WO2021110076A1 - Oxamide derivatives, preparation method therefor and use thereof in medicine - Google Patents

Oxamide derivatives, preparation method therefor and use thereof in medicine Download PDF

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Publication number
WO2021110076A1
WO2021110076A1 PCT/CN2020/133493 CN2020133493W WO2021110076A1 WO 2021110076 A1 WO2021110076 A1 WO 2021110076A1 CN 2020133493 W CN2020133493 W CN 2020133493W WO 2021110076 A1 WO2021110076 A1 WO 2021110076A1
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Prior art keywords
phenyl
amino
mmol
tert
chloro
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PCT/CN2020/133493
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French (fr)
Chinese (zh)
Inventor
吴俊军
陆银锁
肖瑛
吕洋
周鹏
洪泽新
黄艺
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 CN202080079342.0A priority Critical patent/CN114728917B/en
Publication of WO2021110076A1 publication Critical patent/WO2021110076A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/56—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having carbon atoms of carboxamide groups bound to carbon atoms of carboxyl groups, e.g. oxamides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D257/00—Heterocyclic compounds containing rings having four nitrogen atoms as the only ring hetero atoms
    • C07D257/02—Heterocyclic compounds containing rings having four nitrogen atoms as the only ring hetero atoms not condensed with other rings
    • C07D257/04—Five-membered rings
    • C07D257/06—Five-membered rings with nitrogen atoms directly attached to the ring carbon atom

Definitions

  • the invention belongs to the technical field of chemical medicines, and relates to oxamide derivatives, their preparation methods and their applications 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 diseases 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 is continuously 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 thrombus.
  • Patent applications that disclose compounds with FXIa inhibitory activity include WO96/30396, WO99/41276, WO2013/093484, WO2004/002405, WO2013/056060, WO2017/005725, WO2017/023992, WO2018 /041122 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 oxamide derivatives, their preparation methods and their applications 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.
  • 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, -(CH 2 )n-CO-R 5 , 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 , -NR 5 -SO 2 -R 13 ;
  • 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, phenyl, C 1-4 alkyl, C 1-4 alkoxy-C 1-4 alkyl, -SO 2 -C 1-4 alkane , -SO 2 -benzene, -C 1-6 monohydric 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
  • 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 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, -CO-NR 10 , -CONR 10 R 11 , -CONR 10 -SO 2 -C 1-4 alkane, Cyano, -NR 10 R 11 , NR 10 R 11 -C 1-4 alkoxy, -C 1-4 carboxylic acid, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 alkoxy-C 1-4 alkoxy, -CO-morpholine, morpholine, -OC 1-6 mono or dihydric alcohol, HOOC-C 1-4 alkoxy,
  • R 10 , R 11 , and R 12 are independently selected from hydrogen, C 1-4 alkyl, -(CH 2 )n-OH;
  • R 13 is selected from C 1-4 alkyl substituted or unsubstituted phenyl
  • n 0-6.
  • -NR 5 -(CH 2 )n-CO-(CH 2 )n-NR 6 R 7 includes
  • R 7 and R 9 are as defined above.
  • the C 1-4 alkyl group is selected from methyl, ethyl, propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, Tert-butyl, cyclobutyl;
  • -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, cyclopropane, n-butane, isobutane, sec-butane, tert-butane, cyclopropane 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 group, methanol, ethanol, n-propanol, 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 NR 10 R 11 is selected from the group consisting of amino, methylamine, ethylamine, propylamine, butylamine, dimethylamine, methylethylamine, methylpropylamine, methylbutylamine, diethylamine, ethylenepropylamine, ethylbutylamine, dipropylamine, propylbutylamine Amine, dibutylamine.
  • 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 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 isotope deuterium.
  • Another object of the present invention provides 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 isotopic derivatization can generally prolong 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 examination 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 each 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 C Synthesis of methyl 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate
  • Step D Synthesis of 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid
  • Step F Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
  • Step G Synthesis of (S)-2-amino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step H Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
  • Step I Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl ) Propionic acid
  • Step J Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
  • Step K Synthesis of (S)-4-(2-amino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido) -1H-Indole-2-acid tert-butyl ester
  • Step L Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(3-Methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
  • Step M Synthesis of (S)-4-(2-(2-((((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
  • Step B Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
  • Step C Synthesis of (S)-2-amino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step D Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
  • Step E Synthesis of (S)-2-((Fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Propionic acid
  • Step F Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
  • Step G Synthesis of (S)-4-(2-amino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) -1H-Indole-2-acid tert-butyl ester
  • Step H Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
  • Step I Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
  • Step B Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine-1- (Yl)phenyl)tert-butyl propionate
  • Step C Synthesis of (S)-2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionic acid tert Butyl
  • Step D Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine-1- (Yl)phenyl)tert-butyl propionate
  • Step E Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine-1- (Yl)phenyl)propionic acid
  • Step F Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine -1-yl)phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
  • Step G Synthesis of (S)-4-(2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl) Propionamido)-1H-indole-2-acid tert-butyl ester
  • Step H Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
  • Step I Synthesis of (S)-4-(2-(2-((((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
  • Step B Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane Tert-butyl ester
  • Step C Synthesis of (S)-2-amino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step D Synthesis of (S)-2-((Fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane Tert-butyl ester
  • Step E Synthesis of (S)-2-((Fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane acid
  • Step F Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)benzene Yl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
  • Step G Synthesis of (S)-4-(2-amino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H -Indole-2-acid tert-butyl ester
  • Step H Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
  • Step I Synthesis of (S)-4-(2-(2-((((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
  • Step B Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
  • Step C Synthesis of (S)-2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
  • Step D Synthesis of (S)-2-((Fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
  • Step E Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Propionic acid
  • Step F Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
  • Step G Synthesis of (S)-4-(2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido) -1H-Indole-2-acid tert-butyl ester
  • Step H Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
  • Step I Synthesis of (S)-4-(2-(2-((((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
  • Step B Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(oxetan-3-yl)-2-oxopiperazine-1 -Yl)phenyl)propionamido)tert-butyl benzoate
  • Step C Synthesis of (S)-4-(2-amino-3-(4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamide Yl) tert-butyl benzoate
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
  • Step E Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
  • Step B Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine -1-yl)phenyl)propionamido)tert-butyl benzoate
  • Step C Synthesis of (S)-4-(2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl) (Propionamido) tert-butyl benzoate
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
  • Step E Synthesis of (S)-4-(2-(2-((((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
  • Step B Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido )Tert-Butyl Benzoate
  • Step C Synthesis of tert-butyl (S)-4-(2-amino-3-(4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)benzoate
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)tert-butyl benzoate
  • Step E Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)benzoic acid
  • Step A Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxo-8-oxa-2-azaspiro[4.5]decane- 2-yl)phenyl)propionamido)tert-butyl benzoate
  • Step B Synthesis of (S)-4-(2-amino-3-(4-(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propane Amido) tert-butyl benzoate
  • Step C Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4-(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)tert-butyl benzoate
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid
  • Step B Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazine- 1-yl)phenyl)propionamido)tert-butyl benzoate
  • Step C Synthesis of (S)-4-(2-amino-3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propane Amido) tert-butyl benzoate
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
  • Step E Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
  • Step A Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxo-2-azaspiro[4.5]decane-2-yl)benzene (Yl) propionamido) tert-butyl benzoate
  • Step B Synthesis of (S)-4-(2-amino-3-(4-(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid Tert-butyl ester
  • Step C Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4-(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)tert-butyl benzoate
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid
  • Step B Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(((tetrahydro-2H-pyran-4-yl)methyl)piperazine-1 -Yl)phenyl)propionamido)tert-butyl benzoate
  • Step C Synthesis of (S)-4-(2-amino-3-(4-(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamide Yl) tert-butyl benzoate
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamido) tert-butyl benzoate
  • Step E Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamido)benzoic acid
  • Step B Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)tert-butyl benzoate
  • Step C Synthesis of (S)-4-(2-amino-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) Tert-butyl benzoate
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
  • Step E Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
  • Step C Synthesis of methyl 2-((5,6-dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate
  • Step D Synthesis of 2-((5,6-Dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid
  • Step E Synthesis of (S)-4-(2-(2-(((5,6-Dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-phenylpropionamido) tert-butyl benzoate
  • Step F Synthesis of (S)-4-(2-(2-(((5,6-Dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-phenylpropionamido)benzoic acid
  • Step C Synthesis of methyl 2-((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetate
  • Step D Synthesis of 2-((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetic acid
  • Step E Synthesis of (S)-4-(2-(2-(((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) tert-butyl benzoate
  • Step F Synthesis of (S)-4-(2-(2-(((cyclopropylmethyl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid
  • Step A Synthesis of (S)-2-amino-N-(1-oxoisoindol-5-yl)-3-phenylpropionamide
  • Step B Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((3-oxoiso Indol-5-yl)amino)-3-phenylpropan-2-yl)oxalamide
  • Step A Synthesis of (S)-2-amino-N-(1-oxoisoindol-5-yl)-3-phenylpropionamide
  • Step B Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((1-oxoiso Indol-5-yl)amino)-3-phenylpropan-2-yl)oxalamide
  • Step A Synthesis of (S)-2-amino-N-(2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-3-phenylpropionamide
  • Step B Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((2-oxo- 2,3-Dihydro-1H-benzo[d]imidazol-5-yl)amino)-3-phenylprop-2-yl)oxamide
  • Step A Synthesis of methyl (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido)benzoate
  • Step B Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido ) Methyl benzoate
  • Step C Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)methyl benzoate
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
  • Step A Synthesis of tert-butyl (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(pyridin-3-yl)propionamido)benzoate
  • Step B Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(Pyridin-3-yl) propionamido) tert-butyl benzoate
  • Step C Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3- (Pyridin-3-yl)propionamido)benzoic acid
  • Step A Synthesis of (S)-5-(2-((tert-butoxycarbonyl)amino)-3-phenylpropionamido)-1H-indole-2-carboxylic acid ethyl ester
  • Step B Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid ethyl ester
  • Step C Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid
  • Step B Synthesis of tert-butyl 5-(4-nitrophenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylate
  • Step C Synthesis of tert-butyl 5-(4-aminophenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylate
  • Step D Synthesis of (S)-3-(4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide (Phenyl)-3-phenylpropionamido)phenyl)-5-oxo-2,5-dihydro-1H-pyrazole-1-carboxylic acid tert-butyl ester
  • Step E Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((4-(5- Oxo-2,5-dihydro-1H-pyrazol-3-yl)phenyl)amino)-3-phenylprop-2-yl)oxamide
  • Step A Synthesis of 2-((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetic acid
  • Step B Synthesis of (S)-4-(2-(2-(((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) tert-butyl benzoate
  • Step C Synthesis of (S)-4-(2-(2-((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetamido)-3-phenylpropane Amido) benzoic acid
  • Step C Synthesis of methyl 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetate
  • Step D Synthesis of 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetic acid
  • Step E Synthesis of (S)-4-(2-(2-((((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxy (Acetamido)-3-phenylpropionamido) tert-butyl benzoate
  • Step F Synthesis of (S)-4-(2-(2-((((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxy (Acetamido)-3-phenylpropionamido)benzoic acid
  • Step A Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(3-(4-(1H-imidazole-1-carboximido)phenyl)-2-(2-((5-chloro-2-(1H-tetra (Azol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Cyanopiperidine-1-carboximidamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step C Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino))-2-oxoacetamido) -3-(4-(4-Cyanopiperidine-1-carboximidylamino)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(((cyanoamino)(phenoxy)methyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step B Synthesis of (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(N',4-dicyanopiperidine-1-carboximidodiamino)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step C Synthesis of (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(N',4-dicyanopiperidine-1-carboximiddiamino)phenylpropionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of phenyl isocyanate
  • triphosgene (240.37 mg, 0.81 mmol) was added dropwise to ethyl acetate (5.0 ml) containing aniline (50.0 mg, 0.54 mmol), the dripping was completed, and the reaction was carried out at 80 degrees Celsius for 4 hours.
  • Step B Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-phenylureido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step C Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-phenylureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step C Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-oxopyrrolidin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step D Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-oxopyrrolidin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of 1-oxa-8-azaspiro[4.5]decane-3-ol trifluoroacetate
  • N-tert-butoxycarbonyl-1-oxa-8-azaspiro[4.5]decan-3-ol 100 mg, 0.39 mmol
  • dichloromethane 2.0 mL
  • Trifluoroacetic acid 1.0 mL
  • Step B Synthesis of 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step C Synthesis of 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate
  • tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate 700 mg, 2.3 mmol was added to dichloromethane (2.0 mL), and trifluoroacetic acid ( 1.0 ml), react at room temperature for 3 hours.
  • Step C Synthesis of (S)-5-(3-(4-(4-carbamoylpiperidine-1-carboxamido)phenyl)-2-(2-((5-chloro-2-(1H -Tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step D Synthesis of (S)-5-(3-(4-(4-carbamoylpiperidine-1-carboxamido)phenyl)-2-(2-((5-chloro-2-(1H -Tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate
  • tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate 1.5 g, 5.0 mmol was added to dichloromethane (2.0 mL), and trifluoroacetic acid ( 1.0 ml), react at room temperature for 3 hours.
  • Step C Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-((Methylsulfonyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step D Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-((methylsulfonyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of tert-butyl 4-(3-hydroxyazetidine-1-carbonyl)piperidine-1-carboxylate
  • Step B Synthesis of (3-hydroxyazetidin-1-yl)(piperidin-4-yl)methanone trifluoroacetate
  • tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate 1. g, 4.9 mmol
  • dichloromethane 2.0 mL
  • trifluoroacetic acid 1.0 ml
  • Step C Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(3-Hydroxyazetidine-1-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step D Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(3-Hydroxyazetidine-1-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid
  • reaction solution was quenched with 1M hydrochloric acid, adjusted to pH 3, diluted with ethyl acetate (200 ml), washed with water (30 ml ⁇ 2 times) and saturated brine (30 ml), and used for the organic phase After drying with anhydrous sodium sulfate, filtering and concentrating, the crude product (S)-3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid was directly used in the next reaction.
  • Step B Synthesis of tert-butyl (S)-4-(3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamidobenzoate
  • Step C Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamide Yl) tert-butyl benzoate
  • Step D Synthesis of tert-(S)-4-(2-amino-3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido tert-butyl)benzoic acid Butyl
  • Step E Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate
  • Step F Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(3-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
  • reaction solution was quenched with water (10 mL), the tetrahydrofuran was evaporated to dryness, the remaining aqueous solution was diluted with ethyl acetate (100 mL), and the mixture was washed with water (20 mL ⁇ 2 times) and saturated brine (20 mL). The phase was dried with anhydrous sodium sulfate, filtered and concentrated, and the obtained crude product was directly used in the next reaction.
  • Step B Synthesis of 4-chloro-2-amino-1-(2,2,2-trifluoroethoxy)benzene
  • reaction solution was 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, and the crude product obtained was used directly in the next step reaction.
  • Step C Synthesis of 2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetic acid
  • 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 the crude product and lithium hydroxide ( 1.0 g, 24.0 mmol) was dissolved in a mixed solution of tetrahydrofuran (30 mL)/water (10 mL), and reacted at room temperature for 5 hours.
  • Step D Synthesis of (S)-4-(2-(2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetamido )-3-Phenylpropionamido) tert-butyl benzoate
  • Step E Synthesis of (S)-4-(2-(2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetamido )-3-Phenylpropionamido)benzoic acid
  • Step A Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
  • Step A Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-morpholinylpiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
  • Step B Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-morpholinylpiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid

Abstract

The present invention belongs to the field of medicinal chemistry and relates to oxamide derivatives, a preparation method therefor and the use thereof in medicine. In particular, provided is a compound of formula (I) or a stereoisomer, a tautomer and a pharmaceutically acceptable salt thereof. These compounds are inhibitors of selective factor XIa (Factor XIa, called FXIa for short). The present invention also relates to a pharmaceutical composition comprising these compounds and the use of the compound in a drug for treating diseases such as thromboembolisms.

Description

草酰胺类衍生物、其制备方法及其在医药上的应用Oxamide derivatives, their preparation method and their application in medicine 技术领域Technical field
本发明属于化学药物技术领域，涉及草酰胺类衍生物、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 oxamide derivatives, their preparation methods and their applications 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 diseases 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 is continuously 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 the 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 Med 1991；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 Med 1991; 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WO96/30396、WO99/41276、WO2013/093484、WO2004/002405、WO2013/056060、WO2017/005725、WO2017/023992、WO2018/041122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 thrombus. Patent applications that disclose compounds with FXIa inhibitory activity include WO96/30396, WO99/41276, WO2013/093484, WO2004/002405, WO2013/056060, WO2017/005725, WO2017/023992, WO2018 /041122 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草酰胺类衍生物、其制备方法及其在医药上的应用。The invention provides a series of oxamide derivatives, their preparation methods and their applications in medicine.
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 PCTCN2020133493-appb-000001
Figure PCTCN2020133493-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 PCTCN2020133493-appb-000002
Figure PCTCN2020133493-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、-(CH 2)n-CO-R 5，其中R 4选自-NR 5-(CH 2)n-CO-(CH 2)n-NR 6R 7、
Figure PCTCN2020133493-appb-000003
-NR 5-SO 2-NR 6R 7、-NR 5-SO 2-R 13； R 2 is selected from R 4 substituted or unsubstituted benzene ring, -(CH 2 )n-CO-R 5 , wherein R 4 is selected from -NR 5 -(CH 2 )n-CO-(CH 2 )n-NR 6 R 7 ,
Figure PCTCN2020133493-appb-000003
-NR 5 -SO 2 -NR 6 R 7 , -NR 5 -SO 2 -R 13 ;
Ar选自至少一个R 8取代或者未取代的以下基团：
Figure PCTCN2020133493-appb-000004
Figure PCTCN2020133493-appb-000005
Ar is selected from at least one of the following groups substituted or unsubstituted by R 8:
Figure PCTCN2020133493-appb-000004
Figure PCTCN2020133493-appb-000005
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、苯基、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 PCTCN2020133493-appb-000006
-(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, phenyl, C 1-4 alkyl, C 1-4 alkoxy-C 1-4 alkyl, -SO 2 -C 1-4 alkane , -SO 2 -benzene, -C 1-6 monohydric or dihydric alcohol,
Figure PCTCN2020133493-appb-000006
-(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、-CO-NR 10、-CONR 10R 11、-CONR 10-SO 2-C 1-4烷、氰基、-NR 10R 11、NR 10R 11-C 1-4的烷氧基、-C 1-4的羧酸、C 1-4的烷基、C 1-4的烷氧基、C 1-4的烷氧-C 1-4烷氧基、-CO-吗啉、吗啉、-O-C 1-6的一元或者二元醇、HOOC-C 1-4烷氧基、
Figure PCTCN2020133493-appb-000007
R 9 is selected from hydrogen, -(CH 2 )n-OH, -SO 2 -C 1-4 alkane, -CO-NR 10 , -CONR 10 R 11 , -CONR 10 -SO 2 -C 1-4 alkane, Cyano, -NR 10 R 11 , NR 10 R 11 -C 1-4 alkoxy, -C 1-4 carboxylic acid, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 alkoxy-C 1-4 alkoxy, -CO-morpholine, morpholine, -OC 1-6 mono or dihydric alcohol, HOOC-C 1-4 alkoxy,
Figure PCTCN2020133493-appb-000007
R 10、R 11、R 12独立选自氢、C 1-4烷基、-(CH 2)n-OH； R 10 , R 11 , and R 12 are independently selected from hydrogen, C 1-4 alkyl, -(CH 2 )n-OH;
R 13选自C 1-4的烷基取代或者未取代的苯基； R 13 is selected from C 1-4 alkyl substituted or unsubstituted phenyl;
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，-NR 5-(CH 2)n-CO-(CH 2)n-NR 6R 7包括
Figure PCTCN2020133493-appb-000008
Figure PCTCN2020133493-appb-000009
As a preferred technical solution of the present invention, -NR 5 -(CH 2 )n-CO-(CH 2 )n-NR 6 R 7 includes
Figure PCTCN2020133493-appb-000008
Figure PCTCN2020133493-appb-000009
Figure PCTCN2020133493-appb-000010
包括
Figure PCTCN2020133493-appb-000011
Figure PCTCN2020133493-appb-000010
include
Figure PCTCN2020133493-appb-000011
其中，R 7和R 9如上定义。 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、环丙基、正丁基、异丁基、仲丁基、叔丁基、环丁基； As a preferred technical solution of the present invention, the C 1-4 alkyl group is selected from methyl, ethyl, propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, Tert-butyl, cyclobutyl;
-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、环丙烷、正丁烷、异丁烷、仲丁烷、叔丁烷、环丁烷； As a preferred technical solution of the present invention, the C 1-4 alkane is selected from methane, ethane, propane, isopropane, cyclopropane, n-butane, isobutane, sec-butane, tert-butane, cyclopropane 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 group, methanol, ethanol, n-propanol, 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选自氨基、甲胺、乙胺、丙胺、丁胺、二甲胺、甲乙胺、甲丙胺、甲丁胺、二乙胺、乙丙胺、乙丁胺、二丙胺、丙丁胺、二丁胺。 The NR 10 R 11 is selected from the group consisting of amino, methylamine, ethylamine, propylamine, butylamine, dimethylamine, methylethylamine, methylpropylamine, methylbutylamine, diethylamine, ethylenepropylamine, ethylbutylamine, dipropylamine, propylbutylamine Amine, dibutylamine.
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 PCTCN2020133493-appb-000012
Figure PCTCN2020133493-appb-000013
More than one carbon atom of the C 3-12 fat is replaced by 0-2 N, O, S atoms, selected from:
Figure PCTCN2020133493-appb-000012
Figure PCTCN2020133493-appb-000013
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，As a preferred technical solution of the present invention,
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 PCTCN2020133493-appb-000014
Figure PCTCN2020133493-appb-000015
Figure PCTCN2020133493-appb-000016
Figure PCTCN2020133493-appb-000017
R 2 is selected from benzene ring,
Figure PCTCN2020133493-appb-000014
Figure PCTCN2020133493-appb-000015
Figure PCTCN2020133493-appb-000016
Figure PCTCN2020133493-appb-000017
Ar选自以下基团：
Figure PCTCN2020133493-appb-000018
Figure PCTCN2020133493-appb-000019
Ar is selected from the following groups:
Figure PCTCN2020133493-appb-000018
Figure PCTCN2020133493-appb-000019
作为本发明的一种优选技术方案，选自以下化合物：As a preferred technical solution of the present invention, it is selected from the following compounds:
Figure PCTCN2020133493-appb-000020
Figure PCTCN2020133493-appb-000020
Figure PCTCN2020133493-appb-000021
Figure PCTCN2020133493-appb-000021
Figure PCTCN2020133493-appb-000022
Figure PCTCN2020133493-appb-000022
Figure PCTCN2020133493-appb-000023
Figure PCTCN2020133493-appb-000023
Figure PCTCN2020133493-appb-000024
Figure PCTCN2020133493-appb-000024
Figure PCTCN2020133493-appb-000025
Figure PCTCN2020133493-appb-000025
Figure PCTCN2020133493-appb-000026
Figure PCTCN2020133493-appb-000026
Figure PCTCN2020133493-appb-000027
Figure PCTCN2020133493-appb-000027
Figure PCTCN2020133493-appb-000028
Figure PCTCN2020133493-appb-000028
作为本发明的一种优选技术方案，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 technical solution 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 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 provides 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 PCTCN2020133493-appb-000029
为连接键。 Said
Figure PCTCN2020133493-appb-000029
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 isotopic derivatization can generally prolong 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 examination 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 each 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 the situation where the event or condition occurs and the situation where 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 U3000HPLC DAD高效液相色谱仪。High performance liquid chromatography (HPLC) analysis uses Thermo U3000HPLC 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)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(3-Methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000030
Figure PCTCN2020133493-appb-000030
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-氯-2-(1H-四唑-1-基)硝基苯Step A: Synthesis of 5-chloro-2-(1H-tetrazol-1-yl)nitrobenzene
Figure PCTCN2020133493-appb-000031
Figure PCTCN2020133493-appb-000031
室温下，将5-氯-2-氟硝基苯(5.0克，28.5毫摩尔)、四氮唑(2.4克，34.2毫摩尔)和碳酸钾(5.9克，42.7毫摩尔)加入N,N-二甲基甲酰胺(10.0毫升)中，N 2保护下，80摄氏度反应3.0小时。 At room temperature, 5-chloro-2-fluoronitrobenzene (5.0 g, 28.5 mmol), tetrazolium (2.4 g, 34.2 mmol) and potassium carbonate (5.9 g, 42.7 mmol) were added to N,N- In dimethylformamide (10.0 ml), under the protection of N 2 , the reaction was conducted at 80 degrees Celsius for 3.0 hours.
反应结束，加入乙酸乙酯30毫升，有机相混合物用饱和食盐水(20毫升×3次)洗涤，然后用无水硫酸钠干燥。有机相减压浓缩所得残余物直接进行下一步反应。After the reaction was completed, 30 mL of ethyl acetate was added, and the organic phase mixture was washed with saturated brine (20 mL×3 times), and then dried over anhydrous sodium sulfate. The residue obtained by concentrating the organic phase under reduced pressure was directly used for the next reaction.
步骤B：合成5-氯-2-(1H-四唑-1-基)苯胺Step B: Synthesis of 5-chloro-2-(1H-tetrazol-1-yl)aniline
Figure PCTCN2020133493-appb-000032
Figure PCTCN2020133493-appb-000032
室温下，将5-氯-2-(1H-四唑-1-基)硝基苯粗品加入乙酸乙酯(50毫升)中，冰浴下，分批加入氯化亚锡二水合物(67.0克，300毫摩尔)，N 2保护下，室温反应过夜。 At room temperature, the crude 5-chloro-2-(1H-tetrazol-1-yl)nitrobenzene was added to ethyl acetate (50 ml). Under an ice bath, stannous chloride dihydrate (67.0 g, 300 mmol), under N 2, 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50毫升乙酸乙酯洗涤，滤液用乙酸乙酯(50毫升×3次)萃取，合并有机相，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，得到副产物，乙酸乙酯/正己烷＝1/2，得到产物).得到3.0克黄色油状液体5-氯-2-(1H-四唑-1-基)苯胺(收率：54.1)。LCMS:RT＝3.59min,[M+H] +＝196.09。 After the reaction was completed, it was quenched with saturated sodium bicarbonate aqueous solution, and excess sodium bicarbonate was added to adjust the pH to weakly alkaline. The filter cake was filtered with Celite pad and the filter cake was washed with 50 ml of ethyl acetate. The filtrate was washed with ethyl acetate ( 50 ml×3 times) extract, combine the organic phases, then dry over anhydrous sodium sulfate, and finally concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/5 to obtain a by-product, ethyl acetate/n-hexane=1/2 to obtain a product). 3.0 g of yellow oily liquid was obtained 5-chloro-2-(1H-tetrazol-1-yl)aniline (yield: 54.1). LCMS: RT = 3.59 min, [M+H] + = 196.09.
步骤C：合成2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000033
Figure PCTCN2020133493-appb-000033
N 2保护、冰浴下，向含有5-氯-2-(1H-四唑-1-基)苯胺(3.0克，15.4毫摩尔)和吡啶(2.4克，30.8毫摩尔)的二氯甲烷(50毫升)中滴加草酰氯单甲酯(2.3克，18.5毫摩尔)，滴毕，室温反应1.0小时。 Under N 2 protection and ice bath, add 5-chloro-2-(1H-tetrazol-1-yl)aniline (3.0 g, 15.4 mmol) and pyridine (2.4 g, 30.8 mmol) to dichloromethane ( 50 mL) was added dropwise to oxalyl chloride monomethyl ester (2.3 g, 18.5 mmol), after the dropping, the reaction was carried out at room temperature for 1.0 hour.
反应结束，有机相先用饱和食盐水(40毫升×3次)洗涤，然后用无水硫酸钠干燥。有机相减压浓缩，得到白色块状固体2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸甲酯，直接用于下一步反应。After the reaction was over, the organic phase was washed with saturated brine (40 ml×3 times), and then dried with anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure to obtain methyl 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate as a white solid, which was used directly in the next step reaction.
步骤D：合成2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸Step D: Synthesis of 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000034
Figure PCTCN2020133493-appb-000034
冰浴下，向含有2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸甲酯粗品的四氢呋喃(40毫升)中滴加氢氧化锂(1.3克，30.8毫摩尔)水溶液(20毫升)，滴毕，室温反应30分钟。Under an ice bath, add hydrogen dropwise to tetrahydrofuran (40 ml) containing crude methyl 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate Aqueous solution (20 ml) of lithium oxide (1.3 g, 30.8 mmol) was dripped and reacted at room temperature for 30 minutes.
反应结束，用稀盐酸水溶液(1.0摩尔/升)调pH至3，用滤纸过滤混合物，滤饼用蒸馏水(30毫升)洗涤，干燥后得到3.2克白色絮状固体2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸。无需纯化，直接用于下一步反应。LCMS:RT＝1.52min,[M-H] -＝266.01。 After the reaction, the pH was adjusted to 3 with a dilute aqueous hydrochloric acid solution (1.0 mol/L), the mixture was filtered with filter paper, the filter cake was washed with distilled water (30 ml), and after drying, 3.2 g of white flocculent solid 2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid. No purification is required and it is directly used in the next reaction. LCMS: RT = 1.52 min, [MH] - = 266.01.
