WO2021098817A1 - Dioxopiperazine derivative, preparation method therefore and pharmaceutical use thereof - Google Patents

Dioxopiperazine derivative, preparation method therefore and pharmaceutical use thereof Download PDF

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Publication number
WO2021098817A1
WO2021098817A1 PCT/CN2020/130361 CN2020130361W WO2021098817A1 WO 2021098817 A1 WO2021098817 A1 WO 2021098817A1 CN 2020130361 W CN2020130361 W CN 2020130361W WO 2021098817 A1 WO2021098817 A1 WO 2021098817A1
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Prior art keywords
phenyl
chloro
mmol
tert
tetrazol
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PCT/CN2020/130361
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French (fr)
Chinese (zh)
Inventor
吴俊军
陆银锁
肖瑛
王延彬
吕洋
邢伟
王汝欢
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深圳信立泰药业股份有限公司
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Priority to CN202080079337.XA priority Critical patent/CN114728914B/en
Publication of WO2021098817A1 publication Critical patent/WO2021098817A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
    • C07D241/04—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having no double bonds between ring members or between ring members and non-ring members

Definitions

  • the invention belongs to the technical field of chemical medicines, and relates to dioxopiperazine derivatives, a preparation method thereof and their application in medicine.
  • the present invention provides a compound of formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein all variables are as defined herein.
  • These compounds are inhibitors of selective factor XIa (Factor XIa, FXIa for short).
  • the present invention also relates to a pharmaceutical composition containing these compounds and the use of the compounds in the treatment of diseases such as thromboembolism.
  • Cardiovascular and cerebrovascular diseases such as cerebrovascular, cerebral infarction, myocardial infarction, coronary heart disease, and arteriosclerosis take the lives of nearly 12 million people worldwide each year, which is close to a quarter of the world's total deaths, becoming the number one enemy of human health. In China, more than 2.6 million people die from cardiovascular disease each year, and 75% of the surviving patients are disabled, of which more than 40% are severely disabled. Thrombosis caused by cardiovascular and cerebrovascular diseases and diabetes and its complications has become an urgent problem to be solved today.
  • the human blood coagulation process is composed of intrinsic pathways, extrinsic pathways and common pathways (Annu.Rev.Med.2011.62:41–57). It is caused by the sequential activation of multiple zymogens. A chain reaction in which the process continues to be strengthened and amplified.
  • the coagulation cascade is initiated by the endogenous pathway (also called the contact activation pathway) and the exogenous pathway (also called the tissue factor pathway) to generate FXa, and then through the common pathway to generate thrombin (FIIa), and finally form fibrin.
  • the endogenous pathway refers to the process by which factor XII is activated to form the XIa-VIIIa-Ca 2+ -PL complex and activate factor X.
  • the exogenous coagulation pathway is the release of tissue factor (TF) to TF-VIIa-
  • the common pathway refers to the process in which the two pathways are combined into one after the formation of factor Xa, which activates prothrombin and finally generates fibrin.
  • FXI is necessary to maintain the endogenous pathway and is in the process of amplification of the coagulation cascade. Play a key role.
  • FXIa activated FXI
  • FXIa is currently an emerging target for inhibiting thrombosis.
  • Patent applications that disclose compounds with FXIa inhibitory activity include WO9630396, WO9941276, WO2013093484, WO2004002405, WO2013056060, WO2017005725, WO2017/023992, WO2018041122, etc.
  • Bayer's antisense oligonucleotide BAY-2306001 has entered the phase II clinical study.
  • the compounds of the present invention have higher activity.
  • the compound of the present invention exhibits excellent anticoagulant effects on human blood, and has good pharmacokinetic activity, and can be used to effectively treat and/or prevent cardiovascular and cerebrovascular diseases and thrombotic symptoms.
  • the invention provides a series of oxopyridazinamide derivatives, their preparation methods and their medical applications.
  • the present invention provides a compound of formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein all variables are as defined herein.
  • the present invention also relates to a pharmaceutical composition containing these compounds and the use of the compounds in the treatment of diseases such as thromboembolism.
  • the present invention is implemented through the following technical solutions:
  • R 1 is selected from R 3 substituted or unsubstituted tetrazole, R 3 substituted or unsubstituted triazole;
  • R 2 is selected from R 4 substituted or unsubstituted benzene ring, wherein R 4 is selected from -NR 5 -(CH 2 )n-CO-(CH 2 )n-NR 6 R 7 , -NR 5 -SO 2 -NR 6 R 7 ;
  • Ar is selected from at least one of the following groups substituted or unsubstituted by R 8:
  • R 3 is selected from hydrogen, halogen, C 1-4 alkyl, halogen substituted C 1-4 alkyl;
  • R 5 , R 6 , and R 7 are independently selected from hydrogen, C 1-4 alkyl, C 1-4 alkoxy -C 1-4 alkyl, -SO 2 -C 1-4 alkane, -SO 2 -Benzene, -C 1-6 mono or dihydric alcohol, -(CH 2 )nC 3-12 aliphatic ring, or wherein any one or more of NR 5 and NR 6 R 7 form a ring through -(CH 2 )n-; or NR 6 R 7 together form C 3-12
  • R 8 is selected from hydrogen, halogen, C 1-4 alkyl, hydroxyl, -C 1-4 carboxylic acid, -C 1-4 carboxylic acid-C 1-4 alcohol ester;
  • R 9 is selected from hydrogen, -(CH 2 )n-OH, -SO 2 -C 1-4 alkane, -(CH 2 )n-COOH, -amide, cyano, NR 10 R 11 -C 1-4 Alkoxy, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 alkoxy-C 1-4 alkyl, -CO-morpholine, -CO-NR 12 -(CH 2 ) n-OH, HOOC-C 1-4 alkoxy,
  • R 10 , R 11 , and R 12 are independently selected from hydrogen or C 1-4 alkyl
  • n 0-6.
  • the -NR 5 -(CH 2 )n-CO-(CH 2 )n-NR 6 R 7 includes Wherein, R 7 and R 9 are as defined above.
  • the C 1-4 alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl;
  • -C 1-4 carboxylic acid is selected from formic acid, acetic acid, propionic acid, n-butyric acid, isobutyric acid, tert-butyric acid;
  • -C 1-4 carboxylic acid-C 1-4 alcohol ester is selected from methyl formate, ethyl formate, propyl formate, butyl formate, methyl acetate, ethyl acetate, propyl acetate, butyl acetate, propyl Methyl acid, ethyl propionate, propyl propionate, butyl propionate, methyl butyrate, ethyl butyrate, propyl butyrate, butyl butyrate.
  • the halogen is selected from fluorine, chlorine, bromine, and iodine.
  • the C 1-4 alkane is selected from methane, ethane, propane, isopropane, n-butane, isobutane, sec-butane, and tert-butane;
  • the C 1-4 alkoxy group is selected from methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy;
  • the -(CH 2 )n-OH is selected from hydroxyl, methanol, ethanol, n-propanol, and n-butanol;
  • the -C 1-6 monohydric or dihydric alcohol is selected from methanol, ethanol, n-propanol, n-butanol, tert-butanol, 1,3-butanediol, 3-methylbutan-1-ol, 3 -Methylpentan-1-ol, 4-methylpentan-1ol, 3-methylhexan-1ol, 4-methylhexan-1ol;
  • the alkoxy group of the NR 10 R 11 -C 1-4 is selected from the group consisting of methylamine methoxy, ethylamine methoxy, propylamine methoxy, butylamine methoxy, methylamine ethoxy, and ethylamine ethoxy.
  • the C 1-4 alkoxy-C 1-4 alkyl group is selected from methoxymethyl, ethoxymethyl, propoxymethyl, butoxymethyl, methoxyethyl, ethoxyethyl, propoxy Ethyl, butoxyethyl, methoxypropyl, ethoxypropyl, propoxypropyl, butoxypropyl, methoxybutyl, ethoxybutyl, propoxybutyl, butoxybutyl;
  • the -CO-NR 12 -(CH 2 )n-OH is selected from -CO-NH-CH 2 OH, -CO-N(CH 3 )-CH 2 OH, -CO-NH-CH 2 CH 2 OH, -CO-N(CH 3 )-CH 2 CH 2 OH;
  • the HOOC-C 1-4 alkoxy group is selected from HOOC-methoxy, HOOC-ethoxy, HOOC-propoxy, HOOC-isopropoxy, HOOC-n-butoxy, HOOC-isobutoxy Group, HOOC-sec-butoxy, HOOC-tert-butoxy.
  • the C 3-12 aliphatic ring is selected from cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, and cyclodecane.
  • More than one carbon atom of the C 3-12 fat is replaced by 0-2 N, O, S atoms, selected from:
  • n 0,1,2,3,4,5,6.
  • R 1 is selected from tetrazolium and triazole
  • R 2 is selected from benzene ring
  • Ar is selected from the following groups:
  • the compound or a pharmaceutically acceptable salt thereof is selected from the following compounds:
  • the pharmaceutically acceptable salt refers to a compound prepared with a pharmaceutically acceptable acid or base.
  • more than one hydrogen atom of the compound is replaced by an isotope deuterium.
  • Another object of the present invention is to provide a pharmaceutical composition
  • a pharmaceutical composition comprising the aforementioned compound of formula (I), or its stereoisomers, tautomers, pharmaceutically acceptable salts and more than one pharmaceutically acceptable a.
  • Another object of the present invention is to provide the compound of formula (I), or its stereoisomers, tautomers, pharmaceutically acceptable salts, and containing the compound, or its stereoisomers,
  • the pharmaceutical composition of tautomers and pharmaceutically acceptable salts is used in the preparation of pharmaceuticals for the treatment of FXIa-related diseases, in particular, the pharmaceutical uses related to thrombosis-related diseases.
  • pharmaceutically acceptable salt refers to a salt of the compound of the present invention, which is prepared from the compound with specific substituents discovered in the present invention and a pharmaceutically acceptable acid or base.
  • the compounds provided by the present invention also exist in prodrug forms.
  • the prodrugs of the compounds described herein easily undergo chemical changes under physiological conditions to transform into the compounds of the invention.
  • prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in the in vivo environment.
  • Certain compounds of the present invention may exist in unsolvated or solvated forms, including hydrated forms.
  • the solvated form is equivalent to the unsolvated form, and both are included in the scope of the present invention.
  • the compounds of the present invention may exist in specific geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers Conformers, (D)-isomers, (L)-isomers, and their racemic mixtures and other mixtures, such as enantiomers or diastereomer-enriched mixtures, all of these mixtures belong to Within the scope of the present invention.
  • Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All these isomers and their mixtures are included in the scope of the present invention.
  • optically active (R)- and (S)-isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If you want to obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with chiral auxiliary agents, in which the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure The desired enantiomer.
  • the molecule when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), it forms a diastereomeric salt with an appropriate optically active acid or base, and then passes through a conventional method known in the art The diastereoisomers are resolved, and then the pure enantiomers are recovered.
  • the separation of enantiomers and diastereomers is usually accomplished through the use of chromatography, which uses a chiral stationary phase and is optionally combined with chemical derivatization (for example, the formation of amino groups from amines). Formate).
  • the atoms of the compound molecules of the present invention are isotopes, and isotope derivatization can generally extend the half-life, reduce the clearance rate, enhance the stability of metabolism, and increase the activity in the body. And, an embodiment is included in which at least one atom is substituted with atoms having the same atomic number (number of protons) and different mass numbers (sum of protons and neutrons).
  • isotopes included in the compounds of the present invention include hydrogen atoms, carbon atoms, nitrogen atoms, oxygen atoms, phosphorus atoms, sulfur atoms, fluorine atoms, and chlorine atoms, which respectively include 2 H, 3 H, 13 C, 14 C, and 15 N, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 Cl.
  • radioisotopes that emit radiation as they decay such as 3 H or 14 C, can be used for topographical examinations of pharmaceutical preparations or compounds in the body.
  • the stable isotope neither decays or changes with its amount, nor is it radioactive, so it can be used safely.
  • the isotopes can be converted according to general methods by replacing the reagents used in the synthesis with reagents containing the corresponding isotopes.
  • the compound of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms constituting the compound.
  • compounds can be labeled with radioisotopes, such as deuterium ( 2 H), iodine-125 ( 125 I), or C-14 ( 14 C). All changes in the isotopic composition of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.
  • one or more hydrogen atoms of the compound of the present invention are replaced by the isotope deuterium ( 2 H).
  • the compound of the present invention has the effects of prolonging half-life, reducing clearance, enhancing metabolic stability, and improving in vivo activity.
  • the preparation method of the isotope derivative usually includes a phase transfer catalysis method.
  • the preferred deuteration method uses a phase transfer catalyst (e.g., tetraalkylammonium salt, NBu 4 HSO 4 ).
  • a phase transfer catalyst e.g., tetraalkylammonium salt, NBu 4 HSO 4 .
  • the use of a phase transfer catalyst to exchange the methylene protons of the diphenylmethane compound results in the use of deuterated silanes (e.g. triethyl deuterated monosilane) or Lewis acids such as trichlorosilane in the presence of an acid (e.g., methanesulfonic acid)
  • Aluminum chloride is reduced with deuterated sodium borate to introduce higher deuterium.
  • pharmaceutically acceptable carrier refers to any preparation carrier or medium that can deliver an effective amount of the active substance of the present invention, does not interfere with the biological activity of the active substance, and has no toxic or side effects to the host or patient.
  • Representative carriers include water and oil. , Vegetables and minerals, cream base, lotion base, ointment base, etc. These bases include suspending agents, tackifiers, penetration enhancers and the like. Their formulations are well known to those skilled in the field of cosmetics or topical medicine. For other information about the carrier, you can refer to Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), and the content of this document is incorporated herein by reference.
  • excipient generally refers to the carrier, diluent and/or medium required to formulate an effective pharmaceutical composition.
  • the term "effective amount” or “therapeutically effective amount” refers to a sufficient amount of a drug or agent that is non-toxic but can achieve the desired effect.
  • the "effective amount” of one active substance in the composition refers to the amount required to achieve the desired effect when combined with another active substance in the composition.
  • the determination of the effective amount varies from person to person, and depends on the age and general conditions of the recipient, as well as the specific active substance. The appropriate effective amount in a case can be determined by those skilled in the art according to routine experiments.
  • active ingredient refers to a chemical entity that can effectively treat the target disorder, disease or condition.
  • the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and those well known to those skilled in the art Equivalent alternatives, preferred implementations include but are not limited to the embodiments of the present invention.
  • the structure of the compound is determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS).
  • NMR shift ( ⁇ ) is given in units of 10-6 (ppm).
  • NMR was measured with Bruker AVANCE-III nuclear magnetometer, the solvent was deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and the internal standard was tetramethylsilane (TMS).
  • MS is measured with ISQ EC mass spectrometer (manufacturer: Thermo, model: ISQ EC).
  • HPLC high performance liquid chromatography
  • CombiFlash rapid preparation instrument uses CombiFlash Rf+LUMEN (TELEDYNE ISCO).
  • the thin layer chromatography silica gel plate uses Yantai Yinlong HSGF254 or GF254 silica gel plate.
  • the size of the silica gel plate used in thin layer chromatography (TLC) is 0.17mm ⁇ 0.23mm, and the size of thin layer chromatography separation and purification products is 0.4mm. ⁇ 0.5mm.
  • the silica gel column chromatography generally uses Rushan Shangbang silica gel 100-200 mesh silica gel as the carrier.
  • Step B Synthesis of (2-((5-chloro-2-nitrophenyl)amino)-2-oxoacetyl)-L-phenylalanine tert-butyl ester
  • Step C Synthesis of (S)-2-(4-(5-chloro-2-nitrophenyl)-2,3-dioxopiperazin-1-yl)-3-phenylpropionic acid tert-butyl ester
  • Step D Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-phenylpropionic acid tert-butyl ester
  • Step E Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Tert-butyl phenylpropionate
  • Step F Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionic acid
  • Step G Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N -(2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-3-phenylpropionamide
  • the reaction was quenched by adding water, a solid was precipitated, filtered, and a filter cake was used to obtain 70 mg of crude product.
  • the crude product was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm.
  • Step A Synthesis of (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido) tert-butyl benzoate
  • Step B Synthesis of (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)benzoic acid
  • the reaction liquid was distilled under reduced pressure.
  • the obtained residue was purified by preparative high performance liquid chromatography.
  • the separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm.
  • Step A Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N -(3-oxoisoindolin-5-yl)-3-phenylpropionamide
  • the reaction was quenched by adding water, extracted with dichloromethane (10 ml ⁇ 3 times), and distilled under reduced pressure.
  • the residue obtained was purified by preparative high performance liquid chromatography.
  • the separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm.
  • Step A Synthesis of (S)-5-(3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-tert-butyl phenylpropoxy)phenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylic acid tert-butyl ester
  • the reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml ⁇ 3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure.
  • the residue obtained was purified by preparative high performance liquid chromatography.
  • the separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm.
  • Step A Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N -(4-(5-oxo-2-5-dihydro-1H-pyrazol-3-yl)phenyl)-3-phenylpropionamide
  • the reaction liquid was distilled under reduced pressure.
  • the obtained residue was purified by preparative high performance liquid chromatography.
  • the separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm.
  • Step A Synthesis of (S)-3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido) tert-butyl benzoate
  • Step B Synthesis of (S)-3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)benzoic acid
  • the reaction liquid was distilled under reduced pressure.
  • the obtained residue was purified by preparative high performance liquid chromatography.
  • the separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm.
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid tert-butyl ester
  • the reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml ⁇ 3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure.
  • the residue obtained was purified with a TLC plate (ethyl acetate: n-hexane).
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid
  • the reaction liquid was distilled under reduced pressure.
  • the obtained residue was purified by preparative high performance liquid chromatography.
  • the separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm.
  • 20 mg of light yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxo were obtained.
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • the reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml ⁇ 3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure.
  • the residue obtained was purified with a TLC plate (ethyl acetate: n-hexane).
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-indole-2-carboxylic acid
  • the reaction liquid was distilled under reduced pressure.
  • the obtained residue was purified by preparative high performance liquid chromatography.
  • the separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm.
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • the reaction liquid was distilled under reduced pressure.
  • the obtained residue was purified by preparative high performance liquid chromatography.
  • the separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm.
  • chromatographic column X select C18 19mm*150mm
  • mobile phase water (containing 0.05% trifluoroacetic acid) and acetonitrile
  • flow rate 25 ml/min
  • gradient within 7 minutes, acetonitrile rises from 5% to 100%
  • detection wavelength 254nm.
  • Step B Synthesis of 3-chloro-5-nitro-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step C Synthesis of 3-chloro-5-amino-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step D Synthesis of (S)-3-chloro-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine -1-yl)-3-phenylpropionamide)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step E Synthesis of (S)-3-chloro-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine -1-yl)-3-phenylpropionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of 1-methyl-5-nitro-1H-indole-2-carboxylic acid ethyl ester
  • reaction solution was quenched by adding saturated aqueous ammonium chloride solution, the mixed solution was extracted with ethyl acetate (40 ml ⁇ 3 times), the organic phases were combined, and the organic phase was saturated brine (30 ml ⁇ 3 times), anhydrous sodium sulfate Dry and concentrate under reduced pressure.
  • Step C Synthesis of 1-methyl-5-nitro-1H-indole-2-carboxylic acid tert-butyl ester
  • Step D Synthesis of 1-methyl-5-amino-1H-indole-2-carboxylic acid tert-butyl ester
  • Step E Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-nitrophenyl)propionylamino)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester
  • the reaction was quenched by adding water to the reaction solution.
  • the mixture was extracted with ethyl acetate (20 ml ⁇ 3 times).
  • the organic phase was combined, and the organic phase was first saturated brine (10 ml ⁇ 3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure.
  • Step F Synthesis of (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3 -Dioxopiperazin-1-yl)propionylamino)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester
  • Step G Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-(piperidine-1-carboxamido)phenyl)propionamido)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester
  • Step H Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1-methyl-1H-indole-2-carboxylic acid
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Hydroxypiperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step C Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(1,1-dioxotetrahydro-2H-thiopyran-4-yl)ureido)phenyl)propionylamino)-1H-indole-1,2- Di-tert-butyl dicarboxylate
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(1,1-dioxotetrahydro-1H-thiopyran-4-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step B Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionylamino)-1H-indole-1,2 -Di-tert-butyl dicarboxylate
  • Step C Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxy acid
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(Methylsulfonyl)piperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(3-(4-(4-(tert-butoxycarbonyl)piperidine-1-carboxamido)phenyl)-2-(4-(5-chloro-2 -(1H-tetrazole)-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step B Synthesis of (S)-5-(3-(4-(4-carboxypiperidine-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H-tetrazole) -1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(Methylsulfonyl)ureido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-((S)-3-hydroxypiperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step B Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-((4-((S)-3-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of methyl 3-(3-oxopiperazin-1-yl)cyclobutane-1-carboxylate
  • Step C Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(3-(Hydroxymethyl))cyclobutyl)-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
  • Step D Synthesis of (S)-2-(4-(2-amino-5-chloro-phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-( 3-(Hydroxymethyl))cyclobutyl)-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
  • Step E Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(3-(Hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step F Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-(-(3-(hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2 -Di-tert-butyl dicarboxylate
  • Step G Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(3-(hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxy acid
  • Step C Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
  • Step D Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(tetrahydrofuran) -3-yl)methyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
  • Step E Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step F Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester
  • Step G Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(3-(4-(4-(2-(tert-butoxy)-2-oxoethoxy)piperidine-1-carboxamido)phenyl)- 2-(4-(5-)chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole -1,2-Di-tert-butyl dicarboxylic acid
  • Step B Synthesis of (S)-5-(3-(4-(4-(carboxymethoxy)piperidine-1-carboxamido)phenyl)-2-(4-(5-chloro-2- (1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxy Di-tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1, Di-tert-butyl 2-dicarboxylate
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2- carboxylic acid
  • Step A Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(4-Hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl
  • Step B Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2S,3S)-1,3-dihydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
  • Step B Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2R,3R)-1,3-dihydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of tert-butyl (S)-2-amino-3-(4-bromophenyl)propionate
  • Step B Synthesis of (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid tert-butyl ester
  • Piperazine-2-one (11.0 g, 55.5 mmol) was dissolved in a mixed solution of methanol (20.0 mL) and acetic acid (0.5 mL). Subsequently, acetone (6.78 g, 111.0 mmol) and sodium cyanoborohydride (4.19 g, 66.6 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
  • Step D Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionic acid tert Butyl
  • Step E Synthesis of (S)-2-amino-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionic acid tert-butyl ester hydrochloride
  • Step F Synthesis of (S)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido)-3-(4-(4- Isopropyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
  • Step G Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-isopropyl-2-oxopiperazine)-tert-butylazin-1-yl)phenyl)tert-butyl propionate
  • Step H Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-isopropyl 2-oxopiperazin-1-yl) phenyl) tert-butyl propionate
  • Step I Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step J Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionic acid
  • Step K Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step L Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-3-(4-bromophenyl)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido ) Tert-butyl propionate
  • Step B Synthesis of (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazine -1-yl) tert-butyl propionate
  • Piperazine-2-one (1.0 g, 9.9 mmol) was dissolved in a mixed solution of methanol (10.0 mL) and acetic acid (0.2 mL). Subsequently, 4-methylcyclohexan-1-one (2.6 g, 19.9 mmol) and sodium cyanoborohydride (1.3 g, 19.9 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
  • Step D Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(4-Methoxycyclohexyl))-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
  • N,N-dimethylethyl-1,2-diamine (292 mg, 3.22 mmol)
  • cuprous iodide (315 mg, 1.66 mmol)
  • cesium carbonate (1 g, 3.22 mmol).
  • Step E Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(4 -Methoxycyclohexyl)-2-oxopiperazin)-1-yl)phenyl)tert-butyl propionate
  • Step F Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step G Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-(methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid
  • Step H Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2-dicarboxy Di-tert-butyl ester
  • Triazole N,N,N',N'-tetramethylurea hexafluorophosphate (245 mg, 0.64 mmol), N,N-diisopropylethylamine (111 mg, 0.86 mmol) , Stir at room temperature for 18 hours.
  • Step I Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Piperazine-2-one (2.0 g, 20.0 mmol) was dissolved in acetonitrile (20.0 mL). Subsequently, 1-bromo-2-methoxyethane (5.6 g, 40.0 mmol) and potassium carbonate (5.5 g, 40.0 mmol) were added to the above solution. Stir overnight at 100 degrees Celsius.
  • Step B Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(2-Methoxyethyl)-)2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
  • Step C Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(2 -Methoxyethyl)-2-oxopiperazin)-1-yl)phenyl)tert-butyl propionate
  • Step D Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step E Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(2-(Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionic acid
  • Step F Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-1,2-dicarboxy Di-tert-butyl ester
  • Triazole N,N,N',N'-tetramethylurea hexafluorophosphate (171 mg, 0.45 mmol), N,N-diisopropylethylamine (77 mg, 0.60 mmol) , Stir at room temperature for 18 hours.
  • Step G Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Piperazine-2-one 500 mg, 2.5 mmol was dissolved in a mixed solution of methanol (10.0 mL) and acetic acid (0.2 mL). Subsequently, tetrahydro-4H-pyran-4-one (788 mg, 5.0 mmol) and sodium cyanoborohydride (317 mg, 5.0 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
  • Step B Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl) tert-butyl propionate
  • N,N-dimethylethyl-1,2-diamine (292 mg, 3.3 mmol), cuprous iodide (315 mg, 1.6 mmol) and cesium carbonate (1.08 g, 3.3 mmol). Stir at 110 degrees Celsius for 18 hours.
  • Step C Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2-oxo -4-(Tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)tert-butyl propionate
  • Step D Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)tert-butyl propionate
  • Step E Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionic acid
  • Step F Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-1, Di-tert-butyl 2-dicarboxylate
  • Step G Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2- carboxylic acid
  • Piperazine-2-one (1.5 g, 7.5 mmol) was dissolved in a mixed solution of methanol (20.0 mL) and acetic acid (0.4 mL). Subsequently, 4-hydroxycyclohexane-1-one (1.7 g, 15.0 mmol) and sodium cyanoborohydride (951 mg, 15.0 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
  • Step B Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(4-Hydroxycyclohexyl))-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
  • N,N-dimethylethane-1,2-diamine (175 mg, 1.99 mmol)
  • cuprous iodide 189 mg, 0.99 mmol
  • cesium carbonate 645 mg, 1.99 mmol
  • Step C Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(4 -Hydroxycyclohexyl)-2-oxopiperazin)-1-yl)phenyl)tert-butyl propionate
  • Step D Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step E Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid
  • Step F Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step G Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid compound
  • Piperazine-2-one 500 mg, 4.9 mmol was dissolved in a mixed solution of methanol (10.0 mL) and acetic acid (0.2 mL). Subsequently, dihydro-2H-pyran-3(4H)-one (1.0 g, 10.0 mmol) and sodium cyanoborohydride (627 mg, 10.0 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
  • Step B Synthesis of (2S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(2-oxo-4-(tetrahydro)-2H-pyran-3-yl)piperazin-1-yl)phenyl)tert-butyl propionate
  • N,N-dimethylethane-1,2-diamine (175 mg, 1.99 mmol)
  • cuprous iodide 189 mg, 0.99 mmol
  • cesium carbonate 645 mg, 1.99 mmol
  • Step C Synthesis of (2S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2-oxo -4-(Tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)tert-butyl propionate
  • Step D Synthesis of (2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl) tert-butyl propionate
  • (2S)-2-(4-(2-Amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2-oxo-4- (Tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)tert-butyl propionate (260 mg, 0.4 mmol) and triethyl orthoformate (430 mg, 2.9 mmol) Dissolve in acetic acid (5.0 ml). Subsequently, sodium azide (188 mg, 2.9 mmol) was added to the above solution. Stir at 70 degrees Celsius for 1 hour.
  • Step E Synthesis of (2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionic acid
  • Step F Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2- Tert-butyl carboxylate
  • Step G Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2- carboxylic acid
  • Step A Synthesis of 6-nitro-1H-indole-2-carboxylic acid tert-butyl ester
  • Step B Synthesis of 6-amino-1H-indole-2-carboxylic acid tert-butyl ester
  • Step C Synthesis of (S)-6-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step D Synthesis of (S)-6-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxide Piperazin-1-yl) tert-butyl propionate
  • Step B Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((phenoxycarbonyl)yl)amino)phenyl)tert-butyl propionate
  • Step C Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Cyanopiperidine-1-carboxamido)phenyl) tert-butyl propionate
  • Step D Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionic acid
  • Step E Synthesis of (S)-N-(4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-((4-hydroxy-2-oxo-1,2-dihydroquinolin-6-yl)amino)-3-oxopropyl)phenyl)-4-cyanopiper Pyridine-1-carboxamide
  • Step A Synthesis of tert-butyl (S)-2-amino-3-(4-nitrophenyl)propionate
  • Step B Synthesis of tert-butyl (S)-2-(2-methoxy-2-oxoacetamido)-3-(4-nitrophenyl)propionate
  • Step C Synthesis of (S)-2-((1-(tert-butoxy)-3-(4-nitrophenyl)-1-oxopropan-2-yl)amino)-2-oxoacetic acid
  • Step E Synthesis of N 1 ,N 1 -diallyl-4-chlorobenzene-1,2-diamine
  • N,N-diallyl-4-chloro-2-nitroaniline (27.5 g, 108.8 mmol) to ethyl acetate, and add stannous chloride dihydrate in batches under ice bath (122.8 g, 544.0 mmol), under N 2 protection, react at room temperature overnight.
  • Step F Synthesis of (S)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido)-3-(4-nitrobenzene Base) tert-butyl propionate
  • Step G Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -Nitrophenyl) tert-butyl propionate
  • the pad was suction filtered with anhydrous sodium sulfate, the filter cake was washed with 200 ml of ethyl acetate, the filtrate was extracted with ethyl acetate (100 ml ⁇ 3 times), the organic phases were combined, and the organic phase was first used with saturated brine (100 ml ⁇ 2 times) washing, then drying with anhydrous sodium sulfate, and finally concentrating under reduced pressure.
  • Step H Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-nitrophenyl) Tert-butyl propionate
  • Step I Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-Nitrophenyl) tert-butyl propionate
  • Step J Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-Nitrophenyl)propionic acid
  • Step K Synthesis of 5-nitro-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step L Synthesis of 5-amino-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step M Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-nitrophenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step N Synthesis of (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3 -Dioxopiperazin-1-yl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step O Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step P Synthesis of 5-((2S)-3-(4-(3-((1,4-dioxan-2-yl)methyl)ureido)phenyl)-2-(4-(5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester
  • Step Q Synthesis of 5-((2S)-3-(4-(3-((1,4-dioxan-2-yl)methyl)ureido)phenyl)-2-(4-(5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step B Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
  • Step B Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
  • Step B Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
  • Step B Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((3-Methyloxetan-3-yl)methyl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
  • Step B Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((3-Methyloxetan-3-yl)methyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid

Abstract

Provided are a dioxopiperazine derivative, a preparation method therefor and pharmaceutical use thereof. In particular, provided are compounds of Formula (I) or a stereoisomer, a tautomer, a pharmaceutically acceptable salt thereof. The compounds are inhibitors of the selective factor XIa (Factor XIa, abbreviated to FXIa). The present invention also relates to a pharmaceutical composition comprising the compounds and use of the compounds in drugs for treating diseases such as thromboembolism.

Description

二氧代哌嗪类衍生物、其制备方法及其在医药上的应用Dioxopiperazine derivatives, preparation method thereof and application in medicine 技术领域Technical field
本发明属于化学药物技术领域，涉及二氧代哌嗪类衍生物、其制备方法及其在医药上的应用。具体而言，本发明提供式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中所有变量如本文所定义。这些化合物是选择性因子XIa(Factor XIa，简称FXIa)的抑制剂。本发明还涉及包含这些化合物的药物组合物以及使用该化合物治疗血栓栓塞等疾病的药物中的用途。The invention belongs to the technical field of chemical medicines, and relates to dioxopiperazine derivatives, a preparation method thereof and their application in medicine. Specifically, the present invention provides a compound of formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein all variables are as defined herein. These compounds are inhibitors of selective factor XIa (Factor XIa, FXIa for short). The present invention also relates to a pharmaceutical composition containing these compounds and the use of the compounds in the treatment of diseases such as thromboembolism.
背景技术Background technique
全球每年脑血管、脑梗塞、心肌梗塞、冠心病、动脉硬化等心脑血管疾病夺走近1200万人的生命，接近世界总死亡人数的1/4，成为人类健康的头号大敌。中国每年死于心血管疾病的人数达到260万人以上，存活的患者75％致残，其中40％以上重残。由心脑血管疾病和糖尿病及其并发症引起的血栓问题，成为当今要解决的刻不容缓的问题。Cardiovascular and cerebrovascular diseases such as cerebrovascular, cerebral infarction, myocardial infarction, coronary heart disease, and arteriosclerosis take the lives of nearly 12 million people worldwide each year, which is close to a quarter of the world's total deaths, becoming the number one enemy of human health. In China, more than 2.6 million people die from cardiovascular disease each year, and 75% of the surviving patients are disabled, of which more than 40% are severely disabled. Thrombosis caused by cardiovascular and cerebrovascular diseases and diabetes and its complications has become an urgent problem to be solved today.
人体血液凝固过程由内源性途径(intrinsic pathway)、外源性途径(extrinsic pathway)和共同通路组成(Annu.Rev.Med.2011.62:41–57)，是通过多种酶原被顺序激活而过程不断得到加强和放大的一种连锁反应。凝血级联反应由内源性途径(又称接触激活途径)及外源性途径(又称组织因子途径)启动生成FXa，再经共同途径生成凝血酶(FIIa)，最终形成纤维蛋白。The human blood coagulation process is composed of intrinsic pathways, extrinsic pathways and common pathways (Annu.Rev.Med.2011.62:41–57). It is caused by the sequential activation of multiple zymogens. A chain reaction in which the process continues to be strengthened and amplified. The coagulation cascade is initiated by the endogenous pathway (also called the contact activation pathway) and the exogenous pathway (also called the tissue factor pathway) to generate FXa, and then through the common pathway to generate thrombin (FIIa), and finally form fibrin.
内源性途径是指由XII因子被激活形XIa-VIIIa-Ca 2+-P L复合物、并激活X因子的过程,外源性凝血途径则是从组织因子(TF)释放到TF-VIIa-Ca 2+复合物形成并激活因子Ⅹ的过程。共同通路是指因子Xa形成后,两条途径合二为一,激活凝血酶原并最终生成纤维蛋白的过程，其中FXI是维持内源性途径所必需的，而且在凝血级联反应放大过程中发挥关键作用。在凝血级联反应中，凝血酶可反馈激活FXI，活化的FXI(FXIa)又促使凝血酶的大量产生，从而使凝血级联反应放大。因此，FXI的拮抗剂被广泛开发，用于各种血栓的治疗。 The endogenous pathway refers to the process by which factor XII is activated to form the XIa-VIIIa-Ca 2+ -PL complex and activate factor X. The exogenous coagulation pathway is the release of tissue factor (TF) to TF-VIIa- The process by which the Ca 2+ complex forms and activates factor X. The common pathway refers to the process in which the two pathways are combined into one after the formation of factor Xa, which activates prothrombin and finally generates fibrin. Among them, FXI is necessary to maintain the endogenous pathway and is in the process of amplification of the coagulation cascade. Play a key role. In the coagulation cascade, thrombin can feedback activation of FXI, and activated FXI (FXIa) promotes the production of thrombin in large quantities, thereby amplifying the coagulation cascade. Therefore, FXI antagonists have been widely developed for the treatment of various thrombosis.
传统的抗凝药物，如华法林、肝素、低分子量肝素(LMWH)，以及近年上市的新药，如FXa抑制剂(利伐沙班、阿哌沙班等)和凝血酶抑制剂(达比加群酯、水蛭素等)，对减少血栓形成均具有较好效果，以其显著有效性占据广大心脑血管市场，然而其副作用也越来越显著，其中“出血风险(bleeding risk)”是首当其冲最为严峻的问题之一(N Engl J Med1991；325:153-8、Blood.2003；101:4783-4788)。Traditional anticoagulant drugs, such as warfarin, heparin, low molecular weight heparin (LMWH), and new drugs marketed in recent years, such as FXa inhibitors (rivaroxaban, apixaban, etc.) and thrombin inhibitors (darby Gatran etexilate, hirudin, etc.) have good effects on reducing thrombosis, occupying the vast cardiovascular and cerebrovascular market with its significant effectiveness, but its side effects are becoming more and more significant. Among them, the "bleeding risk" is Bear the brunt of one of the most serious problems (N Engl J Med1991; 325:153-8, Blood. 2003; 101:4783-4788).
研究发现，在血栓模型中，抑制FXIa因子可以有效抑制血栓的形成，但在更为严重的血栓情况下，FXIa的作用微乎其微(Blood.2010；116(19):3981-3989)。临床统计显示，提高FXIa的量会增加VTE的患病率(Blood 2009；114:2878-2883)，而FXIa严重不足者其患有DVT的风险性减少(Thromb Haemost 2011；105:269–273)。Studies have found that in the thrombosis model, inhibiting FXIa factor can effectively inhibit the formation of thrombus, but in the case of more severe thrombosis, the effect of FXIa is minimal (Blood. 2010; 116(19): 3981-3989). Clinical statistics show that increasing the amount of FXIa will increase the prevalence of VTE (Blood 2009; 114: 2878-2883), and people with severe FXIa deficiency have a reduced risk of DVT (Thromb Haemost 2011; 105: 269–273) .
FXIa作为目前抑制血栓的新兴靶点，公开具有FXIa抑制活性的化合物的专利申请有WO9630396、WO9941276、WO2013093484、WO2004002405、WO2013056060、WO2017005725、WO2017/023992、WO2018041122等。其中，目前仅拜耳公司的反义寡核苷酸BAY-2306001进入了临床二期研究。FXIa is currently an emerging target for inhibiting thrombosis. Patent applications that disclose compounds with FXIa inhibitory activity include WO9630396, WO9941276, WO2013093484, WO2004002405, WO2013056060, WO2017005725, WO2017/023992, WO2018041122, etc. Among them, currently only Bayer's antisense oligonucleotide BAY-2306001 has entered the phase II clinical study.
本发明化合物具有更高的活性。特别是本发明化合物表现出优异的对人血液的抗凝血作用，并具有良好的药代活性，可用于有效治疗和/或预防心脑血管疾病及血栓症状。The compounds of the present invention have higher activity. In particular, the compound of the present invention exhibits excellent anticoagulant effects on human blood, and has good pharmacokinetic activity, and can be used to effectively treat and/or prevent cardiovascular and cerebrovascular diseases and thrombotic symptoms.
发明内容Summary of the invention
本发明提供了一系列的氧代哒嗪酰胺类衍生物、其制备方法及其在医药上的应用。The invention provides a series of oxopyridazinamide derivatives, their preparation methods and their medical applications.
具体而言，本发明提供式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其中所有变量如本文所定义。Specifically, the present invention provides a compound of formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein all variables are as defined herein.
Figure PCTCN2020130361-appb-000001
Figure PCTCN2020130361-appb-000001
这些化合物是选择性因子XIa(Factor XIa，简称FXIa)的抑制剂。本发明还涉及包含这些化合物的药物组合物以及使用该化合物治疗血栓栓塞等疾病的药物中的用途。These compounds are inhibitors of selective factor XIa (Factor XIa, FXIa for short). The present invention also relates to a pharmaceutical composition containing these compounds and the use of the compounds in the treatment of diseases such as thromboembolism.
具体的，本发明通过以下技术方案来实现：Specifically, the present invention is implemented through the following technical solutions:
式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐：The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts:
Figure PCTCN2020130361-appb-000002
Figure PCTCN2020130361-appb-000002
R 1选自R 3取代或者未取代的四氮唑，R 3取代或者未取代的三氮唑； R 1 is selected from R 3 substituted or unsubstituted tetrazole, R 3 substituted or unsubstituted triazole;
R 2选自R 4取代或者未取代的苯环，其中R 4选自-NR 5-(CH 2)n-CO-(CH 2)n-NR 6R 7、-NR 5-SO 2-NR 6R 7； R 2 is selected from R 4 substituted or unsubstituted benzene ring, wherein R 4 is selected from -NR 5 -(CH 2 )n-CO-(CH 2 )n-NR 6 R 7 , -NR 5 -SO 2 -NR 6 R 7 ;
Ar选自至少一个R 8取代或者未取代的以下基团：
Figure PCTCN2020130361-appb-000003
Figure PCTCN2020130361-appb-000004
Ar is selected from at least one of the following groups substituted or unsubstituted by R 8:
Figure PCTCN2020130361-appb-000003
Figure PCTCN2020130361-appb-000004
其中，R 3选自氢、卤素、C 1-4的烷基、卤素取代的C 1-4的烷基； Wherein, R 3 is selected from hydrogen, halogen, C 1-4 alkyl, halogen substituted C 1-4 alkyl;
R 5、R 6、R 7独立的选自氢、C 1-4的烷基、C 1-4的烷氧基-C 1-4烷基、-SO 2-C 1-4烷、-SO 2-苯、-C 1-6的一元或者二元醇、
Figure PCTCN2020130361-appb-000005
-(CH 2)n-C 3-12的脂肪环，或者其中NR 5与NR 6R 7之中的任意一个以上通过-(CH 2)n-成环；或者NR 6R 7一起构成C 3-12的脂肪环；前述C 3-12的脂肪环环上的一个以上碳原子被0-2个N、O、S原子所替代，所述脂肪环进一步被一个以上的R 9所取代； R 5 , R 6 , and R 7 are independently selected from hydrogen, C 1-4 alkyl, C 1-4 alkoxy -C 1-4 alkyl, -SO 2 -C 1-4 alkane, -SO 2 -Benzene, -C 1-6 mono or dihydric alcohol,
Figure PCTCN2020130361-appb-000005
-(CH 2 )nC 3-12 aliphatic ring, or wherein any one or more of NR 5 and NR 6 R 7 form a ring through -(CH 2 )n-; or NR 6 R 7 together form C 3-12 The aliphatic ring; one or more carbon atoms on the aforementioned C 3-12 alicyclic ring are replaced by 0-2 N, O, S atoms, and the aliphatic ring is further replaced by more than one R 9 ;
R 8选自氢、卤素、C 1-4的烷基、羟基、-C 1-4的羧酸、-C 1-4的羧酸-C 1-4醇酯； R 8 is selected from hydrogen, halogen, C 1-4 alkyl, hydroxyl, -C 1-4 carboxylic acid, -C 1-4 carboxylic acid-C 1-4 alcohol ester;
R 9选自氢、-(CH 2)n-OH、-SO 2-C 1-4烷、-(CH 2)n-COOH、-酰胺、氰基、NR 10R 11-C 1-4的烷氧基、C 1-4的烷基、C 1-4的烷氧基、C 1-4的烷氧-C 1-4烷基、-CO-吗啉、-CO-NR 12-(CH 2)n-OH、HOOC-C 1-4烷氧基、
Figure PCTCN2020130361-appb-000006
R 9 is selected from hydrogen, -(CH 2 )n-OH, -SO 2 -C 1-4 alkane, -(CH 2 )n-COOH, -amide, cyano, NR 10 R 11 -C 1-4 Alkoxy, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 alkoxy-C 1-4 alkyl, -CO-morpholine, -CO-NR 12 -(CH 2 ) n-OH, HOOC-C 1-4 alkoxy,
Figure PCTCN2020130361-appb-000006
R 10、R 11、R 12独立选自氢或C 1-4烷基； R 10 , R 11 , and R 12 are independently selected from hydrogen or C 1-4 alkyl;
前述n＝0-6的自然数。The aforementioned natural number of n=0-6.
作为本发明的一种优选技术方案，所述-NR 5-(CH 2)n-CO-(CH 2)n-NR 6R 7包括
Figure PCTCN2020130361-appb-000007
Figure PCTCN2020130361-appb-000008
Figure PCTCN2020130361-appb-000009
其中，R 7和R 9如上定义。 As a preferred technical solution of the present invention, the -NR 5 -(CH 2 )n-CO-(CH 2 )n-NR 6 R 7 includes
Figure PCTCN2020130361-appb-000007
Figure PCTCN2020130361-appb-000008
Figure PCTCN2020130361-appb-000009
Wherein, R 7 and R 9 are as defined above.
作为本发明的一种优选技术方案，所述C 1-4的烷基选自甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基； As a preferred technical solution of the present invention, the C 1-4 alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl;
-C 1-4的羧酸选自甲酸、乙酸、丙酸、正丁酸、异丁酸、叔丁酸； -C 1-4 carboxylic acid is selected from formic acid, acetic acid, propionic acid, n-butyric acid, isobutyric acid, tert-butyric acid;
-C 1-4的羧酸-C 1-4醇酯选自甲酸甲酯、甲酸乙酯、甲酸丙酯、甲酸丁酯、乙酸甲酯、乙酸乙酯、乙酸丙酯、乙酸丁酯、丙酸甲酯、丙酸乙酯、丙酸丙酯、丙酸丁酯、丁酸甲酯、丁酸乙酯、丁酸丙酯、丁酸丁酯。 -C 1-4 carboxylic acid-C 1-4 alcohol ester is selected from methyl formate, ethyl formate, propyl formate, butyl formate, methyl acetate, ethyl acetate, propyl acetate, butyl acetate, propyl Methyl acid, ethyl propionate, propyl propionate, butyl propionate, methyl butyrate, ethyl butyrate, propyl butyrate, butyl butyrate.
作为本发明的一种优选技术方案，所述卤素选自氟、氯、溴、碘。As a preferred technical solution of the present invention, the halogen is selected from fluorine, chlorine, bromine, and iodine.
作为本发明的一种优选技术方案，所述C 1-4的烷选自甲烷、乙烷、丙烷、异丙烷、正丁烷、异丁烷、仲丁烷、叔丁烷； As a preferred technical solution of the present invention, the C 1-4 alkane is selected from methane, ethane, propane, isopropane, n-butane, isobutane, sec-butane, and tert-butane;
所述C 1-4的烷氧基选自甲氧基、乙氧基、丙氧基、异丙氧基、正丁氧基、异丁氧基、仲丁氧基、叔丁氧基； The C 1-4 alkoxy group is selected from methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy;
所述-(CH 2)n-OH选自羟基、甲醇、乙醇、正丙醇、正丁醇； The -(CH 2 )n-OH is selected from hydroxyl, methanol, ethanol, n-propanol, and n-butanol;
所述-C 1-6的一元或者二元醇选自甲醇、乙醇、正丙醇、正丁醇、叔丁醇、1,3-丁二醇、3-甲基丁-1-醇、3-甲基戊-1-醇、4-甲基戊-1醇、3-甲基己-1醇、4-甲基己-1醇； The -C 1-6 monohydric or dihydric alcohol is selected from methanol, ethanol, n-propanol, n-butanol, tert-butanol, 1,3-butanediol, 3-methylbutan-1-ol, 3 -Methylpentan-1-ol, 4-methylpentan-1ol, 3-methylhexan-1ol, 4-methylhexan-1ol;
所述NR 10R 11-C 1-4的烷氧基选自甲胺甲氧基、乙胺甲氧基、丙胺甲氧基、丁胺甲氧基、甲胺乙氧基、乙胺乙氧基、丙胺乙氧基、丁胺乙氧基、甲胺丙氧基、乙胺丙氧基、丙胺丙氧基、丁胺丙氧基、甲胺丁氧基、乙胺丁氧基、丙胺丁氧基、丁胺丁氧基、二甲胺基甲氧基、二甲胺基乙氧基、二甲胺基丙氧基、二甲胺基丁氧基； The alkoxy group of the NR 10 R 11 -C 1-4 is selected from the group consisting of methylamine methoxy, ethylamine methoxy, propylamine methoxy, butylamine methoxy, methylamine ethoxy, and ethylamine ethoxy. Propyl, propylamine ethoxy, butylamine ethoxy, methylamine propoxy, ethylamine propoxy, propylamine propoxy, butylamine propoxy, methylamine butoxy, ethylamine butoxy, propylamine Oxy, butylaminobutoxy, dimethylaminomethoxy, dimethylaminoethoxy, dimethylaminopropoxy, dimethylaminobutoxy;
所述C 1-4的烷氧-C 1-4烷基选自甲氧甲基、乙氧甲基、丙氧甲基、丁氧甲基、甲氧乙基、乙氧乙基、丙氧乙基、丁氧乙基、甲氧丙基、乙氧丙基、丙氧丙基、丁氧丙基、甲氧丁基、乙氧丁基、丙氧丁基、丁氧丁基； The C 1-4 alkoxy-C 1-4 alkyl group is selected from methoxymethyl, ethoxymethyl, propoxymethyl, butoxymethyl, methoxyethyl, ethoxyethyl, propoxy Ethyl, butoxyethyl, methoxypropyl, ethoxypropyl, propoxypropyl, butoxypropyl, methoxybutyl, ethoxybutyl, propoxybutyl, butoxybutyl;
所述-CO-NR 12-(CH 2)n-OH选自-CO-NH-CH 2OH、-CO-N(CH 3)-CH 2OH、-CO-NH-CH 2CH 2OH、-CO-N(CH 3)-CH 2CH 2OH； The -CO-NR 12 -(CH 2 )n-OH is selected from -CO-NH-CH 2 OH, -CO-N(CH 3 )-CH 2 OH, -CO-NH-CH 2 CH 2 OH, -CO-N(CH 3 )-CH 2 CH 2 OH;
所述HOOC-C 1-4烷氧基选自HOOC-甲氧基、HOOC-乙氧基、HOOC-丙氧基、HOOC-异丙氧基、HOOC-正丁氧基、HOOC-异丁氧基、HOOC-仲丁氧基、HOOC-叔丁氧基。 The HOOC-C 1-4 alkoxy group is selected from HOOC-methoxy, HOOC-ethoxy, HOOC-propoxy, HOOC-isopropoxy, HOOC-n-butoxy, HOOC-isobutoxy Group, HOOC-sec-butoxy, HOOC-tert-butoxy.
作为本发明的一种优选技术方案，所述C 3-12的脂肪环选自环丙烷、环丁烷、环戊烷、环己烷、环庚烷、环辛烷、环壬烷、环癸烷、螺[3,3]庚环、螺[3,5]壬环、螺[4,5]癸环； As a preferred technical solution of the present invention, the C 3-12 aliphatic ring is selected from cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, and cyclodecane. Alkane, spiro[3,3]heptane ring, spiro[3,5]nonane ring, spiro[4,5]decane ring;
所述C 3-12的脂肪的一个以上碳原子被0-2个N、O、S原子所替代，选自：
Figure PCTCN2020130361-appb-000010
Figure PCTCN2020130361-appb-000011
More than one carbon atom of the C 3-12 fat is replaced by 0-2 N, O, S atoms, selected from:
Figure PCTCN2020130361-appb-000010
Figure PCTCN2020130361-appb-000011
作为本发明的一种优选技术方案，n＝0、1、2、3、4、5、6。As a preferred technical solution of the present invention, n=0,1,2,3,4,5,6.
作为本发明的一种优选技术方案，其特征在于，As a preferred technical solution of the present invention, it is characterized in that:
R 1选自四氮唑，三氮唑； R 1 is selected from tetrazolium and triazole;
R 2选自苯环、
Figure PCTCN2020130361-appb-000012
Figure PCTCN2020130361-appb-000013
Figure PCTCN2020130361-appb-000014
R 2 is selected from benzene ring,
Figure PCTCN2020130361-appb-000012
Figure PCTCN2020130361-appb-000013
Figure PCTCN2020130361-appb-000014
Ar选自以下基团：
Figure PCTCN2020130361-appb-000015
Figure PCTCN2020130361-appb-000016
Ar is selected from the following groups:
Figure PCTCN2020130361-appb-000015
Figure PCTCN2020130361-appb-000016
作为本发明的一种优选方案，所述化合物或其药学上可接受的盐，选自以下化合物：As a preferred embodiment of the present invention, the compound or a pharmaceutically acceptable salt thereof is selected from the following compounds:
Figure PCTCN2020130361-appb-000017
Figure PCTCN2020130361-appb-000017
Figure PCTCN2020130361-appb-000018
Figure PCTCN2020130361-appb-000018
Figure PCTCN2020130361-appb-000019
Figure PCTCN2020130361-appb-000019
Figure PCTCN2020130361-appb-000020
Figure PCTCN2020130361-appb-000020
Figure PCTCN2020130361-appb-000021
Figure PCTCN2020130361-appb-000021
Figure PCTCN2020130361-appb-000022
Figure PCTCN2020130361-appb-000022
Figure PCTCN2020130361-appb-000023
Figure PCTCN2020130361-appb-000023
Figure PCTCN2020130361-appb-000024
Figure PCTCN2020130361-appb-000024
Figure PCTCN2020130361-appb-000025
Figure PCTCN2020130361-appb-000025
作为本发明的一种优选方案，所述药学上可接受的盐是指化合物与药学上可接受的酸或碱制备。As a preferred embodiment of the present invention, the pharmaceutically acceptable salt refers to a compound prepared with a pharmaceutically acceptable acid or base.
作为本发明的一种优选方案，所述化合物的一个以上的氢原子上被同位素氘取代。As a preferred solution of the present invention, more than one hydrogen atom of the compound is replaced by an isotope deuterium.
本发明另一目的提供了一种药物组合物，包括前述的式(I)的化合物，或其立体异构体、互变异构体、药学上可接受的盐和一种以上药学上可接受的载体。Another object of the present invention is to provide a pharmaceutical composition comprising the aforementioned compound of formula (I), or its stereoisomers, tautomers, pharmaceutically acceptable salts and more than one pharmaceutically acceptable a.
本发明另一目的在于提供所述的式(I)的化合物，或其立体异构体、互变异构体、药学上可接受的盐、以及含有所述化合物，或其立体异构体、互变异构体、药学上可接受的盐的药物组合物在制备用于制备治疗FXIa相关疾病的药物用途，具体地，涉及血栓相关疾病的药物用途。Another object of the present invention is to provide the compound of formula (I), or its stereoisomers, tautomers, pharmaceutically acceptable salts, and containing the compound, or its stereoisomers, The pharmaceutical composition of tautomers and pharmaceutically acceptable salts is used in the preparation of pharmaceuticals for the treatment of FXIa-related diseases, in particular, the pharmaceutical uses related to thrombosis-related diseases.
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A specific term or phrase should not be considered uncertain or unclear without a special definition, but should be understood in its ordinary meaning. When a trade name appears in this article, it is meant to refer to its corresponding commodity or its active ingredient. The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and/or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues. , Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与药学上可接受的酸或碱制备。The term "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention, which is prepared from the compound with specific substituents discovered in the present invention and a pharmaceutically acceptable acid or base.
所述
Figure PCTCN2020130361-appb-000026
为连接键。 Said
Figure PCTCN2020130361-appb-000026
It is the connection key.
除了盐的形式，本发明所提供的化合物还存在前药形式。本文所描述的化合物的前药容易地在生理条件下发生化学变化从而转化成本发明的化合物。此外，前体药物可以在体内环境中通过化学或生化方法被转换到本发明的化合物。In addition to salt forms, the compounds provided by the present invention also exist in prodrug forms. The prodrugs of the compounds described herein easily undergo chemical changes under physiological conditions to transform into the compounds of the invention. In addition, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in the in vivo environment.
本发明的某些化合物可以以非溶剂化形式或者溶剂化形式存在，包括水合物形式。一般而言，溶剂化形式与非溶剂化的形式相当，都包含在本发明的范围之内。Certain compounds of the present invention may exist in unsolvated or solvated forms, including hydrated forms. Generally speaking, the solvated form is equivalent to the unsolvated form, and both are included in the scope of the present invention.
本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers Conformers, (D)-isomers, (L)-isomers, and their racemic mixtures and other mixtures, such as enantiomers or diastereomer-enriched mixtures, all of these mixtures belong to Within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All these isomers and their mixtures are included in the scope of the present invention.
可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体，以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备，其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。The optically active (R)- and (S)-isomers, as well as the D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If you want to obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with chiral auxiliary agents, in which the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure The desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), it forms a diastereomeric salt with an appropriate optically active acid or base, and then passes through a conventional method known in the art The diastereoisomers are resolved, and then the pure enantiomers are recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished through the use of chromatography, which uses a chiral stationary phase and is optionally combined with chemical derivatization (for example, the formation of amino groups from amines). Formate).
