WO2023109461A1 - 苯并噻吩类衍生物、及其制备方法和用途 - Google Patents

苯并噻吩类衍生物、及其制备方法和用途 Download PDF

Info

Publication number
WO2023109461A1
WO2023109461A1 PCT/CN2022/134097 CN2022134097W WO2023109461A1 WO 2023109461 A1 WO2023109461 A1 WO 2023109461A1 CN 2022134097 W CN2022134097 W CN 2022134097W WO 2023109461 A1 WO2023109461 A1 WO 2023109461A1
Authority
WO
WIPO (PCT)
Prior art keywords
preparation
compound
group
haloalkyl
difluoroethyl
Prior art date
Application number
PCT/CN2022/134097
Other languages
English (en)
French (fr)
Inventor
杨玉社
陆征宇
张丹
曹阳芷
Original Assignee
中国科学院上海药物研究所
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 中国科学院上海药物研究所 filed Critical 中国科学院上海药物研究所
Publication of WO2023109461A1 publication Critical patent/WO2023109461A1/zh

Links

Images

Classifications

    • 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
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
    • A61K31/381—Heterocyclic compounds having sulfur as a ring hetero atom having five-membered rings
    • 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
    • A61K31/397—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having four-membered rings, e.g. azetidine
    • 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
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/4025—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil not condensed and containing further heterocyclic rings, e.g. cromakalim
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A61K31/4162—1,2-Diazoles condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • 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
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/50—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
    • C07D333/52—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes
    • C07D333/62—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes 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 to carbon atoms of the hetero ring
    • C07D333/64—Oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems

