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WO2025209499A1 - POLθ INHIBITOR AND PHARMACEUTICAL COMPOSITION THEREOF AND USE - Google Patents

POLθ INHIBITOR AND PHARMACEUTICAL COMPOSITION THEREOF AND USE

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Publication number
WO2025209499A1
WO2025209499A1 PCT/CN2025/086748 CN2025086748W WO2025209499A1 WO 2025209499 A1 WO2025209499 A1 WO 2025209499A1 CN 2025086748 W CN2025086748 W CN 2025086748W WO 2025209499 A1 WO2025209499 A1 WO 2025209499A1
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WIPO (PCT)
Prior art keywords
mmol
methoxy
reaction
tetrahydrobenzo
thiazol
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/CN2025/086748
Other languages
French (fr)
Chinese (zh)
Inventor
陆平波
杨佳乐
祝辉
王敏超
邓民杰
王知
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Alicorn Pharmaceutical Co Ltd
Original Assignee
Alicorn Pharmaceutical Co Ltd
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Publication of WO2025209499A1 publication Critical patent/WO2025209499A1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • 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
    • A61K31/438—The ring being spiro-condensed with carbocyclic or heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • 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
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • 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
    • A61K31/445—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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • 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
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • 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
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A61K31/502—Pyridazines; Hydrogenated pyridazines ortho- or peri-condensed with carbocyclic ring systems, e.g. cinnoline, phthalazine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
    • C07D277/60—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings condensed with carbocyclic rings or ring systems
    • C07D277/62—Benzothiazoles
    • C07D277/68—Benzothiazoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 2
    • C07D277/82—Nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—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
    • C07D417/02—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 two hetero rings
    • C07D417/04—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 two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—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
    • C07D417/02—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 two hetero rings
    • C07D417/12—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 two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • the present invention belongs to the field of medicinal chemistry, and specifically relates to a Pol ⁇ inhibitor or its stereoisomers, tautomers, and pharmaceutically acceptable salts, as well as to a pharmaceutical composition of the compound and its use in treating cancer.
  • DNA damage repair pathways include base excision repair (BER), nucleotide excision repair (NER), DNA mismatch repair (MMR), and DNA double-strand break (DSB) repair.
  • Double-strand DNA damage repair pathways mainly include homologous recombination (HR), non-homologous end joining (NHEJ), and microhomology-mediated end joining (alt-EJ or MMEJ).
  • Homologous recombination mainly relies on homologous chromosomes to guide the correction of damaged DNA, while non-homologous end joining directly connects the two broken DNA chains together to achieve the purpose of repair.
  • BRCA1 and BRCA2 the key proteins in homologous recombination, are two important tumor suppressor factors.
  • NHEJ non-homologous end joining
  • DNA polymerase ⁇ (Pol ⁇ or POLQ) is a synthetic lethal target of homologous recombination (HR) defects and plays an important role in the DNA damage response (DDR) pathway for double-strand breaks (DSBs).
  • HR DNA damage response
  • DSBs double-strand breaks
  • BRCA2 BRCA2 not only recruits the recombinase RAD51 to the DSB to promote HR but also inhibits repair pathways such as alt-NHEJ.
  • homologous recombination-mediated repair is impaired (HR deficiency), as in the case of BRCA1 or BRCA2 mutations, Pol ⁇ is highly expressed and directs DSB repair toward alt-NHEJ, initiating the DNA repair process of microhomology-mediated end joining (MMEJ).
  • MMEJ microhomology-mediated end joining
  • Pol ⁇ is expressed in only a few tissue types but is highly expressed in many cancer cells. Pol ⁇ can confer resistance to cancer therapies and promote the survival of abnormal cells lacking DNA damage repair pathways.
  • the primary function of DNA polymerase ⁇ (Pol ⁇ ) is to act as a reverse transcriptase. In healthy cells, Pol ⁇ acts as an RNA-mediated DNA repair enzyme. In cancer cells, Pol ⁇ is highly expressed and promotes cancer cell growth and drug resistance.
  • Documents such as CN114127062A, CN114667167A, WO2020160213, WO2022118210, WO2021123785, and WO2023134708 disclose DNA polymerase Theta (Pol ⁇ ) inhibitors.
  • the heterocyclic compounds provided by the present invention exhibit excellent Pol ⁇ inhibitory effects.
  • the present invention provides a compound represented by formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof:
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 are each independently selected from H, deuterium, halogen, amino, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy;
  • R 8 is selected from H, hydroxy, C 1 -C 6 alkoxy, NR a R b ;
  • R 9 is phenyl, heteroaryl, heterocyclyl, -O(CH 2 ) r -cycloalkyl, -O(CH 2 ) r -heterocycloalkyl, -O(CH 2 ) r -heteroaryl, and the phenyl, heteroaryl, heterocyclyl, -O(CH 2 ) r -cycloalkyl, -O(CH 2 ) r -heterocycloalkyl, -O(CH 2 ) r -heteroaryl may be optionally substituted with one or more R n ;
  • R n is selected from H, halogen, cyano, NR a R b , hydroxy, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, C 1 -C 6 hydroxyalkyl, C 1 -C 6 alkylamino, C 1 -C 6 haloalkoxy, -(CH 2 ) r -OC 1 -C 6 alkyl, -C(O)NR a R b , -S(O) 2 NR a R b , -NH(CH 2 ) r C 1 -C 6 haloalkyl, -NH(CH 2 ) r OH, -(CH 2 ) r OC 1 -C 6 haloalkyl, C 1 -C 6 haloalkyl;
  • R a and R b are each independently selected from absent, hydrogen, and C 1 -C 6 alkyl;
  • r is an integer from 0 to 3.
  • R 4 is selected from methoxy.
  • R 8 is selected from hydrogen, hydroxy, methoxy, amino, and aminomethyl.
  • the compounds of the present invention possess asymmetric centers. Compounds of the present invention containing asymmetrically substituted atoms can be separated into optically active or racemic forms. Those skilled in the art will appreciate how to prepare optically active forms, such as by racemate resolution or synthesis from optically active starting materials. Unless otherwise indicated with respect to specific stereochemistry or isomeric forms, the present invention encompasses all chiral, diastereoisomer, and racemic forms. Methods for preparing the compounds of the present invention and intermediates thereto are also intended to be included in the present invention. All tautomers of the compounds of the present invention are also intended to be included in the present invention.
  • the term "optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not occur.
  • “optionally substituted alkyl” refers to events or circumstances where the alkyl group may be substituted as well as instances where the alkyl group is not substituted.
  • substituted refers to a moiety having a substituent that replaces a hydrogen on one or more carbons of the main chain. It should be understood that “substituted” or “substituted by" includes implicit conditions, i.e., such substitution is consistent with the allowed valence of the substituted atom and the substituent, and the substitution produces a stable compound, for example, it will not spontaneously undergo transformations such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is intended to include all permissible substituents of an organic compound.
  • permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of an organic compound.
  • the permissible substituents can be one or more and the same or different.
  • a heteroatom such as nitrogen can have a hydrogen substituent and/or any permissible substituent of an organic compound as described herein that satisfies the valence of the heteroatom.
  • Substituents can include any substituent described herein, such as halogen, hydroxyl, alkyl, alkoxy, amino, cyano, heteroaryl, heterocyclic, etc. It will be understood by those skilled in the art that substituents may themselves be substituted, if appropriate.
  • halogen refers to fluorine, chlorine, bromine, or iodine, and particularly to fluorine, chlorine, and bromine, with fluorine and chlorine being particularly preferred.
  • cyano refers to a group in which a carbon atom is triple-bonded to a nitrogen atom.
  • alkoxy is a customary expression and refers to an alkyl group attached to the rest of the molecule via an oxygen atom, and refers to an alkyl group containing an oxygen atom. Examples of such groups include methoxy, ethoxy, or propoxy.
  • alkyl refers to an alkyl group having a specified number of carbon atoms, which is a linear or branched alkyl group, and which may include subgroups thereof.
  • C1 - C6 alkyl group it may also include sub-ranges of groups represented by C1 - C4 alkyl, C1 - C3 alkyl, C2 - C6 alkyl, C2 - C4 alkyl, etc., as well as specific groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, etc.
  • haloalkyl refers to an alkyl group as defined above that is attached to the rest of the molecule via a halo linkage.
  • a C 1-6 haloalkyl group refers to an alkyl group having 1 to 6 carbon atoms or 1 to 3 carbon atoms that is attached to the rest of the molecule via a halo linkage.
  • Preferred haloalkyl groups include, but are not limited to, -CH 2 Cl, -CHCl 2 , -CF 3 , and the like.
  • hydroxy or "hydroxyl,” by itself or in combination with other terms, means -OH.
  • Nitrogen and sulfur atoms may be optionally oxidized, and nitrogen atoms may be optionally quaternized.
  • the fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated or fully unsaturated.
  • Heterocyclic groups may be attached to any available nitrogen or carbon atom.
  • the heterocyclic radicals include heterocycloalkyl and the like.
  • heteroaryl refers to substituted and unsubstituted aromatic 5- or 6-membered monocyclic groups, 9- or 10-membered bicyclic groups, and 11- to 14-membered tricyclic groups having at least one heteroatom (O, S, or N) in at least one ring, the heteroatom-containing ring preferably having 1, 2, or 3 heteroatoms selected from O, S, and N.
  • Each ring of the heteroaryl containing heteroatoms can contain one or two oxygen or sulfur atoms and/or one to four nitrogen atoms, provided that the total number of heteroatoms in each ring is four or less, and each ring has at least one carbon atom.
  • the fused rings completing the bicyclic and tricyclic groups can contain only carbon atoms and can be saturated, partially saturated, or unsaturated.
  • the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen atom may optionally be quaternized.
  • a bicyclic or tricyclic heteroaryl must include at least one fully aromatic ring, but the other one or more fused rings may be aromatic or non-aromatic.
  • heteroaryl examples include, but are not limited to, furyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, 1H-tetrazolyl, oxadiazolyl, triazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzofuranyl, benzothienyl, benzotriazinyl, phthalazinyl, thianthracene, dibenzofuranyl, benzothiophenyl ...
  • Step 6 Synthesis of 2'-chloro-N-[6-(6-fluoro-4-methylpyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • 6-(6-Fluoro-4-methylpyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (90.0 mg, 0.34 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (95.1 mg, 0.34 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (2 mL), and N-methylimidazole (84.2 mg, 1.03 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (105.8 mg, 0.41 mmol, 1.2 eq) were added, followed by reaction at room temperature for 2.0 hours.
  • Step 5 Synthesis of 2'-chloro-N- ⁇ 6-[2-fluoro-4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl ⁇ -5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • N-methylimidazole (88.23 mg, 1.08 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (150.7 mg, 0.54 mmol, 1.5 eq) were then added and reacted at room temperature for 2 hours. After the reaction, preparative separation and purification were performed to obtain 2'-chloro-N- ⁇ 6-[2-fluoro-4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl ⁇ -5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (42.7 mg, 20.62% yield) as a white solid.
  • 1,4-Dioxyaspirin[4.5]dec-7-en-8-yl trifluoromethanesulfonate (8.19 g, 28.4 mmol, 1.0 eq) and (2,4,5-trifluorophenyl)boronic acid (5.0 g, 28.4 mmol, 1.0 eq) were added to 1,4-dioxane (80 mL), followed by cesium carbonate (27.78 g, 85.3 mmol, 3.0 eq) and water (20 mL). Finally, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (831 mg, 1.1 mmol, 0.04 eq) was added.
  • Step 6 Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-[6-(2,4,5-trifluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-[4,4'-bipyridine]-3-carboxamide
  • 1,4-Dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (2.0 g, 6.94 mmol, 1.0 eq) was dissolved in 1,4-dioxane (10 mL).
  • (4-Cyano-3-fluorophenyl)boronic acid (1.01 g, 6.94 mol, 1.0 eq)
  • cesium carbonate (6.79 g, 20.8 mmol, 3.0 eq)
  • deionized water 2 mL
  • [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (253.8 mg, 0.35 mmol, 0.05 eq) were added.
  • Step 5 Synthesis of 2'-chloro-N-(6-(4-cyano-3-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • Step 3 Synthesis of phenyl 4-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridyl]-3-carboxamido)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl]acetate
  • Step 4 Synthesis of 2'-chloro-N-[6-(4-hydroxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • Step 5 Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-[6-(4-methylthiophen-2-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-[4,4'-bipyridine]-3-carboxamide
  • Methyl 2'-(difluoromethyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylate (550 mg, 1.78 mmol, 1.0 eq) was dissolved in methanol (6 mL) and lithium hydroxide monohydrate (112.5 mg, 2.67 mmol, 1.5 eq) was added. The mixture was reacted at 60°C for 1 hour. After completion of the reaction, the reaction solution was cooled to room temperature, the pH was adjusted to neutral, and the mixture was extracted three times with ethyl acetate.
  • Step 3 Synthesis of 2'-(difluoromethyl)-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide
  • 6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine 25 mg, 0.108 mmol, 1.0 eq
  • N-methylimidazole (26.6 mg, 0.324 mmol, 3.0 eq)
  • N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (45.7 mg, 0.162 mmol, 1.5 eq) were added in sequence and reacted at room temperature for 2 hours.
  • 1,4-Dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (4.0 g, 13.87 mmol, 1.0 eq) and 4-fluorophenylboronic acid (2.64 g, 13.87 mmol, 1.0 eq) were dissolved in ultra-dry 1,4-dioxane (30.0 mL).
  • Deionized water (7.5 mL), cesium carbonate (13.56 g, 41.62 mmol, 3.0 eq), and dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (0.456 g, 0.624 mmol, 0.41 eq) were then added.
  • Step 5 Synthesis of 2'-chloro-N-[6-(4-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • Step 2 Synthesis of 5'-methoxy-2',6'-dimethyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide
  • 1,4-Dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (3.0 g, 10.4 mmol, 1.0 eq) and 4-(difluoromethyl)phenylboronic acid (2.64 g, 13.87 mmol, 1.0 eq) were dissolved in ultra-dry 1,4-dioxane (16.0 mL), and deionized water (4.0 mL), cesium carbonate (10.17 g, 31.2 mmol, 3.0 eq) and dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (228 mg, 0.312 mmol, 0.03 eq) were added.
  • Step 5 Synthesis of 2'-chloro-N- ⁇ 6-[4-(difluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl ⁇ -5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • Step 1 Synthesis of tert-butyl [4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)phenyl]carbamate
  • 1,4-Dioxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate (4 g, 13.9 mmol, 1.0 eq), ⁇ 4-[(tert-butoxycarbonyl)amino]phenyl ⁇ boronic acid (3.62 g, 15.2 mmol, 1.1 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.02 g, 1.39 mmol, 0.1 eq) and cesium carbonate (9.03 g, 27.8 mmol, 2.0 eq) were dissolved in 1,4-dioxane (40 mL) and water (4 mL) and reacted at 80 ° C under nitrogen protection for 5 hours.
  • reaction solution was filtered, concentrated, and stirred, then purified by normal phase column chromatography (ethyl acetate/petroleum ether, 30% ethyl acetate) to obtain tert-butyl [4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)phenyl]carbamate (2.13 g, 46% yield) as a yellow solid.
  • Step 2 Synthesis of tert-butyl [4-(1,4-dioxaspiro[4.5]decan-8-yl)phenyl]carbamate
  • Step 3 Synthesis of tert-butyl [4-(4-oxocyclohexyl)phenyl]carbamate
  • Step 4 Synthesis of tert-butyl [4-(3-bromo-4-oxocyclohexyl)phenyl]carbamate
  • Step 6 Synthesis of tert-butyl ⁇ 4-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridyl]-3-carboxamido)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl]phenyl ⁇ carbamate
  • 6-Morpholinyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (256 mg, 1.07 mmol, 1.0 eq) was dissolved in acetonitrile (5 mL), and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (298 mg, 1.07 mmol, 1.0 eq), tetramethylchlorouronium hexafluorophosphate (359 mg, 1.21 mmol, 1.2 eq) and N-methylimidazole (263 mg, 3.21 mmol, 3.0 eq) were added. The reaction was carried out at room temperature under nitrogen protection for 12 hours.
  • 1,4-Dioxaspiro[4.5]decane-7-ene-8-trifluoromethanesulfonate (3.4 g, 10.6 mmol, 1.2 eq)
  • tert-butyl[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl]carbamate (2.55 g, 8.85 mmol, 1.0 eq)
  • Step 3 Synthesis of [5-(1,4-dioxaspiro[4.5]n-decane-8-yl)pyridin-2-yl]decane
  • 1,4-Dioxaspirino[4.5]decan-8-one (6.5 g, 41.67 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (40.0 mL). Under nitrogen, a solution of (4-cyanophenyl)magnesium bromide in tetrahydrofuran (75.0 mL, 1.2 eq) was added dropwise at 0°C. The mixture was allowed to react at room temperature for 3 hours. After completion, the reaction was quenched with saturated aqueous ammonium chloride in an ice bath. The mixture was then extracted with water and ethyl acetate.
  • Step 5 Synthesis of 2'-chloro-N-[6-(4-cyanophenyl)-6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • N-methylimidazole (121 mg, 1.48 mmol, 4.0 eq) was added, followed by N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (155 mg, 0.55 mmol, 1.5 eq) in an ice bath.
  • the mixture was reacted at room temperature for 16 hours.
  • preparative separation and purification were performed to obtain 2'-chloro-N-[6-(4-cyanophenyl)-6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (41 mg, yield 20.92%).
  • Step 5 Synthesis of 2'-chloro-N-[6-(4-cyano-2-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • Step 1 Synthesis of 3-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzaldehyde
  • Step 2 Synthesis of 8-[4-(difluoromethyl)-2-methoxyphenyl]-1,4-dioxaspiro[4.5]dec-7-ene
  • Step 3 Synthesis of 8-[4-(difluoromethyl)-2-methoxyphenyl]-1,4-dioxaspiro[4.5]decane
  • Step 5 Synthesis of 6-[4-(difluoromethyl)-2-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine
  • Step 6 Synthesis of 2'-chloro-N- ⁇ 6-[4-(difluoromethyl)-2-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl ⁇ -5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • Step 6 Synthesis of 2'-chloro-N-[6-(4-cyano-3-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • Step 1 Synthesis of 3-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzaldehyde
  • Step 2 Synthesis of 8-[4-(difluoromethyl)-3-methoxyphenyl]-1,4-dioxaspiro[4.5]dec-7-ene
  • Step 3 Synthesis of 8-[4-(difluoromethyl)-3-methoxyphenyl]-1,4-dioxaspiro[4.5]decane
  • Step 5 Synthesis of 6-[4-(difluoromethyl)-3-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine
  • Step 6 Synthesis of 2'-chloro-N- ⁇ 6-[4-(difluoromethyl)-3-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl ⁇ -5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • Step 1 Synthesis of 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine
  • Step 4 Synthesis of 4-[4-(2,2,2-trifluoroethyl)amino]phenylcyclohexane-1-one
  • Step 5 Synthesis of 6- ⁇ 4-[(2,2,2-trifluoroethyl)amino]phenyl ⁇ -4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine
  • Step 6 Synthesis of 2'-chloro-5'-methoxy-6-methyl-N- ⁇ 6-[4-(2,2,2-trifluoroethyl)amino]phenyl ⁇ -4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide
  • N-methyl-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzenesulfonamide (2.4 g, 6.4 mmol, 1.0 eq) and palladium on carbon (480 mg) were dissolved in methanol (5 mL). The mixture was replaced with hydrogen three times and allowed to react at room temperature for 16 hours. After the reaction was complete, the mixture was filtered and the filtrate was concentrated to obtain N-methyl-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzenesulfonamide (2.4 g) as a white solid, which was used directly in the next reaction.
  • N-Methyl-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzenesulfonamide (800 g, 2.57 mmol, 1.0 eq) and p-toluenesulfonic acid hydrate (977 mg, 5.1 mmol, 2.0 eq) were dissolved in acetone (9 mL) and water (3 mL) and reacted at 60°C for 16 hours. After the reaction was complete, the mixture was quenched with sodium bicarbonate solution and extracted with ethyl acetate and water.
  • N,N-Dimethyl-4- ⁇ 1,4-dioxaspiro[4.5]dec-8-yl ⁇ benzenesulfonamide 600 mg, 1.85 mmol, 1.0 eq
  • p-toluenesulfonic acid hydrate 527 mg, 2.77 mmol, 1.5 eq
  • acetone 9 mL
  • water 3 mL
  • the mixture was quenched with sodium bicarbonate solution and extracted with ethyl acetate and water.
  • N,N-Dimethyl-4-(4-oxocyclohexyl)benzenesulfonamide 150 mg, 0.534 mmol, 1.0 eq
  • thiourea 81.3 mg, 1.07 mmol, 2.0 eq
  • iodine 163 mg, 0.641 mmol, 1.2 eq
  • Step 4 Synthesis of 2'-chloro-N- ⁇ 6-[4-(N,N-dimethylsulfamoyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl ⁇ -5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • Step 6 Synthesis of 6- ⁇ 6-[(2,2,2-trifluoroethyl)amino]pyridin-3-yl ⁇ -4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine
  • Step 7 Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-(6-((2,2,2-trifluoroethyl)amino)pyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide
  • N-ethyl-4-(4-oxocyclohexyl)benzenesulfonamide (9.1 g, 32.38 mmol, 1.0 eq), thiourea (4.9 g, 64.79 mmol, 2.0 eq), and iodine (12.3 g, 48.57 mmol, 1.5 eq) were dissolved in ethanol (200 mL) and reacted at 80°C for 2 hours. After the reaction was complete, the reaction mixture was concentrated and then quenched with saturated sodium bicarbonate solution and sodium thiosulfate solution.
  • Step 6 Synthesis of 2'-chloro-5'-methoxy-6-methyl-N- ⁇ 6-[4-(N-ethylaminosulfonyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl ⁇ -[4,4'-bipyridine]-3-carboxamide
  • Step 1 Synthesis of tert-butyl 4-(2,2,2-trifluoroethyl)amino)piperidine-1-carboxylate
  • Step 3 Synthesis of tert-butyl (6-(4-(2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)carbamate
  • tert-butyl (6-(4-(2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)carbamate (600 mg, 1.38 mmol, 1.0 eq) in dichloromethane (8 mL). Slowly add trifluoroacetic acid (1 mL) and react at room temperature for 30 minutes. After the reaction is complete, extract with water and ethyl acetate (100 mL x 3).
  • Step 3 Synthesis of 2'-chloro-5'-methoxy-6-methyl-N- ⁇ 6-[(1-methylazetidin-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl ⁇ -[4,4'-bipyridine]-3-carboxamide
  • 6-Bromo-N-(2,2,2-trifluoroethyl)pyridin-3-amine (427 mg, 1.67 mmol, 1.0 eq), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (489 mg, 1.84 mmol, 1.1 eq), 1,1-bis(diphenylphosphino)diborane iron palladium dichloride (122 mg, 0.167 mmol, 0.1 eq), and potassium carbonate (577 mg, 4.18 mmol, 2.5 eq) were added to 1,4-dioxane (10 ml) and water (2 ml).
  • 6-(1,4-Dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)pyridin-3-amine 328 mg, 1.04 mmol, 1.0 eq
  • palladium on carbon 66 mg
  • the mixture was filtered through celite and the filtrate was concentrated to obtain 6-(1,4-Dioxyaspirino[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)pyridin-3-amine (290 mg, 87.9% yield) as a brown oil.
  • 6-(1,4-Dioxyaspirin[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)pyridin-3-amine 290 mg, 0.92 mmol, 1.0 eq
  • p-toluenesulfonic acid monohydrate 174 mg, 0.92 mmol, 1.0 eq
  • acetone 9 ml
  • water 3 ml
  • ethyl acetate (20 mL) and water (15 mL) were added for extraction.
  • Step 6 Synthesis of 2'-chloro-5'-methoxy-6-methyl-N- ⁇ 6-[5-(2,2,2-trifluoroethyl)amino]pyridin-2-yl ⁇ -4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide
  • 6-Chloropyridine-3-sulfonyl chloride (6 g, 28.29 mol, 1.0 eq), methylamine hydrochloride (2.3 g, 0.034 mol, 1.2 eq), and triethylamine (8.6 g, 0.085 mol, 3.0 eq) were added sequentially to dichloromethane (20 ml) and allowed to react at room temperature for 2 hours.
  • H2O 100 mL
  • ethyl acetate 100 ml x 3
  • the organic phase was concentrated to afford 6-chloro-N-methylpyridine-3-sulfonamide (2.5 g, 43.3% yield) as a brown oil.
  • N-Methyl-6-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyridine-3-sulfonamide 980 mg, 3.16 mmol, 1.0 eq
  • palladium/carbon 67 mg, 0.63 mmol, 0.2 eq
  • Filtration and concentration afforded a crude yellow solid of N-methyl-6-(1,4-dioxaspiro[4.5]dec-8-yl)pyridine-3-sulfonamide (1.0 g), which was used directly in the next reaction.
  • N-Methyl-6-(1,4-dioxaspiro[4.5]dec-8-yl)pyridine-3-sulfonamide (1.0 g, 3.2 mmol, 1.0 eq) and toluenesulfonic acid monohydrate (183 mg, 0.96 mmol, 0.3 eq) were added to acetone/water (6 mL/3 mL) and reacted at 60°C for 2 hours. The pH was adjusted to neutral by adding saturated aqueous sodium bicarbonate. The mixture was extracted with ethyl acetate (15 mL x 2).
  • N-Methyl-6-(4-oxocyclohexyl)pyridine-3-sulfonamide 200 mg, 0.75 mmol, 1.0 eq
  • ethanol 5 ml
  • Iodine (227 mg, 0.89 mmol, 1.2 eq)
  • thiourea 113 mg, 1.49 mmol, 2.0 eq
  • the mixture was allowed to react at 80°C for 2 hours. After completion, the reaction was quenched by addition of saturated sodium thiosulfate solution.
  • Step 7 Synthesis of 2'-chloro-5'-methoxy-6-methyl-N- ⁇ 6-[5-(N-methylsulfamoyl)pyridin-2-yl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl ⁇ -[4,4'-bipyridine]-3-carboxamide
  • 6-(2-Amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-N-methylpyridine-3-sulfonamide 60 mg, 0.18 mmol, 1.0 eq
  • 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid 51 mg, 0.18 mmol, 1.0 eq
  • N,N-dimethylformamide 3 mL
  • N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate 78 mg, 0.27 mmol, 1.5 eq
  • N-methylimidazole 46 mg, 0.54 mmol, 3.0 eq
  • Step 2 Synthesis of N-(8-(4-chlorophenyl)-1,4-dioxaspiro[4.5]decan-8-yl)-2-methylpropane-2-sulfonamide
  • Step 3 Synthesis of 2-methyl-N- ⁇ 8-[4-(2,2,2-trifluoroethyl)amino]phenyl ⁇ -1,4-dioxaspiro[4.5]decan-8-yl)propane-2-sulfonamide
  • N-(8-(4-chlorophenyl)-1,4-dioxaspiro[4.5]decan-8-yl)-2-methylpropane-2-sulfonamide (1.00 g, 2.7 mmol, 1.0 eq) was dissolved in 1,4-dioxane (10 mL), followed by the addition of sodium tert-butoxide (777 mg, 8.09 mmol, 3.0 eq) and methanesulfonic acid (2-di-tert-butylphosphino-3.6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) (230 mg, 0.27 mmol, 0.1 eq).
  • Step 5 Synthesis of 6- ⁇ 4-[(2,2,2-trifluoroethyl)amino]phenyl ⁇ -4,5,6,7-tetrahydrobenzo[d]thiazole-2,6-diamine
  • Step 6 Synthesis of N- ⁇ 6-amino-6-[4-(2,2,2-trifluoroethyl)amino]phenyl ⁇ -4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide
  • the purpose of this test example is to test the effect of compounds on the proliferation of HCT116 BRCA2 -/- cells.
  • DNA polymerase theta is a synthetic lethal target with homologous recombination deficiency and plays an important role in the DNA damage response pathway for double-strand breaks (DSBs).
  • DSBs double-strand breaks
  • HR deficiency homologous recombination-mediated repair
  • MMEJ microhomology-mediated end joining
  • inhibition of Pol ⁇ leads to cell death through the accumulation of toxic RAD51 intermediates and inhibition of the alt-EJ repair pathway. Therefore, by measuring the inhibitory effect of compounds on the proliferation of HCT116 BRCA2 -/- cells, it can be used to screen for Pol ⁇ protein inhibitors.
  • HCT116 BRCA2 knockout cells and screen for single clones.
  • HCT116 BRCA2 -/- and WT cells in logarithmic growth phase were seeded into 96-well plates, with 1000 cells/well (90 ⁇ L) per well, and incubated overnight at 37°C. The next day, 10 ⁇ L of compound was added at various concentrations, with a maximum concentration of 100 ⁇ M.
  • 10 ⁇ L of compound was added at various concentrations, with a maximum concentration of 100 ⁇ M.
  • Nine 10-fold dilutions were performed (final DMSO concentration was 1%), and the cells were incubated at 37°C for 6 days.
  • the old culture medium was aspirated and 110 ⁇ L of culture medium (culture medium to CCK8 at a ratio of 100:10) was added.
  • the cells were incubated at 37°C for 1-4 hours.
  • the absorbance was measured at 450 nM, and the IC50 value was calculated using GraphPad software. Compounds were screened by
  • the compounds of the present invention can inhibit the proliferation of HCT116 BRCA2-/- cells, with IC50 values reaching the nM level.
  • the activity of some compounds is surprisingly significantly better than that of the positive control. This strong inhibitory effect has important therapeutic significance for the treatment of symptoms or diseases related to Pol ⁇ inhibition; while the inhibitory effect on HCT116 WT cells is weak, with excellent selectivity.
  • the purpose of this test case is to test the effect of compounds on the proliferation of DLD1 BRCA2 -/- cells.
  • DLD1 BRCA2 knockout cells were constructed and single clones were screened.
  • DLD1 BRCA2 -/- and WT cells in logarithmic growth phase were seeded into 96-well plates, with 1000 cells/well (90 ⁇ L) per well, and incubated overnight at 37°C. The next day, 10 ⁇ L of compound was added at various concentrations, with a maximum concentration of 100 ⁇ M.
  • 10 ⁇ L of compound was added at various concentrations, with a maximum concentration of 100 ⁇ M.
  • Nine 10-fold dilutions were performed (final DMSO concentration was 1%), and the cells were incubated at 37°C for 6 days.
  • the old culture medium was aspirated and 110 ⁇ L of culture medium (culture medium to CCK8 ratio of 100:10) was added.
  • the cells were incubated at 37°C for 1-4 hours.
  • the absorbance was measured at 450 nM, and the IC50 was calculated using GraphPad software. Compounds were screened by
  • Pol ⁇ consists of a C-terminal family A DNA polymerase and an N-terminal superfamily 2 (SF2) DNA helicase, separated by a long, poorly conserved central domain of unknown function.
  • ART558 binds to an allosteric site within the Pol ⁇ polymerase catalytic domain and inhibits MMEJ in a dose-dependent manner. Therefore, compounds that inhibit Pol ⁇ polymerase function can be used to screen for Pol ⁇ inhibitors.
  • POL ⁇ -polymerase domain protein expression The POL ⁇ -polymerase domain sequence (residues 1819-2590) was cloned into the pET24N vector containing an N-terminal (His) 6x tag and a tobacco etch virus (TEV) protease cleavage site.
  • the constructed vector was transformed into Escherichia coli Rosetta (DE3) and incubated at an OD of 0.6. 1 mM IPTG was added and cultured at 12°C for 16 h. The cells were harvested, ultrasonically disrupted, and the supernatant was centrifuged. The protein was purified by Ni-NTA and dialyzed for determination of protein concentration and purity.
  • Purified POL ⁇ -polymerase domain protein was added to assay buffer (20 mM Tris, pH 7.80, 50 mM KCl, 10 mM MgCl, 1 mM DTT, 0.01% BSA, 0.01% Tween 20) at a protein concentration of 4 nM. Test compounds (an 11-point dilution series) were then added. The enzyme and test compound inhibitor mixture were incubated at room temperature for 15 minutes.
  • the compounds of the present invention have very strong inhibitory effects on the polymerase function of the Pol ⁇ -polymerase domain protein, with IC50 values reaching the nanomolar range. Some compounds have inhibitory effects comparable to or superior to the positive control drug. This strong inhibitory effect has important therapeutic implications for the treatment of conditions or diseases associated with Pol ⁇ inhibition.
  • the compound of the present invention and control compound A were administered by gavage (50 mg/kg), PO solvent: 40% PEG300 + 15% Tween 80 + 45% water, prepared on the day of administration.
  • Blood was collected from the jugular vein 0h before administration and 0.25h, 0.5h, 1h, 2h, 4h, 7h, 24h after administration (gavage), and about 0.02mL of each sample was collected.
  • K2-EDTA was used for anticoagulation and the samples were placed on ice after collection.
  • Plasma sample processing After blood collection, the blood samples were placed on ice and centrifuged within 1 hour to separate the plasma (centrifugation conditions: 4000g, 5 minutes, 2-8°C). Plasma samples were stored in a -80°C refrigerator before analysis.
  • Phoenix WinNonlin7.0 was used to calculate the pharmacokinetic parameters based on the blood drug concentration data at different time points. The experimental results are shown in Table 4 below:
  • mice The pharmacokinetic data of the compounds of the present invention in mice showed that the oral exposure of the tested representative compounds in mice was unexpectedly good compared with the control compound A.

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Abstract

The present invention relates to a heterocyclic compound for inhibiting the activity of a DNA polymerase Theta(Polθ), a stereoisomer, a tautomer, or a pharmaceutically acceptable salt of the compound and a use thereof in preparation of a drug for treating or preventing cancer. Specifically, provided are a compound as shown in formula I or a stereoisomer form and pharmaceutically acceptable salt thereof, wherein the definition of each group is as described in the description. The compound has relatively high Polθ inhibitory activity, so that the compound can be used for treating diseases related to overexpression of Polθ.

Description

Polθ抑制剂及其药物组合物和应用Polθ inhibitors and pharmaceutical compositions and applications thereof 技术领域Technical Field

本发明属于药物化学领域，具体涉及一种Polθ抑制剂或其立体异构体、互变异构体、药学上可接受的盐，并且涉及该化合物的药物组合物及其在治疗癌症中的用途。The present invention belongs to the field of medicinal chemistry, and specifically relates to a Polθ inhibitor or its stereoisomers, tautomers, and pharmaceutically acceptable salts, as well as to a pharmaceutical composition of the compound and its use in treating cancer.

背景技术Background Art

DNA损伤修复途径包括碱基切除修复(base excision repair，BER)、核苷酸切除修复(nucleotide excision repair,NER)、DNA错配修复(mismatch repair，MMR)以及DNA双链断裂(double-strand break,DSB)修复等。双链DNA损伤修复途径主要包括同源重组(HR)、非同源末端接合(NHEJ)和微同源介导的末端连接(alt-EJ或者MMEJ)三类。同源重组主要依靠同源染色体来指导损伤DNA的校正，而非同源末端接合则直接将断开的两条DNA链连接到一起来实现修复目的。同源重组中的关键蛋白BRCA1和BRCA2是两个重要的抑癌因子，当这两个蛋白缺失时，细胞内同源重组的速率会大大降低且会增加细胞对电离辐射的敏感性，最终导致乳腺癌以及卵巢癌的发病几率升高。另一方面，非同源末端接合这一修复途径的正常运转也尤为重要，相关基因的缺陷会导致毛细血管扩张性运动失调以及范可尼贫血等疾病。MMEJ的修复是由DNA末端侧翼的微同源序列的退火驱动，最初被认为是基因修复的备用途径，随着研究进展，也发现MMEJ并非是一种备份的修复机制。DNA damage repair pathways include base excision repair (BER), nucleotide excision repair (NER), DNA mismatch repair (MMR), and DNA double-strand break (DSB) repair. Double-strand DNA damage repair pathways mainly include homologous recombination (HR), non-homologous end joining (NHEJ), and microhomology-mediated end joining (alt-EJ or MMEJ). Homologous recombination mainly relies on homologous chromosomes to guide the correction of damaged DNA, while non-homologous end joining directly connects the two broken DNA chains together to achieve the purpose of repair. BRCA1 and BRCA2, the key proteins in homologous recombination, are two important tumor suppressor factors. When these two proteins are missing, the rate of homologous recombination in cells will be greatly reduced and the sensitivity of cells to ionizing radiation will increase, ultimately leading to an increased incidence of breast cancer and ovarian cancer. On the other hand, the proper functioning of the repair pathway known as non-homologous end joining (NHEJ) is also crucial. Defects in related genes can lead to diseases such as ataxia telangiectasia and Fanconi anemia. MMEJ repair is driven by the annealing of microhomology sequences flanking the DNA ends and was initially considered a backup pathway for gene repair. However, as research progresses, it has been revealed that MMEJ is not a backup repair mechanism.

DNA聚合酶θ(Polθ或POLQ)是具有同源重组(HR，homologous recombination)缺陷的合成致死型靶点，在双链断裂(DSB)的DNA损伤应答(DDR)途径中具有重要作用。当发生DNA末端切除时，在存在BRCA2的情况下，BRCA2不仅将重组酶RAD51募集到DSB以促进HR，而且还抑制alt-NHEJ等修复途径。当同源重组介导的修复受到损害时(HR缺陷)，如BRCA1或BRCA2突变，Polθ高度表达并引导DSB修复朝向alt-EJ，开启微同源介导的末端连接(MMEJ)的DNA修复过程。在HR缺陷的情况下，Polθ的抑制通过毒性RAD51中间体的积累和alt-EJ修复途径的抑制，导致细胞死亡。DNA polymerase θ (Polθ or POLQ) is a synthetic lethal target of homologous recombination (HR) defects and plays an important role in the DNA damage response (DDR) pathway for double-strand breaks (DSBs). When DNA end resection occurs, in the presence of BRCA2, BRCA2 not only recruits the recombinase RAD51 to the DSB to promote HR but also inhibits repair pathways such as alt-NHEJ. When homologous recombination-mediated repair is impaired (HR deficiency), as in the case of BRCA1 or BRCA2 mutations, Polθ is highly expressed and directs DSB repair toward alt-NHEJ, initiating the DNA repair process of microhomology-mediated end joining (MMEJ). In the context of HR deficiency, inhibition of Polθ leads to cell death through the accumulation of toxic RAD51 intermediates and inhibition of the alt-NHEJ repair pathway.

Polθ仅在少数组织类型中表达，但其在许多癌细胞中高度表达，Polθ能够赋予对癌症疗法的抗性并促进缺乏DNA损伤修复途径的异常细胞的存活。DNA聚合酶θ(Polθ)的主要功能是充当逆转录酶作用，在健康细胞中，Polθ作为RNA介导的DNA修复。而在癌细胞中，Polθ高度表达，并促进并促进癌细胞生长和耐药性。Pol θ is expressed in only a few tissue types but is highly expressed in many cancer cells. Pol θ can confer resistance to cancer therapies and promote the survival of abnormal cells lacking DNA damage repair pathways. The primary function of DNA polymerase θ (Pol θ) is to act as a reverse transcriptase. In healthy cells, Pol θ acts as an RNA-mediated DNA repair enzyme. In cancer cells, Pol θ is highly expressed and promotes cancer cell growth and drug resistance.

美国托马斯杰斐逊大学和南加州大学的研究人员在国际顶尖学术期刊Science子刊Science Advances上发表了题为：Polθreverse transcribes RNA and promotes RNA-templated DNA repair的研究论文，揭示了DNA聚合酶θ(Polθ)对RNA的活性及其如何促进DNA修复和癌细胞增殖，这既挑战了生命科学的基本定律“中心法则”，也提示了Polθ是一个有希望的抗癌药物靶点。Researchers from Thomas Jefferson University and the University of Southern California published a research paper titled "Polθ reverse transcribes RNA and promotes RNA-templated DNA repair" in Science Advances, a subsidiary of the top international academic journal Science. The paper revealed the activity of DNA polymerase θ (Polθ) on RNA and how it promotes DNA repair and cancer cell proliferation. This not only challenges the fundamental law of life science, the "central dogma," but also suggests that Polθ is a promising anti-cancer drug target.

CN114127062A、CN114667167A、WO2020160213、WO2022118210、WO2021123785、WO2023134708等文献公开了DNA聚合酶Theta(Polθ)抑制剂。Documents such as CN114127062A, CN114667167A, WO2020160213, WO2022118210, WO2021123785, and WO2023134708 disclose DNA polymerase Theta (Polθ) inhibitors.

发明内容Summary of the Invention

发明概述SUMMARY OF THE INVENTION

本发明人进行了广泛的研究，发现了下式I所示的化合物具有强烈的Polθ抑制功能，能抑制DLD1 BRCA2-/-细胞和HCT116 BRCA2-/-细胞的增殖，活性强、选择性好、小鼠体内口服吸收良好，克服了现有技术中Polθ抑制剂的不足。本发明提供的杂环化合物对Polθ具有良好的抑制作用。The inventors conducted extensive research and discovered that the compound represented by Formula I below has potent Polθ inhibitory activity, inhibiting the proliferation of DLD1 BRCA2-/- cells and HCT116 BRCA2-/- cells. It exhibits strong activity, good selectivity, and good oral absorption in mice, overcoming the shortcomings of prior Polθ inhibitors. The heterocyclic compounds provided by the present invention exhibit excellent Polθ inhibitory effects.

本发明通过以下技术方案解决了以上的技术问题。The present invention solves the above technical problems through the following technical solutions.

一方面，本发明提供了如式(I)所示的化合物或其立体异构体、、互变异构体、药学上可接受的盐：
In one aspect, the present invention provides a compound represented by formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof:

R1、R2、R3、R4、R5、R6和R7分别独立地选自H、氘、卤素、氨基、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基；R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 are each independently selected from H, deuterium, halogen, amino, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy;

R8选自H、羟基、C1-C6烷氧基、NRaRb；R 8 is selected from H, hydroxy, C 1 -C 6 alkoxy, NR a R b ;

R9为苯基、杂芳基、杂环基、-O(CH2)r-环烷基、-O(CH2)r-杂环烷基、-O(CH2)r-杂芳基，所述苯基、杂芳基、杂环基、-O(CH2)r-环烷基、-O(CH2)r-杂环烷基、-O(CH2)r-杂芳基可任选被1个或多个Rn取代；R 9 is phenyl, heteroaryl, heterocyclyl, -O(CH 2 ) r -cycloalkyl, -O(CH 2 ) r -heterocycloalkyl, -O(CH 2 ) r -heteroaryl, and the phenyl, heteroaryl, heterocyclyl, -O(CH 2 ) r -cycloalkyl, -O(CH 2 ) r -heterocycloalkyl, -O(CH 2 ) r -heteroaryl may be optionally substituted with one or more R n ;

Rn选自H、卤素、氰基、NRaRb、羟基、C1-C6烷基、C3-C6环烷基、C1-C6羟基烷基、C1-C6烷氨基、C1-C6卤代烷氧基、-(CH2)r-O-C1-C6烷基、-C(O)NRaRb、-S(O)2NRaRb、-NH(CH2)rC1-C6卤代烷基、-NH(CH2)rOH、-(CH2)rOC1-C6卤代烷基、C1-C6卤代烷基；R n is selected from H, halogen, cyano, NR a R b , hydroxy, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, C 1 -C 6 hydroxyalkyl, C 1 -C 6 alkylamino, C 1 -C 6 haloalkoxy, -(CH 2 ) r -OC 1 -C 6 alkyl, -C(O)NR a R b , -S(O) 2 NR a R b , -NH(CH 2 ) r C 1 -C 6 haloalkyl, -NH(CH 2 ) r OH, -(CH 2 ) r OC 1 -C 6 haloalkyl, C 1 -C 6 haloalkyl;

Ra、Rb各自独立地选自不存在、氢、C1-C6烷基；R a and R b are each independently selected from absent, hydrogen, and C 1 -C 6 alkyl;

r为0至3的整数。r is an integer from 0 to 3.

作为优选的技术方案，其中所述R9选自可任选被1个或多个Rn取代的苯基、含1至3个选自N、O、S的5-6元杂芳基、含1至3个选自N、O、S的5-6杂环基、-O(CH2)r-C3-C6环烷基、-O(CH2)r-C3-C6杂环烷基、-O(CH2)r-5-6元杂芳基。As a preferred technical solution, R 9 is selected from phenyl which may be optionally substituted by one or more R n , 5-6 membered heteroaryl containing 1 to 3 groups selected from N, O, and S, 5-6 membered heterocyclic group containing 1 to 3 groups selected from N, O, and S, -O(CH 2 ) r -C 3 -C 6 cycloalkyl, -O(CH 2 ) r -C 3 -C 6 heterocycloalkyl, and -O(CH 2 ) r -5-6 membered heteroaryl.

作为优选的技术方案，其中所述R9选自
As a preferred technical solution, wherein said R 9 is selected from

作为优选的技术方案，其中R1选自氢。As a preferred technical solution, wherein R 1 is selected from hydrogen.

作为优选的技术方案，其中R2选自甲基。As a preferred technical solution, R 2 is selected from methyl.

作为优选的技术方案，其中R3选自氢。As a preferred technical solution, R 3 is selected from hydrogen.

作为优选的技术方案，其中R4选自甲氧基。As a preferred technical solution, R 4 is selected from methoxy.

作为优选的技术方案，其中R5选自氢。As a preferred technical solution, R 5 is selected from hydrogen.

作为优选的技术方案，其中R7选自氢。As a preferred technical solution, wherein R 7 is selected from hydrogen.

作为优选的技术方案，本发明所述的化合物或其立体异构体、互变异构体、药学上可接受的盐，其具有结构式(I-1)：
As a preferred technical solution, the compound of the present invention or its stereoisomers, tautomers, and pharmaceutically acceptable salts has the structural formula (I-1):

作为优选的技术方案，其中R6选自卤素、氨基、C1-C6烷基、C1-C6卤代烷基。As a preferred technical solution, R 6 is selected from halogen, amino, C 1 -C 6 alkyl, and C 1 -C 6 haloalkyl.

作为更优选的技术方案，其中R6选自氯、甲基、二氟甲基、氨基。As a more preferred technical solution, R 6 is selected from chlorine, methyl, difluoromethyl, and amino.

作为最优选的技术方案，其中R6选自氯。As the most preferred technical solution, R 6 is selected from chlorine.

作为优选的技术方案，其中R8选自氢、羟基、甲氧基、氨基、氨基烷基。As a preferred technical solution, R 8 is selected from hydrogen, hydroxyl, methoxy, amino, and aminoalkyl.

作为更优选的技术方案，R8选自氢、羟基、甲氧基、氨基、氨基甲基。As a more preferred technical solution, R 8 is selected from hydrogen, hydroxy, methoxy, amino, and aminomethyl.

第二方面，本发明还提供了如下所示化合物或其立体异构体、互变异构体、药学上可接受的盐，其中所述化合物选自：


In a second aspect, the present invention further provides the following compounds or their stereoisomers, tautomers, and pharmaceutically acceptable salts, wherein the compound is selected from:


第三方面，本发明还提供了以上所述化合物、互变异构体及其立体异构体或其药学上可接受的盐，以及一种或多种药学上可接受的载体或赋形剂。In a third aspect, the present invention further provides the above-mentioned compounds, tautomers and stereoisomers thereof or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers or excipients.

第四方面，本发明还提供了所述化合物、互变异构体及其立体异构体或其药学上可接受的盐或组合物在制备用于抑制Polθ过表达的疾病的药物中的用途。In a fourth aspect, the present invention also provides the use of the compound, tautomers and stereoisomers thereof or pharmaceutically acceptable salts or compositions thereof in the preparation of drugs for inhibiting diseases caused by overexpression of Polθ.

作为优选的技术方案，所述疾病为癌症。As a preferred technical solution, the disease is cancer.

作为更优选的技术方案，所述癌症为实体瘤、乳腺癌、肺癌、膀胱癌、食道癌、前列腺癌、胰腺癌、宫颈癌。As a more preferred technical solution, the cancer is solid tumor, breast cancer, lung cancer, bladder cancer, esophageal cancer, prostate cancer, pancreatic cancer, or cervical cancer.

第五方面，本发明还提供了制备在患者中治疗同源重组(HR)缺陷癌症的药物的用途，包括向患者给药治疗有效量的所述化合物或组合物。In a fifth aspect, the present invention also provides a method for preparing a drug for treating homologous recombination (HR)-deficient cancer in a patient, comprising administering a therapeutically effective amount of the compound or composition to the patient.

第六方面，本发明还提供了制备在患者中治疗癌症的药物的用途，包括向患者给药治疗有效量的所述化合物或组合物，其中所述癌症的特征为BRCA基因表达的减少或不存在、BRCA基因的不存在、或BRCA蛋白功能的减少。In a sixth aspect, the present invention also provides the use of preparing a drug for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of the compound or composition, wherein the cancer is characterized by reduced or absent BRCA gene expression, absent BRCA genes, or reduced BRCA protein function.

不受理论束缚，预期本发明的Polθ抑制剂显示某些特性，使它们在某些癌症的治疗治疗中特别有用。例如，在一个实施方案中，本发明的Polθ抑制剂在BRCA1和BRCA2缺陷型原发性和继发性实体瘤中为适当致死性的，包括乳腺、卵巢、前列腺和胰腺。Without being bound by theory, it is expected that the Pol θ inhibitors of the present invention exhibit certain properties that make them particularly useful in the treatment of certain cancers. For example, in one embodiment, the Pol θ inhibitors of the present invention are moderately lethal in BRCA1 and BRCA2 deficient primary and secondary solid tumors, including breast, ovarian, prostate, and pancreatic.

在另一个实施方案中，本发明的Polθ抑制剂在多种原发性和继发性实体瘤中具有适当的致死性，这些实体瘤通过除BRCA缺陷之外的机制为HRD，例如具有启动子超甲基化的那些。在这些没有DSB修复途径可能被完全下调的肿瘤中，Polθ也可以与另一种DDR调节剂如PARP抑制剂、DNA-PK抑制剂、ATR抑制剂、ATM抑制剂、wee1抑制剂或CHK1抑制剂一起给予。In another embodiment, the Pol θ inhibitors of the present invention have moderate lethality in a variety of primary and secondary solid tumors that are HRD by mechanisms other than BRCA deficiency, such as those with promoter hypermethylation. In these tumors where no DSB repair pathways may be completely downregulated, Pol θ can also be administered together with another DDR modulator such as a PARP inhibitor, DNA-PK inhibitor, ATR inhibitor, ATM inhibitor, wee1 inhibitor or CHK1 inhibitor.

在另一个实施方案中，本发明的Polθ抑制剂在原发性和继发性乳腺、卵巢、前列腺和胰腺肿瘤中具有适当的致死性，所述肿瘤保留BRCA1缺陷，但在暴露于或未暴露于PARPi药物之后，对PARPi治疗具有抗性。In another embodiment, Pol θ inhibitors of the invention are moderately lethal in primary and secondary breast, ovarian, prostate, and pancreatic tumors that retain BRCA1 deficiency but are resistant to PARPi treatment with or without exposure to PARPi drugs.

在另一个实施方案中，本发明的Polθ抑制剂适当地增加包括CRR在内的ORR，将延迟PARPi抗性的发作，将增加复发时间和DFS，并且与PARPi治疗方案一起给予时将增加HRD(BRCA1/2缺陷和其他HRD机制)原发性和继发性肿瘤(乳腺、卵巢、前列腺和胰腺)的OS。In another embodiment, the Polθ inhibitors of the present invention modestly increase ORR, including CRR, will delay the onset of PARPi resistance, will increase time to relapse and DFS, and will increase OS in HRD (BRCA1/2 deficiency and other HRD mechanisms) primary and secondary tumors (breast, ovary, prostate, and pancreas) when administered with PARPi treatment regimens.

本发明还提供了如式(II)所示的化合物或其立体异构体、互变异构体、药学上可接受的盐：The present invention also provides a compound represented by formula (II) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof:

其中，R8和R9定义如前面任一项所述。 wherein R 8 and R 9 are as defined above.

本发明还提供了如下任一所示的中间体化合物或其立体异构体、互变异构体、药学上可接受的盐，

The present invention also provides any of the following intermediate compounds or their stereoisomers, tautomers, and pharmaceutically acceptable salts:

本发明人发现，该类化合物是高效的Polθ抑制剂，具有极强的Polθ抑制活性，选择性好、口服吸收好，药代动力学良好，可以用于制备预防和/或治疗Polθ抑制有关的适应症，包括实体瘤、乳腺癌、肺癌、膀胱癌、食道癌、前列腺癌、胰腺癌、宫颈癌等。本发明基于以上发现而得以完成。The present inventors have discovered that these compounds are highly effective Polθ inhibitors with extremely strong Polθ inhibitory activity, good selectivity, good oral absorption, and favorable pharmacokinetics. They can be used to prepare drugs for the prevention and/or treatment of indications related to Polθ inhibition, including solid tumors, breast cancer, lung cancer, bladder cancer, esophageal cancer, prostate cancer, pancreatic cancer, and cervical cancer. The present invention was completed based on the above discovery.

发明详述Detailed Description of the Invention

下面对本发明的各个方面和特点作进一步的描述。Various aspects and features of the present invention are further described below.

本发明所引述的所有文献，它们的全部内容通过引用并入本文，并且如果这些文献所表达的含义与本发明不一致时，以本发明的表述为准。此外，本发明使用的各种术语和短语具有本领域技术人员公知的一般含义，即便如此，本发明仍然希望在此对这些术语和短语作更详尽的说明和解释，提及的术语和短语如有与公知含义不一致的，以本发明所表述的含义为准。下面是本发明所用多种术语的定义，这些定义适用于本申请整个说明书中所用的术语，除非在具体情况中另作说明。All documents cited in the present invention are incorporated herein by reference in their entirety, and if the meanings expressed in these documents are inconsistent with the present invention, the description of the present invention shall prevail. In addition, the various terms and phrases used in the present invention have the general meanings known to those skilled in the art. Even so, the present invention still hopes to provide a more detailed description and explanation of these terms and phrases herein. If the terms and phrases mentioned are inconsistent with the generally known meanings, the meanings expressed in the present invention shall prevail. The following are definitions of various terms used in the present invention, which apply to the terms used throughout the specification of this application, unless otherwise specified in the specific case.

根据本发明的化合物可以以互变异构形式存在，那么本发明包括全部互变异构形式。The compounds according to the present invention may exist in tautomeric forms and the present invention then includes all tautomeric forms.

本发明的化合物具有不对称中心，本发明中含有不对称取代原子的化合物可以被分离成光学活性或消旋形式，本领域技术人员知晓如何制备光学活性形式，比如通过消旋体拆分或者由光学活性的起始原料合成。除非特别说明具体的立体化学或异构体形式，本发明包括所有手性、非对映异构体和消旋体。制备本发明化合物的方法及其中间体属于本发明的一部分。本发明化合物的所有互变异构体也属于是本发明的一部分。The compounds of the present invention possess asymmetric centers. Compounds of the present invention containing asymmetrically substituted atoms can be separated into optically active or racemic forms. Those skilled in the art will appreciate how to prepare optically active forms, such as by racemate resolution or synthesis from optically active starting materials. Unless otherwise indicated with respect to specific stereochemistry or isomeric forms, the present invention encompasses all chiral, diastereoisomer, and racemic forms. Methods for preparing the compounds of the present invention and intermediates thereto are also intended to be included in the present invention. All tautomers of the compounds of the present invention are also intended to be included in the present invention.

如本文所用，术语“可选的”或“任选地”意指随后描述的事件或情况可以发生或可以不发生，并且该描述包括其中事件或情况发生的情况以及其中事件或情况不发生的情况。例如，“可选地取代的烷基”是指烷基可以被取代以及其中烷基未被取代的事件或情况。As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "optionally substituted alkyl" refers to events or circumstances where the alkyl group may be substituted as well as instances where the alkyl group is not substituted.

术语“取代的”是指具有置换主链的一个或多个碳上的氢的取代基的部分。应理解，“取代”或“由...取代”包括隐含的条件，即这样的取代与被取代的原子和取代基的允许化合价一致，并且该取代产生稳定的化合物，例如其不会自发地经历例如通过重排、环化、消除等的转化。如本文所用，术语“取代的”预期包括有机化合物的所有可允许的取代基。在广义方面，可允许的取代基包括有机化合物的非环状和环状、支链和非支链、碳环和杂环、芳族和非芳族取代基。对于适当的有机化合物，允许的取代基可以是一个或多个并且相同或不同。为了本发明的目的，杂原子例如氮可以具有氢取代基和/或本文所述的有机化合物的任何可允许的取代基，其满足杂原子的化合价。取代基可以包括本文所述的任何取代基，例如卤素、羟基、烷基、烷氧基、氨基、氰基、杂芳基、杂环基等。本领域技术人员将理解，如果合适，取代基自身可以被取代。The term "substituted" refers to a moiety having a substituent that replaces a hydrogen on one or more carbons of the main chain. It should be understood that "substituted" or "substituted by..." includes implicit conditions, i.e., such substitution is consistent with the allowed valence of the substituted atom and the substituent, and the substitution produces a stable compound, for example, it will not spontaneously undergo transformations such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is intended to include all permissible substituents of an organic compound. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of an organic compound. For appropriate organic compounds, the permissible substituents can be one or more and the same or different. For the purposes of the present invention, a heteroatom such as nitrogen can have a hydrogen substituent and/or any permissible substituent of an organic compound as described herein that satisfies the valence of the heteroatom. Substituents can include any substituent described herein, such as halogen, hydroxyl, alkyl, alkoxy, amino, cyano, heteroaryl, heterocyclic, etc. It will be understood by those skilled in the art that substituents may themselves be substituted, if appropriate.

如本文所述的，术语“卤素”、“卤代”等表示氟、氯、溴或碘，特别是表示氟、氯、溴，特别优选氟、氯；术语“氰基”是指其中碳原子与氮原子三键合的基团。术语“烷氧基”属于惯用表达，是指通过一个氧原子连接到分子的其余部分的烷基基团，是指包含氧原子的烷基，这类基团的实例包括甲氧基、乙氧基或丙氧基等。As used herein, the terms "halogen," "halo," and the like refer to fluorine, chlorine, bromine, or iodine, and particularly to fluorine, chlorine, and bromine, with fluorine and chlorine being particularly preferred. The term "cyano" refers to a group in which a carbon atom is triple-bonded to a nitrogen atom. The term "alkoxy" is a customary expression and refers to an alkyl group attached to the rest of the molecule via an oxygen atom, and refers to an alkyl group containing an oxygen atom. Examples of such groups include methoxy, ethoxy, or propoxy.

如本文所述的，术语“烷基”是指具有指定数目碳原子数的烷基，其为直链或支链的烷基，并且其可包括其子基团，例如提及“C1-C6烷基”时，其还可以包括C1-C4烷基、C1-C3烷基、C2-C6烷基、C2-C4烷基等表示的子范围的基团，以及具体基团例如甲基、乙基、正丙基、异丙基、正丁基、仲丁基、叔丁基、戊基、己基等。As used herein, the term "alkyl" refers to an alkyl group having a specified number of carbon atoms, which is a linear or branched alkyl group, and which may include subgroups thereof. For example, when referring to a " C1 - C6 alkyl group", it may also include sub-ranges of groups represented by C1 - C4 alkyl, C1 - C3 alkyl, C2 - C6 alkyl, C2 - C4 alkyl, etc., as well as specific groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, etc.