本发明化合物分子的原子是同位素，通过同位素衍生化通常可以延长半衰期、降低清除率、增强代谢稳定和提高体内活性等效果。并且，包括一个实施方案，其中至少一个原子被具有相同原子数(质子数)和不同质量数(质子和中子和)的原子取代。本发明化合物中包括的同位素的实例包括氢原子、碳原子、氮原子、氧原子、磷原子、硫原子、氟原子、氯原子，其分别包括 2H、 3H、 13C、 14C、 15N、 17O、 18O、 31P、 32P、 35S、 18F、 36Cl。特别的是，随其衰退而发射辐射的放射性同位素例如 3H或 14C可用于药物制剂或者体内化合物的局部解剖学检验。稳定的同位素既不随其量衰减或变化，也不具有放射性，因此其可以安全使用。当构成本发明化合物分子的原子是同位素时，通过用包含相应同位素的试剂替代合成中所用的试剂，可以根据通用方法转化同位素。 The atoms of the compound molecules of the present invention are isotopes, and isotope derivatization can generally extend the half-life, reduce the clearance rate, enhance the stability of metabolism, and increase the activity in the body. And, an embodiment is included in which at least one atom is substituted with atoms having the same atomic number (number of protons) and different mass numbers (sum of protons and neutrons). Examples of isotopes included in the compounds of the present invention include hydrogen atoms, carbon atoms, nitrogen atoms, oxygen atoms, phosphorus atoms, sulfur atoms, fluorine atoms, and chlorine atoms, which respectively include 2 H, 3 H, 13 C, 14 C, and 15 N, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 Cl. In particular, radioisotopes that emit radiation as they decay, such as 3 H or 14 C, can be used for topographical examinations of pharmaceutical preparations or compounds in the body. The stable isotope neither decays or changes with its amount, nor is it radioactive, so it can be used safely. When the atoms constituting the molecule of the compound of the present invention are isotopes, the isotopes can be converted according to general methods by replacing the reagents used in the synthesis with reagents containing the corresponding isotopes.
本发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氘( 2H)，碘-125( 125I)或C-14( 14C)。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。 The compound of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms constituting the compound. For example, compounds can be labeled with radioisotopes, such as deuterium ( 2 H), iodine-125 ( 125 I), or C-14 ( 14 C). All changes in the isotopic composition of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.
进一步地，本发明的化合物一个或多个氢原子上被同位素氘( 2H)取代，本发明化合物氘代后，具有延长半衰期、降低清除率、增强代谢稳定和提高体内活性等效果。 Further, one or more hydrogen atoms of the compound of the present invention are replaced by the isotope deuterium ( 2 H). After deuteration, the compound of the present invention has the effects of prolonging half-life, reducing clearance, enhancing metabolic stability, and improving in vivo activity.
所述同位素衍生物的制备方法通常包括：相转移催化方法。例如，优选的氘化方法采用相转移催化剂(例如，四烷基铵盐，NBu 4HSO 4)。使用相转移催化剂交换二苯基甲烷化合物的亚甲基质子，导致比在酸(例如，甲磺酸)存在下用氘化硅烷(例如三乙基氘化甲硅烷)或用路易斯酸如三氯化铝采用氘化硼酸钠还原而引入较高的氘。 The preparation method of the isotope derivative usually includes a phase transfer catalysis method. For example, the preferred deuteration method uses a phase transfer catalyst (e.g., tetraalkylammonium salt, NBu 4 HSO 4 ). The use of a phase transfer catalyst to exchange the methylene protons of the diphenylmethane compound results in the use of deuterated silanes (e.g. triethyl deuterated monosilane) or Lewis acids such as trichlorosilane in the presence of an acid (e.g., methanesulfonic acid) Aluminum chloride is reduced with deuterated sodium borate to introduce higher deuterium.
术语“药学上可接受的载体”是指能够递送本发明有效量活性物质、不干扰活性物质的生物活性并且对宿主或者患者无毒副作用的任何制剂载体或介质，代表性的载体包括水、油、蔬菜和矿物质、膏基、洗剂基质、软膏基质等。这些基质包括悬浮剂、增粘剂、透皮促进剂等。它们的制剂为化妆品领域或局部药物领域的技术人员所周知。关于载体的其他信息，可以参考Remington:The Science and Practice of Pharmacy,21st Ed.,Lippincott,Williams&Wilkins(2005)，该文献的内容通过引用的方式并入本文。The term "pharmaceutically acceptable carrier" refers to any preparation carrier or medium that can deliver an effective amount of the active substance of the present invention, does not interfere with the biological activity of the active substance, and has no toxic or side effects to the host or patient. Representative carriers include water and oil. , Vegetables and minerals, cream base, lotion base, ointment base, etc. These bases include suspending agents, tackifiers, penetration enhancers and the like. Their formulations are well known to those skilled in the field of cosmetics or topical medicine. For other information about the carrier, you can refer to Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), and the content of this document is incorporated herein by reference.
术语“赋形剂”通常是指配制有效的药物组合物所需要载体、稀释剂和/或介质。The term "excipient" generally refers to the carrier, diluent and/or medium required to formulate an effective pharmaceutical composition.
针对药物或药理学活性剂而言，术语“有效量”或“治疗有效量”是指无毒的但能达到预期效果的药物或药剂的足够用量。对于本发明中的口服剂型，组合物中一种活性物质的“有效量”是指与该组合物中另一种活性物质联用时为了达到预期效果所需要的用量。有效量的确定因人而异，取决于受体的年龄和一般情况，也取决于具体的活性物质，个案中合适的有效量可以由本领域技术人员根据常规试验确定。For drugs or pharmacologically active agents, the term "effective amount" or "therapeutically effective amount" refers to a sufficient amount of a drug or agent that is non-toxic but can achieve the desired effect. For the oral dosage form of the present invention, the "effective amount" of one active substance in the composition refers to the amount required to achieve the desired effect when combined with another active substance in the composition. The determination of the effective amount varies from person to person, and depends on the age and general conditions of the recipient, as well as the specific active substance. The appropriate effective amount in a case can be determined by those skilled in the art according to routine experiments.
术语“活性成分”、“治疗剂”，“活性物质”或“活性剂”是指一种化学实体，它可以有效地治疗目标紊乱、疾病或病症。The terms "active ingredient", "therapeutic agent", "active substance" or "active agent" refer to a chemical entity that can effectively treat the target disorder, disease or condition.
“任选”或“任选地”指的是随后描述的事件或状况可能但不是必需出现的，并且该描述包括其中所述事件或状况发生的情况以及所述事件或状况不发生的情况。"Optional" or "optionally" means that the event or condition described later may but not necessarily occur, and the description includes a situation in which the event or condition occurs and a situation in which the event or condition does not occur.
本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and those well known to those skilled in the art Equivalent alternatives, preferred implementations include but are not limited to the embodiments of the present invention.
具体实施方式Detailed ways
下面结合实施例对本发明作进一步详细的描述，但发明的实施方式不限于此。The present invention will be further described in detail below in conjunction with examples, but the implementation of the invention is not limited thereto.
化合物的结构是通过核磁共振(NMR)或质谱(MS)来确定的。NMR位移(δ)以10-6(ppm)的单位给出。NMR的测定是用Bruker AVANCE-III核磁仪，测定溶剂为氘代二甲基亚砜(DMSO-d6)，氘代氯仿(CDCl3)，内标为四甲基硅烷(TMS)。The structure of the compound is determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). The NMR shift (δ) is given in units of 10-6 (ppm). NMR was measured with Bruker AVANCE-III nuclear magnetometer, the solvent was deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and the internal standard was tetramethylsilane (TMS).
MS的测定用ISQ EC质谱仪(生产商:Thermo,型号:ISQ EC)。MS is measured with ISQ EC mass spectrometer (manufacturer: Thermo, model: ISQ EC).
高效液相色谱法(HPLC)分析使用Thermo U3000 HPLC DAD高效液相色谱仪。High performance liquid chromatography (HPLC) analysis uses Thermo U3000 HPLC DAD high performance liquid chromatograph.
CombiFlash快速制备仪使用CombiFlash Rf+LUMEN(TELEDYNE ISCO)。CombiFlash rapid preparation instrument uses CombiFlash Rf+LUMEN (TELEDYNE ISCO).
薄层层析硅胶板使用烟台银龙HSGF254或GF254硅胶板，薄层色谱法(TLC)使用的硅胶板采用的规格是0.17mm～0.23mm，薄层层析分离纯化产品采用的规格是0.4mm～0.5mm。The thin layer chromatography silica gel plate uses Yantai Yinlong HSGF254 or GF254 silica gel plate. The size of the silica gel plate used in thin layer chromatography (TLC) is 0.17mm～0.23mm, and the size of thin layer chromatography separation and purification products is 0.4mm. ~0.5mm.
硅胶柱色谱法一般使用乳山上邦硅胶100～200目硅胶为载体。The silica gel column chromatography generally uses Rushan Shangbang silica gel 100-200 mesh silica gel as the carrier.
实施例1Example 1
合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(2-氧代-2,3-二氢-1H-苯并[d]咪唑-5-基)-3-苯基丙酰胺Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N-(2 -Oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-3-phenylpropionamide
Figure PCTCN2020130361-appb-000027
Figure PCTCN2020130361-appb-000027
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成2-(5-氯-2-硝基苯基)氨基)-2-氧代乙酸Step A: Synthesis of 2-(5-chloro-2-nitrophenyl)amino)-2-oxoacetic acid
Figure PCTCN2020130361-appb-000028
Figure PCTCN2020130361-appb-000028
将5-氯-2-硝基苯胺(4.0克，23.2毫摩尔)溶于二氯甲烷(30.0毫升)，加入草酰氯(7.3克，58.2毫摩尔)室温反应一个小时随后在冰水浴下缓慢加水，有气泡生成并有淡黄色固体析出，加水至不再有气泡生成，停止反应。Dissolve 5-chloro-2-nitroaniline (4.0 g, 23.2 mmol) in dichloromethane (30.0 mL), add oxalyl chloride (7.3 g, 58.2 mmol) and react at room temperature for one hour, then slowly add water in an ice-water bath , Bubbles are formed and light yellow solids are precipitated. Add water until no more bubbles are formed to stop the reaction.
反应液在真空条件下旋走二氯甲烷，过滤，得浅黄色滤饼，滤饼于真空干燥箱烘干得5.2克浅黄色固体2-(5-氯-2-硝基苯基)氨基)-2-氧代乙酸粗品(收率：91.7％)。MS(ESI)M/Z：[M+H] +＝244.98。 The reaction solution was spun off the dichloromethane under vacuum and filtered to obtain a light yellow filter cake. The filter cake was dried in a vacuum drying oven to obtain 5.2 g of light yellow solid 2-(5-chloro-2-nitrophenyl)amino) Crude -2-oxoacetic acid (yield: 91.7%). MS (ESI) M/Z: [M+H] + =244.98.
步骤B：合成(2-((5-氯-2-硝基苯基)氨基)-2-氧代乙酰基)-L-苯丙氨酸叔丁酯Step B: Synthesis of (2-((5-chloro-2-nitrophenyl)amino)-2-oxoacetyl)-L-phenylalanine tert-butyl ester
Figure PCTCN2020130361-appb-000029
Figure PCTCN2020130361-appb-000029
将2-(5-氯-2-硝基苯基)氨基)-2-氧代乙酸(5.20克，21.31毫摩尔)和L-苯丙氨酸叔丁酯盐酸盐(6.57克，25.57毫摩尔)溶于二氯甲烷(30.0毫升)中，加入N，N-二异丙基乙胺(8.25克，63.93毫摩尔)，加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(12.15克，31.96毫摩尔)。随后，在室温下搅拌2小时。Combine 2-(5-chloro-2-nitrophenyl)amino)-2-oxoacetic acid (5.20 g, 21.31 mmol) and L-phenylalanine tert-butyl ester hydrochloride (6.57 g, 25.57 mmol) Mol) was dissolved in dichloromethane (30.0 ml), N,N-diisopropylethylamine (8.25 g, 63.93 mmol) was added, and 2-(7-benzotriazole oxide)-N,N ,N',N'-Tetramethylurea hexafluorophosphate (12.15g, 31.96mmol). Subsequently, it was stirred at room temperature for 2 hours.
向反应液中加入水(20毫升)淬灭反应。加入乙酸乙酯(150毫升)，混合液用水和饱和食盐水(30毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/10)，得到5.80克淡黄色固体(2-((5-氯-2-硝基苯基)氨基)-2-氧代乙酰基)-L-苯丙氨酸叔丁酯(收率：61.0％)。MS(ESI)M/Z：[M+H] +＝448.12。 Water (20 mL) was added to the reaction solution to quench the reaction. Ethyl acetate (150 mL) was added, and the mixture was washed with water and saturated brine (30 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/10) to obtain 5.80 g of pale yellow solid (2-((5-chloro-2-nitrophenyl)amino) -2-oxoacetyl)-L-phenylalanine tert-butyl ester (yield: 61.0%). MS (ESI) M/Z: [M+H] + =448.12.
步骤C：合成(S)-2-(4-(5-氯-2-硝基苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸叔丁酯Step C: Synthesis of (S)-2-(4-(5-chloro-2-nitrophenyl)-2,3-dioxopiperazin-1-yl)-3-phenylpropionic acid tert-butyl ester
Figure PCTCN2020130361-appb-000030
Figure PCTCN2020130361-appb-000030
将(2-((5-氯-2-硝基苯基)氨基)-2-氧代乙酰基)-L-苯丙氨酸叔丁酯(5.8克，12.97毫摩尔)溶于无水乙腈(30.0毫升)，加入碳酸钾(8.9克，64.85毫摩尔)，1,2-二溴乙烷(9.6克，51.90毫摩尔)，氮气置换三次，升温至100摄氏度反应16小时。Dissolve (2-((5-chloro-2-nitrophenyl)amino)-2-oxoacetyl)-L-phenylalanine tert-butyl ester (5.8 g, 12.97 mmol) in anhydrous acetonitrile (30.0 ml), potassium carbonate (8.9 g, 64.85 mmol), 1,2-dibromoethane (9.6 g, 51.90 mmol) were added, replaced with nitrogen three times, and the temperature was raised to 100 degrees Celsius to react for 16 hours.
向反应液中加入乙酸乙酯(300毫升)，混合液用水和饱和食盐水(50毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/1)，得到3.0克黄色固体(S)-2-(4-(5-氯-2-硝基苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸叔丁酯(收率：49.0％)。MS(ESI)M/Z：[M+H] +＝474.13。 Ethyl acetate (300 mL) was added to the reaction solution, and the mixture was washed with water and saturated brine (50 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/1) to obtain 3.0 g of yellow solid (S)-2-(4-(5-chloro-2-nitrobenzene) (Yl)-2,3-dioxopiperazin-1-yl)-3-phenylpropionic acid tert-butyl ester (yield: 49.0%). MS (ESI) M/Z: [M+H] + =474.13.
步骤D：合成(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸叔丁酯Step D: Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-phenylpropionic acid tert-butyl ester
Figure PCTCN2020130361-appb-000031
Figure PCTCN2020130361-appb-000031
将(S)-2-(4-(5-氯-2-硝基苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸叔丁酯(3.0克，6.34毫摩尔)溶于乙酸乙酯溶液(35毫升)中，加入干钯碳(300毫克，10％摩尔)。置换氢气三次，于室温下反应5小时。Add (S)-2-(4-(5-chloro-2-nitrophenyl)-2,3-dioxopiperazin-1-yl)-3-phenylpropionic acid tert-butyl ester (3.0 g , 6.34 mmol) was dissolved in ethyl acetate solution (35 mL), and dry palladium on carbon (300 mg, 10% mol) was added. The hydrogen was replaced three times and reacted at room temperature for 5 hours.
反应结束，过滤掉钯碳，滤饼层用甲醇(10毫升×3次)洗涤，滤液减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/1)，得到1.0克淡黄色固体(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸叔丁酯(收率：35.7％)。MS(ESI)M/Z：[M+H] +＝444.16。 After the reaction was completed, the palladium carbon was filtered off, the filter cake layer was washed with methanol (10 ml×3 times), the filtrate was concentrated under reduced pressure, and the residue obtained was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/ 1) to obtain 1.0 g of light yellow solid (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-phenylpropane Tert-butyl ester (yield: 35.7%). MS(ESI) M/Z: [M+H] + =444.16.
步骤E：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸叔丁酯Step E: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Tert-butyl phenylpropionate
Figure PCTCN2020130361-appb-000032
Figure PCTCN2020130361-appb-000032
将(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸叔丁酯(1.0克，2.26毫摩尔)溶于乙酸(15.0毫升)，于冰水浴下，加入原甲酸三乙酯(1.0克，6.77毫摩尔)，叠氮化钠(440毫克，6.77毫摩尔)，缓慢升至80摄氏度继续反应16小时。(S)-2-(4-(2-Amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-phenylpropionic acid tert-butyl ester (1.0 g, 2.26 mmol) was dissolved in acetic acid (15.0 ml), under ice-water bath, added triethyl orthoformate (1.0 g, 6.77 mmol), sodium azide (440 mg, 6.77 mmol), and slowly increased to 80 degrees Celsius Continue the reaction for 16 hours.
向反应液中加水，有类白色沉淀析出，继续加水至无固体洗出，过滤，滤饼用水(10毫升×3次)洗涤，干燥得523毫克类白色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸叔丁酯(收率：46.0％)。MS(ESI)M/Z：[M+H] +＝497.16。 Water was added to the reaction solution, an off-white precipitate precipitated. Continue to add water until no solid was washed out, filtered, and the filter cake was washed with water (10 ml×3 times) and dried to obtain 523 mg of off-white solid (S)-2-(4- (5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenylpropionic acid tert-butyl ester (Yield: 46.0 %). MS(ESI) M/Z: [M+H] + =497.16.
步骤F：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸Step F: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionic acid
Figure PCTCN2020130361-appb-000033
Figure PCTCN2020130361-appb-000033
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸叔丁酯(523毫克，2.26毫摩尔)溶于二氯甲烷(10.0毫升)，加入三氟乙酸(2.0毫升)，于室温反应3小时。(S)-2-(4-(5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenyl Tert-butyl propionate (523 mg, 2.26 mmol) was dissolved in dichloromethane (10.0 mL), trifluoroacetic acid (2.0 mL) was added, and the reaction was carried out at room temperature for 3 hours.
于低温下旋走二氯甲烷和三氟乙酸，得450毫克淡黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(收率：97.0％)。MS(ESI)M/Z：[M+H] +＝441.10。 Rotate off dichloromethane and trifluoroacetic acid at low temperature to obtain 450 mg of light yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2 ,3-dioxopiperazin-1-yl)-3-phenylpropionic acid (yield: 97.0%). MS(ESI) M/Z: [M+H] + =441.10.
步骤G：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(2-氧代-2,3-二氢-1H-苯并[d]咪唑-5-基)-3-苯基丙酰胺Step G: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N -(2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-3-phenylpropionamide
Figure PCTCN2020130361-appb-000034
Figure PCTCN2020130361-appb-000034
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(46毫克，0.10毫摩尔)，5-氨基-1,3-二氢-2H-苯并[d]咪唑-2-酮(17.8毫克，0.12毫摩尔)溶于N,N-二甲基甲酰胺(2.5毫升)中，加入N,N-二异丙基乙胺(65毫克，0.50毫摩尔)。随后，向上述溶液中加入1-丙基磷酸酐(127毫克，0.40毫摩尔),室温反应18小时。(S)-2-(4-(5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenyl Propionic acid (46 mg, 0.10 mmol), 5-amino-1,3-dihydro-2H-benzo[d]imidazol-2-one (17.8 mg, 0.12 mmol) dissolved in N,N-dimethyl N,N-diisopropylethylamine (65 mg, 0.50 mmol) was added to methyl formamide (2.5 mL). Subsequently, 1-propyl phosphoric anhydride (127 mg, 0.40 mmol) was added to the above solution, and reacted at room temperature for 18 hours.
加水淬灭反应，有固体析出，过滤，滤饼，得粗产品70毫克，粗产品用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，纯化得50毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(2-氧代-2,3-二氢-1H-苯并[d]咪唑-5-基)-3-苯基丙酰胺(收率：54.0％)。MS(ESI)M/Z：[M+H] +＝572.15。 1H NMR(400MHz,DMSO)δ10.57(d,J＝28.5Hz,2H),10.13(s,1H),9.77(s,1H),7.94(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.43(d,J＝1.9Hz,1H),7.37–7.22(m,5H),7.05(dd,J＝8.4,1.9Hz,1H),6.85(t,J＝8.4Hz,1H),5.32(s,1H),4.04(dd,J＝14.2,7.1Hz,2H),3.73(s,2H),3.28(d,J＝5.6Hz,1H),3.08(dd,J＝14.2,10.0Hz,1H)。 The reaction was quenched by adding water, a solid was precipitated, filtered, and a filter cake was used to obtain 70 mg of crude product. The crude product was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, 50 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1 was obtained -Yl)-N-(2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-3-phenylpropionamide (yield: 54.0%). MS (ESI) M/Z: [M+H] + =572.15. 1 H NMR(400MHz,DMSO)δ10.57(d,J=28.5Hz,2H),10.13(s,1H),9.77(s,1H),7.94(d,J=2.2Hz,1H),7.82( d,J=8.6Hz,1H),7.77(dd,J=8.6,2.3Hz,1H),7.43(d,J=1.9Hz,1H),7.37–7.22(m,5H),7.05(dd,J ＝8.4,1.9Hz,1H),6.85(t,J＝8.4Hz,1H),5.32(s,1H),4.04(dd,J＝14.2,7.1Hz,2H),3.73(s,2H),3.28 (d, J = 5.6 Hz, 1H), 3.08 (dd, J = 14.2, 10.0 Hz, 1H).
实施例2Example 2
合成(S)-4-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido) benzoic acid
Figure PCTCN2020130361-appb-000035
Figure PCTCN2020130361-appb-000035
步骤A：合成(S)-4-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020130361-appb-000036
Figure PCTCN2020130361-appb-000036
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(83毫克，0.18毫摩尔)，4-氨基苯甲酸叔丁酯(43.4毫克，0.22毫摩尔)溶于N,N-二甲基甲酰胺(2.5毫升)中，加入N,N-二异丙基乙胺(122毫克，0.94毫摩尔)。随后，向上述溶液中加入1-丙基磷酸酐(239毫克，0.75毫摩尔)。将室温反应18小时。(S)-2-(4-(5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenyl Propionic acid (83 mg, 0.18 mmol), tert-butyl 4-aminobenzoate (43.4 mg, 0.22 mmol) were dissolved in N,N-dimethylformamide (2.5 mL), and N,N-di Isopropylethylamine (122 mg, 0.94 mmol). Subsequently, 1-propyl phosphoric anhydride (239 mg, 0.75 mmol) was added to the above solution. React at room temperature for 18 hours.
将反应液乙酸乙酯(30毫升)稀释，用水和饱和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏。将所得残余物TLC板纯化(乙酸乙酯/正己烷＝1:1)得20毫克淡黄色固体(S)-4-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：17.0％)。MS(ESI)M/Z：[M+H] +＝616.20。 The reaction solution was diluted with ethyl acetate (30 mL), washed with water and saturated brine (10 mL×3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The obtained residue was purified by TLC plate (ethyl acetate/n-hexane=1:1) to obtain 20 mg of pale yellow solid (S)-4-(2-(4-(5-chloro-2-(1H-tetrazole- 1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenylpropionamido)tert-butyl benzoate (yield: 17.0%). MS(ESI) M/Z: [M+H] + =616.20.
步骤B：合成(S)-4-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸Step B: Synthesis of (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)benzoic acid
Figure PCTCN2020130361-appb-000037
Figure PCTCN2020130361-appb-000037
将(S)-4-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸叔丁酯(20.0毫克，0.03毫摩尔)溶于二氯甲烷(3.0毫升)。随后，向上述溶液中加入三氟乙酸(0.6毫升)。室温反应3小时。(S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido) tert-butyl benzoate (20.0 mg, 0.03 mmol) was dissolved in dichloromethane (3.0 mL). Subsequently, trifluoroacetic acid (0.6 mL) was added to the above solution. React at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，纯化得13.7毫克黄色固体(S)-4-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸(收率：75.0％)。MS(ESI)M/Z：[M+H] +＝572.15。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, 13.7 mg of yellow solid (S)-4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper was obtained after purification (Azin-1-yl)-3-phenylpropionamido)benzoic acid (yield: 75.0%). MS (ESI) M/Z: [M+H] + =572.15.
实施例3Example 3
合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(3-氧代异吲哚啉-5-基)-3-苯基丙酰胺Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N-(3 -Oxoisoindolin-5-yl)-3-phenylpropionamide
Figure PCTCN2020130361-appb-000038
Figure PCTCN2020130361-appb-000038
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(3-氧代异吲哚啉-5-基)-3-苯基丙酰胺Step A: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N -(3-oxoisoindolin-5-yl)-3-phenylpropionamide
Figure PCTCN2020130361-appb-000039
Figure PCTCN2020130361-appb-000039
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(47毫克，0.10毫摩尔)，6-氨基异吲哚啉-1-酮(17毫克，0.12毫摩尔)溶于N,N-二甲基甲酰胺(2.0毫升)中，加入N,N-二异丙基乙胺(69毫克，0.53毫摩尔)。 随后，向上述溶液中加入1-丙基磷酸酐(136毫克，0.43毫摩尔),室温反应18小时。(S)-2-(4-(5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenyl Propionic acid (47 mg, 0.10 mmol), 6-aminoisoindolin-1-one (17 mg, 0.12 mmol) were dissolved in N,N-dimethylformamide (2.0 mL), and N, N-Diisopropylethylamine (69 mg, 0.53 mmol). Subsequently, 1-propyl phosphoric anhydride (136 mg, 0.43 mmol) was added to the above solution, and reacted at room temperature for 18 hours.
加水淬灭反应，二氯甲烷(10毫升×3次)萃取，减压蒸馏，所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，纯化得11.98毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(3-氧代异吲哚啉-5-基)-3-苯基丙酰胺(收率：20.0％)。MS(ESI)M/Z：[M+H] +＝571.15。 1H NMR(400MHz,DMSO)δ10.40(s,1H),9.75(s,1H),8.56(s,1H),8.03(d,J＝1.3Hz,1H),7.93(d,J＝2.2Hz,1H),7.80(d,J＝8.6Hz,1H),7.75(dd,J＝8.6,2.2Hz,1H),7.69(dd,J＝8.2,2.0Hz,1H),7.52(d,J＝8.2Hz,1H),7.41–7.15(m,5H),5.33(s,1H),4.37–4.30(m,2H),4.10–3.91(m,2H),3.73(s,2H),3.37(dd,J＝11.0,5.8Hz,1H),3.16–3.05(m,1H)。 The reaction was quenched by adding water, extracted with dichloromethane (10 ml×3 times), and distilled under reduced pressure. The residue obtained was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was purified to obtain 11.98 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1 -Yl)-N-(3-oxoisoindolin-5-yl)-3-phenylpropionamide (yield: 20.0%). MS(ESI) M/Z: [M+H] + =571.15. 1 H NMR (400MHz, DMSO) δ 10.40 (s, 1H), 9.75 (s, 1H), 8.56 (s, 1H), 8.03 (d, J = 1.3 Hz, 1H), 7.93 (d, J = 2.2 Hz, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.75 (dd, J = 8.6, 2.2 Hz, 1H), 7.69 (dd, J = 8.2, 2.0 Hz, 1H), 7.52 (d, J =8.2Hz,1H),7.41–7.15(m,5H),5.33(s,1H), 4.37–4.30(m,2H), 4.10–3.91(m,2H), 3.73(s,2H), 3.37( dd, J=11.0, 5.8 Hz, 1H), 3.16-3.05 (m, 1H).
实施例4Example 4
合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(3-氧代异吲哚啉-5-基)-3-苯基丙酰胺Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N-(3 -Oxoisoindolin-5-yl)-3-phenylpropionamide
Figure PCTCN2020130361-appb-000040
Figure PCTCN2020130361-appb-000040
具体合成路线如下：The specific synthesis route is as follows:
步骤A:合成(S)-5-(3-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-叔丁酯苯基丙氧基)苯基)-3-氧代-2,3-二氢-1H-吡唑-1-羧酸叔丁酯Step A: Synthesis of (S)-5-(3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-tert-butyl phenylpropoxy)phenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000041
Figure PCTCN2020130361-appb-000041
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(88毫克，0.20毫摩尔)，5-(3-氨基苯基)-3-氧代-2,3-二氢-1H-吡唑-1-羧酸叔丁酯(66毫克，0.24毫摩尔)溶于N,N-二甲基甲酰胺(2.0毫升)中，加入N,N-二异丙基乙胺(129毫克，1.0毫摩尔)。随后，向上述溶液中加入1-丙基磷酸酐(254毫克，0.8毫摩尔),室温反应18小时。(S)-2-(4-(5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenyl Propionic acid (88 mg, 0.20 mmol), tert-butyl 5-(3-aminophenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylate (66 mg, 0.24 (Mmol) was dissolved in N,N-dimethylformamide (2.0 mL), and N,N-diisopropylethylamine (129 mg, 1.0 mmol) was added. Subsequently, 1-propyl phosphoric anhydride (254 mg, 0.8 mmol) was added to the above solution, and reacted at room temperature for 18 hours.
加水淬灭反应，加乙酸乙酯(20毫升)稀释，水和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏，所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，纯化得42.8毫克黄色固体(S)-5-(3-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-叔丁酯苯基丙氧基)苯基)-3-氧代-2,3-二氢-1H-吡唑-1-羧酸叔丁酯(收率：30.7％)。MS(ESI)M/Z：[M+H] +＝698.21。 The reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml×3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue obtained was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, 42.8 mg of yellow solid (S)-5-(3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-bis (Oxopiperazin-1-yl)-3-tert-butyl phenylpropoxy)phenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylic acid tert-butyl ester ( Yield: 30.7%). MS (ESI) M/Z: [M+H] + =698.21.
实施例5Example 5
合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(4-(5-氧代-2-5-二氢-1H-吡唑-3-基)苯基)-3-苯基丙酰胺Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N-(4 -(5-oxo-2-5-dihydro-1H-pyrazol-3-yl)phenyl)-3-phenylpropionamide
Figure PCTCN2020130361-appb-000042
Figure PCTCN2020130361-appb-000042
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(4-(5-氧代-2-5-二氢-1H-吡唑-3-基)苯 基)-3-苯基丙酰胺Step A: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-N -(4-(5-oxo-2-5-dihydro-1H-pyrazol-3-yl)phenyl)-3-phenylpropionamide
Figure PCTCN2020130361-appb-000043
Figure PCTCN2020130361-appb-000043
将(S)-5-(3-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-叔丁酯苯基丙氧基)苯基)-3-氧代-2,3-二氢-1H-吡唑-1-羧酸叔丁酯(40.0毫克，0.057毫摩尔)溶于二氯甲烷(3.0毫升)。随后，向上述溶液中加入三氟乙酸(0.6毫升)。室温反应3小时。(S)-5-(3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-tert-butyl phenylpropoxy)phenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylic acid tert-butyl ester (40.0 mg, 0.057 mmol) In dichloromethane (3.0 mL). Subsequently, trifluoroacetic acid (0.6 mL) was added to the above solution. React at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，纯化得3.4毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-N-(4-(5-氧代-2-5-二氢-1H-吡唑-3-基)苯基)-3-苯基丙酰胺(收率：10.0％)。MS(ESI)M/Z：[M+H] +＝598.16。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, 3.4 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1 was obtained -Yl)-N-(4-(5-oxo-2-5-dihydro-1H-pyrazol-3-yl)phenyl)-3-phenylpropionamide (yield: 10.0%). MS(ESI) M/Z: [M+H] + =598.16.
实施例6Example 6
合成(S)-3-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido) benzoic acid
Figure PCTCN2020130361-appb-000044
Figure PCTCN2020130361-appb-000044
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-3-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020130361-appb-000045
Figure PCTCN2020130361-appb-000045
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(43毫克，0.098毫摩尔)，3-氨基苯甲酸叔丁酯(23毫克，0.117毫摩尔)溶于N,N-二甲基甲酰胺(2.0毫升)中，加入N,N-二异丙基乙胺(63毫克，0.49毫摩尔)。随后，向上述溶液中加入1-丙基磷酸酐(125毫克，0.392毫摩尔),室温反应18小时。(S)-2-(4-(5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenyl Propionic acid (43 mg, 0.098 mmol), tert-butyl 3-aminobenzoate (23 mg, 0.117 mmol) were dissolved in N,N-dimethylformamide (2.0 mL), and N,N-di Isopropylethylamine (63 mg, 0.49 mmol). Subsequently, 1-propyl phosphoric anhydride (125 mg, 0.392 mmol) was added to the above solution, and reacted at room temperature for 18 hours.
加水淬灭反应，加乙酸乙酯(20毫升)稀释，水和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏，所得残余物用TLC板纯化得23毫克黄色固体(S)-3-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：38.0％)。MS(ESI)M/Z：[M+H] +＝616.20。 The reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml × 3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by TLC plate to obtain 23 mg of yellow solid ( S)-3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-benzene Propyl propionamido) tert-butyl benzoate (yield: 38.0%). MS(ESI) M/Z: [M+H] + =616.20.
步骤B：合成(S)-3-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸Step B: Synthesis of (S)-3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)benzoic acid
Figure PCTCN2020130361-appb-000046
Figure PCTCN2020130361-appb-000046
将(S)-3-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸叔丁酯(23.0毫克，0.037毫摩尔)溶于二氯甲烷(3.0毫升)。随后，向上述溶液中加入三氟乙酸(0.6毫升)。室温反应3小时。(S)-3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido) tert-butyl benzoate (23.0 mg, 0.037 mmol) was dissolved in dichloromethane (3.0 mL). Subsequently, trifluoroacetic acid (0.6 mL) was added to the above solution. React at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，纯化得13.5毫克黄色固体(S)-3-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)苯甲酸(收率：64.0％)。MS(ESI)M/Z：[M+H] +＝560.13。 1H NMR(400MHz,DMSO)δ12.99(s,1H),10.40(s,1H),9.75(s,1H),8.26(s,1H),7.93(d,J＝2.2Hz,1H),7.80(d,J＝8.6Hz,2H),7.75(dd,J＝8.6,2.2Hz,1H),7.66(d,J＝7.7Hz,1H),7.45(t,J＝7.9Hz,1H),7.28(dq,J＝15.6,7.3Hz,5H),5.32(s,1H),4.19–3.94(m,2H),3.72(s,2H),3.36(s,1H),3.10(dd,J＝14.4,10.2Hz,1H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, 13.5 mg of yellow solid (S)-3-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper was obtained after purification (Azin-1-yl)-3-phenylpropionamido)benzoic acid (yield: 64.0%). MS(ESI) M/Z: [M+H] + =560.13. 1 H NMR (400MHz, DMSO) δ 12.99 (s, 1H), 10.40 (s, 1H), 9.75 (s, 1H), 8.26 (s, 1H), 7.93 (d, J = 2.2 Hz, 1H), 7.80(d,J=8.6Hz,2H),7.75(dd,J=8.6,2.2Hz,1H),7.66(d,J=7.7Hz,1H),7.45(t,J=7.9Hz,1H), 7.28(dq,J=15.6,7.3Hz,5H), 5.32(s,1H), 4.19–3.94(m,2H), 3.72(s,2H), 3.36(s,1H), 3.10(dd,J= 14.4, 10.2 Hz, 1H).
实施例7Example 7
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid
Figure PCTCN2020130361-appb-000047
Figure PCTCN2020130361-appb-000047
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000048
Figure PCTCN2020130361-appb-000048
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(50毫克，0.11毫摩尔)，5-氨基-1H-苯并[d]咪唑-2-羧酸叔丁酯(32毫克，0.14毫摩尔)溶于N,N-二甲基甲酰胺(2.0毫升)中，加入N,N-二异丙基乙胺(72毫克，0.56毫摩尔)。随后，向上述溶液中加入1-丙基磷酸酐(143毫克，0.45毫摩尔),室温反应18小时。(S)-2-(4-(5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenyl Propionic acid (50 mg, 0.11 mmol), 5-amino-1H-benzo[d]imidazole-2-carboxylic acid tert-butyl ester (32 mg, 0.14 mmol) dissolved in N,N-dimethylformamide (2.0 mL) was added N,N-diisopropylethylamine (72 mg, 0.56 mmol). Subsequently, 1-propyl phosphoric anhydride (143 mg, 0.45 mmol) was added to the above solution, and reacted at room temperature for 18 hours.
加水淬灭反应，加乙酸乙酯(20毫升)稀释，水和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏，所得残余物用TLC板纯化(乙酸乙酯：正己烷＝2:1)得36毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸叔丁酯(收率：48.0％)。MS(ESI)M/Z：[M+H] +＝656.20。 The reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml×3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue obtained was purified with a TLC plate (ethyl acetate: n-hexane). Alkane=2:1) to give 36 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-diox Piperazin-1-yl)-3-phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid tert-butyl ester (yield: 48.0%). MS(ESI) M/Z: [M+H] + =656.20.
步骤B:合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid
Figure PCTCN2020130361-appb-000049
Figure PCTCN2020130361-appb-000049
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸叔丁酯(36毫克，0.055毫摩尔)溶于二氯甲烷(3.0毫升)。随后，向上述溶液中加入三氟乙酸(0.6毫升)。室温反应3小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid tert-butyl ester (36 mg, 0.055 mmol) was dissolved in dichloromethane (3.0 mL). Subsequently, trifluoroacetic acid (0.6 mL) was added to the above solution. React at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，纯化得20毫克浅黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸(收率：62.0％)。MS(ESI)M/Z：[M+H] +＝600.14。 1H NMR(400 MHz,MeOD)δ9.45(s,1H),8.09(d,J＝64.0Hz,3H),7.77(s,1H),7.70(d,J＝8.1Hz,2H),7.56(s,1H),7.32(ddd,J＝24.0,15.8,7.9Hz,6H),5.42(d,J＝24.7Hz,1H),5.35(t,J＝4.6Hz,1H),4.18(d,J＝56.4Hz,2H),3.83(s,2H),3.44(dd,J＝14.2,6.7Hz,1H),3.22–3.12(m,1H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, 20 mg of light yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxo were obtained. Piperazin-1-yl)-3-phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid (yield: 62.0%). MS(ESI) M/Z: [M+H] + =600.14. 1 H NMR(400 MHz,MeOD)δ9.45(s,1H), 8.09(d,J=64.0Hz,3H),7.77(s,1H),7.70(d,J=8.1Hz,2H),7.56 (s, 1H), 7.32 (ddd, J = 24.0, 15.8, 7.9 Hz, 6H), 5.42 (d, J = 24.7 Hz, 1H), 5.35 (t, J = 4.6 Hz, 1H), 4.18 (d, J = 56.4 Hz, 2H), 3.83 (s, 2H), 3.44 (dd, J = 14.2, 6.7 Hz, 1H), 3.22-3.12 (m, 1H).
实施例8Example 8
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000050
Figure PCTCN2020130361-appb-000050
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000051
Figure PCTCN2020130361-appb-000051
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(41毫克，0.093毫摩尔)，5-氨基-1H-吲哚-2-羧酸叔丁酯(26毫克，0.11毫摩尔)溶于N,N-二甲基甲酰胺(2.0毫升)中，加入N,N-二异丙基乙胺(60毫克，0.46毫摩尔)。随后，向上述溶液中加入1-丙基磷酸酐(118毫克，0.37毫摩尔),室温反应18小时。(S)-2-(4-(5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-phenyl Propionic acid (41 mg, 0.093 mmol), tert-butyl 5-amino-1H-indole-2-carboxylate (26 mg, 0.11 mmol) dissolved in N,N-dimethylformamide (2.0 mL) Add N,N-diisopropylethylamine (60 mg, 0.46 mmol). Subsequently, 1-propyl phosphoric anhydride (118 mg, 0.37 mmol) was added to the above solution, and reacted at room temperature for 18 hours.
加水淬灭反应，加乙酸乙酯(20毫升)稀释，水和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏，所得残余物用TLC板纯化(乙酸乙酯：正己烷＝3:1)得45毫克浅黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：73.0％)。MS(ESI)M/Z：[M+H] +＝655.21。 The reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml×3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue obtained was purified with a TLC plate (ethyl acetate: n-hexane). Alkane=3:1) to give 45 mg of light yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-di (Oxopiperazin-1-yl)-3-phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 73.0%). MS(ESI) M/Z: [M+H] + =655.21.
步骤B:合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000052
Figure PCTCN2020130361-appb-000052
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(45毫克，0.068毫摩尔)溶于二氯甲烷(3.0毫升)。随后，向上述溶液中加入三氟乙酸(0.6毫升)。室温反应3小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (45 mg, 0.068 mmol) was dissolved in dichloromethane (3.0 mL). Subsequently, trifluoroacetic acid (0.6 mL) was added to the above solution. React at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，纯化得30毫克浅黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：73.0％)。MS(ESI)M/Z：[M+H] +＝599.14。 1H NMR(500MHz,DMSO)δ12.91(s,1H),11.71(s,1H),10.14(d,J＝9.5Hz,1H),9.75(s,1H),7.99(s,1H),7.93(d,J＝2.2Hz,1H),7.80(d,J＝8.6Hz,1H),7.78–7.71(m,1H),7.38–7.22(m,7H),7.05(s,1H),5.32(dd,J＝12.8,7.3Hz,1H),3.97(s,2H),3.71(s,2H),3.52–3.34(m,1H),3.09(dd,J＝14.0,10.6Hz,1H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, 30 mg of light yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxo was obtained Piperazin-1-yl)-3-phenylpropionamido)-1H-indole-2-carboxylic acid (yield: 73.0%). MS (ESI) M/Z: [M+H] + =599.14. 1 H NMR (500MHz, DMSO) δ 12.91 (s, 1H), 11.71 (s, 1H), 10.14 (d, J = 9.5 Hz, 1H), 9.75 (s, 1H), 7.99 (s, 1H), 7.93(d,J=2.2Hz,1H),7.80(d,J=8.6Hz,1H),7.78–7.71(m,1H),7.38–7.22(m,7H),7.05(s,1H),5.32 (dd, J=12.8, 7.3 Hz, 1H), 3.97 (s, 2H), 3.71 (s, 2H), 3.52-3.34 (m, 1H), 3.09 (dd, J=14.0, 10.6 Hz, 1H).
实施例9Example 9
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000053
Figure PCTCN2020130361-appb-000053
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000054
Figure PCTCN2020130361-appb-000054
将(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(119毫克，0.18毫摩尔)，哌啶-1-碳酰氯(28毫克，0.19毫摩尔)溶于二氯甲烷(5.0毫升)中，加入三乙胺(53毫克，0.53毫摩尔)。室温反应18小时。Add (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-diox Piperazine-1-yl) propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (119 mg, 0.18 mmol), piperidine-1-carbonyl chloride (28 mg, 0.19 (Mmol) was dissolved in dichloromethane (5.0 mL), and triethylamine (53 mg, 0.53 mmol) was added. React at room temperature for 18 hours.
加水淬灭反应，加乙酸乙酯(20毫升)稀释，水和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏，所得残余物用TLC板纯化得71毫克浅黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸叔丁酯(收率：53.0％)。MS(ESI)M/Z：[M+H] +＝881.34。 The reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml × 3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by TLC plate to obtain 71 mg of light yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid tert-butyl ester (yield: 53.0%). MS (ESI) M/Z: [M+H] + =881.34.
步骤B:合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000055
Figure PCTCN2020130361-appb-000055
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(71毫克，0.08毫摩尔)溶于二氯甲烷(3.0毫升)。随后，向上述溶液中加入三氟乙酸(0.6毫升)。室温反应3小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (71 mg, 0.08 mmol) dissolved in dichloro Methane (3.0 mL). Subsequently, trifluoroacetic acid (0.6 mL) was added to the above solution. React at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，纯化得30.5毫克类白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：38.0％)。MS(ESI)M/Z：[M+H] +＝725.22。 1H NMR(500MHz,DMSO)δ12.90(s,1H),11.70(s,1H),10.11(s,1H),9.78(s,1H),8.37(s,1H),8.00(s,1H),7.95(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.3Hz,1H),7.36(ddd,J＝11.7,10.7,5.2Hz,4H),7.15(t,J＝11.2Hz,2H),7.06(d,J＝1.7Hz,1H),5.30(dd,J＝9.8,5.8Hz,1H),4.05–3.83(m,2H),3.70(d,J＝31.6Hz,2H),3.55(s,4H),3.23(dd,J＝14.2,5.3Hz,1H),3.01(dd,J＝14.0,10.1Hz,1H),1.61–1.53(m,2H),1.53–1.45(m,4H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, 30.5 mg of off-white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxo was obtained. Piperazin-1-yl)-3-(4-(piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 38.0%). MS (ESI) M/Z: [M+H] + =725.22. 1 H NMR (500MHz, DMSO) δ 12.90 (s, 1H), 11.70 (s, 1H), 10.11 (s, 1H), 9.78 (s, 1H), 8.37 (s, 1H), 8.00 (s, 1H) ), 7.95 (d, J = 2.2 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.76 (dd, J = 8.6, 2.3 Hz, 1H), 7.36 (ddd, J = 11.7, 10.7, 5.2Hz, 4H), 7.15 (t, J = 11.2 Hz, 2H), 7.06 (d, J = 1.7 Hz, 1H), 5.30 (dd, J = 9.8, 5.8 Hz, 1H), 4.05-3.83 (m, 2H), 3.70 (d, J = 31.6 Hz, 2H), 3.55 (s, 4H), 3.23 (dd, J = 14.2, 5.3 Hz, 1H), 3.01 (dd, J = 14.0, 10.1 Hz, 1H), 1.61–1.53(m,2H), 1.53–1.45(m,4H).
实施例10Example 10
合成(S)-3-氯-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-3-chloro-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000056
Figure PCTCN2020130361-appb-000056
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成3-氯-5-硝基-1H-吲哚-2-羧酸Step A: Synthesis of 3-chloro-5-nitro-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000057
Figure PCTCN2020130361-appb-000057
将5-硝基-1H-吲哚-2-羧酸(500毫克，2.43毫摩尔)，N-氯代丁二酰胺(357毫克，2.67毫摩尔)溶解于N,N-二甲基甲酰胺(4毫升)，室温搅拌4小时。Dissolve 5-nitro-1H-indole-2-carboxylic acid (500 mg, 2.43 mmol) and N-chlorosuccinamide (357 mg, 2.67 mmol) in N,N-dimethylformamide (4 mL), stirring at room temperature for 4 hours.
将反应液用乙酸乙酯(100毫升)稀释，分别用水(20毫升×2次)和饱和食盐水(20毫升)，有机相用无水硫酸钠干燥，过滤浓缩，所得粗品3-氯-5-硝基-1H-吲哚-2-羧酸直接用于下一步反应。LCMS:RT＝3.39min,[M-H] -＝239.03。 The reaction solution was diluted with ethyl acetate (100 ml), respectively, with water (20 ml × 2 times) and saturated brine (20 ml), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain crude 3-chloro-5 -Nitro-1H-indole-2-carboxylic acid was directly used in the next reaction. LCMS: RT = 3.39 min, [MH] - =239.03.
步骤B：合成3-氯-5-硝基-1H-吲哚-1,2-二羧酸二叔丁酯Step B: Synthesis of 3-chloro-5-nitro-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000058
Figure PCTCN2020130361-appb-000058
将粗品3-氯-5-硝基-1H-吲哚-2-羧酸(3.24毫摩尔)，二碳酸二叔丁酯(3.18克，14.58毫摩尔)溶解于N,N-二甲基甲酰胺(3毫升),0摄氏度条件下加入4-二甲氨基吡啶(357毫克，2.92毫摩尔)，室温搅拌过夜。The crude 3-chloro-5-nitro-1H-indole-2-carboxylic acid (3.24 mmol), di-tert-butyl dicarbonate (3.18 g, 14.58 mmol) were dissolved in N,N-dimethylformaldehyde Amide (3 mL), 4-dimethylaminopyridine (357 mg, 2.92 mmol) was added at 0 degrees Celsius, and stirred at room temperature overnight.
将反应液用冰淬灭，用乙酸乙酯(100毫升)稀释，分别用水(20毫升×2次)和饱和食盐水(20毫升)，有机相用无水硫酸钠干燥，过滤浓缩，所得粗品3-氯-5-硝基-1H-吲哚-1,2-二羧酸二叔丁酯直接用于下一步反应。LCMS:RT＝5.27min,[M-56-H] -＝396.11。 The reaction solution was quenched with ice, diluted with ethyl acetate (100 mL), water (20 mL×2 times) and saturated brine (20 mL) respectively, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product 3-chloro-5-nitro-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester was directly used in the next reaction. LCMS: RT = 5.27 min, [M-56-H] - = 396.11.
步骤C：合成3-氯-5-胺基-1H-吲哚-1,2-二羧酸二叔丁酯Step C: Synthesis of 3-chloro-5-amino-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000059
Figure PCTCN2020130361-appb-000059
将粗品3-氯-5-硝基-1H-吲哚-1,2-二羧酸二叔丁酯(3.24毫摩尔)和钯碳(60毫克)溶解于乙醇(50毫升)，用氢气球提供氢气源，室温条件下反应3小时。Dissolve the crude 3-chloro-5-nitro-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (3.24 mmol) and palladium on carbon (60 mg) in ethanol (50 ml), and use a hydrogen balloon Provide a hydrogen source and react for 3 hours at room temperature.
将反应液用乙醇(100毫升)稀释，硅藻土过滤，滤饼用乙醇(100毫升)冲洗，有机相浓缩，所得粗品用硅胶柱层析纯化(石油醚：乙酸乙酯＝8/1)。得到220毫克白色固体3-氯-5-胺基-1H-吲哚-1,2-二羧酸二叔丁酯(三步总收率：18.6％)。LCMS:RT＝4.43min,[M+H] +＝367.19。 The reaction solution was diluted with ethanol (100 ml), filtered through Celite, the filter cake was washed with ethanol (100 ml), the organic phase was concentrated, and the crude product obtained was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8/1) . Obtained 220 mg of white solid 3-chloro-5-amino-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (three-step total yield: 18.6%). LCMS: RT = 4.43 min, [M+H] + =367.19.
步骤D：合成(S)-3-氯-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺)-1H-吲哚-2-羧酸叔丁酯Step D: Synthesis of (S)-3-chloro-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine -1-yl)-3-phenylpropionamide)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000060
Figure PCTCN2020130361-appb-000060
氮气保护条件下将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(47毫克，0.107毫摩尔)，3-氯-5-胺基-1H-吲哚-1,2-二羧酸二叔丁酯(79毫克，0.214毫摩尔)，1-丙基磷酸酐(171毫克，0.535毫摩尔)和N,N- 二异丙基乙胺(56微升，0.321毫摩尔)溶解于N,N-二甲基甲酰胺(1毫升)，室温搅拌过夜。(S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- 3-Phenylpropionic acid (47 mg, 0.107 mmol), di-tert-butyl 3-chloro-5-amino-1H-indole-1,2-dicarboxylate (79 mg, 0.214 mmol), 1 -Propyl phosphoric anhydride (171 mg, 0.535 mmol) and N,N-diisopropylethylamine (56 μl, 0.321 mmol) were dissolved in N,N-dimethylformamide (1 mL) at room temperature Stir overnight.
将反应液用乙酸乙酯(100毫升)稀释，分别用水(20毫升×2次)和饱和食盐水(20毫升)，有机相用无水硫酸钠干燥，过滤浓缩，所得粗品用硅胶柱层析纯化(石油醚：乙酸乙酯＝1/1)。得到50毫克黄色固体(S)-3-氯-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺)-1H-吲哚-2-羧酸叔丁酯(收率：68.0％)。LCMS:RT＝4.55min,[M+H] +＝689.39。 The reaction solution was diluted with ethyl acetate (100 ml), respectively, with water (20 ml × 2 times) and saturated brine (20 ml), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product obtained was subjected to silica gel column chromatography Purification (petroleum ether: ethyl acetate = 1/1). Obtain 50 mg of yellow solid (S)-3-chloro-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-phenylpropionamide)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 68.0%). LCMS: RT = 4.55 min, [M+H] + = 689.39.
步骤E：合成(S)-3-氯-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Step E: Synthesis of (S)-3-chloro-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine -1-yl)-3-phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000061
Figure PCTCN2020130361-appb-000061
将(S)-3-氯-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺)-1H-吲哚-2-羧酸叔丁酯(50毫克，0.073毫摩尔)溶解于二氯甲烷(5毫升)，室温条件下缓慢滴加三氟乙酸(1毫升)，室温条件下继续搅拌反应1.5小时。Add (S)-3-chloro-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-phenylpropionamide)-1H-indole-2-carboxylic acid tert-butyl ester (50 mg, 0.073 mmol) was dissolved in dichloromethane (5 ml), and trifluoroacetic acid was slowly added dropwise at room temperature (1 ml), continue to stir and react for 1.5 hours at room temperature.
将反应液用旋转蒸发仪浓缩，待用油泵进一步抽干后用甲醇溶解，然后向溶液体系中滴加正己烷，有大量固体析出，室温继续搅拌1小时，过滤得7.1毫克棕色固体(S)-3-氯-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：15.5％)。LCMS：RT＝3.67min，[M-H] -＝631.27。 1H NMR(500MHz,DMSO)δ13.36(s,1H),11.99(s,1H),10.26(s,1H),9.75(s,1H),8.06(s,1H),7.94(d,J＝2.2Hz,1H),7.81(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.2Hz,1H),7.42(s,2H),7.36–7.27(m,4H),7.24(t,J＝7.2Hz,1H),5.35–5.30(m,1H),3.11(dd,J＝14.3,10.2Hz,1H),2.05–1.94(m,1H)。 The reaction solution was concentrated with a rotary evaporator, and then it was further drained by an oil pump and dissolved in methanol. Then, n-hexane was added dropwise to the solution system. A large amount of solids precipitated. Continue stirring at room temperature for 1 hour, and filter to obtain 7.1 mg of brown solids (S) -3-chloro-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid (yield: 15.5%). LCMS: RT = 3.67 min, [MH] - =631.27. 1 H NMR (500MHz, DMSO) δ 13.36 (s, 1H), 11.99 (s, 1H), 10.26 (s, 1H), 9.75 (s, 1H), 8.06 (s, 1H), 7.94 (d, J =2.2Hz,1H),7.81(d,J=8.6Hz,1H),7.76(dd,J=8.6,2.2Hz,1H),7.42(s,2H),7.36-7.27(m,4H),7.24 (t, J = 7.2 Hz, 1H), 5.35-5.30 (m, 1H), 3.11 (dd, J = 14.3, 10.2 Hz, 1H), 2.05-1.94 (m, 1H).
实施例11Example 11
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1-甲基-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1-methyl-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000062
Figure PCTCN2020130361-appb-000062
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成1-甲基-5-硝基-1H-吲哚-2-羧酸乙酯Step A: Synthesis of 1-methyl-5-nitro-1H-indole-2-carboxylic acid ethyl ester
Figure PCTCN2020130361-appb-000063
Figure PCTCN2020130361-appb-000063
将5-硝基-1H-吲哚-2-羧酸乙酯(2.5克,10.7毫摩尔)和碳酸钾(2.9克,21.4毫摩尔)溶于N,N-二甲基甲酰胺(20.0毫升)中。然后将碘甲烷(2.3克,16.0毫摩尔)加入反应液中,加热升温到60℃并恒温搅拌4小时后,LCMS监测至反应完全,将反应液冷却至室温。Dissolve 5-nitro-1H-indole-2-carboxylic acid ethyl ester (2.5 g, 10.7 mmol) and potassium carbonate (2.9 g, 21.4 mmol) in N,N-dimethylformamide (20.0 ml )in. Then methyl iodide (2.3 g, 16.0 mmol) was added to the reaction solution, heated to 60° C. and stirred at constant temperature for 4 hours, LCMS monitored until the reaction was complete, and the reaction solution was cooled to room temperature.
向反应液中加饱和氯化铵水溶液淬灭,混合液用乙酸乙酯(40毫升×3次)萃取,合并有机相,有机相用饱和食盐水(30毫升×3次),无水硫酸钠干燥,减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/石油醚＝2/1)得到2.6克黄色固体1-甲基-5-硝基-1H-吲哚-2-羧酸乙酯(收率：98.1％)。LCMS：RT＝4.25min,[M+H] +＝249.02。 The reaction solution was quenched by adding saturated aqueous ammonium chloride solution, the mixed solution was extracted with ethyl acetate (40 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (30 ml × 3 times), anhydrous sodium sulfate Dry and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/petroleum ether = 2/1) to obtain 2.6 g of yellow solid 1-methyl-5-nitro-1H-indole-2-carboxylic acid ethyl Esters (yield: 98.1%). LCMS: RT = 4.25 min, [M+H] + =249.02.
步骤B：合成1-甲基-5-硝基-1H-吲哚-2-羧酸Step B: Synthesis of 1-methyl-5-nitro-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000064
Figure PCTCN2020130361-appb-000064
将1-甲基-5-硝基-1H-吲哚-2-羧酸乙酯(2.0克,8.1毫摩尔)溶于四氢呋喃(60.0毫升)中，然后将氢氧化钠溶液(2M,10.0毫升)加入反应液中,室温下搅拌4小时后,LCMS监测至反应完全。Ethyl 1-methyl-5-nitro-1H-indole-2-carboxylate (2.0 g, 8.1 mmol) was dissolved in tetrahydrofuran (60.0 ml), and then sodium hydroxide solution (2M, 10.0 ml ) Was added to the reaction solution, and after stirring for 4 hours at room temperature, LCMS monitored until the reaction was complete.
