-((4-氯-2-氟苯并呋喃-7-基)甲氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH03-2)的合成
第一步:向中间体KH 2-8(77.1mg,0.15mmol)和Int 1b(53.40mg,0.15mmol)的二氧六环-水(Dioxane:H2O＝10:1，1mL)溶液中，加入Pd(dppf)Cl2(10.88mg,0.02mmol)和碳酸钾(41.46mg,0.30mmol)，氮气保护下于90℃搅拌3h。反应完毕后，加入10mL水，然后用乙酸乙酯萃取(10mL×3)，合并的有机相经饱和食盐水洗涤、无水硫酸钠干燥，过滤，浓缩的滤液经硅胶柱层析纯化得到KH 3-2。LCMS(ESI,m/z):664.1[M+H]+.
第二步：向中间体KH 3-2(40mg,0.06mmol)的甲醇-水(MeOH:H2O＝5:1，1mL)混合溶液中，加入碳酸钾(16.58mg,0.12mmol)，混合物在室温下搅拌1h。监测反应完成后，加水稀释，调节pH值，乙酸乙酯萃取。有机相经无水硫酸钠干燥后过滤，滤液减压蒸干，粗品经制备HPLC纯化得到KH 03。LCMS(ESI,m/z):650.3[M+H]+.1H NMR(400MHz,MeOD)δ8.05(s,1H),7.85(s,2H),7.73(t,J＝7.6Hz,1H),7.66(dd,J＝10.9,6.3Hz,1H),7.45(d,J＝6.8Hz,1H),7.37(d,J＝8.0Hz,1H),7.27(d,J＝8.2Hz,1H),6.98(dd,J＝11.4,5.9Hz,1H),6.85(d,J＝8.3Hz,1H),6.16(d,J＝6.5Hz,1H),5.67(s,2H),5.48(s,2H),5.38-5.36(m,2H),4.56-4.54(m,1H),4.40-4.33(m,1H),4.18-4.17(m,1H),2.63-2.61(m,1H),2.45-2.43(m,1H).
第三步：中间体KH 3-2(230mg,0.35mmol)经SFC纯化得KH 3-2a(RT＝1.706min)和KH 3-2b(RT＝2.813min)。
其中，中间体KH 3-2a。LCMS(ESI,m/z):664.1[M+H]+.1H NMR(400MHz,CDCl3)δ8.00-7.90(m,2H),7.83(d,J＝8.1Hz,1H),7.75(dd,J＝10.8,6.2Hz,1H),7.64(t,J＝7.8Hz,1H),7.45(d,J＝7.2Hz,1H),7.33(d,J＝8.1Hz,1H),7.24(s,1H),6.99(dd,J＝11.0,6.1Hz,1H),6.79(d,J＝8.1Hz,1H),6.02(d,J＝6.6Hz,1H),5.66(s,2H),5.34-5.25(m,2H),4.85-4.75(m,1H),4.63-4.53(m,1H),4.47-4.39(m,1H),4.19(dd,J＝16.1,6.9Hz,1H),4.03-3.96(m,1H),3.95(s,3H),2.65-2.53(m,1H),2.44-2.32(m,1H).
中间体KH 3-2b，LCMS(ESI,m/z):664.1[M+H]+.1H NMR(400MHz,CDCl3)δ7.99-7.91(m,2H),7.83(d,J＝8.1Hz,1H),7.75(dd,J＝10.9,6.2Hz,1H),7.64(t,J＝7.8Hz,1H),7.45(d,J＝6.7Hz,1H),7.33(d,J＝8.1Hz,1H),7.24(s,1H),6.99(dd,J＝10.9,5.9Hz,1H),6.79(d,J＝8.1Hz,1H),6.02(d,J＝6.6Hz,1H),5.66(s,2H),5.33-5.26(m,2H),4.80(d,J＝4.6Hz,1H),4.63-4.53(m,1H),4.47-4.40(m,1H),4.19(dd,J＝15.9,6.9Hz,1H),4.02-3.96(m,1H),3.95(s,3H),2.67-2.53(m,1H),2.44-2.32(m,1H).
拆分条件：Waters SFC 150；柱子：AS；尺寸：250*25mm10μm；流动相A：超临界CO2；流动相B：MeOH(+0.1％7.0mol/l甲醇氨溶液)；A:B：67:33；波长：214nm；流速：120ml/min；柱温:RT；背压:100bar；进样量:6.0mL；Cycle time:7.55min；溶剂:MeOH:蒸馏级；超临界CO2:食品级；样品溶液的制备:将样品溶解于约30mL MeOH)
第四步：向中间体KH 3-2b(110mg,0.16mmol)的THF(8mL)溶液中加入LiOH(7.9mg,0.33mmol)的水溶液(2mL)，室温搅拌1h。反应完毕，加入20mL水,用1M的盐酸调节pH＝3后,乙酸乙酯(30mL x 3)萃取，合并的有机相经饱和食盐水洗涤、无水硫酸钠干燥，过滤，滤液浓缩得到KH 03-2。LCMS(ESI,m/z):650.1[M+H]+.1H NMR(400MHz,CDCl3)δ8.03-7.97(m,2H),7.88(d,J＝8.4Hz,1H),7.76(dd,J＝10.8,6.0Hz,1H),7.64(t,J＝7.8Hz,1H),7.45(d,J＝6.8Hz,1H),7.33(d,J＝8.2Hz,1H),7.24(s,1H),7.01(dd,J＝11.0,5.7Hz,1H),6.79(d,J＝8.2Hz,1H),6.02(d,J＝6.6Hz,1H),5.66(s,2H),5.31(s,2H),4.87-4.76(m,1H),4.60(dd,J＝14.2,7.8Hz,1H),4.50-4.40(m,1H),4.23(dd,J＝16.0,7.4Hz,1H),4.07-3.93(m,1H),2.66-2.57(m,1H),2.45-2.36(m,1H).
实施例4：2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((1-(氰甲基)环丙基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH04)的合成
第一步:将中间体KH 2-6(1g,2.51mmol)溶于DMF(5mL)，依次将2-(1-(碘甲基)环丙基)乙腈(0.41g,3.77mmol)和无水碳酸钾(0.7g,5.06mmol)加入反应体系，反应在50℃下搅拌3h。反应完毕用水(30mL)稀释反应体系，乙酸乙酯(20mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得KH 4-2。LCMS(ESI,m/z):490.2[M+H]+.
第二步:将中间体KH 4-2(0.2g,0.40mmol)溶于1,4-二氧六环(3mL)，依次将联硼酸频那醇酯(0.16g,0.61mmol)、KOAc(0.08g,0.81mmol)和Pd(dppf)Cl2(0.03g,0.04mmol)加入反应体系，反应在80℃下搅拌2h。反应完毕，蒸干反应液，直接用于下一步。LCMS(ESI,m/z):538.2[M+H]+.
第三步:将中间体KH 4-3(0.2g,0.37mmol)溶于1,4-二氧六环(2mL)-水(0.2mL)混合溶剂中，依次将Int 1a(0.12g,0.37mmol)、K2CO3(0.10g,0.74mmol)和Pd(dppf)2Cl2(0.03g,0.04mmol)加入反应体系，于90℃下搅拌2h。反应完毕，减压蒸干溶剂，粗品经硅胶柱层析纯化得KH 4-4。LCMS(ESI,m/z):638.1[M+H]+.
第四步：将中间体KH 4-4(0.15g,0.23mmol)溶于甲醇(2.5mL)-水(0.5mL)混合溶剂中，再将K2CO3(0.06mg,0.47mmol)加入反应体系，室温下搅拌3h。反应完 毕，用水(10mL)稀释反应体系，调节(1M)PH至5-6，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩反应液，粗品经制备HPLC纯化得KH04。LCMS(ESI,m/z):624.2[M+H]+.1H NMR(400MHz,DMSO)δ13.34(s,1H),8.20(s,1H),7.92-7.82(m,3H),7.76-7.69(m,4H),7.46(d,J＝6.8Hz,1H),7.05-6.94(m,2H),5.56(s,2H),5.26(s,2H),4.04(s,2H),2.31(s,2H),0.71-0.65(m,2H),0.57-0.53(m,2H).
实施例5：2-(4-(6-((4-氯-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((1-(氰甲基)环丙基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH05)的合成
第一步:将中间体KH 4-3(0.1g,0.18mmol)溶于1,4-二氧六环(2mL)-水(0.2mL)混合溶液中，依次将2-溴-6-((4-氯-2-氟苄基)氧基)吡啶(0.06g,0.18mmol)、K2CO3(0.05g,0.36mmol)和Pd(dppf)Cl2(0.014g,0.019mmol)加入反应体系，反应在90℃下搅拌2h。反应完毕，浓缩反应液，粗品经硅胶柱层析纯化得KH 5-2。LCMS(ESI,m/z):647.2[M+H]+.
第二步：将中间体KH 5-2(0.13g,0.2mmol)溶于甲醇(2.5mL)和水(0.5mL)，LiOH(0.02mg,0.47mmol)加入反应体系，室温下搅拌3h。反应完毕，用水(10mL)稀释反应体系，盐酸(1M)调节PH至5-6，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩的滤液经制备HPLC纯化得KH05。LCMS(ESI,m/z):633.02[M+H]+.1H NMR(400MHz,DMSO)δ13.34(s,1H),8.20(s,1H),7.86-7.70(m,4H),7.57(t,J＝8.2Hz,1H),7.46(t,J＝9.2Hz,2H),7.30(d,J＝8.4Hz,1H),7.03(dd,J＝11.4,6.0Hz,1H),6.92(d,J＝8.4Hz,1H),5.46(s,2H),5.26(s,2H),4.04(s,2H),2.31(s,2H),0.70-0.65(m,2H),0.57-0.53(m,2H).
实施例6：2-(4-(6-((2-氟-4-氰基苄基)氧基)吡啶-2-基)-2-氟-5-甲基苄基)-1-((1-(氰甲基)环丙基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH06-2)的制备
第一步：中间体Int 2c(3.41g,10.24mmol)和三乙胺(2.59g,25.70mmol)溶于二氯甲烷(40mL)，0℃滴加中间体KH 2-3(2.79g,12.85mmol)的二氯甲烷溶液，加完后，保温反应2h。反应结束后，加水(30mL)淬灭，用二氯甲烷(3×150mL)萃取，盐水洗涤有机相，硫酸钠干燥后过滤，浓缩除去溶剂，残余物经硅胶柱层析纯化得KH 6-2。LCMS(ESI,m/z)：457[M+H-56]+。1H NMR(400MHz,CDCl3)δ7.77(d,J＝7.4Hz,1H),7.65(d,J＝9.8Hz,1H),7.41-7.57(m,1H),7.23(dd,J＝12.2,8.4Hz,2H),5.23(br s,1H),4.49(br s,1H),3.87(s,3H),2.30(s,3H),1.17(s,9H).
第二步：向氯化氢的乙酸乙酯溶液(2N,50mL)中加入中间体KH 6-2(4.6g,8.98mmol),在室温下搅拌12h。反应完毕，减压浓缩后得粗品KH 6-3。LCMS(ESI，m/z)：413.2[M+H]+。1H NMR(400MHz,DMSO)δ7.81(d,J＝7.9Hz,1H),7.70(d,J＝9.7Hz,1H),7.59-7.54(m,2H),7.33(d,J＝7.8Hz,1H),4.82(s,2H),4.16-4.08(m,2H),3.88(s,3H),2.33(s,3H).
第三步：向中间体KH 6-3(2.8g,6.80mmol)的DMSO(30mL)溶液中，加入DIEA(4.77g,34.57mmol)，150℃反应2h。反应完毕，加入水(200mL)，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩滤液，粗品经硅胶柱层析纯化得KH 6-4。LCMS(ESI，m/z)：393.2[M+H]+。1H NMR(400MHz,CDCl3)δ7.89(s,1H),7.86(d,J＝2.4Hz,1H),7.55(s,1H),7.30-7.20(m,3H),5.05(s,2H),3.93(s,3H),2.28(s,3H).
第四步：向中间体KH 6-4(400mg,1.02mmol)的DMF(5mL)溶液中，加入碳酸钾(289mg,2.04mmol)和2-(1-(碘甲基)环丙基)乙腈(338mg,1.53mmol)，室温反应4h。加入水(200mL)，乙酸乙酯萃取，饱和食盐水洗涤有机相，无水硫酸钠干燥，过滤，浓缩滤液，粗品经硅胶柱层析纯化，得到KH 6-5P1和KH 6-5P2。其中，KH 6-5P1，LCMS(ESI，m/z)：486.3[M+H]+.1H NMR(400MHz,DMSO)δ8.07(s,1H),7.90(d,J＝9.0Hz,1H),7.56(d,J＝9.4Hz,1H),7.38(d,J＝9.0Hz,1H),7.29(d,J＝8.0Hz,1H),5.53(s,2H),4.55(s,2H),3.86(s,3H),2.87(s,2H),2.27(s,3H),0.81(t,J＝5.4Hz,2H),0.73(t,J＝5.4Hz,2H)；KH 6-5P2，LCMS(ESI，m/z)：486.3[M+H]+.1H NMR(400MHz,DMSO)δ8.21(s,1H),7.84(s,1H),7.71(d,J＝8.2Hz,1H),7.50(d,J＝9.6Hz,1H),7.13(d,J＝8.0Hz,1H),5.16(s,2H),4.01(s,2H),3.90(s,3H),2.28(s,2H),2.20(s,3H),0.65(d,J＝4.6Hz,2H),0.53(t,J＝5.4Hz,2H)。
第五步：向KH 6-5P1(200mg,0.41mmol)的二氧六环(3mL)溶液中，加入醋酸钾(81mg,0.83mmol)、联硼酸频那醇酯(136mg,0.54mmol)和Pd(dppf)Cl2(30mg,0.04mmol),氮气氛围下，80℃下搅拌4h生成KH 6-6P1。采用同样方法获得KH 6-6P2。KH 6-6P1和KH 6-6P2均不做处理直接用于下一步。
第六步：向KH 6-6P1的体系中加入1,4-二氧六环(3mL)-水(0.3mL)混合溶液中，再加入化合物Int 1a(164mg,0.54mmol)和碳酸钾(114mg,0.83mmol)、Pd(dppf)Cl2(30mg,0.04mmol)，氮气保护下，于90℃搅拌4h。反应完毕，浓缩经硅胶柱层析色谱纯化得KH 6-7P1。LCMS(ESI,m/z):633.6[M+H]+.
采用类似的方法，以KH 6-6P2为原料，反应完毕后，反应液浓缩，经硅胶柱层析纯化(EA:PE＝1:1)，得KH 6-7P2。LCMS(ESI,m/z):633.6[M+H]+.
第七步：向KH 6-7P1(180mg,0.31mmol)的甲醇溶液(2mL)中加入碳酸钾(78mg,0.57mmol)和水(2mL)，50℃下搅拌2h。反应完毕，用1N盐酸调节pH至4， 再加入水20mL，乙酸乙酯萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥后，过滤，减压浓缩滤液，粗品经制备HPLC纯化，得KH06-1。LCMS(ESI,m/z):619.6[M+H]+.1H NMR(400MHz,DMSO)δ8.06(s,1H),7.88-7.82(m,3H),7.73-7.67(m,2H),7.38(d,J＝9.8Hz,1H),7.26-7.20(m,3H),6.94(d,J＝8.4Hz,1H),5.59(s,2H),5.49(s,2H),4.56(s,2H),2.87(s,2H),2.21(s,3H),0.83-0.80(m,2H),0.74-0.71(m,2H).
以KH 6-7P2为原料，采用相同的方法，得到KH06-2。LCMS(ESI,m/z):619.6[M+H]+.1H NMR(400MHz,DMSO)δ13.14(s,1H),8.18(s,1H),7.89-7.77(m,3H),7.71-7.68(m,3H),7.19-7.14(m,2H),7.05(d,J＝7.8Hz,1H),6.92(d,J＝8.4Hz,1H),5.47(s,2H),5.22(s,2H),4.04(s,2H),2.29(s,2H),2.14(s,3H),0.68-0.65(m,2H),0.55-0.53(m,2H).
实施例7：2-(4-(6-((1-(2,2-二氟乙基)-1H-吲唑-6-基)甲氧基)吡啶-2-基)-2-氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH07)的合成
第一步：向中间体Int 2c(2.6g,8.2mmol)的DCM(30mL)溶液中，加入三乙胺(2.8g,27.7mmol)和原料KH 2-3(3g,13.8mmol)，反应在室温下搅拌2h，反应完毕，减压浓缩反应液，粗品经硅胶柱层析纯化得KH 7-2。LCMS(ESI,m/z):443.0[M+H-56]+。
第二步：向中间体KH 7-2(1.8g,3.6mmol)的乙酸乙酯(10mL)溶液中，加入3M盐酸乙酸乙酯(10mL)，混合物在室温下搅拌12h，反应完毕，浓缩得粗品KH 7-3。LCMS(ESI,m/z):399.0[M+H]+。
第三步：将中间体KH 7-3(0.4g,1.0mmol)溶于DMSO(10mL)，将DIEA(0.38g,3.0mmol)加入反应体系，反应在150℃下搅拌2h，反应完毕，加入水(30mL)稀释反应体系，乙酸乙酯(20mL×3)萃取，饱和食盐水洗涤有机相，无水硫酸钠干燥，过滤，滤液浓缩，粗品经硅胶柱层析纯化得KH 7-4。LCMS(ESI,m/z):379.0[M+H]+.1H NMR(400MHz,DMSO)δ7.85-7.78(m,2H),7.67(m,1H),7.59(dd,J＝9.8,2.0Hz,1H),7.42(dd,J＝8.2,1.6Hz,1H),7.22(t,1H),5.07(s,2H),3.88(s,3H).
第四步：将中间体KH 7-4(0.1g,0.26mmol)溶于DMF(5mL)，依次将2-(碘甲基)环氧丁烷(0.67g,0.33mmol)和碳酸钾(0.73g,0.52mmol)加入反应体系，反应在50℃下搅拌12h。反应完毕，加入水(30mL)稀释反应体系，乙酸乙酯(20mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱层析纯化得KH 7-5。LCMS(ESI,m/z):449.2[M+H]+.
第五步：将中间体KH 7-5(0.05g,0.11mmol)溶于1,4-二氧六环(2mL)，依次将联硼酸频哪醇酯(0.04g,0.16mmol)、KOAc(0.02g,0.22mmol)和Pd(dppf)Cl2 (0.008g,0.01mmol)加入反应体系，反应在80℃下搅拌2h。反应完毕，浓缩，得KH 7-6。LCMS(ESI,m/z):497.3[M+H]+.