步骤E：合成4-(2-甲氧基丙基)哌嗪-2-酮Step E: Synthesis of 4-(2-methoxypropyl)piperazin-2-one
Figure PCTCN2020133493-appb-000035
Figure PCTCN2020133493-appb-000035
室温下，将2-氧代哌嗪(1.0毫克，10.0毫摩尔)、2-乙氧基乙基-1-溴乙烷(1.7克，11.0毫摩尔)和碳酸钾(3.5克，25.0毫摩尔)加入乙腈(20毫升)中，氮气保护下，冷凝回流，加热80摄氏度过夜。At room temperature, 2-oxopiperazine (1.0 mg, 10.0 mmol), 2-ethoxyethyl-1-bromoethane (1.7 g, 11.0 mmol) and potassium carbonate (3.5 g, 25.0 mmol) ) Was added to acetonitrile (20 ml), under nitrogen protection, condensed to reflux, and heated at 80 degrees Celsius overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用20毫升二氯甲烷洗涤，合并有机相，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602.0毫克白色固体4-(3-甲氧基丙基)哌嗪-2-酮(收率：35.0％)。At the end of the reaction, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 20 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtained 602.0 mg of white solid 4-(3-methoxypropyl)piperazin-2-one (yield: 35.0%).
步骤F：合成(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step F: Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
Figure PCTCN2020133493-appb-000036
Figure PCTCN2020133493-appb-000036
氮气保护下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-溴-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(700.0毫克，1.8毫摩尔)、4-(2-乙氧基乙基)哌嗪-2-酮(602.0毫克，3.5毫摩尔)、碳酸铯(1.4克，4.4毫摩尔)、碘化亚铜(332.0毫克，1.8毫摩尔)和N,N’-二甲基乙二胺(308.0毫克，3.5毫摩尔)加入甲苯(15毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, the (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-bromo-2-oxypiperazin-1-yl)phenyl)propionic acid tert-butyl ester ( 700.0 mg, 1.8 mmol), 4-(2-ethoxyethyl)piperazin-2-one (602.0 mg, 3.5 mmol), cesium carbonate (1.4 g, 4.4 mmol), cuprous iodide ( 332.0 mg, 1.8 mmol) and N,N'-dimethylethylenediamine (308.0 mg, 3.5 mmol) were added to toluene (15 mL), replaced with nitrogen three times, condensed and refluxed and heated at 110 degrees Celsius to react overnight.
反应结束，加入乙酸乙酯(30毫升)，混合物用饱和食盐水(20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/9)。得到610.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：70.6％)。LCMS:RT＝3.07min,[M+H] +＝492.27。 After the reaction was completed, ethyl acetate (30 mL) was added, the mixture was washed with saturated brine (20 mL × 3 times), then the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography ( Eluent: methanol/dichloromethane=1/9). Obtained 610.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazine-1- (Yl)phenyl)tert-butyl propionate (yield: 70.6%). LCMS: RT = 3.07 min, [M+H] + =492.27.
步骤G：合成(S)-2-氨基-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step G: Synthesis of (S)-2-amino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020133493-appb-000037
Figure PCTCN2020133493-appb-000037
室温下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(610.0毫克，1.2毫摩尔)加入乙酸乙酯(16.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，4.0毫升)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate (610.0 mg, 1.2 mmol) was added to ethyl acetate (16.0 ml), and the ethyl acetate solution of hydrogen chloride (2.0 mol/L, 4.0 ml) was added dropwise with stirring. After the addition, nitrogen React overnight at room temperature under protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到327.0毫克白色固体(S)-2-氨基-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝3.07min,[M+H] +＝392.26。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 327.0 mg of white solid ( S)-2-amino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate, the crude product is used directly in the next step reaction. LCMS: RT = 3.07 min, [M+H] + =392.26.
步骤H：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step H: Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
Figure PCTCN2020133493-appb-000038
Figure PCTCN2020133493-appb-000038
室温下，将(S)-2-氨基-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(327.0毫克，0.8毫摩尔和碳酸钠(355.0毫克，3.4毫摩尔)加入四氢呋喃(3.0毫升)和水(3.0毫升)的混合溶剂中，搅拌下加入芴甲氧羰酰氯(259.0毫克，1.0毫摩尔)，氮气保护下室温反应3.0小时。At room temperature, (S)-2-amino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (327.0 Mg, 0.8 mmol and sodium carbonate (355.0 mg, 3.4 mmol) were added to a mixed solvent of tetrahydrofuran (3.0 mL) and water (3.0 mL), and fluorene methoxycarbonyl chloride (259.0 mg, 1.0 mmol) was added with stirring, React at room temperature for 3.0 hours under nitrogen protection.
反应结束，加入饱和氯化钠溶液(20毫升)，混合物用二氯甲烷(20毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/20)。得到336.0毫克白色固体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：65.5％)。LCMS:RT＝3.44min,[M+H] +＝614.30。 After the reaction was completed, saturated sodium chloride solution (20 mL) was added, and the mixture was extracted with dichloromethane (20 mL×3 times). After the organic phases were combined, they were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/20). Obtained 336.0 mg of white solid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)benzene Methyl) tert-butyl propionate (yield: 65.5%). LCMS: RT = 3.44 min, [M+H] + =614.30.
步骤I：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸Step I: Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl ) Propionic acid
Figure PCTCN2020133493-appb-000039
Figure PCTCN2020133493-appb-000039
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(336.0毫克，0.6毫摩尔)加入二氯甲烷(4.0毫升)中，搅拌下滴加三氟乙酸(2.0毫升)，滴加完毕后，氮气保护下室温反应1.0小时。At room temperature, (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl ) Tert-Butyl propionate (336.0 mg, 0.6 mmol) was added to dichloromethane (4.0 mL), trifluoroacetic acid (2.0 mL) was added dropwise with stirring, and after the addition was completed, reacted at room temperature for 1.0 hour under nitrogen protection.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，得到黄色油状液体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸，粗品直接用于下一步反应。LCMS:RT＝3.01min,[M+H] +＝558.16。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump to obtain a yellow oily liquid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxy) Propyl)-2-oxopiperazin-1-yl)phenyl)propionic acid, the crude product was directly used in the next reaction. LCMS: RT = 3.01 min, [M+H] + =558.16.
步骤J：合成(S)-4-(2-((芴甲氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯Step J: Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000040
Figure PCTCN2020133493-appb-000040
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸粗品和4-氨基-1-叔丁氧羰基-吲哚-2-酸叔丁酯加入N,N-二甲基甲酰胺(2.0毫升)中，搅拌下滴加N,N-二异丙基乙胺，调节pH值至碱性。然后加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(312.0毫克，0.8毫摩尔)和N,N-二甲基甲酰胺(142.0毫克，1.1毫摩尔)，于氮气保护下室温反应过夜。At room temperature, (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl ) Crude propionic acid and 4-amino-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester were added to N,N-dimethylformamide (2.0 ml), and N,N-dimethicone was added dropwise with stirring. Isopropylethylamine, adjust the pH to alkaline. Then add 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (312.0 mg, 0.8 mmol) and N,N-dimethylformaldehyde Amide (142.0 mg, 1.1 mmol) was reacted overnight at room temperature under nitrogen protection.
反应结束，加入乙酸乙酯(15毫升)，混合液用饱和食盐水(10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：甲醇/二氯甲烷＝1/20)。得到436.4毫克白色固体(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯(收率：90.9％)。LCMS:RT＝4.03min,[M+H] +＝872.55。 After the reaction was completed, ethyl acetate (15 mL) was added, and the mixture was washed with 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 = 1/20). Obtained 436.4 mg of white solid (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl ) Propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester (yield: 90.9%). LCMS: RT = 4.03 min, [M+H] + =872.55.
步骤K：合成(S)-4-(2-氨基-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step K: Synthesis of (S)-4-(2-amino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido) -1H-Indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000041
Figure PCTCN2020133493-appb-000041
室温下，将(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯(436.4毫克，0.5毫摩尔)加入二氯甲烷(6.0毫升)中，搅拌下滴加4-甲基哌啶(1.5毫升)，于氮气保护下室温反应1.0小时。At room temperature, (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl) Propanamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester (436.4 mg, 0.5 mmol) was added to dichloromethane (6.0 mL), and 4-methylpiperidine ( 1.5 ml), and react at room temperature for 1.0 hour under the protection of nitrogen.
反应结束，直接减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/20)。得到186.2毫克白色固体(S)-4-(2-氨基-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯(收率：68.1％)。LCMS:RT＝2.89min,[M+H] +＝650.49。 After the reaction was over, it was directly concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/20). Obtain 186.2 mg of white solid (S)-4-(2-amino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido ) -1H-Indole-2-acid tert-butyl ester (yield: 68.1%). LCMS: RT = 2.89 min, [M+H] + =650.49.
步骤L：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step L: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(3-Methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000042
Figure PCTCN2020133493-appb-000042
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(220.0毫克，0.8毫摩尔)、(S)-4-(2-氨基-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯(186.2毫克，0.4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(390.0毫克，1.0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(175.0毫克，1.4毫摩尔)，滴毕，N 2保护下，室温反应1.0小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (220.0 mg, 0.8 mmol), (S)-4-( 2-amino-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl Ester (186.2 mg, 0.4 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (390.0 mg, 1.0 mmol) were added To N,N-dimethylformamide (4.0 mL), N,N-diisopropylethylamine (175.0 mg, 1.4 mmol) was added dropwise, after the dripping, the reaction was carried out at room temperature for 1.0 hour under the protection of N 2.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得粗品直接用于下一步反应。LCMS:RT＝3.26min,[M+H] +＝799.23。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained crude product was directly used in the next reaction. LCMS: RT = 3.26 min, [M+H] + =799.23.
步骤M：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Step M: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000043
Figure PCTCN2020133493-appb-000043
室温下，将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯粗品加入二氯甲烷(10.0毫升)中，滴加三氟乙酸(5.0毫升)，室温反应1.0小时。At room temperature, (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(3-Methoxypropyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl crude product was added to two Trifluoroacetic acid (5.0 mL) was added dropwise to methyl chloride (10.0 mL), and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到82.9毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸。LCMS:RT＝2.86min,[M+H] +＝743.29。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is separated by a preparative liquid chromatograph to obtain 82.9 mg of white solid (S)-4-(2-(2-(((5-chloro -2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-(3-methoxypropyl)-2-oxopiper (Azin-1-yl)phenyl)propionamido)-1H-indole-2-acid. LCMS: RT = 2.86 min, [M+H] + =743.29.
实施例2Example 2
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(2-Ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000044
Figure PCTCN2020133493-appb-000044
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(2-乙氧基乙基)哌嗪-2-酮Step A: Synthesis of 4-(2-ethoxyethyl)piperazin-2-one
Figure PCTCN2020133493-appb-000045
Figure PCTCN2020133493-appb-000045
室温下，将2-氧代哌嗪(1.0毫克，10.0毫摩尔)、3-甲氧基-1-溴乙烷(1.7克，11.0毫摩尔)和碳酸钾(3.5克，25.0毫摩尔)加入乙腈(20毫升)中，氮气保护下，冷凝回流，加热80摄氏度过夜。At room temperature, add 2-oxopiperazine (1.0 mg, 10.0 mmol), 3-methoxy-1-bromoethane (1.7 g, 11.0 mmol) and potassium carbonate (3.5 g, 25.0 mmol) Condensate to reflux in acetonitrile (20 ml) under nitrogen protection, and heat at 80 degrees Celsius overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用20毫升二氯甲烷洗涤，合并有机相，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668.0毫克白色固体4-(3-甲氧基丙基)哌嗪-2-酮(收率：38.9％)。At the end of the reaction, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 20 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). 668.0 mg of white solid 4-(3-methoxypropyl)piperazin-2-one was obtained (yield: 38.9%).
步骤B：合成(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step B: Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
Figure PCTCN2020133493-appb-000046
Figure PCTCN2020133493-appb-000046
氮气保护下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-溴-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(700.0毫克，1.8毫摩尔)、4-(2-乙氧基乙基)哌嗪-2-酮(668.0毫克，3.8毫摩尔)、碳酸铯(1.4克，4.4毫摩尔)、碘化亚铜(369.0毫克，1.8毫摩尔)和N,N’-二甲基乙二胺(343.0毫克，3.5毫摩尔)加入甲苯(15毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, the (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-bromo-2-oxypiperazin-1-yl)phenyl)propionic acid tert-butyl ester ( 700.0 mg, 1.8 mmol), 4-(2-ethoxyethyl)piperazin-2-one (668.0 mg, 3.8 mmol), cesium carbonate (1.4 g, 4.4 mmol), cuprous iodide ( 369.0 mg, 1.8 mmol) and N,N'-dimethylethylenediamine (343.0 mg, 3.5 mmol) were added to toluene (15 mL), replaced with nitrogen three times, and heated at 110 degrees Celsius under reflux for condensation and reacted overnight.
反应结束，加入乙酸乙酯(30毫升)，混合物用饱和食盐水(20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到663.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：77.6％)。LCMS:RT＝3.17min,[M+Na] +＝514.21。 After the reaction was completed, ethyl acetate (30 mL) was added, the mixture was washed with saturated brine (20 mL × 3 times), then the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography ( Eluent: methanol/dichloromethane=1/10). Obtain 663.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazine-1- (Yl)phenyl)tert-butyl propionate (yield: 77.6%). LCMS: RT = 3.17 min, [M+Na] + =514.21.
步骤C：合成(S)-2-氨基-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-amino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020133493-appb-000047
Figure PCTCN2020133493-appb-000047
室温下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(3-甲氧基丙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(663.0毫克，1.4毫摩尔)加入乙酸乙酯(16.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，4.0毫升)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(3-methoxypropyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate (663.0 mg, 1.4 mmol) was added to ethyl acetate (16.0 ml), and the ethyl acetate solution of hydrogen chloride (2.0 mol/L, 4.0 ml) was added dropwise with stirring. After the addition, nitrogen React overnight at room temperature under protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到217.0毫克白色固体(S)-2-氨基-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯，粗品直接用于下一步反应。After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 217.0 mg of white solid ( S)-2-amino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate, the crude product is used directly in the next step reaction.
步骤D：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
Figure PCTCN2020133493-appb-000048
Figure PCTCN2020133493-appb-000048
室温下，将(S)-2-氨基-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(217.0毫克，0.4毫摩尔)和碳酸钠(148.0毫克，1.4毫摩尔)加入四氢呋喃(3.0毫升)和水(3.0毫升)的混合溶剂中，搅拌下加入芴甲氧羰酰氯(122.0毫克，0.4毫摩尔)，氮气保护下室温反应3.0小时。At room temperature, (S)-2-amino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (217.0 Mg, 0.4 mmol) and sodium carbonate (148.0 mg, 1.4 mmol) were added to a mixed solvent of tetrahydrofuran (3.0 mL) and water (3.0 mL), and fluorene methoxycarbonyl chloride (122.0 mg, 0.4 mmol) was added with stirring , React at room temperature for 3.0 hours under nitrogen protection.
反应结束，加入饱和氯化钠溶液(20毫升)，混合物用二氯甲烷(20毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到343.0毫克白色固体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：100.0％)。LCMS:RT＝3.64min,[M+H] +＝614.30。 After the reaction was completed, saturated sodium chloride solution (20 mL) was added, and the mixture was extracted with dichloromethane (20 mL×3 times). After the organic phases were combined, they were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtain 343.0 mg of white solid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)benzene Yl) tert-butyl propionate (yield: 100.0%). LCMS: RT = 3.64 min, [M+H] + =614.30.
步骤E：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸Step E: Synthesis of (S)-2-((Fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Propionic acid
Figure PCTCN2020133493-appb-000049
Figure PCTCN2020133493-appb-000049
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(343.0毫克，0.4毫摩尔)加入二氯甲烷(4.0毫升)中，搅拌下滴加三氟乙酸(2.0毫升)，滴加完毕后，氮气保护下室温反应1.0小时。At room temperature, (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Tert-Butyl propionate (343.0 mg, 0.4 mmol) was added to dichloromethane (4.0 mL), trifluoroacetic acid (2.0 mL) was added dropwise under stirring, and after the addition was completed, reacted at room temperature for 1.0 hour under nitrogen protection.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，得到黄色油状液体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸，粗品直接用于下一步反应。LCMS:RT＝3.14min,[M+H] +＝558.24。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump to obtain a yellow oily liquid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxy) Ethyl)-2-oxopiperazin-1-yl)phenyl)propionic acid, the crude product was directly used in the next reaction. LCMS: RT = 3.14 min, [M+H] + =558.24.
步骤F：合成(S)-4-(2-((芴甲氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯Step F: Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000050
Figure PCTCN2020133493-appb-000050
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酸粗品和4-氨基-1-叔丁氧羰基-吲哚-2-酸叔丁酯加入N,N-二甲基甲酰胺(2.0毫升)中，搅拌下滴加N,N-二异丙基乙胺，调节pH值至碱性。然后加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(251.0毫克，0.7毫摩尔)和N,N-二甲基甲酰胺(170.0毫克，1.3毫摩尔)，于氮气保护下室温反应过夜。At room temperature, (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Crude propionic acid and 4-amino-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester were added to N,N-dimethylformamide (2.0 ml), and N,N-dimethicone was added dropwise with stirring. Isopropylethylamine, adjust the pH to alkaline. Then add 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (251.0 mg, 0.7 mmol) and N,N-dimethylformaldehyde Amide (170.0 mg, 1.3 mmol) was reacted at room temperature overnight under nitrogen protection.
反应结束，加入乙酸乙酯(15毫升)，混合液用饱和食盐水(10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)。得到221.0毫克白色固体(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯(收率：43.6％)。LCMS:RT＝4.17min,[M+H] +＝872.45。 After the reaction was completed, ethyl acetate (15 mL) was added, and the mixture was washed with 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). Obtain 221.0 mg of white solid (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl ) Propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester (yield: 43.6%). LCMS: RT = 4.17 min, [M+H] + =872.45.
步骤G：合成(S)-4-(2-氨基-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step G: Synthesis of (S)-4-(2-amino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) -1H-Indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000051
Figure PCTCN2020133493-appb-000051
室温下，将(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯(221.0毫克，0.2毫摩尔)加入二氯甲烷(6.0毫升)中，搅拌下滴加4-甲基哌啶(1.5毫升)，于氮气保护下室温反应1.0小时。At room temperature, (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl) Propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester (221.0 mg, 0.2 mmol) was added to dichloromethane (6.0 ml), and 4-methylpiperidine ( 1.5 ml), and react at room temperature for 1.0 hour under the protection of nitrogen.
反应结束，直接减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯)。得到147.0毫克白色固体(S)-4-(2-氨基-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯(收率：89.0％)。LCMS:RT＝2.94min,[M+H] +＝550.28。 After the reaction was over, it was directly concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate). Obtain 147.0 mg of white solid (S)-4-(2-amino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido ) -1H-Indole-2-acid tert-butyl ester (yield: 89.0%). LCMS: RT = 2.94 min, [M+H] + =550.28.
步骤H：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step H: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000052
Figure PCTCN2020133493-appb-000052
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(125.0毫克，0.5毫摩尔)、(S)-4-(2-氨基-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯(147.0毫克，0.2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(175.0毫克，0.5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(75.0毫克，0.6毫摩尔)，滴毕，N 2保护下，室温反应1.0小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (125.0 mg, 0.5 mmol), (S)-4-( 2-amino-3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl Ester (147.0 mg, 0.2 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (175.0 mg, 0.5 mmol) were added To N,N-dimethylformamide (4.0 mL), N,N-diisopropylethylamine (75.0 mg, 0.6 mmol) was added dropwise, after the dripping, the reaction was carried out at room temperature for 1.0 hour under the protection of N 2.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得粗品直接用于下一步反应。LCMS:RT＝3.32min,[M+H] +＝799.33。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained crude product was directly used in the next reaction. LCMS: RT = 3.32 min, [M+H] + =799.33.
步骤I：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Step I: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000053
Figure PCTCN2020133493-appb-000053
室温下，将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯粗品加入二氯甲烷(10.0毫升)中，滴加三氟乙酸(5.0毫升)，室温反应1.0小时。At room temperature, (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-Ethoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl crude product was added to two Trifluoroacetic acid (5.0 mL) was added dropwise to methyl chloride (10.0 mL), and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到43.0毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-乙氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸。LCMS:RT＝2.93min,[M+H] +＝743.10。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is separated by a preparative liquid chromatograph to obtain 43.0 mg of white solid (S)-4-(2-(2-(((5-chloro -2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-(2-ethoxyethyl)-2-oxopiper (Azin-1-yl)phenyl)propionamido)-1H-indole-2-acid. LCMS: RT = 2.93 min, [M+H] + =743.10.
实施例3Example 3
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(Tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000054
Figure PCTCN2020133493-appb-000054
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)-2-氧代哌嗪Step A: Synthesis of 4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine
Figure PCTCN2020133493-appb-000055
Figure PCTCN2020133493-appb-000055
室温下，将2-氧代哌嗪(1.0克，10.0毫摩尔)和四氢-2H-吡喃-4-酮(1.6克，16.0毫摩尔)加入甲醇(20毫升)和乙酸(0.4毫升)的混合溶剂中，氮气保护下，室温搅拌1.0小时。然后在冰浴下，缓慢加入氰基硼氢化钠(800.0毫克，13.0毫摩尔)，升至室温后，反应过夜。At room temperature, add 2-oxopiperazine (1.0 g, 10.0 mmol) and tetrahydro-2H-pyran-4-one (1.6 g, 16.0 mmol) to methanol (20 mL) and acetic acid (0.4 mL) In the mixed solvent, under the protection of nitrogen, stir at room temperature for 1.0 hour. Then in an ice bath, sodium cyanoborohydride (800.0 mg, 13.0 mmol) was slowly added, and after the temperature was raised to room temperature, the reaction was carried out overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用40毫升二氯甲烷洗涤，合并有机相，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1.5克白色固体4-(四氢-2H-吡喃-4-基)-2-氧代哌嗪(收率：89.6％)。After the reaction was completed, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 40 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 1.5 g of white solid 4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine was obtained (yield: 89.6%).
步骤B：合成(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step B: Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine-1- (Yl)phenyl)tert-butyl propionate
Figure PCTCN2020133493-appb-000056
Figure PCTCN2020133493-appb-000056
氮气保护下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-溴-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(500.0毫克，1.3毫摩尔)、4-(四氢-2H-吡喃-4-基)哌嗪-2-酮(460.0毫克，2.5毫摩尔)、碳酸铯(1.1克，3.1毫摩尔)、碘化亚铜(238.0毫克，1.3毫摩尔)和N,N’-二甲基乙二胺(220.0毫克，2.5毫摩尔)加入甲苯(15毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, the (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-bromo-2-oxypiperazin-1-yl)phenyl)propionic acid tert-butyl ester ( 500.0 mg, 1.3 mmol), 4-(tetrahydro-2H-pyran-4-yl)piperazin-2-one (460.0 mg, 2.5 mmol), cesium carbonate (1.1 g, 3.1 mmol), iodine Cuprous sulfide (238.0 mg, 1.3 mmol) and N,N'-dimethylethylenediamine (220.0 mg, 2.5 mmol) were added to toluene (15 mL), replaced with nitrogen three times, condensed and refluxed and heated at 110 degrees Celsius for reaction overnight.
反应结束，加入乙酸乙酯(30毫升)，混合物用饱和食盐水(20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/9)。得到371.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：58.9％)。LCMS:RT＝3.07min,[M+Na] +＝526.17。 After the reaction was completed, ethyl acetate (30 mL) was added, the mixture was washed with saturated brine (20 mL × 3 times), and then the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography ( Eluent: methanol/dichloromethane=1/9). Obtain 371.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxypiper (Azin-1-yl)phenyl) tert-butyl propionate (yield: 58.9%). LCMS: RT = 3.07 min, [M+Na] + =526.17.
步骤C：合成(S)-2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionic acid tert Butyl
Figure PCTCN2020133493-appb-000057
Figure PCTCN2020133493-appb-000057
室温下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(371.0毫克，0.8毫摩尔)加入乙酸乙酯(16.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，4.0毫升)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxypiperazine-1- (Phenyl) phenyl) tert-butyl propionate (371.0 mg, 0.8 mmol) was added to ethyl acetate (16.0 mL), and ethyl acetate solution of hydrogen chloride (2.0 mol/L, 4.0 mL) was added dropwise with stirring. After completion, react overnight at room temperature under nitrogen protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到217.0毫克白色固体(S)-2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝0.56min,[M+H] +＝426.16。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 217.0 mg of white solid ( S)-2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate, crude product directly Used in the next reaction. LCMS: RT=0.56 min, [M+H] + =426.16.
步骤D：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine-1- (Yl)phenyl)tert-butyl propionate
Figure PCTCN2020133493-appb-000058
Figure PCTCN2020133493-appb-000058
室温下，将(S)-2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(197.0毫克，0.5毫摩尔)和碳酸钠(212.0毫克，2.0毫摩尔)加入四氢呋喃(3.0毫升)和水(3.0毫升)的混合溶剂中，搅拌下加入芴甲氧羰酰氯(155.0毫克，0.6毫摩尔)，氮气保护下室温反应3.0小时。At room temperature, the (S)-2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionic acid tert Butyl ester (197.0 mg, 0.5 mmol) and sodium carbonate (212.0 mg, 2.0 mmol) were added to a mixed solvent of tetrahydrofuran (3.0 mL) and water (3.0 mL), and fluorenyl methoxycarbonyl chloride (155.0 mg, 0.6 mmol), and react at room temperature for 3.0 hours under the protection of nitrogen.
反应结束，加入饱和氯化钠溶液(20毫升)，混合物用二氯甲烷(20毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/9)。得到224.0毫克白色固体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：73.2％)。LCMS:RT＝3.60min,[M+H] +＝626.28。 After the reaction was completed, saturated sodium chloride solution (20 mL) was added, and the mixture was extracted with dichloromethane (20 mL×3 times). After the organic phases were combined, they were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtain 224.0 mg of white solid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxypiperazine-1 -Yl)phenyl)tert-butyl propionate (yield: 73.2%). LCMS: RT = 3.60 min, [M+H] + =626.28.
步骤E：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸Step E: Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine-1- (Yl)phenyl)propionic acid
Figure PCTCN2020133493-appb-000059
Figure PCTCN2020133493-appb-000059
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(224.0毫克，0.4毫摩尔)加入二氯甲烷(4.0毫升)中，搅拌下滴加三氟乙酸(2.0毫升)，滴加完毕后，氮气保护下室温反应1.0小时。At room temperature, the (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine-1- Tert-butyl phenyl)propionate (224.0 mg, 0.4 mmol) was added to dichloromethane (4.0 mL), trifluoroacetic acid (2.0 mL) was added dropwise with stirring, after the addition, the reaction was carried out at room temperature under nitrogen protection 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，得到黄色油状液体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸，粗品直接用于下一步反应。LCMS:RT＝3.03min,[M+H] +＝570.23。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump to obtain a yellow oily liquid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-) Pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionic acid, the crude product was directly used in the next reaction. LCMS: RT = 3.03 min, [M+H] + =570.23.
步骤F：合成(S)-4-(2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯Step F: Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine -1-yl)phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000060
Figure PCTCN2020133493-appb-000060
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酸粗品和4-氨基-1-叔丁氧羰基-吲哚-2-酸叔丁酯加入N,N-二甲基甲酰胺(2.0毫升)中，搅拌下滴加N,N-二异丙基乙胺，调节pH值至碱性。然后加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(205.0毫克，0.5毫摩尔)和N,N-二甲基甲酰胺(96.0毫克，0.7毫摩尔)，于氮气保护下室温反应过夜。At room temperature, the (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine-1- (Yl)phenyl)propionic acid crude product and 4-amino-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester were added to N,N-dimethylformamide (2.0 ml), and N was added dropwise with stirring. , N-Diisopropylethylamine, adjust the pH to alkaline. Then add 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (205.0 mg, 0.5 mmol) and N,N-dimethylformaldehyde Amide (96.0 mg, 0.7 mmol) was reacted at room temperature overnight under nitrogen protection.
反应结束，加入乙酸乙酯(15毫升)，混合液用饱和食盐水(10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：甲醇/二氯甲烷＝1/9)。得到95.0毫克白色固体(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯(收率：43.6％)。LCMS:RT＝4.17min,[M+H] +＝884.35。 After the reaction was completed, ethyl acetate (15 mL) was added, and the mixture was washed with 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=1/9). Obtain 95.0 mg of white solid (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxypiperazine-1- (Yl)phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester (yield: 43.6%). LCMS: RT = 4.17 min, [M+H] + =884.35.
步骤G：合成(S)-4-(2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step G: Synthesis of (S)-4-(2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl) Propionamido)-1H-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000061
Figure PCTCN2020133493-appb-000061
室温下，将(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯(95.0毫克，0.2毫摩尔)加入二氯甲烷(6.0毫升)中，搅拌下滴加4-甲基哌啶(1.5毫升)，于氮气保护下室温反应1.0小时。At room temperature, the (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl )Phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester (95.0 mg, 0.2 mmol) was added to dichloromethane (6.0 mL), and 4-methyl was added dropwise with stirring Piperidine (1.5 mL) was reacted at room temperature for 1.0 hour under the protection of nitrogen.
反应结束，直接减压浓缩。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130.0毫克白色固体(S)-4-(2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯。LCMS:RT＝1.42min,[M-H] -＝560.26。 After the reaction was over, it was directly concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtain 130.0 mg of white solid (S)-4-(2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl ) Propionamido) -1H-indole-2-acid tert-butyl ester. LCMS: RT=1.42 min, [MH] - =560.26.
步骤H：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step H: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000062
Figure PCTCN2020133493-appb-000062
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(75.0毫克，0.3毫摩尔)、(S)-4-(2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯(130.0毫克，0.2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(137.0毫克，0.4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(60.0毫克，0.5毫摩尔)，滴毕，N 2保护下，室温反应1.0小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (75.0 mg, 0.3 mmol), (S)-4-( 2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2 -Tert-butyl acid (130.0 mg, 0.2 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (137.0 mg, 0.4 Millimoles) was added to N,N-dimethylformamide (4.0 ml), and N,N-diisopropylethylamine (60.0 mg, 0.5 mmol) was added dropwise, after dripping, the reaction was carried out at room temperature under the protection of N 2 1.0 hour.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得粗品直接用于下一步反应。LCMS:RT＝3.26min,[M+H] +＝811.33。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained crude product was directly used in the next reaction. LCMS: RT = 3.26 min, [M+H] + =811.33.