Definitions

  • the invention belongs to the field of medicine, in particular to a class of benzothiophene derivatives, racemates, enantiomers, diastereomers and mixtures thereof, pharmaceutically acceptable salts and prodrugs thereof , the pharmaceutical composition containing it, its preparation method and its application in medicine.
  • the present invention relates to benzothiophene derivatives represented by general formula (I), their racemates, enantiomers, diastereoisomers and mixtures thereof, and pharmaceutically acceptable salts thereof , Prodrugs, pharmaceutical compositions containing them, use as selective estrogen receptor antagonists/degraders for the treatment of diseases, in particular estrogen receptor positive (ER + ) diseases.
  • Breast cancer is an estrogen-dependent tumor, and estrogen promotes the proliferation of breast cancer cells by interacting with estrogen receptors.
  • Estrogen receptors are steroid hormone receptors belonging to the nuclear receptor superfamily that mediate most of the known estrogenic effects.
  • ER ⁇ is distributed in breast, uterus, bone, aorta, liver and other tissues, while ER ⁇ is highly expressed in testis, ovarian granulosa cells, bone marrow and brain. Studies have shown that ER ⁇ is highly expressed in about 74% of breast cancer patients and is the main driver of breast cancer cell proliferation.
  • estrogen serves as an important signaling molecule involved in the development and homeostasis of multiple tissue types in both males and females. Estrogen binds to estrogen receptors and regulates basic functions such as gene expression, metabolism, cell growth and proliferation through cytoplasmic signaling pathways or activation of nuclear transcription.
  • endocrine therapy is the mainstay of treatment.
  • Traditional endocrine therapy drugs include aromatase inhibitors (AIs) such as letrozole and anastrozole (inhibit/decrease endogenous estrogen biosynthesis), and selective estrogen receptor modulators (SERMs) such as tamoxa Xifen, raloxifene (competitive binding to ER with estrogen).
  • AIs aromatase inhibitors
  • SERMs selective estrogen receptor modulators
  • SESDs Selective estrogen receptor degraders
  • fulvestrant is an analogue of estradiol 7 ⁇ -alkylation, which competes with estrogen for binding to ER, and attaches a hydrophobic part to the surface of the receptor, simulating the state of a partially unfolded protein, utilizing the unfolded state of the cytoplasmic protein
  • the unfolded protein response (UPR) system is used to degrade the target ER.
  • Fulvestrant is currently the only SERD approved by the FDA, with monthly doses of 500/250 mg intramuscularly for ER + locally advanced or metastatic postmenopausal breast cancer patients with recurrence or disease progression after endocrine therapy, or who have not previously received endocrine therapy Treatment of postmenopausal patients with HR + (progesterone receptor positive)/ HER2- (human epidermal growth factor receptor-2 negative) locally advanced/metastatic breast cancer.
  • HR + progesterone receptor positive
  • HER2- human epidermal growth factor receptor-2 negative
  • Patent applications for selective estrogen receptor degraders/antagonists with disclosed structures include LSZ-102 (US9322746), G1T48 (WO2018148576), GDC-0927 (WO2016097073), SAR439859 (CN108884079), AZD9833 (WO2019002442), GDC9545 (CN107108611).
  • the object of the present invention is to provide a class of novel structure, orally effective potent ER degradation agent/antagonist (bifunctional compound), which is used for diseases related to ER signaling pathway, especially breast cancer, ovarian cancer, colon cancer, prostate cancer , Treatment of endometrial cancer.
  • the first aspect of the present invention provides a compound represented by general formula (I), or its prodrug, enantiomer, diastereoisomer, racemate, or its pharmaceutically acceptable Salt,
  • R 1 is selected from: -COOH, -B(OH) 2 , C1-C6 alkyl, C1-C6 haloalkyl
  • R 2 is selected from: H, halogen, cyano, C1-C6 alkyl, C1- C6 haloalkyl
  • R 1 , R 2 form with the connected benzene ring
  • each R 3 and each R 4 are independently selected from: hydrogen, halogen, cyano, C1-C4 alkyl, C1-C4 Alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, -SO 2 NH 2 , -C(O)NH 2 ;
  • R 5 , R 6 are each independently selected from: hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, or R 5 , R 6 are connected with Carbon forms a C3-C6 carbocycle or a 3-6 membered heterocycle;
  • R is selected from: halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy;
  • o 0, 1, 2, 3 or 4;
  • Ring Y is Z 2 and Z 3 are each independently -CH- or -N-; ring A or ring B is the same or different, and each is independently C3-C6 cycloalkyl or 3-6 membered heterocycloalkyl; n is 0, 1, 2, 3 or 4;
  • n 0, 1, 2, 3 or 4;
  • R is halogen , cyano, hydroxyl, carboxyl or amino;
  • R9 is selected from halogen, cyano, amino, C1-C6 alkyl, C1-C6 haloalkyl.
  • R 1 is selected from: -COOH, -B(OH) 2 , C1-C4 alkyl, C1-C4 haloalkyl
  • R 2 is selected from: H, halogen, cyano, C1-C4 alkane Base, C1-C4 haloalkyl
  • R 1 and R 2 form with the connected benzene ring R9 is selected from halogen, cyano, amino, C1-C2 alkyl, C1-C2 haloalkyl.
  • R 5 and R 6 are each independently selected from: hydrogen, halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, or R 5.
  • R 6 and the connected carbon form a C3-C5 carbocycle or a 3-5 membered heterocycle;
  • R 7 is selected from: halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy.
  • R 1 is -COOH, -B(OH) 2 ;
  • R 2 is H;
  • R 1 and R 2 form with the connected benzene ring R9 is selected from hydrogen, fluorine, chlorine, cyano, amino, methyl, trifluoromethyl;
  • each R 3 is independently selected from: hydrogen, fluorine, chlorine, bromine, cyano, C1-C2 alkyl, C1-C2 alkoxy, C1-C2 haloalkyl, C1- C2 haloalkoxy;
  • each R is independently selected from: hydrogen, fluorine, chlorine, bromine;
  • R 5 and R 6 are each independently selected from: hydrogen, fluorine, chlorine, bromine, C1-C3 alkyl, C1-C2 haloalkyl, or R 5 and R 6 form a C3-C5 carbocycle with the connected carbon;
  • R is selected from: hydrogen, fluorine, chlorine, bromine, C1-C4 alkyl, C1-C2 haloalkyl;
  • o 0, 1, 2 or 3;
  • Ring Y is Z 2 and Z 3 are each independently -CH- or -N-; ring A or ring B is the same or different, and each independently represents C4 cycloalkyl or 4-membered heterocycloalkyl; n is 1, 2 or 3;
  • n 0, 1, 2 or 3;
  • R 8 is fluorine, chlorine, bromine or cyano.
  • R 1 is -COOH, -B(OH) 2 ;
  • R 2 is H; or R 1 and R 2 form
  • each R 3 is independently selected from: hydrogen, fluorine, chlorine, bromine, cyano, C1-C2 alkyl, C1-C2 alkoxy, C1-C2 haloalkyl, C1- C2 haloalkoxy;
  • each R is independently selected from: hydrogen, fluorine, chlorine, bromine;
  • R 5 and R 6 are each independently selected from: hydrogen, fluorine, chlorine, bromine, C1-C3 alkyl, C1-C2 haloalkyl, or R 5 and R 6 form a C3-C5 carbocyclic ring with the connected carbon;
  • R 7 Selected from: hydrogen, fluorine, chlorine, bromine, C1-C4 alkyl, C1-C2 haloalkyl;
  • Z 2 and Z 3 are each independently -CH- or -N-;
  • R 8 is fluoro, chloro, bromo or cyano.
  • ring Y is
  • -CR 5 R 6 R 7 is selected from: -CF 2 CH 3 , -CH(CF 3 )CH 3 , -CH 2 CF 3 , -CF(CH 3 ) 2 , -CF(CF 3 ) CH 3 , -C(CH 3 ) 2 CF 3 ,
  • the pharmaceutically acceptable salt is a salt formed by a compound and an organic acid or an inorganic acid, and the inorganic acid is selected from hydrochloric acid, phosphoric acid, sulfuric acid, hydrobromic acid, and nitric acid; the organic acid Selected from L-tartaric acid, fumaric acid, L-malic acid, D-malic acid, citric acid, L-pyroglutamic acid, acetic acid, p-toluenesulfonate, benzenesulfonic acid, methanesulfonic acid, benzoic acid, lactic acid , mandelic acid, maleic acid, oxalic acid, succinic acid.
  • the organic acid Selected from L-tartaric acid, fumaric acid, L-malic acid, D-malic acid, citric acid, L-pyroglutamic acid, acetic acid, p-toluenesulfonate, benzenesulfonic acid, methanesulfonic
  • the compound or a pharmaceutically acceptable salt thereof is selected from any compound prepared in Example 1-Example 54.
  • the second aspect of the present invention provides the preparation method of the compound described in the first aspect, wherein, when R 1 is -B(OH) 2 and R 2 is hydrogen, the preparation method comprises the following steps:
  • organoboron reagents selected from the group consisting of: boric acid, biboronic acid pinacol ester, pinacol borane, biboronic acid neopentyl glycol ester;
  • R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , Z 1 , Z 3 , m, n, o, p, and q are as described above;
  • X and W are respectively leaving groups, each independently selected from: halogen, trifluoromethanesulfonate group, mesylate group, p-toluenesulfonate group.
  • the preparation method comprises the following steps:
  • R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , Z 1 , Z 3 , m, n, o, p, and q are as described above;
  • W is a leaving group selected from the group consisting of halogen, trifluoromethanesulfonate, mesylate and p-toluenesulfonate.
  • the preparation method comprises the following steps:
  • R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , Z 2 , m, n, o, p, and q are as described above;
  • W, X are respectively leaving groups, each independently selected from: halogen, trifluoromethanesulfonate group, mesylate group, p-toluenesulfonate group;
  • PG is a protecting group, selected from: tert-butyl Oxycarbonyl, benzyloxycarbonyl, fluorenylmethyloxycarbonyl, allyloxycarbonyl, trifluoroacetyl, benzyl, p-methoxybenzyl, trityl.
  • preparation method comprises the following steps:
  • R 9 exists or does not exist
  • R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , Z 1 , Z 2 , m, n, o, p, q, and ring Y are as described above;
  • W, X are respectively leaving groups, each independently selected from: halogen, trifluoromethanesulfonate group, mesylate group, p-toluenesulfonate group;
  • PG is a protecting group, selected from: tert-butyl Oxycarbonyl, benzyloxycarbonyl, fluorenylmethyloxycarbonyl, allyloxycarbonyl, trifluoroacetyl, benzyl, p-methoxybenzyl, trityl.
  • the third aspect of the present invention provides a pharmaceutical composition, comprising: the compound described in the first aspect, or its prodrug, enantiomer, diastereoisomer, racemate, or its pharmaceutical an acceptable salt; and a pharmaceutically acceptable carrier.
  • the fourth aspect of the present invention provides the compound described in the first aspect, or its prodrug, enantiomer, diastereoisomer, racemate, or a pharmaceutically acceptable salt thereof or the third
  • the pharmaceutical composition described in the aspect is used for preparing medicines for treating and/or preventing diseases related to estrogen receptors.
  • the estrogen receptor-related diseases are selected from cancer, osteoporosis, neurodegenerative diseases, cardiovascular diseases, lupus erythematosus, endometriosis and obesity.
  • the cancer is selected from breast cancer, ovarian cancer, colon cancer, prostate cancer, and endometrial cancer.
  • the estrogen receptor-related diseases are selected from ER-positive breast cancer.
  • the estrogen receptor-related disease is selected from ER-positive brain metastatic breast cancer.
  • FIG 1 shows the inhibitory effect of different concentrations of compounds on the proliferation of MCF-7 Tam1 cells
  • Figure 2 shows the antagonism of different concentrations of compounds on ER ⁇ mutants Y537S and D538G
  • Figure 3 shows the effects of different concentrations of compounds on the degradation of MCF-7(Y537S) ER ⁇ protein.
  • Figure 4 is the result of uterine wet weight experiment, wherein A shows the average ratio of uterine weight to body weight in different experimental groups, *p ⁇ 0.05, **p ⁇ 0.01; B is the histological image of uterine cross-section in each experimental group, The height of the endometrial surface epithelium is indicated by a black line.
  • Fig. 5 is a graph showing the growth inhibition experiment results of the compound on the MCF-7 mouse subcutaneous tumor model, **p ⁇ 0.01.
  • the inventors of the present application have developed a compound with the structure shown in general formula I, which has dual functions of ER antagonism and ER degradation, and is a complete ER antagonist.
  • SERD can better cut off the ER signaling pathway and achieve better therapeutic effects on ER + related diseases.
  • the compounds of the invention have good brain tissue exposure. Therefore, the compound of formula (I) of the present invention is suitable for use in medicines for the treatment and/or prevention of estrogen receptor-related diseases or brain metastatic diseases. On this basis, the present invention has been accomplished.
  • Hydrogen in the compounds of the present invention include all isotopes thereof. Isotopes are understood to include those atoms having the same atomic number but different mass numbers. For example, hydrogen isotopes include tritium and deuterium, carbon isotopes include 13 C and 14 C, oxygen isotopes include 16 O and 18 O, etc.;
  • halogen means fluorine, chlorine, bromine or iodine
  • Alkyl means a saturated hydrocarbon group consisting only of the two elements C and H when any carbon atom loses a hydrogen atom, including straight and branched chain aliphatic hydrocarbons, non-limiting examples include methyl , ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl;
  • Alkylene refers to a straight or branched chain saturated aliphatic group, ie, a divalent hydrocarbon group, having the specified number of carbon atoms and linked to at least two other groups. Two groups attached to an alkylene group can be attached to the same or different atoms of the alkylene group.
  • a linear alkylene group can be a divalent group of -(CH2) n- , where n is 1, 2, 3, 4, 5 or 6.
  • Representative alkylene groups include, but are not limited to, methylene, ethylene, propylene, isopropylene, butylene, isobutylene, sec-butylene, pentylene, and hexylene.
  • Alkenyl means an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, such as ethenyl, 1-propenyl, 2-propenyl, etc.;
  • Cycloalkyl refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent.
  • Polycyclic cycloalkanes include spiro, fused and bridged cycloalkyls;
  • Heterocycloalkyl refers to a saturated non-aromatic cyclic group containing at least one (such as 1, 2, 3 or 4) ring heteroatoms (such as N, O or S), such as azetidinyl , piperidyl, tetrahydropyranyl, tetrahydropyridyl, pyrrolinyl, dihydropyridyl, dihydrofuranyl, dihydrothiophenyl, morpholinyl;
  • Spirocyclyl refers to a polycyclic group in which two monocyclic rings share one carbon atom, wherein the two rings can be carbocyclic, and can also contain 1 or more heteroatoms;
  • Alkoxy means -O-(alkyl) and -O-(cycloalkyl), wherein alkyl and cycloalkyl are as defined above;
  • substitution means that one or more hydrogen atoms in the group are independently replaced by the corresponding number of substituents
  • “Isomers” in the present invention refer to compounds having the same molecular formula but differing in properties or in the bond sequence of their atoms or in the arrangement of their atoms in space.
  • Stereoisomers are isomers that differ in the arrangement of their atoms in space.
  • Stereoisomers that are not mirror images of each other are diastereomers and stereoisomers that are non-superimposable mirror images of each other are enantiomers.
  • Chiral compounds can exist as single enantiomers or as mixtures thereof. A mixture comprising equal proportions of the enantiomers is called a "racemic mixture".
  • the “pharmaceutically acceptable salt” of the present invention refers to the salt of the compound of the present invention, which is safe and effective when used in mammals, and has proper biological activity.
  • the in vivo action of compounds of general formula (I) may partly be exerted by one or more metabolites formed in the human or animal body after administration of compounds of general formula (I).
  • the in vivo action of compounds of general formula (I) can also be exerted via the metabolism of precursor compounds "prodrugs".
  • the "prodrug” of the present invention refers to a compound that is converted into a compound of formula (I) due to reaction with enzymes, gastric acid, etc. under physiological conditions in an organism, that is, converted into A compound of the compound of the formula (I) and/or a compound converted into a compound of the formula (I) by a hydrolysis reaction such as gastric acid or the like.
  • Suitable pharmaceutically acceptable prodrugs of compounds of general formula (I) having a carboxyl group are, for example, their in vivo cleavable esters.
  • Suitable pharmaceutically acceptable esters for the carboxyl group include alkyl esters such as methyl, ethyl and tert-butyl, alkoxymethyl esters such as methoxymethyl; alkanoyloxymethyl esters; 3-phthalyl esters; cycloalkylcarbonyloxyalkyl esters such as cyclopentylcarbonyloxymethyl and 1-cyclohexylcarbonyloxyethyl; 2 -Oxo-1,3-dioxolenyl (dioxolenyl)methyl ester, such as 5-methyl-2-oxo-1,3-dioxolenyl-4-ylmethyl ester and alkoxycarbonyloxyalkyl esters such as methoxycarbonyloxymethyl ester and 1-methoxycarbonyloxye
  • Suitable pharmaceutically acceptable prodrugs of compounds of general formula (I) having a carboxyl group are, for example, amides cleavable in vivo, such as N-alkylamides and N,N-dialkylamides, such as N-methyl Amide, N-ethylamide, N-propylamide, N,N-dimethylamide, N-ethyl-N-methylamide or N,N-diethylamide.
  • the present invention provides general preparation methods for compounds represented by general formula (I).
  • Compounds of general formula (I) are synthesized according to the methods described below, as well as methods commonly used by synthetic organic chemists and combinations or variations of these methods.
  • the synthetic routes of the compounds in the present invention are not limited to the methods summarized below, individual compounds may need to adjust the operating conditions to meet the requirements of various functional groups. Various protecting groups known to those skilled in the art may be necessary. Additionally, the specific examples herein illustrate methods for synthesizing compounds of the invention.
  • R 1 is -B(OH) 2 ;
  • R 2 is hydrogen;
  • the racemate and enantiomer of the compound represented by the general formula (I) , diastereoisomers and mixtures thereof, pharmaceutically acceptable salts and prodrugs thereof can be prepared by route 1.
  • R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , Z 1 , Z 3 , m, n, o, p, q are as described above;
  • W is a leaving group, including but not limited to halogen, triflate, mesylate, p-toluenesulfonate, preferably X is triflate, W is bromine ;
  • A2 is obtained by selective Miyaura coupling of A1 in the presence of palladium catalyst, organoboron reagent and base.
  • the palladium catalysts described therein include, but are not limited to, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, tetrakis(triphenylphosphine)palladium, tris(dibenzylideneacetone)bis Palladium, bis(dibenzylideneacetone) palladium, bistriphenylphosphine palladium dichloride, bis(tri-tert-butylphosphine)palladium, bis(tricyclohexylphosphine)palladium, palladium acetate;
  • the organoboron reagent Including but not limited to boric acid, pinacol diborate, pinacol borane, neopentyl glycol diborate;
  • the base includes but not limited to potassium a
  • A3 is obtained by oxidation of A2.
  • Described oxidizing agent includes but not limited to sodium periodate;
  • A5 is obtained through the nucleophilic substitution reaction of A3 and A4 under basic conditions, and the base includes but not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dipotassium hydrogen phosphate, Cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, cesium fluoride, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, N,N-Diisopropylethylamine.
  • the base includes but not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dipotassium hydrogen phosphate, Cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, cesium fluoride, 1,8-diazabicyclo[5.4.0]undec-7
  • R 1 is -COOH; R 2 is hydrogen; the racemate of the compound shown in the general formula (I) , enantiomers, diastereomers and mixtures thereof, pharmaceutically acceptable salts and prodrugs thereof can be prepared by route 2.
  • R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , Z 1 , Z 3 , m, n, o, p, q are as described above;
  • W is a leaving group, including but not limited to halogen, trifluoromethanesulfonate, mesylate, p-toluenesulfonate, preferably W is bromine;
  • B3 is obtained by the nucleophilic addition and elimination reaction of B1 and B2 under alkaline conditions, and the base includes but not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dihydrogen phosphate Potassium, cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, cesium fluoride, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethyl Amine, N,N-Diisopropylethylamine;
  • B4 is obtained by reduction of B3 through sulfone group and aromatic bromination, wherein the reducing agent includes but not limited to diisobutylaluminum hydride, lithium aluminum hydride, trimethyl iodosilane, dihydrobis(2-methoxyethoxy)aluminum Sodium acid, sodium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, preferably diisobutylaluminum hydride; bromination reagents include but not limited to N-bromosuccinimide, liquid bromine, Tribromopyridinium salt, preferably N-bromosuccinimide;
  • the palladium catalyst used includes but not limited to [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride, tetrakis(triphenyl phosphine)palladium, tris(dibenzylideneacetone)dipalladium, bis(dibenzylideneacetone)palladium, bistriphenylphosphinepalladium dichloride, bis(tri-tert-butylphosphine)palladium, bis(tricyclo Hexylphosphine) palladium, palladium acetate
  • the base used includes but not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dipotassium hydrogen phosphate, cesium carbonate, sodium tert-butoxide, tert-butyl Potassium alkoxide, sodium methoxide,
  • B7 is obtained by removing the protective group of B6 under acidic conditions, wherein the acid includes but not limited to formic acid, acetic acid, methanesulfonic acid, hydrochloric acid, sulfuric acid, nitric acid, trifluoroacetic acid, trifluoromethanesulfonic acid, trichloride Boron, boron tribromide, aluminum trichloride;
  • the acid includes but not limited to formic acid, acetic acid, methanesulfonic acid, hydrochloric acid, sulfuric acid, nitric acid, trifluoroacetic acid, trifluoromethanesulfonic acid, trichloride Boron, boron tribromide, aluminum trichloride;
  • B9 is obtained by nucleophilic substitution reaction between B7 and B8 under basic conditions, and the base includes but not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dipotassium hydrogen phosphate, Cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, cesium fluoride, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, N,N-Diisopropylethylamine;
  • B10 is obtained by pinnick oxidation of B9.
  • the route is:
  • R 9 exists or does not exist, R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , Z 2 , m, n, o, p, q are as described above;
  • X, W are respectively leaving groups, including but not limited to halogen, trifluoromethanesulfonate group, mesylate group, p-toluenesulfonate group, preferably X is iodine, W is iodine;
  • PG is a protecting group Groups selected from: tert-butoxycarbonyl, benzyloxycarbonyl, fluorenylmethoxycarbonyl, allyloxycarbonyl, trifluoroacetyl, benzyl, p-methoxybenzyl, trityl;
  • the base includes but not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate , dipotassium hydrogen phosphate, cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, cesium fluoride, 1,8-diazabicyclo[5.4.0]undeca-7- ene, triethylamine, N,N-diisopropylethylamine;
  • C3 is obtained by reduction of C2, wherein the reducing agent includes but not limited to diisobutylaluminum hydride, lithium aluminum hydride, trimethyl iodosilane, dihydrobis(2-methoxyethoxy)sodium aluminate, preferably diisobutylaluminum hydride Butyl aluminum hydride;
  • the reducing agent includes but not limited to diisobutylaluminum hydride, lithium aluminum hydride, trimethyl iodosilane, dihydrobis(2-methoxyethoxy)sodium aluminate, preferably diisobutylaluminum hydride Butyl aluminum hydride;
  • C5 removes the benzyl protecting agent by catalytic hydrogenation reaction of C3, wherein the catalyst used includes but not limited to palladium/carbon, palladium hydroxide/carbon; then reacts with C4 under basic conditions for nucleophilic substitution, and the base used includes but Not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dipotassium hydrogen phosphate, cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, cesium fluoride , 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, N,N-diisopropylethylamine;
  • C6 is obtained by selective aromatic bromination of C5; bromination reagents include but not limited to N-bromosuccinimide, liquid bromine, tribromopyridinium salt, preferably N-bromosuccinimide;
  • the palladium catalysts used in Suzuki coupling include but are not limited to [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, tetrakis(triphenylphosphine)palladium, tris(dibenzylideneacetone) Dipalladium, bis(dibenzylideneacetone)palladium, bistriphenylphosphinepalladium dichloride, bis(tri-tert-butylphosphine)palladium, bis(tricyclohexylphosphine)palladium, palladium acetate
  • the base used includes but Not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dipotassium hydrogen phosphate, cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, cesium
  • C10 is obtained by removing the protecting group of C8 under acidic conditions, and further nucleophilic substitution reaction with C9 under basic conditions.
  • the acid includes but not limited to formic acid, acetic acid, methanesulfonic acid, hydrochloric acid, sulfuric acid, nitric acid, trifluoroacetic acid, trifluoromethanesulfonic acid, boron trichloride, boron tribromide, aluminum trichloride;
  • Bases include, but are not limited to, potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dipotassium hydrogen phosphate, cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, Cesium fluoride, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, N,N-diisopropylethylamine;
  • the route is:
  • R 9 exists or does not exist;
  • R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , Z 1 , Z 2 , m, n, o, p, q are as described above;
  • X, W are respectively leaving groups, including but not limited to halogen, trifluoromethanesulfonate group, mesylate group, p-toluenesulfonate group, preferably X is iodine, W is iodine;
  • PG is protection A group selected from: tert-butoxycarbonyl, benzyloxycarbonyl, fluorenylmethoxycarbonyl, allyloxycarbonyl, trifluoroacetyl, benzyl, p-methoxybenzyl, trityl;
  • the base includes but not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate , dipotassium hydrogen phosphate, cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, cesium fluoride, 1,8-diazabicyclo[5.4.0]undeca-7- ene, triethylamine, N,N-diisopropylethylamine;
  • D3 is obtained by reduction of D2, wherein the reducing agent includes but not limited to diisobutylaluminum hydride, lithium aluminum hydride, trimethyl iodosilane, dihydrobis(2-methoxyethoxy)sodium aluminate, preferably diisobutylaluminum hydride Butyl aluminum hydride;
  • the reducing agent includes but not limited to diisobutylaluminum hydride, lithium aluminum hydride, trimethyl iodosilane, dihydrobis(2-methoxyethoxy)sodium aluminate, preferably diisobutylaluminum hydride Butyl aluminum hydride;
  • the palladium catalysts used for coupling include but are not limited to [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, tetrakis(triphenylphosphine)palladium, tris(dibenzylideneacetone)bis Palladium, bis(dibenzylideneacetone)palladium, bistriphenylphosphinepalladium dichloride, bis(tri-tert-butylphosphine)palladium, bis(tricyclohexylphosphine)palladium, palladium acetate, chloro(2-bicyclo Hexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2-aminoethylphenyl)]palladium ( II), methanesulfonic acid (2-
  • D6 is obtained by selective aromatic bromination of D5; bromination reagents include but not limited to N-bromosuccinimide, liquid bromine, tribromopyridinium salt, preferably N-bromosuccinimide;
  • D8 was obtained by Suzuki coupling of D6 and D7.
  • the palladium catalysts used in Suzuki coupling include but are not limited to [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, tetrakis(triphenylphosphine)palladium, tris(dibenzylideneacetone) Dipalladium, bis(dibenzylideneacetone)palladium, bistriphenylphosphinepalladium dichloride, bis(tri-tert-butylphosphine)palladium, bis(tricyclohexylphosphine)palladium, palladium acetate
  • the base used includes but Not limited to potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dipotassium hydrogen phosphate, cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluor
  • D10 is obtained by removing the protecting group of D8 under acidic conditions, and then undergoes nucleophilic substitution reaction with D9 under basic conditions.
  • the acid includes but not limited to formic acid, acetic acid, methanesulfonic acid, hydrochloric acid, sulfuric acid, nitric acid, trifluoroacetic acid, trifluoromethanesulfonic acid, boron trichloride, boron tribromide, aluminum trichloride;
  • Bases include, but are not limited to, potassium acetate, potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, potassium phosphate, dipotassium hydrogen phosphate, cesium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium methoxide, potassium fluoride, Cesium fluoride, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, N,N-diisopropylethyl
  • the compound of general formula (I) is dissolved in a suitable solvent, and 1 equivalent of acid is dissolved in the same solvent or water. Then the acid solution was added dropwise to the solution of the compound of general formula (I), stirred for 30 minutes, and freeze-dried. Suspend the freeze-dried powder in methyl tert-butyl ether or n-hexane, beat for 30 minutes, filter, and dry in vacuum to obtain the corresponding salt.
  • organic acid is organic acid or inorganic acid
  • described inorganic acid includes but not limited to hydrochloric acid, phosphoric acid, sulfuric acid, hydrobromic acid, nitric acid
  • Organic acid includes but not limited to L-tartaric acid, fumaric acid, L-malic acid, D-malic acid, citric acid, L-pyroglutamic acid, acetic acid, p-toluenesulfonate, benzenesulfonic acid, methanesulfonic acid, benzoic acid, lactic acid, mandelic acid, maleic acid, oxalic acid, succinic acid.
  • the solvents mentioned therein include but not limited to acetonitrile, acetone, ethyl acetate, dichloromethane, tetrahydrofuran, methanol, ethanol, isopropanol, 1,4-dioxane, chloroform, ether.
  • compositions and pharmaceutical dosage forms containing compounds of the invention comprising:
  • the present invention is also a pharmaceutical composition, which contains the compound of general formula (I) and its racemate, enantiomer, diastereoisomer and mixture thereof, its pharmaceutically acceptable salt, pro
  • the body drug is mixed with a pharmaceutically acceptable carrier, adjuvant or vehicle.
  • the pharmaceutical composition of the present invention can be formulated into dosage forms suitable for oral, parenteral (including subcutaneous, intramuscular, cortical and intravenous), bronchial or nasal administration as required.
  • parenteral including subcutaneous, intramuscular, cortical and intravenous
  • bronchial or nasal administration as required.
  • the pharmaceutical composition of the present invention is configured into a dosage form suitable for oral administration.
  • compositions may also contain various excipients, for example, preservatives, wetting agents, emulsifying agents and dispersing agents.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.
  • the active compound is mixed with at least one inert excipient (or carrier), which may also include: (a) filler or mixing agent; (b) binder; (c) humectant; ( d) disintegrants; (e) solution retarders; (f) absorption enhancers; (g) wetting agents; (h) adsorbents; (i) lubricants;
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, dispersions, syrups or elixirs.
  • Liquid dosage forms may contain, in addition to the active compound, inert diluents (eg, parallel solvents), solubilizers, and emulsifiers commonly used in the art.
  • the compound of the present invention has dual functions of ER antagonism and ER degradation, and is a complete ER antagonist. In theory, it can better cut off the ER signaling pathway than SERD with partial agonistic activity, and achieve better therapeutic effects on ER + related diseases. Therefore, the compound of formula (I) of the present invention is suitable for use in medicines for the treatment and/or prevention of estrogen receptor-related diseases.
  • Diseases in which the uses described are associated with estrogen receptors include, but are not limited to: cancer (breast cancer, ovarian cancer, colon cancer, prostate cancer, endometrial cancer), osteoporosis, neurodegenerative diseases, cardiovascular disease, lupus erythematosus, endometriosis, and obesity.
  • NMR nuclear magnetic resonance
  • the determination of NMR is to use BRUKER AVANCE III 400 or BRUKER AVANCE III 500 or BRUKER AVANCE III 600 nuclear magnetic resonance spectrometer, and the determination solvent is deuterated dimethyl sulfoxide (DMSO-d 6 ) or deuterated chloroform (CDCl 3 ) or deuterated Substituting methanol (CD 3 OD), the chemical shift is expressed in ⁇ (ppm); mass spectrometry is determined by Finnigan LTQ linear ion trap mass spectrometer or Anggilent UHPLC-QTOF high-resolution mass spectrometer.
  • the silica gel used for separation is 200-300 mesh if not specified, and the ratio of eluent is volume ratio.
  • Step 2 Preparation of 2,3-dibromo-6-((2-(trimethylsilyl)ethoxy)methoxy)benzo[b]thiophene 1,1-dioxide
  • reaction solution was concentrated, then diluted with water (20 mL), extracted with ethyl acetate (20 mL ⁇ 3), the organic phases were combined, washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was distilled off under reduced pressure .
  • Step 5 (2-((3-(4-(2-bromoethoxy)phenoxy)benzo[b]thiophen-6-yl)oxy)methoxy)ethyl)trimethylsilane preparation of
  • Step 7 3-(4-(2-Bromoethoxy)phenoxy)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)benzo[b]thiophene- Preparation of 6-phenol
  • reaction solution was diluted with water (15 mL), extracted with ethyl acetate (15 mL ⁇ 3), the organic phases were combined, washed with saturated sodium chloride solution (15 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was distilled off under reduced pressure.
  • Step 8 3-(4-(2-Bromoethoxy)phenoxy)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)benzo[b]thiophene- Preparation of 6-yltrifluoromethanesulfonate
  • Step 1 2-(3-(4-(2-bromoethoxy)phenoxy)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)benzo[b Preparation of ]thiophen-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
  • Step 2 (3-(4-(2-Bromoethoxy)phenoxy)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)benzo[b]thiophene Preparation of -6-yl)boronic acid
  • Step 3 (2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-(2-(3-(fluoromethyl)azetidine-1 Preparation of -yl)ethoxy)phenoxy)benzothiophen-6-yl)boronic acid
  • 3-Bromobenzothiophene-6-carbonitrile (6585mg, 27.64mmol) was dissolved in ultra-dry dichloromethane (80mL), under the protection of argon, a n-hexane solution of diisobutylaluminum hydride (1M , 30.42mL), then transferred to room temperature for 3h.
  • the reaction solution was cooled to 0°C, diluted with ethyl acetate (100 mL), then slowly added saturated sodium potassium tartrate solution (100 mL), stirred vigorously for 5 minutes, removed the ice bath, and continued to stir until clear.
  • Step 3 Preparation of 3-bromo-6-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)benzo[b]thiophene 1,1-dioxide
  • the reaction solution was filtered to remove excess m-chloroperoxybenzoic acid, the filtrate was quenched with saturated sodium thiosulfate solution, and then extracted with dichloromethane (50mL ⁇ 3), the organic phases were combined, and the organic phase was sequentially washed with saturated sodium carbonate solution ( 50 mL) and saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was distilled off under reduced pressure. The residue was suspended in pure petroleum ether (50 mL), stirred and beaten for 0.5 h, filtered, rinsed with petroleum ether, and dried by infrared to obtain 2528 mg of yellow-white powder with a yield of 82.6%.
  • Step 1 3-(4-(2-Bromoethoxy)phenoxy)-6-(4,4,5,5-tetramethyl-1,3-dioxan-2-yl)benzene Preparation of thiophene 1,1-dioxide
  • Step 2 2-(3-(4-(2-bromoethoxy)phenoxy)benzo[b]thiophen-6-yl)-4,4,5,5-tetramethyl-1,3 - Preparation of dioxolane
  • Step 3 2-(2-Bromo-3-(4-(2-bromoethoxy)phenoxy)benzo[b]thiophen-6-yl)-4,4,5,5-tetramethyl Preparation of -1,3-dioxolane
  • Step 4 2-(3-(4-(2-Bromoethoxy)phenoxy)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)benzo[b Preparation of ]thiophen-6-yl)-4,4,5,5-tetramethyl-1,3-dioxolane
  • Step 5 3-(4-(2-Bromoethoxy)phenoxy)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)benzo[b]thiophene- Preparation of 6-formaldehyde
  • Step 6 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-(2-(3-(fluoromethyl)azetidine-1- Preparation of base) ethoxy) phenoxy) benzothiophene-6-carbaldehyde
  • Step 7 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-(2-(3-(fluoromethyl)azetidine-1- Preparation of base) ethoxy) phenoxy) benzo [b] thiophene-6-carboxylic acid
  • Step 1 Preparation of 3,6-dibromobenzo[b]thiophene 1,1-dioxide:
  • Step 5 Preparation of methyl 3-(4-(benzyloxy)phenoxy)benzo[b]thiophene-6-carboxylate:
  • Step 7 Preparation of methyl 3-(4-(1-tert-butoxycarbonylazetidine-3-oxy)phenoxy)benzo[b]thiophene-6-carboxylate
  • Step 8 Preparation of methyl 2-bromo-3-(4-(1-tert-butoxycarbonylazetidine-3-oxyl)phenoxy)benzo[b]thiophene-6-carboxylate
  • Step 9 3-(4-(1-tert-butoxycarbonylazetidine-3-oxy)phenoxy)-2-((1,1-difluoroethyl)-4-fluorophenyl )
  • Step 9 3-(4-(1-tert-butoxycarbonylazetidine-3-oxy)phenoxy)-2-((1,1-difluoroethyl)-4-fluorophenyl )
  • Step 10 3-(4-(azetidine-3-oxy)phenoxy)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)benzo[ b] Preparation of thiophene-6-carboxylic acid methyl ester hydrochloride
  • Step 11 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3- Preparation of base)oxy)phenoxy)benzothiophene-6-carboxylic acid methyl ester
  • Step 12 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3- Preparation of base)oxy)phenoxy)benzothiophene-6-carboxylic acid
  • Step 1 Preparation of (S)-3-(4-(1-Boc-3-hydroxypyrrolidin-3-oxyl)phenoxy)benzo[b]thiophene-6-carboxylic acid methyl ester
  • Step 2 (S)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)pyrrolidine-3 Preparation of -yl)oxy)phenoxy)benzo[b]thiophene-6-carboxylic acid
  • the preparation method is similar to that of Example 4, except that the raw material 3-(4-(1-tert-butoxycarbonylazetidine-3-oxyl)phenoxy)benzo[ b] Methyl thiophene-6-carboxylate was replaced by (S)-3-(4-(1-tert-butoxycarbonyl-3-hydroxypyrrolidine-3-oxy)phenoxy)benzo[b]thiophene -6-Carboxylic acid methyl ester to give the title compound.
  • Step 1 Preparation of methyl 3-(4-(benzyloxy)phenoxy)-2-bromobenzo[b]thiophene-6-carboxylate
  • Step 3 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-hydroxyphenoxy)benzo[b]thiophene-6-carboxylic acid methyl ester preparation
  • Step 4 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((trifluoromethyl)sulfonyl)oxy)phenoxy)benzo [b] Preparation of methyl thiophene-6-carboxylate
  • Step 5 3-(4-(1-tert-butoxycarbonylazetidine-3-amino)phenoxy)-2-((1,1-difluoroethyl)-4-fluorophenyl) Preparation of benzo[b]thiophene-6-carboxylate methyl ester
  • Step 7 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3- Preparation of (yl) amino) phenoxy) benzothiophene-6-carboxylic acid
  • Step 8 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3- Preparation of base)amino)phenoxy)benzothiophene-6-carboxylic acid
  • Step 2 Preparation of methyl 3-amino-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole-2-carboxylate
  • Step 3 Preparation of methyl 3-bromo-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole-2-carboxylate
  • a mixed solvent 100 mL
  • petroleum ether/ethyl acetate 5/1
  • Step 4 Preparation of 3-bromo-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole-2-carboxylic acid
  • Step 6 Preparation of 3-bromo-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole 1,1-dioxide
  • Step 1 3-(4-(2-Bromoethoxy)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole Preparation of 1,1-dioxide
  • Step 2 3-(4-(2-Bromoethoxy)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole preparation of
  • Step 4 3-(4-(2-Bromoethoxy)phenoxy)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro- Preparation of 2H-pyran-2-yl)-6H-thieno[2,3-e]indazole
  • Step 5 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-(2-(3-(fluoromethyl)azetidine-1- Preparation of base)ethoxy)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole
  • Step 6 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-(2-(3-(fluoromethyl)azetidine-1- Preparation of (yl)ethoxy)phenoxy)-6H-thieno[2,3-e]indazole
  • Example 8 2-(1-(2-(4-((2-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6H-thieno[2,3-e ]indazol-3-yl)oxy) Preparation of phenoxy)ethyl)azetidin-3-yl)acetonitrile
  • Step 1 2-(1-(2-(4-(2-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran-2 Preparation of -yl)-6H-thieno[2,3-e]indazol-3-yl)oxy)phenoxy)ethyl)azetidin-3-yl)acetonitrile
  • Step 2 2-(1-(2-(4-((2-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6H-thieno[2,3-e] Preparation of indazol-3-yl)oxy)phenoxy)ethyl)azetidin-3-yl)acetonitrile
  • the preparation method is similar to the preparation method of Example 7 steps 1-6, except that the raw material 4-(2-bromoethoxy)phenol in step 1 is replaced by 4-(2-bromoethoxy)-2, 6-Difluorophenol to give the title compound.
  • Step 1 Preparation of 3-(4-bromophenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole
  • the preparation method was similar to the preparation method in Step 1-2 of Example 7, except that the raw material 4-(2-bromoethoxy)phenol in Step 1 was replaced with 4-bromophenol to obtain the title compound.
  • Step 3 (E)-3-(4-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran-2-yl )-6H-thieno[2,3-e]indazol-3-yl)oxy)phenyl)methyl acrylate
  • Step 3 (E)-3-(4-((2-(2-( 1,1-Difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-E]indazol-3-yl )oxyl)phenyl)prop-2-en-1-alcohol preparation
  • Step 4 (E)-3-(4-((2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran-2- Preparation of allyl)-6H-thieno[2,3-E]indazol-3-yl)oxy)phenyl)methanesulfonate
  • Step 5 (E)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-(3-(3-(fluoromethyl)azetidine Preparation of alk-1-yl)prop-1-en-1-yl)phenoxy)-6H-thieno[2,3-e]indazole
  • Example 12 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-((2-(3-(fluoromethyl)azetidine- 1-yl)ethoxy) Preparation of methyl)phenoxy)-6H-thieno[2,3-e]indazole
  • Step 1 2-((4-((2-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran-2-yl)- Preparation of 6H-thieno[2,3-e]indazol-3-yl)oxy)benzyl)oxy)ethan-1-ol
  • the preparation method is similar to the preparation method of Example 3 steps 1-4, except that the raw material 4-(2-hydroxyethyl)phenol in step 1 is replaced by 4-((2-hydroxyethoxy)methyl) Phenol, to give the title compound.
  • Step 2 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((2-(3-(fluoromethyl)azetidine-1 Preparation of -yl)ethoxy)methyl )phenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to the preparation method of step 4-5 of Example 11, except that (E)-3-(4-((2-(2-(1,1-difluoroethyl)- 4-fluorophenyl)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-E]indazol-3-yl)oxy)phenyl)propan-2 -en-1-ol was replaced by 2-((4-((2-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran- 2-yl)-6H-thieno[2,3-e]indazol-3-yl)oxy)benzyl)oxy)ethan-1-ol to give the title compound.
  • Example 13 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-((2-(3-(fluoromethyl)azetidine- 1-yl) ethyl) Preparation of sulfonyl)phenoxy)-6H-thieno[2,3-e]indazole
  • Step 1 Preparation of 3-(4-iodophenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole
  • the preparation method was similar to the preparation method in step 1-2 of Example 7, except that 4-(2-bromoethoxy)phenol was replaced by 4-iodophenol to obtain the title compound.
  • Step 2 3-(4-((2-((tert-Butyldimethylsilyl)oxy)ethyl)sulfonyl)phenoxy)-6-(tetrahydro-2H-pyran-2- The preparation of -6H-thieno[2,3-e]indazole
  • Step 3 2-((4-((2-bromo-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazol-3-yl)oxy Base) phenyl) sulfonyl) ethane-1-alcohol preparation
  • Step 4 2-((4-((2-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran-2-yl)- Preparation of 6H-thieno[2,3-e]indazol-3-yl)oxy)phenyl)sulfonyl)ethan-1-ol
  • the preparation method is similar to the preparation method of step 9 of Example 4, except that 3-(4-(1-tert-butoxycarbonylazetidine-3-oxyl)phenoxy)-2-((1 , 1-difluoroethyl)-4-fluorophenyl)benzo[b]thiophene-6-carboxylic acid methyl ester was replaced by 2-((4-((2-bromo-6-(tetrahydro-2H- pyran-2-yl)-6H-thieno[2,3-e]indazol-3-yl)oxy)phenyl)sulfonyl)ethan-1-ol to give the title compound.
  • Step 5 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((2-(3-(fluoromethyl)azetidine-1 -yl)ethyl)sulfonyl)phenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to the preparation method of step 4-5 of Example 11, except that (E)-3-(4-((2-(2-(1,1-difluoroethyl)- 4-fluorophenyl)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-E]indazol-3-yl)oxy)phenyl)propan-2 -en-1-ol was replaced by 2-((4-((2-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran- 2-yl)-6H-thieno[2,3-e]indazol-3-yl)oxy)phenyl)sulfonyl)ethan-1-ol to give the title compound.
  • Example 14 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3 -yl)oxy)benzene Preparation of oxy)-6H-thieno[2,3-e]indazole
  • Step 1 Preparation of 3-(4-(benzyloxy)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to the preparation method of Example 4, step 6, except that 3-(4-(benzyloxy)phenoxy)benzo[b]thiophene-6-carboxylic acid methyl ester is replaced by 3-(4 -(Benzyloxy)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole to give the title compound.
  • Step 3 2-Bromo-3-(4-(1-tert-butoxycarbonylazetidine-3-oxyl)phenoxy)-6-(tetrahydro-2H-pyran-2-yl) Preparation of -6H-thieno[2,3-e]indazole
  • the preparation method is similar to the preparation method of Example 4 Step 7 to Step 8, except that 3-(4-hydroxyphenoxy)benzo[b]thiophene-6-carboxylic acid methyl ester is replaced by 4-((6 -(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazol-3-yl)oxy)phenol to give the title compound.
  • Step 4 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-(1-tert-butoxycarbonylazetidine-3-oxyl)benzene Preparation of oxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole
  • Step 6 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3- Preparation of base)oxy)phenoxy)-6H-thieno[2,3-e]indazole
  • Step 1 (S)-3-(4-(1-Boc-3-hydroxypyrrolidin-3-oxyl)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H -The preparation of thieno[2,3-e]indazole
  • the preparation method is similar to the preparation method of Example 5 step 1, except that 3-(4-hydroxyphenoxy)benzo[b]thiophene-6-carboxylic acid methyl ester is replaced by 4-((6-(tetra Hydrogen-2H-pyran-2-yl)-6H-thieno[2,3-e]indazol-3-yl)oxy)phenol to give the title compound.
  • Step 2 (S)-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)pyrrolidine-3 Preparation of -yl)oxy)phenoxy)-6H-thieno[2,3-e]indazole
  • Example 16 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3 -yl)oxy)-2- Preparation of methoxyphenoxy)-6H-thieno[2,3-e]indazole
  • Example 17 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(3-fluoro-4-((1-(3-fluoropropyl)azacycle butane-3-yl)oxy Preparation of yl)phenoxy)-6H-thieno[2,3-e]indazole
  • Example 18 2-(4-fluoro-2-(1-fluorocyclopropyl)phenyl)-3-(4-((1-(3-fluoropropyl)azetidin-3-yl )oxy)phenoxy
  • the preparation method is similar to that of Example 14, except that 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-4,4,5,5- Tetramethyl-1,3,2-dioxaborolane in place of 2-(4-fluoro-2-(1-fluorocyclopropyl)phenyl)-4,4,5,5-tetramethyl -1,3,2-dioxaborolane to give the title compound.
  • the preparation method is similar to that of Example 14, except that 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-4,4,5,5- Tetramethyl-1,3,2-dioxaborolane in place of 2-(2-(1,1,1-trifluoropropan-2-yl)phenyl)-4,4,5,5- Tetramethyl-1,3,2-dioxaborolane gave the title compound.
  • Step 1 3-(4-(1-tert-butoxycarbonylazetidine-3-amino)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno [2,3-e]indazole
  • Step 2 3-(4-((1-tert-butoxycarbonylazetidin-3-yl)-N-tert-butoxycarbonylamino)phenoxy)-6-(tetrahydro-2H-pyran Preparation of -2-yl)-6H-thieno[2,3-e]indazole
  • Step 3 2-Bromo-3-(4-((1-tert-butoxycarbonylazetidin-3-yl)-N-tert-butoxycarbonylamino)phenoxy)-6-(tetrahydro- Preparation of 2H-pyran-2-yl)-6H-thieno[2,3-e]indazole
  • Step 4 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-tert-butoxycarbonylazetidin-3-yl)- Preparation of N-tert-butoxycarbonylamino)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole
  • Step 5 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3- Preparation of (yl)amino)phenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to the preparation method of step 5-6 in Example 14, except that 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-( 4-(1-tert-butoxycarbonylazetidine-3-oxyl)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3- e] Indazole is replaced by 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-((1-tert-butoxycarbonylazetidine-3- Base)-N-tert-butoxycarbonylamino)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazole to give the title compound .
  • Example 21 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3 -yl)amino)-6- Preparation of fluorophenoxy)-6H-thieno[2,3-e]indazole
  • Example 22 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3 -yl)amino)-6- Preparation of fluorophenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 20, except that the 1-tert-butoxycarbonyl-3-aminoazetidine in step 1 is replaced by (R)-3-aminopyrrolidine-1-carboxylic acid tert-Butyl ester to give the title compound.
  • Example 25 2-(2-(4-fluoro-2-(2,2,2-trifluoroethyl)phenyl)-3-(4-((1-(3-fluoropropyl)azepine cyclobutan-3-yl) ammonia Preparation of base)-phenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 20, except that 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-4,4,5,5- Tetramethyl-1,3,2-dioxaborolane was replaced by 2-(2-(1,1-difluoroethyl)-4,5-difluorophenyl)-4,4,5 ,5-Tetramethyl-1,3,2-dioxaborolane to give the title compound.
  • Example 27 2-((2-(4-fluoro-2-(2-fluoropropan-2-yl)phenyl)-3-(4-((1-(3-fluoropropyl)azacycle Butane-3-yl) ammonia Preparation of yl)phenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 20, except that 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-4,4,5,5- Tetramethyl-1,3,2-dioxaborolane was replaced by 2-((2-(4-fluoro-2-(2-fluoropropan-2-yl)phenyl)-4,4, The title compound was obtained from 5,5-tetramethyl-1,3,2-dioxaborolane.
  • Example 28 2-((2-(4-fluoro-2-(1,1,1,2-tetrafluoropropan-2-yl)phenyl)-3-(4-((1-(3- Fluoropropyl)azetidine-3- Preparation of base)amino)phenoxy)-6H-thieno[2,3-e]indazole
  • Example 30 2-(2-(4-fluoro-2-(2,2,2-trifluoroethyl)phenyl)-3-(4-((1-(3-fluoropropyl)azepine cyclobutan-3-yl) ammonia Preparation of yl)-6-fluorophenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 20, except that 4-iodophenol is replaced by 2-fluoro-4-iodophenol, and 2-(2-(1,1-difluoroethyl) in step 4 -4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was replaced by 2-(2-(4-fluoro-2-(2, 2,2-trifluoroethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane to obtain the title compound.
  • Example 31 2-(2-(1,1-Difluoroethyl)-4,5-difluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine Alk-3-yl) ammonia Preparation of yl)-6-fluorophenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 20, except that 4-iodophenol is replaced by 2-fluoro-4-iodophenol, and 2-(2-(1,1-difluoroethyl) in step 4 -4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane is replaced by 2-(2-(1,1-difluoroethyl) -4,5-difluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane to give the title compound.
  • Example 32 2-(2-(1,1-Difluoroethyl)-4,5-difluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine Alk-3-yl) ammonia Preparation of yl)-6-methoxyphenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 20, except that 4-iodophenol is replaced by 2-methoxy-4-iodophenol, and 2-(2-(1,1-difluoroethane in step 4 yl)-4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was replaced by 2-(2-(1,1-difluoroethyl (yl)-4,5-difluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane to give the title compound.
  • Example 33 2-(2-(4-fluoro-2-(2,2,2-trifluoroethyl)phenyl)-3-(4-((1-(3-fluoropropyl)azepine cyclobutan-3-yl) ammonia Preparation of yl)-6-methoxyphenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 20, except that 4-iodophenol is replaced by 2-methoxy-4-iodophenol, and 2-(2-(1,1-difluoroethane in step 4 Base)-4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was replaced by 2-(2-(4-fluoro-2-( 2,2,2-trifluoroethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane to obtain the title compound.
  • Example 34 2-(2-(4-fluoro-2-(2-fluoropropan-2-yl)phenyl)-3-(4-((1-(3-fluoropropyl)azetidinine Alk-3-yl) ammonia Preparation of yl)-6-methoxyphenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 20, except that 4-iodophenol is replaced by 2-methoxy-4-iodophenol, and 2-(2-(1,1-difluoroethane in step 4 Base)-4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was replaced by 2-((2-(4-fluoro-2- (2-Fluoropropan-2-yl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane gave the title compound.
  • Example 35 2-(2-(1,1-Difluoroethyl)-4,5-difluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine Alk-3-yl)oxy Preparation of yl)-6-methoxyphenoxy)-6H-thieno[2,3-e]indazole
  • Example 36 2-(2-(4-fluoro-2-(2,2,2-trifluoroethyl)phenyl)-3-(4-((1-(3-fluoropropyl)azepine Cyclobutan-3-yl)oxy Preparation of yl)-6-fluorophenoxy)-6H-thieno[2,3-e]indazole
  • Example 37 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3 -yl)oxy)-6- Preparation of fluorophenoxy)-6H-thieno[2,3-e]indazole
  • Example 38 2-(2-(1,1-Difluoroethyl)-4,5-difluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine Alk-3-yl)oxy Preparation of yl)-6-fluorophenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 14, except that the 4-benzyloxyphenol in step 1 is replaced by 2-fluoro-4-benzyloxyphenol, and the 2-(2-(1 ,1-difluoroethyl)-4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was replaced by 2-(2-(1 ,1-difluoroethyl)-4,5-difluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane to give the title compound.
  • Example 39 2-(2-(4-Fluoro-2-(2-fluoropropan-2-yl)phenyl)-3-(4-((1-(3-fluoropropyl)azetidinine Alk-3-yl)oxy Preparation of yl)-6-fluorophenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 14, except that the 4-benzyloxyphenol in step 1 is replaced by 2-fluoro-4-benzyloxyphenol, and the 2-(2-(1 ,1-difluoroethyl)-4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was replaced by 2-(2-(1 ,1-difluoroethyl)-4,5-difluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane to give the title compound.
  • Example 40 2-(2-(1,1-difluoroethyl)-4,5-difluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine Alk-3-yl)oxy Preparation of yl)-6-fluorophenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 14, except that 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-4,4,5,5- Tetramethyl-1,3,2-dioxaborolane was replaced by 2-(2-(1,1-difluoroethyl)-4,5-difluorophenyl)-4,4,5 ,5-Tetramethyl-1,3,2-dioxaborolane to give the title compound.
  • the preparation method is similar to that of Example 14, except that the 4-benzyloxyphenol in step 1 is replaced by 2-methoxyl-4-benzyloxyphenol, and the 2-(2-benzyloxyphenol in step 4 (1,1-difluoroethyl)-4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was replaced by 2-(2- (4-fluoro-2-(2,2,2-trifluoroethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane The title compound.
  • Example 42 2-(2-(4-Fluoro-2-(2-fluoropropan-2-yl)phenyl)-3-(4-((1-(3-fluoropropyl)azetidinine Alk-3-yl)oxy Preparation of yl)-6-methoxyphenoxy)-6H-thieno[2,3-e]indazole
  • Example 43 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3 Preparation of -yl)oxy) phenoxy)-6H-thieno[2,3-e]indazole L-tartrate
  • the freeze-dried powder was suspended in methyl tert-butyl ether (2 mL), stirred at room temperature for 30 min, filtered, and the filter cake was vacuum-dried at 35° C. to obtain 56 mg of white powder with a yield of 80%.
  • Example 45 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3 Preparation of -yl)oxy)phenoxy )-6H-thieno[2,3-e]indazole maleate
  • Example 46 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3 Preparation of -yl)oxy)phenoxy )-6H-thieno[2,3-e]indazole citrate
  • Example 48 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-((1-(3-fluoropropyl)azetidine-3 Preparation of -yl)oxy) phenoxy)-6H-thieno[2,3-e]indazole phosphate
  • the preparation method is similar to that of Example 7, except that the raw material 3-(fluoromethyl)azetidine hydrochloride in step 5 is replaced by 6-fluoro-2-azaspiro[3.3]heptane alkane hydrochloride to give the title compound.
  • Step 1 3-((4-((6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazol-3-yloxy)phenyl)sulfur Substitute) azetidine-1-carboxylate tert-butyl ester
  • Step 2 3-(2-Bromo-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3-e]indazol-3-yl)oxy)phenylthio Generation) Preparation of azetidine-1-carboxylate tert-butyl ester
  • Step 3 3-((4-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran-2-yl)-6H - Preparation of thieno[2,3-e]indazole-3-oxyl)phenyl)thio)azetidine-1-carboxylic acid tert-butyl ester
  • Step 4 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-(1-(3-fluoropropyl)azetidine-3-sulfur Substitute)phenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to the preparation method of step 5-6 in Example 14, except that 2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-( 4-(1-tert-butoxycarbonylazetidine-3-oxyl)phenoxy)-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3- e] Indazole is replaced by 3-((4-(2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-(tetrahydro-2H-pyran-2-yl )-tert-butyl 6H-thieno[2,3-e]indazol-3-oxy)phenyl)thio)azetidine-1-carboxylate to give the title compound.
  • Example 51 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-3-(4-(2-(3-fluoropropyl)-2-azaspiro[3.3 Preparation of ]heptane-6-yl)oxy)phenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 14, except that the raw material 3-iodoazetidine-1-carboxylate tert-butyl ester in step 3 is replaced by 6-iodo-2-azaspiro[3.3] tert-Butyl heptane-2-carboxylate to give the title compound.
  • the preparation method is similar to that of Example 14, except that the raw material 3-iodoazetidine-1-carboxylic acid tert-butyl ester in step 3 is replaced by 6-iodo-2-azaspiro[3.3] Heptane-2-carboxylic acid tert-butyl ester, substituting the starting material 1-fluoro-3-iodopropane in Step 6 for 1-fluoro-2-iodoethane, gave the title compound.
  • 3,4-difluoroindazole-5-carbonitrile (100mg, 0.558mmol) obtained in the previous step was dissolved in dichloromethane (5mL), then p-toluenesulfonic acid monohydrate (11mg, 0.056mmol) and 3, 4-Dihydro-2H-pyran (0.15mL, 1.675mmol), heated to 40°C for 24h. After the reaction, add water (10mL), extract with dichloromethane (10mL ⁇ 3), combine the organic phases, wash the organic phase with saturated sodium chloride solution (10mL), dry over anhydrous sodium sulfate, filter, and distill off the solvent under reduced pressure .
  • Step 3 Preparation of methyl 3-amino-8-fluoro-6-tetrahydropyran-2-yl-6H-thieno[2,3-e]indazole-2-carboxylate
  • Step 4 Preparation of methyl 3-bromo-8-fluoro-6-tetrahydropyran-2-yl-6H-thieno[2,3-e]indazole-2-carboxylate
  • reaction solution was suction filtered, the filter cake was rinsed with water for 2-3 times, and the solid was dried by infrared.
  • Step 6 Preparation of 3-bromo-8-fluoro-6-tetrahydropyran-2-yl-6H-thieno[2,3-e]indazole-1,1-dioxide
  • Step 7 2-(2-(1,1-Difluoroethyl)-4-fluorophenyl)-8-fluoro-3-(4-(1-(3-fluoropropyl)azetidine Preparation of -3-oxy)phenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 14, except that the starting material 3-bromo-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3- e] Indazole 1,1-dioxide was replaced by 3-bromo-8-fluoro-6-tetrahydropyran-2-yl-6H-thieno[2,3-e]indazole 1 obtained in the previous step ,1-Dioxide to give the title compound.
  • Example 54 8-Chloro-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-(1-(3-fluoropropyl)azetidine Preparation of alk-3-oxy)phenoxy)-6H-thieno[2,3-e]indazole
  • Step 2 Preparation of 3-bromo-8-chloro-6-tetrahydropyran-2-yl-6H-thieno[2,3-e]indazole-1,1-dioxide
  • Step 3 8-Chloro-2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-3-(4-(1-(3-fluoropropyl)azetidine -3-oxy)phenoxy)-6H-thieno[2,3-e]indazole
  • the preparation method is similar to that of Example 14, except that the starting material 3-bromo-6-(tetrahydro-2H-pyran-2-yl)-6H-thieno[2,3- e] Indazole-1,1-dioxide is replaced by 3-bromo-8-chloro-6-tetrahydropyran-2-yl-6H-thieno[2,3-e]indazole obtained in the previous step -1,1-dioxide to give the title compound.
  • CTG CellTiter-Glo
  • ATP is a key indicator of metabolism in living cells
  • the homogeneous assay enables cell lysis and the resulting luminescent signal to be proportional to the amount of ATP present, which in turn is directly proportional to the number of cells in culture.
  • MCF-7 cells were cultured with DMEM+10% FBS. The cells were cultured in a 37°C incubator with 5% CO 2 . Cell subculture, recovery and cryopreservation were carried out according to conventional methods. MCF-7 cells grew to about 80% degree of polymerization, trypsinized the cells, inoculated 4 ⁇ 10 3 cells per well into 96-well plates with a seeding volume of 90 ⁇ L per well, and cultured overnight in a 37°C incubator. The next day, add the drug with a concentration gradient, 10 ⁇ L per well. The highest detection concentration of the compound is 10 ⁇ M, 4-fold dilution, 10 concentrations. The 96-well plate was placed in a 5% CO 2 , 37° C.
  • Table 1 In vitro anti-proliferative activity of representative compounds of the present invention on breast cancer MCF-7 cells
  • Example 16 0.93 Example 19 2.23 Example 20 1.13 Example 21 0.75 Example 23 0.58 Example 24 0.61 Example 25 0.82 Example 26 1.12 Example 27 1.02 Example 28 1.74 Example 31 3.09 Example 32 1.67 Example 33 1.7 Example 34 0.66 Example 35 1.15 Example 36 1.44 Example 37 0.77 Example 38 2.83 Example 39 1.44 Example 40 1.57 Example 41 0.55 Example 42 0.48 Example 49 2.26 Example 52 1.18 Example 53 0.89 the the the the the
  • the compound of the present invention has a strong inhibitory activity on the proliferation of MCF-7 breast cancer cells, and the anti-proliferation activity is obviously better than that of the clinical phase I control drug LSZ102, and far better than the clinical phase III control drug SAR439859.
  • the antitumor activities of the compounds of Examples 42, 41, 23, and 24 were respectively 7-fold and 15-fold higher than those of the controls LSZ-102 and SAR439859.
  • MCF-7 Tam1 cells were established by long-term exposure of MCF-7 cells to 4-hydroxytamoxifen, the active metabolite of tamoxifen, accompanied by long-term estrogen deprivation culture conditions. Resistance to aromatase inhibitors.
  • MCF-7 Tam1 cells were subcultured under the conditions of penicillin (final concentration 100 U/mL), streptomycin (final concentration 100 ⁇ g/mL), 10 ⁇ g/mL human insulin, 1 ⁇ M 4-hydroxytamoxifen and 10 %FBS DMEM medium, when the cells are fused to 90%, discard the old medium, wash the cells twice with 2ml PBS, discard the PBS, add 2mL 0.25% trypsin-0.02% EDTA mixed digestion solution, put it under the microscope Observe, about 30s, when the cells become round, quickly add 2ml of complete medium to stop the digestion, gently pipette, and collect the cells.
  • the highest detection concentration of the compound is 10 ⁇ M, 10-fold dilution, 8 concentrations.
  • a 96-well plate suitable for chemiluminescent detection was used to inoculate 100 ⁇ l of cells (1 ⁇ 10 4 cells/ml) per well. The cells in each group were incubated with the medium containing the corresponding concentration of the compound for 5 days according to the experimental grouping. Take out the cell culture plate and equilibrate at room temperature for 10 min.
  • inhibition rate (OD value of control group-OD value of administration group)/OD value of control group.
  • IC 50 of each compound against MCF-7 Tam1 cells was calculated using GraphPad.
  • Example 14 As shown in Figure 1 and Table 2, compared with LSZ102 and SAR439859, Example 14, Example 20 and Example 50 of the present invention all have stronger proliferation inhibitory effects on MCF-7 Tam1. Especially in Example 14 and Example 50, the anti-tumor activity is more than 3 times that of LSZ102 and more than 5 times that of SAR439859, and has a strong ability to resist the proliferation of MCF-7 tamoxifen-resistant breast cancer cells.
  • the extent of ER degradation induced by the compounds described in the present invention was analyzed by cell-based high-content imaging methods.
  • MCF-7 cell culture medium 88% RPMI 1640, 10% FBS, 1% P/S and 1% GlutaMax.
  • MCF-7 seeding medium 88% RPMI 1640, 10% FBS, 1% P/S and 1% GlutaMax.
  • Day 1 Dilute the cell suspension with cell seeding medium to 8.75 ⁇ 10 4 cells/mL, then drop 40 ⁇ L of the cell suspension into each well of the assay plate and place in a 37°C CO 2 incubator.
  • Day 3 Compounds were diluted 4-fold serially, transferred 500 ⁇ L, 10 concentration points. Dilute the compound with 20 ⁇ L of medium, transfer 10 ⁇ L to the plate with Bravo, and incubate in a 37°C incubator for 24 hours.
  • Day 4 Add 50 ⁇ L of 8% paraformaldehyde to the assay plate and let it stand at room temperature for 40 minutes.
  • Example 15 0.69
  • Example 16 0.40
  • Example 20 1.16
  • Example 21 1.28
  • Example 23 1.07
  • Example 24 0.95
  • Example 27 1.35
  • Example 34 1.27
  • Example 37 1.29
  • Example 41 1.78
  • Example 42 1.41
  • the compound of the present invention has a significant degradation effect on the ER ⁇ protein in MCF-7 cells, and the degradation activity is equivalent to or better than that of the clinical phase I drug LSZ102, and far superior to the clinical phase III control drug SAR439859.
  • the compounds of Examples 8, 9, 15, and 16 have 2-4 times higher activity than SAR439859.
  • HEK293/GAL4/ER ⁇ cell line uses molecular cloning method to express fusion protein of estrogen receptor LBD region (compound binding region) and GAL4DBD region (DNA binding region).
  • the ligand activates ER ⁇
  • the ER ⁇ -GAL4 fusion protein initiates the expression of the downstream luciferase gene, and the plate reader detects the chemiluminescent signal.
  • concentration of ER ⁇ ligand stimulation and the chemiluminescent signal were dose-dependent.
  • HEK293/GAL4/ER ⁇ cell suspension Collect the HEK293/GAL4/ER ⁇ cell suspension, centrifuge at 1000 rpm for 5 minutes, remove the supernatant, and use preheated medium (DMEM (no phenol red), 10% carbon-containing serum, that is, 500mL cell culture medium Contains 450mL DMEM, 50mL carbon-adsorbed serum) to resuspend, count and dilute the cell suspension with culture medium, inoculate 40000 cells/well into 96-well cell culture plate, inoculate 80 ⁇ L cell suspension in each well, 37°C, 5% CO 2 incubator, incubate overnight.
  • preheated medium DMEM (no phenol red)
  • 10% carbon-containing serum that is, 500mL cell culture medium Contains 450mL DMEM, 50mL carbon-adsorbed serum
  • % inhibition rate 100-(RFU compound-RFU blank control)/(RFU negative control-RFU blank control) ⁇ 100%.
  • Negative control agonist-treated cells; blank control: cells without agonist treatment, and then use Prism plotting to calculate the IC 50 value of the compound.
  • Table 4 Representative compounds of the present invention have antagonistic activity on wild-type ER ⁇
  • Example 1 1.55 Example 7 0.72
  • Example 14 0.91
  • Example 16 0.61
  • Example 20 1.05
  • Example 21 1.07
  • Example 23 0.60
  • Example 24 0.61
  • Example 25 1.3
  • Example 27 0.97
  • Example 37 0.97
  • Example 41 0.89
  • Example 42 0.75
  • the compound of the present invention has a strong antagonistic effect on wild-type ER ⁇ , which is significantly better than the clinical phase I drug LSZ102 and the clinical phase III drug SAR439859.
  • the antagonistic activity of the compounds of Examples 16, 23, and 24 to wild-type ER ⁇ was increased by more than 10 times compared with LSZ102, and the antagonistic activity of the compounds of Examples 7, 14, 20, 21, 27, 37, 41, and 42 to wild-type ER ⁇ was comparable to that of LSZ102. It is more than 6 times higher than LSZ-102.
  • mutated ER ⁇ has increased affinity for estradiol and decreased affinity for antagonists.
  • SK-BR-3 cell culture medium 89% 1640 without phenol red, 10% charcoal-treated FBS and 1% GlutaMax
  • Day 1 1. Inoculate 80 ⁇ L of cell suspension and 30,000 cells into each well of the assay plate and incubate at 37°C 5% CO 2 for 24 hours.
  • Day 2 Prepare the transfection reagent and let it stand at room temperature for 15 minutes. Add 10 ⁇ L of transfection reagent to each well of the assay plate and incubate at 37° C. in 5% CO 2 for 24 h.
  • Day 3 Add 10 ⁇ L medium (100 nM ⁇ -estradiol, 10 ⁇ L medium) to the assay plate, and incubate at 37° C. with 5% CO 2 for 24 hours.
  • Day 4 1.
  • Table 5 Antagonistic activity of representative compounds of the present invention against ER ⁇ mutants Y537S and D538G in vitro
  • Example 14 and Example 20 have strong antagonistic effects on ER ⁇ mutants Y537S and D538G, wherein compared to SAR43959, Example 14 and Example 20 have 4- 5-fold increase in activity, while Example 14 and Example 20 have comparable antagonistic activity with SAR439859 against the D538G mutant.
  • MCF-7 (Y537S) cells were plated in 6-well plates at 800,000 cells/well, 1 mL per well. On the second day after plating, 1 mL of compound was added, and the final concentration of administration was 1000 nM, 100 nM, 10 nM, 1 nM and 0.1 nM. After drug treatment for 4 hours, cells were collected, total protein was extracted, and BCA was determined. Fully mix the extracted protein supernatant with 5x protein loading buffer, metal bath: 100°C, 10min, after denaturation, cool to room temperature and store at -80°C.
  • Example 14 can induce the degradation of stronger ER ⁇ (Y537S) mutant protein at a lower concentration (10nM-100nM vs100nM-1000nM), indicating that the compound of the present invention has therapeutic endocrine Potential for Drug Resistance in Estrogen Receptor Mutant (Y537S) Breast Cancer.
  • mice/rats were randomly divided into groups according to body weight, fasted without water for 12-14 hours one day before administration, and fed 4 hours after administration.
  • 0.1mL of blood was collected from each animal through the orbit each time, anticoagulated with EDTAK2, and the collection time points were: 0, 5, 15, 30min, 1, 2, 4, 6, 8, 24h after administration of the test substance.
  • the blood samples were placed on ice after collection, and the plasma was centrifuged within 30 minutes (centrifugation conditions: 5000 rpm, 10 minutes, 4°C). Store at –80°C until analysis.
  • Data acquisition and control system software is Analyst1.5.1 software (Applied Biosystem).
  • the peak integration method of the spectrum sample is automatic integration; the ratio of the peak area of the sample to the peak area of the internal standard is used as the index, and the concentration of the sample is used for regression.
  • Regression method linear regression, the weight coefficient is 1/X 2 .
  • Pharmacokinetic parameters were analyzed with WinNonlin Professional v6.3 (Pharsight, USA) using non-compartmental models.
  • Cmax is the measured maximum blood drug concentration
  • the area under the blood drug concentration-time curve AUC (0 ⁇ t) is calculated by the trapezoidal method
  • Tmax is the peak time of blood drug concentration after administration.
  • the compound of the present invention has superior metabolic properties, exhibits good drug exposure and low clearance rate (far lower than 28mL/min/kg reported in the literature of LSZ102), and has a bioavailability of 66.6%. More than 5 times of that, it is predicted that the compound of the present invention can be administered orally to achieve the anti-tumor goal.
  • Tissues were taken 2 hours after administration: plasma, whole blood, brain, breast, ovary, uterus, liver, kidney, and tissues were taken after euthanasia and bloodletting of animals, cleaned and homogenized with 50% methanol at a ratio of 1:4, and the homogenate Stored at -80°C for analysis.
  • Table 7 Tissue distribution of representative compounds of the present invention (Example 47) in female rats
  • Experimental animals SPF-grade female immature SD rats, 21 days old, provided by Changzhou Cavens Experimental Animal Co., Ltd., raised in an SPF-grade feeding environment, the indoor temperature was controlled at 23 ⁇ 2°C, free to eat and drink and water. The total number of animals is 30. Adaptive feeding for 3 days before the experiment.
  • Experimental grouping Blank group: 2% Tween-80/0.5% hydroxymethylcellulose (10 mg/kg) orally every day for 3 consecutive days. Blank+17 ⁇ -ethinyl estradiol group: 17 ⁇ -ethinyl estradiol (0.1 mg/kg) was orally administered every day for 3 consecutive days. 4-Hydroxytamoxifen group: 4-Hydroxytamoxifen group (60 mg/kg) was taken orally every day for 3 consecutive days.
  • Example 14 group take Example 14 (10 mg/kg) orally every day for 3 consecutive days.
  • Example 16 group take Example 16 (10 mg/kg) orally every day for 3 consecutive days.
  • Example 20 group take Example 20 (10 mg/kg) orally every day for 3 consecutive days.
  • the rats were killed by carbon dioxide method, the uterus was dissected, carefully removed irrelevant tissues, washed with D-Hanks solution 2 to 3 times to remove blood, drained and stored, and weighed.
  • HE experiment 1) Cut the fixed uterine tissue into 4 ⁇ m thick slices, and place the slices in an oven for 1 hour; 2) Dewax the dried paraffin slices with conventional xylene, hydrate with descending gradient ethanol, and wash with distilled water; 3 ) Add hematoxylin to stain for 10min-30min, and then wash away the hematoxylin staining solution with running water; 4) 1% hydrochloric acid ethanol fades, see that the slice turns red, the color is lighter, and put it into running water to make it return to blue color.
  • Example 20 Average weight (g) 53.19 54.3 54.59 50.92 57.89 Average wet weight of uterus (mg) 30.64 75.50 59.10 16.94 21.37 Uterine wet weight: body weight 0.06% 0.14% 0.11% 0.03% 0.04%
  • 17 ⁇ -Ethinyl estradiol and 4-hydroxytamoxifen increased uterine wet weight with a tall columnar phenotype of epithelial cells. This is in sharp contrast to the hypocubic phenotype of the epithelium in the blank control and Example 14, confirming that the compound is a complete antagonist with no risk of developing endometrial cancer.
  • Fetal bovine serum (SH30070.03) (FBS, Hyclone, Logan, UT, USA); penicillin (I9532) (Sigma, St.Louis, MO, USA); streptomycin (85886) (Sigma, St.Louis, MO , USA); recombinant human insulin (91077C) (Sigma, St.Louis, MO, USA); EMEM medium (30-2003) (ATCC, Rockville, MD, USA); trypsin (15090046) (Gibco, Grand Island , NY, USA); HBSS (H6648), DMSO (D8418), PEG400 (8074851000), PEG300 (8074841000), PBS (806552), Solutol HS-15 (42966) (Sigma, St.Louis, MO, USA); Matrigel Matrix (BD Bioscience, USA); Estrogen pellets (0.36mg estradiol, 60-day release) (SE-121) (Innovative Research of America, Florida, USA); PVDF membrane (0.45 ⁇
  • MCF-7 cells were subcultured under culture conditions containing penicillin (final concentration 100 U/mL), streptomycin (final concentration 100 ⁇ g/mL), human recombinant insulin (final concentration 0.01 mg/mL) and 10% FBS.
  • EMEM medium when the cells are confluent to 90%, discard the old medium, wash the cells twice with 2ml PBS, discard the PBS, add 2mL 0.25% trypsin-0.02% EDTA mixed digestion solution, observe under the microscope, about After 30s, when the cells become round, quickly add 2ml of complete medium to stop the digestion, and gently pipette to collect the cells. Centrifuge at 800rpm at 4°C for 5min, discard the supernatant, resuspend the cells with complete medium, culture in separate bottles, and change the medium every other day.
  • mice Three days before tumor implantation, estrogen pellets (0.36 mg estradiol, 60 day release) were implanted subcutaneously between the shoulder blades of mice using a sterile trocar. MCF-7 cells in the logarithmic growth phase were taken, digested with trypsin, and resuspended to a cell suspension of 10 7 cells/mL with 50% HBSS and 50% Matrigel mixture. Each mouse was subcutaneously injected with 200 ⁇ L of MCF-7 cell suspension in the right axillary mammary fat pad area, and the tumor volume (width 2 ⁇ length ⁇ ⁇ /6) and body weight were measured every 3 days; when the average tumor volume reached about 200 mm 3 , The mice were randomly divided into groups and administered intragastrically.
  • TGI (%) [(1-(average tumor volume at the end of administration of a certain treatment group-average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of administration of the solvent control group-the beginning of administration of the solvent control group The average tumor volume at the time of drug)] ⁇ 100%.
  • Table 9 Tumor volume of representative compound of the present invention subcutaneous xenograft tumor model
  • the compounds of the present invention can treat or prevent various diseases related to estrogen by antagonizing/degrading estrogen receptors, such as cancer (breast cancer, ovarian cancer, colon cancer, prostate cancer, endometrial cancer), Osteoporosis, neurodegenerative diseases, cardiovascular diseases, lupus erythematosus, endometriosis and obesity, etc.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Epidemiology (AREA)
  • Rheumatology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Cardiology (AREA)
  • Child & Adolescent Psychology (AREA)
  • Dermatology (AREA)
  • Endocrinology (AREA)
  • Reproductive Health (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Obesity (AREA)
  • Hematology (AREA)
  • Diabetes (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