反应液减压浓缩,稀盐酸调节pH至3,析出大量固体,过滤,滤饼用水(30毫升×3次)洗涤，收集滤饼，干燥后得到1.6克白色固体1-甲基-5-硝基-1H-吲哚-2-羧酸(收率：89.9％)。LCMS：RT＝3.85min,[M+H] +＝219.04。 The reaction solution was concentrated under reduced pressure, adjusted to pH 3 with dilute hydrochloric acid, a large amount of solid was precipitated, filtered, the filter cake was washed with water (30 ml×3 times), the filter cake was collected and dried to obtain 1.6 g of white solid 1-methyl-5-nitro -1H-indole-2-carboxylic acid (yield: 89.9%). LCMS: RT = 3.85 min, [M+H] + =219.04.
步骤C：合成1-甲基-5-硝基-1H-吲哚-2-羧酸叔丁酯Step C: Synthesis of 1-methyl-5-nitro-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000065
Figure PCTCN2020130361-appb-000065
将1-甲基-5-硝基-1H-吲哚-2-羧酸(440毫克,2.0毫摩尔)溶于N,N-二甲基甲酰胺(4.0毫升)中，然后将CDI(486毫克,3.0毫摩尔)加入反应液中,室温下搅拌1小时后,将叔丁醇(371毫克,5.0毫摩尔)和1,8-二氮杂双环[5.4.0]十一碳-7-烯(396毫克,2.6毫摩尔)加入反应液中,室温下搅拌2小时后,LCMS监测至反应完全。1-Methyl-5-nitro-1H-indole-2-carboxylic acid (440 mg, 2.0 mmol) was dissolved in N,N-dimethylformamide (4.0 mL), and then CDI (486 Mg, 3.0 mmol) was added to the reaction solution, and after stirring for 1 hour at room temperature, tert-butanol (371 mg, 5.0 mmol) and 1,8-diazabicyclo[5.4.0]undec-7- Alkene (396 mg, 2.6 mmol) was added to the reaction solution, and after stirring for 2 hours at room temperature, LCMS monitored until the reaction was complete.
向反应液中加饱和氯化铵水溶液淬灭,混合液用乙酸乙酯(40毫升×3次)萃取,合并有机相,有机相用饱和食盐水(30毫升×3次),无水硫酸钠干燥,减压浓缩得到402毫克黄色固体1-甲基-5-硝基-1H-吲哚-2-羧酸叔丁酯(收率：72.8％)。LCMS：RT＝4.59min,[M+K] +＝316.91。 The reaction solution was quenched by adding saturated aqueous ammonium chloride solution, the mixed solution was extracted with ethyl acetate (40 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (30 ml × 3 times), anhydrous sodium sulfate It was dried and concentrated under reduced pressure to obtain 402 mg of yellow solid 1-methyl-5-nitro-1H-indole-2-carboxylic acid tert-butyl ester (yield: 72.8%). LCMS: RT = 4.59 min, [M+K] + = 316.91.
步骤D：合成1-甲基-5-氨基-1H-吲哚-2-羧酸叔丁酯Step D: Synthesis of 1-methyl-5-amino-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000066
Figure PCTCN2020130361-appb-000066
氮气保护下,将1-甲基-5-硝基-1H-吲哚-2-羧酸叔丁酯(402毫克,1.5毫摩尔)溶于乙醇(8.0毫升)和乙酸乙酯(4.0毫升)中,向反应液中加入钯碳(80毫克,20％),置换氢气后,室温下搅拌9小时。Under nitrogen protection, dissolve 1-methyl-5-nitro-1H-indole-2-carboxylic acid tert-butyl ester (402 mg, 1.5 mmol) in ethanol (8.0 mL) and ethyl acetate (4.0 mL) In the reaction solution, palladium-carbon (80 mg, 20%) was added to the reaction solution, and after replacing hydrogen, the mixture was stirred at room temperature for 9 hours.
反应液垫硅藻土过滤,滤饼用乙酸乙酯(20毫升×3次)洗涤。合并滤液及洗涤液,减压浓缩,所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/2)。得到317毫克黄色固体1-甲基-5-氨基-1H-吲哚-2-羧酸叔丁酯(收率：73.0％)。LCMS：RT＝2.83min,[M+H] +＝247.17。 The reaction solution was filtered through Celite, and the filter cake was washed with ethyl acetate (20 ml×3 times). The filtrate and washings were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/2). Obtained 317 mg of yellow solid 1-methyl-5-amino-1H-indole-2-carboxylic acid tert-butyl ester (yield: 73.0%). LCMS: RT = 2.83 min, [M+H] + =247.17.
步骤E：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-硝基苯基)丙酰氨基)-1-甲基-1H-吲哚-2-羧酸叔丁酯Step E: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-nitrophenyl)propionylamino)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000067
Figure PCTCN2020130361-appb-000067
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸(263毫克,0.5毫摩尔)和N,N-二异丙基乙胺(0.2毫升,1.5毫摩尔)溶于N,N-二甲基甲酰胺(3.0毫升)中。随后,向上述溶液中加入2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(380毫克,1.0毫摩尔)和1-甲基-5-氨基-1H-吲哚-2-羧酸叔丁酯(123.0毫克,0.5毫摩尔)。在室温下搅拌过夜。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -Nitrophenyl) propionic acid (263 mg, 0.5 mmol) and N,N-diisopropylethylamine (0.2 mL, 1.5 mmol) dissolved in N,N-dimethylformamide (3.0 mL) in. Subsequently, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (380 mg, 1.0 mmol) and 1- Tert-butyl methyl-5-amino-1H-indole-2-carboxylate (123.0 mg, 0.5 mmol). Stir overnight at room temperature.
向反应液中加水淬灭反应。混合液用乙酸乙酯(20毫升×3次)萃取。合并有机相,有机相先用饱和食盐水(10毫升×3次),然后用无水硫酸钠干燥,最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/1)。得到230毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-硝基苯基)丙酰氨基)-1-甲基-1H-吲哚-2-羧酸叔丁酯(收率：60.0％)。LCMS：RT＝4.24min,[M-H] -＝712.28。 The reaction was quenched by adding water to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). The organic phase was combined, and the organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/1). Obtain 230 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- (Yl)-3-(-4-nitrophenyl)propionamido)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester (yield: 60.0%). LCMS: RT = 4.24 min, [MH] - =712.28.
步骤F：合成(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1-甲基-1H-吲哚-2-羧酸叔丁酯Step F: Synthesis of (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3 -Dioxopiperazin-1-yl)propionylamino)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000068
Figure PCTCN2020130361-appb-000068
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-硝基苯基)丙酰氨基)-1-甲基-1H-吲哚-2-羧酸叔丁酯(230毫克,0.3毫摩尔)溶于甲醇(8.0毫升)和冰醋酸(4.0毫升)中。随后,向上述溶液中加入还原性铁粉(168毫克,3.0毫摩尔),在70℃下搅拌4小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(-4-nitrophenyl)propionamido)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester (230 mg, 0.3 mmol) dissolved in methanol (8.0 ml) and glacial acetic acid (4.0 mL). Subsequently, reducing iron powder (168 mg, 3.0 mmol) was added to the above solution and stirred at 70°C for 4 hours.
反应液冷却至室温。加碳酸氢钠中合,混合液用乙酸乙酯(20毫升×3次)萃取。合并有机相,有机相先用饱和食盐水(10毫升×3次),无水硫酸钠干燥,减压浓缩得到205毫克黄色固体(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1-甲基-1H-吲哚-2-羧酸叔丁酯(收率93.0％)。LCMS：RT＝4.23min,[M-H] -＝684.12。 The reaction solution was cooled to room temperature. Add sodium bicarbonate to neutralize, and extract the mixture with ethyl acetate (20 ml×3 times). Combine the organic phases, first use saturated brine (10 ml × 3 times), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 205 mg of yellow solid (S)-5-(3-(4-aminophenyl)- 2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1-methyl- 1H-indole-2-carboxylic acid tert-butyl ester (yield 93.0%). LCMS: RT = 4.23 min, [MH] - =684.12.
步骤G：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-(哌啶-1-甲酰氨基)苯基)丙酰氨基)-1-甲基-1H-吲哚-2-羧酸叔丁酯Step G: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-(piperidine-1-carboxamido)phenyl)propionamido)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000069
Figure PCTCN2020130361-appb-000069
将(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1-甲基-1H-吲哚-2-羧酸叔丁酯(205毫克,0.3毫摩尔)和三乙胺(0.1毫升,0.75毫摩尔)溶于二氯甲烷(5.0毫升)中。于冰浴中向反应液中加入哌啶-1-羰基氯(53毫克,0.36毫摩尔),在室温下搅拌2小时。Add (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-diox Piperazin-1-yl) propionylamino)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester (205 mg, 0.3 mmol) and triethylamine (0.1 mL, 0.75 mmol) Dissolve in dichloromethane (5.0 mL). Piperidine-1-carbonyl chloride (53 mg, 0.36 mmol) was added to the reaction solution in an ice bath, and the mixture was stirred at room temperature for 2 hours.
向反应液中加入饱和碳酸氢钠溶液(10毫升)淬灭反应。混合液用乙酸乙酯(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次),然后用无水硫酸钠干燥,最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/甲醇＝10/1)。得到130毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-(哌啶-1-甲酰氨基)苯基)丙酰氨基)-1-甲基-1H-吲哚-2-羧酸叔丁酯(收率：38％)。LCMS：RT＝4.19min,[M+H] +＝795.23。 Saturated sodium bicarbonate solution (10 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/methanol=10/1). Obtain 130 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-(piperidine-1-carboxamido)phenyl)propionylamino)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester (Yield: 38% ). LCMS: RT = 4.19 min, [M+H] + =795.23.
步骤H：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1-甲基-1H-吲哚-2-羧酸Step H: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1-methyl-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000070
Figure PCTCN2020130361-appb-000070
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-(哌啶-1-甲酰氨基)苯基)丙酰氨基)-1-甲基-1H-吲哚-2-羧酸叔丁酯(90毫克,0.11毫摩尔)溶于二氯甲烷(5.0毫升)中。随后,向上述溶液中加入三氟乙酸(1.0毫升),在室温下搅拌1小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(-4-(piperidine-1-carboxamido)phenyl)propionamido)-1-methyl-1H-indole-2-carboxylic acid tert-butyl ester (90 mg, 0.11 mmol) dissolved in Methylene chloride (5.0 mL). Subsequently, trifluoroacetic acid (1.0 ml) was added to the above solution and stirred at room temperature for 1 hour.
将反应液空气浴中减压浓缩。将所得残余物用制备型高效液相色谱纯化,得到29毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1-甲基-1H-吲哚-2-羧酸(收率：35.0％)。LCMS：RT＝3.66min,[M-H] -＝737.14。 1H NMR(500MHz,DMSO)δ12.89(s,1H),10.16(s,1H),9.78(s,1H),8.38(s,1H),8.03(s,1H),7.95(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.3Hz,1H),7.53(d,J＝9.0Hz,1H),7.45–7.38(m,2H),7.18(d,J＝6.4Hz,1H),7.14(d,J＝8.2Hz,1H),5.75(s,1H),5.35–5.27(m,1H),4.00(s,2H),3.43–3.38(m,3H),3.23(d,J＝14.1Hz,1H),3.06–2.97(m,1H),2.55–2.53(m,2H),1.56(d,J＝5.3Hz,2H),1.48(d,J＝4.1Hz,4H)。 The reaction solution was concentrated under reduced pressure in an air bath. The resulting residue was purified by preparative high performance liquid chromatography to obtain 29 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl) -2,3-dioxopiperazin-1-yl)-3-(4-(piperidine-1-carboxamido)phenyl)propionamido)-1-methyl-1H-indole-2 -Carboxylic acid (yield: 35.0%). LCMS: RT = 3.66 min, [MH] - =737.14. 1 H NMR (500MHz, DMSO) δ 12.89 (s, 1H), 10.16 (s, 1H), 9.78 (s, 1H), 8.38 (s, 1H), 8.03 (s, 1H), 7.95 (d, J =2.2Hz,1H),7.82(d,J=8.6Hz,1H),7.76(dd,J=8.6,2.3Hz,1H),7.53(d,J=9.0Hz,1H),7.45–7.38(m ,2H),7.18(d,J=6.4Hz,1H),7.14(d,J=8.2Hz,1H),5.75(s,1H),5.35-5.27(m,1H),4.00(s,2H) ,3.43–3.38(m,3H), 3.23(d,J=14.1Hz,1H),3.06–2.97(m,1H),2.55–2.53(m,2H),1.56(d,J=5.3Hz,2H ), 1.48 (d, J = 4.1 Hz, 4H).
实施例12Example 12
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000071
Figure PCTCN2020130361-appb-000071
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成哌啶-4-醇的三氟乙酸盐Step A: Synthesis of the trifluoroacetate salt of piperidin-4-ol
Figure PCTCN2020130361-appb-000072
Figure PCTCN2020130361-appb-000072
将4-羟基哌啶-1-羧酸叔丁酯(1.0克,5.0毫摩尔)溶于二氯甲烷(10.0毫升)中。随后,向上述溶液中加入三氟乙酸(2.0毫升),在室温下搅拌1小时。Tert-butyl 4-hydroxypiperidine-1-carboxylate (1.0 g, 5.0 mmol) was dissolved in dichloromethane (10.0 mL). Subsequently, trifluoroacetic acid (2.0 ml) was added to the above solution and stirred at room temperature for 1 hour.
将反应液空气浴中减压浓缩,得到1.0克黄色固体哌啶-4-醇的三氟乙酸盐粗品(收率：93.5％)。The reaction solution was concentrated under reduced pressure in an air bath to obtain 1.0 g of crude trifluoroacetate of piperidin-4-ol as a yellow solid (yield: 93.5%).
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-羟基哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Hydroxypiperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000073
Figure PCTCN2020130361-appb-000073
室温下,将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(178毫克,0.2毫摩尔)和哌啶-4-醇的三氟乙酸盐(860毫克,4.0毫摩尔)溶于四氢呋喃(6.0毫升)中。随后,向上述溶液中N,N-二异丙基乙胺(1.0毫升,6毫摩尔)。在60℃反应12小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (178 mg, 0.2 mmol) and piperidine The trifluoroacetate salt of pyridin-4-ol (860 mg, 4.0 mmol) was dissolved in tetrahydrofuran (6.0 mL). Subsequently, add N,N-diisopropylethylamine (1.0 mL, 6 mmol) to the above solution. React at 60°C for 12 hours.
向反应液中加水淬灭,混合液用乙酸乙酯(30毫升×3次)萃取,合并有机相,有机相用饱和食盐水(20毫升×3次),无水硫酸钠干燥,减压浓缩得到180毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-羟基哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯粗品(收率：100％)。LCMS：RT＝4.14min,[M-H] -＝867.42。 The reaction solution was quenched by adding water, the mixed solution was extracted with ethyl acetate (30 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (20 ml × 3 times), dried with anhydrous sodium sulfate, and concentrated under reduced pressure Obtain 180 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(4-hydroxypiperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl crude product (Yield: 100%). LCMS: RT = 4.14 min, [MH] - =867.42.
步骤C：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step C: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000074
Figure PCTCN2020130361-appb-000074
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-羟基哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(180毫克,0.2毫摩尔)溶于二氯甲烷(5.0毫升)中。随后,向上述溶液中加入三氟乙酸(1.0毫升),在室温下搅拌1小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Hydroxypiperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (180 mg, 0.2 mmol) In dichloromethane (5.0 mL). Subsequently, trifluoroacetic acid (1.0 ml) was added to the above solution and stirred at room temperature for 1 hour.
将反应液空气浴中减压浓缩。将所得残余物用制备型高效液相色谱纯化,得到27毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：18.2％)。LCMS：RT＝3.16min,[M-H] -＝739.23。 1H NMR(400MHz,DMSO)δ11.71(s,1H),10.13(s,1H),9.78(s,1H),8.43(s,1H),8.00(s,1H),7.95(d,J＝1.9Hz,1H),7.82(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.2Hz,1H),7.43–7.30(m,3H),7.14(d,J＝7.9Hz,1H),7.06(d,J＝1.6Hz,1H),5.30(dd,J＝9.5,5.6Hz,1H),3.82(d,J＝13.7Hz,2H),3.70–3.60(m,2H),3.47(m,6H),3.22(dd,J＝14.9,6.4Hz,2H),3.08–2.96(m,2H),2.54(s,1H),1.73(d,J＝8.9Hz,2H),1.36–1.21(m,2H)。 The reaction solution was concentrated under reduced pressure in an air bath. The resulting residue was purified by preparative high performance liquid chromatography to obtain 27 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl) -2,3-dioxopiperazin-1-yl)-3-(4-(4-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxy Acid (yield: 18.2%). LCMS: RT = 3.16 min, [MH] - =739.23. 1 H NMR (400MHz, DMSO) δ 11.71 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.43 (s, 1H), 8.00 (s, 1H), 7.95 (d, J =1.9Hz,1H),7.82(d,J=8.6Hz,1H),7.76(dd,J=8.6,2.2Hz,1H),7.43-7.30(m,3H),7.14(d,J=7.9Hz ,1H), 7.06(d,J=1.6Hz,1H), 5.30(dd,J=9.5,5.6Hz,1H), 3.82(d,J=13.7Hz,2H), 3.70–3.60(m,2H) ,3.47(m,6H),3.22(dd,J=14.9,6.4Hz,2H),3.08–2.96(m,2H),2.54(s,1H),1.73(d,J=8.9Hz,2H), 1.36–1.21(m,2H).
实施例13Example 13
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(1,1-二氧代四氢-1H-噻喃-4-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-(1,1-dioxotetrahydro-1H-thiopyran-4-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000075
Figure PCTCN2020130361-appb-000075
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(1,1-二氧代四氢-2H-噻喃-4-基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(1,1-dioxotetrahydro-2H-thiopyran-4-yl)ureido)phenyl)propionylamino)-1H-indole-1,2- Di-tert-butyl dicarboxylate
Figure PCTCN2020130361-appb-000076
Figure PCTCN2020130361-appb-000076
室温下,将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(178毫克，0.2毫摩尔)和4-氨基四氢-2H-噻喃1,1-二氧化物(371毫克，2.0毫摩尔)溶于四氢呋喃(6.0毫升)中。随后,向上述溶液中N,N-二异丙基乙胺(0.7毫升，4毫摩尔)。在60℃反应12小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (178 mg, 0.2 mmol) and 4 -Aminotetrahydro-2H-thiopyran 1,1-dioxide (371 mg, 2.0 mmol) was dissolved in tetrahydrofuran (6.0 mL). Then, add N,N-diisopropylethylamine (0.7 mL, 4 mmol) to the above solution. React at 60°C for 12 hours.
向反应液中加水淬灭,混合液用乙酸乙酯(30毫升×3次)萃取,合并有机相,有机相用饱和食盐水(20毫升×3次),无水硫酸钠干燥,减压浓缩得到180毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(1,1-二氧代四氢-2H-噻喃-4-基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：95.2％)。LCMS：RT＝4.17min,[M+H] +＝945.28。 The reaction solution was quenched by adding water, the mixed solution was extracted with ethyl acetate (30 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (20 ml × 3 times), dried with anhydrous sodium sulfate, and concentrated under reduced pressure Obtain 180 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-(1,1-dioxotetrahydro-2H-thiopyran-4-yl)ureido)phenyl)propionamido)-1H-indole-1,2 -Di-tert-butyl dicarboxylic acid (yield: 95.2%). LCMS: RT = 4.17 min, [M+H] + =945.28.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(1,1-二氧代四氢-1H-噻喃-4-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(1,1-dioxotetrahydro-1H-thiopyran-4-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000077
Figure PCTCN2020130361-appb-000077
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(1,1-二氧代四氢-2H-噻喃-4-基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(180毫克，0.19毫摩尔)溶于二氯甲烷(5.0毫升)中。随后,向上述溶液中加入三氟乙酸(1.0毫升),在室温下搅拌1小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-(1,1-dioxotetrahydro-2H-thiopyran-4-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester (180 mg, 0.19 mmol) was dissolved in dichloromethane (5.0 mL). Subsequently, trifluoroacetic acid (1.0 ml) was added to the above solution and stirred at room temperature for 1 hour.
将反应液空气浴中减压浓缩。将所得残余物用制备型高效液相色谱纯化,得到26毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(1,1-二氧代四氢-1H-噻喃-4-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧 酸(收率：17.2％)。LCMS：RT＝3.24min,[M-H] -＝787.13。 1H NMR(400MHz,DMSO)δ13.05–12.68(m,1H),11.71(s,1H),10.11(s,1H),9.78(s,1H),8.25(s,1H),7.99(s,1H),7.94(d,J＝2.3Hz,1H),7.82(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.3Hz,1H),7.42–7.30(m,3H),7.13(d,J＝8.1Hz,2H),7.06(d,J＝1.6Hz,1H),6.39(d,J＝7.5Hz,1H),5.75(s,1H),5.28(dd,J＝10.2,5.8Hz,1H),3.84(s,2H),3.73(s,2H),3.26–3.17(m,2H),3.05(d,J＝11.8Hz,2H),2.54(s,1H),2.12(s,2H),1.90(dd,J＝21.0,10.2Hz,2H). The reaction solution was concentrated under reduced pressure in an air bath. The resulting residue was purified by preparative high performance liquid chromatography to obtain 26 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl) -2,3-dioxopiperazin-1-yl)-3-(4-(3-(1,1-dioxotetrahydro-1H-thiopyran-4-yl)ureido)phenyl) Propanamido)-1H-indole-2-carboxylic acid (yield: 17.2%). LCMS: RT = 3.24 min, [MH] - =787.13. 1 H NMR (400MHz, DMSO) δ 13.05-12.68 (m, 1H), 11.71 (s, 1H), 10.11 (s, 1H), 9.78 (s, 1H), 8.25 (s, 1H), 7.99 (s ,1H),7.94(d,J=2.3Hz,1H),7.82(d,J=8.6Hz,1H),7.76(dd,J=8.6,2.3Hz,1H),7.42-7.30(m,3H) , 7.13 (d, J = 8.1 Hz, 2H), 7.06 (d, J = 1.6 Hz, 1H), 6.39 (d, J = 7.5 Hz, 1H), 5.75 (s, 1H), 5.28 (dd, J = 10.2,5.8Hz,1H),3.84(s,2H),3.73(s,2H),3.26-3.17(m,2H),3.05(d,J=11.8Hz,2H),2.54(s,1H), 2.12 (s, 2H), 1.90 (dd, J = 21.0, 10.2 Hz, 2H).
实施例14Example 14
合成5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000078
Figure PCTCN2020130361-appb-000078
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成1-氧杂-8-氮杂螺[4.5]癸-3-醇的三氟乙酸盐Step A: Synthesis of 1-oxa-8-azaspiro[4.5]dec-3-ol trifluoroacetate
Figure PCTCN2020130361-appb-000079
Figure PCTCN2020130361-appb-000079
将3-羟基-1-氧杂-8-氮杂螺[4-]癸烷-8-羧酸叔丁酯(1.0克，5.0毫摩尔)溶于二氯甲烷(10.0毫升)中。随后,向上述溶液中加入三氟乙酸(2.0毫升),在室温下搅拌1小时。Tert-butyl 3-hydroxy-1-oxa-8-azaspiro[4-]decane-8-carboxylate (1.0 g, 5.0 mmol) was dissolved in dichloromethane (10.0 mL). Subsequently, trifluoroacetic acid (2.0 ml) was added to the above solution and stirred at room temperature for 1 hour.
将反应液空气浴中减压浓缩,得到1.0克黄色固体1-氧杂-8-氮杂螺[4.5]癸-3-醇的三氟乙酸盐粗品(收率：94.8％)。The reaction solution was concentrated under reduced pressure in an air bath to obtain 1.0 g of crude trifluoroacetate of yellow solid 1-oxa-8-azaspiro[4.5]dec-3-ol (yield: 94.8%).
步骤B：合成5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step B: Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionylamino)-1H-indole-1,2 -Di-tert-butyl dicarboxylate
Figure PCTCN2020130361-appb-000080
Figure PCTCN2020130361-appb-000080
室温下,将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(178毫克，0.2毫摩尔)和1-氧杂-8-氮杂螺[4.5]癸-3-醇的三氟乙酸盐(542毫克，2.0毫摩尔)溶于四氢呋喃(6.0毫升)中。随后,向上述溶液中N,N-二异丙基乙胺(0.7毫升,4毫摩尔)。在60℃反应12小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (178 mg, 0.2 mmol) and 1 The trifluoroacetate salt of -oxa-8-azaspiro[4.5]dec-3-ol (542 mg, 2.0 mmol) was dissolved in tetrahydrofuran (6.0 mL). Subsequently, add N,N-diisopropylethylamine (0.7 mL, 4 mmol) to the above solution. React at 60°C for 12 hours.
向反应液中加水淬灭,混合液用乙酸乙酯(30毫升×3次)萃取,合并有机相,有机相用饱和食盐水(20毫升×3次),无水硫酸钠干燥,减压浓缩得到190毫克黄色固体5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：100％)。LCMS：RT＝4.14min,[M-H] -＝951.26。 The reaction solution was quenched by adding water, the mixed solution was extracted with ethyl acetate (30 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (20 ml × 3 times), dried with anhydrous sodium sulfate, and concentrated under reduced pressure Obtain 190 mg of yellow solid 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(3-hydroxy-1-oxa-8-azaspiro[4.5]decane-8-formylamino)phenyl)propionylamino)-1H-indole-1, Di-tert-butyl 2-dicarboxylic acid (yield: 100%). LCMS: RT = 4.14 min, [MH] - =951.26.
步骤C：合成5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step C: Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxy acid
Figure PCTCN2020130361-appb-000081
Figure PCTCN2020130361-appb-000081
将5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(190毫克,0.20毫摩尔)溶于二氯甲烷(5.0毫升)中。随后,向上述溶液中加入三氟乙酸(1.0毫升),在室温下搅拌1小时。Put 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxy Di-tert-butyl ester (190 mg, 0.20 mmol) was dissolved in dichloromethane (5.0 mL). Subsequently, trifluoroacetic acid (1.0 ml) was added to the above solution and stirred at room temperature for 1 hour.
将反应液空气浴中减压浓缩。将所得残余物用制备型高效液相色谱纯化,得到26毫克黄色固体5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：17.2％)。LCMS：RT＝3.24min,[M-H] -＝787.13。 1H NMR(500MHz,DMSO)δ12.89(s,1H),11.70(s,1H),10.12(s,1H),9.78(s,1H),8.43(s,1H),8.08–7.87(m,2H),7.78(dt,J＝8.6,5.3Hz,2H),7.49–7.25(m,3H),7.14(d,J＝7.9Hz,2H),7.06(s,1H),5.30(dd,J＝9.7,5.7Hz,1H),4.86(s,1H),4.32(s,1H),3.81(dd,J＝9.2,4.9Hz,1H),3.73(s,1H),3.61–3.51(m,3H),3.38(s,1H),3.23(d,J＝14.8Hz,1H),3.05–2.96(m,1H),1.87(dd,J＝13.0,6.6Hz,1H),1.68(ddd,J＝29.9,21.5,10.9Hz,3H),1.49(s,2H)。 The reaction solution was concentrated under reduced pressure in an air bath. The resulting residue was purified by preparative high performance liquid chromatography to obtain 26 mg of yellow solid 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl) -2,3-dioxopiperazin-1-yl)-3-(4-(3-hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl ) Propionamido)-1H-indole-2-carboxylic acid (yield: 17.2%). LCMS: RT = 3.24 min, [MH] - =787.13. 1 H NMR (500MHz, DMSO) δ 12.89 (s, 1H), 11.70 (s, 1H), 10.12 (s, 1H), 9.78 (s, 1H), 8.43 (s, 1H), 8.08-7.87 (m ,2H),7.78(dt,J=8.6,5.3Hz,2H),7.49–7.25(m,3H),7.14(d,J=7.9Hz,2H),7.06(s,1H),5.30(dd, J = 9.7, 5.7 Hz, 1H), 4.86 (s, 1H), 4.32 (s, 1H), 3.81 (dd, J = 9.2, 4.9 Hz, 1H), 3.73 (s, 1H), 3.61-3.51 (m ,3H), 3.38(s,1H), 3.23(d,J=14.8Hz,1H),3.05-2.96(m,1H),1.87(dd,J=13.0,6.6Hz,1H),1.68(ddd, J=29.9, 21.5, 10.9 Hz, 3H), 1.49 (s, 2H).
实施例15Example 15
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲基磺酰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(Methylsulfonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000082
Figure PCTCN2020130361-appb-000082
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲基磺酰基)哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(Methylsulfonyl)piperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000083
Figure PCTCN2020130361-appb-000083
室温下,将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(178毫克,0.2毫摩尔)和4-(甲基磺酰基)哌啶(163毫克,1.0毫摩尔)溶于四氢呋喃(6.0毫升)中。随后,向上述溶液中N,N-二异丙基乙胺(0.3毫升,2毫摩尔)。在60℃反应12小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (178 mg, 0.2 mmol) and 4 -(Methylsulfonyl)piperidine (163 mg, 1.0 mmol) was dissolved in tetrahydrofuran (6.0 mL). Subsequently, add N,N-diisopropylethylamine (0.3 mL, 2 mmol) to the above solution. React at 60°C for 12 hours.
向反应液中加水淬灭,混合液用乙酸乙酯(30毫升×3次)萃取,合并有机相,有机相用饱和食盐水(20毫升×3次),无水硫酸钠干燥,减压浓缩得到192毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲基磺酰基)哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯粗品(收率：100％)。LCMS：RT＝4.16min,[M-H] - ＝957.29。 The reaction solution was quenched by adding water, the mixed solution was extracted with ethyl acetate (30 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (20 ml × 3 times), dried with anhydrous sodium sulfate, and concentrated under reduced pressure Obtain 192 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester Crude product (yield: 100%). LCMS: RT = 4.16 min, [MH] - =957.29.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲基磺酰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000084
Figure PCTCN2020130361-appb-000084
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲基磺酰基)哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(192毫克,0.20毫摩尔)溶于二氯甲烷(5.0毫升)中。随后,向上述溶液中加入三氟乙酸(1.0毫升),在室温下搅拌1小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (192 mg, 0.20 mmol) was dissolved in dichloromethane (5.0 mL). Subsequently, trifluoroacetic acid (1.0 ml) was added to the above solution and stirred at room temperature for 1 hour.
将反应液空气浴中减压浓缩。将所得残余物用制备型高效液相色谱纯化,得到25毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲基磺酰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：15.0％)。LCMS：RT＝3.24min,[M-H] -＝801.18。 1H NMR(400MHz,DMSO)δ13.08–12.69(m,1H),11.71(s,1H),10.13(s,1H),9.78(s,1H),8.55(s,1H),8.00(s,1H),7.95(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.3Hz,1H),7.39(dd,J＝11.2,8.7Hz,3H),7.33(dd,J＝8.9,1.9Hz,1H),7.16(d,J＝8.5Hz,2H),7.06(d,J＝1.5Hz,1H),5.30(dd,J＝9.6,5.9Hz,1H),4.26(d,J＝13.4Hz,2H),3.74(s,4H),3.29–3.23(m,1H),3.22(s,1H),3.03(d,J＝9.8Hz,1H),2.94(s,3H),2.83(t,J＝12.0Hz,2H),2.03(d,J＝8.5Hz,2H),1.53(d,J＝11.9Hz,2H)。 The reaction solution was concentrated under reduced pressure in an air bath. The resulting residue was purified by preparative high performance liquid chromatography to obtain 25 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl) -2,3-dioxopiperazin-1-yl)-3-(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indyl Dole-2-carboxylic acid (yield: 15.0%). LCMS: RT = 3.24 min, [MH] - = 801.18. 1 H NMR (400MHz, DMSO) δ 13.08-12.69 (m, 1H), 11.71 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.55 (s, 1H), 8.00 (s , 1H), 7.95 (d, J = 2.2 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.76 (dd, J = 8.6, 2.3 Hz, 1H), 7.39 (dd, J = 11.2, 8.7Hz, 3H), 7.33 (dd, J = 8.9, 1.9 Hz, 1H), 7.16 (d, J = 8.5 Hz, 2H), 7.06 (d, J = 1.5 Hz, 1H), 5.30 (dd, J = 9.6, 5.9 Hz, 1H), 4.26 (d, J = 13.4 Hz, 2H), 3.74 (s, 4H), 3.29-3.23 (m, 1H), 3.22 (s, 1H), 3.03 (d, J = 9.8 Hz, 1H), 2.94 (s, 3H), 2.83 (t, J = 12.0 Hz, 2H), 2.03 (d, J = 8.5 Hz, 2H), 1.53 (d, J = 11.9 Hz, 2H).
实施例16Example 16
合成(S)-5-(3-(4-(4-羧基哌啶-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(3-(4-(4-carboxypiperidine-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H-tetrazole-1- (Yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000085
Figure PCTCN2020130361-appb-000085
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(3-(4-(4-(叔丁氧基羰基)哌啶-1-甲酰胺基)苯基)-2-(4-(5-氯-2-(1H-四唑)-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(3-(4-(4-(tert-butoxycarbonyl)piperidine-1-carboxamido)phenyl)-2-(4-(5-chloro-2 -(1H-tetrazole)-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000086
Figure PCTCN2020130361-appb-000086
室温下,将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(178毫克,0.2毫摩尔)和哌啶-4-羧酸叔丁酯(251毫克,1.0毫摩尔)溶于四氢呋喃(6.0毫升)中。随后,向上述溶液中N,N-二异丙基乙胺(0.3毫升,2毫摩尔)。在60℃反应12小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (178 mg, 0.2 mmol) and piperidine Tert-butyl pyridine-4-carboxylate (251 mg, 1.0 mmol) was dissolved in tetrahydrofuran (6.0 mL). Subsequently, add N,N-diisopropylethylamine (0.3 mL, 2 mmol) to the above solution. React at 60°C for 12 hours.
向反应液中加水淬灭,混合液用乙酸乙酯(30毫升×3次)萃取,合并有机相,有机相用饱和食盐水(20毫升×3次),无水硫酸钠干燥,减压浓缩得到76毫克黄色固体(S)-5-(3-(4-(4-(叔丁氧基羰基)哌啶-1-甲酰胺基)苯基)-2-(4-(5-氯-2-(1H-四唑)-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯粗品(收率：100％)。LCMS：RT＝4.57min,[M-H] -＝979.36。 The reaction solution was quenched by adding water, the mixed solution was extracted with ethyl acetate (30 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (20 ml × 3 times), dried with anhydrous sodium sulfate, and concentrated under reduced pressure To obtain 76 mg of yellow solid (S)-5-(3-(4-(4-(tert-butoxycarbonyl)piperidine-1-carboxamido)phenyl)-2-(4-(5-chloro- 2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionylamino)-1H-indole-1,2-dicarboxylic acid Crude butyl ester (yield: 100%). LCMS: RT = 4.57 min, [MH] - =979.36.
步骤B：合成(S)-5-(3-(4-(4-羧基哌啶-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(3-(4-(4-carboxypiperidine-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H-tetrazole) -1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000087
Figure PCTCN2020130361-appb-000087
将(S)-5-(3-(4-(4-(叔丁氧基羰基)哌啶-1-甲酰胺基)苯基)-2-(4-(5-氯-2-(1H-四唑)-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(76毫克,0.20毫摩尔)溶于二氯甲烷(5.0毫升)中。随后,向上述溶液中加入三氟乙酸(1.0毫升),在室温下搅拌1小时。Add (S)-5-(3-(4-(4-(tert-butoxycarbonyl)piperidine-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H -Tetrazol)-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (76 Mg, 0.20 mmol) was dissolved in dichloromethane (5.0 mL). Subsequently, trifluoroacetic acid (1.0 ml) was added to the above solution and stirred at room temperature for 1 hour.
将反应液空气浴中减压浓缩。将所得残余物用制备型高效液相色谱纯化,得到4毫克黄色固体(S)-5-(3-(4-(4-羧基哌啶-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸(收率：6.4％)。LCMS：RT＝3.24min,[M-H] -＝769.18。 1H NMR(500MHz,DMSO)δ12.81(s,2H),11.71(s,1H),10.12(s,1H),9.78(s,1H),8.45(d,J＝4.6Hz,1H),8.06–7.90(m,2H),7.82(d,J＝8.6Hz,1H),7.76(dd,J＝8.5,2.2Hz,1H),7.39(dd,J＝13.7,8.7Hz,3H),7.33(d,J＝8.8Hz,1H),7.14(d,J＝7.9Hz,2H),7.06(s,1H),5.30(dd,J＝9.8,5.8Hz,1H),4.01(d,J＝13.3Hz,2H),3.74(s,2H),3.62(s,1H),3.24(s,2H),3.17(s,1H),3.08–2.96(m,1H),2.90(t,J＝12.2Hz,2H),1.82(d,J＝9.7Hz,2H),1.57–1.40(m,2H)。 The reaction solution was concentrated under reduced pressure in an air bath. The resulting residue was purified by preparative high performance liquid chromatography to obtain 4 mg of yellow solid (S)-5-(3-(4-(4-carboxypiperidine-1-carboxamido)phenyl)-2-( 4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxy Acid (yield: 6.4%). LCMS: RT = 3.24 min, [MH] - =769.18. 1 H NMR (500MHz, DMSO) δ 12.81 (s, 2H), 11.71 (s, 1H), 10.12 (s, 1H), 9.78 (s, 1H), 8.45 (d, J = 4.6 Hz, 1H), 8.06–7.90 (m, 2H), 7.82 (d, J = 8.6 Hz, 1H), 7.76 (dd, J = 8.5, 2.2 Hz, 1H), 7.39 (dd, J = 13.7, 8.7 Hz, 3H), 7.33 (d,J=8.8Hz,1H), 7.14(d,J=7.9Hz,2H), 7.06(s,1H), 5.30(dd,J=9.8,5.8Hz,1H), 4.01(d,J= 13.3Hz, 2H), 3.74 (s, 2H), 3.62 (s, 1H), 3.24 (s, 2H), 3.17 (s, 1H), 3.08-2.96 (m, 1H), 2.90 (t, J = 12.2 Hz, 2H), 1.82 (d, J = 9.7 Hz, 2H), 1.57-1.40 (m, 2H).
实施例17Example 17
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(甲基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-(methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000088
Figure PCTCN2020130361-appb-000088
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(甲基磺酰基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(Methylsulfonyl)ureido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000089
Figure PCTCN2020130361-appb-000089
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(178毫克，0.20毫摩尔)和甲基磺酰胺(190毫克，2.0毫摩尔)的四氢呋喃(6.0毫 升)中滴加N,N-二异丙基乙胺(516毫克，4.0毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (178 mg, 0.20 mmol) N,N-diisopropylethylamine (516 mg, 4.0 mmol) was added dropwise to methylsulfonamide (190 mg, 2.0 mmol) in tetrahydrofuran (6.0 ml), and the dripping was completed, and the reaction was carried out at 60 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝10/1)。得到100毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(甲基磺酰基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：56.0％)。LCMS:RT＝4.20min,[M+H] +＝891.23。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1). Obtain 100 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(3-(methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (Yield: 56.0% ). LCMS: RT = 4.20 min, [M+H] + =891.23.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(甲基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000090
Figure PCTCN2020130361-appb-000090
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(甲基磺酰基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(100毫克，0.11毫摩尔)加入二氯甲烷(4.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到56毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(甲基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：60.8％)。LCMS:RT＝3.27min,[M-H] -＝733.08。 At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(Methylsulfonyl)ureido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (100 mg, 0.11 mmol ) Was added to methylene chloride (4.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours. When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 56 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiprazin-1-yl)-3-(4-(3-(methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 60.8 %). LCMS: RT = 3.27 min, [MH] - =733.08.
实施例18Example 18
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(乙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000091
Figure PCTCN2020130361-appb-000091
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(乙基磺酰基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000092
Figure PCTCN2020130361-appb-000092
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(178毫克，0.20毫摩尔)和乙基磺酰胺(436毫克，4.0毫摩尔)的四氢呋喃(6.0毫升)中滴加N,N-二异丙基乙胺(775毫克，6.0毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (178 mg, 0.20 mmol) N,N-diisopropylethylamine (775 mg, 6.0 mmol) was added dropwise to ethylsulfonamide (436 mg, 4.0 mmol) in tetrahydrofuran (6.0 mL), and the dripping was completed, and the reaction was carried out at 60 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝10/1)。得到110毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(乙基磺酰基)脲基)苯基)丙酰 氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：60.7％)。LCMS:RT＝4.24min,[M+Na] +＝927.14。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1). Obtain 110 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (Yield: 60.7% ). LCMS: RT = 4.24 min, [M+Na] + =927.14.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(乙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000093
Figure PCTCN2020130361-appb-000093
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(乙基磺酰基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(110毫克，0.12毫摩尔)加入二氯甲烷(4.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到48毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(乙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：53.3％)。LCMS:RT＝3.33min,[M+H] +＝749.03。 1H NMR(400MHz,DMSO)δ12.91(s,1H),11.72(s,1H),10.19(s,1H),10.13(s,1H),9.78(s,1H),8.79(s,1H),8.00(s,1H),7.95(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.3Hz,1H),7.38(d,J＝8.6Hz,3H),7.36–7.30(m,1H),7.22(d,J＝7.8Hz,2H),7.06(d,J＝1.5Hz,1H),5.36–5.26(m,1H),4.25–3.55(m,4H),3.44(dd,J＝14.6,7.2Hz,2H),3.25(dd,J＝14.2,5.2Hz,1H),3.12–3.00(m,1H),1.26(t,J＝7.4Hz,3H)。 At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (110 mg, 0.12 mmol ) Was added to methylene chloride (4.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours. When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 48 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 53.3 %). LCMS: RT = 3.33 min, [M+H] + =749.03. 1 H NMR (400MHz, DMSO) δ 12.91 (s, 1H), 11.72 (s, 1H), 10.19 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.79 (s, 1H) ),8.00(s,1H),7.95(d,J=2.2Hz,1H),7.82(d,J=8.6Hz,1H),7.76(dd,J=8.6,2.3Hz,1H),7.38(d ,J=8.6Hz,3H),7.36-7.30(m,1H),7.22(d,J=7.8Hz,2H),7.06(d,J=1.5Hz,1H),5.36-5.26(m,1H) ,4.25-3.55(m,4H),3.44(dd,J=14.6,7.2Hz,2H), 3.25(dd,J=14.2,5.2Hz,1H), 3.12-3.00(m,1H), 1.26(t , J=7.4Hz, 3H).
实施例19Example 19
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-((4-((S)-3-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -((4-((S)-3-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000094
Figure PCTCN2020130361-appb-000094
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-3-羟基哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-((S)-3-hydroxypiperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000095
Figure PCTCN2020130361-appb-000095
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(178毫克，0.20毫摩尔)和(S)-哌啶-3-醇盐酸盐(550毫克，4.0毫摩尔)的四氢呋喃(6.0毫升)中滴加N,N-二异丙基乙胺(1.03克，8.0毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (178 mg, 0.20 mmol) And (S)-piperidin-3-ol hydrochloride (550 mg, 4.0 mmol) in tetrahydrofuran (6.0 ml) was added dropwise N,N-diisopropylethylamine (1.03 g, 8.0 mmol), After dripping, react overnight at 60 degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝10/1)。得到93毫克白色固体(5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-3-羟基哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：52.0％)。LCMS:RT＝4.16min,[M+H] +＝867.22。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1). Obtain 93 mg of white solid (5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1 -Yl)-3-(4-((S)-3-hydroxypiperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (Yield: 52.0%). LCMS: RT = 4.16 min, [M+H] + =867.22.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-((4-((S)-3-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-((4-((S)-3-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000096
Figure PCTCN2020130361-appb-000096
室温下，将(5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-3-羟基哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(93毫克，0.10毫摩尔)加入二氯甲烷(4.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到48毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-((4-((S)-3-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：64.8％)。LCMS:RT＝3.20min,[M-H] -＝739.16。δ11.73(s,1H),10.14(s,1H),9.79(s,1H),8.42(s,1H),8.01(s,1H),7.95(d,J＝2.3Hz,1H),7.82(d,J＝8.5Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.41(s,1H),7.36(s,1H),7.32(dd,J＝8.8,1.9Hz,1H),7.14(d,J＝8.3Hz,2H),7.06(d,J＝2.1Hz,1H),5.33–5.26(m,1H),3.88–3.72(m,7H),3.48–3.40(m,1H),3.26–3.18(m,1H),3.05–2.96(m,1H),2.92–2.83(m,1H),2.72–2.65(m,1H),2.03–1.95(m,1H),1.89–1.80(m,1H),1.72–1.62(m,1H),1.38–1.30(m,1H)。 At room temperature, the (5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((S)-3-hydroxypiperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester ( 93 mg, 0.10 mmol) was added to dichloromethane (4.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours. After the reaction was over, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The material was dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid precipitated out. The filter cake was filtered with suction. The filter cake was washed with n-hexane and dried to obtain 48 mg of white solid 5-((S )-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-((4-( (S)-3-Hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 64.8%). LCMS: RT = 3.20 min, [MH ] - ＝739.16. δ11.73(s,1H),10.14(s,1H),9.79(s,1H),8.42(s,1H),8.01(s,1H),7.95(d,J=2.3Hz ,1H),7.82(d,J=8.5Hz,1H),7.77(dd,J=8.6,2.3Hz,1H),7.41(s,1H),7.36(s,1H),7.32(dd,J= 8.8,1.9Hz,1H), 7.14(d,J=8.3Hz,2H), 7.06(d,J=2.1Hz,1H), 5.33–5.26(m,1H), 3.88–3.72(m,7H), 3.48–3.40(m,1H), 3.26–3.18(m,1H), 3.05–2.96(m,1H), 2.92–2.83(m,1H), 2.72–2.65(m,1H), 2.03–1.95(m ,1H), 1.89–1.80(m,1H), 1.72–1.62(m,1H), 1.38–1.30(m,1H).
实施例20Example 20
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基)环丁基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(3-(hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000097
Figure PCTCN2020130361-appb-000097
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成3-(3-氧代哌嗪-1-基)环丁烷-1-羧酸甲酯Step A: Synthesis of methyl 3-(3-oxopiperazin-1-yl)cyclobutane-1-carboxylate
Figure PCTCN2020130361-appb-000098
Figure PCTCN2020130361-appb-000098
室温下，向含有哌嗪-2-酮(1.00克，10.0毫摩尔)和3-氧代环丁烷-1-羧酸甲酯(1.92克，15.0毫摩尔)的甲醇(20.0毫升)溶液中加入氰基硼氢化钠(8.7毫克，13.0毫摩尔)和乙酸(0.4毫升)，加毕，室温反应2小时。To a solution of piperazin-2-one (1.00 g, 10.0 mmol) and methyl 3-oxocyclobutane-1-carboxylate (1.92 g, 15.0 mmol) in methanol (20.0 mL) at room temperature Sodium cyanoborohydride (8.7 mg, 13.0 mmol) and acetic acid (0.4 ml) were added, after the addition, the reaction was carried out at room temperature for 2 hours.
反应结束，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝10/1)。得到954毫克淡黄色油状物3-(3-氧代哌嗪-1-基)环丁烷-1-羧酸甲酯(收率45.0％)。LCMS:RT＝0.69min,[M+H] +＝213.11。 After the reaction was completed, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1). 954 mg of methyl 3-(3-oxopiperazin-1-yl)cyclobutane-1-carboxylate was obtained as a pale yellow oil (yield 45.0%). LCMS: RT=0.69 min, [M+H] + =213.11.
步骤B：合成4-(3-(羟甲基)环丁基)哌嗪-2-酮Step B: Synthesis of 4-(3-(hydroxymethyl)cyclobutyl)piperazin-2-one
Figure PCTCN2020130361-appb-000099
Figure PCTCN2020130361-appb-000099
室温下，向含有3-(3-氧代哌嗪-1-基)环丁烷-1-羧酸甲酯(848毫克，4.0毫摩尔)的四氢呋喃/甲醇(20.0/20.0毫升)溶液中加入硼氢化锂(88毫克，4.0毫摩尔)，加毕，加热至40摄氏度反应3小时。At room temperature, add 3-(3-oxopiperazin-1-yl)cyclobutane-1-carboxylic acid methyl ester (848 mg, 4.0 mmol) in tetrahydrofuran/methanol (20.0/20.0 ml) solution Lithium borohydride (88 mg, 4.0 mmol), after the addition, heat to 40 degrees Celsius and react for 3 hours.
反应结束，加水(5毫升)淬灭，二氯甲烷萃取(30毫升×2次)，无水硫酸钠干燥，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝5/1)。得到412毫克淡黄色油状物4-(3-(羟甲基)环丁基)哌嗪-2-酮(56.0％)。LCMS:RT＝0.76min,[M+H] +＝185.09。 After the reaction was completed, it was quenched by adding water (5 ml), extracted with dichloromethane (30 ml × 2 times), dried over anhydrous sodium sulfate, and evaporated under reduced pressure to remove the solvent. The residue obtained was purified by silica gel column chromatography (eluent: two Methyl chloride/methanol = 5/1). 412 mg of 4-(3-(hydroxymethyl)cyclobutyl)piperazin-2-one (56.0%) was obtained as a pale yellow oil. LCMS: RT=0.76 min, [M+H] + =185.09.
步骤C：合成(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基))环丁基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(3-(Hydroxymethyl))cyclobutyl)-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000100
Figure PCTCN2020130361-appb-000100
氮气保护下，向含有(S)-3-(4-溴苯基)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(602毫克，1.0毫摩尔)、4-(3-(羟甲基)环丁基)哌嗪-2-酮(184毫克，1.0毫摩尔)的甲苯(5.0毫升)溶液中加入碘化亚铜(190毫克，1.0毫摩尔)，碳酸铯(656毫克，2.0毫摩尔)，N 1,N 2-二甲基乙二胺(176毫克，2.0毫摩尔)，加毕，加热至110摄氏度反应过夜。 Under the protection of nitrogen, to contain (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxo Piperazin-1-yl) tert-butyl propionate (602 mg, 1.0 mmol), 4-(3-(hydroxymethyl)cyclobutyl) piperazin-2-one (184 mg, 1.0 mmol) Toluene (5.0 ml) was added cuprous iodide (190 mg, 1.0 mmol), cesium carbonate (656 mg, 2.0 mmol), N 1 ,N 2 -dimethylethylenediamine (176 mg, 2.0 mmol) Mol), after the addition, heat to 110 degrees Celsius and react overnight.
反应结束，加水(5毫升)淬灭，乙酸乙酯萃取(40毫升×2次)，合并有机相，用饱和食盐水(50毫升)洗，无水硫酸钠干燥，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝30/1)。得到352毫克淡黄色油状物(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基))环丁基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(50.0％)。LCMS:RT＝3.30min,[M+H] +＝706.18。 After the reaction was completed, it was quenched with water (5 mL), extracted with ethyl acetate (40 mL×2 times), and the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=30/1). Obtain 352 mg of light yellow oil (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(3-(Hydroxymethyl))cyclobutyl)-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate (50.0%). LCMS: RT = 3.30 min, [M+H] + =706.18.
步骤D：合成(S)-2-(4-(2-氨基-5-氯-苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基))环丁基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-(4-(2-amino-5-chloro-phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-( 3-(Hydroxymethyl))cyclobutyl)-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000101
Figure PCTCN2020130361-appb-000101
氮气保护下，向含有(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基))环丁基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(352毫克，0.50毫摩尔)的二氯甲烷(4.0毫升)溶液中加入二甲基巴比妥酸(624毫克，4.0毫摩尔)，四三苯基膦钯(29毫克，0.025毫摩尔)，加毕，加热至40摄氏度反应过夜。Under the protection of nitrogen, it contains (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-(4-(3-(hydroxymethyl))cyclobutyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (352 mg, 0.50 mmol) in dichloride Add dimethylbarbituric acid (624 mg, 4.0 mmol) and tetrakistriphenylphosphine palladium (29 mg, 0.025 mmol) to the methane (4.0 mL) solution. After the addition, heat to 40 degrees Celsius and react overnight.
反应结束，用二氯甲烷(100毫升)稀释，依次用饱和碳酸氢钠(50毫升)、饱和食盐水(50毫升)洗涤，无水硫酸钠干燥，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝30/1)。得到220毫克淡黄色油状物 (S)-2-(4-(2-氨基-5-氯-苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基))环丁基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(70.4％)。LCMS:RT＝3.16min,[M+Na] +＝648.23。 After the reaction, it was diluted with dichloromethane (100 mL), washed with saturated sodium bicarbonate (50 mL), saturated brine (50 mL), dried over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure, and the resulting residue was used silica gel Purification by column chromatography (eluent: dichloromethane/methanol=30/1). Obtain 220 mg of light yellow oil (S)-2-(4-(2-amino-5-chloro-phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-( Tert-Butyl 4-(3-(hydroxymethyl))cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionate (70.4%). LCMS: RT = 3.16 min, [M+Na] + =648.23.
步骤E：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基)环丁基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁基酯Step E: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(3-(Hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000102
Figure PCTCN2020130361-appb-000102
氮气保护下，向含有((S)-2-(4-(2-氨基-5-氯-苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基))环丁基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(220毫克，0.35毫摩尔)乙酸(1.0毫升)溶液中加入原甲酸三乙酯(311毫克，2.1毫摩尔)，叠氮化钠(114毫克，1.75毫摩尔)，加毕，加热至80摄氏度反应2小时。Under the protection of nitrogen, to contain ((S)-2-(4-(2-amino-5-chloro-phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-( 4-(3-(Hydroxymethyl))cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionic acid tert-butyl ester (220 mg, 0.35 mmol) in acetic acid (1.0 ml) solution Add triethyl orthoformate (311 mg, 2.1 mmol), sodium azide (114 mg, 1.75 mmol), after the addition, heat to 80 degrees Celsius and react for 2 hours.
反应结束，加亚硝酸钠(200毫克)淬灭，乙酸乙酯萃取(40毫升×2次)，合并有机相，用饱和食盐水(50毫升)洗，无水硫酸钠干燥，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝20/1)。得到180毫克淡黄色油状物(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基)环丁基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁基酯(收率75.3％)。LCMS:RT＝2.91min,[M+H] +＝679.16。 After the reaction is over, add sodium nitrite (200 mg) to quench, extract with ethyl acetate (40 ml×2 times), combine the organic phases, wash with saturated brine (50 ml), dry with anhydrous sodium sulfate, and evaporate under reduced pressure Solvent, the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=20/1). Obtain 180 mg of light yellow oil (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(3-(hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (yield 75.3%). LCMS: RT = 2.91 min, [M+H] + =679.16.
步骤F：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-(-(3-(羟甲基)环丁基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step F: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-(-(3-(hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2 -Di-tert-butyl dicarboxylate
Figure PCTCN2020130361-appb-000103
Figure PCTCN2020130361-appb-000103
室温下，向含有(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基)环丁基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁基酯(100毫克，0.15毫摩尔)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(1.0毫升)，加毕，室温反应1.5小时。减压浓缩，所得粗产物直接用于下一步反应。At room temperature, it contains (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-butyl 3-(4-(4-(3-(hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionate (100 mg, 0.15 mmol) Trifluoroacetic acid (1.0 ml) was added to the dichloromethane (4.0 ml) solution. After the addition, the reaction was carried out at room temperature for 1.5 hours. It was concentrated under reduced pressure, and the obtained crude product was directly used in the next reaction.
将上述粗产物和5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯(133毫克，0.40毫摩尔)溶于无水N,N-二甲基甲酰胺(2.0毫升)，向该溶液中加入HATU(152毫克，0.40毫摩尔)，二异丙基乙基胺(77毫克，0.60毫摩尔)，室温反应2小时。反应结束，加水(10毫升)淬灭，乙酸乙酯萃取(40毫升×2次)，合并有机相，依次用水(40毫升×2次)以及饱和食盐水(50毫升)洗涤，无水硫酸钠干燥，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝30/1)。得到110毫克淡黄色油状物(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-(-(3-(羟甲基)环丁基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率78.2％)。LCMS:RT＝3.45min,[M+H] +＝937.28。 The above crude product and di-tert-butyl 5-amino-1H-indole-1,2-dicarboxylate (133 mg, 0.40 mmol) were dissolved in anhydrous N,N-dimethylformamide (2.0 mL) Add HATU (152 mg, 0.40 mmol) and diisopropylethylamine (77 mg, 0.60 mmol) to the solution, and react at room temperature for 2 hours. After the reaction is complete, add water (10 ml) to quench, extract with ethyl acetate (40 ml × 2 times), combine the organic phases, and wash with water (40 ml × 2 times) and saturated brine (50 ml), anhydrous sodium sulfate After drying, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=30/1). Obtain 110 mg of light yellow oil (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-(-4-(-(3-(hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole- Di-tert-butyl 1,2-dicarboxylate (yield 78.2%). LCMS: RT = 3.45 min, [M+H] + =937.28.