步骤I：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Step I: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000063
Figure PCTCN2020133493-appb-000063
室温下，将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯粗品加入二氯甲烷(10.0毫升)中，滴加三氟乙酸(5.0毫升)，室温反应1.0小时。At room temperature, (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl The crude ester was added to dichloromethane (10.0 mL), trifluoroacetic acid (5.0 mL) was added dropwise, and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到43.0毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸。LCMS:RT＝2.86min,[M-H] -＝753.09。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is separated by a preparative liquid chromatograph to obtain 43.0 mg of white solid (S)-4-(2-(2-(((5-chloro -2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)- 2-Oxypiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid. LCMS: RT = 2.86 min, [MH] - =753.09.
实施例4Example 4
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(Tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000064
Figure PCTCN2020133493-appb-000064
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(四氢呋喃-3-基)-2-氧代哌嗪Step A: Synthesis of 4-(tetrahydrofuran-3-yl)-2-oxopiperazine
Figure PCTCN2020133493-appb-000065
Figure PCTCN2020133493-appb-000065
室温下，将2-氧代哌嗪(1.0克，10.0毫摩尔)和3-氧代四氢呋喃(1.3克，15.1毫摩尔)加入甲醇(20毫升)和乙酸(0.4毫升)的混合溶剂中，氮气保护下，室温搅拌1.0小时。然后在冰浴下，缓慢加入氰基硼氢化钠(800.0毫克，13.0毫摩尔)，升至室温后，反应过夜。At room temperature, 2-oxopiperazine (1.0 g, 10.0 mmol) and 3-oxotetrahydrofuran (1.3 g, 15.1 mmol) were added to a mixed solvent of methanol (20 mL) and acetic acid (0.4 mL). Nitrogen Under protection, stir at room temperature for 1.0 hour. Then in an ice bath, sodium cyanoborohydride (800.0 mg, 13.0 mmol) was slowly added, and after the temperature was raised to room temperature, the reaction was carried out overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用40毫升二氯甲烷洗涤，合并有机相，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819.0毫克白色固体4-(四氢呋喃-3-基)-2-氧代哌嗪(收率：51.2％)。After the reaction was completed, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 40 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 819.0 mg of white solid 4-(tetrahydrofuran-3-yl)-2-oxopiperazine was obtained (yield: 51.2%).
步骤B：合成(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step B: Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane Tert-butyl ester
Figure PCTCN2020133493-appb-000066
Figure PCTCN2020133493-appb-000066
氮气保护下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-溴-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(500.0毫克，1.3毫摩尔)、4-(四氢呋喃-3-基)哌嗪-2-酮(425.0毫克，2.5毫摩尔)、碳酸铯(1.1克，3.1毫摩尔)、碘化亚铜(238.0毫克，1.3毫摩尔)和N,N’-二甲基乙二胺(220.0毫克，2.5毫摩尔)加入甲苯(15毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, the (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-bromo-2-oxypiperazin-1-yl)phenyl)propionic acid tert-butyl ester ( 500.0 mg, 1.3 mmol), 4-(tetrahydrofuran-3-yl)piperazin-2-one (425.0 mg, 2.5 mmol), cesium carbonate (1.1 g, 3.1 mmol), cuprous iodide (238.0 mg , 1.3 mmol) and N,N'-dimethylethylenediamine (220.0 mg, 2.5 mmol) were added to toluene (15 mL), replaced with nitrogen three times, and heated at 110°C under reflux for condensation and reacted overnight.
反应结束，加入乙酸乙酯(30毫升)，混合物用饱和食盐水(20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/9)。得到538.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：86.0％)。LCMS:RT＝3.44min,[M+H] +＝512.15。 After the reaction was completed, ethyl acetate (30 mL) was added, the mixture was washed with saturated brine (20 mL × 3 times), and then the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography ( Eluent: methanol/dichloromethane=1/9). Obtained 538.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl) Phenyl) tert-butyl propionate (yield: 86.0%). LCMS: RT = 3.44 min, [M+H] + =512.15.
步骤C：合成(S)-2-氨基-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-amino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020133493-appb-000067
Figure PCTCN2020133493-appb-000067
室温下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(538.0毫克，0.8毫摩尔)加入乙酸乙酯(16.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，4.0毫升)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane Tert-Butyl acid (538.0 mg, 0.8 mmol) was added to ethyl acetate (16.0 ml), and ethyl acetate solution of hydrogen chloride (2.0 mol/L, 4.0 ml) was added dropwise with stirring. After the addition was completed, under nitrogen protection React overnight at room temperature.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到386.0毫克白色固体(S)-2-氨基-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝0.65min,[M+H] +＝412.73。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 386.0 mg of white solid ( S)-2-amino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate, the crude product was directly used in the next reaction. LCMS: RT=0.65 min, [M+H] + =412.73.
步骤D：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-((Fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane Tert-butyl ester
Figure PCTCN2020133493-appb-000068
Figure PCTCN2020133493-appb-000068
室温下，将(S)-2-氨基-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(386.0毫克，1.0毫摩尔)和碳酸钠(424.0毫克，4.0毫摩尔)加入四氢呋喃(4.0毫升)和水(4.0毫升)的混合溶剂中，搅拌下加入芴甲氧羰酰氯(310.0毫克，1.2毫摩尔)，氮气保护下室温反应3.0小时。At room temperature, (S)-2-amino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate (386.0 mg, 1.0 mmol) and sodium carbonate (424.0 mg, 4.0 mmol) were added to a mixed solvent of tetrahydrofuran (4.0 mL) and water (4.0 mL), fluorenyl methoxycarbonyl chloride (310.0 mg, 1.2 mmol) was added with stirring, and nitrogen was added. React at room temperature for 3.0 hours under protection.
反应结束，加入饱和氯化钠溶液(20毫升)，混合物用二氯甲烷(20毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/9)。得到391.0毫克白色固体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：64.3％)。LCMS:RT＝3.97min,[M+H] +＝634.29。 After the reaction was completed, saturated sodium chloride solution (20 mL) was added, and the mixture was extracted with dichloromethane (20 mL×3 times). After the organic phases were combined, they were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtain 391.0 mg of white solid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl) Tert-Butyl propionate (yield: 64.3%). LCMS: RT = 3.97 min, [M+H] + =634.29.
步骤E：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸Step E: Synthesis of (S)-2-((Fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane acid
Figure PCTCN2020133493-appb-000069
Figure PCTCN2020133493-appb-000069
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(391.0毫克，0.6毫摩尔)加入二氯甲烷(4.0毫升)中，搅拌下滴加三氟乙酸(2.0毫升)，滴加完毕后，氮气保护下室温反应1.0小时。At room temperature, (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane Tert-butyl ester (391.0 mg, 0.6 mmol) was added to dichloromethane (4.0 mL), trifluoroacetic acid (2.0 mL) was added dropwise with stirring, and after the addition was completed, the reaction was carried out at room temperature for 1.0 hour under nitrogen protection.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，得到黄色油状液体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸，粗品直接用于下一步反应。LCMS:RT＝3.35min,[M+H] +＝556.30。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump to obtain a yellow oily liquid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl) )-2-oxopiperazin-1-yl)phenyl)propionic acid, the crude product was directly used in the next reaction. LCMS: RT = 3.35 min, [M+H] + =556.30.
步骤F：合成(S)-4-(2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯Step F: Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)benzene Yl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000070
Figure PCTCN2020133493-appb-000070
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酸粗品和4-氨基-1-叔丁氧羰基-吲哚-2-酸叔丁酯加入N,N-二甲基甲酰胺(2.0毫升)中，搅拌下滴加N,N-二异丙基乙胺，调节pH值至碱性。然后加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(365.0毫克，1.0毫摩尔)和N,N-二甲基甲酰胺(165.0毫克，1.3毫摩尔)，于氮气保护下室温反应过夜。At room temperature, (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane The crude acid and 4-amino-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester were added to N,N-dimethylformamide (2.0 ml), and N,N-diisopropyl was added dropwise with stirring Ethylethylamine, adjust the pH to alkaline. Then add 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (365.0 mg, 1.0 mmol) and N,N-dimethylformaldehyde Amide (165.0 mg, 1.3 mmol) was reacted overnight at room temperature under nitrogen protection.
反应结束，加入乙酸乙酯(15毫升)，混合液用饱和食盐水(10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：甲醇/二氯甲烷＝1/9)。得到465.0毫克白色固体(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯(收率：94.5％)。LCMS:RT＝4.62min,[M+H] +＝871.25。 After the reaction was completed, ethyl acetate (15 mL) was added, and the mixture was washed with 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=1/9). Obtain 465.0 mg of white solid (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propane Amido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester (yield: 94.5%). LCMS: RT = 4.62 min, [M+H] + =871.25.
步骤G：合成(S)-4-(2-氨基-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step G: Synthesis of (S)-4-(2-amino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H -Indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000071
Figure PCTCN2020133493-appb-000071
室温下，将(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯(465.0毫克，0.8毫摩尔)加入二氯甲烷(6.0毫升)中，搅拌下滴加4-甲基哌啶(1.5毫升)，于氮气保护下室温反应1.0小时。At room temperature, (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamide Yl)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester (465.0 mg, 0.8 mmol) was added to dichloromethane (6.0 ml), and 4-methylpiperidine (1.5 ml ), react at room temperature for 1.0 hour under the protection of nitrogen.
反应结束，直接减压浓缩。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263.0毫克白色固体(S)-4-(2-氨基-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯。LCMS:RT＝2.65min,[M-H] -＝546.24。 After the reaction was over, it was directly concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtained 263.0 mg of white solid (S)-4-(2-amino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)- 1H-Indole-2-acid tert-butyl ester. LCMS: RT = 2.65 min, [MH] - =546.24.
步骤H：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step H: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000072
Figure PCTCN2020133493-appb-000072
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(155.0毫克，0.6毫摩尔)、(S)-4-(2-氨基-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯(263.0毫克，0.5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(274.0毫克，0.7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(124.0毫克，1.0毫摩尔)，滴毕，N 2保护下，室温反应1.0小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (155.0 mg, 0.6 mmol), (S)-4-( 2-amino-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester ( 263.0 mg, 0.5 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (274.0 mg, 0.7 mmol) add N, To N-dimethylformamide (4.0 mL), N,N-diisopropylethylamine (124.0 mg, 1.0 mmol) was added dropwise, and after the dripping was completed, the reaction was carried out at room temperature for 1.0 hour under the protection of N 2.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得粗品直接用于下一步反应。LCMS:RT＝3.62min,[M+H] +＝797.49。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained crude product was directly used in the next reaction. LCMS: RT=3.62min, [M+H] + =797.49.
步骤I：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Step I: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000073
Figure PCTCN2020133493-appb-000073
室温下，将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯粗品加入二氯甲烷(10.0毫升)中，滴加三氟乙酸(5.0毫升)，室温反应1.0小时。At room temperature, (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl crude product is added to dichloromethane (10.0 mL), trifluoroacetic acid (5.0 mL) was added dropwise, and reacted at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到288.0毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢呋喃-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸。LCMS:RT＝3.06min,[M-H] -＝739.12。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was separated by preparative liquid chromatography to obtain 288.0 mg of white solid (S)-4-(2-(2-(((5-chloro -2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-(tetrahydrofuran-3-yl)-2-oxopiperazine- 1-yl)phenyl)propionamido)-1H-indole-2-acid. LCMS: RT=3.06min, [MH] - =739.12.
实施例5Example 5
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000074
Figure PCTCN2020133493-appb-000074
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(4-甲氧基环己基)-2-氧代哌嗪Step A: Synthesis of 4-(4-methoxycyclohexyl)-2-oxopiperazine
Figure PCTCN2020133493-appb-000075
Figure PCTCN2020133493-appb-000075
室温下，将2-氧代哌嗪(900.0毫克，8.5毫摩尔)和4-甲氧基环己酮(2.2克，17.0毫摩尔)加入甲醇(40毫升)和乙酸(0.8毫升)的混合溶剂中，氮气保护下，室温搅拌1.0小时。然后在冰浴下，缓慢加入氰基硼氢化钠(1.2克，18.7毫摩尔)，升至室温后，反应过夜。At room temperature, add 2-oxopiperazine (900.0 mg, 8.5 mmol) and 4-methoxycyclohexanone (2.2 g, 17.0 mmol) to a mixed solvent of methanol (40 mL) and acetic acid (0.8 mL) Under the protection of nitrogen, stir at room temperature for 1.0 hour. Then in an ice bath, sodium cyanoborohydride (1.2 g, 18.7 mmol) was slowly added, and after the temperature was raised to room temperature, the reaction was carried out overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用40毫升二氯甲烷洗涤，合并有机相，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2.2克白色固体4-(4-甲氧基环己基)-2-氧代哌嗪。After the reaction was completed, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 40 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). 2.2 g of white solid 4-(4-methoxycyclohexyl)-2-oxopiperazine was obtained.
步骤B：合成(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step B: Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
Figure PCTCN2020133493-appb-000076
Figure PCTCN2020133493-appb-000076
氮气保护下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-溴-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(1.0克，2.5毫摩尔)、4-(4-甲氧基环己基)哌嗪-2-酮(1.0克，5.0毫摩尔)、碳酸铯(1.6克，5.0毫摩尔)、碘化亚铜(476.0毫克，5.0毫摩尔)和N,N’-二甲基乙二胺(440.0毫克，5.0毫摩尔)加入甲苯(15毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, the (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-bromo-2-oxypiperazin-1-yl)phenyl)propionic acid tert-butyl ester ( 1.0 g, 2.5 mmol), 4-(4-methoxycyclohexyl)piperazin-2-one (1.0 g, 5.0 mmol), cesium carbonate (1.6 g, 5.0 mmol), cuprous iodide ( 476.0 mg, 5.0 mmol) and N,N'-dimethylethylenediamine (440.0 mg, 5.0 mmol) were added to toluene (15 mL), replaced with nitrogen three times, and heated at 110 degrees Celsius under reflux for condensation and reacted overnight.
反应结束，加入乙酸乙酯(30毫升)，混合物用饱和食盐水(20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/9)。得到787.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：60.5％)。LCMS:RT＝3.16min,[M+H] +＝529.28。 After the reaction was completed, ethyl acetate (30 mL) was added, the mixture was washed with saturated brine (20 mL × 3 times), then the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography ( Eluent: methanol/dichloromethane=1/9). Obtained 787.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazine-1- (Yl)phenyl)tert-butyl propionate (yield: 60.5%). LCMS: RT = 3.16 min, [M+H] + =529.28.
步骤C：合成(S)-2-氨基-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step C: Synthesis of (S)-2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate
Figure PCTCN2020133493-appb-000077
Figure PCTCN2020133493-appb-000077
室温下，将(S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(300.0毫克，0.8毫摩尔)加入乙酸乙酯(8.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，2.0毫升)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate (300.0 mg, 0.8 mmol) was added to ethyl acetate (8.0 mL), and ethyl acetate solution of hydrogen chloride (2.0 mol/L, 2.0 mL) was added dropwise with stirring. After the addition, nitrogen React overnight at room temperature under protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到217.0毫克白色固体(S)-2-氨基-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝0.56min,[M+H] +＝432.28。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 217.0 mg of white solid ( S)-2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) tert-butyl propionate, the crude product is used directly in the next step reaction. LCMS: RT=0.56 min, [M+H] + =432.28.
步骤D：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯Step D: Synthesis of (S)-2-((Fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Tert-butyl propionate
Figure PCTCN2020133493-appb-000078
Figure PCTCN2020133493-appb-000078
室温下，将(S)-2-氨基-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯粗品和碳酸钠(297.0毫克，2.8毫摩尔)加入四氢呋喃(2.0毫升)和水(2.0毫升)的混合溶剂中，搅拌下加入芴甲氧羰酰氯(176.0毫克，0.7毫摩尔)，氮气保护下室温反应3.0小时。At room temperature, the crude product of (S)-2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid tert-butyl ester and Sodium carbonate (297.0 mg, 2.8 mmol) was added to a mixed solvent of tetrahydrofuran (2.0 ml) and water (2.0 ml), fluorenyl methoxycarbonyl chloride (176.0 mg, 0.7 mmol) was added with stirring, and the reaction was performed at room temperature under nitrogen for 3.0 hour.
反应结束，加入饱和氯化钠溶液(20毫升)，混合物用二氯甲烷(20毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩。所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/9)。得到240.0毫克白色固体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(收率：72.0％)。LCMS:RT＝3.39min,[M+H] +＝654.29。 After the reaction was completed, saturated sodium chloride solution (20 mL) was added, and the mixture was extracted with dichloromethane (20 mL×3 times). After the organic phases were combined, they were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtained 240.0 mg of white solid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)benzene Yl) tert-butyl propionate (yield: 72.0%). LCMS: RT = 3.39 min, [M+H] + =654.29.
步骤E：合成(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸Step E: Synthesis of (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Propionic acid
Figure PCTCN2020133493-appb-000079
Figure PCTCN2020133493-appb-000079
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸叔丁酯(240.0毫克，0.4毫摩尔)加入二氯甲烷(5.0毫升)中，搅拌下滴加三氟乙酸(2.5毫升)，滴加完毕后，氮气保护下室温反应1.0小时。At room temperature, (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Tert-Butyl propionate (240.0 mg, 0.4 mmol) was added to dichloromethane (5.0 mL), trifluoroacetic acid (2.5 mL) was added dropwise with stirring, and after the addition was completed, reacted at room temperature for 1.0 hour under nitrogen protection.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，得到黄色油状液体(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸，粗品直接用于下一步反应。LCMS:RT＝3.03min,[M+H] +＝598.27。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump to obtain a yellow oily liquid (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxy) Cyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionic acid, the crude product was directly used in the next reaction. LCMS: RT = 3.03 min, [M+H] + =598.27.
步骤F：合成(S)-4-(2-((芴甲氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯Step F: Synthesis of (S)-4-(2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000080
Figure PCTCN2020133493-appb-000080
室温下，将(S)-2-((芴甲氧羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酸粗品和4-氨基-1-叔丁氧羰基-吲哚-2-酸叔丁酯加入N,N-二甲基甲酰胺(2.0毫升)中，搅拌下滴加N,N-二异丙基乙胺，调节pH值至碱性。然后加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(285.0毫克，0.8毫摩尔)和N,N-二甲基甲酰胺(129.0毫克，1.0毫摩尔)，于氮气保护下室温反应过夜。At room temperature, (S)-2-((fluorenylmethoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Crude propionic acid and 4-amino-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester were added to N,N-dimethylformamide (2.0 ml), and N,N-dimethicone was added dropwise with stirring. Isopropylethylamine, adjust the pH to alkaline. Then add 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (285.0 mg, 0.8 mmol) and N,N-dimethylformaldehyde Amide (129.0 mg, 1.0 mmol) was reacted overnight at room temperature under nitrogen protection.
反应结束，加入乙酸乙酯(15毫升)，混合液用饱和食盐水(10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：甲醇/二氯甲烷＝1/10)。得到304.0毫克白色固体(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯。LCMS:RT＝3.90min,[M+H] +＝912.36。 After the reaction was completed, ethyl acetate (15 mL) was added, and the mixture was washed with 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 304.0 mg of white solid (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Propionamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester. LCMS: RT = 3.90 min, [M+H] + =912.36.
步骤G：合成(S)-4-(2-氨基-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step G: Synthesis of (S)-4-(2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido) -1H-Indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000081
Figure PCTCN2020133493-appb-000081
室温下，将(S)-4-(2-芴甲氧羰基氨基-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1-叔丁氧羰基-吲哚-2-酸叔丁酯(304.0毫克，0.3毫摩尔)加入二氯甲烷(6.0毫升)中，搅拌下滴加4-甲基哌啶(1.5毫升)，于氮气保护下室温反应1.0小时。At room temperature, (S)-4-(2-fluorenylmethoxycarbonylamino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl) Propanamido)-1-tert-butoxycarbonyl-indole-2-acid tert-butyl ester (304.0 mg, 0.3 mmol) was added to dichloromethane (6.0 ml), and 4-methylpiperidine ( 1.5 ml), and react at room temperature for 1.0 hour under the protection of nitrogen.
反应结束，直接减压浓缩。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153.0毫克白色固体(S)-4-(2-氨基-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯。LCMS:RT＝1.51min,[M+H] +＝624.23。 After the reaction was over, it was directly concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtain 153.0 mg of white solid (S)-4-(2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido ) -1H-Indole-2-acid tert-butyl ester. LCMS: RT=1.51 min, [M+H] + =624.23.
步骤H：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯Step H: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl ester
Figure PCTCN2020133493-appb-000082
Figure PCTCN2020133493-appb-000082
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(129.0毫克，0.4毫摩尔)、(S)-4-(2-氨基-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯(153.0毫克，0.4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(228.0毫克，0.6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(104.0毫克，0.8毫摩尔)，滴毕，N 2保护下，室温反应1.0小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (129.0 mg, 0.4 mmol), (S)-4-( 2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl Ester (153.0 mg, 0.4 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (228.0 mg, 0.6 mmol) were added To N,N-dimethylformamide (4.0 mL), N,N-diisopropylethylamine (104.0 mg, 0.8 mmol) was added dropwise, after dripping, the reaction was carried out at room temperature for 1.0 hour under N 2 protection.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得粗品直接用于下一步反应。LCMS:RT＝3.23min,[M+H] +＝839.26。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained crude product was directly used in the next reaction. LCMS: RT = 3.23 min, [M+H] + =839.26.
步骤I：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸Step I: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid
Figure PCTCN2020133493-appb-000083
Figure PCTCN2020133493-appb-000083
室温下，将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸叔丁酯粗品加入二氯甲烷(10.0毫升)中，滴加三氟乙酸(5.0毫升)，室温反应1.0小时。At room temperature, (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-acid tert-butyl crude product was added to two Trifluoroacetic acid (5.0 mL) was added dropwise to methyl chloride (10.0 mL), and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到93.0毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-酸。LCMS:RT＝2.87min,[M+H] +＝783.00。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is separated by a preparative liquid chromatograph to obtain 93.0 mg of white solid (S)-4-(2-(2-(((5-chloro -2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiper (Azin-1-yl)phenyl)propionamido)-1H-indole-2-acid. LCMS: RT = 2.87 min, [M+H] + =783.00.
实施例6Example 6
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(氧杂环丁烷-3-基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000084
Figure PCTCN2020133493-appb-000084
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(氧杂环丁烷-3-基)-2-氧代哌嗪Step A: Synthesis of 4-(oxetan-3-yl)-2-oxopiperazine
Figure PCTCN2020133493-appb-000085
Figure PCTCN2020133493-appb-000085
室温下，将2-氧代哌嗪(500毫克，5.0毫摩尔)和氧杂环丁-3-酮(720.0毫克，10.0毫摩尔)加入甲醇(20毫升)和乙酸(0.4毫升)的混合溶剂中，氮气保护下，室温搅拌1.0小时。然后在冰浴下，缓慢加入氰基硼氢化钠(692.0毫克，11.0毫摩尔)，升至室温后，反应过夜。At room temperature, add 2-oxopiperazine (500 mg, 5.0 mmol) and oxetan-3-one (720.0 mg, 10.0 mmol) to a mixed solvent of methanol (20 mL) and acetic acid (0.4 mL) Under the protection of nitrogen, stir at room temperature for 1.0 hour. Then in an ice bath, sodium cyanoborohydride (692.0 mg, 11.0 mmol) was slowly added, after warming to room temperature, the reaction was carried out overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用40毫升二氯甲烷洗涤，合并有机相，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351.0毫克白色固体4-(氧杂环丁烷-3-基)-2-氧代哌嗪(收率：45.0％)。After the reaction was completed, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 40 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtained 351.0 mg of white solid 4-(oxetan-3-yl)-2-oxopiperazine (yield: 45.0%).
步骤B：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(oxetan-3-yl)-2-oxopiperazine-1 -Yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000086
Figure PCTCN2020133493-appb-000086
氮气保护下，将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(582.0毫克，1.1毫摩尔)、4-(氧杂环丁烷-3-基)哌嗪-2-酮(351.0毫克，2.3毫摩尔)、碳酸铯(738.0毫克，2.3毫摩尔)、碘化亚铜(214.0毫克，1.1毫摩尔)和N,N’-二甲基乙二胺(198.0毫克，2.3毫摩尔)加入甲苯(10毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under nitrogen protection, (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (582.0 mg, 1.1 mmol) , 4-(oxetan-3-yl)piperazin-2-one (351.0 mg, 2.3 mmol), cesium carbonate (738.0 mg, 2.3 mmol), cuprous iodide (214.0 mg, 1.1 mmol) Mol) and N,N'-dimethylethylenediamine (198.0 mg, 2.3 mmol) were added to toluene (10 mL), replaced with nitrogen three times, condensed and refluxed and heated at 110 degrees Celsius to react overnight.
反应结束，加入乙酸乙酯(30毫升)，混合物用饱和食盐水20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/10)。得到167.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：25.0％)。LCMS:RT＝3.90min,[M-H] -＝617.24。 After the reaction was completed, ethyl acetate (30 ml) was added, the mixture was washed with saturated brine 20 ml × 3 times, and then the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue obtained was purified by silica gel column chromatography (washing). Removal agent: methanol/dichloromethane=1/10). Obtained 167.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(oxetan-3-yl)-2-oxopiperazine- 1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 25.0%). LCMS: RT = 3.90 min, [MH] - = 617.24.
步骤C：合成(S)-4-(2-氨基-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-amino-3-(4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamide Yl) tert-butyl benzoate
Figure PCTCN2020133493-appb-000087
Figure PCTCN2020133493-appb-000087
室温下，将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(167.0毫克，0.4毫摩尔)加入二氯甲烷(20.0毫升)中，搅拌下滴加三氟乙酸(912毫克)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(oxetan-3-yl)-2-oxopiperazine-1 -Yl)phenyl)propionamido)tert-butyl benzoate (167.0 mg, 0.4 mmol) was added to dichloromethane (20.0 ml), and trifluoroacetic acid (912 mg) was added dropwise with stirring. After the addition was complete, React overnight at room temperature under nitrogen protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到白色固体(S)-4-(2-氨基-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝2.77min,[M+H] +＝517.22。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (S) -4-(2-Amino-3-(4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate , The crude product is directly used in the next reaction. LCMS: RT = 2.77 min, [M+H] + =517.22.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000088
Figure PCTCN2020133493-appb-000088
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(64.0毫克，0.2毫摩尔)、(S)-4-(2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(95.0毫克，0.2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(114.0毫克，0.3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(52.0毫克，0.4毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (64.0 mg, 0.2 mmol), (S)-4-( Tert-Butyl 2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate (95.0 Mg, 0.2 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (114.0 mg, 0.3 mmol) add N,N -In dimethylformamide (5.0 ml), add N,N-diisopropylethylamine (52.0 mg, 0.4 mmol) dropwise, and after the dropwise , react under N 2 protection at room temperature for 5 hours.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10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：甲醇/二氯甲烷＝1/10)。得到78.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯。LCMS:RT＝3.90min,[M+H] +＝744.25。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with 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 78.0 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate. LCMS: RT = 3.90 min, [M+H] + =744.25.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000089
Figure PCTCN2020133493-appb-000089
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(78.0毫克，0.1毫摩尔)加入二氯甲烷(8.0毫升)中，滴加三氟乙酸(4.0毫升)，室温反应1.0小时。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(oxetan-3-yl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (78.0 mg, 0.1 mmol) was added Trifluoroacetic acid (4.0 mL) was added dropwise to dichloromethane (8.0 mL), and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到49.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(氧杂环丁烷-3-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯。LCMS:RT＝3.30min,[M+H] +＝688.17。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was separated by a preparative liquid chromatograph to obtain 49.0 mg of white solid (S)-4-(2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-(oxetan-3-yl)-2-oxo Piperazin-1-yl) phenyl) propionamido) tert-butyl benzoate. LCMS: RT = 3.30 min, [M+H] + =688.17.
实施例7Example 7
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(Tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000090
Figure PCTCN2020133493-appb-000090
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)-2-氧代哌嗪Step A: Synthesis of 4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine
Figure PCTCN2020133493-appb-000091
Figure PCTCN2020133493-appb-000091
室温下，将2-氧代哌嗪(500毫克，5.0毫摩尔)和四氢-2H-吡喃-4-酮(550.0毫克，5.5毫摩尔)加入甲醇(20毫升)和乙酸(0.4毫升)的混合溶剂中，氮气保护下，室温搅拌1.0小时。然后在冰浴下，缓慢加入氰基硼氢化钠(408.0毫克，6.5毫摩尔)，升至室温后，反应过夜。At room temperature, add 2-oxopiperazine (500 mg, 5.0 mmol) and tetrahydro-2H-pyran-4-one (550.0 mg, 5.5 mmol) to methanol (20 mL) and acetic acid (0.4 mL) In the mixed solvent, under the protection of nitrogen, stir at room temperature for 1.0 hour. Then in an ice bath, sodium cyanoborohydride (408.0 mg, 6.5 mmol) was slowly added, after warming to room temperature, the reaction was carried out overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用40毫升二氯甲烷洗涤，合并有机相，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552.4毫克白色固体4-(四氢-2H-吡喃-4-基)-2-氧代哌嗪(收率：54.5％)。After the reaction was completed, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 40 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). Obtained 552.4 mg of white solid 4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine (yield: 54.5%).