第六步：将中间体KH 7-6(0.05g,0.1mmol)溶于1,4-二氧六环(1mL)和水(0.1mL)，依次将中间体Int 1b(0.03g,0.1mmol)、K2CO3(0.02g,0.15mmol)和Pd(dppf)Cl2(6mg,0.008mmol)加入反应体系，反应在90℃下搅拌2h。反应完毕，浓缩，粗品经硅胶柱纯化得KH 7-7。LCMS(ESI,m/z):658.3[M+H]+.
第七步：将中间体KH 7-7(0.05g,0.07mmol)溶于甲醇(2.5mL)-水(0.5mL)混合溶液，将LiOH(3.6mg,0.15mmol)加入反应体系，室温下搅拌3h。反应完毕用水(10mL)稀释，1M盐酸调节PH至3-4，乙酸乙酯(10mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经制备HPLC纯化得KH07。LCMS(ESI,m/z):644.2[M+H]+.1H NMR(400MHz,MeOD)δ8.05(d,J＝0.7Hz,1H),7.93(s,1H),7.89-7.67(m,7H),7.44(d,J＝7.4Hz,1H),7.33(d,J＝9.3Hz,1H),7.17(t,J＝7.9Hz,1H),6.83(d,J＝8.1Hz,1H),6.19(tt,J＝55.2,3.8Hz,1H),5.62(s,2H),5.39(m,2H),4.82-4.73(m,3H),4.53(m,1H),4.43(m,1H),4.37-4.31(m,1H),4.18-4.12(m,1H),2.65-2.54(m,1H),2.49-2.36(m,1H)。
实施例8：2-(4-(6-(2-氟-4-氰基苄氧基)吡啶-2-基)-2-氟-5-甲基苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH08)的制备
第一步：向中间体KH 6-4(300mg,0.76mmol)的DMF(5mL)溶液中，加入2-(1-(碘甲基)环丙基)乙腈(166.68mg,0.84mmol)和碳酸钾(209.76mg,1.52mmol)，混合物50℃反应2h。反应完毕，加入水，乙酸乙酯萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥，过滤，减压浓缩，粗品经硅胶柱层析色谱纯化，得KH 8-2。LCMS(ESI，m/z)：463.2[M+H]+。
第二步：向中间体KH 8-2(200mg,0.43mmol)的二氧六环(2mL)溶液中，加入醋酸钾(85mg,0.87mmol)，联硼酸频那醇酯(143mg,0.56mmol)和Pd(dppf)Cl2(32mg,0.04mmol),氮气保护下，于80℃搅拌4h生成KH 8-3。不做处理直接用于下一步。LCMS(ESI，m/z)：510.3[M+H]+。
第三步：向中间体KH 8-3加入1,4-二氧六环(1mL)和水(0.1mL)，再加入化合物Int 1a(156mg,0.51mmol)和碳酸钾(108mg,0.78mmol)、Pd(dppf)Cl2(29mg,0.04mmol)，氮气氛围下，于90℃搅拌4h。反应完毕，减压浓缩，粗品经硅胶柱层析纯化得KH 8-4。LCMS(ESI,m/z):610.6[M+H]+.
第四步：向中间体KH 8-4(148mg,0.24mmol)的甲醇溶液(1mL)中加入碳酸钾(67mg,0.49mmol)和水(1mL)，50℃下搅拌2h。反应后，反应液加水稀释，调节pH值5-6，乙酸乙酯萃取，有机相经干燥后过滤，滤液浓缩，粗品经制备HPLC纯化 得KH08。LCMS(ESI,m/z):596.5[M+H]+.1H NMR(400MHz,DMSO)δ8.12(s,1H),7.90-7.78(m,3H),7.71-7.66(m,3H),7.21-7.18(m,2H),7.12(d,J＝7.6Hz,1H),6.93(d,J＝8.3Hz,1H),5.48(s,2H),5.20(s,2H),4.67-4.58(m,1H),4.35-4.22(m,4H),2.58-2.52(m,1H),2.38-2.33(m,1H),2.16(s,3H).
实施例9：2-(4-(6-((1-(2,2-二氟乙基)-1H-吲唑-6-基)甲氧基)吡啶-2-基)-2-氟-5-甲基苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH09)的合成
第一步：向中间体KH 8-3(130mg,0.25mmol)的1,4-二氧六环(2mL)-水(0.2mL)混合溶液中，加入中间体Int 1c(122mg,0.33mmol)和碳酸钾(71mg,0.51mmol)、Pd(dppf)Cl2(19mg,0.03mmol)，N2置换3次，90℃搅拌4h。反应完毕，减压浓缩后的粗品经硅胶柱层析纯化得KH 9-2。LCMS(ESI,m/z):671.6[M+H]+.
第二步：向中间体KH 9-2(130mg,0.19mmol)的甲醇溶液(2mL)中加入碳酸钾(66mg,0.48mmol)和水(2mL)，50℃下搅拌2h。反应后，反应液加水稀释，调节pH值5-6，乙酸乙酯萃取，有机相经干燥后过滤，滤液浓缩，粗品经制备HPLC纯化得KH09。LCMS(ESI,m/z):657.6[M+H]+.1H NMR(400MHz,DMSO)δ8.12(d,J＝17.1Hz,2H),7.81-7.76(m,4H),7.67(d,J＝8.1Hz,1H),7.27-7.22(m,2H),7.16-7.12(m,2H),6.91(d,J＝8.3Hz,1H),6.39(t,J＝55.0Hz,1H),5.48(s,2H),5.20(s,2H),4.93-4.83(m,2H),4.64-4.62(m,1H),4.41-4.16(m,4H),2.54-2.52(m,1H),2.36-2.32(m,1H),2.20(s,3H).
实施例10：2-(4-(6-((1-(2,2-二氟乙基)-1H-吲唑-6-基)甲氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH10)的合成
第一步:向中间体KH 2-8(90mg,0.18mmol)和中间体Int 1c(64.40mg,0.18mmol)的二氧六环-水(Dioxane:H2O＝10:1，1mL)溶液中，加入Pd(dppf)Cl2(1.38mg,0.02mmol)和碳酸钾(48.37mg,0.35mmol)，氮气氛围下90℃搅拌3h。反应完 毕，减压浓缩反应液，粗品经硅胶柱层析纯化得到KH 10-2。LCMS(ESI,m/z):676.4[M+H]+.
第二步：向中间体KH 10-2(100mg,0.15mmol)的甲醇-水(MeOH:H2O＝5:1，1mL)溶液中，加入碳酸钾(40.89mg,0.30mmol)，混合物在50℃下搅拌1h。1N盐酸调节PH至3，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经制备HPLC纯化得到KH 10。LCMS(ESI,m/z):662.1[M+H]+.1H NMR(400MHz,DMSO)δ8.15(s,1H),8.02(br s,1H),7.83-7.80(m,4H),7.69-7.67(m,2H),7.45(d,J＝6.4Hz,1H),7.30(d,J＝8.0Hz,1H),7.11-7.03(m,1H),6.95(d,J＝8.4Hz,1H),6.54-6.26(m,1H),5.57(s,2H),5.22(s,2H),4.89-4.87(m,2H),4.64-4.62(m,1H),4.36-4.35(m,3H),4.17-4.16(m,1H),2.54-2.52(m,1H),2.39-2.35(m,1H).
实施例11：2-(4-(6-(2-氟-4-氰基苄氧基)吡啶-2-基)-2-氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH11)的制备
第一步:将中间体KH 7-6(0.1g,0.2mmol)溶于1,4-二氧六环(2mL)和水(0.2mL)的混合溶液中，依次将中间体Int 1a(0.12g,0.4mmol)、K2CO3(0.05g,0.4mmol)和Pd(dppf)Cl2(0.015g,0.02mmol)加入反应体系，反应在90℃下搅拌2h。反应完毕，浓缩反应液，粗品经硅胶柱层析纯化得KH 11-2。LCMS(ESI,m/z):597.1[M+H]+.
第二步：将中间体KH 11-2(0.1g,0.17mmol)溶于乙腈(2mL)，将TBD(0.9M,0.4mL)加入反应体系，室温下搅拌2h。反应完毕用水(5mL)稀释，1N盐酸调节PH至3，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤，滤液浓缩，粗品经制备HPLC纯化得KH11。LCMS(ESI,m/z):583.2[M+H]+.1H NMR(400MHz,DMSO)δ8.06(s,1H),7.91(d,J＝10.1Hz,1H),7.86-7.80(m,3H),7.78-7.66(m,4H),7.61(d,J＝7.5Hz,1H),7.17(t,J＝8.2Hz,1H),6.91(d,J＝8.2Hz,1H),5.58(s,2H),5.24(s,2H),4.67-4.58(m,1H),4.42-4.27(m,3H),4.22-4.13(m,1H),2.54(s,1H),2.39-2.31(m,1H).
实施例12：2-(4-(6-(2-氟-4-氰基苄氧基)-5-氟吡啶-2-基)-2-氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH12)的制备
第一步:将中间体KH 7-6(0.1g,0.2mmol)溶于1,4-二氧六环(2mL)和水(0.2mL)混合溶液，依次将中间体Int 1d(0.065g,0.2mmol)、K2CO3(0.05g,0.4mmol)和Pd(dppf)Cl2(0.015g,0.02mmol)加入反应体系，反应在90℃下搅拌2h。反应完毕，浓缩反应液，粗品经硅胶柱层析纯化得KH 12-2。LCMS(ESI,m/z):615.1[M+H]+。
第二步：将中间体KH 12-2(0.1g,0.16mmol)溶于乙腈(2mL)，将TBD(0.9M,0.4mL)加入反应体系，室温下搅拌2h。反应完毕用水(5mL)稀释，1N盐酸调节PH至3，乙酸乙酯(5mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤，滤液浓缩，粗品经制备HPLC纯化得KH12。LCMS(ESI,m/z):601.1[M+H]+.1H NMR(400 MHz,DMSO)δ8.12(s,1H),7.95-7.90(m,1H),7.84-7.63(m,8H),7.20(t,J＝8.0Hz,1H),5.67(s,2H),5.25(s,2H),4.69-4.58(m,1H),4.43-4.27(m,3H),4.22-4.15(m,1H),2.58-2.52(m,1H),2.40-2.28(m,1H)。
实施例13：2-(4-(6-(2-甲氧基-4-氰基苄氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH13)的制备
第一步:。向中间体KH 2-8(329.17mg,0.64mmol)和中间体Int 1e(204.26mg,0.64mmol)的二氧六环和水(Dioxane:H2O＝10:1，3mL)溶液中，加入Pd(dppf)Cl2(46.44mg,0.06mmol)和碳酸钾(176.91mg,1.28mmol)，氮气氛围下90℃搅拌3h。反应完毕，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，粗品经硅胶柱纯化得到KH 13-2。LCMS(ESI,m/z):627.6[M+H]+.
第二步：向中间体KH 13-2(310mg,0.50mmol)的乙腈(3mL)溶液中，加入TBD的水溶液(0.9M,1.2mL)，混合物在室温下搅拌2h。浓缩，得粗品经Prep-HPLC纯化得到KH13。LCMS(ESI,m/z):613.1[M+H]+.1H NMR(400MHz,DMSO)δ8.12(s,1H),7.90-7.77(m,2H),7.69-7.63(m,2H),7.55(d,J＝7.8Hz,2H),7.43(dd,J＝12.6,4.8Hz,2H),7.11(dd,J＝11.3,6.0Hz,1H),6.97(d,J＝8.2Hz,1H),5.47(s,2H),5.24(s,2H),4.66-4.62(m,1H),4.44-4.32(m,3H),4.21-4.16(m,1H),3.89(s,3H),2.70-2.52(m,1H),2.42-2.31(m,1H).
实施例14：2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2-氟苄基)-1-((1-(氟甲基)环丙基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH14)的合成
第一步：向KH 7-4(920mg,2.427mmol)和中间体Int 1f(1.191g,3.641mmol)的DMF(10mL)溶液中,加入K2CO3(670mg,4.855mmol),50℃搅拌2h。反应完毕，加入水(30mL)，用乙酸乙酯(30mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得到KH 14-2。LCMS(ESI,m/z):577.1[M+H]+.1H NMR(400MHz,CDCl3)δ8.01-7.92(m,2H),7.82(dd,J＝8.2,1.2Hz,1H),7.26(dd,J＝9.5,1.9Hz,1H),7.17(dd,J＝8.3,1.5Hz,1H),6.96(t,J＝8.1Hz,1H),5.31(s,2H),3.98(s,3H),3.92(s,2H),3.12(s,2H),0.89(s,9H),0.53-0.47(m,2H),0.47-0.41(m,2H),0.11(s,3H),0.00(s,3H).
第二步：向中间体KH 14-2(520mg,0.901mmol)的THF(10mL)溶液中加入TBAF(1.35mL,1M)的THF溶液，室温搅拌2h。反应完毕，加入水(30mL)，用乙酸乙酯(30mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得到KH 14-3。LCMS(ESI,m/z):463.1[M+H]+.1H NMR(400MHz,CDCl3)δ7.96(s, 1H),7.85(d,J＝8.2Hz,1H),7.73(dd,J＝8.2,1.2Hz,1H),7.17-7.13(m,1H),7.08(dd,J＝8.3,1.5Hz,1H),6.91(t,J＝8.2Hz,1H),5.23(s,1H),5.20(s,2H),3.88(s,3H),3.81(s,2H),3.05-3.01(m,2H),0.48-0.44(m,2H),0.43-0.39(m,2H).
第三步：向中间体KH 14-3(210mg,0.454mmol)的二氯甲烷(8mL)溶液中加入DAST(146mg,0.907mmol)，-78℃搅拌1h。反应完毕,用饱和NaHCO3水溶液淬灭反应，二氯甲烷(30mL×3)萃取，硫酸钠干燥，浓缩，粗产品经硅胶柱纯化得到KH14-4。LCMS(ESI,m/z):465.1[M+H]+.1H NMR(400MHz,CDCl3)δ7.94-7.91(m,2H),7.83-7.79(m,1H),7.25-7.22(m,1H),7.16(dd,J＝8.4,1.6Hz,1H),7.02(t,J＝8.1Hz,1H),5.24-5.19(m,2H),4.10(s,1H),4.04(s,1H),3.95(s,4H),3.91-3.85(m,1H),2.28-2.12(m,2H),2.05-1.93(m,2H).
第四步：向中间体KH 14-4(120mg,0.258mmol)和联硼酸频哪醇酯(98mg,0.387mmol的二氧六环(10mL)溶液中加入Pd(dppf)Cl2(28mg,0.0387mmol)，KOAc(51mg,0.516mmol),氮气氛围80℃搅拌6h。反应完毕,所得的KH 14-5直接用于下一步。LCMS(ESI,m/z):513.2[M+H]+.
第五步：向含有中间体KH 14-5(11.5mL)的反应液中加Int 1a(71mg,0.232mmol),Pd(dppf)Cl2(28mg,0.0387mmol)，碳酸钾(71mg,0.516mmol),水(1mL),氮气氛围90℃搅拌12h。反应完毕,加水(30mL)，乙酸乙酯(40mL×3)萃取，硫酸钠干燥，浓缩，粗产品经硅胶柱纯化后得到KH 14-6。LCMS(ESI,m/z):613.2[M+H]+.1H NMR(400MHz,CDCl3)δ8.00-7.90(m,2H),7.85-7.78(m,1H),7.72-7.57(m,4H),7.47-7.37(m,2H),7.34-7.28(m,1H),7.20(br,1H),6.79(d,J＝7.8Hz,1H),5.58(s,2H),5.36-5.27(m,3H),4.19-4.06(m,2H),4.00-3.84(m,4H),2.31-2.13(m,2H),1.71-1.56(m,2H).
第六步：向中间体KH 14-6(110mg,0.179mmol)的乙腈(8mL)溶液中加入TBD(0.45mL,0.9M in H2O),室温搅拌2h。反应完毕，加水(20mL)，用HCl(1M)调节pH＝3，乙酸乙酯(30mL×3)萃取，饱和食盐水洗涤，硫酸钠干燥，浓缩，粗品经Prep-HPLC纯化后得到KH14。LCMS(ESI,m/z):599.2[M+H]+.1H NMR(400MHz,DMSO)δ13.25(s,1H),8.18(s,1H),7.90(d,J＝9.7Hz,1H),7.87-7.78(m,4H),7.77-7.65(m,3H),7.60(d,J＝7.5Hz,1H),7.06(t,J＝7.9Hz,1H),6.90(d,J＝8.2Hz,1H),5.58(s,2H),5.26(s,2H),4.41(d,J＝23.6Hz,2H),2.12-1.94(m,4H),1.81-1.58(m,2H).
实施例15：(R)-2-(4-(6-((2-氟-4-氯苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH15-1)、(S)-2-(4-(6-((2-氟-4-氯苄基) 氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH15-2)的制备
第一步：向中间体KH 2-8(329.17mg,0.64mmol)和2-溴-6-((4-氯-2-氟苄基)氧基)吡啶(202.88mg,0.64mmol)的二氧六环和水(Dioxane:H2O＝10:1，3mL)溶液中，加入Pd(dppf)Cl2(46.44mg,0.06mmol)和碳酸钾(176.91mg,1.28mmol)，氮气氛围下90℃搅拌3h。反应完毕，加入水(10mL)，乙酸乙酯(10mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩。经硅胶柱纯化得到KH 15-2a。
第二步：中间体KH 15-2a经SFC分离得到KH 15-3a(保留时间1.288min)和KH15-3b(保留时间2.09min)。
第三步：向中间体KH 15-3a(160mg,0.26mmol)的甲醇和水(MeOH:H2O＝5:1，2mL)溶液中，加入氢氧化锂(12.30mg,0.51mmol)，混合物在室温下搅拌1h。反应完毕，浓缩，得粗品经Prep-HPLC纯化得到KH15-1。LCMS(ESI,m/z):610.1[M+H]+.1H NMR(400MHz,DMSO)δ13.29(s,1H),8.13(s,1H),7.87-7.76(m,3H),7.67-7.72(m,1H),7.58(t,J＝8.2Hz,1H),7.45-7.50(m,2H),7.28-7.33(m,1H),7.12(dd,J＝11.3,5.9Hz,1H),6.92(d,J＝8.2Hz,1H),5.46(s,2H),5.25(s,2H),4.61-4.69(m,1H),4.45-4.30(m,3H),4.25-4.16(m,1H),2.67-2.54(m,1H),2.42-2.31(m,1H).