本发明公开了一种苯并噻吩类衍生物、及其制备方法和用途，该苯并噻吩类衍生物结构如通式I所示，各取代基的定义如说明书和权利要求书所述。本发明的苯并噻吩类衍生物，作为选择性雌激素受体拮抗剂/降解剂，用于治疗雌激素受体阳性疾病。

Description

苯并噻吩类衍生物、及其制备方法和用途 技术领域
本发明属于医药领域，具体涉及一类苯并噻吩类衍生物，其外消旋体、对映异构体、非对映异构体及其混合物、其药学上可接受的盐、前体药、含有其的药物组合物，及其制备方法和其在医药上的应用。特别地，本发明涉及通式(I)所示的苯并噻吩类衍生物、其外消旋体、对映异构体、非对映异构体及其混合物、其药学上可接受的盐、前体药、含有其的药物组合物，作为选择性雌激素受体拮抗剂/降解剂的疾病治疗用途，所述的疾病特别是雌激素受体阳性(ER +)疾病。
背景技术
根据世界卫生组织国际癌症研究机构(IARC)发布的2020年全球最新癌症负担数据，2020年全球新发乳腺癌达到226万例，首次超过肺癌(221万例)成为全球第一大癌症。乳腺癌是一种雌激素依赖性肿瘤，雌激素通过与雌激素受体作用促进乳腺癌细胞的增殖。雌激素受体是甾体激素受体，属于核受体超家族，介导了大部分已知的雌激素效应。ER存在两种亚型，其中ERα分布于乳腺、子宫、骨骼、主动脉、肝脏等组织，ERβ则在睾丸、卵巢颗粒细胞、骨髓和大脑中高度表达。研究表明，ERα在约74％乳腺癌患者中高表达，且是乳腺癌细胞增殖的主要驱动因素。除此之外，雌激素作为一种重要的信号分子，参与男性和女性多种组织类型的发育和稳态。雌激素与雌激素受体结合，通过细胞质信号通路或激活细胞核转录，调节基因表达、代谢、细胞生长和增殖等基本功能。雌激素水平的异常表达(过高或过低)已被证明在许多疾病的发病发生和发展中发挥关键作用，包括卵巢癌，结肠癌，前列腺癌，子宫内膜癌，骨质疏松症，神经退行性疾病，心血管疾病，红斑狼疮，子宫内膜异位症及肥胖症。
在ER +的乳腺癌患者中，内分泌治疗是主要的治疗手段。传统的内分泌治疗药物包括芳香化酶抑制剂(AIs)如来曲唑和阿那曲唑(抑制/降低内源性雌激素的生物合成)，以及选择性雌激素受体调节剂(SERMs)如他莫昔芬、雷洛昔芬(与雌激素竞争性结合ER)。尽管内分泌疗法取得了巨大的进步，但是30-50％的患者在内分泌治疗5年内会出现耐药进而发生疾病进展、转移，5年生存率仅20％左右，中位总生存时间也只有2～3年。这些耐药的乳腺癌中，癌细胞的增殖仍然依赖于ERα信号通路的活性。此外，在乳腺癌的发生发展过程中，大约有15％-30％的乳腺癌患者会发生脑转移，并通常伴有内分泌耐药的发生。而常规激素治疗通常无法到达脑组织，不足以产生足够的治疗效果。可以说乳腺癌脑转移是目前乳腺癌治疗中最为棘手的问题，目前还没有发现有效的治疗药物。因此，通过新机制靶向ER来阻断癌细胞增殖信号通路，尤其是开发具有透血脑屏障能力的新型抗雌激素药物具有重要临床价值。
选择性雌激素受体降解剂(SERDs)不仅具有更强的雌激素受体信号通路阻断能力，还能够诱导雌激素受体降解，从而克服因受体突变或过表达导致的耐药性。其中氟维司群是雌二醇7α-烷基化的类似物，与雌激素竞争性结合ER，并将疏水性部分附加到受体的表面， 模拟部分未折叠蛋白状态，利用胞质蛋白未折叠响应(unfolded protein response，UPR)系统来降解靶标ER。氟维司群是目前FDA批准的唯一SERD，每月肌肉注射500/250mg剂量，用于内分泌治疗后复发或疾病进展的ER +局部晚期或转移性绝经后乳腺癌患者，或既往未接受过内分泌治疗的绝经后HR +(孕激素受体阳性)/HER2 -(人表皮生长因子受体-2阴性)局部进展期/转移性乳腺癌患者的治疗。除此之外还有多个候选SERDs药物分子处于临床研究不同阶段，公开结构的选择性雌激素受体降解剂/拮抗剂专利申请包括LSZ-102(US9322746)，G1T48(WO2018148576)、GDC-0927(WO2016097073)、SAR439859(CN108884079)、AZD9833(WO2019002442)、GDC9545(CN107108611)。
Figure PCTCN2022134097-appb-000001
虽然SERDs在乳腺癌等与ER相关的疾病治疗方面显示出令人欣喜的临床疗效，但氟维司群因为口服吸收差，生物利用度低，仅能肌肉注射给药，使得病人的顺应性差，并且给药后需要3-6个月时间来实现平稳的血浆浓度，难以达到抗肿瘤最佳疗效所需的体内药物浓度，并且缺乏透血脑屏障的能力。到目前为止还没有口服有效的小分子SERDs获批上市。
本领域尚需进一步研发获得口服有效、安全性好且顺应性更好，能够透过血脑屏障的ER拮抗剂/降解剂，以满足临床治疗需求。
发明内容
本发明的目的在于提供一类结构新颖、口服有效的强效ER降解剂/拮抗剂(双功能化合物)，用于与ER信号通路相关疾病，特别是乳腺癌、卵巢癌、结肠癌、前列腺癌、子宫内膜癌的治疗。
本发明的第一方面，提供一种通式(I)所示的化合物，或其前药、对映异构体、非对映异构体、外消旋体，或其药学上可接受的盐，
Figure PCTCN2022134097-appb-000002
式中，R 1选自：-COOH、-B(OH) 2、C1-C6烷基、C1-C6卤代烷基；R 2选自：H、卤素、氰基、C1-C6烷基、C1-C6卤代烷基；或者R 1、R 2与相连的苯环形成
Figure PCTCN2022134097-appb-000003
p为0、1、2、3或4；q为0、1、2或3；各R 3、各R 4独立地选自：氢、卤素、氰基、C1-C4烷基、C1-C4烷氧基、C1-C4卤代烷基、C1-C4卤代烷氧基、-SO 2NH 2、-C(O)NH 2；
R 5、R 6各自独立地选自：氢、卤素、C1-C6烷基、C1-C6烷氧基、C1-C6卤代烷基、C1-C6卤代烷氧基，或R 5、R 6与相连的碳形成C3-C6碳环或3-6元杂环；
R 7选自：卤素、C1-C6烷基、C1-C6烷氧基、C1-C6卤代烷基、C1-C6卤代烷氧基；
Z 1为-O-、-NH-、-S-、-CH 2O-、-CO-、-SO 2-、-SO-、-CH＝CH-、-C≡C-、C1-C4亚烷基；
o为0、1、2、3或4；
环Y为
Figure PCTCN2022134097-appb-000004
Z 2、Z 3各自独立地为-CH-或-N-；环A或环B相同或不同，各自独立地为C3-C6环烷基或3-6元杂环烷基；n为0、1、2、3或4；
m为0、1、2、3或4；R 8为卤素、氰基、羟基、羧基或氨基；
R 9选自卤素、氰基、氨基、C1-C6烷基、C1-C6卤代烷基。
在另一优选例中，R 1选自：-COOH、-B(OH) 2、C1-C4烷基、C1-C4卤代烷基；R 2选自：H、卤素、氰基、C1-C4烷基、C1-C4卤代烷基；
或者R 1、R 2与相连的苯环形成
Figure PCTCN2022134097-appb-000005
R 9选自卤素、氰基、氨基、C1-C2烷基、C1-C2卤代烷基。
在另一优选例中，R 5、R 6各自独立地选自：氢、卤素、C1-C4烷基、C1-C4烷氧基、C1-C4卤代烷基、C1-C4卤代烷氧基，或R 5、R 6与相连的碳形成C3-C5碳环或3-5元杂环；
R 7选自：卤素、C1-C4烷基、C1-C4烷氧基、C1-C4卤代烷基、C1-C4卤代烷氧基。
在另一优选例中，Z 1为-O-、-NH-、-S-、-CH 2O-、-CO-、-SO 2-、-SO-、-CH＝CH-、-C≡C-、C1-C2亚烷基。
在另一优选例中，R 1为-COOH、-B(OH) 2；R 2为H；
或者R 1、R 2与相连的苯环形成
Figure PCTCN2022134097-appb-000006
R 9选自氢、氟、氯、氰基、氨基、甲基、三氟甲基；
p为0、1、2或3；各R 3独立地选自：氢、氟、氯、溴、氰基、C1-C2烷基、C1-C2烷氧基、C1-C2卤代烷基、C1-C2卤代烷氧基；
q为0、1或2；各R 4独立地选自：氢、氟、氯、溴；
R 5、R 6各自独立地选自：氢、氟、氯、溴、C1-C3烷基、C1-C2卤代烷基，或R 5、R 6与相连的碳形成C3-C5碳环；
R 7选自：氢、氟、氯、溴、C1-C4烷基、C1-C2卤代烷基；
Z 1为-O-、-NH-、-S-、-CH 2O-、-CO-、-SO 2-、-SO-、-CH＝CH-、-C≡C-、-CH 2-、-CH 2CH 2-；
o为0、1、2或3；
环Y为
Figure PCTCN2022134097-appb-000007
Z 2、Z 3各自独立地为-CH-或-N-；环A或环B相同或不同，彼此独立地代表C4环烷基或4元杂环烷基；n为1、2或3；
m为0、1、2或3；R 8为氟、氯、溴或氰基。
在另一优选例中，R 1为-COOH、-B(OH) 2；R 2为H；或者R 1、R 2与相连的苯环形成
Figure PCTCN2022134097-appb-000008
p为0、1、2或3；各R 3独立地选自：氢、氟、氯、溴、氰基、C1-C2烷基、C1-C2烷氧基、C1-C2卤代烷基、C1-C2卤代烷氧基；
q为0、1或2；各R 4独立地选自：氢、氟、氯、溴；
R 5、R 6各自独立地选自：氢、氟、氯、溴、C1-C3烷基、C1-C2卤代烷基，或R 5、R 6与相连的碳形成C3-C5碳环；R 7选自：氢、氟、氯、溴、C1-C4烷基、C1-C2卤代烷基；
Z 1为-O-、-NH-、-S-、-CH 2O-、-CO-、-SO 2-、-SO-、-CH＝CH-、-C≡C-、-CH 2-、-CH 2CH 2-；
o为0、1、2或3；n为1、2或3；m为0、1、2或3；
Z 2、Z 3各自独立地为-CH-或-N-；
R 8为氟、氯、溴或氰基。
在另一优选例中，环Y为
Figure PCTCN2022134097-appb-000009
在另一优选例中，-CR 5R 6R 7选自：-CF 2CH 3、-CH(CF 3)CH 3、-CH 2CF 3、-CF(CH 3) 2、-CF(CF 3)CH 3、-C(CH 3) 2CF 3、
Figure PCTCN2022134097-appb-000010
在另一优选例中，所述药学上可接受的盐是化合物与有机酸或无机酸形成的盐，所述的无机酸选自盐酸、磷酸、硫酸、氢溴酸、硝酸；所述有机酸选自L-酒石酸、富马酸、L-苹果酸、D-苹果酸、柠檬酸、L-焦谷氨酸、醋酸、对甲苯磺酸盐、苯磺酸、甲磺酸、苯甲酸、乳酸、扁桃酸、马来酸、草酸、丁二酸。
在另一优选例中，所述化合物或其药学上可接受的盐选自实施例1-实施例54制备的任一化合物。
本发明的第二方面，提供第一方面所述的化合物的制备方法，其中，当R 1为-B(OH) 2，R 2为氢时，所述制备方法包括以下步骤：
Figure PCTCN2022134097-appb-000011
(i1)A1与有机硼试剂发生选择性Miyaura偶联反应得到A2，其中，所述的有机硼试剂选自：硼酸、联硼酸频那醇酯、频那醇硼烷、联硼酸新戊二醇酯；
(i2)A2氧化开环得到A3；
(i3)A3与A4发生亲核取代反应得到A5所示的式I化合物；
各式中R 3、R 4、R 5、R 6、R 7、R 8、Z 1、Z 3、m、n、o、p、q如前所述；
X，W分别为离去基团，各自独立地选自：卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基。
或者当R 1为-COOH，R 2为氢时，所述制备方法包括以下步骤：
Figure PCTCN2022134097-appb-000012
Figure PCTCN2022134097-appb-000013
(ii1)B1与B2发生亲核加成消除反应得到B3；
(ii2)B3经过还原溴化得到B4；
(ii3)B4与B5发生Suzuki偶联反应得到B6；
(ii4)B6脱除保护基得到B7；
(ii5)B7与B8发生亲核取代反应得到B9；
(ii6)B9经Pinnick氧化得到B10所示的式I化合物；
各式中R 3、R 4、R 5、R 6、R 7、R 8、Z 1、Z 3、m、n、o、p、q如前所述；
W为离去基团，选自：卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基。
或者当R 1、R 2与相连的苯环形成
Figure PCTCN2022134097-appb-000014
时，所述制备方法包括以下步骤：
Figure PCTCN2022134097-appb-000015
(iii1)中间体int1与C1发生亲核加成消除反应得到C2；
(iii2)C2经过砜基还原得到C3；
(iii3)C3通过催化氢化反应脱除苄基保护基，并随后与C4发生亲核取代反应得到C5；
(iii4)C5经溴化得到C6；
(iii5)C6与C7发生Suzuki偶联得到C8；
(iii6)C8脱除保护基后与C9发生亲核取代反应得到C10所示的式I化合物，
各式中R 3、R 4、R 5、R 6、R 7、R 8、R 9、Z 2、m、n、o、p、q如前所述；
W，X分别为离去基团，各自独立地选自：卤素、三氟甲磺酸酯基、甲磺酸酯基、对 甲苯磺酸酯基；PG为保护基团，选自：叔丁氧羰基、苄氧羰基、芴甲氧羰基、烯丙氧羰基、三氟乙酰基、苄基、对甲氧基苄基、三苯甲基。
或者所述制备方法包括以下步骤：
Figure PCTCN2022134097-appb-000016
(iii1)中间体int1与D1发生亲核加成消除反应得到D2；
(iii2)D2经过还原得到D3；
(iii3)D3与D4发生偶联反应得到D5；
(iii4)D5经溴化得到D6；
(iii5)D6与D7发生Suzuki偶联得到D8；
(iii6)D8脱除保护基后与D9发生亲核取代反应得到D10所示的式I化合物，
在上式中，R 9存在或不存在；
各式中R 3、R 4、R 5、R 6、R 7、R 8、R 9、Z 1、Z 2、m、n、o、p、q、环Y如前所述；
W，X分别为离去基团，各自独立地选自：卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基；PG为保护基团，选自：叔丁氧羰基、苄氧羰基、芴甲氧羰基、烯丙氧羰基、三氟乙酰基、苄基、对甲氧基苄基、三苯甲基。
本发明的第三方面，提供一种药物组合物，包含：第一方面所述的化合物，或其前药、对映异构体、非对映异构体、外消旋体，或其药学上可接受的盐；和药学上可接受的载体。
本发明的第四方面，提供第一方面所述的化合物，或其前药、对映异构体、非对映异构体、外消旋体，或其药学上可接受的盐或第三方面所述的药物组合物的用途，用于制备治疗和/或预防与雌激素受体相关的疾病的药物。在另一优选例中，所述与雌激素受体相关的疾病选自：癌症，骨质疏松症，神经退行性疾病，心血管疾病，红斑狼疮，子宫内膜异位症及肥胖症。在另一优选例中，所述癌症选自：乳腺癌，卵巢癌，结肠癌，前列腺癌，子宫内膜癌。在另一优选例中，所述与雌激素受体相关的疾病选自ER阳性乳腺癌。在另一优选例中，所述与雌激素受体相关的疾病选自ER阳性脑转移性乳腺癌。
应理解，在本发明范围内，本发明的上述各技术特征和在下文(如实施例)中具体描述的 各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。说明书中所揭示的各个特征，可以被任何提供相同、均等或相似目的的替代性特征取代。限于篇幅，在此不再一一赘述。
附图说明
图1示出不同浓度化合物对MCF-7 Tam1细胞增殖抑制作用
图2示出不同浓度化合物对ERα突变体Y537S和D538G的拮抗作用
图3示出不同浓度化合物对MCF-7(Y537S)ERα蛋白降解的作用。
图4为子宫湿重重量实验结果图，其中A示出不同实验组子宫重量与体重的平均比值，*p<0.05,**p<0.01；B为各实验组的子宫横切面组织学图像，子宫内膜表面上皮高度用黑线标示。
图5为化合物对MCF-7小鼠皮下肿瘤模型的生长抑制实验结果图，**p<0.01。
具体实施方式
本申请的发明人经过广泛而深入的研究，开发出一种具有通式I所示结构的化合物，具有ER拮抗和ER降解双重功能，是完全的ER拮抗剂，理论上比具有部分激动活性的SERD能更好的切断ER信号通路，实现对ER +相关疾病更好的治疗效果。此外，本发明化合物具有良好的脑组织暴露量。因此，本发明式(I)化合物适用于治疗和/或预防与雌激素受体相关的疾病或脑转移性疾病的药物中的用途。在此基础上，完成了本发明。
术语
除非有相反陈述，否则下列用在说明书和权利要求书中的属于具有下述含义。
本发明化合物中的“氢”、“碳”、“氧”包括其所有同位素。同位素应理解为包括具有相同原子数但具有不同质量数的哪些原子。举例来说，氢的同位素包括氚和氘，碳的同位素包括 13C和 14C，氧的同位素包括 16O和 18O等；
“卤素”指氟、氯、溴或碘；
“氰基”指-CN
“羧基”指-C(＝O)OH；
“烷基”指仅由C和H两种元素组成的饱和碳氢化合物在任何碳原子失去一个氢原子后形成的基团，包括直链和支链的脂肪烃，非限制性实例包括甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基；
“亚烷基”指具有指定的碳原子数并连接至少两个其他基团的直链或支链饱和脂肪族基团，即二价烃基团。连接到亚烷基的两个基团可以连接到亚烷基上相同的或不同原子。例如，直链亚烷基可以是-(CH2) n-的二价基团，其中n是1、2、3、4、5或6。代表性的亚烷基包括但不限于亚甲基、亚乙基、亚丙基、亚异丙基、亚丁基、亚异丁基、亚仲丁基、亚戊基和亚己基。
“烯基”指由至少两个碳原子和至少一个碳-碳双键组成的如上定义的烷基，例如乙烯基、1-丙烯基、2-丙烯基等；
“环烷基”指饱和或部分不饱和单环或多环环状烃取代基。多环环烷烃包括螺环、稠 环和桥环的环烷基；
“杂环烷基”指包含至少一个(如1、2、3或4个)环杂原子(例如N、O或S)的饱和非芳香性的环状基团，例如氮杂环丁烷基、哌啶基、四氢吡喃基、四氢吡啶基、吡咯啉基、二氢吡啶基、二氢呋喃基、二氢噻吩基、吗啉基；
“螺环基”指两个单环共用一个碳原子的多环基团，其中两个环可以为碳环，也可含有1个或多个杂原子；
“烷氧基”指-O-(烷基)和-O-(环烷基)，其中烷基和环烷基的定义如上所述；
“取代”指基团中的一个或多个氢原子独立地被相应数目的取代基所代替；
“独立地”指当取代基的个数超过一个时，这些取代基可以相同也可以不同；
本发明的“异构体”指具有相同分子式的但在性质上或在其原子的键序列上或在其原子的空间排列上不同的化合物。立体异构体是其原子在空间排列上不同的异构体。彼此不成镜像的立体异构体是非对映体并且互相是非重叠的镜像的立体异构体是对映体。手性化合物可以作为单一的对映体或其混合物存在。包含对映体的相等比例的混合物称作“外消旋混合物”。
本发明的“药学上可接受的盐”指本发明化合物的盐，这类盐用于哺乳动物体内时具有安全性和有效性，且具有应有的生物活性。
具有通式(I)的化合物的体内作用可以部分地由在给予具有通式(I)的化合物之后在人体或动物体内形成的一种或多种代谢物来发挥。如上所述，具有通式(I)的化合物的体内作用也可以经由前体化合物“前体药”代谢来发挥。本发明的“前体药”是指在生物体中的生理条件下，由于与酶、胃酸等反应而转化成式(I)的化合物的化合物，即通过酶的氧化、还原、水解等转化成式(I)的化合物的化合物和/或通过胃酸等的水解反应等转化成式(I)的化合物的化合物。具有羧基的具有通式(I)的化合物的适合的药学上可接受的前体药为例如其体内可裂解的酯。对于羧基的适合的药学上可接受的酯包括烷基酯，如甲基酯、乙基酯和叔丁基酯、烷氧基甲基酯如甲氧基甲基酯；链烷酰氧基甲基酯，如新戊酰氧基酯；3-酞基酯；环烷基羰氧基烷基酯，如环戊基羰氧基甲基酯和1-环己基羰氧基乙基酯；2-氧代-1，3-二氧杂环戊烯基(dioxolenyl)甲基酯，如5-甲基-2-氧代-1，3-二氧杂环戊烯-4-基甲基酯；以及烷氧基羰氧基烷基酯，如甲氧基羰氧基甲基酯和1-甲氧基羰氧基乙基酯。具有羧基的具有通式(I)的化合物的适合的药学上可接受的前药为例如体内可裂解的酰胺，诸如N-烷基酰胺和N，N-二烷基酰胺，如N-甲基酰胺、N-乙基酰胺、N-丙基酰胺、N，N-二甲基酰胺、N-乙基-N-甲基酰胺或N，N-二乙基酰胺。
本发明化合物的制备方法
本发明提供通式(I)所示化合物的通用制备方法。根据以下所描述的方法、以及有机合成化学家通常使用的方法和这些方法的组合或变化来合成通式(I)所述化合物。本发明中的化合物的合成路线不限于以下所总结的方法，个别化合物可能需要调节操作条件以满足各种官能团的要求。本领域技术人员已知的各种保护基团可能是必要的。另外，本文的各个具体实施例也说明了合成本发明化合物的方法。
在本发明化合物通式(I)的一优选例中，R 1为-B(OH) 2；R 2为氢；通式(I)所示的化合物其外消旋体、对映异构体、非对映异构体及其混合物、其药学上可接受的盐、前体药可通过路线1制备。
路线1：
Figure PCTCN2022134097-appb-000017
其中R 3、R 4、R 5、R 6、R 7、R 8、Z 1、Z 3、m、n、o、p、q如前所述；
X，W为离去基团，包括但不限于卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基，优选X为三氟甲磺酸酯基，W为溴；
A2通过A1在钯催化剂、有机硼试剂、碱存在条件下发生选择性Miyaura偶联得到。其中所述的钯催化剂包括但不限于[1,1'-双(二苯基膦)二茂铁]二氯化钯、四(三苯基膦)钯、三(二亚苄基丙酮)二钯、双(二亚芐基丙酮)钯、双三苯基磷二氯化钯、二(三叔丁基膦)钯、双(三环己基膦)钯、醋酸钯；所述的有机硼试剂包括但不限于硼酸、联硼酸频那醇酯、频那醇硼烷、联硼酸新戊二醇酯；所述的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
A3通过A2进行氧化反应得到。所述的氧化剂包括但不限于高碘酸钠；
A5通过A3与A4在碱性条件下发生亲核取代反应得到，所述的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺。
在本发明化合物通式(I)的另一优选例中，通式(I)所述化合物中R 1为-COOH；R 2为氢；通式(I)所示的化合物其外消旋体、对映异构体、非对映异构体及其混合物、其药学上可接受的盐、前体药可通过路线2制备。
路线2：
Figure PCTCN2022134097-appb-000018
Figure PCTCN2022134097-appb-000019
其中R 3、R 4、R 5、R 6、R 7、R 8、Z 1、Z 3、m、n、o、p、q如前所述；
W为离去基团，包括但不限于卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基，优选W为溴；
B3通过B1与B2在碱性条件下发生亲核加成消除反应得到，所述的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
B4通过B3经过砜基还原，芳香溴化得到，其中还原剂包括但不限于二异丁基氢化铝、氢化铝锂、三甲基碘硅烷、二氢双(2-甲氧乙氧基)铝酸钠，硼氢化钠、氰基硼氢化钠、三乙酰氧基硼氢化钠，优选为二异丁基氢化铝；溴化试剂包括但不限于N-溴代丁二酰亚胺、液溴、三溴吡啶嗡盐，优选为N-溴代丁二酰亚胺；
B6通过B4与B5在碱性条件下发生Suziki偶联反应得到，所用钯催化剂包括但不限于[1,1'-双(二苯基膦)二茂铁]二氯化钯、四(三苯基膦)钯、三(二亚苄基丙酮)二钯、双(二亚芐基丙酮)钯、双三苯基磷二氯化钯、二(三叔丁基膦)钯、双(三环己基膦)钯、醋酸钯，所用的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
B7经过B6在酸性条件下的保护基脱除得到，其中所述的酸包括但不限于甲酸、乙酸、甲磺酸、盐酸、硫酸、硝酸、三氟乙酸、三氟甲磺酸、三氯化硼、三溴化硼、三氯化铝；
B9通过B7与B8在碱性条件下发生亲核取代反应得到，所述的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
B10通过B9的pinnick氧化得到。
在本发明化合物通式(I)的另一优选例中，当通式(I)所述化合物中R 1，R 2关环形成吡唑
Figure PCTCN2022134097-appb-000020
时，通式(I)所示的化合物其外消旋体、对映异构体、非对映异构体及其混合物、其药学上可接受的盐、前体药可通过路线3或路线4制备。
路线3：
Figure PCTCN2022134097-appb-000021
在另一优选例中，路线为：
Figure PCTCN2022134097-appb-000022
其中R 9存在或不存在，R 3、R 4、R 5、R 6、R 7、R 8、R 9、Z 2、m、n、o、p、q如前所述；
X，W分别为离去基团，包括但不限于卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基，优选X为碘，W为碘；PG为保护基团，选自：叔丁氧羰基、苄氧羰基、芴甲氧羰基、烯丙氧羰基、三氟乙酰基、苄基、对甲氧基苄基、三苯甲基；
C2从关键中间体int1出发，在碱性条件下与C1发生亲核加成消除反应，所述的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
C3经过C2还原得到，其中还原剂包括但不限于二异丁基氢化铝、氢化铝锂、三甲基碘硅烷、二氢双(2-甲氧乙氧基)铝酸钠，优选为二异丁基氢化铝；
C5通过C3进行催化氢化反应脱除苄基保护剂，其中所用催化剂包括但不限于钯/碳、 氢氧化钯/碳；随后与C4在碱性条件下发生亲核取代反应，所用的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
C6通过C5选择性芳香溴化得到；溴化试剂包括但不限于N-溴代丁二酰亚胺、液溴、三溴吡啶嗡盐，优选为N-溴代丁二酰亚胺；
C8通过C6与C7发生Suzuki偶联得到。其中Suzuki偶联所用钯催化剂包括但不限于[1,1'-双(二苯基膦)二茂铁]二氯化钯、四(三苯基膦)钯、三(二亚苄基丙酮)二钯、双(二亚芐基丙酮)钯、双三苯基磷二氯化钯、二(三叔丁基膦)钯、双(三环己基膦)钯、醋酸钯，所用的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
C10通过C8酸性条件下保护基脱除，进一步与C9在碱性条件下发生亲核取代反应后得到。所述的酸包括但不限于甲酸、乙酸、甲磺酸、盐酸、硫酸、硝酸、三氟乙酸、三氟甲磺酸、三氯化硼、三溴化硼、三氯化铝；所述的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
路线4：
Figure PCTCN2022134097-appb-000023
在另一优选例中，路线为：
Figure PCTCN2022134097-appb-000024
其中R 9存在或不存在；R 3、R 4、R 5、R 6、R 7、R 8、R 9、Z 1、Z 2、m、n、o、p、q如前所述；
X，W分别为离去基团，包括但不限于卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基，优选X为碘，W为碘；；PG为保护基团，选自：叔丁氧羰基、苄氧羰基、芴甲氧羰基、烯丙氧羰基、三氟乙酰基、苄基、对甲氧基苄基、三苯甲基；
D2从关键中间体int1出发，在碱性条件下与D1发生亲核加成消除反应，所述的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
D3经过D2还原得到，其中还原剂包括但不限于二异丁基氢化铝、氢化铝锂、三甲基碘硅烷、二氢双(2-甲氧乙氧基)铝酸钠，优选为二异丁基氢化铝；
D5通过D3与D4发生偶联反应。其中偶联所用钯催化剂包括但不限于[1,1'-双(二苯基膦)二茂铁]二氯化钯、四(三苯基膦)钯、三(二亚苄基丙酮)二钯、双(二亚芐基丙酮)钯、双三苯基磷二氯化钯、二(三叔丁基膦)钯、双(三环己基膦)钯、醋酸钯、氯(2-二环己基膦基-3,6-二甲氧基-2',4',6'-三异丙基-1,1'-联苯)[2-(2-氨基乙基苯基)]钯(II)、甲烷磺酸(2-二环己基膦)-3,6-二甲氧基-2',4',6'-三异丙基-1,1'-联苯)(2'-氨基-1,1'-联苯-2-基)钯(II)，所用的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
D6通过D5选择性芳香溴化得到；溴化试剂包括但不限于N-溴代丁二酰亚胺、液溴、三溴吡啶嗡盐，优选为N-溴代丁二酰亚胺；
D8通过D6与D7发生Suzuki偶联得到。其中Suzuki偶联所用钯催化剂包括但不限于[1,1'-双(二苯基膦)二茂铁]二氯化钯、四(三苯基膦)钯、三(二亚苄基丙酮)二钯、双(二亚芐基丙酮)钯、双三苯基磷二氯化钯、二(三叔丁基膦)钯、双(三环己基膦)钯、醋酸钯，所用的 碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
D10通过D8酸性条件下保护基脱除，进一步与D9在碱性条件下发生亲核取代反应后得到。所述的酸包括但不限于甲酸、乙酸、甲磺酸、盐酸、硫酸、硝酸、三氟乙酸、三氟甲磺酸、三氯化硼、三溴化硼、三氯化铝；所述的碱包括但不限于乙酸钾、氢氧化钾、氢氧化钠、碳酸钠、碳酸钾、磷酸钾、磷酸氢二钾、碳酸铯、叔丁醇钠、叔丁醇钾、甲醇钠、氟化钾、氟化铯、1,8-二氮杂二环[5.4.0]十一碳-7-烯、三乙胺、N,N-二异丙基乙胺；
本发明化合物药学上可接受的盐的制备
Figure PCTCN2022134097-appb-000025
将通式(I)化合物溶于适当溶剂中，1当量酸溶于相同溶剂或水中。随后将酸溶液滴加至通式(I)化合物的溶液中，搅拌30分钟，冻干。将冻干粉末悬浮于甲基叔丁基醚或正己烷中，打浆30分钟，过滤，真空干燥得相应的盐。
其中所用酸为有机酸或无机酸，所述的无机酸包括但不限于盐酸、磷酸、硫酸、氢溴酸、硝酸；有机酸包括但不限于L-酒石酸、富马酸、L-苹果酸、D-苹果酸、柠檬酸、L-焦谷氨酸、醋酸、对甲苯磺酸盐、苯磺酸、甲磺酸、苯甲酸、乳酸、扁桃酸、马来酸、草酸、丁二酸。
其中所述的溶剂包括但不限于乙腈、丙酮、乙酸乙酯、二氯甲烷、四氢呋喃、甲醇、乙醇、异丙醇、1,4-二氧六环、氯仿、乙醚。
含有本发明的化合物的药物组合物和药物剂型
本发明还一种药物组合物，其含有通式(I)的化合物及其外消旋体、对映异构体、非对映异构体及其混合物、其药学上可接受的盐、前体药，与药学上可接受的载体、佐剂或赋形剂相混合。
本发明的药物组合物可根据需要配置成适用于口服、胃肠外(包括皮下、肌肉、皮层和静脉)施予、支气管施予或鼻施予的剂型。其中，优选地，本发明的药物组合物被配置成适用于口服的剂型。
这些组合物还可以包含各种赋形剂，例如，防腐剂、润湿剂、乳化剂和分散剂。
用于口服的固体剂型包括胶囊、药片、药丸、粉末和颗粒。在这些固体剂型中，将活性化合物与至少一种惰性赋形剂(或载体)混合，其中还可以包括：(a)填料或混合剂；(b)粘结剂；(c)保湿剂；(d)崩解剂；(e)溶液阻滞剂；(f)吸收促进剂；(g)润湿剂；(h)吸附剂；(i)润滑剂；
用于口服的液体剂型包括药学上可接受乳液、溶液、分散液、糖浆或酏剂。出了活性化合物意外，液体剂型可以包含本领域中通常使用的惰性稀释剂(例如，水货其他溶剂)、增溶剂和乳化剂。
本发明的化合物的应用
本发明的化合物具有ER拮抗和ER降解双重功能，是完全的ER拮抗剂，理论上比具有部分激动活性的SERD能更好的切断ER信号通路，实现对ER +相关疾病更好的治疗效果。因此，本发明式(I)化合物适用于治疗和/或预防与雌激素受体相关的疾病的药物中的用途。其中所述的用途与雌激素受体相关的疾病包括但不限于：癌症(乳腺癌，卵巢癌，结肠癌，前列腺癌，子宫内膜癌)，骨质疏松症，神经退行性疾病，心血管疾病，红斑狼疮，子宫内膜异位症，及肥胖症等。
下面代表性的实施例是为了更好地说明本发明，而非用于限制本发明的保护范围。以下实施例中使用的材料如无特殊说明均为商购获得。
化合物的结构通过核磁共振(NMR)来确定。NMR的测定是用BRUKER AVANCE III 400或BRUKER AVANCE III 500或BRUKER AVANCE III 600核磁共振波谱仪，测定溶剂为氘代二甲基亚砜(DMSO-d 6)或氘代氯仿(CDCl 3)或氘代甲醇(CD 3OD)，化学位移以δ(ppm)表示；质谱的测定是用Finnigan LTQ线性离子阱质谱仪或Angilent UHPLC-QTOF高分辨质谱仪。分离用硅胶，未说明均为200-300目，洗脱剂的配比均为体积比。
中间体A1的制备
Figure PCTCN2022134097-appb-000026
3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-酚的制备
Figure PCTCN2022134097-appb-000027
步骤1：2,3-二溴-6-羟基苯并[b]噻吩1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000028
将2,3-二溴-6-甲氧基苯并[b]噻吩1,1-二氧化物(5g，14.12mmol)溶于超干二氯甲烷(50mL)，氩气保护，0℃下滴加三溴化硼的二氯甲烷溶液(1M，42.35mL)，随后升至室温，反应24h。将反应液冷至0℃，缓慢滴加甲醇淬灭，加水(100mL)稀释，二氯甲烷(80mL×3)萃取，合并有机相，用饱和氯化钠溶液(100mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。将固体残留物悬浮于石油醚/乙酸乙酯(5/1，50mL)的混合溶液中，搅拌打浆0.5h， 过滤，石油醚(50mL)淋洗固体，真空干燥得淡黄色粉末4.086g，收率85.1％。 1H NMR(400MHz,CDCl 3)δ7.43(dd,J＝8.3,1.1Hz,1H),7.25(d,J＝1.9Hz,2H),7.05(d,J＝7.6Hz,1H).
步骤2：2,3-二溴-6-((2-(三甲基硅基)乙氧基)甲氧基)苯并[b]噻吩1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000029
将上步所得2,3-二溴-6-羟基苯并[b]噻吩1,1-二氧化物(0.4g，1.18mmol)溶于乙腈中(10mL)，加入碳酸钾(0.422g，1.3mmol)，氩气保护，随后滴加2-(三甲基硅烷基)乙氧甲基氯(0.23mL，1.3mmol)，室温反应3h。将反应液浓缩，随后加水(20mL)稀释，乙酸乙酯(20mL×3)萃取，合并有机相，用饱和氯化钠溶液(30mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝50/1)纯化后，浓缩得浅橙色固体437mg，收率78.8％。 1H NMR(400MHz,CDCl 3)δ7.48–7.45(m,2H),7.25(dd,J＝7.9,1.5Hz,1H),5.28(s,2H),3.77–3.72(t,2H,8.2Hz),0.98–0.92(t,2H,8.2Hz),0.00(s,9H).
步骤3：2-溴-3-(4-(2-溴乙氧基)苯氧基)-6-((2-(三甲基硅基)乙氧基)甲氧基)苯并[b]噻吩1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000030
将上步所得2,3-二溴-6-((2-(三甲基硅基)乙氧基)甲氧基)苯并[b]噻吩1,1-二氧化物(1.05g，2.23mmol)溶于四氢呋喃中(20mL)，加入4-(2-溴乙氧基)苯酚(0.58g，2.68mmol)，碳酸铯(2.18g，6.69mmol)，随后室温搅拌过夜。加水(20mL)稀释，乙酸乙酯(20mL×3)萃取，合并有机相，用饱和氯化钠溶液(30mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂，得橙色粗品油状物，不经纯化直接进行下一步。 1H NMR(400MHz,CDCl 3)δ7.50(d,1H),7.41(d,J＝8.5Hz,1H),7.18(dd,J＝8.6,2.2Hz,1H),7.10(d,J＝8.8Hz,2H),6.92(d,J＝8.8Hz,2H),5.28(s,2H),4.30(t,J＝6.2Hz,2H),3.75(d,J＝8.4Hz,2H),3.66(t,J＝6.2Hz,2H),0.96(d,J＝8.1Hz,2H),0.01(t,J＝0.8Hz,9H).
步骤4：3-(4-(2-溴乙氧基)苯氧基)-6-(2-(三甲基硅基)乙氧基)甲氧基)苯并[b]噻吩1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000031
将上一步所得2-溴-3-(4-(2-溴乙氧基)苯氧基)-6-((2-(三甲基硅基)乙氧基)甲氧基)苯并[b]噻吩1,1-二氧化物粗品(2.23mmol)溶于甲醇/二甲基亚砜的混合溶剂中(1/3，20mL)，将反应液冷至0℃，随后加入硼氢化钠(253mg，6.69mmol)，室温搅拌反应5h。0℃下加水(20mL)淬灭反应，乙酸乙酯(20mL×3)萃取，合并有机相，用水(30mL×3)和饱和氯化钠溶液(30mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂，得粗品油状物，不经纯化直接进行下 一步。 1H NMR(400MHz,CDCl 3)δ7.63(d,J＝8.4Hz,1H),7.44(d,J＝2.0Hz,1H),7.23(d,J＝8.7Hz,1H),7.13(d,J＝8.6Hz,2H),6.96(d,J＝8.7Hz,2H),5.41(s,1H),5.29(s,2H),4.31(t,J＝6.2Hz,2H),3.76(d,J＝8.2Hz,2H),3.66(t,J＝6.1Hz,2H),0.96(d,J＝8.2Hz,2H),0.01(s,9H).
步骤5：(2-((3-(4-(2-溴乙氧基)苯氧基)苯并[b]噻吩-6-基)氧基)甲氧基)乙基)三甲基硅烷的制备
Figure PCTCN2022134097-appb-000032
将上步所得3-(4-(2-溴乙氧基)苯氧基)-6-(2-(三甲基硅基)乙氧基)甲氧基)苯并[b]噻吩1,1-二氧化物粗品(2.23mmol)溶于超干四氢呋喃(20mL)和二氯甲烷(10mL)的混合溶剂中，氩气保护，在0℃下滴加二异丁基氢化铝的正己烷溶液(1M，13.4mL)，75℃回流2h。将反应液冷至0℃，加入乙酸乙酯(20mL)稀释，随后缓慢加入饱和酒石酸钾钠溶液(30mL)，剧烈搅拌5分钟，移除冰浴，继续搅拌直至澄清。用乙酸乙酯(20mL×3)萃取，合并有机相，用饱和氯化钠溶液(30mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝50/1-20/1)纯化后，浓缩得黄色油状物388mg，三步收率35.1％。 1H NMR(400MHz,CDCl 3)δ7.65(dd,J＝8.8,0.5Hz,1H),7.48(dd,J＝2.5,0.5Hz,1H),7.10–7.06(m,3H),6.90(d,J＝9.1Hz,2H),6.32(s,1H),5.28(s,2H),4.28(t,J＝6.3Hz,2H),3.81–3.76(m,2H),3.64(t,J＝6.2Hz,2H),1.01–0.96(m,2H),0.01(s,9H).
步骤6：2-溴-3-(4-(2-溴乙氧基)苯氧基)苯并[b]噻吩-6-酚的制备
Figure PCTCN2022134097-appb-000033
将上步所得(2-((3-(4-(2-溴乙氧基)苯氧基)苯并[b]噻吩-6-基)氧基)甲氧基)乙基)三甲基硅烷(380mg，0.77mmol)溶于乙腈中(10mL)，加入N-溴代丁二酰亚(150mg，0.84mmol)，氩气保护，避光条件下，反应5h。饱和硫代硫酸钠溶液(10mL)淬灭反应，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-5/1)纯化后，浓缩得浅紫色油状物300mg，收率87.7％。 1H NMR(400MHz,CDCl 3)δ7.29(d,J＝8.7Hz,1H),7.15(d,J＝2.0Hz,1H),6.89(d,J＝9.1Hz,2H),6.86–6.77(m,3H),4.24(d,J＝6.1Hz,2H),3.62(d,J＝6.1Hz,2H).
步骤7：3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-酚的制备
Figure PCTCN2022134097-appb-000034
将上步所得2-溴-3-(4-(2-溴乙氧基)苯氧基)苯并[b]噻吩-6-酚(800mg，1.8mmol)， 2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷(1030mg，3.6mmol)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(132mg，0.18mmol)，碳酸钾(995mg，7.2mmol)溶于乙二醇二甲醚(10mL)和水(2mL)的混合溶剂中，110℃微波反应25分钟。反应液加水(15mL)稀释，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝30/1-10/1)纯化后，浓缩得橙色油状物1.2g，收率63.7％。 1H NMR(400MHz,CDCl 3)δ7.34–7.26(m,3H),7.23–7.21(m,1H),7.03(td,J＝8.1,2.6Hz,1H),6.84(dd,J＝8.7,2.2Hz,1H),6.79–6.70(m,4H),4.20(t,J＝6.2Hz,2H),3.59(t,J＝6.2Hz,2H),1.88(t,J＝18.5Hz,3H).
步骤8：3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-基三氟甲烷磺酸酯的制备
Figure PCTCN2022134097-appb-000035
将上步所得3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-酚(1.2g，2.29mmol)溶于超干二氯甲烷(15mL)中，氩气保护，0℃下滴加三乙胺(0.48mL，3.44mmol)和三氟甲磺酸酐(0.46mL，2.75mmol)，保持0℃下反应5h。加水(10mL)淬灭反应，用二氯甲烷(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝50/1-20/1)纯化后，浓缩得浅黄油状物1.316g，收率87.9％。 1H NMR(400MHz,CDCl 3)δ7.76(d,J＝2.2Hz,1H),7.52(dd,J＝8.8,0.5Hz,1H),7.37–7.29(m,2H),7.24(dd,J＝8.8,2.2Hz,1H),7.09(td,J＝8.1,2.7Hz,1H),6.81–6.73(m,4H),4.23(t,J＝6.2Hz,2H),3.62(t,J＝6.2Hz,2H),1.92(t,J＝18.5Hz,3H).
实施例1：(2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基) 苯氧基)苯并噻吩-6-基)硼酸的制备
步骤1：2-(3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷的制备
Figure PCTCN2022134097-appb-000036
将3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-基三氟甲烷磺酸酯(580mg，0.887mmol)，联硼酸频那醇酯(248mg，0.976mmol)，[1,1'-双(二苯基膦)二茂铁]二氯化(32mg，0.044mmol)，1,1'-双(二苯基膦)二茂铁(24mg，0.044mmol)，乙酸钾(261mg，2.661mmol)溶于1,4-二氧六环(10mL)中，氩气保护，80℃反应24h。反应液中加水(10mL)，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-5/1)纯 化后，浓缩得无色油状物442mg，收率78.7％。 1H NMR(400MHz,CDCl 3)δ8.29(d,J＝0.9Hz,1H),7.71(dd,J＝8.1,1.0Hz,1H),7.43(dd,J＝8.1,0.8Hz,1H),7.34–7.27(m,2H),7.03(td,J＝8.1,2.8Hz,1H),6.77–6.69(m,4H),4.19(t,J＝6.2Hz,2H),3.58(t,J＝6.2Hz,2H),1.86(t,J＝18.5Hz,3H),1.36(s,12H).
步骤2：(3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-基)硼酸的制备
Figure PCTCN2022134097-appb-000037
将上步所得2-(3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷(400mg，0.632mmol)溶于丙酮(10mL)和水(10mL)的混合溶剂中，加入高碘酸钠(270mg，1.263mmol)，乙酸铵(171mg，2.212mmol)，室温反应16h。反应结束后，向反应液中加入1N盐酸溶液酸化，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-1/1)纯化后，浓缩得黄白色粉末160mg，收率46％。 1H NMR(400MHz,CDCl 3)δ8.73(d,J＝0.9Hz,1H),8.16(dd,J＝8.1,1.0Hz,1H),7.60–7.56(m,1H),7.39–7.32(m,2H),7.08(td,J＝8.2,2.9Hz,1H),6.84–6.71(m,4H),4.21(t,J＝6.2Hz,2H),3.60(t,J＝6.2Hz,2H),1.92(t,J＝18.5Hz,3H).
步骤3：(2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基)苯氧基)苯并噻吩-6-基)硼酸的制备
Figure PCTCN2022134097-appb-000038
将上步所得(3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-基)硼酸(40mg，0.073mmol)溶于乙腈(5mL)中，加入碳酸钾(30mg，0.219mmol)，碘化钾(2mg，0.015mmol)，3-(氟甲基)氮杂环丁烷盐酸盐(18mg，0.145mmol)，80℃回流10h。反应结束后，过滤除盐，加入1N盐酸溶液调pH至8-10，减压浓缩，残留物用制备液相分离(流动相：乙腈/水(1‰三氟乙酸)＝30/70-90/10)。冻干得白色粉末20mg，为目标产物的三氟乙酸盐，收率49％。 1H NMR(600MHz,DMSO-d 6)δ10.31(s,1H),8.35(s,1H),8.25(s,2H),7.73(d,J＝8.2Hz,1H),7.57–7.52(m,1H),7.50(dd,J＝9.9,2.8Hz,1H),7.38(t,J＝8.2Hz,1H),7.26(d,J＝8.1Hz,1H),6.88–6.79(m,4H),4.58(d,J＝46.9Hz,2H),4.23–4.12(m,2H),4.10–4.03(m,2H),4.00–3.87(m,2H),3.56–3.41(m,2H),3.17–3.03(m,1H),1.95(t,J＝19.0Hz,3H).MS(ESI-LR)m/z:560.2(M+1) +.HRMS(ESI):Anal.Calcd for C 28H 27[ 11B]F 4NO 4S,560.1684；found 560.1681.
实施例2：(R)-(2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)吡咯烷-1-基)乙氧基) 苯氧基)苯并[b]噻吩-6-基)硼酸的制备
Figure PCTCN2022134097-appb-000039
以(R)-3-(氟甲基)吡咯烷盐酸盐(56mg，0.4mmol)为原料，按照实施例1中步骤3所述方法制备，得到白色粉末48mg，收率为41.9％。 1H NMR(400MHz,DMSO-d 6)δ8.32(s,1H),7.70(d,J＝8.3Hz,1H),7.56–7.45(m,2H),7.36(td,J＝8.3,2.7Hz,1H),7.28(d,J＝8.1Hz,1H),6.83–6.75(m,4H),4.28(ddd,J＝47.7,6.9,4.1Hz,2H),3.95(t,J＝5.7Hz,2H),2.79–2.69(m,2H),2.61–2.32(m,5H),1.94(t,J＝19.0Hz,3H),1.89–1.77(m,1H),1.43–1.33(m,1H).MS(ESI-LR)m/z:574.3(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 29[ 11B]F 4NO 4S,574.1841；found 574.1833.
中间体B1的制备
Figure PCTCN2022134097-appb-000040
步骤1：3-溴苯并噻吩-6-甲醛的制备
Figure PCTCN2022134097-appb-000041
将3-溴苯并噻吩-6-甲腈(6585mg，27.64mmol)溶于超干二氯甲烷(80mL)，氩气保护，0℃下滴加二异丁基氢化铝的正己烷溶液(1M，30.42mL)，随后转移至室温反应3h。将反应液冷至0℃，加入乙酸乙酯(100mL)稀释，随后缓慢加入饱和酒石酸钾钠溶液(100mL)，剧烈搅拌5分钟，移除冰浴，继续搅拌直至澄清。用乙酸乙酯(50mL×3)萃取，合并有机相，用饱和氯化钠溶液(100mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝50/1-20/1)纯化后，浓缩得黄白色粉末6160mg，收率为92.44％。 1H NMR(400MHz,CDCl 3)δ10.14(s,1H),8.38(d,J＝1.0Hz,1H),7.98(t,J＝1.3Hz,2H),7.72(d,J＝0.7Hz,1H).
步骤2：2-(3-溴苯并噻吩-6-基)-4,4,5,5-四甲基-1,3-二氧杂环乙烷的制备
Figure PCTCN2022134097-appb-000042
将上步所得3-溴苯并噻吩-6-甲醛(2g，8.3mmol)溶于甲苯(20mL)，加入对甲苯磺酸一水合物(79mg，0.415mmol)和频哪醇(2941mg，24.89mmol)，搭建Dean-Stark装置除去反应过程中生成的水，120℃反应4h。将反应液冷至室温，加入饱和碳酸氢钠溶液淬灭，分液，用乙酸乙酯(50mL×3)萃取，合并有机相，用饱和氯化钠溶液(50mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝50/1-20/1)纯化后，浓缩得橙色油状物，收率为98.9％。 1H NMR(400MHz,CDCl 3)δ7.99(s,1H),7.82(d,J＝8.3Hz,1H),7.59(d,J＝8.4Hz,1H),7.44(d,J＝1.4Hz,1H),6.13(s,1H),1.35(s,6H),1.28(s,6H).
步骤3：3-溴-6-(4,4,5,5-四甲基-1,3-二氧戊环-2-基)苯并[b]噻吩1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000043
将上步所得2-(3-溴苯并噻吩-6-基)-4,4,5,5-四甲基-1,3-二氧杂环乙烷(2.8g，8.2mmol)溶于二氯甲烷(30mL)，冷至0℃，15分钟内分批加入间氯过氧苯甲酸(4164mg，20.5mmol)，加毕反应液移至室温，继续反应5h。反应液过滤除去过量的间氯过氧苯甲酸，滤液用饱和硫代硫酸钠溶液淬灭，随后将用二氯甲烷(50mL×3)萃取，合并有机相，有机相依次用饱和碳酸钠溶液(50mL)和饱和氯化钠溶液(50mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。残留物悬浮于纯石油醚中(50mL)，搅拌打浆0.5h，过滤，石油醚淋洗，红外干燥得黄白色粉末2528mg，收率82.6％。 1H NMR(400MHz,CDCl 3)δ7.87(s,1H),7.76(d,J＝7.8Hz,1H),7.53(d,J＝7.8Hz,1H),6.96(d,J＝1.1Hz,1H),5.99(s,1H),1.32(s,6H),1.20(s,6H).
实施例3：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基) 苯氧基)苯并[b]噻吩-6-羧酸的制备
步骤1：3-(4-(2-溴乙氧基)苯氧基)-6-(4,4,5,5-四甲基-1,3-二氧六环-2-基)苯并噻吩1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000044
以3-溴-6-(4,4,5,5-四甲基-1,3-二氧戊环-2-基)苯并[b]噻吩1,1-二氧化物(760mg，2.03mmol)和4-(2-溴乙氧基)苯酚(530mg，2.44mmol)为原料，按照中间体A1制备方法中步骤3所述方法制备，不经纯化，得到棕褐色固体，直接进行下一步。 1H NMR(400MHz,CDCl 3)δ7.91(s,1H),7.75(d,J＝7.9Hz,1H),7.71(d,J＝7.8Hz,1H),7.14(d,J＝8.9Hz,2H),6.97(d,J＝8.9Hz,2H),6.01(s,1H),5.50(s,1H),4.31(t,J＝6.2Hz,2H),3.66(t,J＝6.2Hz,2H),1.33(s,6H),1.23(s,6H).
步骤2：2-(3-(4-(2-溴乙氧基)苯氧基)苯并[b]噻吩-6-基)-4,4,5,5-四甲基-1,3-二氧戊环的制备
Figure PCTCN2022134097-appb-000045
以上步所得3-(4-(2-溴乙氧基)苯氧基)-6-(4,4,5,5-四甲基-1,3-二氧六环-2-基)苯并噻吩1,1-二氧化物(2.03mmol)为原料，按照中间体A1制备方法中步骤5所述方法制备，粗品经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得无色油状物720mg，两步收率为74.3％。 1H NMR(400MHz,CDCl 3)δ7.94(s,1H),7.75(d,J＝8.4Hz,1H),7.49(dd,J＝8.3,1.4Hz,1H),7.10–7.03(m,2H),6.93–6.86(m,2H),6.51(s,1H),6.11(s,1H),4.28(t,J＝6.2Hz,2H), 3.64(t,J＝6.2Hz,2H),1.35(s,6H),1.29(s,6H).
步骤3：2-(2-溴-3-(4-(2-溴乙氧基)苯氧基)苯并[b]噻吩-6-基)-4,4,5,5-四甲基-1,3-二氧戊环的制备
Figure PCTCN2022134097-appb-000046
以上步所得2-(3-(4-(2-溴乙氧基)苯氧基)苯并[b]噻吩-6-基)-4,4,5,5-四甲基-1,3-二氧戊环(720mg，1.51mmol)为原料，按照中间体A1制备方法中步骤6所述方法制备，得浅橙色油状物605mg，收率为72.5％。 1H NMR(400MHz,CDCl 3)δ7.85(s,1H),7.43(d,J＝8.3Hz,1H),7.39(dd,J＝8.3,1.4Hz,1H),6.90–6.85(m,2H),6.85–6.81(m,2H),6.05(s,1H),4.24(t,J＝6.2Hz,2H),3.61(t,J＝6.2Hz,2H),1.34(s,6H),1.27(s,6H).
步骤4：2-(3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-基)-4,4,5,5-四甲基-1,3-二氧戊环的制备
Figure PCTCN2022134097-appb-000047
将上步所得2-(2-溴-3-(4-(2-溴乙氧基)苯氧基)苯并[b]噻吩-6-基)-4,4,5,5-四甲基-1,3-二氧戊环(600mg，1.08mmol)，2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷(617mg，2.16mmol)，四三苯基膦钯(125mg，0.108mmol)，碳酸钾(746mg，5.4mmol)溶于甲苯(10mL)和水(1mL)的混合溶剂中，90℃反应30h。反应液中加水(15mL)，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得橙色油状物540mg，收率为78.7％。 1H NMR(400MHz,CDCl 3)δ7.94(s,1H),7.42(d,J＝1.0Hz,2H),7.35–7.27(m,2H),7.03(td,J＝8.2,2.7Hz,1H),6.78–6.69(m,4H),6.09(s,1H),4.19(t,J＝6.3Hz,2H),3.58(t,J＝6.3Hz,2H),1.86(t,J＝18.5Hz,3H),1.35(s,6H),1.30(s,6H).
步骤5：3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-甲醛的制备
Figure PCTCN2022134097-appb-000048
将上步所得2-(2-溴-3-(4-(2-溴乙氧基)苯氧基)苯并[b]噻吩-6-基)-4,4,5,5-四甲基-1,3-二氧戊环(560mg，0.88mmol)溶于四氢呋喃(10mL)，随后加入4M盐酸(0.66mL)，加热至50℃反应过夜。旋去四氢呋喃，加水(15mL)稀释，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经 柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得浅黄油状物330mg，收率为70％。 1H NMR(400MHz,CDCl 3)δ10.10(s,1H),8.32(s,1H),7.82(d,J＝8.3,1.1Hz,1H),7.57(d,J＝8.3Hz,1H),7.36–7.28(m,2H),7.07(td,J＝8.1,2.8Hz,1H),6.78–6.72(m,4H),4.20(t,J＝6.2,0.8Hz,2H),3.59(t,J＝0.8,6.2Hz,2H),1.90(t,J＝18.5Hz,3H).
步骤6：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基)苯氧基)苯并噻吩-6-甲醛的制备
Figure PCTCN2022134097-appb-000049
以上步所得3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-甲醛(120mg，0.224mmol)和3-(氟甲基)氮杂环丁烷盐酸盐(56mg，0.448mmol)为原料，按照实施例1制备方法中步骤3所述方法制备。旋去乙腈，加水(15mL)稀释，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(二氯甲烷/甲醇＝200/1-50/1)纯化后，浓缩得浅黄油状物120mg，收率为82.1％。 1H NMR(400MHz,CDCl 3)δ10.10(s,1H),8.32(dd,J＝1.4,0.7Hz,1H),7.81(dd,J＝8.3,1.3Hz,1H),7.56(d,J＝8.3Hz,1H),7.35–7.27(m,2H),7.06(td,J＝8.2,2.7Hz,1H),6.77–6.67(m,4H),4.50(dd,J＝47.4,5.6Hz,2H),3.87(t,J＝5.4Hz,2H),3.50(t,J＝7.6Hz,2H),3.15(t,J＝7.0Hz,2H),2.93–2.74(m,3H),1.89(t,J＝18.4Hz,3H).
步骤7：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基)苯氧基)苯并[b]噻吩-6-羧酸的制备
Figure PCTCN2022134097-appb-000050
将上步所得2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基)苯氧基)苯并噻吩-6-甲醛(65mg，0.119mmol)溶于叔丁醇(5mL)，加入四氢呋喃(2mL)助溶，随后加入2-甲基-2-丁烯(80μL，0.952mmol)。将亚氯酸钠(22mg，0.239mmol)和磷酸二氢钠一水合物(82mg，0.595mmol)溶于水(2mL)中，滴加至反应液中，室温反应0.5h。旋去四氢呋喃，加水(15mL)稀释，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品用制备液相分离(流动相：乙腈/水(1‰三氟乙酸)＝30/70-90/10)。冻干得白色粉末28mg，收率42％。 1H NMR(400MHz,DMSO-d 6)δ8.61(d,J＝1.3Hz,1H),7.89(dd,J＝8.4,1.4Hz,1H),7.53(ddd,J＝17.9,9.2,4.2Hz,2H),7.42–7.35(m,2H),6.85–6.74(m,4H),4.50(dd,J＝47.5,6.1Hz,2H),3.85(t,J＝5.4Hz,2H),3.41(t,J＝7.4Hz,2H),3.10(dd,J＝7.7,6.1Hz,2H),2.82–2.68(m,3H),1.95(t,J＝19.0Hz,3H).MS(ESI-LR)m/z:560.3(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 26F 4NO 4S,560.1513；found 560.1518.
实施例4：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基) 苯氧基)苯并[b]噻吩-6-羧酸的制备
步骤1：3,6-二溴苯并[b]噻吩1,1-二氧化物的制备：
Figure PCTCN2022134097-appb-000051
以3,6-二溴苯并噻吩(6.1g，20.89mmol)为原料，按照中间体B1制备方法中步骤3所述方法制备，得黄白色粉末5137mg，收率为75.9％。 1H NMR(400MHz,DMSO-d 6)δ8.34(d,J＝1.8Hz,1H),8.10(s,1H),8.02(dd,J＝8.1,1.7Hz,1H),7.55(d,J＝8.2Hz,1H).
步骤2：3-(4-(苄氧基)苯氧基)-6-溴苯并[b]噻吩1,1-二氧化物的制备：
Figure PCTCN2022134097-appb-000052