步骤B：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazine -1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000092
Figure PCTCN2020133493-appb-000092
氮气保护下，将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(779.0毫克，1.5毫摩尔)、4-(四氢-2H-吡喃-4-基)哌嗪-2-酮(552.4毫克，3.0毫摩尔)、碳酸铯(984.0毫克，3.0毫摩尔)、碘化亚铜(286.0毫克，1.5毫摩尔)和N,N’-二甲基乙二胺(265.0毫克，3.0毫摩尔)加入甲苯(10毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (779.0 mg, 1.5 mmol) , 4-(Tetrahydro-2H-pyran-4-yl)piperazin-2-one (552.4 mg, 3.0 mmol), cesium carbonate (984.0 mg, 3.0 mmol), cuprous iodide (286.0 mg, 1.5 mmol) and N,N'-dimethylethylenediamine (265.0 mg, 3.0 mmol) were added to toluene (10 mL), replaced with nitrogen three times, and heated at 110 degrees Celsius under reflux for condensation and reacted overnight.
反应结束，加入乙酸乙酯(30毫升)，混合物用饱和食盐水20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/5)。得到258.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：27.5％)。LCMS:RT＝3.32min,[M+H] +＝623.35。 After the reaction was completed, ethyl acetate (30 ml) was added, the mixture was washed with saturated brine 20 ml × 3 times, and then the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue obtained was purified by silica gel column chromatography (washing). Removal agent: ethyl acetate/n-hexane=1/5). Obtained 258.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxypiper (Azin-1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 27.5%). LCMS: RT = 3.32 min, [M+H] + =623.35.
步骤C：合成(S)-4-(2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl) (Propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000093
Figure PCTCN2020133493-appb-000093
室温下，将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(258.0毫克，0.4毫摩尔)加入乙酸乙酯(16.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，4.0毫升)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxypiperazine -1-yl) phenyl) propionamido) tert-butyl benzoate (258.0 mg, 0.4 mmol) was added to ethyl acetate (16.0 ml), and an ethyl acetate solution of hydrogen chloride (2.0 mol/L) was added dropwise with stirring. , 4.0 ml), after the addition is complete, react overnight at room temperature under nitrogen protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到白色固体(S)-4-(2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝1.70min,[M+H] +＝523.25。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (S) -4-(2-Amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido) benzoic acid tert Butyl ester, the crude product is directly used in the next reaction. LCMS: RT=1.70 min, [M+H] + =523.25.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000094
Figure PCTCN2020133493-appb-000094
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(107.0毫克，0.4毫摩尔)、(S)-4-(2-氨基-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(173.0毫克，0.3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(190.0毫克，0.5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(85.0毫克，0.7毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (107.0 mg, 0.4 mmol), (S)-4-( Tert-Butyl 2-amino-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate (173.0 Mg, 0.3 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (190.0 mg, 0.5 mmol) add N,N -To dimethylformamide (5.0 ml), add N,N-diisopropylethylamine (85.0 mg, 0.7 mmol) dropwise, after dropping , react under N 2 protection at room temperature for 5 hours.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10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：甲醇/二氯甲烷＝1/10)。得到145.6毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：56.8％)。LCMS:RT＝3.37min,[M+H] +＝772.31。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with 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). 145.6 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(Tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate (Yield: 56.8% ). LCMS: RT = 3.37 min, [M+H] + =772.31.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000095
Figure PCTCN2020133493-appb-000095
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(145.6毫克，0.2毫摩尔)加入二氯甲烷(8.0毫升)中，滴加三氟乙酸(4.0毫升)，室温反应1.0小时。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Tetrahydro-2H-pyran-4-yl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (145.6 mg, 0.2 mmol ) Was added to dichloromethane (8.0 ml), trifluoroacetic acid (4.0 ml) was added dropwise, and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到62.3毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(四氢-2H-吡喃-4-基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：52.9％)。LCMS:RT＝2.89min,[M+H] +＝716.18。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was separated by a preparative liquid chromatograph to obtain 62.3 mg of white solid (S)-4-(2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-(tetrahydro-2H-pyran-4-yl)-2 -Oxypiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 52.9%). LCMS: RT = 2.89 min, [M+H] + =716.18.
实施例8Example 8
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(2-oxo-4--(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000096
Figure PCTCN2020133493-appb-000096
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-羟基-2-氧代哌啶Step A: Synthesis of 4-hydroxy-2-oxopiperidine
Figure PCTCN2020133493-appb-000097
Figure PCTCN2020133493-appb-000097
冰浴下，将硼氢化钠(35.0毫克，0.9毫摩尔)加入含有2,4-二氧代哌啶(339.4毫克，3.0毫摩尔)乙醇(6毫升)中，冰浴下搅拌15分钟。Under ice bath, sodium borohydride (35.0 mg, 0.9 mmol) was added to ethanol (6 ml) containing 2,4-dioxopiperidine (339.4 mg, 3.0 mmol), and stirred for 15 minutes under ice bath.
反应结束，直接减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/正二氯甲烷＝1/9)。得到212.0毫克无色油状液体4-羟基-2-氧代哌啶(收率：61.4％)。After the reaction was completed, it was directly concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent: methanol/n-dichloromethane=1/9). Obtained 212.0 mg of colorless oily liquid 4-hydroxy-2-oxopiperidine (yield: 61.4%).
步骤B：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido )Tert-Butyl Benzoate
Figure PCTCN2020133493-appb-000098
Figure PCTCN2020133493-appb-000098
氮气保护下，将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(478.0毫克，0.9毫摩尔)、4-羟基-2-氧代哌啶(212.0毫克，1.8毫摩尔)、碳酸铯(604.0毫克，1.8毫摩尔)、碘化亚铜(176.0毫克，0.9毫摩尔)和N,N’-二甲基乙二胺(163.0毫克，1.8毫摩尔)加入甲苯(5毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (478.0 mg, 0.9 mmol) , 4-hydroxy-2-oxopiperidine (212.0 mg, 1.8 mmol), cesium carbonate (604.0 mg, 1.8 mmol), cuprous iodide (176.0 mg, 0.9 mmol) and N,N'-two Methylethylenediamine (163.0 mg, 1.8 mmol) was added to toluene (5 mL), replaced with nitrogen three times, and heated under reflux at 110 degrees Celsius for reaction overnight.
反应结束，加入乙酸乙酯(10毫升)，混合物用饱和食盐水10毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/8)。得到201.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：45.1％)。LCMS:RT＝3.86min,[M-H] -＝552.22。 After the reaction was completed, ethyl acetate (10 mL) was added, the mixture was washed with saturated brine 10 mL × 3 times, and then the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (washing). Removal agent: methanol/dichloromethane=1/8). Obtain 201.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamide Yl) tert-butyl benzoate (yield: 45.1%). LCMS: RT = 3.86 min, [MH] - =552.22.
步骤C：合成(S)-4-(2-氨基-3-(4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of tert-butyl (S)-4-(2-amino-3-(4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000099
Figure PCTCN2020133493-appb-000099
室温下，将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(201.0毫克，0.4毫摩尔)加入乙酸乙酯(16.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，4.0毫升)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido ) Tert-butyl benzoate (201.0 mg, 0.4 mmol) was added to ethyl acetate (16.0 ml), and the ethyl acetate solution of hydrogen chloride (2.0 mol/L, 4.0 ml) was added dropwise with stirring. After the addition, nitrogen React overnight at room temperature under protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到白色固体(S)-4-(2-氨基-3-(4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝2.77min,[M-H] -＝452.14。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (S) Tert-butyl -4-(2-amino-3-(4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)benzoate, the crude product was directly used in the next reaction. LCMS: RT = 2.77 min, [MH] - =452.14.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000100
Figure PCTCN2020133493-appb-000100
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(139.0毫克，0.3毫摩尔)、(S)-4-(2-氨基-3-(4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯粗品和2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(175.0毫克，0.5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(79.0毫克，0.6毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (139.0 mg, 0.3 mmol), (S)-4-( Crude tert-butyl 2-amino-3-(4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)benzoate and 2-(7-oxobenzotriazole )-N,N,N',N'-Tetramethylurea hexafluorophosphate (175.0mg, 0.5mmol) was added to N,N-dimethylformamide (5.0ml), and N,N- Diisopropylethylamine (79.0 mg, 0.6 mmol) was dripped and reacted overnight at room temperature under N 2 protection.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物通过进行硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝10/1)。得到135.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率62.8％)。LCMS:RT＝3.97min,[M+H] +＝703.26。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by performing silica gel column chromatography (eluent: dichloromethane/methanol=10/1). 135.0 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)benzoate (yield 62.8%). LCMS: RT = 3.97 min, [M+H] + =703.26.
步骤E：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000101
Figure PCTCN2020133493-appb-000101
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(129.0毫克，0.1毫摩尔)加入二氯甲烷(4.0毫升)中，滴加三氟乙酸(2毫升)，室温反应1.0小时。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4-(4-hydroxy-2-oxopiperidin-1-yl)phenyl)propionamido) tert-butyl benzoate (129.0 mg, 0.1 mmol) was added to two Trifluoroacetic acid (2 mL) was added dropwise to methyl chloride (4.0 mL), and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到40.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4-(4-羟基-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸。LCMS:RT＝3.30min,[M+H] +＝647.10。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was separated by a preparative liquid chromatograph to obtain 40.0 mg of white solid (S)-4-(2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(2-oxo-4-(4-hydroxy-2-oxopiper) (Pyridin-1-yl)phenyl)propionamido)benzoic acid. LCMS: RT = 3.30 min, [M+H] + =647.10.
实施例9Example 9
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(2-oxo-4--(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000102
Figure PCTCN2020133493-appb-000102
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxo-8-oxa-2-azaspiro[4.5]decane- 2-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000103
Figure PCTCN2020133493-appb-000103
氮气保护下，将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(389.0毫克，0.8毫摩尔)、3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷(232.8毫克，1.5毫摩尔)、碳酸铯(488.5毫克，1.5毫摩尔)、碘化亚铜(143.0毫克，0.8毫摩尔)和N,N’-二甲基乙二胺(135.0毫克，1.5毫摩尔)加入甲苯(5毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under nitrogen protection, (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (389.0 mg, 0.8 mmol) , 3-oxo-8-oxa-2-azaspiro[4.5]decane (232.8 mg, 1.5 mmol), cesium carbonate (488.5 mg, 1.5 mmol), cuprous iodide (143.0 mg, 0.8 Millimoles) and N,N'-dimethylethylenediamine (135.0 mg, 1.5 millimoles) were added to toluene (5 ml), replaced with nitrogen three times, and heated under reflux at 110 degrees Celsius for condensing and reacting overnight.
反应结束，加入乙酸乙酯(10毫升)，混合物用饱和食盐水10毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝2/1)。得到453.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：76.3％)。LCMS:RT＝4.16min,[M-H] -＝593.35。 After the reaction was completed, ethyl acetate (10 mL) was added, the mixture was washed with saturated brine 10 mL × 3 times, and then the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (washing). Removal agent: ethyl acetate/n-hexane=2/1). Obtained 453.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxo-8-oxa-2-azaspiro[4.5]decane -2-yl)phenyl)propionamido)tert-butyl benzoate (yield: 76.3%). LCMS: RT = 4.16 min, [MH] - =593.35.
步骤B：合成(S)-4-(2-氨基-3-(4-(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-amino-3-(4-(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propane Amido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000104
Figure PCTCN2020133493-appb-000104
室温下，将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(453.0毫克，0.8毫摩尔)加入乙酸乙酯(16.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，4.0毫升)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxo-8-oxa-2-azaspiro[4.5]decane- 2-yl)phenyl)propionamido)tert-butyl benzoate (453.0 mg, 0.8 mmol) was added to ethyl acetate (16.0 ml), and an ethyl acetate solution of hydrogen chloride (2.0 mol/L, 4.0 ml), after the addition is complete, react overnight at room temperature under nitrogen protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到白色固体(S)-4-(2-氨基-3-(4-(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝2.90min,[M] +＝493.17。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (S) -4-(2-Amino-3-(4-(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)tert-butyl benzoate Ester, the crude product is directly used in the next reaction. LCMS: RT = 2.90 min, [M] + =493.17.
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4-(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4-(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000105
Figure PCTCN2020133493-appb-000105
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(114.0毫克，0.2毫摩尔)、(S)-4-(2-氨基-3-(4-(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯粗品和2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(122.0毫克，0.3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(56.0毫克，0.4毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (114.0 mg, 0.2 mmol), (S)-4-( 2-amino-3-(4-(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido) tert-butyl benzoate crude and 2 -(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (122.0 mg, 0.3 mmol) was added to N,N-dimethylformamide (5.0 (Ml), N,N-diisopropylethylamine (56.0 mg, 0.4 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，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后 减压浓缩。所得残余物通过进行硅胶柱层析纯化(洗脱剂：二氯甲烷/甲醇＝30/1)。得到118.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯。LCMS:RT＝4.14min,[M-H] -＝741.15。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by performing silica gel column chromatography (eluent: dichloromethane/methanol=30/1). 118.0 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(2-oxo-4--(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)tert-butyl benzoate Ester. LCMS: RT = 4.14 min, [MH] - =741.15.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000106
Figure PCTCN2020133493-appb-000106
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4-(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(118.0毫克，0.1毫摩尔)加入二氯甲烷(8.0毫升)中，滴加三氟乙酸(4毫升)，室温反应1.0小时。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4-(3-oxo-8-oxa-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)tert-butyl benzoate (118.0 mg, 0.1 mmol) was added to dichloromethane (8.0 mL), trifluoroacetic acid (4 mL) was added dropwise, and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到40.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4-(3-氧代-8-氧杂-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸。LCMS:RT＝4.14min,[M-H] -＝741.15。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was separated by a preparative liquid chromatograph to obtain 40.0 mg of white solid (S)-4-(2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(2-oxo-4-(3-oxo-8-oxa -2-Azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid. LCMS: RT = 4.14 min, [MH] - =741.15.
实施例10Example 10
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2,2,2-三氟乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000107
Figure PCTCN2020133493-appb-000107
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(2,2,2-三氟乙基)哌嗪-2-酮Step A: Synthesis of 4-(2,2,2-trifluoroethyl)piperazin-2-one
Figure PCTCN2020133493-appb-000108
Figure PCTCN2020133493-appb-000108
室温下，将2-氧代哌嗪(3.0克，30.0毫摩尔)、2,2,2-三氟乙基三氟甲磺酸酯(6.5克，28.0毫摩尔)和碳酸铯(15.0克，46.2毫摩尔)加入乙腈(40毫升)中，氮气保护下，冷凝回流，加热80摄氏度过夜。At room temperature, 2-oxopiperazine (3.0 g, 30.0 mmol), 2,2,2-trifluoroethyl triflate (6.5 g, 28.0 mmol) and cesium carbonate (15.0 g, 46.2 mmol) was added to acetonitrile (40 ml), under the protection of nitrogen, condensed to reflux, and heated at 80 degrees Celsius overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用40毫升二氯甲烷洗涤，合并有机相，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2.4克白色固体4-(2,2,2-三氟乙基)哌嗪-2-酮(收率：47.1％)。After the reaction was completed, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 40 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtained 2.4 g of white solid 4-(2,2,2-trifluoroethyl)piperazin-2-one (yield: 47.1%).
步骤B：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazine- 1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000109
Figure PCTCN2020133493-appb-000109
氮气保护下，将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(779.0毫克，1.5毫摩尔)、4-(2,2,2-三氟乙基)哌嗪-2-酮(546.4毫克，3.0毫摩尔)、碳酸铯(977.0毫克，3.0毫摩尔)、碘化亚铜(285.0毫克，1.5毫摩尔) 和N,N’-二甲基乙二胺(270.0毫克，3.0毫摩尔)加入甲苯(10毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (779.0 mg, 1.5 mmol) , 4-(2,2,2-Trifluoroethyl)piperazin-2-one (546.4 mg, 3.0 mmol), cesium carbonate (977.0 mg, 3.0 mmol), cuprous iodide (285.0 mg, 1.5 MM) and N,N'-dimethylethylenediamine (270.0 mg, 3.0 mmol) were added to toluene (10 mL), replaced with nitrogen three times, and heated at 110°C under reflux for condensation and reacted overnight.
反应结束，加入乙酸乙酯(30毫升)，混合物用饱和食盐水20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/30)。得到742.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：79.8％)。LCMS:RT＝4.25min,[M-H] -＝619.23。 After the reaction was completed, ethyl acetate (30 ml) was added, the mixture was washed with saturated brine 20 ml × 3 times, and then the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue obtained was purified by silica gel column chromatography (washing). Removal agent: methanol/dichloromethane=1/30). Obtain 742.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazine -1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 79.8%). LCMS: RT = 4.25 min, [MH] - =619.23.
步骤C：合成(S)-4-(2-氨基-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-amino-3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propane Amido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000110
Figure PCTCN2020133493-appb-000110
室温下，将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(742.0毫克，1.2毫摩尔)加入乙酸乙酯(16.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，4.0毫升)，滴加完毕后，氮气保护下室温反应4.0小时。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazine- 1-yl)phenyl)propionamido)tert-butyl benzoate (742.0 mg, 1.2 mmol) was added to ethyl acetate (16.0 ml), and an ethyl acetate solution of hydrogen chloride (2.0 mol/L, 4.0 ml), after the addition is complete, react at room temperature for 4.0 hours under nitrogen protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到白色固体(S)-4-(2-氨基-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝3.04min,[M] +＝520.80。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (S) -4-(2-amino-3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate Ester, the crude product is directly used in the next reaction. LCMS: RT = 3.04 min, [M] + =520.80.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000111
Figure PCTCN2020133493-appb-000111
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(320.0毫克，1.2毫摩尔)、(S)-4-(2-氨基-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(515.0毫克，1.0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(570.0毫克，1.5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(258.0毫克，2.0毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (320.0 mg, 1.2 mmol), (S)-4-( Tert-Butyl 2-amino-3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate (515.0 mg , 1.0 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (570.0 mg, 1.5 mmol) was added to N,N- In dimethylformamide (5.0 ml), N,N-diisopropylethylamine (258.0 mg, 2.0 mmol) was added dropwise, and after the dropping, the reaction was carried out at room temperature for 5 hours under the protection of N 2.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10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：甲醇/二氯甲烷＝1/9)。得到264.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：34.3％)。LCMS:RT＝4.23min,[M-H] -＝770.34。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with 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=1/9). Obtained 264.0 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (Yield: 34.3%) . LCMS: RT = 4.23 min, [MH] - =770.34.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2,2,2-trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000112
Figure PCTCN2020133493-appb-000112
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(51.0毫克，0.06毫摩尔)加入二氯甲烷(4.0毫升)中，滴加三氟乙酸(2毫升)，室温反应1.0小时。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(2,2,2-Trifluoroethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (51.0 mg, 0.06 mmol) Add dichloromethane (4.0 mL), add trifluoroacetic acid (2 mL) dropwise, and react at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到19.8毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2,2,2-三氟乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：56.5％)。LCMS:RT＝3.72min,[M-H] -＝712.18。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was separated by a preparative liquid chromatograph to obtain 19.8 mg of white solid (S)-4-(2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-(2,2,2-trifluoroethyl)-2- Oxypiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 56.5%). LCMS: RT = 3.72 min, [MH] - =712.18.
实施例11Example 11
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(2-oxo-4--(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000113
Figure PCTCN2020133493-appb-000113
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxo-2-azaspiro[4.5]decane-2-yl)benzene (Yl) propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000114
Figure PCTCN2020133493-appb-000114
氮气保护下，将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(779.0毫克，1.5毫摩尔)、3-氧代-2-氮杂螺[4.5]癸烷(459.0毫克，3.0毫摩尔)、碳酸铯(975.0毫克，3.0毫摩尔)、碘化亚铜(286.0毫克，1.5毫摩尔)和N,N’-二甲基乙二胺(270.0毫克，3.0毫摩尔)加入甲苯(5毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (779.0 mg, 1.5 mmol) , 3-oxo-2-azaspiro[4.5]decane (459.0 mg, 3.0 mmol), cesium carbonate (975.0 mg, 3.0 mmol), cuprous iodide (286.0 mg, 1.5 mmol) and N ,N'-Dimethylethylenediamine (270.0 mg, 3.0 mmol) was added to toluene (5 mL), replaced with nitrogen three times, and heated at 110°C under reflux for condensation and reacted overnight.
反应结束，加入乙酸乙酯(10毫升)，混合物用饱和食盐水10毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝1/1)。得到598.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：67.4％)。LCMS:RT＝4.76min,[M-H] -＝590.25。 After the reaction was completed, ethyl acetate (10 ml) was added, the mixture was washed with saturated brine 10 ml × 3 times, then the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (washing). Removal agent: ethyl acetate/n-hexane=1/1). Obtain 598.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxo-2-azaspiro[4.5]decane-2-yl) (Phenyl) propionamido) tert-butyl benzoate (yield: 67.4%). LCMS: RT = 4.76 min, [MH] - =590.25.
步骤B：合成(S)-4-(2-氨基-3-(4-(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-amino-3-(4-(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid Tert-butyl ester
Figure PCTCN2020133493-appb-000115
Figure PCTCN2020133493-appb-000115
室温下，将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(598.0毫克，1.1毫摩尔)加入乙酸乙酯(16.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，4.0毫升)，滴加完毕后，氮气保护下室温反应过夜。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxo-2-azaspiro[4.5]decane-2-yl)benzene (L) propionamido) tert-butyl benzoate (598.0 mg, 1.1 mmol) was added to ethyl acetate (16.0 mL), and ethyl acetate solution of hydrogen chloride (2.0 mol/L, 4.0 mL) was added dropwise with stirring. After the addition is complete, react overnight at room temperature under nitrogen protection.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到白色固体(S)-4-(2-氨基-3-(4-(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝3.22min,[M-H] -＝490.21。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (S) -4-(2-Amino-3-(4-(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido) tert-butyl benzoate, use the crude product directly In the next step. LCMS: RT = 3.22 min, [MH] - = 490.21.
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4-(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4-(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000116
Figure PCTCN2020133493-appb-000116
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(375.0毫克，1.4毫摩尔)、(S)-4-(2-氨基-3-(4-(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯粗品和2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(685.0毫克，1.8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(300.0毫克，2.4毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (375.0 mg, 1.4 mmol), (S)-4-( 2-amino-3-(4-(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido) tert-butyl benzoate crude and 2-(7-oxidation Benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (685.0mg, 1.8mmol) was added to N,N-dimethylformamide (5.0ml) and dropped Add N,N-diisopropylethylamine (300.0 mg, 2.4 mmol), drip it, and react under N 2 protection at room temperature overnight.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物通过进行硅胶柱层析纯化(洗脱剂：乙酸乙酯/正己烷＝3/5)。得到455.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯。LCMS:RT＝4.62min,[M+H] +＝763.29。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by performing silica gel column chromatography (eluent: ethyl acetate/n-hexane=3/5). Obtained 455.0 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(2-oxo-4–(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoate. LCMS: RT = 4.62 min, [M+H] + =763.29.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000117
Figure PCTCN2020133493-appb-000117
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4-(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(100毫克，0.1毫摩尔)加入二氯甲烷(8.0毫升)中，滴加三氟乙酸(4毫升)，室温反应1.0小时。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4-(3-oxo-2-azaspiro[4.5]decane-2-yl)phenyl)propionamido) tert-butyl benzoate (100 mg, 0.1 (Mmol) was added to dichloromethane (8.0 mL), trifluoroacetic acid (4 mL) was added dropwise, and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到68.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4-(3-氧代-2-氮杂螺[4.5]癸烷-2-基)苯基)丙酰胺基)苯甲酸。LCMS:RT＝4.06min,[M-H] -＝707.15。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is separated by a preparative liquid chromatograph to obtain 68.0 mg of white solid (S)-4-(2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(2-oxo-4-(3-oxo-2-aza Spiro[4.5]decane-2-yl)phenyl)propionamido)benzoic acid. LCMS: RT = 4.06 min, [MH] - =707.15.
实施例12Example 12
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(2-oxo-4--(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000118
Figure PCTCN2020133493-appb-000118
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)甲基)哌嗪-2-酮Step A: Synthesis of 4-((tetrahydro-2H-pyran-4-yl)methyl)piperazin-2-one
Figure PCTCN2020133493-appb-000119
Figure PCTCN2020133493-appb-000119
室温下，将2-氧代哌嗪(500.0毫克，5.0毫摩尔)、4-(2-溴乙基)四氢-2H-吡喃(1.0克，5.5毫摩尔)和碳酸钾(1.8克，12.5毫摩尔)加入乙腈(15毫升)中，氮气保护下，冷凝回流，加热80摄氏度过夜。At room temperature, 2-oxopiperazine (500.0 mg, 5.0 mmol), 4-(2-bromoethyl)tetrahydro-2H-pyran (1.0 g, 5.5 mmol) and potassium carbonate (1.8 g, 12.5 mmol) was added to acetonitrile (15 ml), under nitrogen protection, condensed to reflux, and heated at 80 degrees Celsius overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用20毫升二氯甲烷洗涤，合并有机相，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444.0毫克白色固体4-((四氢-2H-吡喃-4-基)甲基)哌嗪-2-酮(收率：44.8％)。At the end of the reaction, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 20 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). Obtained 444.0 mg of white solid 4-((tetrahydro-2H-pyran-4-yl)methyl)piperazin-2-one (yield: 44.8%).
步骤B：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(((tetrahydro-2H-pyran-4-yl)methyl)piperazine-1 -Yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000120
Figure PCTCN2020133493-appb-000120
氮气保护下，将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(571.0毫克，1.1毫摩尔)、4-((四氢-2H-吡喃-4-基)甲基)哌嗪-2-酮(444.0毫克，2.2毫摩尔)、碳酸铯(717.0毫克，2.2毫摩尔)、碘化亚铜(209.5毫克，1.1毫摩尔)和N,N’-二甲基乙二胺(194.0毫克，2.2毫摩尔)加入甲苯(4毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under the protection of nitrogen, (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (571.0 mg, 1.1 mmol) , 4-((Tetrahydro-2H-pyran-4-yl)methyl)piperazin-2-one (444.0 mg, 2.2 mmol), cesium carbonate (717.0 mg, 2.2 mmol), cuprous iodide (209.5 mg, 1.1 mmol) and N,N'-dimethylethylenediamine (194.0 mg, 2.2 mmol) were added to toluene (4 mL), replaced with nitrogen three times, condensed and refluxed and heated at 110 degrees Celsius to react overnight.
反应结束，加入乙酸乙酯(10毫升)，混合物用饱和食盐水10毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/9)。得到583.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：83.3％)。LCMS:RT＝3.46min,[M-H] -＝636.35。 After the reaction was completed, ethyl acetate (10 ml) was added, the mixture was washed with saturated brine 10 ml × 3 times, then the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (washing). Removal agent: methanol/dichloromethane=1/9). Obtained 583.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(((tetrahydro-2H-pyran-4-yl)methyl)piperazine- 1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 83.3%). LCMS: RT = 3.46 min, [MH] - =636.35.
步骤C：合成(S)-4-(2-氨基-3-(4-(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-amino-3-(4-(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamide Yl) tert-butyl benzoate
Figure PCTCN2020133493-appb-000121
Figure PCTCN2020133493-appb-000121
室温下，将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(566.0毫克，1.1毫摩尔)加入乙酸乙酯(8.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，2.0毫升)，滴加完毕后，氮气保护下室温反应3.0小时。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(((tetrahydro-2H-pyran-4-yl)methyl)piperazine-1 -Yl)phenyl)propionamido)tert-butyl benzoate (566.0 mg, 1.1 mmol) was added to ethyl acetate (8.0 ml), and an ethyl acetate solution of hydrogen chloride (2.0 mol/L, 2.0 Ml), after the addition is complete, react at room temperature for 3.0 hours under the protection of nitrogen.
反应结束，加入饱和碳酸氢钠溶液(30毫升)，混合物用二氯甲烷(30毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到白色固体(S)-4-(2-氨基-3-(4-(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝2.13min,[M+H] +＝538.24。 After the reaction was completed, saturated sodium bicarbonate solution (30 mL) was added, the mixture was extracted with dichloromethane (30 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (S) -4-(2-amino-3-(4-(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamido) tert-butyl benzoate , The crude product is directly used in the next reaction. LCMS: RT = 2.13 min, [M+H] + =538.24.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000122
Figure PCTCN2020133493-appb-000122
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(370.0毫克，1.4毫摩尔)、(S)-4-(2-氨基-3-(4-(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯粗品和2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(570.0毫克，1.5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(260.0毫克，2.0毫摩尔)，滴毕，N 2保护下，室温反应3小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (370.0 mg, 1.4 mmol), (S)-4-( 2-amino-3-(4-(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamido) tert-butyl benzoate crude and 2- (7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (570.0 mg, 1.5 mmol) was added to N,N-dimethylformamide (5.0 ml ), N,N-diisopropylethylamine (260.0 mg, 2.0 mmol) was added dropwise, and after the dropping, the reaction was carried out at room temperature for 3 hours under the protection of N 2.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物通过进行硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/20)。得到75.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：9.5％)。LCMS:RT＝3.48min,[M+H] +＝786.23。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The obtained residue was purified by performing silica gel column chromatography (eluent: methanol/dichloromethane = 1/20). Obtain 75.0 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(2-oxo-4--(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (Yield: 9.5%). LCMS: RT = 3.48 min, [M+H] + =786.23.