第四步：向中间体KH 15-3b(180mg,0.28mmol)的甲醇和水(MeOH:H2O＝5:1，2mL)溶液中，加入氢氧化锂(15.38mg,0.57mmol)，混合物在室温下搅拌1h。反应完毕，浓缩，得粗品经Prep-HPLC纯化得到KH15-2。LCMS(ESI,m/z):610.1[M+H]+.1H NMR(400MHz,DMSO)δ13.29(s,1H),8.13(s,1H),7.87-7.76(m,3H),7.67-7.72(m,1H),7.58(t,J＝8.2Hz,1H),7.45-7.50(m,2H),7.29-7.33(m,1H),7.12(dd,J＝11.3,5.9Hz,1H),6.92(d,J＝8.2Hz,1H),5.46(s,2H),5.25(s,2H),4.63-4.68(m,1H),4.45-4.30(m,3H),4.25-4.16(m,1H),2.67-2.54(m,1H),2.42-2.31(m,1H).
实施例16：2-(4-(6-(2-氟-4-氰基苄氧基)吡啶-2-基)-3-氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH16)的制备
第一步：在0℃下向中间体Int 2e(1.45g,4.545mmol)和TEA(1.02g,10.10mmol)的DCM(20mL)溶液中，滴入中间体KH 2-3的DCM(20mL)溶液，加完后室温搅拌2h。反应完毕，加入水(30mL)，用DCM(40mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，经硅胶柱纯化得到KH 16-2。LCMS(ESI,m/z):499.1[M+H]+.1H NMR(400MHz,DMSO)δ9.93(s,1H),7.85-7.68(m,3H),7.60-7.49(m,1H),7.41-7.32(m,1H),7.18(d,J＝8.2Hz,1H),5.09(d,J＝14.4Hz,1H),4.44(d,J＝16.2Hz,1H), 3.92-3.87(m,3H),1.17(s,9H).
第二步：向中间体KH 16-2(2.0g,4.008mmol)的甲醇(10mL)溶液中，加入盐酸乙酸乙酯(25mL,3M)溶液，混合物室温搅拌2h。反应完毕，浓缩，加入水(30mL)，饱和NaHCO3水溶液调节pH＝8,DCM(50mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩后得到KH 16-3。LCMS(ESI,m/z):399.0[M+H]+.1H NMR(400MHz,DMSO)δ7.81(dd,J＝8.0,1.6Hz,1H),7.77-7.68(m,2H),7.59-7.53(m,1H),7.30(dd,J＝9.8,2.0Hz,1H),7.14(dd,J＝8.2,1.8Hz,1H),4.82(s,2H),4.81(s,2H),3.88(s,3H).
第三步：向中间体KH 16-3(1.5g,3.759mmol)的DMSO(15mL)溶液中，加入DIEA(1.0g,7.769mmol)，150℃下搅拌3h。反应完毕，加入水(40mL)，用EA(50mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经正相柱纯化得到KH 16-4。LCMS(ESI,m/z):379.0[M+H]+.1H NMR(400MHz,CDCl3)δ7.90-7.79(m,3H),7.51-7.45(m,1H),7.09-7.04(m,1H),7.00-6.95(m,1H),4.95(s,2H),3.88(s,3H).
第四步：向中间体KH 16-4(520mg,1.425mmol)和K2CO3(393mg,2.850mmol)的DMF(10mL)溶液中，加入2-碘甲烷氧杂环丁烷(367mg,1.852mmol)，50℃下搅拌16h。反应完毕，加入水(30mL)，用EA(40mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经过正相柱纯化得到KH 16-5。LCMS(ESI,m/z):449.0[M+H]+.1H NMR(400MHz,CDCl3)δ7.94(d,J＝8.2Hz,1H),7.87(s,1H),7.84(d,J＝8.0Hz,1H),7.50-7.43(m,1H),7.01(dd,J＝9.0,1.6Hz,1H),6.95-6.88(m,1H),5.25(d,J＝16.4Hz,1H),5.13(d,J＝16.4Hz,1H),4.85-4.75(m,1H),4.59(dd,J＝14.0,8.0Hz,1H),4.46-4.35(m,1H),4.05(dd,J＝16.2,6.6Hz,1H),3.95(s,3H),3.89(dd,J＝16.2,2.8Hz,1H),2.66-2.52(m,1H),2.43-2.30(m,1H).
第五步：向中间体KH 16-5(320mg,0.713mmol)和联硼酸频哪醇酯(272mg,1.069mmol)的1，4-二氧六环(15mL)溶液中，加入Pd(dppf)Cl2(78mg,0.107mmol)和KOAc(140，mg,1.425mmol),氮气氛围下于80℃搅拌12h。反应完毕，KH 16-6反应液直接用于下一步。LCMS(ESI,m/z):497.2[M+H]+.
第六步：向中间体KH 16-6(15mL 0.713mmol)的反应液中加入中间体Int 1a(186mg,0.606mmol),Pd(dppf)Cl2(78mg,0.107mmol),K2CO3(197mg,1.425mmol)和H2O(1.5mL)，氮气氛围90℃搅拌2h。反应完毕，浓缩，粗品经硅胶柱层析色谱纯化得到KH 16-7。LCMS(ESI,m/z):597.2[M+H]+.1H NMR(400MHz,CDCl3)δ7.90(d,J＝8.2Hz,1H),7.86-7.80(m,2H),7.77(d,J＝8.2Hz,1H),7.62-7.53(m,2H),7.37(d,J＝7.6Hz,2H),7.31(d,J＝9.4Hz,1H),7.04(d,J＝8.0Hz,1H),6.94(d,J＝12.6Hz,1H),6.72(d,J＝8.2Hz,1H),5.49(s,2H),5.31-5.12(m,2H),4.82-4.69(m,1H),4.53 (dd,J＝14.2,8.0Hz,1H),4.40-4.31(m,1H),4.03(dd,J＝16.0,6.4Hz,1H),3.91-3.83(m,4H),2.57-2.45(m,1H),2.36-2.23(m,1H).
第七步：向中间体KH 16-7(260mg,0.436mmol)的THF(10mL)溶液中加入LiOH(21mg,0.871mmol)的H2O(2mL)溶液，室温下搅拌1h。反应完毕，加入水(20mL)，用1M的盐酸调节pH＝3，EA(40mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩后经过制备HPLC纯化得到KH16。LCMS(ESI,m/z):583.2[M+H]+.1H NMR(400MHz,DMSO)δ8.14(s,1H),7.95-7.79(m,4H),7.76-7.66(m,3H),7.46-7.39(m,1H),7.24-7.12(m,2H),6.93(d,J＝8.2Hz,1H),5.54(s,2H),5.24(s,2H),4.70-4.61(m,1H),4.48-4.32(m,3H),4.27-4.19(m,1H),2.60-2.52(m,1H),2.41-2.31(m,1H).
实施例17：2-(4-(6-((4-(环丙基羰基)-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH17)的制备
第一步:向原料KH 17-1(1g,5.43mmol)和2-氟-6-溴吡啶(956.5mg，5.43mmol)的二氧六环(10mL)溶液中，加入2mL叔丁醇钾(730mg，6.53mmol)的四氢呋喃溶液，室温搅拌，逐渐升温到40℃反应0.5h。反应完毕，加入水(10mL)和乙酸乙酯(10mL x 3)萃取，有机相经饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH 17-2。LCMS(ESI,m/z):341.9[M+H]+.1H NMR(400MHz,DMSO)δ7.84-7.81(m,1H),7.76-7.68(m,3H),7.28(d,J＝7.6Hz,1H),6.97(d,J＝8.2Hz,1H),5.44(s,2H),3.88(s,3H).
第二步:向中间体KH 17-2(890m g,2.62mmol)的甲醇(10mL)溶液中，加入溶于水(2mL)的氢氧化锂(125mg，5.35mmol)溶液，室温搅拌反应2h。反应完毕，用1N盐酸调节pH＝3,加入水(10mL)和乙酸乙酯(10mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥、过滤，滤液浓缩得到KH 17-3。LCMS(ESI,m/z):341.9[M+H]+.1H NMR(400MHz,DMSO)δ13.31(s,1H),7.80(d,J＝8.0Hz,1H),7.75-7.64(m,3H),7.28(d,J＝7.4Hz,1H),6.97(d,J＝8.2Hz,1H),5.43(s,2H).
第三步:向中间体KH 17-3(750mg,2.3mmol)和甲氧基甲基胺盐酸盐(337mg，3.45mmol)的二氯甲烷(10mL)溶液中，加入TCFH(840mg，3.0mmol)与NMI(566mg，6.9mmol)，室温搅拌1h。反应完毕，加入水(10mL)和乙酸乙酯(10mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥后过滤，滤液浓缩后经硅胶柱层析色谱纯化得到KH 17-4。LCMS(ESI,m/z):370.9[M+H]+.1H NMR(400MHz,DMSO)δ7.70(t,J＝7.8Hz,1H),7.64(t,J＝7.8Hz,1H),7.49-7.43(m,2H),7.28(d,J＝7.6Hz,1H),6.96(d,J＝8.2Hz,1H),5.40(s,2H),3.57(s,3H),3.27(s,3H).
第四步:向中间体KH 17-4(780mg,2.12mmol)的超干四氢呋喃(10mL)溶液中，缓慢加入环丙基溴化镁(1.54g，10.6mmol)，0℃搅拌3h。然后，加入饱和氯化铵水溶液(10mL)，用乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,过滤，滤液浓缩，粗品经硅胶柱层析色谱纯化得到KH 17-5。LCMS(ESI,m/z):351.9[M+H]+.
第五步:向中间体KH 17-5(150mg,0.43mmol)的二氧六环(3mL)与水(0.3mL)混合溶液中，加入中间体KH 2-8(200g，0.43mmol)、碳酸钾(118.7mg,0.56mmol)与Pd(dppf)Cl2(31.4mg,0.043mmol)，90℃搅拌2.5h。反应结束，浓缩反应液，粗品经硅胶柱层析色谱纯化得到KH 17-6。LCMS(ESI,m/z):658.2[M+H]+.
第六步:向中间体KH 17-6(220mg,0.334mmol)的乙腈(2mL)溶液中，加入TBD的水溶液(0.9mol，0.8mL)，室温搅拌2.5h。反应完毕，加入水(5mL)，用乙酸乙酯(5mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，浓缩所得的粗品经制备HPLC纯化得到KH17。LCMS(ESI,m/z):644.1[M+H]+.1H NMR(400MHz,DMSO)δ13.33(s,1H),8.13(s,1H),7.89-7.84(m,3H),7.80(d,J＝8.2Hz,1H),7.76-7.73(m,1H),7.71-7.68(m,2H),7.47(dd,1H),7.12(dd,J＝11.2,6.2Hz,1H),6.97(d,J＝8.2Hz,1H),5.57(s,2H),5.24(s,2H),4.64(m,1H),4.43-4.29(m,3H),4.20(d,J＝13.2Hz,1H),2.93-2.89(m,1H),2.35-2.29(m,2H),1.07-1.01(m,4H).
实施例18：2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2-氟苄基)-4-氟-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH18)的制备
第一步：向原料KH 18-1(10g,42.3mmol)和TEA(21.398g,211.9mmol)的甲醇(100mL)溶液中，加入Pd(dppf)Cl2(2.34g,4.23mmol)，一氧化碳加压氛围下120℃搅拌12h。反应完毕，过滤浓缩，得粗品经Prep-HPLC纯化得到KH 18-2。LCMS(ESI,m/z):214.8[M-H]-.H NMR(400MHz,DMSO)δ7.77-7.60(m,2H),3.90(s,3H).
第二步：向中间体KH 18-2(8g,37.0mmol)二氯甲烷(80mL)溶液中，加入草酰氯(94.1g,740.7mmol)与催化量DMF，室温搅拌12h。反应完毕，浓缩，得粗品KH 18-3直接进行下一步反应。
第三步：向中间体Int 2b(8.7g,27.4mmol)和TEA(5.52g,54.7mmol)的二氯甲烷(60mL)溶液中，滴加中间体KH 18-3，室温搅拌2.5h。反应完毕，加入水(100mL)，二氯甲烷(100mL x 3)萃取，饱和食盐水洗涤有机相，无水硫酸钠干燥,过滤，浓缩，粗品经硅胶柱层析色谱纯化得KH 18-4。LCMS(ESI,m/z):580.0[M+Na+ACN]+.
第四步：向中间体KH 18-4(13.7g,26.5mmol)的乙酸乙酯(100mL)溶液中，加入氯化氢乙酸乙酯溶液(100mL)，室温搅拌12h。反应完毕，过滤得到KH 18-5。LCMS(ESI,m/z):419.0[M+H]+.1H NMR(400MHz,DMSO)δ7.64(d,J＝7.4Hz,2H),7.59(dd,J＝9.6,1.8Hz,1H),7.49(dd,J＝8.2,1.6Hz,1H),7.33-7.28(m,1H),4.93(s,2H),4.86(s,2H),3.89(s,3H).
第五步：向中间体KH 18-5(1g,2.4mmol)的二甲基亚砜(10mL)溶液中，加入DIEA(928mg,7.2mmol)，150℃搅拌2h。反应完毕，加入水(20mL)，乙酸乙酯(20mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩得粗品，经制备HPLC纯化,得到KH 18-6。LCMS(ESI,m/z):397.0[M+H]+.1H NMR(400MHz,DMSO)δ11.13(s,1H),7.62-7.58(m,2H),7.43(dd,J＝8.2,1.6Hz,1H),7.25(d,J＝2.4Hz,1H),7.23-7.21(m,1H),5.06(s,2H),3.88(s,3H).
第六步：向中间体KH 18-6(470mg,1.18mmol)和2-碘甲基氧杂环丁烷(257.2mg,1.3mmol)的二甲基亚砜(5mL)溶液中，加入DIEA(304.6mg,2.36mmol)，50℃搅拌12h。反应完毕，水(10mL)和乙酸乙酯(10mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩，粗品经硅胶柱层析色谱纯化得到KH 18-7。LCMS(ESI,m/z):469.0[M+H]+.1H NMR(400MHz,DMSO)δ8.01(d,J＝0.8Hz,1H),7.56(dd,J＝9.8,2.0Hz,1H),7.38(dd,J＝8.4,1.8Hz,1H),7.29(d,J＝9.8Hz,1H),7.13(t,J＝8.2Hz,1H),5.24-5.11(m,2H),4.70-4.62(m,1H),4.45-4.39(m,2H),4.36-4.30(m,1H),4.20(dd,J＝16.2,2.4Hz,1H),3.89(s,3H),2.58-2.48(m,1H),2.38-2.28(m,1H).
第七步：向中间体KH 18-7(158mg,0.34mmol)和Int 1a(129mg,0.51mmol)的四氢呋喃(3mL)溶液中，加入乙酸钾(66.3mg,0.68mmol)和Pd(dppf)Cl2(24.9mg,0.034mmol)，80℃搅拌2.5h。反应完毕，所得粗品KH 18-8直接用于下一步。LCMS(ESI,m/z):515.2[M+H]+.
第八步：向上一步的反应液中，加入水(0.03mL)、碳酸钾(93.4mg,0.68mmol)和Pd(dppf)Cl2(24.9mg,0.034mmol)，90℃搅拌2.5h。反应完毕，加入水(10mL)和乙酸乙酯(10mL×3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥,过滤，浓缩，粗品经硅胶柱层析色谱纯化得到KH 18-9。LCMS(ESI,m/z):615.1[M+H]+.
第九步：向中间体KH 18-9(120mg,0.195mmol)的乙腈(2mL)溶液中，加入TBD的水溶液(0.9mol，0.4ml)，室温搅拌2.5h。反应完毕，加入水(5mL)和乙酸乙酯(5mL×3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥,过滤，浓缩，所得粗品经制备HPLC纯化得到KH18。LCMS(ESI,m/z):601.2[M+H]+.1H NMR(400MHz,DMSO)δ13.55(s,1H),7.96(s,1H),7.91(d,J＝9.8Hz,1H),7.86(d,1H),7.83(d,2H),7.77-7.69(m,2H),7.62(d,J＝7.6Hz,1H),7.29-7.20(m,2H),6.91(d,J＝8.2Hz,1H),5.59(s,2H),5.24(s,2H),4.68-4.66(m,1H),4.39-4.36(m,3H),4.22-4.20(m,1H),2.34-2.32(m,2H).
实施例19：2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2,6-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH 19)的制备
第一步：向中间体KH 2-3(1g，4.62mmol)的二氯甲烷(10mL)溶液中加入中间体Int 2d(1.56mg，4.62mmol)和三乙胺(1.2mL)，于室温搅拌2h。反应完全后，真空减压浓缩得粗品经硅胶柱层析色谱纯化得KH 19-2。LCMS(ESI，m/z)：416.9[M+1]+。1H NMR(400MHz,DMSO)δ7.82-7.69(m,2H),7.55-7.43(m,3H),5.10-4.56(m,2H),3.86(s,3H),1.16(s,9H).
第二步：向中间体KH 19-2(2g，3.87mmol)的乙酸乙酯(20mL)溶液中加入氯化氢乙酸乙酯(20mL)，于室温下反应2h。反应完全后，将反应液过滤，并用乙酸乙酯冲洗滤饼，将滤饼干燥得到KH 19-3。LCMS(ESI，m/z)：419.0[M+1]+。1H NMR(400MHz,DMSO)δ7.78(s,1H),7.65(s,1H),7.54-7.40(m,3H),4.86(s,2H),3.86(d,J＝4.2Hz,3H).
第三步：向中间体KH 19-3(950mg，2.28mmol)的二甲基亚砜(10mL)溶液中加入DIEA(1.2mL)，于150℃下反应2h。反应完全后，将反应液倒入水中，加入乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，真空减压浓缩得粗品；再将粗品用硅胶柱层析纯化后得KH 19-4。LCMS(ESI，m/z)：397.0[M+1]+。1H NMR(400MHz,DMSO)δ10.34(s,1H),7.80-7.74(m,2H),7.68-7.64(m,1H),7.53(d,J＝7.2Hz,2H),5.05(s,2H),3.87(s,3H).
第四步：向KH 19-4(100mg，0.25mmol)的DMF(2mL)溶液中加入2-碘甲基 氧杂环丁烷(65mg，0.33mmol)和碳酸钾(70mg，0.50mmol)，于50℃下反应4h。反应完全后，将反应液倒入水中，加入乙酸乙酯萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥，过滤，真空减压浓缩得粗品，粗品经硅胶柱层析色谱纯化得KH 19-5。LCMS(ESI，m/z)：469.0[M+1]+。1H NMR(400MHz,DMSO)δ8.08(s,1H),7.96(s,1H),7.78(d,J＝8.2Hz,1H),7.72-7.65(m,1H),7.42(d,J＝7.6Hz,1H),5.18(s,2H),4.65-4.55(m,1H),4.43-4.31(m,2H),4.28-4.20(m,1H),4.15-4.06(m,1H),3.89(s,3H),2.58-2.52(m,1H),2.37-2.29(m,1H).