将上步所得3,6-二溴苯并[b]噻吩1,1-二氧化物(1.784g，5.51mmol)，4-(苄氧基)苯酚(1211mg，6.05mmol)和碳酸铯(5376mg，16.5mmol)溶于四氢呋喃(50mL)，50℃搅拌过夜。加入等体积石油醚搅拌稀释反应液，硅胶砂芯漏斗过滤除盐，用四氢呋喃/石油醚＝1/1约300mL淋洗固体，减压浓缩滤液，得到油状物粗品。向残留物中加入乙酸乙酯/石油醚＝1/1混合溶剂约100mL，搅拌打浆0.5h，过滤，固体红外干燥，得白色粉末2271mg，收率为93.1％。 1H NMR(400MHz,DMSO-d 6)δ8.29(d,J＝1.7Hz,1H),7.99(dd,J＝8.1,1.8Hz,1H),7.76(d,J＝8.1Hz,1H),7.50–7.45(m,2H),7.44–7.39(m,2H),7.38–7.29(m,3H),7.16–7.11(m,2H),6.23(s,1H),5.13(s,2H).
步骤3：3-(4-(苄氧基)苯氧基)-6-溴苯并噻吩的制备：
Figure PCTCN2022134097-appb-000053
以上步所得3-(4-(苄氧基)苯氧基)-6-溴苯并[b]噻吩1,1-二氧化物(2200mg，2.44mmol)为原料，按照中间体A1制备方法中步骤5所述方法制备，粗品经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得浅黄粉末1771mg，两步收率为87％。 1H NMR(400MHz,CDCl 3)δ7.93(d,J＝1.6Hz,1H),7.67(d,J＝8.6Hz,1H),7.50–7.32(m,6H),7.13–7.06(m,2H),7.01–6.95(m,2H),6.41(s,1H),5.06(s,2H).
步骤4：3-(4-(苄氧基)苯氧基)苯并[b]噻吩-6-羧酸的制备：
Figure PCTCN2022134097-appb-000054
将上步所得3-(4-(苄氧基)苯氧基)-6-溴苯并噻吩(2.6g，6.34mmol)，醋酸钯(43mg，0.19mmol)，4,5-双二苯基膦-9,9-二甲基氧杂蒽(110mg，0.19mmol)，N,N'-二环己基碳二亚胺(262 mg，1.27mmol)加至封管中，加入超干N,N-二甲基甲酰胺(15mL)，氩气保护，在气氛下加入甲酸(1.67ml，44.4mmol)和三乙胺(1.76mL，12.68mmol)，密闭条件下100℃反应20h。反应结束后向反应液中加入水(50mL)，用四氢呋喃/乙酸乙酯＝1/3(60mL×3)萃取，合并有机相，用水(60mL×3)和饱和氯化钠溶液(60mL)依次洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(二氯甲烷/甲醇＝50/1-10/1)纯化后，浓缩得黄色粉末2150mg，收率为86％。 1H NMR(400MHz,DMSO-d 6)δ13.10(s,1H),8.60(s,1H),7.96(dd,J＝8.4,1.5Hz,1H),7.79(d,J＝8.4Hz,1H),7.47–7.30(m,5H),7.17–6.98(m,5H),5.07(s,2H).
步骤5：3-(4-(苄氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯的制备：
Figure PCTCN2022134097-appb-000055
将上步所得3-(4-(苄氧基)苯氧基)苯并[b]噻吩-6-羧酸(2.5g，6.64mmol)和碳酸钾(913mg，6.64mmol)溶于N,N-二甲基甲酰胺(15mL)，随后加入碘甲烷(0.8mL，13.28mL)，80℃反应4h。反应结束后向反应液中加入水(30mL)，用乙酸乙酯(50mL×3)萃取，合并有机相，用水(50mL×3)和饱和氯化钠溶液(50mL)依次洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝30/1-10/1)纯化后，浓缩得白色粉末2g，收率为80.8％。 1H NMR(400MHz,CDCl 3)δ8.52(d,J＝1.5Hz,1H),8.04(dt,J＝8.4,1.4Hz,1H),7.85(d,J＝8.4Hz,1H),7.47–7.37(m,4H),7.37–7.32(m,1H),7.11–7.08(m,2H),7.00–6.96(m,2H),6.63(d,J＝1.2Hz,1H),5.07(s,2H),3.97(s,3H).
步骤6：3-(4-羟基苯氧基)苯并[b]噻吩-6-羧酸甲酯的制备：
Figure PCTCN2022134097-appb-000056
将上步所得3-(4-(苄氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯(1.9g，4.87mmol)溶于四氢呋喃(20mL)，加入氢氧化钯/碳(3.8g，20％wt)，在氢气氛下50℃反应6h，反应液经过硅藻土过滤，四氢呋喃(60mL)淋洗。滤液减压浓缩后得到粗品，不经纯化直接投下一步。 1H NMR(400MHz,CDCl 3)δ8.52(dd,J＝1.5,0.7Hz,1H),8.03(dd,J＝8.4,1.4Hz,1H),7.85(dd,J＝8.4,0.8Hz,1H),7.08–7.03(m,2H),6.88–6.82(m,2H),6.61(s,1H),4.92(s,1H),3.97(s,3H).
步骤7：3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000057
将上步所得3-(4-羟基苯氧基)苯并[b]噻吩-6-羧酸甲酯(4.87mmol)溶于N,N-二甲基甲酰胺(15mL)中，加入1-叔丁氧羰基-3-碘氮杂环丁烷(1654mg，5.844mmol)，碳酸铯(4760mg，14.61mmol)，130℃反应0.5h。反应结束后向反应液中加入乙酸乙酯(50mL)稀释，随后加水(50mL)，用乙酸乙酯(50mL×3)萃取，合并有机相，用水(50mL×3)和饱和氯化钠溶液 (50mL)依次洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝20/1-5/1)纯化后，浓缩得黄白色粉末1940mg，收率为87.4％。 1H NMR(400MHz,CDCl 3)δ8.52(s,1H),8.03(dd,J＝8.4,1.4Hz,1H),7.83(d,J＝8.4Hz,1H),7.12–7.06(m,2H),6.78–6.71(m,2H),6.65(s,1H),4.86(tt,J＝6.3,4.1Hz,1H),4.29(ddd,J＝9.6,6.4,1.0Hz,2H),4.04–3.99(m,2H),3.96(s,3H),1.45(s,9H).
步骤8：2-溴-3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000058
以上步所得3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯(720mg，1.58mmol)为原料，按照中间体A1制备方法中步骤6所述方法制备，得浅黄色油状物260mg，收率为30.8％。 1H NMR(400MHz,CDCl 3)δ8.43(dd,J＝1.4,0.7Hz,1H),7.93(dd,J＝8.4,1.5Hz,1H),7.49(dd,J＝8.4,0.7Hz,1H),6.90–6.84(m,2H),6.69–6.64(m,2H),4.81(tt,J＝6.4,4.1Hz,1H),4.25(ddd,J＝9.7,6.4,1.0Hz,2H),4.00–3.93(m,5H),1.44(s,9H).
步骤9：3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)-2-((1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000059
将上步所得2-溴-3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯(150mg，0.28mmol)，2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷(161mg，0.56mmol)，三(二亚苄基丙酮)二钯(26mg，0.028mmol)，2-二环己基膦-2',4',6'-三异丙基联苯(27mg，0.056mmol)，磷酸钾(178mg，0.84mmol)溶于1,4-二氧六环(5mL)和水(0.5mL)的混合溶剂中，110℃微波反应1.5h。反应结束后，直接将反应液旋干硅胶拌样，经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得橙色油状物100mg，收率为58.2％。 1H NMR(400MHz,CDCl 3)δ8.52(dd,J＝1.5,0.7Hz,1H),7.96(dd,J＝8.4,1.5Hz,1H),7.48(dd,J＝8.5,0.8Hz,1H),7.34–7.27(m,2H),7.05(td,J＝8.1,2.7Hz,1H),6.77–6.73(m,2H),6.58–6.53(m,2H),4.80–4.72(m,1H),4.26–4.20(m,2H),3.97–3.92(m,5H),1.88(t,J＝18.4Hz,3H),1.44(s,9H).
步骤10：3-(4-(氮杂环丁烷-3-氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯盐酸盐的制备
Figure PCTCN2022134097-appb-000060
将上步所得3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)-2-((1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯(110mg，0.161mmol)溶于1,4-二氧六环(5mL)中，加入盐酸的1,4-二氧六环溶液(4M，2mL)，40℃反应过夜。直接将反应液旋干，得粗品不经纯化直接下一步。MS(ESI-LR)m/z:513.2(M+1) +.
步骤11：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯氧基)苯并噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000061
将上步所得3-(4-(氮杂环丁烷-3-氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯盐酸盐(0.16mmol)悬浮于乙腈(5mL)中，加入三乙胺(67μL，0.48mmol)，氩气保护，室温搅拌20分钟后加入1-氟-3-碘丙烷(25μL，0.24mmol)，50℃反应过夜，TLC(二氯甲烷/甲醇＝10/1)监测反应完全并补加1-氟-3-碘丙烷和三乙胺直至反应完全。反应结束后直接向反应液中加入硅胶拌样，经柱层析(二氯甲烷/甲醇＝100/1-100/3)纯化后，浓缩得无色油状物40mg，收率为43.6％。 1H NMR(400MHz,CDCl 3)δ8.53–8.51(m,1H),7.95(dd,J＝8.5,1.5Hz,1H),7.48(d,J＝8.4Hz,1H),7.34–7.27(m,2H),7.05(td,J＝8.1,2.8Hz,1H),6.75–6.70(m,2H),6.60–6.55(m,2H),4.67(p,J＝5.8Hz,1H),4.49(dt,J＝47.2,5.9Hz,2H),3.96(s,3H),3.78(td,J＝6.2,1.9Hz,2H),3.08–3.02(m,2H),2.63(t,J＝7.1Hz,2H),1.88(t,J＝18.5Hz,3H),1.83–1.70(m,2H).
步骤12：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯氧基)苯并噻吩-6-羧酸的制备
Figure PCTCN2022134097-appb-000062
将上步所得2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯氧基)苯并噻吩-6-羧酸甲酯(35mg，0.061mmol)溶于四氢呋喃(5mL)和水(3mL)的混合溶剂中，随后加入氢氧化锂一水合物(8mg，0.183mmol)，室温搅拌过夜。减压浓缩除去四氢呋喃，加水(3mL)稀释，1N盐酸调节pH约为5-6有固体析出。过滤，固体用水淋洗，收集固体后用制备液相分离(流动相：乙腈/水(1‰三氟乙酸)＝30/70-90/10)。流出液旋去大部分乙腈后，饱和碳酸氢钠调节pH至7-8，随后乙酸乙酯萃取，饱和氯化钠洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。残留物冻干得白色粉末16mg，收率为46.9％。 1H NMR (500MHz,DMSO-d 6)δ8.62(s,1H),7.93–7.88(m,1H),7.51(ddd,J＝18.1,9.3,4.2Hz,2H),7.43–7.34(m,2H),6.84–6.76(m,2H),6.72–6.66(m,2H),4.68(p,J＝5.7Hz,1H),4.44(dt,J＝47.4,6.0Hz,2H),3.73(dd,J＝8.2,5.9Hz,2H),2.98(dd,J＝8.3,5.4Hz,2H),2.55(t,J＝7.0Hz,2H),1.95(t,J＝19.0Hz,3H),1.67(dp,J＝25.8,6.5Hz,2H).MS(ESI-LR)m/z:560.4(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 26F 4NO 4S,560.1513；found 560.1517.
实施例5：(S)-2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)吡咯烷-3-基)氧基)苯氧 基)苯并[b]噻吩-6-羧酸的制备
步骤1：(S)-3-(4-(1-Boc-3-羟基吡咯烷-3-氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000063
将实施例4步骤6所得3-(4-羟基苯氧基)苯并[b]噻吩-6-羧酸甲酯(450mg，1.5mmol)，(R)-1-叔丁氧羰基-3-羟基吡咯烷(421mg，2.25mmol)，三丁基膦(1.5mL，6mmol)溶于甲苯(10mL)中，氩气保护，0℃下加入偶氮二甲酰胺(1033mg，6mmol)，随后升温至60℃，搅拌反应过夜。反应结束后向反应液中加入乙酸乙酯(20mL)稀释，随后加水(20mL)，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(20mL)洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝20/1-5/1)纯化后，浓缩得黄白色粉末534mg，收率为75.8％。 1H NMR(400MHz,CDCl 3)δ8.52(d,J＝1.4Hz,1H),8.04(dd,J＝8.4,1.4Hz,1H),7.85(d,J＝8.4Hz,1H),7.12–7.06(m,2H),6.91–6.84(m,2H),6.65(d,J＝4.8Hz,1H),4.86(s,1H),3.97(s,3H),3.70–3.47(m,4H),2.23–2.06(m,2H),1.47(s,9H).
步骤2：(S)-2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)吡咯烷-3-基)氧基)苯氧基)苯并[b]噻吩-6-羧酸的制备
Figure PCTCN2022134097-appb-000064
制备方法类似于实施例4的制备方法，不同的是将步骤8中的原料3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯替换为(S)-3-(4-(1-叔丁氧羰基-3-羟基吡咯烷-3-氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯，制得标题化合物。 1H NMR(400MHz,DMSO-d 6)δ8.60(d,J＝1.4Hz,1H),7.90(dd,J＝8.4,1.4Hz,1H),7.51(ddd,J＝17.0,9.2,4.2Hz,2H),7.41–7.33(m,2H),6.87–6.77(m,2H),6.77–6.71(m,2H),4.76(q,J＝4.9,2.8Hz,1H),4.46(dt,J＝47.3,5.9Hz,2H),2.92(dd,J＝10.8,5.9Hz,1H),2.76(q,J＝7.7Hz,1H),2.67(dd,J＝10.4,2.7Hz,1H),2.61–2.52(m,3H),2.19(dq,J＝14.1,7.3Hz,1H),1.94(t,J＝18.9Hz,3H),1.89–1.68(m,3H).MS(ESI-LR)m/z:574.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 28F 4NO 4S,574.167；found 574.1666.
实施例6：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基)苯 氧基)苯并噻吩-6-羧酸的制备
步骤1：3-(4-(苄氧基)苯氧基)-2-溴苯并[b]噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000065
以实施例4步骤5所得3-(4-(苄氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯(860mg，2.2mmol)为原料，按照中间体A1制备方法中步骤6所述方法制备，粗品经柱层析(石油醚/乙酸乙酯＝50/1-20/1)纯化后，浓缩得浅紫粉末900mg，收率为87.2％。 1H NMR(400MHz,CDCl 3)δ8.45–8.40(m,1H),7.93(dd,J＝8.4,1.5Hz,1H),7.50(dd,J＝8.4,0.7Hz,1H),7.43–7.31(m,5H),6.93–6.86(m,4H),5.01(s,2H),3.95(s,3H).
步骤2：3-(4-(苄氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000066
以上步所得3-(4-(苄氧基)苯氧基)-2-溴苯并[b]噻吩-6-羧酸甲酯(90mg，0.192mmol)为原料，按照实施例4制备方法中步骤9所述方法制备，粗品经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得黄色油状物70mg，收率为66.5％。 1H NMR(400MHz,CDCl 3)δ8.52(t,J＝1.0Hz,1H),7.96(dd,J＝8.4,1.5Hz,1H),7.49(d,J＝8.5Hz,1H),7.42–7.27(m,7H),7.04(td,J＝8.1,2.7Hz,1H),6.82–6.69(m,4H),4.97(s,2H),3.96(s,3H),1.88(t,J＝18.5Hz,3H).
步骤3：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-羟基苯氧基)苯并[b]噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000067
以上步所得3-(4-(苄氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯(1g，1.82mmol)为原料，按照实施例4制备方法中步骤6所述方法制备，滤液减压浓缩后得到粗品，不经纯化直接投下一步。 1H NMR(400MHz,CDCl 3)δ8.52(d,J＝1.0Hz,1H),7.95(dd,J＝8.4,1.5Hz,1H),7.49(d,J＝8.5Hz,1H),7.31(ddd,J＝9.7,8.2,4.1Hz,2H),7.05(td,J＝8.1,2.7Hz,1H),6.73–6.62(m,4H),4.91(s,1H),3.96(s,3H),1.88(t,J＝18.5Hz,3H).
步骤4：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((三氟甲基)磺酰基)氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000068
将上步所得2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-羟基苯氧基)苯并[b]噻吩-6-羧酸甲酯(1.82mmol)溶于超干二氯甲烷(15mL)，随后氩气保护，0℃下加入吡啶(0.22mL，2.73mmol)，再缓慢滴加三氟甲磺酸酐(0.37Ml,2.18mmol)，。反应结束后向反应液中加入水(15mL)淬灭，用二氯甲烷(15mL×3)萃取，合并有机相，依次用1N盐酸(20mL)和饱和氯化钠溶液(20mL)洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝30/1)纯化后，浓缩得无色油状物938mg，收率为87.3％。 1H NMR(400MHz,CDCl 3)δ8.55(d,J＝1.4Hz,1H),8.01(dd,J＝8.5,1.5Hz,1H),7.49(d,J＝8.4Hz,1H),7.33–7.27(m,2H),7.12–7.03(m,3H),6.90–6.84(m,2H),3.97(s,3H),1.89(t,J＝18.4Hz,3H).
步骤5：3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氨基)苯氧基)-2-((1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000069
将上步所得2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((三氟甲基)磺酰基)氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯(295mg，0.5mmol)，1-叔丁氧羰基-3-氨基氮杂环丁烷(172mg，1mmol)，
甲烷磺酸(2-二环己基膦)-3,6-二甲氧基-2',4',6'-三异丙基-1,1'-联苯)(2'-氨基-1,1'-联苯-2-基)(45mg，0.05mmol)，碳酸铯(407mg，1.25mmol)溶于1,4-二氧六环(5mL)，氩气保护，微波110℃反应6h，反应结束后向反应液中加入水(15mL)，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(20mL)洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-3/1)纯化后，浓缩得白色泡沫153mg，收率为49.9％。 1H NMR(400MHz,CDCl 3)δ8.51(dd,J＝1.5,0.7Hz,1H),7.95(dd,J＝8.4,1.5Hz,1H),7.49(d,J＝8.5Hz,1H),7.31(ddd,J＝9.6,6.3,4.0Hz,2H),7.05(td,J＝8.1,2.7Hz,1H),6.72–6.66(m,2H),6.38–6.30(m,2H),4.23(dd,J＝8.9,7.1Hz,2H),4.15–4.04(m,1H),3.96(s,3H),3.80(d,J＝6.8Hz,1H),3.67(dd,J＝9.1,4.7Hz,2H),1.88(t,J＝18.5Hz,3H),1.43(s,9H).
步骤6：3-(4-(氮杂环丁烷-3-氨基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯盐酸盐的制备
Figure PCTCN2022134097-appb-000070
将上步所得3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氨基)苯氧基)-2-((1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯(125mg，0.204mmol)溶于异丙醇(5mL)，随后加入盐酸(2mL，5M in IPA)，50℃搅拌反应过夜。反应完全应结束后，直接将反应液旋干，得粗品不经纯化直接下一步。MS(ESI-LR)m/z:513.1(M+1) +.
步骤7：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基)苯氧基) 苯并噻吩-6-羧酸的制备
Figure PCTCN2022134097-appb-000071
将上步所得3-(4-(氮杂环丁烷-3-氨基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯盐酸盐(0.204mmol)悬浮于乙腈中(10mL)，随后加入碳酸钾(70mg，0.51mmol)，氩气保护，室温搅拌20分钟。之后向反应液中依次加入三乙胺(71μL，0.51mmol)和1-氟-3-碘丙烷(31μL，0.306mmol)，50℃搅拌反应过夜，TLC(二氯甲烷/甲醇＝10/1)监测反应完全并补加1-氟-3-碘丙烷和三乙胺直至反应完全。反应结束后向反应液中加水(15mL)，用乙酸乙酯(15mL×3)萃取，合并有机相，饱和氯化钠洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(二氯甲烷/甲醇＝100/1-100/3)纯化后，浓缩得黄色油状物112mg，收率为95.9％。 1H NMR(400MHz,CDCl 3)δ8.51(d,J＝1.5Hz,1H),7.94(dd,J＝8.5,1.4Hz,1H),7.48(d,J＝8.4Hz,1H),7.35–7.26(m,2H),7.06(td,J＝8.2,2.7Hz,1H),6.73–6.64(m,2H),6.45–6.37(m,2H),4.52(dt,J＝47.0,5.7Hz,2H),4.22(s,1H),4.02(t,J＝8.0Hz,2H),3.95(s,3H),3.47–3.32(m,2H),2.97(t,J＝7.4Hz,2H),2.03–1.82(m,5H).
步骤8：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基)苯氧基)苯并噻吩-6-羧酸的制备
Figure PCTCN2022134097-appb-000072
将上步所得2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基)苯氧基)苯并噻吩-6-羧酸(105mg，0.184mmol)为原料，按照实施例4制备方法中步骤12所述方法制备，得浅黄白色粉末68mg，收率为66.2％。 1H NMR(600MHz,DMSO-d 6)δ8.59(s,1H),7.88(d,J＝8.4Hz,1H),7.54(dd,J＝8.6,5.6Hz,1H),7.49(dd,J＝9.9,2.8Hz,1H),7.42–7.36(m,2H),6.66(d,J＝8.6Hz,2H),6.36(d,J＝8.4Hz,2H),5.82(d,J＝6.8Hz,1H),4.44(dt,J＝47.4,6.0Hz,2H),3.84(q,J＝6.1Hz,1H),3.63(t,J＝6.8Hz,2H),3.48–3.41(m,0H),2.75(t,J＝6.8Hz,2H),1.94(t,J＝19.0Hz,3H),1.65(dp,J＝25.8,6.6Hz,2H).MS(ESI-LR)m/z:559.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 27F 4N 2O 3S,559.1673；found 559.1678.
中间体int1，3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000073
步骤1：4-氟-1-(四氢-2H-吡喃-2-基)-1H-吲唑-5-甲腈的制备
Figure PCTCN2022134097-appb-000074
将5-溴-4-氟-1-(四氢-2H-吡喃-2-基)-1H吲唑(13.35g，44.63mmol)，氰化锌(5.24g，44.63mmol)和四三苯基膦钯(5.157g，4.463mmol)溶于N,N-二甲基甲酰胺(150mL)，氩气保护下100℃反应6h。反应结束后向反应液中加入乙酸乙酯(50mL)稀释，硅藻土过滤，滤饼用乙酸乙酯(150mL淋洗)。随后向滤液中加水(150mL)，用乙酸乙酯(100mL×3)萃取，合并有机相，用水(150mL×3)和饱和氯化钠溶液(150mL)依次洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-5/1)纯化后，浓缩得白色粉末8g，收率为73.1％。 1H NMR(400MHz,CDCl 3)δ8.18(s,1H),7.50–7.48(m,2H),5.74(dd,J＝8.9,2.7Hz,1H),4.02–3.94(m,1H),3.80–3.69(m,1H),2.55–2.41(m,1H),2.20–2.07(m,2H),1.85–1.64(m,3H).
步骤2：3-氨基-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-2-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000075
将上步所得的4-氟-1-(四氢-2H-吡喃-2-基)-1H-吲唑-5-甲腈(8g，32.6mmol)溶于N,N-二甲基甲酰胺(100mL)，加入碳酸钾(18.023g，130.4mmol)和巯基乙酸甲酯(3.2mL，35.86mmol)，升温至65℃，搅拌反应15h。反应结束后，将反应液倾倒至搅拌的冰水中(200mL)，析出大量固体，继续搅拌10分钟。随后将反应液抽滤，滤饼用水淋洗2-3遍，固体红外干燥，得黄白色粉末8.564g，收率为79.3％。 1H NMR(400MHz,CDCl 3)δ8.18(d,J＝0.7Hz,1H),7.59(d,J＝9.2Hz,1H),7.54(d,J＝9.0Hz,1H),5.78(dd,J＝9.2,2.7Hz,1H),4.07–3.99(m,1H),3.90(s,3H),3.81–3.74(m,1H),2.63–2.51(m,1H),2.26–2.04(m,2H),1.85–1.66(m,3H).
步骤3：3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-2-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000076
将上步所得的3-氨基-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-2-羧酸甲酯(10.9g，32.9mmol)，CuBr 2(11.02g，49.35mmol)溶于乙腈(100mL)中，氩气保护下升温至65℃，随后在加热条件下滴加亚硝酸叔丁酯(5.8mL，49.35mmol)，继续加热搅拌反应45分钟。反应结束后将反应液冷却至室温，倒至冰水中(200mL)，搅拌析出大量固体。随后将反应液抽滤，滤饼用水淋洗2-3遍，固体红外干燥。将固体悬浮于石油醚/乙酸乙酯＝5/1的混合溶剂(100mL)中，搅拌打浆0.5h，倾析除去上层清液，固体重复打浆步骤3次后，抽滤，滤饼用石油醚淋洗，红外干燥，得黄绿色粉末8.4g，收率为64.6％。 1H NMR(400MHz,DMSO-d 6)δ8.57(s,1H),7.98(dd,J＝9.2,0.8Hz,1H),7.90(d,J＝9.1Hz,1H),6.00(dd,J＝9.6,2.3Hz,1H),3.91(s,3H),3.91–3.87(m,1H),3.83–3.75(m,1H),2.47–2.37(m,1H),2.09–1.98(m,2H),1.83–1.72(m,1H),1.64–1.56(m,2H).
步骤4：3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-2-羧酸的制备
Figure PCTCN2022134097-appb-000077
将上步所得的3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-2-羧酸甲酯(7.905g，20mmol)溶于四氢呋喃(100mL)和水(60mL)的混合溶剂中，随后加入氢氧化锂一水合物(2517mg，60mmol)，室温搅拌过夜。反应结束后，减压浓缩除去大部分四氢呋喃，残留物中加水(100mL)稀释，在冰浴下用4N盐酸调pH至3-4，有大量固体析出。随后将反应液抽滤，滤饼用水淋洗2-3遍，固体红外干燥，得灰白色粉末7.5g，收率为98.4％。 1H NMR(400MHz,DMSO-d 6)δ13.80(s,1H),8.55(s,1H),7.95(d,J＝9.1Hz,1H),7.87(d,J＝9.1Hz,1H),5.98(dd,J＝9.6,2.3Hz,1H),3.94–3.84(m,1H),3.82–3.74(m,1H),2.46–2.38(m,1H),2.09–1.96(m,2H),1.85–1.69(m,1H),1.65–1.57(m,2H).
步骤5：3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000078
将上步所得的3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-2-羧酸(7624mg，20mmol)，氧化亚铜(1717mg，12mmol)溶于N,N-二甲基甲酰胺(100mL)，140℃加热搅拌反应2h。反应结束后，冷至室温，加入乙酸乙酯(50mL)稀释，硅藻土过滤，滤饼用乙酸乙酯淋洗(100mL)。随后向滤液中加水(100mL)，用乙酸乙酯(50mL×3)萃取，合并有机相，用水(100mL×3)和饱和氯化钠溶液(100mL)依次洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得浅橙色粉末4g，收率为59.3％。 1H NMR(400MHz,CDCl 3)δ8.20(d,J＝0.9Hz,1H),7.80(d,J＝9.0Hz,1H),7.68(dd,J＝8.9,0.8Hz,1H),7.36(s,1H),5.81(dd,J＝9.3,2.7Hz,1H),4.04(ddd,J＝10.2,3.9,2.2Hz,1H),3.83–3.73(m,1H),2.67–2.53(m,1H),2.23–2.08(m,2H),1.86–1.65(m,3H).
步骤6：3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000079
将上步所得的3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(5.3g，15.71mmol)为原料，按照中间体B1制备方法中的步骤3制备，得黄白色粉末5.462g，收率为94.2％。 1H NMR(400MHz,CDCl 3)δ8.34(d,J＝0.9Hz,1H),7.92(dd,J＝8.7,0.9Hz,1H),7.57(d,J＝8.7Hz,1H),6.93(s,1H),5.83(dd,J＝8.4,2.8Hz,1H),3.97–3.90(m,1H),3.80–3.71(m,1H),2.56–2.46(m,1H),2.20–2.09(m,2H),1.84–1.68(m,3H).
实施例7：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基) 苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：3-(4-(2-溴乙氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000080
以3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑1,1-二氧化物(1216mg，3.29mmol)和4-(2-溴乙氧基)苯酚(857mg，3.95mmol)为原料，按照中间体A1制备方法中步骤3所述方法制备，不经纯化，得到土黄色固体，直接进行下一步。 1H NMR(400MHz,CDCl 3)δ8.35(d,J＝0.9Hz,1H),7.92(dd,J＝8.7,0.9Hz,1H),7.75(d,J＝8.7Hz,1H),7.21–7.15(m,2H),7.01–6.96(m,2H),5.84(dd,J＝8.4,2.7Hz,1H),5.46(s,1H),4.32(t,J＝6.2Hz,2H),3.99–3.93(m,1H),3.81–3.72(m,1H),3.67(t,J＝6.2Hz,2H),2.59–2.48(m,1H),2.22–2.09(m,2H),1.87–1.70(m,3H).
步骤2：3-(4-(2-溴乙氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000081
以上步所得得到的3-(4-(2-溴乙氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑1,1-二氧化物(3.29mmol)为原料，按照中间体A1制备方法中步骤5所述方法制备，粗品经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得黄色泡沫状固体1117mg，两步收率为71.7％。 1H NMR(400MHz,CDCl 3)δ8.20(d,J＝0.8Hz,1H),7.74(d,J＝8.9Hz,1H),7.58(dd,J＝9.0,0.9Hz,1H),7.14–7.07(m,2H),6.95–6.87(m,2H),6.39(s,1H),5.79(dd,J＝9.4,2.7Hz,1H),4.28(t,J＝6.2Hz,2H),4.08–4.01(m,1H),3.81–3.73(m,1H),3.64(t,J＝6.2Hz,2H),2.67–2.54(m,1H),2.24–2.06(m,2H),1.85–1.66(m,3H).
步骤3：2-溴-3-(4-(2-溴乙氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000082
以上步所得3-(4-(2-溴乙氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(424mg，0.896mmol)为原料，按照中间体A1制备方法中步骤5所述方法制备，得浅黄色油状物343mg，收率为69.8％。 1H NMR(400MHz,CDCl 3)δ8.16(d,J＝0.9Hz,1H),7.50(dd,J＝9.0,0.9Hz,1H),7.41(d,J＝9.0Hz,1H),6.94–6.87(m,2H),6.86–6.81(m,2H),5.75(dd,J＝9.2,2.7Hz,1H),4.24(t,J＝6.2Hz,2H),4.01(d,J＝11.7Hz,1H),3.80–3.70(m,1H),3.61(t,J＝6.2Hz,2H),2.63–2.51(m,1H),2.25–2.07(m,2H),1.85–1.63(m,3H).
步骤4：3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000083
以上步所得2-溴-3-(4-(2-溴乙氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(330mg，0.598mmol)为原料，按照实施例3中步骤4所述方法制备，得到浅黄色泡沫固体130mg，收率为34.4％。 1H NMR(400MHz,CDCl 3)δ8.21(d,J＝0.8Hz,1H),7.53(dd,J＝9.0,0.9Hz,1H),7.40(d,J＝9.0Hz,1H),7.37–7.31(m,2H),7.04(td,J＝8.3,2.9Hz,1H),6.79–6.69(m,4H),5.77(dd,J＝9.3,2.7Hz,1H),4.20(t,J＝6.2Hz,2H),4.03(d,J＝11.6Hz,1H),3.79–3.71(m,1H),3.59(t,J＝6.2Hz,2H),2.64–2.53(m,1H),2.24–2.07(m,2H),1.87(t,J＝18.5Hz,3H),1.80–1.63(m,3H).
步骤5：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000084
以上步所得的3-(4-(2-溴乙氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(130mg，0.205mmol)和3-(氟甲基)氮杂环丁烷盐酸盐(52mg，0.412mmol)为原料，按照实施例1制备方法中步骤3所述方法制备。粗品经柱层析(二氯甲烷/甲醇＝200/1-50/1)纯化后，浓缩得浅黄油状物120mg，收率为91.5％。 1H NMR(400MHz,CDCl 3)δ8.20(s,1H),7.52(d,J＝9.0Hz,1H),7.40(d,J＝8.9Hz,1H),7.37–7.30(m,2H),7.04(td,J＝8.1,2.8Hz,1H),6.78–6.66(m,4H),5.77(dd,J＝9.3,2.7Hz,1H),4.50(dd,J＝47.4,5.1Hz,2H),4.03(d,J＝11.5Hz,1H),3.93(t,J＝5.3Hz,2H),3.80–3.70(m,1H),3.65(t,J＝7.9Hz,2H),3.28(t,J＝7.4Hz,2H),3.01–2.87(m,3H),2.65–2.51(m,1H),2.21–2.05(m,2H),1.86(t,J＝18.5Hz,3H),1.81–1.62(m,3H).
步骤6：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000085
将上步所得的2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(120mg，0.188mmol)溶于甲醇(5mL)，随后加入盐酸的甲醇溶液(0.4mL，4M)，室温搅拌反应过夜，TLC(二氯甲烷/甲醇＝10/1)监测反应完全。旋去甲醇，加水(10mL)稀释，饱和碳酸氢钠调节pH至8-10，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品用制备液相分离(流动相：乙腈/水(1‰三氟乙酸)＝30/70-90/10)，用碳酸氢钠溶液游离后。冻干得白色粉末50mg，收率47.9％。 1H NMR(600MHz,DMSO-d 6)δ13.51(s,1H),8.39(s,1H),7.58–7.52(m,2H),7.49(dd,J＝9.9,2.8Hz,1H),7.38(td,J＝8.3,2.8Hz,1H),7.25(d,J＝8.9Hz,1H),6.84–6.79(m,2H),6.79–6.75(m,2H),4.49(dd,J＝47.6,6.2Hz,2H),3.81(t,J＝5.6Hz,2H),3.30(t,J＝7.3Hz,2H),2.99(t,J＝6.6Hz,2H),2.75– 2.65(m,3H),1.95(t,J＝19.0Hz,3H).MS(ESI-LR)m/z:556.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 26F 4N 3O 2S,556.1676；found 556.1692.
实施例8：2-(1-(2-(4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基) 苯氧基)乙基)氮杂环丁-3-基)乙腈的制备
步骤1：2-(1-(2-(4-(2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯氧基)乙基)氮杂丁-3-基)乙腈的制备
Figure PCTCN2022134097-appb-000086
制备方法类似于实施例7的步骤5，不同的是将步骤5中的原料3-(氟甲基)氮杂环丁烷盐酸盐替换为3-(氰甲基)氮杂环丁烷盐酸盐，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.20(d,J＝0.9Hz,1H),7.52(dd,J＝9.0,0.9Hz,1H),7.40(d,J＝9.0Hz,1H),7.38–7.28(m,2H),7.04(td,J＝8.1,2.7Hz,1H),6.78–6.70(m,2H),6.73–6.65(m,2H),5.77(dd,J＝9.3,2.7Hz,1H),4.03(d,J＝11.7Hz,1H),3.86(t,J＝5.3Hz,2H),3.75(t,J＝9.7Hz,1H),3.53(t,J＝7.6Hz,2H),3.08(dd,J＝7.6,6.0Hz,2H),2.84–2.74(m,3H),2.62(d,J＝7.4Hz,2H),2.59–2.52(m,1H),2.22–2.07(m,2H),1.86(t,J＝18.5Hz,3H),1.78–1.64(m,3H).
步骤2：2-(1-(2-(4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯氧基)乙基)氮杂环丁-3-基)乙腈的制备
Figure PCTCN2022134097-appb-000087
将上步得到的2-(1-(2-(4-(2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯氧基)乙基)氮杂丁-3-基)乙腈(105mg，0.162mmol)溶于二氯甲烷(5mL)，随后滴加三氟乙酸(0.24mL，3.23mmol)，室温搅拌反应过夜。旋去三氟乙酸，加水(10mL)稀释，饱和碳酸氢钠调节pH至8-10，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品用制备液相分离(流动相：乙腈/水(1‰三氟乙酸)＝30/70-90/10)，用碳酸氢钠溶液游离后。冻干得白色粉末27mg，收率27.6％。 1H NMR(400MHz,DMSO-d 6)δ13.53(s,1H),8.39(s,1H),7.58–7.52(m,2H),7.49(dd,J＝9.3,2.8Hz,1H),7.37(td,J＝8.4,2.7Hz,1H),7.25(d,J＝8.9Hz,1H),6.85–6.79(m,2H),6.79–6.73(m,2H),3.81(t,J＝5.5Hz,2H),3.36(t,J＝10.6Hz,2H),2.90(t,J＝6.7Hz,2H),2.73(d,J＝6.8Hz,2H),2.69–2.58(m,3H),1.95(t,J＝19.0Hz,3H).MS(ESI-LR)m/z:563.3(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 26F 3N 4O 2S,563.1723；found 563.1722.
实施例9：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(2-氟-4-(2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧 基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000088
制备方法类似于实施例7的制备方法，不同的是将步骤1中的原料4-(2-溴乙氧基)苯酚替换为4-(2-溴乙氧基)-2-氟苯酚，制得标题化合物。 1H NMR(400MHz,DMSO-d 6)δ13.54(s,1H),8.39(s,1H),7.59(dd,J＝8.9,1.0Hz,1H),7.53–7.44(m,2H),7.39–7.32(m,2H),6.87(dd,J＝12.9,2.9Hz,1H),6.76(t,J＝9.4Hz,1H),6.50(ddd,J＝9.1,3.0,1.4Hz,1H),4.49(dd,J＝47.5,6.1Hz,2H),3.84(t,J＝5.4Hz,2H),3.35–3.28(m,2H),3.03(t,J＝6.8Hz,2H),2.80–2.65(m,3H),1.93(t,J＝19.0Hz,3H). 19F NMR(471MHz,DMSO-d 6)δ-73.46,-81.72(q,J＝19.0Hz),-110.71–-111.15(m),-131.11(t,J＝11.4Hz).MS(ESI-LR)m/z:574.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 25F 5N 3O 2S,574.1582；found 574.1584.
实施例10：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(2，6-二氟-4-(2-(3-(氟甲基)氮杂环丁烷-1- 基)乙氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000089
制备方法类似于实施例7步骤1-6的制备方法，不同的是将步骤1中的原料4-(2-溴乙氧基)苯酚替换为4-(2-溴乙氧基)-2，6-二氟苯酚，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.55(s,1H),8.37(s,1H),7.66(s,2H),7.46–7.11(m,3H),6.57(d,J＝11.5Hz,2H),4.50(d,J＝47.6Hz,2H),3.83(s,2H),3.28(s,2H),2.96(s,2H),2.77–2.55(m,3H),1.84(t,J＝18.9Hz,3H).MS(ESI-LR)m/z:592.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 24F 6N 3O 2S,592.1491；found 592.1488.
实施例11：(E)-2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(3-(3-(氟甲基)氮杂环丁烷-1-基)丙-1- 烯-1-基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：3-(4-溴苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000090
制备方法类似于实施例7步骤1-2的制备方法，不同的是将步骤1中的原料4-(2-溴乙氧基)苯酚替换为4-溴苯酚，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.21(d,J＝0.8Hz,1H),7.64–7.55(m,2H),7.47–7.40(m,2H),7.03–6.96(m,2H),6.64(s,1H),5.79(dd,J＝9.3,2.7Hz,1H),4.04(d,J＝11.4Hz,1H),3.81–3.72(m,1H),2.67–2.53(m,1H),2.23–2.07(m,2H),1.86–1.64(m,3H).
步骤2：(E)-3-(4-((6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-E]吲唑-3-基)氧基)苯基)丙烯酸甲酯的制备
Figure PCTCN2022134097-appb-000091
将上步得到的3-(4-溴苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(250mg，0.582mmol)，丙烯酸甲酯(0.42mL，4.658mmol)，双三苯基膦二氯化钯(82mg，0.116mmol)，三乙胺(0.48ml，3.492mmol)溶于N’,N’-二甲基甲酰胺(5mL)，氩气保护，微波120℃反应1h。反应结束后，加水(10mL)，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝20/1-5/1)纯化后，浓缩得黄色粉末125mg，收率为49.4％。 1H NMR(400MHz,CDCl 3)δ8.22(d,J＝0.7Hz,1H),7.67(d,J＝16.0Hz,1H),7.60–7.55(m,2H),7.51–7.48(m,2H),7.10–7.07(m,2H),6.76(s,1H),6.35(d,J＝16.0Hz,1H),5.78(dd,J＝9.3,2.8Hz,1H),4.06–4.01(m,1H),3.80(s,3H),3.79–3.73(m,1H),2.63–2.55(m,1H),2.21–2.09(m,2H),1.83–1.73(m,2H),1.69–1.65(m,1H).
步骤3：(E)-3-(4-(2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯基)丙烯酸甲酯的制备
Figure PCTCN2022134097-appb-000092
以上步得到的(E)-3-(4-((6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-E]吲唑-3-基)氧基)苯基)丙烯酸甲酯为原料，制备方法类似于实施例3的步骤3至4，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.22(d,J＝0.8Hz,1H),7.59(d,J＝16.0Hz,1H),7.54(dd,J＝9.0,0.9Hz,1H),7.41–7.31(m,5H),7.05(td,J＝8.1,2.7Hz,1H),6.88–6.82(m,2H),6.28(d,J＝16.0Hz,1H),5.78(dd,J＝9.3,2.6Hz,1H),4.06–3.99(m,1H),3.78(s,3H),3.77–3.70(m,1H),2.65–2.52(m,1H),2.23–2.07(m,2H),1.88(t,J＝18.5Hz,3H),1.80–1.64(m,1H),0.91–0.80(m,2H).步骤3：(E)-3-(4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-E]吲唑-3-基)氧基)苯基)丙-2-烯-1-醇的制备
Figure PCTCN2022134097-appb-000093
将上步得到的(E)-3-(4-(2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯基)丙烯酸甲酯(180mg，0.304mmol)溶于超干二氯甲烷(10mL)，冷至-78℃，随后滴加二异丁基氢化铝(1.06mL，1M in hexane)，滴加完毕，保持该温度反应30分钟，随后缓慢升温至0℃继续反应1h。0℃下，加乙酸乙酯(10mL)稀释，随后加入 饱和酒石酸钾钠(20mL)搅拌至澄清，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝5/1-1/1)纯化后，浓缩得黄白色膏状物79mg，收率为46％。 1H NMR(400MHz,DMSO-d 6)δ8.22(d,J＝1.5Hz,1H),7.52(d,J＝9.0Hz,1H),7.41–7.31(m,3H),7.20(dd,J＝7.1,5.2Hz,2H),7.03(td,J＝8.2,2.6Hz,1H),6.82–6.76(m,2H),6.51(d,J＝15.9Hz,1H),6.25–6.16(m,1H),5.77(dd,J＝9.3,2.5Hz,1H),4.27(d,J＝5.8Hz,2H),4.02(d,J＝11.3Hz,1H),3.79–3.71(m,1H),2.58(q,J＝12.2,11.2Hz,1H),2.22–2.07(m,2H),1.95–1.62(m,6H).
步骤4：(E)-3-(4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-E]吲唑-3-基)氧基)苯基)甲磺酸烯丙基酯的制备
Figure PCTCN2022134097-appb-000094
将上步得到的(E)-3-(4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-E]吲唑-3-基)氧基)苯基)丙-2-烯-1-醇(100mg，0.177mmol)溶于超干二氯甲烷(5mL)，氩气保护，0℃下加入三乙胺(49μL，0.354mmol)和甲磺酰氯(21μL，0.266mmol)，随后室温反应3h。0℃下，加水(10mL)淬灭反应，用二氯甲烷(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品不经纯化，直接下一步。
步骤5：(E)-2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(3-(3-(氟甲基)氮杂环丁烷-1-基)丙-1-烯-1-基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000095
以上步得到的(E)-3-(4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-E]吲唑-3-基)氧基)苯基)甲磺酸烯丙基酯为原料，按照实施例7制备方法中步骤5-6所述方法制备。得白色粉末20mg，收率95.4％。 1H NMR(500MHz,DMSO-d 6)δ13.55(s,1H),8.41(s,1H),7.58–7.46(m,3H),7.35(dt,J＝8.5,4.8Hz,1H),7.31–7.27(m,2H),7.23(d,J＝8.9Hz,1H),6.87–6.83(m,2H),6.41(d,J＝15.9Hz,1H),6.00(dt,J＝15.9,6.2Hz,1H),4.49(dd,J＝47.6,6.1Hz,2H),3.26(t,J＝7.3Hz,2H),3.11(d,J＝5.9Hz,2H),2.95(t,J＝6.6Hz,2H),2.71(dt,J＝18.8,6.2Hz,1H),1.96(t,J＝19.0Hz,3H).MS(ESI-LR)m/z:552.0(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 26F 4N 3O 2S,552.1727；found 552.1717.
实施例12：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基) 甲基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：2-((4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e] 吲唑-3-基)氧基)苄基)氧基)乙烷-1-醇的制备
Figure PCTCN2022134097-appb-000096
制备方法类似于实施例3步骤1-4的制备方法，不同的是将步骤1中的原料4-(2-羟乙基)苯酚替换为4-((2-羟基乙氧基)甲基)苯酚，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.21(s,1H),7.52(d,J＝9.0Hz,1H),7.40–7.30(m,3H),7.16(d,J＝8.3Hz,2H),7.03(td,J＝8.2,2.7Hz,1H),6.82(d,J＝8.1Hz,2H),5.77(dd,J＝9.4,2.7Hz,1H),4.45(s,2H),4.02(d,J＝11.4Hz,1H),3.78–3.71(m,3H),3.56(t,J＝4.5Hz,2H),2.66–2.49(m,1H),2.21–2.07(m,2H),1.88(t,J＝18.5Hz,3H),1.81–1.69(m,3H).
步骤2： 2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((2-(3-(氟甲基)氮杂环丁烷-1-基)乙氧基)甲 基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000097
制备方法类似于实施例11步骤4-5的制备方法，不同的是将步骤4中的(E)-3-(4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-E]吲唑-3-基)氧基)苯基)丙-2-烯-1-醇替换为2-((4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苄基)氧基)乙烷-1-醇，制得标题化合物。 1H NMR(400MHz,Methanol-d 4)δ8.26(s,1H),7.46(d,J＝8.9Hz,1H),7.41–7.32(m,3H),7.15(d,J＝8.5Hz,2H),7.08(dt,J＝8.3,4.2Hz,1H),6.83–6.76(m,2H),4.50–4.32(m,4H),3.43–3.37(m,4H),3.08(ddd,J＝9.9,6.9,2.8Hz,2H),2.77(dt,J＝20.4,6.7Hz,1H),2.62(t,J＝5.4Hz,2H),1.88(t,J＝18.6Hz,3H).MS(ESI-LR)m/z:570.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 28F 4N 3O 2S,570.1833；found 570.1842.
实施例13：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((2-(3-(氟甲基)氮杂环丁烷-1-基)乙基) 磺酰基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：3-(4-碘苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000098
制备方法类似于实施例7步骤1-2的制备方法，不同的是将4-(2-溴乙氧基)苯酚替换为4-碘苯酚，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.21(s,1H),7.65–7.55(m,4H),6.91–6.85(m,2H),6.66(s,1H),5.78(dd,J＝9.3,2.7Hz,1H),4.04(d,J＝11.4Hz,1H),3.81–3.72(m,1H),2.59(q,J＝11.8,10.8Hz,1H),2.23–2.06(m,2H),1.83–1.66(m,3H).
步骤2：3-(4-((2-((叔丁基二甲基硅基)氧基)乙基)磺酰基)苯氧基)-6-(四氢-2H-吡喃-2- 基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000099