步骤E：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000123
Figure PCTCN2020133493-appb-000123
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(75.0毫克，0.09毫摩尔)加入二氯甲烷(8.0毫升)中，滴加三氟乙酸(4毫升)，室温反应1.0小时。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxo-4--(((tetrahydro-2H-pyran-4-yl)methyl)piperazin-1-yl)phenyl)propionamido) tert-butyl benzoate( 75.0 mg, 0.09 mmol) was added to dichloromethane (8.0 mL), trifluoroacetic acid (4 mL) was added dropwise, and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到24.6毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代-4–(((四氢-2H-吡喃-4-基)甲基)哌嗪-1-基)苯基)丙酰胺基)苯甲酸。LCMS:RT＝2.93min,[M-H] -＝728.15。 After the reaction, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The residue obtained was separated by a preparative liquid chromatograph to obtain 24.6 mg of white solid (S)-4-(2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(2-oxo-4--(((tetrahydro-2H-pyran) -4-yl)methyl)piperazin-1-yl)phenyl)propionamido)benzoic acid. LCMS: RT = 2.93 min, [MH] - = 728.15.
实施例13Example 13
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000124
Figure PCTCN2020133493-appb-000124
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 PCTCN2020133493-appb-000125
Figure PCTCN2020133493-appb-000125
室温下，将2-氧代哌嗪(500.0毫克，5.0毫摩尔)、2-甲氧基-1-溴乙烷(0.7克，5.0毫摩尔)和碳酸钾(1.7克，12.5毫摩尔)加入乙腈(20毫升)中，氮气保护下，冷凝回流，加热80摄氏度过夜。At room temperature, add 2-oxopiperazine (500.0 mg, 5.0 mmol), 2-methoxy-1-bromoethane (0.7 g, 5.0 mmol) and potassium carbonate (1.7 g, 12.5 mmol) Condensate to reflux in acetonitrile (20 ml) under nitrogen protection, and heat at 80°C overnight.
反应结束，垫无水硫酸镁抽滤，滤饼用20毫升二氯甲烷洗涤，合并有机相，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765.0毫克白色固体4-(2-甲氧基乙基)哌嗪-2-酮(收率：97.0％)。At the end of the reaction, a pad of anhydrous magnesium sulfate was suction filtered, the filter cake was washed with 20 ml of dichloromethane, the organic phases were combined, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane=1/9). 765.0 mg of white solid 4-(2-methoxyethyl)piperazin-2-one was obtained (yield: 97.0%).
步骤B：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000126
Figure PCTCN2020133493-appb-000126
氮气保护下，将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(777.0毫克，1.5毫摩尔)、4-(2-甲氧基乙基)哌嗪-2-酮(465.0毫克，2.9毫摩尔)、碳酸铯(945.0毫克，2.9毫摩尔)、碘化亚铜(285.0毫克，1.5毫摩尔)和N,N’-二甲基乙二胺(255.6毫克，2.9毫摩尔)加入甲苯(10毫升)中，氮气置换三次，冷凝回流下110摄氏度加热反应过夜。Under nitrogen protection, (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (777.0 mg, 1.5 mmol) , 4-(2-Methoxyethyl)piperazin-2-one (465.0 mg, 2.9 mmol), cesium carbonate (945.0 mg, 2.9 mmol), cuprous iodide (285.0 mg, 1.5 mmol) And N,N'-dimethylethylenediamine (255.6 mg, 2.9 mmol) were added to toluene (10 mL), replaced with nitrogen three times, and heated under reflux at 110 degrees Celsius for reaction overnight.
反应结束，加入乙酸乙酯(30毫升)，混合物用饱和食盐水20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，减压浓缩，所得残余物用硅胶柱层析纯化(洗脱剂：甲醇/二氯甲烷＝1/20)。得到395.0毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：44.0％)。LCMS:RT＝1.98min,[M-H] -＝597.34。 After the reaction was completed, ethyl acetate (30 ml) was added, the mixture was washed with saturated brine 20 ml × 3 times, and then the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue obtained was purified by silica gel column chromatography (washing). Removal agent: methanol/dichloromethane=1/20). Obtain 395.0 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2-methoxyethyl)-2-oxopiperazine-1- (Yl)phenyl)propionamido)tert-butyl benzoate (yield: 44.0%). LCMS: RT=1.98 min, [MH] - =597.34.
步骤C：合成(S)-4-(2-氨基-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-amino-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) Tert-butyl benzoate
Figure PCTCN2020133493-appb-000127
Figure PCTCN2020133493-appb-000127
室温下，将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(200.0毫克，0.3毫摩尔)加入乙酸乙酯(4.0毫升)中，搅拌下滴加氯化氢的乙酸乙酯溶液(2.0摩尔/升，1.0毫升)，滴加完毕后，氮气保护下室温反应4.0小时。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)tert-butyl benzoate (200.0 mg, 0.3 mmol) was added to ethyl acetate (4.0 mL), and the ethyl acetate solution of hydrogen chloride (2.0 mol/L, 1.0 mL) was added dropwise with stirring. After the addition is complete, react at room temperature for 4.0 hours under nitrogen protection.
反应结束，加入饱和碳酸氢钠溶液(15毫升)，混合物用二氯甲烷(20毫升×3次)萃取，有机相合并后，用无水硫酸钠干燥，减压浓缩，得到白色固体(S)-4-(2-氨基-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯，粗品直接用于下一步反应。LCMS:RT＝1.75min,[M+H] +＝497.24。 After the reaction was completed, saturated sodium bicarbonate solution (15 mL) was added, the mixture was extracted with dichloromethane (20 mL×3 times), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (S) Tert-Butyl -4-(2-amino-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate, crude Used directly in the next reaction. LCMS: RT=1.75 min, [M+H] + =497.24.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000128
Figure PCTCN2020133493-appb-000128
室温下，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(175.0毫克，0.7毫摩尔)、(S)-4-(2-氨基-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(100.0毫克，0.6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(342.0毫克，0.9毫摩尔)加入N,N-二甲基甲酰胺(2.0毫升)中，滴加N,N-二异丙基乙胺(155.0毫克，1.2毫摩尔)，滴毕，N 2保护下，室温反应3小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (175.0 mg, 0.7 mmol), (S)-4-( Tert-butyl 2-amino-3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate (100.0 mg, 0.6 mmol Mol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (342.0 mg, 0.9 mmol) was added to N,N-dimethyl To formamide (2.0 ml), N,N-diisopropylethylamine (155.0 mg, 1.2 mmol) was added dropwise, and after the dropping was completed, the reaction was carried out at room temperature for 3 hours under the protection of N 2.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10毫升×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩。所得残余物通过制备液相色谱仪分离。得到35.0毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：23.3％)。LCMS:RT＝3.39min,[M+H] +＝746.27。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with saturated brine (10 mL×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was separated by preparative liquid chromatograph. Obtained 35.0 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate (yield: 23.3%). LCMS: RT = 3.39 min, [M+H] + =746.27.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000129
Figure PCTCN2020133493-appb-000129
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(35.0毫克，0.05毫摩尔)加入二氯甲烷(3.0毫升)中，滴加三氟乙酸(1.5毫升)，室温反应1.0小时。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(2-Methoxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (35.0 mg, 0.05 mmol) and dichloro Trifluoroacetic acid (1.5 mL) was added dropwise to methane (3.0 mL), and the reaction was carried out at room temperature for 1.0 hour.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到18.3毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：56.5％)。LCMS:RT＝2.91min,[M-H] -＝690.19。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is separated by a preparative liquid chromatograph to obtain 18.3 mg of white solid (S)-4-(2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-(2-methoxyethyl)-2-oxopiperazine -1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 56.5%). LCMS: RT = 2.91 min, [MH] - = 690.19.
实施例14Example 14
合成(S)-4-(2-(2-(((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5,6-Dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000130
Figure PCTCN2020133493-appb-000130
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5,6-二氯-2-(1H-四唑-1-基)硝基苯Step A: Synthesis of 5,6-dichloro-2-(1H-tetrazol-1-yl)nitrobenzene
Figure PCTCN2020133493-appb-000131
Figure PCTCN2020133493-appb-000131
室温下，将5,6-二氯-2-氟硝基苯(1.0克，4.8毫摩尔)、四氮唑(407.0毫克，5.8毫摩尔)和碳酸钾(993.6毫克，7.2毫摩尔)加入N,N-二甲基甲酰胺(10.0毫升)中，N 2保护下，80摄氏度反应1.5小时。 At room temperature, 5,6-dichloro-2-fluoronitrobenzene (1.0 g, 4.8 mmol), tetrazolium (407.0 mg, 5.8 mmol) and potassium carbonate (993.6 mg, 7.2 mmol) were added to N , N-dimethylformamide (10.0 ml), under N 2 protection, react at 80 degrees Celsius for 1.5 hours.
反应结束，加入乙酸乙酯30毫升，有机相混合物用饱和食盐水(20毫升×3次)洗涤，然后用无水硫酸钠干燥。有机相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539.0毫克白色固体5,6-二氯-2-(1H-四唑-1-基)硝基苯(收率：44.9％)。After the reaction was completed, 30 mL of ethyl acetate was added, and the organic phase mixture was washed with saturated brine (20 mL×3 times), and then dried over anhydrous sodium sulfate. The residue obtained by concentrating the organic phase under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=1/5). Obtained 539.0 mg of white solid 5,6-dichloro-2-(1H-tetrazol-1-yl)nitrobenzene (yield: 44.9%).
步骤B：合成5,6-二氯-2-(1H-四唑-1-基)苯胺Step B: Synthesis of 5,6-dichloro-2-(1H-tetrazol-1-yl)aniline
Figure PCTCN2020133493-appb-000132
Figure PCTCN2020133493-appb-000132
室温下，将5,6-二氯-2-(1H-四唑-1-基)硝基苯(539.0毫克，2.1毫摩尔)加入乙酸乙酯(20毫升)中，冰浴下，分批加入氯化亚锡二水合物(4.6克，20.4毫摩尔)，N 2保护下，室温反应过夜。 At room temperature, 5,6-dichloro-2-(1H-tetrazol-1-yl)nitrobenzene (539.0 mg, 2.1 mmol) was added to ethyl acetate (20 mL), under ice bath, in batches Add stannous chloride dihydrate (4.6 g, 20.4 mmol), and react at room temperature overnight under N 2 protection.
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30毫升乙酸乙酯洗涤，滤液用乙酸乙酯(30毫升×3次)萃取，合并有机相，然后用无水硫酸钠干燥，最后减压浓缩。得到376.0毫克黄色固体5,6-二氯-2-(1H-四唑-1-基)苯胺，直接用于下一步反应。LCMS:RT＝3.52min,[M+H] +＝230.07。 After the reaction was completed, it was quenched with saturated sodium bicarbonate aqueous solution, and excess sodium bicarbonate solid was added to adjust the pH to weakly alkaline. The filter cake was filtered with celite and the filter cake was washed with 30 ml of ethyl acetate. The filtrate was washed with ethyl acetate ( (30 ml × 3 times) extraction, combined organic phases, then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtained 376.0 mg of yellow solid 5,6-dichloro-2-(1H-tetrazol-1-yl)aniline, which was directly used in the next reaction. LCMS: RT = 3.52 min, [M+H] + = 230.07.
步骤C：合成2-((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸甲酯Step C: Synthesis of methyl 2-((5,6-dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000133
Figure PCTCN2020133493-appb-000133
N 2保护、冰浴下，向含有5,6-二氯-2-(1H-四唑-1-基)苯胺(100.0毫克，0.4毫摩尔)和吡啶(70.0毫克，0.9毫摩尔)的二氯甲烷(10毫升)中滴加草酰氯单甲酯(65.0毫克，0.5毫摩尔)，滴毕，室温反应30分钟。 Under the protection of N 2 and ice bath, add 5,6-dichloro-2-(1H-tetrazol-1-yl)aniline (100.0 mg, 0.4 mmol) and pyridine (70.0 mg, 0.9 mmol) to two Add oxalyl chloride monomethyl ester (65.0 mg, 0.5 mmol) dropwise to methyl chloride (10 ml), and then react at room temperature for 30 minutes.
反应结束，加入二氯甲烷(10毫升)，有机相先用饱和食盐水(15毫升×3次)洗涤，然后用无水硫酸钠干燥。有机相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320毫克白色固体2-((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸甲酯(收率：48.9％)。LCMS:RT＝3.21min,[M-H] -＝314.02。 After the reaction was completed, dichloromethane (10 mL) was added, and the organic phase was washed with saturated brine (15 mL×3 times) and then dried with anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure and the residue obtained was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/3) to obtain 320 mg of white solid 2-((5,6-dichloro-2-( 1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate methyl ester (yield: 48.9%). LCMS: RT = 3.21 min, [MH] - =314.02.
步骤D：合成2-((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸Step D: Synthesis of 2-((5,6-Dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000134
Figure PCTCN2020133493-appb-000134
冰浴下，向含有2-((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸甲酯(150.0毫克，0.5毫摩尔)的四氢呋喃(4毫升)中滴加氢氧化锂(43.0毫克，1.0毫摩尔)水溶液(2毫升)，滴毕，室温反应45分钟。Under an ice bath, add methyl 2-((5,6-dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate (150.0 mg, 0.5 mmol) Lithium hydroxide (43.0 mg, 1.0 mmol) aqueous solution (2 mL) was added dropwise to the tetrahydrofuran (4 mL). After the dripping was completed, the reaction was carried out at room temperature for 45 minutes.
反应结束，加水(5毫升)稀释，用稀盐酸水溶液(1.0摩尔/升)调pH至3，混合液用乙酸乙酯(20毫升×3次)萃取，合并有机相，然后用无水硫酸钠干燥，最后减压浓缩得到140.0毫克黄色固体2-((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸，无需纯化，直接用于下一步反应。LCMS:RT＝1.06min,[M-H] -＝299.98。 After the reaction is over, dilute with water (5 ml), adjust the pH to 3 with dilute aqueous hydrochloric acid (1.0 mol/L), extract the mixture with ethyl acetate (20 ml × 3 times), combine the organic phases, and then use anhydrous sodium sulfate Dry, and finally concentrate under reduced pressure to obtain 140.0 mg of yellow solid 2-((5,6-dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid without purification, directly Used in the next reaction. LCMS: RT=1.06min, [MH] - =299.98.
步骤E：合成(S)-4-(2-(2-(((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-(((5,6-Dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000135
Figure PCTCN2020133493-appb-000135
室温下，将2-((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(140.0毫克，0.5毫摩尔)、(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(174.0毫克，0.5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(263.0毫克，0.7毫摩尔)加入N,N-二甲基甲酰胺(2.0毫升)中，滴加N,N-二异丙基乙胺(120.0毫克，0.9毫摩尔)，滴毕，N 2保护下，室温反应3小时。 At room temperature, 2-((5,6-dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (140.0 mg, 0.5 mmol), (S) Tert-Butyl-4-(2-amino-3-phenylpropionamido)benzoate (174.0 mg, 0.5 mmol) and 2-(7-benzotriazole oxide)-N,N,N', N'-tetramethylurea hexafluorophosphate (263.0 mg, 0.7 mmol) was added to N,N-dimethylformamide (2.0 mL), and N,N-diisopropylethylamine (120.0 mg) was added dropwise , 0.9 mmol), after dripping, under N 2 protection, react at room temperature for 3 hours.
反应结束，加入乙酸乙酯(10毫升)，混合液用饱和食盐水(10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/正己烷＝1/5)。得到255.0毫克白色固体(S)-4-(2-(2-(((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：88.9％)。LCMS:RT＝4.19min,[M-H] -＝624.15。 After the reaction was completed, ethyl acetate (10 mL) was added, and the mixture was washed with 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/n-hexane=1/5). Obtained 255.0 mg of white solid (S)-4-(2-(2-(((5,6-dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-phenylpropionamido) tert-butyl benzoate (yield: 88.9%). LCMS: RT = 4.19 min, [MH] - =624.15.
步骤F：合成(S)-4-(2-(2-(((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-(((5,6-Dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000136
Figure PCTCN2020133493-appb-000136
室温下，将(S)-4-(2-(2-(((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(80.0毫克，0.3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0.5小时。At room temperature, the (S)-4-(2-(2-(((5,6-dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide (4)-3-phenylpropionamido) tert-butyl benzoate (80.0 mg, 0.3 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 0.5 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸，所得残余物用制备液相色谱仪分离，得到8.4毫克白色固体(S)-4-(2-(2-(((5,6-二氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：5.5％)。LCMS:RT＝3.58min,[M+H] +＝568.08。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump. The residue obtained is separated by a preparative liquid chromatograph to obtain 8.4 mg of white solid (S)-4-(2-(2-(((5,6 -Dichloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid (yield: 5.5%). LCMS :RT＝3.58min, [M+H] + ＝568.08.
实施例15Example 15
合成(S)-4-(2-(2-(((5-氯-2-(环丙基甲氧基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(cyclopropylmethoxy)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamide Base) benzoic acid
Figure PCTCN2020133493-appb-000137
Figure PCTCN2020133493-appb-000137
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-氯-2-(环丙基甲氧基)硝基苯Step A: Synthesis of 5-chloro-2-(cyclopropylmethoxy)nitrobenzene
Figure PCTCN2020133493-appb-000138
Figure PCTCN2020133493-appb-000138
室温下，将5-氯-2-氟硝基苯(1.8克，10.3毫摩尔)、环丙基甲醇(879.0毫克，12.2毫摩尔)和碳酸铯(6.8克，20.9毫摩尔)加入四氢呋喃(20.0毫升)中，N 2保护下，室温反应6小时。 At room temperature, add 5-chloro-2-fluoronitrobenzene (1.8 g, 10.3 mmol), cyclopropyl methanol (879.0 mg, 12.2 mmol) and cesium carbonate (6.8 g, 20.9 mmol) to tetrahydrofuran (20.0 ML), under the protection of N 2 , react at room temperature for 6 hours.
反应结束，加入饱和食盐水50毫升，用乙酸乙酯(50毫升×3次)萃取，然后用无水硫酸镁干燥，最后减压浓缩得到2.1克金色油状物4-氯-1-(环丙基甲氧基)-2-硝基苯，无需纯化，直接用于下一步反应。After the reaction was completed, 50 ml of saturated brine was added, extracted with ethyl acetate (50 ml × 3 times), then dried over anhydrous magnesium sulfate, and finally concentrated under reduced pressure to obtain 2.1 g of golden oily 4-chloro-1-(cyclopropyl) (Methoxy)-2-nitrobenzene, without purification, used directly in the next reaction.
步骤B：合成5-氯-2-(环丙基甲氧基)苯胺Step B: Synthesis of 5-chloro-2-(cyclopropylmethoxy)aniline
Figure PCTCN2020133493-appb-000139
Figure PCTCN2020133493-appb-000139
室温下，5-氯-2-(环丙基甲氧基)硝基苯(2.1克，9.3毫摩尔)加入乙酸乙酯(100毫升)中，冰浴下，分批加入氯化亚锡二水合物(23.6克，10.5毫摩尔)，N 2保护下，室温搅拌过夜。 At room temperature, 5-chloro-2-(cyclopropylmethoxy)nitrobenzene (2.1 g, 9.3 mmol) was added to ethyl acetate (100 ml), and stannous chloride was added in batches under ice bath. The hydrate (23.6 g, 10.5 mmol) was stirred overnight at room temperature under the protection of N 2.
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50毫升乙酸乙酯洗涤，滤液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(50毫升×2次)洗涤，然后用无水硫酸镁干燥，最后减压浓缩。得到1.2克黄色油状5-氯-2-(环丙基甲氧基)苯胺，粗品直接用于下一步反应。LCMS:RT＝3.99min,[M+H] +＝198.12。 After the reaction was completed, it was quenched with saturated sodium bicarbonate aqueous solution, and excess sodium bicarbonate was added to adjust the pH to weakly alkaline. The filter cake was filtered with Celite pad and the filter cake was washed with 50 ml of ethyl acetate. The filtrate was washed with ethyl acetate ( 30 ml × 3 times), and the organic phases were combined. The organic phase was washed with saturated brine (50 ml × 2 times), then dried over anhydrous magnesium sulfate, and finally concentrated under reduced pressure. 1.2 g of yellow oily 5-chloro-2-(cyclopropylmethoxy)aniline was obtained, and the crude product was directly used in the next reaction. LCMS: RT = 3.99 min, [M+H] + =198.12.
步骤C：合成2-((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000140
Figure PCTCN2020133493-appb-000140
N 2保护、冰浴下，向含有5-氯-2-(环丙基甲氧基)苯胺(1.2克，5.9毫摩尔)和吡啶(948.0毫克，12.0毫摩尔)的二氯甲烷(25毫升)中滴加草酰氯单甲酯(878.0毫克，7.2毫摩尔)，滴毕，室温反应1.5小时。 Under N 2 protection and ice bath, add 5-chloro-2-(cyclopropylmethoxy)aniline (1.2 g, 5.9 mmol) and pyridine (948.0 mg, 12.0 mmol) to dichloromethane (25 mL). ) Was added dropwise to monomethyl oxalyl chloride (878.0 mg, 7.2 mmol), after dropping, the reaction was carried out at room temperature for 1.5 hours.
反应结束，加饱和碳酸氢钠溶液淬灭，混合液用二氯甲烷(50毫升×3次)萃取。合并有机相，有机相用无水硫酸钠干燥，最后减压浓缩得到2.1克棕色固体2-((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酸甲酯粗品，无需纯化，直接用于下一步反应。LCMS:RT＝3.43min,[M-CH 3] -＝268.10。 After the reaction was over, it was quenched by adding saturated sodium bicarbonate solution, and the mixture was extracted with dichloromethane (50 ml×3 times). The organic phases were combined, dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure to obtain 2.1 g of brown solid 2-((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetic acid The crude methyl ester was used directly in the next reaction without purification. LCMS: RT = 3.43 min, [M-CH 3 ] - =268.10.
步骤D：合成2-((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酸Step D: Synthesis of 2-((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000141
Figure PCTCN2020133493-appb-000141
冰浴下，向含有2-((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酸甲酯(2.1克，7.4毫摩尔)的四氢呋喃(10毫升)中滴加氢氧化锂(0.7克，16.2毫摩尔)水溶液(5毫升)，滴毕后升至室温搅拌3.0小时。Under ice bath, add methyl 2-((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetate (2.1g, 7.4mmol) in tetrahydrofuran (10ml) An aqueous solution (5 ml) of lithium hydroxide (0.7 g, 16.2 mmol) was added dropwise to the mixture, and after the dropping was completed, it was raised to room temperature and stirred for 3.0 hours.
反应结束，加水稀释，用稀盐酸水溶液(1.0摩尔/升)调pH至3，混合液用乙酸乙酯(30毫升×3次)萃取，合并有机相，然后用无水硫酸钠干燥，最后减压浓缩得到623.0毫克棕黄色固体2-((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酸，无需纯化，直接用于下一步反应。LCMS:RT＝3.41min,[M-H] -＝268.10。 After the reaction is over, dilute with water, adjust the pH to 3 with dilute aqueous hydrochloric acid (1.0 mol/L), extract the mixture with ethyl acetate (30 ml×3 times), combine the organic phases, and then dry with anhydrous sodium sulfate, and finally reduce It was concentrated under pressure to obtain 623.0 mg of brown-yellow solid 2-((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetic acid, which was used directly in the next reaction without purification. LCMS: RT=3.41min, [MH] - =268.10.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000142
Figure PCTCN2020133493-appb-000142
室温下，将2-((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酸(117.0毫克，0.4毫摩尔)、(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(140.0毫克，0.4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(277.0毫克，0.7毫摩尔)加入N,N-二甲基甲酰胺(8.0毫升)中，滴加N,N-二异丙基乙胺(114.0毫克，0.8毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetic acid (117.0 mg, 0.4 mmol), (S)-4-(2- Amino-3-phenylpropionamido) tert-butyl benzoate (140.0 mg, 0.4 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyl Urea hexafluorophosphate (277.0 mg, 0.7 mmol) was added to N,N-dimethylformamide (8.0 mL), and N,N-diisopropylethylamine (114.0 mg, 0.8 mmol) was added dropwise, After dripping, under the protection of N 2 , react at room temperature overnight.
反应结束，加入25毫升乙酸乙酯，混合液用饱和食盐水(20毫升×3次)洗涤，然后有机相用无水硫酸钠干燥，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87.0毫克白色粉末固体(S)-4-(2-(2-(((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：33.9％)。LCMS:RT＝4.91min,[M+H] +＝611.18。 After the reaction was completed, 25 ml of ethyl acetate was added, the mixed solution was washed with saturated brine (20 ml×3 times), then the organic phase was 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=1/3). Obtain 87.0 mg of white powder solid (S)-4-(2-(2-(((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido) tert-butyl benzoate (yield: 33.9%). LCMS: RT = 4.91 min, [M+H] + =611.18.
步骤F：合成(S)-4-(2-(2-(((环丙基甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-(((cyclopropylmethyl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000143
Figure PCTCN2020133493-appb-000143
室温下，将(S)-4-(2-(2-(((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯叔丁酯(87.0毫克，0.1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0.5小时。At room temperature, the (S)-4-(2-(2-(((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxoacetamido)-3-benzene (87.0 mg, 0.1 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 0.5 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(3.0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65.3毫克白色固体(S)-4-(2-(2-(((5-氯-2-(环丙基甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：83.2％)。LCMS:RT＝4.18min,[M-H] -＝579.53。 After the reaction was completed, the dichloromethane was evaporated and the trifluoroacetic acid was drained by an oil pump. The obtained residue was dissolved in dichloromethane (3.0 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 65.3 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(cyclopropylmethyl)phenyl)amino)-2-oxo (Alsoacetamido)-3-phenylpropionamido)benzoic acid (yield: 83.2%). LCMS: RT = 4.18 min, [MH] - = 579.53.
实施例16Example 16
合成(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((3-氧代异吲哚-5-基)氨基)-3-苯基丙烷-2-基)草酰胺 Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((3-oxoisoindole- 5-yl)amino)-3-phenylpropan-2-yl)oxamide
Figure PCTCN2020133493-appb-000144
Figure PCTCN2020133493-appb-000144
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-2-氨基-N-(1-氧代异吲哚-5-基)-3-苯基丙酰胺Step A: Synthesis of (S)-2-amino-N-(1-oxoisoindol-5-yl)-3-phenylpropionamide
Figure PCTCN2020133493-appb-000145
Figure PCTCN2020133493-appb-000145
室温下，将(叔丁氧羰基)-L-苯丙氨酸(200毫克，0.75毫摩尔)、6-氨基异吲哚-1-酮(112毫克，0.75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(430克，1.13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(195毫克，1.5毫摩尔)，滴毕，N 2保护下，室温反应过夜。反应结束，倒入40毫升水中，固体析出，抽滤，干燥得白色固体直接用于下一步反应。 At room temperature, (tert-butoxycarbonyl)-L-phenylalanine (200 mg, 0.75 mmol), 6-aminoisoindol-1-one (112 mg, 0.75 mmol) and 2-(7- Oxidized benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (430g, 1.13mmol) was added to N,N-dimethylformamide (4.0ml), N,N-diisopropylethylamine (195 mg, 1.5 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. After the reaction was completed, it was poured into 40 ml of water. The solid was separated out, filtered and dried to obtain a white solid which was directly used in the next reaction.
将上述粗产物溶于5毫升HCl/EA溶液(2摩尔/升)，室温搅拌3小时，反应完全。抽滤，并用少量EA洗一次，干燥得121毫克白色固体(S)-2-氨基-N-(3-氧代异吲哚-5-基)-3-苯基丙酰胺。MS(ESI)M/Z：296.18[M+H] +。 The above crude product was dissolved in 5 ml of HCl/EA solution (2 mol/L) and stirred at room temperature for 3 hours, and the reaction was complete. It was filtered with suction, washed once with a small amount of EA, and dried to obtain 121 mg of white solid (S)-2-amino-N-(3-oxoisoindol-5-yl)-3-phenylpropionamide. MS (ESI) M/Z: 296.18 [M+H] + .
步骤B：合成(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((3-氧代异吲哚-5-基)氨基)-3-苯基丙烷-2-基)草酰胺 Step B: Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((3-oxoiso Indol-5-yl)amino)-3-phenylpropan-2-yl)oxalamide
Figure PCTCN2020133493-appb-000146
Figure PCTCN2020133493-appb-000146
室温下，将2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸(80毫克，0.30毫摩尔)、(S)-2-氨基-N-(1-氧代异吲哚-5-基)-3-苯基丙酰胺(100毫克，0.34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(171克，0.45毫摩尔)加入N,N-二甲基甲酰胺(2.0毫升)中，滴加N,N-二异丙基乙胺(116毫克，0.90毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetic acid (80 mg, 0.30 mmol), (S)-2-Amino-N-(1-oxoisoindol-5-yl)-3-phenylpropionamide (100 mg, 0.34 mmol) and 2-(7-oxobenzotriazole) )-N,N,N',N'-Tetramethylurea hexafluorophosphate (171g, 0.45mmol) was added to N,N-dimethylformamide (2.0ml), and N,N- Diisopropylethylamine (116 mg, 0.90 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。所得残余物用制备HPLC纯化。得到73毫克白色固体(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((3-氧代异吲哚-5-基)氨基)-3-苯基丙烷-2-基)草酰胺(收率：44.6％)。MS(ESI)M/Z：545.19[M+H] +。 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 resulting residue was purified by preparative HPLC. Obtained 73 mg of white solid (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((3-oxo Isoindol-5-yl)amino)-3-phenylpropan-2-yl)oxamide (yield: 44.6%). MS (ESI) M/Z: 545.19 [M+H] + .