第五步：向中间体KH 19-5(110mg，0.24mmol)的1，4-二氧六环(3mL)溶液中加入联硼酸频哪醇酯(90mg，0.35mmol)，乙酸钾(46mg，0.47mmol)和Pd(dppf)Cl2(17mg，0.024mmol)，于80℃在氮气保护下反应2h。反应完全后，产物KH 19-6直接用于下一步反应。LCMS(ESI，m/z)：433.0[M+1]+。
第六步：向中间体KH19-6(110mg，0.25mmol)的1，4-二氧六环(2mL)和水(0.5mL)的混合溶液中加入中间体Int 1a(78mg，0.25mmol)，碳酸钾(70mg，0.51mmol)和Pd(dppf)Cl2(18mg，0.025mmol)，于90℃在氮气保护下反应12h。反应完全后，浓缩，粗品用硅胶柱层析纯化后得KH 19-7。LCMS(ESI，m/z)：615.2[M+1]+。1H NMR(400MHz,DMSO)δ8.11(s,1H),7.94-7.67(m,10H),5.60(d,J＝10.2Hz,2H),5.32-5.20(m,2H),4.58(d,J＝8.4Hz,1H),4.42-4.26(m,3H),4.15(d,J＝14.2Hz,1H),3.87(s,3H),2.54(s,1H),2.33(s,1H).
第七步：向中间体KH 19-7(47mg，0.076mmol)的乙腈(2mL)溶液中加入0.9M TBD(0.2mL)的水溶液，于室温搅拌2h。反应完全后，加稀盐酸调pH＝5，加入乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，真空减压浓缩后粗品用反相制备色谱纯化得KH19。LCMS(ESI，m/z)：601.1[M+1]+。1H NMR(400MHz,DMSO)δ8.02(s,1H),7.91(d,J＝9.8Hz,1H),7.88-7.84(m,1H),7.80-7.64(m,8H),6.95(d,J＝8.2Hz,1H),5.59(s,2H),5.24(q,J＝16.0Hz,2H),4.56(d,J＝6.3Hz,1H),4.42-4.33(m,2H),4.27-4.20(m,1H),4.11(d,J＝14.2Hz,1H),2.52(s,1H),2.33(s,1H).
实施例20：2-(4-(6-((5-氯吡啶-2-基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH20)的制备
第一步：向原料KH 20-1(1g,7.0mmol)和2-氟-6-溴吡啶(1.2g,7.0mmol)的二氧六环(10mL)溶液中，加入t-BuOK(940mg,8.4mmol)，混合物在40℃下搅拌0.5h。反应完毕，加入水(10mL)，用乙酸乙酯(10mL x 3)萃取，合并的有机相经饱和食盐水洗涤、无水硫酸钠干燥,过滤，滤液浓缩所得粗品经硅胶柱层析色谱纯化得到KH20-2。LCMS(ESI,m/z):300.9[M+H]+.1H NMR(400MHz,DMSO)δ8.62(d,J＝2.4Hz, 1H),7.97(dd,J＝8.4,2.4Hz,1H),7.71(t,1H),7.53(d,J＝8.4Hz,1H),7.27(d,J＝7.6Hz,1H),6.99(d,J＝8.2Hz,1H),5.39(s,2H).
第二步：向中间体KH 20-2(45mg,0.15mmol)与中间体KH 2-8(77mg,0.15mmol)的二氧六环(2ml)溶液中，加入水(0.02mL)，碳酸钾(93.4mg,0.68mmol)和Pd(dppf)Cl2(24.9mg,0.034mmol)，90℃搅拌2.5h。反应完毕，加入水(5mL)，乙酸乙酯(5mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥,过滤，滤液浓缩所得粗品经硅胶柱层析色谱纯化得到KH 20-3。LCMS(ESI,m/z):607.1[M+H]+.
第三步：向中间体KH 20-3(40mg,0.065mmol)的乙腈(2mL)溶液中，加入的TBD水溶液(0.9mol，0.2ml)，混合物室温搅拌2.5h。反应完毕，用1N盐酸调节pH＝3，加入水(5mL)，乙酸乙酯(5mL x 3)萃取，合并有机相并用饱和食盐水洗涤，无水硫酸钠干燥有机相后,过滤，浓缩滤液，所得粗品经制备HPLC纯化得到KH 20。LCMS(ESI,m/z):593.0[M+H]+.1H NMR(400MHz,DMSO)δ8.61(d,J＝2.4Hz,1H),8.12(s,1H),7.93(dd,J＝8.6,2.4Hz,1H),7.86(t,1H),7.80(d,J＝8.2Hz,1H),7.68(d,J＝8.0Hz,1H),7.63(dd,J＝10.8,6.2Hz,1H),7.51(d,J＝8.4Hz,1H),7.47(d,1H),7.10(dd,J＝11.4,5.6Hz,1H),6.99(d,J＝8.2Hz,1H),5.50(s,2H),5.23(s,2H),4.65-4.61(m,1H),4.35-4.31(m,3H),4.19(d,J＝14.2Hz,1H),2.55-2.51(m,1H),2.41-2.34(m,1H).
实施例21：(S)-2-(4-(6-(2-氘代甲氧基-4-氰基苄氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH21-2)的制备
第一步:将原料KH 21-1(2g,7.19mmol)溶于DMF(20mL)，将氰化亚铜(1.5g,16.85mmol)加入反应体系，反应在140℃下搅拌2h。反应完毕，冷却至室温加入水(80mL)淬灭，乙酸乙酯(30mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得KH 21-2。1H NMR(400MHz,DMSO)δ10.83(s,1H),7.91(d,J＝8.2Hz,1H),7.49(d,J＝12Hz,1H),7.40(dd,J＝8.1,1.4Hz,1H),3.94(s,3H).
第二步:将中间体KH 21-2(1.2g,6.77mmol)溶于乙腈(10mL)，将氘代碘甲烷(1.9g,13.10mmol)、碳酸铯(4.4g,13.50mmol)加入反应体系，反应室温搅拌6h。反应完毕，浓缩，粗品经硅胶柱纯化得KH 21-3。1H NMR(400MHz,DMSO)δ7.76(d,J＝7.8Hz,1H),7.67(d,J＝1.2Hz,1H),7.49(dd,J＝7.8,1.3Hz,1H),3.83(s,3H).
第三步:将中间体KH 21-3(1g,5.15mmol)溶于THF(10mL)，将LiBH4(5mL,2.0M)、MeOH(0.33g,10.31mmol)加入反应体系，反应50℃搅拌2h。反应完毕，冷却至室温，加入氯化铵饱和溶液(50mL)淬灭，乙酸乙酯(30mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得KH 21-4。1H NMR(400MHz,DMSO)δ7.55(d,J＝7.7Hz,1H),7.43(d,J＝7.8Hz,1H),7.39(s,1H),5.29(t,J＝5.6Hz,1H),4.53(d,J＝5.6Hz,2H).
第四步:将中间体KH 21-4(0.2g,1.19mmol)溶于1,4-二氧六环(3mL)，将2-溴-6-氟吡啶(0.21g，1.19mmol)和叔丁醇钾(1.4mL,1M)加入反应体系，反应在45℃ 下搅拌30min。反应完毕，冷却至室温，浓缩，粗品经硅胶柱纯化得KH 21-5。LCMS(ESI,m/z):322.0[M+H].
第五步:将中间体KH 21-5(0.22g,0.67mmol)溶于1,4-二氧六环(3mL)和水(0.3mL)，将中间体KH 2-8(0.3g，0.64mmol)、碳酸钾(0.17g,1.28mmol)和Pd(dppf)Cl2(0.05g，0.06mmol)加入反应体系，反应90℃搅拌2h。反应完毕，浓缩，粗品经硅胶柱纯化得KH 21-6。LCMS(ESI,m/z):630.2[M+H].
第六步：中间体KH 21-6(0.2g)经SFC[型号：C2AS，条件：1525％MeOH DEA C2AS]分离得到KH 21-6a和KH 21-6b，保留时间分别为1.597分钟和2.553分钟。
第七步：将中间体KH 21-6b(0.1g,0.15mmol)溶于乙腈(2mL)，将TBD(0.9M,0.4mL)加入反应体系，室温下搅拌2h。反应完毕用水(5mL)稀释，稀醋酸调节PH至5-6，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经Prep-HPLC纯化得KH21-2。LCMS(ESI,m/z):616.2[M+H]+.1H NMR(400MHz,DMSO)δ8.13(s,1H),7.87-7.79(m,2H),7.72-7.64(m,2H),7.55(s,1H),7.53(t,J＝5.2Hz,1H),7.47-7.40(m,2H),7.11(dd,J＝11.4,6.0Hz,1H),6.97(d,J＝8.2Hz,1H),5.47(s,2H),5.24(s,2H),4.71-4.60(m,1H),4.43-4.32(m,3H),4.20(dd,J＝16.0,2.4Hz,1H),2.59-2.53(m,1H),2.40-2.31(m,1H).
实施例22：2-(4-(6-((2-氟-4-氯苯并呋喃-7-基)甲氧基)吡啶-2-基)-2,6-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH22)的制备
第一步:向中间体KH 19-6(143.92mg,0.28mmol)和中间体Int 1b(100mg,0.28mmol)的二氧六环和水(Dioxane:H2O＝10:1，2mL)溶液中，加入Pd(dppf)Cl2(21.77mg,0.03mmol)和碳酸钾(77.40mg,0.56mmol)，氮气氛围下90℃搅拌2h。反应完毕，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓 缩。经硅胶柱纯化得到KH 22-2。LCMS(ESI,m/z):664.0[M+H]+.
第二步：向中间体KH 22-2(130mg,0.20mmol)的甲醇和水(MeOH:H2O＝5:1，2mL)溶液中，加入氢氧化锂(9.6mg,0.40mmol)，混合物在室温下搅拌1h。浓缩，得粗品经制备HPLC纯化得到KH22。LCMS(ESI,m/z):650.0[M+H]+.1H NMR(400MHz,DMSO)δ13.26(s,1H),8.07(s,1H),7.87-7.81(m,1H),7.73-7.78(m,3H),7.70-7.64(m,2H),7.41(dd,J＝19.9,8.2Hz,2H),6.93(d,J＝8.2Hz,1H),6.49(d,J＝6.4Hz,1H),5.68(s,2H),5.19-5.32(m,2H),4.53-4.63(m,1H),4.45-4.32(m,2H),4.22-4.29(m,1H),4.09-4.15(m,1H),2.51-2.56(m,1H),2.27-2.37(m,1H).
实施例23-27：2-(4-(6-((2-甲氧基-4-氯苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH23)、2-(4-(6-((2,6-二氟-4-氯苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH24)、2-(4-(6-((2,4-二氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH25)、2-(4-(6-((2-氯-4-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH26)、(S)-2-(4-(6-((4-氯-2-(二甲氨酰基)苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸的制备(KH27)。
采用实施例3相同的合成方法，分别以中间体Int 1f-1～Int 1f-4为原料与中间体KH 2-8(或S构型的中间体KH 2-8b)反应，分别得到目标分子KH23～KH27。
表3



实施例28：(S)-2-(4-(6-((4-氰基-呋喃并[3,2-c]吡啶-7-基)甲氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH28)的制备
第一步：将原料KH 28-1(10g,73.74mmol)溶于乙腈(100mL)，0℃滴加NBS(14.6g,73.74mmol)并搅拌1h。反应完毕，浓缩，粗品经甲醇打浆得KH 28-2。LCMS(ESI,m/z):213.9[M+H]+.1H NMR(400MHz,DMSO)δ8.00(d,J＝2.1Hz,1H),7.60(s,1H),7.06(d,J＝2.1Hz,1H).
第二步:将中间体KH 28-2(3g,14.01mmol)溶于乙醇(30mL)，将三乙胺(7g,69.17mmol)、Pd(dppf)Cl2(1g,1.36mmol)加入反应体系，反应于一氧化碳环境下100℃搅拌16h。反应完毕，过滤浓缩，粗品经甲基叔丁基醚打浆得KH 28-3。LCMS(ESI,m/z):208.1[M+H].
第三步:将KH 28-3(2g,9.61mmol)溶于三氯氧磷(5mL)，反应120℃回流搅拌1h。反应完毕，冷却至室温，浓缩，粗品缓慢加入冰水浴(80mL)淬灭，乙酸乙酯(30mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得KH 28-4。LCMS(ESI,m/z):226.0[M+H].1H NMR(400MHz,CDCl3)δ8.87(s,1H),7.83(d,J＝2.0Hz,1H),6.96(d,J＝2.2Hz,1H),4.50(q,J＝7.1Hz,2H),1.46(t,J＝7.1Hz,3H).
第四步:将中间体KH 28-4(0.5g,2.21mmol)溶于THF(10mL)，将LiBH4(2.6mL,2.0M)、MeOH(0.17g,5.31mmol)加入反应体系，反应50℃搅拌1h。反应完毕，冷却至室温，加入氯化铵饱和溶液(30mL)淬灭，乙酸乙酯(20mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得KH 28-5。LCMS(ESI,m/z):184.8[M+H].1H NMR(400MHz,DMSO)δ8.30-8.19(m,2H),7.12(d,J＝2.4Hz,1H),5.51(t,J＝5.6Hz,1H),4.81(d,J＝5.5Hz,2H).
第五步:将中间体KH 28-5(0.23g,1.25mmol)溶于DMF(3mL)，将氰化锌(0.29g，2.50mmol)和Pd(dppf)Cl2(0.09mg,0.12mmol)加入反应体系，反应在120℃下搅拌3h。反应完毕，冷却至室温，加入水(20mL)稀释，乙酸乙酯(10mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得KH 28-6。LCMS(ESI,m/z):175.1[M+H].
第六步:将中间体KH 28-6(0.18g,1.02mmol)溶于1,4-二氧六环(2mL)，将2-氟-6-溴吡啶(0.18g，1.02mmol)和叔丁醇钾(1mL,1M)加入反应体系，反应在45℃下搅拌30min。反应完毕，冷却至室温，浓缩，粗品经硅胶柱纯得KH 28-7。LCMS(ESI,m/z):330.0[M+H].1H NMR(400MHz,CDCl3)δ8.74(s,1H),7.88(d,J＝2.2Hz,1H),7.47(t,J＝7.8Hz,1H),7.15-7.04(s,2H),6.78(d,J＝8.1Hz,1H),5.76(s,2H).
第七步：将中间体KH 28-7(0.13g,0.27mmol)溶于1,4-二氧六环(2mL)和水(0.2mL)，将中间体KH 2-8b(0.1g，0.27mmol)、碳酸钾(0.07g,0.55mmol)和Pd(dppf)Cl2(0.02g，0.02mmol)加入反应体系，反应90℃搅拌2h。反应完毕，浓缩， 粗品经硅胶柱纯化得KH 28-8。LCMS(ESI,m/z):638.0[M+H].
第八步：将中间体KH 28-8(0.08g,0.13mmol)溶于乙腈(2mL)，将TBD(0.9M,0.2mL)加入反应体系，室温下搅拌2h。反应完毕用水(5mL)稀释，盐酸(1M)调节PH至5-6，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经Prep-HPLC纯化得KH28。LCMS(ESI,m/z):624.1[M+H]+.1H NMR(400MHz,DMSO)δ8.77(s,1H),8.42(d,J＝2.0Hz,1H),8.13(s,1H),7.85(d,J＝7.8Hz,1H),7.81(d,J＝8.2Hz,1H),7.69(d,J＝8.9Hz,2H),7.47(d,J＝7.0Hz,1H),7.39(d,J＝2.0Hz,1H),7.11(dd,J＝11.2,5.9Hz,1H),6.98(d,J＝8.2Hz,1H),5.85(s,2H),5.24(s,2H),4.69-4.61(m,1H),4.44-4.32(m,3H),4.20(d,J＝14.6Hz,1H),2.56-2.53(m,1H),2.41-2.31(m,1H).
实施例29、实施例30：(S)-2-(4-(4-((4-氰基-2-甲氧基苄基)氧基)嘧啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH29)和(S)-2-(4-(2-((4-氰基-2-甲氧基苄基)氧基)嘧啶-4-基)-2,5-二氟苄基)-1-((氧杂环丁烷-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH30)的制备
第一步:向原料KH 29-1(5g,31.03mmol)的甲醇和四氢呋喃(MeOH:THF＝1:1，50mL)溶液中，加入NaBH4(1.17g,31.03mmol)，室温搅拌1h。反应完毕，浓缩，所得粗品KH 29-2直接用于下一步。1H NMR(400MHz,DMSO)δ7.53-7.69(m,1H),7.47-7.33(m,2H),5.32(t,J＝5.6Hz,1H),4.53(d,J＝5.5Hz,2H),3.84(s,3H).
第二步:向中间体KH 29-2(1g,6.13mmol)的DMF(10mL)溶液中，加入碳酸铯(4.00g,12.26mmol)和2,4-二溴嘧啶(1.46g,6.13mmol)，室温下搅拌1h。反应完毕，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，有机相经饱和食盐水洗涤、无水硫酸钠干燥，过滤，滤液浓缩后，经硅胶柱层析色谱纯化得到KH 29-3a和KH 29-3b混合物。其中，350mg混合物经SFC分离得到KH 29-3a(Rt＝1.796min)和KH 29-3b(Rt＝2.627min)。
KH 29-3a谱图数据：LCMS(ESI,m/z):320.0[M+H]+.1H NMR(400MHz,DMSO)δ8.40-8.47(m,1H),7.57-7.50(m,2H),7.40-749(m,1H),7.08(d,J＝5.7Hz,1H),5.39(s,2H),3.86(s,3H)；
KH 29-3b谱图数据：1H NMR(400MHz,DMSO)δ8.50(d,J＝5.2Hz,1H),7.54-7.57(m,2H),7.51(d,J＝5.2Hz,1H),7.44-7.48(m,1H),5.43(s,2H),3.90(s,3H).
第三步:向中间体KH 29-3a(107.94mg,0.21mmol)和KH 2-8b(67.2mg,0.21mmol)的二氧六环和水(Dioxane:H2O＝10:1，2mL)溶液中，加入Pd(dtbp)Cl2(13.05mg,0.02mmol)和碳酸钾(58.05mg,0.42mmol)，氮气氛围下60℃搅拌3h。反应完毕，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH 29-4。LCMS(ESI,m/z):628.2[M+H]+.