术语“卤代烷基”是指经由卤代键联与分子其余部分连接的如上所定义的烷基。例如，C1-6卤代烷基是指经由卤代键联与分子其余部分连接的具有1至6个碳原子或1至3个碳原子的烷基。优选的卤代烷基包括但不限于-CH2Cl、-CHCl2、-CF3等。The term "haloalkyl" refers to an alkyl group as defined above that is attached to the rest of the molecule via a halo linkage. For example, a C 1-6 haloalkyl group refers to an alkyl group having 1 to 6 carbon atoms or 1 to 3 carbon atoms that is attached to the rest of the molecule via a halo linkage. Preferred haloalkyl groups include, but are not limited to, -CH 2 Cl, -CHCl 2 , -CF 3 , and the like.

如本文所用，单独或与其它术语组合的术语“羟基(hydroxy)”或“羟基(hydroxyl)”意指-OH。As used herein, the term "hydroxy" or "hydroxyl," by itself or in combination with other terms, means -OH.

术语“杂环”、“杂环的”或“杂环基”可互换使用，并且是指被取代和未被取代的3至7元单环基团、7至11元二环基团、以及10至15元三环基团，其在至少一个环中具有至少一个杂原子(O、S或N)，所述含有杂原子的环优选具有1、2或3个选自O、S和N的杂原子。这种含有杂原子的基团的每个环可以含有一个或两个氧或硫原子和/或一至四个氮原子，条件是每个环中的杂原子总数是四个或更少，并且进一步的条件是所述环含有至少一个碳原子。氮和硫原子可任选地被氧化，并且氮原子可任选被季铵化。完成二环和三环基团的稠环可以仅含有碳原子，并且可以是饱和的、部分饱和的或完全不饱和的。杂环基团可以被附接在任何可用的氮或碳原子上。所述杂环基包括杂环烷基等。The terms "heterocycle", "heterocyclic" or "heterocyclyl" are used interchangeably and refer to substituted and unsubstituted 3 to 7 membered monocyclic groups, 7 to 11 membered bicyclic groups and 10 to 15 membered tricyclic groups having at least one heteroatom (O, S or N) in at least one ring, the heteroatom-containing ring preferably having 1, 2 or 3 heteroatoms selected from O, S and N. Each ring of such heteroatom-containing groups may contain one or two oxygen or sulfur atoms and/or one to four nitrogen atoms, provided that the total number of heteroatoms in each ring is four or less, and further provided that the ring contains at least one carbon atom. Nitrogen and sulfur atoms may be optionally oxidized, and nitrogen atoms may be optionally quaternized. The fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated or fully unsaturated. Heterocyclic groups may be attached to any available nitrogen or carbon atom. The heterocyclic radicals include heterocycloalkyl and the like.

术语“杂芳基”是指被取代和未被取代的芳香族5元或6元单环基团、9元或10元二环基团、以及11元至14元三环基团，其在至少一个环中具有至少一个杂原子(O、S或N)，所述含有杂原子的环优选具有1、2或3个选自O、S和N的杂原子。含有杂原子的杂芳基的每个环可以含有一个或两个氧或硫原子和/或一至四个氮原子，条件是每个环中的杂原子总数是四个或更少，并且每个环具有至少一个碳原子。完成二环和三环基团的稠环可以仅含有碳原子，并且可以是饱和的、部分饱和的或不饱和的。氮和硫原子可任选地被氧化，并且氮原子可任选被季铵化。作为二环或三环的杂芳基必须包括至少一个完全芳香族环，但是其他一个或多个稠环可以是芳香族的或非芳香族的。杂芳基可以被附接在任何环的任何可用氮或碳原子上。在化合价允许的情况下，如果所述另一环是环烷基或杂环，则其另外任选地被＝O(氧代)取代。杂芳基的实例包括但不限于：呋喃基、噻吩基、吡咯基、吡唑基、咪唑基、恶唑基、异恶唑基、噻唑基、异噻唑基、1H-四唑基、恶二唑基、三唑基、吡啶基、嘧啶基、吡嗪基、哒嗪基、苯并恶唑基、苯并异恶唑基、苯并噻唑基、苯并呋喃基、苯并噻吩基、苯并三嗪基、酞嗪基、噻蒽、二苯并呋喃基、二苯并噻吩基、苯并咪唑基、吲哚基、异吲哚基、吲唑基、喹啉基、异喹啉基、喹唑啉基、喹喔啉基、嘌呤基、蝶啶基、9H-咔唑基、α-咔啉、中氮茚基、苯并异噻唑基、苯并恶唑基、吡咯并吡啶基、吡唑并嘧啶基、呋喃并吡啶基、嘌呤基、苯并噻二唑基、苯并恶二唑基、苯并三唑基。The term "heteroaryl" refers to substituted and unsubstituted aromatic 5- or 6-membered monocyclic groups, 9- or 10-membered bicyclic groups, and 11- to 14-membered tricyclic groups having at least one heteroatom (O, S, or N) in at least one ring, the heteroatom-containing ring preferably having 1, 2, or 3 heteroatoms selected from O, S, and N. Each ring of the heteroaryl containing heteroatoms can contain one or two oxygen or sulfur atoms and/or one to four nitrogen atoms, provided that the total number of heteroatoms in each ring is four or less, and each ring has at least one carbon atom. The fused rings completing the bicyclic and tricyclic groups can contain only carbon atoms and can be saturated, partially saturated, or unsaturated. The nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen atom may optionally be quaternized. A bicyclic or tricyclic heteroaryl must include at least one fully aromatic ring, but the other one or more fused rings may be aromatic or non-aromatic. The heteroaryl group may be attached to any available nitrogen or carbon atom of any ring. Where valence permits, if the other ring is a cycloalkyl or heterocycle, it is further optionally substituted with =0 (oxo). Examples of heteroaryl include, but are not limited to, furyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, 1H-tetrazolyl, oxadiazolyl, triazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzofuranyl, benzothienyl, benzotriazinyl, phthalazinyl, thianthracene, dibenzofuranyl, benzothiophenyl ... pyrrolopyridinyl, pyrazolopyrimidinyl, furopyridinyl, purinyl, benzothiadiazolyl, benzoxadiazolyl, benzotriazolyl, benzotriazole, benzothiadiazolyl, benzotriazole ...

某些式(I)化合物可以以盐，除了盐的形式，本发明所提供的化合物还存在前药形式。本文所描述的化合物的前药容易地在生理条件下发生化学变化从而转化成本发明的化合物。此外，前体药物可以在体内环境中通过化学或生化方法被转换到本发明的化合物。Certain compounds of formula (I) may be in the form of salts. In addition to salts, the compounds provided herein may also exist in the form of prodrugs. Prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to convert into the compounds of the present invention. Additionally, prodrugs may be converted to the compounds of the present invention by chemical or biochemical methods in vivo.

如本文所述的，术语“疾病”是指所述受试者的一种身体状态，该身体状态与本发明所述疾病有关。例如，本发明所述癌症。本发明的癌症包括对标准治疗，如外科手术、放射治疗、化疗和激素治疗等。As used herein, the term "disease" refers to a physical condition of the subject that is associated with the disease described herein. For example, cancer described herein. Cancer described herein includes standard treatments such as surgery, radiation therapy, chemotherapy, and hormone therapy.

“癌症”或“恶性肿瘤”是指以不受控制的细胞异常增殖为特征的多种疾病中的任何一种，受影响的细胞在局部或通过血流和淋巴系统扩散到其他部位的能力的身体(即转移)以及许多特征结构和/或分子特征中的任何一个。“癌症包括间皮瘤、成神经细胞瘤、直肠癌、结肠癌、家族性腺瘤性息肉病癌和遗传性非息肉病结肠直肠癌、食管癌、唇癌、喉癌、下咽癌、舌癌、唾液腺癌、胃癌、腺癌、甲状腺髓样癌、甲状腺乳头状癌、肾癌、肾实质癌、卵巢癌、宫颈癌、子宫体癌、子宫内膜癌症、绒毛膜癌、胰腺癌、前列腺癌、膀胱癌、睾丸癌、乳腺癌、泌尿系统癌、黑色素瘤、脑瘤、头颈癌、急性淋巴细胞性白血病(ALL)、慢性淋巴细胞性白血病(CLL)、急性髓性白血病(AML)、慢性髓性白血病(CML)、肝细胞癌、胆囊癌、支气管瘤、晚期实体瘤、小细胞肺癌等。"Cancer" or "malignancy" refers to any of a variety of diseases characterized by uncontrolled abnormal proliferation of cells, the ability of affected cells to spread locally or through the bloodstream and lymphatic system to other sites in the body (i.e., metastasis), and any of a number of characteristic structural and/or molecular features. "Cancer includes mesothelioma, neuroblastoma, rectal cancer, colon cancer, familial adenomatous polyposis and hereditary non-polyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine corpus cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, urinary tract cancer, melanoma, brain tumor, head and neck cancer, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hepatocellular carcinoma, gallbladder cancer, bronchial tumor, advanced solid tumors, small cell lung cancer, etc.

本发明中的化合物或含有它的药物组合物可以单位剂量形式给药，给药途径可为肠道或非肠道，如口服、静脉注射、肌肉注射、静脉滴注、皮下注射、鼻腔、口腔粘膜、眼、肺和呼吸道、皮肤、阴道、直肠等。The compound of the present invention or the pharmaceutical composition containing the same can be administered in a unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, intravenous drip, subcutaneous injection, nasal cavity, oral mucosa, eyes, lungs and respiratory tract, skin, vagina, rectum, etc.

给药剂型可以是液体剂型、固体剂型或半固体剂型。液体剂型可以是溶液剂(包括真溶液和胶体溶液)、乳剂(包括o/w型、w/o型和复乳)、混悬剂、注射剂(包括水针剂、粉针剂和输液)、滴眼剂、滴鼻剂、洗剂和搽剂等；固体剂型可以是片剂(包括普通片、肠溶片、含片、分散片、咀嚼片、泡腾片、口腔崩解片)、胶囊剂(包括硬胶囊、软胶囊、肠溶胶囊)、颗粒剂、散剂、微丸、滴丸、栓剂、膜剂、贴片、气(粉)雾剂、喷雾剂等；半固体剂型可以是软膏剂、凝胶剂、糊剂等。The dosage form can be a liquid dosage form, a solid dosage form, or a semisolid dosage form. Liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including o/w, w/o, and multiple emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, micropills, dropping pills, suppositories, films, patches, aerosols (powders), and sprays; semisolid dosage forms can be ointments, gels, pastes, and the like.

为达到用药目的，增强治疗效果，本发明的药物或药物组合物可用任何公知的给药方法给药。To achieve the purpose of medication and enhance the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known method of administration.

本发明的化合物或组合物可单独服用，或与其他治疗药物或对症药物合并使用。当本发明的化合物与其它治疗药物存在协同作用时，应根据实际情况调整它的剂量。The compound or composition of the present invention can be taken alone or in combination with other therapeutic drugs or symptomatic drugs. When the compound of the present invention has a synergistic effect with other therapeutic drugs, its dosage should be adjusted according to the actual situation.

有益技术效果Beneficial technical effects

本发明人发现，本发明中的化合物具有良好的Polθ抑制活性，IC50小于阳性对照，能抑制DLD1BRCA2-/-细胞和HCT116 BRCA2-/-等细胞的增殖，在小鼠体内口服吸收出乎意料的良好，在小鼠体内口服暴露量出乎意料的良好，且选择性较好。本发明提供了一类结构新颖、活性强的Polθ抑制剂化合物，该类化合物在预防和/或治疗Polθ抑制有关的适应症比如BRCA基因表达的减少或不存在、BRCA基因的不存在、或BRCA蛋白功能的减少等具有良好的应用前景。The inventors have discovered that the compounds of the present invention have excellent Polθ inhibitory activity, with an IC50 lower than that of the positive control, can inhibit the proliferation of DLD1BRCA2-/- cells and HCT116 BRCA2-/- cells, and exhibit unexpectedly good oral absorption and exposure in mice, with good selectivity. The present invention provides a class of Polθ inhibitor compounds with novel structures and strong activity. Such compounds have promising application prospects in preventing and/or treating indications related to Polθ inhibition, such as reduced or absent BRCA gene expression, the absence of BRCA genes, or reduced BRCA protein function.

具体实施方式DETAILED DESCRIPTION

下面所列实施例有助于本领域技术人员更好地理解本发明的技术方案，但不以任何方式限制本发明。The following embodiments are intended to help those skilled in the art better understand the technical solutions of the present invention, but are not intended to limit the present invention in any way.

对于以下全部实施例，可使用本领域技术人员已知的标准操作和方法。除非另有说明，所有温度以℃(摄氏度)表示。化合物的结构是通过核磁共振谱(NMR)和/或质谱(MS)来确定的。For all of the following examples, standard procedures and methods known to those skilled in the art can be used. Unless otherwise indicated, all temperatures are expressed in degrees Celsius. The structures of the compounds are determined by nuclear magnetic resonance spectroscopy (NMR) and/or mass spectroscopy (MS).

本发明的化合物的结构由核磁共振(NMR)或/和液-质谱(LC-MS)确定。NMR化学位移(δ)以百万分之比(ppm)为单位。用Bruker avance-400型核磁仪测定了核磁共振，溶剂为氘代二甲基亚砜(DMSO-d6)、氘代甲醇(CD3OD)和氘代氯仿(CDCl3)，内标为四甲基硅烷(TMS)。The structures of the compounds of the present invention were confirmed by nuclear magnetic resonance (NMR) and/or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are expressed in parts per million (ppm). NMR measurements were performed using a Bruker Avance-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol ( CD3OD ), and deuterated chloroform ( CDCl3 ), with tetramethylsilane (TMS) as the internal standard.

液相质谱LC-MS测量液相部分使用ACQUITY UPLC超高压液相色谱，质谱部分使用Xevo G2-S Qtof质谱仪。Liquid phase mass spectrometry LC-MS measurement uses ACQUITY UPLC ultra-high pressure liquid chromatography for the liquid phase part and Xevo G2-S Qtof mass spectrometer for the mass spectrometry part.

本发明示例中的起始材料是已知的并且可以在市场上购买，也可以使用或按照本领域已知的方法合成。The starting materials used in the examples of the present invention are known and can be purchased commercially, or can be synthesized using or according to methods known in the art.

实施例1：2'-氯-5'-甲氧基-6-甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(化合物1)的合成
Example 1: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (Compound 1)

步骤1：2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸甲酯的合成Step 1: Synthesis of methyl 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylate

将4-溴-6-甲基烟酸甲酯(2g,8.7mmol,1.0eq)溶于1,4-二氧六环(25.0mL)中，加入(2-氯-5-甲氧基吡啶-4-基)硼酸(1.63g,8.7mmol,1.0eq)、碳酸铯(8.478g,26.1mmol,3.0eq)、去离子水(5.0mL)和1,1-双(二苯基膦)二荗铁二氯化钯(636.5mg,0.87mmol,0.1eq)，氮气保护下，80℃反应3小时。反应完成后，过滤，经正相柱分离纯化得白色固体2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸甲酯(1.734g,收率68.3％)。LCMS(ESI)[M+H]+:293.0。1H NMR(400MHz,DMSO)δ8.87(s,1H),8.22(s,1H),7.50(s,1H),7.37(s,1H),3.78(s,3H),3.68(s,3H),2.57(s,3H)。Methyl 4-bromo-6-methylnicotinate (2 g, 8.7 mmol, 1.0 eq) was dissolved in 1,4-dioxane (25.0 mL). (2-chloro-5-methoxypyridin-4-yl)boronic acid (1.63 g, 8.7 mmol, 1.0 eq), cesium carbonate (8.478 g, 26.1 mmol, 3.0 eq), deionized water (5.0 mL), and 1,1-bis(diphenylphosphino)diphenylferric palladium chloride (636.5 mg, 0.87 mmol, 0.1 eq) were added. The mixture was reacted at 80°C under nitrogen for 3 hours. After completion of the reaction, the mixture was filtered and purified by normal phase column chromatography to obtain methyl 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylate (1.734 g, 68.3% yield) as a white solid. LCMS(ESI)[M+H] + :293.0. 1 H NMR (400MHz, DMSO) δ8.87(s,1H),8.22(s,1H),7.50(s,1H),7.37(s,1H),3.78(s,3H),3.68(s,3H),2.57(s,3H).

步骤2：2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸的合成Step 2: Synthesis of 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid

将2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸甲酯(1.734g,5.9mmol,1.0eq)溶于无水甲醇(10mL)中，加入氢氧化锂(0.748g,17.8mmol 3.0eq)，60℃条件下反应3小时。反应完全后，加入稀盐酸将调pH至4，乙酸乙酯萃取，浓缩有机相得到白色固体2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(1.586g,收率96.0％)。LCMS(ESI)[M+H]+:279.0。1H NMR(400MHz,DMSO)δ13.08(s,1H),8.88(s,1H),8.21(s,1H),7.45(s,1H),7.31(s,1H),3.78(s,3H),2.55(s,3H)。Methyl 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylate (1.734 g, 5.9 mmol, 1.0 eq) was dissolved in anhydrous methanol (10 mL), and lithium hydroxide (0.748 g, 17.8 mmol, 3.0 eq) was added. The mixture was reacted at 60°C for 3 hours. After completion of the reaction, the pH was adjusted to 4 by adding dilute hydrochloric acid. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to afford 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (1.586 g, 96.0% yield) as a white solid. LCMS (ESI) [M+H] + : 279.0. 1 H NMR (400MHz, DMSO) δ13.08(s,1H),8.88(s,1H),8.21(s,1H),7.45(s,1H),7.31(s,1H),3.78(s,3H),2.55(s,3H).

步骤3：2'-氯-5'-甲氧基-6-甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺的合成Step 3: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide

将2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(120mg,0.43mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(10mL)中，随后加入6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-胺(98.3mg,0.43mmol 1.0eq)，N-甲基咪唑(107mg,1.3mmol 3.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(168mg,0.65mmol,1.5eq)。室温反应4小时。反应完全后，过滤，浓缩，制备分离纯化，得到黄色固体2'-氯-5'-甲氧基-6-甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(130.0mg,收率65.1％)。LCMS(ESI)[M+H]+:491.1570。1H NMR(400MHz,DMSO)δ12.52(s,1H),8.78(s,1H),8.17(s,1H),7.52(s,1H),7.41(s,1H),7.37-7.27(m,4H),7.22(m,J＝5.5,2.6Hz,1H),3.62(s,3H),3.08-2.87(m,2H),2.81-2.65(m,3H),2.58(s,3H),2.07-1.96(m,2H)。Dissolve 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (120 mg, 0.43 mmol, 1.0 eq) in ultra-dry N,N-dimethylformamide (10 mL). Add 6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (98.3 mg, 0.43 mmol, 1.0 eq), N-methylimidazole (107 mg, 1.3 mmol, 3.0 eq), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (168 mg, 0.65 mmol, 1.5 eq). Allow to react at room temperature for 4 hours. After the reaction was complete, the product was filtered, concentrated, and purified using preparative separation to obtain 2'-chloro-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (130.0 mg, 65.1% yield) as a yellow solid. LCMS (ESI) [M+H] + : 491.1570. 1H NMR (400MHz, DMSO) δ12.52(s,1H),8.78(s,1H),8.17(s,1H),7.52(s,1H),7.41(s,1H),7.37-7.27(m,4H),7. 22(m,J=5.5,2.6Hz,1H),3.62(s,3H),3.08-2.87(m,2H),2.81-2.65(m,3H),2.58(s,3H),2.07-1.96(m,2H).

实施例2：2'-氯-N-[6-(6-氟-4-甲基吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物2)的合成
Example 2: Synthesis of 2'-chloro-N-[6-(6-fluoro-4-methylpyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 2)

步骤1：1,4-二氧阿司匹林[4.5]癸-7-烯-8-基三氟甲磺酸酯的合成Step 1: Synthesis of 1,4-dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate

将1,4-二氧阿司匹林[4.5]癸-8-酮(10g,64.0mmol,1.0eq)溶于超干四氢呋喃(100.0mL)中，在-78℃下条件下，缓慢加入双(三甲硅基)氨基锂(21.45g,128.0mmol,2.0eq)，控制滴加时间为1小时，随后缓慢加入三氟甲磺酸干(21.7g,76.8mmol,1.2eq)，在-78℃下反应6小时。反应完成后，加入饱和氯化铵溶液淬灭反应，再加入水和乙酸乙酯萃取分液，收集并浓缩有机相，经正相柱分离纯化得到1,4-二氧阿司匹林[4.5]癸-7-烯-8-基三氟甲磺酸酯(9.0g,收率48.8％)。LCMS(ESI)[M+H]+:289.1。1,4-Dioxyaspirino[4.5]decan-8-one (10 g, 64.0 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (100.0 mL). Lithium bis(trimethylsilyl)amide (21.45 g, 128.0 mmol, 2.0 eq) was slowly added at -78°C over a 1-hour period. Dry trifluoromethanesulfonic acid (21.7 g, 76.8 mmol, 1.2 eq) was then slowly added and allowed to react at -78°C for 6 hours. After completion, the reaction was quenched with saturated ammonium chloride solution, and the mixture was extracted with water and ethyl acetate. The organic phase was collected and concentrated, and purified by normal phase column chromatography to yield 1,4-dioxyaspirino[4.5]decan-7-en-8-yl trifluoromethanesulfonate (9.0 g, 48.8% yield). LCMS (ESI) [M+H] + : 289.1.

步骤2：2-氟-4-甲基-5-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)吡啶的合成Step 2: Synthesis of 2-fluoro-4-methyl-5-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)pyridine

将1,4-二氧阿司匹林[4.5]癸-7-烯-8-基三氟甲磺酸酯(9.29g,32.3mmol,1.0eq)溶于1,4-二氧六环(50mL)中，加入(6-氟-4-甲基吡啶-3-基)硼酸(5g,32.3mmol,1.0eq)、碳酸铯(31.55g,95.7mmol,3.0eq)、去离子水(10mL)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(943.3mg,1.3mmol,0.04eq)，氮气保护下，80℃反应4小时。反应完全后，经正相柱纯化得到白色固体2-氟-4-甲基-5-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)吡啶(6.1g,收率75.9％)。LCMS(ESI)[M+H]+:250.1。1,4-Dioxyaspirin[4.5]dec-7-en-8-yl trifluoromethanesulfonate (9.29 g, 32.3 mmol, 1.0 eq) was dissolved in 1,4-dioxane (50 mL), and (6-fluoro-4-methylpyridin-3-yl)boric acid (5 g, 32.3 mmol, 1.0 eq), cesium carbonate (31.55 g, 95.7 mmol, 3.0 eq), deionized water (10 mL) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (943.3 mg, 1.3 mmol, 0.04 eq) were added. The mixture was reacted at 80 °C for 4 hours under nitrogen protection. After the reaction was complete, the product was purified by normal phase column chromatography to obtain 2-fluoro-4-methyl-5-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)pyridine (6.1 g, yield 75.9%) as a white solid. LCMS (ESI) [M+H] + : 250.1.

步骤3：2-氟-4-甲基-5-(1,4-二氧阿司匹林[4.5]癸-8-基)吡啶的合成Step 3: Synthesis of 2-fluoro-4-methyl-5-(1,4-dioxyaspirino[4.5]dec-8-yl)pyridine

将2-氟-4-甲基-5-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)吡啶(1.0g,4.02mmol,1.0eq)溶甲醇中(10mL)中，加入钯炭(42.6mg,0.40mmol,0.1eq)，随后氢气氛围下60℃反应10小时。反应完全后，过滤，浓缩有机相得白色固体2-氟-4-甲基-5-(1,4-二氧阿司匹林[4.5]癸-8-基)吡啶(820mg,81.3％)。LCMS(ESI)[M+H]+:252.1。2-Fluoro-4-methyl-5-(1,4-dioxaspirino[4.5]dec-7-en-8-yl)pyridine (1.0 g, 4.02 mmol, 1.0 eq) was dissolved in methanol (10 mL), and palladium on carbon (42.6 mg, 0.40 mmol, 0.1 eq) was added. The mixture was then reacted at 60°C under a hydrogen atmosphere for 10 hours. After completion of the reaction, the mixture was filtered and the organic phase was concentrated to afford 2-fluoro-4-methyl-5-(1,4-dioxaspirino[4.5]dec-8-yl)pyridine (820 mg, 81.3%) as a white solid. LCMS (ESI) [M+H] + : 252.1.

步骤4：4-(6-氟-4-甲基吡啶-3-基)环己烷-1-酮的合成Step 4: Synthesis of 4-(6-fluoro-4-methylpyridin-3-yl)cyclohexane-1-one

将2-氟-4-甲基-5-(1,4-二氧阿司匹林[4.5]癸-8-基)吡啶(400mg,1.59mmol,1.0eq)溶于四氢呋喃(6mL)和乙醇(3mL)中，随后加入盐酸(2mL)，室温反应3小时。反应完全后，加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯萃取，浓缩有机相得白色固体4-(6-氟-4-甲基吡啶-3-基)环己烷-1-酮(320mg,80.2％)。LCMS(ESI)[M+H]+:208.0。1HNMR(400MHz,DMSO)δ8.07(s,1H),7.01(s,1H),3.26(m,J＝12.0,3.4Hz,1H),2.62(m,J＝14.3,6.2Hz,2H),2.44(m,J＝1.6Hz,3H),2.31-2.23(m,2H),2.07-1.96(m,2H),1.93(dd,J＝13.0,3.9Hz,2H)。2-Fluoro-4-methyl-5-(1,4-dioxaspirino[4.5]dec-8-yl)pyridine (400 mg, 1.59 mmol, 1.0 eq) was dissolved in tetrahydrofuran (6 mL) and ethanol (3 mL). Hydrochloric acid (2 mL) was then added and the mixture was allowed to react at room temperature for 3 hours. After completion of the reaction, the pH was adjusted to neutral by adding saturated aqueous sodium bicarbonate solution. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to obtain 4-(6-fluoro-4-methylpyridin-3-yl)cyclohexane-1-one (320 mg, 80.2%) as a white solid. LCMS (ESI) [M+H] + : 208.0. 1 HNMR (400MHz, DMSO) δ8.07(s,1H),7.01(s,1H),3.26(m,J＝12.0,3.4Hz,1H),2.62(m,J＝14.3,6.2H z,2H),2.44(m,J＝1.6Hz,3H),2.31-2.23(m,2H),2.07-1.96(m,2H),1.93(dd,J＝13.0,3.9Hz,2H).

步骤5：6-(6-氟-4-甲基吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 5: Synthesis of 6-(6-fluoro-4-methylpyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-(6-氟-4-甲基吡啶-3-基)环己烷-1-酮(120mg,0.58mmol,1.0eq)溶于乙腈(3mL)中，加入偶氮二异丁腈(94.9mg,0.58mmol,1.0eq)，0℃下加入N-溴代丁二酰亚胺(113.4mg,0.64mmol,1.1eq)，随后80℃反应2小时。待反应完全后，冷却至室温后再加入硫脲(66.0mg,0.88mmol,1.5eq)，80℃反应2小时。反应完全后，采用正相柱进行分离纯化，先用乙酸乙酯冲洗除去杂质，再由二氯甲烷和甲醇(4:1)纯化得到棕色固体6-(6-氟-4-甲基吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(90.0mg，收率59.1％)。LCMS(ESI)[M+H]+:264.0。1HNMR(400MHz,DMSO)δ8.09(s,1H),7.00(s,1H),6.68(s,2H),4.11(s,4H),2.74(dd,J＝15.5,5.1Hz,1H),2.39(s,3H),2.06-1.82(m,2H)。Dissolve 4-(6-fluoro-4-methylpyridin-3-yl)cyclohexane-1-one (120 mg, 0.58 mmol, 1.0 eq) in acetonitrile (3 mL), add azobisisobutyronitrile (94.9 mg, 0.58 mmol, 1.0 eq), and then add N-bromosuccinimide (113.4 mg, 0.64 mmol, 1.1 eq) at 0°C. The mixture is then reacted at 80°C for 2 hours. After the reaction is complete, cool to room temperature, add thiourea (66.0 mg, 0.88 mmol, 1.5 eq), and react at 80°C for 2 hours. After completion of the reaction, separation and purification were performed using a normal phase column, first eluting with ethyl acetate to remove impurities, followed by purification with dichloromethane and methanol (4:1) to afford a brown solid of 6-(6-fluoro-4-methylpyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (90.0 mg, 59.1% yield). LCMS (ESI) [M+H] + : 264.0. 1 H NMR (400 MHz, DMSO) δ 8.09 (s, 1H), 7.00 (s, 1H), 6.68 (s, 2H), 4.11 (s, 4H), 2.74 (dd, J=15.5, 5.1 Hz, 1H), 2.39 (s, 3H), 2.06-1.82 (m, 2H).

步骤6：2'-氯-N-[6-(6-氟-4-甲基吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of 2'-chloro-N-[6-(6-fluoro-4-methylpyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将6-(6-氟-4-甲基吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(90.0mg,0.34mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(95.1mg,0.34mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(2mL)中，加入N-甲基咪唑(84.2mg,1.03mmol,3.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(105.8mg,0.41mmol,1.2eq)，随后室温反应2.0小时。待反应完全后，制备得棕色固体2'-氯-N-[6-(6-氟-4-甲基吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(10.0mg，收率5.6％)。LCMS(ESI)[M+H]+:524.1249。1H NMR(400MHz,DMSO)δ8.78(s,1H),8.17(s,1H),8.13(s,1H),7.51(s,1H),7.40(s,1H),7.02(s,1H),3.62(s,3H),3.21(s,1H),2.97-2.78(m,2H),2.74(s,2H),2.58(s,3H),2.40(s,3H),2.15-1.92(m,2H)。6-(6-Fluoro-4-methylpyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (90.0 mg, 0.34 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (95.1 mg, 0.34 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (2 mL), and N-methylimidazole (84.2 mg, 1.03 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (105.8 mg, 0.41 mmol, 1.2 eq) were added, followed by reaction at room temperature for 2.0 hours. After the reaction was complete, a brown solid, 2'-chloro-N-[6-(6-fluoro-4-methylpyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (10.0 mg, 5.6% yield) was obtained. LCMS (ESI) [M+H] + : 524.1249. 1H NMR (400MHz, DMSO) δ8.78(s,1H),8.17(s,1H),8.13(s,1H),7.51(s,1H),7.40(s,1H),7.02(s,1H),3. 62(s,3H),3.21(s,1H),2.97-2.78(m,2H),2.74(s,2H),2.58(s,3H),2.40(s,3H),2.15-1.92(m,2H).

实施例3：2'-氯-N-{6-[2-氟-4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物3)的合成
Example 3: Synthesis of 2'-chloro-N-{6-[2-fluoro-4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 3)

步骤1：8-[2-氟-4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸-7-烯的合成Step 1: Synthesis of 8-[2-fluoro-4-(trifluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]dec-7-ene

将1,4-二氧阿司匹林[4.5]癸-7en-8-基三氟甲磺酸酯(4.5g,15.61mmol,1.0eq)和[2-氟-4-(三氟甲基)苯基]硼酸(3.24g,15.61mmol,1.0eq)溶于1,4-二氧六环(25mL)，随后加入去离子水(8mL)、碳酸铯(15.27g,46.83mmol,3.0eq)和二氯[1,1'-二(二苯基膦)二茂铁]钯(34.20mg,0.468mmol,0.03eq)，80℃反应2.5小时。反应完成后，正向柱分离纯化得到白色固体8-[2-氟-4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸-7-烯(3.58g,收率75.9％)。LCMS(ESI)[M+H]+:303.0。1H NMR(400MHz,DMSO)δ7.63(d,J＝11.1Hz,1H),7.55(d,J＝6.8Hz,2H),5.96–5.92(m,1H),3.93(s,4H),2.53(s,2H),2.40(t,J＝3.2Hz,2H),1.80(t,J＝6.4Hz,2H)。1,4-Dioxyaspirino[4.5]dec-7en-8-yl trifluoromethanesulfonate (4.5 g, 15.61 mmol, 1.0 eq) and [2-fluoro-4-(trifluoromethyl)phenyl]boronic acid (3.24 g, 15.61 mmol, 1.0 eq) were dissolved in 1,4-dioxane (25 mL), followed by the addition of deionized water (8 mL), cesium carbonate (15.27 g, 46.83 mmol, 3.0 eq) and dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (34.20 mg, 0.468 mmol, 0.03 eq), and the reaction was carried out at 80 ° C for 2.5 hours. After completion of the reaction, forward column separation and purification afforded 8-[2-fluoro-4-(trifluoromethyl)phenyl]-1,4-dioxaspirino[4.5]dec-7-ene (3.58 g, 75.9% yield) as a white solid. LCMS (ESI) [M+H] + : 303.0. 1 H NMR (400 MHz, DMSO) δ 7.63 (d, J = 11.1 Hz, 1H), 7.55 (d, J = 6.8 Hz, 2H), 5.96–5.92 (m, 1H), 3.93 (s, 4H), 2.53 (s, 2H), 2.40 (t, J = 3.2 Hz, 2H), 1.80 (t, J = 6.4 Hz, 2H).

步骤2：8-[2-氟-4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸烷的合成Step 2: Synthesis of 8-[2-fluoro-4-(trifluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]decane

将8-[2-氟-4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸-7-烯(400mg,1.32mmol,1.0eq)溶甲醇(3mL)中，加入钯/碳(14.06mg,0.13mmol,0.1eq)，随后氢气氛围下，室温反应3小时。反应完全后，过滤，浓缩有机相得到白色固体粗品8-[2-氟-4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸烷(385.0mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:305.0。1H NMR(400MHz,DMSO)δ7.63–7.50(m,3H),3.89(s,4H),2.95(d,J＝12.6Hz,1H),1.81–1.64(m,8H)。8-[2-Fluoro-4-(trifluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]dec-7-ene (400 mg, 1.32 mmol, 1.0 eq) was dissolved in methanol (3 mL), and palladium/carbon (14.06 mg, 0.13 mmol, 0.1 eq) was added. The mixture was then reacted at room temperature under a hydrogen atmosphere for 3 hours. After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to afford the crude white solid 8-[2-fluoro-4-(trifluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]decane (385.0 mg), which was used directly in the next reaction. LCMS (ESI) [M+H] + : 305.0. 1 H NMR (400MHz, DMSO) δ7.63–7.50 (m, 3H), 3.89 (s, 4H), 2.95 (d, J = 12.6Hz, 1H), 1.81–1.64 (m, 8H).

步骤3：4-[2-氟-4-(三氟甲基)苯基]环己烷-1-酮的合成Step 3: Synthesis of 4-[2-fluoro-4-(trifluoromethyl)phenyl]cyclohexane-1-one

将8-[2-氟-4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸烷(380mg,1.25mmol,1.0eq)溶于四氢呋喃(6mL)和乙醇(3mL)中，随后加入盐酸(2mL)，室温反应3小时。反应完全后，加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯萃取，浓缩有机相得到白色固体粗品4-[2-氟-4-(三氟甲基)苯基]环己烷-1-酮(424mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:261.1。1H NMR(400MHz,DMSO)δ7.67–7.50(m,3H),3.43(m,J＝10.9,3.7Hz,1H),2.64(m,J＝14.2,6.1Hz,2H),2.29(m,J＝14.8,2.1Hz,2H),2.06(m,J＝12.6,6.2,3.1Hz,2H),1.95(m,J＝13.1,4.4Hz,2H)。8-[2-Fluoro-4-(trifluoromethyl)phenyl]-1,4-dioxaspirin[4.5]decane (380 mg, 1.25 mmol, 1.0 eq) was dissolved in tetrahydrofuran (6 mL) and ethanol (3 mL). Hydrochloric acid (2 mL) was then added and the mixture was allowed to react at room temperature for 3 hours. After the reaction was complete, saturated aqueous sodium bicarbonate was added to adjust the pH to neutral. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to yield a white solid crude product, 4-[2-fluoro-4-(trifluoromethyl)phenyl]cyclohexane-1-one (424 mg), which was used directly in the next reaction. LCMS (ESI) [M+H] + : 261.1. 1H NMR (400MHz, DMSO) δ7.67–7.50(m,3H),3.43(m,J＝10.9,3.7Hz,1H),2.64(m,J＝14.2,6.1Hz,2 H), 2.29 (m, J = 14.8, 2.1Hz, 2H), 2.06 (m, J = 12.6, 6.2, 3.1Hz, 2H), 1.95 (m, J = 13.1, 4.4Hz, 2H).

步骤4：6-[2-氟-4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 4: Synthesis of 6-[2-fluoro-4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-[2-氟-4-(三氟甲基)苯基]环己烷-1-酮(290mg,1.11mmol,1.0eq)和偶氮二异丁腈(182.92mg,1.11mmol,1.0eq)溶于乙腈(3mL)中，0℃条件下，加入N-溴代丁二酰亚胺(21.81mg,1.22mmol,1.1eq)，80℃反应2小时，反应结束后，将反应液冷却至室温，随后再加入硫脲(84.49mg,1.11mmol,1.0eq)，80℃条件下继续反应1小时。反应结束后，正相柱纯化，先用乙酸乙酯冲洗除去杂质，再用二氯甲烷和甲醇体系淋洗，当二氯甲烷和甲醇体积比为20:1时，流出目标产物，浓缩得到类白色固体6-[2-氟-4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(242mg,收率76.82％)。LCMS(ESI)[M+H]+:317.0。1H NMR(400MHz,DMSO)δ7.68–7.60(m,2H),7.56(dd,J＝8.1,1.8Hz,1H),6.79(s,2H),4.04–3.94(m,1H),2.81–2.66(m,2H),2.61–2.53(m,2H),2.03–1.94(m,2H)。4-[2-Fluoro-4-(trifluoromethyl)phenyl]cyclohexane-1-one (290 mg, 1.11 mmol, 1.0 eq) and azobisisobutyronitrile (182.92 mg, 1.11 mmol, 1.0 eq) were dissolved in acetonitrile (3 mL). N-bromosuccinimide (21.81 mg, 1.22 mmol, 1.1 eq) was added at 0°C and reacted at 80°C for 2 hours. After the reaction, the reaction solution was cooled to room temperature and thiourea (84.49 mg, 1.11 mmol, 1.0 eq) was added. The reaction was continued at 80°C for 1 hour. After the reaction, the product was purified by normal phase column, first rinsing with ethyl acetate to remove impurities, then eluting with a dichloromethane and methanol system. The desired product eluted when the volume ratio of dichloromethane to methanol was 20:1. Concentration afforded an off-white solid, 6-[2-fluoro-4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (242 mg, 76.82% yield). LCMS (ESI) [M+H] + : 317.0. 1H NMR (400MHz, DMSO) δ7.68–7.60 (m, 2H), 7.56 (dd, J = 8.1, 1.8Hz, 1H), 6.79 (s, 2H ),4.04–3.94(m,1H),2.81–2.66(m,2H),2.61–2.53(m,2H),2.03–1.94(m,2H).

步骤5：2'-氯-N-{6-[2-氟-4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-N-{6-[2-fluoro-4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将6-[2-氟-4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(113.49mg,0.36mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(100mg,0.36mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(3mL)中，随后加入N-甲基咪唑(88.23mg,1.08mmol,3.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(150.7mg,0.54mmol,1.5eq)，室温反应2小时。反应结束后，制备分离纯化得到白色固体2'-氯-N-{6-[2-氟-4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(42.7mg,收率20.62％)。LCMS(ESI)[M+H]+:577.0884。1H NMR(400MHz,DMSO)δ11.92(s,1H),8.78(s,1H),8.17(s,1H),7.66(dd,J＝9.3,5.9Hz,2H),7.58(d,J＝8.1Hz,1H),7.52(s,1H),7.41(s,1H),3.62(s,3H),3.39(d,J＝14.2Hz,1H),2.96(dd,J＝15.9,5.1Hz,1H),2.83(dd,J＝16.5,10.4Hz,1H),2.78–2.68(m,2H),2.58(s,3H),2.16–2.00(m,2H)。6-[2-Fluoro-4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (113.49 mg, 0.36 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (100 mg, 0.36 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (3 mL). N-methylimidazole (88.23 mg, 1.08 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (150.7 mg, 0.54 mmol, 1.5 eq) were then added and reacted at room temperature for 2 hours. After the reaction, preparative separation and purification were performed to obtain 2'-chloro-N-{6-[2-fluoro-4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (42.7 mg, 20.62% yield) as a white solid. LCMS (ESI) [M+H] + : 577.0884. 1H NMR (400MHz, DMSO) δ11.92(s,1H),8.78(s,1H),8.17(s,1H),7.66(dd,J＝9.3,5.9Hz,2H),7.58(d,J＝8.1Hz,1H),7.52(s,1H),7.41(s,1H),3.62 (s,3H),3.39(d,J＝14.2Hz,1H),2.96(dd,J＝15.9,5.1Hz,1H),2.83(dd,J＝16.5,10.4Hz,1H),2.78–2.68(m,2H),2.58(s,3H),2.16–2.00(m,2H).

实施例4：2'-氯-5'-甲氧基-6-甲基-N-[6-(2,4,5-三氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-[4,4'-联吡啶]-3-甲酰胺(化合物4)的合成
Example 4: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-[6-(2,4,5-trifluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-[4,4'-bipyridine]-3-carboxamide (Compound 4)

步骤1：8-(2,4,5-三氟苯基)-1,4-二氧阿司匹林[4.5]癸-7-烯的合成Step 1: Synthesis of 8-(2,4,5-trifluorophenyl)-1,4-dioxyaspirino[4.5]dec-7-ene

将1,4-二氧阿司匹林[4.5]癸-7-en-8-基三氟甲磺酸酯(8.19g,28.4mmol,1.0eq)和(2,4,5-三氟苯基)硼酸(5.0g,28.4mmol,1.0eq)加入到1,4-二氧六环(80mL)中，再加入碳酸铯(27.78g,85.3mmol,3.0eq)和水(20mL)，最后加入[1,1'-双(二苯基膦基)二茂铁]二氯化钯(831mg,1.1mmol,0.04eq)，在氮气保护下，80℃搅拌反应1.5小时。反应完成后，加入乙酸乙酯和水萃取反应液，收集有机层，用无水硫酸钠干燥，浓缩有机层并经正相柱层析分离纯化得到白色固体8-(2,4,5-三氟苯基)-1,4-二氧阿司匹林[4.5]癸-7-烯(6.6g,收率86.0％)。LCMS(ESI)[M+H]+:271。1H NMR(400MHz,MeOD)δ7.21(m,J＝11.4,9.0,7.0Hz,1H),7.10(m,J＝10.5,6.8Hz,1H),5.84(dd,J＝5.0,3.2Hz,1H),3.99(d,J＝1.8Hz,4H),2.60–2.49(m,2H),2.41(q,J＝2.9Hz,2H),1.86(t,J＝6.4Hz,2H)。1,4-Dioxyaspirin[4.5]dec-7-en-8-yl trifluoromethanesulfonate (8.19 g, 28.4 mmol, 1.0 eq) and (2,4,5-trifluorophenyl)boronic acid (5.0 g, 28.4 mmol, 1.0 eq) were added to 1,4-dioxane (80 mL), followed by cesium carbonate (27.78 g, 85.3 mmol, 3.0 eq) and water (20 mL). Finally, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (831 mg, 1.1 mmol, 0.04 eq) was added. The mixture was stirred at 80 °C under nitrogen protection for 1.5 hours. After the reaction was completed, ethyl acetate and water were added to extract the reaction solution. The organic layer was collected and dried over anhydrous sodium sulfate. The organic layer was concentrated and purified by normal phase column chromatography to obtain 8-(2,4,5-trifluorophenyl)-1,4-dioxaspirino[4.5]dec-7-ene (6.6 g, 86.0% yield) as a white solid. LCMS (ESI) [M+H] + : 271. 1H NMR (400MHz, MeOD) δ7.21 (m, J＝11.4, 9.0, 7.0Hz, 1H), 7.10 (m, J＝10.5, 6.8Hz, 1H), 5.84 (dd, J＝5.0, 3.2Hz, 1H), 3.99 (d, J = 1.8Hz, 4H), 2.60–2.49 (m, 2H), 2.41 (q, J = 2.9Hz, 2H), 1.86 (t, J = 6.4Hz, 2H).

步骤2：8-(2,4,5-三氟苯基)-1,4-二氧阿司匹林[4.5]癸烷的合成Step 2: Synthesis of 8-(2,4,5-trifluorophenyl)-1,4-dioxyaspirino[4.5]decane

将8-(2,4,5-三氟苯基)-1,4-二氧阿司匹林[4.5]癸-7-烯(1.0g,3.7mmol,1.0eq)溶于无水甲醇(10mL)和超干四氢呋喃(5mL)中，再加入钯碳(100mg)，氢气保护下，室温反应16小时。反应完全后，将反应液过滤，浓缩滤液得到白色固体粗品8-(2,4,5-三氟苯基)-1,4-二氧阿司匹林[4.5]癸烷(1.1g)。LCMS(ESI)[M+H]+:273。1H NMR(400MHz,DMSO)δ7.50–7.41(m,1H),7.41–7.33(m,1H),3.88(d,J＝2.5Hz,4H),2.82(d t,J＝15.1,7.6Hz,1H),1.82–1.53(m,8H)。8-(2,4,5-trifluorophenyl)-1,4-dioxyaspirino[4.5]dec-7-ene (1.0 g, 3.7 mmol, 1.0 eq) was dissolved in anhydrous methanol (10 mL) and ultra-dry tetrahydrofuran (5 mL). Palladium on carbon (100 mg) was added and the mixture was allowed to react at room temperature for 16 hours under a hydrogen atmosphere. After the reaction was complete, the reaction solution was filtered and the filtrate was concentrated to obtain the crude product 8-(2,4,5-trifluorophenyl)-1,4-dioxyaspirino[4.5]decane (1.1 g) as a white solid. LCMS (ESI) [M+H] + : 273. 1 H NMR (400MHz, DMSO) δ7.50–7.41(m,1H),7.41–7.33(m,1H),3.88(d,J=2.5Hz,4H),2.82(dt,J=15.1,7.6Hz,1H),1.82–1.53(m,8H).

步骤3：4-(2,4,5-三氟苯基)环己烷-1-酮的合成Step 3: Synthesis of 4-(2,4,5-trifluorophenyl)cyclohexane-1-one

将8-(2,4,5-三氟苯基)-1,4-二氧阿司匹林[4.5]癸烷(1.0g,4.39mmol,1.0eq)溶于四氢呋喃(10mL)和无水乙醇(5mL)中，再加入盐酸(5mL)，室温搅拌反应1小时。反应完全后，向反应液中加水(40mL)，随后在冰浴条件下，少量多次加入氢氧化钠固体调节PH＝8～9，加入乙酸乙酯萃取分液，收集有机层，用无水硫酸钠干燥后浓缩有机层，得到棕色固体粗品4-(2,4,5-三氟苯基)环己烷-1-酮(890mg)。LCMS(ESI)[M+H]+:229。1H NMR(400MHz,DMSO)δ7.51(m,J＝12.2,10.4,6.0Hz,2H),2.61(m,J＝14.2,6.1Hz,2H),2.26(m,J＝14.6,2.0Hz,2H),2.05–2.00(m,1H),1.99(s,2H),1.92(m,J＝12.8,4.0Hz,2H)。8-(2,4,5-Trifluorophenyl)-1,4-dioxaspirino[4.5]decane (1.0 g, 4.39 mmol, 1.0 eq) was dissolved in tetrahydrofuran (10 mL) and anhydrous ethanol (5 mL). Hydrochloric acid (5 mL) was added and the mixture was stirred at room temperature for 1 hour. After the reaction was complete, water (40 mL) was added to the reaction solution. Solid sodium hydroxide was then added in small amounts to adjust the pH to 8-9 under an ice bath. Ethyl acetate was added for extraction. The organic layer was collected, dried over anhydrous sodium sulfate, and concentrated to yield crude 4-(2,4,5-trifluorophenyl)cyclohexane-1-one (890 mg) as a brown solid. LCMS (ESI) [M+H] + : 229. 1H NMR (400MHz, DMSO) δ7.51 (m, J＝12.2, 10.4, 6.0Hz, 2H), 2.61 (m, J＝14.2, 6.1Hz, 2H), 2. 26(m,J＝14.6,2.0Hz,2H),2.05–2.00(m,1H),1.99(s,2H),1.92(m,J＝12.8,4.0Hz,2H).

步骤4：2-溴-4-(2,4,5-三氟苯基)环己烷-1-酮的合成Step 4: Synthesis of 2-bromo-4-(2,4,5-trifluorophenyl)cyclohexan-1-one

将4-(2,4,5-三氟苯基)环己烷-1-酮(880mg,3.86mmol,1.0eq)和偶氮二异丁腈(634mg,3.86mmol,1.0eq)加入到四氯化碳(15mL)中，然后在冰浴条件下，少量多次加入N-溴代琥珀酰亚胺(756mg,4.25mmol,1.1eq)，80℃搅拌反应16小时。反应完全后，直接将反应液浓缩，得到黄色油状物2-溴-4-(2,4,5-三氟苯基)环己烷-1-酮粗品(1.2g)，直接用于下一步反应。4-(2,4,5-trifluorophenyl)cyclohexane-1-one (880 mg, 3.86 mmol, 1.0 eq) and azobisisobutyronitrile (634 mg, 3.86 mmol, 1.0 eq) were added to carbon tetrachloride (15 mL). N-bromosuccinimide (756 mg, 4.25 mmol, 1.1 eq) was then added in small portions in an ice bath. The mixture was stirred at 80°C for 16 hours. After the reaction was complete, the reaction solution was concentrated to obtain crude 2-bromo-4-(2,4,5-trifluorophenyl)cyclohexane-1-one (1.2 g) as a yellow oil, which was used directly in the next reaction.

步骤5：6-(2,4,5-三氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 5: Synthesis of 6-(2,4,5-trifluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

2-溴-4-(2,4,5-三氟苯基)环己烷-1-酮(1.2g,3.92mmol,1.0eq)溶于乙腈(10mL)中，再加入硫脲(358mg,4.7mmol,1.2eq)，80℃搅拌2小时。反应完成后，直接将反应液浓缩，正相柱层析分离纯化得到棕色固体6-(2,4,5-三氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(500mg,收率45.6％)。LCMS(ESI)[M+H]+:285。2-Bromo-4-(2,4,5-trifluorophenyl)cyclohexan-1-one (1.2 g, 3.92 mmol, 1.0 eq) was dissolved in acetonitrile (10 mL), and thiourea (358 mg, 4.7 mmol, 1.2 eq) was added. The mixture was stirred at 80°C for 2 hours. After completion of the reaction, the reaction solution was concentrated and purified by normal phase column chromatography to obtain 6-(2,4,5-trifluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (500 mg, 45.6% yield) as a brown solid. LCMS (ESI) [M+H] + : 285.

步骤6：2'-氯-5'-甲氧基-6-甲基-N-[6-(2,4,5-三氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-[6-(2,4,5-trifluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-[4,4'-bipyridine]-3-carboxamide

将化合物6-(2,4,5-三氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(240mg,0.85mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(235mg,0.85mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(3.5mL)中，再加入N-甲基咪唑(208mg,2.54mmol,3.0eq)，最后在冰浴条件下，加入四甲基氯代脲六氟磷酸酯(356mg,1.27mmol,1.5eq)，常温搅拌反应1.5小时。反应完成后，制备得到白色固体2'-氯-5'-甲氧基-6-甲基-N-[6-(2,4,5-三氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-[4,4'-联吡啶]-3-甲酰胺(12.3mg,收率2.7％)。LCMS(ESI)[M+H]+:545.1237。1H NMR(400MHz,DMSO)δ12.52(s,1H),8.77(s,1H),8.17(s,1H),7.63–7.48(m,3H),7.41(s,1H),3.61(s,3H),3.25(d,J＝12.0Hz,1H),2.90(dd,J＝16.0,5.2Hz,1H),2.84–2.78(m,1H),2.75(d,J＝11.0Hz,2H),2.58(s,3H),2.12–1.94(m,2H)。Compound 6-(2,4,5-trifluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (240 mg, 0.85 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (235 mg, 0.85 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (3.5 mL), and N-methylimidazole (208 mg, 2.54 mmol, 3.0 eq) was added. Finally, tetramethylchlorouronium hexafluorophosphate (356 mg, 1.27 mmol, 1.5 eq) was added in an ice bath and the reaction was stirred at room temperature for 1.5 hours. After completion of the reaction, 2'-chloro-5'-methoxy-6-methyl-N-[6-(2,4,5-trifluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-[4,4'-bipyridine]-3-carboxamide (12.3 mg, 2.7% yield) was obtained as a white solid. LCMS (ESI) [M+H] + : 545.1237. 1H NMR (400MHz, DMSO) δ12.52(s,1H),8.77(s,1H),8.17(s,1H),7.63–7.48(m,3H),7.41(s,1H),3.61(s,3H),3.25(d,J＝ 12.0Hz, 1H), 2.90 (dd, J＝16.0, 5.2Hz, 1H), 2.84–2.78 (m, 1H), 2.75 (d, J＝11.0Hz, 2H), 2.58 (s, 3H), 2.12–1.94 (m, 2H).

实施例5：2'-氯-N-(6-(4-氰基-3-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物5)的合成
Example 5: Synthesis of 2'-chloro-N-(6-(4-cyano-3-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 5)

步骤1：2-氟-4-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)苄腈的合成Step 1: Synthesis of 2-fluoro-4-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)benzonitrile

将1,4-二氧阿司匹林[4.5]癸-7-烯-8-基三氟甲磺酸酯(2.0g,6.94mmol,1.0eq)溶于1,4-二氧六环(10mL)中，加入(4-氰基-3-氟苯基)硼酸(1.01g,6.94mol,1.0eq)、碳酸铯(6.79g,20.8mmol,3.0eq)、去离子水(2mL)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(253.8mg,0.35mmol,0.05eq)，氮气保护下，80℃反应4小时。反应完全后，经正相柱纯化得到白色固体2-氟-4-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)苄腈(1.516g,收率84.3％)。LCMS(ESI)[M+H]+:260。1HNMR(400MHz,DMSO)δ7.84(dd,J＝8.2,7.2Hz,1H),7.57(dd,J＝11.7,1.7Hz,1H),7.47(dd,J＝8.3,1.7Hz,1H),6.36(m,J＝4.0,2.0Hz,1H),3.95(s,2H),2.55(m,J＝6.2,1.9Hz,2H),2.41(dt,J＝4.5,2.4Hz,2H),2.36(t,J＝7.1Hz,1H),1.93(t,J＝7.0Hz,1H),1.81(t,J＝6.5Hz,2H)。1,4-Dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (2.0 g, 6.94 mmol, 1.0 eq) was dissolved in 1,4-dioxane (10 mL). (4-Cyano-3-fluorophenyl)boronic acid (1.01 g, 6.94 mol, 1.0 eq), cesium carbonate (6.79 g, 20.8 mmol, 3.0 eq), deionized water (2 mL), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (253.8 mg, 0.35 mmol, 0.05 eq) were added. The mixture was reacted at 80°C under nitrogen for 4 hours. After completion of the reaction, the product was purified by normal phase column chromatography to obtain 2-fluoro-4-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)benzonitrile (1.516 g, 84.3% yield) as a white solid. LCMS(ESI)[M+H] + :260. 1 HNMR (400MHz, DMSO) δ7.84 (dd, J＝8.2, 7.2Hz, 1H), 7.57 (dd, J＝11.7, 1.7Hz, 1H), 7.47 (dd, J＝8.3, 1.7Hz, 1H), 6.36 (m, J＝4.0, 2.0Hz, 1H), 3.95(s,2H),2.55(m,J＝6.2,1.9Hz,2H),2.41(dt,J＝4.5,2.4Hz,2H),2.36(t,J＝7.1Hz,1H),1.93(t,J＝7.0Hz,1H),1.81(t,J＝6.5Hz,2H).

步骤2：2-氟-4-(1,4-二氧阿司匹林[4.5]癸-8-基)苄腈的合成Step 2: Synthesis of 2-fluoro-4-(1,4-dioxyaspirino[4.5]dec-8-yl)benzonitrile

将2-氟-4-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)苄腈(1.0g,3.86mmol,1.0eq)溶甲醇中(5mL)中，加入钯炭(81.8mg,0.772mmol,0.2eq)，随后氢气氛围下，室温反应8小时。待反应完全后，直接将反应液过滤，滤液浓缩后得到粗品白色固体2-氟-4-(1,4-二氧阿司匹林[4.5]癸-8-基)苄腈(820mg,收率89.3％)。LCMS(ESI)[M+H]+:262。2-Fluoro-4-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)benzonitrile (1.0 g, 3.86 mmol, 1.0 eq) was dissolved in methanol (5 mL), and palladium on carbon (81.8 mg, 0.772 mmol, 0.2 eq) was added. The mixture was then reacted at room temperature under a hydrogen atmosphere for 8 hours. After the reaction was complete, the reaction solution was filtered and the filtrate was concentrated to obtain a crude white solid of 2-fluoro-4-(1,4-dioxyaspirino[4.5]dec-8-yl)benzonitrile (820 mg, 89.3% yield). LCMS (ESI) [M+H] + : 262.

步骤3：2-氟-4-(4-氧代环己基)苄腈的合成Step 3: Synthesis of 2-fluoro-4-(4-oxocyclohexyl)benzonitrile

将2-氟-4-甲基-5-(1,4-二氧阿司匹林[4.5]癸-8-基)吡啶(500mg,1.92mmol,1.0eq)溶于四氢呋喃(6mL)和乙醇(3mL)中，随后加入盐酸(2mL)，室温反应3小时。反应完全后，加入饱和碳酸氢钠水溶液调节pH至中性，用乙酸乙酯萃取，收集并浓缩有机相得白色固体2-氟-4-(4-氧代环己基)苄腈(301mg,收率72.4％)。LCMS(ESI)[M+H]+:218。2-Fluoro-4-methyl-5-(1,4-dioxaspirino[4.5]dec-8-yl)pyridine (500 mg, 1.92 mmol, 1.0 eq) was dissolved in tetrahydrofuran (6 mL) and ethanol (3 mL). Hydrochloric acid (2 mL) was then added and the mixture was allowed to react at room temperature for 3 hours. After completion of the reaction, saturated aqueous sodium bicarbonate solution was added to adjust the pH to neutral. The mixture was extracted with ethyl acetate, and the organic phase was collected and concentrated to yield 2-fluoro-4-(4-oxocyclohexyl)benzonitrile (301 mg, 72.4% yield) as a white solid. LCMS (ESI) [M+H] + : 218.