实施例17Example 17
合成(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((1-氧代异吲哚-5-基)氨基)-3-苯基丙烷-2-基)草酰胺 Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((1-oxoisoindole- 5-yl)amino)-3-phenylpropan-2-yl)oxamide
Figure PCTCN2020133493-appb-000147
Figure PCTCN2020133493-appb-000147
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-2-氨基-N-(1-氧代异吲哚-5-基)-3-苯基丙酰胺Step A: Synthesis of (S)-2-amino-N-(1-oxoisoindol-5-yl)-3-phenylpropionamide
Figure PCTCN2020133493-appb-000148
Figure PCTCN2020133493-appb-000148
室温下，将(叔丁氧羰基)-L-苯丙氨酸(200毫克，0.75毫摩尔)、5-氨基异吲哚-1-酮(112毫克，0.75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(430克，1.13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(195毫克，1.5毫摩尔)，滴毕，N 2保护下，室温反应过夜。反应结束，倒入40毫升水中，固体析出，抽滤，干燥得白色固体直接用于下一步反应。 At room temperature, (tert-butoxycarbonyl)-L-phenylalanine (200 mg, 0.75 mmol), 5-aminoisoindol-1-one (112 mg, 0.75 mmol) and 2-(7- Oxidized benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (430g, 1.13mmol) was added to N,N-dimethylformamide (4.0ml), N,N-diisopropylethylamine (195 mg, 1.5 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. After the reaction was completed, it was poured into 40 ml of water. The solid was separated out, filtered and dried to obtain a white solid which was directly used in the next reaction.
将上述粗产物溶于5毫升HCl/EA溶液(2摩尔/升)，室温搅拌3h，反应完全。抽滤，并用少量EA洗一次，干燥得110毫克白色固体(S)-2-氨基-N-(1-氧代异吲哚-5-基)-3-苯基丙酰胺。MS(ESI)M/Z：296.16[M+H] +。 The above crude product was dissolved in 5 ml of HCl/EA solution (2 mol/L), stirred at room temperature for 3 hours, and the reaction was complete. It was filtered with suction, washed once with a small amount of EA, and dried to obtain 110 mg of white solid (S)-2-amino-N-(1-oxoisoindol-5-yl)-3-phenylpropionamide. MS (ESI) M/Z: 296.16 [M+H] + .
步骤B：合成(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((1-氧代异吲哚-5-基)氨基)-3-苯基丙烷-2-基)草酰胺 Step B: Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((1-oxoiso Indol-5-yl)amino)-3-phenylpropan-2-yl)oxalamide
Figure PCTCN2020133493-appb-000149
Figure PCTCN2020133493-appb-000149
室温下，将2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸(80毫克，0.30毫摩尔)、(S)-2-氨基-N-(1-氧代异吲哚-5-基)-3-苯基丙酰胺(100毫克，0.34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(171克，0.45毫摩尔)加入N,N-二甲基甲酰胺(2.0毫升)中，滴加N,N-二异丙基乙胺(116毫克，0.90毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetic acid (80 mg, 0.30 mmol), (S)-2-Amino-N-(1-oxoisoindol-5-yl)-3-phenylpropionamide (100 mg, 0.34 mmol) and 2-(7-oxobenzotriazole) )-N,N,N',N'-Tetramethylurea hexafluorophosphate (171g, 0.45mmol) was added to N,N-dimethylformamide (2.0ml), and N,N- Diisopropylethylamine (116 mg, 0.90 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。所得残余物用制备HPLC纯化。得到70毫克白色固体(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((1-氧代异吲哚-5-基)氨基)-3-苯基丙烷-2-基)草酰胺(收率：42.8％)。MS(ESI)M/Z：545.18[M+H] +。 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 resulting residue was purified by preparative HPLC. Obtain 70 mg of white solid (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((1-oxo Isoindol-5-yl)amino)-3-phenylpropan-2-yl)oxamide (yield: 42.8%). MS (ESI) M/Z: 545.18 [M+H] + .
实施例18Example 18
合成(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((2-氧代-2,3-二氢-1H-苯并[d]咪唑-5-基)氨基)-3-苯基丙-2-基)草酰胺 Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((2-oxo-2,3 -Dihydro-1H-benzo[d]imidazol-5-yl)amino)-3-phenylprop-2-yl)oxamide
Figure PCTCN2020133493-appb-000150
Figure PCTCN2020133493-appb-000150
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-2-氨基-N-(2-氧代-2,3-二氢-1H-苯并[d]咪唑-5-基)-3-苯基丙酰胺Step A: Synthesis of (S)-2-amino-N-(2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-3-phenylpropionamide
Figure PCTCN2020133493-appb-000151
Figure PCTCN2020133493-appb-000151
室温下，将(叔丁氧羰基)-L-苯丙氨酸(200毫克，0.75毫摩尔)、6-氨基异吲哚-1-酮(113毫克，0.75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(430克，1.13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(195毫克，1.5毫摩尔)，滴毕，N 2保护下，室温反应过夜。反应结束，倒入40毫升水中，固体析出，抽滤，干燥得白色固体直接用于下一步反应。 At room temperature, (tert-butoxycarbonyl)-L-phenylalanine (200 mg, 0.75 mmol), 6-aminoisoindol-1-one (113 mg, 0.75 mmol) and 2-(7- Oxidized benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (430g, 1.13mmol) was added to N,N-dimethylformamide (4.0ml), N,N-diisopropylethylamine (195 mg, 1.5 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. After the reaction was completed, it was poured into 40 ml of water. The solid was separated out, filtered and dried to obtain a white solid which was directly used in the next reaction.
将上述粗产物溶于5毫升HCl/EA溶液(2摩尔/升)，室温搅拌3小时，反应完全。抽滤，并用少量EA洗一次，干燥得125毫克白色固体(S)-2-氨基-N-(2-氧代-2,3-二氢-1H-苯并[d]咪唑-5-基)-3-苯基丙酰胺。MS(ESI)M/Z：297.15[M+H] +。 The above crude product was dissolved in 5 ml of HCl/EA solution (2 mol/L) and stirred at room temperature for 3 hours, and the reaction was complete. Filter with suction, wash once with a small amount of EA, and dry to obtain 125 mg of white solid (S)-2-amino-N-(2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl )-3-Phenylpropionamide. MS (ESI) M/Z: 297.15 [M+H] + .
步骤B：合成(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((2-氧代-2,3-二氢-1H-苯并[d]咪唑-5-基)氨基)-3-苯基丙-2-基)草酰胺 Step B: Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((2-oxo- 2,3-Dihydro-1H-benzo[d]imidazol-5-yl)amino)-3-phenylprop-2-yl)oxamide
Figure PCTCN2020133493-appb-000152
Figure PCTCN2020133493-appb-000152
室温下，将2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸(80毫克，0.30毫摩尔)、(S)-2-氨基-N-(2-氧代-2,3-二氢-1H-苯并[d]咪唑-5-基)-3-苯基丙酰胺(110毫克，0.37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(171克，0.45毫摩尔)加入N,N-二甲基甲酰胺(2.0毫升)中，滴加N,N-二异丙基乙胺(116毫克，0.90毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetic acid (80 mg, 0.30 mmol), (S)-2-Amino-N-(2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)-3-phenylpropionamide (110 mg, 0.37 mmol ) And 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (171g, 0.45mmol) add N,N-dimethylformaldehyde To the amide (2.0 ml), N,N-diisopropylethylamine (116 mg, 0.90 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(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。所得残余物用制备HPLC纯化。得到68毫克白色固体(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((2-氧代-2,3-二氢-1H-苯并[d]咪唑-5-基)氨基)-3-苯基丙-2-基)草酰胺(收率：41.6％)。MS(ESI)M/Z：546.14[M+H] +。 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 resulting residue was purified by preparative HPLC. Obtain 68 mg of white solid (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((2-oxo -2,3-Dihydro-1H-benzo[d]imidazol-5-yl)amino)-3-phenylprop-2-yl)oxamide (yield: 41.6%). MS (ESI) M/Z: 546.14 [M+H] + .
实施例19Example 19
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000153
Figure PCTCN2020133493-appb-000153
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸甲酯Step A: Synthesis of methyl (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido)benzoate
Figure PCTCN2020133493-appb-000154
Figure PCTCN2020133493-appb-000154
室温下，将(S)-3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酸(2.5克，7.3毫摩尔)、4-氨基苯甲酸甲酯(1.1克，7.3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(4.15克，10.9毫摩尔)加入N,N-二甲基甲酰胺(25毫升)中，滴加N,N-二异丙基乙胺(2.8克，21.7毫摩尔)，滴毕，N 2保护下，室温反应16h。 At room temperature, (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid (2.5 g, 7.3 mmol), methyl 4-aminobenzoate (1.1 g , 7.3 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (4.15 g, 10.9 mmol) were added to N,N- In dimethylformamide (25 mL), N,N-diisopropylethylamine (2.8 g, 21.7 mmol) was added dropwise, after the dripping, the reaction was carried out at room temperature under N 2 protection for 16 h.
反应结束，倒入30毫升水中，固体析出，抽滤，干燥得3.5克白色固体(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸甲酯(收率100％)。MS(ESI)M/Z:476.20[M+H] +。 After the reaction is over, pour it into 30 ml of water. The solid is separated out, filtered with suction and dried to obtain 3.5 g of white solid (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propyl Amido) methyl benzoate (yield 100%). MS (ESI) M/Z: 476.20 [M+H] + .
步骤B：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸甲酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido ) Methyl benzoate
Figure PCTCN2020133493-appb-000155
Figure PCTCN2020133493-appb-000155
氮气保护下，向含有(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸甲酯(2.0克，4.19毫摩尔)、4-甲基哌嗪-2-酮(956毫克，8.38毫摩尔)的二氧六环(25毫升)溶液中加入碘化亚铜(797毫克，4.19毫摩尔)，碳酸铯(2.73克，8.38毫摩尔)，N 1,N 2-二甲基乙二胺(738毫克，8.38毫摩尔)，加毕，加热至120摄氏度回流反应过夜。 Under the protection of nitrogen, add methyl (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido)benzoate (2.0 g, 4.19 mmol) , 4-methylpiperazine-2-one (956 mg, 8.38 mmol) in dioxane (25 mL) was added cuprous iodide (797 mg, 4.19 mmol), cesium carbonate (2.73 g, 8.38 mmol), N 1 ,N 2 -dimethylethylenediamine (738 mg, 8.38 mmol), after the addition, heat to 120 degrees Celsius and reflux for overnight reaction.
反应结束，加水(20毫升)淬灭，乙酸乙酯萃取(150毫升×2次)，合并有机相，用饱和食盐水(100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：正己烷/乙酸乙酯＝9/1)。得到900毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸甲酯(42.1％)。MS(ESI)M/Z:511.26[M+H] +。 After the reaction was completed, it was quenched by adding water (20 mL), extracted with ethyl acetate (150 mL×2 times), and the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane/ethyl acetate=9/1). Obtain 900 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamide Methyl)benzoate (42.1%). MS (ESI) M/Z: 511.26 [M+H] + .
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸甲酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)methyl benzoate
Figure PCTCN2020133493-appb-000156
Figure PCTCN2020133493-appb-000156
室温下，将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸甲酯(900毫克，1.76毫摩尔)溶于盐酸/乙酸乙酯溶液(12毫升，2摩尔/升)，室温反应3小时。抽滤，干燥，得1.0克淡黄色固体(S)-4-(2-氨基-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸甲酯二盐酸盐。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido ) Methyl benzoate (900 mg, 1.76 mmol) was dissolved in a hydrochloric acid/ethyl acetate solution (12 mL, 2 mol/L) and reacted at room temperature for 3 hours. Filtration with suction and drying to obtain 1.0 g of light yellow solid (S)-4-(2-amino-3-(4-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido) Methyl benzoate dihydrochloride.
室温下，将2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸(268毫克，1.0毫摩尔)、(S)-4-(2-氨基-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸甲酯二盐酸盐(507毫克，1.05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(570毫克，1.5毫摩尔)加入N,N-二甲基甲酰胺(8.0毫升)中，滴加N,N-二异丙基乙胺(516毫克，4.0毫摩尔)，滴毕，N 2保护下，室温反应过夜。反应结束，倒入20毫升水中，固体析出，抽滤，干燥，得到360毫克淡黄色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸甲酯(收率：54.5％)。MS(ESI)M/Z:660.30[M+H] +。 At room temperature, 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetic acid (268 mg, 1.0 mmol), (S)-4-(2-Amino-3-(4-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid methyl ester dihydrochloride (507 mg , 1.05 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (570 mg, 1.5 mmol) was added to N,N- In dimethylformamide (8.0 ml), N,N-diisopropylethylamine (516 mg, 4.0 mmol) was added dropwise, and after dropping, the reaction was carried out at room temperature overnight under the protection of N 2. After the reaction is over, it is poured into 20 ml of water, the solid is separated out, filtered with suction, and dried to obtain 360 mg of light yellow solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazole-1) -Yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido)methyl benzoate (Yield: 54.5%). MS (ESI) M/Z: 660.30 [M+H] + .
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000157
Figure PCTCN2020133493-appb-000157
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸甲酯(330毫克，0.50毫摩尔)溶于四氢呋喃/水溶液(10毫升/5毫升)，加入一水合氢氧化锂(42毫克，1.0毫摩尔)，加热至40摄氏度反应过夜。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)methyl benzoate (330mg, 0.50mmol) dissolved in tetrahydrofuran/water solution (10ml/5ml ), add lithium hydroxide monohydrate (42 mg, 1.0 mmol), and heat to 40 degrees Celsius to react overnight.
反应完全后减压蒸除有机溶剂，用2摩尔/升盐酸调节pH至7-8，析出固体，抽滤，烘干得240毫克淡黄色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸(收率：74.4％)。MS(ESI)M/Z:644.10[M-H] -。 After the completion of the reaction, the organic solvent was evaporated under reduced pressure, and the pH was adjusted to 7-8 with 2 mol/L hydrochloric acid. A solid was precipitated, filtered with suction, and dried to obtain 240 mg of light yellow solid (S)-4-(2-(2-() (5-Chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-methyl-2-oxopiperazine-1 -Yl)phenyl)propionamido)benzoic acid (yield: 74.4%). MS(ESI) M/Z: 644.10 [MH] - .
实施例20Example 20
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰胺基)-3-(吡啶-3-基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(pyridine- 3-yl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000158
Figure PCTCN2020133493-appb-000158
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(吡啶-3-基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of tert-butyl (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(pyridin-3-yl)propionamido)benzoate
Figure PCTCN2020133493-appb-000159
Figure PCTCN2020133493-appb-000159
室温下，将(S)-2-((叔丁氧羰基)氨基)-3-(吡啶-3-基)丙酸(532毫克，2.0毫摩尔)、4-氨基苯甲酸叔丁酯(386毫克，2.0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(1.14克，3.0毫摩尔)加入N,N-二甲基乙酰胺(8毫升)中，滴加N,N-二异丙基乙胺(774毫克，6.0毫摩尔)，滴毕，N 2保护下，室温反应16h。 At room temperature, (S)-2-((tert-butoxycarbonyl)amino)-3-(pyridin-3-yl)propionic acid (532 mg, 2.0 mmol), tert-butyl 4-aminobenzoate (386 Mg, 2.0 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.14 g, 3.0 mmol) add N,N -In dimethylacetamide (8 mL), add N,N-diisopropylethylamine (774 mg, 6.0 mmol) dropwise, after dropping, under N 2 protection, react at room temperature for 16 h.
反应结束，倒入20毫升水中，固体析出，抽滤，干燥得882克白色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(吡啶-3-基)丙酰胺基)苯甲酸叔丁酯(收率100％)。MS(ESI)M/Z:442.25[M+H] +。 After the reaction is over, it is poured into 20 ml of water. The solid is separated out, filtered and dried to obtain 882 g of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(pyridin-3-yl) Propionamido) tert-butyl benzoate (yield 100%). MS (ESI) M/Z: 442.25 [M+H] + .
步骤B：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(吡啶-3-基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(Pyridin-3-yl) propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000160
Figure PCTCN2020133493-appb-000160
室温下，将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(吡啶-3-基)丙酰胺基)苯甲酸叔丁酯(882毫克，2.0毫摩尔)溶于乙酸乙酯(15毫升)，向该溶液加入盐酸/乙酸乙酯溶液(8毫升，2摩尔/升)，室温反应2小时。抽滤，干燥，得800毫克淡黄色固体(S)-4-(2-氨基-3-(吡啶-3-基)丙酰胺基)苯甲酸叔丁酯。At room temperature, (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(pyridin-3-yl)propionamido) tert-butyl benzoate (882 mg, 2.0 mmol) Dissolve in ethyl acetate (15 mL), add hydrochloric acid/ethyl acetate solution (8 mL, 2 mol/L) to the solution, and react at room temperature for 2 hours. It was filtered with suction and dried to obtain 800 mg of tert-butyl (S)-4-(2-amino-3-(pyridin-3-yl)propionamido)benzoate as a pale yellow solid.
室温下，将2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸(120毫克，0.45毫摩尔)、(S)-4-(2-氨基-3-(4-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(205毫克，0.50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(257毫克，0.68毫摩尔)加入N,N-二甲基乙酰胺(3.0毫升)中，滴加N,N-二异丙基乙胺(232毫克，1.8毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetic acid (120 mg, 0.45 mmol), (S)-4-(2-Amino-3-(4-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (205 mg, 0.50 mg Mol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (257 mg, 0.68 mmol) was added to N,N-dimethyl In acetamide (3.0 ml), N,N-diisopropylethylamine (232 mg, 1.8 mmol) was added dropwise, and after dropping, the reaction was carried out at room temperature overnight under the protection of N 2.
反应结束，倒入20毫升水中，固体析出，抽滤，干燥，得到150毫克淡黄色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(吡啶-3-基)丙酰胺基)苯甲酸叔丁酯(收率：56.4％)。MS(ESI)M/Z:591.23[M+H] +。 After the reaction is over, it is poured into 20 ml of water, the solid is separated out, filtered with suction, and dried to obtain 150 mg of light yellow solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazole-1) -Yl)phenyl)amino)-2-oxoacetamido)-3-(pyridin-3-yl)propionamido)tert-butyl benzoate (yield: 56.4%). MS (ESI) M/Z: 591.23 [M+H] + .
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰胺基)-3-(吡啶-3-基)丙酰胺基)苯甲酸Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3- (Pyridin-3-yl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000161
Figure PCTCN2020133493-appb-000161
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(吡啶-3-基)丙酰胺基)苯甲酸叔丁酯(80毫克，0.14毫摩尔)溶于二氯甲烷(2.0毫升)中，加入三乙基硅烷(40毫克，0.34毫摩尔)，缓慢滴加三氟乙酸(0.4毫升)，室温反应过夜。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(Pyridin-3-yl) propionamido) tert-butyl benzoate (80 mg, 0.14 mmol) was dissolved in dichloromethane (2.0 mL), and triethylsilane (40 mg, 0.34 mmol) was added, Slowly add trifluoroacetic acid (0.4 ml) dropwise, and react at room temperature overnight.
反应结束，减压蒸干溶剂，粗产物进行制备HPLC，得53毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰胺基)-3-(吡啶-3-基)丙酰胺基)苯甲酸(收率73.4％)。MS(ESI)M/Z:533.09[M-H] -。 After the reaction, the solvent was evaporated under reduced pressure, and the crude product was subjected to preparative HPLC to obtain 53 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)) Phenyl)amino)-2-oxoacetamido)-3-(pyridin-3-yl)propionamido)benzoic acid (yield 73.4%). MS(ESI) M/Z: 533.09 [MH] - .
实施例21Example 21
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenyl Propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000162
Figure PCTCN2020133493-appb-000162
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-((叔丁氧基羰基)氨基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸乙酯Step A: Synthesis of (S)-5-(2-((tert-butoxycarbonyl)amino)-3-phenylpropionamido)-1H-indole-2-carboxylic acid ethyl ester
Figure PCTCN2020133493-appb-000163
Figure PCTCN2020133493-appb-000163
室温下，将(叔丁氧羰基)-L-苯丙氨酸(265毫克，1.0毫摩尔)、5-氨基-1H-吲哚-2-羧酸乙酯(204毫克，1.0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(570毫克，1.5毫摩尔)加入N,N-二甲基甲酰胺(5毫升)中，滴加N,N-二异丙基乙胺(387毫克，3.0毫摩尔)，滴毕，N 2保护下，室温反应16h。 At room temperature, (tert-butoxycarbonyl)-L-phenylalanine (265 mg, 1.0 mmol), 5-amino-1H-indole-2-carboxylic acid ethyl ester (204 mg, 1.0 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (570 mg, 1.5 mmol) was added to N,N-dimethylformamide ( 5 ml), N,N-diisopropylethylamine (387 mg, 3.0 mmol) was added dropwise, after the dropping, the reaction was carried out at room temperature for 16 h under the protection of N 2.
反应结束，倒入80毫升饱和食盐水中，固体析出，抽滤，干燥得450毫克灰白色固体(S)-5-(2-((叔丁氧基羰基)氨基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸乙酯(收率100％)。MS(ESI)M/Z:452.25[M+H] +。 After the reaction is over, it is poured into 80 ml of saturated brine, the solid is separated out, filtered with suction, and dried to obtain 450 mg of off-white solid (S)-5-(2-((tert-butoxycarbonyl)amino)-3-phenylpropionamido ) Ethyl -1H-indole-2-carboxylate (yield 100%). MS (ESI) M/Z: 452.25 [M+H] + .
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸乙酯Step B: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid ethyl ester
Figure PCTCN2020133493-appb-000164
Figure PCTCN2020133493-appb-000164
室温下，将(S)-5-(2-(((叔丁氧基羰基)氨基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸乙酯(450毫克，1.0毫摩尔)溶于乙酸乙酯(5毫升)，向其中加入盐酸/乙酸乙酯溶液(5毫升，2摩尔/升)，室温反应4小时。抽滤，干燥，得110毫克淡黄色固体(S)-5-(2-氨基-3-苯基丙酰胺基)-1H-吲哚-2-羧酸乙酯盐酸盐。At room temperature, (S)-5-(2-(((tert-butoxycarbonyl)amino)-3-phenylpropionamido)-1H-indole-2-carboxylic acid ethyl ester (450 mg, 1.0 Millimoles) was dissolved in ethyl acetate (5 ml), hydrochloric acid/ethyl acetate solution (5 ml, 2 mol/L) was added to it, and reacted at room temperature for 4 hours. Filtered with suction and dried to obtain 110 mg of light yellow solid (S )-5-(2-Amino-3-phenylpropionamido)-1H-indole-2-carboxylic acid ethyl ester hydrochloride.
室温下，将2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸(70毫克，0.26毫摩尔)、(S)-5-(2-氨基-3-苯基丙酰胺基)-1H-吲哚-2-羧酸乙酯盐酸盐(110毫克，0.26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(148毫克，0.39毫摩尔)加入N,N-二甲基甲酰胺(2.0毫升)中，滴加N,N-二异丙基乙胺(134毫克，1.04毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetic acid (70 mg, 0.26 mmol), (S)-5-(2-Amino-3-phenylpropionamido)-1H-indole-2-carboxylic acid ethyl ester hydrochloride (110 mg, 0.26 mmol) and 2-(7-oxybenzene Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (148mg, 0.39mmol) was added to N,N-dimethylformamide (2.0ml) and added dropwise N,N-Diisopropylethylamine (134 mg, 1.04 mmol) was dripped and reacted overnight at room temperature under N 2 protection.
反应结束，倒入20毫升水中，固体析出，抽滤，干燥，得到150毫克淡黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸乙酯(收率：96.0％)。MS(ESI)M/Z:601.20[M+H] +。 After the reaction was completed, poured into 20 ml of water, solids separated out, filtered with suction, and dried to obtain 150 mg of pale yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazole-1) -Yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)-1H-indole-2-carboxylic acid ethyl ester (yield: 96.0%). MS (ESI) M/Z: 601.20 [M+H] + .
步骤C：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Step C: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000165
Figure PCTCN2020133493-appb-000165
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸乙酯(70毫克，0.12毫摩尔)溶于四氢呋喃/水溶液(3毫升/1.5毫升)，加入一水合氢氧化锂(10毫克，0.23毫摩尔)，加热至40摄氏度反应过夜。At room temperature, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid ethyl ester (70 mg, 0.12 mmol) was dissolved in tetrahydrofuran/water solution (3 ml/1.5 ml), and lithium hydroxide monohydrate (10 mg, 0.23 mmol), heated to 40 degrees Celsius and reacted overnight.
反应完全后减压蒸除有机溶剂，用2摩尔/升盐酸调节pH至5-6，析出固体，抽滤，粗品进行HPLC进一步纯化，得7毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：10.2％)。MS(ESI)M/Z:571.09[M-H] -。 After the reaction was completed, the organic solvent was distilled off under reduced pressure, and the pH was adjusted to 5-6 with 2 mol/L hydrochloric acid. The solid was precipitated and filtered with suction. The crude product was further purified by HPLC to obtain 7 mg of white solid (S)-5-(2-( 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)-1H-indole-2 -Carboxylic acid (yield: 10.2%). MS(ESI) M/Z: 571.09 [MH] - .
实施例22Example 22
合成(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((4-(5-氧代-2,5-二氢-1H-吡唑-3-基)苯基)氨基)-3-苯基丙-2-基)草酰胺 Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((4-(5-oxo- 2,5-Dihydro-1H-pyrazol-3-yl)phenyl)amino)-3-phenylprop-2-yl)oxamide
Figure PCTCN2020133493-appb-000166
Figure PCTCN2020133493-appb-000166
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-(4-硝基苯基)-1,2-二氢-3H-吡唑-3-酮Step A: Synthesis of 5-(4-nitrophenyl)-1,2-dihydro-3H-pyrazol-3-one
Figure PCTCN2020133493-appb-000167
Figure PCTCN2020133493-appb-000167
室温下，将3-(4-硝基苯基)-3氧代丙酸乙酯(237毫克，1.0毫摩尔)溶于乙醇(4毫升)，滴加一水合肼(94毫克，1.5毫摩尔)，加热至90摄氏度反应3小时。At room temperature, ethyl 3-(4-nitrophenyl)-3oxopropionate (237 mg, 1.0 mmol) was dissolved in ethanol (4 mL), and hydrazine monohydrate (94 mg, 1.5 mmol) was added dropwise. ), heated to 90 degrees Celsius and reacted for 3 hours.
反应完全后减压蒸除有机溶剂，所得固体加水(5毫升)打浆，抽滤，烘干，得183毫克淡红色固体5-(4-硝基苯基)-1,2-二氢-3H-吡唑-3-酮(收率：89.3％)。MS(ESI)M/Z:206.11[M+H] +。 After the reaction was completed, the organic solvent was evaporated under reduced pressure, the obtained solid was slurried with water (5 ml), filtered with suction, and dried to obtain 183 mg of pale red solid 5-(4-nitrophenyl)-1,2-dihydro-3H -Pyrazol-3-one (yield: 89.3%). MS(ESI) M/Z: 206.11 [M+H] + .
步骤B：合成5-(4-硝基苯基)-3-氧代-2,3-二氢-1H-吡唑-1-甲酸叔丁酯Step B: Synthesis of tert-butyl 5-(4-nitrophenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylate
Figure PCTCN2020133493-appb-000168
Figure PCTCN2020133493-appb-000168
室温下，向5-(4-硝基苯基)-1,2-二氢-3H-吡唑-3-酮(193毫克，0.75毫摩尔)的四氢呋喃/水(6毫升/3毫升)溶液中加入二碳酸二叔丁酯(409毫克，1.88毫摩尔)和氢氧化钠(75毫克，1.88毫摩尔)，室温反应过夜。To a solution of 5-(4-nitrophenyl)-1,2-dihydro-3H-pyrazol-3-one (193 mg, 0.75 mmol) in tetrahydrofuran/water (6 ml/3 ml) at room temperature Add di-tert-butyl dicarbonate (409 mg, 1.88 mmol) and sodium hydroxide (75 mg, 1.88 mmol), and react at room temperature overnight.
减压蒸除有机溶剂，残余物用乙酸乙酯(30毫升×2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(正己烷/乙酸乙酯＝4：1)，得到190毫克油状物5-(4-硝基苯基)-3-氧代-2,3-二氢-1H-吡唑-1-甲酸叔丁酯(收率：83.1％)。MS(ESI)M/Z:306.14[M+H] +。 The organic solvent was evaporated under reduced pressure, the residue was extracted with ethyl acetate (30 ml×2 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 (n-hexane/ethyl acetate=4:1) to obtain 190 mg of oily 5-(4-nitrophenyl)-3-oxo-2,3-dihydro- 1H-pyrazole-1-carboxylic acid tert-butyl ester (yield: 83.1%). MS (ESI) M/Z: 306.14 [M+H] + .
步骤C：合成5-(4-氨基苯基)-3-氧代-2,3-二氢-1H-吡唑-1-甲酸叔丁酯Step C: Synthesis of tert-butyl 5-(4-aminophenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylate
Figure PCTCN2020133493-appb-000169
Figure PCTCN2020133493-appb-000169
室温下，向5-(4-硝基苯基)-3-氧代-2,3-二氢-1H-吡唑-1-甲酸叔丁酯(150毫克，0.49毫摩尔)的甲醇(10毫升)溶液中加入钯/碳(30毫克)，体系置换为氢气，室温反应过夜。At room temperature, add 5-(4-nitrophenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylic acid tert-butyl ester (150 mg, 0.49 mmol) in methanol (10 (Ml) was added palladium/carbon (30 mg) to the solution, the system was replaced with hydrogen, and the reaction was carried out at room temperature overnight.
反应完全后，反应液经硅藻土过滤，滤饼用少量甲醇洗涤三次，合并滤液，减压蒸除有机溶剂，得到110毫克油状物5-(4-氨基苯基)-3-氧代-2,3-二氢-1H-吡唑-1-甲酸叔丁酯(收率：81.6％)。MS(ESI)M/Z:276.14[M+H] +。 After the reaction was completed, the reaction solution was filtered through Celite, the filter cake was washed three times with a small amount of methanol, the filtrates were combined, and the organic solvent was evaporated under reduced pressure to obtain 110 mg of oily 5-(4-aminophenyl)-3-oxo- Tert-butyl 2,3-dihydro-1H-pyrazole-1-carboxylate (yield: 81.6%). MS (ESI) M/Z: 276.14 [M+H] + .