第四步：向KH 29-4(90mg,0.14mmol)的乙腈(1mL)溶液中，加入TBD(0.9M,0.72mL)，混合物在室温下搅拌1h。浓缩，得粗品经Prep-HPLC纯化得到KH 29。LCMS(ESI,m/z):614.2[M+H]+.1H NMR(400MHz,DMSO)δ13.32(s,1H),8.66-8.75(m,1H),8.13(s,1H),7.87-7.77(m,2H),7.65-7.74(m,1H),7.59-7.53(m,2H),7.42-7.47(m,1H),7.22-7.09(m,1H),7.04(d,J＝5.8Hz,1H),5.49-5.56(m,2H),5.26(s,2H),4.61-4.69(m,1H),4.44-4.30(m,3H),4.17-4.24(m,1H),3.86-3.92(m,3H),2.50-2.59(m,1H),2.40-2.30(m,1H).
第五步:向中间体KH 29-3b(67.2mg，0.21mmol)和KH 2-8b(107.94mg,0.21mmol)的二氧六环和水(Dioxane:H2O＝10:1，2mL)溶液中，加入Pd(dppf)Cl2(14.51mg,0.02mmol)和碳酸钾(58.05mg,0.42mmol)，氮气氛围下90℃搅拌3h。反应完毕，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH 30-1。LCMS(ESI,m/z):628.1[M+H]+.
第六步：向中间体KH 30-1(90mg,0.14mmol)的乙腈(1mL)溶液中，加入TBD(0.9M,0.72mL)，混合物在室温下搅拌1h。浓缩，得粗品经Prep-HPLC纯化得到KH30。LCMS(ESI,m/z):614.1[M+H]+.1H NMR(400MHz,DMSO)δ8.74(d,J＝5.2Hz,1H),8.13(s,1H),7.86-7.79(m,2H),7.67-7.72(m,1H),7.58-7.53(m,3H),7.43-7.47(m,1H),7.18(dd,J＝11.3,5.9Hz,1H),5.50(s,2H),5.27(s,2H),4.62-4.67(m,1H),4.33-4.43(m,3H),4.17-4.22(m,1H),3.90(s,3H),2.58-2.52(m,1H),2.33-2.39(m,1H).
实施例31：(S)-2-(4-(6-((4-甲氧基-2-氰基吡啶-5-基)甲氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH31)的制备
第一步:向原料KH 31-1(5g,24.5mmol)和甲醇钠(1.97g,36.4mmol)的甲醇(100mL)溶液中，室温搅拌12h。反应完毕，加入水(50mL)，乙酸乙酯(50mL x 3) 萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH 31-2。LCMS(ESI,m/z):201.9[M+H]+.1H NMR(400MHz,DMSO)δ8.56(s,1H),7.36(s,1H),3.96(s,3H),3.82(s,3H).
第二步:向中间体KH 31-2(1.5g,7.43mmol)四氢呋喃(15mL)溶液中，加入硼氢化锂四氢呋喃溶液(7.5ml,14.9mmol,2mol/L)，甲醇(0.63ml,14.9mmol)，50℃搅拌1h。反应完毕，滴加饱和氯化铵水溶液淬灭，加入水(15mL)，乙酸乙酯(15mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH 31-3。LCMS(ESI,m/z):173.9[M+H]+.1H NMR(400MHz,DMSO)δ8.17(s,1H),7.12(s,1H),5.22(t,J＝5.6Hz,1H),4.46(d,J＝5.0Hz,2H),3.89(s,3H).
第三步:向中间体KH 31-3(800mg,4.6mmol)和氰化锌(1.076g,9.2mmol)与Pd(dppf)Cl2(336mg,0.46mmol)的DMF(8mL)溶液中，120℃搅拌3h。反应完毕，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH 31-4。LCMS(ESI,m/z):165.2[M+H]+.1H NMR(400MHz,DMSO)δ8.53(s,1H),7.73(d,J＝1.7Hz,1H),5.44-5.38(m,1H),4.59-4.53(m,2H),3.93(s,3H).
第四步:向中间体KH 31-4(300mg,1.83mmol)和2-氟-6-溴吡啶(325mg,1.83mmol)的二氧六环(6mL)溶液中，滴加叔丁醇钾的四氢呋喃溶液(2.0mL,1mol/L)， 45℃搅拌0.5h。反应完毕，加入水(5mL)，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，得到粗品KH 31-5直接用于下一步。LCMS(ESI,m/z):319.9[M+H]+.
第五步:向中间体Int 31-5(360mg,1.12mmol)和KH 2-8b(574mg,1.12mmol)的二氧六环(6mL)与水(0.6ml)溶液中，加入碳酸钾(309mg,2.24mmol)与Pd(dppf)Cl2(81.9mg,0.112mmol)，70℃搅拌2.5h。反应完毕，加入水(5mL)，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH 31-6。LCMS(ESI,m/z):428.1[M+H]+.
第六步:向中间体KH 31-6(100mg,0.16mmol)的乙腈(2mL)溶液中，加入的氢氧化锂水溶液(0.3mol，7ml)，100℃搅拌5min。反应完毕，加入水(5mL)，滴加乙酸水溶液使溶液呈酸性，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，得粗品经Prep-HPLC纯化得到KH 31。LCMS(ESI,m/z):601.2[M+H]+.1H NMR(400MHz,DMSO)δ8.62(s,1H),8.13(s,1H),7.85-7.70(m,5H),7.46(d,J＝6.4Hz,1H),7.12(dd,J＝11.2,6.4Hz,1H),6.95(d,J＝8.2Hz,1H),5.47(s,2H),5.25(s,2H),4.65(dd,J＝13.6,2.4Hz,1H),4.43-4.31(m,3H),4.21-4.18(m,1H),3.97(s,3H),2.58-2.53(m,1H),2.38-2.33(m,1H).
实施例32：(S)-2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH32)的制备
第一步:向原料KH 32-1(1g,4.55mmol)和TEA(2.3g,22.73mmol)的甲醇(100mL)溶液中，加入Pd(dppf)Cl2(333mg,0.455mmol)，一氧化碳加压氛围下，于120℃搅拌12h。反应完毕，加入2.5％碳酸氢钠水溶液，乙酸乙酯萃取，水相用1N盐酸调节PH至5-6，再加入乙酸乙酯萃取，有机相经饱和食盐水洗涤，无水硫酸钠干燥，过滤，滤液浓缩得粗品为KH 32-2。LCMS(ESI,m/z):197.8[M-H]+.1H NMR(400MHz,DMSO)δ8.57(dd,J＝9.2,7.6Hz,1H),8.09(dd,J＝7.6,1.4Hz,1H),3.91(s,3H).
第二步:向中间体KH 32-2(800mg,4.02mmol)二氯甲烷(8mL)溶液中，加入草酰氯(10.21g,80.4mmol)与催化量DMF，室温搅拌12h。反应完毕，浓缩，得粗品KH 32-3直接进行下一步反应。
第三步:向中间体Int 2c(1.354g,4.02mmol)和TEA(812mg,8.04mmol)的二氯甲烷(8mL)溶液中，滴加中间体KH 32-3，室温搅拌3h。反应完毕，加入水(10mL)和二氯甲烷(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH 32-4。LCMS(ESI,m/z):519.9[M+H]+.1H NMR(400MHz,DMSO)δ8.08(m,2H),7.76(m,1H),7.50-7.42(m,1H),5.75(s,1H),5.05-4.57(m,2H),3.89(s,3H),1.16-1.11(m,9H).
第四步:向中间体KH 32-4(1.3g,2.17mmol)的乙酸乙酯(13mL)溶液中，加入盐酸乙酸乙酯溶液(13mL,4mol/L)，室温搅拌12h。反应完毕，过滤得到KH 32-5。LCMS(ESI,m/z):399.9[M+H]+.1H NMR(400MHz,DMSO)δ8.35(d,J＝8.0Hz,1H),7.82-7.73(m,2H),7.37(dd,J＝8.8,6.3Hz,1H),5.12(s,2H),3.91(s,3H).
第五步:向中间体KH 32-5(300mg,0.75mmol)和中间体Int 3(163mg,0.83mmol)的二甲基亚砜(3mL)溶液中，加入DIEA(193.5mg,1.5mmol)，100℃搅拌12h。反应完毕，加入水(5mL)，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，经硅胶柱纯化得到KH 32-6。LCMS(ESI,m/z):469.9[M+H]+.1H NMR(400MHz,DMSO)δ8.38(d,J＝7.9Hz,1H),7.83(d,J＝7.9Hz,1H),7.74(dd,J＝9.2,5.7Hz,1H),7.30(dd,J＝8.8,6.3Hz,1H),5.228-5.254(m,2H),4.774-4.750(m, 1H),4.418-4.288(s,3H),4.16(dd,J＝15.6,3.1Hz,1H),3.92(s,3H),2.63-2.54(m,1H),2.43-2.34(m,1H).
第六步:向中间体KH 32-6(160mg,0.34mmol)和硼烷频哪醇酯(130mg,0.51mmol)的二氧六环(2mL)溶液中，加入乙酸钾(67mg,0.68mmol)与Pd(dppf)Cl2(24.9mg,0.034mmol)，80℃搅拌2.5h。反应完毕，直接用于下一步反应。LCMS(ESI,m/z):516.1[M+H]+.
第七步:向中间体KH 32-7反应溶液中，加入中间体Int 1a(85mg,0.34mmol)、水(0.02mL)、碳酸钾(86mg,0.62mmol)与Pd(dppf)Cl2(23mg,0.031mmol)，90℃搅拌2.5h。反应完毕，浓缩，粗品经硅胶柱纯化(PE:EA＝2:1)得到KH 23-8。LCMS(ESI,m/z):616.0[M+H]+.
第八步:向中间体KH 32-8(80mg,0.13mmol)的乙腈(2mL)溶液中，加入的TBD水溶液(0.9mol，0.4ml)，室温搅拌1h。反应完毕，加入水(5mL)，滴加乙酸水溶液使溶液调成酸性，乙酸乙酯(5mL×3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，得粗品经Prep-HPLC纯化得到KH32。LCMS(ESI,m/z):602.0[M+H]+.1H NMR(400MHz,DMSO)δ8.20(d,J＝7.6Hz,1H),7.94-7.84(m,2H),7.76-7.70(m,3H),7.67(d,J＝8.0Hz,1H),7.47(d,J＝6.4Hz,1H),7.13(dd,J＝11.2,5.6Hz,1H),6.96(d,J＝7.6Hz,1H),5.56(s,2H),5.27(s,2H),4.74(m,1H),4.45-4.29(m,3H),4.28-4.21(m,1H),2.68-2.54(m,1H),2.38-2.31(m,1H).
实施例33：(S)-2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-4-氟-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH33)的制备
第一步:向起始物料KH33-1(20g,84.38mmol)的甲醇(250mL)溶液中，加入三乙胺(43g,424.94mmol)和Pd(dppf)Cl2(3g,4.10mmol)，混合物于一氧化碳环境下120℃搅拌12h。反应完毕，过滤，加入2.5％碳酸氢钠水溶液(150mL x 3)洗涤，水相pH调节至2-3，乙酸乙酯(150mL x 3)萃取，无水硫酸钠干燥,浓缩得到粗品KH33-2(14g,收率76.8％)。LCMS(ESI,m/z):215.2[M-H]-.
第二步:向中间体KH33-2(5.5g,25.46mmol)的二氯甲烷(20mL)溶液中，加入中间体Int 2a(8.5g,25.22mmol)、2-氯-1-甲基吡啶碘化物(9.6g,37.64mmol)和DIEA(6.5g,50.29mmol)，混合物在室温下搅拌12h。反应完毕，用水(70mL)稀释，二氯甲烷(50mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩，粗品经硅胶柱色谱纯化得到KH33-3(9g,收率66.19％)。LCMS(ESI,m/z):434.8[M+H-100]+.
第三步:向中间体KH33-3(9g,16.75mmol)的乙酸乙酯(50mL)溶液中，加入盐酸乙酸乙酯(3M,50mL)，混合物室温搅拌12h，反应完毕，过滤，干燥滤饼，得到KH33-4(7g,收率96.0％)。LCMS(ESI,m/z):437.1[M+H]+.1H NMR(400MHz, DMSO)δ7.78(dd,J＝8.9,5.7Hz,1H),7.66(d,J＝8.2,4.7Hz,2H),7.26(dd,J＝8.9,6.3Hz,1H),4.85(s,2H),3.89(s,3H).
第四步：向中间体KH33-4(6.8g,15.56mmol)的DMSO(70mL)溶液中，加入DIEA(14mL,77.8mmol)，反应在150℃下搅拌2h，反应完毕用水(100mL)稀释，乙酸乙酯(50mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得到KH33-5(4.7g,收率72.7％)。LCMS(ESI,m/z):416.9[M+H]+.1H NMR(400MHz,DMSO)δ7.77(dd,J＝8.9,5.7Hz,1H),7.57(s,1H),7.29(dd,J＝8.6,6.4Hz,1H),7.12(d,J＝10.4Hz,1H),5.05(s,2H),3.87(s,3H).
第五步:向中间体KH33-5(800mg,2mmol)和中间体Int 3(592mg,2.9mmol)的DMF(12mL)溶液中，加入碳酸钾(550mg,4mmol)和碘化钠(60mg,0.4mmol)，50℃搅拌36h。反应完毕，加入水(8mL)和乙酸乙酯(8mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH33-6(300mg,收率32.1％)。LCMS(ESI,m/z):484.8[M+H]+.1H NMR(400MHz,CDCl3)δ7.68(s,1H),7.40(d,J＝9.6Hz,1H),7.33-7.27(m,1H),7.11-7.05(m,1H),5.18(s,2H),4.84-4.74(m,1H),4.63-4.54(m,1H),4.47-4.38(m,1H),4.19-4.14(m,1H),3.98-3.92(m,4H),2.67-2.57(m,1H),2.42-2.30(m,1H).
第六步:向中间体KH33-6(130mg,0.267mmol)和联硼酸频哪醇酯(101mg,0.4mmol)的二氧六环(2mL)溶液中，加入乙酸钾(52mg,0.534mmol)与Pd(dppf)Cl2(20mg,0.027mmol)，氮气氛围下于80℃搅拌2.5h，得到中间体KH33-7。LCMS(ESI,m/z):535.1[M+H]+.
第七步:向中间体KH33-7的反应溶液中，加入水(0.03mL)，中间体Int 1a(83mg，0.267mmol),碳酸钾(73.6mg,0.543mmol)与Pd(dppf)Cl2(19mg,0.0267mmol)，氮气氛围90℃搅拌2.5h。反应完毕，浓缩，经硅胶柱纯化得到KH33-8(70mg,收率38.7％)。LCMS(ESI,m/z):632.9[M+H]+.
第八步:向中间体KH33-8(70mg,0.15mmol)的乙腈(2mL)溶液中，加入的TBD水溶液(0.9M，0.3mL)，室温搅拌2h。反应完毕，加入水(5mL)，滴加乙酸水溶液使溶液调成酸性，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，粗品经制备高效液相色谱纯化得到KH33。LCMS(ESI,m/z):658.8[M+H]+.1H NMR(400MHz,DMSO)δ13.60(s,1H),7.96(s,1H),7.94-7.83(m,2H),7.77-7.69(m,3H),7.48(d,J＝6.1Hz,1H),7.27(d,J＝10.1Hz,1H),7.17(dd,J＝11.3,6.0Hz,1H),6.97(d,1H),5.57(s,2H),5.23(dd,J＝3.6Hz,2H),4.74-4.64(m,1H),4.49-4.30(m,3H),4.27-4.18(m,1H),2.60-2.56(m,1H),2.40-2.30(m,1H).
实施例34：(S)-2-(4-(6-((6-氯-4-甲氧基吡啶-3-基)甲氧基)-5-氟吡啶-2-基)-2,5-二氟苄基)-4-氟-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH34)的制备
第一步:向起始物料KH34-1(136mg,0.78mmol)的1,4-二氧六环(2mL)溶液中，加入中间体KH31-3(200mg,0.78mmol)和叔丁醇钾的四氢呋喃溶液(1.5mL,1.0M)，混合物45℃搅拌30min，反应完毕，浓缩，粗品经硅胶柱纯化得到KH34-2(160mg,收率59.0％)。LCMS(ESI,m/z):348.8[M+H]+.1H NMR(400MHz,CDCl3)δ8.37(s,1H),7.25-7.20(m,1H),7.04(dd,J＝8.1,2.7Hz,1H),6.86(s,1H),5.42(s,2H),3.93(s,3H).
第二步:向中间体KH33-7反应溶液中，加入KH34-2、水(0.02mL)，碳酸钾(73mg,0.543mmol)与Pd(dppf)Cl2(20mg,0.03mmol)，氮气氛围90℃搅拌2.5h。反应完毕，加入水(5mL)，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，经硅胶柱纯化得到KH34-3(110mg,收率61.1％)。。LCMS(ESI,m/z):672.9[M+H]+.
第三步:向中间体KH34-3(100mg,0.15mmol)的乙腈(2mL)溶液中，加入的TBD水溶液(0.9M，0.4mL)，室温搅拌2h。反应完毕，加入水(5mL)，滴加乙酸水溶液使溶液调成酸性，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，得粗品经制备高效液相色谱纯化得到KH34。LCMS(ESI,m/z):658.8[M+H]+.1H NMR(400MHz,DMSO)δ13.57(s,1H),8.32(s,1H),7.96(s,1H),7.84-7.75(m,2H),7.48(d,J＝9.7Hz,1H),7.29-7.24(m,2H),7.19(dd,J＝11.2,5.9Hz,1H),5.47(s,2H),5.23(dd,J＝3.5Hz,2H),4.73-4.65(m,1H),4.47-4.32(m,3H),4.25-4.18(m,1H),3.93(s,3H),2.60-2.55(m,1H),2.39-2.31(m,1H).
实施例35：(S)-2-(4-(6-((4-氰基-2-甲氧基苄基)氧基)-5-氟吡啶-2-基)-2,5-二氟苄基)-4-氟-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH35)的制备
第一步：向中间体Int 1e2(215mg,1.32mmol)的二氧六环(10mL)溶液中加入t-BuOK(185mg,1.65mmol)和原料KH35-1(230mg,1.10mmol)，于室温下反应2h。反应完全后,加入饱和氯化铵(10mL)淬灭反应,加入乙酸乙酯(10mL x 3)萃取有机相,饱和食盐水洗涤(20mL),无水硫酸钠干燥,真空减压浓缩后得粗品,粗品经正向硅胶纯化得到KH35-2(180mg,收率48.8％)。LCMS(ESI,m/z)：336.9[M+H]+。1H NMR(400MHz,CDCl3)δ7.59(d,J＝7.6Hz,1H),7.32-7.23(m,3H),7.13(s,1H),7.05(d,J＝8.0Hz,1H),5.50(s,2H),3.91(d,J＝2.0Hz,3H).