将上步得到的3-(4-碘苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(380mg，0.8mmol)，焦亚硫酸钠(304mg，1.6mmol)，(2-溴乙氧基)(叔丁基)二甲基硅烷(576mg，2.4mmol)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(60mg，0.08mmol)，1,1'-双(二苯基膦)二茂铁(88mg，0.16mmol)，锡粉(284mg，2.4mmol)，磷酸氢二钾一水合物(364mg，1.6mmol)和四丁基溴化铵(388mg，1.2mmo)溶于二甲基亚砜(10mL)，氩气保护，100℃加热反应10h，，TLC(石油醚/乙酸乙酯＝5/1)监测反应。反应结束后硅藻土过滤，滤饼用乙酸乙酯(20mL)淋洗，加水，乙酸乙酯(20mL×3)萃取，合并有机相，依次用水(30mL×3)和饱和氯化钠溶液(30mL)洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得浅黄色油状物360mg，收率为78.6％。 1H NMR(400MHz,CDCl 3)δ8.23(d,J＝0.8Hz,1H),7.87–7.82(m,2H),7.57(dd,J＝8.9,0.8Hz,1H),7.48(d,J＝8.9Hz,1H),7.17–7.13(m,2H),6.89(s,1H),5.79(dd,J＝9.3,2.7Hz,1H),4.06–3.97(m,3H),3.76(ddd,J＝11.4,10.0,3.0Hz,1H),3.35(t,J＝6.3Hz,2H),2.64–2.53(m,1H),2.23–2.07(m,2H),1.84–1.67(m,3H),0.80(s,9H),-0.02(s,6H).
步骤3：2-((4-((2-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯基)磺酰基)乙烷-1-醇的制备
Figure PCTCN2022134097-appb-000100
将上步得到的3-(4-((2-((叔丁基二甲基硅基)氧基)乙基)磺酰基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(410mg，0.715mmol)溶于乙腈(5mL)中，随后加入N-溴代丁二酰亚胺(140mg，0.787mmol)，氩气保护。避光条件下，室温搅拌过夜，，TLC(石油醚/乙酸乙酯＝5/1)监测反应。饱和硫代硫酸钠溶液(10mL)淬灭反应，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(15mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝5/1-1/2)纯化后，浓缩得白色泡沫状固体196mg，收率51％。 1H NMR(400MHz,CDCl 3)δ8.19(d,J＝0.8Hz,1H),7.90–7.85(m,2H),7.55(dd,J＝9.0,0.9Hz,1H),7.36(d,J＝8.9Hz,1H),7.13–7.08(m,2H),5.77(dd,J＝9.2,2.7Hz,1H),4.05–3.97(m,3H),3.80–3.69(m,1H),3.39–3.31(m,2H),2.62–2.50(m,1H),2.23–2.05(m,2H),1.84–1.67(m,3H).
步骤4：2-((4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯基)磺酰基)乙烷-1-醇的制备
Figure PCTCN2022134097-appb-000101
制备方法类似于实施例4步骤9的制备方法，不同的是将3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)-2-((1,1-二氟乙基)-4-氟苯基)苯并[b]噻吩-6-羧酸甲酯替换为2-((4-((2-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯基)磺酰基)乙烷-1-醇，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.22(d,J＝0.9Hz,1H),7.78–7.75(m,2H),7.58(dd,J＝8.9,0.9Hz,1H),7.36–7.30(m,3H),7.07(td,J＝8.1,2.7Hz,1H),7.03–6.99(m,2H),5.79(dd,J＝9.2,2.8Hz,1H),4.05–4.00(m,1H),3.99–3.96(m,2H),3.78–3.73(m,1H),3.32–3.29(m,2H),2.72(t,J＝6.5Hz,1H),2.61–2.55(m,1H),2.21–2.15(m,1H),2.12(dd,J＝13.7,3.6Hz,1H),1.90(t,J＝18.4Hz,3H),1.82–1.76(m,2H),1.71–1.66(m,1H).
步骤5：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((2-(3-(氟甲基)氮杂环丁烷-1-基)乙基)磺酰基)苯氧基)-6H-噻吩并[2,3-e]吲唑
Figure PCTCN2022134097-appb-000102
制备方法类似于实施例11步骤4-5的制备方法，不同的是将步骤4中的(E)-3-(4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-E]吲唑-3-基)氧基)苯基)丙-2-烯-1-醇替换为2-((4-((2-(2-(1,1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯基)磺酰基)乙烷-1-醇，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.57(s,1H),8.43(s,1H),7.80–7.76(m,2H),7.60–7.56(m,2H),7.50(dd,J＝9.8,2.8Hz,1H),7.36(td,J＝8.4,2.6Hz,1H),7.27(d,J＝8.9Hz,1H),7.16–7.12(m,2H),4.36(dd,J＝47.6,6.3Hz,2H),3.25(t,J＝7.0Hz,2H),3.04(t,J＝7.5Hz,2H),2.76–2.71(m,2H),2.58–2.51(m,3H),1.97(t,J＝19.0Hz,3H).MS(ESI-LR)m/z:604.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 26F 4N 3O 3S 2,604.1346；found 604.1369.
实施例14：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯 氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：3-(4-(苄氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000103
制备方法类似于实施例7步骤1至步骤2的制备方法，不同的是将4-(2-溴乙氧基)苯酚替换为4-苄氧基苯酚，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.20(d,J＝0.9Hz,1H),7.76(d,J＝8.9Hz,1H),7.59(dd,J＝9.0,0.9Hz,1H),7.47–7.32(m,5H),7.14–7.08(m,2H),7.01–6.93(m,2H),6.38(d,J＝0.6Hz,1H),5.79(dd,J＝9.3,2.7Hz,1H),5.06(s,2H),4.08–4.01(m,1H),3.84–3.72(m,1H),2.66–2.52(m,1H),2.22–2.09(m,2H),1.84–1.65(m,3H).
步骤2：4-((6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯酚的制备
Figure PCTCN2022134097-appb-000104
制备方法类似于实施例4步骤6的制备方法，不同的是将3-(4-(苄氧基)苯氧基)苯并[b]噻吩-6-羧酸甲酯替换为3-(4-(苄氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.20(d,J＝0.9Hz,1H),7.76(d,J＝8.9Hz,1H),7.59(dd,J＝9.0,0.9Hz,1H),7.08–7.02(m,2H),6.85–6.78(m,2H),6.35(s,1H),5.80(dd,J＝9.4,2.6Hz,1H),5.12(s,1H),4.06(dd,J＝11.6,3.0Hz,1H),3.83–3.73(m,1H),2.61(tdd,J＝13.4,10.0,4.0Hz,1H),2.23–2.06(m,2H),1.84–1.65(m,3H).
步骤3：2-溴-3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000105
制备方法类似于实施例4步骤7至步骤8的制备方法，不同的是将3-(4-羟基苯氧基)苯并[b]噻吩-6-羧酸甲酯替换为4-((6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯酚，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.16(d,J＝0.8Hz,1H),7.50(dd,J＝9.0,0.9Hz,1H),7.40(d,J＝9.0Hz,1H),6.91–6.85(m,2H),6.68–6.63(m,2H),5.75(dd,J＝9.3,2.7Hz,1H),4.80(tt,J＝6.4,4.1Hz,1H),4.30–4.21(m,2H),4.04–3.91(m,3H),3.78–3.68(m,1H),2.62–2.50(m,1H),2.20–2.06(m,2H),1.85–1.64(m,3H),1.44(s,9H).
步骤4：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000106
将上步得到的2-溴-3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(4.2g，7mmol)，2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷(2.6g，9.1mmol)，[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(1135mg，1.4mmol)和碳酸铯(4564mg，14mmol)溶于1,4-二氧六环(60mL)和水(6mL)的混合溶剂中，氩气保护，100℃加热反应18h。反应结束后，加水(100mL)，用乙酸乙酯(60mL×3)萃取，合并有机相，用饱和氯化钠溶液(100mL)洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-3/1)纯化后，浓缩得黄白色泡沫状固体2874mg，收率为60.4％。 1H NMR(400MHz,CDCl 3)δ8.20(d,J＝0.8Hz,1H),7.53(dd,J＝9.0,0.7Hz,1H),7.39(d,J＝9.0Hz,1H),7.36–7.30(m,2H),7.04(td,J＝8.1,2.7 Hz,1H),6.78–6.73(m,2H),6.57–6.52(m,2H),5.77(dd,J＝9.3,2.6Hz,1H),4.75(td,J＝6.1,3.0Hz,1H),4.22(dd,J＝9.7,6.4Hz,2H),4.08–3.99(m,1H),3.94(dd,J＝9.8,4.1Hz,2H),3.81–3.70(m,1H),2.58(q,J＝11.9,11.4Hz,1H),2.21–2.08(m,2H),1.87(t,J＝18.5Hz,3H),1.81–1.72(m,2H),1.70–1.66(m,1H),1.43(s,9H).
步骤5：3-(4-(氮杂环丁烷-3-氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)-6H-噻吩并[2,3-e]吲唑盐酸盐的制备
Figure PCTCN2022134097-appb-000107
将上步得到的2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(2.8g，4.12mmol)溶于异丙醇(50mL)，随后加入氯化氢的异丙醇溶液(41mL，4M)，50℃加热搅拌反应过夜，TLC(石油醚/乙酸乙酯＝3/1)监测反应完全应结束后，直接将反应液旋干，得粗品不经纯化，直接下一步。MS(ESI-LR)m/z:496.1(M+1) +.
步骤6：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000108
将上步得到的3-(4-(氮杂环丁烷-3-氧基)苯氧基)-2-(2-(1,1-二氟乙基)-4-氟苯基)-6H-噻吩并[2,3-e]吲唑盐酸盐(4mmol)悬浮于乙腈(50mL)中，加入碳酸钾(1382mg，10mmol)，氩气保护，室温搅拌20分钟。随后依次加入三乙胺(1.4mL，10mmol)，1-氟-3-碘丙烷(0.6mL，6mmol)，50℃反应过夜，TLC(二氯甲烷/甲醇＝10/1)监测反应并补加1-氟-3-碘丙烷和三乙胺直至反应完全。将反应液过滤除盐，乙酸乙酯(100mL)淋洗滤饼，滤液旋干拌样。经柱层析(乙酸乙酯/甲醇/二乙胺＝100/0.5/1-100/2/1)纯化后，浓缩得白色泡沫状固体1.5g，收率为67.5％。 1H NMR(500MHz,DMSO-d 6)δ13.52(s,1H),8.39(s,1H),7.57–7.51(m,2H),7.48(dd,J＝9.9,2.8Hz,1H),7.36(td,J＝8.3,2.8Hz,1H),7.27(d,J＝8.9Hz,1H),6.84–6.80(m,2H),6.73–6.64(m,2H),4.65(p,J＝5.6Hz,1H),4.44(dt,J＝47.5,6.1Hz,2H),3.66(td,J＝6.1,1.9Hz,2H),2.92–2.82(m,2H),2.55–2.39(m,2H),1.94(t,J＝19.0Hz,3H),1.64(dp,J＝25.7,6.5Hz,2H).MS(ESI-LR)m/z:556.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 26F 4N 3O 2S,556.1676；found 556.1688.
实施例15：(S)-2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)吡咯烷-3-基)氧基)苯 氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：(S)-3-(4-(1-Boc-3-羟基吡咯烷-3-氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000109
制备方法类似于实施例5步骤1的制备方法，不同的是将3-(4-羟基苯氧基)苯并[b]噻吩-6-羧酸甲酯替换为4-((6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯酚，制得标题化合物。 1H NMR(400MHz,CDCl 3)δ8.20(s,1H),7.74(d,J＝9.0Hz,1H),7.59(dd,J＝9.0,0.9Hz,1H),7.10(dd,J＝8.4,5.3Hz,2H),6.91–6.82(m,2H),6.40(s,1H),5.79(dd,J＝9.3,2.7Hz,1H),4.84(s,1H),4.07–4.01(m,1H),3.77(ddd,J＝13.5,8.6,3.0Hz,1H),3.63–3.47(m,4H),2.66–2.54(m,1H),2.23–2.15(m,2H),2.15–2.02(m,2H),1.84–1.74(m,2H),1.71–1.66(m,1H),1.47(s,9H).
步骤2：(S)-2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)吡咯烷-3-基)氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000110
制备方法类似于实施例14步骤3至步骤6的制备方法，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.52(s,1H),8.39(s,1H),7.57–7.51(m,2H),7.48(dd,J＝10.0,2.8Hz,1H),7.35(td,J＝8.3,2.7Hz,1H),7.28(d,J＝8.9Hz,1H),6.82(d,J＝8.7Hz,2H),6.75(d,J＝8.6Hz,2H),4.77(d,J＝6.7Hz,1H),4.47(dt,J＝47.4,6.0Hz,2H),2.94–2.54(m,6H),2.21(dq,J＝14.5,7.2Hz,1H),1.94(t,J＝18.9Hz,3H),1.87–1.72(m,3H).MS(ESI-LR)m/z:570.3(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 28F 4N 3O 2S,570.1833；found 570.1836.
实施例16：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)-2- 甲氧基苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000111
制备方法类似于实施例14的制备方法，不同的是将步骤1中的4-苄氧基苯酚替换为4(苄氧基)-2-甲氧基苯酚，制得标题化合物。 1H NMR(500MHz,DMSO-d 6)δ13.63–13.42(m,1H),8.37(s,1H),7.56(d,J＝8.9Hz,1H),7.45(dd,J＝8.8,5.6Hz,2H),7.38–7.26(m,2H),6.59(d,J＝8.8Hz,1H),6.49(d,J＝2.8Hz,1H),6.10(dd,J＝8.8,2.9Hz,1H),4.67(p,J＝5.5Hz,1H),4.44(dt,J＝47.4,6.1Hz,2H),3.71(s,3H),3.66(t,J＝7.0Hz,2H),2.93–2.84(m,2H),2.49(dd,J＝12.6,5.4Hz,2H),1.95(t,J＝19.0Hz,3H),1.65(dp,J＝25.8,6.6Hz,2H).MS(ESI-LR)m/z:586.1(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 28F 4N 3O 3S,586.1782；found 586.1786.
实施例17：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(3-氟-4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧 基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000112
制备方法类似于实施例14的制备方法，不同的是将步骤1中的4-苄氧基苯酚替换为4-(苄氧基)-3-氟苯酚，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.54(s,1H),8.40(s,1H),7.60–7.54(m,2H),7.49(dd,J＝9.9,2.8Hz,1H),7.38(td,J＝8.3,2.8Hz,1H),7.28(d,J＝8.9Hz,1H),6.91(dd,J＝12.2,3.0Hz,1H),6.82(t,J＝9.3Hz,1H),6.63(ddd,J＝9.0,3.0,1.4Hz,1H),4.74(p,J＝5.7Hz,1H),4.44(dt,J＝47.4,6.0Hz,2H),3.75(d,J＝7.2Hz,2H),3.08(d,J＝7.5Hz,2H),2.58(d,J＝7.2Hz,2H),1.95(t,J＝19.0Hz,3H),1.67(dp,J＝25.8,6.5Hz,2H).MS(ESI-LR)m/z:574.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 25F 5N 3O 2S,574.1582；found 574.1585.
实施例18：2-(4-氟-2-(1-氟环丙基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯氧 基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000113
制备方法类似于实施例14的制备方法，不同的是将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换2-(4-氟-2-(1-氟环丙基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.54(s,1H),8.40(s,1H),7.60–7.52(m,2H),7.46(dt,J＝9.5,2.2Hz,1H),7.32(td,J＝8.5,2.8Hz,1H),7.28(d,J＝8.9Hz,1H),6.87–6.83(m,2H),6.77–6.73(m,2H),4.87(p,J＝5.7Hz,1H),4.50(dt,J＝47.1,5.8Hz,2H),4.34–4.27(m,2H),3.85–3.74(m,2H),3.11(t,J＝7.6Hz,2H),1.85(dp,J＝26.4,6.2Hz,2H),1.28(dd,J＝18.3,7.1Hz,2H),1.10–1.05(m,2H).MS(ESI-LR)m/z:550.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 27F 3N 3O 2S,550.1771；found 550.1771.
实施例19：2-(4-氟-2-(1,1,1-三氟丙烷-2-基)苯基)-3-(4-(1-(3-氟丙基)氮杂环丁烷-3-基) 氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000114
制备方法类似于实施例14的制备方法，不同的是将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换2-(2-(1,1,1-三氟丙烷-2-基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(500MHz,DMSO-d 6)δ13.61–13.50(m,1H),8.42(s,1H),7.61–7.53(m,2H),7.40(dd,J＝10.2,2.7Hz,1H),7.33(d,J＝8.9Hz,1H),7.27(td,J＝8.4,2.7Hz,1H),6.80–6.75(m,2H),6.71–6.65(m,2H),4.64(p,J＝5.6 Hz,1H),4.43(dt,J＝47.4,6.0Hz,2H),3.93(p,J＝8.4Hz,1H),3.66(dd,J＝8.2,5.8Hz,2H),2.89(t,J＝6.7Hz,2H),2.48(t,J＝7.0Hz,2H),1.63(dp,J＝25.7,6.5Hz,2H),1.40(d,J＝7.1Hz,3H).MS(ESI-LR)m/z:588.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 227F 5N 3O 2S,588.1739；found 588.1741.
实施例20：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基)苯 氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氨基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑
Figure PCTCN2022134097-appb-000115
将实施例13中步骤1所得3-(4-碘苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(320mg，0.67mmol)，1-叔丁氧羰基-3-氨基氮杂环丁烷(172mg，1mmol)，甲烷磺酸(2-二环己基膦)-3,6-二甲氧基-2',4',6'-三异丙基-1,1'-联苯)(2'-氨基-1,1'-联苯-2-基)钯(33mg，0.036mmol)，碳酸铯(429mg，1.32mmol)溶于1,4-二氧六环(5mL)，氩气保护，微波100℃反应6h，反应结束后向反应液中加入水(15mL)，用乙酸乙酯(15mL×3)萃取，合并有机相，用饱和氯化钠溶液(20mL)洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-3/1)纯化后，浓缩得土黄色粉末228mg，收率为65.2％。 1H NMR(400MHz,DMSO-d 6)δ8.39(s,1H),7.78(d,J＝9.0Hz,1H),7.72(d,J＝8.9Hz,1H),7.05–6.98(m,2H),6.58–6.52(m,3H),6.25(d,J＝6.1Hz,1H),5.94(dd,J＝9.6,2.4Hz,1H),4.24–4.08(m,3H),3.90(d,J＝11.6Hz,1H),3.83–3.72(m,1H),3.67–3.58(m,2H),2.47–2.38(m,1H),2.09–1.96(m,2H),1.85–1.69(m,1H),1.64–1.54(m,2H),1.38(s,9H).
步骤2：3-(4-((1-叔丁氧羰基氮杂环丁烷-3-基)-N-叔丁氧羰基氨基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000116
将3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氨基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(600mg，1.15mmol)溶于甲醇(15mL)，随后加入二碳酸二叔丁酯(26.5mL，115.3mmol)，65℃加热回流过夜。反应结束，加水(30mL)，用乙酸乙酯(20mL×3)萃取，合并有机相，用饱和氯化钠溶液(20mL)洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-3/1)纯化后，浓缩得黄色油状物578mg，收率为80.9％。 1H NMR(400MHz,CDCl 3)δ8.21(d,J＝0.8Hz,1H),7.65(d,J＝8.9Hz,1H),7.57(dd,J＝9.0,0.9Hz,1H),7.11–7.04(m,4H),6.66(s,1H),5.79(dd,J＝9.4,2.7Hz,1H),4.82–4.71(m,1H),4.09–4.02(m,3H),3.85–3.72(m,3H),2.59(q,J＝12.5,11.2Hz,1H),2.19–2.06(m, 2H),1.84–1.68(m,3H),1.60(s,9H),1.41(s,9H).
步骤3：2-溴-3-(4-((1-叔丁氧羰基氮杂环丁烷-3-基)-N-叔丁氧羰基氨基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000117
将上步得到的3-(4-((1-叔丁氧羰基氮杂环丁烷-3-基)-N-叔丁氧羰基氨基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(75mg，0.12mmol)溶于乙腈(5mL)中，随后加入N-溴代丁二酰亚胺(24mg，0.13mmol)，氩气保护。避光条件下，室温搅拌过夜。饱和硫代硫酸钠溶液(10mL)淬灭反应，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和氯化钠溶液(10mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-5/1)纯化后，浓缩得无色油状物75mg，收率89.3％。 1H NMR(400MHz,CDCl 3)δ8.17(s,1H),7.52(d,J＝9.0Hz,1H),7.41(d,J＝9.0Hz,1H),7.06–6.98(m,2H),6.97–6.89(m,2H),5.76(dd,J＝9.3,2.6Hz,1H),4.75(s,1H),4.04(q,J＝8.9Hz,3H),3.83–3.72(m,3H),2.57(q,J＝9.3,8.5Hz,1H),2.25–2.05(m,2H),1.82–1.68(m,3H),1.41(s,18H).
步骤4：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-叔丁氧羰基氮杂环丁烷-3-基)-N-叔丁氧羰基氨基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000118
将上步得到的2-溴-3-(4-((1-叔丁氧羰基氮杂环丁烷-3-基)-N-叔丁氧羰基氨基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(158mg，0.226mmol)，2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷(123mg，0.43mmol)，三(二亚苄基丙酮)二钯(20mg，0.022mmol)，2-二环己基膦-2',4',6'-三异丙基联苯(20mg，0.042mmol)，磷酸钾(137mg，0.65mmol)溶于1,4-二氧六环(5mL)和水(0.5mL)的混合溶剂中，110℃微波反应2h。反应结束后，直接将反应液旋干硅胶拌样，经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得粉白色泡沫状固体126mg，收率为56.6％。 1H NMR(400MHz,CDCl 3)δ8.24(d,J＝0.8Hz,1H),7.57(dd,J＝9.0,0.9Hz,1H),7.44(d,J＝8.9Hz,1H),7.40–7.32(m,2H),7.07(td,J＝8.1,2.7Hz,1H),6.93(d,J＝8.8Hz,2H),6.85–6.81(m,2H),5.81(dd,J＝9.3,2.7Hz,1H),4.75(s,1H),4.07–4.02(m,3H),3.75(s,3H),2.66–2.56(m,1H),2.20–2.09(m,2H),1.90(t,J＝18.5Hz,3H),1.82–1.71(m,3H),1.42(s,18H).
步骤5：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000119
制备方法类似于实施例14中步骤5-6的制备方法，不同的是将步骤5中的2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑替换为2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-叔丁氧羰基氮杂环丁烷-3-基)-N-叔丁氧羰基氨基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.48(s,1H),8.36(s,1H),7.56–7.50(m,2H),7.48(dd,J＝9.9,2.8Hz,1H),7.37(td,J＝8.3,2.8Hz,1H),7.27(d,J＝8.9Hz,1H),6.67(d,J＝8.5Hz,2H),6.36(d,J＝8.7Hz,2H),5.76(d,J＝7.2Hz,1H),4.44(dt,J＝47.4,6.1Hz,2H),3.81(h,J＝6.6Hz,1H),3.58(t,J＝6.8Hz,2H),2.68(t,J＝6.7Hz,2H),2.44(t,J＝7.0Hz,2H),1.93(t,J＝18.9Hz,3H),1.63(dp,J＝25.7,6.5Hz,2H).MS(ESI-LR)m/z:555.3(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 27F 4N 4OS,555.1836；found 555.1846.
实施例21：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基)-6- 氟苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000120
制备方法类似于实施例20的制备方法，不同的是将4-碘苯酚替换为2-氟-4-碘苯酚制得标题化合物。 1H NMR(500MHz,Methanol-d 4)δ8.24(s,1H),7.56(d,J＝8.9Hz,1H),7.53(d,J＝9.0Hz,1H),7.34–7.26(m,2H),7.06(td,J＝8.3,2.8Hz,1H),6.60(t,J＝9.0Hz,1H),6.27(dd,J＝13.1,2.7Hz,1H),6.08(dd,J＝8.9,2.7Hz,1H),4.49(dt,J＝47.3,5.7Hz,2H),4.09(p,J＝6.3Hz,1H),4.02(t,J＝8.0Hz,2H),3.31(t,J＝7.5Hz,2H),2.93(t,J＝7.5Hz,2H),1.90–1.78(m,5H).MS(ESI-LR)m/z:573.3(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 26F 5N 4OS,573.1742；found 573.1759.
实施例22：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基)-6- 氟苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000121
制备方法类似于实施例20的制备方法，不同的是将4-碘苯酚替换为2-三氟甲基-4-碘苯酚制得标题化合物。 1H NMR(500MHz,DMSO-d 6)δ13.71–13.45(s,1H),8.40(s,1H),7.58(ddd,J＝12.4,8.7,4.7Hz,2H),7.48(dd,J＝9.8,2.8Hz,1H),7.37(t,J＝8.5Hz,1H),7.18 (dd,J＝9.0,5.3Hz,1H),6.79(t,J＝3.9Hz,1H),6.69(dd,J＝8.9,3.1Hz,1H),6.56(dt,J＝7.2,3.3Hz,1H),6.32(t,J＝6.4Hz,1H),4.45(dt,J＝47.4,6.0Hz,2H),3.98(p,J＝6.6Hz,1H),3.75(t,J＝7.2Hz,2H),2.99–2.89(m,2H),2.62(t,J＝7.1Hz,2H),1.94(t,J＝18.9Hz,3H),1.69(dp,J＝26.0,6.7Hz,2H).MS(ESI-LR)m/z:623.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 26F 7N 4OS,623.171；found 623.172.
实施例23：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基)-6- 甲氧基苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000122
制备方法类似于实施例20的制备方法，不同的是将4-碘苯酚替换为2-甲氧基-4-碘苯酚制得标题化合物。 1H NMR(600MHz,Methanol-d 4)δ8.22(s,1H),7.56(d,J＝8.9Hz,1H),7.48(d,J＝8.9Hz,1H),7.29(dd,J＝9.8,2.7Hz,1H),7.24(dd,J＝8.5,5.7Hz,1H),7.01(td,J＝8.2,2.7Hz,1H),6.48(d,J＝8.6Hz,1H),6.14(d,J＝2.5Hz,1H),5.82(dd,J＝8.7,2.5Hz,1H),4.45(dt,J＝47.3,5.8Hz,2H),3.99(p,J＝6.7Hz,1H),3.78(t,J＝7.4Hz,2H),3.62(s,3H),2.98(t,J＝7.4Hz,2H),2.68(t,J＝7.5Hz,2H),1.85(t,J＝18.7Hz,3H),1.75(dt,J＝25.9,6.8Hz,2H). 19F NMR(471MHz,MeOD)δ-84.22,-113.70(q,J＝8.2Hz),-221.38(tt,J＝48.9,26.0Hz).MS(ESI-LR)m/z:585.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 29F 4N 4O 2S,585.1942；found 585.1935.
实施例24：(S)-2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)吡咯烷-3-基)氨基)苯 氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000123
制备方法类似于实施例20的制备方法，不同的是将步骤1中的1-叔丁氧羰基-3-氨基氮杂环丁烷替换为(R)-3-氨基吡咯烷-1-羧酸叔丁酯制得标题化合物。 1H NMR(500MHz,DMSO-d 6)δ13.52(s,1H),8.37(s,1H),7.54(dt,J＝9.0,4.4Hz,2H),7.48(dd,J＝9.9,2.8Hz,1H),7.37(d,J＝8.5Hz,1H),7.30(dt,J＝8.9,2.5Hz,1H),6.69(d,J＝8.3Hz,2H),6.42(d,J＝8.3Hz,2H),5.52–5.47(m,1H),4.48(dt,J＝47.3,5.9Hz,2H),3.84–3.79(m,1H),3.01–2.92(m,1H),2.85–2.76(m,1H),2.72–2.60(m,3H),2.55–2.51(m,1H),2.16(dq,J＝14.5,7.3Hz,1H),1.93(t,J＝19.0Hz,3H),1.89–1.79(m,2H),1.60(dq,J＝12.8,6.3Hz,1H).MS(ESI-LR)m/z:569.1(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 29F 4N 4OS,569.1993；found 569.1933.
实施例25：2-(2-(4-氟-2-(2,2,2-三氟乙基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨 基)-苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000124
制备方法类似于实施例20的制备方法，不同的是将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(4-氟-2-(2,2,2-三氟乙基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.54(s,1H),8.40(s,1H),7.57–7.53(m,2H),7.38–7.33(m,2H),7.28(td,J＝8.4,2.8Hz,1H),6.67–6.63(m,2H),6.37–6.32(m,2H),5.76(d,J＝7.1Hz,1H),4.43(dt,J＝47.4,6.1Hz,2H),3.84–3.76(m,3H),3.59(td,J＝6.3,1.7Hz,2H),2.70(t,J＝6.8Hz,2H),2.45(t,J＝7.0Hz,2H),1.63(dp,J＝25.7,6.5Hz,2H). 19F NMR(471MHz,DMSO)δ-63.25(t,J＝11.2Hz),-112.59(q,J＝8.5,8.1Hz),-217.95–-218.33(m).MS(ESI-LR)m/z:573.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 25F 5N 4OS,573.3822；found 573.3833.
实施例26：2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨基) 苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000125
制备方法类似于实施例20的制备方法，不同的是将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(500MHz,DMSO-d 6)δ13.58(s,1H),8.37(s,1H),7.73(dd,J＝11.6,8.0Hz,1H),7.63(dd,J＝10.8,7.9Hz,1H),7.55(d,J＝8.9Hz,1H),7.29(d,J＝8.9Hz,1H),6.76(d,J＝8.5Hz,2H),6.44(d,J＝8.5Hz,2H),6.25(d,J＝7.0Hz,1H),4.53(dt,J＝47.1,5.7Hz,2H),4.42–4.15(m,3H),3.90–3.78(m,2H),3.29(t,J＝7.6Hz,2H),2.00–1.83(m,5H). 19F NMR(471MHz,DMSO)δ-80.60(q,J＝19.1Hz),-136.00(dt,J＝20.7,9.9Hz),-136.34(dt,J＝20.9,9.6Hz),-219.24(tt,J＝48.5,26.5Hz).MS(ESI-LR)m/z:573.3(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 26F 5N 4OS,573.1742；found 573.1742.
实施例27：2-((2-(4-氟-2-(2-氟丙烷-2-基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨 基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000126
制备方法类似于实施例20的制备方法，不同的是将步骤4中的2-(2-(1,1-二氟乙基)-4- 氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-((2-(4-氟-2-(2-氟丙烷-2-基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(500MHz,Methanol-d 4)δ8.23(s,1H),7.46(d,J＝8.8Hz,1H),7.40(d,J＝9.0Hz,1H),7.30(dd,J＝10.9,2.8Hz,1H),7.21(dd,J＝8.5,5.9Hz,1H),6.88(td,J＝8.1,2.8Hz,1H),6.67–6.62(m,2H),6.44–6.39(m,2H),4.46(dt,J＝47.3,5.8Hz,2H),4.05(p,J＝6.6Hz,1H),3.90(td,J＝7.0,1.8Hz,2H),3.19–3.13(m,2H),2.80(t,J＝7.4Hz,2H),1.85–1.72(m,2H),1.63(d,J＝23.0Hz,6H). 19F NMR(471MHz,Methanol-d 4)δ-113.91(dt,J＝12.5,6.7Hz),-127.64(h,J＝22.8Hz),-221.46(tt,J＝47.5,26.3Hz).MS(ESI-LR)m/z:551.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 30F 3N 4OS,551.2087；found 551.2097.
实施例28：2-((2-(4-氟-2-(1,1,1,2-四氟丙烷-2-基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3- 基)氨基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000127
制备方法类似于实施例20的制备方法，不同的是将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-((2-(4-氟-2-(1,1,1,2-四氟丙烷-2-基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.48(s,1H),8.36(s,1H),7.52(dd,J＝8.7,5.8Hz,2H),7.48(d,J＝10.5Hz,1H),7.36(td,J＝8.2,2.7Hz,1H),7.28(d,J＝8.9Hz,1H),6.65(d,J＝8.4Hz,2H),6.36(d,J＝8.6Hz,2H),5.75(d,J＝7.2Hz,1H),4.44(dt,J＝47.4,6.0Hz,2H),3.83(h,J＝6.6Hz,1H),3.60(t,J＝6.8Hz,2H),2.71(t,J＝6.7Hz,2H),2.47(t,J＝7.0Hz,2H),1.88(d,J＝24.2Hz,3H),1.64(dp,J＝25.8,6.5Hz,2H).MS(ESI-LR)m/z:605.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 27F 6N 4OS,605.1804；found 605.1801.
实施例29：2-((2-(4-氟-2-(1,1,1-三氟-2-甲基丙烷-2-基)苯基)-3-(4-((1-(3-氟丙基)氮杂环 丁烷-3-基)氨基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000128
制备方法类似于实施例20的制备方法，不同的是将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-((2-(4-氟-2-(1,1,1-三氟-2-甲基丙烷-2-基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(500MHz,DMSO-d 6)δ13.52(s,1H),8.37(s,1H),7.60–7.46(m,2H),7.38(d,J＝9.1Hz,1H),7.29–7.13(m,2H),6.69(d,J＝8.3Hz,2H),6.38(d,J＝8.4Hz,2H),5.89(s,1H),4.60–4.34(m,2H),3.88(d,J＝57.5Hz,3H),3.13–2.93(m,2H),2.81–2.60(m,2H),1.72(d,J＝25.3Hz,2H), 1.58(s,3H),1.41(s,3H).MS(ESI-LR)m/z:601.3(M+1) +.HRMS(ESI):Anal.Calcd for C 31H 30F 5N 4OS,601.2055；found 601.205.
实施例30：2-(2-(4-氟-2-(2,2,2-三氟乙基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨 基)-6-氟苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000129
制备方法类似于实施例20的制备方法，不同的是将4-碘苯酚替换为2-氟-4-碘苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(4-氟-2-(2,2,2-三氟乙基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(500MHz,DMSO-d 6)δ13.54(s,1H),8.40(s,1H),7.61(d,J＝8.9Hz,1H),7.50–7.44(m,2H),7.30(dd,J＝9.9,2.7Hz,1H),7.24(td,J＝8.5,2.8Hz,1H),6.59(t,J＝9.2Hz,1H),6.28(dd,J＝13.6,2.7Hz,1H),6.10–6.01(m,2H),4.43(dt,J＝47.4,6.0Hz,2H),3.82(q,J＝6.6Hz,1H),3.73(q,J＝11.1Hz,2H),3.58(t,J＝6.7Hz,2H),2.70(t,J＝6.7Hz,2H),2.45(t,J＝7.0Hz,2H),1.63(dq,J＝25.7,6.5Hz,2H).MS(ESI-LR)m/z:591.3(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 25F 6N 4OS,591.1648；found 591.1661.
实施例31：2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨 基)-6-氟苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000130
制备方法类似于实施例20的制备方法，不同的是将4-碘苯酚替换为2-氟-4-碘苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(400MHz,DMSO-d 6)δ13.55(s,1H),8.38(s,1H),7.70(dd,J＝11.7,8.1Hz,1H),7.59(dd,J＝8.8,1.0Hz,1H),7.52(dd,J＝10.9,7.9Hz,1H),7.43(d,J＝8.9Hz,1H),6.66(t,J＝9.2Hz,1H),6.29(dd,J＝13.7,2.7Hz,1H),6.14(d,J＝7.0Hz,1H),6.10–6.03(m,1H),4.44(dt,J＝47.4,6.1Hz,2H),3.83(h,J＝6.5Hz,1H),3.58(td,J＝6.3,1.7Hz,2H),2.71–2.61(m,2H),2.45(t,J＝7.0Hz,2H),1.89(t,J＝19.0Hz,3H),1.64(dp,J＝25.6,6.6Hz,2H). 19F NMR(471MHz,DMSO-d 6)δ-80.84(q,J＝19.0Hz),-132.08(t,J＝11.9Hz),-136.10(dt,J＝20.5,9.7Hz),-136.52(dt,J＝21.1,9.7Hz),-218.14(tt,J＝48.4,25.1Hz).MS(ESI-LR)m/z:591.3(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 25F 6N 4OS,591.1648；found 591.1653.
实施例32：2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨 基)-6-甲氧基苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000131
制备方法类似于实施例20的制备方法，不同的是将4-碘苯酚替换为2-甲氧基-4-碘苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(500MHz,DMSO-d 6)δ13.52(s,1H),8.34(s,1H),7.64(dd,J＝11.6,8.1Hz,1H),7.56(d,J＝8.8Hz,1H),7.43(d,J＝8.9Hz,1H),7.38(dd,J＝10.8,7.9Hz,1H),6.52(d,J＝8.6Hz,1H),6.13(d,J＝2.6Hz,1H),5.83–5.77(m,2H),4.44(dt,J＝47.4,6.1Hz,2H),3.85(h,J＝6.6Hz,1H),3.63(s,3H),3.59(t,J＝6.7Hz,2H),2.69(t,J＝6.6Hz,2H),2.45(t,J＝6.9Hz,2H),1.89(t,J＝19.0Hz,3H),1.64(dp,J＝25.7,6.5Hz,2H). 19F NMR(471MHz,DMSO-d 6)δ-80.49(p,J＝26.1,19.3Hz),-136.53(dt,J＝21.2,10.0Hz),-136.72(dt,J＝21.3,9.7Hz),-218.10(tt,J＝48.0,25.3Hz).MS(ESI-LR)m/z:603.3(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 28F 5N 4O 2S,603.1848；found 603.1851.
实施例33：2-(2-(4-氟-2-(2,2,2-三氟乙基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨 基)-6-甲氧基苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000132
制备方法类似于实施例20的制备方法，不同的是将4-碘苯酚替换为2-甲氧基-4-碘苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(4-氟-2-(2,2,2-三氟乙基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.55(s,1H),8.37(s,1H),7.57(dd,J＝9.0,2.6Hz,1H),7.47–7.39(m,2H),7.27(dd,J＝9.8,2.8Hz,1H),7.22(td,J＝8.4,2.8Hz,1H),6.45(dd,J＝8.7,2.5Hz,1H),6.12(t,J＝2.6Hz,1H),5.88(d,J＝7.3Hz,1H),5.79(dd,J＝8.7,2.6Hz,1H),4.46(dt,J＝47.3,6.0Hz,2H),3.91(q,J＝6.7Hz,1H),3.77–3.70(m,4H),3.68(s,3H),2.94–2.81(m,2H),2.60(t,J＝7.1Hz,2H),1.69(dp,J＝26.0,6.6Hz,2H).MS(ESI-LR)m/z:603.3(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 28F 5N 4O 2S,603.1848；found 603.1861.
实施例34：2-(2-(4-氟-2-(2-氟丙烷-2-基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氨 基)-6-甲氧基苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000133
制备方法类似于实施例20的制备方法，不同的是将4-碘苯酚替换为2-甲氧基-4-碘苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-((2-(4-氟-2-(2-氟丙烷-2-基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷得标题化合物。 1H NMR(500MHz,Methanol-d 4)δ8.21(d,J＝2.4Hz,1H),7.52(dd,J＝8.9,2.4Hz,1H),7.47(dd,J＝9.1,3.4Hz,1H),7.26(dt,J＝11.0,2.8Hz,1H),7.08(dd,J＝8.6,5.5Hz,1H),6.81(tdd,J＝8.1,5.1,2.8Hz,1H),6.46(dd,J＝8.6,2.2Hz,1H),6.17(d,J＝2.5Hz,1H),5.83(dd,J＝8.6,2.9Hz,1H),4.45(dtd,J＝47.2,5.9,2.7Hz,2H),4.03(td,J＝6.7,2.7Hz,1H),3.86(q,J＝7.8,6.1Hz,2H),3.61(s,3H),3.14–3.06(m,2H),2.76(td,J＝7.7,4.0Hz,2H),1.85–1.71(m,2H),1.61(d,J＝23.2,6H). 19F NMR(471MHz,Methanol-d 4)δ-113.97(dtd,J＝34.3,17.8,8.7Hz),-126.77–-127.41(m),-220.88–-221.38(m).MS(ESI-LR)m/z:581.3(M+1) +.HRMS(ESI):Anal.Calcd for C 31H 32F 3N 4O 2S,581.2193；found 581.2193.
实施例35：2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧 基)-6-甲氧基苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000134
制备方法类似于实施例14的制备方法，不同的是将步骤1中的4-苄氧基苯酚替换为2-甲氧基-4-苄氧基苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-((2-(1,1-二氟乙基)-4,5-二氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷得标题化合物。 1H NMR(400MHz,DMSO-d 6)δ13.51(s,1H),8.36(s,1H),7.69(dd,J＝11.6,8.1Hz,1H),7.57(d,J＝8.9Hz,1H),7.48(dd,J＝10.9,8.0Hz,1H),7.36(d,J＝8.9Hz,1H),6.65(d,J＝8.8Hz,1H),6.49(d,J＝2.8Hz,1H),6.10(dd,J＝8.9,2.8Hz,1H),4.70(p,J＝5.7Hz,1H),4.45(dt,J＝47.4,6.0Hz,2H),3.77–3.65(m,5H),3.02–2.88(m,2H),2.60–2.51(m,2H),1.92(t,J＝19.0Hz,3H),1.67(dp,J＝25.6,6.5Hz,2H). 19F NMR(471MHz,DMSO-d 6)δ-73.50,-80.63(q,J＝19.4Hz),-135.93–-136.73(m),-218.29(tt,J＝47.9,25.3Hz).MS(ESI-LR)m/z:604.3(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 26F 5N 3O 3S,604.379；found 604.3796.
实施例36：2-(2-(4-氟-2-(2,2,2-三氟乙基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧 基)-6-氟苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000135
制备方法类似于实施例14的制备方法，不同的是将步骤1中的4-苄氧基苯酚替换为2-氟-4-苄氧基苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧 硼杂环戊烷替换为2-((2-(4-氟-2-(2,2,2-三氟乙基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷得标题化合物。 1H NMR(400MHz,DMSO-d 6)δ13.55(s,1H),8.42(s,1H),7.62(d,J＝8.9Hz,1H),7.52–7.42(m,2H),7.32(dd,J＝9.9,2.6Hz,1H),7.26(td,J＝8.5,2.7Hz,1H),6.79–6.71(m,2H),6.39(dd,J＝8.8,2.9Hz,1H),4.68(q,J＝5.7Hz,1H),4.44(dt,J＝47.4,6.0Hz,2H),3.74(q,J＝11.2Hz,2H),3.68–3.58(m,2H),2.87(t,J＝6.6Hz,2H),2.50–2.43(m,2H),1.64(dp,J＝25.4,6.4Hz,2H). 19F NMR(471MHz,DMSO-d 6)δ-62.85–-63.63(m),-112.12,-130.49,-218.12(tt,J＝49.5,25.6Hz).MS(ESI-LR)m/z:592.3(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 24F 6N 3O 2S,592.1488；found 592.1493.
实施例37：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)-6- 氟苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000136
制备方法类似于实施例14的制备方法，不同的是将步骤1中的4-苄氧基苯酚替换为2-氟-4-苄氧基苯酚，制得标题化合物。 1H NMR(400MHz,DMSO-d 6)δ13.54(s,1H),8.39(s,1H),7.59(d,J＝8.9Hz,1H),7.51–7.44(m,2H),7.37(d,J＝8.8Hz,1H),7.33(dd,J＝8.4,2.8Hz,1H),6.83–6.72(m,2H),6.43(dd,J＝8.9,3.6Hz,1H),4.71(p,J＝5.7Hz,1H),4.45(dt,J＝47.4,6.0Hz,2H),3.84–3.69(m,2H),3.11–2.92(m,2H),2.63–2.54(m,2H),1.92(t,J＝19.0Hz,3H),1.67(dq,J＝25.6,6.5Hz,2H). 19F NMR(471MHz,Methanol-d 4)δ-84.73,-112.94(d,J＝25.5Hz),-131.64(q,J＝17.0,11.1Hz),-221.40(tq,J＝48.8,27.7,25.4Hz).MS(ESI-LR)m/z:574.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 25F 5N 3O 2S,574.1582；found 574.1592.
实施例38：2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧 基)-6-氟苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000137