步骤G：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基)环丁基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step G: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(3-(hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxy acid
Figure PCTCN2020130361-appb-000104
Figure PCTCN2020130361-appb-000104
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-(-(3-(羟甲基)环丁基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(110毫克，0.12毫摩尔)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(1.0毫升)，加毕，室温反应1小时。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(-4-(-(3-(hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-1, Trifluoroacetic acid (1.0 mL) was added to a di-tert-butyl 2-dicarboxylate (110 mg, 0.12 mmol) solution in dichloromethane (4.0 mL). After the addition, the reaction was carried out at room temperature for 1 hour.
减压浓缩，所得粗产物溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到78毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(3-(羟甲基)环丁基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸三氟乙酸盐。(收率74.5％)。LCMS:RT＝2.69min,[M+H] +＝781.08。 1H NMR(400MHz,DMSO)δ11.75(s,1H),10.15(s,1H),9.77(s,1H),8.00(s,1H),7.94(d,J＝2.2Hz,1H),7.81(d,J＝8.5Hz,1H),7.77(dd,J＝8.6,2.2Hz,1H),7.41–7.29(m,5H),7.07(d,J＝1.5Hz,1H),5.43–5.30(m,1H),3.70–3.58(m,13H),3.38(d,J＝5.1Hz,2H),2.39–2.25(m,2H),2.19–2.08(m,1H),2.04–1.91(m,2H)。 Concentrated under reduced pressure, the obtained crude product was dissolved in dichloromethane (1.0 ml), and added dropwise to n-hexane (10.0 ml). A white solid precipitated out. The filter cake was washed with n-hexane and dried to obtain 78 mg of white solid. (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-(4-(3-(Hydroxymethyl)cyclobutyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid trifluoroethane Acid salt. (Yield 74.5%). LCMS: RT = 2.69 min, [M+H] + =781.08. 1 H NMR (400MHz, DMSO) δ 11.75 (s, 1H), 10.15 (s, 1H), 9.77 (s, 1H), 8.00 (s, 1H), 7.94 (d, J = 2.2 Hz, 1H), 7.81(d,J=8.5Hz,1H),7.77(dd,J=8.6,2.2Hz,1H),7.41-7.29(m,5H),7.07(d,J=1.5Hz,1H),5.43-5.30 (m,1H),3.70–3.58(m,13H), 3.38(d,J=5.1Hz,2H), 2.39–2.25(m,2H), 2.19–2.08(m,1H),2.04–1.91(m ,2H).
实施例21Example 21
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000105
Figure PCTCN2020130361-appb-000105
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(四氢呋喃-3-基)甲基4-甲基苯磺酸酯Step A: Synthesis of (tetrahydrofuran-3-yl)methyl 4-methylbenzenesulfonate
Figure PCTCN2020130361-appb-000106
Figure PCTCN2020130361-appb-000106
室温下，向含有(四氢呋喃-3-基)甲醇(1.00克，10.0毫摩尔)和三乙胺(3.04克，30.0毫摩尔)的二氯甲烷(20.0毫升)溶液中加入对甲基苯磺酰氯(2.86克，15.0毫摩尔)，加毕，室温反应5小时。At room temperature, add p-toluenesulfonyl chloride to a dichloromethane (20.0 ml) solution containing (tetrahydrofuran-3-yl)methanol (1.00 g, 10.0 mmol) and triethylamine (3.04 g, 30.0 mmol) (2.86 g, 15.0 mmol), after the addition, react at room temperature for 5 hours.
反应结束，用二氯甲烷(150毫升)稀释，依次用水(80毫升)、饱和食盐水(80毫升)洗涤，无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：正己烷/乙酸乙酯＝4/1)。得到2.1克淡黄色油状物(四氢呋喃-3-基)甲基4-甲基苯磺酸酯(收率81.9％)。LCMS:RT＝3.72min,[M+H] +＝257.07。 After the reaction was completed, it was diluted with dichloromethane (150 mL), washed with water (80 mL), saturated brine (80 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (washing Removal agent: n-hexane/ethyl acetate=4/1). 2.1 g of light yellow oily (tetrahydrofuran-3-yl)methyl 4-methylbenzenesulfonate was obtained (yield 81.9%). LCMS: RT = 3.72 min, [M+H] + =257.07.
步骤B：合成4-((四氢呋喃-3-基)甲基)哌嗪-2-酮Step B: Synthesis of 4-((tetrahydrofuran-3-yl)methyl)piperazin-2-one
Figure PCTCN2020130361-appb-000107
Figure PCTCN2020130361-appb-000107
室温下，向含有哌嗪-2-酮(2.1克，21.0毫摩尔)和(四氢呋喃-3-基)甲基4-甲基苯磺酸酯(3.7克，14.0毫摩尔)的乙腈(50.0毫升)溶液中加入碳酸钾(3.9克，28.0毫摩尔)和碘化钠(105毫克，0.7毫摩尔)，加毕，加热至80摄氏度反应6小时。At room temperature, to acetonitrile (50.0 ml) containing piperazin-2-one (2.1 g, 21.0 mmol) and (tetrahydrofuran-3-yl) methyl 4-methylbenzenesulfonate (3.7 g, 14.0 mmol) ) Potassium carbonate (3.9 g, 28.0 mmol) and sodium iodide (105 mg, 0.7 mmol) were added to the solution. After the addition, the solution was heated to 80 degrees Celsius and reacted for 6 hours.
反应结束，过滤，滤液减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝10/1)。得到1.7克淡黄色油状物3-(3-氧代哌嗪-1-基)环丁烷-1-羧酸甲酯(收率66.7％)。LCMS:RT＝0.67min,[M+H] +＝185.13。 After the reaction was completed, the filtrate was filtered and the filtrate was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=10/1). 1.7 g of methyl 3-(3-oxopiperazin-1-yl)cyclobutane-1-carboxylate was obtained as a pale yellow oil (yield 66.7%). LCMS: RT=0.67 min, [M+H] + =185.13.
步骤C：合成(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000108
Figure PCTCN2020130361-appb-000108
氮气保护下，向含有(S)-3-(4-溴苯基)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(542毫克，0.90毫摩尔)、3-(3-氧代哌嗪-1-基)环丁烷-1-羧酸甲酯(331毫克，1.8毫摩尔)的甲苯(4.5毫升)溶液中加入碘化亚铜(171毫克，0.90毫摩尔)，碳酸铯(590毫克，1.8毫摩尔)，N 1,N 2-二甲基乙二胺(158毫克，1.8毫摩尔)，加毕，加热至110摄氏度反应过夜。 Under the protection of nitrogen, to contain (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxo Piperazin-1-yl) tert-butyl propionate (542 mg, 0.90 mmol), methyl 3-(3-oxopiperazin-1-yl)cyclobutane-1-carboxylate (331 mg, 1.8 MM) in toluene (4.5 mL) was added cuprous iodide (171 mg, 0.90 mmol), cesium carbonate (590 mg, 1.8 mmol), N 1 ,N 2 -dimethylethylenediamine (158 Mg, 1.8 mmol), after the addition, heat to 110 degrees Celsius and react overnight.
反应结束，加水(5毫升)淬灭，乙酸乙酯萃取(40毫升×2次)，合并有机相，用饱和食盐水(50毫升)洗，无水硫酸钠干燥，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝30/1)。得到400毫克淡黄色油状物(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(62.9％)。LCMS:RT＝3.67min,[M+H] +＝706.06。 After the reaction was completed, it was quenched with water (5 mL), extracted with ethyl acetate (40 mL×2 times), and the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=30/1). Obtain 400 mg of light yellow oil (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (62.9%). LCMS: RT = 3.67 min, [M+H] + = 706.06.
步骤D：合成(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(tetrahydrofuran) -3-yl)methyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000109
Figure PCTCN2020130361-appb-000109
氮气保护下，向含有(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(400毫克，0.56毫摩尔)的二氯甲烷(4.0毫升)溶液中加入二甲基巴比妥酸(700毫克，4.48毫摩尔)，四三苯基膦钯(32毫克，0.027毫摩尔)，加毕，加热至40摄氏度反应过夜。Under the protection of nitrogen, it contains (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-(4-(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (400 mg, 0.56 mmol) in dichloromethane (4.0 mL ) Add dimethylbarbituric acid (700 mg, 4.48 mmol) and tetrakistriphenylphosphine palladium (32 mg, 0.027 mmol) to the solution. After the addition, heat to 40 degrees Celsius and react overnight.
反应结束，用二氯甲烷(100毫升)稀释，依次用饱和碳酸氢钠(50毫升)、饱和食盐水(50毫升)洗涤，无水硫酸钠干燥，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝30/1)。得到300毫克淡黄色油状物(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(85.6％)。LCMS:RT＝3.08min,[M+H] +＝626.58。 After the reaction, it was diluted with dichloromethane (100 mL), washed with saturated sodium bicarbonate (50 mL), saturated brine (50 mL), dried over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure, and the resulting residue was used silica gel Purification by column chromatography (eluent: dichloromethane/methanol=30/1). Obtain 300 mg of light yellow oil (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4 -(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate (85.6%). LCMS: RT=3.08min, [M+H] + =626.58.
步骤E：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酸叔丁基酯Step E: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000110
Figure PCTCN2020130361-appb-000110
氮气保护下，向含有(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(300毫克，0.48毫摩尔)乙酸(1.0毫升)溶液中加入原甲酸三乙酯(427毫克，2.88毫摩尔)，叠氮化钠(156毫克，2.4毫摩尔)，加毕，加热至80摄氏度反应2小时。Under the protection of nitrogen, to contain (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4- (Tetrahydrofuran-3-yl) methyl-2-oxopiperazin-1-yl) phenyl) tert-butyl propionate (300 mg, 0.48 mmol) in acetic acid (1.0 mL) was added triethyl orthoformate (427 mg, 2.88 mmol), sodium azide (156 mg, 2.4 mmol), after the addition, heat to 80 degrees Celsius and react for 2 hours.
反应结束，加亚硝酸钠(200毫克)淬灭，乙酸乙酯萃取(40毫升×2次)，合并有机相，用饱和食盐水(50毫升)洗，无水硫酸钠干燥，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝20/1)。得到200毫克淡黄色油状物(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酸叔丁基酯(收率61.4％)。LCMS:RT＝3.03min,[M+H] +＝679.32。 After the reaction is over, add sodium nitrite (200 mg) to quench, extract with ethyl acetate (40 ml×2 times), combine the organic phases, wash with saturated brine (50 ml), dry with anhydrous sodium sulfate, and evaporate under reduced pressure Solvent, the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=20/1). Obtain 200 mg of light yellow oil (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate (yield 61.4%). LCMS: RT=3.03min, [M+H] + =679.32.
步骤F：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step F: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester
Figure PCTCN2020130361-appb-000111
Figure PCTCN2020130361-appb-000111
室温下，向含有(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酸叔丁基酯(200毫克，0.29毫摩尔)的二氯甲烷(8.0毫升)溶液中加入三氟乙酸(2.0毫升)，加毕，室温反应2小时。减压浓缩，所得粗产物直接用于下一步反应。LCMS:RT＝1.57min,[M-H] -＝621.11。 At room temperature, it contains (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-Butyl 3-(4-(4-(tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)propionate (200 mg, 0.29 mmol) in dichloromethane (8.0 mL) Trifluoroacetic acid (2.0 mL) was added to the solution. After the addition, the reaction was carried out at room temperature for 2 hours. It was concentrated under reduced pressure, and the obtained crude product was directly used in the next reaction. LCMS: RT=1.57 min, [MH] - =621.11.
将上述粗产物和5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯(199毫克，0.60毫摩尔)溶于无水N,N-二甲基甲酰胺(3.0毫升)，向该溶液中加入HATU(228毫克，0.60毫摩尔)，二异丙基乙基胺(116毫克，0.90毫摩尔)，室温反应2小时。The above crude product and di-tert-butyl 5-amino-1H-indole-1,2-dicarboxylate (199 mg, 0.60 mmol) were dissolved in anhydrous N,N-dimethylformamide (3.0 mL) Add HATU (228 mg, 0.60 mmol) and diisopropylethylamine (116 mg, 0.90 mmol) to the solution, and react at room temperature for 2 hours.
反应结束，加水(10毫克)淬灭，乙酸乙酯萃取(40毫升×2次)，合并有机相，依次用水(40毫升×2次)以及饱和食盐水(50毫升)洗涤，无水硫酸钠干燥，减压蒸除溶剂，所得残余物用硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝50/1)。得到153毫克淡黄色油状物(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率55.4％)。LCMS:RT＝3.76min,[M+H] +＝937.50。 After the reaction is over, add water (10 mg) to quench, extract with ethyl acetate (40 ml × 2 times), combine the organic phases, and wash with water (40 ml × 2 times) and saturated brine (50 ml), anhydrous sodium sulfate After drying, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol=50/1). 153 mg of light yellow oil (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-(4-((tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate (yield 55.4%). LCMS: RT = 3.76 min, [M+H] + =937.50.
步骤G：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step G: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000112
Figure PCTCN2020130361-appb-000112
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(153毫克，0.16毫摩尔)的二氯甲烷(4.0毫升)溶液中加入三氟乙酸(1.0毫升)，加毕，室温反应1小时。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-((tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid Trifluoroacetic acid (1.0 ml) was added to a di-tert-butyl ester (153 mg, 0.16 mmol) solution in dichloromethane (4.0 ml), after the addition, the reaction was carried out at room temperature for 1 hour.
减压浓缩，所得粗产物溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到98毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(四氢呋喃-3-基)甲基-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸三氟乙酸盐。(收率67.2％)。LCMS:RT＝2.81min,[M-H] -＝779.13。 1H NMR(400MHz,DMSO)δ11.74(s,1H),10.15(s,1H),9.77(s,1H),8.00(s,1H),7.94(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.2Hz,1H),7.43–7.26(m,6H),7.07(d,J＝1.5Hz,1H),5.46–5.30(m,1H),4.23–3.52(m,15H),3.45(t,J＝8.4Hz,1H),3.36(dd,J＝14.4,5.2Hz,1H),3.14(dd,J＝14.8,10.0Hz,1H),2.74-2.67(m,1H),2.15-2.06(m,1H),1.69-1.59(m,1H)。 Concentrated under reduced pressure, the obtained crude product was dissolved in dichloromethane (1.0 ml), and added dropwise to n-hexane (10.0 ml). A white solid precipitated out. The filter cake was washed with n-hexane and dried to obtain 98 mg of white solid. (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-(4-(Tetrahydrofuran-3-yl)methyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid trifluoroacetate. (Yield 67.2%). LCMS: RT = 2.81 min, [MH] - =779.13. 1 H NMR (400MHz, DMSO) δ 11.74 (s, 1H), 10.15 (s, 1H), 9.77 (s, 1H), 8.00 (s, 1H), 7.94 (d, J = 2.2 Hz, 1H), 7.82(d,J=8.6Hz,1H),7.77(dd,J=8.6,2.2Hz,1H),7.43-7.26(m,6H),7.07(d,J=1.5Hz,1H),5.46-5.30 (m, 1H), 4.23–3.52 (m, 15H), 3.45 (t, J = 8.4 Hz, 1H), 3.36 (dd, J = 14.4, 5.2 Hz, 1H), 3.14 (dd, J = 14.8, 10.0 Hz, 1H), 2.74-2.67 (m, 1H), 2.15-2.06 (m, 1H), 1.69-1.59 (m, 1H).
实施例22Example 22
合成(S)-5-(3-(4-(4-(羧基甲氧基)哌啶-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(3-(4-(4-(carboxymethoxy)piperidine-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H- Tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000113
Figure PCTCN2020130361-appb-000113
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(3-(4-(4-(2-(叔丁氧基)-2-氧代乙氧基)哌啶-1-甲酰胺基)苯基)-2-(4-(5-)氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(3-(4-(4-(2-(tert-butoxy)-2-oxoethoxy)piperidine-1-carboxamido)phenyl)- 2-(4-(5-)chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole -1,2-Di-tert-butyl dicarboxylic acid
Figure PCTCN2020130361-appb-000114
Figure PCTCN2020130361-appb-000114
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(100毫克，0.11毫摩尔)和叔丁基哌啶-4-基碳酸酯(277毫克，1.4毫摩尔)的四氢呋喃(6.0毫升)中滴加N,N-二异丙基乙胺(284毫克，2.2毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (100 mg, 0.11 mmol) And tert-butylpiperidin-4-yl carbonate (277 mg, 1.4 mmol) in tetrahydrofuran (6.0 ml) was added dropwise N,N-diisopropylethylamine (284 mg, 2.2 mmol), the drop was completed , Reaction at 60 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：正己烷/乙酸乙酯＝1/3)。得到50毫克白色固体(S)-5-(3-(4-(4-(2-(叔丁氧基)-2-氧代乙氧基)哌啶-1-甲酰胺基)苯基)-2-(4-(5-)氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：45.0％)。LCMS:RT＝4.48min,[M+H] +＝1011.4。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: n-hexane/ethyl acetate = 1/3). Obtain 50 mg of white solid (S)-5-(3-(4-(4-(2-(tert-butoxy)-2-oxoethoxy)piperidine-1-carboxamido)phenyl) -2-(4-(5-)chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole Dole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 45.0%). LCMS: RT = 4.48 min, [M+H] + =1011.4.
步骤B：合成(S)-5-(3-(4-(4-(羧基甲氧基)哌啶-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(3-(4-(4-(carboxymethoxy)piperidine-1-carboxamido)phenyl)-2-(4-(5-chloro-2- (1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000115
Figure PCTCN2020130361-appb-000115
室温下，将(S)-5-(3-(4-(4-(2-(叔丁氧基)-2-氧代乙氧基)哌啶-1-甲酰胺基)苯基)-2-(4-(5-)氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(50毫克，0.05毫摩尔)加入二氯甲烷(5.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。At room temperature, (S)-5-(3-(4-(4-(2-(tert-butoxy)-2-oxoethoxy)piperidine-1-carboxamido)phenyl)- 2-(4-(5-)chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole Di-tert-butyl 1,2-dicarboxylate (50 mg, 0.05 mmol) was added to dichloromethane (5.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到35毫克白色固体(S)-5-(3-(4-(4-(羧基甲氧基)哌啶-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸(收率：91.0％)。LCMS:RT＝3.25min,[M-H] -＝797.26。 1H NMR(400MHz,DMSO)δ12.74(s,2H),11.71(s,1H),10.13(s,1H),9.78(s, 1H),8.47(s,1H),8.00(s,1H),7.95(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.2Hz,1H),7.50–7.35(m,3H),7.32(dd,J＝8.9,1.8Hz,1H),7.14(d,J＝8.1Hz,2H),7.06(d,J＝1.6Hz,1H),5.30(dd,J＝9.8,5.8Hz,1H),4.06(s,2H),3.97–3.67(m,5H),3.61–3.52(m,2H),3.30–3.18(m,1H),3.17–2.91(m,3H),1.85(dd,J＝11.2,6.7Hz,2H),1.42(dd,J＝14.0,6.9Hz,2H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 35 mg of white solid (S)-5-(3-(4-(4-(carboxymethoxy)piperidine-1-carboxamido)phenyl)-2-( 4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxy Acid (yield: 91.0%). LCMS: RT = 3.25 min, [MH] - = 797.26. 1 H NMR (400MHz, DMSO) δ 12.74 (s, 2H), 11.71 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.47 (s, 1H), 8.00 (s, 1H) ), 7.95 (d, J = 2.2 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.76 (dd, J = 8.6, 2.2 Hz, 1H), 7.50–7.35 (m, 3H), 7.32 (dd,J=8.9,1.8Hz,1H), 7.14(d,J=8.1Hz,2H), 7.06(d,J=1.6Hz,1H), 5.30(dd,J=9.8,5.8Hz,1H) ,4.06(s,2H),3.97–3.67(m,5H),3.61–3.52(m,2H), 3.30–3.18(m,1H),3.17–2.91(m,3H),1.85(dd,J= 11.2, 6.7 Hz, 2H), 1.42 (dd, J=14.0, 6.9 Hz, 2H).
实施例23Example 23
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟乙基)氨基甲酰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000116
Figure PCTCN2020130361-appb-000116
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟乙基)氨甲酰基)哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxy Di-tert-butyl ester
Figure PCTCN2020130361-appb-000117
Figure PCTCN2020130361-appb-000117
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(133毫克，0.15毫摩尔)和N-(2-羟乙基)哌啶-4-甲酰胺盐酸盐(312毫克，1.5毫摩尔)的四氢呋喃(6.0毫升)中滴加N,N-二异丙基乙胺(387毫克，3.0毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (133 mg, 0.15 mmol) And N-(2-hydroxyethyl)piperidine-4-carboxamide hydrochloride (312 mg, 1.5 mmol) in tetrahydrofuran (6.0 ml) was added dropwise N,N-diisopropylethylamine (387 mg , 3.0 mmol), after dripping, react at 60 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：DCM/MeOH＝10/1)。得到55.3毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟乙基)氨甲酰基)哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：38.1％)。LCMS:RT＝4.02min,[M+H] +＝968.23。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: DCM/MeOH=10/1). Obtain 55.3 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-di Di-tert-butyl carboxylate (yield: 38.1%). LCMS: RT = 4.02 min, [M+H] + =968.23.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟乙基)氨基甲酰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000118
Figure PCTCN2020130361-appb-000118
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟乙基)氨甲酰基)哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(53.3毫克，0.06毫摩尔)加入二氯甲烷(4.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxy Di-tert-butyl acid (53.3 mg, 0.06 mmol) was added to dichloromethane (4.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷 (10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到42.7毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟乙基)氨基甲酰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：95.7％。LCMS:RT＝3.07min,[M+H] +＝812.30。 1H NMR(500MHz,DMSO)δ10.59(s,1H),9.40(s,1H),7.81(s,1H),7.47(s,1H),6.13(s,1H),5.69(s,1H),5.64(d,J＝2.2Hz,1H),5.51(d,J＝8.6Hz,1H),5.49–5.43(m,1H),5.09(t,J＝7.5Hz,3H),5.06(s,2H),5.02(dd,J＝9.0,1.8Hz,2H),4.84(d,J＝7.8Hz,1H),4.75(d,J＝1.7Hz,1H),3.44(s,1H),2.99(dd,J＝9.7,5.9Hz,1H),2.31(t,J＝5.3Hz,3H),1.80(d,J＝13.8Hz,3H),1.43(s,2H),1.07(dd,J＝11.8,6.0Hz,1H),0.97–0.88(m,1H),0.82–0.77(m,2H),0.70(dd,J＝14.2,9.3Hz,2H),0.47(t,J＝12.2Hz,3H),0.23(s,1H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 42.7 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H- Indole-2-carboxylic acid (yield: 95.7%. LCMS: RT = 3.07 min, [M+H] + = 812.30. 1 H NMR (500MHz, DMSO) δ 10.59 (s, 1H), 9.40 (s ,1H),7.81(s,1H),7.47(s,1H),6.13(s,1H),5.69(s,1H), 5.64(d,J=2.2Hz,1H),5.51(d,J= 8.6Hz,1H),5.49–5.43(m,1H),5.09(t,J=7.5Hz,3H),5.06(s,2H),5.02(dd,J=9.0,1.8Hz,2H),4.84( d,J=7.8Hz,1H), 4.75(d,J=1.7Hz,1H), 3.44(s,1H), 2.99(dd,J=9.7,5.9Hz,1H), 2.31(t,J=5.3 Hz, 3H), 1.80 (d, J = 13.8 Hz, 3H), 1.43 (s, 2H), 1.07 (dd, J = 11.8, 6.0 Hz, 1H), 0.97-0.88 (m, 1H), 0.82-0.77 (m, 2H), 0.70 (dd, J=14.2, 9.3 Hz, 2H), 0.47 (t, J=12.2 Hz, 3H), 0.23 (s, 1H).
实施例24Example 24
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟乙基)(甲基)氨基甲酰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000119
Figure PCTCN2020130361-appb-000119
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟基乙基)(甲基)氨基甲酰基)哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1, Di-tert-butyl 2-dicarboxylate
Figure PCTCN2020130361-appb-000120
Figure PCTCN2020130361-appb-000120
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(89毫克，0.1毫摩尔)和N-(2-羟乙基)-N-甲基哌啶-4-甲酰胺(300毫克，1.0毫摩尔)的四氢呋喃(6.0毫升)中滴加N,N-二异丙基乙胺(258毫克，2.0毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (89 mg, 0.1 mmol) And N-(2-hydroxyethyl)-N-methylpiperidine-4-carboxamide (300 mg, 1.0 mmol) in tetrahydrofuran (6.0 ml) was added dropwise N,N-diisopropylethylamine ( 258 mg, 2.0 mmol), after dripping, react overnight at 60 degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：DCM/MeOH＝10/1)。得到42.2毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟基乙基)(甲基)氨基甲酰基)哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：43.2％)。LCMS:RT＝3.39min,[M+H] +＝982.38。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: DCM/MeOH=10/1). Obtain 42.2 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1 , Di-tert-butyl 2-dicarboxylic acid (yield: 43.2%). LCMS: RT = 3.39 min, [M+H] + =982.38.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟乙基)(甲基)氨基甲酰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2- carboxylic acid
Figure PCTCN2020130361-appb-000121
Figure PCTCN2020130361-appb-000121
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟基乙基)(甲基)氨基甲酰基) 哌啶-1-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(42.2毫克，0.04毫摩尔)加入二氯甲烷(4.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1, Di-tert-butyl 2-dicarboxylate (42.2 mg, 0.04 mmol) was added to dichloromethane (4.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到31.8毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-((2-羟乙基)(甲基)氨基甲酰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：96.1％)。LCMS:RT＝2.75min,[M-H] -＝826.23。 1H NMR(500MHz,DMSO)δ12.90(s,1H),11.72(s,1H),10.14(s,1H),9.80(s,1H),8.56–8.21(m,3H),8.02(s,1H),7.96(t,J＝2.3Hz,1H),7.84(dd,J＝8.6,4.5Hz,1H),7.78(dt,J＝8.6,2.6Hz,1H),7.47–7.37(m,3H),7.35(dd,J＝8.6,2.7Hz,1H),7.18(t,J＝14.1Hz,2H),7.09(t,J＝8.4Hz,1H),5.41–5.19(m,2H),4.32–4.18(m,1H),4.05(dd,J＝24.7,16.2Hz,3H),3.75(s,2H),3.66–3.52(m,2H),2.67–2.56(m,3H),2.05–1.93(m,2H),1.87(dd,J＝30.9,12.8Hz,2H),1.74–1.61(m,1H),1.62–1.41(m,3H),0.87(t,J＝6.9Hz,1H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 31.8 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenyl)propionamido ) -1H-Indole-2-carboxylic acid (yield: 96.1%). LCMS: RT = 2.75 min, [MH] - = 826.23. 1 H NMR (500MHz, DMSO) δ 12.90 (s, 1H), 11.72 (s, 1H), 10.14 (s, 1H), 9.80 (s, 1H), 8.56-8.21 (m, 3H), 8.02 (s ,1H),7.96(t,J=2.3Hz,1H),7.84(dd,J=8.6,4.5Hz,1H),7.78(dt,J=8.6,2.6Hz,1H),7.47–7.37(m, 3H), 7.35 (dd, J = 8.6, 2.7 Hz, 1H), 7.18 (t, J = 14.1 Hz, 2H), 7.09 (t, J = 8.4 Hz, 1H), 5.41-5.19 (m, 2H), 4.32–4.18(m,1H),4.05(dd,J=24.7,16.2Hz,3H), 3.75(s,2H), 3.66–3.52(m,2H), 2.67–2.56(m,3H), 2.05– 1.93 (m, 2H), 1.87 (dd, J = 30.9, 12.8 Hz, 2H), 1.74-1.61 (m, 1H), 1.62-1.41 (m, 3H), 0.87 (t, J = 6.9 Hz, 1H) .
实施例25Example 25
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000122
Figure PCTCN2020130361-appb-000122
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(4-Hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl
Figure PCTCN2020130361-appb-000123
Figure PCTCN2020130361-appb-000123
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(133毫克，0.15毫摩尔)和3-氨基-3-甲基丁-1-醇(154毫克，0.75毫摩尔)的四氢呋喃(6.0毫升)中滴加N,N-二异丙基乙胺(387毫克，1.5毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (133 mg, 0.15 mmol) And 3-amino-3-methylbutan-1-ol (154 mg, 0.75 mmol) in tetrahydrofuran (6.0 mL) was added dropwise N,N-diisopropylethylamine (387 mg, 1.5 mmol), After dripping, react overnight at 60 degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：DCM/MeOH＝10/1)。得到56.6毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：38.4％)。LCMS:RT＝3.39min,[M+H] +＝982.38。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: DCM/MeOH=10/1). Obtain 56.6 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (yield: 38.4%). LCMS: RT = 3.39 min, [M+H] + =982.38.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000124
Figure PCTCN2020130361-appb-000124
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(55.6毫克，0.06毫摩尔)加入二氯甲烷(4.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(4-Hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl ester (55.6 mg, 0.06 mmol) was added to methylene chloride (4.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到43.1毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：92.4％)。LCMS:RT＝3.28min,[M-H] -＝741.15。 1H NMR(500MHz,DMSO)δ12.92(s,1H),11.72(s,1H),10.13(s,1H),9.80(s,1H),8.27(s,1H),8.04–7.94(m,2H),7.83(d,J＝8.6Hz,1H),7.78(dd,J＝8.6,2.3Hz,1H),7.39(d,J＝8.8Hz,1H),7.37–7.29(m,2H),7.10(dd,J＝27.1,4.3Hz,3H),5.98(s,1H),5.29(dd,J＝9.9,5.7Hz,2H),4.38(t,J＝4.9Hz,1H),3.74(s,2H),3.51(dd,J＝12.0,7.0Hz,2H),3.27–3.15(m,1H),3.01(dd,J＝15.8,10.4Hz,3H),1.82(t,J＝7.1Hz,2H),1.27(d,J＝13.5Hz,6H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 43.1 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole- 2-carboxylic acid (yield: 92.4%). LCMS: RT = 3.28 min, [MH]- =741.15. 1 H NMR (500MHz, DMSO) δ 12.92 (s, 1H), 11.72 (s, 1H), 10.13 (s, 1H), 9.80 (s, 1H), 8.27 (s, 1H), 8.04-7.94 (m ,2H),7.83(d,J=8.6Hz,1H),7.78(dd,J=8.6,2.3Hz,1H),7.39(d,J=8.8Hz,1H),7.37–7.29(m,2H) ,7.10(dd,J=27.1,4.3Hz,3H), 5.98(s,1H), 5.29(dd,J=9.9,5.7Hz,2H), 4.38(t,J=4.9Hz,1H), 3.74( s, 2H), 3.51 (dd, J = 12.0, 7.0 Hz, 2H), 3.27–3.15 (m, 1H), 3.01 (dd, J = 15.8, 10.4 Hz, 3H), 1.82 (t, J = 7.1 Hz , 2H), 1.27 (d, J=13.5 Hz, 6H).
实施例26Example 26
合成5–((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2R，3R)-1,3-二羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((2R,3R)-1,3-dihydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000125
Figure PCTCN2020130361-appb-000125
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5–((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S，3S)-1,3-二羟基丁-2-基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2S,3S)-1,3-dihydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
Figure PCTCN2020130361-appb-000126
Figure PCTCN2020130361-appb-000126
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(78毫克，0.2毫摩尔)和(2R,3S)-2-氨基丁烷-1,3-二醇(210毫克，2.0毫摩尔)的四氢呋喃(6.0毫升)中滴加N,N-二异丙基乙胺(5.6毫克，0.04毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (78 mg, 0.2 mmol) And (2R,3S)-2-aminobutane-1,3-diol (210mg, 2.0mmol) in tetrahydrofuran (6.0ml) was added dropwise N,N-diisopropylethylamine (5.6mg, 0.04 mmol), after dripping, react overnight at 60 degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：DCM/MeOH＝10/1)。得到65.7毫克白色固体5–((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S，3S)-1,3-二羟基丁-2-基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：36.4％)。LCMS:RT＝4.05min,[M+H] +＝901.33。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: DCM/MeOH=10/1). 65.7 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((2S,3S)-1,3-dihydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2- Di-tert-butyl dicarboxylate (yield: 36.4%). LCMS: RT = 4.05 min, [M+H] + =901.33.
步骤B：合成5–((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2R，3R)-1,3-二羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2R,3R)-1,3-dihydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000127
Figure PCTCN2020130361-appb-000127
室温下，将5–((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S，3S)-1,3-二羟基丁-2-基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(65.7毫克，0.07毫摩尔)加入二氯甲烷(4.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。At room temperature, add 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2S,3S)-1,3-dihydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate (65.7 mg, 0.07 mmol) was added to dichloromethane (4.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到52.4毫克白色固体5–((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2R，3R)-1,3-二羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：96.5％)。LCMS:RT＝3.09min,[M+H] +＝745.27。 1H NMR(400MHz,DMSO)δ12.93(s,2H),11.73(s,1H),10.15(s,1H),9.80(s,1H),8.69(s,1H),8.01(s,1H),7.96(d,J＝2.3Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.41–7.30(m,3H),7.26–7.02(m,3H),5.97(d,J＝8.3Hz,1H),5.36–5.19(m,1H),3.97(dt,J＝14.1,7.1Hz,2H),3.39(d,J＝11.8Hz,3H),3.25–3.12(m,1H),3.09–2.92(m,1H),1.99(dt,J＝7.9,2.2Hz,1H),1.24(s,4H),1.06(d,J＝6.4Hz,2H),0.86(t,J＝5.4Hz,1H)。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 52.4 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-((2R,3R)-1,3-dihydroxybut-2-yl)ureido)phenyl)propionamido)-1H -Indole-2-carboxylic acid (yield: 96.5%). LCMS: RT = 3.09 min, [M+H] + =745.27. 1 H NMR (400MHz, DMSO) δ 12.93 (s, 2H), 11.73 (s, 1H), 10.15 (s, 1H), 9.80 (s, 1H), 8.69 (s, 1H), 8.01 (s, 1H) ),7.96(d,J=2.3Hz,1H),7.83(d,J=8.6Hz,1H),7.77(dd,J=8.6,2.3Hz,1H),7.41-7.30(m,3H),7.26 –7.02(m,3H),5.97(d,J=8.3Hz,1H),5.36–5.19(m,1H),3.97(dt,J＝14.1,7.1Hz,2H), 3.39(d,J＝11.8 Hz, 3H), 3.25–3.12 (m, 1H), 3.09–2.92 (m, 1H), 1.99 (dt, J = 7.9, 2.2 Hz, 1H), 1.24 (s, 4H), 1.06 (d, J = 6.4Hz, 2H), 0.86 (t, J=5.4Hz, 1H).
实施例27Example 27
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000128
Figure PCTCN2020130361-appb-000128
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-2-氨基-3-(4-溴苯基)丙酸叔丁酯Step A: Synthesis of tert-butyl (S)-2-amino-3-(4-bromophenyl)propionate
Figure PCTCN2020130361-appb-000129
Figure PCTCN2020130361-appb-000129
将(S)-2-氨基-3-(4-溴苯基)丙酸(10.0克，41.1毫摩尔)溶于丙酸叔丁酯(70.0毫升)的溶液中。随后，向上述溶液中加入70％质量分数的高氯酸(6.1克，61.7毫摩尔)，置换氮气3遍。在室温下搅拌过夜。(S)-2-amino-3-(4-bromophenyl)propionic acid (10.0 g, 41.1 mmol) was dissolved in a solution of tert-butyl propionate (70.0 mL). Subsequently, 70% mass fraction of perchloric acid (6.1 g, 61.7 mmol) was added to the above solution, and nitrogen was replaced 3 times. Stir overnight at room temperature.
将反应液缓慢滴加到饱和碳酸氢钠溶液(250毫升)淬灭反应。混合液用乙酸乙酯(200毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(100毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到11.5克黄色油状(S)-2-氨基-3-(4-溴苯基)丙酸叔丁酯(收率：93.50％)。LCMS：RT＝2.80min，[M+H] +＝300.05。 The reaction solution was slowly added dropwise to saturated sodium bicarbonate solution (250 mL) to quench the reaction. The mixture was extracted with ethyl acetate (200 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (100 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. 11.5 g of tert-butyl (S)-2-amino-3-(4-bromophenyl)propionate was obtained as a yellow oil (yield: 93.50%). LCMS: RT = 2.80 min, [M+H] + =300.05.
步骤B：合成(S)-3-(4-溴苯基)-2-((叔丁氧基羰基)氨基)丙酸叔丁酯Step B: Synthesis of (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid tert-butyl ester
Figure PCTCN2020130361-appb-000130
Figure PCTCN2020130361-appb-000130
将(S)-2-氨基-3-(4-溴苯基)丙酸叔丁酯(11.50克，38.3毫摩尔)和氢氧化钠(3.16克，76.6毫摩尔)溶于四氢呋喃(20毫升)和水(10毫升)的混合溶液中。随后，向上述溶液中加入二碳酸二叔丁酯(10.30克,4.1毫摩尔)。在室温下搅拌1小时。(S)-2-amino-3-(4-bromophenyl)-tert-butyl propionate (11.50 g, 38.3 mmol) and sodium hydroxide (3.16 g, 76.6 mmol) were dissolved in tetrahydrofuran (20 mL) And water (10 ml) mixed solution. Subsequently, di-tert-butyl dicarbonate (10.30 g, 4.1 mmol) was added to the above solution. Stir at room temperature for 1 hour.
向反应液中加入饱和氯化铵溶液(200毫升)。混合液用乙酸乙酯(100毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(50毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/10)。得到9.80克固体(S)-3-(4-溴苯基)-2-((叔丁氧基羰基)氨基)丙酸叔丁酯(收率：64.0％)。Saturated ammonium chloride solution (200 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (100 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (50 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/10). 9.80 g of solid tert-butyl (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionate was obtained (yield: 64.0%).
步骤C：合成4-异丙基哌嗪-2-酮Step C: Synthesis of 4-isopropylpiperazin-2-one
Figure PCTCN2020130361-appb-000131
Figure PCTCN2020130361-appb-000131
将哌嗪-2-酮(11.0克，55.5毫摩尔)溶于甲醇(20.0毫升)和乙酸(0.5毫升)的混合溶液中。随后，向上述溶液中加入丙酮(6.78克，111.0毫摩尔)和氰基硼氢化钠(4.19克，66.6毫摩尔)。在60摄氏度下搅拌过夜。Piperazine-2-one (11.0 g, 55.5 mmol) was dissolved in a mixed solution of methanol (20.0 mL) and acetic acid (0.5 mL). Subsequently, acetone (6.78 g, 111.0 mmol) and sodium cyanoborohydride (4.19 g, 66.6 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
将反应液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到3.00克白色固体4-异丙基哌嗪-2-酮(收率：38.0％)。LCMS：RT＝0.83min，[M+H] +＝143.11。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 3.00 g of white solid 4-isopropylpiperazin-2-one was obtained (yield: 38.0%). LCMS: RT=0.83 min, [M+H] + =143.11.
步骤D：合成(S)-2-((叔丁氧基羰基)氨基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionic acid tert Butyl
Figure PCTCN2020130361-appb-000132
Figure PCTCN2020130361-appb-000132
将(S)-3-(4-溴苯基)-2-((叔丁氧基羰基)氨基)丙酸叔丁酯(8.00克，20.0毫摩尔)和4-异丙基哌嗪-2-酮(5.68克，40.0摩尔)溶于甲苯(100毫升)的溶液中。随后，向上述溶液中加入N，N-二甲基乙-1,2-二胺(3.50克,40.0毫摩尔)，碘化亚铜(3.80克，20.0毫摩尔)和碳酸铯(13.00克，40.0毫摩尔)。在110摄氏度下搅拌18个小时。Combine (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid tert-butyl ester (8.00 g, 20.0 mmol) and 4-isopropylpiperazine-2 -Ketone (5.68 g, 40.0 mol) is dissolved in a solution of toluene (100 ml). Subsequently, N,N-dimethylethyl-1,2-diamine (3.50 g, 40.0 mmol), cuprous iodide (3.80 g, 20.0 mmol) and cesium carbonate (13.00 g, 40.0 mmol). Stir at 110 degrees Celsius for 18 hours.
向反应液中加入饱和氯化铵溶液(200毫升)。混合液用乙酸乙酯(150毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(100毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/10)。得到3.25克黄色固体(S)-2-((叔丁氧基羰基)氨基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：35.0％)。LCMS：RT＝3.55min，[M+H] +＝462.30。 Saturated ammonium chloride solution (200 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (150 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (100 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/10). Obtain 3.25 g of yellow solid (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionic acid Tert-butyl ester (yield: 35.0%). LCMS: RT = 3.55 min, [M+H] + =462.30.
步骤E：合成(S)-2-氨基-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯盐酸盐Step E: Synthesis of (S)-2-amino-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionic acid tert-butyl ester hydrochloride
Figure PCTCN2020130361-appb-000133
Figure PCTCN2020130361-appb-000133
将(S)-2-((叔丁氧基羰基)氨基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(4.00克，8.67毫摩尔)溶于乙酸乙酯(35.0毫升)中。随后，向上述溶液中加入盐酸乙酸乙酯溶液(2摩尔/升，8.6毫升,17.36毫摩尔)。在室温下搅拌8小时。Add (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate ( 4.00 g, 8.67 mmol) was dissolved in ethyl acetate (35.0 ml). Subsequently, a hydrochloric acid ethyl acetate solution (2 mol/L, 8.6 mL, 17.36 mmol) was added to the above solution. Stir at room temperature for 8 hours.
将反应液减压浓缩。得到2.50克固体(S)-2-氨基-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯盐酸盐(收率：80.0％)。The reaction solution was concentrated under reduced pressure. Obtained 2.50 g of solid tert-butyl (S)-2-amino-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionate hydrochloride (yield: 80.0%).
步骤F：合成(S)-2-(2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step F: Synthesis of (S)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido)-3-(4-(4- Isopropyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000134
Figure PCTCN2020130361-appb-000134
将(S)-2-氨基-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯盐酸盐(2.00克，5.5毫摩尔)和2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酸(1.96克，6.6摩尔)溶于N,N-二甲基甲酰胺(20.0毫升)的溶液中。随后，向上述溶液中加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(3.20克,8.3毫摩尔)，N,N-二异丙基乙胺(1.40克，11.0毫摩尔)，在室温下搅拌18个小时。(S)-2-amino-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate hydrochloride (2.00 g, 5.5 mmol ) And 2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetic acid (1.96g, 6.6mol) dissolved in N,N-dimethylformamide (20.0 Ml) of the solution. Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (3.20 g, 8.3 mmol) was added to the above solution, N, N -Diisopropylethylamine (1.40 g, 11.0 mmol), stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(100毫升)。混合液用乙酸乙酯(50毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(100毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到1.80克白色固体(S)-2-(2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：50.0％)。LCMS：RT＝3.65min，[M+H] +＝638.30。 Saturated ammonium chloride solution (100 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (50 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (100 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 1.80 g of white solid (S)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido)-3-(4-(4 -Isopropyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate (yield: 50.0%). LCMS: RT = 3.65 min, [M+H] + =638.30.
步骤G：合成(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪)-叔丁基嗪-1-基)苯基)丙酸叔丁酯Step G: Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-isopropyl-2-oxopiperazine)-tert-butylazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000135
Figure PCTCN2020130361-appb-000135
将(S)-2-(2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)叔丁基基)苯基)丙酸叔丁酯(1.80克，2.8毫摩尔)和1,2-二溴乙烷(3.70克，19.7毫摩尔)溶于乙腈(20.0毫升)中。随后，向上述溶液中加入碳酸钾(2.70克，19.7毫摩尔)。在90摄氏度下搅拌过夜。(S)-2-(2-((5-Chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido)-3-(4-(4-isopropyl -2-oxopiperazin-1-yl)tert-butyl)phenyl)propionate (1.80 g, 2.8 mmol) and 1,2-dibromoethane (3.70 g, 19.7 mmol) Dissolve in acetonitrile (20.0 mL). Subsequently, potassium carbonate (2.70 g, 19.7 mmol) was added to the above solution. Stir overnight at 90 degrees Celsius.
将反应液过滤，滤饼用乙腈(50毫升×3次)洗涤。滤液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到500毫克白色油状物(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪)-叔丁基嗪-1-基)苯基)丙酸叔丁酯(收率：37.0％)。LCMS：RT＝3.30min，[M+H] +＝664.32。 The reaction solution was filtered, and the filter cake was washed with acetonitrile (50 ml×3 times). The filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 500 mg of white oil (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-(4-isopropyl-2-oxopiperazine)-tert-butylazin-1-yl)phenyl) tert-butyl propionate (yield: 37.0%). LCMS: RT = 3.30 min, [M+H] + =664.32.
步骤H：合成(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step H: Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-isopropyl 2-oxopiperazin-1-yl) phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000136
Figure PCTCN2020130361-appb-000136
将(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪)-1-基)苯基)丙酸叔丁酯(500毫克，0.86毫摩尔)和1,3-二甲基嘧啶-2,4,6(1H,3H,5H)-三酮烷(587毫克，4.30毫摩尔)溶于二氯甲烷(10.0毫升)中。随后，向上述溶液中加入四(三苯基膦)钯(321毫克，0.05毫摩尔)。在40摄氏度下搅拌过夜。Add (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4 -Isopropyl-2-oxopiperazine)-1-yl)phenyl)propionic acid tert-butyl ester (500 mg, 0.86 mmol) and 1,3-dimethylpyrimidine-2,4,6 (1H ,3H,5H)-Trione alkane (587 mg, 4.30 mmol) was dissolved in dichloromethane (10.0 mL). Subsequently, tetrakis(triphenylphosphine)palladium (321 mg, 0.05 mmol) was added to the above solution. Stir overnight at 40 degrees Celsius.
向反应液中加入饱和碳酸氢钠溶液(20毫升)。混合液用乙酸乙酯(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(20毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂： 甲醇/二氯甲烷＝1/10)。得到290毫克黄色固体(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：64.0％)。LCMS：RT＝2.91min，[M+H] +＝566.22。 Saturated sodium bicarbonate solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (20 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 290 mg of yellow solid (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-iso Propyl-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate (yield: 64.0%). LCMS: RT = 2.91 min, [M+H] + =566.22.
步骤I：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step I: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000137
Figure PCTCN2020130361-appb-000137
将(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(150毫克，0.25毫摩尔)和原甲酸三乙酯(190毫克，1.28毫摩尔)溶于醋酸(5.0毫升)中。随后，向上述溶液中加入叠氮化钠(83毫克，1.28毫摩尔)。在70摄氏度下搅拌1小时。(S)-2-(4-(2-Amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-isopropyl-2 -Oxopiperazin-1-yl)phenyl) tert-butyl propionate (150 mg, 0.25 mmol) and triethyl orthoformate (190 mg, 1.28 mmol) were dissolved in acetic acid (5.0 mL). Subsequently, sodium azide (83 mg, 1.28 mmol) was added to the above solution. Stir at 70 degrees Celsius for 1 hour.
向反应液中加入饱和碳酸氢钠溶液(20毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到120毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：73.0％)。LCMS：RT＝2.90min，[M+H] +＝637.24。 Saturated sodium bicarbonate solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 120 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-Butyl 3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionate (yield: 73.0%). LCMS: RT = 2.90 min, [M+H] + =637.24.
步骤J：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸Step J: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionic acid
Figure PCTCN2020130361-appb-000138
Figure PCTCN2020130361-appb-000138
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(120毫克，0.20毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-Isopropyl-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (120 mg, 0.20 mmol) was dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。得到100毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸(收率：91.0％)。The reaction solution was concentrated under reduced pressure. Obtain 100 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- 3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionic acid (yield: 91.0%).
步骤K：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step K: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000139
Figure PCTCN2020130361-appb-000139
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸(100毫克，0.2毫摩尔)和5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯(69毫克，0.2摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)的溶液 中。随后，向上述溶液中加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(98毫克，0.3毫摩尔)，N,N-二异丙基乙胺(44毫克，0.4毫摩尔)，在室温下搅拌18个小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionic acid (100 mg, 0.2 mmol) and 5-amino-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (69 mg, 0.2 mol) was dissolved in a solution of N,N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (98 mg, 0.3 mmol) was added to the above solution, N, N -Diisopropylethylamine (44 mg, 0.4 mmol), stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(10毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到115毫克白色油状物(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：75.0％)。LCMS：RT＝3.44min，[M+H] +＝895.97。 Saturated ammonium chloride solution (10 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 115 mg of white oil (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1 -Yl)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl Ester (yield: 75.0%). LCMS: RT = 3.44 min, [M+H] + =895.97.
步骤L：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step L: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000140
Figure PCTCN2020130361-appb-000140
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-二叔丁酯(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(115毫克，0.10毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Di-tert-butyl ester (4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (115 mg, 0.10 mmol) was dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。所得残余物用制备HPLC纯化得到57.6毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：60.0％)。LCMS：RT＝2.71min，[M+H] +＝739.40。 The reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 57.6 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-di Oxopiperazin-1-yl)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid (Yield: 60.0%). LCMS: RT = 2.71 min, [M+H] + =739.40.
实施例28Example 28
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000141
Figure PCTCN2020130361-appb-000141
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-3-(4-溴苯基)-2-(2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酰氨基)丙酸叔丁酯Step A: Synthesis of (S)-3-(4-bromophenyl)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido ) Tert-butyl propionate
Figure PCTCN2020130361-appb-000142
Figure PCTCN2020130361-appb-000142
将(S)-2-氨基-3-(4-溴苯基)丙酸叔丁酯(10.0克，33毫摩尔)和2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酸(10.8克，36毫摩尔)溶于N,N-二甲基甲酰胺(100.0毫升)的溶液中。随后，向上述溶液中加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(19.0克，49毫摩尔)，N,N-二异丙基乙胺(8.6毫克，66毫摩尔)，在室温下搅拌18个小时。Combine (S)-2-amino-3-(4-bromophenyl) tert-butyl propionate (10.0 g, 33 mmol) and 2-((5-chloro-2-(diallylamino)benzene (Yl)amino)-2-oxoacetic acid (10.8 g, 36 mmol) was dissolved in a solution of N,N-dimethylformamide (100.0 ml). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (19.0 g, 49 mmol) was added to the above solution, N, N -Diisopropylethylamine (8.6 mg, 66 mmol), stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(200毫升)。混合液用乙酸乙酯(100毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(100毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到12克白色固体(S)-3-(4-溴苯基)-2-(2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酰氨基)丙酸叔丁酯(收率：62.0％)。LCMS：RT＝3.90min，[M+H] +＝576.12。 Saturated ammonium chloride solution (200 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (100 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (100 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 12 grams of white solid (S)-3-(4-bromophenyl)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetyl (Amino) tert-butyl propionate (yield: 62.0%). LCMS: RT = 3.90 min, [M+H] + =576.12.
步骤B：合成(S)-3-(4-溴苯基)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯Step B: Synthesis of (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazine -1-yl) tert-butyl propionate
Figure PCTCN2020130361-appb-000143
Figure PCTCN2020130361-appb-000143
将(S)-3-(4-溴苯基)-2-(2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酰氨基)丙酸叔丁酯(12.0克，20.8毫摩尔)溶于乙腈(100.0毫升)的溶液中。随后，向上述溶液中加入1,2-二溴乙烷(27.0克，148.0毫摩尔)，碳酸钾(20.0克，145.0毫摩尔)，在90摄氏度下搅拌18个小时。(S)-3-(4-Bromophenyl)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido)propionic acid Tert-butyl ester (12.0 g, 20.8 mmol) was dissolved in a solution of acetonitrile (100.0 ml). Subsequently, 1,2-dibromoethane (27.0 g, 148.0 mmol) and potassium carbonate (20.0 g, 145.0 mmol) were added to the above solution, and stirred at 90 degrees Celsius for 18 hours.
将反应液冷却到室温，过滤。滤液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到3.7克无色油状(S)-3-(4-溴苯基)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(收率：29.5％)。LCMS：RT＝3.85min，[M+H] +＝602.13。 The reaction solution was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 3.7 g of colorless oil (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxo Piperazin-1-yl) tert-butyl propionate (yield: 29.5%). LCMS: RT = 3.85 min, [M+H] + =602.13.
步骤C：合成4-(4-甲基环己基)哌嗪-2-酮Step C: Synthesis of 4-(4-methylcyclohexyl)piperazin-2-one
Figure PCTCN2020130361-appb-000144
Figure PCTCN2020130361-appb-000144
将哌嗪-2-酮(1.0克，9.9毫摩尔)溶于甲醇(10.0毫升)和乙酸(0.2毫升)的混合溶液中。随后，向上述溶液中加入4-甲基环己-1-酮(2.6克，19.9毫摩尔)和氰基硼氢化钠(1.3克，19.9毫摩尔)。在60摄氏度下搅拌过夜。Piperazine-2-one (1.0 g, 9.9 mmol) was dissolved in a mixed solution of methanol (10.0 mL) and acetic acid (0.2 mL). Subsequently, 4-methylcyclohexan-1-one (2.6 g, 19.9 mmol) and sodium cyanoborohydride (1.3 g, 19.9 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
将反应液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到780毫克白色固体4-(4-甲基环己基)哌嗪-2-酮(收率：36.0％)。LCMS：RT＝0.95min，[M+H] +＝213.15。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 780 mg of white solid 4-(4-methylcyclohexyl)piperazin-2-one was obtained (yield: 36.0%). LCMS: RT=0.95 min, [M+H] + =213.15.
步骤D：合成(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基))-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(4-Methoxycyclohexyl))-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000145
Figure PCTCN2020130361-appb-000145
将(S)-3-(4-溴苯基)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(1.0克，1.66毫摩尔)和4-(4-甲基环己基)哌嗪-2-酮(704毫克，3.22摩尔)溶于甲苯(15.0毫升)的溶液中。随后，向上述溶液中加入N，N-二甲基乙-1,2-二胺(292毫克,3.22毫摩尔)，碘化亚铜(315毫克，1.66毫摩尔)和碳酸铯(1克，3.22毫摩尔)。在110摄氏度下搅拌18个小时。Add (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazine-1- A solution of tert-butyl methyl)propionate (1.0 g, 1.66 mmol) and 4-(4-methylcyclohexyl)piperazin-2-one (704 mg, 3.22 mol) were dissolved in toluene (15.0 mL). Subsequently, to the above solution was added N,N-dimethylethyl-1,2-diamine (292 mg, 3.22 mmol), cuprous iodide (315 mg, 1.66 mmol) and cesium carbonate (1 g, 3.22 mmol). Stir at 110 degrees Celsius for 18 hours.
向反应液中加入饱和氯化铵溶液(30毫升)。混合液用乙酸乙酯(15毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/10)。得到1.1克黄色固体(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基))-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：90.0％)。LCMS：RT＝3.44min，[M+H] +＝734.33。 Saturated ammonium chloride solution (30 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (15 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/10). 1.1 g of yellow solid (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-( Tert-butyl 4-(4-(4-methoxycyclohexyl))-2-oxopiperazin-1-yl)phenyl)propionate (yield: 90.0%). LCMS: RT = 3.44 min, [M+H] + =734.33.
步骤E：合成(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪)-1-基)苯基)丙酸叔丁酯Step E: Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(4 -Methoxycyclohexyl)-2-oxopiperazin)-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000146
Figure PCTCN2020130361-appb-000146
将(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基))-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(1.1克，1.5毫摩尔)和1,3-二甲基嘧啶-2,4,6(1H,3H,5H)-三酮烷(1.6克，10.0毫摩尔)溶于二氯甲烷(10.0毫升)中。随后，向上述溶液中加入四(三苯基膦)钯(103毫克，0.06毫摩尔)。在40摄氏度下搅拌过夜。Add (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4 -(4-Methoxycyclohexyl))-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate (1.1 g, 1.5 mmol) and 1,3-dimethylpyrimidine-2 ,4,6(1H,3H,5H)-trione alkane (1.6 g, 10.0 mmol) was dissolved in dichloromethane (10.0 mL). Subsequently, tetrakis(triphenylphosphine)palladium (103 mg, 0.06 mmol) was added to the above solution. Stir overnight at 40 degrees Celsius.
向反应液中加入饱和碳酸氢钠溶液(20毫升)。混合液用乙酸乙酯(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(20毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到450毫克黄色固体(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)叔丁基酯基)苯基)丙酸叔丁酯(收率：45.0％)。LCMS：RT＝3.05min，[M+H] +＝654.30。 Saturated sodium bicarbonate solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (20 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 450 mg of yellow solid (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-iso Propyl-2-oxopiperazin-1-yl)tert-butyl esteryl)phenyl)propionate (yield: 45.0%). LCMS: RT = 3.05 min, [M+H] + =654.30.