第二步：将中间体KH35-2(63mg,0.19mmol)溶于1,4-二氧六环(5mL)和水(0.5mL),将中间体KH33-7(120mg,0.19mmol)、碳酸钾(80mg,0.57mmol)和Pd(dppf)Cl2(15mg,0.02mmol)加入反应体系,在氮气气氛下升温至90℃,反应3h。反应完毕后,加入水(10mL)淬灭反应,加入乙酸乙酯(10mL x 3)萃取有机相,饱和食盐水(10mL)洗涤,无水硫酸钠干燥,真空减压浓缩后得粗品,粗品经正向硅胶纯化得到KH35-3(80mg,收率65.1％)。LCMS(ESI,m/z)：663.2[M+H]+。1H NMR(400MHz,CDCl3)δ7.69(s,1H),7.64(dd,J＝10.8,6.2Hz,1H),7.58(d,J＝7.8Hz,1H),7.47-7.44(m,1H),7.43-7.43(dd,J＝9.2,6.6Hz,2H),7.28(d,J＝8.0Hz,1H),7.15(s,1H),7.06(dd,J＝11.2,6.0Hz,1H),5.59(s,2H),5.25(s,2H),4.82(d,J＝4.8Hz,1H),4.59(dd,J＝14.2,7.8Hz,1H),4.48-4.41(m,1H),4.19(dd,J＝16.0,6.8Hz,1H),4.02-3.96(m,1H),3.94(s,6H),2.61(d,J＝6.0Hz,1H),2.44-2.32(m,1H).
第三步：向中间体KH35-3(80mg,0.12mmol)的乙腈(5mL)溶液中加入TBD(0.9M,1mL),室温下搅拌反应1h。反应完全后,加入饱和食盐水(10mL)淬灭反应,加入乙酸乙酯(10mL x 3)萃取有机相,饱和食盐水洗涤(10mL),无水硫酸钠干燥,真空减压浓缩后得粗品,粗品经反向C18纯化得到KH35。LCMS(ESI,m/z)：649.0[M+H]+。1H NMR(400MHz,DMSO)δ13.45(s,1H),7.97(s,1H),7.82(dd,J＝10.2,8.4Hz, 1H),7.64(dd,J＝10.6,6.2Hz,1H),7.56(d,J＝7.8Hz,2H),7.49-7.43(m,2H),7.27(d,J＝10.0Hz,1H),7.18(dd,J＝11.2,6.0Hz,1H),5.55(s,2H),5.31-5.16(m,2H),4.72-4.65(m,1H),4.49-4.30(m,3H),4.22(d,J＝14.0Hz,1H),3.89(s,3H),2.59-2.56(m,1H),2.40-2.33(m,1H).
实施例36：(S)-2-(4-(2-((4-氰基-2-甲氧苄基)氧基)嘧啶-4-基)-2,5-二氟苄基)-4-氟-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH36)的制备
第一步:向中间体KH33-7(150mg,0.28mmol)的1,4-二氧六环(4mL)和水(0.4mL)溶液中，加入中间体KH29-3b(106mg,0.33mmol)、碳酸钾(80mg,0.57mmol)和Pd(dppf)Cl2(32mg,0.04mmol)，混合物60℃搅拌2h，反应完毕，浓缩，粗品经硅胶柱纯化得到KH36-2(124mg,收率68.5％)。LCMS(ESI,m/z):646.1[M+H]+.
第二步：向中间体KH36-2(124mg,0.18mmol)的乙腈(2mL)溶液中，加入TBD(0.9M,0.6mL)，混合物在室温下搅拌2h。反应完毕用水(5mL)稀释，醋酸(1M)调节PH至5-6，乙酸乙酯(3mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经制备高效液相色谱纯化得到KH36。LCMS(ESI,m/z):632.2[M+H]+.1H NMR(400MHz,DMSO)δ8.74(d,J＝5.2Hz,1H),7.89(s,1H),7.84(dd,J＝10.4,6.1Hz,1H),7.60-7.52(m,3H),7.44(dd,J＝7.7,1.4Hz,1H),7.27(d,J＝10.3Hz,1H),7.21(dd,J＝11.2,5.9Hz,1H),5.50(s,2H),5.24(s,2H),4.73-4.63(m,1H),4.45-4.31(m,3H),4.25-4.14(m,1H),3.90(s,3H),2.58-2.54(m,1H),2.41-2.30(m,1H).
实施例37：(S)-2-(2，5-二氟-4-(6-((5-(三氟甲基)噻唑-2-基)甲氧基)吡啶-2-基)-苄基)-4-氟-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH37)的制备
第一步:在-78℃氮气气氛下，向原料KH37-1(700mg,3.02mmol)的乙醚(10mL)溶液中，滴加正丁基锂(1.7mL,2.72mmol)，-78℃下搅拌10min，然后向反应液中滴加DMF(0.47mL,6.03mmol)，反应液在氮气氛围下-78℃搅拌20min。反应完毕后，加入氯化铵水溶液(30mL)，乙酸乙酯(30mL x 3)萃取，饱和食盐水洗涤(30mL)，无水硫酸钠干燥，浓缩。得到黄色油状粗品KH37-2(500mg,91.50％)，该粗品直接用于下一步。LCMS(ESI,m/z):182.0[M+H]+.
第二步:向中间体KH37-2(500mg,2.76mmol)的乙醇(10mL)溶液中，加入NaBH4(208.86mg,5.52mmol)，室温下搅拌1.5h。反应完毕后，加入水(30mL)，乙酸乙酯(30mL x 3)萃取，饱和食盐水洗涤(30mL)，无水硫酸钠干燥，浓缩。得到黄色油状粗品KH37-3(400mg,79.12％)，该粗品直接用于下一步。LCMS(ESI,m/z):184.0[M+H]+.
第三步:向中间体KH37-3(400mg,2.18mmol)的乙腈(10mL)溶液中，加入化合物2-溴-6-氟吡啶(349.42mg,1.99mmol)和碳酸铯(905.67mg,2.78mmol)，95℃搅拌2h。反应完毕后，加入水(30mL)，乙酸乙酯(30mL x 3)萃取，饱和食盐水洗涤(30mL)，无水硫酸钠干燥，浓缩。经硅胶柱纯化得到KH37-4(260mg,收率38.61％)。LCMS(ESI,m/z):338.9[M+H]+.1H NMR(400MHz,CDCl3)δ8.07(s,1H),7.51(t,J＝7.8Hz,1H),7.17(d,J＝7.5Hz,1H),6.82(d,J＝8.2Hz,1H),5.68(s,2H).
第四步:向中间体KH33-7的反应液中加入中间体KH37-4(69.44mg,0.205mmol)，K3PO4(130.39mg,0.614mmol)，Pd(dppf)Cl2(13.22mg,0.0204mmol)和H2O(1mL)，反应混合液在N2氛围下于80℃搅拌2小时。反应完毕，加入水(15mL)稀释，乙酸乙酯(10mL x 3)萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤浓缩。粗品经硅胶柱纯化得到KH37-5(80mg,收率58.79％)。LCMS(ESI,m/z):665.1[M+H]+.
第五步：向中间体KH37-5(80mg,0.120mmol)的乙腈(2mL)溶液中，加入TBD(2mL,0.9M)，混合物在室温下搅拌30分钟。反应完毕，用柠檬酸将反应液pH调至5～6，加入水(15mL)稀释，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩。粗品经制备高效液相色谱纯化得到KH37。LCMS(ESI,m/z):649.1[M-H]-.1H NMR(400MHz,DMSO)δ13.57(s,1H),8.45(s,1H),7.97(s,1H),7.92(t,J＝7.9Hz,1H),7.80(dd,J＝10.8,6.2Hz,1H),7.54(d,J＝6.3Hz,1H),7.27(d,J＝10.1Hz,1H),7.19(dd,J＝11.3,5.9Hz,1H),7.04(d,J＝8.2Hz,1H),5.84(s,2H),5.28-5.18(m,2H),4.72-4.65(m,1H),4.46-4.33(m,3H),4.25-4.19(m,1H),2.59-2.54(m,1H),2.37-2.32(m,1H).
实施例38：(S)-2-(2，5-二氟-4-(6-((5-氯噻唑-2-基)甲氧基)吡啶-2-基)-苄基)-4-氟-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH38)的制备
第一步:在0℃下，向原料KH38-1(1g,6.78mmol)的四氢呋喃(10mL)溶液中缓慢加入NaBH4(384.53mg,10.16mmol)，室温搅拌1h。反应完毕后，加入水(30mL)，乙酸乙酯(30mL x 3)萃取，饱和食盐水洗涤(30mL)，无水硫酸钠干燥，浓缩。经硅胶柱纯化得到KH38-2(450mg,收率44.39％)。LCMS(ESI,m/z):150.2[M+H]+.1H NMR(400MHz,DMSO-d6)δ7.72(s,1H),6.22(t,J＝5.9Hz,1H),4.67(d,J＝5.9Hz,2H).
第二步:向中间体KH38-2(450mg,3.01mmol)的二氧六环(6mL)溶液中，加入化合物2-溴-6-氟吡啶(529.41mg,3.01mmol)，室温下向反应液中滴加叔丁醇钾(405.07mg,3.61mmol)的四氢呋喃(2mL)溶液，40℃搅拌1h。反应完毕后，加入1M的碳酸钾水溶液(30mL)，乙酸乙酯(20mL x 3)萃取，饱和食盐水洗涤(30mL)，无水硫酸钠干燥，浓缩。经硅胶柱纯化得到KH38-3(800mg,收率87.03％)。LCMS(ESI, m/z):306.9[M+H]+.1H NMR(400MHz,DMSO-d6)δ7.82-7.71(m,1H),7.41-7.36(m,2H),7.33-7.27(m,1H),4.96-4.90(m,1H),4.87(s,2H),4.77-4.68(m,1H),3.89(s,3H),3.88(s,3H).
第三步:向中间体KH33-7的反应液中加入中间体KH38-3(188.28mg,0.616mmol)，磷酸钾(196.19mg,0.924mmol)，Pd(dppf)Cl2(19.89mg,0.0308mmol)和水(1mL)，氮气氛围下80℃搅拌1h。反应完毕，加入水(20mL)，乙酸乙酯(10mL x3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩。粗品经硅胶板纯化得到KH38-4(170mg,收率87.45％)。LCMS(ESI,m/z):631.1[M+H]+.
第四步：向中间体KH38-4(170mg,0.269mmol)的乙腈(3mL)溶液中，加入TBD(3mL,0.9M)，混合物在室温下搅拌3小时。LCMS监测到少量产物生成，大量原料剩余，补加TBD(17mL,0.9M)，室温下搅拌过夜。反应完毕，用柠檬酸将反应液pH调至5～6，加入水(10mL)，乙酸乙酯(8mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩。粗品经制备高效液相色谱纯化得到KH38。LCMS(ESI,m/z):615.1[M-H]-.1H NMR(400MHz,DMSO)δ13.56(s,1H),7.97(s,1H),7.93-7.80(m,3H),7.53(d,J＝7.3Hz,1H),7.27(d,J＝10.1Hz,1H),7.19(dd,J＝11.1,5.9Hz,1H),6.99(d,J＝8.2Hz,1H),5.69(s,2H),5.30-5.17(m,2H),4.72-4.63(m,1H),4.47-4.31(m,3H),4.26-4.17(m,1H),2.59-2.53(m,1H),2.39-2.31(m,1H).
实施例39：(S)-2-(4-(6-((4-氰基-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-4-甲氧基-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH39)的制备
第一步:向原料KH39-1(900mg,3.61mmol)的DMF(10mL)溶液中，加入碘甲烷(614mg,4.32mmol)和碳酸铯(2.3g,7.05mmol)，室温搅拌1h。反应完毕，加入水(30mL)稀释，乙酸乙酯(20mL x 3)萃取，饱和食盐水(15mLx3)洗涤，无水硫酸钠干燥，过滤，浓缩，粗品经正相柱得到KH39-2(750mg,收率78.9％)。LCMS(ESI,m/z):No.[M+H]+.1H NMR(400MHz,DMSO)δ7.29(d,J＝8.9,1.3Hz,1H),7.25(s,1H),3.86(s,3H),3.83(s,3H).
第二步:向中间体KH39-2(750mg,2.85mmol)的甲醇(5mL)和水(1mL)溶液中，加入LiOH(340mg,14.19mmol)，室温搅拌4h。反应完毕，加入水(5mL)稀释，稀盐酸(1.0M)调节PH至3-4，乙酸乙酯(10mL x 3)萃取，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，浓缩，粗品得到KH39-3(700mg,收率99％)。LCMS(ESI,m/z):247.0[M-H]-.
第三步:向中间体KH39-3(700mg,2.82mmol)的甲醇(10mL)溶液中，加入TEA(570mg,5.63mmol)、和Pd(dppf)Cl2(204mg,0.27mmol)，反应在一氧化碳环境下120℃搅拌过夜。反应完毕，浓缩，加入乙酸乙酯(50mL)，0.25％碳酸氢钠水溶液(50mLx3)洗涤，水相用2.0M盐酸调节PH至2-3，加入乙酸乙酯(50mLx3)萃取，饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，浓缩得粗品KH39-4(600mg，93.3％)。LCMS(ESI,m/z):227.0[M-H]-.
第四步:向中间体KH39-4(569mg,2.49mmol)的DCM(7mL)溶液中，加入中间体Int 2a(840mg，2.49mmol)、2-氯-1-甲基吡啶碘化物(701mg,2.74mmol)和三乙胺(478mg，4.72mmol)，反应室温搅拌3h。反应完毕，浓缩，粗品经硅胶柱纯化得到KH39-5(500mg,收率36.7％)。LCMS(ESI,m/z):612.0[M+64]+.
第五步：向中间体KH39-5(500mg,0.91mmol)的乙酸乙酯(8mL)溶液中，加入盐酸乙酸乙酯(4M,8mL)，室温搅拌过夜。反应完毕，浓缩，粗品得到KH39-6(400mg,收率98.0％)。LCMS(ESI,m/z):448.8[M+H]+.
第六步:向中间体KH39-6(350mg,0.78mmol)的DMSO(15mL)溶液中，加入DIEA(306mg,2.36mmol)，反应150℃搅拌2h。反应完毕，加入水(30mL)稀释，乙酸乙酯(25mL x 3)萃取，饱和食盐水(30mL)洗涤，无水硫酸钠干燥，过滤，浓缩，粗品经正相柱得到KH39-7(90mg,收率27.0％)。LCMS(ESI,m/z):426.8[M+H]+.
第七步:向中间体KH39-7(60mg,0.14mmol)的DMF(1.5mL)溶液中，加入中间体Int 3(34mg,0.17mmol)和碳酸钾(39mg,0.28mmol)，反应50℃搅拌过夜。反应完毕，加入水(30mL)稀释，乙酸乙酯(20mL x 3)萃取，饱和食盐水(30mL)洗涤，无水硫酸钠干燥，过滤，浓缩，粗品经正相柱得到KH39-8(30mg,收率43.1％)。LCMS(ESI,m/z):499.1[M+H]+.
第八步:向中间体KH39-8(30mg,0.06mmol)的1,4-二氧六环(2mL)溶液中，加入联硼酸频哪醇酯(23mg,0.09mmol)、醋酸钾(12mg,0.12mmol)和Pd(dppf)Cl2(5mg,0.006mmol)，反应在氮气环境下80℃搅拌3h。反应完毕，体系直接用于下一步(32.8mg,100％)。LCMS(ESI,m/z):463.1[M-82+H]+.
第九步:向KH39-9(32.8mg,0.06mmol)的1,4-二氧六环(2mL)和水(0.2mL)溶液中，加入中间体Int 1a(19mg,0.06mmol)、碳酸钾(17mg,0.12mmol)和Pd(dppf)Cl2(5mg,0.006mmol)，反应在氮气环境下90℃搅拌过夜。反应完毕，浓缩，粗品经正相柱得到KH39-10(20mg,51.7％)。LCMS(ESI,m/z):644.9[M+H]+.
第十步：向中间体KH39-10(20mg,0.03mmol)的乙腈(1mL)溶液中，加入TBD(0.9M,0.5mL)，混合物在室温下搅拌1.5h。反应体系加水(5mL)稀释，乙酸(1M)调节PH至3-4，乙酸乙酯(5mL x 3)萃取，饱和食盐水(10mL)洗涤，无水硫酸钠干燥，过滤，浓缩，粗品经制备高效液相色谱纯化得到KH39。LCMS(ESI,m/z):631.1[M+H]+.1H NMR(400MHz,DMSO-d6)δ7.93-7.81(m,2H),7.76-7.68(m,3H),7.65(s,1H),7.47(d,J＝6.6Hz,1H),7.11-7.03(m,2H),6.97(d,J＝8.3Hz,1H),5.56(s,2H),5.16(s,2H),4.71-4.60(m,1H),4.43-4.28(m,3H),4.20-4.10(m,1H),3.91(s,3H),2.59-2.56(m,1H),2.38-2.32(m,1H).
实施例40：(S)-2-(4-(6-((4-氯-2-氟苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-4-甲氧基-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH40)的制备
采用实施例39类似的方法，以中间体KH39-9和Int 1f-1为起始物料，得到化合物KH40。LCMS(ESI,m/z):640.1[M+H]+.1H NMR(400MHz,DMSO-d6)δ13.34(s,1H),7.87-7.75(m,2H),7.64(s,1H),7.58(t,J＝8.2Hz,1H),7.47(t,J＝9.3Hz,2H),7.31(d,J＝8.2Hz,1H),7.10-7.02(m,2H),6.92(d,J＝8.3Hz,1H),5.46(s,2H),5.17(s,2H),4.71-4.60(m,1H),4.45-4.25(m,3H),4.20-4.10(m,1H),3.91(s,3H),2.59-2.52(m,1H),2.39-2.30(m,1H).