制备方法类似于实施例14的制备方法，不同的是将步骤1中的4-苄氧基苯酚替换为2-氟-4-苄氧基苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.55(s,1H),8.41(s,1H),7.72(dd,J＝11.6,7.9Hz,1H),7.63–7.54(m,2H),7.41(d,J＝8.8Hz,1H),6.87–6.76(m,2H),6.44(d,J＝9.1Hz,1H),4.70(p,J＝5.5Hz,1H),4.44(dt,J＝47.4,6.1Hz,2H),3.66(t,J＝6.9Hz,2H),2.87(t,J＝6.5Hz,2H),2.49–2.46(m,2H),1.91(t,J＝18.9Hz,3H),1.65(dq,J＝25.4,6.6Hz, 2H). 19F NMR(471MHz,DMSO-d 6)δ-80.73,-130.23,-135.91(d,J＝250.1Hz),-217.68–-218.69(m).MS(ESI-LR)m/z:592.2(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 24F 6N 3O 2S,592.1488；found 592.1494.
实施例39：2-(2-(4-氟-2-(2-氟丙烷-2-基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧 基)-6-氟苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000138
制备方法类似于实施例14的制备方法，不同的是将步骤1中的4-苄氧基苯酚替换为2-氟-4-苄氧基苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.52(s,1H),8.37(s,1H),7.58(d,J＝8.9Hz,1H),7.39–7.30(m,3H),7.12(td,J＝8.2,2.8Hz,1H),6.81–6.72(m,2H),6.43(dd,J＝9.0,2.9Hz,1H),4.69(p,J＝5.7Hz,1H),4.44(dt,J＝47.4,6.1Hz,2H),3.68(t,J＝7.0Hz,2H),2.92(t,J＝6.7Hz,2H),2.53–2.50(m,2H),1.70–1.57(m,8H).MS(ESI-LR)m/z:570.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 28F 4N 3O 2S,570.1833；found 570.1829.
实施例40：2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧 基)-6-氟苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000139
制备方法类似于实施例14的制备方法，不同的是将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(1,1-二氟乙基)-4,5-二氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.53(s,1H),8.39(s,1H),7.73(dd,J＝11.5,8.0Hz,1H),7.68–7.59(m,1H),7.55(d,J＝8.9Hz,1H),7.29(d,J＝8.9Hz,1H),6.85(d,J＝8.5Hz,2H),6.69(d,J＝8.6Hz,2H),4.66(p,J＝5.8Hz,1H),4.44(dt,J＝47.4,6.1Hz,2H),3.67(t,J＝6.8Hz,2H),2.89(t,J＝6.6Hz,2H),2.50–2.46(m,2H),1.93(t,J＝18.9Hz,3H),1.64(dp,J＝25.9,6.6Hz,2H).MS(ESI-LR)m/z:574.3(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 25F 5N 3O 2S,574.1582；found 574.1577.
实施例41：2-(2-(4-氟-2-(2,2,2-三氟乙基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧 基)-6-甲氧基苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000140
制备方法类似于实施例14的制备方法，不同的是将步骤1中的4-苄氧基苯酚替换为2-甲氧基-4-苄氧基苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(4-氟-2-(2,2,2-三氟乙基)苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.51(s,1H),8.38(s,1H),7.58(d,J＝8.9Hz,1H),7.46–7.40(m,2H),7.27(dd,J＝9.8,2.8Hz,1H),7.23(td,J＝8.5,2.8Hz,1H),6.57(d,J＝8.8Hz,1H),6.43(d,J＝2.8Hz,1H),6.07(dd,J＝8.8,2.8Hz,1H),4.66(p,J＝5.7Hz,1H),4.44(dt,J＝47.4,6.1Hz,2H),3.77–3.69(m,5H),3.68–3.63(m,2H),2.85(t,J＝6.8Hz,2H),2.50–2.46(m,2H),1.65(dp,J＝25.6,6.5Hz,2H).MS(ESI-LR)m/z:604.2(M+1) +.HRMS(ESI):Anal.Calcd for C 30H 27F 5N 3O 3S,604.1688；found 604.1695.
实施例42：2-(2-(4-氟-2-(2-氟丙烷-2-基)苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧 基)-6-甲氧基苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000141
制备方法类似于实施例14的制备方法，不同的是将步骤1中的4-苄氧基苯酚替换为2-甲氧基-4-苄氧基苯酚，将步骤4中的2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷替换为2-(2-(4-氟-2-(2-氟丙烷-2-基)苯基基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.49(s,1H),8.35(s,1H),7.54(d,J＝8.9Hz,1H),7.36–7.27(m,3H),7.11(td,J＝8.2,2.8Hz,1H),6.58(d,J＝8.8Hz,1H),6.50(d,J＝2.8Hz,1H),6.10(dd,J＝8.9,2.8Hz,1H),4.67(p,J＝5.7Hz,1H),4.44(dt,J＝47.4,6.1Hz,2H),3.71(s,3H),3.70–3.66(m,2H),2.90(t,J＝6.7Hz,2H),2.52–2.49(m,2H),1.70–1.58(m,8H).MS(ESI-LR)m/z:582.3(M+1) +.HRMS(ESI):Anal.Calcd for C 31H 31F 3N 3O 3S,582.2033；found 582.2033.
实施例43： 2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯 氧基)-6H-噻吩并[2,3-e]吲唑L-酒石酸盐的制备
Figure PCTCN2022134097-appb-000142
2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑(55mg，0.1mmol)溶于乙腈(1.5mL)，随后滴加L-酒石酸(15mg，0.1mmol)的水(0.5mL)溶液，室温搅拌30min，随后冻干。将冻干粉末悬浮于甲基叔丁基醚(2mL)， 室温搅拌30min，过滤，滤饼35℃真空干燥得白色粉末56mg，收率80％。 1H NMR(600MHz,DMSO-d 6)δ13.50(s,1H),8.39(s,1H),7.56–7.52(m,2H),7.49(dd,J＝9.9,2.8Hz,1H),7.37(td,J＝8.3,2.8Hz,1H),7.25(d,J＝8.8Hz,1H),6.84–6.80(m,2H),6.71–6.68(m,2H),4.69(q,J＝5.7Hz,1H),4.45(dt,J＝47.4,6.0Hz,2H),4.20(s,2H),3.78(t,J＝7.2Hz,2H),3.09–3.00(m,2H),2.60(t,J＝7.1Hz,2H),1.94(t,J＝19.0Hz,3H),1.67(dq,J＝25.7,6.5Hz,2H).
实施例44： 2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯 氧基)-6H-噻吩并[2,3-e]吲唑甲磺酸盐的制备
Figure PCTCN2022134097-appb-000143
制备方法类似于实施例43中的制备方法，不同的是将L-酒石酸的水溶液替换为甲磺酸的乙腈溶液，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.52(s,1H),9.93(s,1H),8.39(s,1H),7.57–7.53(m,2H),7.50(dd,J＝9.9,2.8Hz,1H),7.38(td,J＝8.3,2.8Hz,1H),7.25(d,J＝8.9Hz,1H),6.89–6.85(m,2H),6.76–6.72(m,2H),4.97–4.85(m,1H),4.56–4.32(m,4H),4.07–3.88(m,2H),3.27–3.16(m,2H),2.31(s,3H),1.95(t,J＝19.0Hz,3H),1.86(dt,J＝26.7,7.0Hz,2H).
实施例45： 2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯 氧基)-6H-噻吩并[2,3-e]吲唑马来酸盐的制备
Figure PCTCN2022134097-appb-000144
制备方法类似于实施例43中的制备方法，不同的是将L-酒石酸替换为马来酸，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.51(s,1H),8.40(s,1H),7.57–7.52(m,2H),7.50(dd,J＝9.9,2.8Hz,1H),7.38(td,J＝8.3,2.8Hz,1H),7.25(d,J＝8.9Hz,1H),6.87(d,J＝8.6Hz,2H),6.74(d,J＝8.6Hz,2H),6.03(s,2H),4.96–4.84(m,1H),4.57–4.36(m,4H),4.14–3.88(m,2H),3.28–3.17(m,2H),2.00–1.80(m,5H).
实施例46： 2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯 氧基)-6H-噻吩并[2,3-e]吲唑柠檬盐的制备
Figure PCTCN2022134097-appb-000145
制备方法类似于实施例43中的制备方法，不同的是将L-酒石酸替换为柠檬酸，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.50(s,1H),8.39(s,1H),7.57–7.52(m,2H),7.49(dd,J＝9.9,2.8Hz,1H),7.37(td,J＝8.3,2.8Hz,1H),7.25(d,J＝8.9Hz,1H),6.86–6.81(m,2H),6.73–6.68(m,2H),4.76(p,J＝5.7Hz,1H),4.46(dt,J＝47.2,5.9Hz,2H),4.05–3.91(m,2H),3.40–3.30(m,2H),2.85–2.73(m,2H),2.66(d,J＝15.3Hz,2H),2.58(d,J＝ 15.2Hz,2H),1.94(t,J＝19.0Hz,3H),1.73(dp,J＝26.2,6.6Hz,2H).
实施例47： 2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯 氧基)-6H-噻吩并[2,3-e]吲唑盐酸盐的制备
Figure PCTCN2022134097-appb-000146
制备方法类似于实施例43中的制备方法，不同的是将L-酒石酸替换为盐酸，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.55(s,1H),10.90(d,J＝66.9Hz,1H),8.39(s,1H),7.59–7.47(m,3H),7.39(d,J＝8.5Hz,1H),7.28–7.22(m,1H),6.87(d,J＝7.8Hz,2H),6.76(d,J＝10.7Hz,2H),4.95(d,J＝71.9Hz,1H),4.66–4.33(m,4H),4.10(d,J＝51.6Hz,2H),3.32–3.23(m,2H),2.04–1.84(m,5H).
实施例48： 2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-((1-(3-氟丙基)氮杂环丁烷-3-基)氧基)苯 氧基)-6H-噻吩并[2,3-e]吲唑磷酸盐的制备
Figure PCTCN2022134097-appb-000147
制备方法类似于实施例43中的制备方法，不同的是将L-酒石酸替换为磷酸，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ8.39(s,1H),7.56–7.52(m,2H),7.49(dd,J＝9.9,2.8Hz,1H),7.37(td,J＝8.3,2.8Hz,1H),7.25(d,J＝8.9Hz,1H),6.84–6.80(m,2H),6.72–6.67(m,2H),4.69(p,J＝5.6Hz,1H),4.45(dt,J＝47.4,6.0Hz,2H),3.78(t,J＝7.1Hz,2H),3.07–2.98(m,2H),2.63–2.54(m,2H),1.94(t,J＝19.0Hz,3H),1.68(dp,J＝25.8,6.5Hz,2H).
实施例49：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(6-氟-2-氮杂螺[3.3]庚烷-2-基)乙氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000148
制备方法类似于实施例7的制备方法，不同的是将步骤5中的原料3-(氟甲基)氮杂环丁烷盐酸盐替换为6-氟-2-氮杂螺[3.3]庚烷盐酸盐，制得标题化合物。 1H NMR(500MHz,DMSO-d 6)δ13.54(s,1H),8.39(s,1H),7.56–7.51(m,2H),7.48(dd,J＝9.9,2.8Hz,1H),7.35(td,J＝8.4,2.9Hz,1H),7.26(d,J＝8.9Hz,1H),6.84–6.78(m,2H),6.77–6.73(m,2H),4.90(dp,J＝56.0,6.7Hz,1H),3.78(t,J＝5.5Hz,2H),3.16(s,2H),3.14(s,2H),2.62(t,J＝5.5Hz,2H),2.48–2.41(m,2H),2.23–2.12(m,2H),1.94(t,J＝19.0Hz,3H).MS(ESI-LR)m/z:582.4(M+1) +.HRMS(ESI):Anal.Calcd for C 31H 28F 4N 3O 2S,582.1833；found 582.1842.
实施例50：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(1-(3-氟丙基)氮杂环丁烷-3-硫代)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：3-((4-((6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-氧基)苯基)硫代)氮杂环丁烷-1-羧酸叔丁酯
Figure PCTCN2022134097-appb-000149
将实施例13中步骤1所得3-(4-碘苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑(356mg，0.74mmol)，3-巯基氮杂环丁烷-1-羧酸叔丁酯(182mg，0.97mmol)，三(二亚苄基丙酮)二钯(17mg，0.019mmol)，4,5-双二苯基膦-9,9-二甲基氧杂蒽(21mg，0.037mmol)和N,N-二异丙基乙胺(0.193mL，1.11mmol)溶于1,4-二氧六环(5mL)，氩气保护，微波160℃反应1.5h，反应结束后向反应液中加入水(10mL)，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和氯化钠溶液(20mL)洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-3/1)纯化后，浓缩得白色泡沫299mg，收率为66.2％。 1H NMR(400MHz,CDCl 3)δ8.21(s,1H),7.61(d,J＝8.9Hz,1H),7.57(d,J＝8.9Hz,1H),7.32–7.27(m,2H),7.08–7.03(m,2H),6.67(s,1H),5.79(dd,J＝9.3,2.7Hz,1H),4.30–4.25(m,2H),4.07–4.00(m,1H),3.96–3.84(m,3H),3.80–3.72(m,1H),2.65–2.54(m,1H),2.22–2.08(m,2H),1.84–1.66(m,3H),1.43(s,9H).
步骤2：3-(2-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2，3-e]吲唑-3-基)氧基)苯基硫代)氮杂环丁烷-1-羧酸叔丁酯的制备
Figure PCTCN2022134097-appb-000150
将上步得到的3-((4-((6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-氧基)苯基)硫代)氮杂环丁烷-1-羧酸叔丁酯(280mg，0.52mmol)溶于乙腈(5mL)中，随后加入N-溴代丁二酰亚胺(102mg，0.57mmol)，氩气保护。避光条件下，室温搅拌过夜。饱和硫代硫酸钠溶液(10mL)淬灭反应，用乙酸乙酯(10mL×3)萃取，合并有机相，用饱和氯化钠溶液(10mL)洗涤，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-5/1)纯化后，浓缩得无色油状物272mg，收率84.8％。 1H NMR(400MHz,CD 3OD)δ8.28(s,1H),7.66(d,J＝9.0Hz,1H),7.41(d,J＝9.0Hz,1H),7.38–7.32(m,2H),6.96–6.91(m,2H),5.86(dd,J＝9.6,2.6Hz,1H),4.27(t,J＝8.5Hz,2H),4.03–3.95(m,2H),3.85–3.73(m,3H),2.58–2.45(m,1H),2.20–2.01(m,2H),1.92–1.69(m,3H),1.41(s,9H).
步骤3：3-((4-(2-(2-(1，1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2，3-e]吲唑-3-氧基)苯基)硫代)氮杂环丁烷-1-羧酸叔丁酯的制备
Figure PCTCN2022134097-appb-000151
将上步得到的3-(2-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-基)氧基)苯基硫代)氮杂环丁烷-1-羧酸叔丁酯(250mg，0.405mmol)，2-(2-(1,1-二氟乙基)-4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷(232mg，0.81mmol)，三(二亚苄基丙酮)二钯(37mg，0.041mmol)，2-二环己基膦-2',4',6'-三异丙基联苯(39mg，0.081mmol)，磷酸钾(258mg，1.22mmol)溶于1,4-二氧六环(5mL)和水(0.5mL)的混合溶剂中，110℃微波反应2h。反应结束后，直接将反应液旋干，硅胶拌样，经柱层析(石油醚/乙酸乙酯＝10/1-3/1)纯化后，浓缩得淡黄色泡沫169mg，收率为60％。 1H NMR(400MHz,CDCl 3)δ8.23(d,J＝0.8Hz,1H),7.56(dd,J＝9.0,0.9Hz,1H),7.40–7.31(m,3H),7.19–7.13(m,2H),7.06(td,J＝8.1,2.8Hz,1H),6.83–6.78(m,2H),5.80(dd,J＝9.4,2.6Hz,1H),4.23(dd,J＝8.6,7.1Hz,2H),4.04(dt,J＝10.7,3.2Hz,1H),3.90–3.71(m,4H),2.64–2.53(m,1H),2.24–2.08(m,2H),1.89(t,J＝18.5Hz,3H),1.83–1.64(m,3H),1.42(s,9H).
步骤4：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(1-(3-氟丙基)氮杂环丁烷-3-硫代)苯氧基)-6H-噻吩并[2,3-e]吲唑
Figure PCTCN2022134097-appb-000152
制备方法类似于实施例14中步骤5-6的制备方法，不同的是将步骤5中的2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(1-叔丁氧羰基氮杂环丁烷-3-氧基)苯氧基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑替换为3-((4-(2-(2-(1，1-二氟乙基)-4-氟苯基)-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-3-氧基)苯基)硫代)氮杂环丁烷-1-羧酸叔丁酯，制得标题化合物。 1H NMR(400MHz,DMSO-d 6)δ13.53(s,1H),8.40(s,1H),7.59–7.52(m,2H),7.49(dd,J＝9.9,2.8Hz,1H),7.37(td,J＝8.3,2.7Hz,1H),7.24(d,J＝8.9Hz,1H),7.19–7.13(m,2H),6.92–6.85(m,2H),4.40(dt,J＝47.4,6.0Hz,2H),3.84(p,J＝6.8Hz,1H),3.58(t,J＝7.3Hz,2H),2.88(t,J＝7.0Hz,2H),2.39(t,J＝7.0Hz,2H),1.95(t,J＝19.0Hz,3H),1.58(dp,J＝25.6,6.5Hz,2H).MS(ESI-LR)m/z:572.1(M+1) +.HRMS(ESI):Anal.Calcd for C 29H 26F 4N 3OS 2,572.1448；found 572.1457.
实施例51：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-(3-氟丙基)-2-氮杂螺[3.3]庚烷-6-基)氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000153
制备方法类似于实施例14的制备方法，不同的是将步骤3中的原料3-碘氮杂环丁烷-1- 羧酸叔丁酯替换为6-碘-2-氮杂螺[3.3]庚烷-2-羧酸叔丁酯，制得标题化合物。 1H NMR(500MHz,CD 3OD)δ8.25(s,1H),7.47(d,J＝8.9Hz,1H),7.39(d,J＝9.0Hz,1H),7.37–7.31(m,2H),7.11–7.04(m,1H),6.71(d,J＝8.5Hz,2H),6.60(d,J＝8.5Hz,2H),4.50–4.35(m,3H),3.39(s,2H),3.34(s,2H),2.67–2.56(m,4H),2.20–2.13(m,2H),1.86(t,J＝18.6Hz,3H),1.72(dt,J＝26.2,6.9Hz,2H).MS(ESI-LR)m/z:596.3(M+1) +.HRMS(ESI):Anal.Calcd for C 32H 30F 4N 3O 2S,596.1993；found 596.1989.
实施例52：2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(2-氟乙基)-2-氮杂螺[3.3]庚烷-6-氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000154
制备方法类似于实施例14的制备方法，不同的是将步骤3中的原料3-碘氮杂环丁烷-1-羧酸叔丁酯替换为6-碘-2-氮杂螺[3.3]庚烷-2-羧酸叔丁酯，将步骤6中的原料1-氟-3-碘丙烷替换为1-氟-2-碘乙烷，制得标题化合物。MS(ESI-LR)m/z:582.3(M+1) +.HRMS(ESI):Anal.Calcd for C 31H 28F 4N 3O 2S,582.1833；found 582.1837.
实施例53：2-(2-(1,1-二氟乙基)-4-氟苯基)-8-氟-3-(4-(1-(3-氟丙基)氮杂环丁烷-3-氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：3,4-二氟吲唑-5-甲腈的制备
Figure PCTCN2022134097-appb-000155
将4-氟-1H-吲唑-5-甲腈(5.16g，32mmol)溶于乙腈(100mL)中，加入1-氯甲基-4-氟-1,4-二氮杂双环[2.2.2]辛烷二(四氟硼酸)盐(22.688g，64mmol)，室温反应16小时后，补加1-氯甲基-4-氟-1,4-二氮杂双环[2.2.2]辛烷二(四氟硼酸)盐(11.344g，32mmol)，继续反应32小时后，补加1-氯甲基-4-氟-1,4-二氮杂双环[2.2.2]辛烷二(四氟硼酸)盐(5.672g，16mmol)，反应12小时。反应结束后，将反应液用硅藻土过滤，乙酸乙酯淋洗，滤液浓缩，硅胶拌样旋干。粗品经柱层析(石油醚/乙酸乙酯＝10/1-3/1)纯化后，浓缩得黄白色粉末0.9g，收率为15.7％。 1H NMR(400MHz,DMSO-d 6)δ13.47(s,1H),7.75(dd,J＝8.8,6.1Hz,1H),7.51(dd,J＝8.8,2.2Hz,1H).
步骤2：3,4-二氟-1-四氢吡喃-2-基-1H-吲唑-5-甲腈的制备
Figure PCTCN2022134097-appb-000156
将上步所得3,4-二氟吲唑-5-甲腈(100mg，0.558mmol)溶于二氯甲烷(5mL)，随后加入对甲苯磺酸一水合物(11mg，0.056mmol)和3,4-二氢-2H-吡喃(0.15mL，1.675mmol)，加热至40℃反应24h。反应结束后，加水(10mL)，用二氯甲烷(10mL×3)萃取，合并有机相，用饱和氯化钠溶液(10mL)洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝10/1-5/1)纯化后，浓缩得浅黄色油状物124mg，收率为 84.7％。 1H NMR(400MHz,CDCl 3)δ7.51(dd,J＝8.9,5.6Hz,1H),7.41(dd,J＝8.9,1.8Hz,1H),5.61(dt,J＝8.3,2.4Hz,1H),4.00–3.93(m,1H),3.77–3.67(m,1H),2.44–2.32(m,1H),2.18–2.01(m,2H),1.79–1.66(m,3H).
步骤3：3-氨基-8-氟-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑-2-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000157
将上步所得的3,4-二氟-1-四氢吡喃-2-基-1H-吲唑-5-甲腈(1.6g，6.08mmol)溶于N,N-二甲基甲酰胺(20mL)，加入碳酸钾(3.36g，24.32mmol)和巯基乙酸甲酯(0.6mL，6.68mmol)，升温至65℃，搅拌反应15h。反应结束后，将反应液倾倒至搅拌的冰水中(50mL)，析出大量固体，继续搅拌10分钟。随后将反应液抽滤，滤饼用水淋洗2-3遍，固体红外干燥，得浅橙色粉末1.72g，收率为81％。 1H NMR(400MHz,DMSO-d 6)δ8.19(d,J＝9.2Hz,1H),7.78(dd,J＝9.2,2.0Hz,1H),7.29(s,2H),5.88(d,J＝9.7Hz,1H),3.93–3.86(m,1H),3.82–3.70(m,4H),2.34–2.19(m,1H),2.06–1.92(m,2H),1.80–1.64(m,1H),1.62–1.51(m,2H).
步骤4：3-溴-8-氟-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑-2-羧酸甲酯的制备
Figure PCTCN2022134097-appb-000158
将上步所得的3-氨基-8-氟-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑-2-羧酸甲酯(1.7g，4.86mmol)，CuBr 2(1.62g，7.3mmol)溶于乙腈(20mL)中，氩气保护下升温至65℃，随后在加热条件下滴加亚硝酸叔丁酯(0.93mL，7.3mmol)，继续加热搅拌反应1h。反应结束后将反应液冷却至室温，倒至冰水中(50mL)，搅拌析出大量固体。随后将反应液抽滤，滤饼用水淋洗2-3遍，固体红外干燥。将固体悬浮于石油醚/乙酸乙酯＝5/1的混合溶剂(20mL)中，搅拌打浆0.5h，倾析除去上层清液，固体重复打浆步骤3次后，抽滤，滤饼用石油醚淋洗，红外干燥，得土黄色粉末1.16g，收率为57.8％。 1H NMR(400MHz,CDCl 3)δ7.95(d,J＝9.2Hz,1H),7.62(d,J＝9.2Hz,1H),5.69(d,J＝8.5Hz,1H),4.06–3.93(m,4H),3.83–3.68(m,1H),2.55–2.40(m,1H),2.22–2.02(m,2H),1.81–1.64(m,3H).
步骤5：3-溴-8-氟-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000159
将上步所得的3-溴-8-氟-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑-2-羧酸甲酯(1.1g，2.66mmol)，氧化亚铜(228mg，1.6mmol)和氢氧化锂一水合物(279mg，6.65mmol)溶于N,N-二甲基甲酰胺(15mL)，130℃加热搅拌反应3h。反应结束后，冷至室温，加入乙酸乙酯(20mL)稀释，硅藻土过滤，滤饼用乙酸乙酯淋洗(30mL)。随后向滤液中加水(30mL)，用乙酸乙酯(20mL×3)萃取，合并有机相，用水(30mL×3)和饱和氯化钠溶液(30mL)依次洗涤有机相，无水硫酸钠干燥，过滤，减压蒸馏除去溶剂。粗品经柱层析(石油醚/乙酸乙酯＝50/1-10/1)纯化后，浓缩得白色粉末693mg，收率为73.3％。 1H NMR(400MHz,CDCl 3)δ7.82(d,J＝9.1Hz,1H),7.57(dd,J＝9.1,2.1Hz,1H),7.41(s,1H),5.68(dt,J＝9.1,2.6Hz,1H),4.06–3.97 (m,1H),3.78–3.71(m,1H),2.56–2.43(m,1H),2.22–2.04(m,2H),1.83–1.62(m,3H).
步骤6：3-溴-8-氟-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑-1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000160
将上步所得的3-溴-8-氟-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑(5.3g，15.71mmol)为原料，按照中间体B1制备方法中的步骤3制备，得黄白色粉末647mg，收率为91.3％。 1H NMR(400MHz,DMSO-d 6)δ8.24(dd,J＝8.9,2.0Hz,1H),8.07(s,1H),7.77(d,J＝8.9Hz,1H),5.97(dt,J＝9.3,2.5Hz,1H),3.92–3.84(m,1H),3.81–3.71(m,1H),2.30–2.16(m,1H),2.06–1.94(m,2H),1.79–1.67(m,1H),1.63–1.49(m,2H).
步骤7:2-(2-(1,1-二氟乙基)-4-氟苯基)-8-氟-3-(4-(1-(3-氟丙基)氮杂环丁烷-3-氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
Figure PCTCN2022134097-appb-000161
制备方法类似于实施例14的制备方法，不同的是将步骤1的起始原料3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑1,1-二氧化物替换为上步所得的3-溴-8-氟-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑1,1-二氧化物，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.05(s,1H),7.54(dd,J＝8.6,5.7Hz,1H),7.51–7.46(m,2H),7.37(td,J＝8.4,2.7Hz,1H),7.33(d,J＝9.0Hz,1H),6.84–6.80(m,2H),6.71–6.66(m,2H),4.65(p,J＝5.7Hz,1H),4.44(dt,J＝47.4,6.0Hz,2H),3.67(t,J＝6.8Hz,2H),2.90(t,J＝6.7Hz,2H),2.53–2.49(m,2H),1.94(t,J＝19.0Hz,3H),1.65(dp,J＝25.7,6.5Hz,2H).MS(ESI-LR)m/z:574.1(M+1) +.
实施例54：8-氯-2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(1-(3-氟丙基)氮杂环丁烷-3-氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑的制备
步骤1：3-氯-4-氟-1H-吲唑-5-甲腈的制备
Figure PCTCN2022134097-appb-000162
将4-氟-1H-吲唑-5-甲腈(9.67g，60mmol)溶于乙醇(100mL)中，0℃下滴加11-14％的次氯酸钠溶液(88mL，300mmol)，室温反应5小时。反应结束后，加水(100mL)稀释，用1N稀盐酸调节pH至6左右。将反应液过滤，用水(50mL)淋洗，红外干燥得9.55g浅橙色粉末，收率为81.3％。 1H NMR(400MHz,DMSO-d 6)δ14.12(s,1H),7.76(dd,J＝8.8,5.9Hz,1H),7.57(d,J＝8.8Hz,1H).
步骤2：3-溴-8-氯-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑-1,1-二氧化物的制备
Figure PCTCN2022134097-appb-000163
其制备方法类似于实施例53中所述的步骤2至步骤6，不同的是将起始原料3,4-二氟吲唑-5-甲腈替换为3-氯-4-氟-1H-吲唑-5-甲腈，制得标题化合物。 1H NMR(400MHz, DMSO-d 6)δ8.28(d,J＝8.8Hz,1H),8.05(s,1H),7.80–7.73(m,1H),6.02(dd,J＝9.4,2.2Hz,1H),3.91–3.84(m,1H),3.80–3.74(m,1H),2.37–2.24(m,1H),2.07–1.97(m,2H),1.80–1.67(m,1H),1.64–1.54(m,2H).
步骤3：8-氯-2-(2-(1,1-二氟乙基)-4-氟苯基)-3-(4-(1-(3-氟丙基)氮杂环丁烷-3-氧基)苯氧基)-6H-噻吩并[2,3-e]吲唑
Figure PCTCN2022134097-appb-000164
制备方法类似于实施例14的制备方法，不同的是将步骤1的起始原料3-溴-6-(四氢-2H-吡喃-2-基)-6H-噻吩并[2,3-e]吲唑-1,1-二氧化物替换为上步所得的3-溴-8-氯-6-四氢吡喃-2-基-6H-噻吩并[2,3-e]吲唑-1,1-二氧化物，制得标题化合物。 1H NMR(600MHz,DMSO-d 6)δ13.71(s,1H),7.57–7.53(m,2H),7.49(dd,J＝9.9,2.8Hz,1H),7.37(td,J＝8.3,2.7Hz,1H),7.34(d,J＝8.9Hz,1H),6.85–6.80(m,2H),6.71–6.66(m,2H),4.65(p,J＝5.7Hz,1H),4.44(dt,J＝47.4,6.1Hz,2H),3.66(td,J＝6.1,1.9Hz,2H),2.88(dd,J＝8.0,5.5Hz,2H),2.47(t,J＝7.4Hz,2H),1.94(t,J＝19.0Hz,3H),1.64(dp,J＝25.7,6.5Hz,2H).MS(ESI-LR)m/z:590.0(M+1) +.
本发明所述的代表性化合物的药理学活性评价
实验一：本发明代表性化合物体外抗乳腺癌MCF-7细胞增殖活性评价
通过CTG法检测化合物对MCF-7细胞增殖的影响并计算化合物的IC 50，CellTiter-Glo简称CTG，是通过对ATP进行定量来测定培养物中活细胞数目的一种快速均质检测方法。ATP是活细胞新陈代谢的关键指标，均质的检测使得细胞裂解和产生的发光信号与存在的ATP量成正比，而ATP量直接与培养物中的细胞数量成正比。
MCF-7细胞用DMEM+10％FBS培养。细胞置于37℃孵箱中5％CO 2条件下培养。细胞传代、复苏及冻存均按常规方法进行。MCF-7细胞生长至80％左右聚合度，胰蛋白酶消化细胞，以每孔4×10 3个细胞接种到96孔板中，接种体积为每孔90μL，37℃孵箱中培养过夜。第二天，加入浓度梯度的药物，每孔10μL。化合物最高检测浓度为10μM，4倍稀释，10个浓度。将96孔板置于5％CO 2，37℃孵箱中培养6天。96孔培养板中每孔加入50μL的CTG试剂，在振荡器上震荡5min，室温避光放置10min，取60μL转移至384孔Opti-plate。使用多功能酶标仪读取发光(luminescence)信号。信号强度用于表征活细胞数量。各加药组数据以对照组均值为100％进行标化，用GraphPad Prism软件按以下公式进行曲线拟合及IC 50(半数抑制浓度)计算：
Y＝Bottom+(Top-Bottom)/(1+10^((LogIC 50-X)*HillSlope))
表1：本发明代表性化合物体外抗乳腺癌MCF-7细胞增殖活性
化合物 IC 50(nM) 化合物 IC 50(nM) 化合物 IC 50(nM)
LSZ102 4.22 SAR439859 9.07 实施例1 1.92
实施例2 1.54 实施例7 1.06 实施例8 2.38
实施例9 3.25 实施例14 1.01 实施例15 2.56
实施例16 0.93 实施例19 2.23 实施例20 1.13
实施例21 0.75 实施例23 0.58 实施例24 0.61
实施例25 0.82 实施例26 1.12 实施例27 1.02
实施例28 1.74 实施例31 3.09 实施例32 1.67
实施例33 1.7 实施例34 0.66 实施例35 1.15
实施例36 1.44 实施例37 0.77 实施例38 2.83
实施例39 1.44 实施例40 1.57 实施例41 0.55
实施例42 0.48 实施例49 2.26 实施例52 1.18
实施例53 0.89        
从表1的实验结果可见，本发明的化合物对MCF-7乳腺癌细胞增殖有很强的抑制活性，且抗增殖活性明显优于临床I期对照药物LSZ102，远远优于临床III期对照药物SAR439859。特别是实施例42，41，23，24化合物抗肿瘤活性比对照LSZ-102和SAR439859分别提高7倍和15倍以上。
实验二：本发明代表性化合物体外抗他莫昔芬耐药乳腺癌细胞MCF-7 Tam1增殖活性评价
MCF-7 Tam1细胞通过MCF-7细胞长期暴露于他莫昔芬的活性代谢产物4-羟基他莫昔芬，同时伴有长期雌激素剥夺的培养条件而建立的，具有对他莫昔芬和芳香化酶抑制剂的抗性。
MCF-7 Tam1细胞传代培养，培养条件为含有青霉素(终浓度为100U/mL)、链霉素(终浓度为100μg/mL)、10μg/mL人胰岛素、1μM 4-羟基他莫昔芬以及10％FBS的DMEM培养基，当细胞融合至90％时，弃去旧培养基，用2ml PBS洗涤细胞2次，弃去PBS后加入2mL 0.25％胰蛋白酶-0.02％EDTA混合消化液，置显微镜下观察，约30s，当细胞变圆后迅速加入2ml完全培养基终止消化，轻轻吹打，收集细胞。800rpm，4℃，离心5min，弃去上清，用完全培养基重悬细胞，分瓶培养，隔天换液。化合物最高检测浓度为10μM，10倍稀释，8个浓度。使用适合进行化学发光检测的96孔板，每孔接种100μl细胞(1×10 4cells/ml)。按试验分组将各组细胞使用含对应浓度化合物的培养基孵育5天。取出细胞培养板，室温平衡10min。96孔板每孔加入100μl CTG检测试剂，室温孵育10min。酶标仪检测各组RFU值。抑制率计算方式为：抑制率＝(对照组OD值-给药组OD值)/对照组OD值。使用GraphPad计算各化合物对MCF-7 Tam1细胞的IC 50。
表2：本发明代表性化合物体外抗他莫昔芬耐药乳腺癌细胞MCF-7 Tam1增殖活性
化合物 IC 50(nM)
LSZ102 9.6
SAR439859 17.63
实施例14 3.19
实施例20 5.2
实施例50 2.53
如图1和表2所示，与LSZ102和SAR439859相比，本发明实施例14、实施例20和实施例50均对MCF-7 Tam1有更强的增殖抑制作用。特别是实施例14和实施例50，其抗 肿瘤活性是LSZ102的3倍以上，是SAR439859的5倍以上，具备强效抗MCF-7耐他莫昔芬乳腺癌细胞增殖的能力。
实验三：本发明代表性化合物体外基于细胞水平的ERα降解活性评价
通过基于细胞的高内涵成像方法分析本发明中所述的化合物引起的ER降解的程度。
MCF-7细胞培养基：88％RPMI 1640,10％FBS,1％P/S和1％GlutaMax.
MCF-7播种培养基：88％RPMI 1640,10％FBS,1％P/S和1％GlutaMax.
第一天：用细胞播种培养基稀释细胞悬浮液至8.75×10 4cells/mL，随后将40μL细胞悬液滴入每孔检测板，置于37℃CO 2培养箱中。第三天：化合物稀释4倍连续稀释，转移500μL，10个浓度点。用20μL培养基稀释化合物，用Bravo转10μL至板上，在37℃培养箱中培养24小时。第四天：在测定板中加入50μL 8％多聚甲醛，室温下放置40分钟。每孔用100μL PBS洗涤检测板两次，然后在检测板中加入50μL 0.1％TritonX-100的PBS，室温下放置15分钟。每孔用100μL PBS洗涤检测板5次，然后加入含0.1％Tween 20的封闭缓冲液50μL，室温下放置1小时。弃掉检测板中的溶液，用阻断缓冲液按1:1000的比例稀释一抗，加入25μL检测板，4℃孵育过夜。第五天：每孔用100μL PBS洗涤检测板5次，然后用阻断缓冲液稀释二抗(1:1000)和DRAQ5(1:2000)，加入25μL至检测板，室温孵育1小时。每孔用100μL PBS冲洗检测板5次，在Odyssey红外成像系统上读取检测板。然后将数据标准化，将800通道(ER)的综合强度除以700通道(DNA)的综合强度。背景减法是将所有归一化实验值减去归一化阴性对照井(无一抗体)的平均值。然后通过将每个实验值除以DMSO控制值的平均值(％响应＝(实验值/DMSO控制值)×100)计算归一化/背景减除数据的百分比响应。使用GraphPad Prism v6.02软件(GraphPad,San Diego,CA)生成剂量响应曲线、EC 50和百分比ERα剩余值。
表3：本发明代表性化合物体外ERα降解活性
化合物 EC 50(nM) 化合物 EC 50(nM) 化合物 EC 50(nM)
LSZ102 1.01 SAR439859 1.7 实施例1 1.18
实施例7 1.25 实施例8 0.41 实施例9 0.83
实施例14 1.13 实施例15 0.69 实施例16 0.40
实施例20 1.16 实施例21 1.28 实施例23 1.07
实施例24 0.95 实施例27 1.35 实施例34 1.27
实施例37 1.29 实施例41 1.78 实施例42 1.41
从表3的实验结果可见，本发明的化合物对MCF-7细胞内ERα蛋白具有明显的降解作用，且降解活性和临床I期药物LSZ102相当或更优，远优于临床III期对照药物SAR439859，其中实施例8，9，15，16化合物相比SAR439859提高了2-4倍活性。
实验四：MCF-7细胞荧光报告基因实验检测化合物对野生型ERα拮抗活性
ERα报告基因构建原理：HEK293/GAL4/ERα，细胞株利用分子克隆方法将雌激素受体LBD区域(化合物结合区域)和GAL4DBD区域(DNA结合区域)进行融合蛋白表达。当配体激活ERα时，ERα-GAL4融合蛋白启动下游luciferase基因表达，读板机检测到化学发光信号。ERα配体刺激浓度和化学发光信号呈剂量依赖关系。
将HEK293/GAL4/ERα细胞悬液收集起来，1000转/分钟离心5分钟，去掉上清，用 预热的培养基(DMEM(无酚红)，含碳吸附血清10％，即500mL细胞培养基中包含450mL DMEM，50mL碳吸附血清)重悬，计数后用培养基稀释细胞悬液，按照40000细胞/孔接种到96孔细胞培养板中，每孔接种80μL细胞悬液，37℃，5％CO 2培养箱，孵育过夜。实验当天，每孔加入10μL化合物工作液到细胞板中，37℃，5％CO 2培养箱，继续孵育1小时，再每孔加入10μL含有激动剂的培养基(10nM Estradiol)，Estradiol终浓度为1nM，DMSO终浓度为0.5％。细胞板在37℃，5％CO 2培养箱中继续孵育24小时。结束孵育后，去掉细胞上清，细胞板每孔加入50μL Bright Glo检测试剂，25℃，孵育2分钟。结束孵育后，使用EnVision进行发光信号检测。
数据根据如下公式计算化合物处理后的抑制率：％抑制率＝100-(RFU化合物–RFU空白对照)/(RFU阴性对照–RFU空白对照)×100％。阴性对照：激动剂处理的细胞；空白对照：无激动剂处理的细胞，然后利用Prism作图计算化合物的IC 50值。
表4：本发明代表性化合物对 野生型ERα拮抗活性
化合物 IC 50(nM) 化合物 IC 50(nM) 化合物 IC 50(nM)
LSZ102 6.53 SAR439859 1.8 a 实施例1 1.55
实施例7 0.72 实施例14 0.91 实施例16 0.61
实施例20 1.05 实施例21 1.07 实施例23 0.60
实施例24 0.61 实施例25 1.3 实施例27 0.97
实施例37 0.97 实施例41 0.89 实施例42 0.75
a数据来源于Mol.Cancer Ther.2021,20,250-262.
从表4的实验结果可见，本发明的化合物对野生型ERα有很强的拮抗作用，明显优于临床I期药物LSZ102和临床III期药物SAR439859。其中实施例16，23，24化合物对野生型ERα的拮抗活性相比LSZ102提高10倍以上，实施例7，14，20，21，27，37，41，42化合物对野生型ERα的拮抗活性相比LSZ-102提高6倍以上。
综合实验一、二、三、四结果表明本发明所述化合物对ERα具有很强的拮抗/降解双重功能，并且对雌激素依赖的MCF-7细胞和他莫昔芬耐药的MCF-7 Tam1细胞增殖均具有显著的抗增殖活性，表现出了极强的抗肿瘤药效，且明显优于临床对照化合物LSZ102和SAR439859。
实验五：本发明代表性化合物体外对ERα突变体Y537S和D538G的拮抗作用
与野生型ERα不同，ERα配体结合域中的Y537S和D538G突变导致在没有雌二醇的情况下自发募集共活化因子，例如过氧化物酶体增殖物激活受体-γ共激活因子和类固醇受体共激活因子，证实这些突变导致ERα的组成型激活。在这种激活的构象中，突变的ERα对雌二醇的亲和力增加，而对拮抗剂的亲和力降低。
SK-BR-3细胞培养基：89％1640不含酚红，10％活性炭处理FBS和1％GlutaMax
第一天：1.将80μL细胞悬液和30000个细胞接种到检测板的每孔中，37℃5％CO 2孵育24小时。第二天：制备转染试剂，室温放置15分钟。测定板每孔加入10μL转染试剂，37℃5％CO 2孵育24h。第三天：将10μL培养基(100nMβ-雌二醇，10μL培养基)加入检测板，37℃5％CO 2孵育24h。第四天：1.从每孔中取出50μL培养基，将50μL 荧光素酶检测试剂加入检测板中，室温振荡20分钟，然后在Envision上读数。2.将50μL Stop & Glo试剂加入检测板，室温振荡20分钟，然后在Envision上读数。使用XL-fit软件(供应商：ID Business Solutions Ltd.，软件版本：XL fit 5.0)分析数据，％Effect＝(样品值–空白对照)/(阳性对照-空白对照)*100。
表5：本发明代表性化合物 体外对ERα突变体Y537S和D538G的拮抗活性
ER突变体 SAR439859 实施例14 实施例20
Y537S(nM) 57.67 10.25 13.13
D538G(nM) 17.22 16.08 16.55
从图2和表5的实验结果可见，实施例14和实施例20对ERα突变体Y537S和D538G有很强的拮抗作用，其中针对Y537S突变体实施例14和实施例20相比SAR43959具有4-5倍的活性提高，而针对D538G突变体实施例14和实施例20与SAR439859具有相当的拮抗活性。
实验六：本发明代表性化合物体外对ERα突变体(Y537S)降解活性评价
1.蛋白样品的制备
MCF-7(Y537S)细胞按照80万细胞/孔铺到6孔板中，每孔1mL。铺板第二天加入1mL化合物，给药终浓度为1000nM、100nM、10nM、1nM和0.1nM。药物处理4小时，收集细胞，提取总蛋白，进行BCA测定。将提取的蛋白上清与5ⅹ蛋白上样缓冲液充分混合，金属浴：100℃，10min，变性完后冷却至室温再放入-80℃保存。
2.电泳
将制备好的胶固定到电泳槽上，储液池中倒入电泳液。用微量加样器将制备好的蛋白样品和Marker加入上样孔，各样品总蛋白量为50μg。转膜条件：300mA 30min(ERα66kDa和a-Tublin 55kDa)。用含5％脱脂奶粉的TBST(封闭液)浸泡PVDF膜，37℃摇床封闭1.5h。用封闭液稀释相应的一抗，使PVDF膜浸泡于一抗孵育液中，4℃孵育过夜。TBST充分洗涤PVDF膜3次，5min/次。用TBST稀释HRP标记羊抗兔二抗按照1:4000稀释，使PVDF膜浸泡于二抗孵育液中，室温摇床孵育1h。TBST充分洗涤PVDF膜3次，5min/次。将ECL试剂中增强液与稳定的过氧化物酶溶液按1:1比例混匀，滴加工作液于PVDF膜上，显影。
实验结果如图3所示，与LSZ102相比，实施例14能够在更低浓度下(10nM-100nM vs100nM-1000nM)诱导更强的ERα(Y537S)突变蛋白的降解，表明本发明化合物具有治疗内分泌耐药的雌激素受体突变(Y537S)乳腺癌的潜力。
实验七：化合物药代动力学性质评价
小鼠/大鼠按体重随机分组，给药前1天禁食不禁水12～14h，给药后4h给食。每只动物每次通过眼眶取0.1mL血液，EDTAK2抗凝，采集时间点为：给予受试物后0,5,15,30min,1,2,4,6,8,24h。血液样本采集后置于冰上，并于30分钟之内离心分离血浆(离心条件：5000转/分钟，10分钟，4℃)。分析前存放于–80℃。数据采集及控制系统软件为Analyst1.5.1软件(Applied Biosystem)。图谱样品峰积分方式为自动积分；采用样品峰面积和内标峰面积的比值作为指标，和样品的浓度进行回归。回归方式：线性回归，权重系数为1/X 2。药代动力学参数用WinNonlin Professional v6.3(Pharsight,USA)用非房室模型分析处理。Cmax为 实测的最大血药浓度，血药浓度-时间曲线下面积AUC(0→t)由梯形法计算得到，Tmax为给药后血药浓度达峰时间。实验数据用“均数±标准差”(Mean±SD,n≥3)或“均数”(Mean,n＝2)表示。
表6：本发明部分代表性化合物药代动力学参数
Figure PCTCN2022134097-appb-000165
a数据来源于J.Med.Chem.2018,61,2837-2864.Dose:1mg/kg iv,3mg/kg p.o.
由表6可知，本发明化合物代谢性质优越，表现出良好的药物暴露和较低的清除率(远低于LSZ102文献报道的28mL/min/kg)，生物利用度达到66.6％，是对照药物LSZ102的5倍以上，预示本发明化合物能够口服给药实现抗肿瘤的目标。
在雌性SD大鼠组织分布实验中，将受试化合物溶解在0.5％MC中，并以20mg/kg(n＝3)的剂量口服给药。给药后2小时后取组织：血浆、全血、脑、乳腺、卵巢、子宫、肝、肾，动物安乐死放血后取组织，清洗干净后用50％甲醇按照1:4比例匀浆，匀浆液存于-80℃待分析。
表7：本发明代表性化合物(实施例47)在雌性大鼠中的组织分布
组织 血浆 全血 脑 卵巢 子宫 肝 肾 乳腺
组织药物浓度(ng/mL或ng/g) 609 480 442 4419 2166 21223 7103 2410
组织/血浆比 - 0.8 0.7 7.3 3.6 34.9 11.7 4.0
表7结果表明，口服20mg/kg后，与血浆药物浓度相比，乳腺组织中药物浓度高出了4倍，表达ER的其他性器官如子宫和卵巢分别达到3.6倍和7.3倍的暴露量，具有一定的靶组织分布倾向。更重要的是，本发明化合物能够穿透血脑屏障，在脑组织中发现具有与血浆中相近的药物浓度(B/P＝0.7)，表明本发明化合物具有治疗ER+脑转移乳腺癌的潜力。
实验八：化合物对小鼠子宫增重的影响评价
试验动物：SPF级雌性不成熟SD大鼠，21天龄，由常州卡文斯实验动物有限公司提供，饲养于SPF级饲养环境中，室内温度控制在23±2℃，自由饮食和摄水。动物总数30只。实验前适应性饲养3天。
实验分组：空白组：每天口服2％吐温-80/0.5％羟甲基纤维素(10mg/kg)，连续3天。空白+17α-乙炔雌二醇组：每天口服17α-乙炔雌二醇(0.1mg/kg)，连续3天。4-羟基他莫昔芬组：每天口服4-羟基他莫昔芬组(60mg/kg)，连续3天。实施例14组：每天口服实施例14(10mg/kg)，连续3天。实施例16组：每天口服实施例16(10mg/kg)，连续3天。实施例20组：每天口服实施例20(10mg/kg)，连续3天。
最后一次给药24h后，二氧化碳法处死大鼠，解剖取子宫，仔细剔除无关组织，用D-Hanks液洗涤2～3次，洗去血液，沥干水分保存，称取重量。
HE实验：1)将固定好的子宫组织切成4μm厚的切片，将切片放在烘箱中1h；2)将干燥的石蜡切片进行常规二甲苯脱蜡，下行梯度乙醇水化，蒸馏水洗；3)加入苏木精染色10min～30min，然后用流水洗去苏木精染液；4)1％盐酸乙醇褪色，见切片变红，颜色较浅即可，在放入自来水流水中使其恢复蓝色。5)伊红染0色1min，流水冲洗；6)切片经梯度酒精脱水干燥，二甲苯透明，中性树胶封片。7)随机选择一个视野使用显微镜(200×)拍照。数据组间统计学差异采用one-way ANOVA和Tukey’s检验，P值小于0.05认为有显著性差异。
表8：本发明代表性化合物对未成熟大鼠子宫重量的影响
组别 空白组 空白+17α-乙炔雌二醇 4-羟基他莫昔芬 实施例14 实施例20
平均体重(g) 53.19 54.3 54.59 50.92 57.89
平均子宫湿重(mg) 30.64 75.50 59.10 16.94 21.37
子宫湿重：体重 0.06％ 0.14％ 0.11％ 0.03％ 0.04％
实验结果如图4和表8所示，与空白对照相比，17α-乙炔雌二醇和4-羟基他莫昔芬(他莫昔芬体内的活性代谢形式)与体重的比值显著分别增加了约2.5倍和2倍，表明对子宫组织的ERα有激动作用。但是本发明实施例均展示出了有效的减小子宫重量的作用，其中实施例14使子宫湿重与体重的比值降低约2倍，表明对ERα有拮抗/反向激动作用。此外，用苏木精和伊红对子宫内膜进行染色，以进行组织学评估。17α-乙炔雌二醇和4-羟基他莫昔芬增加了子宫湿重，上皮细胞呈高柱状表型。这与空白对照和实施例14中上皮的低立方表型形成鲜明对比，证实了化合物是一个完全的拮抗剂，没有发生宫内膜癌的风险。
实验九：化合物对MCF-7小鼠皮下肿瘤模型的生长抑制评价
试验试剂：
胎牛血清(SH30070.03)(FBS,Hyclone,Logan,UT,USA)；青霉素(I9532)(Sigma,St.Louis,MO,USA)；链霉素(85886)(Sigma,St.Louis,MO,USA)；重组人胰岛素(91077C)(Sigma,St.Louis,MO,USA)；EMEM培养基(30-2003)(ATCC,Rockville,MD,USA)；胰蛋白酶(15090046)(Gibco,Grand Island,NY,USA)；HBSS(H6648)、DMSO(D8418)、PEG400(8074851000)、PEG300(8074841000)、PBS(806552)、Solutol HS-15(42966)(Sigma,St.Louis,MO,USA)；Matrigel Matrix(BD Bioscience,USA)；雌激素微丸(0.36mg雌二醇，60天释放)(SE-121)(Innovative Research of America,Florida,USA)；PVDF膜(0.45μm)(Millipore,Schwalbach,德国)；StarSignal Western Protein Marker(10-200kDa)(M227-01)(GenStar,北京,中国)；丽春红、吐温20、丙烯酰胺、十二烷基硫酸钠、PMSF(Solon,OH,USA)；蛋白印迹膜再生液(ZN1923,Biolab,北京,中国)；蛋白裂解液(RIPA)、1.5mol/L Tris HCl(pH6.8)、1.5mol/L Tris HCl(pH8.8)(Beyotime,上海,中国)；ECL发光液(Thermo Fisher Scientific,Pittsburgh,PA,USA)；Anti-ERα抗体(21244-1-AP)购自Proteintech公司(Proteintech,湖北,中国)；Goat Anti-Rabbit IgG H&L(HRP)(ab6721)购自Abcam公司(Abcam,Cambridge,UK)。
细胞的培养条件：
MCF-7细胞传代培养，培养条件为含有青霉素(终浓度为100U/mL)、链霉素(终浓度为100μg/mL)、human recombinant insulin(终浓度为0.01mg/mL)以及10％FBS的EMEM 培养基，当细胞融合至90％时，弃去旧培养基，用2ml PBS洗涤细胞2次，弃去PBS后加入2mL 0.25％胰蛋白酶-0.02％EDTA混合消化液，置显微镜下观察，约30s，当细胞变圆后迅速加入2ml完全培养基终止消化，轻轻吹打，收集细胞。800rpm，4℃，离心5min，弃去上清，用完全培养基重悬细胞，分瓶培养，隔天换液。
小鼠移植瘤模型构建：
在肿瘤植入前3天，用无菌的套管针将雌激素微丸(0.36mg雌二醇，60天释放)植入小鼠肩胛骨之间的皮下。取对数生长期的MCF-7细胞，使用胰蛋白酶将MCF-7细胞消化，使用50％HBSS和50％Matrigel混合液将MCF-7细胞重悬成10 7cells/mL细胞悬液。每只小鼠于右侧腋窝乳腺脂肪垫区域皮下注射200μL MCF-7细胞悬液，每3天测量肿瘤体积(宽 2×长×π/6)和体重；肿瘤平均体积达到约200mm 3时，将小鼠随机分组并灌胃给药。
试验分组：
空白组：10％PEG300+25％of 20％Solutol+65％PBS，小鼠MCF-7移植瘤模型建立且肿瘤平均体积达到约200mm 3后，每日灌胃给药，灌胃体积为0.1ml/10g，连续给药21天(N＝8)；
LSZ102(15mg/kg)组：小鼠MCF-7移植瘤模型建立且肿瘤平均体积达到约200mm 3后，每日灌胃给药LSZ102，给药剂量为15mg/kg，灌胃体积为0.1ml/10g，连续给药21天(N＝8)；
实施例14(15mg/kg)组：小鼠MCF-7移植瘤模型建立且肿瘤平均体积达到约200mm 3后，每日灌胃给药实施例14，给药剂量为15mg/kg，灌胃体积为0.1ml/10g，连续给药21天(N＝8)；
实施例14(5mg/kg)组：小鼠MCF-7移植瘤模型建立且肿瘤平均体积达到约200mm 3后，每日灌胃给药实施例14，给药剂量为5mg/kg，灌胃体积为0.1ml/10g，连续给药21天(N＝8)；
造模开始每3天早上固定时间称量各组小鼠体重并测量小鼠肿瘤体积大小(宽 2×长×π/6)；化合物的抑瘤疗效用肿瘤生长抑制率TGI(％)来评价。TGI(％)＝[(1-(某处理组给药结束时平均瘤体积-该处理组开始给药时平均瘤体积)/(溶剂对照组给药结束时平均瘤体积-溶剂对照组开始给药时平均瘤体积)]×100％。
表9：本发明代表性化合物皮下移植瘤模型肿瘤体积
Figure PCTCN2022134097-appb-000166
实验结果如图5和表9所示，和空白组相比，LSZ102(15mg/kg)、实施例14(15mg/kg)、实施例14(5mg/kg)组的小鼠MCF-7肿瘤体积显著下降(p<0.01)，实施例14(15mg/kg)组的肿瘤抑制率最高，对肿瘤的抑制作用最好，体内抗肿瘤活性约为LSZ102的3倍。
以上结果显示本发明的化合物可以通过拮抗/降解雌激素受体来治疗或预防与雌激素有关的多种疾病，例如癌症(乳腺癌、卵巢癌、结肠癌、前列腺癌、子宫内膜癌)，骨质疏松症，神经退行性疾病，心血管疾病，红斑狼疮，子宫内膜异位症及肥胖症等。
尽管以上已经对本发明作了详细描述，但是本领域技术人员理解，在不偏离本发明的精神和范围内的前提下可以对本发明进行各种修改和改变。本发明的权利范围并不限于上文所作的详细描述，而是如在所附权利要求中阐述的那样。