步骤F：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step F: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000147
Figure PCTCN2020130361-appb-000147
将(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪)-1-基)苯基)丙酸叔丁酯(450毫克，0.68毫摩尔)和原甲酸三乙酯(713毫克，4.82毫摩尔)溶于醋酸(5.0毫升)中。随后，向上述溶液中加入叠氮化钠(313毫克，4.82毫摩尔)。在70摄氏度下搅拌1小时。(S)-2-(4-(2-Amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(4-methoxy (Cyclohexyl)-2-oxopiperazin)-1-yl)phenyl)propionate (450 mg, 0.68 mmol) and triethyl orthoformate (713 mg, 4.82 mmol) dissolved in acetic acid (5.0 mL). Subsequently, sodium azide (313 mg, 4.82 mmol) was added to the above solution. Stir at 70 degrees Celsius for 1 hour.
向反应液中加入饱和碳酸氢钠溶液(40毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到330毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：71.0％)。LCMS：RT＝3.05min，[M+H] +＝707.31。 Saturated sodium bicarbonate solution (40 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 330 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-butyl 3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionate (yield: 71.0%). LCMS: RT = 3.05 min, [M+H] + =707.31.
步骤G：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-(甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酸Step G: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-(methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid
Figure PCTCN2020130361-appb-000148
Figure PCTCN2020130361-appb-000148
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(330毫克，0.49毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(4-Methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (330 mg, 0.49 mmol) dissolved in dichloromethane (4.0 mL) in. Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。得到280毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-(甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酸(收率：86.0％)。The reaction solution was concentrated under reduced pressure. Obtain 280 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- 3-(4-(4-(4-(methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid (yield: 86.0%).
步骤H：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step H: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2-dicarboxy Di-tert-butyl ester
Figure PCTCN2020130361-appb-000149
Figure PCTCN2020130361-appb-000149
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-(甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酸(280毫克，0.43毫摩尔)和5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯(157毫克0.47摩尔)溶于N,N-二甲基甲酰胺(10.0毫升)的溶液中。随后，向上述溶液中加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(245毫克，0.64毫摩尔)，N,N-二异丙基乙胺(111毫克，0.86毫摩尔)，在室温下搅拌18个小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(4-(Methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid (280 mg, 0.43 mmol) and 5-amino-1H-indole-1 Di-tert-butyl 2-dicarboxylate (157 mg 0.47 mol) was dissolved in a solution of N,N-dimethylformamide (10.0 ml). Then, 2-(7-oxybenzoic acid) was added to the above solution. Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (245 mg, 0.64 mmol), N,N-diisopropylethylamine (111 mg, 0.86 mmol) , Stir at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(20毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到430毫克无色油状物(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：103.0％)。LCMS：RT＝3.57min，[M+H] +＝865.40。 Saturated ammonium chloride solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 430 mg of colorless oil (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-1,2 -Di-tert-butyl dicarboxylic acid (yield: 103.0%). LCMS: RT = 3.57 min, [M+H] + =865.40.
步骤I：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step I: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000150
Figure PCTCN2020130361-appb-000150
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(430毫克，0.44毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert Butyl ester (430 mg, 0.44 mmol) was dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。所得残余物用制备HPLC纯化得到34.2毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：10.0％)。LCMS：RT＝2.83min，[M+H] +＝809.28。 1H NMR(500MHz,DMSO)δ12.91(s,1H),11.73(s,1H),10.09(d,J＝38.0Hz,2H),9.76(s,1H),7.99(s,1H),7.94(d,J＝1.7Hz,1H),7.81(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.2Hz,1H),7.36(dt,J＝19.8,8.6Hz,6H),7.06(t,J＝4.0Hz,1H),5.38(s,1H),3.44-3.29(m,4H),3.23(t,J＝8.0Hz,4H),3.13(dt,J＝21.6,10.9Hz,2H),2.13(d,J＝13.2Hz,2H),2.04–1.80(m,3H),1.65(t,J＝51.2Hz,3H),1.44(t,J＝13.0Hz,2H),1.33–1.09(m,2H)。 The reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 34.2 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-di Oxopiperazin-1-yl)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indyl Dole-2-carboxylic acid (yield: 10.0%). LCMS: RT = 2.83 min, [M+H] + =809.28. 1 H NMR (500MHz, DMSO) δ 12.91 (s, 1H), 11.73 (s, 1H), 10.09 (d, J = 38.0 Hz, 2H), 9.76 (s, 1H), 7.99 (s, 1H), 7.94(d,J=1.7Hz,1H),7.81(d,J=8.6Hz,1H),7.77(dd,J=8.6,2.2Hz,1H),7.36(dt,J=19.8,8.6Hz,6H ), 7.06(t,J=4.0Hz,1H),5.38(s,1H),3.44-3.29(m,4H),3.23(t,J=8.0Hz,4H),3.13(dt,J=21.6, 10.9Hz, 2H), 2.13 (d, J = 13.2Hz, 2H), 2.04-1.80 (m, 3H), 1.65 (t, J = 51.2Hz, 3H), 1.44 (t, J = 13.0Hz, 2H) ,1.33-1.09(m,2H).
实施例29Example 29
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000151
Figure PCTCN2020130361-appb-000151
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(2-甲氧基乙基)哌嗪-2-酮Step A: Synthesis of 4-(2-methoxyethyl)piperazin-2-one
Figure PCTCN2020130361-appb-000152
Figure PCTCN2020130361-appb-000152
将哌嗪-2-酮(2.0克，20.0毫摩尔)溶于乙腈(20.0毫升)中。随后，向上述溶液中加入1-溴-2-甲氧基乙烷(5.6克，40.0毫摩尔)和碳酸钾(5.5克，40.0毫摩尔)。在100摄氏度下搅拌过夜。Piperazine-2-one (2.0 g, 20.0 mmol) was dissolved in acetonitrile (20.0 mL). Subsequently, 1-bromo-2-methoxyethane (5.6 g, 40.0 mmol) and potassium carbonate (5.5 g, 40.0 mmol) were added to the above solution. Stir overnight at 100 degrees Celsius.
将反应液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到800毫克白色固体4-(2-甲氧基乙基)哌嗪-2-酮(收率：25.0％)。LCMS：RT＝0.90min，[M+H] +＝159.15。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 800 mg of white solid 4-(2-methoxyethyl)piperazin-2-one was obtained (yield: 25.0%). LCMS: RT=0.90 min, [M+H] + =159.15.
步骤B：合成(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-)2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step B: Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(2-Methoxyethyl)-)2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000153
Figure PCTCN2020130361-appb-000153
将(S)-3-(4-溴苯基)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(1.5克，2.5毫摩尔)和4-(2-甲氧基乙基)哌嗪-2-酮(767毫克，4.98毫摩尔)溶于甲苯(15.0毫升)的溶液中。随后，向上述溶液中加入N，N-二甲基乙-1,2-二胺(438毫克,4.98毫摩尔)，碘化亚铜(473毫克，2.50毫摩尔)和碳酸铯(1.62克，4.98毫摩尔)。在110摄氏度下搅拌18个小时。Add (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazine-1- A solution of tert-butyl propionate (1.5 g, 2.5 mmol) and 4-(2-methoxyethyl)piperazin-2-one (767 mg, 4.98 mmol) in toluene (15.0 mL) in. Subsequently, N,N-dimethylethyl-1,2-diamine (438 mg, 4.98 mmol), cuprous iodide (473 mg, 2.50 mmol) and cesium carbonate (1.62 g, 4.98 mmol). Stir at 110 degrees Celsius for 18 hours.
向反应液中加入饱和氯化铵溶液(30毫升)。混合液用乙酸乙酯(15毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/10)。得到700毫克黄色固体(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-)2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：48.0％)。LCMS：RT＝3.36min，[M+H] +＝680.31。 Saturated ammonium chloride solution (30 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (15 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/10). Obtain 700 mg of yellow solid (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-( Tert-Butyl 4-(4-(2-methoxyethyl)-)2-oxopiperazin-1-yl)phenyl)propionate (yield: 48.0%). LCMS: RT = 3.36 min, [M+H] + =680.31.
步骤C：合成(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪)-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(2 -Methoxyethyl)-2-oxopiperazin)-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000154
Figure PCTCN2020130361-appb-000154
将(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-)2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(700毫克，1.0毫摩尔)和1,3-二甲基嘧啶-2,4,6(1H,3H,5H)-三酮烷(964毫克，6.0毫摩尔)溶于二氯甲烷(10.0毫升)中。随后，向上述溶液中加入四(三苯基膦)钯(47毫克，0.04毫摩尔)。在40摄氏度下搅拌过夜。Add (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4 -(2-Methoxyethyl)-)2-oxopiperazin-1-yl)phenyl)tert-butyl propionate (700 mg, 1.0 mmol) and 1,3-dimethylpyrimidine-2 ,4,6(1H,3H,5H)-trione alkane (964 mg, 6.0 mmol) was dissolved in dichloromethane (10.0 mL). Subsequently, tetrakis(triphenylphosphine)palladium (47 mg, 0.04 mmol) was added to the above solution. Stir overnight at 40 degrees Celsius.
向反应液中加入饱和碳酸氢钠溶液(20毫升)。混合液用乙酸乙酯(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(20毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到500毫克黄色固体(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪)-1-基)苯基)丙酸叔丁酯(收率：80.0％)。LCMS：RT＝2.95min，[M+H] +＝600.15。 Saturated sodium bicarbonate solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (20 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 500 mg of yellow solid (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-( 2-Methoxyethyl)-2-oxopiperazin)-1-yl)phenyl)propionic acid tert-butyl ester (yield: 80.0%). LCMS: RT = 2.95 min, [M+H] + =600.15.
步骤D：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000155
Figure PCTCN2020130361-appb-000155
将(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪)-1-基)苯基)丙酸叔丁酯(500毫克，0.83毫摩尔)和原甲酸三乙酯(617毫克，4.17毫摩尔)溶于醋酸(5.0毫升)中。随后，向上述溶液中加入叠氮化钠(270毫克，4.17毫摩尔)。在70摄氏度下搅拌1小时。(S)-2-(4-(2-Amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(2-methoxy (Ethylethyl)-2-oxopiperazin)-1-yl)phenyl)propionate (500 mg, 0.83 mmol) and triethyl orthoformate (617 mg, 4.17 mmol) dissolved in acetic acid (5.0 mL). Subsequently, sodium azide (270 mg, 4.17 mmol) was added to the above solution. Stir at 70 degrees Celsius for 1 hour.
向反应液中加入饱和碳酸氢钠溶液(40毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到420毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：77.0％)。LCMS：RT＝2.95min，[M+H] +＝653.25。 Saturated sodium bicarbonate solution (40 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 420 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-butyl 3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionate (yield: 77.0%). LCMS: RT = 2.95 min, [M+H] + =653.25.
步骤E：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-(甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酸Step E: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(2-(Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionic acid
Figure PCTCN2020130361-appb-000156
Figure PCTCN2020130361-appb-000156
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(200毫克，0.4毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (200 mg, 0.4 mmol) dissolved in dichloromethane (4.0 mL) in. Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。得到180毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-(甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酸(收率：95.0％)。The reaction solution was concentrated under reduced pressure. Obtain 180 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- 3-(4-(4-(2-(methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionic acid (yield: 95.0%).
步骤F：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step F: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-1,2-dicarboxy Di-tert-butyl ester
Figure PCTCN2020130361-appb-000157
Figure PCTCN2020130361-appb-000157
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-(甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酸(180毫克，0.30毫摩尔)和5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯(361毫克0.36摩尔)溶于N,N-二甲基甲酰胺(10.0毫升)的溶液中。随后，向上述溶液中加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(171毫克，0.45毫摩尔)，N,N-二异丙基乙胺(77毫克，0.60毫摩尔)，在室温下搅拌18个小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(2-(Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionic acid (180 mg, 0.30 mmol) and 5-amino-1H-indole-1 Di-tert-butyl 2-dicarboxylate (361 mg 0.36 mol) was dissolved in a solution of N,N-dimethylformamide (10.0 ml). Subsequently, 2-(7-oxybenzoic acid) was added to the above solution. Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (171 mg, 0.45 mmol), N,N-diisopropylethylamine (77 mg, 0.60 mmol) , Stir at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(20毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到17毫克无色油状物(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：6.1％)。LCMS：RT＝3.49min，[M+H] +＝810.35。 Saturated ammonium chloride solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtained 17 mg of colorless oil (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-1,2 -Di-tert-butyl dicarboxylic acid (yield: 6.1%). LCMS: RT = 3.49 min, [M+H] + =810.35.
步骤G：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step G: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000158
Figure PCTCN2020130361-appb-000158
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(170毫克，0.018毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl ester (170 mg, 0.018 mmol) was dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。所得残余物用制备HPLC纯化得到12毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-甲氧基乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：81.0％)。LCMS：RT＝2.73min，[M+H] +＝755.24。 The reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 12 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-di Oxopiperazin-1-yl)-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indyl Dole-2-carboxylic acid (yield: 81.0%). LCMS: RT = 2.73 min, [M+H] + =755.24.
实施例30Example 30
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸化合物Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid compound
Figure PCTCN2020130361-appb-000159
Figure PCTCN2020130361-appb-000159
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(四氢-2H-吡喃-4-基)哌嗪-2-酮Step A: Synthesis of 4-(tetrahydro-2H-pyran-4-yl)piperazin-2-one
Figure PCTCN2020130361-appb-000160
Figure PCTCN2020130361-appb-000160
将哌嗪-2-酮(500毫克，2.5毫摩尔)溶于甲醇(10.0毫升)和乙酸(0.2毫升)的混合溶液中。随后，向上述溶液中加入四氢-4H-吡喃-4-酮(788毫克，5.0毫摩尔)和氰基硼氢化钠(317毫克，5.0毫摩尔)。在60摄氏度下搅拌过夜。Piperazine-2-one (500 mg, 2.5 mmol) was dissolved in a mixed solution of methanol (10.0 mL) and acetic acid (0.2 mL). Subsequently, tetrahydro-4H-pyran-4-one (788 mg, 5.0 mmol) and sodium cyanoborohydride (317 mg, 5.0 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
将反应液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到420毫克白色固体4-(4-甲基环己基)哌嗪-2-酮(收率：69.0％)。LCMS：RT＝0.96min，[M+H] +＝185.12。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 420 mg of white solid 4-(4-methylcyclohexyl)piperazin-2-one was obtained (yield: 69.0%). LCMS: RT=0.96 min, [M+H] + =185.12.
步骤B：合成(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸叔丁酯Step B: Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000161
Figure PCTCN2020130361-appb-000161
将(S)-3-(4-溴苯基)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(1.0克，1.6毫摩尔)和4-(四氢-2H-吡喃-4-基)哌嗪-2-酮(611毫克，3.3毫摩尔)溶于甲苯(15.0毫升)的溶液中。随后，向上述溶液中加入N，N-二甲基乙-1,2-二胺(292毫克,3.3毫摩尔)，碘化亚铜(315毫克，1.6毫摩尔)和碳酸铯(1.08克，3.3毫摩尔)。在110摄氏度下搅拌18个小时。Add (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazine-1- Yl) tert-butyl propionate (1.0 g, 1.6 mmol) and 4-(tetrahydro-2H-pyran-4-yl)piperazin-2-one (611 mg, 3.3 mmol) dissolved in toluene (15.0 Ml) of the solution. Subsequently, N,N-dimethylethyl-1,2-diamine (292 mg, 3.3 mmol), cuprous iodide (315 mg, 1.6 mmol) and cesium carbonate (1.08 g, 3.3 mmol). Stir at 110 degrees Celsius for 18 hours.
向反应液中加入饱和氯化铵溶液(30毫升)。混合液用乙酸乙酯(15毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/10)。得到590毫克黄色固体(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸叔丁酯(收率：50％)。LCMS：RT＝3.31min，[M+H] +＝706.33。 Saturated ammonium chloride solution (30 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (15 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/10). Obtain 590 mg of yellow solid (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-( Tert-Butyl 4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionate (yield: 50%). LCMS: RT = 3.31 min, [M+H] + =706.33.
步骤C：合成(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2-oxo -4-(Tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000162
Figure PCTCN2020130361-appb-000162
将(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸叔丁酯(590毫克，0.83毫摩尔)和1,3-二甲基嘧啶-2,4,6(1H,3H,5H)-三酮烷(913毫克，5.00毫摩尔)溶于二氯甲烷(10.0毫升)中。随后，向上述溶液中加入四(三苯基膦)钯(57毫克，0.05毫摩尔)。在40摄氏度下搅拌过夜。Add (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2 -Oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl) tert-butyl propionate (590 mg, 0.83 mmol) and 1,3-dimethyl Pyrimidine-2,4,6(1H,3H,5H)-trione alkane (913 mg, 5.00 mmol) was dissolved in dichloromethane (10.0 mL). Subsequently, tetrakis(triphenylphosphine)palladium (57 mg, 0.05 mmol) was added to the above solution. Stir overnight at 40 degrees Celsius.
向反应液中加入饱和碳酸氢钠溶液(20毫升)。混合液用乙酸乙酯(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(20毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到450毫克黄色固体(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸叔丁酯(收率：86.0％)。LCMS：RT＝2.93min，[M+H] +＝626.30。 Saturated sodium bicarbonate solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (20 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 450 mg of yellow solid (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2-oxygen Tert-butyl 4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionate (yield: 86.0%). LCMS: RT = 2.93 min, [M+H] + =626.30.
步骤D：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000163
Figure PCTCN2020130361-appb-000163
将(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸叔丁酯(450毫克，0.71毫摩尔)和原甲酸三乙酯(840毫克，5.00毫摩尔)溶于醋酸(5.0毫升)中。随后，向上述溶液中加入叠氮化钠(327毫克，5.00毫摩尔)。在70摄氏度下搅拌1小时。(S)-2-(4-(2-Amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2-oxo-4- (Tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)tert-butyl propionate (450 mg, 0.71 mmol) and triethyl orthoformate (840 mg, 5.00 mmol) Dissolve in acetic acid (5.0 ml). Subsequently, sodium azide (327 mg, 5.00 mmol) was added to the above solution. Stir at 70 degrees Celsius for 1 hour.
向反应液中加入饱和碳酸氢钠溶液(40毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到400毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸叔丁酯(收率：86.0％)。LCMS：RT＝2.92min，[M+H] +＝679.27。 Saturated sodium bicarbonate solution (40 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 400 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-butyl 3-(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionate (yield: 86.0%). LCMS: RT = 2.92 min, [M+H] + =679.27.
步骤E：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸Step E: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionic acid
Figure PCTCN2020130361-appb-000164
Figure PCTCN2020130361-appb-000164
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸叔丁酯(400毫克，0.6毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl) tert-butyl propionate (400 mg, 0.6 mmol) dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。得到360毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸收率：93.0％)。The reaction solution was concentrated under reduced pressure. Obtain 360 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- 3-(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionic acid yield: 93.0%).
步骤F：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step F: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-1, Di-tert-butyl 2-dicarboxylate
Figure PCTCN2020130361-appb-000165
Figure PCTCN2020130361-appb-000165
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酸(360毫克，0.55毫摩尔)和5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯(202毫克0.60摩尔)溶于N,N-二甲基甲酰胺(10.0 毫升)的溶液中。随后，向上述溶液中加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(315毫克，0.82毫摩尔)，N,N-二异丙基乙胺(142毫克，1.10毫摩尔)，在室温下搅拌18个小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionic acid (360 mg, 0.55 mmol) and 5-amino-1H-indole Dole-1,2-dicarboxylic acid di-tert-butyl ester (202 mg 0.60 mol) was dissolved in a solution of N,N-dimethylformamide (10.0 ml). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (315 mg, 0.82 mmol) was added to the above solution, N,N -Diisopropylethylamine (142 mg, 1.10 mmol), stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(20毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到380毫克无色油状物(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：71％)。LCMS：RT＝3.47min，[M+H] +＝837.37。 Saturated ammonium chloride solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 380 mg of colorless oil (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole -Di-tert-butyl 1,2-dicarboxylic acid (yield: 71%). LCMS: RT = 3.47 min, [M+H] + =837.37.
步骤G：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step G: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2- carboxylic acid
Figure PCTCN2020130361-appb-000166
Figure PCTCN2020130361-appb-000166
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(380毫克，0.4毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate (380 mg, 0.4 mmol) was dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。所得残余物用制备HPLC纯化得到52毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：19.0％)。LCMS：RT＝2.74min，[M+H] +＝781.25。 1H NMR(500MHz,DMSO)δ12.91(s,1H),11.72(s,1H),10.13(s,1H),9.75(s,1H),7.99(s,1H),7.94(d,J＝2.0Hz,1H),7.81(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.2Hz,1H),7.42–7.30(m,5H),7.07(d,J＝1.7Hz,1H),5.38(s,1H),3.98(d,J＝9.5Hz,3H),3.35–3.29(m,3H),3.13(dd,J＝14.4,10.2Hz,2H),2.94(s,3H),2.00(d,J＝7.5Hz,3H),1.65(s,2H),1.24(s,2H)。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC to obtain 52 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-di Oxopiperazin-1-yl)-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionamido) -1H-indole-2-carboxylic acid (yield: 19.0%). LCMS: RT = 2.74 min, [M+H] + =781.25. 1 H NMR (500MHz, DMSO) δ 12.91 (s, 1H), 11.72 (s, 1H), 10.13 (s, 1H), 9.75 (s, 1H), 7.99 (s, 1H), 7.94 (d, J =2.0Hz,1H),7.81(d,J=8.6Hz,1H),7.77(dd,J=8.6,2.2Hz,1H),7.42-7.30(m,5H),7.07(d,J=1.7Hz ,1H),5.38(s,1H),3.98(d,J＝9.5Hz,3H),3.35-3.29(m,3H),3.13(dd,J＝14.4,10.2Hz,2H),2.94(s, 3H), 2.00 (d, J=7.5 Hz, 3H), 1.65 (s, 2H), 1.24 (s, 2H).
实施例31Example 31
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-4-基)哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-4-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000167
Figure PCTCN2020130361-appb-000167
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(4-羟基环己基)哌嗪-2-酮Step A: Synthesis of 4-(4-hydroxycyclohexyl)piperazin-2-one
Figure PCTCN2020130361-appb-000168
Figure PCTCN2020130361-appb-000168
将哌嗪-2-酮(1.5克，7.5毫摩尔)溶于甲醇(20.0毫升)和乙酸(0.4毫升)的混合溶液中。随后，向上述溶液中加入4-羟基环己烷-1-酮(1.7克，15.0毫摩尔)和氰基硼氢化钠(951毫克，15.0毫摩尔)。在60摄氏度下搅拌过夜。Piperazine-2-one (1.5 g, 7.5 mmol) was dissolved in a mixed solution of methanol (20.0 mL) and acetic acid (0.4 mL). Subsequently, 4-hydroxycyclohexane-1-one (1.7 g, 15.0 mmol) and sodium cyanoborohydride (951 mg, 15.0 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
将反应液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到750毫克白色固体4-(4-甲基环己基)哌嗪-2-酮(收率：50.0％)。LCMS：RT＝0.82，[M+H] +＝199.14。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 750 mg of white solid 4-(4-methylcyclohexyl)piperazin-2-one was obtained (yield: 50.0%). LCMS: RT=0.82, [M+H] + =199.14.
步骤B：合成(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基))-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step B: Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(4-Hydroxycyclohexyl))-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000169
Figure PCTCN2020130361-appb-000169
将(S)-3-(4-溴苯基)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(600毫克，0.99毫摩尔)和4-(4-羟基环己基)哌嗪-2-酮(394毫克，1.99毫摩尔)溶于甲苯(15毫升)的溶液中。随后，向上述溶液中加入N，N-二甲基乙-1,2-二胺(175毫克,1.99毫摩尔)，碘化亚铜(189毫克，0.99毫摩尔)和碳酸铯(645毫克，1.99毫摩尔)。在110摄氏度下搅拌18个小时。Add (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazine-1- A solution of tert-butyl methyl)propionate (600 mg, 0.99 mmol) and 4-(4-hydroxycyclohexyl)piperazin-2-one (394 mg, 1.99 mmol) were dissolved in toluene (15 mL). Subsequently, to the above solution was added N,N-dimethylethane-1,2-diamine (175 mg, 1.99 mmol), cuprous iodide (189 mg, 0.99 mmol) and cesium carbonate (645 mg, 1.99 mmol). Stir at 110 degrees Celsius for 18 hours.
向反应液中加入饱和氯化铵溶液(30毫升)。混合液用乙酸乙酯(15毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/10)。得到460毫克黄色固体(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基))-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：64.0％)。LCMS：RT＝3.10min，[M+H] +＝720.34。 Saturated ammonium chloride solution (30 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (15 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/10). Obtain 460 mg of yellow solid (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-( Tert-butyl 4-(4-(4-hydroxycyclohexyl))-2-oxopiperazin-1-yl)phenyl)propionate (yield: 64.0%). LCMS: RT = 3.10 min, [M+H] + =720.34.
步骤C：合成(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪)-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(4 -Hydroxycyclohexyl)-2-oxopiperazin)-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000170
Figure PCTCN2020130361-appb-000170
将(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基))-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(550毫克，0.76毫摩尔)和1,3-二甲基嘧啶-2,4,6(1H,3H,5H)-三酮烷(834毫克，5.30毫摩尔)溶于二氯甲烷(10.0毫升)中。随后，向上述溶液中加入四(三苯基膦)钯(52毫克，0.045毫摩尔)。在40摄氏度下搅拌过夜。Add (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4 -(4-Hydroxycyclohexyl))-2-oxopiperazin-1-yl)phenyl)tert-butyl propionate (550 mg, 0.76 mmol) and 1,3-dimethylpyrimidine-2,4 ,6(1H,3H,5H)-trione alkane (834 mg, 5.30 mmol) was dissolved in dichloromethane (10.0 mL). Subsequently, tetrakis(triphenylphosphine)palladium (52 mg, 0.045 mmol) was added to the above solution. Stir overnight at 40 degrees Celsius.
向反应液中加入饱和碳酸氢钠溶液(20毫升)。混合液用乙酸乙酯(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(20毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到460毫克黄色固体(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪)-1-基)苯基)丙酸叔丁酯(收率：94.0％)。LCMS：RT＝2.84min，[M+H] +＝640.28。 Saturated sodium bicarbonate solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (20 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 460 mg of yellow solid (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-( Tert-butyl 4-hydroxycyclohexyl)-2-oxopiperazine)-1-yl)phenyl)propionate (yield: 94.0%). LCMS: RT = 2.84 min, [M+H] + = 640.28.
步骤D：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000171
Figure PCTCN2020130361-appb-000171
将(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪)-1-基)苯基)丙酸叔丁酯(450毫克，0.7毫摩尔)和原甲酸三乙酯(1000毫克，7.0毫摩尔)溶于醋酸(5.0毫升)中。随后，向上述溶液中加入叠氮化钠(456毫克，7.0毫摩尔)。在70摄氏度下搅拌1小时。(S)-2-(4-(2-Amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(4-(4-hydroxyl ring (Hexyl)-2-oxopiperazin)-1-yl)phenyl)tert-butyl propionate (450 mg, 0.7 mmol) and triethyl orthoformate (1000 mg, 7.0 mmol) dissolved in acetic acid (5.0 Milliliters). Subsequently, sodium azide (456 mg, 7.0 mmol) was added to the above solution. Stir at 70 degrees Celsius for 1 hour.
向反应液中加入饱和碳酸氢钠溶液(40毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到110毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(收率：22.0％)。LCMS：RT＝2.83min，[M+H] +＝693.28。 Saturated sodium bicarbonate solution (40 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 110 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-butyl 3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionate (yield: 22.0%). LCMS: RT = 2.83 min, [M+H] + =693.28.
步骤E：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酸Step E: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid
Figure PCTCN2020130361-appb-000172
Figure PCTCN2020130361-appb-000172
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酸叔丁酯(110毫克，0.15毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (110 mg, 0.15 mmol) was dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。得93毫克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酸(收率：92.0％)。The reaction solution was concentrated under reduced pressure. Obtain 93 mg of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- 3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid (yield: 92.0%).
步骤F：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-2-羧酸叔丁酯Step F: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionylamino)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000173
Figure PCTCN2020130361-appb-000173
将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酸(83毫克，0.13毫摩尔)和5-氨基-1H-吲哚-2-羧酸叔丁酯(36毫克0.15摩尔)溶于N,N-二甲基甲酰胺(10.0毫升)的溶液中。随后，向上述溶液中加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(74毫克，0.19毫摩尔)，N,N-二异丙基乙胺(33毫克，0.26毫摩尔)，在室温下搅拌18个小时。Add (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid (83 mg, 0.13 mmol) and 5-amino-1H-indole-2-carboxylic acid Tert-butyl ester (36 mg 0.15 mol) was dissolved in a solution of N,N-dimethylformamide (10.0 ml). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (74 mg, 0.19 mmol) was added to the above solution, N, N -Diisopropylethylamine (33 mg, 0.26 mmol), stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(20毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到65毫克无色油状物(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-2-羧酸叔丁酯(收率：59.0％)。LCMS：RT＝3.13min，[M+H] +＝851.33。 Saturated ammonium chloride solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 65 mg of colorless oil (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid tert Butyl ester (yield: 59.0%). LCMS: RT = 3.13 min, [M+H] + =851.33.
步骤G：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸化合物Step G: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid compound
Figure PCTCN2020130361-appb-000174
Figure PCTCN2020130361-appb-000174
将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-2-羧酸叔丁酯(65毫克，0.07毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。(S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (65 mg, 0.07 mmol) was dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。所得残余物用制备HPLC纯化得到50毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸化合物(收率：83.0％)。LCMS：RT＝2.33min，[M+H] +＝795.27。 1H NMR(400MHz,DMSO)δ11.74(s,1H),10.14(s,1H),9.77(s,1H),8.00(s,1H),7.94(d,J＝1.8Hz,1H),7.82(d,J＝8.6Hz,1H),7.78(dd,J＝8.7,2.2Hz,1H),7.42–7.30(m,6H),7.08(d,J＝1.9Hz,1H),5.44-5.36(m,1H),4.12-3.13(m,13H),3.13(dd,J＝14.8,10.4Hz,1H),2.07-2.00(m,1H),1.96-1.91(m,1H),1.86-1.75(m,3H),1.61-1.41(m,3H)。 The reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 50 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-di Oxopiperazin-1-yl)-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole- 2-carboxylic acid compound (yield: 83.0%). LCMS: RT = 2.33 min, [M+H] + =795.27. 1 H NMR (400MHz, DMSO) δ 11.74 (s, 1H), 10.14 (s, 1H), 9.77 (s, 1H), 8.00 (s, 1H), 7.94 (d, J = 1.8 Hz, 1H), 7.82(d,J=8.6Hz,1H),7.78(dd,J=8.7,2.2Hz,1H),7.42-7.30(m,6H),7.08(d,J=1.9Hz,1H),5.44-5.36 (m,1H),4.12-3.13(m,13H),3.13(dd,J=14.8,10.4Hz,1H),2.07-2.00(m,1H),1.96-1.91(m,1H),1.86-1.75 (m, 3H), 1.61-1.41 (m, 3H).
实施例32Example 32
合成5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000175
Figure PCTCN2020130361-appb-000175
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(四氢-2H-吡喃-3-基)哌嗪-2-酮Step A: Synthesis of 4-(tetrahydro-2H-pyran-3-yl)piperazin-2-one
Figure PCTCN2020130361-appb-000176
Figure PCTCN2020130361-appb-000176
将哌嗪-2-酮(500毫克，4.9毫摩尔)溶于甲醇(10.0毫升)和乙酸(0.2毫升)的混合溶液中。随后，向上述溶液中加入二氢-2H-吡喃-3(4H)-酮(1.0克，10.0毫摩尔)和氰基硼氢化钠(627毫克，10.0毫摩尔)。在60摄氏度下搅拌过夜。Piperazine-2-one (500 mg, 4.9 mmol) was dissolved in a mixed solution of methanol (10.0 mL) and acetic acid (0.2 mL). Subsequently, dihydro-2H-pyran-3(4H)-one (1.0 g, 10.0 mmol) and sodium cyanoborohydride (627 mg, 10.0 mmol) were added to the above solution. Stir overnight at 60 degrees Celsius.
将反应液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到400毫克白色固体4-(四氢-2H-吡喃-3-基)哌嗪-2-酮(收率：43.0％)。LCMS：RT＝1.04min，[M+H] +＝185.12。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 400 mg of white solid 4-(tetrahydro-2H-pyran-3-yl)piperazin-2-one was obtained (yield: 43.0%). LCMS: RT=1.04 min, [M+H] + =185.12.
步骤B：合成(2S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢)-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸叔丁酯Step B: Synthesis of (2S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(2-oxo-4-(tetrahydro)-2H-pyran-3-yl)piperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000177
Figure PCTCN2020130361-appb-000177
将(S)-3-(4-溴苯基)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(600毫克，0.99毫摩尔)和4-(四氢-2H-吡喃-3-基)哌嗪-2-酮(370毫克，1.99毫摩尔)溶于甲苯(15.0毫升)的溶液中。随后，向上述溶液中加入N，N-二甲基乙-1,2-二胺(175毫克,1.99毫摩尔)，碘化亚铜(189毫克，0.99毫摩尔)和碳酸铯(645毫克，1.99毫摩尔)。在110摄氏度下搅拌18个小时。Add (S)-3-(4-bromophenyl)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazine-1- Yl) tert-butyl propionate (600 mg, 0.99 mmol) and 4-(tetrahydro-2H-pyran-3-yl)piperazin-2-one (370 mg, 1.99 mmol) dissolved in toluene (15.0 Ml) of the solution. Subsequently, to the above solution was added N,N-dimethylethane-1,2-diamine (175 mg, 1.99 mmol), cuprous iodide (189 mg, 0.99 mmol) and cesium carbonate (645 mg, 1.99 mmol). Stir at 110 degrees Celsius for 18 hours.
向反应液中加入饱和氯化铵溶液(30毫升)，混合液用乙酸乙酯(15毫升×3次)萃取，合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/石油醚＝1/10)。得到370毫克黄色固体(2S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢)-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸叔丁酯(收率：52.0％)。LCMS：RT＝3.48min，[M+H] +＝706.33。 Saturated ammonium chloride solution (30 mL) was added to the reaction solution, the mixture was extracted with ethyl acetate (15 mL×3 times), and the organic phases were combined. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/10). Obtain 370 mg of yellow solid (2S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-( Tert-Butyl 4-(2-oxo-4-(tetrahydro)-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionate (yield: 52.0%). LCMS: RT = 3.48 min, [M+H] + =706.33.
步骤C：合成(2S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (2S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2-oxo -4-(Tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000178
Figure PCTCN2020130361-appb-000178
将(2S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢)-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸叔丁酯(370毫克，0.52毫摩尔)和1,3-二甲基嘧啶-2,4,6(1H,3H,5H)-三酮烷(572毫克，3.6毫摩尔)溶于二氯甲烷(10.0毫升)中。随后，向上述溶液中加入四(三苯基膦)钯(36毫克，0.031毫摩尔)。在40摄氏度下搅拌过夜。Add (2S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2 -Oxo-4-(tetrahydro)-2H-pyran-3-yl)piperazin-1-yl)phenyl)tert-butyl propionate (370 mg, 0.52 mmol) and 1,3-dimethyl Pyrimidine-2,4,6(1H,3H,5H)-trione alkane (572 mg, 3.6 mmol) was dissolved in dichloromethane (10.0 mL). Subsequently, tetrakis(triphenylphosphine)palladium (36 mg, 0.031 mmol) was added to the above solution. Stir overnight at 40 degrees Celsius.
向反应液中加入饱和碳酸氢钠溶液(20毫升)。混合液用乙酸乙酯(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(20毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到260毫克黄色固体(2S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸叔丁酯(收率：79.0％)。LCMS：RT＝3.06min，[M+H] +＝626.27。 Saturated sodium bicarbonate solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (20 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 260 mg of yellow solid (2S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2-oxygen Tert-butyl 4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionate (yield: 79.0%). LCMS: RT = 3.06 min, [M+H] + =626.27.
步骤D：合成(2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000179
Figure PCTCN2020130361-appb-000179
将(2S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸叔丁酯(260毫克，0.4毫摩尔)和原甲酸三乙酯(430毫克，2.9毫摩尔)溶于醋酸(5.0毫升)中。随后，向上述溶液中加入叠氮化钠(188毫克，2.9毫摩尔)。在70摄氏度下搅拌1小时。(2S)-2-(4-(2-Amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-(2-oxo-4- (Tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)tert-butyl propionate (260 mg, 0.4 mmol) and triethyl orthoformate (430 mg, 2.9 mmol) Dissolve in acetic acid (5.0 ml). Subsequently, sodium azide (188 mg, 2.9 mmol) was added to the above solution. Stir at 70 degrees Celsius for 1 hour.
向反应液中加入饱和碳酸氢钠溶液(40毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到80毫克黄色固体(2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸叔丁酯(收率：28.0％)。LCMS：RT＝3.05min，[M+H] +＝679.27。 Saturated sodium bicarbonate solution (40 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 80 mg of yellow solid (2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-butyl 3-(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionate (yield: 28.0%). LCMS: RT=3.05 min, [M+H] + =679.27.
步骤E：合成(2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸Step E: Synthesis of (2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionic acid
Figure PCTCN2020130361-appb-000180
Figure PCTCN2020130361-appb-000180
将(2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸叔丁酯(80毫克，0.11毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。Add (2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl) tert-butyl propionate (80 mg, 0.11 mmol) dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。得65毫克黄色固体(2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸(收率：89.0％)。The reaction solution was concentrated under reduced pressure. Get 65 mg of yellow solid (2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- 3-(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionic acid (yield: 89.0%).
步骤F：合成5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-2-羧酸叔丁酯Step F: Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2- Tert-butyl carboxylate
Figure PCTCN2020130361-appb-000181
Figure PCTCN2020130361-appb-000181
将(2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酸(65毫克，0.10毫摩尔)和5-氨基-1H-吲哚-2-羧酸叔丁酯(41毫克0.12摩尔)溶于N,N-二甲基甲酰胺(10.0毫升)的溶液中。随后，向上述溶液中加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(59毫克，0.15毫摩尔)，N,N-二异丙基乙胺(27毫克，0.20摩尔)，在室温下搅拌18个小时。Add (2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionic acid (65 mg, 0.10 mmol) and 5-amino-1H-indole Tert-butyl indole-2-carboxylate (41 mg 0.12 mol) was dissolved in a solution of N,N-dimethylformamide (10.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (59 mg, 0.15 mmol) was added to the above solution, N,N -Diisopropylethylamine (27 mg, 0.20 mol), stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(20毫升)。混合液用乙酸乙酯(10毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(10毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到25毫克无色油状物5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-2-羧酸叔丁酯(收率：25.7％)。LCMS：RT＝3.22min，[M+H] +＝837.32。 Saturated ammonium chloride solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (10 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (10 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtained 25 mg of colorless oil 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole Tert-Butyl-2-carboxylate (yield: 25.7%). LCMS: RT=3.22 min, [M+H] + =837.32.
步骤G：合成5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step G: Synthesis of 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2- carboxylic acid
Figure PCTCN2020130361-appb-000182
Figure PCTCN2020130361-appb-000182
将5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酰氨基)-1H-吲哚-2-羧酸叔丁酯(25毫克，0.026毫摩尔)溶于二氯甲烷(4.0毫升)中。随后，向上述溶液中加入三氟乙酸(1.0毫升)。在室温下搅拌2小时。Put 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid tert Butyl ester (25 mg, 0.026 mmol) was dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。所得残余物用制备HPLC纯化得到15毫克白色固体5-((2S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(2-氧代-4-(四氢-2H-吡喃-3-基)哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：65.0％)。LCMS：RT＝2.83min，[M+H] +＝781.25。 1H NMR(500MHz,DMSO)δ12.94(s,1H),11.74(s,1H),10.15(s,1H),9.76(s,1H),8.00(s,1H),7.94(d,J＝2.1Hz,1H),7.81(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.2Hz,1H),7.41–7.27(m,6H),7.07(d,J＝1.7Hz,1H),5.38(brs,1H),3.97-3.13(m,9H),3.13(dd,J＝14.3,10.1Hz,1H),2.54(s,2H),2.08(m,2H),1.99(m,2H),1.76(m,2H),1.60–1.38(m,3H)。 The reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 15 mg of white solid 5-((2S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-di Oxopiperazin-1-yl)-3-(4-(2-oxo-4-(tetrahydro-2H-pyran-3-yl)piperazin-1-yl)phenyl)propionamido) -1H-indole-2-carboxylic acid (yield: 65.0%). LCMS: RT = 2.83 min, [M+H] + =781.25. 1 H NMR (500MHz, DMSO) δ 12.94 (s, 1H), 11.74 (s, 1H), 10.15 (s, 1H), 9.76 (s, 1H), 8.00 (s, 1H), 7.94 (d, J ＝2.1Hz,1H),7.81(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.2Hz,1H),7.41-7.27(m,6H),7.07(d,J＝1.7Hz ,1H),5.38(brs,1H),3.97-3.13(m,9H),3.13(dd,J=14.3,10.1Hz,1H),2.54(s,2H),2.08(m,2H),1.99( m, 2H), 1.76 (m, 2H), 1.60–1.38 (m, 3H).
实施例33Example 33
合成(S)-6-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-6-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000183
Figure PCTCN2020130361-appb-000183
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成6-硝基-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of 6-nitro-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000184
Figure PCTCN2020130361-appb-000184
N2保护、冰浴下，向含有6-硝基-1H-吲哚-2-羧酸(200毫克，0.97毫摩尔)的醋酸叔丁酯(25.0毫升)中滴加70％的高氯酸(147毫克，1.46毫摩尔)，滴毕，室温反应2小时。Under N2 protection and ice bath, to tert-butyl acetate (25.0 ml) containing 6-nitro-1H-indole-2-carboxylic acid (200 mg, 0.97 mmol) was added dropwise 70% perchloric acid ( 147 mg, 1.46 mmol), after dripping, react at room temperature for 2 hours.
反应结束，加乙酸乙酯稀释，用饱和碳酸钠调pH至9，分液，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝5/1)。得到200毫克黄色油状物6-硝基-1H-吲哚-2-羧酸叔丁酯(收率：74.0％)。LCMS:RT＝4.20min,[M-H] -＝261.13。 After the reaction is over, add ethyl acetate to dilute, adjust the pH to 9 with saturated sodium carbonate, separate the layers, extract the mixture with ethyl acetate (20 ml×3 times), combine the organic phases, and use saturated brine (10 ml) for the organic phase. ×2 times) Wash, then dry with anhydrous sodium sulfate, and finally concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=5/1). 200 mg of yellow oily tert-butyl 6-nitro-1H-indole-2-carboxylate was obtained (yield: 74.0%). LCMS: RT = 4.20 min, [MH] - =261.13.
步骤B：合成6-氨基-1H-吲哚-2-羧酸叔丁酯Step B: Synthesis of 6-amino-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000185
Figure PCTCN2020130361-appb-000185
室温下，将6-硝基-1H-吲哚-2-羧酸叔丁酯(200毫克，0.76毫摩尔)溶于乙酸乙酯中，加入钯碳(20毫克)，安装氢气球，室温反应9小时。At room temperature, dissolve 6-nitro-1H-indole-2-carboxylic acid tert-butyl ester (200 mg, 0.76 mmol) in ethyl acetate, add palladium on carbon (20 mg), install a hydrogen balloon, and react at room temperature 9 hours.
反应结束，过滤钯碳，滤饼用乙酸乙酯洗涤，滤液减压浓缩，干燥得46毫克6-氨基-1H-吲哚-2-羧酸叔丁酯，无需纯化，直接用于下一步反应。LCMS:RT＝3.16min,[M+H] +＝233.19。 After the reaction is complete, the palladium carbon is filtered, the filter cake is washed with ethyl acetate, the filtrate is concentrated under reduced pressure, and dried to obtain 46 mg of 6-amino-1H-indole-2-carboxylic acid tert-butyl ester, which is directly used in the next reaction without purification. . LCMS: RT = 3.16 min, [M+H] + =233.19.
步骤C：合成(S)-6-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step C: Synthesis of (S)-6-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000186
Figure PCTCN2020130361-appb-000186
室温下，将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酸(38毫克，0.09毫摩尔)、6-氨基-1H-吲哚-2-羧酸叔丁酯(20毫克，0.09毫摩尔)和2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(51毫克，0.13毫摩尔)加入N,N-二甲基甲酰胺(3.0毫升)中，滴加N,N-二异丙基乙胺(35毫克，0.27毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -Phenylpropionic acid (38 mg, 0.09 mmol), 6-amino-1H-indole-2-carboxylic acid tert-butyl ester (20 mg, 0.09 mmol) and 2-(7-benzotriazole oxide) )-N,N,N',N'-Tetramethylurea hexafluorophosphate (51 mg, 0.13 mmol) was added to N,N-dimethylformamide (3.0 mL), and N,N- Diisopropylethylamine (35 mg, 0.27 mmol) was dripped and reacted overnight at room temperature under N 2 protection.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到22毫克黄色固体(S)-6-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：37.3％)。LCMS:RT＝4.04min,[M+H] +＝655.39。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 22 mg of yellow solid (S)-6-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 37.3%). LCMS: RT = 4.04 min, [M+H] + =655.39.
步骤D：合成(S)-6-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Step D: Synthesis of (S)-6-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000187
Figure PCTCN2020130361-appb-000187
室温下，将(S)-6-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(22毫克，0.03毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-6-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-Phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (22 mg, 0.03 mmol) was added to dichloromethane (2.0 mL), and trifluoroacetic acid (0.5 mL) was added dropwise , React at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到9毫克淡黄色固体(S)-6-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：53.0％)。LCMS:RT＝3.42min,[M-H] -＝597.09。 1H NMR(500MHz,DMSO)δ12.87(s,1H),11.69(s,1H),10.28(s,1H),9.77(s,1H),7.98(s,1H),7.95(d,J＝2.3Hz,1H),7.82(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.57(d,J＝8.7Hz,1H),7.37–7.28(m,3H),7.25(t,J＝7.2Hz,1H),7.18(dd,J＝8.8,1.6Hz,1H),7.06–7.01(m,1H),5.42–5.35(m,1H),5.35–5.29(m,1H),3.32–3.27(m,2H),3.18–3.05(m,2H),2.08–1.92(m,2H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 9 mg of light yellow solid (S)-6-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3 -Dioxopiperazin-1-yl)-3-phenylpropionamido)-1H-indole-2-carboxylic acid (yield: 53.0%). LCMS: RT=3.42min, [MH] - =597.09. 1 H NMR (500MHz, DMSO) δ 12.87 (s, 1H), 11.69 (s, 1H), 10.28 (s, 1H), 9.77 (s, 1H), 7.98 (s, 1H), 7.95 (d, J =2.3Hz,1H),7.82(d,J=8.6Hz,1H),7.77(dd,J=8.6,2.3Hz,1H),7.57(d,J=8.7Hz,1H),7.37–7.28(m ,3H),7.25(t,J=7.2Hz,1H),7.18(dd,J=8.8,1.6Hz,1H),7.06–7.01(m,1H),5.42–5.35(m,1H),5.35– 5.29(m,1H), 3.32–3.27(m,2H), 3.18–3.05(m,2H), 2.08–1.92(m,2H).
实施例34Example 34
合成(S)-N-(4-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-((4-羟基-2-氧代-1,2-二氢喹啉-6-基)氨基)-3-氧代丙基)苯基)-4-氰基哌啶-1-甲酰胺Synthesis of (S)-N-(4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-((4-Hydroxy-2-oxo-1,2-dihydroquinolin-6-yl)amino)-3-oxopropyl)phenyl)-4-cyanopiperidine-1 -Formamide
Figure PCTCN2020130361-appb-000188
Figure PCTCN2020130361-appb-000188
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯Step A: Synthesis of (S)-3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxide Piperazin-1-yl) tert-butyl propionate
Figure PCTCN2020130361-appb-000189
Figure PCTCN2020130361-appb-000189
室温下，将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯(600毫克，1.1毫摩尔)和还原铁粉(618毫克，11.0毫摩尔)加入冰醋酸(12.0毫升)中，100摄氏度反应1小时。At room temperature, (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-Nitrophenyl) tert-butyl propionate (600 mg, 1.1 mmol) and reduced iron powder (618 mg, 11.0 mmol) were added to glacial acetic acid (12.0 mL) and reacted at 100 degrees Celsius for 1 hour.
反应结束，反应液用饱和碳酸钠调pH至8，垫硅藻土抽滤，滤饼用20毫升二氯甲烷洗涤，滤液用二氯甲烷(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/4)。得到400毫克黄色固体(S)-3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(收率：71.0％)。LCMS:RT＝3.10min,[M+H] +＝512.14。 At the end of the reaction, the pH of the reaction solution was adjusted to 8 with saturated sodium carbonate, filtered through a pad of diatomaceous earth, the filter cake was washed with 20 ml of dichloromethane, and the filtrate was extracted with dichloromethane (10 ml×3 times), and the organic phases were combined. The phase was washed with saturated brine (10 ml×2 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/4). Obtain 400 mg of yellow solid (S)-3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-di (Oxopiperazin-1-yl) tert-butyl propionate (yield: 71.0%). LCMS: RT = 3.10 min, [M+H] + = 512.14.
步骤B：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)基)氨基)苯基)丙酸叔丁酯Step B: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((phenoxycarbonyl)yl)amino)phenyl)tert-butyl propionate
Figure PCTCN2020130361-appb-000190
Figure PCTCN2020130361-appb-000190
N 2保护、冰浴下，向含有(S)-3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酸叔丁酯(400毫克，0.8毫摩尔)和三乙胺(326微升)的四氢呋喃(6.0毫升)中滴加氯甲酸苯酯(146微升)，滴毕，室温反应1小时。 Under the protection of N 2 and ice bath, it contains (S)-3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)- 2,3-dioxopiperazin-1-yl) tert-butyl propionate (400 mg, 0.8 mmol) and triethylamine (326 μl) in tetrahydrofuran (6.0 ml) were added dropwise to phenyl chloroformate ( 146 microliters), after dripping, react at room temperature for 1 hour.
反应结束，加水淬灭，混合液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/二氯甲烷＝1/1)。得到253毫克白色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)基)氨基)苯基)丙酸叔丁酯(收率：50.0％)。LCMS:RT＝4.04min,[M+H] +＝632.19。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (30 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/dichloromethane=1/1). Obtain 253 mg of white solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-butyl 3-(4-((phenoxycarbonyl)yl)amino)phenyl)propionate (yield: 50.0%). LCMS: RT = 4.04 min, [M+H] + =632.19.
步骤C：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰氨基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Cyanopiperidine-1-carboxamido)phenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000191
Figure PCTCN2020130361-appb-000191
室温下，向含有(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)基)氨基)苯基)丙酸叔丁酯(253毫克，0.4毫摩尔)和4-氰基哌啶(442毫克，4.0毫摩尔)的四氢呋喃(10.0毫升)中滴加N,N-二异丙基乙胺(518毫克，4.0毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Tert-butyl 3-(4-((phenoxycarbonyl)yl)amino)phenyl)propionate (253 mg, 0.4 mmol) and 4-cyanopiperidine (442 mg, 4.0 mmol) in tetrahydrofuran ( 10.0 ml) was added dropwise with N,N-diisopropylethylamine (518 mg, 4.0 mmol), after the dropping, the reaction was carried out at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到180毫克淡黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰氨基)苯基)丙酸叔丁酯(收率：69.4％)。LCMS:RT＝3.67min。After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 180 mg of light yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl) Tert-Butyl-3-(4-(4-cyanopiperidine-1-carboxamido)phenyl)propionate (yield: 69.4%). LCMS: RT = 3.67 min.
步骤D：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰氨基)苯基)丙酸Step D: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionic acid
Figure PCTCN2020130361-appb-000192
Figure PCTCN2020130361-appb-000192
室温下，将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰氨基)苯基)丙酸叔丁酯(180毫克，0.28毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Cyanopiperidine-1-carboxamido)phenyl) tert-butyl propionate (180 mg, 0.28 mmol) was added to dichloromethane (2.0 mL), and trifluoroacetic acid ( 0.5 ml), react at room temperature for 3 hours.
反应结束，减压浓缩得到160毫克黄色油状物(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰氨基)苯基)丙酸，无需纯化，直接用于下一步反应。LCMS:RT＝3.24min,[M-H] -＝590.11。 After the reaction was over, concentrated under reduced pressure to obtain 160 mg of yellow oil (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper Azin-1-yl)-3-(4-(4-cyanopiperidine-1-carboxamido)phenyl)propionic acid was used directly in the next reaction without purification. LCMS: RT = 3.24 min, [MH] - = 590.11.
步骤E：合成(S)-N-(4-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-((4-羟基-2-氧代-1,2-二氢喹啉-6-基)氨基)-3-氧代丙基)苯基)-4-氰基哌啶-1-甲酰胺Step E: Synthesis of (S)-N-(4-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine- 1-yl)-3-((4-hydroxy-2-oxo-1,2-dihydroquinolin-6-yl)amino)-3-oxopropyl)phenyl)-4-cyanopiper Pyridine-1-carboxamide
Figure PCTCN2020130361-appb-000193
Figure PCTCN2020130361-appb-000193
室温下，将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰氨基)苯基)丙酸(160毫克，0.27毫摩尔)、6-氨基-4-羟基-2(1H)-喹啉酮(53毫克，0.30毫摩尔)和2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(206毫克，0.54毫摩尔)加入N,N-二甲基甲酰胺(4.0毫升)中，滴加N,N-二异丙基乙胺(105毫克，0.81毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionic acid (160 mg, 0.27 mmol), 6-amino-4-hydroxy-2(1H)-quinolinone (53 Mg, 0.30 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (206 mg, 0.54 mmol) add N,N -In dimethylformamide (4.0 ml), add N,N-diisopropylethylamine (105 mg, 0.81 mmol) dropwise, and after the dropwise, react under the protection of N 2 at room temperature overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：纯乙酸乙酯。得到54毫克白固体(S)-N-(4-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-((4-羟基-2-氧代-1,2-二氢喹啉-6-基)氨基)-3-氧代丙基)苯基)-4-氰基哌啶-1-甲酰胺(收率：26.7％)。LCMS:RT＝3.16min,[M+H] +＝750.20。 1H NMR(400MHz,DMSO)δ11.36(s,1H),11.21(s,1H),10.30(s,1H),9.79(s,1H),8.53(s,1H),8.16(d,J＝1.9Hz,1H),7.95(d,J＝2.3Hz,1H),7.82(d,J＝8.7Hz,1H),7.78–7.74(m,1H),7.62(dd,J＝8.9,2.3Hz,1H),7.40(d,J＝8.5Hz,1H),7.21(d,J＝8.9Hz,1H),7.14(d,J＝9.2Hz,1H),6.66(s,1H),5.74(d,J＝1.0Hz,1H),5.32(t,J＝4.8Hz,1H),5.26(dd,J＝10.2,5.6Hz,1H),3.32–3.15(m,4H),3.14–2.92(m,4H),2.08–1.95(m,3H),1.95–1.79(m,2H),1.76–1.60(m,2H)。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: pure ethyl acetate. 54 mg of white solid (S)-N-(4-(2-(4-(5-chloro-2-(1H-tetra (Azol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-((4-hydroxy-2-oxo-1,2-dihydroquinolin-6-yl )Amino)-3-oxopropyl)phenyl)-4-cyanopiperidine-1-carboxamide (yield: 26.7%). LCMS: RT = 3.16 min, [M+H] + =750.20. 1 H NMR (400MHz, DMSO) δ 11.36 (s, 1H), 11.21 (s, 1H), 10.30 (s, 1H), 9.79 (s, 1H), 8.53 (s, 1H), 8.16 (d, J ＝1.9Hz,1H),7.95(d,J＝2.3Hz,1H),7.82(d,J＝8.7Hz,1H),7.78–7.74(m,1H),7.62(dd,J＝8.9,2.3Hz ,1H),7.40(d,J=8.5Hz,1H),7.21(d,J=8.9Hz,1H),7.14(d,J=9.2Hz,1H),6.66(s,1H),5.74(d ,J=1.0Hz,1H),5.32(t,J=4.8Hz,1H), 5.26(dd,J=10.2,5.6Hz,1H), 3.32–3.15(m,4H),3.14–2.92(m, 4H), 2.08–1.95(m,3H), 1.95–1.79(m,2H), 1.76–1.60(m,2H).