实施例41：(S)-2-(2，5-二氟-4-(6-((5-溴噻唑-2-基)甲氧基)吡啶-2-基)-苄基)-4-甲氧基-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH41)的制备
第一步:向原料KH41-1(500mg,2.60mmol)的四氢呋喃(5mL)和甲醇(0.3mL)溶液中，缓慢加入NaBH4(148mg,3.91mmol)，反应混合液在室温下搅拌2小时。反应完毕，加入水稀释，二氯甲烷(20mL x 3)萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥,浓缩。粗品经硅胶柱纯化得到KH41-2(240mg,收率47.50％)。LCMS(ESI,m/z):194.0[M+H]+.
第二步:向中间体KH41-2(240mg,1.24mmol)的二氯甲烷(5mL)溶液中，依次加入TEA(0.52mL,3.71mmol)和MsCl(0.28mL,3.71mmol)，反应液在室温下搅拌过夜。点板监测到有原料剩余，补加TEA(0.52mL,3.71mmol)和MsCl(0.28mL,3.71mmol)，室温下继续搅拌6小时。反应完毕，加入水(20mL)，二氯甲烷(15mL x  3)萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥,浓缩，得到KH41-3(336mg,收率100％)。
第三步:向(S)-2-(4-(6-羟基吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸甲酯(30mg,0.059mmol)的N,N-二甲基甲酰胺(1mL)溶液中，加入中间体KH41-3(31.9mg,0.117mmol)和Ag2CO3(64.69mg,0.234mmol)，反应混合液在50℃搅拌过夜。反应完毕，加入水(15mL)，乙酸乙酯(10mL x 3)萃取，有机相用水(15mL x 3)和饱和食盐水洗涤，无水硫酸钠干燥,浓缩。粗品用TLC板纯化得到KH41-4(12mg,收率29.76％)。LCMS(ESI,m/z):689.1[M+H]+.
第四步：向中间体KH41-4(12mg,0.017mmol)的乙腈(2mL)溶液中，加入TBD(1mL,0.9M)，反应混合液在室温下搅拌过夜。反应完毕，用柠檬酸将反应液pH调至3～4，加入水(10mL)，乙酸乙酯(10mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩。粗品经制备高效液相色谱纯化得到KH41。LCMS(ESI,m/z):673.0[M-H]-.1H NMR(400MHz,DMSO)δ7.84-7.77(m,2H),7.72(s,1H),7.61(s,1H),7.54(d,J＝7.2Hz,1H),7.24(s,1H),7.00(dd,J＝11.3,5.9Hz,1H),6.90(d,J＝8.2Hz,1H),5.69(s,2H),5.32(s,2H),4.82-4.80(m,1H),4.57-4.42(m,2H),4.37-4.30(m,1H),4.18-4.11(m,1H),4.00(s,3H),2.66-2.39(m,2H).
实施例42：(S)-2-(2，5-二氟-4-(6-((5-氯噻唑-2-基)甲氧基)吡啶-2-基)-苄基)-4-甲氧基-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH42)的制备
采用实施例39第九步、第十步类似的方法，以中间体KH39-9和KH38-3为起始物料，得到目标分子KH42。LCMS(ESI,m/z):629.1[M+H]+.1H NMR(400MHz,DMSO-d6)δ13.29(s,1H),7.92-7.82(m,3H),7.64(s,1H),7.53-7.50(m,1H),7.09-7.03(m,2H),6.98(d,J＝8.2Hz,1H),5.69(s,2H),5.16(s,2H),4.69-4.60(m,1H),4.44-4.26(m,3H),4.20-4.09(m,1H),3.91(s,3H),2.57-2.52(m,1H),2.39-2.27(m,1H).
实施例43：(S)-2-(2，5-二氟-4-(6-((5-(三氟甲基)噻唑-2-基)甲氧基)吡啶-2-基)-苄基)-4-甲氧基-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH43)的制备
采用实施例39第九步、第十步类似的方法，以中间体KH39-9和KH37-4为起始物料，得到目标分子KH43。LCMS(ESI,m/z):663.0[M+H]+.1H NMR(400MHz,MeOD-d4)δ8.21(d,J＝1.0Hz,1H),7.86-7.73(m,2H),7.65(s,1H),7.55(d,J＝6.5Hz,1H),7.23(s,1H),7.04-6.98(m,1H),6.95(d,J＝8.2Hz,1H),5.80(s,2H),5.32(s,2H),4.82-4.79(m,1H),4.59-4.51(m,1H),4.49-4.41(m,1H),4.39-4.30(m,1H),4.20-4.11(m,1H),4.01(s,3H),2.68-2.58(m,1H),2.49-2.37(m,1H).
实施例44：(S)-2-(4-(2-((4-氯-2-甲氧苄基)氧基)嘧啶-4-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH44)的制备
第一步：向原料KH44-1(1g,5.86mmol)的四氢呋喃(20mL)溶液中缓慢加入NaBH4(168.7mg,4.46mmol),于室温下反应2h。反应完全后,加入饱和氯化铵(30mL)淬灭反应,加入乙酸乙酯(30mL x 3)萃取有机相,饱和食盐水(50mL)洗涤,无水硫酸钠干燥,真空减压浓缩后得到KH44-2(1.03g)。LCMS(ESI,m/z)：155.2[M+H-18]+。1H NMR(400MHz,CDCl3)δ7.21(d,J＝8.0Hz,1H),6.93(dd,J＝8.0,1.8Hz,1H),6.86(d,J＝1.8Hz,1H),4.64(s,2H),3.85(s,3H).
第二步：向中间体KH44-2(500mg,2.90mmol)的四氢呋喃(10mL)溶液中加入t-BuOK(356mg,3.17mmol),室温搅拌半小时后加入化合物4-溴-2-(甲磺酰)嘧啶(626mg,2.64mmol)于0℃下反应2h。反应完全后,加入饱和氯化铵(30mL)淬灭反应,加入乙酸乙酯(20mL x 3)萃取有机相,饱和食盐水洗涤(40mL),无水硫 酸钠干燥,真空减压浓缩后得粗品,粗品经正向硅胶纯化得到KH44-3(400mg,收率41.8％)。LCMS(ESI,m/z)：330.8[M+H]+。1H NMR(400MHz,CDCl3)δ8.28(d,J＝4.8Hz,1H),7.40(d,J＝8.4Hz,1H),7.15(d,J＝4.8Hz,1H),6.94(dd,J＝8.2,1.8Hz,1H),6.88(d,J＝1.6Hz,1H),5.42(s,2H),3.84(s,3H).
第三步：将KH44-3(93mg,0.28mmol)溶于1,4-二氧六环(5mL)和水(0.5mL),将中间体KH2-8b(150mg,0.28mmol)，碳酸钾(78mg,0.56mmol)和Pd(dppf)Cl2(22mg,0.03mmol)加入反应体系,在氮气气氛下升温至90℃,反应3h。反应完毕后,加入水(10mL)淬灭反应,加入乙酸乙酯(10mL x 3)萃取有机相,饱和食盐水(10mL)洗涤,无水硫酸钠干燥,真空减压浓缩后得粗品,粗品经正向硅胶纯化得到KH44-4(110mg,收率61.1％)。LCMS(ESI,m/z)：637.2[M+H]+。1H NMR(400MHz,CDCl3)δ8.57(d,J＝5.2Hz,1H),8.00-7.94(m,2H),7.92(s,1H),7.84(d,J＝8.0Hz,1H),7.46(d,J＝3.8Hz,1H),7.41(d,J＝8.0Hz,1H),7.03-6.98(m,1H),6.93(d,J＝8.2Hz,1H),6.90(s,1H),5.49(s,2H),5.30(d,J＝3.6Hz,2H),4.79(d,J＝4.4Hz,1H),4.61-4.54(m,1H),4.45-4.39(m,1H),4.21-4.15(m,1H),3.98(d,J＝2.8Hz,1H),3.95(s,3H),3.87(s,3H),2.58(d,J＝5.6Hz,1H),2.44-2.33(m,1H).
第四步：向中间体KH44-4(110mg,0.17mmol)的乙腈(5mL)溶液中加入TBD(0.9M,1mL),室温下搅拌反应1h。反应完全后,加入饱和食盐水(10mL)淬灭反应,加入乙酸乙酯(10mL x 3)萃取有机相,饱和食盐水洗涤(10mL),无水硫酸钠干燥,真空减压浓缩后得粗品,粗品经反向C18纯化得到KH44。LCMS(ESI,m/z)：622.9[M+H]+。1H NMR(400MHz,DMSO)δ8.72(d,J＝5.2Hz,1H),8.12(s,1H),7.86(dd,J＝10.4,6.1Hz,1H),7.80(d,J＝8.0Hz,1H),7.69(dd,J＝8.2,1.2Hz,1H),7.53(dd,J＝5.0,1.8Hz,1H),7.41(d,J＝8.0Hz,1H),7.20-7.13(m,2H),7.02(dd,J＝8.0,2.0Hz,1H),5.40(s,2H),5.26(s,2H),4.68-4.60(m,1H),4.36-4.31(m,3H),4.23-4.16(m,1H),3.84(s,3H),2.58-2.51(m,1H),2.38-2.32(m,1H).
实施例45：(S)-2-(2，5-二氟-4-(6-((5-(三氟甲基)噻唑-2-基)甲氧基)吡啶-2-基)-苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH45)的制备
第一步:向原料KH45-1(1g,4.31mmol)的乙醚(15mL)溶液中，-78℃下加入正丁基锂(2.56mL,4.09mmol)，-78℃度搅拌10min。甲酸乙酯(638.56mg,8.62mmol)加入反应体系，室温搅拌2h。氯化铵溶液(30mL)淬灭反应，乙酸乙酯(30mL x 3)萃取，饱和食盐水(20mL)洗涤，无水硫酸钠干燥,浓缩，得到KH45-2(700mg,粗品)。LCMS(ESI,m/z):181.9[M+H]+.
第二步:向中间体KH45-2(1g,5.52mmol)的乙醇(10mL)溶液中，加入硼氢化钠(418mg,11.04mmol)，混合物室温搅拌2h。反应完毕，加入氯化铵(20mL)淬灭，乙酸乙酯(30mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥,浓缩，粗品得到KH45-3(0.8g,收率79.1％)。LCMS(ESI,m/z):183.9[M+H]+.
第三步:向KH45-3(200mg,1.08mmol)的二氯甲烷(3mL)溶液中，加入三乙胺(164mg,1.62mmol)和甲基磺酰氯(149mg,1.30mmol)，混合物室温搅拌2h。反应完毕，浓缩，粗品得到KH45-4(250mg,收率88.6％)。LCMS(ESI,m/z):261.8[M+H]+.
第四步：将中间体KH2-8b(164.48mg,0.32mmol)溶于1,4-二氧六环(1mL)和水(1mL)，依次将6-氯-2-羟基吡啶(41.6mg,0.32mmol)、K2CO3(122.88mg,0.96mmol)和Pd(dppf)Cl2(21.77mg,0.03mmol)加入反应体系，反应在90℃下搅拌3h。反应完毕，浓缩，粗品经硅胶柱纯化得到KH45-5(100mg,收率64.97％)。LCMS(ESI,m/z):482.0[M+H]+.
第五步:将中间体KH45-5(92mg,0.19mmol)溶于DMF(3mL)，依次将中间体KH45-4(100mg,0.38mmol)和碳酸银(211mg,0.76mmol)加入反应体系，碘化钠催化，50℃搅拌12h。反应完毕，加水(10mL)稀释，乙酸乙酯(5mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，粗品经硅胶柱纯化得到KH45-6(100mg,收率40.6％)。LCMS(ESI,m/z):646.9[M+H]+.
第六步：向中间体KH45-6(100mg,0.15mmol)的乙腈(2mL)溶液中，加入TBD(0.9M,0.5mL)，混合物在室温下搅拌2h。浓缩，得粗品经制备高效液相色谱纯化得到KH45。LCMS(ESI,m/z):633.1[M+H]+.1H NMR(400MHz,MeOD)δ8.19(s,1H),8.11(s,1H),7.89-7.86(m,1H),7.85-7.72(m,3H),7.51(d,J＝7.2Hz,1H),7.01(dd,J＝11.3,5.9Hz,1H),6.92(d,J＝8.2Hz,1H),5.77(s,2H),5.36(s,2H),4.74-4.80(m,1H),4.57-4.50(m,1H),4.46-4.35(m,2H),4.18(dd,J＝16.2,2.0Hz,1H),2.68-2.57(m,1H),2.51-2.37(m,1H).
实施例46：(S)-2-(2，5-二氟-4-(6-((5-(三氟甲基)噻唑-2-基)甲氧基)吡啶-2-基)-苄基)-4-氯-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH46)的制备
第一步:向原料KH46-1(5g,19.73mmol)的甲醇(70mL)溶液中，加入TEA(8.23mL,59.18mmol)和Pd(dppf)Cl2(1.26g,1.97mmol)，反应混合液在一氧化碳(气球压力)氛围下80℃搅拌过夜。反应完毕，将反应液浓缩，加入水(80mL)稀释， 乙酸乙酯(40mL x 3)萃取，用柠檬酸将水相pH调至1～2，乙酸乙酯(40mL x 3)萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥,浓缩，得到KH46-2(4.2g,收率91.53％)。LCMS(ESI,m/z):231.0[M-H]-.
第二步:0℃下，向中间体KH46-2(5.6g,24.08mmol)的二氯甲烷(50mL)溶液中，加入草酰氯(4.11mL,49.18mmol)和DMF(0.18mL,2.41mmol)，反应混合液在室温下搅拌30min。反应完毕，将反应液浓缩得到KH46-3(6g,收率100％)。
第三步:向中间体Int 2a(7.31g,21.67mmol)和三乙胺(10.04mL,72.23mmol)的二氯甲烷(50mL)溶液中，加入中间体KH46-3(6g,23.90mmol)的二氯甲烷溶液，反应混合液在室温下搅拌1h。反应完毕，加入水(80mL)稀释，二氯甲烷(30mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩，得到KH46-5(11g,收率82.81％)。LCMS(ESI,m/z):452.9[M-100+H]+.
第四步:向中间体KH46-4(11g,19.94mmol)的乙酸乙酯(20mL)溶液中，加入盐酸-乙酸乙酯(50mL,4M)溶液，反应混合液在室温下搅拌6h。反应完毕，浓缩。粗品经过滤纯化得到KH46-5(4.9g,收率54.42％)。LCMS(ESI,m/z):452.9[M+H]+.
第五步：中间体KH46-5(4.9g,10.85mmol)的NMP(50mL)溶液150℃下搅拌4.5h。反应完毕，加入水(100mL)稀释，有固体析出，过滤，将滤饼干燥，得到KH46-6(4.4g,收率93.96％)。LCMS(ESI,m/z):433.0[M+H]+.
第六步:向中间体KH46-6(340mg,0.788mmol)和Int 3(311.95mg,1.58mmol)的DMF(6mL)溶液中，加入K2CO3(326.61mg,2.36mmol)，反应混合液在50℃搅拌过夜。反应完毕，加入水(20mL)稀释，乙酸乙酯(15mL x 2)萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤浓缩。粗品经硅胶柱纯化得到KH46-7(130mg,收率32.89％)。LCMS(ESI,m/z):502.9[M+H]+.
第七步:向中间体KH46-7(130mg,0.259mmol)和联硼酸频哪醇酯(131.60mg,0.518mmol)的二氧六环(5mL)溶液中，加入Pd(dppf)Cl2(16.52mg,0.0259mmol)和KOAc(76.29mg,0.777mmol)，反应混合液在N2氛围下于90℃搅拌过夜。LCMS显示反应未完全。补加联硼酸频哪醇酯(100mg,0.393mmol)，Pd(dppf)Cl2(16mg,0.0251mmol)和KOAc(80mg,0.815mmol)，重新换气3次，继续反应5小时。反应完毕，直接用于下一步。LCMS(ESI,m/z):467.1,549.2[M+H]+.(主要生成硼酸)
第八步:向中间体KH46-8和中间体KH37-4(60mg,0.177mmol)的THF(4mL)和H2O(1mL)溶液中，加入K3PO4(92.83mg,0.437mmol)，Pd(dppf)Cl2(9.41mg,0.0146mmol)，反应混合液在N2氛围下于80℃搅拌1小时。反应完毕，加入水(15mL)稀释，乙酸乙酯(10mL x 3)萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤 浓缩。粗品经硅胶板纯化得到KH46-9(50mg,纯度34％,收率17.12％)。LCMS(ESI,m/z):681.0[M+H]+.
第九步：向中间体KH46-9(50mg,0.0734mmol)的乙腈(2mL)溶液中，加入TBD(1.5mL,0.9M)，混合物在室温下搅拌2小时。反应完毕，用柠檬酸将反应液pH调至5～6，加入水(10mL)稀释，乙酸乙酯(8mL x 3)萃取，饱和食盐水洗涤，无水硫酸钠干燥，浓缩。粗品经制备高效液相色谱纯化得到KH46。LCMS(ESI,m/z):667.0[M+H]+.1H NMR(400MHz,DMSO)δ7.31(s,1H),7.10(s,1H),6.96-6.86(m,2H),6.81(s,1H),6.66(d,J＝7.5Hz,1H),6.16(dd,J＝11.2,6.0Hz,1H),6.06(d,J＝8.2Hz,1H),4.90(s,2H),4.46(s,2H),3.92-3.88(m,1H),3.69-3.61(m,1H),3.58-3.47(m,2H),3.34-3.26(m,1H),1.83-1.69(m,1H),1.59-1.48(m,1H).
实施例47：2-(4-(6-((4-氰基-2-氯苄基)氧基)吡啶-2-基)-2,5-二氟苄基)-1-((氧杂环丁-2-基)甲基)-3-氧代-2,3-二氢-1H-吲唑-6-羧酸(KH47)的制备
采用实施例3相同的合成方法，以中间体Int 1f-5为原料与中间体KH2-8b反应，得到目标分子KH47。LCMS(ESI,m/z):617.5[M+H]+.1H NMR(400MHz,DMSO)δ13.02(s,1H),8.13(s,2H),7.91-7.79(m,3H),7.76-7.63(m,3H),7.48(d,J＝6.2Hz,1H),7.11(dd,J＝11.2,6.0Hz,1H),7.01(d,J＝8.2Hz,1H),5.58(s,2H),5.24(s,2H),4.64(d,J＝7.2Hz,1H),4.42-4.32(m,3H),4.20(d,J＝14.0Hz,1H),2.62-2.56(m,1H),2.41-2.27(m,1H).