步骤4：4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-2-氟苯甲腈的合成Step 4: Synthesis of 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-2-fluorobenzonitrile

将2-氟-4-(4-氧代环己基)苄腈(300mg,1.37mmol,1.0eq)溶于乙腈(10mL)中，加入偶氮二异丁腈(224.6mg,1.37mmol,1.0eq)，0℃条件下，加入N-溴代丁二酰亚胺(268.2mg,1.5mmol,1.1eq)，随后80℃反应2小时。待反应完全后，冷却至室温后，再加入硫脲(156.2mg,2.05mmol,1.5eq)，80℃反应2小时。反应完全后，采用正相柱进行分离纯化，先用乙酸乙酯冲洗除去杂质，再由二氯甲烷和甲醇(4:1)纯化得到棕色固体4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-2-氟苯甲腈(215mg,收率57.5％)。LCMS(ESI)[M+H]+:274。1HNMR(400MHz,DMSO)δ7.87(t,J＝7.5Hz,1H),7.54(dd,J＝11.0,1.5Hz,1H),7.39(dd,J＝8.1,1.5Hz,1H),7.02(d,J＝17.9Hz,2H),3.55(m,J＝6.1Hz,1H),3.12(d,J＝11.9Hz,2H),2.69-2.51(m,2H),1.95(q,J＝8.6Hz,2H)。2-Fluoro-4-(4-oxocyclohexyl)benzonitrile (300 mg, 1.37 mmol, 1.0 eq) was dissolved in acetonitrile (10 mL), and azobisisobutyronitrile (224.6 mg, 1.37 mmol, 1.0 eq) was added. N-bromosuccinimide (268.2 mg, 1.5 mmol, 1.1 eq) was then added at 0°C, and the mixture was allowed to react at 80°C for 2 hours. After the reaction was complete, the mixture was cooled to room temperature, and thiourea (156.2 mg, 2.05 mmol, 1.5 eq) was added, followed by a reaction at 80°C for 2 hours. After the reaction was complete, the mixture was separated and purified using a normal phase column. The impurities were first removed by rinsing with ethyl acetate, and then purified using dichloromethane and methanol (4:1) to obtain 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-2-fluorobenzonitrile (215 mg, 57.5% yield) as a brown solid. LCMS(ESI)[M+H] + :274. 1 HNMR (400MHz, DMSO) δ7.87(t,J＝7.5Hz,1H),7.54(dd,J＝11.0,1.5Hz,1H),7.39(dd,J＝8.1,1.5Hz,1H),7.02 (d, J = 17.9 Hz, 2H), 3.55 (m, J = 6.1 Hz, 1H), 3.12 (d, J = 11.9 Hz, 2H), 2.69-2.51 (m, 2H), 1.95 (q, J = 8.6 Hz, 2H).

步骤5：2'-氯-N-(6-(4-氰基-3-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-N-(6-(4-cyano-3-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-2-氟苯甲腈(200.0mg,0.73mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(204mg,0.73mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(3mL)中，随后加入N-甲基咪唑(180mg,2.2mmol,3.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(226mg,0.88mmol,1.2eq)，室温反应3.0小时。反应完全后，制备分离纯化得到棕色固体2'-氯-N-(6-(4-氰基-3-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(31.0mg,收率7.9％)。LCMS(ESI)[M+H]+:534.0972。1HNMR(400MHz,DMSO)δ12.52(s,1H),8.77(s,1H),8.17(s,1H),7.89(t,J＝7.5Hz,1H),7.61-7.50(m,2H),7.47-7.39(m,2H),3.61(s,3H),3.19(d,J＝6.8Hz,1H),2.95(dd,J＝15.8,5.0Hz,1H),2.85–2.74(m,1H),2.72(s,2H),2.58(s,3H),2.04(q,J＝8.1Hz,2H)。4-(2-Amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-2-fluorobenzonitrile (200.0 mg, 0.73 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (204 mg, 0.73 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (3 mL), followed by the addition of N-methylimidazole (180 mg, 2.2 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (226 mg, 0.88 mmol, 1.2 eq) and the reaction was carried out at room temperature for 3.0 hours. After the reaction was complete, preparative separation and purification were performed to obtain a brown solid, 2'-chloro-N-(6-(4-cyano-3-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (31.0 mg, 7.9% yield). LCMS (ESI) [M+H] + : 534.0972. 1 HNMR (400MHz, DMSO) δ12.52(s,1H),8.77(s,1H),8.17(s,1H),7.89(t,J＝7.5Hz,1H),7.61-7.50(m,2H),7.47-7.39(m,2H),3.61 (s,3H),3.19(d,J＝6.8Hz,1H),2.95(dd,J＝15.8,5.0Hz,1H),2.85–2.74(m,1H),2.72(s,2H),2.58(s,3H),2.04(q,J＝8.1Hz,2H).

实施例6：2'-氯-N-(6-(3-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物6)的合成
Example 6: Synthesis of 2'-chloro-N-(6-(3-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 6)

步骤1：6-(3-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 1: Synthesis of 6-(3-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-(3-氟苯基)环己烷-1-酮(1.0g,5.20mmol,1.0eq)溶于乙腈(8mL)中，加入2,2'-偶氮二异丁腈(853mg,5.20mmol,1.0eq)，随后在冰水浴下，加入N-溴代丁二酰亚胺(1.02g,5.72mmol,1.1eq)，80℃反应1.5小时。反应完成后，再加入硫脲(396mg,5.20mmol,1.1eq)，80℃继续反应1小时。反应结束后，将反应液冷却，有固体析出，过滤得到类白色固体粗品6-(3-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(402mg)。LCMS(ESI)[M+H]+:249。1H NMR(400MHz,DMSO)δ9.23(s,2H),7.37(q,J＝7.6Hz,1H),7.16(t,J＝7.3Hz,2H),7.06(td,J＝8.6,2.5Hz,1H),6.22(s,6H),3.04(s,1H),2.84–2.75(m,1H),2.65–2.54(m,4H),1.98(s,2H),1.66(s,1H)。4-(3-Fluorophenyl)cyclohexane-1-one (1.0 g, 5.20 mmol, 1.0 eq) was dissolved in acetonitrile (8 mL), and 2,2'-azobisisobutyronitrile (853 mg, 5.20 mmol, 1.0 eq) was added. N-bromosuccinimide (1.02 g, 5.72 mmol, 1.1 eq) was then added in an ice-water bath. The mixture was allowed to react at 80°C for 1.5 hours. After completion of the reaction, thiourea (396 mg, 5.20 mmol, 1.1 eq) was added, and the reaction was continued at 80°C for 1 hour. After the reaction, the reaction solution was cooled, and a solid precipitated. Filtering afforded the crude product, 6-(3-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (402 mg), as an off-white solid. LCMS (ESI) [M+H] + : 249. 1H NMR (400MHz, DMSO) δ9.23 (s, 2H), 7.37 (q, J = 7.6Hz, 1H), 7.16 (t, J = 7.3Hz, 2H), 7.06 (td, J = 8.6, 2.5 Hz,1H),6.22(s,6H),3.04(s,1H),2.84–2.75(m,1H),2.65–2.54(m,4H),1.98(s,2H),1.66(s,1H).

步骤2：2'-氯-N-[6-(3-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 2: Synthesis of 2'-chloro-N-[6-(3-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将6-(3-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(107mg,0.43mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(120mg,0.43mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(3mL)中，加入N-甲基咪唑(106mg,1.29mmol,3.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(181.0mg,0.645mmol,1.5eq)，随后室温反应2.0小时。反应完全后，制备得黄色固体2'-氯-N-[6-(3-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(44.2mg,收率20.18％)。LCMS(ESI)[M+H]+:509.1159。1HNMR(400MHz,DMSO)δ12.54(s,1H),8.81(s,1H),8.18(s,1H),7.53(s,1H),7.47(s,1H),7.36(q,J＝7.5Hz,1H),7.22–7.16(m,2H),7.08–7.01(m,1H),3.62(s,3H),3.11–3.02(m,1H),2.93(dd,J＝15.7,5.0Hz,1H),2.81–2.66(m,3H),2.60(s,3H),2.03(t,J＝6.9Hz,2H)。6-(3-Fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (107 mg, 0.43 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (120 mg, 0.43 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (3 mL), and N-methylimidazole (106 mg, 1.29 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (181.0 mg, 0.645 mmol, 1.5 eq) were added, followed by reaction at room temperature for 2.0 hours. After the reaction was complete, a yellow solid, 2'-chloro-N-[6-(3-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (44.2 mg, yield 20.18%), was obtained. LCMS (ESI) [M+H] + : 509.1159. 1 HNMR (400MHz, DMSO) δ12.54(s,1H),8.81(s,1H),8.18(s,1H),7.53(s,1H),7.47(s,1H),7.36(q,J＝7.5Hz,1H),7.22–7.16(m,2H),7. 08–7.01(m,1H),3.62(s,3H),3.11–3.02(m,1H),2.93(dd,J＝15.7,5.0Hz,1H),2.81–2.66(m,3H),2.60(s,3H),2.03(t,J＝6.9Hz,2H).

实施例7：2'-氯-N-[6-(4-羟基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物7)的合成
Example 7: Synthesis of 2'-chloro-N-[6-(4-hydroxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 7)

步骤1：4-(4-氧代环己基)乙酸苯酯的合成Step 1: Synthesis of 4-(4-oxocyclohexyl)phenyl acetate

将4-(4-羟基苯基)环己烷-1-酮(1.0g,5.26mmol,1.0eq)溶于超干二氯甲烷(50.0mL)中，缓慢滴加乙酰氯(412.9mg,5.26mmol,1.0eq)，常温下反应4小时。反应完成后，加入水和乙酸乙酯萃取分液，合并有机相，浓缩得到粗品4-(4-氧代环己基)乙酸苯酯(880mg,收率72.1％)，直接用于下一步反应。LCMS(ESI)[M+H]+:233.1。Dissolve 4-(4-hydroxyphenyl)cyclohexane-1-one (1.0 g, 5.26 mmol, 1.0 eq) in ultra-dry dichloromethane (50.0 mL). Slowly add acetyl chloride (412.9 mg, 5.26 mmol, 1.0 eq) dropwise. Allow to react at room temperature for 4 hours. After completion, extract with water and ethyl acetate. The organic phases are combined and concentrated to yield crude 4-(4-oxocyclohexyl)phenyl acetate (880 mg, 72.1% yield), which is used directly in the next step. LCMS (ESI) [M+H] + : 233.1.

步骤2：4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)乙酸苯酯的合成Step 2: Synthesis of phenyl 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)acetate

将4-(4-氧代环己基)乙酸苯酯(800mg,3.45mmol,1.0eq)溶于超干乙腈(5mL)中，随后加入偶氮二异丁腈(565.8mg,3.45mmol,1.0eq)，80℃条件下反应1小时。将反应液冷却至室温，再加入硫脲(393mg,5.17mmol,1.5eq)，80℃条件下继续反应1小时。反应结束后，采用正相柱分离纯化得到黄色油状物4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)乙酸苯酯(230mg,收率23.1％)。LCMS(ESI)[M+H]+:289.09。4-(4-Oxocyclohexyl)phenyl acetate (800 mg, 3.45 mmol, 1.0 eq) was dissolved in ultra-dry acetonitrile (5 mL), followed by the addition of azobisisobutyronitrile (565.8 mg, 3.45 mmol, 1.0 eq). The reaction was allowed to react at 80°C for 1 hour. The reaction solution was cooled to room temperature, and thiourea (393 mg, 5.17 mmol, 1.5 eq) was added. The reaction was continued at 80°C for 1 hour. After completion of the reaction, 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)phenyl acetate was isolated and purified using a normal phase column to obtain a yellow oil (230 mg, 23.1% yield). LCMS (ESI) [M+H] + : 289.09.

步骤3：4-[2-(2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺基)-4,5,6,7-四氢苯并[d]噻唑-6-基]乙酸苯酯的合成Step 3: Synthesis of phenyl 4-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridyl]-3-carboxamido)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl]acetate

将4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)乙酸苯酯(200mg,0.69mmol 1.0eq)溶于超干N,N-二甲基甲酰胺(4mL)中，依次加入2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(212mg,0.76mmol,1.1eq)、N-甲基咪唑(172mg,2.09mmol,3.0eq)和N,N,N′,N′-四甲基氯甲脒六氟磷酸盐(214mg,0.83mmol,1.2eq)，室温下反应2.0小时。反应结束后，将反应液过滤，滤液浓缩，并经正相纯化得到棕黄色油状物4-[2-(2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺基)-4,5,6,7-四氢苯并[d]噻唑-6-基)乙酸苯酯(90.0mg,收率18.4％)。LCMS(ESI)[M+H]+:549.1。1H NMR(400MHz,CDCl3)δ8.85(s,1H),8.03(s,1H),8.01(s,2H),7.24-7.20(m,2H),7.06-7.01(m,2H),3.72(s,3H),3.06-2.97(m,2H),2.76(s,2H),2.70(s,3H),2.29(s,3H),2.08-1.77(m,2H)。4-(2-Amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)phenyl acetate (200 mg, 0.69 mmol, 1.0 eq) was dissolved in ultra-dry N,N-dimethylformamide (4 mL), and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (212 mg, 0.76 mmol, 1.1 eq), N-methylimidazole (172 mg, 2.09 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (214 mg, 0.83 mmol, 1.2 eq) were added in sequence and reacted at room temperature for 2.0 hours. After the reaction, the reaction mixture was filtered, the filtrate was concentrated, and the product was purified by normal phase chromatography to obtain 4-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridyl]-3-carboxamido)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)phenyl acetate (90.0 mg, 18.4% yield) as a brown oil. LCMS (ESI) [M+H] + : 549.1. 1H NMR(400MHz, CDCl3)δ8.85(s,1H),8.03(s,1H),8.01(s,2H),7.24-7.20(m,2H),7.06-7.01(m, 2H),3.72(s,3H),3.06-2.97(m,2H),2.76(s,2H),2.70(s,3H),2.29(s,3H),2.08-1.77(m,2H).

步骤4：2'-氯-N-[6-(4-羟基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 4: Synthesis of 2'-chloro-N-[6-(4-hydroxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将4-[2-(2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺基)-4,5,6,7-四氢苯并[d]噻唑-6-基)乙酸苯酯(70mg,0.13mmol,1.0eq)溶于无水甲醇(2mL)中，随后加入碳酸钠(27mg,0.26mmol,2.0eq)，常温反应2小时。反应结束后，制备分离纯化得都黄色固体2'-氯-N-[6-(4-羟基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(17.7mg,收率27.4％)。LCMS(ESI)[M+H]+:507.1050。1H NMR(400MHz,CDCl3)δ8.80(s,1H),8.18(s,1H),7.53(s,1H),7.46(s,1H),7.14-7.07(m,2H),6.74-6.66(m,2H),3.62(s,3H),2.95-2.80(m,2H),2.69(d,J＝8.3Hz,3H),2.59(s,3H),2.07-1.80(m,2H)。4-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamido]-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)phenyl acetate (70 mg, 0.13 mmol, 1.0 eq) was dissolved in anhydrous methanol (2 mL), followed by the addition of sodium carbonate (27 mg, 0.26 mmol, 2.0 eq). The reaction was allowed to react at room temperature for 2 hours. After completion of the reaction, preparative separation and purification afforded 2'-chloro-N-[6-(4-hydroxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (17.7 mg, 27.4% yield) as a yellow solid. LCMS (ESI) [M+H] + : 507.1050. 1H NMR(400MHz, CDCl3)δ8.80(s,1H),8.18(s,1H),7.53(s,1H),7.46(s,1H),7.14-7.07(m,2H),6.74- 6.66(m,2H),3.62(s,3H),2.95-2.80(m,2H),2.69(d,J＝8.3Hz,3H),2.59(s,3H),2.07-1.80(m,2H).

实施例8：2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(化合物15)的合成
Example 8: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (Compound 15)

步骤1：8-[4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸-7-烯的合成Step 1: Synthesis of 8-[4-(trifluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]dec-7-ene

将1,4-二氧阿司匹林[4.5]癸-7-en-8-基三氟甲磺酸酯(6.0g,20.81mmol,1.0eq)和[4-(三氟甲基)苯基]硼酸(3.95g,20.81mmol,1.0eq)溶于超干1,4-二氧六环(10.0mL)中，随后加入去离子水(2.5mL)、碳酸铯(20.34g,62.4mmol,3.0eq)和二氯[1,1'-二(二苯基膦)二茂铁]钯(456.0mg,0.624mmol,0.03eq)，置换氮气，80℃反应2.5小时。反应完成后，正向柱纯化得到白色固体8-[4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸-7-烯(4.04g,收率68.35％)。LCMS(ESI)[M+H]+:285。1H NMR(400MHz,DMSO)δ7.65(m,J＝8.5Hz,4H),6.21–6.16(m,1H),3.92(s,4H),2.57(m J＝6.7,2.3Hz,2H),2.42–2.38(m,2H),1.83(t,J＝6.5Hz,2H)。1,4-Dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (6.0 g, 20.81 mmol, 1.0 eq) and [4-(trifluoromethyl)phenyl]boronic acid (3.95 g, 20.81 mmol, 1.0 eq) were dissolved in ultra-dry 1,4-dioxane (10.0 mL), followed by the addition of deionized water (2.5 mL), cesium carbonate (20.34 g, 62.4 mmol, 3.0 eq) and dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (456.0 mg, 0.624 mmol, 0.03 eq), nitrogen was replaced, and the reaction was carried out at 80°C for 2.5 hours. After completion of the reaction, forward column purification afforded 8-[4-(trifluoromethyl)phenyl]-1,4-dioxaspirino[4.5]dec-7-ene (4.04 g, 68.35% yield) as a white solid. LCMS (ESI) [M+H]+: 285. 1H NMR (400 MHz, DMSO) δ: 7.65 (m, J = 8.5 Hz, 4H), 6.21–6.16 (m, 1H), 3.92 (s, 4H), 2.57 (m, J = 6.7, 2.3 Hz, 2H), 2.42–2.38 (m, 2H), 1.83 (t, J = 6.5 Hz, 2H).

步骤2：8-[4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸烷的合成Step 2: Synthesis of 8-[4-(trifluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]decane

将8-[4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸-7-烯(850mg,2.99mmol,1.0eq)溶甲醇(15.0mL)中，加入钯/碳(31.8mg,0.299mmol,0.1eq)，随后氢气氛围下，室温反应3.0小时。反应完全后，过滤，浓缩有机相得白色固体8-[4-(三氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸烷(757mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:287。1H NMR(400MHz,DMSO)δ7.66(d,J＝8.1Hz,2H),7.48(d,J＝8.0Hz,2H),3.90(d,J＝2.3Hz,4H),2.72(m,J＝14.4,9.3,3.2Hz,1H),1.85–1.59(m,8H)。8-[4-(Trifluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]dec-7-ene (850 mg, 2.99 mmol, 1.0 eq) was dissolved in methanol (15.0 mL), and palladium/carbon (31.8 mg, 0.299 mmol, 0.1 eq) was added. The mixture was then reacted at room temperature under a hydrogen atmosphere for 3.0 hours. After the reaction was complete, the mixture was filtered and the organic phase was concentrated to obtain 8-[4-(trifluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]decane (757 mg) as a white solid, which was used directly in the next reaction. LCMS (ESI) [M+H]+: 287. 1 H NMR (400MHz, DMSO) δ7.66 (d, J = 8.1 Hz, 2H), 7.48 (d, J = 8.0 Hz, 2H), 3.90 (d, J = 2.3 Hz, 4H), 2.72 (m, J = 14.4, 9.3, 3.2 Hz, 1H), 1.85–1.59 (m, 8H).

步骤3：4-[4-(三氟甲基)苯基]环己烷-1-酮的合成Step 3: Synthesis of 4-[4-(trifluoromethyl)phenyl]cyclohexane-1-one

将8-(4-(三氟甲基)苯基)-1,4-二氧阿司匹林[4.5]癸烷(700mg,2.44mmol,1.0eq)溶于盐酸-1,4-二氧六环(4.0mL,4mol/L)中，随后室温反应16小时。反应完全后，加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯萃取，浓缩有机相得无色透明油状液体4-[4-(三氟甲基)苯基]环己烷-1-酮(603mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:243。1H NMR(400MHz,DMSO)δ7.67(d,J＝8.2Hz,2H),7.54(d,J＝8.1Hz,2H),3.18(m,J＝12.1,3.5Hz,1H),2.60(m,J＝14.2,6.0Hz,2H),2.28(m,J＝14.7,4.4,2.1Hz,2H),2.08(m,J＝12.2,5.9,2.9Hz,2H),1.92(m,J＝13.2,4.2Hz,2H)。8-(4-(Trifluoromethyl)phenyl)-1,4-dioxaspirin[4.5]decane (700 mg, 2.44 mmol, 1.0 eq) was dissolved in 1,4-dioxane hydrochloride (4.0 mL, 4 mol/L) and allowed to react at room temperature for 16 hours. After completion, saturated sodium bicarbonate solution was added to adjust the pH to neutral. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to obtain 4-[4-(trifluoromethyl)phenyl]cyclohexane-1-one (603 mg) as a colorless, transparent oil, which was used directly in the next reaction. LCMS (ESI) [M+H]+: 243. 1H NMR (400MHz, DMSO) δ7.67(d,J＝8.2Hz,2H),7.54(d,J＝8.1Hz,2H),3.18(m,J＝12.1,3.5Hz,1H),2.60(m,J＝14. 2, 6.0Hz, 2H), 2.28 (m, J = 14.7, 4.4, 2.1Hz, 2H), 2.08 (m, J = 12.2, 5.9, 2.9Hz, 2H), 1.92 (m, J = 13.2, 4.2Hz, 2H).

步骤4：6-[4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 4: Synthesis of 6-[4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-[4-(三氟甲基)苯基]环己烷-1-酮(300mg,1.24mmol,1.0eq)溶于乙腈(3mL)中，加入偶氮二异丁腈(305mg,1.86mmol,1.5eq)，0℃下加入N-溴代丁二酰亚胺(243mg,1.36mmol,1.1eq)，随后80℃反应2.0小时。反应完全后，无处理，再加入硫脲(103mg,1.36mmol,1.1eq)，80℃反应1小时后，正相柱纯化，先用乙酸乙酯洗去杂质，再用二氯化碳和甲醇体系进行淋洗，当二氯化碳和甲醇的体积比为20:1时，流出目标产物，浓缩有机相得到黄色固体6-[4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(100mg)。LCMS(ESI)[M+H]+:299。4-[4-(Trifluoromethyl)phenyl]cyclohexan-1-one (300 mg, 1.24 mmol, 1.0 eq) was dissolved in acetonitrile (3 mL), and azobisisobutyronitrile (305 mg, 1.86 mmol, 1.5 eq) was added. N-bromosuccinimide (243 mg, 1.36 mmol, 1.1 eq) was then added at 0°C, and the mixture was allowed to react at 80°C for 2.0 hours. After the reaction was complete, thiourea (103 mg, 1.36 mmol, 1.1 eq) was added without further treatment. The mixture was allowed to react at 80°C for 1 hour, and then purified on a normal phase column. Impurities were removed by washing with ethyl acetate, followed by elution with a carbon dichloride and methanol system. The desired product eluted at a carbon dichloride:methanol volume ratio of 20:1. The organic phase was concentrated to yield 6-[4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (100 mg) as a yellow solid. LCMS (ESI) [M+H] +: 299.

步骤5：2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide

将6-[4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(70.0mg,0.234mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(65.3mg,0.234mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(3mL)中，随后加入N-甲基咪唑(57.6mg,0.702mmol,3.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(98.5mg,0.35mmol,1.5eq)，室温反应2.0小时。反应完全后，制备得到白色固体2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(三氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(22.8mg收率16.89％)。LCMS(ESI)[M+H]+:559.0953。1H NMR(400MHz,DMSO)δ12.52(s,1H),8.77(s,1H),8.17(s,1H),7.68(d,J＝8.1Hz,2H),7.58(d,J＝8.1Hz,2H),7.52(s,1H),7.41(s,1H),3.62(s,3H),3.15(d,J＝9.8Hz,1H),2.96(dd,J＝16.0,5.0Hz,1H),2.84–2.70(m,3H),2.58(s,3H),2.06(t,J＝6.7Hz,2H)。6-[4-(Trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (70.0 mg, 0.234 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (65.3 mg, 0.234 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (3 mL). N-methylimidazole (57.6 mg, 0.702 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (98.5 mg, 0.35 mmol, 1.5 eq) were then added and reacted at room temperature for 2.0 hours. After the reaction was complete, a white solid 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(trifluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (22.8 mg, yield 16.89%) was obtained. LCMS (ESI) [M+H]+: 559.0953. 1H NMR (400MHz, DMSO) δ12.52(s,1H),8.77(s,1H),8.17(s,1H),7.68(d,J＝8.1Hz,2H),7.58(d,J＝8.1Hz,2H),7.52(s,1H),7.41(s ,1H),3.62(s,3H),3.15(d,J＝9.8Hz,1H),2.96(dd,J＝16.0,5.0Hz,1H),2.84–2.70(m,3H),2.58(s,3H),2.06(t,J＝6.7Hz,2H).

实施例9：2'-氯-5'-甲氧基-6-甲基-N-[6-(4-甲基噻吩-2-基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-[4,4'-联吡啶]-3-甲酰胺(化合物16)的合成
Example 9: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-[6-(4-methylthiophen-2-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-[4,4'-bipyridine]-3-carboxamide (Compound 16)

步骤1：8-(4-甲基噻吩-2-基)-1,4-二氧阿司匹林[4.5]癸-7-烯的合成Step 1: Synthesis of 8-(4-methylthiophen-2-yl)-1,4-dioxyaspirino[4.5]dec-7-ene

向1,4-二氧六环(20mL)中加入1,4-二氧阿司匹林[4.5]癸-7-en-8-基三氟甲磺酸酯(4.06g,14.09mmol,1.0eq)和(4-甲基噻吩-2-基)硼酸(2.0g,14.09mmol,1.0eq)，再加入碳酸铯(13.77g,42.26mmol,3.0eq)和水(5mL)，最后加入[1,1'-双(二苯基膦基)二茂铁]二氯化钯(514mg,0.7mmol,0.05eq)，在氮气保护下，80℃搅拌反应2小时。反应完成后，加入乙酸乙酯和水萃取反应液，收集有机层，用无水硫酸钠干燥。浓缩有机层，正相柱层析(石油醚:乙酸乙酯＝4:1)得到黄色油状物8-(4-甲基噻吩-2-基)-1,4-二氧阿司匹林[4.5]癸-7-烯(1.98g,收率59.6％)。LCMS(ESI)[M+H]+:237。1H NMR(400MHz,CDCl3)δ6.76(s,1H),6.67(d,J＝1.7Hz,1H),6.01(m,J＝4.0,1.4Hz,1H),4.00(d,J＝1.1Hz,4H),2.69–2.59(m,2H),2.47–2.39(m,2H),2.21(d,J＝1.2Hz,3H),1.90(t,J＝6.6Hz,2H)。To 1,4-dioxane (20 mL) was added 1,4-dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (4.06 g, 14.09 mmol, 1.0 eq) and (4-methylthiophen-2-yl)boronic acid (2.0 g, 14.09 mmol, 1.0 eq). Cesium carbonate (13.77 g, 42.26 mmol, 3.0 eq) and water (5 mL) were then added. Finally, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (514 mg, 0.7 mmol, 0.05 eq) was added. The mixture was stirred at 80°C under nitrogen for 2 hours. After completion of the reaction, the reaction mixture was extracted with ethyl acetate and water. The organic layer was collected and dried over anhydrous sodium sulfate. The organic layer was concentrated and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 4:1) to afford 8-(4-methylthiophen-2-yl)-1,4-dioxaspirino[4.5]dec-7-ene (1.98 g, 59.6% yield) as a yellow oil. LCMS (ESI) [M+H]+: 237. 1H NMR (400 MHz, CDCl 3 ) δ 6.76 (s, 1H), 6.67 (d, J = 1.7 Hz, 1H), 6.01 (m, J = 4.0, 1.4 Hz, 1H), 4.00 (d, J = 1.1 Hz, 4H), 2.69–2.59 (m, 2H), 2.47–2.39 (m, 2H), 2.21 (d, J = 1.2 Hz, 3H), 1.90 (t, J = 6.6 Hz, 2H).

步骤2：8-(4-甲基噻吩-2-基)-1,4-二氧杂螺[4.5]癸烷的合成Step 2: Synthesis of 8-(4-methylthiophen-2-yl)-1,4-dioxaspiro[4.5]decane

将8-(4-甲基噻吩-2-基)-1,4-二氧阿司匹林[4.5]癸-7-烯(1.00g,4.24mmol,1.0eq)溶于无水甲醇(10mL)中，再加入钯碳(100mg)，在氢气条件下，室温反应4小时。待反应完全后，将反应液过滤，浓缩滤液得到透明油状物8-(4-甲基噻吩-2-基)-1,4-二氧杂螺[4.5]癸烷粗品(920mg,产率91.3％)。8-(4-Methylthiophen-2-yl)-1,4-dioxaspiro[4.5]dec-7-ene (1.00 g, 4.24 mmol, 1.0 eq) was dissolved in anhydrous methanol (10 mL) and palladium on carbon (100 mg) was added. The mixture was reacted under hydrogen at room temperature for 4 hours. After the reaction was complete, the reaction solution was filtered and the filtrate was concentrated to obtain crude 8-(4-methylthiophen-2-yl)-1,4-dioxaspiro[4.5]decane (920 mg, 91.3% yield) as a clear oil.

步骤3：4-(4-甲基噻吩-2-基)环己烷-1-酮的合成Step 3: Synthesis of 4-(4-methylthiophen-2-yl)cyclohexane-1-one

将8-(4-甲基噻吩-2-基)-1,4-二氧杂螺[4.5]癸烷(920mg,3.87mmol,1.0eq)溶于四氢呋喃(10mL)和无水乙醇(5mL)中，再加入盐酸(5mL)，室温搅拌反应30分钟。待反应完全后，向反应液中加水(40mL)，随后在冰浴条件下，少量多次加入氢氧化钠固体，调节PH＝8～9，再加乙酸乙酯萃取，收集有机层，用无水硫酸钠干燥。浓缩有机层，正相柱层析(石油醚:乙酸乙酯＝37:3)得到黄色油状物4-(4-甲基噻吩-2-基)环己烷-1-酮(636mg,产率84.8％)。LCMS(ESI)[M+H]+:195。1H NMR(400MHz,CDCl3)δ6.73(m,J＝1.2Hz,1H),6.68(t,J＝1.1Hz,1H),3.27(m,J＝11.2,3.6Hz,1H),2.51–2.46(m,4H),2.40–2.31(m,2H),2.23(d,J＝1.1Hz,3H),2.02–1.90(m,2H)。8-(4-Methylthiophen-2-yl)-1,4-dioxaspiro[4.5]decane (920 mg, 3.87 mmol, 1.0 eq) was dissolved in tetrahydrofuran (10 mL) and anhydrous ethanol (5 mL). Hydrochloric acid (5 mL) was added and the mixture was stirred at room temperature for 30 minutes. After the reaction was complete, water (40 mL) was added to the reaction solution. Solid sodium hydroxide was then added in small portions in an ice bath to adjust the pH to 8-9. The mixture was then extracted with ethyl acetate. The organic layer was collected and dried over anhydrous sodium sulfate. The organic layer was concentrated and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 37:3) to afford 4-(4-methylthiophen-2-yl)cyclohexane-1-one (636 mg, 84.8% yield) as a yellow oil. LCMS (ESI) [M+H]+: 195. 1 H NMR (400MHz, CDCl 3 )δ6.73(m,J＝1.2Hz,1H),6.68(t,J＝1.1Hz,1H),3.27(m,J＝11.2,3.6Hz,1H),2 .51–2.46(m,4H),2.40–2.31(m,2H),2.23(d,J=1.1Hz,3H),2.02–1.90(m,2H).

步骤4：6-(4-甲基噻吩-2-基)-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 4: Synthesis of 6-(4-methylthiophen-2-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-(4-甲基噻吩-2-基)环己烷-1-酮(636mg,3.28mmol,1.0eq)和偶氮二异丁腈(808mg,4.92mmol,1.5eq)加入到乙腈(15mL)中，然后在冰浴条件下，少量多次加入N-溴代琥珀酰亚胺(875mg,4.92mmol,1.5eq)，80℃搅拌反应4.5小时。待反应完全后，冷却至室温，再加入硫脲(300mg,3.94mmol,1.2eq)，80℃搅拌反应16小时。反应完成后，浓缩反应液，正相柱层析(二氯甲烷:甲醇＝9:1)分离纯化得到棕色固体6-(4-甲基噻吩-2-基)-4,5,6,7-四氢苯并[d]噻唑-2-胺粗品(150mg,收率18.3％)。LCMS(ESI)[M+H]+:251。4-(4-Methylthiophen-2-yl)cyclohexane-1-one (636 mg, 3.28 mmol, 1.0 eq) and azobisisobutyronitrile (808 mg, 4.92 mmol, 1.5 eq) were added to acetonitrile (15 mL). N-bromosuccinimide (875 mg, 4.92 mmol, 1.5 eq) was then added in small portions under an ice bath. The mixture was stirred at 80°C for 4.5 hours. After the reaction was complete, the mixture was cooled to room temperature, and thiourea (300 mg, 3.94 mmol, 1.2 eq) was added. The mixture was stirred at 80°C for 16 hours. After completion of the reaction, the reaction mixture was concentrated and purified by normal phase column chromatography (dichloromethane:methanol = 9:1) to afford crude 6-(4-methylthiophen-2-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine as a brown solid (150 mg, 18.3% yield). LCMS (ESI) [M+H] +: 251.

步骤5：2'-氯-5'-甲氧基-6-甲基-N-[6-(4-甲基噻吩-2-基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-[6-(4-methylthiophen-2-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-[4,4'-bipyridine]-3-carboxamide

将6-(4-甲基噻吩-2-基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(100mg,0.4mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(167mg,0.6mmol,1.5eq)溶于超干的N,N-二甲基甲酰胺(3mL)中，再加入N-甲基咪唑(99mg,1.2mmol,3.0eq)，最后在冰浴条件下，加入四甲基氯代脲六氟磷酸酯(168mg,0.6mmol,1.5eq)，常温搅拌反应1小时。反应完成后，制备得到白色固体2'-氯-5'-甲氧基-6-甲基-N-[6-(4-甲基噻吩-2-基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-[4,4'-联吡啶]-3-甲酰胺(13.0mg,收率6.4％)。LCMS(ESI)[M+H]+:511.0735。1H NMR(400MHz,DMSO)δ12.50(s,1H),8.77(s,1H),8.17(s,1H),7.52(s,1H),7.41(s,1H),6.91(t,J＝1.3Hz,1H),6.78(s,1H),3.61(s,3H),3.27(d,J＝13.9Hz,1H),3.06(dd,J＝15.9,5.1Hz,1H),2.80–2.65(m,3H),2.58(s,3H),2.19(d,J＝11.5Hz,1H),2.16(d,J＝1.1Hz,3H),1.98–1.83(m,1H)。6-(4-Methylthiophen-2-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (100 mg, 0.4 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (167 mg, 0.6 mmol, 1.5 eq) were dissolved in ultra-dry N,N-dimethylformamide (3 mL). N-methylimidazole (99 mg, 1.2 mmol, 3.0 eq) was added, and finally tetramethylchlorouronium hexafluorophosphate (168 mg, 0.6 mmol, 1.5 eq) was added in an ice bath. The mixture was stirred at room temperature for 1 hour. After completion of the reaction, a white solid 2'-chloro-5'-methoxy-6-methyl-N-[6-(4-methylthiophen-2-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-[4,4'-bipyridine]-3-carboxamide (13.0 mg, yield 6.4%) was obtained. LCMS (ESI) [M+H]+: 511.0735. 1H NMR (400MHz, DMSO) δ12.50(s,1H),8.77(s,1H),8.17(s,1H),7.52(s,1H),7.41(s,1H),6.91(t,J＝1.3Hz,1H),6.78(s,1H),3.61(s,3H),3.27(d ,J＝13.9Hz,1H),3.06(dd,J＝15.9,5.1Hz,1H),2.80–2.65(m,3H),2.58( s,3H),2.19(d,J＝11.5Hz,1H),2.16(d,J＝1.1Hz,3H),1.98–1.83(m,1H).

实施例10：2'-氯-N-[6-(4-氰基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物17)的合成
Example 10: Synthesis of 2'-chloro-N-[6-(4-cyanophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 17)

步骤1：4-(1,4-二氧阿司匹林[4.5]癸-7-en-8-基)苄腈的合成Step 1: Synthesis of 4-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)benzonitrile

将1,4-二氧阿司匹林[4.5]癸-7-en-8-基三氟甲磺酸酯(3.0g,10.42mmol,1.0eq)和(4-氰基苯基)硼酸(1.68g,11.45mmol,1.1eq)溶于超干1,4-二氧六环(40.0mL)，加入去离子水(10mL)、碳酸铯(10.17g,31.22mmol,3.0eq)和二氯[1,1'-二(二苯基膦)二茂铁]钯(760mg,1.04mmol,0.1eq)，置换氮气，80℃反应4小时。反应完成后，正向纯化得到黄色固体4-(1,4-二氧阿司匹林[4.5]癸-7-en-8-基)苄腈(2.26g,收率90.0％)。LCMS(ESI)[M+H]+:242。1H NMR(400MHz,DMSO-d6)δ7.80–7.73(m,2H),7.64–7.56(m,2H),6.24(m,J＝4.1,2.4Hz,1H),3.92(s,4H),2.56(m,J＝4.5,2.3Hz,2H),2.42–2.38(m,2H),1.82(t,J＝6.5Hz,2H)。1,4-Dioxyaspirin[4.5]dec-7-en-8-yl trifluoromethanesulfonate (3.0 g, 10.42 mmol, 1.0 eq) and (4-cyanophenyl)boronic acid (1.68 g, 11.45 mmol, 1.1 eq) were dissolved in ultra-dry 1,4-dioxane (40.0 mL). Deionized water (10 mL), cesium carbonate (10.17 g, 31.22 mmol, 3.0 eq), and dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (760 mg, 1.04 mmol, 0.1 eq) were added. The atmosphere was purged with nitrogen and the reaction was carried out at 80°C for 4 hours. After completion of the reaction, forward purification was performed to obtain 4-(1,4-dioxyaspirin[4.5]dec-7-en-8-yl)benzonitrile (2.26 g, 90.0% yield) as a yellow solid. LCMS(ESI)[M+H]+:242. 1H NMR (400MHz, DMSO-d6) δ7.80–7.73(m,2H),7.64–7.56(m,2H),6.24(m,J＝4.1,2.4Hz,1 H), 3.92 (s, 4H), 2.56 (m, J = 4.5, 2.3Hz, 2H), 2.42–2.38 (m, 2H), 1.82 (t, J = 6.5Hz, 2H).

步骤2：4-(1,4-二氧阿司匹林[4.5]癸-8-基)苄腈的合成Step 2: Synthesis of 4-(1,4-dioxyaspirino[4.5]dec-8-yl)benzonitrile

将4-(1,4-二氧阿司匹林[4.5]癸-7-en-8-基)苄腈(2.2g,9.13mmol,1.0eq)溶甲醇中(60.0mL)，加入钯/碳(1.1g,0.5w.t.)，随后氢气氛围下室温反应0.5小时。反应完全后，过滤，正相分离得白色固体4-(1,4-二氧阿司匹林[4.5]癸-8-基)苄腈(1.28g,收率58.18％)。LCMS(ESI)[M+H]+:244.1H NMR(400MHz,DMSO-d6)δ7.78–7.70(m,2H),7.44(d,J＝8.1Hz,2H),3.88(d,J＝2.0Hz,4H),2.69(t,J＝10.8Hz,1H),1.76(d,J＝9.4Hz,4H),1.70–1.56(m,4H)。4-(1,4-Dioxyaspirin[4.5]dec-7-en-8-yl)benzonitrile (2.2 g, 9.13 mmol, 1.0 eq) was dissolved in methanol (60.0 mL) and palladium/carbon (1.1 g, 0.5 wt %) was added. The mixture was then reacted at room temperature under a hydrogen atmosphere for 0.5 hour. After completion of the reaction, the mixture was filtered and subjected to normal phase separation to obtain 4-(1,4-dioxyaspirin[4.5]dec-8-yl)benzonitrile (1.28 g, 58.18% yield) as a white solid. LCMS(ESI)[M+H]+: 244.1H NMR (400MHz, DMSO-d6) δ7.78–7.70(m,2H),7.44(d,J＝8.1Hz,2H),3.88(d,J＝ 2.0Hz, 4H), 2.69 (t, J = 10.8Hz, 1H), 1.76 (d, J = 9.4Hz, 4H), 1.70–1.56 (m, 4H).

步骤3：4-(4-氧代环己基)苄腈的合成Step 3: Synthesis of 4-(4-oxocyclohexyl)benzonitrile

将4-(1,4-二氧阿司匹林[4.5]癸-8-基)苄腈(500mg,2.12mmol,1.0eq)溶于盐酸二氧六环溶液中，随后室温反应16小时。反应完全后，加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯萃取，正相分离得到白色固体4-(4-氧代环己基)苄腈(840mg,收率82.35％)。LCMS(ESI)[M+H]+：200。1H NMR(400MHz,DMSO-d6)δ7.85–7.73(m,2H),7.61–7.34(m,2H),3.22–3.11(m,1H),2.58(td,J＝14.1,6.0Hz,2H),2.27(m,J＝12.9,4.7,2.5Hz,2H),2.13–2.00(m,2H),1.93(m,J＝12.9,4.1Hz,2H)。4-(1,4-Dioxyaspirino[4,5]dec-8-yl)benzonitrile (500 mg, 2.12 mmol, 1.0 eq) was dissolved in dioxane hydrochloride and allowed to react at room temperature for 16 hours. After completion, saturated sodium bicarbonate was added to adjust the pH to neutral. The mixture was extracted with ethyl acetate and subjected to normal phase separation to afford 4-(4-oxocyclohexyl)benzonitrile (840 mg, 82.35% yield) as a white solid. LCMS (ESI) [M+H]+: 200. 1H NMR (400MHz, DMSO-d6) δ7.85–7.73(m,2H),7.61–7.34(m,2H),3.22–3.11(m,1H),2.58(td,J＝14 .1,6.0Hz,2H),2.27(m,J＝12.9,4.7,2.5Hz,2H),2.13–2.00(m,2H),1.93(m,J＝12.9,4.1Hz,2H).

步骤4：4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)苄腈的合成Step 4: Synthesis of 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)benzonitrile

将4-(4-氧代环己基)苄腈(250mg,1.26mmol,1.0eq)、N-溴代丁二酰亚胺(245mg,1.38mmol,1.1eq)和偶氮二异丁腈(309mg,1.89mmol,1.5eq)加入到乙腈(10mL)中，反应12小时，反应完全后，再加入硫脲(105mg,1.26mmol,1.1eq)，80℃反应2小时，将反应液过滤，正滤液经正相柱分离后得到黄色固体粗品4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)苄腈(28mg)。LCMS(ESI)[M+H]+:256。4-(4-Oxocyclohexyl)benzonitrile (250 mg, 1.26 mmol, 1.0 eq), N-bromosuccinimide (245 mg, 1.38 mmol, 1.1 eq), and azobisisobutyronitrile (309 mg, 1.89 mmol, 1.5 eq) were added to acetonitrile (10 mL) and allowed to react for 12 hours. After the reaction was complete, thiourea (105 mg, 1.26 mmol, 1.1 eq) was added and the mixture was allowed to react at 80°C for 2 hours. The reaction solution was filtered, and the filtrate was separated by normal phase column chromatography to obtain crude 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)benzonitrile (28 mg) as a yellow solid. LCMS (ESI) [M+H]+: 256.

步骤5：2'-氯-N-[6-(4-氰基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-N-[6-(4-cyanophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)苄腈(28mg,0.11mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(30mg,0.11mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(2mL)中，加入N-甲基咪唑(27mg,0.33mmol,3.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(46mg,0.16mmol,1.5eq)，室温反应4.0小时。反应完全后，制备得白色固体2'-氯-N-[6-(4-氰基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(9.5mg,收率16.96％)。LCMS(ESI)[M+H]+:516.1180。1H NMR(400MHz,DMSO-d6)δ12.47(s,1H),8.77(s,1H),8.17(s,1H),7.85–7.75(m,2H),7.58–7.54(m,2H),7.52(s,1H),7.41(s,1H),3.61(s,3H),3.22–3.08(m,1H),2.94(dd,J＝15.9,5.0Hz,1H),2.83–2.76(m,1H),2.73(d,J＝12.2Hz,2H),2.58(s,3H),2.04(q,J＝7.2,5.6Hz,2H)。4-(2-Amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)benzonitrile (28 mg, 0.11 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (30 mg, 0.11 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (2 mL). N-Methylimidazole (27 mg, 0.33 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (46 mg, 0.16 mmol, 1.5 eq) were added and reacted at room temperature for 4.0 hours. After the reaction was complete, a white solid, 2'-chloro-N-[6-(4-cyanophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (9.5 mg, yield 16.96%), was obtained. LCMS (ESI) [M+H]+: 516.1180. 1H NMR (400MHz, DMSO-d6) δ12.47(s,1H),8.77(s,1H),8.17(s,1H),7.85–7.75(m,2H),7.58–7.54(m,2H),7.52(s,1H),7.41(s,1H),3.61(s, 3H),3.22–3.08(m,1H),2.94(dd,J＝15.9,5.0Hz,1H),2.83–2.76(m,1H),2.73(d,J＝12.2Hz,2H),2.58(s,3H),2.04(q,J＝7.2,5.6Hz,2H).

实施例11：2'-氯-5'-甲氧基-N-[6-(4-甲氧基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物19)的合成
Example 11: Synthesis of 2'-chloro-5'-methoxy-N-[6-(4-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 19)

将2'-氯-N-[6-(4-羟基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(50mg,0.098mmol,1.0eq)和碳酸钠(31.4mg,0.296mmol,3.0eq)溶于甲醇(2.0mL)中，于0℃下缓慢滴加碘甲烷(14.0mg,0.098mmol,1.0eq)，常温下反应1小时。反应完成后，加入水和乙酸乙酯萃取分液，合并有机相，制备纯化得到2'-氯-5'-甲氧基-N-[6-(4-甲氧基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-6-甲基-[4,4'-联吡啶]-3-甲酰胺(0.9mg,收率1.75％)。LCMS(ESI)[M+H]+:521.10。1H NMR(400MHz,DMSO)δ8.81(s,1H),8.18(s,1H),7.53(s,1H),7.48(s,1H),7.11(d,J＝8.2Hz,2H),6.70(d,J＝8.5Hz,2H),3.62(s,3H),2.95–2.82(m,2H),2.68(dd,J＝10.6,4.8Hz,3H),2.60(s,3H),2.03–1.86(m,2H)。2'-Chloro-N-[6-(4-hydroxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (50 mg, 0.098 mmol, 1.0 eq) and sodium carbonate (31.4 mg, 0.296 mmol, 3.0 eq) were dissolved in methanol (2.0 mL). Methyl iodide (14.0 mg, 0.098 mmol, 1.0 eq) was slowly added dropwise at 0°C and the mixture was reacted at room temperature for 1 hour. After the reaction was completed, water and ethyl acetate were added to extract the separated liquids. The organic phases were combined and purified to obtain 2'-chloro-5'-methoxy-N-[6-(4-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-6-methyl-[4,4'-bipyridine]-3-carboxamide (0.9 mg, 1.75% yield). LCMS (ESI) [M+H] + : 521.10. 1H NMR (400MHz, DMSO) δ8.81(s,1H),8.18(s,1H),7.53(s,1H),7.48(s,1H),7.11(d,J＝8.2Hz,2H),6.70(d,J＝ 8.5Hz,2H),3.62(s,3H),2.95–2.82(m,2H),2.68(dd,J＝10.6,4.8Hz,3H),2.60(s,3H),2.03–1.86(m,2H).

实施例12：2'-(二氟甲基)-5'-甲氧基-6-甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(化合物25)的合成
Example 12: Synthesis of 2'-(difluoromethyl)-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (Compound 25)

步骤1：2'-(二氟甲基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸甲酯的合成Step 1: Synthesis of methyl 2'-(difluoromethyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylate

将2-(二氟甲基)-5-甲氧基-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)吡啶(500mg,1.75mmol,1.0eq)和4-溴-6-甲基烟酸甲酯(403mg,1.75mmol,1.0eq)溶于超干1,4-二氧六环4.0mL)，随后加入水(1.0mL)、碳酸铯(1.71g,5.25mmol,3.0eq)和二氯[1,1'-二(二苯基膦)二茂铁]钯(38.4mg,0.05mmol,0.03eq)，置换氮气，80℃反应2.5小时。反应完成后，正向柱分离纯化得到无色油状液体2'-(二氟甲基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸甲酯(601mg,粗品)。LCMS(ESI)[M+H]+:309.1。1H NMR(400MHz,DMSO)δ8.88(s,1H),8.52(s,1H),7.63(s,1H),7.37(s,1H),7.15–6.80(m,1H),3.85(s,3H),3.66(s,3H),2.58(s,3H)。2-(Difluoromethyl)-5-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (500 mg, 1.75 mmol, 1.0 eq) and methyl 4-bromo-6-methylnicotinate (403 mg, 1.75 mmol, 1.0 eq) were dissolved in ultra-dry 1,4-dioxane (4.0 mL). Water (1.0 mL), cesium carbonate (1.71 g, 5.25 mmol, 3.0 eq) and dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (38.4 mg, 0.05 mmol, 0.03 eq) were then added. The atmosphere was replaced with nitrogen and the reaction was carried out at 80°C for 2.5 hours. After completion of the reaction, forward column separation and purification afforded methyl 2'-(difluoromethyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylate (601 mg, crude) as a colorless oil. LCMS (ESI) [M+H]+: 309.1. 1H NMR (400 MHz, DMSO) δ 8.88 (s, 1H), 8.52 (s, 1H), 7.63 (s, 1H), 7.37 (s, 1H), 7.15–6.80 (m, 1H), 3.85 (s, 3H), 3.66 (s, 3H), 2.58 (s, 3H).

步骤2：2'-(二氟甲基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸的合成Step 2: Synthesis of 2'-(difluoromethyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid

将2'-(二氟甲基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸甲酯(550mg,1.78mmol,1.0eq)溶于甲醇(6mL)中，加入氢氧化锂一水合物(112.5mg,2.67mmol,1.5eq)，60℃条件下反应1小时。反应结束后，将反应液冷却至室温，调节pH至中性，用乙酸乙酯萃取三次，浓缩有机相得到白色固体粗品2'-(二氟甲基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(500mg,收率95.60％)。LCMS(ESI)[M+H]+:295.08。1HNMR(400MHz,DMSO)δ13.04(s,1H),8.88(s,1H),8.50(s,1H),7.58(s,1H),7.30(s,1H),7.14–6.77(m,1H),3.85(s,3H),2.56(s,3H)。Methyl 2'-(difluoromethyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylate (550 mg, 1.78 mmol, 1.0 eq) was dissolved in methanol (6 mL) and lithium hydroxide monohydrate (112.5 mg, 2.67 mmol, 1.5 eq) was added. The mixture was reacted at 60°C for 1 hour. After completion of the reaction, the reaction solution was cooled to room temperature, the pH was adjusted to neutral, and the mixture was extracted three times with ethyl acetate. The organic phase was concentrated to afford the crude product 2'-(difluoromethyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (500 mg, 95.60% yield) as a white solid. LCMS (ESI) [M+H]+: 295.08. 1 HNMR (400MHz, DMSO) δ13.04(s,1H),8.88(s,1H),8.50(s,1H),7.58(s,1H),7.30(s,1H),7.14–6.77(m,1H),3.85(s,3H),2.56(s,3H).

步骤3：2'-(二氟甲基)-5'-甲氧基-6-甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺的合成Step 3: Synthesis of 2'-(difluoromethyl)-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide

将2'-(二氟甲基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(32mg,0.108mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(3mL)中，依次加入6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-胺(25mg,0.108mmol,1.0eq)、N-甲基咪唑(26.6mg,0.324mmol,3.0eq)和N,N,N′,N′-四甲基氯甲脒六氟磷酸盐(45.7mg,0.162mmol,1.5eq)，室温下反应2小时。反应结束后，制备得到白色固体2'-(二氟甲基)-5'-甲氧基-6-甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(32.8mg,收率59.9％)。LCMS(ESI)[M+H]+:507.1668。1H NMR(400MHz,DMSO-d6)δ12.54(s,1H),8.77(s,1H),8.46(s,1H),7.68(s,1H),7.41(s,1H),7.36–7.29(m,4H),7.22(m,J＝8.6,5.6,2.6Hz,1H),7.15–6.80(m,1H),3.69(s,3H),3.02(m,J＝9.8,5.1Hz,1H),2.91(dd,J＝16.0,5.0Hz,1H),2.80–2.65(m,3H),2.59(s,3H),2.06–1.98(m,2H)。2'-(Difluoromethyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (32 mg, 0.108 mmol, 1.0 eq) was dissolved in ultra-dry N,N-dimethylformamide (3 mL). 6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (25 mg, 0.108 mmol, 1.0 eq), N-methylimidazole (26.6 mg, 0.324 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (45.7 mg, 0.162 mmol, 1.5 eq) were added in sequence and reacted at room temperature for 2 hours. After the reaction, a white solid 2'-(difluoromethyl)-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (32.8 mg, yield 59.9%) was obtained. LCMS (ESI) [M+H] + : 507.1668. 1H NMR (400MHz, DMSO-d6) δ12.54(s,1H),8.77(s,1H),8.46(s,1H),7.68(s,1H),7.41(s,1H),7.36–7.29(m,4H),7.22(m,J＝8.6,5.6,2.6Hz,1H) ,7.15–6.80(m,1H),3.69(s,3H),3.02(m,J＝9.8,5.1Hz,1H),2.91(dd, J＝16.0,5.0Hz,1H),2.80–2.65(m,3H),2.59(s,3H),2.06–1.98(m,2H).

实施例13：2'-氯-5'-甲氧基-6-甲基-N-{6-[(四氢呋喃-3-基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(化合物28)的合成
Example 13: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[(tetrahydrofuran-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (Compound 28)

步骤1：3-(溴甲基)四氢呋喃的合成Step 1: Synthesis of 3-(bromomethyl)tetrahydrofuran

将(四氢呋喃-3-基)甲醇(1.00g,9.79mmol,1.0eq)和四溴化碳(3.85g,11.61mmol,1.2eq)超干的二氯甲烷(15mL)中，然后在冰浴条件下，加入三苯基膦(3.85g,14.68mmol,1.5eq)，常温反应16小时。反应完成后，加入二氯甲烷和水萃取反应液，收集有机层，用无水硫酸钠干燥，有机层浓缩得到黄色固体3-(溴甲基)四氢呋喃粗品。(Tetrahydrofuran-3-yl)methanol (1.00 g, 9.79 mmol, 1.0 eq) and carbon tetrabromide (3.85 g, 11.61 mmol, 1.2 eq) were added to ultra-dry dichloromethane (15 mL). Triphenylphosphine (3.85 g, 14.68 mmol, 1.5 eq) was then added under an ice bath. The mixture was allowed to react at room temperature for 16 hours. After completion of the reaction, dichloromethane and water were added to extract the reaction solution. The organic layer was collected, dried over anhydrous sodium sulfate, and concentrated to yield crude 3-(bromomethyl)tetrahydrofuran as a yellow solid.

步骤2：2'-氯-5'-甲氧基-6-甲基-N-{6-[(四氢呋喃-3-基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺的合成Step 2: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[(tetrahydrofuran-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide

将3-(溴甲基)四氢呋喃(76mg,0.47mmol,4.0eq)溶于超干的N,N-二甲基甲酰胺(4mL)中，在冰浴条件下，加入钠氢(47mg,1.16mmol,10.0eq)，再逐滴加入溶于超干的N,N-二甲基甲酰胺中的2'-氯-N-(6-羟基-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(50mg,0.12mmol,1.0eq)，100℃反应2小时。待反应完全后，过滤反应液，制备得到白色固体2'-氯-5'-甲氧基-6-甲基-N-{6-[(四氢呋喃-3-基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(5.0mg,收率8.3％)。LCMS(ESI)[M+H]+:515.1010。1H NMR(400MHz,DMSO)δ9.04(s,1H),8.11(d,J＝1.4Hz,1H),7.43(d,J＝1.4Hz,1H),7.23(s,1H),4.99(d,J＝3.8Hz,1H),4.00(s,1H),3.84–3.68(m,3H),3.65–3.59(m,1H),3.57(d,J＝1.3Hz,3H),3.51(m,J＝7.8,5.7Hz,1H),2.74(dd,J＝16.2,4.6Hz,1H),2.70–2.53(m,6H),2.40(dd,J＝16.1,6.0Hz,1H),1.93–1.71(m,3H),1.55(m,J＝9.3,5.9Hz,1H)。3-(Bromomethyl)tetrahydrofuran (76 mg, 0.47 mmol, 4.0 eq) was dissolved in ultra-dry N,N-dimethylformamide (4 mL). Sodium hydroxide (47 mg, 1.16 mmol, 10.0 eq) was added under ice-cooling conditions. 2'-Chloro-N-(6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (50 mg, 0.12 mmol, 1.0 eq) dissolved in ultra-dry N,N-dimethylformamide was added dropwise. The mixture was reacted at 100°C for 2 hours. After the reaction was complete, the reaction solution was filtered to obtain a white solid 2'-chloro-5'-methoxy-6-methyl-N-{6-[(tetrahydrofuran-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol- 2 -yl}-[4,4'-bipyridine]-3-carboxamide (5.0 mg, yield 8.3%). LCMS (ESI) [M+H]+: 515.1010. NMR (400MHz, DMSO) δ9.04(s,1H),8.11(d,J＝1.4Hz,1H),7.43(d,J＝1.4Hz,1H),7.23( s,1H),4.99(d,J＝3.8Hz,1H),4.00(s,1H),3.84–3.68(m,3H),3.65–3.59(m,1H),3.57 (d,J＝1.3Hz,3H),3.51(m,J＝7.8,5.7Hz,1H),2.74(dd,J＝16.2,4.6Hz,1H),2.70–2.5 3(m,6H),2.40(dd,J＝16.1,6.0Hz,1H),1.93–1.71(m,3H),1.55(m,J＝9.3,5.9Hz,1H).

实施例14：2'-氯-N-[6-(4-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物29)的合成
Example 14: Synthesis of 2'-chloro-N-[6-(4-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 29)

步骤1：8-(4-氟苯基)-1,4-二氧阿司匹林[4.5]癸烷的合成Step 1: Synthesis of 8-(4-fluorophenyl)-1,4-dioxyaspirino[4.5]decane

将1,4-二氧阿司匹林[4.5]癸-7-en-8-基三氟甲磺酸酯(4.0g,13.87mmol,1.0eq)和4-氟苯硼酸(2.64g,13.87mmol,1.0eq)溶于超干1,4-二氧六环(30.0mL)，随后加入去离子水(7.5mL)、碳酸铯(13.56g,41.62mmol,3.0eq)和二氯[1,1'-二(二苯基膦)二茂铁]钯(0.456g,0.624mmol,0.41eq)，置换氮气，80℃反应2.5小时。反应完成后，正向柱纯化得到白色固体8-(4-氟苯基)-1,4-二氧阿司匹林[4.5]癸烷(2.04g,收率67.46％)。LCMS(ESI)[M+H]+:235。1H NMR(400MHz,DMSO)δ7.48–7.41(m,2H),7.16–7.10(m,2H),6.00–5.96(m,1H),3.91(s,4H),2.52(s,2H),2.38–2.33(m,2H),1.80(t,J＝6.5Hz,2H)。1,4-Dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (4.0 g, 13.87 mmol, 1.0 eq) and 4-fluorophenylboronic acid (2.64 g, 13.87 mmol, 1.0 eq) were dissolved in ultra-dry 1,4-dioxane (30.0 mL). Deionized water (7.5 mL), cesium carbonate (13.56 g, 41.62 mmol, 3.0 eq), and dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (0.456 g, 0.624 mmol, 0.41 eq) were then added. The atmosphere was purged with nitrogen and the reaction was continued at 80°C for 2.5 hours. After completion of the reaction, forward column purification afforded 8-(4-fluorophenyl)-1,4-dioxyaspirino[4.5]decane (2.04 g, 67.46% yield) as a white solid. LCMS(ESI)[M+H]+:235. 1H NMR (400MHz, DMSO) δ7.48–7.41(m,2H),7.16–7.10(m,2H),6.00–5.96(m,1H),3.91(s,4H),2.52(s,2H),2.38–2.33(m,2H),1.80(t,J＝6.5Hz,2H).