实施例35Example 35
合成5-((2S)-3-(4-(3-((1,4-二恶烷-2-基)甲基)脲基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((2S)-3-(4-(3-((1,4-dioxan-2-yl)methyl)ureido)phenyl)-2-(4-(5-chloro- 2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000194
Figure PCTCN2020130361-appb-000194
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-2-氨基-3-(4-硝基苯基)丙酸叔丁酯Step A: Synthesis of tert-butyl (S)-2-amino-3-(4-nitrophenyl)propionate
Figure PCTCN2020130361-appb-000195
Figure PCTCN2020130361-appb-000195
N 2保护、冰浴下，向含有(S)-2-氨基-3-(4-硝基苯基)丙酸(20.0克，95.2毫摩尔)的醋酸叔丁酯(200毫升)中滴加70％的高氯酸(8.2毫升)，滴毕，室温反应过夜。 Under N 2 protection and ice bath, add dropwise to tert-butyl acetate (200 ml) containing (S)-2-amino-3-(4-nitrophenyl)propionic acid (20.0 g, 95.2 mmol) 70% perchloric acid (8.2 ml) was dripped and reacted overnight at room temperature.
反应结束，加乙酸乙酯稀释，用饱和碳酸钠调pH至9，混合液用乙酸乙酯(150毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(100毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝5/1)。得到15.2克黄色油状物(S)-2-氨基-3-(4-硝基苯基)丙酸丁酯(收率：60.0％)。LCMS:RT＝2.05min。After the reaction is over, add ethyl acetate to dilute, adjust the pH to 9 with saturated sodium carbonate, extract the mixture with ethyl acetate (150 ml×3 times), combine the organic phases, and use saturated brine (100 ml×2 times). ) Wash, then dry with anhydrous sodium sulfate, and finally concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=5/1). 15.2 g of butyl (S)-2-amino-3-(4-nitrophenyl)propionate was obtained as a yellow oil (yield: 60.0%). LCMS: RT = 2.05 min.
步骤B：合成(S)-2-(2-甲氧基-2-氧代乙酰氨基)-3-(4-硝基苯基)丙酸叔丁酯Step B: Synthesis of tert-butyl (S)-2-(2-methoxy-2-oxoacetamido)-3-(4-nitrophenyl)propionate
Figure PCTCN2020130361-appb-000196
Figure PCTCN2020130361-appb-000196
N 2保护、冰浴下，向含有(S)-2-氨基-3-(4-硝基苯基)丙酸丁酯(15.2克，57.1毫摩尔)和三乙胺(6.9克，68.5毫摩尔)的二氯甲烷(150毫升)中滴加草酰氯单甲酯(7.7克，62.8毫摩尔)，滴毕，室温反应30分钟。 Under the protection of N 2 and ice bath, add (S)-2-amino-3-(4-nitrophenyl) butyl propionate (15.2 g, 57.1 mmol) and triethylamine (6.9 g, 68.5 mmol). Methyl oxalyl chloride (7.7 g, 62.8 mmol) was added dropwise to dichloromethane (150 ml) in mol). After the dripping was completed, the reaction was carried out at room temperature for 30 minutes.
反应结束，加水淬灭，混合液用二氯甲烷(100毫升×3次)萃取。合并有机相，有机相先用饱和食盐水(50毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩得到13.3克淡黄色油状物(S)-2-(2-甲氧基-2-氧代乙酰氨基)-3-(4-硝基苯基)丙酸叔丁酯，无需纯化，直接用于下一步反应。LCMS:RT＝3.91min,[M-H] -＝351.12。 After the reaction was completed, it was quenched by adding water, and the mixture was extracted with dichloromethane (100 ml×3 times). The organic phases were combined, and the organic phase was washed with saturated brine (50 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure to obtain 13.3 g of light yellow oil (S)-2-(2-methoxy Tert-butyl 2-oxoacetamido)-3-(4-nitrophenyl)propionate, without purification, was directly used in the next reaction. LCMS: RT = 3.91 min, [MH] - = 351.12.
步骤C：合成(S)-2-((1-(叔丁氧基)-3-(4-硝基苯基)-1-氧代丙烷-2-基)氨基)-2-氧代乙酸Step C: Synthesis of (S)-2-((1-(tert-butoxy)-3-(4-nitrophenyl)-1-oxopropan-2-yl)amino)-2-oxoacetic acid
Figure PCTCN2020130361-appb-000197
Figure PCTCN2020130361-appb-000197
冰浴下，向含有(S)-2-(2-甲氧基-2-氧代乙酰氨基)-3-(4-硝基苯基)丙酸叔丁酯(13.3克，37.7毫摩尔)的四氢呋喃(160毫升)中滴加氢氧化锂(3.2克，75.4毫摩尔)水溶液(80毫升)，滴毕即反应结束。Under an ice bath, add tert-butyl (S)-2-(2-methoxy-2-oxoacetamido)-3-(4-nitrophenyl)propionate (13.3 g, 37.7 mmol) Lithium hydroxide (3.2 g, 75.4 mmol) aqueous solution (80 ml) was added dropwise to the tetrahydrofuran (160 ml), and the reaction was completed when the dropping was completed.
反应结束，加水淬灭，用稀盐酸水溶液(0.5摩尔/升)调pH至4，混合液用乙酸乙酯(80毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(50毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。最后减压浓缩得到10.2克淡黄色油状物(S)-2-((1-(叔丁氧基)-3-(4-硝基苯基)-1-氧代丙烷-2-基)氨基)-2-氧代乙酸，无需纯化，直接用于下一步反应。LCMS:RT＝4.32min,[M-H] -＝337.16。 After the reaction is over, add water to quench, adjust the pH to 4 with dilute aqueous hydrochloric acid (0.5 mol/L), extract the mixture with ethyl acetate (80 ml×3 times), combine the organic phases, and use saturated brine (50 (Ml×2 times) washing, then drying with anhydrous sodium sulfate, and finally concentrating under reduced pressure. Finally, it was concentrated under reduced pressure to obtain 10.2 g of light yellow oil (S)-2-((1-(tert-butoxy)-3-(4-nitrophenyl)-1-oxopropan-2-yl)amino )-2-oxoacetic acid, without purification, directly used in the next reaction LCMS: RT = 4.32 min, [MH] - = 337.16.
步骤D：合成N，N-二烯丙基-4-氯-2-硝基苯胺Step D: Synthesis of N,N-diallyl-4-chloro-2-nitroaniline
Figure PCTCN2020130361-appb-000198
Figure PCTCN2020130361-appb-000198
室温下，将5-氯-2-氟硝基苯(20.0克，113.9毫摩尔)、二烯丙基胺(16.6克，170.9毫摩尔)和碳酸钾(32.0克，231.9毫摩尔)加入N,N-二甲基甲酰胺(200毫升)中，N 2保护下，80摄氏度反应3小时。 At room temperature, add 5-chloro-2-fluoronitrobenzene (20.0 g, 113.9 mmol), diallylamine (16.6 g, 170.9 mmol) and potassium carbonate (32.0 g, 231.9 mmol) to N, In N-dimethylformamide (200 ml), under the protection of N 2 , the reaction was carried out at 80 degrees Celsius for 3 hours.
反应结束，垫无水硫酸钠抽滤，滤饼用200毫升乙酸乙酯洗涤，滤液用乙酸乙酯(100毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(100毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩得到27.5克棕色油状物N，N-二烯丙基-4-氯-2-硝基苯胺，无需纯化，直接用于下一步反应。LCMS:RT＝4.58min,[M+H] +＝253.07。 After the reaction was over, the pad was suction filtered with anhydrous sodium sulfate, the filter cake was washed with 200 ml of ethyl acetate, the filtrate was extracted with ethyl acetate (100 ml × 3 times), the organic phases were combined, and the organic phase was first used with saturated brine (100 ml × 2 times) washing, then drying with anhydrous sodium sulfate, and finally concentrating under reduced pressure to obtain 27.5 g of brown oil N,N-diallyl-4-chloro-2-nitroaniline, which is used directly in the next step without purification. reaction. LCMS: RT = 4.58 min, [M+H] + =253.07.
步骤E：合成N 1,N 1-二烯丙基-4-氯苯-1,2-二胺 Step E: Synthesis of N 1 ,N 1 -diallyl-4-chlorobenzene-1,2-diamine
Figure PCTCN2020130361-appb-000199
Figure PCTCN2020130361-appb-000199
室温下，将N,N-二烯丙基-4-氯-2-硝基苯胺(27.5克，108.8毫摩尔)加入乙酸乙酯中，冰浴下，分批加入氯化亚锡二水合物(122.8克，544.0毫摩尔)，N 2保护下，室温反应过夜。 At room temperature, add N,N-diallyl-4-chloro-2-nitroaniline (27.5 g, 108.8 mmol) to ethyl acetate, and add stannous chloride dihydrate in batches under ice bath (122.8 g, 544.0 mmol), under N 2 protection, react at room temperature overnight.
反应结束，用饱和碳酸氢钠水溶液淬灭，再加入过量碳酸氢钠固体，调pH至弱碱性，垫硅藻土抽滤，滤饼用300毫升乙酸乙酯洗涤，滤液用乙酸乙酯(100毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(200毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：纯正己烷)。得到11.4克棕色油状物N 1,N 1-二烯丙基-4-氯苯-1,2-二胺(收率：47.1％)。LCMS:RT＝4.44min,[M+H] +＝223.22。 After the reaction is over, it is quenched with saturated sodium bicarbonate aqueous solution, and then excess sodium bicarbonate solid is added to adjust the pH to weakly alkaline. The filter cake is suction filtered with a pad of celite. The filter cake is washed with 300 ml of ethyl acetate, and the filtrate is washed with ethyl acetate ( 100 ml × 3 times), and the organic phases were combined. The organic phase was washed with saturated brine (200 ml × 2 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: pure n-hexane). 11.4 g of brown oily N 1 ,N 1 -diallyl-4-chlorobenzene-1,2-diamine was obtained (yield: 47.1%). LCMS: RT = 4.44 min, [M+H] + =223.22.
步骤F：合成(S)-2-(2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-硝基苯基)丙酸叔丁酯Step F: Synthesis of (S)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido)-3-(4-nitrobenzene Base) tert-butyl propionate
Figure PCTCN2020130361-appb-000200
Figure PCTCN2020130361-appb-000200
室温下，将油状物(S)-2-((1-(叔丁氧基)-3-(4-硝基苯基)-1-氧代丙烷-2-基)氨基)-2-氧代乙酸(15.7克，46.4毫摩尔)、N 1,N 1-二烯丙基-4-氯苯-1,2-二胺(11.4克，51.2毫摩尔)和2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(35.3克，92.8毫摩尔)加入N,N-二甲基甲酰胺(300毫升)中，冰浴下滴加N,N-二异丙基乙胺(18.0克，138.3毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, the oily (S)-2-((1-(tert-butoxy)-3-(4-nitrophenyl)-1-oxopropan-2-yl)amino)-2-oxy Acetic acid (15.7 g, 46.4 mmol), N 1 , N 1 -diallyl-4-chlorobenzene-1,2-diamine (11.4 g, 51.2 mmol) and 2-(7-oxybenzo Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (35.3g, 92.8mmol) was added to N,N-dimethylformamide (300ml), under ice bath N,N-diisopropylethylamine (18.0 g, 138.3 mmol) was added dropwise, and after the dropping was completed, the reaction was carried out at room temperature overnight under the protection of N 2.
反应结束，加水淬灭，混合液用乙酸乙酯(200毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(100毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝5/1)。得到20.0克黄色油状物(S)-2-(2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-硝基苯基)丙酸叔丁酯(收率：79.4％)。LCMS:RT＝5.21min,[M-H] -＝541.14。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (200 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (100 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=5/1). Obtain 20.0 grams of yellow oil (S)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido)-3-(4-nitro Tert-butyl phenyl)propionate (yield: 79.4%). LCMS: RT = 5.21 min, [MH] - = 541.14.
步骤G：合成(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯Step G: Synthesis of (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4 -Nitrophenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000201
Figure PCTCN2020130361-appb-000201
室温下，将(S)-2-(2-((5-氯-2-(二烯丙基氨基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-硝基苯基)丙酸叔丁酯(20.0克，36.8毫摩尔)和碳酸钾(31.0克，224.3毫摩尔)加入三颈瓶中，N 2保护下，用双排针加入乙腈(250毫升)，再用注射器注入1,2-二溴乙烷(42.1克，224.3毫摩尔)，100摄氏度反应过夜。 At room temperature, (S)-2-(2-((5-chloro-2-(diallylamino)phenyl)amino)-2-oxoacetamido)-3-(4-nitrobenzene Base) tert-butyl propionate (20.0 g, 36.8 mmol) and potassium carbonate (31.0 g, 224.3 mmol) were added to a three-necked flask, under N 2 protection, add acetonitrile (250 ml) with a double-row needle, and then use The syringe was injected with 1,2-dibromoethane (42.1 g, 224.3 mmol) and reacted at 100 degrees Celsius overnight.
反应结束，垫无水硫酸钠抽滤，滤饼用200毫升乙酸乙酯洗涤，滤液用乙酸乙酯(100毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(100毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝2/1)。得到10.5克黄色固体(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯(收率：49.8％)。LCMS:RT＝4.44min,[M+H] +＝569.23。 After the reaction was over, the pad was suction filtered with anhydrous sodium sulfate, the filter cake was washed with 200 ml of ethyl acetate, the filtrate was extracted with ethyl acetate (100 ml × 3 times), the organic phases were combined, and the organic phase was first used with saturated brine (100 ml × 2 times) washing, then drying with anhydrous sodium sulfate, and finally concentrating under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=2/1). 10.5 g of yellow solid (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-( Tert-Butyl 4-nitrophenyl)propionate (yield: 49.8%). LCMS: RT = 4.44 min, [M+H] + =569.23.
步骤H：合成(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯Step H: Synthesis of (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-nitrophenyl) Tert-butyl propionate
Figure PCTCN2020130361-appb-000202
Figure PCTCN2020130361-appb-000202
室温下，向含有(S)-2-(4-(5-氯-2-(二烯丙基氨基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯(10.50克，18.5毫摩尔)的二氯甲烷(150毫升)中加入1,3-二甲基巴比妥酸(17.30克，111.0毫摩尔)和四三苯基膦钯(0.85克，0.7毫摩尔)，40摄氏度反应过夜。At room temperature, it contains (S)-2-(4-(5-chloro-2-(diallylamino)phenyl)-2,3-dioxopiperazin-1-yl)-3-( 4-nitrophenyl) tert-butyl propionate (10.50 g, 18.5 mmol) in dichloromethane (150 ml) was added 1,3-dimethylbarbituric acid (17.30 g, 111.0 mmol) and Tetratriphenylphosphine palladium (0.85 g, 0.7 mmol) was reacted overnight at 40 degrees Celsius.
反应结束，加二氯甲烷稀释，用饱和碳酸钠调pH至8，混合液用二氯甲烷(100毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(50毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到8.4克黄色固体(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯(收率：92.9％)。LCMS:RT＝3.98min，[M+H] +＝489.15。 After the reaction is over, dilute with dichloromethane, adjust the pH to 8 with saturated sodium carbonate, extract the mixture with dichloromethane (100ml×3 times), combine the organic phases, and use saturated brine (50ml×2 times). ) Wash, then dry with anhydrous sodium sulfate, and finally concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 8.4 grams of yellow solid (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-nitrophenyl) ) Tert-butyl propionate (yield: 92.9%). LCMS: RT = 3.98 min, [M+H] + =489.15.
步骤I：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯Step I: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-Nitrophenyl) tert-butyl propionate
Figure PCTCN2020130361-appb-000203
Figure PCTCN2020130361-appb-000203
室温下，向含有(S)-2-(4-(2-氨基-5-氯苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯(8.4克，17.2毫摩尔)的冰醋酸(80毫升)中加入原甲酸三乙酯(12.7克，85.9毫摩尔)和叠氮化钠(3.9克，60.0毫摩尔)，80摄氏度反应5小时。At room temperature, add (S)-2-(4-(2-amino-5-chlorophenyl)-2,3-dioxopiperazin-1-yl)-3-(4-nitrophenyl) ) Tert-butyl propionate (8.4 g, 17.2 mmol) in glacial acetic acid (80 mL) was added triethyl orthoformate (12.7 g, 85.9 mmol) and sodium azide (3.9 g, 60.0 mmol), React at 80 degrees Celsius for 5 hours.
反应结束，加入20毫升水，逐滴加入亚硝酸钠(250毫克/毫升)至无气泡产生，加80毫升乙酸乙酯分散，抽滤，滤饼用30毫升乙酸乙酯洗涤，干燥得2.5克类白色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯。(收率：26.8％)。LCMS:RT＝4.08min。After the reaction is over, add 20 ml of water, add sodium nitrite (250 mg/ml) dropwise until no bubbles are generated, add 80 ml of ethyl acetate to disperse, filter with suction, wash the filter cake with 30 ml of ethyl acetate, and dry to obtain 2.5 g Off-white solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-Nitrophenyl) tert-butyl propionate. (Yield: 26.8%). LCMS: RT = 4.08 min.
步骤J：合成(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸Step J: Synthesis of (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-Nitrophenyl)propionic acid
Figure PCTCN2020130361-appb-000204
Figure PCTCN2020130361-appb-000204
室温下，向含有(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸叔丁酯(2.5克，4.6毫摩尔)的二氯甲烷(24.0毫升)中滴加三氟乙酸(6.0毫升)，室温反应5小时。At room temperature, it contains (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)- Trifluoroacetic acid (6.0 mL) was added dropwise to tert-butyl 3-(4-nitrophenyl) propionate (2.5 g, 4.6 mmol) in dichloromethane (24.0 mL), and reacted at room temperature for 5 hours.
反应结束，减压浓缩得到1.8克黄色固体(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸，无需纯化，直接用于下一步反应。LCMS:RT＝3.48min,[M-H] -＝484.07。 After the reaction is over, it is concentrated under reduced pressure to obtain 1.8 g of yellow solid (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine -1-yl)-3-(4-nitrophenyl)propionic acid, without purification, was directly used in the next reaction. LCMS: RT = 3.48 min, [MH] - =484.07.
步骤K：合成5-硝基-1H-吲哚-1,2-二羧酸二叔丁酯Step K: Synthesis of 5-nitro-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000205
Figure PCTCN2020130361-appb-000205
室温下，将5-硝基吲哚-2-甲酸(8.0克，38.8毫摩尔)和二碳酸二叔丁酯(38.1克，174.6毫摩尔)加入N,N-二甲基甲酰胺(40毫升)中，冰浴下，分批加入4-二甲氨基吡啶(3.8克，31.1毫摩尔)，N 2保护下，室温反应过夜。 At room temperature, add 5-nitroindole-2-carboxylic acid (8.0 g, 38.8 mmol) and di-tert-butyl dicarbonate (38.1 g, 174.6 mmol) to N,N-dimethylformamide (40 mL ), under ice bath, add 4-dimethylaminopyridine (3.8 g, 31.1 mmol) in batches, and react at room temperature overnight under N 2 protection.
反应结束，加水淬灭，加入100毫升乙酸乙酯，垫硅藻土抽滤，滤饼用50毫升乙酸乙酯洗涤，滤液用乙酸乙酯(100毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(100毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩得到11.0克淡黄色油状物5-硝基-1H-吲哚-1,2-二羧酸二叔丁酯，无需纯化，直接用于下一步反应。LCMS:RT＝5.01min。After the reaction is over, add water to quench, add 100 ml of ethyl acetate, filter with Celite pad, wash the filter cake with 50 ml of ethyl acetate, and extract the filtrate with ethyl acetate (100 ml×3 times). Combine the organic phases. The phase was first washed with saturated brine (100 ml×2 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure to obtain 11.0 g of light yellow oily 5-nitro-1H-indole-1,2-dicarboxylate The di-tert-butyl ester is directly used in the next reaction without purification. LCMS: RT = 5.01 min.
步骤L：合成5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯Step L: Synthesis of 5-amino-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000206
Figure PCTCN2020130361-appb-000206
室温下，将5-硝基-1H-吲哚-1,2-二羧酸二叔丁酯(11.0克，30.4毫摩尔l)溶于乙醇中，加入钯碳(1.1克)，安装氢气球，室温反应9小时。At room temperature, dissolve 5-nitro-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (11.0 g, 30.4 mmol l) in ethanol, add palladium on carbon (1.1 g), and install a hydrogen balloon , React at room temperature for 9 hours.
反应结束，过滤钯碳，滤饼用乙醇洗涤，滤液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到8.2克黄色固体5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯(收率：81.0％)。LCMS:RT＝4.09min,,[M+H] +＝333.20。 After the reaction was completed, the palladium carbon was filtered, the filter cake was washed with ethanol, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). 8.2 g of yellow solid 5-amino-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 81.0%) was obtained. LCMS: RT = 4.09 min,, [M+H] + =333.20.
步骤M：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-硝基苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step M: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-nitrophenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000207
Figure PCTCN2020130361-appb-000207
室温下，将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸(1.8克，3.7毫摩尔)、5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯(1.4克，4.2毫摩尔)和2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(2.8克，7.4毫摩尔)加入N,N-二甲基甲酰胺(20.0毫升)中，滴加N,N-二异丙基乙胺(1.4克，11.2毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-nitrophenyl) propionic acid (1.8 g, 3.7 mmol), 5-amino-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (1.4 g, 4.2 mmol) and 2 -(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.8 g, 7.4 mmol) is added to N,N-dimethylformamide (20.0 (Ml), N,N-diisopropylethylamine (1.4 g, 11.2 mmol) was added dropwise, after the dropping, the reaction was carried out at room temperature overnight under the protection of N 2.
反应结束，加水淬灭，混合液用乙酸乙酯(50毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(30毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到1.3克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-硝基苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：43.2％)。LCMS:RT＝4.45min,[M-H]-＝798.13。After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (50 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (30 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 1.3 g of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- (Yl)-3-(-4-nitrophenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 43.2%). LCMS: RT=4.45 min, [M-H]-=798.13.
步骤N：合成(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step N: Synthesis of (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3 -Dioxopiperazin-1-yl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000208
Figure PCTCN2020130361-appb-000208
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-硝基苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(1.3克，1.6毫摩尔)和还原铁粉(0.9克，16.1毫摩尔)加入甲醇(15.0毫升)中，滴加冰醋酸(1.0毫升)，65摄氏度反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-nitrophenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (1.3 g, 1.6 mmol) and reduced iron powder (0.9 g, 16.1 mmol) was added to methanol (15.0 ml), glacial acetic acid (1.0 ml) was added dropwise, and the reaction was carried out at 65 degrees Celsius for 3 hours.
反应结束，向反应液中加入10毫升乙酸乙酯稀释，垫硅藻土抽滤，滤饼用乙酸乙酯洗涤，滤液用饱和碳酸氢钠调pH至10，混合液用乙酸乙酯(50毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到622毫克黄色固体(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：50.0％)。LCMS:RT＝4.05min,[M-H] -＝768.20。 At the end of the reaction, add 10 ml of ethyl acetate to the reaction solution for dilution, pad diatomaceous earth for suction filtration, wash the filter cake with ethyl acetate, adjust the pH of the filtrate to 10 with saturated sodium bicarbonate, and use ethyl acetate (50 ml × 3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml × 2 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 622 mg of yellow solid (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2, 3-Dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 50.0%). LCMS: RT = 4.05 min, [MH] - = 768.20.
步骤O：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step O: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000209
Figure PCTCN2020130361-appb-000209
N 2保护、冰浴下，向含有(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(622毫克，0.8毫摩尔)和三乙胺(336微升)的四氢呋喃(6.0毫升)中滴加氯甲酸苯酯(202微升)，滴毕，室温反应1小时。 Under the protection of N 2 and ice bath, it will contain (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)benzene (Yl)-2,3-dioxopiperazin-1-yl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (622 mg, 0.8 mmol) and triethylamine Phenyl chloroformate (202 microliters) was added dropwise to (336 microliters) of tetrahydrofuran (6.0 mL). After the dripping, the reaction was carried out at room temperature for 1 hour.
反应结束，加水淬灭，混合液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/二氯甲烷＝1/1)。得到480毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：67.4％)。LCMS:RT＝4.52min。After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (30 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/dichloromethane=1/1). Obtain 480 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 67.4%). LCMS: RT = 4.52 min.
步骤P：合成5-((2S)-3-(4-(3-((1,4-二恶烷-2-基)甲基)脲基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step P: Synthesis of 5-((2S)-3-(4-(3-((1,4-dioxan-2-yl)methyl)ureido)phenyl)-2-(4-(5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester
Figure PCTCN2020130361-appb-000210
Figure PCTCN2020130361-appb-000210
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和1,4-二恶烷-2-甲胺盐酸盐(207毫克，1.4毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) And 1,4-dioxane-2-methylamine hydrochloride (207 mg, 1.4 mmol) in tetrahydrofuran (3.0 ml) was added dropwise N,N-diisopropylethylamine (174 mg, 1.4 mmol) ), after dripping, react overnight at 65 degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/4)。得到50毫克白色固体5-((2S)-3-(4-(3-((1,4-二恶烷-2-基)甲基)脲基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：39.1％)。LCMS:RT＝4.19min,[M+H] +＝913.33。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/4). Obtain 50 mg of white solid 5-((2S)-3-(4-(3-((1,4-dioxan-2-yl)methyl)ureido)phenyl)-2-(4-( 5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxy Di-tert-butyl acid (yield: 39.1%). LCMS: RT = 4.19 min, [M+H] + =913.33.
步骤Q：合成5-((2S)-3-(4-(3-((1,4-二恶烷-2-基)甲基)脲基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Step Q: Synthesis of 5-((2S)-3-(4-(3-((1,4-dioxan-2-yl)methyl)ureido)phenyl)-2-(4-(5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000211
Figure PCTCN2020130361-appb-000211
室温下，将5-((2S)-3-(4-(3-((1,4-二恶烷-2-基)甲基)脲基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(50毫克，0.05毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, add 5-((2S)-3-(4-(3-((1,4-dioxan-2-yl)methyl)ureido)phenyl)-2-(4-(5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester (50 mg, 0.05 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到24毫克白色固体5-((2S)-3-(4-(3-((1,4-二恶烷-2-基)甲基)脲基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸(收率：63.4％)。LCMS:RT＝3.20min,[M-H] -＝755.05。 1H NMR(500MHz,DMSO)δ12.95(s,1H),11.75(s,1H),10.16(s,1H),9.81(s,1H),8.53(s,1H),8.02(s,1H),7.97(d,J＝2.2Hz,1H),7.83(d,J＝8.6Hz,1H),7.78(dd,J＝8.6,2.3Hz,1H),7.39(d,J＝8.9Hz,1H),7.34(dd,J＝9.7,2.8Hz,3H),7.15(d,J＝5.3Hz,2H),7.08(d,J＝1.6Hz,1H),6.23(t,J＝5.4Hz,1H),5.29(dd,J＝10.0,5.3Hz,1H),3.81–3.69(m,3H),3.67–3.58(m,2H),3.58–3.49(m,2H),3.49–3.42(m,2H),3.28–3.19(m,2H),3.09–2.97(m,2H),2.12–1.91(m,2H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 24 mg of white solid 5-((2S)-3-(4-(3-((1,4-dioxan-2-yl)methyl)ureido)phenyl )-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indyl Dole-2-carboxylic acid (yield: 63.4%). LCMS: RT = 3.20 min, [MH] - =755.05. 1 H NMR (500MHz, DMSO) δ 12.95 (s, 1H), 11.75 (s, 1H), 10.16 (s, 1H), 9.81 (s, 1H), 8.53 (s, 1H), 8.02 (s, 1H) ), 7.97 (d, J = 2.2 Hz, 1H), 7.83 (d, J = 8.6 Hz, 1H), 7.78 (dd, J = 8.6, 2.3 Hz, 1H), 7.39 (d, J = 8.9 Hz, 1H) ), 7.34 (dd, J = 9.7, 2.8 Hz, 3H), 7.15 (d, J = 5.3 Hz, 2H), 7.08 (d, J = 1.6 Hz, 1H), 6.23 (t, J = 5.4 Hz, 1H ), 5.29(dd,J=10.0,5.3Hz,1H), 3.81–3.69(m,3H), 3.67–3.58(m,2H), 3.58–3.49(m,2H), 3.49–3.42(m,2H) ), 3.28–3.19(m,2H), 3.09–2.97(m,2H), 2.12–1.91(m,2H).
实施例36Example 36
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((R)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000212
Figure PCTCN2020130361-appb-000212
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000213
Figure PCTCN2020130361-appb-000213
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和(R)-3-氨基丁醇(121毫克，1.4毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) And (R)-3-aminobutanol (121 mg, 1.4 mmol) in tetrahydrofuran (3.0 mL) was added dropwise N,N-diisopropylethylamine (174 mg, 1.4 mmol), dripping finished, 65 React overnight in degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到60毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：48.5％)。LCMS:RT＝4.15min,[M-H] -＝883.52。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 60 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((R)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert Butyl ester (yield: 48.5%). LCMS: RT = 4.15 min, [MH] - = 883.52.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000214
Figure PCTCN2020130361-appb-000214
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(60毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl The ester (60 mg, 0.07 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到33毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：64.7％)。LCMS:RT＝3.19min,[M-H] -＝727.19。 1H NMR(500MHz,DMSO)δ12.92(s,1H),11.72(s,1H),10.13(s,1H),9.80(s,1H),8.31(s,1H),8.02(s,1H),7.97(d,J＝2.2Hz,1H),7.83(d,J＝8.6Hz,1H),7.78(dd,J＝8.6,2.3Hz,1H),7.37(dd,J＝24.4,8.6Hz,4H),7.14(d,J＝8.4Hz,2H),7.08(d,J＝1.7Hz,1H),6.01(d,J＝7.7Hz,1H),5.29(dd,J＝10.0,5.7Hz,1H),3.78(dd,J＝16.1,8.9Hz,3H),3.45(d,J＝10.3Hz,4H),3.22(dd,J＝13.8,6.2Hz,2H),3.01(dd,J＝14.9,10.7Hz,2H),1.10(d,J＝6.6Hz,3H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 33 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-((R)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2 -Carboxylic acid (yield: 64.7%). LCMS: RT = 3.19 min, [MH] - =727.19. 1 H NMR (500MHz, DMSO) δ 12.92 (s, 1H), 11.72 (s, 1H), 10.13 (s, 1H), 9.80 (s, 1H), 8.31 (s, 1H), 8.02 (s, 1H) ), 7.97 (d, J = 2.2 Hz, 1H), 7.83 (d, J = 8.6 Hz, 1H), 7.78 (dd, J = 8.6, 2.3 Hz, 1H), 7.37 (dd, J = 24.4, 8.6 Hz , 4H), 7.14 (d, J = 8.4 Hz, 2H), 7.08 (d, J = 1.7 Hz, 1H), 6.01 (d, J = 7.7 Hz, 1H), 5.29 (dd, J = 10.0, 5.7 Hz ,1H), 3.78 (dd, J = 16.1, 8.9 Hz, 3H), 3.45 (d, J = 10.3 Hz, 4H), 3.22 (dd, J = 13.8, 6.2 Hz, 2H), 3.01 (dd, J = 14.9, 10.7 Hz, 2H), 1.10 (d, J = 6.6 Hz, 3H).
实施例37Example 37
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((S)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000215
Figure PCTCN2020130361-appb-000215
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000216
Figure PCTCN2020130361-appb-000216
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和(S)-3-氨基丁醇(121毫克，1.4毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) And (S)-3-aminobutanol (121 mg, 1.4 mmol) in tetrahydrofuran (3.0 ml) was added dropwise N,N-diisopropylethylamine (174 mg, 1.4 mmol), dripping finished, 65 React overnight in degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到55毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-4-羟基丁-2-基)脲基)苯基) 丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：44.5％)。LCMS:RT＝4.15min,[M-H] -＝883.29。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 55 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((S)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert Butyl ester (yield: 44.5%). LCMS: RT = 4.15 min, [MH] - = 883.29.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000217
Figure PCTCN2020130361-appb-000217
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(55毫克，0.06毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl The ester (55 mg, 0.06 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到25毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-4-羟基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：49.1％)。LCMS:RT＝3.20min,[M-H] -＝727.15。 1H NMR(400MHz,DMSO)δ12.91(s,1H),11.73(s,1H),10.13(s,1H),9.79(s,1H),8.30(s,1H),8.01(d,J＝1.4Hz,1H),7.96(d,J＝2.3Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝9.0,1.8Hz,1H),7.42–7.31(m,3H),7.13(d,J＝7.7Hz,2H),7.07(d,J＝2.2Hz,1H),6.00(d,J＝8.0Hz,1H),5.29(dd,J＝9.8,5.6Hz,1H),3.77(dd,J＝20.3,8.9Hz,3H),3.53–3.40(m,4H),3.20(d,J＝10.0Hz,2H),2.98(s,2H),1.09(d,J＝6.6Hz,3H). When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 25 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-((S)-4-hydroxybut-2-yl)ureido)phenyl)propionamido)-1H-indole-2 -Carboxylic acid (yield: 49.1%). LCMS: RT = 3.20 min, [MH] - =727.15. 1 H NMR (400MHz, DMSO) δ 12.91 (s, 1H), 11.73 (s, 1H), 10.13 (s, 1H), 9.79 (s, 1H), 8.30 (s, 1H), 8.01 (d, J ＝1.4Hz,1H),7.96(d,J＝2.3Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝9.0,1.8Hz,1H),7.42–7.31(m , 3H), 7.13 (d, J = 7.7 Hz, 2H), 7.07 (d, J = 2.2 Hz, 1H), 6.00 (d, J = 8.0 Hz, 1H), 5.29 (dd, J = 9.8, 5.6 Hz ,1H),3.77(dd,J=20.3,8.9Hz,3H),3.53-3.40(m,4H), 3.20(d,J=10.0Hz,2H), 2.98(s,2H), 1.09(d, J=6.6Hz, 3H).
实施例38Example 38
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-3-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((S)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000218
Figure PCTCN2020130361-appb-000218
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-3-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
Figure PCTCN2020130361-appb-000219
Figure PCTCN2020130361-appb-000219
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和L-缬氨醇(140毫克，1.4毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) N,N-diisopropylethylamine (174 mg, 1.4 mmol) was added dropwise to L-valinol (140 mg, 1.4 mmol) in tetrahydrofuran (3.0 ml), and the dripping was completed, and the reaction was carried out at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到65毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-3-甲基丁-2-基)脲 基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：77.5％)。LCMS:RT＝4.21min,[M-H] -＝897.28。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 65 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((S)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2- Di-tert-butyl dicarboxylate (yield: 77.5%). LCMS: RT = 4.21 min, [MH] - =897.28.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-3-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000220
Figure PCTCN2020130361-appb-000220
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-3-甲基丁-2-基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(65毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-1-hydroxy-3-methylbut-2-yl)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (65 mg, 0.07 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到28毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-3-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：56.1％)。LCMS:RT＝3.29min,[M-H] -＝741.20。 1H NMR(400MHz,DMSO)δ12.91(s,1H),11.72(s,1H),10.14(s,1H),9.80(s,1H),8.45(s,1H),8.01(s,1H),7.96(d,J＝2.2Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.36(dd,J＝17.8,8.6Hz,3H),7.13(d,J＝8.4Hz,2H),7.07(d,J＝2.3Hz,1H),5.98(d,J＝9.2Hz,1H),5.28(dd,J＝10.5,5.9Hz,1H),3.59–3.41(m,3H),3.33(s,3H),3.22(dd,J＝15.2,1.4Hz,2H),3.01(dd,J＝9.9,7.2Hz,2H),0.88(dd,J＝11.8,6.8Hz,6H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 28 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-((S)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H -Indole-2-carboxylic acid (yield: 56.1%). LCMS: RT = 3.29 min, [MH] - =741.20. 1 H NMR (400MHz, DMSO) δ 12.91 (s, 1H), 11.72 (s, 1H), 10.14 (s, 1H), 9.80 (s, 1H), 8.45 (s, 1H), 8.01 (s, 1H) ), 7.96 (d, J = 2.2 Hz, 1H), 7.83 (d, J = 8.6 Hz, 1H), 7.77 (dd, J = 8.6, 2.3 Hz, 1H), 7.36 (dd, J = 17.8, 8.6 Hz , 3H), 7.13 (d, J = 8.4 Hz, 2H), 7.07 (d, J = 2.3 Hz, 1H), 5.98 (d, J = 9.2 Hz, 1H), 5.28 (dd, J = 10.5, 5.9 Hz ,1H),3.59–3.41(m,3H),3.33(s,3H),3.22(dd,J＝15.2,1.4Hz,2H),3.01(dd,J＝9.9,7.2Hz,2H),0.88( dd, J=11.8, 6.8 Hz, 6H).
实施例39Example 39
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-1-羟基-3-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((R)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000221
Figure PCTCN2020130361-appb-000221
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-1-羟基-3-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
Figure PCTCN2020130361-appb-000222
Figure PCTCN2020130361-appb-000222
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和D-缬氨醇(140毫克，1.4毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) N,N-diisopropylethylamine (174 mg, 1.4 mmol) was added dropwise to D-valinol (140 mg, 1.4 mmol) in tetrahydrofuran (3.0 ml), and the dripping was completed, and the reaction was carried out at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到62毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-1-羟基-3-甲基丁-2-基)脲 基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：73.9％)。LCMS:RT＝4.21min,[M-H] -＝897.25。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 62 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((R)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2- Di-tert-butyl dicarboxylate (yield: 73.9%). LCMS: RT = 4.21 min, [MH] - =897.25.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-1-羟基-3-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000223
Figure PCTCN2020130361-appb-000223
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-1-羟基-3-甲基丁-2-基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(62毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-1-hydroxy-3-methylbut-2-yl)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (62 mg, 0.07 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到19毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-1-羟基-3-甲基丁-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：38.1％)。LCMS:RT＝3.30min,[M-H] -＝741.16。 1H NMR(400MHz,DMSO)δ12.93(s,1H),11.72(s,1H),10.13(s,1H),9.80(s,1H),8.44(s,1H),8.01(s,1H),7.96(d,J＝2.2Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.36(dd,J＝18.0,8.5Hz,3H),7.13(d,J＝8.2Hz,2H),7.07(d,J＝1.6Hz,1H),5.98(d,J＝7.9Hz,1H),5.28(dd,J＝10.0,5.1Hz,1H),3.44(dd,J＝8.6,4.1Hz,3H),3.35(dd,J＝12.0,6.7Hz,3H),3.26–3.16(m,2H),3.08–2.95(m,2H),0.88(dd,J＝11.9,6.8Hz,6H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 19 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-((R)-1-hydroxy-3-methylbut-2-yl)ureido)phenyl)propionamido)-1H -Indole-2-carboxylic acid (yield: 38.1%). LCMS: RT = 3.30 min, [MH] - =741.16. 1 H NMR (400MHz, DMSO) δ 12.93 (s, 1H), 11.72 (s, 1H), 10.13 (s, 1H), 9.80 (s, 1H), 8.44 (s, 1H), 8.01 (s, 1H) ), 7.96 (d, J = 2.2 Hz, 1H), 7.83 (d, J = 8.6 Hz, 1H), 7.77 (dd, J = 8.6, 2.3 Hz, 1H), 7.36 (dd, J = 18.0, 8.5 Hz , 3H), 7.13 (d, J = 8.2 Hz, 2H), 7.07 (d, J = 1.6 Hz, 1H), 5.98 (d, J = 7.9 Hz, 1H), 5.28 (dd, J = 10.0, 5.1 Hz ,1H),3.44(dd,J=8.6,4.1Hz,3H), 3.35(dd,J=12.0,6.7Hz,3H), 3.26–3.16(m,2H),3.08–2.95(m,2H), 0.88 (dd, J=11.9, 6.8 Hz, 6H).
实施例40Example 40
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((3-甲基氧杂环丁烷-3-基)甲基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((3-Methyloxetan-3-yl)methyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000224
Figure PCTCN2020130361-appb-000224
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((3-甲基氧杂环丁烷-3-基)甲基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((3-Methyloxetan-3-yl)methyl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
Figure PCTCN2020130361-appb-000225
Figure PCTCN2020130361-appb-000225
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和3-甲基-3-胺甲基-1-氧杂环丁烷(136毫克，1.4毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) And 3-methyl-3-aminomethyl-1-oxetane (136 mg, 1.4 mmol) in tetrahydrofuran (3.0 ml) was added dropwise N,N-diisopropylethylamine (174 mg, 1.4 mmol), after dripping, react at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到 75毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((3-甲基氧杂环丁烷-3-基)甲基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：59.8％)。LCMS:RT＝4.20min,[M+H] +＝897.39。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 75 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((3-methyloxetan-3-yl)methyl)ureido)phenyl)propionamido)-1H-indole-1,2- Di-tert-butyl dicarboxylate (yield: 59.8%). LCMS: RT = 4.20 min, [M+H] + =897.39.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((3-甲基氧杂环丁烷-3-基)甲基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((3-Methyloxetan-3-yl)methyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000226
Figure PCTCN2020130361-appb-000226
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((R)-1-羟基-3-甲基丁-2-基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(75毫克，0.08毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((R)-1-hydroxy-3-methylbut-2-yl)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (75 mg, 0.08 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到32毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((3-甲基氧杂环丁烷-3-基)甲基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：54.0％)。LCMS:RT＝2.75min,[M+H] +＝740.95。 1H NMR(400MHz,DMSO)δ12.96(s,1H),11.75(s,1H),10.85(s,1H),10.15(s,1H),9.79(s,1H),9.34(s,1H),8.01(d,J＝1.2Hz,1H),7.95(d,J＝2.2Hz,1H),7.84(d,J＝8.7Hz,1H),7.79(dd,J＝8.6,2.2Hz,1H),7.37(dt,J＝10.8,5.4Hz,3H),7.28(d,J＝8.4Hz,2H),7.08(d,J＝1.5Hz,1H),5.35(s,1H),5.13(s,1H),4.47(d,J＝9.1Hz,1H),4.33(d,J＝11.9Hz,1H),3.42–3.26(m,5H),3.12(t,J＝11.2Hz,2H),1.00(s,3H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 32 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-((3-methyloxetan-3-yl)methyl)ureido)phenyl)propionamido)-1H -Indole-2-carboxylic acid (yield: 54.0%). LCMS: RT = 2.75 min, [M+H] + =740.95. 1 H NMR (400MHz, DMSO) δ 12.96 (s, 1H), 11.75 (s, 1H), 10.85 (s, 1H), 10.15 (s, 1H), 9.79 (s, 1H), 9.34 (s, 1H) ), 8.01 (d, J = 1.2 Hz, 1H), 7.95 (d, J = 2.2 Hz, 1H), 7.84 (d, J = 8.7 Hz, 1H), 7.79 (dd, J = 8.6, 2.2 Hz, 1H) ), 7.37 (dt, J = 10.8, 5.4 Hz, 3H), 7.28 (d, J = 8.4 Hz, 2H), 7.08 (d, J = 1.5 Hz, 1H), 5.35 (s, 1H), 5.13 (s ,1H),4.47(d,J＝9.1Hz,1H),4.33(d,J＝11.9Hz,1H),3.42-3.26(m,5H),3.12(t,J＝11.2Hz,2H),1.00 (s,3H).
实施例41Example 41
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟甲基)吗啉-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((R)-2-(hydroxymethyl)morpholine-4-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000227
Figure PCTCN2020130361-appb-000227
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟基甲基)吗啉-4-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R)-2-(Hydroxymethyl)morpholine-4-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid Butyl
Figure PCTCN2020130361-appb-000228
Figure PCTCN2020130361-appb-000228
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和(R)-2-吗啉甲醇盐酸盐(208毫克，1.4毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) And (R)-2-morpholine methanol hydrochloride (208 mg, 1.4 mmol) in tetrahydrofuran (3.0 ml) was added dropwise N,N-diisopropylethylamine (174 mg, 1.4 mmol), dropwise After that, react at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2 次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到65毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟基甲基)吗啉-4-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：52.0％)。LCMS:RT＝4.11min,[M-H] -＝911.35。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 65 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((R)-2-(hydroxymethyl)morpholine-4-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (yield: 52.0%). LCMS: RT = 4.11 min, [MH] - = 911.35.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟甲基)吗啉-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R)-2-(hydroxymethyl)morpholine-4-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000229
Figure PCTCN2020130361-appb-000229
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟基甲基)吗啉-4-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(65毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R)-2-(Hydroxymethyl)morpholine-4-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid Butyl ester (65 mg, 0.07 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到25毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((3-甲基氧杂环丁烷-3-基)甲基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：47.2％)。LCMS:RT＝3.14min,[M+H] +＝757.17。 1H NMR(400MHz,DMSO)δ12.87(s,1H),11.71(s,1H),10.13(s,1H),9.78(s,1H),8.52(s,1H),8.03–7.98(m,1H),7.95(d,J＝2.3Hz,1H),7.82(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.3Hz,1H),7.40(dd,J＝14.8,8.7Hz,3H),7.32(dd,J＝8.9,1.9Hz,1H),7.15(d,J＝8.3Hz,1H),7.06(d,J＝1.5Hz,1H),5.30(dd,J＝9.9,5.7Hz,1H),4.05(d,J＝12.7Hz,2H),3.92(d,J＝13.0Hz,2H),3.85(ddd,J＝10.3,1.8,0.9Hz,2H),3.38(dd,J＝8.9,5.8Hz,4H),3.23(dd,J＝14.7,3.0Hz,2H),3.01(dd,J＝16.3,9.3Hz,1H),2.88(td,J＝12.6,3.1Hz,1H),2.68–2.56(m,1H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 25 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-((3-methyloxetan-3-yl)methyl)ureido)phenyl)propionamido)-1H -Indole-2-carboxylic acid (yield: 47.2%). LCMS: RT = 3.14 min, [M+H] + =757.17. 1 H NMR (400MHz, DMSO) δ 12.87 (s, 1H), 11.71 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.52 (s, 1H), 8.03-7.98 (m , 1H), 7.95 (d, J = 2.3 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.76 (dd, J = 8.6, 2.3 Hz, 1H), 7.40 (dd, J = 14.8, 8.7Hz, 3H), 7.32 (dd, J = 8.9, 1.9 Hz, 1H), 7.15 (d, J = 8.3 Hz, 1H), 7.06 (d, J = 1.5 Hz, 1H), 5.30 (dd, J = 9.9, 5.7 Hz, 1H), 4.05 (d, J = 12.7 Hz, 2H), 3.92 (d, J = 13.0 Hz, 2H), 3.85 (ddd, J = 10.3, 1.8, 0.9 Hz, 2H), 3.38 ( dd, J = 8.9, 5.8 Hz, 4H), 3.23 (dd, J = 14.7, 3.0 Hz, 2H), 3.01 (dd, J = 16.3, 9.3 Hz, 1H), 2.88 (td, J = 12.6, 3.1 Hz ,1H),2.68–2.56(m,1H).
实施例42Example 42
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吗啉-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((S)-2-(hydroxymethyl)morpholine-4-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000230
Figure PCTCN2020130361-appb-000230
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟基甲基)吗啉-4-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S)-2-(Hydroxymethyl)morpholine-4-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid Butyl
Figure PCTCN2020130361-appb-000231
Figure PCTCN2020130361-appb-000231
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和(S)-2-吗啉甲醇盐酸盐盐酸盐(208毫克，1.4毫摩尔)的 四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) And (S)-2-morpholine methanol hydrochloride hydrochloride (208 mg, 1.4 mmol) in tetrahydrofuran (3.0 ml) was added dropwise N,N-diisopropylethylamine (174 mg, 1.4 mmol) ), after dripping, react overnight at 65 degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到60毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟基甲基)吗啉-4-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：47.0％)。LCMS:RT＝4.10min,[M-H] -＝911.41。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 60 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((S)-2-(hydroxymethyl)morpholine-4-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (yield: 47.0%). LCMS: RT = 4.10 min, [MH] - = 911.41.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吗啉-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S)-2-(hydroxymethyl)morpholine-4-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000232
Figure PCTCN2020130361-appb-000232
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟基甲基)吗啉-4-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(60毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S)-2-(Hydroxymethyl)morpholine-4-carboxamido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid Butyl ester (60 mg, 0.07 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到30毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吗啉-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：56.6％)。LCMS:RT＝3.15min,[M-H] -＝755.12。 1H NMR(400MHz,DMSO)δ12.95(s,1H),11.73(s,1H),10.14(s,1H),9.80(s,1H),8.53(s,1H),8.01(s,1H),7.96(d,J＝2.2Hz,1H),7.83(d,J＝8.7Hz,1H),7.77(dd,J＝8.6,2.2Hz,1H),7.40(dd,J＝14.9,8.7Hz,3H),7.33(dd,J＝8.9,1.7Hz,1H),7.16(d,J＝7.8Hz,2H),7.07(d,J＝1.5Hz,1H),5.31(dd,J＝10.2,5.2Hz,1H),4.06(d,J＝13.8Hz,2H),3.98–3.81(m,3H),3.80–3.68(m,2H),3.43–3.34(m,4H),3.24(dd,J＝15.6,7.0Hz,1H),3.09–2.96(m,1H),2.88(dd,J＝17.0,6.9Hz,1H),2.74–2.57(m,1H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 30 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-((S)-2-(hydroxymethyl)morpholine-4-carboxamido)phenyl)propionamido)-1H-indole- 2-carboxylic acid (yield: 56.6%). LCMS: RT = 3.15 min, [MH] - =755.12. 1 H NMR (400MHz, DMSO) δ 12.95 (s, 1H), 11.73 (s, 1H), 10.14 (s, 1H), 9.80 (s, 1H), 8.53 (s, 1H), 8.01 (s, 1H) ), 7.96 (d, J = 2.2 Hz, 1H), 7.83 (d, J = 8.7 Hz, 1H), 7.77 (dd, J = 8.6, 2.2 Hz, 1H), 7.40 (dd, J = 14.9, 8.7 Hz , 3H), 7.33 (dd, J = 8.9, 1.7 Hz, 1H), 7.16 (d, J = 7.8 Hz, 2H), 7.07 (d, J = 1.5 Hz, 1H), 5.31 (dd, J = 10.2, 5.2Hz,1H),4.06(d,J=13.8Hz,2H),3.98–3.81(m,3H), 3.80–3.68(m,2H), 3.43–3.34(m,4H), 3.24(dd,J = 15.6, 7.0 Hz, 1H), 3.09–2.96 (m, 1H), 2.88 (dd, J = 17.0, 6.9 Hz, 1H), 2.74–2.57 (m, 1H).
实施例43Example 43
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4S)-4-羟基环己基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((1R,4S)-4-hydroxycyclohexyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000233
Figure PCTCN2020130361-appb-000233
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4S)-4-羟基环己基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((1R,4S)-4-hydroxycyclohexyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000234
Figure PCTCN2020130361-appb-000234
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨 基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和反式-4-氨基环己醇(155毫克，1.4毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) And trans-4-aminocyclohexanol (155 mg, 1.4 mmol) in tetrahydrofuran (3.0 ml) was added dropwise N,N-diisopropylethylamine (174 mg, 1.4 mmol), dripping finished, 65 React overnight in degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到35毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4S)-4-羟基环己基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：27.4％)。LCMS:RT＝4.12min,[M-H] -＝909.22。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 35 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((1R,4S)-4-hydroxycyclohexyl)ureido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl Esters (yield: 27.4%). LCMS: RT = 4.12 min, [MH] - = 909.22.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4S)-4-羟基环己基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((1R,4S)-4-hydroxycyclohexyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000235
Figure PCTCN2020130361-appb-000235
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4S)-4-羟基环己基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(35毫克，0.04毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((1R,4S)-4-hydroxycyclohexyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (35 mg, 0.04 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到15毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4S)-4-羟基环己基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：49.6％)。LCMS:RT＝3.18min,[M+H] +＝755.14。 1H NMR(400MHz,DMSO)δ12.93(s,1H),11.72(s,1H),10.13(s,1H),9.79(s,1H),8.25(s,1H),8.01(d,J＝1.6Hz,1H),7.95(d,J＝2.2Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.44–7.31(m,4H),7.13(d,J＝7.7Hz,2H),7.07(d,J＝1.8Hz,1H),5.99(d,J＝7.7Hz,1H),5.28(dd,J＝10.2,5.0Hz,1H),3.87–3.55(m,3H),3.21(d,J＝12.8Hz,2H),3.08–2.93(m,1H),1.82(t,J＝14.3Hz,4H),1.20(dt,J＝21.9,11.2Hz,4H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 15 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-((1R,4S)-4-hydroxycyclohexyl)ureido)phenyl)propionamido)-1H-indole-2- Carboxylic acid (yield: 49.6%). LCMS: RT = 3.18 min, [M+H] + =755.14. 1 H NMR (400MHz, DMSO) δ 12.93 (s, 1H), 11.72 (s, 1H), 10.13 (s, 1H), 9.79 (s, 1H), 8.25 (s, 1H), 8.01 (d, J ＝1.6Hz,1H),7.95(d,J＝2.2Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.44-7.31(m , 4H), 7.13 (d, J = 7.7 Hz, 2H), 7.07 (d, J = 1.8 Hz, 1H), 5.99 (d, J = 7.7 Hz, 1H), 5.28 (dd, J = 10.2, 5.0 Hz ,1H),3.87–3.55(m,3H),3.21(d,J=12.8Hz,2H),3.08–2.93(m,1H),1.82(t,J=14.3Hz,4H),1.20(dt, J = 21.9, 11.2 Hz, 4H).
实施例44Example 44
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S,3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid
Figure PCTCN2020130361-appb-000236
Figure PCTCN2020130361-appb-000236
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-硝基苯并呋喃-2-羧酸叔丁酯Step A: Synthesis of tert-butyl 5-nitrobenzofuran-2-carboxylate
Figure PCTCN2020130361-appb-000237
Figure PCTCN2020130361-appb-000237
室温下，将5-硝基苯并呋喃-2-甲酸(1.8克，8.7毫摩尔)和二碳酸二叔丁酯(4.8克，22.0毫摩尔)加入N,N-二甲基甲酰胺(20毫升)中，冰浴下，分批加入4-二甲氨基吡啶(0.54克，4.4毫摩尔)，N 2保护下，室温反应过夜。 At room temperature, add 5-nitrobenzofuran-2-carboxylic acid (1.8 g, 8.7 mmol) and di-tert-butyl dicarbonate (4.8 g, 22.0 mmol) to N,N-dimethylformamide (20 (Ml), under ice bath, add 4-dimethylaminopyridine (0.54 g, 4.4 mmol) in batches , and react under N 2 protection at room temperature overnight.
反应结束，加水淬灭，加入100毫升乙酸乙酯，垫硅藻土抽滤，滤饼用50毫升乙酸乙酯洗涤，滤液用乙酸乙酯(100毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(100毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩得到1.9克淡黄色固体5-硝基苯并呋喃-2-羧酸叔丁酯，无需纯化，直接用于下一步反应。LCMS:RT＝4.34min。After the reaction is over, add water to quench, add 100 ml of ethyl acetate, filter with Celite pad, wash the filter cake with 50 ml of ethyl acetate, and extract the filtrate with ethyl acetate (100 ml×3 times). Combine the organic phases. The phase was washed with saturated brine (100 ml×2 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure to obtain 1.9 g of light yellow solid 5-nitrobenzofuran-2-carboxylic acid tert-butyl ester. Purified and used directly in the next reaction. LCMS: RT = 4.34 min.
步骤B：合成5-氨基苯并呋喃-2-羧酸叔丁酯Step B: Synthesis of tert-butyl 5-aminobenzofuran-2-carboxylate
Figure PCTCN2020130361-appb-000238
Figure PCTCN2020130361-appb-000238
室温下，将5-硝基苯并呋喃-2-羧酸叔丁酯(1.9克，7.2毫摩尔l)溶于乙酸乙酯中，加入钯碳(0.5克)，安装氢气球，室温反应9小时。At room temperature, dissolve 5-nitrobenzofuran-2-carboxylic acid tert-butyl ester (1.9 g, 7.2 mmol l) in ethyl acetate, add palladium on carbon (0.5 g), install a hydrogen balloon, and react at room temperature 9 hour.
反应结束，过滤钯碳，滤饼用甲醇洗涤，滤液减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到1.4克棕色固体5-氨基苯并呋喃-2-羧酸叔丁酯(收率：83.4％)。LCMS:RT＝3.05min,[M+H] +＝234.11。 After the reaction, the palladium carbon was filtered, the filter cake was washed with methanol, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). 1.4 g of tert-butyl 5-aminobenzofuran-2-carboxylate was obtained as a brown solid (yield: 83.4%). LCMS: RT=3.05min, [M+H] + =234.11.