Claims (12)

  1. 一种通式(I)所示的化合物，或其前药、对映异构体、非对映异构体、外消旋体，或其药学上可接受的盐，
    Figure PCTCN2022134097-appb-100001
    式中，R 1选自：-COOH、-B(OH) 2、C1-C6烷基、C1-C6卤代烷基；
    R 2选自：H、卤素、氰基、C1-C6烷基、C1-C6卤代烷基；
    或者R 1、R 2与相连的苯环形成
    Figure PCTCN2022134097-appb-100002
    p为0、1、2、3或4；
    q为0、1、2或3；
    各R 3、各R 4独立地选自：氢、卤素、氰基、C1-C4烷基、C1-C4烷氧基、C1-C4卤代烷基、C1-C4卤代烷氧基、-SO 2NH 2、-C(O)NH 2；
    R 5、R 6各自独立地选自：氢、卤素、C1-C6烷基、C1-C6烷氧基、C1-C6卤代烷基、C1-C6卤代烷氧基，或R 5、R 6与相连的碳形成C3-C6碳环或3-6元杂环；
    R 7选自：卤素、C1-C6烷氧基、C1-C6卤代烷基、C1-C6卤代烷氧基；
    Z 1为-O-、-NH-、-S-、-CH 2O-、-CO-、-SO 2-、-SO-、-CH＝CH-、-C≡C-、C1-C4亚烷基；
    o为0、1、2、3或4；
    环Y为
    Figure PCTCN2022134097-appb-100003
    Z 2、Z 3各自独立地为-CH-或-N-；环A或环B相同或不同，各自独立地为C3-C6环烷基或3-6元杂环烷基；
    n为0、1、2、3或4；
    m为0、1、2、3或4；
    R 8为卤素、氰基、羟基、羧基或氨基；
    R 9选自卤素、氰基、氨基、C1-C6烷基、C1-C6卤代烷基。
  2. 如权利要求1所述的化合物，其特征在于，所述化合物具有下式：
    Figure PCTCN2022134097-appb-100004
    R 1、R 2、p、q、R 3、R 4、R 5、R 6、R 7、Z 1、o、Z 2、Z 3、n、m、R 8的定义如权利要求1所述。
  3. 如权利要求1或2所述的化合物，其特征在于，R 1选自：-COOH、-B(OH) 2、C1-C4烷基、C1-C4卤代烷基；
    R 2选自：H、卤素、氰基、C1-C4烷基、C1-C4卤代烷基；
    或者R 1、R 2与相连的苯环形成
    Figure PCTCN2022134097-appb-100005
    R 9选自卤素、氰基、氨基、C1-C2烷基、C1-C2卤代烷基。
  4. 如权利要求1或2所述的化合物，其特征在于，R 5、R 6各自独立地选自：氢、卤素、C1-C4烷基、C1-C4烷氧基、C1-C4卤代烷基、C1-C4卤代烷氧基，或R 5、R 6与相连的碳形成C3-C5碳环或3-5元杂环；
    R 7选自：卤素、C1-C4烷氧基、C1-C4卤代烷基、C1-C4卤代烷氧基。
  5. 如权利要求1或2所述的化合物，其特征在于，Z 1为-O-、-NH-、-S-、-CH 2O-、-CO-、-SO 2-、-SO-、-CH＝CH-、-C≡C-、C1-C2亚烷基。
  6. 如权利要求1所述的化合物，其特征在于，R 1为-COOH、-B(OH) 2；R 2为H；或者R 1、R 2与相连的苯环形成
    Figure PCTCN2022134097-appb-100006
    R 9选自氢、氟、氯、氰基、氨基、甲基、三氟甲基；
    p为0、1、2或3；各R 3独立地选自：氢、氟、氯、溴、氰基、C1-C2烷基、C1-C2烷氧基、C1-C2卤代烷基、C1-C2卤代烷氧基；
    q为0、1或2；各R 4独立地选自：氢、氟、氯、溴；
    R 5、R 6各自独立地选自：氢、氟、氯、溴、C1-C3烷基、C1-C2卤代烷基，或R 5、R 6与相连的碳形成C3-C5碳环；
    R 7选自：氢、氟、氯、溴、C1-C4烷基、C1-C2卤代烷基；
    Z 1为-O-、-NH-、-S-、-CH 2O-、-CO-、-SO 2-、-SO-、-CH＝CH-、-C≡C-、-CH 2-、-CH 2CH 2-；
    o为0、1、2或3；
    环Y为
    Figure PCTCN2022134097-appb-100007
    Z 2、Z 3各自独立地为-CH-或-N-；环A或环B相同或不同，彼此独立地代表C4环烷基或4元杂环烷基；
    n为1、2或3；
    m为0、1、2或3；
    R 8为氟、氯、溴或氰基。
  7. 如权利要求2所述的化合物，其特征在于，R 1为-COOH、-B(OH) 2；R 2为H；或者R 1、R 2与相连的苯环形成
    Figure PCTCN2022134097-appb-100008
    p为0、1、2或3；各R 3独立地选自：氢、氟、氯、溴、氰基、C1-C2烷基、C1-C2烷氧基、C1-C2卤代烷基、C1-C2卤代烷氧基；
    q为0、1或2；各R 4独立地选自：氢、氟、氯、溴；
    R 5、R 6各自独立地选自：氢、氟、氯、溴、C1-C3烷基、C1-C2卤代烷基，或R 5、R 6与相连的碳形成C3-C5碳环；
    R 7选自：氢、氟、氯、溴、C1-C4烷基、C1-C2卤代烷基；
    Z 1为-O-、-NH-、-S-、-CH 2O-、-CO-、-SO 2-、-SO-、-CH＝CH-、-C≡C-、-CH 2-、-CH 2CH 2-；
    o为0、1、2或3；
    Z 2、Z 3各自独立地为-CH-或-N-；
    n为1、2或3；
    m为0、1、2或3；
    R 8为氟、氯、溴或氰基。
  8. 如权利要求1所述的化合物，其特征在于，所述化合物或其药学上可接受的盐选自下组：
    Figure PCTCN2022134097-appb-100009
    Figure PCTCN2022134097-appb-100010
    Figure PCTCN2022134097-appb-100011
  9. 如权利要求1所述的化合物的制备方法，其特征在于，R 1为-B(OH) 2，R 2为氢，所述制备方法包括以下步骤：
    Figure PCTCN2022134097-appb-100012
    (i1)A1与有机硼试剂发生选择性Miyaura偶联反应得到A2，其中，所述的有机硼试剂选自：硼酸、联硼酸频那醇酯、频那醇硼烷、联硼酸新戊二醇酯；
    (i2)A2氧化开环得到A3；
    (i3)A3与A4发生亲核取代反应得到A5所示的式I化合物，
    各式中R 3、R 4、R 5、R 6、R 7、R 8、Z 1、Z 3、m、n、o、p、q如权利要求1所述；
    W，X分别为离去基团，各自独立地选自：卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基，
    或者R 1为-COOH，R 2为氢，所述制备方法包括以下步骤：
    Figure PCTCN2022134097-appb-100013
    (ii1)B1与B2发生亲核加成消除反应得到B3；
    (ii2)B3经过还原溴化得到B4；
    (ii3)B4与B5发生Suzuki偶联反应得到B6；
    (ii4)B6脱除保护基得到B7；
    (ii5)B7与B8发生亲核取代反应得到B9；
    (ii6)B9经Pinnick氧化得到B10所示的式I化合物；
    各式中R 3、R 4、R 5、R 6、R 7、R 8、Z 1、Z 3、m、n、o、p、q如权利要求1所述；
    W为离去基团，选自：卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基；
    或者R 1、R 2与相连的苯环形成
    Figure PCTCN2022134097-appb-100014
    时，所述制备方法包括以下步骤：
    Figure PCTCN2022134097-appb-100015
    (iii1)中间体int1与C1发生亲核加成消除反应得到C2；
    (iii2)C2经过砜基还原得到C3；
    (iii3)C3通过催化氢化反应脱除苄基保护基，并随后与C4发生亲核取代反应得到C5；
    (iii4)C5经溴化得到C6；
    (iii5)C6与C7发生Suzuki偶联得到C8；
    (iii6)C8脱除保护基后与C9发生亲核取代反应得到C10所示的式I化合物，
    在上式中，R 9存在或不存在；
    各式中R 3、R 4、R 5、R 6、R 7、R 8、R 9、m、o、p、q、环Y如权利要求1所述；
    W，X分别为离去基团，各自独立地选自：卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基；PG为保护基团，选自：叔丁氧羰基、苄氧羰基、芴甲氧羰基、烯丙氧羰基、三氟乙酰基、苄基、对甲氧基苄基、三苯甲基；
    或者所述制备方法包括以下步骤：
    Figure PCTCN2022134097-appb-100016
    (iii1)中间体int1与D1发生亲核加成消除反应得到D2；
    (iii2)D2经过还原得到D3；
    (iii3)D3与D4发生偶联反应得到D5；
    (iii4)D5经溴化得到D6；
    (iii5)D6与D7发生Suzuki偶联得到D8；
    (iii6)D8脱除保护基后与D9发生亲核取代反应得到D10所示的式I化合物，
    在上式中，R 9存在或不存在；
    各式中R 3、R 4、R 5、R 6、R 7、R 8、R 9、Z 1、m、o、p、q、环Y如权利要求1所述；
    W，X分别为离去基团，各自独立地选自：卤素、三氟甲磺酸酯基、甲磺酸酯基、对甲苯磺酸酯基；PG为保护基团，选自：叔丁氧羰基、苄氧羰基、芴甲氧羰基、烯丙氧羰基、三氟乙酰基、苄基、对甲氧基苄基、三苯甲基。
  10. 一种药物组合物，其特征在于，所述药物组合物包含：
    权利要求1所述的化合物，或其前药、对映异构体、非对映异构体、外消旋体，或其药学上可接受的盐；和
    药学上可接受的载体。
  11. 如权利要求1所述的化合物，或其前药、对映异构体、非对映异构体、外消旋体，或其药学上可接受的盐或权利要求10所述的药物组合物的用途，其特征在于，用于制备治疗和/或预防与雌激素受体相关的疾病的药物。
  12. 如权利要求11所述的用途，其特征在于，所述与雌激素受体相关的疾病选自：癌症，骨质疏松症，神经退行性疾病，心血管疾病，红斑狼疮，子宫内膜异位症及肥胖症。
PCT/CN2022/134097 2021-12-17 2022-11-24 苯并噻吩类衍生物、及其制备方法和用途 WO2023109461A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202111555326.7A CN116265458A (zh) 2021-12-17 2021-12-17 苯并噻吩类衍生物、及其制备方法和用途
CN202111555326.7 2021-12-17