步骤2：8-(4-氟苯基)-1,4-二氧阿司匹林[4.5]癸烷的合成Step 2: Synthesis of 8-(4-fluorophenyl)-1,4-dioxyaspirino[4.5]decane

将8-(4-氟苯基)-1,4-二氧阿司匹林[4.5]癸-7-烯(1.0g,4.26mmol,1.0eq)溶甲醇中(15.0mL)，随后加入钯/碳(45.4mg,0.42mmol,0.1eq)，氢气氛围下，室温反应3.0小时。反应完全后，将反应液过滤，浓缩有机相得到白色固体粗品8-(4-氟苯基)-1,4-二氧阿司匹林[4.5]癸烷(980mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:237。1H NMR(400MHz,DMSO)δ7.28–7.22(m,2H),7.12–7.05(m,2H),3.88(d,J＝2.4Hz,4H),2.58(m,J＝14.5,9.5,3.1Hz,1H),1.79–1.55(m,8H)。8-(4-Fluorophenyl)-1,4-dioxyaspirino[4.5]dec-7-ene (1.0 g, 4.26 mmol, 1.0 eq) was dissolved in methanol (15.0 mL), followed by the addition of palladium/carbon (45.4 mg, 0.42 mmol, 0.1 eq). The reaction was allowed to proceed at room temperature under a hydrogen atmosphere for 3 hours. After completion of the reaction, the reaction solution was filtered, and the organic phase was concentrated to afford crude white solid 8-(4-fluorophenyl)-1,4-dioxyaspirino[4.5]decane (980 mg), which was used directly in the next reaction. LCMS (ESI) [M+H]+: 237. 1 H NMR (400MHz, DMSO) δ7.28–7.22(m,2H),7.12–7.05(m,2H),3.88(d,J=2.4Hz,4H),2.58(m,J=14.5,9.5,3.1Hz,1H),1.79–1.55(m,8H).

步骤3；4-(4-氟苯基)环己烷-1-酮的合成Step 3: Synthesis of 4-(4-fluorophenyl)cyclohexane-1-one

将8-(4-氟苯基)-1,4-二氧阿司匹林[4.5]癸烷(500mg,2.12mmol,1.0eq)溶于四氢呋喃(4mL)和乙醇(2mL)溶液中，加入浓盐酸(2mL)，随后室温反应16小时。反应完全后，加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯萃取，浓缩有机相得无色透明油状粗品4-(4-氟苯基)环己烷-1-酮(424mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:193。1H NMR(400MHz,DMSO)δ7.38–7.29(m,2H),7.17–7.06(m,2H),3.14–3.01(m,1H),2.57(td,J＝14.1,6.0Hz,2H),2.25(m,J＝14.7,4.5,2.1Hz,2H),2.04(m,J＝12.1,5.8,2.7Hz,2H),1.86(m,J＝13.2,4.1Hz,2H)。8-(4-Fluorophenyl)-1,4-dioxaspirino[4.5]decane (500 mg, 2.12 mmol, 1.0 eq) was dissolved in tetrahydrofuran (4 mL) and ethanol (2 mL). Concentrated hydrochloric acid (2 mL) was added and the mixture was allowed to react at room temperature for 16 hours. After the reaction was complete, saturated aqueous sodium bicarbonate was added to adjust the pH to neutral. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to obtain crude 4-(4-fluorophenyl)cyclohexane-1-one (424 mg) as a colorless, transparent oil, which was used directly in the next reaction. LCMS (ESI) [M+H]+: 193. 1H NMR (400MHz, DMSO) δ7.38–7.29(m,2H),7.17–7.06(m,2H),3.14–3.01(m,1H),2.57(td,J＝14.1,6.0Hz ,2H),2.25(m,J＝14.7,4.5,2.1Hz,2H),2.04(m,J＝12.1,5.8,2.7Hz,2H),1.86(m,J＝13.2,4.1Hz,2H).

步骤4；6-(4-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 4: Synthesis of 6-(4-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-(4-氟苯基)环己烷-1-酮(200mg,1.04mmol,1.0eq)溶于乙腈(3mL)中，加入偶氮二异丁腈(256mg,1.56mmol,1.5eq)，0℃下加入N-溴代丁二酰亚胺(204mg,1.14mmol,1.1eq)，80℃反应2小时。反应完全后，无处理，再加入硫脲(87mg,1.14mmol,1.1eq)，80℃继续反应1小时，有固体析出，过滤得白色固体粗品6-(4-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(103mg)。LCMS(ESI)[M+H]+:249。1HNMR(400MHz,DMSO)δ9.22(s,2H),7.37–7.32(m,2H),7.18–7.12(m,2H),3.06–2.98(m,1H),2.61–2.55(m,4H),2.01–1.89(m,2H)。4-(4-Fluorophenyl)cyclohexane-1-one (200 mg, 1.04 mmol, 1.0 eq) was dissolved in acetonitrile (3 mL), and azobisisobutyronitrile (256 mg, 1.56 mmol, 1.5 eq) was added. N-bromosuccinimide (204 mg, 1.14 mmol, 1.1 eq) was added at 0°C, and the mixture was allowed to react at 80°C for 2 hours. After the reaction was complete, thiourea (87 mg, 1.14 mmol, 1.1 eq) was added without further treatment, and the reaction was continued at 80°C for 1 hour. A solid precipitated, which was filtered to give the crude product 6-(4-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (103 mg) as a white solid. LCMS (ESI) [M+H]+: 249. 1 HNMR (400MHz, DMSO) δ9.22(s,2H),7.37–7.32(m,2H),7.18–7.12(m,2H),3.06–2.98(m,1H),2.61–2.55(m,4H),2.01–1.89(m,2H).

步骤5；2'-氯-N-[6-(4-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-N-[6-(4-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将6-(4-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(89.2mg,0.359mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(100mg,0.359mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(3mL)中，随后加入N-甲基咪唑(88.3mg,1.16mmol,3.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(151.1mg,0.54mmol,1.5eq)，随后室温反应2.0小时。反应完全后，制备得白色固体2'-氯-N-[6-(4-氟苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(54.6mg,收率29.89％)。LCMS(ESI)[M+H]+:509.0911。1H NMR(400MHz,DMSO)δ8.80(s,1H),8.16(s,1H),7.49(s,1H),7.39–7.34(m,3H),7.17–7.10(m,2H),6.07(s,1H),3.62(s,3H),3.03(dd,J＝9.8,4.7Hz,1H),2.90(dd,J＝15.4,4.9Hz,1H),2.76–2.65(m,3H),2.57(s,3H),1.99(q,J＝8.2Hz,2H)。6-(4-Fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (89.2 mg, 0.359 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (100 mg, 0.359 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (3 mL), followed by the addition of N-methylimidazole (88.3 mg, 1.16 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (151.1 mg, 0.54 mmol, 1.5 eq), and the mixture was reacted at room temperature for 2.0 hours. After the reaction was complete, 2'-chloro-N-[6-(4-fluorophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (54.6 mg, yield 29.89%) was obtained as a white solid. LCMS (ESI) [M+H] + : 509.0911. 1H NMR (400MHz, DMSO) δ8.80(s,1H),8.16(s,1H),7.49(s,1H),7.39–7.34(m,3H),7.17–7.10(m,2H),6.07(s,1H),3.62(s ,3H),3.03(dd,J＝9.8,4.7Hz,1H),2.90(dd,J＝15.4,4.9Hz,1H),2.76–2.65(m,3H),2.57(s,3H),1.99(q,J＝8.2Hz,2H).

实施例15：5'-甲氧基-2',6'-二甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(化合物24)的合成
Example 15: Synthesis of 5'-methoxy-2',6'-dimethyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (Compound 24)

步骤1：5'-甲氧基-2',6'-二甲基-[4,4'-联吡啶]-3-羧酸的合成Step 1: Synthesis of 5'-methoxy-2',6'-dimethyl-[4,4'-bipyridine]-3-carboxylic acid

将2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(200mg,0.72mmol,1.0eq)和三甲基环三硼氧烷(3.5mol/L)(0.23mL,0.79mmol,1.1eq)溶于乙二醇二甲醚(3.5mL)中，加入碳酸钾(300mg,2.16mmol,3.0eq)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(53mg,0.072mmol,0.1eq)，在氮气保护下，封管中120℃反应8小时。反应完成后，过滤反应液，滤液反相柱层析分离纯化得到粗品5'-甲氧基-2',6'-二甲基-[4,4'-联吡啶]-3-羧酸(45mg,收率24.2％)。LCMS(ESI)[M+H]+:259。2'-Chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (200 mg, 0.72 mmol, 1.0 eq) and trimethylboroxine (3.5 mol/L) (0.23 mL, 0.79 mmol, 1.1 eq) were dissolved in ethylene glycol dimethyl ether (3.5 mL). Potassium carbonate (300 mg, 2.16 mmol, 3.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (53 mg, 0.072 mmol, 0.1 eq) were added. The mixture was reacted at 120°C in a sealed tube under nitrogen for 8 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by reverse-phase column chromatography to obtain crude 5'-methoxy-2',6'-dimethyl-[4,4'-bipyridine]-3-carboxylic acid (45 mg, 24.2% yield). LCMS (ESI) [M + H] + : 259.

步骤2：5'-甲氧基-2',6'-二甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺的合成Step 2: Synthesis of 5'-methoxy-2',6'-dimethyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide

将5'-甲氧基-2',6'-二甲基-[4,4'-联吡啶]-3-羧酸(40mg,0.16mmol,1.0eq)和6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-胺(36mg,0.16mmol,1.0eq)溶于超干的N,N-二甲基甲酰胺(2mL)中，随后加入N-甲基咪唑(76mg,0.93mmol,6.0eq)，最后在冰浴条件下加入N,N,N′,N′-四甲基氯甲脒六氟磷酸盐(65mg,0.23mmol,1.5eq)，常温反应16小时。反应结束后，制备分离纯化得到白色固体5'-甲氧基-2',6'-二甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(11.7mg,收率16.0％)。LCMS(ESI)[M+H]+:471.2464。1H NMR(400MHz,DMSO)δ12.45(s,1H),8.71(s,1H),8.18(s,1H),7.36–7.29(m,5H),7.25–7.19(m,2H),3.57(s,3H),3.09–2.96(m,1H),2.92(dd,J＝17.0,4.2Hz,1H),2.77(dd,J＝14.2,4.0Hz,1H),2.72(d,J＝5.3Hz,2H),2.57(s,3H),2.47(s,3H),2.02(m,J＝4.9Hz,2H)。5'-Methoxy-2',6'-dimethyl-[4,4'-bipyridine]-3-carboxylic acid (40 mg, 0.16 mmol, 1.0 eq) and 6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (36 mg, 0.16 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (2 mL), followed by the addition of N-methylimidazole (76 mg, 0.93 mmol, 6.0 eq). Finally, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (65 mg, 0.23 mmol, 1.5 eq) was added in an ice bath and the reaction was allowed to proceed at room temperature for 16 hours. After the reaction, preparative separation and purification were performed to obtain a white solid 5'-methoxy-2',6'-dimethyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (11.7 mg, 16.0% yield). LCMS (ESI) [M + H] + : 471.2464. 1H NMR(400MHz,DMSO)δ12.45(s,1H),8.71(s,1H),8.18(s,1H),7.36–7.29(m,5H),7.25–7.19(m,2H),3.57(s,3H),3.09–2.96(m,1H), 2.92(dd,J＝17.0,4.2Hz,1H), 2.77(dd,J＝14.2,4.0Hz,1H), 2.72(d,J＝5.3Hz,2H), 2.57(s,3H), 2.47(s,3H), 2.02(m,J＝4.9Hz,2H).

实施例16：2'-氯-N-{6-[(4,4-二氟环己基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物27)的合成
Example 16: Synthesis of 2'-chloro-N-{6-[(4,4-difluorocyclohexyl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 27)

步骤1：1,1-二氟-4-(碘甲基)环己烷的合成Step 1: Synthesis of 1,1-difluoro-4-(iodomethyl)cyclohexane

将(4,4-二氟环己基)甲醇(500mg,3.33mmol,1.0eq)溶于超干二氯甲烷(10.0mL)中，在冰浴条件下，依次加入咪唑(295mg,4.33mmol,1.3eq)、三苯基膦(1.14g,4.33mmol,1.3eq)和碘(1.1g,4.33mmol,1.3eq)，0℃下反应30分钟。反应完成后，加入饱和亚硫酸氢钠水溶液淬灭反应液，再加入乙酸乙酯萃取分液，合并有机相，浓缩得到黄色固体1,1-二氟-4-(碘甲烷)环己烷粗品(3.2g,收率100.0％)。(4,4-Difluorocyclohexyl)methanol (500 mg, 3.33 mmol, 1.0 eq) was dissolved in ultra-dry dichloromethane (10.0 mL). Imidazole (295 mg, 4.33 mmol, 1.3 eq), triphenylphosphine (1.14 g, 4.33 mmol, 1.3 eq), and iodine (1.1 g, 4.33 mmol, 1.3 eq) were added sequentially in an ice bath. The mixture was allowed to react at 0°C for 30 minutes. After completion, the reaction was quenched by the addition of saturated aqueous sodium bisulfite solution. The separated phases were extracted with ethyl acetate, and the organic phases were combined and concentrated to afford crude 1,1-difluoro-4-(iodomethane)cyclohexane as a yellow solid (3.2 g, 100.0% yield).

步骤2：6-[(4,4-二氟环己基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 2: Synthesis of 6-[(4,4-difluorocyclohexyl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将(6-羟基-4,5,6,7-四氢苯并[d]噻唑-2-基)氨基甲酸叔丁酯(300mg,1.11mmol,1.0eq)溶于超干的N,N-二甲基甲酰胺(3mL)中，在冰浴条件下加入钠氢(356mg,8.89mmol,8.0eq)，搅拌反应5分钟后，缓慢滴加溶于2mL超干的N,N-二甲基甲酰胺中的1,1-二氟-4-(碘甲烷)环己烷(2.3g,8.89mmol,8.0eq)，80℃下反应1小时。反应完成后，在冰浴条件下，滴加饱和氯化铵溶液淬灭反应液，再加入乙酸乙酯萃取分液，合并有机相，浓缩后用甲醇溶解，采用反相柱分离纯化得到透明油状物6-[(4,4-二氟环己基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(73mg,收率21.7％)。LCMS(ESI)[M+H]+:303。Dissolve tert-butyl (6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)carbamate (300 mg, 1.11 mmol, 1.0 eq) in ultra-dry N,N-dimethylformamide (3 mL). Add sodium hydroxide (356 mg, 8.89 mmol, 8.0 eq) in an ice bath. Stir and react for 5 minutes. Then, slowly add 1,1-difluoro-4-(iodomethane)cyclohexane (2.3 g, 8.89 mmol, 8.0 eq) dissolved in 2 mL of ultra-dry N,N-dimethylformamide dropwise. React at 80°C for 1 hour. After completion of the reaction, saturated ammonium chloride solution was added dropwise in an ice bath to quench the reaction mixture. Ethyl acetate was then added to extract the separated liquids. The organic phases were combined, concentrated, and then dissolved in methanol. Separation and purification using a reverse-phase column gave 6-[(4,4-difluorocyclohexyl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (73 mg, 21.7% yield) as a transparent oil. LCMS (ESI) [M+H]+: 303.

步骤3：2'-氯-N-{6-[(4,4-二氟环己基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 3: Synthesis of 2'-chloro-N-{6-[(4,4-difluorocyclohexyl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将6-[(4,4-二氟环己基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(70mg,0.23mmol 1.0eq)溶于超干N,N-二甲基甲酰胺(4mL)中，加入2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(64mg,0.23mmol,1.0eq)和N-甲基咪唑(114mg,1.39mmol,6.0eq)，随后在冰浴条件下加入N,N,N′,N′-四甲基氯甲脒六氟磷酸盐(98mg,0.35mmol,1.5eq)，室温下反应16小时。反应结束后，制备分离纯化得到白色固体2'-氯-N-(6-((4,4-二氟环己基)甲氧基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(4.0mg,收率3.1％)。LCMS(ESI)[M+H]+:563.1645。1H NMR(400MHz,DMSO)δ8.86(s,1H),8.13(s,1H),7.46(d,J＝6.2Hz,2H),7.32(s,1H),5.15(m,J＝5.1,3.7Hz,1H),3.76(s,3H),3.12–3.02(m,2H),2.83(dd,J＝16.3,4.8Hz,1H),2.56(s,3H),2.39(dd,J＝15.7,5.9Hz,2H),2.28–2.16(m,1H),1.99(d,J＝9.3Hz,2H),1.81(s,3H),1.74(d,J＝14.8Hz,4H),1.17(m,J＝13.3,7.9Hz,2H)。6-[(4,4-Difluorocyclohexyl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (70 mg, 0.23 mmol, 1.0 eq) was dissolved in ultra-dry N,N-dimethylformamide (4 mL), and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (64 mg, 0.23 mmol, 1.0 eq) and N-methylimidazole (114 mg, 1.39 mmol, 6.0 eq) were added. Then, N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (98 mg, 0.35 mmol, 1.5 eq) was added under ice-cooling conditions and the reaction was carried out at room temperature for 16 hours. After the reaction, preparative separation and purification were performed to obtain 2'-chloro-N-(6-((4,4-difluorocyclohexyl)methoxy)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (4.0 mg, 3.1% yield) as a white solid. LCMS (ESI) [M+H]+: 563.1645. 1H NMR (400MHz, DMSO) δ8.86 (s, 1H), 8.13 (s, 1H), 7.46 (d, J = 6.2Hz, 2H), 7.32 (s, 1H) ),5.15(m,J＝5.1,3.7Hz,1H),3.76(s,3H),3.12–3.02(m,2H),2.83(dd,J＝16.3,4 .8Hz,1H),2.56(s,3H),2.39(dd,J＝15.7,5.9Hz,2H),2.28–2.16(m,1H),1.99(d ,J＝9.3Hz,2H),1.81(s,3H),1.74(d,J＝14.8Hz,4H),1.17(m,J＝13.3,7.9Hz,2H).

实施例17：2'-氯-N-{6-[4-(二氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物30)的合成
Example 17: Synthesis of 2'-chloro-N-{6-[4-(difluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 30)

步骤1：8-[4-(二氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸-7-烯的合成Step 1: Synthesis of 8-[4-(difluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]dec-7-ene

将1,4-二氧阿司匹林[4.5]癸-7-en-8-基三氟甲磺酸酯(3.0g,10.4mmol,1.0eq)和4-(二氟甲基)苯基硼酸(2.64g,13.87mmol,1.0eq)溶于超干1,4-二氧六环(16.0mL)，加入去离子水(4.0mL)、碳酸铯(10.17g,31.2mmol,3.0eq)和二氯[1,1'-二(二苯基膦)二茂铁]钯(228mg,0.312mmol,0.03eq)，置换氮气三次，80℃反应2.5小时。反应完成后，正向纯化(石油醚:乙酸乙酯体积比未8:1)流出产物峰，浓缩得白色固体8-[4-(二氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸-7-烯(1.76g,收率46.21％)。LCMS(ESI)[M+H]+:267.3。1H NMR(400MHz,DMSO)δ7.53(m,J＝8.3Hz,4H),7.15–6.84(m,1H),6.14–6.10(m,1H),3.92(s,4H),2.56(m,J＝3.7,1.9Hz,2H),2.41–2.37(m,2H),1.82(t,J＝6.5Hz,2H)。1,4-Dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (3.0 g, 10.4 mmol, 1.0 eq) and 4-(difluoromethyl)phenylboronic acid (2.64 g, 13.87 mmol, 1.0 eq) were dissolved in ultra-dry 1,4-dioxane (16.0 mL), and deionized water (4.0 mL), cesium carbonate (10.17 g, 31.2 mmol, 3.0 eq) and dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (228 mg, 0.312 mmol, 0.03 eq) were added. The nitrogen atmosphere was replaced three times and the reaction was carried out at 80°C for 2.5 hours. After completion of the reaction, the product peak was purified in the forward direction (petroleum ether:ethyl acetate volume ratio 8:1) and concentrated to afford 8-[4-(difluoromethyl)phenyl]-1,4-dioxaspirino[4.5]dec-7-ene (1.76 g, 46.21% yield) as a white solid. LCMS (ESI) [M+H]+: 267.3. 1H NMR (400 MHz, DMSO) δ 7.53 (m, J = 8.3 Hz, 4H), 7.15–6.84 (m, 1H), 6.14–6.10 (m, 1H), 3.92 (s, 4H), 2.56 (m, J = 3.7, 1.9 Hz, 2H), 2.41–2.37 (m, 2H), 1.82 (t, J = 6.5 Hz, 2H).

步骤2：8-[4-(二氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸烷的合成Step 2: Synthesis of 8-[4-(difluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]decane

将8-[4-(二氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸-7-烯(1.0g,3.76mmol,1.0eq)溶甲醇(15.0mL)中，加入钯/碳(40.0mg,0.376mmol,0.1eq)，随后氢气氛围下室温反应4.0小时。反应完全后，过滤，浓缩有机相得白色固体8-[4-(二氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸烷(1.04g)，未纯化直接用于下一步反应。LCMS(ESI)[M+H]+:269.3。1H NMR(400MHz,DMSO)δ7.48(d,J＝7.9Hz,2H),7.36(d,J＝8.0Hz,2H),7.14–6.82(m,1H),3.88(d,J＝2.3Hz,4H),2.64(m,J＝14.5,9.6,3.4Hz,1H),1.78–1.58(m,8H)。8-[4-(Difluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]dec-7-ene (1.0 g, 3.76 mmol, 1.0 eq) was dissolved in methanol (15.0 mL), and palladium/carbon (40.0 mg, 0.376 mmol, 0.1 eq) was added. The mixture was then reacted at room temperature under a hydrogen atmosphere for 4 hours. After the reaction was complete, the mixture was filtered and the organic phase was concentrated to obtain 8-[4-(difluoromethyl)phenyl]-1,4-dioxyaspirino[4.5]decane (1.04 g) as a white solid, which was used in the next step without purification. LCMS (ESI) [M+H]+: 269.3. 1H NMR (400MHz, DMSO) δ7.48(d,J＝7.9Hz,2H),7.36(d,J＝8.0Hz,2H),7.14–6.82(m, 1H), 3.88 (d, J = 2.3Hz, 4H), 2.64 (m, J = 14.5, 9.6, 3.4Hz, 1H), 1.78–1.58 (m, 8H).

步骤3；4-[4-(二氟甲基)苯基]环己烷-1-酮的合成Step 3: Synthesis of 4-[4-(difluoromethyl)phenyl]cyclohexane-1-one

将8-[4-(二氟甲基)苯基]-1,4-二氧阿司匹林[4.5]癸烷(400mg,1.41mmol,1.0eq)溶于四氢呋喃/乙醇溶液(4mL/2mL)中，加入浓盐酸(1.0mL)，随后60℃反应2.0小时。反应完全后，加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯萃取，正向纯化(石油醚:乙酸乙酯＝2:1)出产物峰，浓缩有机相得黄色透明油状液体4-[4-(二氟甲基)苯基]环己烷-1-酮(263mg,收率83.22％)。LCMS(ESI)[M+H]+：225.3。1H NMR(400MHz,DMSO)δ7.53–7.42(m,4H),7.15–6.84(m,1H),3.13(m,J＝12.1,3.5Hz,1H),2.59(m,J＝14.2,6.0Hz,2H),2.27(m,J＝14.6,4.4,2.1Hz,2H),2.07(m,J＝12.2,5.9,3.0Hz,2H),1.90(m,J＝13.2,4.1Hz,2H)。8-[4-(Difluoromethyl)phenyl]-1,4-dioxaspirino[4.5]decane (400 mg, 1.41 mmol, 1.0 eq) was dissolved in a tetrahydrofuran/ethanol solution (4 mL/2 mL) and concentrated hydrochloric acid (1.0 mL) was added. The mixture was then reacted at 60°C for 2.0 hours. After completion of the reaction, the pH was adjusted to neutral by adding saturated aqueous sodium bicarbonate. The mixture was extracted with ethyl acetate and purified by forward purification (petroleum ether:ethyl acetate = 2:1) to yield the product. The organic phase was concentrated to afford 4-[4-(difluoromethyl)phenyl]cyclohexane-1-one (263 mg, 83.22% yield) as a yellow, transparent oil. LCMS (ESI) [M+H]+: 225.3. 1H NMR (400MHz, DMSO) δ7.53–7.42(m,4H),7.15–6.84(m,1H),3.13(m,J＝12.1,3.5Hz,1H),2.59(m,J＝14.2,6 .0Hz,2H),2.27(m,J＝14.6,4.4,2.1Hz,2H),2.07(m,J＝12.2,5.9,3.0Hz,2H),1.90(m,J＝13.2,4.1Hz,2H).

步骤4；6-[4-(二氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 4: Synthesis of 6-[4-(difluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-[4-(二氟甲基)苯基]环己烷-1-酮(220mg,0.98mmol,1.0eq)溶于乙腈(2mL)中，加入偶氮二异丁腈(242mg,1.47mmol,1.5eq)，0℃下加入N-溴代丁二酰亚胺(192mg,1.07mmol,1.1eq)，随后80℃反应2.0小时。反应完全后，无处理，再加入硫脲(87mg,1.14mmol,1.1eq)，80℃反应1.0小时后，产品析出，过滤得白色固体6-[4-(二氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(136mg,收率34.46％)。LCMS(ESI)[M+H]+:281。1H NMR(400MHz,DMSO)δ9.18(s,2H),7.57–7.44(m,4H),7.28–7.01(m,1H),3.08(m,J＝10.4,4.1Hz,1H),2.80(dd,J＝15.7,4.9Hz,1H),2.66–2.55(m,3H),2.04–1.91(m,2H)。4-[4-(Difluoromethyl)phenyl]cyclohexan-1-one (220 mg, 0.98 mmol, 1.0 eq) was dissolved in acetonitrile (2 mL), and azobisisobutyronitrile (242 mg, 1.47 mmol, 1.5 eq) was added. N-bromosuccinimide (192 mg, 1.07 mmol, 1.1 eq) was then added at 0°C, and the mixture was allowed to react at 80°C for 2.0 hours. After the reaction was complete, thiourea (87 mg, 1.14 mmol, 1.1 eq) was added without further treatment. After 1.0 hour at 80°C, the product precipitated and was filtered to obtain 6-[4-(difluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (136 mg, 34.46% yield) as a white solid. LCMS (ESI) [M+H]+: 281. 1H NMR (400MHz, DMSO) δ9.18 (s, 2H), 7.57–7.44 (m, 4H), 7.28–7.01 (m, 1H), 3.08 (m, J = 1 0.4, 4.1Hz, 1H), 2.80 (dd, J = 15.7, 4.9Hz, 1H), 2.66–2.55 (m, 3H), 2.04–1.91 (m, 2H).

步骤5；2'-氯-N-{6-[4-(二氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-N-{6-[4-(difluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将6-[4-(二氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(60.0mg,0.215mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(60.6mg,0.215mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(2mL)中，加入N-甲基咪唑(53mg,0.646mmol,3.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(90.8mg,0.323mmol,1.5eq)，随后室温反应2小时。反应完全后，制备得白色固体2'-氯-N-{6-[4-(二氟甲基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(17.4mg,收率14.96％)。LCMS(ESI)[M+H]+:541.1991。1H NMR(400MHz,DMSO)δ12.51(s,1H),8.77(s,1H),8.17(s,1H),7.55–7.46(m,5H),7.41(s,1H),7.17–6.85(m,1H),3.62(s,3H),3.09(d,J＝10.2Hz,1H),2.94(dd,J＝15.9,5.0Hz,1H),2.81–2.68(m,3H),2.58(s,3H),2.04(m,J＝8.0Hz,2H)。6-[4-(Difluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (60.0 mg, 0.215 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (60.6 mg, 0.215 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (2 mL), and N-methylimidazole (53 mg, 0.646 mmol, 3.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (90.8 mg, 0.323 mmol, 1.5 eq) were added, followed by reaction at room temperature for 2 hours. After the reaction was complete, 2'-chloro-N-{6-[4-(difluoromethyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (17.4 mg, yield 14.96%) was obtained as a white solid. LCMS (ESI) [M+H]+: 541.1991. 1H NMR (400MHz, DMSO) δ12.51(s,1H),8.77(s,1H),8.17(s,1H),7.55–7.46(m,5H),7.41(s,1H),7.17–6.85(m,1H),3.62 (s,3H),3.09(d,J＝10.2Hz,1H),2.94(dd,J＝15.9,5.0Hz,1H),2.81–2.68(m,3H),2.58(s,3H),2.04(m,J＝8.0Hz,2H).

实施例18：N-[6-(4-氨基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物18)的合成

Example 18: Synthesis of N-[6-(4-aminophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 18)

步骤1：[4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯基]氨基甲酸叔丁酯的合成Step 1: Synthesis of tert-butyl [4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)phenyl]carbamate

将1,4-二氧杂螺[4.5]癸-7-烯-8-基三氟甲磺酸酯(4g,13.9mmol,1.0eq)、{4-[(叔丁氧羰基)氨基]苯基}硼酸(3.62g,15.2mmol,1.1eq)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(1.02g,1.39mmol,0.1eq)和碳酸铯(9.03g,27.8mmol,2.0eq)溶于1,4-二氧六环(40mL)和水(4mL)中，氮气保护下，80℃下反应5小时。反应完全后，过滤反应液，浓缩、拌样，然后正相柱分离纯化(乙酸乙酯/石油醚，乙酸乙酯含量为30％)，得到黄色固体[4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯基]氨基甲酸叔丁酯(2.13g,收率46％)。1H NMR(400MHz,DMSO)δ9.36(s,1H),7.42(d,J＝8.4Hz,2H),7.35–7.28(m,2H),5.94(m,J＝3.9,2.0Hz,1H),3.92(s,4H),2.35(dd,J＝4.4,2.1Hz,2H),1.81(t,J＝6.4Hz,2H),1.50(d,J＝6.1Hz,11H)。1,4-Dioxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate (4 g, 13.9 mmol, 1.0 eq), {4-[(tert-butoxycarbonyl)amino]phenyl}boronic acid (3.62 g, 15.2 mmol, 1.1 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.02 g, 1.39 mmol, 0.1 eq) and cesium carbonate (9.03 g, 27.8 mmol, 2.0 eq) were dissolved in 1,4-dioxane (40 mL) and water (4 mL) and reacted at 80 ° C under nitrogen protection for 5 hours. After the reaction was complete, the reaction solution was filtered, concentrated, and stirred, then purified by normal phase column chromatography (ethyl acetate/petroleum ether, 30% ethyl acetate) to obtain tert-butyl [4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)phenyl]carbamate (2.13 g, 46% yield) as a yellow solid. 1 H NMR (400 MHz, DMSO) δ 9.36 (s, 1H), 7.42 (d, J = 8.4 Hz, 2H), 7.35–7.28 (m, 2H), 5.94 (m, J = 3.9, 2.0 Hz, 1H), 3.92 (s, 4H), 2.35 (dd, J = 4.4, 2.1 Hz, 2H), 1.81 (t, J = 6.4 Hz, 2H), 1.50 (d, J = 6.1 Hz, 11H).

步骤2：[4-(1,4-二氧杂螺[4.5]癸烷-8-基)苯基]氨基甲酸叔丁酯的合成Step 2: Synthesis of tert-butyl [4-(1,4-dioxaspiro[4.5]decan-8-yl)phenyl]carbamate

将[4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯基]氨基甲酸叔丁酯(2.13g,6.4mmol,1.0eq)、钯碳(213mg)溶于甲醇(23mL)，氢气置换三次，然后室温下反应1小时。反应完全后，混合液过滤、拌样，正相柱分离纯化(乙酸乙酯/石油醚,乙酸乙酯％＝20％)，得到白色固体[4-(1,4-二氧杂螺[4.5]癸烷-8-基)苯基]氨基甲酸叔丁酯(1.13g,3.4mmol,收率50％)。1H NMR(400MHz,DMSO)δ9.23(s,1H),7.36(d,J＝8.1Hz,2H),7.10(d,J＝8.5Hz,2H),3.89(d,J＝2.3Hz,4H),1.80–1.53(m,9H),1.48(s,9H)。Dissolve tert-butyl [4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)phenyl]carbamate (2.13 g, 6.4 mmol, 1.0 eq) and palladium on carbon (213 mg) in methanol (23 mL). The mixture was replaced with hydrogen three times and allowed to react at room temperature for 1 hour. After the reaction was complete, the mixture was filtered, stirred, and purified using a normal phase column (ethyl acetate/petroleum ether, ethyl acetate % = 20%) to obtain tert-butyl [4-(1,4-dioxaspiro[4.5]decan-8-yl)phenyl]carbamate (1.13 g, 3.4 mmol, 50% yield) as a white solid. 1 H NMR (400MHz, DMSO) δ9.23 (s, 1H), 7.36 (d, J = 8.1Hz, 2H), 7.10 (d, J = 8.5Hz, 2H), 3.89 (d, J = 2.3Hz, 4H), 1.80–1.53 (m, 9H), 1.48 (s, 9H).

步骤3：[4-(4-氧代环己基)苯基]氨基甲酸叔丁酯的合成Step 3: Synthesis of tert-butyl [4-(4-oxocyclohexyl)phenyl]carbamate

[4-(1,4-二氧杂螺[4.5]癸烷-8-基)苯基]氨基甲酸叔丁酯(1.13g,3.4mmol,1.0eq)、对甲基苯磺酸水合物(130mg,0.68mmol,0.2eq)溶于丙酮(5mL)和水(10mL)中，于60℃反应16小时。反应完全后，混合液用碳酸氢钠溶液淬灭，再用乙酸乙酯和水萃取，有机相用无水硫酸钠除去多余水分，浓缩、旋干，得到黄色固体[4-(4-氧代环己基)苯基]氨基甲酸叔丁酯(700mg,2.42mmol,收率71％)。1H NMR(400MHz,DMSO)δ9.26(s,1H),7.39(d,J＝8.2Hz,2H),7.22–7.14(m,2H),2.99(m,J＝12.0,3.4Hz,1H),2.65–2.53(m,2H),2.27(m,J＝14.6,4.4,2.1Hz,2H),2.10–1.99(m,2H),1.84(m,J＝13.2,4.1Hz,2H),1.48(s,9H)。Tert-butyl [4-(1,4-dioxaspiro[4.5]decan-8-yl)phenyl]carbamate (1.13 g, 3.4 mmol, 1.0 eq) and p-toluenesulfonic acid hydrate (130 mg, 0.68 mmol, 0.2 eq) were dissolved in acetone (5 mL) and water (10 mL) and reacted at 60°C for 16 hours. After the reaction was complete, the mixture was quenched with sodium bicarbonate solution and extracted with ethyl acetate and water. The organic phase was dehydrated with anhydrous sodium sulfate, concentrated, and dried to afford tert-butyl [4-(4-oxocyclohexyl)phenyl]carbamate (700 mg, 2.42 mmol, 71% yield) as a yellow solid. 1H NMR(400MHz,DMSO)δ9.26(s,1H),7.39(d,J＝8.2Hz,2H),7.22–7.14(m,2H),2.99(m,J＝12.0,3.4Hz,1H),2.65 –2.53(m,2H),2.27(m,J＝14.6,4.4,2.1Hz,2H),2.10–1.99(m,2H),1.84(m,J＝13.2,4.1Hz,2H),1.48(s,9H).

步骤4：[4-(3-溴-4-氧代环己基)苯基]氨基甲酸叔丁酯的合成Step 4: Synthesis of tert-butyl [4-(3-bromo-4-oxocyclohexyl)phenyl]carbamate

将[4-(4-氧代环己基)苯基]氨基甲酸叔丁酯(700mg,1.9mmol,1.0eq)溶于四氢呋喃中，加入苯基三甲基三溴化铵(715mg,1.9mmol,1.0eq)、室温反应。反应完全后，反应液用乙酸乙酯和水萃取，保留有机相，浓缩后得到[4-(3-溴-4-氧代环己基)苯基]氨基甲酸叔丁酯粗品(400mg)。LCMS(ESI)[M+H]+:268。Dissolve tert-butyl [4-(4-oxocyclohexyl)phenyl]carbamate (700 mg, 1.9 mmol, 1.0 eq) in tetrahydrofuran, add phenyltrimethylammonium tribromide (715 mg, 1.9 mmol, 1.0 eq), and react at room temperature. After completion of the reaction, extract the mixture with ethyl acetate and water. The organic phase is retained and concentrated to yield crude tert-butyl [4-(3-bromo-4-oxocyclohexyl)phenyl]carbamate (400 mg). LCMS (ESI) [M+H]+: 268.

步骤5：[4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)苯基]氨基甲酸叔丁酯的合成Step 5: Synthesis of tert-butyl [4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)phenyl]carbamate

将[4-(3-溴-4-氧代环己基)苯基]氨基甲酸叔丁酯(300mg,0.82mmol,1.0eq)溶于乙腈(5mL)，加入硫脲(63mg,0.82mmol,1.0eq)，35℃下反应3小时。反应完全后，拌样，正相柱分离纯化(二氯甲烷/甲醇，甲醇含量为10％)，得到褐色固体[4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)苯基]氨基甲酸叔丁酯(150mg,0.43mmol,收率53％)。1H NMR(400MHz,DMSO)δ9.28(s,1H),8.07(s,2H),7.40(d,J＝8.2Hz,2H),7.26–7.14(m,2H),3.12(m,J＝7.3,3.5Hz,1H),2.92(m,J＝12.3,3.6Hz,1H),2.78–2.69(m,1H),2.58–2.54(m,3H),1.97–1.88(m,1H),1.48(s,9H)。Dissolve tert-butyl [4-(3-bromo-4-oxocyclohexyl)phenyl]carbamate (300 mg, 0.82 mmol, 1.0 eq) in acetonitrile (5 mL), add thiourea (63 mg, 0.82 mmol, 1.0 eq), and react at 35°C for 3 hours. After the reaction is complete, mix the sample and purify it using a normal phase column (dichloromethane/methanol, 10% methanol content) to obtain tert-butyl [4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)phenyl]carbamate (150 mg, 0.43 mmol, 53% yield) as a brown solid. 1H NMR (400MHz, DMSO) δ9.28(s,1H),8.07(s,2H),7.40(d,J＝8.2Hz,2H),7.26–7.14(m,2H),3.12(m,J＝7.3,3.5 Hz,1H),2.92(m,J＝12.3,3.6Hz,1H),2.78–2.69(m,1H),2.58–2.54(m,3H),1.97–1.88(m,1H),1.48(s,9H).

步骤6：{4-[2-(2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺基)-4,5,6,7-四氢苯并[d]噻唑-6-基]苯基}氨基甲酸叔丁酯的合成Step 6: Synthesis of tert-butyl {4-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridyl]-3-carboxamido)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl]phenyl}carbamate

将[4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)苯基]氨基甲酸叔丁酯(60mg,0.174mmol,1.0eq)、2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(45mg,0.26mmol,2.0eq)、N,N,N,N-四甲基氯甲眯六氟磷酸盐(73mg,0.26mmol,1.5eq)和N-甲基咪唑(43mg,0.52mmol,3.0eq)溶于N,N-二甲基甲酰胺(2mL)中，室温下反应过16小时。反应完全后，浓缩反应液得到棕色油状{4-[2-(2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺基)-4,5,6,7-四氢苯并[d]噻唑-6-基]苯基}氨基甲酸叔丁酯粗品(70mg)。LCMS(ESI)[M+H]+:606.2424。Tert-butyl [4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)phenyl]carbamate (60 mg, 0.174 mmol, 1.0 eq), 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (45 mg, 0.26 mmol, 2.0 eq), N,N,N,N-tetramethylchloromethylimidazole hexafluorophosphate (73 mg, 0.26 mmol, 1.5 eq) and N-methylimidazole (43 mg, 0.52 mmol, 3.0 eq) were dissolved in N,N-dimethylformamide (2 mL) and reacted at room temperature for 16 hours. After the reaction was complete, the reaction solution was concentrated to give crude tert-butyl {4-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridyl]-3-carboxamido)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl]phenyl}carbamate (70 mg) as a brown oil. LCMS (ESI) [M+H]+: 606.2424.

步骤7：N-[6-(4-氨基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 7: Synthesis of N-[6-(4-aminophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将{4-[2-(2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺基)-4,5,6,7-四氢苯并[d]噻唑-6-基]苯基}氨基甲酸叔丁酯(65mg,0.11mmol,1.0eq)溶于三氟乙酸(0.3mL)和二氯甲烷(1mL)中，室温下反应3小时。反应完全后，制备得到白色固体N-[6-(4-氨基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(5.7mg,收率10％)。LCMS(ESI)[M+H]+:506.1417。1H NMR(400MHz,DMSO)δ12.45(s,1H),8.77(s,1H),8.17(s,1H),7.52(s,1H),7.41(s,1H),7.00–6.92(m,2H),6.55–6.47(m,2H),4.89(s,2H),3.62(s,3H),2.84(d,J＝12.0Hz,2H),2.73–2.60(m,2H),2.58(s,3H),2.53(d,J＝9.7Hz,1H),2.00–1.84(m,2H)。Tert-butyl {4-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamido)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl]phenyl}carbamate (65 mg, 0.11 mmol, 1.0 eq) was dissolved in trifluoroacetic acid (0.3 mL) and dichloromethane (1 mL) and allowed to react at room temperature for 3 hours. Upon completion of the reaction, N-[6-(4-aminophenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (5.7 mg, 10% yield) was obtained as a white solid. LCMS (ESI) [M+H]+: 506.1417. 1H NMR(400MHz,DMSO)δ12.45(s,1H),8.77(s,1H),8.17(s,1H),7.52(s,1H),7.41(s,1H),7.00–6.92(m,2H),6.55–6.47(m,2H) ,4.89(s,2H),3.62(s,3H),2.84(d,J＝12.0Hz,2H),2.73–2.60(m,2H),2.58(s,3H),2.53(d,J＝9.7Hz,1H),2.00–1.84(m,2H).

实施例19：2'-氯-5'-甲氧基-6-甲基-N-(6-吗啉基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(化合物13)的合成
Example 19: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-(6-morpholinyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (Compound 13)

步骤1：6-吗啉基-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 1: Synthesis of 6-morpholino-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-吗啉环己-1-酮(1g,5.46mmol,1.0eq)溶于乙腈(10mL)中，加入N-溴代丁二酰亚胺(972mg,5.46mmol,1.0eq)，氮气保护下，80℃反应2小时。随后降至室温，加入硫脲(415mg,1.0eq)，氮气保护下，80℃继续反应1小时。反应完成后，降至室温搅拌6小时，将反应液过滤得到黄色固体粗品6-吗啉基-4,5,6,7-四氢苯并[d]噻唑-2-胺(256mg,收率19.6％)，未经进一步纯化，直接用于下一步反应。LCMS(ESI)[M+H]+:240。4-Morpholinocyclohexan-1-one (1 g, 5.46 mmol, 1.0 eq) was dissolved in acetonitrile (10 mL), and N-bromosuccinimide (972 mg, 5.46 mmol, 1.0 eq) was added. The mixture was reacted at 80°C under nitrogen for 2 hours. The temperature was then cooled to room temperature, and thiourea (415 mg, 1.0 eq) was added. The reaction was continued at 80°C under nitrogen for 1 hour. After completion of the reaction, the mixture was cooled to room temperature and stirred for 6 hours. The reaction solution was filtered to obtain a crude yellow solid, 6-morpholino-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (256 mg, 19.6% yield). This crude product was used directly in the next step without further purification. LCMS (ESI) [M+H]+: 240.

步骤2：2'-氯-5'-甲氧基-6-甲基-N-(6-吗啉基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺的合成Step 2: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-(6-morpholino-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide

将6-吗啉基-4,5,6,7-四氢苯并[d]噻唑-2-胺(256mg,1.07mmol,1.0eq)溶于乙腈(5mL)中，加入2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(298mg,1.07mmol,1.0eq)、四甲基氯代脲六氟磷酸酯(359mg,1.21mmol,1.2eq)和N-甲基咪唑(263mg,3.21mmol,3.0eq)，氮气保护下，室温反应12小时。待反应完全后，制备分离纯化得到白色固体2'-氯-5'-甲氧基-6-甲基-N-(6-吗啉基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(8.5mg,收率1.6％)。LCMS(ESI)[M+H]+:500.15。1H NMR(400MHz,DMSO)δ12.49(s,1H),8.77(s,1H),8.18(d,J＝1.6Hz,1H),7.53(d,J＝1.6Hz,1H),7.42(s,1H),3.62(d,J＝1.6Hz,6H),2.89–2.63(m,5H),2.59(d,J＝1.6Hz,8H),2.05(s,1H),1.70(s,1H)。6-Morpholinyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (256 mg, 1.07 mmol, 1.0 eq) was dissolved in acetonitrile (5 mL), and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (298 mg, 1.07 mmol, 1.0 eq), tetramethylchlorouronium hexafluorophosphate (359 mg, 1.21 mmol, 1.2 eq) and N-methylimidazole (263 mg, 3.21 mmol, 3.0 eq) were added. The reaction was carried out at room temperature under nitrogen protection for 12 hours. After the reaction was complete, preparative separation and purification were performed to obtain a white solid 2'-chloro-5'-methoxy-6-methyl-N-(6-morpholinyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (8.5 mg, yield 1.6%). LCMS (ESI) [M+H]+: 500.15. 1H NMR (400MHz, DMSO) δ12.49(s,1H),8.77(s,1H),8.18(d,J＝1.6Hz,1H),7.53(d,J＝1.6Hz,1H),7.42 (s,1H),3.62(d,J＝1.6Hz,6H),2.89–2.63(m,5H),2.59(d,J＝1.6Hz,8H),2.05(s,1H),1.70(s,1H).

实施例20：N-[6-(6-氨基吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物20)的合成
Example 20: Synthesis of N-[6-(6-aminopyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 20)

步骤1：叔丁基[5-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)吡啶-2-基]氨基甲酸酯(的合成Step 1: Synthesis of tert-butyl[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl]carbamate

将叔丁基(5-溴吡啶-2-基)氨基甲酸酯(10g,36.6mmol,1.0eq)、联硼酸频那醇酯(13.96g,55.0mmol,1.5eq)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(2.66g,3.66mmol,0.1eq)和醋酸钾(10.76g,109.8mmol,3.0eq)溶于1,4-二氧六环(40.0mL)中，在氮气环境下，100℃反应16小时。反应结束后，收集滤液，浓缩后正相纯化得到粗产物黄色固体叔丁基[5-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)吡啶-2-基]氨基甲酸酯(5.0g,收率42.6％)。LCMS(TOF MS ES+)m/z[M+H]+:265.13。Tert-butyl (5-bromopyridin-2-yl) carbamate (10 g, 36.6 mmol, 1.0 eq), pinacol diboron (13.96 g, 55.0 mmol, 1.5 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (2.66 g, 3.66 mmol, 0.1 eq), and potassium acetate (10.76 g, 109.8 mmol, 3.0 eq) were dissolved in 1,4-dioxane (40.0 mL) and reacted at 100°C under nitrogen for 16 hours. After completion of the reaction, the filtrate was collected, concentrated, and purified by normal phase purification to obtain the crude product, tert-butyl [5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl] carbamate (5.0 g, 42.6% yield), as a yellow solid. LCMS(TOF MS ES+)m/z[M+H]+:265.13.

步骤2：[5-(1,4-二氧杂螺[4.5]癸烷-7-烯烃-8-基)吡啶-2-基]氨基甲酸酯的合成Step 2: Synthesis of [5-(1,4-dioxaspiro[4.5]dec-7-ene-8-yl)pyridin-2-yl]carbamate

将1,4-二氧杂螺[4.5]癸烷-7-烯烃-8-三氟甲磺酸酯(3.4g,10.6mmol,1.2eq)、叔丁基[5-(4,4,5,5-四甲基-1,3,2-二氧硼烷-2-基)吡啶-2-基]氨基甲酸酯(2.55g,8.85mmol,1.0eq)、碳酸铯(8.6g,26.6mmol,3.0eq)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(647mg,0.885mmol,0.1eq)溶于1,4-二氧六环(30mL)和水(3mL)的混合溶剂中，在氮气环境下，80℃反应3小时。反应结束后，将反应液过滤，滤液浓缩后正相纯化得到产物白色固体叔丁基[5-(1,4-二氧杂螺[4.5]癸烷-7-烯烃-8-基)吡啶-2-基]氨基甲酸酯(2.25g,收率63.9％)。LCMS(TOF MS ES+)m/z[M+H]+:277.11。1H NMR(400MHz,DMSO)δ9.77(s,1H),8.29(d,J＝2.4Hz,1H),7.82–7.70(m,2H),6.05–5.99(m,1H),3.91(s,4H),2.53(t,J＝4.8Hz,2H),2.39–2.33(m,2H),1.80(t,J＝6.5Hz,2H),1.47(s,9H)。1,4-Dioxaspiro[4.5]decane-7-ene-8-trifluoromethanesulfonate (3.4 g, 10.6 mmol, 1.2 eq), tert-butyl[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl]carbamate (2.55 g, 8.85 mmol, 1.0 eq), cesium carbonate (8.6 g, 26.6 mmol, 3.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (647 mg, 0.885 mmol, 0.1 eq) were dissolved in a mixed solvent of 1,4-dioxane (30 mL) and water (3 mL) and reacted at 80 ° C under nitrogen atmosphere for 3 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated and purified by normal phase chromatography to yield tert-butyl [5-(1,4-dioxaspiro[4.5]dec-7-ene-8-yl)pyridin-2-yl]carbamate (2.25 g, 63.9% yield) as a white solid. LCMS (TOF MS ES+) m/z [M+H]+: 277.11. 1H NMR (400 MHz, DMSO) δ 9.77 (s, 1H), 8.29 (d, J = 2.4 Hz, 1H), 7.82–7.70 (m, 2H), 6.05–5.99 (m, 1H), 3.91 (s, 4H), 2.53 (t, J = 4.8 Hz, 2H), 2.39–2.33 (m, 2H), 1.80 (t, J = 6.5 Hz, 2H), 1.47 (s, 9H).

步骤3：[5-(1,4-二氧杂螺[4.5]正癸烷-8-基)吡啶-2-基]癸烷的合成Step 3: Synthesis of [5-(1,4-dioxaspiro[4.5]n-decane-8-yl)pyridin-2-yl]decane

将叔丁基[5-(1,4-二氧杂螺[4.5]癸烷-7-烯烃-8-基)吡啶-2-基]氨基甲酸酯(2.2g,6.6mmol,1.0eq)溶于甲醇(20.0mL)中，加入钯碳催化剂(220mg)，氢气环境下，30℃反应16小时。反应结束后，将反应液过滤，滤液浓缩后得到白色固体叔丁基[5-(1,4-二氧杂螺[4.5]正癸烷-8-基)吡啶-2-基]癸烷(1.3g,收率59.1％)。LCMS(TOF MS ES+)m/z[M+H]+:279.12。1H NMR(400MHz,DMSO)δ9.61(s,1H),8.09(s,1H),7.69(d,J＝8.6Hz,1H),7.58(d,J＝8.7Hz,1H),3.88(d,J＝2.0Hz,4H),1.75(d,J＝10.7Hz,4H),1.62(t,J＝11.7Hz,4H),1.46(d,J＝1.9Hz,9H),1.23(s,1H)。Tert-butyl[5-(1,4-dioxaspiro[4.5]decane-7-ene-8-yl)pyridin-2-yl]carbamate (2.2 g, 6.6 mmol, 1.0 eq) was dissolved in methanol (20.0 mL) and a palladium-carbon catalyst (220 mg) was added. The mixture was reacted at 30°C under a hydrogen atmosphere for 16 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was concentrated to afford tert-butyl[5-(1,4-dioxaspiro[4.5]decane-7-ene-8-yl)pyridin-2-yl]decane (1.3 g, 59.1% yield) as a white solid. LCMS (TOF MS ES+) m/z [M+H]+: 279.12. 1H NMR (400MHz, DMSO) δ9.61(s,1H),8.09(s,1H),7.69(d,J＝8.6Hz,1H),7.58(d,J＝8.7Hz,1H),3.88(d ,J＝2.0Hz,4H),1.75(d,J＝10.7Hz,4H),1.62(t,J＝11.7Hz,4H),1.46(d,J＝1.9Hz,9H),1.23(s,1H).

步骤4：4-(6-氨基吡啶-3-基)环己酮的合成Step 4: Synthesis of 4-(6-aminopyridin-3-yl)cyclohexanone

将叔丁基[5-(1,4-二氧杂螺[4.5]正癸烷-8-基)吡啶-2-基]癸烷(1.3g,3.9mmol,1.0eq)和甲基苯磺酸一水化合物(892.7mg,4.7mmol,1.2eq)溶于丙酮(5.0mL)和水(10.0mL)的混合溶剂中，60℃下反应1小时。反应结束后，在反应液中加入饱和碳酸氢钠淬灭，用乙酸乙酯萃取三次后，浓缩后反相过柱得到黄色油状物4-(6-氨基吡啶-3-基)环己酮(600mg,收率80.6％)。LCMS(TOF MS ES+)m/z[M+H]+:191.11。1H NMR(400MHz,DMSO)δ7.82(d,J＝2.4Hz,1H),7.34(dd,J＝8.5,2.5Hz,1H),6.41(d,J＝8.4Hz,1H),5.81(s,2H),2.62–2.51(m,3H),2.23(m,J＝14.7,2.3Hz,2H),2.04–1.94(m,2H),1.80(m,J＝13.2,4.0Hz,2H)。Tert-butyl[5-(1,4-dioxaspiro[4,5-[1,5-decan-8-yl]pyridin-2-yl]decane (1.3 g, 3.9 mmol, 1.0 eq) and toluenesulfonic acid monohydrate (892.7 mg, 4.7 mmol, 1.2 eq) were dissolved in a mixture of acetone (5.0 mL) and water (10.0 mL) and reacted at 60°C for 1 hour. After completion of the reaction, the mixture was quenched with saturated sodium bicarbonate and extracted three times with ethyl acetate. The mixture was concentrated and purified by reverse phase chromatography to afford 4-(6-aminopyridin-3-yl)cyclohexanone (600 mg, 80.6% yield) as a yellow oil. LCMS (TOF MS ES+) m/z [M+H]+: 191.11. 1H NMR (400MHz, DMSO) δ7.82(d,J＝2.4Hz,1H),7.34(dd,J＝8.5,2.5Hz,1H),6.41(d,J＝8.4Hz,1H),5.81(s ,2H),2.62–2.51(m,3H),2.23(m,J＝14.7,2.3Hz,2H),2.04–1.94(m,2H),1.80(m,J＝13.2,4.0Hz,2H).

步骤5：[5-(4-氧环己基)吡啶-2-基]氨基甲酸酯的合成Step 5: Synthesis of [5-(4-oxocyclohexyl)pyridin-2-yl]carbamate

将4-(6-氨基吡啶-3-基)环己酮(400mg,2.094mmol,1.0eq)、二碳酸二叔丁酯(913mg,4.188mmol,2.0eq)和4-二甲氨基吡啶(255.5mg,2.094mmol,1.0eq)溶于四氢呋喃(15.0mL)中，室温下反应3小时。反应结束后，将反应液浓缩，采用正相柱分离纯化得到产物叔丁基[5-(4-氧环己基)吡啶-2-基]氨基甲酸酯(120mg,收率19.8％)。LCMS(TOF MS ES+)m/z[M+H]+:235.12。1H NMR(400MHz,DMSO)δ9.64(s,1H),8.18(d,J＝2.3Hz,1H),7.74–7.63(m,2H),3.04(t,J＝12.1Hz,1H),2.57(m,J＝14.2,6.0Hz,2H),2.26(d,J＝14.5Hz,2H),2.09–1.95(m,2H),1.88(m,J＝13.0,4.0Hz,2H),1.46(s,9H)。4-(6-Aminopyridin-3-yl)cyclohexanone (400 mg, 2.094 mmol, 1.0 eq), di-tert-butyl dicarbonate (913 mg, 4.188 mmol, 2.0 eq), and 4-dimethylaminopyridine (255.5 mg, 2.094 mmol, 1.0 eq) were dissolved in tetrahydrofuran (15.0 mL) and reacted at room temperature for 3 hours. After completion of the reaction, the reaction solution was concentrated and purified using a normal phase column to obtain tert-butyl [5-(4-oxocyclohexyl)pyridin-2-yl]carbamate (120 mg, 19.8% yield). LCMS (TOF MS ES+) m/z [M+H]+: 235.12. 1H NMR (400MHz, DMSO) δ9.64(s,1H),8.18(d,J＝2.3Hz,1H),7.74–7.63(m,2H),3.04(t,J＝12.1Hz,1H),2.57(m ,J＝14.2,6.0Hz,2H),2.26(d,J＝14.5Hz,2H),2.09–1.95(m,2H),1.88(m,J＝13.0,4.0Hz,2H),1.46(s,9H).

步骤6：叔丁基[5-(3-溴-4-氧环己基)吡啶-2-基]氨基甲酸酯的合成Step 6: Synthesis of tert-butyl[5-(3-bromo-4-oxocyclohexyl)pyridin-2-yl]carbamate

将叔丁基(5-(4-氧环己基)吡啶-2-基)氨基甲酸酯(80mg,0.275mmol,1.0eq)，苯基三甲基三溴化铵(103.6mg,0.275mmol,1.0eq)溶于四氢呋喃(4.0mL)中，在室温下反应过夜，反应结束后，将反应液浓缩，乙酸乙酯萃取后得到粗产物叔丁基[5-(3-溴-4-氧环己基)吡啶-2-基]氨基甲酸酯，直接进行下一步反应。LCMS(TOF MS ES+)m/z[M+H]+:369.07。Tert-butyl (5-(4-oxocyclohexyl)pyridin-2-yl)carbamate (80 mg, 0.275 mmol, 1.0 eq) and phenyltrimethylammonium tribromide (103.6 mg, 0.275 mmol, 1.0 eq) were dissolved in tetrahydrofuran (4.0 mL) and reacted overnight at room temperature. After completion of the reaction, the reaction solution was concentrated and extracted with ethyl acetate to obtain the crude product, tert-butyl [5-(3-bromo-4-oxocyclohexyl)pyridin-2-yl]carbamate, which was directly used for the next step. LCMS (TOF MS ES+) m/z [M+H]+: 369.07.