步骤C：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-硝基苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯Step C: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-nitrophenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000239
Figure PCTCN2020130361-appb-000239
室温下，将(S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-硝基苯基)丙酸(350毫克，0.7毫摩尔)、5-氨基苯并呋喃-2-羧酸叔丁酯(185毫克，0.8毫摩尔)和2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(411毫克，1.1毫摩尔)加入N,N-二甲基甲酰胺(5.0毫升)中，滴加N,N-二异丙基乙胺(280毫克，2.2毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, (S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-nitrophenyl)propionic acid (350 mg, 0.7 mmol), tert-butyl 5-aminobenzofuran-2-carboxylate (185 mg, 0.8 mmol) and 2-(7-oxybenzene Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (411 mg, 1.1 mmol) was added to N,N-dimethylformamide (5.0 mL) and added dropwise N,N-Diisopropylethylamine (280 mg, 2.2 mmol) was dripped and reacted overnight at room temperature under N 2 protection.
反应结束，加水淬灭，混合液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到400毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-硝基苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(收率：41.1％)。LCMS:RT＝4.15min,[M+H] +＝699.11。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (30 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 400 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- (Yl)-3-(-4-nitrophenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester (yield: 41.1%). LCMS: RT = 4.15 min, [M+H] + =699.11.
步骤D：合成(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯Step D: Synthesis of (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3 -Dioxopiperazin-1-yl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000240
Figure PCTCN2020130361-appb-000240
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-硝基苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(400毫克，0.57毫摩尔)和还原铁粉(320毫克，5.7毫摩尔)加入甲醇(15.0毫升)中，滴加冰醋酸(1.0毫升)，65摄氏度反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-nitrophenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester (400 mg, 0.57 mmol) and reduced iron powder (320 mg, 5.7 mmol) were added to methanol (15.0 ml), glacial acetic acid (1.0 ml) was added dropwise, and reacted at 65 degrees Celsius for 3 hours.
反应结束，向反应液中加入10毫升乙酸乙酯稀释，垫硅藻土抽滤，滤饼用乙酸乙酯洗涤，滤液用饱和碳酸氢钠调pH至10，混合液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到314毫克黄色固体(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(收率：82.1％)。LCMS:RT＝3.58min,[M+H] +＝671.24。 At the end of the reaction, the reaction solution was diluted by adding 10 ml of ethyl acetate, filtered through a pad of diatomaceous earth, the filter cake was washed with ethyl acetate, the filtrate was adjusted to pH 10 with saturated sodium bicarbonate, and the mixture was diluted with ethyl acetate (30 ml × 3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml × 2 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 314 mg of yellow solid (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2, 3-dioxopiperazin-1-yl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester (yield: 82.1%). LCMS: RT = 3.58 min, [M+H] + =671.24.
步骤E：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯Step E: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000241
Figure PCTCN2020130361-appb-000241
N 2保护、冰浴下，向含有(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(314毫克，0.47毫摩尔)四氢呋喃(6.0毫升)中滴加氯甲酸苯酯(117微升)，滴毕，室温反应1小时。 Under the protection of N 2 and ice bath, it will contain (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)benzene (Yl)-2,3-dioxopiperazin-1-yl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester (314mg, 0.47mmol) tetrahydrofuran (6.0ml) was added dropwise chloroformic acid Phenyl ester (117 microliters), after dropping, react at room temperature for 1 hour.
反应结束，加水淬灭，混合液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(20毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/二氯甲烷＝1/1)。得到147毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(收率：39.5％)。LCMS:RT＝4.20min,[M-H] -＝789.21。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (30 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/dichloromethane=1/1). Obtain 147 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester (yield: 39.5%). LCMS: RT = 4.20 min, [MH] - = 789.21.
步骤F：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S,3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯Step F: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid tert Butyl
Figure PCTCN2020130361-appb-000242
Figure PCTCN2020130361-appb-000242
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(80毫克，0.10毫摩尔)和L-异亮氨醇(120毫克，1.0毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(131毫克，1.0毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester (80 mg, 0.10 mmol) and L-isoleucine N,N-diisopropylethylamine (131 mg, 1.0 mmol) was added dropwise to alcohol (120 mg, 1.0 mmol) in tetrahydrofuran (3.0 mL), the dripping was completed, and the reaction was carried out at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/4)。得到45毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S,3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(收率：55.3％)。LCMS:RT＝3.98min,[M+H] +＝814.16。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/4). Obtain 45 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid Tert-butyl ester (yield: 55.3%). LCMS: RT = 3.98 min, [M+H] + =814.16.
步骤G：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S,3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸Step G: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid
Figure PCTCN2020130361-appb-000243
Figure PCTCN2020130361-appb-000243
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S,3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(45毫克，0.05毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid tert Butyl ester (45 mg, 0.05 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到10毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S,3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸(收率：24.1％)。LCMS:RT＝3.58min,[M-H] -＝756.32。 1H NMR(500MHz,DMSO)δ13.57(s,1H),10.38(s,1H),9.81(s,1H),8.47(s,1H),8.19(d,J＝1.9Hz,1H),7.97(d,J＝2.2Hz,1H),7.84(d,J＝8.6Hz,1H),7.78(dd,J＝8.6,2.3Hz,1H),7.70(s,1H),7.68(d,J＝9.0Hz,1H),7.59–7.54(m,2H),7.35(d,J＝8.4Hz,2H),7.14(d,J＝7.2Hz,2H),6.02(d,J＝8.5Hz,1H),5.30(dd,J＝10.0,5.6Hz,1H),3.59–3.45(m,3H),3.41(dd,J＝10.5,4.5Hz,3H),3.28–3.14(m,2H),3.02(dd,J＝12.6,9.5Hz,1H),1.68–1.56(m,1H),1.56–1.42(m,1H),1.08(dt,J＝23.4,9.1Hz,1H),0.87(t,J＝6.7Hz,6H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 10 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido) Benzofuran-2-carboxylic acid (yield: 24.1%). LCMS: RT = 3.58 min, [MH] - = 756.32. 1 H NMR (500MHz, DMSO) δ 13.57 (s, 1H), 10.38 (s, 1H), 9.81 (s, 1H), 8.47 (s, 1H), 8.19 (d, J = 1.9 Hz, 1H), 7.97 (d, J = 2.2 Hz, 1H), 7.84 (d, J = 8.6 Hz, 1H), 7.78 (dd, J = 8.6, 2.3 Hz, 1H), 7.70 (s, 1H), 7.68 (d, J ＝9.0Hz,1H),7.59–7.54(m,2H),7.35(d,J＝8.4Hz,2H),7.14(d,J＝7.2Hz,2H),6.02(d,J＝8.5Hz,1H ), 5.30(dd,J=10.0,5.6Hz,1H),3.59–3.45(m,3H),3.41(dd,J=10.5,4.5Hz,3H), 3.28–3.14(m,2H),3.02( dd,J＝12.6,9.5Hz,1H),1.68–1.56(m,1H),1.56–1.42(m,1H),1.08(dt,J＝23.4,9.1Hz,1H),0.87(t,J＝ 6.7Hz, 6H).
实施例45Example 45
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-(Hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000244
Figure PCTCN2020130361-appb-000244
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(Hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000245
Figure PCTCN2020130361-appb-000245
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(200毫克，0.22毫摩尔)和3-羟甲基哌啶(260毫克，2.2毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(290毫克，2.2毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (200 mg, 0.22 mmol) And 3-hydroxymethylpiperidine (260 mg, 2.2 mmol) in tetrahydrofuran (5.0 mL) was added dropwise N,N-diisopropylethylamine (290 mg, 2.2 mmol), after dripping, react at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到80毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：39.9％)。LCMS:RT＝3.74min,[M-H] -＝909.22。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 80 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(3-(hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester ( Yield: 39.9%). LCMS: RT = 3.74 min, [MH] -= 909.22.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000246
Figure PCTCN2020130361-appb-000246
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(80毫克，0.09毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(Hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (80 Mg, 0.09 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到27毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：39.7％)。LCMS:RT＝3.28min,[M+H] +＝755.10。 1H NMR(500MHz,DMSO)δ12.94(s,1H),11.74(s,1H),10.17(s,1H),9.82(s,1H),8.43(s,1H),8.02(s,1H),7.97(d,J＝2.3Hz,1H),7.84(d,J＝8.6Hz,1H),7.78(dd,J＝8.6,2.3Hz,1H),7.40(t,J＝9.1Hz,3H),7.34(dd,J＝8.9,1.9Hz,1H),7.15(d,J＝8.0Hz,2H),7.08(d,J＝1.5Hz,1H),5.31(dd,J＝9.8,5.6Hz,1H),4.08(d,J＝13.9Hz,2H),3.97(d,J＝10.5Hz,2H),3.39–3.19(m,4H),3.08–2.91(m,2H),2.81(t,J＝13.1Hz,1H),2.12–1.94(m,1H),1.74(d,J＝12.0Hz,1H),1.63(d,J＝12.5Hz,1H),1.55(s,1H),1.40(d,J＝16.4Hz,1H),1.23–1.11(m,1H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 27 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-(hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (Yield: 39.7%). LCMS: RT = 3.28 min, [M+H] + =755.10. 1 H NMR (500MHz, DMSO) δ 12.94 (s, 1H), 11.74 (s, 1H), 10.17 (s, 1H), 9.82 (s, 1H), 8.43 (s, 1H), 8.02 (s, 1H) ), 7.97 (d, J = 2.3 Hz, 1H), 7.84 (d, J = 8.6 Hz, 1H), 7.78 (dd, J = 8.6, 2.3 Hz, 1H), 7.40 (t, J = 9.1 Hz, 3H ), 7.34 (dd, J = 8.9, 1.9 Hz, 1H), 7.15 (d, J = 8.0 Hz, 2H), 7.08 (d, J = 1.5 Hz, 1H), 5.31 (dd, J = 9.8, 5.6 Hz ,1H),4.08(d,J=13.9Hz,2H),3.97(d,J=10.5Hz,2H),3.39–3.19(m,4H),3.08–2.91(m,2H),2.81(t, J = 13.1Hz, 1H), 2.12-1.94 (m, 1H), 1.74 (d, J = 12.0 Hz, 1H), 1.63 (d, J = 12.5 Hz, 1H), 1.55 (s, 1H), 1.40 ( d, J = 16.4 Hz, 1H), 1.23-1.11 (m, 1H).
实施例46Example 46
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid
Figure PCTCN2020130361-appb-000247
Figure PCTCN2020130361-appb-000247
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester
Figure PCTCN2020130361-appb-000248
Figure PCTCN2020130361-appb-000248
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(75毫克，0.09毫摩尔)和3-氨基-3-甲基-1-丁醇(98毫克，0.9毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(123毫克，0.9毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester (75 mg, 0.09 mmol) and 3-amino-3 -Methyl-1-butanol (98 mg, 0.9 mmol) in tetrahydrofuran (3.0 mL) was added dropwise N,N-diisopropylethylamine (123 mg, 0.9 mmol), after dripping, react at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/4)。得到35毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(收率：48.6％)。LCMS:RT＝3.90min,[M+H] +＝798.28。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/4). Obtain 35 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester (yield : 48.6%). LCMS: RT = 3.90 min, [M+H] + =798.28.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid
Figure PCTCN2020130361-appb-000249
Figure PCTCN2020130361-appb-000249
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸叔丁酯(35毫克，0.04毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)benzofuran-2-carboxylic acid tert-butyl ester (35 mg, 0.04 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到7毫克白色固5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(4-羟基-2-甲基丁-2-基)脲基)苯基)丙酰胺基)苯并呋喃-2-羧酸(收率：23.5％)。LCMS:RT＝3.45min,[M-H] -＝742.12。 1H NMR(500MHz,DMSO)δ13.57(s,1H),10.38(s,1H),9.81(s,1H),8.31(s,1H),8.19(d,J＝1.8Hz,1H),7.97(d,J＝2.2Hz,1H),7.84(d,J＝8.6Hz,1H),7.78(dd,J＝8.6,2.3Hz,1H),7.68(d,J＝9.2Hz,2H),7.57(d,J＝10.9Hz,1H),7.32(d,J＝8.5Hz,2H),7.13(d,J＝6.5Hz,2H),6.01(s,1H),5.30(dd,J＝10.0,5.7Hz,1H),4.39(t,J＝5.0Hz,1H),3.51(dd,J＝11.8,7.2Hz,2H),3.23(dd,J＝13.7,4.9Hz,1H),3.11–2.98(m,1H),2.02(dd,J＝17.3,9.6Hz,1H),1.82(t,J＝7.3Hz,2H),1.55–1.42(m,1H),1.28(s,6H),0.87(t,J＝6.4Hz,1H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 7 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(3-(4-hydroxy-2-methylbut-2-yl)ureido)phenyl)propionamido)benzofuran-2- Carboxylic acid (yield: 23.5%). LCMS: RT = 3.45 min, [MH] - = 742.12. 1 H NMR (500MHz, DMSO) δ 13.57 (s, 1H), 10.38 (s, 1H), 9.81 (s, 1H), 8.31 (s, 1H), 8.19 (d, J = 1.8 Hz, 1H), 7.97 (d, J = 2.2 Hz, 1H), 7.84 (d, J = 8.6 Hz, 1H), 7.78 (dd, J = 8.6, 2.3 Hz, 1H), 7.68 (d, J = 9.2 Hz, 2H), 7.57(d,J＝10.9Hz,1H),7.32(d,J＝8.5Hz,2H),7.13(d,J＝6.5Hz,2H),6.01(s,1H),5.30(dd,J＝10.0 ,5.7Hz,1H), 4.39(t,J=5.0Hz,1H),3.51(dd,J=11.8,7.2Hz,2H), 3.23(dd,J=13.7,4.9Hz,1H),3.11–2.98 (m,1H),2.02(dd,J=17.3,9.6Hz,1H),1.82(t,J=7.3Hz,2H),1.55-1.42(m,1H),1.28(s,6H),0.87( t,J=6.4Hz,1H).
实施例47Example 47
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4R)-4-羟基环己基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((1R,4R)-4-hydroxycyclohexyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000250
Figure PCTCN2020130361-appb-000250
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4R)-4-羟基环己基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((1R,4R)-4-hydroxycyclohexyl)ureido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000251
Figure PCTCN2020130361-appb-000251
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和顺式-4-氨基环己醇(155毫克，1.4毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) And cis-4-aminocyclohexanol (155 mg, 1.4 mmol) in tetrahydrofuran (3.0 ml) was added dropwise N,N-diisopropylethylamine (174 mg, 1.4 mmol), dripping finished, 65 degrees Celsius React overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到60毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4R)-4-羟基环己基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：47.0％)。LCMS:RT＝4.12min,[M-H] -＝909.22。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 60 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((1R,4R)-4-hydroxycyclohexyl)ureido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl Ester (yield: 47.0%). LCMS: RT = 4.12 min, [MH] - = 909.22.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4R)-4-羟基环己基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((1R,4R)-4-hydroxycyclohexyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000252
Figure PCTCN2020130361-appb-000252
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4R)-4-羟基环己基)脲基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(60毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((1R,4R)-4-hydroxycyclohexyl)ureido)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (60 mg, 0.07 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到20毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((1R,4R)-4-羟基环己基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：37.9％)。LCMS:RT＝3.21min,[M+H] +＝755.20。 1H NMR(400MHz,DMSO)δ12.92(s,1H),11.73(s,1H),10.13(s,1H),9.80(s,1H),8.28(s,1H),8.01(d,J＝1.8Hz,1H),7.96(d,J＝2.3Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.39(d,J＝9.0Hz,1H),7.33(d,J＝8.6Hz,2H),7.13(d,J＝9.1Hz,2H),7.07(d,J＝2.0Hz,1H),5.28(dd,J＝9.5,5.6Hz,1H),4.47(s,1H),3.74(s,1H),3.67–3.52(m,2H),3.21(dd,J＝19.3,5.3Hz,2H),3.09–2.88(m,2H),1.77–1.27(m,8H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 20 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiprazin-1-yl)-3-(4-(3-((1R,4R)-4-hydroxycyclohexyl)ureido)phenyl)propionamido)-1H-indole-2- Carboxylic acid (yield: 37.9%). LCMS: RT = 3.21 min, [M+H] + =755.20. 1 H NMR (400MHz, DMSO) δ 12.92 (s, 1H), 11.73 (s, 1H), 10.13 (s, 1H), 9.80 (s, 1H), 8.28 (s, 1H), 8.01 (d, J ＝1.8Hz,1H),7.96(d,J＝2.3Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.39(d,J ＝9.0Hz,1H),7.33(d,J＝8.6Hz,2H), 7.13(d,J＝9.1Hz,2H), 7.07(d,J＝2.0Hz,1H), 5.28(dd,J＝9.5 ,5.6Hz,1H),4.47(s,1H),3.74(s,1H),3.67–3.52(m,2H),3.21(dd,J=19.3,5.3Hz,2H),3.09–2.88(m, 2H), 1.77–1.27 (m, 8H).
实施例48Example 48
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((S)-2-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000253
Figure PCTCN2020130361-appb-000253
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S)-2-(Hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl
Figure PCTCN2020130361-appb-000254
Figure PCTCN2020130361-appb-000254
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和L-脯氨醇(136毫克，0.14毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) N,N-diisopropylethylamine (174 mg, 1.4 mmol) was added dropwise to L-prolinol (136 mg, 0.14 mmol) in tetrahydrofuran (3.0 ml), the dripping was completed, and the reaction was carried out at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到40毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：31.8％)。LCMS:RT＝3.78min,[M-H] -＝895.31。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 40 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((S)-2-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (yield: 31.8%). LCMS: RT = 3.78 min, [MH] - =895.31.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S)-2-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000255
Figure PCTCN2020130361-appb-000255
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(40毫克，0.045毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S)-2-(Hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl ester (40 mg, 0.045 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到6毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：18.0％)。LCMS:RT＝3.28min,[M+H] +＝741.10。 1H NMR(400MHz,DMSO)δ12.94(s,1H),11.73(s,1H),10.16(s,1H),9.80(s, 1H),8.51(s,1H),8.01(s,1H),7.96(d,J＝2.2Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.39(dd,J＝8.5,6.3Hz,3H),7.33(dd,J＝9.2,1.6Hz,1H),7.16(d,J＝8.5Hz,2H),7.07(d,J＝1.8Hz,1H),5.30(dd,J＝10.4,6.1Hz,1H),3.94(s,1H),3.82–3.75(m,2H),3.56(dd,J＝11.6,6.2Hz,2H),3.23(dd,J＝12.3,5.5Hz,1H),3.11–3.06(m,1H),3.06–2.93(m,1H),2.00(dd,J＝15.9,9.1Hz,1H),1.89(dd,J＝18.7,11.5Hz,2H),1.77(dd,J＝14.5,7.3Hz,2H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 6 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-((S)-2-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole- 2-carboxylic acid (yield: 18.0%). LCMS: RT = 3.28 min, [M+H] + =741.10. 1 H NMR (400MHz, DMSO) δ 12.94 (s, 1H), 11.73 (s, 1H), 10.16 (s, 1H), 9.80 (s, 1H), 8.51 (s, 1H), 8.01 (s, 1H) ), 7.96 (d, J = 2.2 Hz, 1H), 7.83 (d, J = 8.6 Hz, 1H), 7.77 (dd, J = 8.6, 2.3 Hz, 1H), 7.39 (dd, J = 8.5, 6.3 Hz , 3H), 7.33 (dd, J = 9.2, 1.6 Hz, 1H), 7.16 (d, J = 8.5 Hz, 2H), 7.07 (d, J = 1.8 Hz, 1H), 5.30 (dd, J = 10.4, 6.1Hz, 1H), 3.94 (s, 1H), 3.82-3.75 (m, 2H), 3.56 (dd, J = 11.6, 6.2 Hz, 2H), 3.23 (dd, J = 12.3, 5.5 Hz, 1H), 3.11–3.06(m,1H),3.06–2.93(m,1H),2.00(dd,J=15.9,9.1Hz,1H),1.89(dd,J=18.7,11.5Hz,2H),1.77(dd, J=14.5, 7.3 Hz, 2H).
实施例49Example 49
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((R)-2-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000256
Figure PCTCN2020130361-appb-000256
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R)-2-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl
Figure PCTCN2020130361-appb-000257
Figure PCTCN2020130361-appb-000257
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(-4-((苯氧基羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(120毫克，0.14毫摩尔)和L-脯氨醇(136毫克，0.14毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(174毫克，1.4毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(-4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.14 mmol) N,N-diisopropylethylamine (174 mg, 1.4 mmol) was added dropwise to L-prolinol (136 mg, 0.14 mmol) in tetrahydrofuran (3.0 ml), the dripping was completed, and the reaction was carried out at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到45毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：35.8％)。LCMS:RT＝3.78min,[M-H] -＝895.25。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (20 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 45 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((R)-2-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (yield: 35.8%). LCMS: RT = 3.78 min, [MH] - =895.25.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R)-2-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000258
Figure PCTCN2020130361-appb-000258
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(45毫克，0.05毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S)-2-(Hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl ester (45 mg, 0.05 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到18毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R)-2-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：54.0％)。LCMS:RT＝3.28min,[M+H] +＝741.13。 1H NMR(400MHz,DMSO)δ13.04(s,1H),11.74(s,1H),10.16(s,1H),9.80(s,1H),8.52(s,1H),8.52(s,1H),8.01(s,1H),7.96(d,J＝2.3Hz,1H),7.83(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.58–7.37(m,3H),7.35(d,J＝3.8Hz,1H),7.16(d,J＝8.3Hz,2H),7.07(d,J＝1.5Hz,1H),5.30(dd,J＝11.3,7.1Hz,1H),3.94(s,2H),3.87–3.63(m,3H),3.23(dd,J＝16.8,2.5Hz,2H),3.03(dd,J＝19.9,8.5Hz,1H),2.00(dd,J＝14.1,7.9Hz,1H),1.88(dd,J＝17.7,11.3Hz,2H),1.80–1.66(m,2H),1.59–1.40(m,1H),0.86(t,J＝5.4Hz,1H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 18 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-((R)-2-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole- 2-carboxylic acid (yield: 54.0%). LCMS: RT = 3.28 min, [M+H] + =741.13. 1 H NMR (400MHz, DMSO) δ 13.04 (s, 1H), 11.74 (s, 1H), 10.16 (s, 1H), 9.80 (s, 1H), 8.52 (s, 1H), 8.52 (s, 1H) ), 8.01 (s, 1H), 7.96 (d, J = 2.3 Hz, 1H), 7.83 (d, J = 8.6 Hz, 1H), 7.77 (dd, J = 8.6, 2.3 Hz, 1H), 7.58–7.37 (m, 3H), 7.35 (d, J = 3.8 Hz, 1H), 7.16 (d, J = 8.3 Hz, 2H), 7.07 (d, J = 1.5 Hz, 1H), 5.30 (dd, J = 11.3, 7.1Hz, 1H), 3.94 (s, 2H), 3.87–3.63 (m, 3H), 3.23 (dd, J = 16.8, 2.5 Hz, 2H), 3.03 (dd, J = 19.9, 8.5 Hz, 1H), 2.00(dd,J=14.1,7.9Hz,1H),1.88(dd,J=17.7,11.3Hz,2H),1.80–1.66(m,2H),1.59–1.40(m,1H),0.86(t, J=5.4Hz, 1H).
实施例50Example 50
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000259
Figure PCTCN2020130361-appb-000259
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000260
Figure PCTCN2020130361-appb-000260
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(200毫克，0.22毫摩尔)和哌啶-4-甲腈(29毫克，0.27毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(67毫克，0.67毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (200 mg, 0.22 mmol) and Piperidine-4-carbonitrile (29 mg, 0.27 mmol) in tetrahydrofuran (5.0 mL) was added dropwise with N,N-diisopropylethylamine (67 mg, 0.67 mmol), after dripping, react at 65 degrees Celsius overnight .
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/10)。得到130毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：64％)。LCMS:RT＝4.26min,[M+H] +＝906.33。 1H NMR(500MHz,DMSO)δ10.35(s,1H),9.79(s,1H),8.53(s,1H),8.05(s,1H),7.95(d,J＝2.1Hz,1H),7.86(d,J＝9.0Hz,1H),7.82(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.1Hz,1H),7.56(dd,J＝9.1,1.8Hz,1H),7.40(d,J＝8.4Hz,2H),7.25(s,1H),7.16(t,J＝10.8Hz,2H),5.31(dd,J＝9.9,5.7Hz,1H),3.69(dd,J＝9.5,5.6Hz,3H),3.30–3.21(m,4H),3.14–2.96(m,3H),1.91–1.85(m,2H),1.79–1.62(m,3H),1.60(s,9H),1.55(s,9H)。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/10). To obtain 130 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(4-cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (Yield: 64%). LCMS: RT = 4.26 min, [M+H] + =906.33. 1 H NMR (500MHz, DMSO) δ 10.35 (s, 1H), 9.79 (s, 1H), 8.53 (s, 1H), 8.05 (s, 1H), 7.95 (d, J = 2.1Hz, 1H), 7.86(d,J=9.0Hz,1H),7.82(d,J=8.6Hz,1H),7.77(dd,J=8.6,2.1Hz,1H),7.56(dd,J=9.1,1.8Hz,1H ), 7.40 (d, J = 8.4 Hz, 2H), 7.25 (s, 1H), 7.16 (t, J = 10.8 Hz, 2H), 5.31 (dd, J = 9.9, 5.7 Hz, 1H), 3.69 (dd ,J=9.5,5.6Hz,3H),3.30–3.21(m,4H),3.14–2.96(m,3H),1.91–1.85(m,2H),1.79–1.62(m,3H),1.60(s ,9H),1.55(s,9H).
实施例51Example 51
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000261
Figure PCTCN2020130361-appb-000261
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000262
Figure PCTCN2020130361-appb-000262
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(93毫克，0.1毫摩尔)加入二氯甲烷(2.5毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (93 mg, 0.1 (Mmol) was added to dichloromethane (2.5 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到30.15毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：39.5％)。LCMS:RT＝3.37min,[M+H] +＝750.08。 1H NMR(500MHz,DMSO)δ12.93(s,1H),11.72(s,1H),10.14(s,1H),9.79(s,1H),8.53(s,1H),8.00(s,1H),7.95(d,J＝2.3Hz,1H),7.82(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.39(dd,J＝12.4,8.8Hz,3H),7.32(dd,J＝8.9,1.6Hz,1H),7.15(d,J＝6.9Hz,2H),7.06(s,1H),5.30(dd,J＝9.8,5.8Hz,1H),3.69(dd,J＝9.3,5.6Hz,4H),3.30–3.15(m,4H),3.15–2.88(m,3H),1.88(dd,J＝8.2,4.5Hz,2H),1.73–1.61(m,2H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 30.15 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (Yield: 39.5%). LCMS: RT = 3.37 min, [M+H] + =750.08. 1 H NMR (500MHz, DMSO) δ 12.93 (s, 1H), 11.72 (s, 1H), 10.14 (s, 1H), 9.79 (s, 1H), 8.53 (s, 1H), 8.00 (s, 1H) ), 7.95 (d, J = 2.3 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.77 (dd, J = 8.6, 2.3 Hz, 1H), 7.39 (dd, J = 12.4, 8.8 Hz ,3H),7.32(dd,J=8.9,1.6Hz,1H),7.15(d,J=6.9Hz,2H),7.06(s,1H),5.30(dd,J=9.8,5.8Hz,1H) , 3.69(dd,J=9.3,5.6Hz,4H),3.30–3.15(m,4H),3.15–2.88(m,3H),1.88(dd,J=8.2,4.5Hz,2H),1.73-1.61 (m, 2H).
实施例52Example 52
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-(二甲基氨基)乙氧基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(2-(dimethylamino)ethoxy)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000263
Figure PCTCN2020130361-appb-000263
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-(二甲基氨基)乙氧基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-(dimethylamino)ethoxy)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
Figure PCTCN2020130361-appb-000264
Figure PCTCN2020130361-appb-000264
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(165毫克，0.18毫摩尔)和N，N-二甲基-2-(哌啶-4-基氧基)乙-1-胺(222毫克，0.56毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(191毫克，1.5毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (165 mg, 0.18 mmol) and N,N-Dimethyl-2-(piperidin-4-yloxy)ethan-1-amine (222 mg, 0.56 mmol) in tetrahydrofuran (5.0 mL) was added dropwise with N,N-diisopropyl Ethylamine (191 mg, 1.5 mmol) was dripped and reacted overnight at 65 degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/9)。得到110毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-(二甲基氨基)乙氧基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：61.5％)。LCMS:RT＝3.41min,[M+H] +＝968.33。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtain 110 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(4-(2-(dimethylamino)ethoxy)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2- Di-tert-butyl dicarboxylate (yield: 61.5%). LCMS: RT=3.41min, [M+H] + =968.33.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-(二甲基氨基)乙氧基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-(dimethylamino)ethoxy)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000265
Figure PCTCN2020130361-appb-000265
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-(二甲基氨基)乙氧基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(110毫克，0.11毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-(dimethylamino)ethoxy)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate (110 mg, 0.11 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到42毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-(二甲基氨基)乙氧基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：43.5％)。LCMS:RT＝2.77min,[M+H] +＝812.41。 1H NMR(500MHz,DMSO)δ12.93(s,1H),11.73(s,1H),10.14(s,1H),9.79(s,1H),9.23(s,1H),8.50(s,1H),8.00(s,1H),7.95(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.3Hz,1H),7.39(dd,J＝12.9,8.8Hz,3H),7.33(dd,J＝8.9,1.7Hz,1H),7.13(t,J＝12.4Hz,2H),7.06(d,J＝1.7Hz,1H),5.30(dd,J＝9.7,5.8Hz,1H),3.80(d,J＝13.5Hz,4H),3.75–3.71(m,3H),3.61–3.56(m,1H),3.31–3.09(m,6H),3.05–2.96(m,1H),2.80(d,J＝4.9Hz,6H),1.87(d,J＝9.7Hz,2H),1.44(d,J＝8.9Hz,2H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 42 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-(4-(2-(dimethylamino)ethoxy)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole- 2-carboxylic acid (yield: 43.5%). LCMS: RT = 2.77 min, [M+H] + =812.41. 1 H NMR (500MHz, DMSO) δ 12.93 (s, 1H), 11.73 (s, 1H), 10.14 (s, 1H), 9.79 (s, 1H), 9.23 (s, 1H), 8.50 (s, 1H) ),8.00(s,1H),7.95(d,J=2.2Hz,1H),7.82(d,J=8.6Hz,1H),7.77(dd,J=8.6,2.3Hz,1H),7.39(dd ,J＝12.9,8.8Hz,3H),7.33(dd,J＝8.9,1.7Hz,1H),7.13(t,J＝12.4Hz,2H),7.06(d,J＝1.7Hz,1H),5.30 (dd,J=9.7,5.8Hz,1H), 3.80(d,J=13.5Hz,4H), 3.75–3.71(m,3H),3.61–3.56(m,1H),3.31–3.09(m,6H ), 3.05-2.96 (m, 1H), 2.80 (d, J=4.9 Hz, 6H), 1.87 (d, J=9.7 Hz, 2H), 1.44 (d, J=8.9 Hz, 2H).
实施例53Example 53
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲氧基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Methoxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000266
Figure PCTCN2020130361-appb-000266
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲氧基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Methoxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000267
Figure PCTCN2020130361-appb-000267
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.17毫摩尔)和4-甲氧基哌啶(39毫克，0.34毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(65毫克，0.50毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.17 mmol) and 4-Methoxypiperidine (39mg, 0.34mmol) in tetrahydrofuran (5.0ml) was added dropwise with N,N-diisopropylethylamine (65mg, 0.50mmol), after dripping, react at 65°C overnight .
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/6)。得到57毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲氧基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：37.2％)。LCMS:RT＝4.29min,[M+H] +＝911.25。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/6). Obtain 57 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(4-methoxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield : 37.2%). LCMS: RT = 4.29 min, [M+H] + =911.25.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲氧基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-methoxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000268
Figure PCTCN2020130361-appb-000268
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲氧基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(57.0毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Methoxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (57.0 mg, 0.07 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到19.1毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲氧基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：40.4％)。LCMS:RT＝3.33min,[M+H] +＝755.15。 1H NMR(400MHz,DMSO)δ12.92(s,1H),11.72(s,1H),10.14(s,1H),9.78(s,1H),8.46(s,1H),8.00(s,1H),7.94(d,J＝2.2Hz,1H),7.78(dt,J＝8.6,5.4Hz,2H),7.49–7.26(m,4H),7.09(dd,J＝32.1,4.9Hz,3H),5.34–5.25(m,1H),3.76(d,J＝13.8Hz,4H),3.25(s,2H),3.09(dd,J＝16.3,6.6Hz,2H),3.05–2.95(m,1H),1.99(dd,J＝14.4,6.8Hz,1H),1.81(s,2H),1.37(dd,J＝11.3,6.7Hz,2H),1.22(s,4H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 19.1 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-(4-methoxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 40.4% ). LCMS: RT = 3.33 min, [M+H] + =755.15. 1 H NMR (400MHz, DMSO) δ 12.92 (s, 1H), 11.72 (s, 1H), 10.14 (s, 1H), 9.78 (s, 1H), 8.46 (s, 1H), 8.00 (s, 1H) ), 7.94 (d, J = 2.2 Hz, 1H), 7.78 (dt, J = 8.6, 5.4 Hz, 2H), 7.49-7.26 (m, 4H), 7.09 (dd, J = 32.1, 4.9 Hz, 3H) ,5.34–5.25(m,1H),3.76(d,J=13.8Hz,4H), 3.25(s,2H), 3.09(dd,J=16.3,6.6Hz,2H),3.05–2.95(m,1H ), 1.99 (dd, J=14.4, 6.8 Hz, 1H), 1.81 (s, 2H), 1.37 (dd, J=11.3, 6.7 Hz, 2H), 1.22 (s, 4H).
实施例54Example 54
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲基哌嗪-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸
Figure PCTCN2020130361-appb-000269
Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-Methylpiperazine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000269
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲基哌嗪-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Methylpiperazine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000270
Figure PCTCN2020130361-appb-000270
将(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.19毫摩尔)，4-甲基哌嗪-1-碳酰氯(47毫克，0.29毫摩尔)溶于二氯甲烷(5.0毫升)中，加入吡啶(61毫克，0.78毫摩尔)。室温反应18小时。Add (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-diox Piperazine-1-yl) propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.19 mmol), 4-methylpiperazine-1-carbonyl chloride ( 47 mg, 0.29 mmol) was dissolved in dichloromethane (5.0 mL), and pyridine (61 mg, 0.78 mmol) was added. React at room temperature for 18 hours.
加水淬灭反应，加乙酸乙酯(20毫升)稀释，水和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏，所得残余物用TLC板纯化得84毫克浅黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲基哌嗪-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：48.3％)。LCMS:RT＝3.47min,[M+H] +＝896.29。 The reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml × 3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by TLC plate to obtain 84 mg of light yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-(4-Methylpiperazine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 48.3%). LCMS: RT = 3.47 min, [M+H] + =896.29.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲基哌嗪-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Methylpiperazine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000271
Figure PCTCN2020130361-appb-000271
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲基哌嗪-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(84毫克，0.09毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-Methylpiperazine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (84 mg, 0.09 (Mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到21毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-甲基哌嗪-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：28.9％)。LCMS:RT＝2.66min,[M+H] +＝740.20。 1H NMR(400MHz,DMSO)δ12.92(s,1H),11.72(s,1H),10.13(s,1H),9.79(s,1H),9.71(s,1H),8.73(s,1H),7.99(s,1H),7.94(d,J＝2.2Hz,1H),7.78(dt,J＝8.6,5.4Hz,2H),7.43–7.30(m,3H),7.26–7.01(m,3H),5.30(dt,J＝10.4,5.4Hz,1H),4.24(d,J＝14.3Hz,2H),3.44(d,J＝11.6Hz,2H),3.22(d,J＝10.7Hz,1H),3.17–3.07(m,2H),2.99(d,J＝10.1Hz,3H),2.81(d,J＝3.7Hz,2H),1.99(dd,J＝14.5,6.9Hz,1H),1.22(s,4H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 21 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-(4-methylpiperazine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 28.9%) . LCMS: RT = 2.66 min, [M+H] + =740.20. 1 H NMR (400MHz, DMSO) δ 12.92 (s, 1H), 11.72 (s, 1H), 10.13 (s, 1H), 9.79 (s, 1H), 9.71 (s, 1H), 8.73 (s, 1H) ),7.99(s,1H),7.94(d,J=2.2Hz,1H),7.78(dt,J=8.6,5.4Hz,2H),7.43-7.30(m,3H),7.26-7.01(m, 3H), 5.30 (dt, J = 10.4, 5.4 Hz, 1H), 4.24 (d, J = 14.3 Hz, 2H), 3.44 (d, J = 11.6 Hz, 2H), 3.22 (d, J = 10.7 Hz, 1H), 3.17–3.07 (m, 2H), 2.99 (d, J = 10.1Hz, 3H), 2.81 (d, J = 3.7 Hz, 2H), 1.99 (dd, J = 14.5, 6.9 Hz, 1H), 1.22(s, 4H).
实施例55Example 55
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((R))-3-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000272
Figure PCTCN2020130361-appb-000272
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R))-3-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000273
Figure PCTCN2020130361-appb-000273
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(300毫克，0.34毫摩尔)和(R)-哌啶-3-醇(92毫克，0.67毫摩尔)的四氢呋喃(10.0毫升)中滴加三乙胺(101毫克，1.01毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (300 mg, 0.34 mmol) and Triethylamine (101 mg, 1.01 mmol) was added dropwise to (R)-piperidin-3-ol (92 mg, 0.67 mmol) in tetrahydrofuran (10.0 ml), and after the dripping was completed, the reaction was carried out at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到233毫克红棕色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：77.1％)。LCMS:RT＝3.69min,[M+H] +＝897.28。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 233 mg of red-brown solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1 -Yl)-3-(4-((R))-3-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl Esters (yield: 77.1%). LCMS: RT = 3.69 min, [M+H] + =897.28.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R))-3-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000274
Figure PCTCN2020130361-appb-000274
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(233毫克，0.29毫摩尔)加入二氯甲烷(10.0毫升)中，滴加三氟乙酸(2.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R))-3-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester ( 233 mg, 0.29 mmol) was added to dichloromethane (10.0 mL), trifluoroacetic acid (2.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％； 检测波长：254nm。纯化后，干燥得到76.82毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-羟基哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：39.9％)。LCMS:RT＝3.19min,[M+H] +＝741.21。 1H NMR(400MHz,DMSO)δ12.91(s,1H),11.72(s,1H),10.13(s,1H),9.78(s,1H),8.41(s,1H),8.00(s,1H),7.94(d,J＝2.2Hz,1H),7.81(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.2Hz,1H),7.39(d,J＝8.6Hz,2H),7.37–7.29(m,2H),7.13(d,J＝8.1Hz,2H),7.05(d,J＝1.6Hz,1H),5.29(dd,J＝9.7,5.7Hz,1H),3.94(d,J＝9.4Hz,2H),3.77(d,J＝13.2Hz,2H),3.26–3.12(m,2H),3.08–2.80(m,3H),2.71–2.63(m,1H),1.75(dd,J＝77.7,8.9Hz,3H),1.40–1.25(m,2H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; Detection wavelength: 254nm. After purification, it was dried to obtain 76.82 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-((R))-3-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield : 39.9%). LCMS: RT = 3.19 min, [M+H] + =741.21. 1 H NMR (400MHz, DMSO) δ 12.91 (s, 1H), 11.72 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.41 (s, 1H), 8.00 (s, 1H) ), 7.94 (d, J = 2.2 Hz, 1H), 7.81 (d, J = 8.6 Hz, 1H), 7.76 (dd, J = 8.6, 2.2 Hz, 1H), 7.39 (d, J = 8.6 Hz, 2H ), 7.37–7.29 (m, 2H), 7.13 (d, J = 8.1 Hz, 2H), 7.05 (d, J = 1.6 Hz, 1H), 5.29 (dd, J = 9.7, 5.7 Hz, 1H), 3.94 (d,J=9.4Hz,2H),3.77(d,J=13.2Hz,2H), 3.26–3.12(m,2H),3.08–2.80(m,3H), 2.71–2.63(m,1H), 1.75 (dd, J=77.7, 8.9 Hz, 3H), 1.40-1.25 (m, 2H).
实施例56Example 56
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S，3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000275
Figure PCTCN2020130361-appb-000275
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S,3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2 -Di-tert-butyl dicarboxylate
Figure PCTCN2020130361-appb-000276
Figure PCTCN2020130361-appb-000276
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.17毫摩尔)和(2S，3S)-2-氨基-3-甲基戊-1-醇(24毫克，0.20毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(65毫克，0.5毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.17 mmol) and (2S, 3S)-2-amino-3-methylpentan-1-ol (24 mg, 0.20 mmol) in tetrahydrofuran (5.0 mL) was added dropwise N,N-diisopropylethylamine (65 mg, 0.5 mmol), after dripping, react at 65°C overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/3)。得到71毫克浅黄色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S，3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：46.4％)。LCMS:RT＝4.29min,[M+H] +＝913.33。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/3). Obtain 71 mg of light yellow solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1 -Yl)-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-1 , Di-tert-butyl 2-dicarboxylic acid (yield: 46.4%). LCMS: RT = 4.29 min, [M+H] + =913.33.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S,3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxy acid
Figure PCTCN2020130361-appb-000277
Figure PCTCN2020130361-appb-000277
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S，3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(71毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2 -Di-tert-butyl dicarboxylate (71 mg, 0.07 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150 mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到76.82毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((2S，3S)-1-羟基-3-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：48.2％)。LCMS:RT＝3.40min,[M+H] +＝757.35。 1H NMR(500MHz,DMSO)δ12.94(s,1H),11.72(s,1H),10.14(s,1H),9.80(s,1H),8.44(s,1H),8.11–7.90(m,2H),7.90–7.70(m,2H),7.35(dd,J＝22.4,8.5Hz,4H),7.27–6.98(m,3H),6.01(d,J＝8.0Hz,1H),5.27(dd,J＝9.9,5.5Hz,1H),4.64(s,1H),3.74(s,2H),3.46(dd,J＝27.6,16.8Hz,3H),3.20(d,J＝11.1Hz,1H),3.05–2.93(m,1H),1.63–1.42(m,2H),1.23(s,1H),1.14–0.98(m,1H),0.85(t,J＝6.6Hz,5H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150 mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile from 5% liters To 100%; detection wavelength: 254nm. After purification, it was dried to obtain 76.82 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-(3-((2S,3S)-1-hydroxy-3-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indyl Dole-2-carboxylic acid (yield: 48.2%). LCMS: RT = 3.40 min, [M+H] + =757.35. 1 H NMR (500MHz, DMSO) δ 12.94 (s, 1H), 11.72 (s, 1H), 10.14 (s, 1H), 9.80 (s, 1H), 8.44 (s, 1H), 8.11-7.90 (m ,2H),7.90–7.70(m,2H),7.35(dd,J=22.4,8.5Hz,4H),7.27–6.98(m,3H),6.01(d,J=8.0Hz,1H), 5.27( dd,J=9.9,5.5Hz,1H), 4.64(s,1H), 3.74(s,2H), 3.46(dd,J=27.6,16.8Hz,3H), 3.20(d,J=11.1Hz,1H ), 3.05–2.93 (m, 1H), 1.63–1.42 (m, 2H), 1.23 (s, 1H), 1.14–0.98 (m, 1H), 0.85 (t, J=6.6 Hz, 5H).
实施例57Example 57
合成(S)-5-(3-(4-(2-氧杂-7-氮杂螺[3.5]壬烷-7-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(3-(4-(2-oxa-7-azaspiro[3.5]nonane-7-carboxamido)phenyl)-2-(4-(5-chloro- 2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000278
Figure PCTCN2020130361-appb-000278
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(3-(4-(2-氧杂-7-氮杂螺[3.5]壬烷-7-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(3-(4-(2-oxa-7-azaspiro[3.5]nonane-7-carboxamido)phenyl)-2-(4-(5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester
Figure PCTCN2020130361-appb-000279
Figure PCTCN2020130361-appb-000279
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(90毫克，0.10毫摩尔)和2-氧杂-7-氮杂螺[3.5]壬烷(38毫克，0.30毫摩尔)的四氢呋喃(2.0毫升)中滴加三乙胺(30毫克，0.30毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (90 mg, 0.10 mmol) and Triethylamine (30 mg, 0.30 mmol) was added dropwise to 2-oxa-7-azaspiro[3.5]nonane (38 mg, 0.30 mmol) in tetrahydrofuran (2.0 mL), and the reaction was completed at 65 degrees Celsius. overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/5)。得到90毫克浅黄色固体(S)-5-(3-(4-(2-氧杂-7-氮杂螺[3.5]壬烷-7-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：96.7％)。LCMS:RT＝4.24min,[M+H] +＝923.28。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/5). Obtain 90 mg of light yellow solid (S)-5-(3-(4-(2-oxa-7-azaspiro[3.5]nonane-7-carboxamido)phenyl)-2-(4- (5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate (yield: 96.7%). LCMS: RT = 4.24 min, [M+H] + =923.28.
步骤B：合成(S)-5-(3-(4-(2-氧杂-7-氮杂螺[3.5]壬烷-7-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(3-(4-(2-oxa-7-azaspiro[3.5]nonane-7-carboxamido)phenyl)-2-(4-(5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000280
Figure PCTCN2020130361-appb-000280
室温下，将(S)-5-(3-(4-(2-氧杂-7-氮杂螺[3.5]壬烷-7-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(90毫克，0.09毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(3-(4-(2-oxa-7-azaspiro[3.5]nonane-7-carboxamido)phenyl)-2-(4-(5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester (90 mg, 0.09 mmol) was added to methylene chloride (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到24毫克白色固体(S)-5-(3-(4-(2-氧杂-7-氮杂螺[3.5]壬烷-7-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸(收率：48.2％)。LCMS:RT＝3.29min,[M+H] +＝767.29。 1H NMR(500MHz,DMSO)δ12.94(s,1H),11.73(s,1H),10.14(s,1H),9.80(d,J＝5.9Hz,1H),8.40(d,J＝35.0Hz,1H),8.00(s,1H),7.95(s,1H),7.87–7.66(m,2H),7.37(ddd,J＝26.1,8.8,3.9Hz,4H),7.23–7.11(m,2H),7.07(d,J＝12.5Hz,1H),5.32–5.25(m,1H),3.49(s,2H),3.47–3.33(m,4H),3.30(d,J＝9.1Hz,2H),3.22(d,J＝9.3Hz,2H),3.00(s,1H),1.96(d,J＝49.5Hz,1H),1.60–1.18(m,6H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 24 mg of white solid (S)-5-(3-(4-(2-oxa-7-azaspiro[3.5]nonane-7-carboxamido)phenyl)-2- (4-(5-Chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2- Carboxylic acid (yield: 48.2%). LCMS: RT = 3.29 min, [M+H] + =767.29. 1 H NMR (500MHz, DMSO) δ 12.94 (s, 1H), 11.73 (s, 1H), 10.14 (s, 1H), 9.80 (d, J = 5.9 Hz, 1H), 8.40 (d, J = 35.0 Hz,1H),8.00(s,1H),7.95(s,1H),7.87–7.66(m,2H),7.37(ddd,J=26.1,8.8,3.9Hz,4H),7.23–7.11(m, 2H), 7.07(d,J=12.5Hz,1H), 5.32–5.25(m,1H), 3.49(s,2H), 3.47–3.33(m,4H), 3.30(d,J=9.1Hz,2H ), 3.22 (d, J = 9.3 Hz, 2H), 3.00 (s, 1H), 1.96 (d, J = 49.5 Hz, 1H), 1.60-1.18 (m, 6H).
实施例58Example 58
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(吗啉-4-羰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000281
Figure PCTCN2020130361-appb-000281
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(吗啉-4-羰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000282
Figure PCTCN2020130361-appb-000282
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.12毫摩尔)和吗啉代(哌啶-4-基)甲酮(154毫克，0.49毫摩尔)的四氢呋喃(2.0毫升)中滴加N,N-二异丙基乙胺(92毫克，0.72毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.12 mmol) and Morpholino(piperidin-4-yl)methanone (154mg, 0.49mmol) in tetrahydrofuran (2.0ml) was added dropwise with N,N-diisopropylethylamine (92mg, 0.72mmol), dropwise After that, react at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/3)。得到100毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(吗啉-4-羰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：96.7％)。LCMS:RT＝4.19min,[M+H] +＝994.37。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/3). Obtain 100 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl ester (yield: 96.7%). LCMS: RT = 4.19 min, [M+H] + =994.37.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(吗啉-4-羰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000283
Figure PCTCN2020130361-appb-000283
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(吗啉-4-羰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(100毫克，0.1毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl The ester (100 mg, 0.1 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到20毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(吗啉-4-羰基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：23.8％)。LCMS:RT＝3.23min,[M+H] +＝838.21。 1H NMR(500MHz,DMSO)δ13.11(s,1H),11.51(s,1H),10.15(d,J＝29.0Hz,1H),9.78(s,1H),8.45(s,1H),8.09–7.90(m,2H),7.87–7.72(m,2H),7.50(t,J＝8.6Hz,1H),7.45–7.31(m,3H),7.29–7.01(m,3H),5.68(t,J＝7.8Hz,1H),5.39–5.17(m,2H),4.21–4.08(m,3H),3.89–3.69(m,4H),3.25–3.19(m,1H),3.07–2.99(m,1H),2.84(t,J＝11.7Hz,3H),2.25(d,J＝6.8Hz,1H),2.10(d,J＝20.3Hz,1H),1.99(ddd,J＝14.3,13.2,6.1Hz,2H),1.87–1.81(m,1H),1.63(d,J＝11.1Hz,2H)。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid is precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 20 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2 -Carboxylic acid (yield: 23.8%). LCMS: RT = 3.23 min, [M+H] + =838.21. 1 H NMR (500MHz, DMSO) δ 13.11 (s, 1H), 11.51 (s, 1H), 10.15 (d, J = 29.0 Hz, 1H), 9.78 (s, 1H), 8.45 (s, 1H), 8.09–7.90(m,2H),7.87–7.72(m,2H),7.50(t,J=8.6Hz,1H),7.45–7.31(m,3H),7.29–7.01(m,3H),5.68( t,J=7.8Hz,1H),5.39–5.17(m,2H),4.21–4.08(m,3H), 3.89–3.69(m,4H), 3.25–3.19(m,1H),3.07–2.99( m, 1H), 2.84 (t, J = 11.7 Hz, 3H), 2.25 (d, J = 6.8 Hz, 1H), 2.10 (d, J = 20.3 Hz, 1H), 1.99 (ddd, J = 14.3, 13.2 , 6.1Hz, 2H), 1.87-1.81 (m, 1H), 1.63 (d, J=11.1Hz, 2H).
实施例59Example 59
合成5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(苯基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3-(4- (3-(phenylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000284
Figure PCTCN2020130361-appb-000284
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(苯磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(Benzenesulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000285
Figure PCTCN2020130361-appb-000285
将(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.19毫摩尔)，苯磺酰异氰酸酯(39毫克，0.21毫摩尔)溶于二氯甲烷(5.0毫升)中，加入吡啶(61毫克，0.78毫摩尔)。室温反应18小时。Add (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-diox Piperazin-1-yl) propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.19 mmol), benzenesulfonyl isocyanate (39 mg, 0.21 mmol) Dissolve in dichloromethane (5.0 mL) and add pyridine (61 mg, 0.78 mmol). React at room temperature for 18 hours.
加水淬灭反应，加乙酸乙酯(20毫升)稀释，水和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏，所得残余物用TLC板纯化得100毫克浅黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(苯磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：54.0％)。LCMS:RT＝4.36min,[M-H] -＝951.18。 The reaction was quenched by adding water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml × 3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by TLC plate to obtain 100 mg of light yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-(3-(Benzenesulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 54.0%). LCMS: RT = 4.36 min, [MH] - =951.18.
步骤B：合成5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(苯基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-(3-(phenylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000286
Figure PCTCN2020130361-appb-000286
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(苯磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(100毫克，0.10毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-(phenylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (100 mg, 0.10 mmol) Add dichloromethane (2.0 ml), add trifluoroacetic acid (0.5 ml) dropwise, and react at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到2毫克白色固体5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-(苯基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：2.4％)。LCMS:RT＝3.51min,[M-H] -＝795.23。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, dry to obtain 2 mg of white solid 5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(3-(phenylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 2.4%). LCMS: RT = 3.51 min, [MH] - =795.23.
实施例60Example 60
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(吗啉)-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(morpholine)-4-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000287
Figure PCTCN2020130361-appb-000287
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(吗啉)-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(morpholine)-4-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000288
Figure PCTCN2020130361-appb-000288
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.17毫摩尔)和吗啉(22毫克，0.25毫摩尔)的四氢呋喃(5.0毫升)中滴加三乙胺(51毫克，0.50毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.17 mmol) and Triethylamine (51 mg, 0.50 mmol) was added dropwise to morpholine (22 mg, 0.25 mmol) in tetrahydrofuran (5.0 ml), the dripping was completed, and the reaction was carried out at 65°C overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到88毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(吗啉)-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：59.5％)。LCMS:RT＝3.77min,[M+H] +＝883.24。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 88 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(morpholine)-4-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (Yield: 59.5%) . LCMS: RT = 3.77 min, [M+H] + = 883.24.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(吗啉)-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(morpholine)-4-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000289
Figure PCTCN2020130361-appb-000289
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(吗啉)-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(88毫克，0.1毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(morpholine)-4-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (88 mg, 0.1 mmol) Add dichloromethane (2.0 mL), add trifluoroacetic acid (0.5 mL) dropwise, and react at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到26.9毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(吗啉)-4-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：37.4％)。LCMS:RT＝3.25min,[M+H] +＝727.17。 1H NMR(500MHz,DMSO)δ12.92(s,1H),11.72(s,1H),10.14(s,1H),9.77(d,J＝23.4Hz,1H),8.51(s,1H),8.00(s,1H),7.95(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.2Hz,1H),7.41(d,J＝8.5Hz,2H),7.37(d,J＝8.9Hz,1H),7.32(d,J＝7.3Hz,1H),7.17(t,J＝14.2Hz,2H),7.05(s,1H),5.30(dd,J＝9.8,5.8Hz,1H),3.74(s,2H),3.65–3.54(m,5H),3.44–3.39(m,4H),3.29–3.17(m,2H),3.06–2.98(m,1H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 26.9 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-(morpholine)-4-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 37.4%). LCMS: RT = 3.25 min, [M+H] + =727.17. 1 H NMR (500MHz, DMSO) δ 12.92 (s, 1H), 11.72 (s, 1H), 10.14 (s, 1H), 9.77 (d, J = 23.4 Hz, 1H), 8.51 (s, 1H), 8.00 (s, 1H), 7.95 (d, J = 2.2 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.77 (dd, J = 8.6, 2.2 Hz, 1H), 7.41 (d, J =8.5Hz, 2H), 7.37 (d, J = 8.9Hz, 1H), 7.32 (d, J = 7.3Hz, 1H), 7.17 (t, J = 14.2Hz, 2H), 7.05 (s, 1H), 5.30(dd,J=9.8,5.8Hz,1H), 3.74(s,2H), 3.65–3.54(m,5H), 3.44–3.39(m,4H), 3.29–3.17(m,2H),3.06– 2.98 (m, 1H).
实施例61Example 61
合成(S)-5-(3-(4-(4-氨基甲酰基-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(3-(4-(4-carbamoyl-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H-tetrazole-1- (Yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000290
Figure PCTCN2020130361-appb-000290
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(3-(4-(4-氨基甲酰基-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2，3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(3-(4-(4-carbamoyl-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H-tetrazole) -1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000291
Figure PCTCN2020130361-appb-000291
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.17毫摩尔)和哌啶-4-甲酰胺(32毫克，0.25毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(65毫克，0.50毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.17 mmol) and Piperidine-4-carboxamide (32 mg, 0.25 mmol) in tetrahydrofuran (5.0 ml) was added dropwise with N,N-diisopropylethylamine (65 mg, 0.50 mmol), after dripping, react at 65 degrees Celsius overnight .
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得 到68毫克黄色固体(S)-5-(3-(4-(4-氨基甲酰基-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：43.8％)。LCMS:RT＝4.10min,[M+H] +＝924.27。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 68 mg of yellow solid (S)-5-(3-(4-(4-carbamoyl-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H-tetra (Azol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (Yield: 43.8 %). LCMS: RT = 4.10 min, [M+H] + =924.27.