Publications (1)

Publication Number Publication Date
WO2023109461A1 true WO2023109461A1 (zh) 2023-06-22

Family

ID=86743867

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2022/134097 WO2023109461A1 (zh) 2021-12-17 2022-11-24 苯并噻吩类衍生物、及其制备方法和用途

Country Status (2)

Country Link
CN (1) CN116265458A (zh)
WO (1) WO2023109461A1 (zh)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105008343A (zh) * 2013-02-19 2015-10-28 诺华股份有限公司 作为选择性雌激素受体降解剂的苯并噻吩衍生物及其组合物
CN107406424A (zh) * 2014-12-18 2017-11-28 豪夫迈·罗氏有限公司 雌激素受体调节剂及其用途
CN112839646A (zh) * 2018-08-16 2021-05-25 G1治疗公司 用于治疗医学疾病的苯并噻吩雌激素受体调节剂

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105008343A (zh) * 2013-02-19 2015-10-28 诺华股份有限公司 作为选择性雌激素受体降解剂的苯并噻吩衍生物及其组合物
CN107406424A (zh) * 2014-12-18 2017-11-28 豪夫迈·罗氏有限公司 雌激素受体调节剂及其用途
CN112839646A (zh) * 2018-08-16 2021-05-25 G1治疗公司 用于治疗医学疾病的苯并噻吩雌激素受体调节剂

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
BAENZIGER MARKUS, BAIERL MARCEL, DEVANATHAN KRISHNASWAMY, ESWARAN SUMESH, FU PENG, GSCHWEND BJOERN, HALLER MICHAEL, KASINATHAN GOP: "Synthesis Development of the Selective Estrogen Receptor Degrader (SERD) LSZ102 from a Suzuki Coupling to a C–H Activation Strategy", ORGANIC PROCESS RESEARCH & DEVELOPMENT, AMERICAN CHEMICAL SOCIETY, US, vol. 24, no. 8, 21 August 2020 (2020-08-21), US , pages 1405 - 1419, XP093073195, ISSN: 1083-6160, DOI: 10.1021/acs.oprd.0c00076 *
PARMENTIER MICHAEL, WAGNER MONA, WICKENDICK REGINA, BAENZIGER MARKUS, LANGLOIS AUDREY, GALLOU FABRICE: "A General Kilogram Scale Protocol for Suzuki–Miyaura Cross-Coupling in Water with TPGS-750-M Surfactant", ORGANIC PROCESS RESEARCH & DEVELOPMENT, AMERICAN CHEMICAL SOCIETY, US, vol. 24, no. 8, 21 August 2020 (2020-08-21), US , pages 1536 - 1542, XP093073193, ISSN: 1083-6160, DOI: 10.1021/acs.oprd.0c00281 *
TRIA GEORGE S. ET AL.: "Discovery of LSZ102, a Potent, Orally Bioavailable Selective Estrogen Receptor Degrader (SERD) for the Treatment of Estrogen Receptor Positive Breast Cancer", JOURNAL OF MEDICINAL CHEMISTRY, vol. 61, no. 7, 22 March 2018 (2018-03-22), pages 2837 - 2864, XP055564421, DOI: 10.1021/acs.jmedchem.7b01682 *

Also Published As

Publication number Publication date
CN116265458A (zh) 2023-06-20

Similar Documents

Publication Publication Date Title
CN108884079B (zh) 作为雌激素受体调节剂的6，7-二氢-5h-苯并[7]轮烯衍生物
KR101954322B1 (ko) Ｃ－ｍｅｔ 조절제를 사용하는 방법
CN107485612B (zh) 咪唑并吡啶化合物及其用途
WO2021218110A1 (zh) 一类苯并噻唑基联芳基类化合物、制备方法和用途
AU2014273618B2 (en) Imidazole diketone compound and use thereof
EP3312184A1 (en) Piperidine derivative and preparation method and pharmaceutical use thereof
WO2008001859A1 (fr) AGONISTE DES RÉCEPTEURS δ DES OPIOÏDES
CN112390796A (zh) Kras g12c抑制剂及其在医药上的应用
TW201925204A (zh) 稠合三環化合物
WO2023016484A1 (zh) 磺酰胺衍生物、其制备方法及其在医药上的应用
JP2022515890A (ja) エストロゲン受容体タンパク質分解薬
EP4228757A1 (en) Substituted 6,7-dihydro-5h-benzo[7]annulene compounds and their derivatives, processes for their preparation and therapeutic uses thereof
WO2023109461A1 (zh) 苯并噻吩类衍生物、及其制备方法和用途
WO2022161166A1 (zh) 靶向嵌合化合物、含其的药物组合物及其制备方法和用途
JP6626001B2 (ja) ピリジン誘導体
WO2024078488A1 (zh) 二氢苯并噻喃类衍生物及其制备方法和用途
WO2024042163A1 (en) Novel substituted 2,3,4,9-tetrahydro-1h-pyrido[3,4-b]indole carboxylic acid derivatives, processes for their preparation and therapeutic uses thereof
WO2023198903A1 (en) Novel substituted fluorinated n-propyl-pyrrolidine and n‑propyl-azetidine compounds, processes for their preparation and therapeutic uses thereof
WO2024042185A1 (en) Novel substituted tetrahydroisoquinoline-6- carboxylic acid derivatives, processes for their preparation and therapeutic uses thereof.
WO2022237904A1 (zh) 一种含萘环的化合物、含其的药物组合物及其应用
WO2022174796A1 (zh) 作为Wee-1抑制剂的嘧啶化合物
WO2022166980A1 (zh) 杂芳基并哌啶类衍生物及其药物组合物和应用
WO2024042157A1 (en) Novel substituted quinoline and tetrahydronaphthalene carboxylic acid derivatives and therapeutic uses thereof
CA3223859A1 (en) Salt and crystal forms of an epidermal growth factor receptor inhibitor
WO2024042187A1 (en) Novel substituted benzothiophene-6-carboxylic acid derivatives, processes for their preparation and therapeutic uses thereof

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 22906212

Country of ref document: EP

Kind code of ref document: A1