步骤7：叔丁基[5-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)吡啶-2-基]氨基甲酸酯的合成Step 7: Synthesis of tert-butyl[5-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)pyridin-2-yl]carbamate

将叔丁基[5-(3-溴-4-氧环己基)吡啶-2-基]氨基甲酸酯(101mg,0.275mmol,1.0eq)溶于乙腈(4.0mL)中，加入硫脲(21mg,0.275mmol,1.0eq)，在氮气环境下，80℃反应1小时。反应结束后，浓缩正相过柱得到黄色油状物叔丁基[5-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)吡啶-2-基]氨基甲酸酯(55.0mg,收率57.8％)。LCMS(TOF MS ES+)m/z[M+H]+:291.08。Dissolve tert-butyl [5-(3-bromo-4-oxocyclohexyl)pyridin-2-yl]carbamate (101 mg, 0.275 mmol, 1.0 eq) in acetonitrile (4.0 mL) and add thiourea (21 mg, 0.275 mmol, 1.0 eq). The mixture is reacted at 80°C under nitrogen for 1 hour. After completion of the reaction, the product is concentrated and filtered through a normal phase column to afford tert-butyl [5-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)pyridin-2-yl]carbamate (55.0 mg, 57.8% yield) as a yellow oil. LCMS (TOF MS ES+) m/z [M+H]+: 291.08.

步骤8：{5-[2-(2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺)-4,5,6,7-四氢苯并[d]噻唑-6-基]吡啶-2-基}氨基甲酸酯的合成Step 8: Synthesis of {5-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl]pyridin-2-yl}carbamate

将叔丁基[5-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)吡啶-2-基]氨基甲酸酯(50mg,0.144mmol,1.0eq)，2'-溴-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(48mg,0.173mmol,1.2eq)，N,N,N',N'-四甲基氯甲脒六氟磷酸盐(61mg,0.217mmol,1.5eq)和N-甲基咪唑(36mg,0.433mmol,3.0eq)溶于N,N-二甲基甲酰胺(2.0mL)中，氮气保护下，室温反应3小时。反应结束后，用碳酸氢钠溶液淬灭，乙酸乙酯萃取浓缩得到粗产品叔丁基{5-[2-(2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺)-4,5,6,7-四氢苯并[d]噻唑-6-基]吡啶-2-基}氨基甲酸酯(71.0mg,收率97.2％)，直接用于下一步反应。LCMS(TOF MS ES+)m/z[M+H]+:507.13。Tert-butyl [5-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)pyridin-2-yl]carbamate (50 mg, 0.144 mmol, 1.0 eq), 2'-bromo-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (48 mg, 0.173 mmol, 1.2 eq), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (61 mg, 0.217 mmol, 1.5 eq) and N-methylimidazole (36 mg, 0.433 mmol, 3.0 eq) were dissolved in N,N-dimethylformamide (2.0 mL) and reacted at room temperature under nitrogen protection for 3 hours. After the reaction, the product was quenched with sodium bicarbonate solution, extracted with ethyl acetate, and concentrated to give the crude product tert-butyl {5-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl]pyridin-2-yl}carbamate (71.0 mg, yield 97.2%), which was used directly in the next reaction. LCMS (TOF MS ES+) m/z [M+H]+: 507.13.

步骤9：N-[6-(6-氨基吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 9: Synthesis of N-[6-(6-aminopyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将叔丁基{5-[2-(2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺)-4,5,6,7-四氢苯并[d]噻唑-6-基]吡啶-2-基}氨基甲酸酯(60mg,0.09mmol,1.0eq)溶于二氯甲烷(1.0mL)中，加入三氟乙酸(0.3mL)，室温搅拌3小时。反应结束后，将反应液浓缩后，采用制备分离纯化得到白色固体N-[6-(6-氨基吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(14.4mg,收率28.9％)。LCMS(TOF MS ES+)m/z[M+H]+:507.1369。1H NMR(400MHz,DMSO)δ8.76(s,1H),8.16(s,1H),7.84(s,1H),7.51(s,1H),7.40(s,1H),7.36(d,J＝8.1Hz,1H),6.41(d,J＝8.6Hz,1H),5.75(s,2H),3.61(s,3H),2.84(d,J＝12.9Hz,2H),2.72–2.65(m,3H),2.57(s,3H),1.94(s,2H)。Tert-butyl {5-[2-(2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide)-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl]pyridin-2-yl}carbamate (60 mg, 0.09 mmol, 1.0 eq) was dissolved in dichloromethane (1.0 mL), trifluoroacetic acid (0.3 mL) was added, and the mixture was stirred at room temperature for 3 hours. After the reaction, the reaction solution was concentrated and purified by preparative separation to give N-[6-(6-aminopyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (14.4 mg, 28.9% yield) as a white solid. LCMS(TOF MS ES+)m/z[M+H]+:507.1369. 1H NMR(400MHz,DMSO)δ8.76(s,1H),8.16(s,1H),7.84(s,1H),7.51(s,1H),7.40(s,1H),7.36(d,J＝8.1Hz,1H),6.41 (d,J＝8.6Hz,1H),5.75(s,2H),3.61(s,3H),2.84(d,J＝12.9Hz,2H),2.72–2.65(m,3H),2.57(s,3H),1.94(s,2H).

实施例21：2'-氯-N-(6-羟基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物22)的合成
Example 21: Synthesis of 2'-chloro-N-(6-hydroxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 22)

步骤1：8-苯基-1,4-二恶螺[4.5]癸-8-醇的合成Step 1: Synthesis of 8-phenyl-1,4-dioxaspiro[4.5]decan-8-ol

将1,4-二氧阿司匹林[4.5]癸-8-酮(5.00g,32mmol,1.0eq)溶于超干四氢呋喃(50.0mL)中，在冰浴条件下，逐滴加入苯基溴化镁的四氢呋喃溶液(1mol/L)(35.22mL,35.22mmol,1.1eq)，常温反应1小时。反应完成后，在冰浴条件下，加入饱和氯化铵水溶液淬灭反应液，加入乙酸乙酯萃取分液，浓缩有机相，正相柱分离纯化得到白色固体8-苯基-1,4-二恶螺[4.5]癸-8-醇(1.9g,收率25.4％)。1H NMR(400MHz,DMSO)δ7.49–7.42(m,2H),7.30(dd,J＝8.4,7.0Hz,2H),7.23–7.16(m,1H),4.89(s,1H),3.88(d,J＝2.1Hz,4H),2.01–1.85(m,4H),1.64(dd,J＝9.6,2.5Hz,2H),1.58–1.46(m,2H)。1,4-Dioxaspirino[4.5]decan-8-one (5.00 g, 32 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (50.0 mL). Phenylmagnesium bromide (1 mol/L) in tetrahydrofuran (35.22 mL, 35.22 mmol, 1.1 eq) was added dropwise in an ice bath. The reaction was allowed to react at room temperature for 1 hour. After completion, the reaction was quenched with saturated aqueous ammonium chloride in an ice bath. The mixture was extracted with ethyl acetate, and the organic phase was concentrated. Purification by normal phase column chromatography afforded 8-phenyl-1,4-dioxaspiro[4.5]decan-8-ol (1.9 g, 25.4% yield) as a white solid. 1H NMR (400MHz, DMSO) δ7.49–7.42(m,2H),7.30(dd,J＝8.4,7.0Hz,2H),7.23–7.16(m,1H),4.89(s ,1H),3.88(d,J＝2.1Hz,4H),2.01–1.85(m,4H),1.64(dd,J＝9.6,2.5Hz,2H),1.58–1.46(m,2H).

步骤2：4-羟基-4-苯基环己烷-1-酮的合成Step 2: Synthesis of 4-hydroxy-4-phenylcyclohexane-1-one

将8-苯基-1,4-二恶螺[4.5]癸-8-醇(500mg,2.14mmol,1.0eq)溶于四氢呋喃(6mL)和乙醇(3mL)中，在冰浴条件下逐滴加入浓盐酸(3mL)，在0℃下反应30分钟后。反应完成后，在冰浴条件下，加入饱和碳酸氢钠水溶液调节PH＝7～8，加入乙酸乙酯萃取分液，浓缩有机相，得到白色固体粗品4-羟基-4-苯基环己烷-1-酮(480mg,收率100.00％)。8-Phenyl-1,4-dioxaspiro[4.5]decan-8-ol (500 mg, 2.14 mmol, 1.0 eq) was dissolved in tetrahydrofuran (6 mL) and ethanol (3 mL). Concentrated hydrochloric acid (3 mL) was added dropwise in an ice bath. The mixture was reacted at 0°C for 30 minutes. After completion of the reaction, saturated aqueous sodium bicarbonate solution was added in an ice bath to adjust the pH to 7-8. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to obtain a crude white solid of 4-hydroxy-4-phenylcyclohexane-1-one (480 mg, 100.00% yield).

步骤3：2-溴-4-羟基-4-苯基环己烷-1-酮的合成Step 3: Synthesis of 2-bromo-4-hydroxy-4-phenylcyclohexane-1-one

将4-甲氧基-4-苯基环己烷-1-酮(200mg,1.05mmol,1.0eq)溶于超干四氢呋喃(3mL)中，在冰浴条件下逐滴加入溶于超干四氢呋喃(3mL)中的苯基三甲基三溴化铵(396mg,1.05mmol,1.0eq)，在室温下反应3小时。反应完成后，向反应液中加入水和乙酸乙酯萃取分液，浓缩有机相，得到棕色油状物粗品2-溴-4-羟基-4-苯基环己烷-1-酮粗品(290mg,收率100.0％)。4-Methoxy-4-phenylcyclohexane-1-one (200 mg, 1.05 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (3 mL). Phenyltrimethylammonium tribromide (396 mg, 1.05 mmol, 1.0 eq) dissolved in ultra-dry tetrahydrofuran (3 mL) was added dropwise in an ice bath. The mixture was allowed to react at room temperature for 3 hours. After completion of the reaction, water and ethyl acetate were added to the reaction mixture, and the organic phase was concentrated to afford crude 2-bromo-4-hydroxy-4-phenylcyclohexane-1-one (290 mg, 100.0% yield) as a brown oil.

步骤4：2-氨基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-6-醇的合成Step 4: Synthesis of 2-amino-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-6-ol

将2-溴-4-羟基-4-苯基环己烷-1-酮溶于乙腈(4mL)中，再加入硫脲(120mg,1.58mmol,1.5eq)，在室温下反应16小时。反应完成后，浓缩反应液，得到棕色油状物2-氨基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-6-醇粗品(240mg,收率92.7％)。LCMS(ESI)[M+H]+:247。2-Bromo-4-hydroxy-4-phenylcyclohexan-1-one was dissolved in acetonitrile (4 mL), and thiourea (120 mg, 1.58 mmol, 1.5 eq) was added. The mixture was allowed to react at room temperature for 16 hours. After completion, the reaction solution was concentrated to afford crude 2-amino-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-6-ol (240 mg, 92.7% yield) as a brown oil. LCMS (ESI) [M+H]+: 247.

步骤5：(R)-2'-氯-N-(6-羟基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of (R)-2'-chloro-N-(6-hydroxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(117mg,0.422mmol 1.0eq)溶于超干N,N-二甲基甲酰胺(3mL)中，再加入2-氨基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-6-醇(108mg,0.422mmol,1.0eq)和N-甲基咪唑(208mg,2.53mmol,6.0eq)，随后在冰浴条件下加入N,N,N′,N′-四甲基氯甲脒六氟磷酸盐(178mg,0.633mmol,1.5eq)，室温下反应2小时。反应结束后，制备分离纯化得到消旋体化合物2'-氯-N-(6-羟基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(37.7mg,收率17.7％)。将消旋体再经手性分离柱进一步纯化，得到(R)-2'-氯-N-(6-羟基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(5.6mg)和(S)-2'-氯-N-(6-羟基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(5.6mg)。LCMS(ESI)[M+H]+:507.1248。1HNMR(400MHz,DMSO)δ8.82(s,1H),8.19(s,1H),7.57–7.50(m,3H),7.47(s,1H),7.33(dd,J＝8.3,6.8Hz,2H),7.27–7.20(m,1H),3.63(s,3H),3.09(d,J＝16.5Hz,1H),2.79(d,J＝16.5Hz,2H),2.60(s,3H),2.54(s,1H),2.21(m,J＝11.9,5.7Hz,1H),1.92(d,J＝12.6Hz,1H)。2'-Chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (117 mg, 0.422 mmol, 1.0 eq) was dissolved in ultra-dry N,N-dimethylformamide (3 mL), and 2-amino-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-6-ol (108 mg, 0.422 mmol, 1.0 eq) and N-methylimidazole (208 mg, 2.53 mmol, 6.0 eq) were added. N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (178 mg, 0.633 mmol, 1.5 eq) was then added under ice-cooling conditions and the reaction was carried out at room temperature for 2 hours. After the reaction, the racemic compound 2'-chloro-N-(6-hydroxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (37.7 mg, yield 17.7%) was obtained by preparative separation and purification. The racemate was further purified by chiral separation column to give (R)-2'-chloro-N-(6-hydroxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (5.6 mg) and (S)-2'-chloro-N-(6-hydroxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (5.6 mg). LCMS (ESI) [M+H]+: 507.1248. 1 HNMR(400MHz,DMSO)δ8.82(s,1H),8.19(s,1H),7.57–7.50(m,3H),7.47(s,1H),7.33(dd,J＝8.3,6.8Hz,2H),7.27–7.20(m,1H),3.63( s,3H),3.09(d,J＝16.5Hz,1H),2.79(d,J＝16.5Hz,2H),2.60(s,3H),2.54(s,1H),2.21(m,J＝11.9,5.7Hz,1H),1.92(d,J＝12.6Hz,1H).

实施例22：2'-氯-5'-甲氧基-N-(6-甲氧基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物23)的合成
Example 22: Synthesis of 2'-chloro-5'-methoxy-N-(6-methoxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 23)

步骤1：8-苯基-1,4-二恶螺[4.5]癸-8-醇的合成Step 1: Synthesis of 8-phenyl-1,4-dioxaspiro[4.5]decan-8-ol

将1,4-二氧阿司匹林[4.5]癸-8-酮(5.00g,32mmol,1.0eq)溶于超干四氢呋喃(50.0mL)中，在冰浴条件下，逐滴加入苯基溴化镁的四氢呋喃溶液(1mol/L)(35.22mL,35.22mmol,1.1eq)，常温反应1小时。反应完成后，在冰浴条件下，加入饱和氯化铵水溶液淬灭反应液，加入乙酸乙酯萃取分液，浓缩有机相，正相柱分离纯化得到白色固体8-苯基-1,4-二恶螺[4.5]癸-8-醇(1.9g,收率25.4％)。1H NMR(400MHz,DMSO)δ7.49–7.42(m,2H),7.30(dd,J＝8.4,7.0Hz,2H),7.23–7.16(m,1H),4.89(s,1H),3.88(d,J＝2.1Hz,4H),2.01–1.85(m,4H),1.64(dd,J＝9.6,2.5Hz,2H),1.58–1.46(m,2H)。1,4-Dioxaspirino[4.5]decan-8-one (5.00 g, 32 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (50.0 mL). Phenylmagnesium bromide (1 mol/L) in tetrahydrofuran (35.22 mL, 35.22 mmol, 1.1 eq) was added dropwise in an ice bath. The reaction was allowed to react at room temperature for 1 hour. After completion, the reaction was quenched with saturated aqueous ammonium chloride in an ice bath. The mixture was extracted with ethyl acetate, and the organic phase was concentrated. Purification by normal phase column chromatography afforded 8-phenyl-1,4-dioxaspiro[4.5]decan-8-ol (1.9 g, 25.4% yield) as a white solid. 1H NMR (400MHz, DMSO) δ7.49–7.42(m,2H),7.30(dd,J＝8.4,7.0Hz,2H),7.23–7.16(m,1H),4.89(s ,1H),3.88(d,J＝2.1Hz,4H),2.01–1.85(m,4H),1.64(dd,J＝9.6,2.5Hz,2H),1.58–1.46(m,2H).

步骤2：8-甲氧基-8-苯基-1,4-二氧杂螺[4.5]癸烷的合成Step 2: Synthesis of 8-methoxy-8-phenyl-1,4-dioxaspiro[4.5]decane

将8-苯基-1,4-二恶螺[4.5]癸-8-醇(800mg,3.42mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(6mL)中，在冰浴条件下加入钠氢(410mg,10.26mmol,3.0eq)，在0℃下反应10分钟后，加入碘甲烷(728mg,5.13mmol,1.5eq)，在50℃下反应16小时。反应完成后，在冰浴条件下，加入饱和氯化铵水溶液淬灭反应液，加入乙酸乙酯萃取分液，浓缩有机相，得到淡黄色固体8-甲氧基-8-苯基-1,4-二氧杂螺[4.5]癸烷(458mg,收率54.0％)。1H NMR(400MHz,DMSO)δ7.41–7.32(m,4H),7.26(m,J＝8.5,5.9,2.0Hz,1H),3.88(s,4H),2.87(s,3H),2.02–1.94(m,2H),1.93–1.75(m,4H),1.63–1.50(m,2H)。8-Phenyl-1,4-dioxaspiro[4.5]decan-8-ol (800 mg, 3.42 mmol, 1.0 eq) was dissolved in ultra-dry N,N-dimethylformamide (6 mL). Sodium hydroxide (410 mg, 10.26 mmol, 3.0 eq) was added under ice-cooling conditions. The mixture was reacted at 0°C for 10 minutes, followed by the addition of iodomethane (728 mg, 5.13 mmol, 1.5 eq). The mixture was allowed to react at 50°C for 16 hours. After completion of the reaction, saturated aqueous ammonium chloride was added to quench the reaction mixture under ice-cooling conditions. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to obtain 8-methoxy-8-phenyl-1,4-dioxaspiro[4.5]decane (458 mg, 54.0% yield) as a pale yellow solid. 1H NMR (400MHz, DMSO) δ7.41–7.32(m,4H),7.26(m,J＝8.5,5.9,2.0Hz,1H),3.88( s,4H),2.87(s,3H),2.02–1.94(m,2H),1.93–1.75(m,4H),1.63–1.50(m,2H).

步骤3：4-甲氧基-4-苯基环己烷-1-酮的合成Step 3: Synthesis of 4-methoxy-4-phenylcyclohexane-1-one

将8-甲氧基-8-苯基-1,4-二氧杂螺[4.5]癸烷(700mg,2.82mmol,1.0eq)溶于四氢呋喃(6mL)和乙醇(3mL)中，在冰浴条件下逐滴加入浓盐酸(3mL)，室温下反应16小时后。反应完成后，在冰浴条件下，加入饱和碳酸氢钠水溶液调节PH＝7～8，加入乙酸乙酯萃取分液，浓缩有机相，得到淡黄色油状物4-甲氧基-4-苯基环己烷-1-酮(536mg,收率93.1％)。8-Methoxy-8-phenyl-1,4-dioxaspiro[4.5]decane (700 mg, 2.82 mmol, 1.0 eq) was dissolved in tetrahydrofuran (6 mL) and ethanol (3 mL). Concentrated hydrochloric acid (3 mL) was added dropwise in an ice bath. The mixture was allowed to react at room temperature for 16 hours. After completion of the reaction, saturated aqueous sodium bicarbonate was added to adjust the pH to 7-8 in an ice bath. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to afford 4-methoxy-4-phenylcyclohexane-1-one (536 mg, 93.1% yield) as a pale yellow oil.

步骤4：2-溴-4-甲氧基-4-苯基环己烷-1-酮的合成Step 4: Synthesis of 2-bromo-4-methoxy-4-phenylcyclohexane-1-one

将4-甲氧基-4-苯基环己烷-1-酮(180mg,0.88mmol,1.0eq)溶于超干四氢呋喃(3mL)中，在冰浴条件下，逐滴加入溶于超干四氢呋喃(3mL)的苯基三甲基三溴化铵(332mg,0.88mmol,3.0eq)，在室温下反应3小时。反应完成后，向反应液中加入水和乙酸乙酯萃取分液，浓缩有机相，得到淡黄色固体2-溴-4-甲氧基-4-苯基环己烷-1-酮粗品(270mg,收率100.0％)。4-Methoxy-4-phenylcyclohexane-1-one (180 mg, 0.88 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (3 mL). Phenyltrimethylammonium tribromide (332 mg, 0.88 mmol, 3.0 eq) dissolved in ultra-dry tetrahydrofuran (3 mL) was added dropwise in an ice bath. The mixture was allowed to react at room temperature for 3 hours. After completion of the reaction, water and ethyl acetate were added to the reaction mixture, and the organic phase was concentrated to afford crude 2-bromo-4-methoxy-4-phenylcyclohexane-1-one (270 mg, 100.0% yield) as a pale yellow solid.

步骤5：6-甲氧基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 5: Synthesis of 6-methoxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将8-苯基-1,4-二恶螺[4.5]癸-8-醇粗品溶于乙腈(5mL)中，再加入硫脲(134mg,1.76mmol,2.0eq)，在室温下反应16小时。反应完成后，浓缩反应液，得到棕色固体6-甲氧基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-胺粗品(240mg,收率100.0％)。LCMS(ESI)[M+H]+:261。Crude 8-phenyl-1,4-dioxaspiro[4.5]decan-8-ol was dissolved in acetonitrile (5 mL) and thiourea (134 mg, 1.76 mmol, 2.0 eq) was added. The mixture was allowed to react at room temperature for 16 hours. After completion, the reaction solution was concentrated to afford crude 6-methoxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine as a brown solid (240 mg, 100.0% yield). LCMS (ESI) [M+H]+: 261.

步骤6：(R)-2'-氯-5'-甲氧基-N-(6-甲氧基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of (R)-2'-chloro-5'-methoxy-N-(6-methoxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-6-methyl-[4,4'-bipyridine]-3-carboxamide

将2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(160mg,0.577mmol,1.5eq)溶于乙腈(3mL)中，再加入6-甲氧基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-胺(100mg,0.384mmol,1.0eq)和N-甲基咪唑(189mg,2.308mmol,6.0eq)，随后在冰浴条件下加入N,N,N′,N′-四甲基氯甲脒六氟磷酸盐(162mg,0.577mmol,1.5eq)，室温下反应1小时。反应结束后，制备分离纯化得到消旋体化合物2'-氯-5'-甲氧基-N-(6-甲氧基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-6-甲基-[4,4'-联吡啶]-3-甲酰胺(45.0mg)。消旋体化合物再经手性柱分离纯化得到(R)-2'-氯-5'-甲氧基-N-(6-甲氧基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-6-甲基-[4,4'-联吡啶]-3-甲酰胺(10.5mg)和(S)-2'-氯-5'-甲氧基-N-(6-甲氧基-6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-6-甲基-[4,4'-联吡啶]-3-甲酰胺(10.5mg)。LCMS(ESI)[M+H]+:521.1406。1H NMR(400MHz,DMSO)δ12.47(s,1H),8.76(s,1H),8.17(s,1H),7.51(s,1H),7.46–7.33(m,5H),7.29(t,J＝7.0Hz,1H),3.62(s,3H),3.14(d,J＝16.6Hz,1H),3.00(d,J＝16.5Hz,1H),2.92(s,3H),2.68(dt,J＝15.1,7.0Hz,1H),2.57(s,3H),2.41(dt,J＝16.3,5.2Hz,1H),2.35–2.17(m,2H)。2'-Chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (160 mg, 0.577 mmol, 1.5 eq) was dissolved in acetonitrile (3 mL), and 6-methoxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (100 mg, 0.384 mmol, 1.0 eq) and N-methylimidazole (189 mg, 2.308 mmol, 6.0 eq) were added. Then, N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (162 mg, 0.577 mmol, 1.5 eq) was added under ice-cooling conditions and the reaction was carried out at room temperature for 1 hour. After the reaction, the racemic compound 2'-chloro-5'-methoxy-N-(6-methoxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-6-methyl-[4,4'-bipyridine]-3-carboxamide (45.0 mg) was obtained by preparative separation and purification. The racemic compound was then separated and purified by chiral column to give (R)-2'-chloro-5'-methoxy-N-(6-methoxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-6-methyl-[4,4'-bipyridine]-3-carboxamide (10.5 mg) and (S)-2'-chloro-5'-methoxy-N-(6-methoxy-6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-6-methyl-[4,4'-bipyridine]-3-carboxamide (10.5 mg). LCMS (ESI) [M+H]+: 521.1406. 1H NMR (400MHz, DMSO) δ12.47(s,1H),8.76(s,1H),8.17(s,1H),7.51(s,1H),7.46–7.33(m,5H),7.29(t,J＝7.0Hz,1H),3.62(s,3H),3.14(d,J＝1 6.6Hz,1H),3.00(d,J＝16.5Hz,1H),2.92(s,3H),2.68(dt,J＝15.1,7.0Hz,1H),2.57(s,3H),2.41(dt,J＝16.3,5.2Hz,1H),2.35–2.17(m,2H).

实施例23：2'-氨基-5'-甲氧基-6-甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(化合物26)的合成
Example 23: Synthesis of 2'-amino-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (Compound 26)

步骤1：2'-氨基-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸的合成Step 1: Synthesis of 2'-amino-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid

将2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(255mg,0.92mmol,1.0eq)溶于超干四氢呋喃(5.0mL)中，在冰浴条件下，逐滴加入双(三甲基硅基)氨基锂的四氢呋喃溶液(1mol/L)(4.59mL,4.59mmol,5.0eq)，再加入三(二亚苄基丙酮)二钯(84mg,0.09mmol,0.1eq)和2-(二环己基膦基)联苯(64mg,0.18mmol,0.2eq)，在氮气保护下，65℃反应1.5小时。反应完成后，加入饱和氯化铵水溶液淬灭反应液，浓缩体系，加入N,N-二甲基甲酰胺(4mL)，过滤，滤液反相柱分离纯化得到棕色固体2'-氨基-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(20mg,收率8.4％)。2'-Chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (255 mg, 0.92 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (5.0 mL). Under ice bath conditions, a tetrahydrofuran solution of lithium bis(trimethylsilyl)amide (1 mol/L) (4.59 mL, 4.59 mmol, 5.0 eq) was added dropwise. Tris(dibenzylideneacetone)dipalladium (84 mg, 0.09 mmol, 0.1 eq) and 2-(dicyclohexylphosphino)biphenyl (64 mg, 0.18 mmol, 0.2 eq) were then added. The reaction was carried out at 65°C under nitrogen protection for 1.5 hours. After the reaction was completed, saturated aqueous ammonium chloride solution was added to quench the reaction solution, the system was concentrated, N,N-dimethylformamide (4 mL) was added, and the mixture was filtered. The filtrate was separated and purified by reverse phase column to obtain a brown solid 2'-amino-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (20 mg, yield 8.4%).

步骤2：2'-氨基-5'-甲氧基-6-甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺的合成Step 2: Synthesis of 2'-amino-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide

将2'-氨基-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(20mg,0.077mmol 1.0eq)溶于超干N,N-二甲基甲酰胺(1mL)中，加入乙腈(2mL)、6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-胺(18mg,0.077mmol,1.0eq)和N-甲基咪唑(38mg,0.463mmol,6.0eq)，随后在冰浴条件下，加入N,N,N′,N′-四甲基氯甲脒六氟磷酸盐(32mg,0.116mmol,1.5eq)，室温下反应16小时。反应结束后，制备分离纯化得到白色固体2'-氨基-5'-甲氧基-6-甲基-N-(6-苯基-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(6.6mg,收率18.1％)。LCMS(ESI)[M+H]+:472.1797。1H NMR(400MHz,DMSO)δ8.68(s,1H),7.63(s,1H),7.36–7.29(m,4H),7.27(s,1H),7.24–7.18(m,1H),6.43(s,1H),5.64(s,2H),3.44(s,3H),3.02(m,J＝6.7Hz,1H),2.91(dd,J＝16.1,5.3Hz,1H),2.80–2.65(m,3H),2.56(s,3H),2.02(m,J＝8.4Hz,2H)。2'-Amino-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (20 mg, 0.077 mmol, 1.0 eq) was dissolved in ultra-dry N,N-dimethylformamide (1 mL). Acetonitrile (2 mL), 6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (18 mg, 0.077 mmol, 1.0 eq) and N-methylimidazole (38 mg, 0.463 mmol, 6.0 eq) were added. N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (32 mg, 0.116 mmol, 1.5 eq) was then added in an ice bath. The reaction was allowed to proceed at room temperature for 16 hours. After the reaction, preparative separation and purification were performed to obtain a white solid 2'-amino-5'-methoxy-6-methyl-N-(6-phenyl-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (6.6 mg, yield 18.1%). LCMS (ESI) [M+H]+: 472.1797. 1H NMR(400MHz,DMSO)δ8.68(s,1H),7.63(s,1H),7.36–7.29(m,4H),7.27(s,1H),7.24–7.18(m,1H),6.43(s,1H),5.64(s,2H ),3.44(s,3H),3.02(m,J＝6.7Hz,1H),2.91(dd,J＝16.1,5.3Hz,1H),2.80–2.65(m,3H),2.56(s,3H),2.02(m,J＝8.4Hz,2H).

实施例24：2'-氯-N-[6-(4-氰基苯基)-6-羟基-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物32)的合成
Example 24: Synthesis of 2'-chloro-N-[6-(4-cyanophenyl)-6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 32)

步骤1：(4-氰基苯基)溴化镁的合成Step 1: Synthesis of (4-cyanophenyl)magnesium bromide

将4-溴苄腈(10.0g,54.94mmol,1.0eq)溶于超干的四氢呋喃(40.0mL)中，在氮气保护下，室温下滴加异丙基氯化镁氯化锂络合物(1.3mol/L,40.0mL)，常温反应16小时，得到棕色液体(4-氰基苯基)溴化镁的四氢呋喃溶液(0.66mol/L,82.26mL)。4-Bromobenzonitrile (10.0 g, 54.94 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (40.0 mL). Under nitrogen protection, isopropylmagnesium chloride lithium chloride complex (1.3 mol/L, 40.0 mL) was added dropwise at room temperature. The mixture was reacted at room temperature for 16 hours to obtain a brown liquid (4-cyanophenyl)magnesium bromide tetrahydrofuran solution (0.66 mol/L, 82.26 mL).

步骤2：4-(8-羟基-1,4-二氧螺[4.5]癸-8-基)苄腈的合成Step 2: Synthesis of 4-(8-hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)benzonitrile

将1,4-二氧阿司匹林[4.5]癸-8-酮(6.5g,41.67mmol,1.0eq)溶于超干的四氢呋喃(40.0mL)中，在氮气保护下，0℃滴加(4-氰基苯基)溴化镁的四氢呋喃溶液(75.0mL,1.2eq)，室温条件下反应3小时。反应完成后，在冰浴条件下，加入饱和氯化铵水溶液淬灭，再加水和乙酸乙酯萃取分液，所得有机相用饱和食盐水和无水硫酸钠干燥，过滤浓缩，得到黄色油状物4-(8-羟基-1,4-二氧螺[4.5]癸-8-基)苄腈粗品(3.1g,收率28.73％)。1H NMR(400MHz,DMSO)δ7.77(d,J＝8.2Hz,2H),7.65(d,J＝8.4Hz,2H),5.18(s,1H),3.89(d,J＝1.5Hz,4H),2.59(dd,J＝13.1,5.6Hz,2H),2.49–2.42(m,2H),2.22(dt,J＝13.9,5.2Hz,2H),2.14(dd,J＝13.1,5.6Hz,2H)。1,4-Dioxaspirino[4.5]decan-8-one (6.5 g, 41.67 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (40.0 mL). Under nitrogen, a solution of (4-cyanophenyl)magnesium bromide in tetrahydrofuran (75.0 mL, 1.2 eq) was added dropwise at 0°C. The mixture was allowed to react at room temperature for 3 hours. After completion, the reaction was quenched with saturated aqueous ammonium chloride in an ice bath. The mixture was then extracted with water and ethyl acetate. The resulting organic phase was dried over saturated brine and anhydrous sodium sulfate, filtered, and concentrated to afford crude 4-(8-hydroxy-1,4-dioxaspiro[4.5]decan-8-yl)benzonitrile (3.1 g, 28.73% yield) as a yellow oil. 1H NMR (400MHz, DMSO) δ7.77(d,J＝8.2Hz,2H),7.65(d,J＝8.4Hz,2H),5.18(s,1H),3.89(d,J＝1.5Hz,4H),2.5 9(dd,J＝13.1,5.6Hz,2H),2.49–2.42(m,2H),2.22(dt,J＝13.9,5.2Hz,2H),2.14(dd,J＝13.1,5.6Hz,2H).

步骤3：4-(1-羟基-4-氧代环己基)苄腈的合成Step 3: Synthesis of 4-(1-hydroxy-4-oxocyclohexyl)benzonitrile

将4-(8-羟基-1,4-二氧螺[4.5]癸-8-基)苄腈(2.74g,10.58mmol,1.0eq)溶于丙酮(10.0mL)中，随后加入对甲苯磺酸一水合物(604mg,3.17mmol,0.3eq)和水(20.0mL)，60℃反应2小时。反应完成后，加水和乙酸乙酯萃取分液，所得有机相用饱和食盐水和无水硫酸钠干燥，过滤浓缩，得到棕色固体4-(1-羟基-4-氧代环己基)苄腈粗品(1.07g,收率47.14％)。1H NMR(400MHz,DMSO)δ7.80(d,J＝8.4Hz,2H),7.76(d,J＝8.4Hz,2H),5.65(s,1H),2.75(m,J＝17.7,11.0,5.6Hz,3H),2.66–2.53(m,J＝4.1,3.5Hz,1H),2.28(td,J＝13.4,5.0Hz,2H),2.20–2.11(m,2H)。4-(8-Hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)benzonitrile (2.74 g, 10.58 mmol, 1.0 eq) was dissolved in acetone (10.0 mL), followed by the addition of p-toluenesulfonic acid monohydrate (604 mg, 3.17 mmol, 0.3 eq) and water (20.0 mL). The mixture was allowed to react at 60°C for 2 hours. After completion, the mixture was extracted with water and ethyl acetate. The resulting organic phase was dried over saturated brine and anhydrous sodium sulfate, filtered, and concentrated to afford crude 4-(1-hydroxy-4-oxocyclohexyl)benzonitrile (1.07 g, 47.14% yield) as a brown solid. 1H NMR (400MHz, DMSO) δ7.80(d,J＝8.4Hz,2H),7.76(d,J＝8.4Hz,2H),5.65(s,1H),2.75(m,J＝17.7,11 .0,5.6Hz,3H),2.66–2.53(m,J＝4.1,3.5Hz,1H),2.28(td,J＝13.4,5.0Hz,2H),2.20–2.11(m,2H).

步骤4：4-(2-氨基-6-羟基-4,5,6,7-四氢苯并[d]噻唑-6-基)苄腈的合成Step 4: Synthesis of 4-(2-amino-6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)benzonitrile

将4-(1-羟基-4-氧代环己基)苄腈(1.07g,4.98mmol,1.0eq)溶于超干的四氢呋喃(10.0mL)中，在冰浴条件下，逐滴加入苯基三甲基三溴化铵(1.87g,4.98mmol,1.0eq)的超干四氢呋喃溶液(8mL)，常温反应16小时。反应完成后，向反应液中加入水和乙酸乙酯萃取分液，浓缩有机相，所得产物溶于乙腈(20mL)中，再加入硫脲(568mg,7.47mmol,1.5eq)，在室温下反应4小时。反应完成后，过滤反应液，得到灰色固体4-(2-氨基-6-羟基-4,5,6,7-四氢苯并[d]噻唑-6-基)苄腈(210mg,收率15.56％)。LCMS(ESI)[M+H]+:272。1H NMR(400MHz,DMSO)δ9.20(s,2H),7.87–7.80(m,2H),7.76–7.69(m,2H),2.94(dd,J＝17.3,2.6Hz,1H),2.64(d,J＝16.5Hz,2H),2.52(s,1H),2.19(ddd,J＝13.0,10.7,6.0Hz,1H),1.91–1.80(m,1H)。4-(1-Hydroxy-4-oxocyclohexyl)benzonitrile (1.07 g, 4.98 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (10.0 mL). Phenyltrimethylammonium tribromide (1.87 g, 4.98 mmol, 1.0 eq) in ultra-dry tetrahydrofuran (8 mL) was added dropwise in an ice bath. The mixture was allowed to react at room temperature for 16 hours. After completion, water and ethyl acetate were added to the reaction mixture, and the organic phase was concentrated. The resulting product was dissolved in acetonitrile (20 mL), and thiourea (568 mg, 7.47 mmol, 1.5 eq) was added. The reaction was allowed to react at room temperature for 4 hours. After completion, the reaction mixture was filtered to obtain 4-(2-amino-6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)benzonitrile (210 mg, 15.56% yield) as a gray solid. LCMS (ESI) [M+H]+: 272. 1H NMR (400MHz, DMSO) δ9.20 (s, 2H), 7.87–7.80 (m, 2H), 7.76–7.69 (m, 2H), 2.94 (dd, J = 17.3, 2.6Hz, 1H), 2.64 (d, J＝16.5Hz, 2H), 2.52 (s, 1H), 2.19 (ddd, J＝13.0, 10.7, 6.0Hz, 1H), 1.91–1.80 (m, 1H).

步骤5：2'-氯-N-[6-(4-氰基苯基)-6-羟基-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-N-[6-(4-cyanophenyl)-6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(103mg,0.37mmol,1.0eq)和4-(2-氨基-6-羟基-4,5,6,7-四氢苯并[d]噻唑-6-基)苄腈(100mg,0.37mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(2.0mL)中，再加入N-甲基咪唑(121mg,1.48mmol,4.0eq)，随后在冰浴条件下，加入N,N,N′,N′-四甲基氯甲脒六氟磷酸盐(155mg,0.55mmol,1.5eq)，室温下反应16小时。反应结束后，制备分离纯化得到2'-氯-N-[6-(4-氰基苯基)-6-羟基-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(41mg,收率20.92％)。LCMS(ESI)[M+H]+:532.1210。1H NMR(400MHz,DMSO)δ12.50(s,1H),8.77(s,1H),8.17(s,1H),7.81(d,J＝8.5Hz,2H),7.73(d,J＝8.4Hz,2H),7.51(s,1H),7.41(s,1H),5.50(s,1H),3.62(s,3H),3.12(d,J＝16.4Hz,1H),2.79(d,J＝16.4Hz,2H),2.58(s,4H),2.30–2.18(m,1H),1.90(d,J＝12.8Hz,1H)。2'-Chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (103 mg, 0.37 mmol, 1.0 eq) and 4-(2-amino-6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)benzonitrile (100 mg, 0.37 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (2.0 mL). N-methylimidazole (121 mg, 1.48 mmol, 4.0 eq) was added, followed by N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (155 mg, 0.55 mmol, 1.5 eq) in an ice bath. The mixture was reacted at room temperature for 16 hours. After the reaction, preparative separation and purification were performed to obtain 2'-chloro-N-[6-(4-cyanophenyl)-6-hydroxy-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (41 mg, yield 20.92%). LCMS (ESI) [M+H]+: 532.1210. 1H NMR (400MHz, DMSO) δ12.50(s,1H),8.77(s,1H),8.17(s,1H),7.81(d,J＝8.5Hz,2H),7.73(d,J＝8.4Hz,2H),7.51(s,1H),7.41(s,1H) ,5.50(s,1H),3.62(s,3H),3.12(d,J＝16.4Hz,1H),2.79(d,J＝16.4Hz,2H),2.58(s,4H),2.30–2.18(m,1H),1.90(d,J＝12.8Hz,1H).

实施例25：2'-氯-N-{6-[4-(二氟甲氧基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物35)的合成
Example 25: Synthesis of 2'-chloro-N-{6-[4-(difluoromethoxy)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 35)

步骤1：8-[4-(二氟甲氧基)苯基]-1,4-二氧杂螺[4.5]癸-7-烯的合成Step 1: Synthesis of 8-[4-(difluoromethoxy)phenyl]-1,4-dioxaspiro[4.5]dec-7-ene

将(4-氰基-3-甲氧基苯基)硼酸(2.8g,16.0mmol,1.0eq)溶于1,4-二氧六环(20mL)中，随后依次加入1,4-二氧阿司匹林[4.5]癸-7-烯-8-基三氟甲磺酸酯(5.07g,17.6mmol,1.1eq)、碳酸铯(15.6g,48.0mmol,3.0eq)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(350mg,0.48mmol,0.03eq)和去离子水(5mL)，氮气保护下，80℃反应4小时。待反应完全，经正相(石油醚:乙酸乙酯＝11:1)纯化得白色固体8-[4-(二氟甲氧基)苯基]-1,4-二氧杂螺[4.5]癸-7-烯(3.1g,收率68.88％)。LCMS(ESI)[M+H]+:283.11。1H NMR(400MHz,DMSO-d6)δ7.49-7.43(m,2H),7.41-7.02(m,3H),6.00(m,J＝3.9,2.0Hz,1H),3.91(s,4H),2.57–2.50(m,2H),2.36(m,J＝4.3,2.4Hz,2H),1.81(t,J＝6.5Hz,2H)。(4-Cyano-3-methoxyphenyl)boronic acid (2.8 g, 16.0 mmol, 1.0 eq) was dissolved in 1,4-dioxane (20 mL), and then 1,4-dioxyaspirin[4.5]dec-7-en-8-yl trifluoromethanesulfonate (5.07 g, 17.6 mmol, 1.1 eq), cesium carbonate (15.6 g, 48.0 mmol, 3.0 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (350 mg, 0.48 mmol, 0.03 eq) and deionized water (5 mL) were added in sequence. Under nitrogen protection, the reaction was carried out at 80 ° C for 4 hours. After completion of the reaction, the product was purified by normal phase chromatography (petroleum ether:ethyl acetate = 11:1) to afford 8-[4-(difluoromethoxy)phenyl]-1,4-dioxaspiro[4.5]dec-7-ene (3.1 g, 68.88% yield) as a white solid. LCMS (ESI) [M + H] + : 283.11. 1 H NMR (400 MHz, DMSO-d6) δ 7.49-7.43 (m, 2H), 7.41-7.02 (m, 3H), 6.00 (m, J = 3.9, 2.0 Hz, 1H), 3.91 (s, 4H), 2.57–2.50 (m, 2H), 2.36 (m, J = 4.3, 2.4 Hz, 2H), 1.81 (t, J = 6.5 Hz, 2H).

步骤2：8-[4-(二氟甲氧基)苯基]-1,4-二氧杂螺[4.5]癸烷的合成Step 2: Synthesis of 8-[4-(difluoromethoxy)phenyl]-1,4-dioxaspiro[4.5]decane

将8-[4-(二氟甲氧基)苯基]-1,4-二氧杂螺[4.5]癸-7-烯(3.1g,10.99mmol,1.0eq)溶甲醇(20mL)中，加入钯炭(117.0mg,1.1mmol,0.1eq)，随后氢气氛围下，室温反应8小时。待反应完全，将反应液过滤，滤液浓缩后经正相(石油醚:乙酸乙酯＝21:1)纯化得白色固体8-[4-(二氟甲氧基)苯基]-1,4-二氧杂螺[4.5]癸烷(2.67g,收率85.5％)。LCMS(ESI)[M+H]+:285.13。1H NMR(400MHz,DMSO-d6)δ7.29-7.24(m,3H),7.08(d,J＝8.5Hz,2H),3.88(d,J＝2.1Hz,4H),2.59(m,J＝14.4,9.4,3.2Hz,1H),1.79-1.71(m,4H),1.70-1.58(m,4H)。8-[4-(Difluoromethoxy)phenyl]-1,4-dioxaspiro[4.5]dec-7-ene (3.1 g, 10.99 mmol, 1.0 eq) was dissolved in methanol (20 mL) and palladium on carbon (117.0 mg, 1.1 mmol, 0.1 eq) was added. The mixture was then reacted at room temperature under a hydrogen atmosphere for 8 hours. Upon completion, the reaction solution was filtered, concentrated, and purified via normal phase chromatography (petroleum ether:ethyl acetate = 21:1) to afford 8-[4-(difluoromethoxy)phenyl]-1,4-dioxaspiro[4.5]decane (2.67 g, 85.5% yield) as a white solid. LCMS (ESI) [M + H] + : 285.13. 1H NMR (400MHz, DMSO-d6) δ7.29-7.24(m,3H),7.08(d,J＝8.5Hz,2H),3.88(d,J＝2.1 Hz, 4H), 2.59 (m, J = 14.4, 9.4, 3.2Hz, 1H), 1.79-1.71 (m, 4H), 1.70-1.58 (m, 4H).

步骤3：4-[4-(二氟甲氧基)苯基]环己-1-酮的合成Step 3: Synthesis of 4-[4-(difluoromethoxy)phenyl]cyclohexan-1-one

将8-[4-(二氟甲氧基)苯基]-1,4-二氧杂螺[4.5]癸烷(1.0g,3.52mmol,1.0eq)溶丙酮/水(6mL/3mL)中，加入对甲苯磺酸-水合物(201mg,1.05mmol,0.3eq)，随后60℃反应2小时。待反应完全，加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯萃取，浓缩有机相得白色固体4-[4-(二氟甲氧基)苯基]环己-1-酮(810mg,收率95.88％)，直接用于下一步反应。LCMS(ESI)[M+H]+:241.10。1H NMR(400MHz,DMSO-d6)δ7.36(dd,J＝8.6,2.1Hz,2H),7.30-6.95(m,3H),3.07(m,J＝12.1,3.4Hz,1H),2.57(m,J＝14.1,6.0Hz,2H),2.26(m,J＝14.6,2.1Hz,2H),2.05(m,J＝12.4,6.0,2.8Hz,2H),1.86(m,J＝13.2,4.0Hz,2H)。8-[4-(Difluoromethoxy)phenyl]-1,4-dioxaspiro[4.5]decane (1.0 g, 3.52 mmol, 1.0 eq) was dissolved in acetone/water (6 mL/3 mL), and p-toluenesulfonic acid monohydrate (201 mg, 1.05 mmol, 0.3 eq) was added. The mixture was then reacted at 60°C for 2 hours. Upon completion, the pH was adjusted to neutral by adding saturated aqueous sodium bicarbonate solution. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to obtain 4-[4-(difluoromethoxy)phenyl]cyclohexan-1-one (810 mg, 95.88% yield) as a white solid, which was used directly in the next reaction. LCMS (ESI) [M + H] + : 241.10. 1H NMR (400MHz, DMSO-d6) δ7.36(dd,J＝8.6,2.1Hz,2H),7.30-6.95(m,3H),3.07(m,J＝12.1,3.4Hz,1H),2.57(m,J =14.1, 6.0Hz, 2H), 2.26 (m, J = 14.6, 2.1Hz, 2H), 2.05 (m, J = 12.4, 6.0, 2.8Hz, 2H), 1.86 (m, J = 13.2, 4.0Hz, 2H).

步骤4：6-[4-(二氟甲氧基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 4: Synthesis of 6-[4-(difluoromethoxy)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-[4-(二氟甲氧基)苯基]环己-1-酮(810mg,3.38mmol,1.0eq)溶于超干四氢呋喃(10mL)中，冰浴条件下，加入苯基三甲基三溴化铵(1.27g,3.38mmol,1.0eq)，常温反应2小时。待反应完全，加入水和乙酸乙酯萃取3次，浓缩干燥后再溶于乙腈(5mL)中，加入硫脲(385mg,5.06mmol,1.5eq)，常温反应2小时后，过滤，乙腈(20mL)清洗后得到白色固体6-[4-(二氟甲氧基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(251mg,收率25.63％)，直接用于下一步反应。LCMS(ESI)[M+H]+:297.08。4-[4-(Difluoromethoxy)phenyl]cyclohexan-1-one (810 mg, 3.38 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (10 mL). Phenyltrimethylammonium tribromide (1.27 g, 3.38 mmol, 1.0 eq) was added under ice-cooling and allowed to react at room temperature for 2 hours. After completion, the mixture was extracted three times with water and ethyl acetate. After concentration to dryness, the mixture was dissolved in acetonitrile (5 mL) and thiourea (385 mg, 5.06 mmol, 1.5 eq) was added. After reacting at room temperature for 2 hours, the mixture was filtered and washed with acetonitrile (20 mL) to obtain 6-[4-(difluoromethoxy)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (251 mg, 25.63% yield) as a white solid, which was used directly in the next step. LCMS (ESI) [M + H] + : 297.08.

步骤5：2'-氯-N-{6-[4-(二氟甲氧基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-N-{6-[4-(difluoromethoxy)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将6-[4-(二氟甲氧基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(248mg,0.84mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(232mg,0.84mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(5mL)中，加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(350mg,1.25mmol,1.5eq)和N-甲基咪唑(205mg,2.51mmol,3.0eq)，随后室温反应5.0小时。待反应完全，制备纯化得到白色固体2'-氯-N-{6-[4-(二氟甲氧基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(8.8mg,收率1.9％)。LCMS(ESI)[M+H]+:557.1229。1H NMR(400MHz,DMSO-d6)δ8.76(s,1H),8.16(d,J＝2.1Hz,1H),7.50(d,J＝2.1Hz,1H),7.39(d,J＝2.4Hz,2H),7.37(d,J＝2.1Hz,1H),7.18-6.96(m,3H),3.61(d,J＝2.0Hz,3H),3.02(d,J＝11.5Hz,1H),2.96-2.85(m,1H),2.71(t,J＝7.2Hz,3H),2.57(d,J＝2.1Hz,3H),1.99(d,J＝9.9Hz,2H)。6-[4-(Difluoromethoxy)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (248 mg, 0.84 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (232 mg, 0.84 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (5 mL), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (350 mg, 1.25 mmol, 1.5 eq) and N-methylimidazole (205 mg, 2.51 mmol, 3.0 eq) were added, followed by reaction at room temperature for 5.0 hours. After the reaction was complete, preparative purification was performed to obtain 2'-chloro-N-{6-[4-(difluoromethoxy)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (8.8 mg, 1.9% yield) as a white solid. LCMS (ESI) [M + H] + : 557.1229. 1H NMR (400MHz, DMSO-d6) δ8.76(s,1H),8.16(d,J＝2.1Hz,1H),7.50(d,J＝2.1Hz,1H),7.39(d,J＝2.4Hz,2H),7.37(d,J＝2.1Hz,1H),7.18-6.96( m, 3H), 3.61 (d, J = 2.0Hz, 3H), 3.02 (d, J = 11.5Hz, 1H), 2.96-2.85 (m, 1H), 2.71 (t, J = 7.2Hz, 3H), 2.57 (d, J = 2.1Hz, 3H), 1.99 (d, J = 9.9Hz, 2H).

实施例26：2'-氯-N-[6-(4-氰基-2-甲氧基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物41)的合成
Example 26: Synthesis of 2'-chloro-N-[6-(4-cyano-2-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 41)

步骤1：3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲腈的合成Step 1: Synthesis of 3-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzonitrile

将4,4,5,5-四甲基-2-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)-1,3,2-二氧杂硼烷(1.5g,5.63mmol,1.0eq)和4-溴-3-甲氧基苯甲腈(1.19g,5.63mmol,1.0eq)溶于1,4-二氧六环(20.0mL)和水(5.0mL)的混合溶液中，加入碳酸铯(5.49g,16.89mmol,3.0eq)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(124.27mg,0.17mmol,0.03eq)，在氮气氛围下，80℃反应2小时。待反应完全，正相纯化(石油醚/乙酸乙酯＝3:1)浓缩得白色固体3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲腈(1.3g,收率85.52％)。LCMS(ESI)[M+H]+:272.1。1H NMR(400MHz,DMSO)δ7.43(d,J＝1.5Hz,1H),7.36(dd,J＝7.7,1.5Hz,1H),7.26(d,J＝7.8Hz,1H),5.70(m,J＝3.8,1.9Hz,1H),3.91(s,4H),3.82(s,3H),2.52–2.43(m,2H),2.34(m,J＝4.2,2.3Hz,2H),1.75(t,J＝6.4Hz,2H)。4,4,5,5-Tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (1.5 g, 5.63 mmol, 1.0 eq) and 4-bromo-3-methoxybenzonitrile (1.19 g, 5.63 mmol, 1.0 eq) were dissolved in a mixed solution of 1,4-dioxane (20.0 mL) and water (5.0 mL). Cesium carbonate (5.49 g, 16.89 mmol, 3.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (124.27 mg, 0.17 mmol, 0.03 eq) were added, and the mixture was reacted at 80°C under a nitrogen atmosphere for 2 hours. After the reaction was complete, the product was purified by normal phase purification (petroleum ether/ethyl acetate = 3:1) and concentrated to give 3-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzonitrile (1.3 g, yield 85.52%) as a white solid. LCMS (ESI) [M + H] + : 272.1. 1H NMR (400MHz, DMSO) δ7.43(d,J＝1.5Hz,1H),7.36(dd,J＝7.7,1.5Hz,1H),7.26(d,J＝7.8Hz,1H),5.70(m,J＝3.8 ,1.9Hz,1H),3.91(s,4H),3.82(s,3H),2.52–2.43(m,2H),2.34(m,J＝4.2,2.3Hz,2H),1.75(t,J＝6.4Hz,2H).

步骤2：3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-8-基)苯甲腈的合成Step 2: Synthesis of 3-methoxy-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzonitrile

将3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲腈(1.3g,4.79mmol,1.0eq)溶甲醇(30mL)中，加入钯炭(50.9mg,0.48mmol,0.1eq)，随后氢气氛围下，反应8小时。待反应完全，将反应液过滤，滤液浓缩后，经正相(石油醚:乙酸乙酯＝20:1)纯化得到白色固体3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-8-基)苯甲腈(220mg,收率16.8％)。LCMS(ESI)[M+H]+:274.14。1H NMR(400MHz,DMSO)δ7.40(d,J＝1.3Hz,1H),7.36–7.34(m,2H),3.87(d,J＝1.8Hz,4H),3.85(s,3H),3.01–2.91(m,1H),1.77–1.56(m,8H)。3-Methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzonitrile (1.3 g, 4.79 mmol, 1.0 eq) was dissolved in methanol (30 mL) and palladium on carbon (50.9 mg, 0.48 mmol, 0.1 eq) was added. The mixture was then reacted under a hydrogen atmosphere for 8 hours. Upon completion, the reaction solution was filtered, concentrated, and purified via normal phase chromatography (petroleum ether:ethyl acetate = 20:1) to afford 3-methoxy-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzonitrile (220 mg, 16.8% yield) as a white solid. LCMS (ESI) [M + H] + : 274.14. 1 H NMR (400MHz, DMSO) δ7.40 (d, J = 1.3 Hz, 1H), 7.36–7.34 (m, 2H), 3.87 (d, J = 1.8 Hz, 4H), 3.85 (s, 3H), 3.01–2.91 (m, 1H), 1.77–1.56 (m, 8H).

步骤3：3-甲氧基-4-(4-氧代环己基)苯甲腈的合成Step 3: Synthesis of 3-methoxy-4-(4-oxocyclohexyl)benzonitrile

将3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-8-基)苯甲腈(220mg,0.80mmol,1.0eq)溶丙酮/水(8mL/4mL)中，加对甲苯磺酸-水合物(45.9mg,0.24mmol,0.3eq)，随后60℃反应2小时。待反应完全，加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯萃取，浓缩有机相得白色固体3-甲氧基-4-(4-氧代环己基)苯甲腈(165mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:230.11。1H NMR(400MHz,DMSO)δ7.46–7.34(m,3H),3.89(s,3H),3.45(m,J＝12.1,3.4Hz,1H),2.59(m,J＝14.2,6.0Hz,2H),2.26(m,J＝14.7,4.4,2.1Hz,2H),2.00(m,J＝12.6,6.0,3.0Hz,2H),1.85(m,J＝13.1,4.1Hz,2H)。3-Methoxy-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzonitrile (220 mg, 0.80 mmol, 1.0 eq) was dissolved in acetone/water (8 mL/4 mL) and p-toluenesulfonic acid monohydrate (45.9 mg, 0.24 mmol, 0.3 eq) was added. The mixture was reacted at 60°C for 2 hours. Upon completion, the pH was adjusted to neutral by adding saturated aqueous sodium bicarbonate solution. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to obtain 3-methoxy-4-(4-oxocyclohexyl)benzonitrile (165 mg) as a white solid, which was used directly in the next reaction. LCMS (ESI) [M + H] + : 230.11. 1H NMR (400MHz, DMSO) δ7.46–7.34(m,3H),3.89(s,3H),3.45(m,J＝12.1,3.4Hz,1H),2.59(m,J＝14.2,6.0H z, 2H), 2.26 (m, J = 14.7, 4.4, 2.1Hz, 2H), 2.00 (m, J = 12.6, 6.0, 3.0Hz, 2H), 1.85 (m, J = 13.1, 4.1Hz, 2H).

步骤4：4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-3-甲氧基苯甲腈的合成Step 4: Synthesis of 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-3-methoxybenzonitrile

将3-甲氧基-4-(4-氧代环己基)苯甲腈(150mg,0.65mmol,1.0eq)溶于超干四氢呋喃(10mL)中，冰浴条件下，加入苯基三甲基三溴化铵(245.1mg,0.65mmol,1.0eq)，浓缩干燥后再溶于乙腈(3mL)中，加入硫脲(54.6mg,0.72mmol,1.1eq)，常温反应16小时。待反应结束，将反应液过滤，乙腈(20mL)清洗后得到白色固体粗品4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-3-甲氧基苯甲腈(197mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:286。3-Methoxy-4-(4-oxocyclohexyl)benzonitrile (150 mg, 0.65 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (10 mL). Phenyltrimethylammonium tribromide (245.1 mg, 0.65 mmol, 1.0 eq) was added under ice-cooling. The mixture was concentrated to dryness and then dissolved in acetonitrile (3 mL). Thiourea (54.6 mg, 0.72 mmol, 1.1 eq) was added and allowed to react at room temperature for 16 hours. Upon completion of the reaction, the reaction mixture was filtered and washed with acetonitrile (20 mL) to obtain a crude white solid, 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-3-methoxybenzonitrile (197 mg), which was used directly in the next step. LCMS (ESI) [M + H] + : 286.

步骤5：2'-氯-N-[6-(4-氰基-2-甲氧基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-N-[6-(4-cyano-2-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-3-甲氧基苯甲腈(102.6mg,0.36mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(100mg,0.36mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(2mL)中，加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(151.4mg,0.54mmol,1.5eq)和N-甲基咪唑(88.56mg,1.08mmol,3.0eq)，随后室温反应1.5小时。待反应完全，制备得白色固体2'-氯-N-[6-(4-氰基-2-甲氧基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(14.4mg,收率7.34％)。LCMS(ESI)[M+H]+:546.1362。1H NMR(400MHz,DMSO)δ12.49(s,1H),8.77(s,1H),8.17(s,1H),7.51(s,1H),7.47–7.38(m,4H),3.87(s,3H),3.62(s,3H),3.45–3.35(m,1H),2.89(dd,J＝15.6,4.9Hz,1H),2.70(m,J＝9.5Hz,3H),2.58(s,3H),2.14–1.91(m,2H)。4-(2-Amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-3-methoxybenzonitrile (102.6 mg, 0.36 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (100 mg, 0.36 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (2 mL), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (151.4 mg, 0.54 mmol, 1.5 eq) and N-methylimidazole (88.56 mg, 1.08 mmol, 3.0 eq) were added, followed by reaction at room temperature for 1.5 hours. After the reaction was complete, a white solid, 2'-chloro-N-[6-(4-cyano-2-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (14.4 mg, 7.34% yield) was obtained. LCMS (ESI) [M + H]+: 546.1362. 1H NMR(400MHz,DMSO)δ12.49(s,1H),8.77(s,1H),8.17(s,1H),7.51(s,1H),7.47–7.38(m,4H),3.87(s,3H),3.62 (s,3H),3.45–3.35(m,1H),2.89(dd,J＝15.6,4.9Hz,1H),2.70(m,J＝9.5Hz,3H),2.58(s,3H),2.14–1.91(m,2H).