步骤B：合成(S)-5-(3-(4-(4-氨基甲酰基-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(3-(4-(4-carbamoyl-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H-tetrazole) -1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000292
Figure PCTCN2020130361-appb-000292
室温下，将(S)-5-(3-(4-(4-氨基甲酰基-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2，3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(68毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(3-(4-(4-carbamoyl-1-carboxamido)phenyl)-2-(4-(5-chloro-2-(1H-tetrazole) -1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (68 mg, 0.07 mg Mol) was added to dichloromethane (2.0 ml), trifluoroacetic acid (0.5 ml) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到56毫克白色固体(S)-5-(3-(4-(4-氨基甲酰基-1-甲酰氨基)苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-2-羧酸(收率：37.4％)。LCMS:RT＝3.09min,[M+H] +＝768.27。 1H NMR(500MHz,DMSO)δ12.93(s,1H),11.73(s,1H),10.14(s,1H),9.79(s,1H),8.45(s,1H),8.01(s,1H),7.95(d,J＝2.2Hz,1H),7.82(d,J＝8.6Hz,1H),7.77(dd,J＝8.6,2.2Hz,1H),7.36(ddd,J＝12.5,10.6,5.1Hz,4H),7.27(s,1H),7.20–7.11(m,2H),7.07(d,J＝9.5Hz,1H),6.79(s,1H),5.30(dd,J＝9.7,5.9Hz,1H),4.10(d,J＝13.0Hz,3H),3.74(s,1H),3.22(d,J＝9.4Hz,2H),3.05–2.98(m,1H),2.78(t,J＝11.8Hz,2H),2.29(t,J＝11.5Hz,1H),1.70(d,J＝12.0Hz,3H),1.46(d,J＝12.2Hz,3H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 56 mg of white solid (S)-5-(3-(4-(4-carbamoyl-1-carboxamido)phenyl)-2-(4-(5-chloro-2- (1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)propionamido)-1H-indole-2-carboxylic acid (yield: 37.4%). LCMS: RT = 3.09 min, [M+H] + =768.27. 1 H NMR (500MHz, DMSO) δ 12.93 (s, 1H), 11.73 (s, 1H), 10.14 (s, 1H), 9.79 (s, 1H), 8.45 (s, 1H), 8.01 (s, 1H) ), 7.95 (d, J = 2.2 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.77 (dd, J = 8.6, 2.2 Hz, 1H), 7.36 (ddd, J = 12.5, 10.6, 5.1Hz, 4H), 7.27 (s, 1H), 7.20-7.11 (m, 2H), 7.07 (d, J = 9.5 Hz, 1H), 6.79 (s, 1H), 5.30 (dd, J = 9.7, 5.9 Hz, 1H), 4.10 (d, J = 13.0 Hz, 3H), 3.74 (s, 1H), 3.22 (d, J = 9.4 Hz, 2H), 3.05-2.98 (m, 1H), 2.78 (t, J = 11.8 Hz, 2H), 2.29 (t, J = 11.5 Hz, 1H), 1.70 (d, J = 12.0 Hz, 3H), 1.46 (d, J = 12.2 Hz, 3H).
实施例62Example 62
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲氧基甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(Methoxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000293
Figure PCTCN2020130361-appb-000293
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲氧基甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(Methoxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000294
Figure PCTCN2020130361-appb-000294
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.17毫摩尔)和4-(甲氧基甲基)哌啶(66毫克，0.51毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(65毫克，0.50毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.17 mmol) and 4-(Methoxymethyl)piperidine (66 mg, 0.51 mmol) in tetrahydrofuran (5.0 mL) was added dropwise with N,N-diisopropylethylamine (65 mg, 0.50 mmol). React at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/4)。得到90毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲氧基甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：57.7％)。LCMS:RT＝3.92min,[M+H] +＝925.25。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/4). Obtain 90 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(4-(methoxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl Ester (yield: 57.7%). LCMS: RT = 3.92 min, [M+H] + =925.25.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲氧基甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(Methoxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000295
Figure PCTCN2020130361-appb-000295
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲氧基甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(90毫克，0.09毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(Methoxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (90 mg, 0.09 mmol) was added to methylene chloride (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到12毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(甲氧基甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：15.9％)。LCMS:RT＝3.44min,[M+H] +＝769.17。 1H NMR(400MHz,DMSO)δ12.90(s,1H),11.71(s,1H),10.13(s,1H),9.78(s,1H),8.40(s,1H),8.03–7.89(m,2H),7.77(dt,J＝8.6,5.4Hz,2H),7.40(d,J＝8.5Hz,2H),7.37–7.28(m,2H),7.13(d,J＝8.2Hz,2H),7.05(d,J＝1.5Hz,1H),5.29(dd,J＝9.7,5.8Hz,1H),4.10(d,J＝13.6Hz,2H),3.74(s,2H),3.26–3.16(m,6H),3.04–2.96(m,1H),2.72(dd,J＝27.3,15.6Hz,3H),2.06–1.93(m,1H),1.81–1.58(m,4H),1.45(s,1H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 12 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-(4-(Methoxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid Rate: 15.9%). LCMS: RT = 3.44 min, [M+H] + =769.17. 1 H NMR (400MHz, DMSO) δ 12.90 (s, 1H), 11.71 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.40 (s, 1H), 8.03 - 7.89 (m , 2H), 7.77 (dt, J = 8.6, 5.4 Hz, 2H), 7.40 (d, J = 8.5 Hz, 2H), 7.37-7.28 (m, 2H), 7.13 (d, J = 8.2 Hz, 2H) ,7.05(d,J=1.5Hz,1H), 5.29(dd,J=9.7,5.8Hz,1H), 4.10(d,J=13.6Hz,2H), 3.74(s,2H), 3.26–3.16( m,6H),3.04–2.96(m,1H),2.72(dd,J=27.3,15.6Hz,3H),2.06–1.93(m,1H),1.81–1.58(m,4H),1.45(s, 1H).
实施例63Example 63
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000296
Figure PCTCN2020130361-appb-000296
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(Hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000297
Figure PCTCN2020130361-appb-000297
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨 基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.17毫摩尔)和4-羟甲基哌啶(65毫克，0.51毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(65毫克，0.50毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.17 mmol) and 4-Hydroxymethylpiperidine (65 mg, 0.51 mmol) in tetrahydrofuran (5.0 mL) was added dropwise with N,N-diisopropylethylamine (65 mg, 0.50 mmol), after dripping, react at 65 degrees Celsius overnight .
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到75毫克黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：49.0％)。LCMS:RT＝4.29min,[M+H] +＝911.35。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/1). Obtain 75 mg of yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-(4-(hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester ( Yield: 49.0%). LCMS: RT = 4.29 min, [M+H] + = 911.35.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000298
Figure PCTCN2020130361-appb-000298
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(75.0毫克，0.08毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(Hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (75.0 Mg, 0.08 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到8.5毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(羟甲基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：13.1％)。LCMS:RT＝3.20min,[M+H] +＝755.31。 1H NMR(400MHz,DMSO)δ12.91(s,1H),11.71(s,1H),10.13(s,1H),9.78(s,1H),8.40(s,1H),8.00(s,1H),7.94(d,J＝2.2Hz,1H),7.77(dt,J＝8.6,5.4Hz,2H),7.47–7.24(m,4H),7.20–7.02(m,3H),6.97(s,1H),5.29(dd,J＝9.7,5.8Hz,1H),4.11(d,J＝13.0Hz,2H),3.92(s,2H),3.73(s,2H),3.25(d,J＝6.2Hz,2H),3.13–2.85(m,2H),2.71(dd,J＝24.1,13.0Hz,3H),1.65(d,J＝12.9Hz,2H),1.55(s,1H),1.05(dd,J＝21.3,11.9Hz,2H)。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 8.5 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3- Dioxopiperazin-1-yl)-3-(4-(4-(hydroxymethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (Yield: 13.1%). LCMS: RT = 3.20 min, [M+H] + =755.31. 1 H NMR (400MHz, DMSO) δ 12.91 (s, 1H), 11.71 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.40 (s, 1H), 8.00 (s, 1H) ),7.94(d,J=2.2Hz,1H),7.77(dt,J=8.6,5.4Hz,2H),7.47–7.24(m,4H),7.20–7.02(m,3H),6.97(s, 1H), 5.29 (dd, J = 9.7, 5.8 Hz, 1H), 4.11 (d, J = 13.0 Hz, 2H), 3.92 (s, 2H), 3.73 (s, 2H), 3.25 (d, J = 6.2 Hz, 2H), 3.13–2.85 (m, 2H), 2.71 (dd, J = 24.1, 13.0 Hz, 3H), 1.65 (d, J = 12.9 Hz, 2H), 1.55 (s, 1H), 1.05 (dd , J=21.3, 11.9 Hz, 2H).
实施例64Example 64
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((S))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000299
Figure PCTCN2020130361-appb-000299
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester
Figure PCTCN2020130361-appb-000300
Figure PCTCN2020130361-appb-000300
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.17毫摩尔)和(S)-吡咯烷-3-基甲醇(34毫克，0.34毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(65毫克，0.50毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.17 mmol) and (S)-Pyrrolidin-3-ylmethanol (34 mg, 0.34 mmol) in tetrahydrofuran (5.0 mL) was added dropwise with N,N-diisopropylethylamine (65 mg, 0.50 mmol). React at 65°C overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/3)。得到85毫克黄色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：56.3％)。LCMS:RT＝4.29min,[M+H] +＝897.42。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/3). Obtain 85 mg of yellow solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((S))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester (yield: 56.3%). LCMS: RT = 4.29 min, [M+H] + =897.42.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000301
Figure PCTCN2020130361-appb-000301
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(85.0毫克，0.08毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((S))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (85.0 mg, 0.08 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到5.7毫克白色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((S))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：13.1％)。LCMS:RT＝3.14min,[M-H] -＝741.25。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, dry to obtain 5.7 mg of white solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-((S))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxy Acid (yield: 13.1%). LCMS: RT = 3.14 min, [MH] - = 741.25.
实施例65Example 65
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((R))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000302
Figure PCTCN2020130361-appb-000302
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester
Figure PCTCN2020130361-appb-000303
Figure PCTCN2020130361-appb-000303
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(180毫克，0.20毫摩尔)和(R)-吡咯烷-3-基甲醇(24毫克，0.24毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(65毫克，0.50毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (180 mg, 0.20 mmol) and (R)-Pyrrolidin-3-ylmethanol (24 mg, 0.24 mmol) in tetrahydrofuran (5.0 mL) was added dropwise with N,N-diisopropylethylamine (65 mg, 0.50 mmol). React at 65°C overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/3)。得到90毫克黄色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：49.7％)。LCMS:RT＝4.19min,[M+H] +＝897.33。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/3). Obtain 90 mg of yellow solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((R))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester (yield: 49.7%). LCMS: RT = 4.19 min, [M+H] + =897.33.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000304
Figure PCTCN2020130361-appb-000304
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(90毫克，0.08毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((R))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (90 mg, 0.08 mmol) was added to methylene chloride (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到56毫克浅黄色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((R))-3-(羟甲基)吡咯烷-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：75.1％)。LCMS:RT＝3.16min,[M+H] +＝741.17。 1H NMR(400MHz,DMSO)δ12.93(s,1H),11.71(s,1H),10.13(s,1H),9.78(s,1H),8.05(s,1H),7.99(s, 1H),7.94(d,J＝2.2Hz,1H),7.81(d,J＝8.6Hz,1H),7.76(dd,J＝8.6,2.2Hz,1H),7.45(d,J＝8.5Hz,2H),7.33(dt,J＝8.9,5.4Hz,2H),7.13(d,J＝7.9Hz,2H),7.05(d,J＝1.6Hz,1H),5.29(dd,J＝9.7,5.8Hz,1H),3.95(s,2H),3.73(s,2H),3.42(dd,J＝7.0,3.5Hz,5H),3.26–3.09(m,3H),3.05–2.94(m,1H),2.31(dt,J＝14.1,7.1Hz,1H),1.90(td,J＝12.1,7.2Hz,1H),1.62(td,J＝15.4,7.7Hz,1H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 56 mg of light yellow solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxo Piperazin-1-yl)-3-(4-((R))-3-(hydroxymethyl)pyrrolidine-1-carboxamido)phenyl)propionamido)-1H-indole-2- Carboxylic acid (yield: 75.1%). LCMS: RT = 3.16 min, [M+H] + =741.17. 1 H NMR (400MHz, DMSO) δ 12.93 (s, 1H), 11.71 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.05 (s, 1H), 7.99 (s, 1H) ), 7.94 (d, J = 2.2 Hz, 1H), 7.81 (d, J = 8.6 Hz, 1H), 7.76 (dd, J = 8.6, 2.2 Hz, 1H), 7.45 (d, J = 8.5 Hz, 2H ), 7.33 (dt, J = 8.9, 5.4 Hz, 2H), 7.13 (d, J = 7.9 Hz, 2H), 7.05 (d, J = 1.6 Hz, 1H), 5.29 (dd, J = 9.7, 5.8 Hz ,1H),3.95(s,2H),3.73(s,2H),3.42(dd,J=7.0,3.5Hz,5H), 3.26–3.09(m,3H),3.05–2.94(m,1H), 2.31 (dt, J=14.1, 7.1 Hz, 1H), 1.90 (td, J=12.1, 7.2 Hz, 1H), 1.62 (td, J=15.4, 7.7 Hz, 1H).
实施例66Example 66
合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-4-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(3-((S)-1-hydroxy-4-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000305
Figure PCTCN2020130361-appb-000305
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-4-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-1-hydroxy-4-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
Figure PCTCN2020130361-appb-000306
Figure PCTCN2020130361-appb-000306
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(235毫克，0.26毫摩尔)和(S)-2-氨基-4-甲基戊-1-醇(62毫克，0.53毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(101毫克，0.78毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (235 mg, 0.26 mmol) and (S)-2-Amino-4-methylpentan-1-ol (62 mg, 0.53 mmol) in tetrahydrofuran (5.0 mL) was added dropwise N,N-diisopropylethylamine (101 mg, 0.78 mmol) Mol), after dripping, react overnight at 65 degrees Celsius.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/3)。得到68毫克黄色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-4-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：28.2％)。LCMS:RT＝4.29min,[M+H] +＝913.43。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/3). Obtain 68 mg of yellow solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(3-((S)-1-hydroxy-4-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2- Di-tert-butyl dicarboxylate (yield: 28.2%). LCMS: RT = 4.29 min, [M+H] + =913.43.
步骤B：合成5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-4-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-1-hydroxy-4-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000307
Figure PCTCN2020130361-appb-000307
室温下，将5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-4-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(68毫克，0.07毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, the 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(3-((S)-1-hydroxy-4-methylpent-2-yl)ureido)phenyl)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate (68 mg, 0.07 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用溶于二氯甲烷(1.0毫升)中，将其滴加入正己烷(10.0毫升)中，析出白色固体，抽滤，滤饼用正己烷洗涤，干燥得到19.44毫克浅红色固体5-((S)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(3-((S)-1-羟基-4-甲基戊-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：75.1％)。LCMS:RT＝3.40min,[M-H] -＝757.17。 1H NMR(400MHz,DMSO)δ12.89(s,1H),11.71(s,1H),10.12(s,1H), 9.78(s,1H),8.38(s,1H),8.03–7.90(m,2H),7.77(dt,J＝8.6,5.4Hz,2H),7.35(dd,J＝19.7,8.6Hz,4H),7.22–7.01(m,3H),5.93(d,J＝8.4Hz,1H),5.27(dd,J＝9.9,5.5Hz,1H),3.88(s,1H),3.73–3.51(m,5H),3.47–3.25(m,3H),3.14(ddd,J＝17.1,14.8,8.5Hz,2H),3.06–2.93(m,1H),1.63(dt,J＝13.3,6.5Hz,1H),1.36–1.04(m,6H),0.87(t,J＝6.9Hz,6H)。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was dissolved in dichloromethane (1.0 ml) and added dropwise to n-hexane (10.0 ml). A white solid precipitated out and filtered with suction. The filter cake was washed with n-hexane and dried to obtain 19.44 mg of light red solid 5-((S)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3 -Dioxopiperazin-1-yl)-3-(4-(3-((S)-1-hydroxy-4-methylpent-2-yl)ureido)phenyl)propionamido)- 1H-Indole-2-carboxylic acid (yield: 75.1%). LCMS: RT = 3.40 min, [MH] - = 757.17. 1 H NMR (400MHz, DMSO) δ 12.89 (s, 1H), 11.71 (s, 1H), 10.12 (s, 1H), 9.78 (s, 1H), 8.38 (s, 1H), 8.03-7.90 (m , 2H), 7.77 (dt, J = 8.6, 5.4 Hz, 2H), 7.35 (dd, J = 19.7, 8.6 Hz, 4H), 7.22-7.01 (m, 3H), 5.93 (d, J = 8.4 Hz, 1H), 5.27(dd,J=9.9,5.5Hz,1H), 3.88(s,1H), 3.73–3.51(m,5H), 3.47–3.25(m,3H), 3.14(ddd,J=17.1, 14.8,8.5Hz,2H),3.06–2.93(m,1H),1.63(dt,J=13.3,6.5Hz,1H),1.36–1.04(m,6H),0.87(t,J=6.9Hz,6H ).
实施例67Example 67
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-羟乙基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-(4-(2-hydroxyethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000308
Figure PCTCN2020130361-appb-000308
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-羟乙基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-Hydroxyethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000309
Figure PCTCN2020130361-appb-000309
室温下，向含有(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((苯氧基羰基)氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.16毫摩尔)和2-(哌啶-4-基)乙-1-醇(65毫克，0.50毫摩尔)的四氢呋喃(5.0毫升)中滴加N,N-二异丙基乙胺(50毫克，0.50毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionylamino)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.16 mmol) and 2-(piperidin-4-yl)ethan-1-ol (65 mg, 0.50 mmol) in tetrahydrofuran (5.0 ml) was added dropwise with N,N-diisopropylethylamine (50 mg, 0.50 mmol) , After dripping, react at 65 degrees Celsius overnight.
反应结束，加水淬灭，混合液用乙酸乙酯(10毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/2)。得到75毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-羟乙基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：48.1％)。LCMS:RT＝4.29min,[M+H] +＝925.45。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (10 ml×3 times), and the organic phases were combined. The organic phase was washed with saturated brine (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 75 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazine-1- Yl)-3-(4-(4-(2-hydroxyethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl Esters (yield: 48.1%). LCMS: RT = 4.29 min, [M+H] + =925.45.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-羟乙基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-hydroxyethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000310
Figure PCTCN2020130361-appb-000310
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-羟乙基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(75.0毫克，0.08毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-(4-(2-Hydroxyethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (75.0 mg, 0.08 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到8.5毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-(4-(2-羟乙基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：75.1％)。LCMS:RT＝3.23min,[M+H] +＝ 769.13。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, dry to obtain 8.5 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-(4-(2-hydroxyethyl)piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid Rate: 75.1%). LCMS: RT = 3.23 min, [M+H] + = 769.13.
实施例68Example 68
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N-乙基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((N-ethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000311
Figure PCTCN2020130361-appb-000311
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N-乙基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((N-ethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000312
Figure PCTCN2020130361-appb-000312
将(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克，0.23毫摩尔)，甲基氨磺酰氯(32毫克，0.23毫摩尔)溶于二氯甲烷(5.0毫升)中，加入N,N-二异丙基乙胺(58毫克，0.45毫摩尔)。室温反应18小时。Add (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-diox Piperazin-1-yl) propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.23 mmol), methyl sulfonyl chloride (32 mg, 0.23 mmol) ) Was dissolved in dichloromethane (5.0 mL), and N,N-diisopropylethylamine (58 mg, 0.45 mmol) was added. React at room temperature for 18 hours.
加水淬灭反应，加乙酸乙酯(20毫升)稀释，水和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏，所得残余物用TLC板纯化得20毫克浅黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N-乙基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：10.1％)。LCMS:RT＝4.29min,[M-H] -＝875.21。 The reaction was quenched with water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml × 3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by TLC plate to obtain 20 mg of light yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-((N-ethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 10.1%). LCMS: RT = 4.29 min, [MH] - = 875.21.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N-乙基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((N-ethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000313
Figure PCTCN2020130361-appb-000313
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N-乙基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(20.0毫克，0.02毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((N-ethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (20.0 mg, 0.02 mmol ) Was added to dichloromethane (2.0 ml), trifluoroacetic acid (0.5 ml) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到15.5毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N-乙基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：94.3％)。LCMS:RT＝3.39min,[M+H] +＝721.12。 1H NMR(400MHz,DMSO)δ12.91(s,1H),11.71(s,1H),10.12(s,1H),9.78(s,1H),9.57(s,1H),8.01–7.89(m,2H),7.78(dt,J＝8.6,5.4Hz,2H),7.41–7.26(m,3H),7.15(dd,J＝30.7,8.5Hz,4H),7.06(t,J＝3.7Hz,1H),5.30(dd,J＝9.6,6.0Hz, 1H),3.72(s,4H),3.26–3.17(m,1H),3.07–2.97(m,1H),2.89–2.78(m,2H),0.96–0.80(m,3H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 15.5 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper Azin-1-yl)-3-(4-((N-ethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 94.3%). LCMS: RT = 3.39 min, [M+H] + = 721.12. 1 H NMR (400MHz, DMSO) δ 12.91 (s, 1H), 11.71 (s, 1H), 10.12 (s, 1H), 9.78 (s, 1H), 9.57 (s, 1H), 8.01-7.89 (m ,2H),7.78(dt,J=8.6,5.4Hz,2H),7.41-7.26(m,3H),7.15(dd,J=30.7,8.5Hz,4H),7.06(t,J=3.7Hz, 1H), 5.30(dd,J=9.6,6.0Hz, 1H), 3.72(s,4H), 3.26–3.17(m,1H),3.07–2.97(m,1H), 2.89–2.78(m,2H) ,0.96–0.80(m,3H).
实施例69Example 69
合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N,N-二甲基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3 -(4-((N,N-Dimethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000314
Figure PCTCN2020130361-appb-000314
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N,N-二甲基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁基酯Step A: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((N,N-Dimethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020130361-appb-000315
Figure PCTCN2020130361-appb-000315
将(S)-5-(3-(4-氨基苯基)-2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(180毫克，0.23毫摩尔)，甲基氨磺酰氯(40毫克，0.28毫摩尔)溶于二氯甲烷(5.0毫升)中，加入吡啶(37毫克，0.46毫摩尔)。室温反应2小时。Add (S)-5-(3-(4-aminophenyl)-2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-diox Piperazin-1-yl) propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (180 mg, 0.23 mmol), methyl sulfonyl chloride (40 mg, 0.28 mmol) ) Was dissolved in dichloromethane (5.0 mL), and pyridine (37 mg, 0.46 mmol) was added. React at room temperature for 2 hours.
加水淬灭反应，加乙酸乙酯(20毫升)稀释，水和食盐水(10毫升×3次)洗涤，无水硫酸钠干燥，减压蒸馏，所得残余物用TLC板纯化得85毫克浅黄色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N,N-二甲基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁基酯(收率41.5％)。LCMS:RT＝4.32min,[M-H] -＝875.13。 The reaction was quenched with water, diluted with ethyl acetate (20 ml), washed with water and brine (10 ml × 3 times), dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by TLC plate to obtain 85 mg of light yellow solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl)-3- (4-((N,N-Dimethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield 41.5%). LCMS: RT = 4.32 min, [MH] - = 875.13.
步骤B：合成(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N,N-二甲基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((N,N-dimethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020130361-appb-000316
Figure PCTCN2020130361-appb-000316
室温下，将(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N,N-二甲基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁基酯(85毫克，0.09毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(0.5毫升)，室温反应3小时。At room temperature, (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(4-((N,N-dimethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (85 mg , 0.09 mmol) was added to dichloromethane (2.0 mL), trifluoroacetic acid (0.5 mL) was added dropwise, and the reaction was carried out at room temperature for 3 hours.
将反应液减压蒸馏。将所得残余物用制备型高效液相色谱纯化。分离条件如下，色谱柱：X select C18 19mm*150mm；流动相：水(含有0.05％的三氟乙酸)和乙腈；流速：25毫升/分钟；梯度：在7分钟内，乙腈从5％升到100％；检测波长：254nm。纯化后，干燥得到24.3毫克白色固体(S)-5-(2-(4-(5-氯-2-(1H-四唑-1-基)苯基)-2,3-二氧代哌嗪-1-基)-3-(4-((N,N-二甲基氨磺酰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：34.3％)。LCMS:RT＝3.41min,[M-H] -＝721.31。 1H NMR(500MHz,DMSO)δ12.93(s,1H),11.73(s,1H),10.12(s,1H),9.80(d,J＝4.7Hz,2H),7.99(s,1H),7.94 (d,J＝2.2Hz,1H),7.79(dt,J＝8.6,5.4Hz,2H),7.37(d,J＝8.9Hz,1H),7.31(dd,J＝8.9,1.8Hz,1H),7.18(dd,J＝20.2,8.0Hz,4H),7.06(t,J＝5.2Hz,1H),5.34–5.26(m,1H),4.19(s,1H),3.73(s,2H),3.23(dd,J＝14.4,5.0Hz,1H),3.03(dd,J＝14.0,10.2Hz,1H),2.64(s,6H),1.99(dt,J＝12.5,6.9Hz,1H)。 The reaction liquid was distilled under reduced pressure. The obtained residue was purified by preparative high performance liquid chromatography. The separation conditions are as follows, chromatographic column: X select C18 19mm*150mm; mobile phase: water (containing 0.05% trifluoroacetic acid) and acetonitrile; flow rate: 25 ml/min; gradient: within 7 minutes, acetonitrile rises from 5% to 100%; detection wavelength: 254nm. After purification, it was dried to obtain 24.3 mg of white solid (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiper (Azin-1-yl)-3-(4-((N,N-dimethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 34.3% ). LCMS: RT = 3.41 min, [MH] - = 721.31. 1 H NMR (500MHz, DMSO) δ 12.93 (s, 1H), 11.73 (s, 1H), 10.12 (s, 1H), 9.80 (d, J = 4.7 Hz, 2H), 7.99 (s, 1H), 7.94 (d,J=2.2Hz,1H),7.79(dt,J=8.6,5.4Hz,2H),7.37(d,J=8.9Hz,1H),7.31(dd,J=8.9,1.8Hz,1H ), 7.18(dd,J=20.2,8.0Hz,4H),7.06(t,J=5.2Hz,1H),5.34-5.26(m,1H),4.19(s,1H),3.73(s,2H) , 3.23 (dd, J=14.4, 5.0 Hz, 1H), 3.03 (dd, J=14.0, 10.2 Hz, 1H), 2.64 (s, 6H), 1.99 (dt, J=12.5, 6.9 Hz, 1H).
实施例70：吸收光法检测本发明化合物对人凝血因子XIa抑制的生物活性Example 70: Detection of the biological activity of the compound of the present invention on the inhibition of human coagulation factor XIa by light absorption method
1、实验材料1. Experimental materials
酶：Human Factor XIa(ENZYME RESEARCH，货号HFXIa 1111a)Enzyme: Human Factor XIa (ENZYME Research, Item No. HFXIa 1111a)
底物：S-2366 TM：(CHROMOGENIX，货号82109039) Substrate: S-2366 TM : (CHROMOGENIX, article number 82109039)
缓冲液：145mM NaCl，5mM KCl，1mg/mL PEG 8000,，30mM HEPES，PH7.4Buffer: 145mM NaCl, 5mM KCl, 1mg/mL PEG 8000, 30mM HEPES, PH7.4
2、实验步骤2. Experimental steps
将溶于100％DMSO的10mM受试化合物用100％DMSO稀释至1000、200、40、8、1.6、0.32、0.064、0.0128、0.00256、0.00128μM；在96孔板中每孔加入98μL(77.7ng/mL)的FXIa酶溶液，空白孔加入98μL缓冲液代替，再加入2μL不同浓度的化合物，空白和对照孔用DMSO代替，用振荡器混匀，37℃孵育20min。The 10mM test compound dissolved in 100% DMSO was diluted with 100% DMSO to 1000, 200, 40, 8, 1.6, 0.32, 0.064, 0.0128, 0.00256, 0.00128μM; 98μL (77.7ng /mL) FXIa enzyme solution, the blank wells were replaced by 98μL of buffer, and then 2μL of compounds of different concentrations were added. The blank and control wells were replaced with DMSO, mixed with a shaker, and incubated at 37°C for 20min.
最后每孔加入800μM的底物100μL，在405nm处测其吸光度。Finally, 100μL of 800μM substrate was added to each well, and the absorbance was measured at 405nm.
3、数据处理3. Data processing
用GraphPad Prism软件进行曲线拟合，计算IC 50值，见表一。 Use GraphPad Prism software to perform curve fitting and calculate IC 50 value, see Table 1.
表一：本发明化合物对人FXIa抑制的IC50Table 1: The IC50 of the compound of the present invention for inhibiting human FXIa
Figure PCTCN2020130361-appb-000317
Figure PCTCN2020130361-appb-000317
Figure PCTCN2020130361-appb-000318
Figure PCTCN2020130361-appb-000318
结论：本发明化合物对人FXIa具有明显的抑制活性。Conclusion: The compound of the present invention has obvious inhibitory activity on human FXIa.
实施例71：:本发明化合物对人血浆体外抗凝血作用的测定Example 71: Determination of the anticoagulant effect of the compound of the present invention on human plasma in vitro
1、实验材料1. Experimental materials
血浆：人血收集于含3.2％柠檬酸钠(体积比1:9)的真空采血管中，室温3000rpm离心10min，收集血浆，分装在EP管中，-80℃保存。Plasma: Human blood is collected in a vacuum blood collection tube containing 3.2% sodium citrate (volume ratio 1:9), centrifuged at 3000 rpm for 10 minutes at room temperature, and the plasma is collected, aliquoted in EP tubes, and stored at -80°C.
试剂：APTT测定试剂盒(活化部分凝血活酶时间检测定剂盒，mindray)、氯化钙溶液。Reagents: APTT determination kit (activated partial thromboplastin time detection kit, mindray), calcium chloride solution.
仪器：凝血仪(mindray，C2000-A)Instrument: coagulometer (mindray, C2000-A)
2、实验方法2. Experimental method
取分装的冻存人血浆室温融化后，混合均匀。将溶于100％DMSO的10mM受试化合物用100％DMSO稀释至1500、750、375、187.5、93.75、46.88、23.44、11.72μM；在1.5mL EP管中加入98μL人血浆，再加入2μL不同浓度的化合物，空白组加入2μL 100％DMSO，37℃水浴孵育10min，将样品放入凝血仪中对应的位置，进行化合物的APTT测定。Take aliquots of cryopreserved human plasma after thawing at room temperature, and mix them evenly. Dilute the 10mM test compound dissolved in 100% DMSO with 100% DMSO to 1500, 750, 375, 187.5, 93.75, 46.88, 23.44, 11.72μM; add 98μL of human plasma to a 1.5mL EP tube, and then add 2μL of different concentrations Add 2 μL 100% DMSO to the blank group, and incubate in a 37°C water bath for 10 minutes. Put the sample into the corresponding position in the coagulometer to perform the APTT determination of the compound.
3、数据处理3. Data processing
用GraphPad Prism软件进行曲线拟合，分别计算EC1.5×和EC2×值，即1.5倍和2倍空白对照组的APTT所对应的化合物的浓度，结果见表二。GraphPad Prism software was used to perform curve fitting, and EC1.5× and EC2× values were calculated respectively, that is, 1.5 times and 2 times the concentration of the compound corresponding to the APTT of the blank control group. The results are shown in Table 2.
表二：本发明化合物对人血浆体外抗凝血作用Table 2: In vitro anticoagulant effects of the compounds of the present invention on human plasma
Figure PCTCN2020130361-appb-000319
Figure PCTCN2020130361-appb-000319
Figure PCTCN2020130361-appb-000320
Figure PCTCN2020130361-appb-000320
结论：从表二中可以看出本发明化合物对人血浆具有明显的抗凝血作用。Conclusion: It can be seen from Table 2 that the compound of the present invention has a significant anticoagulant effect on human plasma.
实施例72：本发明化合物的大鼠药代动力学特征考察Example 72: Investigation of rat pharmacokinetic characteristics of the compound of the present invention
1、实验材料1. Experimental materials
SD大鼠：雄性，180-250g，购于广东省医学实验动物中心。食蟹猴：雄性，4-6kg，购于广州春盛生物研究院有限公司。SD rats: male, 180-250g, purchased from Guangdong Medical Experimental Animal Center. Cynomolgus monkey: male, 4-6kg, purchased from Guangzhou Chunsheng Biological Research Institute Co., Ltd.
试剂：DMSO(二甲亚砜)，PEG-400(聚乙二醇400)，生理盐水，肝素，乙腈，甲酸，普萘洛尔(内标)均为市售可得。Reagents: DMSO (dimethyl sulfoxide), PEG-400 (polyethylene glycol 400), physiological saline, heparin, acetonitrile, formic acid, propranolol (internal standard) are all commercially available.
仪器：赛默飞LC-MS(U300 UPLC，TSQ QUANTUMN ULTRA三重四级杆质谱)。Instrument: Thermo Fisher LC-MS (U300 UPLC, TSQ QUANTUMN ULTRA triple quadrupole mass spectrometer).
2、实验方法2. Experimental method
称取化合物溶于DMSO-PEG-400-生理盐水(5：60：35，v/v/v)体系中，大鼠/猴静脉或灌胃给药后，于5min、15min、30min、1h、2h、4h、6h、8h、24h采集静脉血200μL于肝素化EP管中，12000rpm离心2min，取血浆-80℃冻存待测。 精密称取一定量供试品用DMSO溶解至1mg/mL，作为储备液。准确吸取适量的化合物储备液，加入乙腈稀释制成标准系列溶液。准确吸取上述标准系列溶液各20μL，加入空白血浆180μL，涡旋混匀，配制成相当于血浆浓度为1、3、10、30、100、300、1000、3000和5000ng/mL的血浆样品，每一浓度进行双样本分析，建立标准曲线。取20μL血浆，加入内标普萘洛尔(5ng/mL)的乙腈溶液200μL，涡旋混匀后4000rpm离心5min，取上清LC-MS分析。LC-MS检测条件如下：Weigh the compound and dissolve it in a DMSO-PEG-400-physiological saline (5:60:35, v/v/v) system. After intravenous or intragastric administration in rats/monkeys, at 5min, 15min, 30min, 1h, 200μL of venous blood was collected at 2h, 4h, 6h, 8h, 24h in a heparinized EP tube, centrifuged at 12000rpm for 2min, and the plasma was frozen at -80°C for testing. Accurately weigh a certain amount of the test substance and dissolve it to 1 mg/mL in DMSO as a stock solution. Accurately draw an appropriate amount of compound stock solution, add acetonitrile and dilute to prepare a standard series solution. Accurately draw 20μL of each of the above-mentioned standard series solutions, add 180μL of blank plasma, vortex to mix, and prepare plasma samples equivalent to plasma concentrations of 1, 3, 10, 30, 100, 300, 1000, 3000, and 5000 ng/mL. Perform two-sample analysis for one concentration and establish a standard curve. Take 20μL of plasma, add 200μL of internal standard propranolol (5ng/mL) acetonitrile solution, vortex to mix well, centrifuge at 4000rpm for 5min, take the supernatant for LC-MS analysis. The LC-MS detection conditions are as follows:
色谱柱：赛默飞HYPERSIL GOLD C-18 UPLC柱，100*2.1mm，1.9μm。Chromatographic column: Thermo Fisher HYPERSIL GOLD C-18 UPLC column, 100*2.1mm, 1.9μm.
流动相：水(0.1％甲酸)-乙腈按下表进行梯度洗脱Mobile phase: water (0.1% formic acid)-acetonitrile for gradient elution as shown in the table below
时间(min)Time (min) 水(含0.1％甲酸)Water (containing 0.1% formic acid) 乙腈Acetonitrile
00 90％90% 10％10%
0.60.6 90％90% 10％10%
11 10％10% 90％90%
2.62.6 10％10% 90％90%
2.612.61 90％90% 10％10%
44 90％90% 10％10%
3、数据处理3. Data processing
LC-MS检测血药浓度后，采用WinNonlin 6.1软件，非房室模型法计算药动学参数。结果见表三、四。After LC-MS detects the blood drug concentration, WinNonlin 6.1 software is used, and the non-compartmental model method is used to calculate the pharmacokinetic parameters. The results are shown in Tables 3 and 4.
表三本发明化合物的大鼠药代动力学参数(iv给药/0.5mg/kg)Table 3 Rat pharmacokinetic parameters of the compound of the present invention (iv administration/0.5mg/kg)
实施例Example Tmax(h)Tmax(h) AUC 0-∞(μg/L*h) AUC 0-∞ (μg/L*h) T1/2(h)T1/2(h) CL(L/h/kg)CL(L/h/kg) Vd(L/kg)Vd(L/kg) Cmax(μg/L)Cmax(μg/L)
88 0.0830.083 277.10277.10 0.680.68 1.801.80 1.781.78 525.07525.07
1313 0.0830.083 1093.061093.06 0.720.72 0.460.46 0.480.48 3123.263123.26
1414 0.0830.083 1444.081444.08 1.081.08 0.350.35 0.540.54 2703.732,073.73
1515 0.0830.083 674.62674.62 1.451.45 0.770.77 1.741.74 1340.941,340.94
1919 0.0830.083 1334.721334.72 0.520.52 0.380.38 0.290.29 2521.952521.95
21twenty one 0.0830.083 156.40156.40 0.230.23 3.313.31 1.081.08 445.06445.06
2525 0.0830.083 1260.731,260.73 0.270.27 0.400.40 0.160.16 3021.893021.89
2727 0.0830.083 863.50863.50 0.700.70 0.580.58 0.590.59 1041.011041.01
2828 0.0830.083 665.52665.52 0.800.80 0.750.75 0.880.88 1742.711,742.71
2929 0.0830.083 1327.671,327.67 0.730.73 0.380.38 0.390.39 3289.763,289.76
3131 0.0830.083 269.56269.56 0.460.46 2.002.00 1.381.38 527.25527.25
3232 0.0830.083 170.55170.55 0.170.17 2.982.98 0.760.76 474.46474.46
3535 0.0830.083 1110.291110.29 0.360.36 0.450.45 0.240.24 3089.623,089.62
4545 0.0830.083 120.90120.90 0.150.15 4.154.15 0.920.92 374.18374.18
4747 0.0830.083 126.79126.79 0.440.44 3.953.95 2.512.51 241.63241.63
5151 0.0830.083 3491.043,491.04 0.640.64 0.160.16 0.150.15 5949.975949.97
5353 0.0830.083 1682.591,682.59 0.720.72 0.300.30 0.310.31 2752.502,752.50
5555 0.0830.083 1358.781,358.78 0.420.42 0.370.37 0.220.22 2349.862,349.86
5656 0.0830.083 3925.343,925.34 1.101.10 0.130.13 0.210.21 4368.674,368.67
6161 0.0830.083 961.81961.81 0.890.89 0.520.52 0.670.67 850.92850.92
6666 0.0830.083 302.83302.83 0.340.34 1.651.65 0.820.82 1087.831087.83
表四本发明化合物的食蟹猴药代动力学参数(iv给药/1mg/kg)Table four cynomolgus monkey pharmacokinetic parameters of the compound of the present invention (iv administration/1mg/kg)
实施例Example Tmax(h)Tmax(h) T 1/2(h) T 1/2 (h) Cmax(μg/L)Cmax(μg/L) AUC 0-t(μg/L*h) AUC 0-t (μg/L*h) Vd(L/kg)Vd(L/kg) CL(L/h/kg)CL(L/h/kg)
2525 0.0830.083 1.541.54 5709.615,709.61 2057.962057.96 1.101.10 0.490.49
3131 0.0830.083 1.231.23 3619.643619.64 2128.152,128.15 0.830.83 0.470.47
4343 0.0830.083 2.502.50 2505.862505.86 1265.081265.08 2.722.72 0.790.79
5555 0.0830.083 1.081.08 43204320 14801480 0.320.32 0.700.70
3535 0.0830.083 0.710.71 40904090 13601360 0.340.34 0.740.74
21twenty one 0.0830.083 0.790.79 25502550 10301030 0.570.57 0.980.98
4747 0.0830.083 1.381.38 16001600 927927 1.11.1 1.071.07
结论：本发明化合物在大鼠和猴静注给药后半衰期较短，表观分布容积较低，清除率偏快。静滴给药后能迅速达稳，药物主要分布在血液中，靶向性较好，且停药后药物能迅速从体内清除，适合开发成临床静滴给药且主要靶点在血液中的药物。Conclusion: The compound of the present invention has a shorter half-life after intravenous administration in rats and monkeys, a lower apparent volume of distribution, and a faster clearance rate. After intravenous infusion, the drug can be quickly stabilized. The drug is mainly distributed in the blood, with good targeting, and the drug can be quickly removed from the body after the drug is stopped. It is suitable for development into clinical intravenous drug delivery and the main target is in the blood. drug.
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。The above-mentioned embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above-mentioned embodiments, and any other changes, modifications, substitutions, combinations, etc. made without departing from the spirit and principle of the present invention Simplified, all should be equivalent replacement methods, and they are all included in the protection scope of the present invention.

Claims (14)

  1. 式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐：The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts:
    Figure PCTCN2020130361-appb-100001
    Figure PCTCN2020130361-appb-100001
    R 1选自R 3取代或者未取代的四氮唑、R 3取代或者未取代的三氮唑； R 1 is selected from R 3 substituted or unsubstituted tetrazole, R 3 substituted or unsubstituted triazole;
    R 2选自R 4取代或者未取代的苯环，其中R 4选自-NR 5-(CH 2)n-CO-(CH 2)n-NR 6R 7、-NR 5-SO 2-NR 6R 7； R 2 is selected from R 4 substituted or unsubstituted benzene ring, wherein R 4 is selected from -NR 5 -(CH 2 )n-CO-(CH 2 )n-NR 6 R 7 , -NR 5 -SO 2 -NR 6 R 7 ;
    Ar选自至少一个R 8取代或者未取代的以下基团：
    Figure PCTCN2020130361-appb-100002
    Figure PCTCN2020130361-appb-100003
    Ar is selected from at least one of the following groups substituted or unsubstituted by R 8:
    Figure PCTCN2020130361-appb-100002
    Figure PCTCN2020130361-appb-100003
    其中，R 3选自氢、卤素、C 1-4的烷基、卤素取代的C 1-4的烷基； Wherein, R 3 is selected from hydrogen, halogen, C 1-4 alkyl, halogen substituted C 1-4 alkyl;
    R 5、R 6、R 7独立的选自氢、C 1-4的烷基、C 1-4的烷氧基-C 1-4烷基、-SO 2-C 1-4烷、-SO 2-苯、-C 1-6的一元或者二元醇、
    Figure PCTCN2020130361-appb-100004
    -(CH 2)n-C 3-12的脂肪环，或者其中NR 5与NR 6R 7之中的任意一个以上通过-(CH 2)n-成环；或者NR 6R 7一起构成C 3-12的脂肪环；前述C 3-12的脂肪环环上的一个以上碳原子被0-2个N、O、S原子所替代，所述脂肪环进一步被一个以上的R 9所取代；     R 5 , R 6 , and R 7 are independently selected from hydrogen, C 1-4 alkyl, C 1-4 alkoxy -C 1-4 alkyl, -SO 2 -C 1-4 alkane, -SO 2 -Benzene, -C 1-6 mono or dihydric alcohol,
    Figure PCTCN2020130361-appb-100004
    -(CH 2 )nC 3-12 aliphatic ring, or wherein any one or more of NR 5 and NR 6 R 7 form a ring through -(CH 2 )n-; or NR 6 R 7 together form C 3-12 The aliphatic ring; one or more carbon atoms on the aforementioned C 3-12 alicyclic ring are replaced by 0-2 N, O, S atoms, and the aliphatic ring is further replaced by more than one R 9 ;
    R 8选自氢、卤素、C 1-4的烷基、羟基、-C 1-4的羧酸、-C 1-4的羧酸-C 1-4醇酯； R 8 is selected from hydrogen, halogen, C 1-4 alkyl, hydroxyl, -C 1-4 carboxylic acid, -C 1-4 carboxylic acid-C 1-4 alcohol ester;
    R 9选自氢、-(CH 2)n-OH、-SO 2-C 1-4烷、-(CH 2)n-COOH、-酰胺、氰基、NR 10R 11-C 1-4的烷氧基、C 1-4的烷基、C 1-4的烷氧基、C 1-4的烷氧-C 1-4烷基、-CO-吗啉、-CO-NR 12-(CH 2)n-OH、HOOC-C 1-4烷氧基、
    Figure PCTCN2020130361-appb-100005
    R 9 is selected from hydrogen, -(CH 2 )n-OH, -SO 2 -C 1-4 alkane, -(CH 2 )n-COOH, -amide, cyano, NR 10 R 11 -C 1-4 Alkoxy, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 alkoxy-C 1-4 alkyl, -CO-morpholine, -CO-NR 12 -(CH 2 ) n-OH, HOOC-C 1-4 alkoxy,
    Figure PCTCN2020130361-appb-100005
    R 10、R 11、R 12独立选自氢或C 1-4烷基； R 10 , R 11 , and R 12 are independently selected from hydrogen or C 1-4 alkyl;
    前述n＝0-6的自然数。The aforementioned natural number of n=0-6.
  2. 根据权利要求1所述的式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其特征在于，-NR 5-(CH 2)n-CO-(CH 2)n-NR 6R 7包括
    Figure PCTCN2020130361-appb-100006
    Figure PCTCN2020130361-appb-100007
    其中，R 7和R 9如上定义。     The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts according to claim 1, wherein -NR 5 -(CH 2 )n-CO-( CH 2 )n-NR 6 R 7 includes
    Figure PCTCN2020130361-appb-100006
    Figure PCTCN2020130361-appb-100007
    Wherein, R 7 and R 9 are as defined above.
  3. 根据权利要求1所述的式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其特征在于，The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts according to claim 1, characterized in that:
    所述C 1-4的烷基选自甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基； The C 1-4 alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl;
    -C 1-4的羧酸选自甲酸、乙酸、丙酸、正丁酸、异丁酸、叔丁酸； -C 1-4 carboxylic acid is selected from formic acid, acetic acid, propionic acid, n-butyric acid, isobutyric acid, tert-butyric acid;
    -C 1-4的羧酸-C 1-4醇酯选自甲酸甲酯、甲酸乙酯、甲酸丙酯、甲酸丁酯、乙酸甲酯、乙酸乙酯、乙酸丙酯、乙酸丁酯、丙酸甲酯、丙酸乙酯、丙酸丙酯、丙酸丁酯、丁酸甲酯、丁酸乙酯、丁酸丙酯、丁酸丁酯。 -C 1-4 carboxylic acid-C 1-4 alcohol ester is selected from methyl formate, ethyl formate, propyl formate, butyl formate, methyl acetate, ethyl acetate, propyl acetate, butyl acetate, propyl Methyl acid, ethyl propionate, propyl propionate, butyl propionate, methyl butyrate, ethyl butyrate, propyl butyrate, butyl butyrate.
  4. 根据权利要求1所述的式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其特征在于，所述卤素选自氟、氯、溴、碘。The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts according to claim 1, wherein the halogen is selected from fluorine, chlorine, bromine, and iodine.
  5. 根据权利要求1所述的式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其特征在于，The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts according to claim 1, characterized in that:
    所述C 1-4的烷选自甲烷、乙烷、丙烷、异丙烷、正丁烷、异丁烷、仲丁烷、叔丁烷； The C 1-4 alkane is selected from methane, ethane, propane, isopropane, n-butane, isobutane, sec-butane, and tert-butane;
    所述C 1-4的烷氧基选自甲氧基、乙氧基、丙氧基、异丙氧基、正丁氧基、异丁氧基、仲丁氧基、叔丁氧基； The C 1-4 alkoxy group is selected from methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy;
    所述-(CH 2)n-OH选自羟基、甲醇、乙醇、正丙醇、正丁醇； The -(CH 2 )n-OH is selected from hydroxyl, methanol, ethanol, n-propanol, and n-butanol;
    所述-C 1-6的一元或者二元醇选自甲醇、乙醇、正丙醇、正丁醇、叔丁醇、1,3-丁二醇、3-甲基丁-1-醇、3-甲基戊-1-醇、4-甲基戊-1醇、3-甲基己-1醇、4-甲基己-1醇； The -C 1-6 monohydric or dihydric alcohol is selected from methanol, ethanol, n-propanol, n-butanol, tert-butanol, 1,3-butanediol, 3-methylbutan-1-ol, 3 -Methylpentan-1-ol, 4-methylpentan-1ol, 3-methylhexan-1ol, 4-methylhexan-1ol;
    所述NR 10R 11-C 1-4的烷氧基选自甲胺甲氧基、乙胺甲氧基、丙胺甲氧基、丁胺甲氧基、甲胺乙氧基、乙胺乙氧基、丙胺乙氧基、丁胺乙氧基、甲胺丙氧基、乙胺丙氧基、丙胺丙氧基、丁胺丙氧基、甲胺丁氧基、乙胺丁氧基、丙胺丁氧基、丁胺丁氧基、二甲胺基甲氧基、二甲胺基乙氧基、二甲胺基丙氧基、二甲胺基丁氧基； The alkoxy group of the NR 10 R 11 -C 1-4 is selected from the group consisting of methylamine methoxy, ethylamine methoxy, propylamine methoxy, butylamine methoxy, methylamine ethoxy, and ethylamine ethoxy. Propyl, propylamine ethoxy, butylamine ethoxy, methylamine propoxy, ethylamine propoxy, propylamine propoxy, butylamine propoxy, methylamine butoxy, ethylamine butoxy, propylamine Oxy, butylaminobutoxy, dimethylaminomethoxy, dimethylaminoethoxy, dimethylaminopropoxy, dimethylaminobutoxy;
    所述C 1-4的烷氧-C 1-4烷基选自甲氧甲基、乙氧甲基、丙氧甲基、丁氧甲基、甲氧乙基、乙氧乙基、丙氧乙基、丁氧乙基、甲氧丙基、乙氧丙基、丙氧丙基、丁氧丙基、甲氧丁基、乙氧丁基、丙氧丁基、丁氧丁基； The C 1-4 alkoxy-C 1-4 alkyl group is selected from methoxymethyl, ethoxymethyl, propoxymethyl, butoxymethyl, methoxyethyl, ethoxyethyl, propoxy Ethyl, butoxyethyl, methoxypropyl, ethoxypropyl, propoxypropyl, butoxypropyl, methoxybutyl, ethoxybutyl, propoxybutyl, butoxybutyl;
    所述-CO-NR 12-(CH 2)n-OH选自-CO-NH-CH 2OH、-CO-N(CH 3)-CH 2OH、-CO-NH-CH 2CH 2OH、-CO-N(CH 3)-CH 2CH 2OH； The -CO-NR 12 -(CH 2 )n-OH is selected from -CO-NH-CH 2 OH, -CO-N(CH 3 )-CH 2 OH, -CO-NH-CH 2 CH 2 OH, -CO-N(CH 3 )-CH 2 CH 2 OH;
    所述HOOC-C 1-4烷氧基选自HOOC-甲氧基、HOOC-乙氧基、HOOC-丙氧基、HOOC-异丙氧基、HOOC-正丁氧基、HOOC-异丁氧基、HOOC-仲丁氧基、HOOC-叔丁氧基。 The HOOC-C 1-4 alkoxy group is selected from HOOC-methoxy, HOOC-ethoxy, HOOC-propoxy, HOOC-isopropoxy, HOOC-n-butoxy, HOOC-isobutoxy Group, HOOC-sec-butoxy, HOOC-tert-butoxy.
  6. 根据权利要求1所述的式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其特征在于，所述C 3-12的脂肪环选自环丙烷、环丁烷、环戊烷、环己烷、环庚烷、环辛烷、环壬烷、环癸烷、螺[3,3]庚环、螺[3,5]壬环、螺[4,5]癸环； The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts according to claim 1, wherein the C 3-12 aliphatic ring is selected from cyclopropane , Cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, cyclodecane, spiro[3,3]heptane, spiro[3,5]nonane, spiro[4 ,5] decane ring;
    所述C 3-12的脂肪的一个以上碳原子被0-2个N、O、S原子所替代，选自：
    Figure PCTCN2020130361-appb-100008
    Figure PCTCN2020130361-appb-100009
    More than one carbon atom of the C 3-12 fat is replaced by 0-2 N, O, S atoms, selected from:
    Figure PCTCN2020130361-appb-100008
    Figure PCTCN2020130361-appb-100009
  7. 根据权利要求1所述的式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其特征在于，n＝0、1、2、3、4、5、6。The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts according to claim 1, wherein n=0,1,2,3,4,5 , 6.
  8. 根据权利要求1所述的式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其特征在于，The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts according to claim 1, characterized in that:
    R 1选自四氮唑、三氮唑； R 1 is selected from tetrazolium and triazole;
    R 2选自苯环、
    Figure PCTCN2020130361-appb-100010
    Figure PCTCN2020130361-appb-100011
    R 2 is selected from benzene ring,
    Figure PCTCN2020130361-appb-100010
    Figure PCTCN2020130361-appb-100011
    Figure PCTCN2020130361-appb-100012
    Figure PCTCN2020130361-appb-100012
    Ar选自以下基团：
    Figure PCTCN2020130361-appb-100013
    Figure PCTCN2020130361-appb-100014
    Ar is selected from the following groups:
    Figure PCTCN2020130361-appb-100013
    Figure PCTCN2020130361-appb-100014
  9. 根据权利要求1所述化合物，或其立体异构体、互变异构体、药学上可接受的盐，其特征在于，选自以下化合物：The compound according to claim 1, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, characterized in that it is selected from the following compounds:
    Figure PCTCN2020130361-appb-100015
    Figure PCTCN2020130361-appb-100015
    Figure PCTCN2020130361-appb-100016
    Figure PCTCN2020130361-appb-100016
    Figure PCTCN2020130361-appb-100017
    Figure PCTCN2020130361-appb-100017
    Figure PCTCN2020130361-appb-100018
    Figure PCTCN2020130361-appb-100018
    Figure PCTCN2020130361-appb-100019
    Figure PCTCN2020130361-appb-100019
    Figure PCTCN2020130361-appb-100020
    Figure PCTCN2020130361-appb-100020
    Figure PCTCN2020130361-appb-100021
    Figure PCTCN2020130361-appb-100021
    Figure PCTCN2020130361-appb-100022
    Figure PCTCN2020130361-appb-100022
    Figure PCTCN2020130361-appb-100023
    Figure PCTCN2020130361-appb-100023
  10. 根据权利要求1-9任一权利要求所述的式(I)的化合物或其立体异构体、互变异构体、药学上可接受的盐，其特征在于，所述药学上可接受的盐是指化合物与药学上可接受的酸或碱制备。The compound of formula (I) or its stereoisomers, tautomers, or pharmaceutically acceptable salts according to any one of claims 1-9, wherein the pharmaceutically acceptable Salt refers to a compound prepared with a pharmaceutically acceptable acid or base.
  11. 根据权利要求1-10任一项所述化合物，或其立体异构体、互变异构体、药学上可接受的盐，其特征在于：所述化合物的一个以上的氢原子上被同位素氘取代。The compound according to any one of claims 1-10, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, characterized in that: more than one hydrogen atom of the compound is deuterated by an isotope replace.
  12. 一种药物组合物，其特征在于，包括前述权利要求1-11任一项所述化合物，或其立体异构体、互变异构体、药学上可接受的盐和一种以上药学上可接受的载体，优选为口服或者注射给药的药物组合物。A pharmaceutical composition, characterized in that it comprises the compound of any one of the preceding claims 1-11, or its stereoisomers, tautomers, pharmaceutically acceptable salts and more than one pharmaceutically acceptable compound. The accepted carrier is preferably a pharmaceutical composition for oral or injection administration.
  13. 根据权利要求1-11任一项所述化合物，或其立体异构体、互变异构体、药学上可接受的盐在制备用于制备治疗FXIa相关疾病的药物用途，优选血栓相关疾病的药物用途。The use of the compound according to any one of claims 1-11, or its stereoisomers, tautomers, or pharmaceutically acceptable salts in the preparation of drugs for the treatment of FXIa-related diseases, preferably for thrombosis-related diseases Drug use.
  14. 根据权利要求12所述的药物组合物在制备用于制备治疗FXIa相关疾病的药物用途，优选血栓相关疾病的药物用途。The pharmaceutical composition according to claim 12 is used in the preparation of a medicine for the treatment of FXIa-related diseases, preferably a medicine for thrombosis-related diseases.
PCT/CN2020/130361 2019-11-21 2020-11-20 Dioxopiperazine derivative, preparation method therefore and pharmaceutical use thereof WO2021098817A1 (en)

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CN1101039A (en) * 1993-05-14 1995-04-05 第一制药株式会社 Piperazine derivatives
CN101218227A (en) * 2005-07-08 2008-07-09 阿斯利康(瑞典)有限公司 Heterocyclic sulfonamide derivatives as inhibitors of factor xa
CN102026996A (en) * 2008-03-13 2011-04-20 百时美施贵宝公司 Pyridazine derivatives as factor XIA inhibitors
WO2014009872A1 (en) * 2012-07-09 2014-01-16 Lupin Limited Tetrahydroquinazolinone derivatives as parp inhibitors

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