步骤D：合成(S)-3-(4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯基)-5-氧代-2,5-二氢-1H-吡唑-1-羧酸叔丁酯Step D: Synthesis of (S)-3-(4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide (Phenyl)-3-phenylpropionamido)phenyl)-5-oxo-2,5-dihydro-1H-pyrazole-1-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000170
Figure PCTCN2020133493-appb-000170
室温下，向(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰基)-L-苯丙氨酸(42毫克，0.10毫摩尔)的二氯甲烷(2毫升)溶液中加入草酰氯(38毫克，0.30毫摩尔)，室温反应2小时。减压蒸干溶剂即得酰氯。At room temperature, to (2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetyl)-L-phenylalanine (42 mg, 0.10 mg Oxalyl chloride (38 mg, 0.30 mmol) was added to a dichloromethane (2 ml) solution of mol), and the reaction was carried out at room temperature for 2 hours. Evaporate the solvent under reduced pressure to get the acid chloride.
5-(4-氨基苯基)-3-氧代-2,3-二氢-1H-吡唑-1-甲酸叔丁酯(34毫克，0.12毫摩尔)溶于二氯甲烷(3毫升)，加入吡啶(24毫克，0.30毫摩尔)，将该溶液以冰水浴冷却，滴加上述酰氯的二氯甲烷(2毫升)溶液，加毕移除冷浴，室温反应2小时。5-(4-Aminophenyl)-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylic acid tert-butyl ester (34 mg, 0.12 mmol) dissolved in dichloromethane (3 mL) , Pyridine (24 mg, 0.30 mmol) was added, the solution was cooled in an ice-water bath, and the dichloromethane (2 ml) solution of the above acid chloride was added dropwise, the cooling bath was removed after the addition, and the reaction was carried out at room temperature for 2 hours.
加水(5毫升)淬灭，用乙酸乙酯(20毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)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(正己烷/乙酸乙酯＝1：2)，得到24毫克油状物(S)-3-(4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯基)-5-氧代-2,5-二氢-1H-吡唑-1-羧酸叔丁酯(收率：35.8％)。MS(ESI)M/Z:672.25[M+H] +。 It was quenched with water (5 mL), extracted with ethyl acetate (20 mL×2 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 mL), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. . The obtained residue was purified by silica gel column chromatography (n-hexane/ethyl acetate = 1:2) to obtain 24 mg of oily (S)-3-(4-(2-(2-(((5-chloro-2 -(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)phenyl)-5-oxo-2,5-dihydro- 1H-pyrazole-1-carboxylic acid tert-butyl ester (yield: 35.8%). MS (ESI) M/Z: 672.25 [M+H] + .
步骤E：合成(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((4-(5-氧代-2,5-二氢-1H-吡唑-3-基)苯基)氨基)-3-苯基丙-2-基)草酰胺 Step E: Synthesis of (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -(1-oxo-1-((4-(5- Oxo-2,5-dihydro-1H-pyrazol-3-yl)phenyl)amino)-3-phenylprop-2-yl)oxamide
Figure PCTCN2020133493-appb-000171
Figure PCTCN2020133493-appb-000171
室温下，将(S)-3-(4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯基)-5-氧代-2,5-二氢-1H-吡唑-1-羧酸叔丁酯(24毫克，0.036毫摩尔)溶于乙酸乙酯(2毫升)，向该溶液加入盐酸/乙酸乙酯溶液(1.0毫升，2摩尔/升)，室温反应过夜。At room temperature, (S)-3-(4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-phenylpropionamido)phenyl)-5-oxo-2,5-dihydro-1H-pyrazole-1-carboxylic acid tert-butyl ester (24 mg, 0.036 mmol) dissolved in acetic acid Ethyl acetate (2 mL), hydrochloric acid/ethyl acetate solution (1.0 mL, 2 mol/L) was added to the solution, and the reaction was carried out at room temperature overnight.
减压蒸除溶剂，残余物用制备HPLC纯化，得2.8毫克白色固体(S)-N 1-(5-氯-2-(1H-四唑-1-基)苯基)-N 2-(1-氧代-1-((4-(5-氧代-2,5-二氢-1H-吡唑-3-基)苯基)氨基)-3-苯基丙-2-基)草酰胺(收率13.6％)。MS(ESI)M/Z:570.10[M-H] -。 The solvent was evaporated under reduced pressure, and the residue was purified by preparative HPLC to obtain 2.8 mg of white solid (S)-N 1 -(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-N 2 -( 1-oxo-1-((4-(5-oxo-2,5-dihydro-1H-pyrazol-3-yl)phenyl)amino)-3-phenylprop-2-yl)grass Amide (yield 13.6%). MS(ESI) M/Z: 570.10 [MH] - .
实施例23Example 23
合成(S)-4-(2-(2-((5-氯-2-(三氟甲氧基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000172
Figure PCTCN2020133493-appb-000172
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成2-((5-氯-2-(三氟甲氧基)苯基)氨基)-2-氧代乙酸Step A: Synthesis of 2-((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000173
Figure PCTCN2020133493-appb-000173
室温下，向5-氯-2-(三氟甲氧基)苯胺(211毫克，1.0毫摩尔)的二氯甲烷(3毫升)溶液依次加入吡啶(119毫克，1.5毫摩尔)，氯草酸单甲酯(184毫克，1.5毫摩尔)，室温反应2小时。加水(5毫升)淬灭，用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，最后减压浓缩，所得粗产物直接用于下一步反应。At room temperature, to a solution of 5-chloro-2-(trifluoromethoxy)aniline (211 mg, 1.0 mmol) in dichloromethane (3 mL) was added pyridine (119 mg, 1.5 mmol) and chlorooxalic acid in turn. Methyl ester (184 mg, 1.5 mmol) was reacted at room temperature for 2 hours. It was quenched with water (5 mL), extracted with ethyl acetate (30 mL×2 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 mL), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. , The resulting crude product was directly used in the next reaction.
将上述粗产物溶于四氢呋喃/水(10毫升/5毫升)，室温反应2小时。减压蒸除有机溶剂，用2摩尔/升盐酸调节pH至1-2，析出固体，抽滤，滤饼用少量水洗涤3次，烘干得250毫克淡黄色固体2-((5-氯-2-(三氟甲氧基)苯基)氨基)-2-氧代乙酸(收率：88.3％)。MS(ESI)M/Z:281.97[M-H] -。 The above crude product was dissolved in tetrahydrofuran/water (10 mL/5 mL) and reacted at room temperature for 2 hours. The organic solvent was evaporated under reduced pressure, the pH was adjusted to 1-2 with 2 mol/L hydrochloric acid, the solid was precipitated, and the filter cake was washed with a small amount of water 3 times, and dried to obtain 250 mg of light yellow solid 2-((5-chloro -2-(Trifluoromethoxy)phenyl)amino)-2-oxoacetic acid (yield: 88.3%). MS(ESI) M/Z: 281.97 [MH] - .
步骤B：合成(S)-4-(2-(2-(((5-氯-2-(三氟甲氧基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000174
Figure PCTCN2020133493-appb-000174
室温下，将2-((5-氯-2-(三氟甲氧基)苯基)氨基)-2-氧代乙酸(85毫克，0.30毫摩尔)、(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(146毫克，0.30毫摩尔)和1-丙基磷酸酐(394毫克，0.60毫摩尔)加入N,N-二甲基甲酰胺(2.0毫升)中，滴加N,N-二异丙基乙胺(155毫克，1.20毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, 2-((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetic acid (85 mg, 0.30 mmol), (S)-4-(2- Amino-3-phenylpropionamido) tert-butyl benzoate (146 mg, 0.30 mmol) and 1-propyl phosphoric anhydride (394 mg, 0.60 mmol) were added to N,N-dimethylformamide (2.0 (Ml), N,N-diisopropylethylamine (155 mg, 1.20 mmol) was added dropwise, and after the dropping, the reaction was carried out at room temperature overnight under the protection of N 2.
反应结束，加水(5毫升)淬灭，用乙酸乙酯(30毫升×2次)萃取，合并有机相，有机相先用饱和食盐水(20毫升)洗涤，然后用无水硫酸钠干燥，最后减压浓缩，残余物用制备HPLC纯化，得14毫克无色油状物(S)-4-(2-(2-(((5-氯-2-(三氟甲氧基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：7.7％)。MS(ESI)M/Z:606.20[M+H] +。 After the reaction was completed, it was quenched by adding water (5 mL), extracted with ethyl acetate (30 mL×2 times), and the organic phases were combined. The organic phase was washed with saturated brine (20 mL), then dried with anhydrous sodium sulfate, and finally Concentrated under reduced pressure, and the residue was purified by preparative HPLC to obtain 14 mg of colorless oil (S)-4-(2-(2-(((5-chloro-2-(trifluoromethoxy)phenyl)amino )-2-oxoacetamido)-3-phenylpropionamido)tert-butyl benzoate (yield: 7.7%). MS (ESI) M/Z: 606.20 [M+H] + .
步骤C：合成(S)-4-(2-(2-((5-氯-2-(三氟甲氧基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetamido)-3-phenylpropane Amido) benzoic acid
Figure PCTCN2020133493-appb-000175
Figure PCTCN2020133493-appb-000175
室温下，向(S)-4-(2-(2-(((5-氯-2-(三氟甲氧基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(14毫克，0.023毫摩尔)的二氯甲烷(1.5毫升)溶液中加入三氟乙酸(0.4毫升)中，加毕，室温反应2小时。减压蒸干溶剂，得10毫克淡黄色固体(S)-4-(2-(2-((5-氯-2-(三氟甲氧基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：79.1％)。MS(ESI)M/Z:548.08[M-H] -。 At room temperature, to (S)-4-(2-(2-(((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetamido)-3-benzene Add trifluoroacetic acid (0.4 mL) to a solution of tert-butyl propionamido)benzoate (14 mg, 0.023 mmol) in dichloromethane (1.5 mL). After the addition is complete, react at room temperature for 2 hours. Evaporate to dryness under reduced pressure Solvent, get 10 mg of light yellow solid (S)-4-(2-(2-((5-chloro-2-(trifluoromethoxy)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)benzoic acid (yield: 79.1%). MS (ESI) M/Z: 548.08 [MH] - .
实施例24Example 24
合成(S)-4-(2-(2-(((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000176
Figure PCTCN2020133493-appb-000176
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成1-(4-氯-2-硝基苯基)-1H-1,2,4-三唑Step A: Synthesis of 1-(4-chloro-2-nitrophenyl)-1H-1,2,4-triazole
Figure PCTCN2020133493-appb-000177
Figure PCTCN2020133493-appb-000177
室温下，将5-氯-2-氟硝基苯(1.0克，5.7毫摩尔)、1,2,4-三氮唑(462.5毫克，6.7毫摩尔)和碳酸钾(945.4毫克，6.8毫摩尔)加入N,N-二甲基甲酰胺(10.0毫升)中，N 2保护下，80摄氏度反应1.5小时。 At room temperature, 5-chloro-2-fluoronitrobenzene (1.0 g, 5.7 mmol), 1,2,4-triazole (462.5 mg, 6.7 mmol) and potassium carbonate (945.4 mg, 6.8 mmol) ) Was added to N,N-dimethylformamide (10.0 ml), and reacted at 80 degrees Celsius for 1.5 hours under the protection of N 2.
反应结束，垫无水硫酸钠抽滤，滤饼用50.0毫升乙酸乙酯洗涤，滤液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(50毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩得到1.1克棕色油状物1-(4-氯-2-硝基苯基)-1H-1,2,4-三唑，无需纯化，直接用于下一步反应。LCMS:RT＝3.28min,[M+H] +＝225.06。 After the reaction was over, a pad of anhydrous sodium sulfate was suction filtered, the filter cake was washed with 50.0 ml ethyl acetate, the filtrate was extracted with ethyl acetate (30 ml × 3 times), the organic phases were combined, and the organic phase was first washed with saturated brine (50 ml × 2 times) washing, then drying with anhydrous sodium sulfate, and finally concentrating under reduced pressure to obtain 1.1 g of brown oil 1-(4-chloro-2-nitrophenyl)-1H-1,2,4-triazole. Purified and used directly in the next reaction. LCMS: RT = 3.28 min, [M+H] + =225.06.
步骤B：合成5-氯-2-(1H-1,2,4-三唑-1-基)苯胺Step B: Synthesis of 5-chloro-2-(1H-1,2,4-triazol-1-yl)aniline
Figure PCTCN2020133493-appb-000178
Figure PCTCN2020133493-appb-000178
室温下，将1-(4-氯-2-硝基苯基)-1H-1,2,4-三唑(1.1克，4.9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(11.1克，49.0毫摩尔)，N 2保护下，室温反应2小时。 At room temperature, add 1-(4-chloro-2-nitrophenyl)-1H-1,2,4-triazole (1.1 g, 4.9 mmol) into ethyl acetate, add in batches under ice bath Stannous chloride dihydrate (11.1 g, 49.0 mmol) was reacted at room temperature for 2 hours under the protection of N 2.
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50毫升乙酸乙酯洗涤，滤液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(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：纯正己烷)。得到820.0毫克乳白色固体5-氯-2-(1H-1,2,4-三唑-1-基)苯胺(收率：86.2％)。LCMS:RT＝3.07min,[M+H] +＝195.09。 After the reaction was completed, it was quenched with saturated sodium bicarbonate aqueous solution, and excess sodium bicarbonate was added to adjust the pH to weakly alkaline. The filter cake was filtered with Celite pad and the filter cake was washed with 50 ml of ethyl acetate. The filtrate was washed with ethyl acetate ( 30 ml×3 times), the organic phases were combined, and the organic phase was washed with saturated brine (50 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). Obtained 820.0 mg of creamy white solid 5-chloro-2-(1H-1,2,4-triazol-1-yl)aniline (yield: 86.2%). LCMS: RT=3.07min, [M+H] + =195.09.
步骤C：合成2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000179
Figure PCTCN2020133493-appb-000179
N 2保护、冰浴下，向含有5-氯-2-(1H-1,2,4-三唑-1-基)苯胺(820.0毫克，4.2毫摩尔)和吡啶(332.2毫克，4.2毫摩尔)的二氯甲烷(30毫升)中滴加草酰氯单甲酯(624.7毫克，5.1毫摩尔)，滴毕，室温反应30分钟。 Under the protection of N 2 and under ice bath, it contains 5-chloro-2-(1H-1,2,4-triazol-1-yl)aniline (820.0 mg, 4.2 mmol) and pyridine (332.2 mg, 4.2 mmol) ) Was added dropwise to dichloromethane (30 ml) of oxalyl chloride monomethyl ester (624.7 mg, 5.1 mmol), after dropping, 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(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×3次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩得到1.1克白色固体2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸甲酯，无需纯化，直接用于下一步反应。LCMS:RT＝3.29min,[M-H] -＝281.09。 After the reaction was over, it was quenched by adding water, and the mixture was extracted with dichloromethane (50 ml×3 times). The organic phases were combined, and the organic phase was washed with saturated brine (30 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure to obtain 1.1 g of white solid 2-((5-chloro-2-(1H- Methyl 1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetate was used directly in the next reaction without purification. LCMS: RT = 3.29 min, [MH] - =281.09.
步骤D：合成2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸Step D: Synthesis of 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000180
Figure PCTCN2020133493-appb-000180
冰浴下，向含有2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸甲酯(1.1克，3.9毫摩尔)的四氢呋喃(100毫升)中滴加氢氧化锂(3.2克，7.8毫摩尔)水溶液(50毫升)，滴毕即反应结束。Under ice bath, add methyl 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetate (1.1 g, 3.9 Aqueous solution (50 ml) of lithium hydroxide (3.2 g, 7.8 mmol) was added dropwise to tetrahydrofuran (100 ml) in millimoles), 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(30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，最后减压浓缩得到810.5毫克淡黄色固体2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸，无需纯化，直接用于下一步反应。LCMS:RT＝1.76min,[M-H] -＝265.01。 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 (30 ml×3 times), combine the organic phases, and use saturated brine (30 Ml×2 times), then dried with anhydrous sodium sulfate, and finally concentrated under reduced pressure to obtain 810.5 mg of pale yellow solid 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl )Phenyl)amino)-2-oxoacetic acid, without purification, directly used in the next reaction. LCMS: RT=1.76 min, [MH] - =265.01.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxy (Acetamido)-3-phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000181
Figure PCTCN2020133493-appb-000181
室温下，将2-((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酸(500.0毫克，1.9毫摩尔)、(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(646.0毫克，1.9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(1.1克，2.9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(736.7毫克，5.7毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 2-((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxoacetic acid (500.0 mg, 1.9 mmol), (S)-4-(2-Amino-3-phenylpropionamido) tert-butyl benzoate (646.0 mg, 1.9 mmol) and 2-(7-benzotriazole oxide)-N,N, N',N'-tetramethylurea hexafluorophosphate (1.1g, 2.9mmol) was added to N,N-dimethylformamide (5.0ml), and N,N-diisopropylethylamine was added dropwise (736.7 mg, 5.7 mmol), after dripping, under N 2 protection, react at room temperature for 5 hours.
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＝2/1)。得到458.2毫克黄色固体(S)-4-(2-(2-(((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：40.9％)。LCMS:RT＝4.33min,[M-H] -＝587.21。 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/n-hexane=2/1). Obtained 458.2 mg of yellow solid (S)-4-(2-(2-(((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2- Oxoacetamido)-3-phenylpropionamido) tert-butyl benzoate (yield: 40.9%). LCMS: RT = 4.33 min, [MH] - =587.21.
步骤F：合成(S)-4-(2-(2-(((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxy (Acetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000182
Figure PCTCN2020133493-appb-000182
室温下，将(S)-4-(2-(2-(((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(60.0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2小时。At room temperature, the (S)-4-(2-(2-(((5-chloro-2-(1H-1,2,4-triazol-1-yl)phenyl)amino)-2-oxy Acetamido)-3-phenylpropionamido) tert-butyl benzoate (60.0 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 2 hour.
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35.1毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-1,2,4-三唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：65.9％)。LCMS:RT＝3.67min,[M-H] -＝531.06。 1H NMR(500MHz,DMSO)δ12.75(s,1H),11.17(s,1H),10.43(s,1H),9.17(d,J＝8.2Hz,1H),9.09(s,1H),8.38(s,1H),8.36(d,J＝2.3Hz,1H),7.91(d,J＝8.7Hz,2H),7.78(d,J＝8.6Hz,1H),7.70(d,J＝8.7Hz,2H),7.49(dd,J＝8.6,2.4Hz,1H),7.28(dt,J＝15.2,7.5Hz,4H),7.23–7.16(m,1H),4.79–4.69(m,1H),3.25–3.13(m,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 35.1 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-1,2,4-triazol-1-yl) Phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid (yield: 65.9%). LCMS: RT = 3.67 min, [MH] - = 531.06. 1 H NMR (500MHz,DMSO)δ12.75(s,1H),11.17(s,1H),10.43(s,1H),9.17(d,J=8.2Hz,1H),9.09(s,1H),8.38(s ,1H),8.36(d,J=2.3Hz,1H),7.91(d,J=8.7Hz,2H),7.78(d,J=8.6Hz,1H),7.70(d,J=8.7Hz,2H ),7.49(dd,J=8.6,2.4Hz,1H), 7.28(dt,J=15.2,7.5Hz,4H), 7.23–7.16(m,1H), 4.79–4.69(m,1H), 3.25– 3.13 (m, 2H).
实施例25Example 25
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000183
Figure PCTCN2020133493-appb-000183
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000184
Figure PCTCN2020133493-appb-000184
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(120.0毫克，0.16毫摩尔)和哌啶(136.2毫克，1.6毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(206.8毫克，1.6毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (120.0 mg, 0.16 mmol) and piperidine (136.2 mg, 1.6 mmol) N,N-diisopropylethylamine (206.8 mg, 1.6 mmol) was added dropwise to tetrahydrofuran (3.0 ml) in mol). 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：乙酸乙酯/正己烷＝1/2)。得到62毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：51.3％)。LCMS:RT＝4.11min,[M-H] -＝753.35。 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 62 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 51.3%). LCMS: RT = 4.11 min, [MH] - =753.35.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000185
Figure PCTCN2020133493-appb-000185
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(62.0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 (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (62.0 mg, 0.08 mmol) was added to dichloromethane (2.0 mL) In, 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(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：50.1％)。LCMS:RT＝3.53min,[M+H] +＝699.22。 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 28 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(piperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 50.1%). LCMS: RT = 3.53 min, [M+H] + =699.22.
实施例26Example 26
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基))-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧咪二氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino))-2-oxoacetamido)-3- (4-(4-Cyanopiperidine-1-carboximiddiamino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000186
Figure PCTCN2020133493-appb-000186
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(3-(4-(1H-咪唑-1-羧酰亚胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(3-(4-(1H-imidazole-1-carboximido)phenyl)-2-(2-((5-chloro-2-(1H-tetra (Azol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000187
Figure PCTCN2020133493-appb-000187
室温下，将(S)-5-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(500.0毫克，0.78毫摩尔)和二(1H-咪唑基)甲亚胺(388.0毫克，2.4毫摩尔)加入到N,N-二甲基甲酰胺(10.0毫升)中，90摄氏度反应5小时。At room temperature, (S)-5-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido) propionamido) -1H-indole-2-carboxylic acid tert-butyl ester (500.0 mg, 0.78 mmol) and bis(1H-imidazolyl) azomethine (388.0 mg, 2.4 mmol) Mol) was added to N,N-dimethylformamide (10.0 ml), and reacted at 90 degrees Celsius for 5 hours.
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232毫克白色固体(S)-5-(3-(4-(1H-咪唑-1-羧酰亚胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：40.4％)。LCMS:RT＝3.61min,[M+H] +＝737.29。 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: dichloromethane/methanol=10/1). Obtain 232 mg of white solid (S)-5-(3-(4-(1H-imidazole-1-carboximido)phenyl)-2-(2-((5-chloro-2-(1H- Tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 40.4%). LCMS: RT = 3.61 min, [M+H] + =737.29.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧咪二胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step B: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Cyanopiperidine-1-carboximidamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000188
Figure PCTCN2020133493-appb-000188
室温下，将(S)-5-(3-(4-(1H-咪唑-1-羧酰亚胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(120.0毫克，0.16毫摩尔)和4-氰基哌啶(53.8毫克，0.48毫摩尔)加入四氢呋喃(3.0毫升)中，65摄氏度反应过夜。At room temperature, (S)-5-(3-(4-(1H-imidazole-1-carboximido)phenyl)-2-(2-((5-chloro-2-(1H-tetra (Azol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (120.0 mg, 0.16 mmol) and 4-cyanopiper Pyridine (53.8 mg, 0.48 mmol) was added to tetrahydrofuran (3.0 mL) and reacted 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54毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧咪二胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：43.3％)。LCMS:RT＝3.82min,[M-H] -＝777.27。 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=1/1). Obtain 54 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-Cyanopiperidine-1-carboximidamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 43.3%). LCMS: RT = 3.82 min, [MH] - =777.27.
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基))-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧咪二氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino))-2-oxoacetamido) -3-(4-(4-Cyanopiperidine-1-carboximidylamino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000189
Figure PCTCN2020133493-appb-000189
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧咪二胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(54.0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, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Cyanopiperidine-1-carboximidamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (54.0 mg, 0.07 mmol) was added with dichloride Trifluoroacetic acid (0.5 mL) was added dropwise to methane (2.0 mL), 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17毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基))-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧咪二氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：50.1％)。LCMS:RT＝3.53min,[M-H] -＝722.32。 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 17 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)) -2-oxoacetamido)-3-(4-(4-cyanopiperidine-1-carboximiddiamino)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield : 50.1%). LCMS: RT = 3.53 min, [MH] - = 722.32.
实施例27Example 27
合成(S,Z)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(N',4-二氰基哌啶-1-羧咪二氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3- (4-(N',4-dicyanopiperidine-1-carboximidodiamino)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000190
Figure PCTCN2020133493-appb-000190
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S,Z)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(((氰基氨基)(苯氧基)甲基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(((cyanoamino)(phenoxy)methyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000191
Figure PCTCN2020133493-appb-000191
室温下，将(S)-5-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(120.0毫克，0.19毫摩尔)和N-氰基羰亚胺二苯基酯(135.7毫克，0.57毫摩尔)加入到异丙醇(5.0毫升)中，60摄氏度反应过夜。At room temperature, (S)-5-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido) propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (120.0 mg, 0.19 mmol) and N-cyanocarbonylimide diphenyl ester (135.7 mg, 0.57 (Mmol) was added to isopropanol (5.0 ml) and reacted 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：乙酸乙酯/正己烷＝2/1)。得到70毫克白色固体(S,Z)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(((氰基氨基)(苯氧基)甲基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：46.7％)。LCMS:RT＝4.11min,[M+H] +＝788.28。 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=2/1). Obtain 70 mg of white solid (S, Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(((cyanoamino)(phenoxy)methyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (Yield: 46.7% ). LCMS: RT = 4.11 min, [M+H] + =788.28.
步骤B：合成(S,Z)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(N',4-二氰基哌啶-1-羧咪二氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step B: Synthesis of (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(N',4-dicyanopiperidine-1-carboximidodiamino)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000192
Figure PCTCN2020133493-appb-000192
室温下，向含有(S,Z)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(((氰基氨基)(苯氧基)甲基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(70.0毫克，0.09毫摩尔)和4-氰基哌啶(19.8毫克，0.18毫摩尔)的乙腈(3.0毫升)中滴加三乙胺(9.2毫克，0.09毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(((cyanoamino)(phenoxy)methyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (70.0 mg, 0.09 mg Triethylamine (9.2 mg, 0.09 mmol) was added dropwise to acetonitrile (3.0 ml) with 4-cyanopiperidine (19.8 mg, 0.18 mmol) and the reaction was completed 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：乙酸乙酯/正己烷＝1/2)。得到58毫克白色固体(S,Z)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(N',4-二氰基哌啶-1-羧咪二氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：80.1％)。LCMS:RT＝3.90min,[M+H] +＝804.28。 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 = 1/2). Obtain 58 mg of white solid (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(N',4-dicyanopiperidine-1-carboximidodiamino)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 80.1 %). LCMS: RT = 3.90 min, [M+H] + =804.28.
步骤C：合成(S,Z)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(N',4-二氰基哌啶-1-羧咪二氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step C: Synthesis of (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(N',4-dicyanopiperidine-1-carboximiddiamino)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000193
Figure PCTCN2020133493-appb-000193
室温下，将(S,Z)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(N',4-二氰基哌啶-1-羧咪二氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(58.0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, the (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(N',4-dicyanopiperidine-1-carboximidodiamino)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (58.0 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.
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23毫克白色固体(S,Z)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(N',4-二氰基哌啶-1-羧咪二氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：44.0％)。LCMS:RT＝2.83min,[M+H] +＝748.19。 1H NMR(400MHz,DMSO)δ12.95(s,1H),11.73(s,1H),10.72(s,1H),10.68(s,1H),10.07(s,1H),9.86(s,1H),9.83(s,1H),8.90(d,J＝8.4Hz,1H),7.96(d,J＝2.3Hz,1H),7.94(d,J＝1.6Hz,1H),7.78(d,J＝8.6Hz,1H),7.63(dd,J＝8.6,2.4Hz,1H),7.38(d,J＝9.3Hz,1H),7.36–7.29(m,2H),7.09(d,J＝8.3Hz,2H),7.06(d,J＝2.0Hz,1H),4.77–4.66(m,1H),3.82–3.55(m,4H),3.17(d,J＝7.9Hz,2H),2.11–1.83(m,5H)。 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 23 mg of white solid (S,Z)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino) -2-oxoacetamido)-3-(4-(N',4-dicyanopiperidine-1-carboximiddiamino)phenylpropionamido)-1H-indole-2-carboxylic acid (Yield: 44.0%). LCMS: RT = 2.83 min, [M+H] + = 748.19. 1 H NMR (400MHz, DMSO) δ 12.95 (s, 1H), 11.73 (s, 1H), 10.72 ( s, 1H), 10.68 (s, 1H), 10.07 (s, 1H), 9.86 (s, 1H), 9.83 (s, 1H), 8.90 (d, J = 8.4 Hz, 1H), 7.96 (d, J ＝2.3Hz,1H),7.94(d,J＝1.6Hz,1H),7.78(d,J＝8.6Hz,1H),7.63(dd,J＝8.6,2.4Hz,1H),7.38(d,J =9.3Hz,1H),7.36–7.29(m,2H), 7.09(d,J=8.3Hz,2H), 7.06(d,J=2.0Hz,1H),4.77–4.66(m,1H),3.82 -3.55 (m, 4H), 3.17 (d, J = 7.9 Hz, 2H), 2.11-1.83 (m, 5H).
实施例28Example 28
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-苯基脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-phenylureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000194
Figure PCTCN2020133493-appb-000194
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成 异氰酸苯酯Step A: Synthesis of phenyl isocyanate
Figure PCTCN2020133493-appb-000195
Figure PCTCN2020133493-appb-000195
冰浴下，向含有苯胺(50.0毫克，0.54毫摩尔)的乙酸乙酯(5.0毫升)中滴加三光气(240.37毫克，0.81毫摩尔)，滴毕，80摄氏度反应4小时。Under an ice bath, triphosgene (240.37 mg, 0.81 mmol) was added dropwise to ethyl acetate (5.0 ml) containing aniline (50.0 mg, 0.54 mmol), the dripping was completed, and the reaction was carried out at 80 degrees Celsius for 4 hours.