实施例27：2'-氯-N-{6-[4-(二氟甲基)-2-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物42)的合成
Example 27: Synthesis of 2'-chloro-N-{6-[4-(difluoromethyl)-2-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 42)

步骤1：3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲醛的合成Step 1: Synthesis of 3-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzaldehyde

将3-甲氧基-4溴-苯甲酰(1.0g,4.67mmol,1.0eq)和4,4,5,5-四甲基-2-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)-1,3,2-二氧杂硼烷(1.24g,4.67mmol,1.0eq)溶于1,4-二氧六环(12mL)中，加入碳酸铯(4.56g,14.01mmol,3.0eq)、去离子水(3mL)和1,1'-双二苯基膦二茂铁二氯化钯(170mg,0.23mmol,0.05eq)，氮气保护下，80℃反应4小时。待反应完成后，加入去离子水和乙酸乙酯萃取，浓缩有机相得到棕色固体3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲醛(1.27g)，未经进一步纯化，直接用于下一步反应。LCMS(ESI)[M+H]+:275。3-Methoxy-4-bromo-benzoyl (1.0 g, 4.67 mmol, 1.0 eq) and 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (1.24 g, 4.67 mmol, 1.0 eq) were dissolved in 1,4-dioxane (12 mL), and cesium carbonate (4.56 g, 14.01 mmol, 3.0 eq), deionized water (3 mL) and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (170 mg, 0.23 mmol, 0.05 eq) were added. The mixture was reacted at 80 ° C for 4 hours under nitrogen protection. After the reaction was complete, deionized water and ethyl acetate were added for extraction, and the organic phase was concentrated to obtain a brown solid of 3-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzaldehyde (1.27 g). This was used directly in the next reaction without further purification. LCMS (ESI) [M+H] + : 275.

步骤2：8-[4-(二氟甲基)-2-甲氧基苯基]-1,4-二氧杂螺[4.5]癸-7-烯的合成Step 2: Synthesis of 8-[4-(difluoromethyl)-2-methoxyphenyl]-1,4-dioxaspiro[4.5]dec-7-ene

将3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲醛(800mg,2.55mmol,1.0eq)溶于超干二氯甲烷(10mL)中，冰浴条件下，缓慢加入二乙胺基三氟化硫(617mg,3.83mmol,1.5eq)，随后恢复至室温条件下反应3小时。待反应完全后，加入饱和碳酸氢钠溶液淬灭，加入乙酸乙酯萃取，浓缩有机相，经正相柱层析纯化(石油醚：乙酸乙酯＝90：10)得黄色油状物8-[4-(二氟甲基)-2-甲氧基苯基]-1,4-二氧杂螺[4.5]癸-7-烯(610mg,收率70.6％)。LCMS(ESI)[M+H]+:297。1H NMR(400MHz,DMSO)δ7.21(d,J＝7.6Hz,1H),7.14(s,1H),7.09(d,J＝7.8Hz,1H),7.12–6.82(m,1H),5.73–5.56(m,1H),3.92(s,4H),3.80(s,3H),2.49–2.45(m,2H),2.33(m,J＝2.7Hz,2H),1.75(t,J＝6.4Hz,2H)。3-Methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzaldehyde (800 mg, 2.55 mmol, 1.0 eq) was dissolved in ultra-dry dichloromethane (10 mL). Diethylaminosulfur trifluoride (617 mg, 3.83 mmol, 1.5 eq) was slowly added under an ice bath. The mixture was then allowed to react at room temperature for 3 hours. After completion, the reaction was quenched with saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was concentrated and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 90:10) to afford 8-[4-(difluoromethyl)-2-methoxyphenyl]-1,4-dioxaspiro[4.5]dec-7-ene (610 mg, 70.6% yield) as a yellow oil. LCMS (ESI) [M+H] + : 297. 1H NMR (400MHz, DMSO) δ7.21(d,J＝7.6Hz,1H),7.14(s,1H),7.09(d,J＝7.8Hz,1H),7.12–6.82(m,1H),5.73– 5.56(m,1H),3.92(s,4H),3.80(s,3H),2.49–2.45(m,2H),2.33(m,J＝2.7Hz,2H),1.75(t,J＝6.4Hz,2H).

步骤3：8-[4-(二氟甲基)-2-甲氧基苯基]-1,4-二氧杂螺[4.5]癸烷的合成Step 3: Synthesis of 8-[4-(difluoromethyl)-2-methoxyphenyl]-1,4-dioxaspiro[4.5]decane

将8-[4-(二氟甲基)-2-甲氧基苯基]-1,4-二氧杂螺[4.5]癸-7-烯(580mg,1.96mmol,1.0eq)溶于无水甲醇(10mL)中，加入钯碳(58mg,10％)，在氢气氛围下，常温常压下反应2小时。待反应完全后，将反应液过滤，滤液浓缩得到黄色油状物8-[4-(二氟甲基)-2-甲氧基苯基]-1,4-二氧杂螺[4.5]癸烷(582mg,收率99.8％)，直接用于下一步反应。LCMS(ESI)[M+H]+:299。1H NMR(400MHz,DMSO)δ7.29(d,J＝7.6Hz,1H),7.10(d,J＝7.7Hz,2H),7.09–6.80(m,1H),3.88(d,J＝2.3Hz,4H),3.83(s,3H),3.02–2.88(m,1H),1.79–1.55(m,8H)。8-[4-(Difluoromethyl)-2-methoxyphenyl]-1,4-dioxaspiro[4.5]dec-7-ene (580 mg, 1.96 mmol, 1.0 eq) was dissolved in anhydrous methanol (10 mL), and palladium on carbon (58 mg, 10%) was added. The mixture was reacted under a hydrogen atmosphere at room temperature and pressure for 2 hours. After the reaction was complete, the reaction solution was filtered and the filtrate was concentrated to obtain a yellow oil, 8-[4-(difluoromethyl)-2-methoxyphenyl]-1,4-dioxaspiro[4.5]decane (582 mg, 99.8% yield), which was used directly in the next reaction. LCMS (ESI) [M+H] + : 299. 1H NMR (400MHz, DMSO) δ7.29(d,J＝7.6Hz,1H),7.10(d,J＝7.7Hz,2H),7.09–6.80(m ,1H),3.88(d,J＝2.3Hz,4H),3.83(s,3H),3.02–2.88(m,1H),1.79–1.55(m,8H).

步骤4：4-[4-(二氟甲基)-2-甲氧基苯基]环己-1-酮的合成Step 4: Synthesis of 4-[4-(difluoromethyl)-2-methoxyphenyl]cyclohexan-1-one

将8-[4-(二氟甲基)-2-甲氧基苯基]-1,4-二氧杂螺[4.5]癸烷(530mg,1.78mmol,1.0eq)溶于丙酮(5mL)与水(5mL)的混合溶剂中，加入对甲苯磺酸一水合物(101mg,0.53mmol,0.3eq)，60℃反应6小时。待反应完全后，加入乙酸乙酯萃取，浓缩有机相得到黄色油状物4-[4-(二氟甲基)-2-甲氧基苯基]环己-1-酮(380mg,收率84.3％)。LCMS(ESI)[M+H]+:255。1H NMR(400MHz,DMSO)δ7.35(d,J＝7.8Hz,1H),7.16(s,1H),7.11(s,1H),7.12–6.82(m,1H),3.87(s,3H),3.43(m,J＝10.6,3.7Hz,1H),2.59(td,J＝14.2,6.0Hz,2H),2.27(m,J＝14.2,4.4,2.1Hz,2H),2.02(m,J＝11.4,4.6,2.5Hz,2H),1.85(m,J＝13.1,4.1Hz,2H)。8-[4-(Difluoromethyl)-2-methoxyphenyl]-1,4-dioxaspiro[4.5]decane (530 mg, 1.78 mmol, 1.0 eq) was dissolved in a mixture of acetone (5 mL) and water (5 mL). p-Toluenesulfonic acid monohydrate (101 mg, 0.53 mmol, 0.3 eq) was added and reacted at 60°C for 6 hours. After the reaction was complete, ethyl acetate was added for extraction, and the organic phase was concentrated to obtain 4-[4-(difluoromethyl)-2-methoxyphenyl]cyclohexan-1-one (380 mg, 84.3% yield) as a yellow oil. LCMS (ESI) [M+H] + : 255. 1H NMR (400MHz, DMSO) δ7.35(d,J＝7.8Hz,1H),7.16(s,1H),7.11(s,1H),7.12–6.82(m,1H),3.87(s,3H),3.43(m,J＝10.6,3.7Hz,1H ), 2.59(td,J＝14.2,6.0Hz,2H), 2.27(m,J＝14.2,4.4,2.1Hz,2H), 2.02(m,J＝11.4,4.6,2.5Hz,2H), 1.85(m,J＝13.1,4.1Hz,2H).

步骤5：6-[4-(二氟甲基)-2-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 5: Synthesis of 6-[4-(difluoromethyl)-2-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-[4-(二氟甲基)-2-甲氧基苯基]环己-1-酮(300mg,1.18mmol,1.0eq)溶于超干四氢呋喃(5mL)中，分批加苯基三甲基三溴化铵(488mg,1.30mmol,1.1eq)，在常温常压下反应1小时。待反应完全后，用乙酸乙酯萃取，浓缩有机相，并用乙腈(3mL)重新溶解，随后加入硫脲(89.7mg,1.18mmol,1.0eq)，60℃反应3小时，待反应完全后，静置冷却至室温，过滤得到类白色固体6-[4-(二氟甲基)-2-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(135mg,收率36.9％)，未经进一步纯化，直接用于下一步反应。LCMS(ESI)[M+H]+:311。4-[4-(Difluoromethyl)-2-methoxyphenyl]cyclohexan-1-one (300 mg, 1.18 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (5 mL), and phenyltrimethylammonium tribromide (488 mg, 1.30 mmol, 1.1 eq) was added portionwise. The mixture was allowed to react at room temperature and pressure for 1 hour. After the reaction was complete, the mixture was extracted with ethyl acetate, and the organic phase was concentrated and redissolved in acetonitrile (3 mL). Thiourea (89.7 mg, 1.18 mmol, 1.0 eq) was then added. The mixture was allowed to react at 60°C for 3 hours. After the reaction was complete, the mixture was allowed to cool to room temperature and filtered to afford 6-[4-(difluoromethyl)-2-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (135 mg, 36.9% yield) as an off-white solid. This was used directly in the next step without further purification. LCMS (ESI) [M+H] + : 311.

步骤6：2'-氯-N-{6-[4-(二氟甲基)-2-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of 2'-chloro-N-{6-[4-(difluoromethyl)-2-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将6-[4-(二氟甲基)-2-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(100mg,0.32mmol,1.0eq)溶于乙腈(2mL)中，依次加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(90mg,0.32mmol,1.0eq)和N-甲基咪唑(79.2mg,0.97mmol,3.0eq)，在常温常压下反应12小时。待反应完全后，制备得到白色固体2'-氯-N-{6-[4-(二氟甲基)-2-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(4.1mg,收率2.34％)。LCMS(ESI)[M+H]+:571.14。1H NMR(400MHz,DMSO)δ12.50(s,1H),8.76(s,1H),8.16(s,1H),7.51(s,1H),7.44–7.35(m,2H),7.17–6.82(m,3H),3.84(s,3H),3.62(s,3H),2.89(dd,J＝15.6,4.8Hz,1H),2.76–2.63(m,3H),2.58(s,3H),2.55(s,1H),2.13–1.92(m,2H)。6-[4-(Difluoromethyl)-2-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (100 mg, 0.32 mmol, 1.0 eq) was dissolved in acetonitrile (2 mL). N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (90 mg, 0.32 mmol, 1.0 eq) and N-methylimidazole (79.2 mg, 0.97 mmol, 3.0 eq) were added sequentially. The mixture was allowed to react at room temperature and pressure for 12 hours. Upon completion of the reaction, 2'-chloro-N-{6-[4-(difluoromethyl)-2-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (4.1 mg, 2.34% yield) was obtained as a white solid. LCMS(ESI)[M+H] + :571.14. 1H NMR (400MHz, DMSO) δ12.50(s,1H),8.76(s,1H),8.16(s,1H),7.51(s,1H),7.44–7.35(m,2H),7.17–6.82(m,3H),3.8 4(s,3H),3.62(s,3H),2.89(dd,J＝15.6,4.8Hz,1H),2.76–2.63(m,3H),2.58(s,3H),2.55(s,1H),2.13–1.92(m,2H).

实施例28：2'-氯-N-[6-(4-氰基-3-甲氧基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物43)的合成
Example 28: Synthesis of 2'-chloro-N-[6-(4-cyano-3-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 43)

步骤1：2-甲氧基-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯甲腈的合成Step 1: Synthesis of 2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile

将4-溴-2-甲氧基苯甲腈(5.0g,23.69mmol,1.0eq)溶于超干1,4-二氧六环(50.0mL)中，加入联硼酸频那醇酯(11.98g,47.39mmol,2.0eq)、乙酸钾(6.96g,71.07mmol,3.0eq)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(517mg,0.71mmol,0.03eq)，在氮气保护下，100℃反应5小时。待反应完全，冷却至室温后过滤、浓缩得黑色油状物2-甲氧基-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯甲腈(5.8g,收率94.9％)，直接用于下一步反应。LCMS(ESI)[M+H]+:260.14。4-Bromo-2-methoxybenzonitrile (5.0 g, 23.69 mmol, 1.0 eq) was dissolved in ultra-dry 1,4-dioxane (50.0 mL). Diboronic acid pinacol diboron (11.98 g, 47.39 mmol, 2.0 eq), potassium acetate (6.96 g, 71.07 mmol, 3.0 eq), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (517 mg, 0.71 mmol, 0.03 eq) were added. The mixture was reacted at 100°C under nitrogen for 5 hours. After completion of the reaction, the mixture was cooled to room temperature, filtered, and concentrated to afford 2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (5.8 g, 94.9% yield) as a black oil, which was used directly in the next reaction. LCMS (ESI) [M + H] + : 260.14.

步骤2：2-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲腈的合成Step 2: Synthesis of 2-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzonitrile

将2-甲氧基-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯甲腈(5.8g,22.39mmol,1.0eq)溶于1,4-二氧六环(20mL)中，随后加入1,4-二氧阿司匹林[4.5]癸-7-烯-8-基三氟甲磺酸酯(6.45g,22.39mmol,1.0eq)、碳酸铯(21.83g,67.17mmol,3.0eq)、去离子水(5mL)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(489.77mg,0.67mmol,0.03eq)，氮气保护下，80℃反应4小时。待反应完全，经正相(石油醚:乙酸乙酯＝7:1)纯化得到白色油状物2-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲腈(4.8g,收率80.51％)。LCMS(ESI)[M+H]+:272.12。2-Methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (5.8 g, 22.39 mmol, 1.0 eq) was dissolved in 1,4-dioxane (20 mL), followed by the addition of 1,4-dioxyaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (6.45 g, 22.39 mmol, 1.0 eq), cesium carbonate (21.83 g, 67.17 mmol, 3.0 eq), deionized water (5 mL) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (489.77 mg, 0.67 mmol, 0.03 eq), and the reaction was carried out at 80 ° C for 4 hours under nitrogen protection. After the reaction was complete, the product was purified by normal phase chromatography (petroleum ether:ethyl acetate = 7:1) to give 2-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzonitrile (4.8 g, yield 80.51%) as a white oil. LCMS (ESI) [M + H] + : 272.12.

步骤3：2-甲氧基-4-(1,4-二氧杂螺[4.5]癸-8-基)苯甲腈的合成Step 3: Synthesis of 2-methoxy-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzonitrile

将2-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲腈(4.8g,17.71mmol,1.0eq)溶甲醇中(20mL)中，加入钯炭(187.7mg,1.77mmol,0.1eq)，随后氢气氛围下，室温反应8小时。待反应完全，将反应液过滤，滤液浓缩后经正相(石油醚:乙酸乙酯＝21:4)纯化得到白色固体2-甲氧基-4-(1,4-二氧杂螺[4.5]癸-8-基)苯甲腈(3.84g,收率79.5％)。LCMS(ESI)[M+H]+:274.14。1H NMR(400MHz,DMSO-d6)δ7.61(d,J＝7.9Hz,1H),7.06(d,J＝1.4Hz,1H),6.95(dd,J＝7.9,1.4Hz,1H),3.91(s,3H),3.89(d,J＝2.2Hz,4H),2.68(m,J＝11.3,3.4Hz,1H),1.77(d,J＝10.2Hz,4H),1.63(m,J＝23.7,11.6,3.2Hz,4H)。2-Methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzonitrile (4.8 g, 17.71 mmol, 1.0 eq) was dissolved in methanol (20 mL) and palladium on carbon (187.7 mg, 1.77 mmol, 0.1 eq) was added. The mixture was then reacted at room temperature under a hydrogen atmosphere for 8 hours. Upon completion, the reaction solution was filtered, concentrated, and purified via normal phase chromatography (petroleum ether:ethyl acetate = 21:4) to afford 2-methoxy-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzonitrile (3.84 g, 79.5% yield) as a white solid. LCMS (ESI) [M + H] + : 274.14. 1H NMR (400MHz, DMSO-d6) δ7.61(d,J＝7.9Hz,1H),7.06(d,J＝1.4Hz,1H),6.95(dd,J＝7.9,1.4Hz,1H),3.91(s,3H) ,3.89(d,J＝2.2Hz,4H),2.68(m,J＝11.3,3.4Hz,1H),1.77(d,J＝10.2Hz,4H),1.63(m,J＝23.7,11.6,3.2Hz,4H).

步骤4：2-甲氧基-4-(4-氧代环己基)苯甲腈的合成Step 4: Synthesis of 2-methoxy-4-(4-oxocyclohexyl)benzonitrile

将2-甲氧基-4-(1,4-二氧杂螺[4.5]癸-8-基)苯甲腈(1.0g,3.65mmol,1.0eq)溶丙酮/水(6mL/3mL)中，加对甲苯磺酸-水合物(208mg,1.09mmol,0.3eq)，随后60℃反应2.0小时。待反应完全，加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯萃取，浓缩有机相得白色固体2-甲氧基-4-(4-氧代环己基)苯甲腈(820mg,收率98.1％)，直接用于下一步反应。LCMS(ESI)[M+H]+:230.11。1H NMR(400MHz,DMSO-d6)δ7.64(d,J＝7.9Hz,1H),7.19(d,J＝1.5Hz,1H),7.04(dd,J＝7.9,1.4Hz,1H),3.92(s,3H),3.15(m,J＝12.0,3.5Hz,1H),2.63-2.52(m,2H),2.28(m,J＝14.6,4.4,2.1Hz,2H),2.07(m,J＝12.4,6.2,3.6Hz,2H),2.00-1.89(m,2H)。2-Methoxy-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzonitrile (1.0 g, 3.65 mmol, 1.0 eq) was dissolved in acetone/water (6 mL/3 mL) and p-toluenesulfonic acid monohydrate (208 mg, 1.09 mmol, 0.3 eq) was added. The mixture was then reacted at 60°C for 2.0 hours. Upon completion, the pH was adjusted to neutral by adding saturated aqueous sodium bicarbonate solution. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to obtain 2-methoxy-4-(4-oxocyclohexyl)benzonitrile (820 mg, 98.1% yield) as a white solid, which was used directly in the next reaction. LCMS (ESI) [M + H] + : 230.11. 1H NMR (400MHz, DMSO-d6) δ7.64(d,J＝7.9Hz,1H),7.19(d,J＝1.5Hz,1H),7.04(dd,J＝7.9,1.4Hz,1H),3.92(s,3H),3.15(m,J＝ 12.0,3.5Hz,1H),2.63-2.52(m,2H),2.28(m,J＝14.6,4.4,2.1Hz,2H),2.07(m,J＝12.4,6.2,3.6Hz,2H),2.00-1.89(m,2H).

步骤5：4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-2-甲氧基苯甲腈的合成Step 5: Synthesis of 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-2-methoxybenzonitrile

将2-甲氧基-4-(4-氧代环己基)苯甲腈(800mg,3.51mmol,1.0eq)溶于超干四氢呋喃(10mL)中，冰浴条件下，缓慢加入苯基三甲基三溴化铵(1.32g,3.51mmol,1.0eq)，室温反应2小时，加入水和乙酸乙酯萃取分液，干燥浓缩后再溶于乙腈(5mL)中，加入硫脲(320mg,4.21mmol,1.2eq)，常温反应2小时后，过滤，乙腈(20mL)清洗后，得白色固体4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-2-甲氧基苯甲腈(450mg,收率45.0％)。LCMS(ESI)[M+H]+:286.1。1H NMR(400MHz,DMSO-d6)δ9.22(s,2H),7.68(d,J＝8.0Hz,1H),7.22(d,J＝1.4Hz,1H),7.05(dd,J＝8.1,1.4Hz,1H),3.92(s,3H),3.10(m,J＝10.2,5.2Hz,1H),2.81(dd,J＝15.9,5.2Hz,1H),2.68(m,J＝13.9,3.0Hz,1H),2.61(d,J＝7.3Hz,2H),2.00(m,J＝4.9,3.3Hz,2H)。2-Methoxy-4-(4-oxocyclohexyl)benzonitrile (800 mg, 3.51 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (10 mL). Phenyltrimethylammonium tribromide (1.32 g, 3.51 mmol, 1.0 eq) was slowly added in an ice bath. The mixture was allowed to react at room temperature for 2 hours. Water and ethyl acetate were added for extraction, dried, concentrated, and then dissolved in acetonitrile (5 mL). Thiourea (320 mg, 4.21 mmol, 1.2 eq) was added and allowed to react at room temperature for 2 hours. The mixture was then filtered and washed with acetonitrile (20 mL) to obtain 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-2-methoxybenzonitrile (450 mg, 45.0% yield) as a white solid. LCMS (ESI) [M + H] + : 286.1. 1H NMR (400MHz, DMSO-d6) δ9.22(s,2H),7.68(d,J＝8.0Hz,1H),7.22(d,J＝1.4Hz,1H),7.05(dd,J＝8.1,1.4Hz,1H),3.92(s,3H),3.10( m,J＝10.2,5.2Hz,1H),2.81(dd,J＝15.9,5.2Hz,1H),2.68(m,J＝13.9,3.0Hz,1H),2.61(d,J＝7.3Hz,2H),2.00(m,J＝4.9,3.3Hz,2H).

步骤6：2'-氯-N-[6-(4-氰基-3-甲氧基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of 2'-chloro-N-[6-(4-cyano-3-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-2-甲氧基苯甲腈(200.0mg,0.70mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(195mg,0.70mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(3mL)中，加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(295mg,1.05mmol,1.5eq)和N-甲基咪唑(173mg,2.10mmol,3.0eq)，随后室温反应5小时。待反应完全，制备得白色固体2'-氯-N-[6-(4-氰基-3-甲氧基苯基)-4,5,6,7-四氢苯并[d]噻唑-2-基]-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(8.2mg,收率2.1％)。LCMS(ESI)[M+H]+:546.1370。1H NMR(400MHz,DMSO-d6)δ12.51(s,1H),8.77(s,1H),8.17(s,1H),7.66(d,J＝8.0Hz,1H),7.52(s,1H),7.41(s,1H),7.24(d,J＝1.4Hz,1H),7.07(dd,J＝8.1,1.4Hz,1H),3.92(s,3H),3.61(s,3H),3.12(dt,J＝10.5,5.6Hz,1H),3.01–2.77(m,2H),2.73(s,2H),2.58(s,3H),2.05(s,2H)。4-(2-Amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-2-methoxybenzonitrile (200.0 mg, 0.70 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (195 mg, 0.70 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (3 mL), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (295 mg, 1.05 mmol, 1.5 eq) and N-methylimidazole (173 mg, 2.10 mmol, 3.0 eq) were added, followed by reaction at room temperature for 5 hours. After the reaction was complete, a white solid, 2'-chloro-N-[6-(4-cyano-3-methoxyphenyl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl]-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (8.2 mg, 2.1% yield) was obtained. LCMS (ESI) [M + H] + : 546.1370. 1 H NMR (400MHz, DMSO-d 6 )δ12.51(s,1H),8.77(s,1H),8.17(s,1H),7.66(d,J＝8.0Hz,1H),7.52(s,1H),7.41(s,1H),7.24(d,J＝1.4Hz,1H),7.07(dd,J＝8. 1,1.4Hz,1H),3.92(s,3H),3.61(s,3H),3.12(dt,J＝10.5,5.6Hz,1H),3.01–2.77(m,2H),2.73(s,2H),2.58(s,3H),2.05(s,2H).

实施例29：2'-氯-N-{6-[4-(二氟甲基)-3-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物44)的合成
Example 29: Synthesis of 2'-chloro-N-{6-[4-(difluoromethyl)-3-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 44)

步骤1：3-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲醛的合成Step 1: Synthesis of 3-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzaldehyde

将2-甲氧基-4溴-苯甲酰(1.0g,4.67mmol,1.0eq)和4,4,5,5-四甲基-2-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)-1,3,2-二氧杂硼烷(1.24g,4.67mmol,1.0eq)溶于1,4-二氧六环(12mL)中，加入碳酸铯(4.56g,14.01mmol,3.0eq)、去离子水(3mL)和1,1'-双二苯基膦二茂铁二氯化钯(170mg,0.23mmol,0.05eq)，在氮气保护下，80℃反应4小时。待反应完成后，加入去离子水和乙酸乙酯萃取，浓缩有机相得棕色固体2-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲醛(1.26g,收率98.4％)，未经进一步纯化，直接用于下一步反应。LCMS(ESI)[M+H]+:275。2-Methoxy-4-bromo-benzoyl (1.0 g, 4.67 mmol, 1.0 eq) and 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (1.24 g, 4.67 mmol, 1.0 eq) were dissolved in 1,4-dioxane (12 mL), and cesium carbonate (4.56 g, 14.01 mmol, 3.0 eq), deionized water (3 mL) and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (170 mg, 0.23 mmol, 0.05 eq) were added. The mixture was reacted at 80 ° C for 4 hours under nitrogen protection. After the reaction was complete, deionized water and ethyl acetate were added for extraction. The organic phase was concentrated to obtain a brown solid of 2-methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzaldehyde (1.26 g, 98.4% yield). This was used directly in the next reaction without further purification. LCMS (ESI) [M+H] + : 275.

步骤2：8-[4-(二氟甲基)-3-甲氧基苯基]-1,4-二氧杂螺[4.5]癸-7-烯的合成Step 2: Synthesis of 8-[4-(difluoromethyl)-3-methoxyphenyl]-1,4-dioxaspiro[4.5]dec-7-ene

将2-甲氧基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯甲醛(1.0g,3.19mmol,1.0eq)溶于超干二氯甲烷(10mL)中，冰浴条件下，缓慢加入二乙胺基三氟化硫(771mg,4.79mmol,1.5eq)，随后恢复至室温条件下反应6小时。待反应完全后，加入饱和碳酸氢钠溶液淬灭，加入乙酸乙酯萃取，浓缩有机相，经正相柱层析纯化(石油醚:乙酸乙酯＝90:10)得黄色油状物8-[4-(二氟甲基)-3-甲氧基苯基]-1,4-二氧杂螺[4.5]癸-7-烯(752mg,收率69.6％)。LCMS(ESI)[M+H]+:297。1H NMR(400MHz,DMSO)δ7.43(d,J＝8.0Hz,1H),7.14–7.06(m,2H),7.19–6.88(m,1H),6.14(td,J＝3.9,2.0Hz,1H),3.92(s,4H),3.87(s,3H),2.58(m,J＝6.4,2.1Hz,2H),2.39(m,J＝2.1Hz,2H),1.82(t,J＝6.5Hz,2H)。2-Methoxy-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzaldehyde (1.0 g, 3.19 mmol, 1.0 eq) was dissolved in ultra-dry dichloromethane (10 mL). Diethylaminosulfur trifluoride (771 mg, 4.79 mmol, 1.5 eq) was slowly added in an ice bath. The mixture was then allowed to react at room temperature for 6 hours. After completion, the reaction was quenched with saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was concentrated and purified by normal phase column chromatography (petroleum ether:ethyl acetate = 90:10) to afford 8-[4-(difluoromethyl)-3-methoxyphenyl]-1,4-dioxaspiro[4.5]dec-7-ene (752 mg, 69.6% yield) as a yellow oil. LCMS (ESI) [M+H] + : 297. 1H NMR (400MHz, DMSO) δ7.43(d,J＝8.0Hz,1H),7.14–7.06(m,2H),7.19–6.88(m,1H),6.14(td,J＝3.9,2.0Hz ,1H),3.92(s,4H),3.87(s,3H),2.58(m,J＝6.4,2.1Hz,2H),2.39(m,J＝2.1Hz,2H),1.82(t,J＝6.5Hz,2H).

步骤3：8-[4-(二氟甲基)-3-甲氧基苯基]-1,4-二氧杂螺[4.5]癸烷的合成Step 3: Synthesis of 8-[4-(difluoromethyl)-3-methoxyphenyl]-1,4-dioxaspiro[4.5]decane

将8-[4-(二氟甲基)-3-甲氧基苯基]-1,4-二氧杂螺[4.5]癸-7-烯(700mg,2.37mmol,1.0eq)溶于无水甲醇(10mL)中，加入钯碳(70mg,10％)，在氢气氛下常温常压下反应2小时。待反应完全后，将反应液过滤，滤液浓缩得到黄色油状物8-[4-(二氟甲基)-3-甲氧基苯基]-1,4-二氧杂螺[4.5]癸烷(698mg,收率99.0％)。LCMS(ESI)[M+H]+:299。1H NMR(400MHz,DMSO)δ7.42(d,J＝7.9Hz,1H),7.18–6.87(m,1H),6.99(s,1H),6.94(dd,J＝7.8,1.4Hz,1H),3.91(d,J＝2.6Hz,4H),3.86(s,3H),2.66(m,J＝11.4,3.4Hz,1H),1.85–1.56(m,8H)。8-[4-(Difluoromethyl)-3-methoxyphenyl]-1,4-dioxaspiro[4.5]dec-7-ene (700 mg, 2.37 mmol, 1.0 eq) was dissolved in anhydrous methanol (10 mL), and palladium on carbon (70 mg, 10%) was added. The mixture was reacted under a hydrogen atmosphere at room temperature and pressure for 2 hours. After the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated to obtain 8-[4-(difluoromethyl)-3-methoxyphenyl]-1,4-dioxaspiro[4.5]decane (698 mg, 99.0% yield) as a yellow oil. LCMS (ESI) [M+H] + : 299. 1H NMR(400MHz,DMSO)δ7.42(d,J＝7.9Hz,1H),7.18–6.87(m,1H),6.99(s,1H),6.94(dd,J＝7.8,1 .4Hz,1H),3.91(d,J＝2.6Hz,4H),3.86(s,3H),2.66(m,J＝11.4,3.4Hz,1H),1.85–1.56(m,8H).

步骤4：4-[4-(二氟甲基)-3-甲氧基苯基]环己-1-酮的合成Step 4: Synthesis of 4-[4-(difluoromethyl)-3-methoxyphenyl]cyclohexan-1-one

将8-[4-(二氟甲基)-3-甲氧基苯基]-1,4-二氧杂螺[4.5]癸烷(650mg,2.18mmol,1.0eq)溶于丙酮(5mL)与水(5mL)中，加入对甲苯磺酸一水合物(124mg,0.65mmol,0.3eq)，60℃反应16小时。待反应完全后，加入乙酸乙酯萃取，浓缩有机相得黄色油状物4-[4-(二氟甲基)-3-甲氧基苯基]环己-1-酮(463mg,收率83.5％)。LCMS(ESI)[M+H]+:255。1H NMR(400MHz,DMSO)δ7.44(d,J＝7.9Hz,1H),7.11(d,J＝1.6Hz,1H),7.19–6.89(m,1H),7.01(dd,J＝7.8,1.5Hz,1H),3.87(s,3H),3.13(m,J＝12.0,3.5Hz,1H),2.60(m,J＝14.1,6.1Hz,2H),2.29(m,J＝14.5,4.3,2.1Hz,2H),2.10(m,J＝12.2,5.9,3.0Hz,2H),2.04–1.88(m,2H)。8-[4-(Difluoromethyl)-3-methoxyphenyl]-1,4-dioxaspiro[4.5]decane (650 mg, 2.18 mmol, 1.0 eq) was dissolved in acetone (5 mL) and water (5 mL). p-Toluenesulfonic acid monohydrate (124 mg, 0.65 mmol, 0.3 eq) was added and reacted at 60°C for 16 hours. After the reaction was complete, ethyl acetate was added for extraction. The organic phase was concentrated to obtain 4-[4-(difluoromethyl)-3-methoxyphenyl]cyclohexan-1-one (463 mg, 83.5% yield) as a yellow oil. LCMS (ESI) [M+H] + : 255. 1H NMR (400MHz, DMSO) δ7.44(d,J＝7.9Hz,1H),7.11(d,J＝1.6Hz,1H),7.19–6.89(m,1H),7.01(dd,J＝7.8,1.5Hz,1H),3.87(s,3H),3.13(m, J＝12.0,3.5Hz,1H),2.60(m,J＝14.1,6.1Hz,2H),2.29(m,J＝14.5,4.3,2.1Hz,2H),2.10(m,J＝12.2,5.9,3.0Hz,2H),2.04–1.88(m,2H).

步骤5：6-[4-(二氟甲基)-3-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 5: Synthesis of 6-[4-(difluoromethyl)-3-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-[4-(二氟甲基)-3-甲氧基苯基]环己-1-酮(400mg,1.57mmol,1.0eq)溶于超干四氢呋喃(6mL)中，分批加苯基三甲基三溴化铵(651mg,1.73mmol,1.1eq)，在常温常压下反应1小时。待反应完全后，用乙酸乙酯萃取，浓缩有机相，并用乙腈(4mL)重新溶解，随后加入硫脲(120mg,1.57mmol,1.0eq)，60℃反应4小时。待反应完全后，静置冷却至室温，过滤，得到类白色固体6-[4-(二氟甲基)-3-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(185mg)，未经进一步纯化，直接用于下一步反应。LCMS(ESI)[M+H]+:311。4-[4-(Difluoromethyl)-3-methoxyphenyl]cyclohexan-1-one (400 mg, 1.57 mmol, 1.0 eq) was dissolved in ultra-dry tetrahydrofuran (6 mL). Phenyltrimethylammonium tribromide (651 mg, 1.73 mmol, 1.1 eq) was added portionwise and allowed to react at room temperature and pressure for 1 hour. After the reaction was complete, the mixture was extracted with ethyl acetate, the organic phase was concentrated, and redissolved in acetonitrile (4 mL). Thiourea (120 mg, 1.57 mmol, 1.0 eq) was then added and allowed to react at 60°C for 4 hours. After the reaction was complete, the mixture was allowed to cool to room temperature and filtered to obtain 6-[4-(difluoromethyl)-3-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (185 mg) as an off-white solid, which was used directly in the next step without further purification. LCMS (ESI) [M+H] + : 311.

步骤6：2'-氯-N-{6-[4-(二氟甲基)-3-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of 2'-chloro-N-{6-[4-(difluoromethyl)-3-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将6-[4-(二氟甲基)-3-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(150mg,0.48mmol,1.0eq)溶于乙腈(3mL)中，依次加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(135mg,0.48mmol,1.0eq)和N-甲基咪唑(119mg,1.46mmol,3.0eq)，在常温常压下反应12小时。待反应完全后，制备纯化得白固体2'-氯-N-{6-[4-(二氟甲基)-3-甲氧基苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(8.0mg,收率3.04％)。LCMS(ESI)[M+H]+:571.14。1H NMR(400MHz,DMSO)δ8.78(s,1H),8.17(s,1H),7.52(s,1H),7.45–7.39(m,2H),7.14(s,1H),7.04(d,J＝1.7Hz,1H),7.19–6.87(m,1H),3.85(s,3H),3.62(s,3H),3.08(s,1H),2.98–2.78(m,2H),2.74(s,2H),2.58(s,3H),2.05(t,J＝6.6Hz,2H)。6-[4-(Difluoromethyl)-3-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (150 mg, 0.48 mmol, 1.0 eq) was dissolved in acetonitrile (3 mL). N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (135 mg, 0.48 mmol, 1.0 eq) and N-methylimidazole (119 mg, 1.46 mmol, 3.0 eq) were added sequentially. The mixture was allowed to react at room temperature and pressure for 12 hours. After the reaction was complete, 2'-chloro-N-{6-[4-(difluoromethyl)-3-methoxyphenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (8.0 mg, 3.04% yield) was obtained as a white solid. LCMS(ESI)[M+H] + :571.14. 1H NMR (400MHz, DMSO) δ8.78(s,1H),8.17(s,1H),7.52(s,1H),7.45–7.39(m,2H),7.14(s,1H),7.04(d,J＝1.7Hz,1H),7.19– 6.87(m,1H),3.85(s,3H),3.62(s,3H),3.08(s,1H),2.98–2.78(m,2H),2.74(s,2H),2.58(s,3H),2.05(t,J＝6.6Hz,2H).

实施例30：2'-氯-5'-甲氧基-6-甲基-N-{6-[2-(2,2,2-三氟乙基)氨基]嘧啶-5-基}-4,5,6,7-四氢苯并[d]噻唑-2-基-[4,4'-联吡啶]-3-甲酰胺(化合物52)的合成
Example 30: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[2-(2,2,2-trifluoroethyl)amino]pyrimidin-5-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide (Compound 52)

步骤1：5-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-N-(2,2,2-三氟乙基)嘧啶-2-胺的合成Step 1: Synthesis of 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine

将2-氯-5-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)嘧啶(5.00g,20.8mmol,1.0eq)、2,2,2-三氟乙胺(8.43g,62.5mmol,3.0eq)和二异丙基乙胺(26.8g,208.3mmol,10.0eq)加入到N,N-二甲基甲酰胺(30mL)中，80℃反应24小时。反应结束后，将反应液浓缩，加入乙酸乙酯和水萃取分液，有机相浓缩后得到黄色固体粗品5-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-N-(2,2,2-三氟乙基)嘧啶-2-胺(8.0g)。LCMS(TOF MS ES+)m/z[M+H]+:304.13。2-Chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (5.00 g, 20.8 mmol, 1.0 eq), 2,2,2-trifluoroethylamine (8.43 g, 62.5 mmol, 3.0 eq), and diisopropylethylamine (26.8 g, 208.3 mmol, 10.0 eq) were added to N,N-dimethylformamide (30 mL) and reacted at 80°C for 24 hours. After completion of the reaction, the reaction solution was concentrated and extracted with ethyl acetate and water. The organic phase was concentrated to obtain a crude yellow solid, 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine (8.0 g). LCMS (TOF MS ES+)m/z[M+H] + :304.13.

步骤2：5-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)-N-(2,2,2-三氟乙基)嘧啶-2-胺的合成Step 2: Synthesis of 5-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine

将5-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-N-(2,2,2-三氟乙基)嘧啶-2-胺(3.0g,9.9mmol,1.0eq)、1,4-二氧阿司匹林[4.5]癸-7-烯-8-基三氟甲磺酸酯(2.85g,9.9mmol,1.0eq)、1,1-双(二苯基膦)二荗铁二氯化钯(723mg,0.99mmol,0.1eq)和碳酸钾(3.41g,24.7mmol,2.5eq)加入到1,4-二氧六环(40mL)和水(8mL)中，80℃反应2小时。反应结束后，将反应液过滤，滤液加入乙酸乙酯和水萃取，正相柱层析纯化得到白色固体5-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)-N-(2,2,2-三氟乙基)嘧啶-2-胺(1.2g,收率38.3％)。LCMS(TOF MS ES+)m/z[M+H]+:316.11。1H NMR(400MHz,DMSO)δ8.46(s,2H),7.78(t,J＝6.7Hz,1H),5.99(m,J＝3.9,1.9Hz,1H),4.15(m,J＝9.7,6.7Hz,2H),3.93(s,4H),2.50(d,J＝7.5Hz,2H),2.35(dd,J＝4.1,2.2Hz,2H),1.81(t,J＝6.5Hz,2H)。5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine (3.0 g, 9.9 mmol, 1.0 eq), 1,4-dioxaspirino[4.5]dec-7-en-8-yl trifluoromethanesulfonate (2.85 g, 9.9 mmol, 1.0 eq), 1,1-bis(diphenylphosphino)dibenzoylmethanepalladium chloride (723 mg, 0.99 mmol, 0.1 eq) and potassium carbonate (3.41 g, 24.7 mmol, 2.5 eq) were added to 1,4-dioxane (40 mL) and water (8 mL) and reacted at 80 ° C for 2 hours. After the reaction, the reaction mixture was filtered, and the filtrate was extracted with ethyl acetate and water. The product was purified by normal phase column chromatography to obtain 5-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine (1.2 g, 38.3% yield) as a white solid. LCMS (TOF MS ES+) m/z [M+H] + : 316.11. 1H NMR (400MHz, DMSO) δ8.46 (s, 2H), 7.78 (t, J = 6.7Hz, 1H), 5.99 (m, J = 3.9, 1.9Hz, 1H), 4.15 (m, J = 9.7, 6.7Hz, 2H), 3.93 (s, 4H), 2.50 (d, J = 7.5Hz, 2H), 2.35 (dd, J = 4.1, 2.2Hz, 2H), 1.81 (t, J = 6.5Hz, 2H).

步骤3：5-(1,4-二氧阿司匹林[4.5]癸-8-基)-N-(2,2,2-三氟乙基)嘧啶-2-胺的合成Step 3: Synthesis of 5-(1,4-dioxyaspirino[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine

将5-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)-N-(2,2,2-三氟乙基)嘧啶-2-胺(1.2g,3.8mmol,1.0eq)和钯碳(240mg)加入到四氢呋喃(30mL)中，室温下搅拌1小时。待反应结束，采用硅藻土过滤，滤液浓缩后得到白色固体5-(1,4-二氧阿司匹林[4.5]癸-8-基)-N-(2,2,2-三氟乙基)嘧啶-2-胺(1.1g,收率91.0％)，直接用于下一步反应。LCMS(TOF MS ES+)m/z[M+H]+:318.13。1H NMR(400MHz,DMSO)δ8.26(s,2H),7.58(t,J＝6.7Hz,1H),4.11(m,J＝9.6,6.7Hz,2H),3.89(d,J＝2.4Hz,4H),2.48(d,J＝10.8Hz,1H),1.77(d,J＝9.7Hz,4H),1.71–1.54(m,4H)。5-(1,4-Dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine (1.2 g, 3.8 mmol, 1.0 eq) and palladium on carbon (240 mg) were added to tetrahydrofuran (30 mL) and stirred at room temperature for 1 hour. After the reaction was complete, the mixture was filtered through celite and the filtrate was concentrated to afford 5-(1,4-Dioxyaspirino[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine (1.1 g, 91.0% yield) as a white solid, which was used directly in the next reaction. LCMS (TOF MS ES+) m/z [M+H] + : 318.13. 1H NMR (400MHz, DMSO) δ8.26 (s, 2H), 7.58 (t, J = 6.7Hz, 1H), 4.11 (m, J = 9.6, 6.7Hz, 2H), 3. 89(d,J＝2.4Hz,4H),2.48(d,J＝10.8Hz,1H),1.77(d,J＝9.7Hz,4H),1.71–1.54(m,4H).

步骤4：4-[2-(2,2,2-三氟乙基)氨基]嘧啶-5-基环己烷-1-酮的合成Step 4: Synthesis of 4-[2-(2,2,2-trifluoroethyl)amino]pyrimidin-5-ylcyclohexane-1-one

将5-(1,4-二氧阿司匹林[4.5]癸-8-基)-N-(2,2,2-三氟乙基)嘧啶-2-胺(200mg,0.63mmol,1.0eq)和对甲苯磺酸一水合物(180mg,0.94mmol,1.5eq)加入到丙酮(6mL)和水(2mL)中，60℃反应1小时。反应结束后，加入乙酸乙酯和水萃取，正相柱分离纯化得到黄色固体4-[2-(2,2,2-三氟乙基)氨基]嘧啶-5-基环己烷-1-酮(171mg,收率99.0％)。LCMS(TOF MS ES+)m/z[M+H]+:274.10。1H NMR(400MHz,DMSO)δ8.35(s,2H),7.63(t,J＝6.7Hz,1H),4.12(m,J＝9.6,6.6Hz,2H),2.97(m,J＝12.2,3.5Hz,1H),2.58(m,J＝14.9,7.5Hz,2H),2.27(d,J＝14.4Hz,2H),2.12–2.01(m,2H),1.89(m,J＝13.1,4.1Hz,2H)。5-(1,4-Dioxyaspirino[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)pyrimidin-2-amine (200 mg, 0.63 mmol, 1.0 eq) and p-toluenesulfonic acid monohydrate (180 mg, 0.94 mmol, 1.5 eq) were added to acetone (6 mL) and water (2 mL) and reacted at 60°C for 1 hour. After completion of the reaction, the mixture was extracted with ethyl acetate and water. The product was separated and purified using a normal phase column to obtain a yellow solid, 4-[2-(2,2,2-trifluoroethyl)amino]pyrimidin-5-ylcyclohexane-1-one (171 mg, 99.0% yield). LCMS (TOF MS ES+) m/z [M+H] + : 274.10. 1H NMR (400MHz, DMSO) δ8.35(s,2H),7.63(t,J＝6.7Hz,1H),4.12(m,J＝9.6,6.6Hz,2H),2.97(m,J＝12.2,3.5Hz ,1H),2.58(m,J＝14.9,7.5Hz,2H),2.27(d,J＝14.4Hz,2H),2.12–2.01(m,2H),1.89(m,J＝13.1,4.1Hz,2H).

步骤5：6-{2-[(2,2,2-三氟乙基)氨基]嘧啶-5-基}-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 5: Synthesis of 6-{2-[(2,2,2-trifluoroethyl)amino]pyrimidin-5-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-[2-(2,2,2-三氟乙基)氨基]嘧啶-5-基环己烷-1-酮(150mg,0.55mmol,1.0eq)、单质碘(167mg,0.66mmol,1.2eq)和硫脲(83mg,1.10mmol,2.0eq)加入到乙醇(5mL)中，100℃下搅拌2小时。反应结束后，加入碳酸氢钠溶液和硫代硫酸钠溶液，再加入乙酸乙酯和水萃取，正相柱分离纯化，得到黄色固体6-{2-[(2,2,2-三氟乙基)氨基]嘧啶-5-基}-4,5,6,7-四氢苯并[d]噻唑-2-胺(172mg,收率94.9％)。LCMS(TOF MS ES+)m/z[M+H]+:330.09。4-[2-(2,2,2-trifluoroethyl)amino]pyrimidin-5-ylcyclohexane-1-one (150 mg, 0.55 mmol, 1.0 eq), iodine (167 mg, 0.66 mmol, 1.2 eq), and thiourea (83 mg, 1.10 mmol, 2.0 eq) were added to ethanol (5 mL) and stirred at 100°C for 2 hours. After completion of the reaction, sodium bicarbonate solution and sodium thiosulfate solution were added, followed by extraction with ethyl acetate and water. Purification by normal phase column chromatography afforded 6-{2-[(2,2,2-trifluoroethyl)amino]pyrimidin-5-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (172 mg, 94.9% yield) as a yellow solid. LCMS (TOF MS ES+) m/z [M+H] + : 330.09.

步骤6：2'-氯-5'-甲氧基-6-甲基-N-{6-[2-(2,2,2-三氟乙基)氨基]嘧啶-5-基}-4,5,6,7-四氢苯并[d]噻唑-2-基-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[2-(2,2,2-trifluoroethyl)amino]pyrimidin-5-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide

将6-{2-[(2,2,2-三氟乙基)氨基]嘧啶-5-基}-4,5,6,7-四氢苯并[d]噻唑-2-胺(150mg,0.45mmol,1.0eq)、2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(126mg,0.45mmol,1.0eq)、N,N,N',N'-四甲基氯甲脒六氟磷酸盐(191mg,0.68mmol,1.5eq)和N-甲基咪唑(149mg,1.82mmol,4.0eq)加入到乙腈(6mL)中，室温反应1小时。反应结束后，浓缩反应液，采用高效液相制备色谱分纯化得到白色固体2'-氯-5'-甲氧基-6-甲基-N-{6-[2-(2,2,2-三氟乙基)氨基]嘧啶-5-基}-4,5,6,7-四氢苯并[d]噻唑-2-基-[4,4'-联吡啶]-3-甲酰胺(62.3mg,收率23.2％)。LCMS(TOF MS ES+)m/z[M+H]+:590.1340。1H NMR(400MHz,DMSO)δ12.49(s,1H),8.77(s,1H),8.35(s,2H),8.17(s,1H),7.63(t,J＝6.7Hz,1H),7.51(s,1H),7.41(s,1H),4.11(dd,J＝10.1,7.2Hz,2H),3.62(s,3H),2.92(d,J＝14.3Hz,2H),2.73(q,J＝15.0Hz,3H),2.58(s,3H),2.00(d,J＝7.3Hz,2H)。6-{2-[(2,2,2-trifluoroethyl)amino]pyrimidin-5-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (150 mg, 0.45 mmol, 1.0 eq), 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (126 mg, 0.45 mmol, 1.0 eq), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (191 mg, 0.68 mmol, 1.5 eq) and N-methylimidazole (149 mg, 1.82 mmol, 4.0 eq) were added to acetonitrile (6 mL) and reacted at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated and purified by preparative HPLC to yield 2'-chloro-5'-methoxy-6-methyl-N-{6-[2-(2,2,2-trifluoroethyl)amino]pyrimidin-5-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide (62.3 mg, 23.2% yield) as a white solid. LCMS (TOF MS ES+) m/z [M+H] + : 590.1340. 1H NMR (400MHz, DMSO) δ12.49(s,1H),8.77(s,1H),8.35(s,2H),8.17(s,1H),7.63(t,J＝6.7Hz,1H),7.51(s,1H),7.41(s,1H),4 .11(dd,J＝10.1,7.2Hz,2H),3.62(s,3H),2.92(d,J＝14.3Hz,2H),2.73(q,J＝15.0Hz,3H),2.58(s,3H),2.00(d,J＝7.3Hz,2H).

实施例31：2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-基-[4,4'-联吡啶]-3-甲酰胺(化合物54)的合成
Example 31: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide (Compound 54)

步骤1：4-溴-N-(2,2,2-三氟乙基)苯胺的合成Step 1: Synthesis of 4-bromo-N-(2,2,2-trifluoroethyl)aniline

将(4-溴苯基)硼酸(5.00g,24.9mmol,1.0eq)、2,2,2-三氟乙胺(7.39g,74.7mmol,3.0eq)、醋酸铜(6.76g,37.3mmol,1.5eq)和三乙胺(7.54g,74.7mmol,3.0eq)加入到乙腈(100mL)中，室温反应16小时，然后80℃反应1小时。反应结束后，硅藻土过滤除去不溶物，浓缩滤液，正相柱层析纯化得到无色油状物4-溴-N-(2,2,2-三氟乙基)苯胺(4.15g,收率65.4％)。LCMS(TOF MS ES+)m/z[M+H]+:253.97。1H NMR(400MHz,DMSO)δ7.30–7.20(m,2H),6.76–6.66(m,2H),6.44(t,J＝7.0Hz,1H),3.93(m,J＝9.7,6.9Hz,2H)。(4-Bromophenyl)boronic acid (5.00 g, 24.9 mmol, 1.0 eq), 2,2,2-trifluoroethylamine (7.39 g, 74.7 mmol, 3.0 eq), copper acetate (6.76 g, 37.3 mmol, 1.5 eq), and triethylamine (7.54 g, 74.7 mmol, 3.0 eq) were added to acetonitrile (100 mL) and reacted at room temperature for 16 hours, followed by 80°C for 1 hour. After completion of the reaction, the insoluble material was removed by filtration through Celite, and the filtrate was concentrated and purified by normal phase column chromatography to afford 4-bromo-N-(2,2,2-trifluoroethyl)aniline (4.15 g, 65.4% yield) as a colorless oil. LCMS (TOF MS ES+) m/z [M+H] + : 253.97. 1 H NMR (400MHz, DMSO) δ7.30–7.20 (m, 2H), 6.76–6.66 (m, 2H), 6.44 (t, J = 7.0Hz, 1H), 3.93 (m, J = 9.7, 6.9Hz, 2H).

步骤2：4-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)-N-(2,2,2-三氟乙基)苯胺的合成Step 2: Synthesis of 4-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)aniline

将4-溴-N-(2,2,2-三氟乙基)苯胺(4.15g,16.3mmol,1.0eq)、4,4,5,5-四甲基-2-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)-1,3,2-二氧杂硼烷(5.21g,19.6mmol,1.5eq)、1,1-双(二苯基膦)二荗铁二氯化钯(1.19g,1.63mmol,0.1eq)和碳酸钾(5.64g,40.8mmol,2.5eq)加入到1,4-二氧六环(60mL)和水(12mL)中，90℃反应24小时。反应结束后，过滤，滤液加入乙酸乙酯和水萃取，正相柱层析纯化得到黄色固体4-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)-N-(2,2,2-三氟乙基)苯胺(2.1g,收率41.1％)。LCMS(TOF MS ES+)m/z[M+H]+:314.12。1H NMR(400MHz,DMSO)δ7.24–7.16(m,2H),6.73–6.65(m,2H),6.23(m,J＝6.8Hz,1H),5.86–5.79(m,1H),3.97–3.88(m,6H),2.50(d,J＝10.3Hz,2H),2.36–2.32(m,2H),1.79(t,J＝6.5Hz,2H)。4-Bromo-N-(2,2,2-trifluoroethyl)aniline (4.15 g, 16.3 mmol, 1.0 eq), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (5.21 g, 19.6 mmol, 1.5 eq), 1,1-bis(diphenylphosphino)diborane iron palladium dichloride (1.19 g, 1.63 mmol, 0.1 eq) and potassium carbonate (5.64 g, 40.8 mmol, 2.5 eq) were added to 1,4-dioxane (60 mL) and water (12 mL) and reacted at 90 ° C for 24 hours. After the reaction, the reaction mixture was filtered, and the filtrate was extracted with ethyl acetate and water. The mixture was then purified by normal phase column chromatography to obtain a yellow solid of 4-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)aniline (2.1 g, 41.1% yield). LCMS (TOF MS ES+) m/z [M+H] + : 314.12. 1H NMR (400MHz, DMSO) δ7.24–7.16(m,2H),6.73–6.65(m,2H),6.23(m,J＝6.8Hz,1H),5.86–5.79 (m,1H),3.97–3.88(m,6H),2.50(d,J＝10.3Hz,2H),2.36–2.32(m,2H),1.79(t,J＝6.5Hz,2H).

步骤3：4-(1,4-二氧阿司匹林[4.5]癸-8-基)-N-(2,2,2-三氟乙基)苯胺的合成Step 3: Synthesis of 4-(1,4-dioxyaspirino[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)aniline

将4-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)-N-(2,2,2-三氟乙基)苯胺(2.1g,6.7mmol,1.0eq)和钯碳(210mg)加入到四氢呋喃(40mL)中，室温下搅拌1小时。反应结束后，硅藻土过滤，滤液浓缩后得到黄色固体4-(1,4-二氧阿司匹林[4.5]癸-8-基)-N-(2,2,2-三氟乙基)苯胺(2.1g，收率99.4％)，直接用于下一步反应。LCMS(TOF MS ES+)m/z[M+H]+:316.14。4-(1,4-Dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)aniline (2.1 g, 6.7 mmol, 1.0 eq) and palladium on carbon (210 mg) were added to tetrahydrofuran (40 mL) and stirred at room temperature for 1 hour. After the reaction, the mixture was filtered through celite and the filtrate was concentrated to afford 4-(1,4-Dioxyaspirino[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)aniline (2.1 g, 99.4% yield) as a yellow solid, which was used directly in the next reaction. LCMS (TOF MS ES+) m/z [M+H] + : 316.14.

步骤4：4-[4-(2,2,2-三氟乙基)氨基]苯基环己烷-1-酮的合成Step 4: Synthesis of 4-[4-(2,2,2-trifluoroethyl)amino]phenylcyclohexane-1-one

将4-(1,4-二氧阿司匹林[4.5]癸-8-基)-N-(2,2,2-三氟乙基)苯胺(2.1g,6.66mmol,1.0eq)和对甲苯磺酸一水合物(2.53g,13.33mmol,2.0eq)加入到丙酮(30mL)和水(10mL)中，60℃反应1小时。反应结束后，加入乙酸乙酯和水萃取，正相柱分离纯化得到紫色固体4-[4-(2,2,2-三氟乙基)氨基]苯基环己烷-1-酮(3.6g,收率85.0％)。LCMS(TOF MS ES+)m/z[M+H]+:272.11。1H NMR(400MHz,DMSO)δ7.07–6.98(m,2H),6.70–6.62(m,2H),6.05(t,J＝7.0Hz,1H),3.86(m,J＝9.8,6.9Hz,2H),2.90(m,J＝12.0,3.4Hz,1H),2.55(m,J＝14.1,7.1Hz,2H),2.23(m,J＝14.7,4.5,2.2Hz,2H),2.01(m,J＝12.6,3.0Hz,2H),1.79(m,J＝13.2,4.1Hz,2H)。4-(1,4-Dioxyaspirino[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)aniline (2.1 g, 6.66 mmol, 1.0 eq) and p-toluenesulfonic acid monohydrate (2.53 g, 13.33 mmol, 2.0 eq) were added to acetone (30 mL) and water (10 mL) and reacted at 60°C for 1 hour. After completion of the reaction, the mixture was extracted with ethyl acetate and water. The product was separated and purified by normal phase column chromatography to obtain a purple solid, 4-[4-(2,2,2-trifluoroethyl)amino]phenylcyclohexane-1-one (3.6 g, 85.0% yield). LCMS (TOF MS ES+) m/z [M+H] + : 272.11. 1H NMR (400MHz, DMSO) δ7.07–6.98(m,2H),6.70–6.62(m,2H),6.05(t,J＝7.0Hz,1H),3.86(m,J＝9.8,6.9Hz,2H),2.90(m,J＝12.0,3. 4Hz, 1H), 2.55 (m, J = 14.1, 7.1Hz, 2H), 2.23 (m, J = 14.7, 4.5, 2.2Hz, 2H), 2.01 (m, J = 12.6, 3.0Hz, 2H), 1.79 (m, J = 13.2, 4.1Hz, 2H).