实施例48：细胞实验
细胞实验一：Flp-in-293-GLP1R细胞激动活性测试
1、实验材料及仪器设备
表4
2、实验方法
2.1细胞培养及试剂配制
细胞株：Flp-in-293-GLP1R
完全培养基:DMEM+10％胎牛血清+1×青霉素链霉素+200μg/ml潮霉素
实验缓冲液:1×HBSS+20mM HEPES+0.1％BSA+500μM IBMX
2.2化合物对GLP1R受体激动活性的测定
a)将细胞消化，重悬到实验缓冲液中，种到384细胞培养板中，接种密度为20000每孔，接种体积为15μl每孔。
b)将化合物用实验缓冲液稀释，最大起始浓度1000pM，每次三倍稀释，共得到10个浓度梯度。
c)每孔加入5μl的化合物，37℃孵育30分钟。
d)冻融Eu-cAMP tracer和Ulight-anti-cAMP，用lysis buffer将其稀释。
e)加入10μl Eu-cAMP tracer至实验孔，然后加入10μl Ulight-anti-cAMP至实验孔中。
f)将反应板于室温200g离心30s，25℃静置1h后，利用Envision收集数据。
2.3数据分析：利用GraphPad非线性拟合公式计算化合物EC50。
3、实验结果
表5

细胞实验二：HEK293-hGLP1R/CRE-luc细胞激动活性测试
1、实验材料及仪器设备
表6
2、实验方法
2.1.细胞培养及试剂配置
1)细胞株：HEK293-Human GLP1R/CRE-luc
2)完全培养基:DMEM+10％胎牛血清+1×青霉素链霉素+400μg/mL G418
2.2.化合物对GLP1R激动活性测定
a)将细胞消化，重悬到完全培养基中，种到384细胞培养板中，接种密度为20000每孔，接种体积为25μl每孔，于37℃5％CO2孵育过夜。
b)将化合物用完全培养基稀释，最大起始浓度15000nM，每次四倍稀释，共得到10个浓度梯度。
c)每孔加入5μl的化合物，于37℃5％CO2孵育5小时。
d)向384孔检测板的每个孔中加入20μl荧光素酶检测试剂。
e)离心后直接在酶标仪Envision上读化学发光值。
2.3数据分析:利用GraphPad非线性拟合公式计算化合物EC50。
3.实验结果
表7
实施例49：化合物对PDE10A1的抑制作用
参照专利CN110325530A(说明书87页，实施例4A-01)制备出化合物PF06882961
1、实验材料及仪器设备
表8
2、实验步骤
2.1配制FAM cAMP工作液：60μl FAM-Cyclic-3′,5′-AMP储存液加入到5940μl的PDE分析缓冲液中。按照25μl/孔的体积加入到所有孔中。
2.2配制化合物溶液：先将待测化合物用DMSO溶成10mM的储液。化合物储液用DMSO，稀释成1000μM稀释液。再取5μl稀释液加入到45μl PDE分析缓冲液中，配制成100μM化合物工作液。按照取5μl/孔的体积加入到化合物孔中。对照孔中每孔加入5μl含10％DMSO的PDE分析缓冲液。
2.3配制PDE10A1溶液:用PDE分析缓冲液将PDE10A1重组酶储液稀释到15pg/μl，按照20μl/孔的取液量加入到所有化合物孔和Vehicle对照孔中。空白对照孔中加入20μl PDE分析缓冲液。
2.4室温孵育1h。
2.5配制结合试剂：取240μl结合试剂加入到23760μl结合试剂稀释剂中,混匀。按照100μl/孔取液量加入到所有孔中。
2.6室温反应1h。
2.7在多功能酶标仪上读取荧光值FP(激发波长485±5nm，发射波长528±5nm)。
2.8数据归一化到％抑制，IC50值计算使用Graphpad 4参数logistic方程计算。原始数据计算公式：
％抑制率＝(FPV-FPS)/(FPV-FPB)×100％
FPS＝样品荧光值；FPV＝Vehicle对照荧光值；FPB＝空白对照荧光值.
3、实验结果
表9
实施例50：hERG抑制测试
实验采用手动膜片钳的方法评估化合物对稳定表达在体外CHO细胞系的hERG(人ether-à-go-go相关基因)钾通道(IKr，快速激活延迟整流心脏钾电流)电流的浓度-反应关系。CHO hERG细胞来源于Sophion生物科学公司(巴勒鲁普，丹麦)。
本试验中，供试品将首先溶解至适当的溶剂中，然后按0.1％的比例用ECS稀释成不同浓度的供试品工作溶液：0.37、1.1、3.3、10和30μM。每个浓度重复测定两个细胞。全细胞膜片钳技术下记录hERG电流，记录温度为室温。膜片钳放大器输出信号通过数模转换以及2.9KHz低通滤波。数据记录用Patchmaster Pro软件采集。对于每个细胞，每一个浓度的供试品的抑制百分比由记录到的电流反应用以下公式算出(1-供试品/阳性对照灌流后记录到的尾峰值电流/溶媒对照灌流记录到的尾峰值电流(起始电流))×100％.对于每一个浓度记录到所有的细胞抑制百分比取均值，IC50值由Hill拟合的方法由浓度效应曲线中得出。
所测试化合物中，化合物KH03-2、KH13、KH15-2对hERG的抑制IC50均大于30μM。
实施例51：基因工程hGLP1R小鼠葡萄糖耐受实验
实验动物：基因工程hGLP1R小鼠采购自上海南方模式，周龄6-8周，雄性，体重25-30克。检疫期3天，常规健康检查由兽医完成，表现异常的动物在试验前剔除。检疫合格的动物在尾部标记单独的动物号，饲养在AAALAC认证的动物中心的SPF级恒温恒湿的层流清洁房间内，单笼饲养(每只笼具具有笼具标签，标明动物数量、性别、品系、接收时间、组别以及实验开始时间)。饲养室温度22-25℃，湿度40-70％，灯光12小时明暗交替(笼具：由聚碳酸酯制成，软制玉米芯高压消毒清洁垫料，每周更换两次。饲料和饮水：清洁级鼠料，购自北京科奥协力饲料有限公司。饮用水经过高压灭菌处理，食物经过钴60射线照射。动物可以自由摄取无菌食物和饮水)。
实验方法：动物适应性饲养7天后，于给药前一天，按体重将动物随机分组。在禁食过夜(17:00-09:00)后，空白给药组和其它给药组动物提前30分钟给药(空白给药组溶媒采用生理盐水；其它给药组溶媒均采用5％DMSO：20％Solutol：75％生理盐水，配置成澄清溶液)，口服给药，0h测量血糖基值后口服葡萄糖(2g/kg)，在15、30、60、120分钟取小鼠尾尖部静脉血，通过Accu-Chek Guide型血糖仪获得小鼠血糖读数，绘制血糖变化曲线，计算AUC面积。
数据统计：本实验所有数据的统计分析均使用GraphPad Prism 8.0.软件进行。所有数值以均数±标准差(Mean±SD)表示。采用单因素方差分析法(One-way ANOVA)比较组间差异。对于所有分析，P<0.05认为有统计学意义。测试结果如附图1和2所示(**表示与空白溶剂对照组相比，P<0.01)。
实施例52：高脂饮食和STZ联合诱导的hGLP1R转基因小鼠II型糖尿病模型 (HFD-STZ)实验
实验方法：10-12周龄的雄性hGLP1R转基因小鼠(购自上海南方模式生物科技股份有限公司)通过高脂饲料(货号D12492，Research Diets公司)诱导7周，第5、6周每周一次腹腔注射链脲佐菌素(STZ)诱导II型糖尿病模型；10-12周龄的正常雄性C57BL/6小鼠为正常对照组(空白组)，给予普通饲料。之后溶媒适应2周，模型对照组小鼠根据体重和空腹血糖水平分成3组，分别给予溶媒(BID，经口灌胃)、PF06882961(阳性药，15mg/kg，BID，经口灌胃)和受试物KH15-2(15mg/kg，BID，经口灌胃)；正常对照组给予溶媒(BID，经口灌胃)；每组8只，除正常对照组给予普通饲料，其余组给予HFD直至实验结束。治疗期间，监测动物的体重以及随机血糖，试验终点对所有动物做口服糖耐量实验。
实验结果：实验期间/终点动物重、随机血糖、空腹血糖、糖耐受结果分别如附图3、4和5所示。对于所有分析，P<0.05认为有统计学意义。(#代表P<0.05，##代表P<0.01，###代表P<0.001)。
结论：KH15-2对HFD+HTZ诱导的hGLP1R小鼠Ⅱ型糖尿病具有良好的治疗作用，能够降低模型小鼠的体重、随机血糖水平和空腹血糖水平，增强葡萄糖耐受。
实施例53:模拟空腹肠液溶解度实验
参照专利CN114761395A(说明书61页，实施例86)制备出对照化合物
实验过程：
1)储备液的配制：
用DMSO配制待测物和对照药双氯芬酸钠的10mM储备液。
2)表观溶解度的测定
取30μL 10mM待测物和对照药储备液，以指定顺序加到对应96孔板的对应位置。在样品板的对应小瓶加入970μL的FaSSIF溶液。实验为双平行。在每个小瓶中加一根搅拌棒，并盖上聚四氟乙烯/硅有机树脂瓶塞。随后将样品盘放进Eppendorf Thermomixer Comfort，以1100转的转速在25摄氏度条件下震荡2个小时。2小时后，去除瓶塞，用磁铁吸走搅拌棒，然后从样品板转移样品至过滤板。用真空泵产生负压，过滤样品。转移5μL滤液到新的样品板，然后加入5μL DMSO和490μL乙腈-水(1:1,v/v)。将 200μL稀释剂转移到上样盘中进行LC-MS/MS分析。稀释倍数可能因待测物溶解性的大小或其液质响应信号强弱而调整。
3)标准品溶液的配制
转移10mM DMSO储备液15μL至一空板中，加入485μL DMSO配成300μM的标准品溶液。转移5μL 300μM的标准品溶液到另一空板中，再加入5μL FaSSIF溶液和490μL乙腈-水(1:1,v/v)，配制成终浓度为3μM的标准品溶液，用于进样分析。标准样品的浓度可能会根据LC-MS信号响应而调整。
4)样品分析
采用LC-MS/MS法进行样品分析。
5)数据分析
采用Microsoft Excel进行数据计算。采用LC-MS/MS法分析滤液样品，并用单点定量法对样品中待测药物进行定性与定量分析。对照药和待测药的溶解度值计算公式如下：
DF：Dilution Factor，稀释倍数。
实验结果、结论：不同化合物测试溶解度结果如表10所示，化合物KH15-2、KH33、KH39、KH44的溶解度均显著优于对照化合物。
表10
实施例54：SD大鼠口服药代动力学实验
实验方法：实验前，动物禁食过夜(>12h)，不禁水。通过灌胃给予SD大鼠本发明的化合物澄清溶液，药物溶媒为5％DMSO∶10％Solutol∶85％生理盐水，给药剂量为5mg/kg。给药后15min、0.5、1、2、4、6、8、24h分别经颈静脉采血，EDTA-K2抗凝后(血样采集后放置冰浴)于4℃，6000g，离心5min，分离血浆，血浆于-70℃保存待测。采用LC-MS/MS法测定血浆内指定化合物药物浓度；Winnolin 8.3非房室模型计算主要药动学参数。
实验结果/结论：PK主要参数见表11，从表中可以看出，化合物KH15-2的口服暴露量(Cmax和AUC)和生物利用度显著优于对照化合物。
表11

Claims (18)

  1. 如式I所示的化合物或其药学上可接受的盐：
    其中R1选自氢原子、卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、氰基、氨基、硝基、羟基、环烷基、杂环基、芳基和杂芳基，其中所述的烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、环烷基、杂环基、芳基和杂芳基任选地被选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、氰基、氨基、硝基、羟基、羟烷基、环烷基、杂环基、芳基和杂芳基中的一个或多个取代基所取代；
    R2选自氢原子、烷基、芳基或杂芳基，其中所述的烷基、芳基或杂芳基任选被选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、氰基、氨基、硝基、羟基、羟烷基、环烷基、杂环基、芳基和杂芳基中的一个或多个取代基所取代，所述环烷基和杂环基任选被卤素、烷基、氰基中的一个或多个取代基所取代；
    R3在每次出现时相同或不同，且各自独立地选自氢原子、卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、羟烷基、氰基、氨基、硝基、羟基、环烷基、杂环基、芳基和杂芳基；
    R5选自环烷基、杂环基、芳基和杂芳基，其中所述的环烷基、杂环基、芳基和杂芳基任选地被选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、氰基、氨基、硝基、羟基、羟烷基、环烷基、杂环基、芳基和杂芳基中的一个或多个取代基所取代；
    X选自N、O或S；
    L为直链或支链烷基；
    Z0、Z1、Z2、Z3、Z4、Z5和Z6可以相同或不相同，各自独立地选CR4或氮原子；
    m、n、p为0-4的整数。
  2. 如权利要求1所述的式I化合物或其药学上可接受的盐，其特征在于R1、R3各自独立地选自氢原子、卤素、C1-6烷基、C1-6烷氧基、卤代C1-6烷基、卤代C1-6烷氧基、氰基、氨基、硝基、羟基、羟C1-6烷基、C3-8环烷基和C3-20杂环基。
  3. 如权利要求1所述的式I化合物或其药学上可接受的盐，其特征在于R2为被烷基、环烷基、杂环基取代的C1-6烷基，所述杂环基进一步优选为杂原子为O，任选地被选自卤素、烷基、烷氧基、卤代烷基、卤代烷氧基、氰基、氨基、硝基、羟基中的一个或多个取代基所取代。
  4. 如权利要求1所述的式I化合物或其药学上可接受的盐，其特征在于R5选自芳基或杂芳基，所述芳基或杂芳基任选地被烷基、烷氧基、卤代烷基、氰基、卤素所取代。
  5. 如权利要求1所述的式I化合物或其药学上可接受的盐，其特征在于所述化合物具有如式 II所示的结构：
    Z0选自C或N；
    Z2选自CR4或N，R4选自H或CH3；
    Z4、Z5或Z6选自C或N；
    R2选自杂环基或C1-6烷基，所述杂环基优选为任选取代的C3-8含氧杂环基，所述C1-6烷基优选被CH2OCH3、取代的C1-6烷基；
    R3在每次出现时相同或不同，各自独立地选自氢、F或甲基，n选自1或2的整数；
    R5选自取代或未取代的芳基或杂芳基，所述芳基或杂芳基任选被卤素、CN、OC1-6烷基、-C(＝O)C1-6烷基、-C(＝O)-N(C1-6烷基)2、C1-6卤代烷基所取代；
    L为CH2，p为1-4的整数。
  6. 如权利要求1所述的式II化合物，其特征在于R5中所述芳基或杂芳基选自：
    其中X0、X1、X2、X3和X4各自独立的选自C、N、O和S；
    优选X0选自C或N；
    进一步优选X1、X2、X3和X4至少其中一个是N；
    上述芳基或杂芳基任选被H、卤素、氰基、氨基、硝基、羟基、C1-6烷基、C1-6烷氧基、C3-8环烷基、卤代C1-6烷基、卤代C1-6烷氧基、羟C1-6烷基所取代。
  7. 如权利要求5所述的式II化合物或其药学上可接受的盐，其特征在于，所述化合物具有式III所述的结构：

    Z0选自C或N；
    Z2选自CR4或N，R4选自H或CH3；
    Z4、Z5或Z6选自C或N；
    R3选自F，n选自1或2的整数；
    R5选自如下芳基或杂芳基：
    其中X0选自C或N，
    所述芳基或杂芳基可以任选被F、Cl、CN、OCH3、CF3取代；
    R6选自-CH2OCH3、
    R10选自H、卤素、CN、OCH3、或CH2CF2。
  8. 如权利要求7所述的式III化合物或其药学上可接受的盐，其特征在于所述化合物具有式IV所示的结构：
    Z0选自C或N；
    Z4、Z5或Z6选自N或C；
    R3是F，n＝2；
    R5选自如下基团：
    其中X0选自C或N，
    R7选自F或甲氧基；R8选自Cl或CN；
    R9选自F或H；R11选自Cl或CN；
    R12选自卤素或CF3；
    R6选自
  9. 如权利要求8所述的式IV化合物或其药学上可接受的盐，其特征在于所述化合物具有式V所述的结构：
    R3选自F，n＝2；
    Z6选自C或N；
    R5选自如下基团：
    其中X0选自C或N，
    R7选自F或甲氧基；R8选自Cl或CN；
    R9选自F或H；R10选自Cl或CN；
    R12选自卤素或CF3；
    R6选自优选
  10. 如权利要求1所述的式I化合物或其药学上可接受的盐，其特征在于所述化合物具有式VI所述的结构：
    其中，
    R13选自氢、卤素、C1-3烷基、C1-3卤代烷基或C1-3烷氧基；
    Z4、Z5或Z6选自N或C；
    X0选自C或N；
    R7、R8、R14、R15选自氢、卤素、氰基、C1-3烷基、C1-3卤代烷基或C1-3烷氧基。
  11. 如权利要求10所述的式VI化合物或其药学上可接受的盐，其特征在于所述化合物具有式VII所述的结构：
    其中，
    R13选自氢、卤素、C1-3烷基、C1-3卤代烷基或C1-3烷氧基；
    Z6选自N或C；
    R7、R8选自氢、卤素、氰基、C1-3烷基、C1-3卤代烷基或C1-3烷氧基。
  12. 如权利要求11所述的式VI化合物或其药学上可接受的盐，其特征在于：
    R7选自卤素或甲氧基；
    R8选自卤素或CN。
  13. 如权利要求12所述的式VI化合物或其药学上可接受的盐，其特征在于：
    R7选自F或甲氧基；
    R8选自Cl或CN。
  14. 如权利要求1所述的式I化合物或其药学上可接受的盐，其特征在于所述化合物选自下述具体化合物：



  15. 一种药物组合物，其特征在于包括权利要求1-14之一所述的化合物或其可药用的盐和药学上可接受的载体。
  16. 权利要求1-14之一所述的化合物或权利要求15所述的药物组合物在制备用于激动GLP-1受体的药物中的用途。
  17. 如权利要求16所述的用途，其特征在于，所述用途选自用于制备治疗和/或预防I型糖尿病、II型糖尿病、营养不良相关性糖尿病、糖尿病并发症、肥胖症、高血糖症、葡萄糖耐受不良、心血管疾病、高脂血症、脑梗塞、中风、非酒精性脂肪性肝炎(NASH)、帕金森病、痴呆、胰岛素抗性和肝脏胰岛素抗性的药物中的用途。
  18. 如权利要求17所述的用途，其特征在于，所述用途选自用于制备治疗和/或预防I型糖尿病、II型糖尿病、肥胖症、糖尿病并发症、非酒精性脂肪性肝炎和心血管疾病的药物中的用途。
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