步骤5：6-{4-[(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 5: Synthesis of 6-{4-[(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-[4-(2,2,2-三氟乙基)氨基]苯基环己烷-1-酮(200mg,0.74mmol,1.0eq)、单质碘(206mg,0.81mmol,1.1eq)和硫脲(112mg,1.48mmol,2.0eq)加入到乙醇(40mL)中，80℃下搅拌16小时。反应结束后，加入碳酸氢钠溶液和硫代硫酸钠溶液，再加入乙酸乙酯和水萃取，正相柱分离纯化得到黄色固体6-{4-[(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-胺(210mg,收率87.0％)。LCMS(TOF MS ES+)m/z[M+H]+:328.10。4-[4-(2,2,2-trifluoroethyl)amino]phenylcyclohexane-1-one (200 mg, 0.74 mmol, 1.0 eq), iodine (206 mg, 0.81 mmol, 1.1 eq), and thiourea (112 mg, 1.48 mmol, 2.0 eq) were added to ethanol (40 mL) and stirred at 80°C for 16 hours. After completion of the reaction, sodium bicarbonate solution and sodium thiosulfate solution were added, followed by extraction with ethyl acetate and water. The product was separated and purified by normal phase column chromatography to obtain 6-{4-[(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (210 mg, 87.0% yield) as a yellow solid. LCMS (TOF MS ES+) m/z [M+H] + : 328.10.

步骤6：2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-基-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide

将6-{4-[(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-胺(200mg,0.61mmol,1.0eq)、2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(170mg,0.61mmol,1.0eq)、N,N,N',N'-四甲基氯甲脒六氟磷酸盐(257mg,0.91mmol,1.5eq)和N-甲基咪唑(200mg,2.44mmol,4.0eq)加入到乙腈(10mL)中，室温反应1小时。反应结束后，将反应液浓缩，经高效液相制备色谱分离得到白色固体2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-基-[4,4'-联吡啶]-3-甲酰胺(35.7mg,收率9.9％)。LCMS(TOF MS ES+)m/z[M+H]+:588.1451。1H NMR(400MHz,DMSO)δ12.45(s,1H),8.77(s,1H),8.17(s,1H),7.51(s,1H),7.41(s,1H),7.07(d,J＝8.2Hz,2H),6.67(d,J＝8.2Hz,2H),6.07(t,J＝7.0Hz,1H),3.88(m,J＝9.9,7.0Hz,2H),3.62(s,3H),2.86(d,J＝13.0Hz,2H),2.69(d,J＝8.7Hz,3H),2.58(s,3H),2.02–1.85(m,2H)。6-{4-[(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (200 mg, 0.61 mmol, 1.0 eq), 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (170 mg, 0.61 mmol, 1.0 eq), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (257 mg, 0.91 mmol, 1.5 eq) and N-methylimidazole (200 mg, 2.44 mmol, 4.0 eq) were added to acetonitrile (10 mL) and reacted at room temperature for 1 hour. After completion of the reaction, the reaction solution was concentrated and separated by preparative HPLC to afford 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide (35.7 mg, 9.9% yield) as a white solid. LCMS (TOF MS ES+) m/z [M+H] + : 588.1451. 1H NMR (400MHz, DMSO) δ12.45(s,1H),8.77(s,1H),8.17(s,1H),7.51(s,1H),7.41(s,1H),7.07(d,J＝8.2Hz,2H),6.67(d,J＝8.2Hz,2H),6.07 (t,J＝7.0Hz,1H),3.88(m,J＝9.9,7.0Hz,2H),3.62(s,3H),2.86(d,J＝13.0Hz,2H),2.69(d,J＝8.7Hz,3H),2.58(s,3H),2.02–1.85(m,2H).

实施例32：2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(N-甲基氨基磺酰基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(化合物49)的合成
Example 32: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(N-methylaminosulfonyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (Compound 49)

步骤1：N-甲基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯磺酰胺的合成Step 1: Synthesis of N-methyl-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzenesulfonamide

将4-溴-N-甲基苯磺酰胺(8g,32mmol,1.0eq)、4,4,5,5-四甲基-2-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)-1,3,2-二氧硼烷(8.5g,32mmol,1.0eq)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(2.3g,3.2mmol,0.1eq)和碳酸钾(8.8g,64mmol,2.0eq)溶于1,4-二氧六环(20mL)和水(4mL)中，氮气保护下，80℃下反应3小时。反应完全后，将反应液过滤，滤液浓缩、拌样，然后正相柱分离纯化(乙酸乙酯/石油醚，乙酸乙酯％＝20％)，得到黄色固体N-甲基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯磺酰胺(4.7g,收率47.9％)。LCMS(ESI)[M+H]:310.1099。4-Bromo-N-methylbenzenesulfonamide (8 g, 32 mmol, 1.0 eq), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (8.5 g, 32 mmol, 1.0 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (2.3 g, 3.2 mmol, 0.1 eq) and potassium carbonate (8.8 g, 64 mmol, 2.0 eq) were dissolved in 1,4-dioxane (20 mL) and water (4 mL) and reacted at 80 ° C for 3 hours under nitrogen protection. After the reaction was complete, the reaction solution was filtered, the filtrate was concentrated, the sample was stirred, and then purified by normal phase column separation (ethyl acetate/petroleum ether, ethyl acetate% = 20%) to obtain N-methyl-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzenesulfonamide (4.7 g, yield 47.9%) as a yellow solid. LCMS (ESI) [M+H]: 310.1099.

步骤2：N-甲基-4-(1,4-二氧杂螺[4.5]癸烷-8-基)苯磺酰胺的合成Step 2: Synthesis of N-methyl-4-(1,4-dioxaspiro[4.5]decan-8-yl)benzenesulfonamide

将N-甲基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯磺酰胺(2.4g,6.4mmol,1.0eq)、钯碳(480mg)溶于甲醇(5mL)中，氢气置换三次，室温下反应16小时。反应完全后，将混合液过滤，滤液浓缩得到白色固体N-甲基-4-(1,4-二氧杂螺[4.5]癸烷-8-基)苯磺酰胺(2.4g)，直接用于下一步反应。LCMS(ESI)[M+H]:310.127。N-methyl-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzenesulfonamide (2.4 g, 6.4 mmol, 1.0 eq) and palladium on carbon (480 mg) were dissolved in methanol (5 mL). The mixture was replaced with hydrogen three times and allowed to react at room temperature for 16 hours. After the reaction was complete, the mixture was filtered and the filtrate was concentrated to obtain N-methyl-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzenesulfonamide (2.4 g) as a white solid, which was used directly in the next reaction. LCMS (ESI) [M+H]: 310.127.

步骤3：N-甲基-4-(4-氧代环己基)苯磺酰胺的合成Step 3: Synthesis of N-methyl-4-(4-oxocyclohexyl)benzenesulfonamide

N-甲基-4-(1,4-二氧杂螺[4.5]癸烷-8-基)苯磺酰胺(800g,2.57mmol,1.0eq)、对甲基苯磺酸水合物(977mg,5.1mmol,2.0eq)溶于丙酮(9mL)和水(3mL)中，60℃反应16小时。反应完全后，混合液用碳酸氢钠溶液淬灭，再用乙酸乙酯和水萃取，有机相用无水硫酸钠除去多余水分，浓缩得到白色固体N-甲基-4-(4-氧代环己基)苯磺酰胺(627mg,收率91％)。LCMS(ESI)[M+H]:310.127。N-Methyl-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzenesulfonamide (800 g, 2.57 mmol, 1.0 eq) and p-toluenesulfonic acid hydrate (977 mg, 5.1 mmol, 2.0 eq) were dissolved in acetone (9 mL) and water (3 mL) and reacted at 60°C for 16 hours. After the reaction was complete, the mixture was quenched with sodium bicarbonate solution and extracted with ethyl acetate and water. The organic phase was dehydrated with anhydrous sodium sulfate and concentrated to yield N-methyl-4-(4-oxocyclohexyl)benzenesulfonamide (627 mg, 91% yield) as a white solid. LCMS (ESI) [M+H]: 310.127.

步骤4：4-(2-氨基-4,5,6,7-四氢苯并噻唑-6-基)-N-甲基苯磺酰胺的合成Step 4: Synthesis of 4-(2-amino-4,5,6,7-tetrahydrobenzothiazol-6-yl)-N-methylbenzenesulfonamide

将N-甲基-4-(4-氧代环己基)苯磺酰胺(100mg,0.37mmol,1.0eq)、硫脲(57mg,0.75mmol,2.0eq)和碘单质(114mg,0.56mmol,1.2eq)溶于乙醇(2mL)中，80℃下反应2小时。反应完全后，将混合液浓缩，然后用饱和的碳酸氢钠溶液和硫代硫酸钠溶液淬灭，再用乙酸乙酯和水萃取，得到黄色油状粗品4-(2-氨基-4,5,6,7-四氢苯并噻唑-6-基)-N-甲基苯磺酰胺(121mg)。1H NMR(400MHz,DMSO)δ7.77–7.68(m,2H),7.60–7.52(m,2H),7.42(s,1H),3.19(m,J＝12.0,3.5Hz,1H),2.61(td,J＝14.2,6.0Hz,2H),2.42(s,3H),2.30(m,J＝14.7,2.1Hz,2H),2.11(m,J＝12.7,6.0,3.0Hz,2H),1.94(m,J＝13.1,6.1Hz,2H)。N-methyl-4-(4-oxocyclohexyl)benzenesulfonamide (100 mg, 0.37 mmol, 1.0 eq), thiourea (57 mg, 0.75 mmol, 2.0 eq), and iodine (114 mg, 0.56 mmol, 1.2 eq) were dissolved in ethanol (2 mL) and reacted at 80°C for 2 hours. After the reaction was complete, the mixture was concentrated and then quenched with saturated sodium bicarbonate solution and sodium thiosulfate solution. The mixture was then extracted with ethyl acetate and water to obtain crude 4-(2-amino-4,5,6,7-tetrahydrobenzothiazol-6-yl)-N-methylbenzenesulfonamide (121 mg) as a yellow oil. 1H NMR (400MHz, DMSO) δ7.77–7.68(m,2H),7.60–7.52(m,2H),7.42(s,1H),3.19(m,J＝12.0,3.5Hz,1H),2.61(td,J＝14.2 ,6.0Hz,2H),2.42(s,3H),2.30(m,J＝14.7,2.1Hz,2H),2.11(m,J＝12.7,6.0,3.0Hz,2H),1.94(m,J＝13.1,6.1Hz,2H).

步骤5：2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(N-甲基氨基磺酰基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(N-methylaminosulfonyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide

将4-(2-氨基-4,5,6,7-四氢苯并噻唑-6-基)-N-甲基苯磺酰胺(120mg,0.37mmol,1.0eq)、2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(103mg,0.37mmol,1.0eq)、N,N,N,N-四甲基氯甲眯六氟磷酸盐(312mg,1.11mmol,3.0eq)和N-甲基咪唑(610mg,3.7mmol,20.0eq)溶于乙腈(5mL)中，室温反应2小时。反应完全后，制备纯化得到黄色粉末2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(N-甲基氨基磺酰基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(13.5mg,收率6％)。1H NMR(400MHz,DMSO)δ12.50(s,1H),8.78(s,1H),8.17(s,1H),7.72(d,J＝7.9Hz,2H),7.57(d,J＝8.1Hz,2H),7.52(s,1H),7.43–7.34(m,2H),3.62(s,3H),3.15(s,1H),2.97(d,J＝17.7Hz,1H),2.83–2.66(m,3H),2.58(s,3H),2.41(d,J＝4.9Hz,3H),2.05(d,J＝7.6Hz,2H)。4-(2-Amino-4,5,6,7-tetrahydrobenzothiazol-6-yl)-N-methylbenzenesulfonamide (120 mg, 0.37 mmol, 1.0 eq), 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (103 mg, 0.37 mmol, 1.0 eq), N,N,N,N-tetramethylchloromethylimidazole hexafluorophosphate (312 mg, 1.11 mmol, 3.0 eq) and N-methylimidazole (610 mg, 3.7 mmol, 20.0 eq) were dissolved in acetonitrile (5 mL) and reacted at room temperature for 2 hours. After the reaction was completed, the reaction mixture was purified to give a yellow powder of 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(N-methylaminosulfonyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (13.5 mg, yield 6%). 1H NMR (400MHz, DMSO) δ12.50(s,1H),8.78(s,1H),8.17(s,1H),7.72(d,J＝7.9Hz,2H),7.57(d,J＝8.1Hz,2H),7.52(s,1H),7.43–7.34(m ,2H),3.62(s,3H),3.15(s,1H),2.97(d,J＝17.7Hz,1H),2.83–2.66(m,3H),2.58(s,3H),2.41(d,J＝4.9Hz,3H),2.05(d,J＝7.6Hz,2H).

实施例33：2'-氯-N-{6-[4-(N,N-二甲基氨磺酰基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物50)的合成
Example 33: Synthesis of 2'-chloro-N-{6-[4-(N,N-dimethylsulfamoyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 50)

步骤1：N,N-二甲基-4-{1,4-二氧杂螺[4.5]癸-8-基}苯磺酰胺的合成Step 1: Synthesis of N,N-dimethyl-4-{1,4-dioxaspiro[4.5]dec-8-yl}benzenesulfonamide

将N-甲基-4-(1,4-二氧杂螺[4.5]癸-8-基)苯磺酰胺(600mg,1.85mmol,1.0eq)溶于超干的N,N-二甲基甲酰胺溶剂(7.0mL)中，0℃条件下，缓慢加入氢化钠(153.6mg,3.86mmol,2.0eq)和碘甲烷(142mg,2.89mmol,1.5eq)，加完后移至室温反应3小时。反应完全后，氯化铵溶液淬灭，乙酸乙酯和水萃取，干燥浓缩有机相，正相柱分离得到黄色固体N,N-二甲基-4-{1,4-二氧杂螺[4.5]癸-8-基}苯磺酰胺(620mg,收率98.9％).LCMS(ESI)[M+H]:326.1416.1H NMR(400MHz,DMSO)δ7.66(d,J＝8.0Hz,2H),7.50(d,J＝8.0Hz,2H),3.91–3.87(m,4H),2.77–2.55(m,7H),1.84–1.52(m,8H)。Dissolve N-methyl-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzenesulfonamide (600 mg, 1.85 mmol, 1.0 eq) in ultra-dry N,N-dimethylformamide solvent (7.0 mL). Slowly add sodium hydride (153.6 mg, 3.86 mmol, 2.0 eq) and iodomethane (142 mg, 2.89 mmol, 1.5 eq) at 0°C. After addition, move to room temperature and react for 3 hours. After the reaction was complete, the product was quenched with ammonium chloride solution and extracted with ethyl acetate and water. The organic phase was dried and concentrated, and separated by normal phase column chromatography to give N,N-dimethyl-4-{1,4-dioxaspiro[4.5]dec-8-yl}benzenesulfonamide (620 mg, yield 98.9%) as a yellow solid. LCMS (ESI) [M+H]: 326.1416. 1 H NMR (400 MHz, DMSO) δ 7.66 (d, J = 8.0 Hz, 2H), 7.50 (d, J = 8.0 Hz, 2H), 3.91–3.87 (m, 4H), 2.77–2.55 (m, 7H), 1.84–1.52 (m, 8H).

步骤2：N,N-二甲基-4-(4-氧代环己基)苯磺酰胺的合成Step 2: Synthesis of N,N-dimethyl-4-(4-oxocyclohexyl)benzenesulfonamide

N,N-二甲基-4-{1,4-二氧杂螺[4.5]癸-8-基}苯磺酰胺(600mg,1.85mmol,1.0eq)、对甲基苯磺酸水合物(527mg,2.77mmol,1.5eq)溶于丙酮(9mL)和水(3mL)中，60℃反应2小时。待反应完全，混合液用碳酸氢钠溶液淬灭，再用乙酸乙酯和水萃取，有机相用无水硫酸钠除去多余水分，浓缩、旋干，得到黄色固体N,N-二甲基-4-(4-氧代环己基)苯磺酰胺(530mg)，直接用于下一步反应。LCMS(ESI)[M+H]:282.1233。1H NMR(400MHz,DMSO)δ7.72–7.65(m,2H),7.58(d,J＝8.0Hz,2H),3.20(m,J＝12.1,3.5Hz,1H),2.60(d,J＝1.3Hz,8H),2.29(d,J＝13.8Hz,2H),2.10(m,J＝13.1,6.4,3.4Hz,2H),2.01–1.85(m,2H)。N,N-Dimethyl-4-{1,4-dioxaspiro[4.5]dec-8-yl}benzenesulfonamide (600 mg, 1.85 mmol, 1.0 eq) and p-toluenesulfonic acid hydrate (527 mg, 2.77 mmol, 1.5 eq) were dissolved in acetone (9 mL) and water (3 mL) and reacted at 60°C for 2 hours. Upon completion, the mixture was quenched with sodium bicarbonate solution and extracted with ethyl acetate and water. The organic phase was dehydrated with anhydrous sodium sulfate, concentrated, and dried to afford a yellow solid, N,N-dimethyl-4-(4-oxocyclohexyl)benzenesulfonamide (530 mg), which was used directly in the next reaction. LCMS (ESI) [M+H]: 282.1233. 1H NMR (400MHz, DMSO) δ7.72–7.65(m,2H),7.58(d,J＝8.0Hz,2H),3.20(m,J＝12.1,3.5Hz,1H),2.6 0(d,J＝1.3Hz,8H),2.29(d,J＝13.8Hz,2H),2.10(m,J＝13.1,6.4,3.4Hz,2H),2.01–1.85(m,2H).

步骤3：4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-N,N-二甲基苯磺酰胺的合成Step 3: Synthesis of 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-N,N-dimethylbenzenesulfonamide

将N、N-二甲基-4-(4-氧代环己基)苯磺酰胺(150mg,0.534mmol,1.0eq)、硫脲(81.3mg,1.07mmol,2.0eq)和碘单质(163mg,0.641mmol,1.2eq)溶于乙醇(10mL)中，90℃下反应16小时。反应完全后，将混合液浓缩，然后用饱和的碳酸氢钠溶液和硫代硫酸钠溶液淬灭，再用乙酸乙酯和水萃取，得到黄色固体4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-N,N-二甲基苯磺酰胺(150mg,收率83.3％)。LCMS(ESI)[M+H]:338.1068。1H NMR(400MHz,DMSO)δ7.68(d,J＝7.8Hz,2H),7.58(dd,J＝8.4,2.2Hz,2H),2.60(d,J＝1.6Hz,7H),2.50(s,3H),2.33–2.24(m,1H),2.10(d,J＝11.9Hz,1H),2.03–1.83(m,2H)。N,N-Dimethyl-4-(4-oxocyclohexyl)benzenesulfonamide (150 mg, 0.534 mmol, 1.0 eq), thiourea (81.3 mg, 1.07 mmol, 2.0 eq), and iodine (163 mg, 0.641 mmol, 1.2 eq) were dissolved in ethanol (10 mL) and reacted at 90°C for 16 hours. After the reaction was complete, the mixture was concentrated and quenched with saturated sodium bicarbonate and sodium thiosulfate solutions. The mixture was then extracted with ethyl acetate and water to afford 4-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-N,N-dimethylbenzenesulfonamide (150 mg, 83.3% yield) as a yellow solid. LCMS (ESI) [M+H]: 338.1068. 1H NMR (400MHz, DMSO) δ7.68 (d, J = 7.8 Hz, 2H), 7.58 (dd, J = 8.4, 2.2 Hz, 2H), 2.60 (d, J = 1. 6Hz, 7H), 2.50 (s, 3H), 2.33–2.24 (m, 1H), 2.10 (d, J = 11.9Hz, 1H), 2.03–1.83 (m, 2H).

步骤4：2'-氯-N-{6-[4-(N,N-二甲基氨磺酰基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 4: Synthesis of 2'-chloro-N-{6-[4-(N,N-dimethylsulfamoyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将4-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-N,N-二甲基苯磺酰胺(135mg,0.40mmol,1.0eq)、2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(111mg,0.40mmol,1.0eq)、N,N,N,N-四甲基氯甲眯六氟磷酸盐(168mg,0.60mmol,1.5eq)和N-甲基咪唑(115mg,1.41mmol,3.5eq)溶于超干的N,N-二甲基甲酰胺(3.5mL)中，室温下反应2小时。反应完全后，制备纯化得到淡黄色固体2'-氯-N-{6-[4-(N,N-二甲基氨磺酰基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(10.0mg,收率4.2％)。LCMS(ESI)[M+H]:598.1321。1H NMR(400MHz,DMSO)δ12.48(s,1H),8.79(s,1H),8.17(s,1H),7.70(d,J＝8.1Hz,2H),7.62(d,J＝8.2Hz,2H),7.50(s,1H),7.39(s,1H),3.62(s,3H),3.00–2.69(m,5H),2.59(d,J＝11.7Hz,9H),2.07(t,J＝7.4Hz,2H)。4-(2-Amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-N,N-dimethylbenzenesulfonamide (135 mg, 0.40 mmol, 1.0 eq), 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (111 mg, 0.40 mmol, 1.0 eq), N,N,N,N-tetramethylchloromethylimidazole hexafluorophosphate (168 mg, 0.60 mmol, 1.5 eq) and N-methylimidazole (115 mg, 1.41 mmol, 3.5 eq) were dissolved in ultra-dry N,N-dimethylformamide (3.5 mL) and reacted at room temperature for 2 hours. After the reaction was complete, the product was purified and prepared to give 2'-chloro-N-{6-[4-(N,N-dimethylsulfamoyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (10.0 mg, 4.2% yield) as a pale yellow solid. LCMS (ESI) [M+H]: 598.1321. 1H NMR (400MHz, DMSO) δ12.48(s,1H),8.79(s,1H),8.17(s,1H),7.70(d,J＝8.1Hz,2H),7.62(d,J＝8.2Hz,2H) ,7.50(s,1H),7.39(s,1H),3.62(s,3H),3.00–2.69(m,5H),2.59(d,J＝11.7Hz,9H),2.07(t,J＝7.4Hz,2H).

实施例34：2'-氯-5'-甲氧基-6-甲基-N-(6-(6-((2,2,2-三氟乙基)氨基)吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(化合物56)的合成
Example 34: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-(6-(6-((2,2,2-trifluoroethyl)amino)pyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (Compound 56)

步骤1：5-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)吡啶-2-胺的合成Step 1: Synthesis of 5-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyridin-2-amine

将4,4,5,5-四甲基-2-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)-1,3,2-二氧杂硼烷(5.0g,18.79mmol,1.0eq)和5-溴吡啶-2-胺(3.25g,18.79mmol,1.0eq)溶于1,4-二氧六环(50.0mL)和水(12.5mL)的混合溶液中，加入碳酸铯(12.25g,37.58mmol,2.0eq)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(274mg,0.375mmol,0.03eq)，氮气保护下，80℃反应2小时。待反应完全，用乙酸乙酯萃取，正相纯化(石油醚/乙酸乙酯＝3:1)得棕色固体5-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)吡啶-2-胺(4.07g,收率93.34％)。LCMS(ESI)[M+H]+:233.12。4,4,5,5-Tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (5.0 g, 18.79 mmol, 1.0 eq) and 5-bromopyridin-2-amine (3.25 g, 18.79 mmol, 1.0 eq) were dissolved in a mixed solution of 1,4-dioxane (50.0 mL) and water (12.5 mL). Cesium carbonate (12.25 g, 37.58 mmol, 2.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (274 mg, 0.375 mmol, 0.03 eq) were added, and the mixture was reacted at 80°C under nitrogen protection for 2 hours. After the reaction was complete, the product was extracted with ethyl acetate and purified by normal phase purification (petroleum ether/ethyl acetate = 3:1) to give 5-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyridin-2-amine (4.07 g, yield 93.34%) as a brown solid. LCMS (ESI) [M+H]+: 233.12.

步骤2：5-(1,4-二氧阿司匹林[4.5]癸-8-基)吡啶-2-胺的合成Step 2: Synthesis of 5-(1,4-dioxyaspirino[4.5]dec-8-yl)pyridin-2-amine

将5-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)吡啶-2-胺(4.0g,17.2mmol,1.0eq)溶于甲醇(30mL)中，加入钯/碳(183mg,1.72mmol,0.1eq)，随后氢气氛围下，室温反应8小时。待反应完全，将反应液过滤，滤液浓缩得黄色固体粗品5-(1,4-二氧阿司匹林[4.5]癸-8-基)吡啶-2-胺(4.0g)，直接用于下一步反应。LCMS(ESI)[M+H]+:235.1。1H NMR(400MHz,DMSO)δ8.88(s,1H),7.48(s,1H),4.04(s,3H)。5-(1,4-Dioxaspiro[4.5]dec-7-en-8-yl)pyridin-2-amine (4.0 g, 17.2 mmol, 1.0 eq) was dissolved in methanol (30 mL) and palladium/carbon (183 mg, 1.72 mmol, 0.1 eq) was added. The mixture was then reacted at room temperature under a hydrogen atmosphere for 8 hours. Upon completion, the reaction solution was filtered and the filtrate concentrated to afford crude 5-(1,4-dioxaspirino[4.5]dec-8-yl)pyridin-2-amine (4.0 g) as a yellow solid, which was used directly in the next step. LCMS (ESI) [M+H]+: 235.1. 1H NMR (400 MHz, DMSO) δ 8.88 (s, 1H), 7.48 (s, 1H), 4.04 (s, 3H).

步骤3：4-(6-氨基吡啶-3-基)环己-1-酮的合成Step 3: Synthesis of 4-(6-aminopyridin-3-yl)cyclohexan-1-one

将5-(1,4-二氧阿司匹林[4.5]癸-8-基)吡啶-2-胺(2.0g,8.6mmol,1.0eq)溶于四氢呋喃(10.0mL)和乙醇5.0mL)混合溶液中，随后加入浓盐酸(5.0mL)，室温反应2小时。待反应完全，调节pH至中性，正相纯化(二氯甲烷/甲醇＝10：1)得到黄色固体4-(6-氨基吡啶-3-基)环己-1-酮(1.31g,收率80.36％)。LCMS(ESI)[M+H]+:190.1。5-(1,4-Dioxyaspirino[4,5]dec-8-yl)pyridin-2-amine (2.0 g, 8.6 mmol, 1.0 eq) was dissolved in a mixture of tetrahydrofuran (10.0 mL) and ethanol (5.0 mL). Concentrated hydrochloric acid (5.0 mL) was then added and allowed to react at room temperature for 2 hours. Upon completion of the reaction, the pH was adjusted to neutral and the product was purified by normal phase purification (dichloromethane/methanol = 10:1) to afford 4-(6-aminopyridin-3-yl)cyclohexan-1-one (1.31 g, 80.36% yield) as a yellow solid. LCMS (ESI) [M+H]+: 190.1.

步骤4：2,2,2-三氟-N-[5-(4-氧代环己基)吡啶-2-基]乙酰胺的合成Step 4: Synthesis of 2,2,2-trifluoro-N-[5-(4-oxocyclohexyl)pyridin-2-yl]acetamide

将4-(6-氨基吡啶-3-基)环己-1-酮(1.3g,6.8mmol,1.0eq)溶于超干二氯甲烷(25mL)中，冰浴条件下，依次滴加三乙胺(2.06g,20.41mmol,3.0eq)和三氟乙酸酐(2.14g,10.2mmol,1.5eq)，随后室温反应1.5小时。待反应完全，经正相(石油醚:乙酸乙酯＝3:2)纯化得到白色固体2,2,2-三氟-N-[5-(4-氧代环己基)吡啶-2-基]乙酰胺(1.41g,收率72.68％)。LCMS(ESI)[M+H]+:287.1。4-(6-Aminopyridin-3-yl)cyclohexan-1-one (1.3 g, 6.8 mmol, 1.0 eq) was dissolved in ultra-dry dichloromethane (25 mL). Triethylamine (2.06 g, 20.41 mmol, 3.0 eq) and trifluoroacetic anhydride (2.14 g, 10.2 mmol, 1.5 eq) were added dropwise in an ice bath. The mixture was allowed to react at room temperature for 1.5 hours. Upon completion, the reaction was purified by normal phase chromatography (petroleum ether:ethyl acetate = 3:2) to afford 2,2,2-trifluoro-N-[5-(4-oxocyclohexyl)pyridin-2-yl]acetamide (1.41 g, 72.68% yield) as a white solid. LCMS (ESI) [M + H] + : 287.1.

步骤5：N-[5-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)吡啶-2-基]-2,2,2-三氟乙酰胺的合成Step 5: Synthesis of N-[5-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)pyridin-2-yl]-2,2,2-trifluoroacetamide

将2,2,2-三氟-N-[5-(4-氧代环己基)吡啶-2-基]乙酰胺(400mg,1.39mmol,1.0eq)溶四氢呋喃中，加苯基三甲基三溴化铵(522.5mg,1.39mmol,1.0eq)，室温反应2小时，随后用乙酸乙酯萃取分液浓缩，用乙腈溶解，加入硫脲(116.2mg,1.52mmol,1.1eq)，室温继续反应4小时。待反应结束浓缩得黄色液体粗品N-[5-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)吡啶-2-基]-2,2,2-三氟乙酰胺(677mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:343.08。2,2,2-Trifluoro-N-[5-(4-oxocyclohexyl)pyridin-2-yl]acetamide (400 mg, 1.39 mmol, 1.0 eq) was dissolved in tetrahydrofuran and phenyltrimethylammonium tribromide (522.5 mg, 1.39 mmol, 1.0 eq) was added. The mixture was allowed to react at room temperature for 2 hours. The mixture was then extracted with ethyl acetate, concentrated, and dissolved in acetonitrile. Thiourea (116.2 mg, 1.52 mmol, 1.1 eq) was added and the reaction continued at room temperature for 4 hours. After completion of the reaction, the mixture was concentrated to yield a crude yellow liquid, N-[5-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)pyridin-2-yl]-2,2,2-trifluoroacetamide (677 mg), which was used directly in the next reaction. LCMS (ESI) [M + H] + : 343.08.

步骤6：6-{6-[(2,2,2-三氟乙基)氨基]吡啶-3-基}-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 6: Synthesis of 6-{6-[(2,2,2-trifluoroethyl)amino]pyridin-3-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将硼氢化钠(200mg,5.28mmol,1.0eq)和氯化锌(1.44g,10.57mmol,2.0eq)溶于超干四氢呋喃中(20mL)中，室温反应过夜，得到体系1溶液，另称取N-[5-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)吡啶-2-基]-2,2,2-三氟乙酰胺(300mg,0.876mmol,1.0eq)溶于超过干四氢呋喃(10.0mL)中，冰浴条件下，缓慢滴加体系1溶液，80℃反应2小时，加入甲醇萃取分液，浓缩得黄色液体粗品6-{6-[(2,2,2-三氟乙基)氨基]吡啶-3-基}-4,5,6,7-四氢苯并[d]噻唑-2-胺(533mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:329.1。Sodium borohydride (200 mg, 5.28 mmol, 1.0 eq) and zinc chloride (1.44 g, 10.57 mmol, 2.0 eq) were dissolved in ultra-dry tetrahydrofuran (20 mL) and reacted at room temperature overnight to obtain a solution of System 1. N-[5-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)pyridin-2-yl]-2,2,2-trifluoroacetamide (300 mg) was weighed. , 0.876 mmol, 1.0 eq) was dissolved in an excess of dry tetrahydrofuran (10.0 mL). System 1 solution was slowly added dropwise under ice-cooling conditions. The mixture was allowed to react at 80°C for 2 hours. Methanol was added for extraction and the separated liquid was concentrated to afford a crude yellow liquid, 6-{6-[(2,2,2-trifluoroethyl)amino]pyridin-3-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (533 mg), which was used directly in the next reaction. LCMS (ESI) [M + H] + : 329.1.

步骤7：2'-氯-5'-甲氧基-6-甲基-N-(6-((2,2,2-三氟乙基)氨基)吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺的合成Step 7: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-(6-((2,2,2-trifluoroethyl)amino)pyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide

将6-{6-[(2,2,2-三氟乙基)氨基]吡啶-3-基}-4,5,6,7-四氢苯并[d]噻唑-2-胺(100.0mg,0.304mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(84.7mg,0.304mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(2mL)中，加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(126mg,0.456mmol,1.5eq)和N-甲基咪唑(74.8mg,0.912mmol,3.0eq)，室温反应1.5小时。反应结束后，制备纯化得2'-氯-5'-甲氧基-6-甲基-N-(6-((2,2,2-三氟乙基)氨基)吡啶-3-基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(16.3mg,收率9.10％)。LCMS(ESI)[M+H]+:589.1。6-{6-[(2,2,2-trifluoroethyl)amino]pyridin-3-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (100.0 mg, 0.304 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (84.7 mg, 0.304 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (2 mL). N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (126 mg, 0.456 mmol, 1.5 eq) and N-methylimidazole (74.8 mg, 0.912 mmol, 3.0 eq) were added and reacted at room temperature for 1.5 hours. After the reaction, the product was purified and prepared to give 2'-chloro-5'-methoxy-6-methyl-N-(6-((2,2,2-trifluoroethyl)amino)pyridin-3-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (16.3 mg, 9.10% yield). LCMS (ESI) [M + H]+: 589.1.

实施例35：2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(N-乙基氨基磺酰基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(化合物62)的合成
Example 35: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(N-ethylaminosulfonyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (Compound 62)

步骤1：4-溴-N-乙苯磺酰胺的合成Step 1: Synthesis of 4-bromo-N-ethylbenzenesulfonamide

将4-溴苯磺酰氯(25.5g,0.1mol,1.0eq)、乙胺盐酸盐(9.78g,0.12mol,1.2eq)和三乙胺(30.3g,0.3mmol,3.0eq)溶于二氯甲烷中，室温下搅拌反应0.5小时。反应完全后，加入二氯甲烷和水萃取分液，合并有机相，浓缩后得到白色固体4-溴-N-乙苯磺酰胺(26.6g)，直接用于下一步反应。LCMS(ESI)[M+H]:263.9。1H NMR(400MHz,DMSO-d6)δ7.84–7.79(m,2H),7.73–7.69(m,2H),7.67(t,J＝5.7Hz,1H),2.78(m,J＝7.2,5.6Hz,2H),0.97(t,J＝7.2Hz,3H)。4-Bromobenzenesulfonyl chloride (25.5 g, 0.1 mol, 1.0 eq), ethylamine hydrochloride (9.78 g, 0.12 mol, 1.2 eq), and triethylamine (30.3 g, 0.3 mmol, 3.0 eq) were dissolved in dichloromethane and stirred at room temperature for 0.5 hour. After the reaction was complete, dichloromethane and water were added to extract the separated liquids. The organic phases were combined and concentrated to obtain 4-bromo-N-ethylbenzenesulfonamide (26.6 g) as a white solid, which was used directly in the next reaction. LCMS (ESI) [M+H]: 263.9. 1 H NMR (400MHz, DMSO-d6) δ7.84–7.79(m,2H),7.73–7.69(m,2H),7.67(t,J＝5.7Hz,1H),2.78(m,J＝7.2,5.6Hz,2H),0.97(t,J＝7.2Hz,3H).

步骤2：N-乙基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯磺酰胺的合成Step 2: Synthesis of N-ethyl-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzenesulfonamide

将4-溴-N-乙基苯磺酰胺(26.3g,0.1mol,1.0eq)、4,4,5,5-四甲基-2-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)-1,3,2-二氧硼烷(26.6g,0.1mol,1.0eq)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(7.3g,0.01mol,0.1eq)、碳酸钾(41g,0.3mol,3.0eq)溶于1,4-二氧六环(300mL)和水(50mL)中，氮气保护体系，80℃下反应3小时。反应完全后，过滤反应液，浓缩、拌样，然后正相柱分离纯化(乙酸乙酯/石油醚，乙酸乙酯％＝60％)，得到黄色固体N-乙基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯磺酰胺(22.82g,收率70.43％)。LCMS(ESI)[M+H]:324。1H NMR(400MHz,DMSO-d6)δ7.71(d,J＝8.4Hz,2H),7.62(d,J＝8.4Hz,2H),7.51(t,J＝5.7Hz,1H),6.30–6.14(m,1H),3.92(s,4H),2.80–2.71(m,2H),2.57(m,J＝4.6,2.4Hz,2H),2.40(m,J＝4.3,2.4Hz,2H),1.82(t,J＝6.5Hz,2H),0.96(t,J＝7.2Hz,3H)。4-Bromo-N-ethylbenzenesulfonamide (26.3 g, 0.1 mol, 1.0 eq), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (26.6 g, 0.1 mol, 1.0 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (7.3 g, 0.01 mol, 0.1 eq), and potassium carbonate (41 g, 0.3 mol, 3.0 eq) were dissolved in 1,4-dioxane (300 mL) and water (50 mL), and the system was protected by nitrogen. The reaction was carried out at 80 ° C for 3 hours. After the reaction was complete, the reaction solution was filtered, concentrated, and stirred, then purified by normal phase column chromatography (ethyl acetate/petroleum ether, ethyl acetate % = 60%) to obtain N-ethyl-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzenesulfonamide (22.82 g, yield 70.43%) as a yellow solid. LCMS (ESI) [M+H]: 324. 1H NMR (400MHz, DMSO-d6) δ7.71(d,J＝8.4Hz,2H),7.62(d,J＝8.4Hz,2H),7.51(t,J＝5.7Hz,1H),6.30–6.14(m,1H),3.92(s,4H ), 2.80–2.71 (m, 2H), 2.57 (m, J = 4.6, 2.4Hz, 2H), 2.40 (m, J = 4.3, 2.4Hz, 2H), 1.82 (t, J = 6.5Hz, 2H), 0.96 (t, J = 7.2Hz, 3H).

步骤3：N-乙基-4-(1,4-二氧杂螺[4.5]癸烷-8-基)苯磺酰胺的合成Step 3: Synthesis of N-ethyl-4-(1,4-dioxaspiro[4.5]decan-8-yl)benzenesulfonamide

将N-乙基-4-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)苯磺酰胺(17g,52.63mmol,1.0eq)、钯碳(1.7g)溶于甲醇(200mL)中，氢气置换三次，然后室温下反应2.5小时。反应完全后，将反应液过滤，滤液浓缩后得到黄色油状物N-乙基-4-(1,4-二氧杂螺[4.5]癸烷-8-基)苯磺酰胺(14.66g,收率85.73％)。LCMS(ESI)[M+H]+:326。1H NMR(400MHz,DMSO-d6)δ7.79–7.63(m,2H),7.46(dd,J＝7.0,5.1Hz,3H),3.91(d,J＝2.1Hz,4H),2.84–2.69(m,2H),1.82–1.56(m,9H),0.98(t,J＝7.2Hz,3H)。N-ethyl-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)benzenesulfonamide (17 g, 52.63 mmol, 1.0 eq) and palladium on carbon (1.7 g) were dissolved in methanol (200 mL) and the atmosphere was replaced with hydrogen three times. The mixture was then allowed to react at room temperature for 2.5 hours. After the reaction was complete, the reaction solution was filtered and the filtrate was concentrated to afford N-ethyl-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzenesulfonamide (14.66 g, 85.73% yield) as a yellow oil. LCMS (ESI) [M+H]+: 326. 1H NMR(400MHz, DMSO-d6)δ7.79–7.63(m,2H),7.46(dd,J＝7.0,5.1Hz,3H),3.91( d,J＝2.1Hz,4H),2.84–2.69(m,2H),1.82–1.56(m,9H),0.98(t,J＝7.2Hz,3H).

步骤4：N-乙基-4-(4-氧代环己基)苯磺酰胺的合成Step 4: Synthesis of N-ethyl-4-(4-oxocyclohexyl)benzenesulfonamide

N-乙基-4-(1,4-二氧杂螺[4.5]癸烷-8-基)苯磺酰胺(14.66g,45.11mmol,1.0eq)、对甲基苯磺酸水合物(8.57mg,45.11mmol,1.0eq)溶于丙酮(50mL)和水(100mL)中，50℃反应4小时。反应完全后，混合液用碳酸氢钠溶液淬灭，再用乙酸乙酯和水萃取，有机相用无水硫酸钠除去多余水分，浓缩得到黄色油状物N-乙基-4-(4-氧代环己基)苯磺酰胺(9.1mg,收率91％)。LCMS(ESI)[M+H]:282。N-Ethyl-4-(1,4-dioxaspiro[4.5]dec-8-yl)benzenesulfonamide (14.66 g, 45.11 mmol, 1.0 eq) and p-toluenesulfonic acid hydrate (8.57 mg, 45.11 mmol, 1.0 eq) were dissolved in acetone (50 mL) and water (100 mL) and reacted at 50°C for 4 hours. After the reaction was complete, the mixture was quenched with sodium bicarbonate solution and extracted with ethyl acetate and water. The organic phase was dehydrated with anhydrous sodium sulfate and concentrated to afford N-ethyl-4-(4-oxocyclohexyl)benzenesulfonamide (9.1 mg, 91% yield) as a yellow oil. LCMS (ESI) [M+H]: 282.

步骤5：4-(2-氨基-4,5,6,7-四氢苯并噻唑-6-基)-N-乙基苯磺酰胺的合成Step 5: Synthesis of 4-(2-amino-4,5,6,7-tetrahydrobenzothiazol-6-yl)-N-ethylbenzenesulfonamide

将N-乙基-4-(4-氧代环己基)苯磺酰胺(9.1g,32.38mmol,1.0eq)、硫脲(4.9g,64.79mmol,2.0eq)和碘单质(12.3g,48.57mmol,1.5eq)溶于乙醇(200mL)中，80℃下反应2小时。反应完全后，将反应混合液浓缩，然后用饱和的碳酸氢钠溶液和硫代硫酸钠溶液淬灭，再用乙酸乙酯和水萃取，有机相浓缩得到黄色固体粗品4-(2-氨基-4,5,6,7-四氢苯并噻唑-6-基)-N-乙基苯磺酰胺(5.85g)。N-ethyl-4-(4-oxocyclohexyl)benzenesulfonamide (9.1 g, 32.38 mmol, 1.0 eq), thiourea (4.9 g, 64.79 mmol, 2.0 eq), and iodine (12.3 g, 48.57 mmol, 1.5 eq) were dissolved in ethanol (200 mL) and reacted at 80°C for 2 hours. After the reaction was complete, the reaction mixture was concentrated and then quenched with saturated sodium bicarbonate solution and sodium thiosulfate solution. The mixture was then extracted with ethyl acetate and water, and the organic phase was concentrated to obtain a crude yellow solid of 4-(2-amino-4,5,6,7-tetrahydrobenzothiazol-6-yl)-N-ethylbenzenesulfonamide (5.85 g).

步骤6：2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(N-乙基氨基磺酰基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(N-ethylaminosulfonyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide

将4-(2-氨基-4,5,6,7-四氢苯并噻唑-6-基)-N-乙基苯磺酰胺(3g,8.9mmol,1.0eq)、2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(2.5g,8.9mmol,1.0eq)、N,N,N,N-四甲基氯甲眯六氟磷酸盐(3.7g,13.4mmol,1.5eq)、N-甲基咪唑(2.2g,26.7mmol,3.0eq)溶于乙腈(100mL)中，室温下反应2小时。反应完全后，正向柱分离，纯乙酸乙酯出产物，再经过反向柱分离纯化，乙腈占90％时流出产物，浓缩得到黄色粉末2'-氯-5'-甲氧基-6-甲基-N-{6-[4-(N-乙基氨基磺酰基)苯基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(1.77g,收率33.21％)。LCMS(ESI)[M+H]:598.1329。1H NMR(400MHz,DMSO-d6)δ12.54(s,1H),8.78(s,1H),8.17(s,1H),7.78–7.68(m,2H),7.56(d,J＝8.3Hz,2H),7.53–7.47(m,2H),7.41(s,1H),3.62(s,3H),3.14(m,J＝10.3,4.9Hz,1H),2.96(dd,J＝15.8,5.0Hz,1H),2.84–2.70(m,5H),2.58(s,3H),2.09–1.99(m,2H),0.97(t,J＝7.2Hz,3H)。4-(2-Amino-4,5,6,7-tetrahydrobenzothiazol-6-yl)-N-ethylbenzenesulfonamide (3 g, 8.9 mmol, 1.0 eq), 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (2.5 g, 8.9 mmol, 1.0 eq), N,N,N,N-tetramethylchloromethylimidazole hexafluorophosphate (3.7 g, 13.4 mmol, 1.5 eq), and N-methylimidazole (2.2 g, 26.7 mmol, 3.0 eq) were dissolved in acetonitrile (100 mL) and reacted at room temperature for 2 hours. After the reaction was complete, the product was separated by a forward column and purified by pure ethyl acetate. The product was then separated and purified by a reverse column. When the acetonitrile content was 90%, the product was concentrated to afford a yellow powder, 2'-chloro-5'-methoxy-6-methyl-N-{6-[4-(N-ethylaminosulfonyl)phenyl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (1.77 g, 33.21% yield). LCMS (ESI) [M+H]: 598.1329. 1H NMR (400MHz, DMSO-d6) δ12.54(s,1H),8.78(s,1H),8.17(s,1H),7.78–7.68(m,2H),7.56(d,J＝8.3Hz,2H),7.53–7.47(m,2H),7.41(s,1H),3.6 2(s,3H),3.14(m,J＝10.3,4.9Hz,1H),2.96(dd,J＝15.8,5.0Hz,1H),2.84–2.70(m,5H),2.58(s,3H),2.09–1.99(m,2H),0.97(t,J＝7.2Hz,3H).

实施例36：2'-氯-5'-甲氧基-6-甲基-N-(6-(4-(2,2,2-三氟乙基)氨基)哌啶-1-基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(化合物65)的合成
Example 36: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-(6-(4-(2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (Compound 65)

步骤1：4-(2,2,2-三氟乙基)氨基)哌啶-1-甲酸叔丁酯的合成Step 1: Synthesis of tert-butyl 4-(2,2,2-trifluoroethyl)amino)piperidine-1-carboxylate

将4-氯哌啶-1-甲酸叔丁酯(5g,23.0mmol,1.0eq)溶于二氯甲烷(20ml)中，再加入2,2,2-三氟乙基胺(2.7g,27.0mmol,1.2eq)和三乙胺(6.9g,69.0mol,3.0eq)，在室温下反应2小时。待反应完全，加入水和乙酸乙酯(100mL*3)萃取分液，有机相干燥、浓缩得棕色油状物4-(2,2,2-三氟乙基)氨基)哌啶-1-甲酸叔丁酯(4.1g,收率63.2％)。LCMS(ESI)[M+H]+:283.1。Dissolve tert-butyl 4-chloropiperidine-1-carboxylate (5 g, 23.0 mmol, 1.0 eq) in dichloromethane (20 ml), then add 2,2,2-trifluoroethylamine (2.7 g, 27.0 mmol, 1.2 eq) and triethylamine (6.9 g, 69.0 mol, 3.0 eq). Allow to react at room temperature for 2 hours. After the reaction is complete, extract with water and ethyl acetate (100 mL x 3). The organic phase is dried and concentrated to yield tert-butyl 4-(2,2,2-trifluoroethyl)amino)piperidine-1-carboxylate (4.1 g, 63.2% yield) as a brown oil. LCMS (ESI) [M+H]+: 283.1.

步骤2：N-(2,2,2-三氟乙基)哌啶-4-胺的合成Step 2: Synthesis of N-(2,2,2-trifluoroethyl)piperidin-4-amine

将4-(2,2,2-三氟乙基)氨基)哌啶-1-甲酸叔丁酯(4.1g,14.0mmol,1.0eq)溶于二氯甲烷(20mL)中，再缓慢加入三氟乙酸(5mL)，在室温下反应2小时。待反应完全，加入水和乙酸乙酯(100mL*3)萃取分液，有机相浓缩、干燥得棕色油状物N-(2,2,2-三氟乙基)哌啶-4-胺(2.1g,收率82.4％)。LCMS(ESI)[M+H]+:183.1。Dissolve tert-butyl 4-(2,2,2-trifluoroethyl)amino)piperidin-1-carboxylate (4.1 g, 14.0 mmol, 1.0 eq) in dichloromethane (20 mL). Slowly add trifluoroacetic acid (5 mL) and allow to react at room temperature for 2 hours. Upon completion, extract with water and ethyl acetate (100 mL x 3). The organic phase is concentrated and dried to afford N-(2,2,2-trifluoroethyl)piperidin-4-amine (2.1 g, 82.4% yield) as a brown oil. LCMS (ESI) [M+H]+: 183.1.

步骤3：(6-(4-(2,2,2-三氟乙基)氨基)哌啶-1-基)-4,5,6,7-四氢苯并[d]噻唑-2-基)氨基甲酸叔丁酯的合成Step 3: Synthesis of tert-butyl (6-(4-(2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)carbamate

将N-(2,2,2-三氟乙基)哌啶-4-胺(2.0g,11.0mmol,1.0eq)溶于甲醇(20mL)中，加入(5-氧代-4,5,6,7-四氢苯并[d]噻唑-2-基)氨基甲酸叔丁酯(3.24g,12.0mmol,1.1eq)、氰基硼氢化钠(2.07g,33.0mmol,3.0eq)和醋酸(660g,11.0mmol,1.0eq)，80℃反应4小时。待反应完全后，经正相(石油醚:乙酸乙酯＝2:1)纯化，得棕色固体(6-(4-(2,2,2-三氟乙基)氨基)哌啶-1-基)-4,5,6,7-四氢苯并[d]噻唑-2-基)氨基甲酸叔丁酯(600mg,收率12.5％)。LCMS(ESI)[M+H]+:435.2。Dissolve N-(2,2,2-trifluoroethyl)piperidin-4-amine (2.0 g, 11.0 mmol, 1.0 eq) in methanol (20 mL), add tert-butyl (5-oxo-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)carbamate (3.24 g, 12.0 mmol, 1.1 eq), sodium cyanoborohydride (2.07 g, 33.0 mmol, 3.0 eq), and acetic acid (660 g, 11.0 mmol, 1.0 eq). The mixture was reacted at 80°C for 4 hours. After completion of the reaction, the product was purified by normal phase chromatography (petroleum ether:ethyl acetate = 2:1) to afford tert-butyl (6-(4-(2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)carbamate (600 mg, 12.5% yield) as a brown solid. LCMS (ESI) [M+H] +: 435.2.

步骤4：6-(4-((2,2,2-三氟乙基)氨基)哌啶-1-基)-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 4: Synthesis of 6-(4-((2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将(6-(4-(2,2,2-三氟乙基)氨基)哌啶-1-基)-4,5,6,7-四氢苯并[d]噻唑-2-基)氨基甲酸叔丁酯(600mg,1.38mmol,1.0eq)溶于二氯甲挖(8mL)中，再缓慢加入三氟乙酸(1mL)，在室温下反应30分钟。待反应完全，加入水和乙酸乙酯(100mL*3)萃取分液，有机相浓缩、干燥得棕色油状物6-(4-((2,2,2-三氟乙基)氨基)哌啶-1-基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(310mg,收率67.2％)。LCMS(ESI)[M+H]+:335.14.Dissolve tert-butyl (6-(4-(2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)carbamate (600 mg, 1.38 mmol, 1.0 eq) in dichloromethane (8 mL). Slowly add trifluoroacetic acid (1 mL) and react at room temperature for 30 minutes. After the reaction is complete, extract with water and ethyl acetate (100 mL x 3). The organic phase is concentrated and dried to yield 6-(4-((2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine as a brown oil (310 mg, 67.2% yield). LCMS (ESI) [M+H]+: 335.14.

步骤5：2'-氯-5'-甲氧基-6-甲基-N-(6-(4-(2,2,2-三氟乙基)氨基)哌啶-1-基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺的合成Step 5: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-(6-(4-(2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide

将6-(4-((2,2,2-三氟乙基)氨基)哌啶-1-基)-4,5,6,7-四氢苯并[d]噻唑-2-胺(300mg,0.90mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(249mg,0.90mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(4mL)中，加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(378mg,1.3mmol,1.5eq)和N-甲基咪唑(221mg,2.7mmol,3.0eq)，随后室温反应5小时。待反应完全，制备得白色固体2'-氯-5'-甲氧基-6-甲基-N-(6-(4-(2,2,2-三氟乙基)氨基)哌啶-1-基)-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(20mg,收率3.7％)。LCMS(ESI)[M+H]+:595.1。6-(4-((2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (300 mg, 0.90 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (249 mg, 0.90 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (4 mL), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (378 mg, 1.3 mmol, 1.5 eq) and N-methylimidazole (221 mg, 2.7 mmol, 3.0 eq) were added, followed by reaction at room temperature for 5 hours. After the reaction was complete, a white solid, 2'-chloro-5'-methoxy-6-methyl-N-(6-(4-(2,2,2-trifluoroethyl)amino)piperidin-1-yl)-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (20 mg, 3.7% yield) was obtained. LCMS (ESI) [M+H]+: 595.1.

实施例37：2'-氯-5'-甲氧基-6-甲基-N-{6-[(1-甲基氮杂环丁烷-3-基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(化合物68)的合成
Example 37: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[(1-methylazetidin-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (Compound 68)

步骤1：3-(氯甲基)-1-甲基氮杂环丁烷的合成Step 1: Synthesis of 3-(chloromethyl)-1-methylazetidine

将3-(氯甲基)氮杂环丁烷(3g,28.0mmol,1.0eq)溶于二氯甲烷(20mL)中，再加入碘甲烷(4.8g,34.0mmol,1.2eq)、三乙胺(8.6g,0.086mol,3.0eq)，在50℃下反应2小时。待反应完全，加入水和乙酸乙酯(100mL*3)萃取分液，有机相干燥、浓缩得棕色固体3-(氯甲基)-1-甲基氮杂环丁烷(1.8g,收率54％)。LCMS(ESI)[M+H]+:120.05。Dissolve 3-(Chloromethyl)azetidine (3 g, 28.0 mmol, 1.0 eq) in dichloromethane (20 mL), then add iodomethane (4.8 g, 34.0 mmol, 1.2 eq) and triethylamine (8.6 g, 0.086 mol, 3.0 eq). Incubate at 50°C for 2 hours. After completion, extract with water and ethyl acetate (100 mL x 3). The organic phase is dried and concentrated to yield a brown solid, 3-(chloromethyl)-1-methylazetidine (1.8 g, 54% yield). LCMS (ESI) [M+H]+: 120.05.

步骤2：6-[(1-甲基氮杂环丁烷-3-基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 2: Synthesis of 6-[(1-methylazetidin-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将3-(氯甲基)-1-甲基氮杂环丁烷(1.2g,10mmol,1.0eq)溶于无水乙醇(10mL)中，再加入2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-醇(1.7g,10.0mmol,1.0eq)、碳酸钾(4.1g,30.0mmol,3.0eq)，80℃下反应5小时。待反应完全，反向柱纯化得棕色油状物6-[(1-甲基氮杂环丁烷-3-基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(560mg,收率22.1％)。LCMS(ESI)[M+H]+:254.1。3-(Chloromethyl)-1-methylazetidine (1.2 g, 10 mmol, 1.0 eq) was dissolved in anhydrous ethanol (10 mL), followed by the addition of 2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-ol (1.7 g, 10.0 mmol, 1.0 eq) and potassium carbonate (4.1 g, 30.0 mmol, 3.0 eq). The mixture was allowed to react at 80°C for 5 hours. Upon completion, the reaction was purified by reverse-phase column chromatography to afford 6-[(1-methylazetidin-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine as a brown oil (560 mg, 22.1% yield). LCMS (ESI) [M+H]+: 254.1.

步骤3：2'-氯-5'-甲氧基-6-甲基-N-{6-[(1-甲基氮杂环丁烷-3-基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺的合成Step 3: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[(1-methylazetidin-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide

将6-[(1-甲基氮杂环丁烷-3-基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-胺(230mg,0.91mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(253mg,0.91mmol,1.0eq)溶于超干N,N-二甲基甲酰胺(5mL)中，加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(381mg,1.36mmol,1.5eq)和N-甲基咪唑(224mg,2.73mmol,3.0eq)，随后室温反应5小时。待反应完全，制备得白色固体2'-氯-5'-甲氧基-6-甲基-N-{6-[(1-甲基氮杂环丁烷-3-基)甲氧基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(34mg,收率7.3％)。LCMS(ESI)[M+H]+:514.1。6-[(1-Methylazetidin-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (230 mg, 0.91 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (253 mg, 0.91 mmol, 1.0 eq) were dissolved in ultra-dry N,N-dimethylformamide (5 mL), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (381 mg, 1.36 mmol, 1.5 eq) and N-methylimidazole (224 mg, 2.73 mmol, 3.0 eq) were added, followed by reaction at room temperature for 5 hours. After the reaction was complete, a white solid, 2'-chloro-5'-methoxy-6-methyl-N-{6-[(1-methylazetidin-3-yl)methoxy]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (34 mg, yield 7.3%) was obtained. LCMS (ESI) [M+H]+: 514.1.

实施例38：2'-氯-5'-甲氧基-6-甲基-N-{6-[5-(2,2,2-三氟乙基)氨基]吡啶-2-基}-4,5,6,7-四氢苯并[d]噻唑-2-基-[4,4'-联吡啶]-3-甲酰胺(化合物63)的合成
Example 38: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[5-(2,2,2-trifluoroethyl)amino]pyridin-2-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide (Compound 63)

步骤1：6-溴-N-(2,2,2-三氟乙基)吡啶-3-胺的合成Step 1: Synthesis of 6-bromo-N-(2,2,2-trifluoroethyl)pyridin-3-amine

将2-溴吡啶-5-硼酸(5.00g,24.9mmol,1.0eq)，2,2,2-三氟乙胺(7.39g,74.7mmol,3.0eq)，醋酸铜(6.76g,37.3mmol,1.5eq)，三乙胺(7.54g,74.7mmol,3.0eq)加入到乙腈(50mL)中，80℃反应2小时。反应结束后，硅藻土过滤除去不溶物，浓缩滤液，正相柱层析，得到黄色固体6-溴-N-(2,2,2-三氟乙基)吡啶-3-胺(427mg，收率6.7％)。LCMS(ESI)[M+H]+:254.9。2-Bromopyridine-5-boronic acid (5.00 g, 24.9 mmol, 1.0 eq), 2,2,2-trifluoroethylamine (7.39 g, 74.7 mmol, 3.0 eq), copper acetate (6.76 g, 37.3 mmol, 1.5 eq), and triethylamine (7.54 g, 74.7 mmol, 3.0 eq) were added to acetonitrile (50 mL) and reacted at 80°C for 2 hours. After completion of the reaction, the insoluble material was removed by filtration through Celite, and the filtrate was concentrated and purified by normal phase column chromatography to obtain 6-bromo-N-(2,2,2-trifluoroethyl)pyridin-3-amine (427 mg, 6.7% yield) as a yellow solid. LCMS (ESI) [M+H]+: 254.9.

步骤2：6-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)-N-(2,2,2-三氟乙基)吡啶-3-胺的合成Step 2: Synthesis of 6-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)pyridin-3-amine

将6-溴-N-(2,2,2-三氟乙基)吡啶-3-胺(427mg,1.67mmol,1.0eq)，4,4,5,5-四甲基-2-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)-1,3,2-二氧杂硼烷(489mg,1.84mmol,1.1eq)，1,1-双(二苯基膦)二荗铁二氯化钯(122mg,0.167mmol,0.1eq)和碳酸钾(577mg,4.18mmol,2.5eq)加入到1,4-二氧六环(10ml)和水(2ml)中。氮气氛围下80℃反应4小时。反应结束后，过滤，滤液加入乙酸乙酯(20mLx2)和水(15mL)萃取，浓缩有机相，正相柱层析，得到棕色油状物6-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)-N-(2,2,2-三氟乙基)吡啶-3-胺(328mg，收率62.3％)。LCMS(ESI)[M+H]+:315.1。6-Bromo-N-(2,2,2-trifluoroethyl)pyridin-3-amine (427 mg, 1.67 mmol, 1.0 eq), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (489 mg, 1.84 mmol, 1.1 eq), 1,1-bis(diphenylphosphino)diborane iron palladium dichloride (122 mg, 0.167 mmol, 0.1 eq), and potassium carbonate (577 mg, 4.18 mmol, 2.5 eq) were added to 1,4-dioxane (10 ml) and water (2 ml). The mixture was reacted at 80°C under a nitrogen atmosphere for 4 hours. After the reaction, the mixture was filtered, and the filtrate was extracted with ethyl acetate (20 mL x 2) and water (15 mL). The organic phase was concentrated and purified by normal phase column chromatography to obtain 6-(1,4-dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)pyridin-3-amine (328 mg, 62.3% yield) as a brown oil. LCMS (ESI) [M+H]+: 315.1.

步骤3：6-(1,4-二氧阿司匹林[4.5]癸-8-基)-N-(2,2,2-三氟乙基)吡啶-3-胺的合成Step 3: Synthesis of 6-(1,4-dioxyaspirino[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)pyridin-3-amine

将6-(1,4-二氧阿司匹林[4.5]癸-7-烯-8-基)-N-(2,2,2-三氟乙基)吡啶-3-胺(328mg,1.04mmol,1.0eq)和钯碳(66mg)加入到四氢呋喃(6ml)中，室温下搅拌16小时。反应结束后，硅藻土过滤，浓缩滤液，得到棕色油状物6-(1,4-二氧阿司匹林[4.5]癸-8-基)-N-(2,2,2-三氟乙基)吡啶-3-胺(290mg，收率87.9％)。LCMS(ESI)[M+H]+:317.1。6-(1,4-Dioxyaspirino[4.5]dec-7-en-8-yl)-N-(2,2,2-trifluoroethyl)pyridin-3-amine (328 mg, 1.04 mmol, 1.0 eq) and palladium on carbon (66 mg) were added to tetrahydrofuran (6 ml) and stirred at room temperature for 16 hours. After completion of the reaction, the mixture was filtered through celite and the filtrate was concentrated to obtain 6-(1,4-Dioxyaspirino[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)pyridin-3-amine (290 mg, 87.9% yield) as a brown oil. LCMS (ESI) [M+H]+: 317.1.

步骤4：4-[5-(2,2,2-三氟乙基)氨基吡啶-2-基]环己-1-酮的合成Step 4: Synthesis of 4-[5-(2,2,2-trifluoroethyl)aminopyridin-2-yl]cyclohexan-1-one

将6-(1,4-二氧阿司匹林[4.5]癸-8-基)-N-(2,2,2-三氟乙基)吡啶-3-胺(290mg,0.92mmol,1.0eq)和对甲苯磺酸一水合物(174mg,0.92mmol,1.0eq)加入到丙酮(9ml)和水(3ml)中，60℃反应1小时。反应结束后，加入乙酸乙酯(20mL)和水(15mL)萃取，浓缩有机相，正相柱分离纯化，得到棕色油状物4-[5-(2,2,2-三氟乙基)氨基吡啶-2-基]环己-1-酮(173mg，收率68.8％)。LCMS(ESI)[M+H]+:273.1。6-(1,4-Dioxyaspirin[4.5]dec-8-yl)-N-(2,2,2-trifluoroethyl)pyridin-3-amine (290 mg, 0.92 mmol, 1.0 eq) and p-toluenesulfonic acid monohydrate (174 mg, 0.92 mmol, 1.0 eq) were added to acetone (9 ml) and water (3 ml) and reacted at 60°C for 1 hour. After completion of the reaction, ethyl acetate (20 mL) and water (15 mL) were added for extraction. The organic phase was concentrated and purified by normal phase column chromatography to obtain 4-[5-(2,2,2-trifluoroethyl)aminopyridin-2-yl]cyclohexan-1-one (173 mg, 68.8% yield) as a brown oil. LCMS (ESI) [M+H]+: 273.1.

步骤5：6-{5-[(2,2,2-三氟乙基)氨基]吡啶-2-基}-4,5,6,7-四氢苯并[d]噻唑-2-胺的合成Step 5: Synthesis of 6-{5-[(2,2,2-trifluoroethyl)amino]pyridin-2-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine

将4-[5-(2,2,2-三氟乙基)氨基吡啶-2-基]环己-1-酮(173mg,0.63mmol,1.0eq)，单质碘(194mg,0.76mmol,1.2eq)和硫脲(96mg,1.26mmol,2.0eq)加入到乙醇(5ml)中，在80℃下搅拌16小时。反应结束后，加入碳酸氢钠饱和水溶液(15mL)和硫代硫酸钠饱和水溶液(15mL)，再加入乙酸乙酯(20mL)和水(15mL)萃取，浓缩有机相，得到棕色固体粗品6-{5-[(2,2,2-三氟乙基)氨基]吡啶-2-基}-4,5,6,7-四氢苯并[d]噻唑-2-胺(305mg)。LCMS(ESI)[M+H]+:329.1。4-[5-(2,2,2-trifluoroethyl)aminopyridin-2-yl]cyclohexan-1-one (173 mg, 0.63 mmol, 1.0 eq), iodine (194 mg, 0.76 mmol, 1.2 eq), and thiourea (96 mg, 1.26 mmol, 2.0 eq) were added to ethanol (5 mL) and stirred at 80°C for 16 hours. After completion of the reaction, saturated aqueous sodium bicarbonate (15 mL) and saturated aqueous sodium thiosulfate (15 mL) were added, followed by extraction with ethyl acetate (20 mL) and water (15 mL). The organic phase was concentrated to afford crude 6-{5-[(2,2,2-trifluoroethyl)amino]pyridin-2-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (305 mg) as a brown solid. LCMS (ESI) [M+H]+: 329.1.

步骤6：2'-氯-5'-甲氧基-6-甲基-N-{6-[5-(2,2,2-三氟乙基)氨基]吡啶-2-基}-4,5,6,7-四氢苯并[d]噻唑-2-基-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[5-(2,2,2-trifluoroethyl)amino]pyridin-2-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide

将6-{5-[(2,2,2-三氟乙基)氨基]吡啶-2-基}-4,5,6,7-四氢苯并[d]噻唑-2-胺(208mg,0.636mmol,1.0eq)，2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(176mg,0.636mmol,1.0eq)，N,N,N',N'-四甲基氯甲脒六氟磷酸盐(267mg,0.954mmol,1.5eq)和N-甲基咪唑(208mg,2.544mmol,4.0eq)加入到乙腈(10ml)中，室温反应1小时。反应结束后，浓缩反应液，高效液相制备色谱分离，得到白色固体2'-氯-5'-甲氧基-6-甲基-N-{6-[5-(2,2,2-三氟乙基)氨基]吡啶-2-基}-4,5,6,7-四氢苯并[d]噻唑-2-基-[4,4'-联吡啶]-3-甲酰胺(9.2mg，两步收率2.4％)。LCMS(ESI)[M+H]+:589.1381；1H NMR(400MHz,DMSO)δ12.40(s,1H),8.78(s,1H),8.17(s,1H),8.05(d,J＝2.5Hz,1H),7.51(s,1H),7.40(s,1H),7.10(s,2H),6.34(t,J＝6.9Hz,1H),3.95(m,J＝9.7,6.8Hz,2H),3.62(s,3H),3.05–2.96(m,1H),2.87(t,J＝7.1Hz,2H),2.70(d,J＝6.8Hz,2H),2.58(s,3H),2.11–1.86(m,2H).。6-{5-[(2,2,2-trifluoroethyl)amino]pyridin-2-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-amine (208 mg, 0.636 mmol, 1.0 eq), 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (176 mg, 0.636 mmol, 1.0 eq), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (267 mg, 0.954 mmol, 1.5 eq) and N-methylimidazole (208 mg, 2.544 mmol, 4.0 eq) were added to acetonitrile (10 ml) and reacted at room temperature for 1 hour. After the reaction, the reaction solution was concentrated and separated by high performance liquid chromatography to give a white solid 2'-chloro-5'-methoxy-6-methyl-N-{6-[5-(2,2,2-trifluoroethyl)amino]pyridin-2-yl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl-[4,4'-bipyridine]-3-carboxamide (9.2 mg, two-step yield 2.4%). LCMS(ESI)[M+H]+:589.1381;1H NMR(400MHz,DMSO)δ12.40(s,1H),8.78(s,1H ),8.17(s,1H),8.05(d,J＝2.5Hz,1H),7.51(s,1H),7.40(s,1H),7.10(s,2H),6. 34(t,J＝6.9Hz,1H),3.95(m,J＝9.7,6.8Hz,2H),3.62(s,3H),3.05–2.96(m,1H) ,2.87(t,J＝7.1Hz,2H),2.70(d,J＝6.8Hz,2H),2.58(s,3H),2.11–1.86(m,2H).

实施例39：2'-氯-5'-甲氧基-6-甲基-N-{6-[5-(N-甲基氨磺酰基)吡啶-2-基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(化合物64)的合成
Example 39: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[5-(N-methylsulfamoyl)pyridin-2-yl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (Compound 64)

步骤1：6-氯吡啶-3-磺酰氯的合成Step 1: Synthesis of 6-chloropyridine-3-sulfonyl chloride

(a)0℃下，将SOCl2(11mL)滴加至加入(70mL)中，随后向体系中加入CuCl(115mg,1.17mmol,0.03eq)，继续反应30分钟。(b)0℃下，将6-氯吡啶-3-胺(5g,38.89mmol,1.0eq)分批加入浓HCl(40mL)中，然后加入NaNO2(3g,43.48mol,1.1eq)的水(10mL)溶液，继续反应10分钟。0℃下，将溶液(b)缓慢加入溶液(a)中，继续反应2小时。过滤、浓缩，得白色固体6-氯吡啶-3-磺酰氯(6.6g,收率80.2％)。LCMS(ESI)[M+H]+:212.04。(a) SOCl₂ (11 mL) was added dropwise to a 70 mL solution at 0°C. CuCl (115 mg, 1.17 mmol, 0.03 eq) was then added to the system and the reaction continued for 30 minutes. (b) 6-Chloropyridin-3-amine (5 g, 38.89 mmol, 1.0 eq) was added portionwise to concentrated HCl (40 mL) at 0°C. A solution of NaNO₂ (3 g, 43.48 mol, 1.1 eq) in water (10 mL) was then added and the reaction continued for 10 minutes. Solution (b) was slowly added to solution (a) at 0°C and the reaction continued for 2 hours. Filtration and concentration afforded 6-chloropyridine-3-sulfonyl chloride (6.6 g, 80.2% yield) as a white solid. LCMS (ESI) [M+H]⁺: 212.04.

步骤2：6-氯-N-甲基吡啶-3-磺酰胺的合成Step 2: Synthesis of 6-chloro-N-methylpyridine-3-sulfonamide

将6-氯吡啶-3-磺酰氯(6g,28.29mol,1.0eq)，甲胺盐酸盐(2.3g,0.034mol,1.2eq)和三乙胺(8.6g,0.085mol,3.0eq)依次加至二氯甲烷(20ml)中，常温下反应2小时。向体系中加入H2O(100mL)和乙酸乙酯(100mlx3)萃取分液，浓缩有机相得棕色油状物6-氯-N-甲基吡啶-3-磺酰胺(2.5g,收率43.3％)。LCMS(ESI)[M+H]+:207.00。6-Chloropyridine-3-sulfonyl chloride (6 g, 28.29 mol, 1.0 eq), methylamine hydrochloride (2.3 g, 0.034 mol, 1.2 eq), and triethylamine (8.6 g, 0.085 mol, 3.0 eq) were added sequentially to dichloromethane (20 ml) and allowed to react at room temperature for 2 hours. H₂O (100 mL) and ethyl acetate (100 ml x 3) were added to the system for extraction. The organic phase was concentrated to afford 6-chloro-N-methylpyridine-3-sulfonamide (2.5 g, 43.3% yield) as a brown oil. LCMS (ESI) [M+H]+: 207.00.

步骤3：N-甲基-6-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)吡啶-3-磺酰胺的合成Step 3: Synthesis of N-methyl-6-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyridine-3-sulfonamide

将6-氯-N-甲基吡啶-3-磺酰胺(1.0g,4.85mmol,1.0eq)，4,4,5,5-四甲基-2-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)-1,3,2-二氧杂硼烷(1.55g,5.82mmol,1.2eq)，碳酸铯(4.72g,14.5mmol,3.0eq)和[1,1'-双(二苯基膦基)二茂铁]二氯化钯(177mg,0.24mmol,0.05eq)加至1,4-二氧六环(10ml)和水(2.5ml)溶液中，N2氛围下80℃反应3小时。过滤，浓缩，正相柱层析，浓缩得黄色固体N-甲基-6-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)吡啶-3-磺酰胺(990mg,收率65.8％)。LCMS(ESI)[M+H]+:311.1。6-Chloro-N-methylpyridine-3-sulfonamide (1.0 g, 4.85 mmol, 1.0 eq), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (1.55 g, 5.82 mmol, 1.2 eq), cesium carbonate (4.72 g, 14.5 mmol, 3.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (177 mg, 0.24 mmol, 0.05 eq) were added to a solution of 1,4-dioxane (10 ml) and water (2.5 ml) and reacted at 80 ° C under N 2 atmosphere for 3 hours. The mixture was filtered, concentrated, and purified by normal phase column chromatography to obtain a yellow solid N-methyl-6-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyridine-3-sulfonamide (990 mg, yield 65.8%). LCMS (ESI) [M+H]+: 311.1.

步骤4：N-甲基-6-(1,4-二氧杂螺[4.5]癸-8-基)吡啶-3-磺酰胺的合成Step 4: Synthesis of N-methyl-6-(1,4-dioxaspiro[4.5]dec-8-yl)pyridine-3-sulfonamide

将N-甲基-6-(1,4-二氧杂螺[4.5]癸-7-烯-8-基)吡啶-3-磺酰胺(980mg,3.16mmol,1.0eq)和钯/碳(67mg,0.63mmol,0.2eq)加至甲醇中(5mL)中，H2氛围下反应16h。过滤、浓缩得黄色粗品固体N-甲基-6-(1,4-二氧杂螺[4.5]癸-8-基)吡啶-3-磺酰胺(1.0g)，直接用于下一步反应。LCMS(ESI)[M+H]+:313.11。N-Methyl-6-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyridine-3-sulfonamide (980 mg, 3.16 mmol, 1.0 eq) and palladium/carbon (67 mg, 0.63 mmol, 0.2 eq) were added to methanol (5 mL) and reacted under an H₂ atmosphere for 16 h. Filtration and concentration afforded a crude yellow solid of N-methyl-6-(1,4-dioxaspiro[4.5]dec-8-yl)pyridine-3-sulfonamide (1.0 g), which was used directly in the next reaction. LCMS (ESI) [M+H]⁺: 313.11.

步骤5：N-甲基-6-(4-氧代环己基)吡啶-3-磺酰胺的合成Step 5: Synthesis of N-methyl-6-(4-oxocyclohexyl)pyridine-3-sulfonamide

将N-甲基-6-(1,4-二氧杂螺[4.5]癸-8-基)吡啶-3-磺酰胺(1.0g,3.2mmol,1.0eq)和甲苯磺酸一水合物(183mg,0.96mmol,0.3eq)加至丙酮/水(6mL/3mL)中，随后60℃反应2小时。加入饱和碳酸氢钠水溶液调节PH至中性，用乙酸乙酯(15mLx2)萃取分液，浓缩有机相得棕色粗品固体N-甲基-6-(4-氧代环己基)吡啶-3-磺酰胺(550mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:269.09。N-Methyl-6-(1,4-dioxaspiro[4.5]dec-8-yl)pyridine-3-sulfonamide (1.0 g, 3.2 mmol, 1.0 eq) and toluenesulfonic acid monohydrate (183 mg, 0.96 mmol, 0.3 eq) were added to acetone/water (6 mL/3 mL) and reacted at 60°C for 2 hours. The pH was adjusted to neutral by adding saturated aqueous sodium bicarbonate. The mixture was extracted with ethyl acetate (15 mL x 2). The organic phase was concentrated to obtain a brown crude solid of N-methyl-6-(4-oxocyclohexyl)pyridine-3-sulfonamide (550 mg), which was used directly in the next reaction. LCMS (ESI) [M+H]+: 269.09.

步骤6：6-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-N-甲基吡啶-3-磺酰胺的合成Step 6: Synthesis of 6-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-N-methylpyridine-3-sulfonamide

将N-甲基-6-(4-氧代环己基)吡啶-3-磺酰胺(200mg,0.75mmol,1.0eq)溶于乙醇(5ml)中，冰浴下加入碘(227mg,0.89mmol,1.2eq)和硫脲(113mg,1.49mmol,2.0eq)。80℃反应2小时，反应完全后，加入饱和硫代硫酸钠溶液淬灭反应，再加EA(25mLx2)萃取有机相，浓缩干燥得棕色粗品油状物6-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-N-甲基吡啶-3-磺酰胺(60mg)，直接用于下一步反应。LCMS(ESI)[M+H]+:325.07。N-Methyl-6-(4-oxocyclohexyl)pyridine-3-sulfonamide (200 mg, 0.75 mmol, 1.0 eq) was dissolved in ethanol (5 ml). Iodine (227 mg, 0.89 mmol, 1.2 eq) and thiourea (113 mg, 1.49 mmol, 2.0 eq) were added under ice-cooling. The mixture was allowed to react at 80°C for 2 hours. After completion, the reaction was quenched by addition of saturated sodium thiosulfate solution. The organic phase was extracted with EA (25 mL x 2) and concentrated to dryness to obtain a crude brown oil, 6-(2-amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-N-methylpyridine-3-sulfonamide (60 mg), which was used directly in the next reaction. LCMS (ESI) [M+H]+: 325.07.

步骤7：2'-氯-5'-甲氧基-6-甲基-N-{6-[5-(N-甲基氨磺酰基)吡啶-2-基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺的合成Step 7: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-[5-(N-methylsulfamoyl)pyridin-2-yl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide

将6-(2-氨基-4,5,6,7-四氢苯并[d]噻唑-6-基)-N-甲基吡啶-3-磺酰胺(60mg,0.18mmol,1.0eq)和2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(51mg,0.18mmol,1.0eq)溶于N,N-二甲基甲酰胺(3mL)中，随后向体系中加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(78mg,0.27mmol,1.5eq)和N-甲基咪唑(46mg,0.54mmol,3.0eq)，室温反应5小时。反应结束后，浓缩反应液，高效液相制备色谱分离，浓缩得白色固体2'-氯-5'-甲氧基-6-甲基-N-{6-[5-(N-甲基氨磺酰基)吡啶-2-基]-4,5,6,7-四氢苯并[d]噻唑-2-基}-[4,4'-联吡啶]-3-甲酰胺(11mg，收率10.4％)。LCMS(ESI)[M+H]+:585.1125；1H NMR(400MHz,DMSO-d6)δ8.87(d,J＝2.5Hz,1H),8.77(s,1H),8.17(s,1H),8.11(m,J＝8.2,2.4Hz,1H),7.65(d,J＝8.2Hz,2H),7.52(s,1H),7.41(s,1H),3.62(s,3H),3.32–3.25(m,1H),2.99(t,J＝7.9Hz,2H),2.74(s,2H),2.58(s,3H),2.46(s,3H),2.20–2.02(m,2H)。6-(2-Amino-4,5,6,7-tetrahydrobenzo[d]thiazol-6-yl)-N-methylpyridine-3-sulfonamide (60 mg, 0.18 mmol, 1.0 eq) and 2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (51 mg, 0.18 mmol, 1.0 eq) were dissolved in N,N-dimethylformamide (3 mL), and then N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (78 mg, 0.27 mmol, 1.5 eq) and N-methylimidazole (46 mg, 0.54 mmol, 3.0 eq) were added to the system and reacted at room temperature for 5 hours. After the reaction, the reaction solution was concentrated and separated by high performance liquid chromatography to give 2'-chloro-5'-methoxy-6-methyl-N-{6-[5-(N-methylsulfamoyl)pyridin-2-yl]-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl}-[4,4'-bipyridine]-3-carboxamide (11 mg, yield 10.4%) as a white solid. LCMS(ESI)[M+H]+:585.1125;1H NMR(400MHz,DMSO-d6)δ8.87(d,J＝2.5Hz,1H),8.77(s,1H),8.17(s,1H),8.11(m,J＝8.2,2.4Hz,1H),7.65(d,J＝8.2 Hz,2H),7.52(s,1H),7.41(s,1H),3.62(s,3H),3.32–3.25(m,1H),2.99(t ,J＝7.9Hz,2H),2.74(s,2H),2.58(s,3H),2.46(s,3H),2.20–2.02(m,2H).

实施例40：N-{6-氨基-6-[4-(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-基)-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(化合物70)的合成
Example 40: Synthesis of N-{6-amino-6-[4-(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (Compound 70)

步骤1：2-甲基-N-(1,4-二氧杂螺[4.5]癸烷-8-亚基)丙烷-2-磺酰胺的合成Step 1: Synthesis of 2-methyl-N-(1,4-dioxaspiro[4.5]decan-8-ylidene)propane-2-sulfonamide

将1,4-二氧阿司匹林[4.5]癸烷-8-酮(16.0g,102.45mmol,1.0eq)溶于四氢呋喃(160mL)中，再加入2-甲基丙烷-2-磺酰胺(12.42g,102.45mmol,1.0eq)，随后加入钛酸四乙酯(46.73g,204.89mmol,2.0eq)，50℃反应16小时。反应完成后，在冰浴条件下，将反应液倒入饱和的碳酸氢钠溶液中，随后加硅藻土过滤，滤液加入乙酸乙酯(150mLx2)萃取，浓缩有机层，正相柱分离纯化得到白色固体2-甲基-N-(1,4-二氧杂螺[4.5]癸烷-8-亚基)丙烷-2-磺酰胺(12.36g,收率46.55％)。1,4-Dioxaspirino[4.5]decan-8-one (16.0 g, 102.45 mmol, 1.0 eq) was dissolved in tetrahydrofuran (160 mL), and 2-methylpropane-2-sulfonamide (12.42 g, 102.45 mmol, 1.0 eq) was added, followed by tetraethyl titanate (46.73 g, 204.89 mmol, 2.0 eq). The reaction was allowed to react at 50°C for 16 hours. After completion, the reaction solution was poured into saturated sodium bicarbonate solution in an ice bath, filtered through celite, and the filtrate was extracted with ethyl acetate (150 mL x 2). The organic layer was concentrated and purified by normal phase column chromatography to obtain 2-methyl-N-(1,4-dioxaspiro[4.5]decan-8-ylidene)propane-2-sulfonamide (12.36 g, 46.55% yield) as a white solid.

LCMS(ESI)[M+H]+:260.13。LCMS (ESI) [M+H] +: 260.13.

步骤2：N-(8-(4-氯苯基)-1,4-二氧杂螺[4.5]癸烷-8-基)-2-甲基丙烷-2-磺酰胺的合成Step 2: Synthesis of N-(8-(4-chlorophenyl)-1,4-dioxaspiro[4.5]decan-8-yl)-2-methylpropane-2-sulfonamide

在冰浴条件下，将四氢呋喃(35mL)溶解的2-甲基-N-(1,4-二氧杂螺[4.5]癸烷-8-亚基)丙烷-2-磺酰胺(6.46g,24.93mmol,1.0eq)溶液缓慢滴加到(4-氯苯基)溴化镁(65mL,1.0M,2.6eq)中，常温反应2小时。反应完成后，在冰浴条件下，将反应液倒入饱和的氯化铵溶液中，再加入乙酸乙酯(150mLx2)萃取，浓缩有机层，正相柱分离纯化得到白色固体N-(8-(4-氯苯基)-1,4-二氧杂螺[4.5]癸烷-8-基)-2-甲基丙烷-2-磺酰胺(2.87g,收率31.03％)。LCMS(ESI)[M+H]+:394.13。A solution of 2-methyl-N-(1,4-dioxaspiro[4.5]decan-8-ylidene)propane-2-sulfonamide (6.46 g, 24.93 mmol, 1.0 eq) dissolved in tetrahydrofuran (35 mL) was slowly added dropwise to (4-chlorophenyl)magnesium bromide (65 mL, 1.0 M, 2.6 eq) in an ice bath. The reaction was allowed to react at room temperature for 2 hours. After completion, the reaction solution was poured into saturated ammonium chloride solution in an ice bath and extracted with ethyl acetate (150 mL x 2). The organic layer was concentrated and purified using a normal phase column to afford N-(8-(4-chlorophenyl)-1,4-dioxaspiro[4.5]decan-8-yl)-2-methylpropane-2-sulfonamide (2.87 g, 31.03% yield) as a white solid. LCMS (ESI) [M+H]+: 394.13.

步骤3：2-甲基-N-{8-[4-(2,2,2-三氟乙基)氨基]苯基}-1,4-二氧杂螺[4.5]癸烷-8-基)丙烷-2-磺酰胺的合成Step 3: Synthesis of 2-methyl-N-{8-[4-(2,2,2-trifluoroethyl)amino]phenyl}-1,4-dioxaspiro[4.5]decan-8-yl)propane-2-sulfonamide

将N-(8-(4-氯苯基)-1,4-二氧杂螺[4.5]癸烷-8-基)-2-甲基丙烷-2-磺酰胺(1.00g,2.7mmol,1.0eq)溶于1,4-二氧六环(10mL)中，随后加入叔丁醇钠(777mg,8.09mmol,3.0eq)和甲磺酸(2-二叔丁基膦基-3.6-二甲氧基-2',4',6'-三异丙基-1,1'-联苯基)(2-氨基-1,1'-联苯-2-基)钯(II)(230mg,0.27mmol,0.1eq)，氮气置换后滴加2,2,2-三氟乙醇-1-胺(1.34g,13.48mmol,5.0eq)，100℃反应1小时。反应完成后，浓缩反应液，正相柱分离纯化，得到白色固体2-甲基-N-{8-[4-(2,2,2-三氟乙基)氨基]苯基}-1,4-二氧杂螺[4.5]癸烷-8-基)丙烷-2-磺酰胺(850mg,收率72.65％)。LCMS(ESI)[M+H]+:457.19。N-(8-(4-chlorophenyl)-1,4-dioxaspiro[4.5]decan-8-yl)-2-methylpropane-2-sulfonamide (1.00 g, 2.7 mmol, 1.0 eq) was dissolved in 1,4-dioxane (10 mL), followed by the addition of sodium tert-butoxide (777 mg, 8.09 mmol, 3.0 eq) and methanesulfonic acid (2-di-tert-butylphosphino-3.6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) (230 mg, 0.27 mmol, 0.1 eq). After nitrogen substitution, 2,2,2-trifluoroethanol-1-amine (1.34 g, 13.48 mmol, 5.0 eq) was added dropwise, and the reaction was carried out at 100°C for 1 hour. After the reaction was completed, the reaction solution was concentrated and purified by normal phase column separation to obtain 2-methyl-N-{8-[4-(2,2,2-trifluoroethyl)amino]phenyl}-1,4-dioxaspiro[4.5]decan-8-yl)propane-2-sulfonamide (850 mg, 72.65% yield) as a white solid. LCMS (ESI) [M+H]+: 457.19.

步骤4：2-甲基-N-(4-氧代-1-{4-[(2,2,2-三氟乙基)氨基]苯基}环己基)丙烷-2-磺酰胺的合成将2-甲基-N-(8-(4-(2,2,2-三氟乙基)氨基)苯基)-1,4-二氧杂螺[4.5]癸烷-8-基)丙烷-2-磺酰胺(800mg,1.84mmol,1.0eq)溶于丙酮(9mL)中，随后加入对甲苯磺酸一水合物(34mg,0.18mmol,0.1eq)和水(3mL)，60℃反应3小时。反应完成后，加乙酸乙酯(50mL)和水(50mL)萃取反应液，收集有机层，干燥浓缩，正相柱分离纯化，得到棕色油状物2-甲基-N-(4-氧代-1-{4-[(2,2,2-三氟乙基)氨基]苯基}环己基)丙烷-2-磺酰胺粗品(320mg,收率44.51％)。LCMS(ESI)[M+H]+:413.16。Step 4: Synthesis of 2-methyl-N-(4-oxo-1-{4-[(2,2,2-trifluoroethyl)amino]phenyl}cyclohexyl)propane-2-sulfonamide 2-Methyl-N-(8-(4-(2,2,2-trifluoroethyl)amino)phenyl)-1,4-dioxaspiro[4.5]decan-8-yl)propane-2-sulfonamide (800 mg, 1.84 mmol, 1.0 eq) was dissolved in acetone (9 mL), followed by addition of p-toluenesulfonic acid monohydrate (34 mg, 0.18 mmol, 0.1 eq) and water (3 mL), and the reaction was carried out at 60 °C for 3 hours. After completion of the reaction, the reaction mixture was extracted with ethyl acetate (50 mL) and water (50 mL). The organic layer was collected, dried, concentrated, and purified by normal phase column chromatography to obtain crude 2-methyl-N-(4-oxo-1-{4-[(2,2,2-trifluoroethyl)amino]phenyl}cyclohexyl)propane-2-sulfonamide (320 mg, 44.51% yield) as a brown oil. LCMS (ESI) [M+H]+: 413.16.

步骤5：6-{4-[(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2,6-二胺的合成Step 5: Synthesis of 6-{4-[(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazole-2,6-diamine

将2-甲基-N-(4-氧代-1-(4-(2,2,2-三氟乙基)氨基)苯基)环己基)丙烷-2-磺酰胺粗品(100mg,0.256mmol,1.0eq)溶于无水乙醇(2mL)中，再加入硫脲(39mg,0.513mmol,2.0eq)和碘单质(98mg,0.385mmol,1.5eq)，80℃反应3小时。反应完成后，浓缩反应液，得到棕色固体6-{4-[(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2,6-二胺(130mg,收率100.00％)。LCMS(ESI)[M+H]+:326.09。Crude 2-methyl-N-(4-oxo-1-(4-(2,2,2-trifluoroethyl)amino)phenyl)cyclohexyl)propane-2-sulfonamide (100 mg, 0.256 mmol, 1.0 eq) was dissolved in anhydrous ethanol (2 mL). Thiourea (39 mg, 0.513 mmol, 2.0 eq) and iodine (98 mg, 0.385 mmol, 1.5 eq) were added and reacted at 80°C for 3 hours. After completion of the reaction, the reaction solution was concentrated to afford 6-{4-[(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazole-2,6-diamine (130 mg, 100.00% yield) as a brown solid. LCMS (ESI) [M+H]+: 326.09.

步骤6：N-{6-氨基-6-[4-(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-基)-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺的合成Step 6: Synthesis of N-{6-amino-6-[4-(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

将2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-羧酸(81mg,0.29mmol 1.0eq)和6-(4-((2,2,2-三氟乙基)氨基)苯基)-4,5,6,7-四氢苯并[d]噻唑-2,6-二胺(100mg,0.29mmol 1.0eq)溶于乙腈(5mL)中，再加入N-甲基咪唑(96mg,1.17mmol,4.0eq)，随后在冰浴条件下加入N,N,N’,N’-四甲基氯甲脒六氟磷酸盐(123mg,0.44mmol,1.5eq)，室温下反应16小时。反应结束后，制备分离纯化，得到黄色固体N-{6-氨基-6-[4-(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-基)-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(1.1mg,收率0.63％)。LCMS(ESI)[M+H]+:586.12。1H NMR(400MHz,DMSO)δ8.78(s,1H),8.17(s,1H),7.54(s,1H),7.42(s,1H),7.34(d,J＝8.5Hz,2H),6.77(s,1H),6.73(d,J＝8.4Hz,2H),6.39(t,J＝6.9Hz,1H),3.94(p,J＝9.4Hz,2H),3.60(s,3H),3.43(s,2H),3.42(s,2H),2.83(dd,J＝16.4,7.1Hz,4H),2.58(s,3H).2'-Chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (81 mg, 0.29 mmol, 1.0 eq) and 6-(4-((2,2,2-trifluoroethyl)amino)phenyl)-4,5,6,7-tetrahydrobenzo[d]thiazole-2,6-diamine (100 mg, 0.29 mmol, 1.0 eq) were dissolved in acetonitrile (5 mL). N-Methylimidazole (96 mg, 1.17 mmol, 4.0 eq) was added, followed by N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (123 mg, 0.44 mmol, 1.5 eq) in an ice bath. The mixture was reacted at room temperature for 16 hours. After completion of the reaction, preparative separation and purification were performed to obtain a yellow solid, N-{6-amino-6-[4-(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (1.1 mg, 0.63% yield). LCMS (ESI) [M+H]+: 586.12. 1H NMR (400MHz, DMSO) δ8.78(s,1H),8.17(s,1H),7.54(s,1H),7.42(s,1H),7.34(d,J＝8.5Hz,2H),6.77(s,1H),6.73(d,J＝8.4Hz,2H ),6.39(t,J＝6.9Hz,1H),3.94(p,J＝9.4Hz,2H),3.60(s,3H),3.43(s,2H),3.42(s,2H),2.83(dd,J＝16.4,7.1Hz,4H),2.58(s,3H).

实施例41：2'-氯-5'-甲氧基-6-甲基-N-{6-(甲氨基)-6-[4-(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(化合物71)的合成Example 41: Synthesis of 2'-chloro-5'-methoxy-6-methyl-N-{6-(methylamino)-6-[4-(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (Compound 71)

将N-{6-氨基-6-[4-(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-基)-2'-氯-5'-甲氧基-6-甲基-[4,4'-联吡啶]-3-甲酰胺(75mg,0.124mmol 1.0eq)，多聚甲醛(6mg,0.20mmol,1.6eq)和氰基硼氢化钠(13mg,0.21mmol,1.6eq)溶于甲醇中(4mL)中，室温下反应16小时。反应结束后，制备分离纯化，得到黄色固体2'-氯-5'-甲氧基-6-甲基-N-(6-(甲氨基)-6-{4-[(2,2,2-三氟乙基)氨基]苯基}-4,5,6,7-四氢苯并[d]噻唑-2-基)-[4,4'-联吡啶]-3-甲酰胺(3.3mg,收率4.3％)。LCMS(ESI)[M+H]+:617.16。1H NMR(400MHz,DMSO)δ8.87(d,J＝6.7Hz,1H),8.21(d,J＝1.9Hz,1H),7.65(d,J＝12.4Hz,1H),7.57(d,J＝2.2Hz,1H),7.27(d,J＝8.4Hz,2H),6.73(d,J＝8.4Hz,2H),3.93(s,3H),3.75(d,J＝16.2Hz,2H),3.60(s,3H),3.18(d,J＝16.1Hz,1H),2.77–2.57(m,5H),2.40(dt,J＝11.9,6.0Hz,1H),2.20(t,J＝5.3Hz,3H),2.10(d,J＝11.5Hz,1H).N-{6-amino-6-[4-(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-2'-chloro-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (75 mg, 0.124 mmol 1.0 eq), paraformaldehyde (6 mg, 0.20 mmol, 1.6 eq) and sodium cyanoborohydride (13 mg, 0.21 mmol, 1.6 eq) were dissolved in methanol (4 mL) and reacted at room temperature for 16 hours. After the reaction, preparative separation and purification were performed to obtain a yellow solid 2'-chloro-5'-methoxy-6-methyl-N-(6-(methylamino)-6-{4-[(2,2,2-trifluoroethyl)amino]phenyl}-4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)-[4,4'-bipyridine]-3-carboxamide (3.3 mg, yield 4.3%). LCMS (ESI) [M+H]+: 617.16 . NMR (400MHz, DMSO) δ8.87(d,J＝6.7Hz,1H),8.21(d,J＝1.9Hz,1H),7.65(d,J＝12.4 Hz,1H),7.57(d,J＝2.2Hz,1H),7.27(d,J＝8.4Hz,2H),6.73(d,J＝8.4Hz,2H),3.93( s,3H),3.75(d,J＝16.2Hz,2H),3.60(s,3H),3.18(d,J＝16.1Hz,1H),2.77–2.57(m ,5H),2.40(dt,J＝11.9,6.0Hz,1H),2.20(t,J＝5.3Hz,3H),2.10(d,J＝11.5Hz,1H).

通过与实施例中描述的类似的方法，分别制备得到下表1中所列的化合物，其中反应物、试剂的量、保护和脱保护、溶剂和反应条件有适当变化。化合物的表征数据汇总于下表1。The compounds listed in Table 1 below were prepared by methods similar to those described in the Examples, with appropriate variations in reactants, reagent amounts, protection and deprotection, solvents, and reaction conditions. Characterization data for the compounds are summarized in Table 1 below.

表1：部分化合物的结构和表征

Table 1: Structure and characterization of some compounds

实验例1：本发明的化合物对HCT116 BRCA2-/-细胞增殖作用的测定Experimental Example 1: Determination of the effect of the compounds of the present invention on the proliferation of HCT116 BRCA2 -/ - cells

实验目的：该测试例的目的是测试化合物对HCT116 BRCA2-/-细胞增殖作用。Experimental purpose: The purpose of this test example is to test the effect of compounds on the proliferation of HCT116 BRCA2 -/- cells.

背景原理：DNA聚合酶theta(Polθ)是具有同源重组缺陷的合成致死型靶点，在双链断裂(DSB)的DNA损伤应答途径中具有重要作用。当同源重组介导的修复受到损害时(HR缺陷)，如BRCA1或BRCA2突变，Polθ高度表达并引导DSB修复朝向alt-EJ，开启微同源介导的末端连接(MMEJ)的DNA修复过程。在HR缺陷的情况下，Polθ的抑制通过毒性RAD51中间体的积累和alt-EJ修复途径的抑制，导致细胞死亡。因此，通过检测化合物对HCT116 BRCA2-/-细胞增殖作用的抑制率，可用于Polθ蛋白抑制剂的筛选。Background and Principle: DNA polymerase theta (Polθ) is a synthetic lethal target with homologous recombination deficiency and plays an important role in the DNA damage response pathway for double-strand breaks (DSBs). When homologous recombination-mediated repair is impaired (HR deficiency), such as in BRCA1 or BRCA2 mutations, Polθ is highly expressed and directs DSB repair toward alt-EJ, initiating the DNA repair process of microhomology-mediated end joining (MMEJ). In the case of HR deficiency, inhibition of Polθ leads to cell death through the accumulation of toxic RAD51 intermediates and inhibition of the alt-EJ repair pathway. Therefore, by measuring the inhibitory effect of compounds on the proliferation of HCT116 BRCA2 -/- cells, it can be used to screen for Polθ protein inhibitors.

具体实验流程：Specific experimental process:

构建HCT116 BRCA2敲除细胞，筛选单克隆。将处于对数生长期的HCT116 BRCA2-/-和WT细胞，接种与96孔板，每孔90uL，1000个/孔,37℃培养箱静置过夜。第二天，加入10μl不同浓度的化合物，最高浓度为100uM，10倍稀释，9个梯度(DMSO终浓度为1％)，37℃培养箱孵育6天。第6天，吸去旧培养基，加入110ul培养基(培养基和CCK8比例为100:10)，37℃孵育1-4h。450nM处检测吸光值，通过GraphPad软件处理计算IC50，通过和阳性药物对比，筛选化合物。Construct HCT116 BRCA2 knockout cells and screen for single clones. HCT116 BRCA2 -/- and WT cells in logarithmic growth phase were seeded into 96-well plates, with 1000 cells/well (90 μL) per well, and incubated overnight at 37°C. The next day, 10 μL of compound was added at various concentrations, with a maximum concentration of 100 μM. Nine 10-fold dilutions were performed (final DMSO concentration was 1%), and the cells were incubated at 37°C for 6 days. On day 6, the old culture medium was aspirated and 110 μL of culture medium (culture medium to CCK8 at a ratio of 100:10) was added. The cells were incubated at 37°C for 1-4 hours. The absorbance was measured at 450 nM, and the IC50 value was calculated using GraphPad software. Compounds were screened by comparing the results with those of active drugs.

本发明所采用的阳性对照化合物及其制备方法：ART558见中国专利申请中CN201980101280.6的化合物24，RP25见PCT申请WO2023050007A1中的化合物25，化合物A见WO2023233295中化合物4，各个对照化合物结构式分别如下：The positive control compounds and preparation methods used in the present invention are as follows: ART558 is compound 24 in Chinese patent application CN201980101280.6, RP25 is compound 25 in PCT application WO2023050007A1, and compound A is compound 4 in WO2023233295. The structural formulas of the control compounds are as follows:

ART558：对照化合物A： ART558: Control compound A:

表2：对HCT116 BRCA2-/-细胞增殖作用的测定结果
Table 2: Results of the assay on the proliferation of HCT116 BRCA2-/- cells

结果显示本发明化合物能抑制HCT116 BRCA2-/-细胞的增殖，IC50值达到nM级，部分化合物的活性令人惊讶的显著优于阳性对照，这一强抑制作用对与Polθ抑制有关的病症或疾病的治疗具有重要的治疗意义；而对HCT116 WT细胞抑制作用弱，具有优异的选择性。The results showed that the compounds of the present invention can inhibit the proliferation of HCT116 BRCA2-/- cells, with IC50 values reaching the nM level. The activity of some compounds is surprisingly significantly better than that of the positive control. This strong inhibitory effect has important therapeutic significance for the treatment of symptoms or diseases related to Polθ inhibition; while the inhibitory effect on HCT116 WT cells is weak, with excellent selectivity.

实验例2：本发明的化合物对DLD1 BRCA2-/--细胞增殖作用的测定Experimental Example 2: Determination of the effect of the compounds of the present invention on the proliferation of DLD1 BRCA2-/- cells

实验目的：该测试例的目的是测试化合物对DLD1 BRCA2-/-细胞增殖作用。Experimental purpose: The purpose of this test case is to test the effect of compounds on the proliferation of DLD1 BRCA2 -/- cells.

具体实验流程：Specific experimental process:

构建DLD1 BRCA2敲除细胞，筛选单克隆。将处于对数生长期的DLD1 BRCA2-/-和WT细胞，接种与96孔板，每孔90uL，1000个/孔,37℃培养箱静置过夜。第二天，加入10μl不同浓度的化合物，最高浓度为100uM，10倍稀释，9个梯度(DMSO终浓度为1％)，37℃培养箱孵育6天。第6天，吸去旧培养基，加入110ul培养基(培养基和CCK8比例为100:10)，37℃孵育1-4h。450nM处检测吸光值，通过GraphPad软件处理计算IC50，通过和阳性药物对比，筛选化合物。DLD1 BRCA2 knockout cells were constructed and single clones were screened. DLD1 BRCA2 -/- and WT cells in logarithmic growth phase were seeded into 96-well plates, with 1000 cells/well (90 μL) per well, and incubated overnight at 37°C. The next day, 10 μL of compound was added at various concentrations, with a maximum concentration of 100 μM. Nine 10-fold dilutions were performed (final DMSO concentration was 1%), and the cells were incubated at 37°C for 6 days. On the 6th day, the old culture medium was aspirated and 110 μL of culture medium (culture medium to CCK8 ratio of 100:10) was added. The cells were incubated at 37°C for 1-4 hours. The absorbance was measured at 450 nM, and the IC50 was calculated using GraphPad software. Compounds were screened by comparing with active drugs.

表3：对DLD1 BRCA2-/-细胞增殖作用的测定结果
Table 3: Results of the assay on the proliferation of DLD1 BRCA2-/- cells

结果显示本发明化合物能抑制DLD1 BRCA2-/-细胞的增殖，IC50值达到nM级，这一强抑制作用对与Polθ抑制有关的病症或疾病的治疗具有重要的治疗意义。The results showed that the compounds of the present invention can inhibit the proliferation of DLD1 BRCA2-/- cells with an IC50 value reaching nM level. This strong inhibitory effect has important therapeutic significance for the treatment of symptoms or diseases related to Polθ inhibition.

实验例3：本发明的化合物对Polθ蛋白聚合酶功能作用的测试Experimental Example 3: Testing of the Effects of the Compounds of the Present Invention on the Function of Polθ Protein Polymerase

实验目的：该测试例的目的是测试化合物对POLθ蛋白聚合酶功能的作用。Experimental Purpose: The purpose of this test case is to test the effect of the compound on the function of Polθ protein polymerase.

背景原理：Polθ是一个包含C-末端A家族DNA聚合酶和N末端超家族2(SF2)型DNA解旋酶，中间由长且保守程度较低的未知功能中心域隔开。ART558结合于Polθ的聚合酶催化结构域内的变构位点，并以剂量依赖性方式抑制MMEJ。因此，通过检测化合物对POLθ蛋白聚合酶功能的抑制作用，可用于Polθ蛋白抑制剂的筛选。Principle: Polθ consists of a C-terminal family A DNA polymerase and an N-terminal superfamily 2 (SF2) DNA helicase, separated by a long, poorly conserved central domain of unknown function. ART558 binds to an allosteric site within the Polθ polymerase catalytic domain and inhibits MMEJ in a dose-dependent manner. Therefore, compounds that inhibit Polθ polymerase function can be used to screen for Polθ inhibitors.

具体实验流程：Specific experimental process:

POLθ-polymerase domain蛋白表达：将POLθ-polymerase domain序列(residues 1819-2590)克隆到具有n端(His)6x标记和烟草腐蚀病毒(TEV)蛋白酶裂解位点的pET24N载体上。将构建的载体转化到大肠杆菌Rosetta(DE3)中，摇菌到OD为0.6时，加入1mM IPTG，12℃培养16h。收集菌体，进行超声破碎，离心取上清，通过Ni-NTA纯化蛋白，透析后测定蛋白浓度和纯度。POLθ-polymerase domain protein expression: The POLθ-polymerase domain sequence (residues 1819-2590) was cloned into the pET24N vector containing an N-terminal (His) 6x tag and a tobacco etch virus (TEV) protease cleavage site. The constructed vector was transformed into Escherichia coli Rosetta (DE3) and incubated at an OD of 0.6. 1 mM IPTG was added and cultured at 12°C for 16 h. The cells were harvested, ultrasonically disrupted, and the supernatant was centrifuged. The protein was purified by Ni-NTA and dialyzed for determination of protein concentration and purity.

将纯化的POLθ-polymerase domain蛋白加入待测缓冲液(20mM Tris,pH 7.80,50mM KC1,10mM MgCl,1mM DTT,0.01％ BSA,0.01％ Tween20)中,蛋白浓度为4nM，加入待测化合物(11点稀释系列待测化合物)。上述酶和测试复合抑制剂混合物在室温下培养了15分钟。将dNTP和引物(引物：5-GCG GCT GTC ATA AG-3。模板：5-GCT ACA TTG ACA ATG GCA TCA AAT CTC AGA TTG CGT CTT ATG ACA GCC GCG-3。引物：模板＝1：1.1)加入所有测试孔，室温避光孵育60min。通过每孔加入16uL的含0.5％ PicoGreen染料的TE反应终止液终止反应，避光孵育90min。酶标仪在485/520nm读取荧光，通过GraphPad软件处理计算IC50，通过和阳性药物RP25对比，筛选化合物。Purified POLθ-polymerase domain protein was added to assay buffer (20 mM Tris, pH 7.80, 50 mM KCl, 10 mM MgCl, 1 mM DTT, 0.01% BSA, 0.01% Tween 20) at a protein concentration of 4 nM. Test compounds (an 11-point dilution series) were then added. The enzyme and test compound inhibitor mixture were incubated at room temperature for 15 minutes. dNTPs and primers (primer: 5-GCG GCT GTC ATA AG-3; template: 5-GCT ACA TTG ACA ATG GCA TCA AAT CTC AGA TTG CGT CTT ATG ACA GCC GCG-3; primer: template = 1:1.1) were added to all assay wells and incubated at room temperature in the dark for 60 minutes. The reaction was terminated by adding 16 μL of TE stop buffer containing 0.5% PicoGreen dye to each well and incubated in the dark for 90 min. Fluorescence was read at 485/520 nm using a microplate reader. IC50 values were calculated using GraphPad software and compared to the positive drug RP25 for compound screening.

表4：对Polθ蛋白聚合酶功能作用的测试结果
Table 4: Test results on the function of Polθ protein polymerase

结果显示本发明化合物对Polθ-polymerase domain蛋白的聚合酶功能具有非常强的抑制作用，IC50值均达到nM级，部分化合物的抑制作用相当于或者优于阳性对照药。这一强抑制作用对与Polθ抑制有关的病症或疾病的治疗具有重要的治疗意义。The results showed that the compounds of the present invention have very strong inhibitory effects on the polymerase function of the Polθ-polymerase domain protein, with IC50 values reaching the nanomolar range. Some compounds have inhibitory effects comparable to or superior to the positive control drug. This strong inhibitory effect has important therapeutic implications for the treatment of conditions or diseases associated with Polθ inhibition.

实验例4：本发明的化合物在小鼠体内药代动力学测试Experimental Example 4: Pharmacokinetics of the compounds of the present invention in mice

实验目的：通过灌胃(PO)向ICR雌性小鼠给予本发明化合物，考察药代动力学特点Experimental purpose: To administer the compound of the present invention to ICR female mice by oral gavage (PO) to investigate its pharmacokinetic characteristics

具体实验流程：Specific experimental process:

将本发明化合物和对照化合物A(制备方法参考专利申请WO2023233295中化合物4)进行灌胃(50mg/kg)给药，PO溶媒：40％ PEG300+15％吐温80+45％水，给药当天配制。给药前0h和给药后0.25h，0.5h，1h，2h，4h，7h，24h(灌胃)，经颈静脉采血，每个样品采集约0.02mL，K2-EDTA抗凝，采集后放置冰上。血浆样品处理：血液样本采集后置于冰上，并于1小时之内离心分离血浆(离心条件：4000g，5分钟，2-8℃)。血浆样本在分析前存放时则放于-80℃冰箱内。通过不同时间点的血药浓度数据，运用Phoenix WinNonlin7.0计算药代动力学参数。实验结果见下表4：The compound of the present invention and control compound A (the preparation method refers to compound 4 in patent application WO2023233295) were administered by gavage (50 mg/kg), PO solvent: 40% PEG300 + 15% Tween 80 + 45% water, prepared on the day of administration. Blood was collected from the jugular vein 0h before administration and 0.25h, 0.5h, 1h, 2h, 4h, 7h, 24h after administration (gavage), and about 0.02mL of each sample was collected. K2-EDTA was used for anticoagulation and the samples were placed on ice after collection. Plasma sample processing: After blood collection, the blood samples were placed on ice and centrifuged within 1 hour to separate the plasma (centrifugation conditions: 4000g, 5 minutes, 2-8°C). Plasma samples were stored in a -80°C refrigerator before analysis. Phoenix WinNonlin7.0 was used to calculate the pharmacokinetic parameters based on the blood drug concentration data at different time points. The experimental results are shown in Table 4 below:

表5：在ICR雌性小鼠的体内药代动力学测试结果
Table 5: In vivo pharmacokinetic test results in ICR female mice

本发明化合物在小鼠体内药代动力学数据显示，与对照化合物A相比，所测试代表性化合物在小鼠体内口服暴露量出乎意料的良好。The pharmacokinetic data of the compounds of the present invention in mice showed that the oral exposure of the tested representative compounds in mice was unexpectedly good compared with the control compound A.

Claims (19)

如式(I)所示的化合物或其立体异构体、互变异构体、药学上可接受的盐：
The compound represented by formula (I) or its stereoisomers, tautomers, and pharmaceutically acceptable salts:
其中，in, R1、R2、R3、R4、R5、R6和R7分别独立地选自H、氘、卤素、氨基、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基；R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 are each independently selected from H, deuterium, halogen, amino, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, C 1 -C 6 alkoxy; R8选自H、羟基、C1-C6烷氧基、NRaRb；R 8 is selected from H, hydroxy, C 1 -C 6 alkoxy, NR a R b ; R9为苯基、杂芳基、杂环基、-O(CH2)r-环烷基、-O(CH2)r-杂环烷基、-O(CH2)r-杂芳基，所述苯基、杂芳基、杂环基、-O(CH2)r-环烷基、-O(CH2)r-杂环烷基、-O(CH2)r-杂芳基可任选被1个或多个Rn取代；R 9 is phenyl, heteroaryl, heterocyclyl, -O(CH 2 ) r -cycloalkyl, -O(CH 2 ) r -heterocycloalkyl, -O(CH 2 ) r -heteroaryl, and the phenyl, heteroaryl, heterocyclyl, -O(CH 2 ) r -cycloalkyl, -O(CH 2 ) r -heterocycloalkyl, -O(CH 2 ) r -heteroaryl may be optionally substituted with one or more R n ; Rn选自H、卤素、氰基、NRaRb、羟基、C1-C6烷基、C3-C6环烷基、C1-C6羟基烷基、C1-C6烷氨基、C1-C6卤代烷氧基、-(CH2)r-O-C1-C6烷基、-C(O)NRaRb、-S(O)2NRaRb、-NH(CH2)rC1-C6卤代烷基、-NH(CH2)rOH、-(CH2)rOC1-C6卤代烷基、C1-C6卤代烷基；R n is selected from H, halogen, cyano, NR a R b , hydroxy, C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, C 1 -C 6 hydroxyalkyl, C 1 -C 6 alkylamino, C 1 -C 6 haloalkoxy, -(CH 2 ) r -OC 1 -C 6 alkyl, -C(O)NR a R b , -S(O) 2 NR a R b , -NH(CH 2 ) r C 1 -C 6 haloalkyl, -NH(CH 2 ) r OH, -(CH 2 ) r OC 1 -C 6 haloalkyl, C 1 -C 6 haloalkyl; Ra、Rb各自独立地选自不存在、氢、C1-C6烷基；R a and R b are each independently selected from absent, hydrogen, and C 1 -C 6 alkyl; r为0至3的整数。r is an integer from 0 to 3. 根据权利要求1所述的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中所述R9选自可任选被1个或多个Rn取代的苯基、含1至3个选自N、O、S的5-6元杂芳基、含1至3个选自N、O、S的5-6杂环基、-O(CH2)r-C3-C6环烷基、-O(CH2)r-C3-C6杂环烷基、-O(CH2)r-5-6元杂芳基。The compound according to claim 1 or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R 9 is selected from a phenyl group which may be optionally substituted by one or more R n , a 5-6 membered heteroaryl group containing one to three groups selected from N, O, and S, a 5-6 membered heterocyclic group containing one to three groups selected from N, O, and S, -O(CH 2 ) r -C 3 -C 6 cycloalkyl group, -O(CH 2 ) r -C 3 -C 6 heterocycloalkyl group, or -O(CH 2 ) r -5-6 membered heteroaryl group. 根据权利要求1-2任一项所述的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中所述R9选自
The compound according to any one of claims 1 to 2, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, wherein R 9 is selected from
根据权利要求1-3任一项所示的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中R1选自氢。The compound according to any one of claims 1 to 3 or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from hydrogen. 根据权利要求1-4任一项所示的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中R2选自甲基。The compound according to any one of claims 1 to 4 or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, wherein R 2 is selected from methyl. 根据权利要求1-5任一项所示的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中R3选自氢。The compound according to any one of claims 1 to 5 or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, wherein R 3 is selected from hydrogen. 根据权利要求1-6任一项所示的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中R4选自甲氧基。The compound according to any one of claims 1 to 6 or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, wherein R 4 is selected from methoxy. 根据权利要求1-7任一项所示的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中R5选自氢。The compound according to any one of claims 1 to 7 or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, wherein R 5 is selected from hydrogen. 根据权利要求1-8任一项所示的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中R7选自氢。The compound according to any one of claims 1 to 8, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, wherein R 7 is selected from hydrogen. 根据权利要求1-9任一项所示的化合物或其立体异构体、互变异构体、药学上可接受的盐，其具有结构式(I-1)：
The compound according to any one of claims 1 to 9 or its stereoisomers, tautomers, or pharmaceutically acceptable salts, which has the structural formula (I-1):
根据权利要求1-10任一项所示的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中R6选自卤素、氨基、C1-C6烷基、C1-C6卤代烷基，优选自氯、甲基、二氟甲基、氨基，更优选自氯。The compound according to any one of claims 1 to 10 or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R6 is selected from halogen, amino, C1 - C6 alkyl, C1 - C6 haloalkyl, preferably chlorine, methyl, difluoromethyl, amino, and more preferably chlorine. 根据权利要求1-11任一项所示的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中R8选自氢、羟基、甲氧基、氨基、氨基烷基，优选为氢、羟基、甲氧基、氨基、氨基甲基。The compound according to any one of claims 1 to 11 or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein R 8 is selected from hydrogen, hydroxy, methoxy, amino, and aminoalkyl, preferably hydrogen, hydroxy, methoxy, amino, and aminomethyl. 化合物或其立体异构体、互变异构体、药学上可接受的盐，其中所述化合物选自：


A compound or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the compound is selected from:


药物组合物，其包含根据权利要求1至13中任一项所述化合物及其立体异构体、互变异构体、或其药学上可接受的盐，以及一种或多种药学上可接受的载体或赋形剂。A pharmaceutical composition comprising the compound according to any one of claims 1 to 13, its stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers or excipients. 权利要求1至13任一项所定义的化合物或权利要求14所述的组合物在制备用于治疗Polθ过表达的疾病的药物中的用途，优选地所述疾病为癌症，更优选的所述癌症为实体瘤、乳腺癌、肺癌、膀胱癌、食道癌、前列腺癌、胰腺癌、宫颈癌。Use of the compound defined in any one of claims 1 to 13 or the composition according to claim 14 in the preparation of a medicament for treating a disease caused by overexpression of Polθ, preferably the disease is cancer, more preferably the cancer is a solid tumor, breast cancer, lung cancer, bladder cancer, esophageal cancer, prostate cancer, pancreatic cancer, or cervical cancer. 制备在患者中治疗同源重组(HR)缺陷癌症的药物的用途，包括向患者给药有效量的权利要求1至13任一项的化合物或权利要求14所述的组合物。Use of a method for preparing a medicament for treating homologous recombination (HR)-deficient cancer in a patient, comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 13 or a composition according to claim 14. 制备在患者中治疗癌症的药物的用途，包括向患者给药有效量的权利要求1至13任一项的化合物、其立体异构体、互变异构体、或其药学上可接受的盐或权利要求14所述的组合物，其中所述癌症的特征为BRCA基因表达的减少或不存在、BRCA基因的不存在、或BRCA蛋白功能的减少。Use of a medicament for treating cancer in a patient, comprising administering to the patient an effective amount of a compound according to any one of claims 1 to 13, a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, or the composition according to claim 14, wherein the cancer is characterized by reduced or absent BRCA gene expression, absent BRCA genes, or reduced BRCA protein function. 如式(II)所示的化合物或其立体异构体、互变异构体、药学上可接受的盐：
The compound represented by formula (II) or its stereoisomers, tautomers, and pharmaceutically acceptable salts:
其中，R8和R9定义如前面权利要求任一项所述。wherein R 8 and R 9 are as defined in any one of the preceding claims. 一种如下任一所示的化合物及其立体异构体、互变异构体或药学上可接受的盐，

A compound as shown below and its stereoisomers, tautomers or pharmaceutically acceptable salts,

PCT/CN2025/086748 2024-04-03 2025-04-02 POLθ INHIBITOR AND PHARMACEUTICAL COMPOSITION THEREOF AND USE Pending WO2025209499A1 (en)

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