反应结束，减压浓缩得到58毫克棕色油状物异氰酸苯酯，无需纯化，直接用于下一步反应。After the reaction was completed, it was concentrated under reduced pressure to obtain 58 mg of brown oily phenyl isocyanate, which was directly used in the next reaction without purification.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-苯基脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step B: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-phenylureido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000196
Figure PCTCN2020133493-appb-000196
冰浴下，将异氰酸苯酯(32.0毫克，0.27毫摩尔)的四氢呋喃(1.0毫升)滴加入(S)-5-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(58.0毫克，0.09毫摩尔)和三乙胺(36.5毫克，0.36毫摩尔)的四氢呋喃(2.0毫升)中，室温反应过夜。Under an ice bath, phenyl isocyanate (32.0 mg, 0.27 mmol) and tetrahydrofuran (1.0 ml) were added dropwise to (S)-5-(3-(4-aminophenyl)-2-(2-(( 5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (58.0 mg , 0.09 mmol) and triethylamine (36.5 mg, 0.36 mmol) in tetrahydrofuran (2.0 mL) at room temperature 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＝2/1)。得到47毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-苯基脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：68.5％)。LCMS:RT＝4.14min,[M+H] +＝763.27。 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 = 2/1). Obtain 47 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-phenylureido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 68.5%). LCMS: RT = 4.14 min, [M+H] + =763.27.
步骤C：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-苯基脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step C: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-phenylureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000197
Figure PCTCN2020133493-appb-000197
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-苯基脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(47.0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 (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-phenylureido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (47.0 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18毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-苯基脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：42.4％)。LCMS:RT＝3.72min,[M-H] -＝705.15。 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 (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(3-phenylureido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 42.4%). LCMS: RT = 3.72 min, [MH] - =705.15.
实施例29Example 29
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-氧吡咯烷-1-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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(2-oxopyrrolidin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000198
Figure PCTCN2020133493-appb-000198
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成1-(1-苄基哌啶-4-基)吡咯烷-2-酮Step A: Synthesis of 1-(1-benzylpiperidin-4-yl)pyrrolidin-2-one
Figure PCTCN2020133493-appb-000199
Figure PCTCN2020133493-appb-000199
室温下，将4-氨基-1-苄基哌啶(3.0克，15.8毫摩尔)、4-溴丁酸乙酯(3.1克，15.8毫摩尔)和碳酸钾(2.6克，19.0毫摩尔)加入N,N-二甲基甲酰胺(20.0毫升)中，N 2保护下，室温反应过夜。 At room temperature, 4-amino-1-benzylpiperidine (3.0 g, 15.8 mmol), ethyl 4-bromobutyrate (3.1 g, 15.8 mmol) and potassium carbonate (2.6 g, 19.0 mmol) were added In N,N-dimethylformamide (20.0 mL), under the protection of N 2 , the reaction was carried out at room temperature overnight.
反应结束，垫无水硫酸钠抽滤，滤饼用100.0毫升二氯甲烷洗涤，滤液用二氯甲烷(50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：二氯甲烷/甲醇＝4/1)。得到1.9克类白色油状物1-(1-苄基哌啶-4-基)吡咯烷-2-酮(收率：46.5％)。LCMS:RT＝0.93min,[M+H] +＝259.22。 After the reaction, the pad was suction filtered with anhydrous sodium sulfate, the filter cake was washed with 100.0 ml of dichloromethane, the filtrate was extracted with dichloromethane (50 ml × 3 times), the organic phases were combined, and the organic phase was first used with saturated brine (50 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: dichloromethane/methanol=4/1). 1.9 grams of white oily 1-(1-benzylpiperidin-4-yl)pyrrolidin-2-one was obtained (yield: 46.5%). LCMS: RT=0.93 min, [M+H] + =259.22.
步骤B：合成1-(哌啶-4-基)吡咯烷-2-酮Step B: Synthesis of 1-(piperidin-4-yl)pyrrolidin-2-one
Figure PCTCN2020133493-appb-000200
Figure PCTCN2020133493-appb-000200
室温下，将1-(1-苄基哌啶-4-基)吡咯烷-2-酮(200.0毫克，0.77毫摩尔)溶于甲醇(5.0毫升)中，加入钯碳(20.0毫克)，安装氢气球，室温反应10小时。At room temperature, dissolve 1-(1-benzylpiperidin-4-yl)pyrrolidin-2-one (200.0 mg, 0.77 mmol) in methanol (5.0 mL), add palladium on carbon (20.0 mg), and install Hydrogen balloon, react at room temperature for 10 hours.
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：二氯甲烷/甲醇＝10/1)。得到100毫克棕色固体1-(哌啶-4-基)吡咯烷-2-酮(收率：77.2％)。LCMS:RT＝0.65min,[M+H] +＝169.13。 After the reaction was completed, 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: dichloromethane/methanol=10/1). 100 mg of brown solid 1-(piperidin-4-yl)pyrrolidin-2-one was obtained (yield: 77.2%). LCMS: RT=0.65 min, [M+H] + =169.13.
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-氧吡咯烷-1-基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-oxopyrrolidin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000201
Figure PCTCN2020133493-appb-000201
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(120.0毫克，0.16毫摩尔)和1-(哌啶-4-基)吡咯烷-2-酮(80.7毫克，0.48毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(82.7毫克，0.64毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (120.0 mg, 0.16 mmol) and 1-(piperidine-4- N,N-diisopropylethylamine (82.7 mg, 0.64 mmol) was added dropwise to tetrahydrofuran (3.0 ml) in pyrrolidin-2-one (80.7 mg, 0.48 mmol), 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＝1/1)。得到95毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-氧吡咯烷-1-基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：70.8％)。LCMS:RT＝3.82min,[M-H] -＝836.27。 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=1/1). Obtain 95 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(4-(2-oxopyrrolidin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester Rate: 70.8%). LCMS: RT = 3.82 min, [MH] - =836.27.
步骤D：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-氧吡咯烷-1-基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step D: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-oxopyrrolidin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000202
Figure PCTCN2020133493-appb-000202
室温下，将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-氧吡咯烷-1-基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(95.0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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-oxopyrrolidin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (95.0 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.
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-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-氧吡咯烷-1-基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：40.7％)。LCMS:RT＝3.36min,[M-H] -＝780.19。 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-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido)-3-(4-(4-(2-oxopyrrolidin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2 -Carboxylic acid (yield: 40.7%). LCMS: RT = 3.36 min, [MH] - = 780.19.
实施例30Example 30
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1,1-二氧化四氢-2H-硫代吡喃-4-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000203
Figure PCTCN2020133493-appb-000203
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1,1-二氧化四氢-2H-硫代吡喃-4-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000204
Figure PCTCN2020133493-appb-000204
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(120.0毫克，0.16毫摩尔)和4-氨基四氢-2H-噻喃-1,1-二氧化物盐酸盐(89.1毫克，0.48毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(124.1毫克，0.96毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (120.0 mg, 0.16 mmol) and 4-aminotetrahydro-2H- Thian-1,1-dioxide hydrochloride (89.1 mg, 0.48 mmol) in tetrahydrofuran (3.0 ml) was added dropwise with N,N-diisopropylethylamine (124.1 mg, 0.96 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61毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1,1-二氧化四氢-2H-硫代吡喃-4-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：46.5％)。LCMS:RT＝4.24min,[M-H] -＝817.24。 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 = 1/2). Obtain 61 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid tert Butyl ester (yield: 46.5%). LCMS: RT = 4.24 min, [MH] - =817.24.
步骤B：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1,1-二氧化四氢-2H-硫代吡喃-4-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000205
Figure PCTCN2020133493-appb-000205
室温下，将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1,1-二氧化四氢-2H-硫代吡喃-4-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(61.0毫克，0.075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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl The ester (61.0 mg, 0.075 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.
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(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1,1-二氧化四氢-2H-硫代吡喃-4-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：41.9％)。LCMS:RT＝3.30min,[M-H] -＝761.07。 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 (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido)-3-(4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)ureido)phenylpropionamido)-1H- Indole-2-carboxylic acid (yield: 41.9%). LCMS: RT = 3.30 min, [MH] - =761.07.
实施例31Example 31
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(Methylsulfonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000206
Figure PCTCN2020133493-appb-000206
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000207
Figure PCTCN2020133493-appb-000207
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(120.0毫克，0.16毫摩尔)和4-甲磺酰基哌啶(78.4毫克，0.48毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(124.1毫克，0.96毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (120.0 mg, 0.16 mmol) and 4-methylsulfonylpiperidine ( 78.4 mg, 0.48 mmol) of tetrahydrofuran (3.0 mL) was added dropwise with N,N-diisopropylethylamine (124.1 mg, 0.96 mmol), 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＝1/2)。得到81毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-羧酸叔丁酯(收率：60.8％)。LCMS:RT＝4.27min,[M-H] -＝831.27。 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). Obtained 81 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 60.8%). LCMS: RT = 4.27 min, [MH] - =831.27.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000208
Figure PCTCN2020133493-appb-000208
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-羧酸叔丁酯(81.0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, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Methylsulfonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (81.0 mg, 0.10 mmol) was added Trifluoroacetic acid (0.5 mL) was added dropwise to dichloromethane (2.0 mL), 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毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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：55.3％)。LCMS:RT＝3.38min,[M-H] -＝774.98。 1H NMR(400MHz,DMSO)δ12.92(s,1H),11.73(s,1H),10.74(s,1H),10.05(s,1H),9.85(s,1H),8.78(d,J＝8.4Hz,1H),8.53(s,1H),8.00(d,J＝2.3Hz,1H),7.95(s,1H),7.78(d,J＝8.6Hz,1H),7.62(dd,J＝8.6,2.4Hz,1H),7.34(ddd,J＝10.7,10.2,5.4Hz,4H),7.14(d,J＝8.5Hz,2H),7.07(d,J＝1.5Hz,1H),4.73–4.63(m,1H),4.25(d,J＝13.3Hz,2H),3.08(d,J＝7.8Hz,2H),2.95(s,3H),2.83(t,J＝12.3Hz,2H),2.03(d,J＝11.2Hz,2H),1.62–1.44(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 43 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(4-(methylsulfonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid (yield : 55.3%). LCMS: RT = 3.38 min, [MH] - = 774.98. 1 H NMR (400MHz, DMSO) δ 12.92 (s, 1H), 11.73 (s, 1H), 10.74 (s, 1H), 10.05(s,1H), 9.85(s,1H), 8.78(d,J=8.4Hz,1H), 8.53(s,1H),8.00(d,J=2.3Hz,1H),7.95(s,1H) ), 7.78 (d, J = 8.6 Hz, 1H), 7.62 (dd, J = 8.6, 2.4 Hz, 1H), 7.34 (ddd, J = 10.7, 10.2, 5.4 Hz, 4H), 7.14 (d, J = 8.5Hz, 2H), 7.07 (d, J = 1.5 Hz, 1H), 4.73-4.63 (m, 1H), 4.25 (d, J = 13.3 Hz, 2H), 3.08 (d, J = 7.8 Hz, 2H) , 2.95 (s, 3H), 2.83 (t, J = 12.3 Hz, 2H), 2.03 (d, J = 11.2 Hz, 2H), 1.62-1.44 (m, 2H).
实施例32Example 32
合成5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000209
Figure PCTCN2020133493-appb-000209
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]癸烷-3-醇三氟乙酸盐Step A: Synthesis of 1-oxa-8-azaspiro[4.5]decane-3-ol trifluoroacetate
Figure PCTCN2020133493-appb-000210
Figure PCTCN2020133493-appb-000210
室温下，将N-叔丁氧羰基-1-氧杂-8-氮杂螺[4.5]癸烷-3-醇(100毫克，0.39毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应2小时。At room temperature, add N-tert-butoxycarbonyl-1-oxa-8-azaspiro[4.5]decan-3-ol (100 mg, 0.39 mmol) to dichloromethane (2.0 mL) and add dropwise Trifluoroacetic acid (1.0 mL) was reacted at room temperature for 2 hours.
反应结束，蒸干二氯甲烷并用油泵抽干三氟乙酸得到79毫克黄色油状物1-氧杂-8-氮杂螺[4.5]癸烷-3-醇三氟乙酸盐，无需纯化，直接用于下一步反应。LCMS:RT＝0.62min,[M-H] -＝158.13。 After the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump to obtain 79 mg of yellow oil, 1-oxa-8-azaspiro[4.5]decane-3-ol trifluoroacetate, without purification, directly Used in the next reaction. LCMS: RT=0.62 min, [MH] - =158.13.
步骤B：合成5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step B: Synthesis of 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000211
Figure PCTCN2020133493-appb-000211
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(50.0毫克，0.065毫摩尔)和1-氧杂-8-氮杂螺[4.5]癸烷-3-醇三氟乙酸盐(66.3毫克，0.26毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(84.0毫克，0.65毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (50.0 mg, 0.065 mmol) and 1-oxa-8-nitrogen Heterspiro[4.5]decane-3-ol trifluoroacetate (66.3 mg, 0.26 mmol) in tetrahydrofuran (3.0 mL) was added dropwise N,N-diisopropylethylamine (84.0 mg, 0.65 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＝1/1)。得到46毫克白色固体5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：85.5％)。LCMS:RT＝4.23min,[M-H] -＝825.37。 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=1/1). Obtain 46 mg of white solid 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert Butyl ester (yield: 85.5%). LCMS: RT = 4.23 min, [MH] - =825.37.
步骤C：合成5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step C: Synthesis of 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000212
Figure PCTCN2020133493-appb-000212
室温下，将5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(46.0毫克，0.056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-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-Hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl The ester (46.0 mg, 0.056 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.
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-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基-1-氧杂-8-氮杂螺[4.5]癸烷-8-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：41.7％)。LCMS:RT＝3.32min,[M+H] +＝771.17。 1H NMR(400MHz,DMSO)δ11.72(s,1H),10.73(s,1H),10.04(s,1H),9.84(s,1H),8.76(d,J＝8.2Hz,1H),8.41(s,1H),7.99(d,J＝2.3Hz,1H),7.94(d,J＝1.1Hz,1H),7.77(d,J＝8.6Hz,1H),7.61(dd,J＝8.6,2.4Hz,1H),7.43–7.28(m,4H),7.11(d,J＝8.7Hz,2H),7.06(d,J＝1.6Hz,1H),4.71–4.63(m,1H),4.38–4.27(m,1H),3.81(dd,J＝9.1,4.7Hz,1H),3.62–3.50(m,2H),3.40–3.28(m,2H),3.07(d,J＝7.0Hz,2H),2.08–1.95(m,1H),1.87(dd,J＝13.2,6.7Hz,1H),1.78–1.56(m,3H),1.49(t,J＝5.8Hz,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 18 mg of white solid 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(3-hydroxy-1-oxa-8-azaspiro[4.5]decane-8-carboxamido)phenyl)propionamido)-1H- Indole-2-carboxylic acid (yield: 41.7%). LCMS: RT = 3.32 min, [M+H] + =771.17. 1 H NMR (400MHz, DMSO) δ 11.72 (s, 1H), 10.73 (s, 1H), 10.04 (s, 1H), 9.84 (s, 1H), 8.76 (d, J = 8.2 Hz, 1H), 8.41 (s, 1H), 7.99 (d, J = 2.3 Hz, 1H), 7.94 (d, J = 1.1 Hz, 1H), 7.77 (d, J = 8.6 Hz, 1H), 7.61 (dd, J = 8.6 ,2.4Hz,1H),7.43-7.28(m,4H),7.11(d,J=8.7Hz,2H),7.06(d,J=1.6Hz,1H),4.71-4.63(m,1H),4.38 –4.27(m,1H),3.81(dd,J=9.1,4.7Hz,1H),3.62–3.50(m,2H),3.40–3.28(m,2H),3.07(d,J=7.0Hz,2H ), 2.08–1.95 (m, 1H), 1.87 (dd, J = 13.2, 6.7 Hz, 1H), 1.78–1.56 (m, 3H), 1.49 (t, J = 5.8 Hz, 2H).
实施例33Example 33
合成(S)-5-(3-(4-(4-氨基甲酰基哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(3-(4-(4-carbamoylpiperidine-1-carboxamido)phenyl)-2-(2-((5-chloro-2-(1H-tetrazole) -1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000213
Figure PCTCN2020133493-appb-000213
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-((甲基磺酰基)氨基甲酰基)哌啶-1-甲酸叔丁酯Step A: Synthesis of tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000214
Figure PCTCN2020133493-appb-000214
室温下，将1-(叔丁氧羰基)哌啶-4-羧酸(1.0克，4.4毫摩尔)、甲磺酰胺(376.2毫克，4.0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(2.3克，6.0毫摩尔)加入N,N-二甲基甲酰胺(10.0毫升)中，滴加N,N-二异丙基乙胺(1.5克，11.8毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (1.0 g, 4.4 mmol), methanesulfonamide (376.2 mg, 4.0 mmol) and 2-(7-oxybenzotriazide) Azole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.3g, 6.0mmol) was added to N,N-dimethylformamide (10.0ml), and N,N was added dropwise -Diisopropylethylamine (1.5 g, 11.8 mmol), after dripping , react under the protection of N 2 at room temperature for 5 hours.
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(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：二氯甲烷/甲醇＝10/1)。得到700毫克黄色固体4-((甲基磺酰基)氨基甲酰基)哌啶-1-甲酸叔丁酯(收率：57.1％)。LCMS:RT＝3.36min,[M-H] -＝305.08。 After the reaction was over, 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 (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: dichloromethane/methanol=10/1). 700 mg of tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate was obtained as a yellow solid (yield: 57.1%). LCMS: RT = 3.36 min, [MH] - =305.08.
步骤B：合成N-(甲基磺酰基)哌啶-4-羧酰胺三氟乙酸盐Step B: Synthesis of N-(methylsulfonyl)piperidine-4-carboxamide trifluoroacetate
Figure PCTCN2020133493-appb-000215
Figure PCTCN2020133493-appb-000215
室温下，将4-((甲基磺酰基)氨基甲酰基)哌啶-1-甲酸叔丁酯(700毫克，2.3毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。At room temperature, tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate (700 mg, 2.3 mmol) was added to dichloromethane (2.0 mL), and trifluoroacetic acid ( 1.0 ml), 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得到240毫克黄色油状物N-(甲基磺酰基)哌啶-4-羧酰胺三氟乙酸盐，无需纯化，直接用于下一步反应。LCMS:RT＝0.73min,[M-H] -＝205.04。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump to obtain 240 mg of yellow oil N-(methylsulfonyl)piperidine-4-carboxamide trifluoroacetate. It is used directly in the next step without purification. reaction. LCMS: RT=0.73 min, [MH] - =205.04.
步骤C：合成(S)-5-(3-(4-(4-氨基甲酰基哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step C: Synthesis of (S)-5-(3-(4-(4-carbamoylpiperidine-1-carboxamido)phenyl)-2-(2-((5-chloro-2-(1H -Tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000216
Figure PCTCN2020133493-appb-000216
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(120.0毫克，0.16毫摩尔)和N-(甲基磺酰基)哌啶-4-羧酰胺三氟乙酸盐(153.7毫克，0.48毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(124.1毫克，0.96毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (120.0 mg, 0.16 mmol) and N-(methylsulfonyl) Piperidine-4-carboxamide trifluoroacetate (153.7 mg, 0.48 mmol) in tetrahydrofuran (3.0 ml) was added dropwise with N,N-diisopropylethylamine (124.1 mg, 0.96 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＝1/1)。得 到68毫克白色固体(S)-5-(3-(4-(4-氨基甲酰基哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：53.2％)。LCMS:RT＝4.16min,[M+H] +＝798.15。 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 68 mg of white solid (S)-5-(3-(4-(4-carbamoylpiperidine-1-carboxamido)phenyl)-2-(2-((5-chloro-2-( 1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 53.2%). LCMS: RT = 4.16 min, [M+H] + =798.15.
步骤D：合成(S)-5-(3-(4-(4-氨基甲酰基哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸Step D: Synthesis of (S)-5-(3-(4-(4-carbamoylpiperidine-1-carboxamido)phenyl)-2-(2-((5-chloro-2-(1H -Tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000217
Figure PCTCN2020133493-appb-000217
室温下，将(S)-5-(3-(4-(4-氨基甲酰基哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(68.0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-(4-carbamoylpiperidine-1-carboxamido)phenyl)-2-(2-((5-chloro-2-(1H -Tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (68.0 mg, 0.09 mmol) was added to dichloromethane (2.0 mL), add trifluoroacetic acid (0.5 mL) dropwise, and 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42毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((甲基磺酰基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：62.9％)。LCMS:RT＝3.26min,[M+H] +＝742.48。 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 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(4-((methylsulfonyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2- Carboxylic acid (yield: 62.9%). LCMS: RT = 3.26 min, [M+H] + =742.48.
实施例34Example 34
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((甲基磺酰基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-((methylsulfonyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000218
Figure PCTCN2020133493-appb-000218
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-((甲基磺酰基)氨基甲酰基)哌啶-1-甲酸叔丁酯Step A: Synthesis of tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000219
Figure PCTCN2020133493-appb-000219
室温下，将1-(叔丁氧羰基)哌啶-4-羧酸(2.0克，8.7毫摩尔)、甲磺酰胺(752.3毫克，7.9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(4.5克，11.9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(3.1克，23.7毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (2.0 g, 8.7 mmol), methanesulfonamide (752.3 mg, 7.9 mmol) and 2-(7-oxybenzotriazide) Azole)-N,N,N',N'-tetramethylurea hexafluorophosphate (4.5g, 11.9mmol) was added to N,N-dimethylformamide (20.0ml), and N,N was added dropwise -Diisopropylethylamine (3.1 g, 23.7 mmol), after dripping , react under N 2 protection at room temperature for 5 hours.
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(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：二氯甲烷/甲醇＝10/1)。得到1.5克黄色固体4-((甲基磺酰基)氨基甲酰基)哌啶-1-甲酸叔丁酯(收率：62.0％)。LCMS:RT＝3.36min,[M-H] -＝305.08。 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 (50 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). 1.5 g of tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate was obtained as a yellow solid (yield: 62.0%). LCMS: RT = 3.36 min, [MH] - =305.08.
步骤B：合成N-(甲基磺酰基)哌啶-4-羧酰胺三氟乙酸盐Step B: Synthesis of N-(methylsulfonyl)piperidine-4-carboxamide trifluoroacetate
Figure PCTCN2020133493-appb-000220
Figure PCTCN2020133493-appb-000220
室温下，将4-((甲基磺酰基)氨基甲酰基)哌啶-1-甲酸叔丁酯(1.5克，5.0毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。At room temperature, tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate (1.5 g, 5.0 mmol) was added to dichloromethane (2.0 mL), and trifluoroacetic acid ( 1.0 ml), 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得到500毫克黄色油状物N-(甲基磺酰基)哌啶-4-羧酰胺三氟乙酸盐，无需纯化，直接用于下一步反应。LCMS:RT＝0.73min,[M-H] -＝205.04。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump to obtain 500 mg of yellow oil N-(methylsulfonyl)piperidine-4-carboxamide trifluoroacetate. It is used directly in the next step without purification. reaction. LCMS: RT=0.73 min, [MH] - =205.04.
步骤C：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((甲基磺酰基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step C: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-((Methylsulfonyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000221
Figure PCTCN2020133493-appb-000221
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(120.0毫克，0.16毫摩尔)和N-(甲基磺酰基)哌啶-4-羧酰胺三氟乙酸盐(153.7毫克，0.48毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(124.1毫克，0.96毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (120.0 mg, 0.16 mmol) and N-(methylsulfonyl) Piperidine-4-carboxamide trifluoroacetate (153.7 mg, 0.48 mmol) in tetrahydrofuran (3.0 ml) was added dropwise with N,N-diisopropylethylamine (124.1 mg, 0.96 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＝1/1)。得到75毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((甲基磺酰基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：53.5％)。LCMS:RT＝3.68min,[M-H] -＝875.37。 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=1/1). Obtain 75 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-((methylsulfonyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield : 53.5%). LCMS: RT = 3.68 min, [MH] - =875.37.
步骤D：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((甲基磺酰基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step D: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-((methylsulfonyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000222
Figure PCTCN2020133493-appb-000222
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((甲基磺酰基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(75.0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 (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-((methylsulfonyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (75.0 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48毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((甲基磺酰基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：65.6％)。LCMS:RT＝3.38min,[M-H] -＝818.18。 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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(4-((methylsulfonyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2- Carboxylic acid (yield: 65.6%). LCMS: RT = 3.38 min, [MH] - =818.18.
实施例35Example 35
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-羟基氮杂环丁烷-1-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(3-Hydroxyazetidine-1-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000223
Figure PCTCN2020133493-appb-000223
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(3-羟基氮杂环丁烷-1-羰基)哌啶-1-甲酸叔丁酯Step A: Synthesis of tert-butyl 4-(3-hydroxyazetidine-1-carbonyl)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000224
Figure PCTCN2020133493-appb-000224
室温下，将1-(叔丁氧羰基)哌啶-4-羧酸(2.0克，8.7毫摩尔)、甲磺酰胺(861.1毫克，7.9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(4.5克，11.9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(3.1克，23.7毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (2.0 g, 8.7 mmol), methanesulfonamide (861.1 mg, 7.9 mmol) and 2-(7-oxybenzotriazide) Azole)-N,N,N',N'-tetramethylurea hexafluorophosphate (4.5g, 11.9mmol) was added to N,N-dimethylformamide (20.0ml), and N,N was added dropwise -Diisopropylethylamine (3.1 g, 23.7 mmol), after dripping , react under N 2 protection at room temperature for 5 hours.
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(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：二氯甲烷/甲醇＝10/1)。得到1.4克黄色固体4-(3-羟基氮杂环丁烷-1-羰基)哌啶-1-甲酸叔丁酯(收率：62.4％)。LCMS:RT＝1.53min,[M-H] -＝283.12。 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 (50 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). 1.4 g of tert-butyl 4-(3-hydroxyazetidine-1-carbonyl)piperidine-1-carboxylate was obtained as a yellow solid (yield: 62.4%). LCMS: RT=1.53 min, [MH] - =283.12.
步骤B：合成(3-羟基氮杂环丁烷-1-基)(哌啶-4-基)甲酮三氟乙酸盐Step B: Synthesis of (3-hydroxyazetidin-1-yl)(piperidin-4-yl)methanone trifluoroacetate
Figure PCTCN2020133493-appb-000225
Figure PCTCN2020133493-appb-000225
室温下，将4-((甲基磺酰基)氨基甲酰基)哌啶-1-甲酸叔丁酯(1.4克，4.9毫摩尔)加入二氯甲烷(2.0毫升)中，滴加三氟乙酸(1.0毫升)，室温反应3小时。At room temperature, tert-butyl 4-((methylsulfonyl)carbamoyl)piperidine-1-carboxylate (1.4 g, 4.9 mmol) was added to dichloromethane (2.0 mL), and trifluoroacetic acid ( 1.0 ml), 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得到475毫克黄色油状物(3-羟基氮杂环丁烷-1-基)(哌啶-4-基)甲酮三氟乙酸盐，无需纯化，直接用于下一步反应。LCMS:RT＝0.67min,[M-H] -＝183.11。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is drained by an oil pump to obtain 475 mg of yellow oil (3-hydroxyazetidine-1-yl) (piperidin-4-yl) ketone trifluoroacetate. , Without purification, directly used in the next reaction. LCMS: RT=0.67 min, [MH] - =183.11.
步骤C：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-羟基氮杂环丁烷-1-羰基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step C: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(3-Hydroxyazetidine-1-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000226
Figure PCTCN2020133493-appb-000226
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(120.0毫克，0.16毫摩尔)和(3-羟基氮杂环丁烷-1-基)(哌啶-4-基)甲酮三氟乙酸盐(135.4毫克，0.48毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(124.1毫克，0.96毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (120.0 mg, 0.16 mmol) and (3-hydroxyazetidine Alk-1-yl)(piperidin-4-yl)methanone trifluoroacetate (135.4 mg, 0.48 mmol) in tetrahydrofuran (3.0 ml) was added dropwise N,N-diisopropylethylamine (124.1 Mg, 0.96 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：二氯甲烷/甲醇＝4/1)。得到57毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-羟基氮杂环丁烷-1-羰基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：41.7％)。LCMS:RT＝3.67min,[M-H] -＝854.31。 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=4/1). Obtain 57 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(3-hydroxyazetidine-1-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (Yield: 41.7%). LCMS: RT = 3.67 min, [MH] - =854.31.
步骤D：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-羟基氮杂环丁烷-1-羰基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step D: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(3-Hydroxyazetidine-1-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000227
Figure PCTCN2020133493-appb-000227
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-羟基氮杂环丁烷-1-羰基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(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, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(3-Hydroxyazetidine-1-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid 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.
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(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(3-羟基氮杂环丁烷-1-羰基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：57.3％)。LCMS:RT＝3.19min,[M+H] +＝798.20。 1H NMR(400MHz,DMSO)δ12.85(s,1H),11.73(s,1H),10.74(s,1H),10.05(s,1H),9.84(s,1H),8.78(d,J＝8.4Hz,1H),8.42(s,1H),7.99(d,J＝2.4Hz,1H),7.95(d,J＝1.7Hz,1H),7.77(d,J＝8.6Hz,1H),7.62(dd,J＝8.6,2.4Hz,1H),7.48–7.27(m,4H),7.12(d,J＝8.7Hz,2H),7.06(d,J＝2.7Hz,1H),4.72–4.63(m,1H),4.53–4.41(m,2H),4.41–4.32(m,2H),4.14–3.98(m,4H),3.90(dd,J＝9.2,4.3Hz,1H),3.56(dd,J＝10.2,4.1Hz,1H),3.08(d,J＝5.5Hz,2H),1.61(d,J＝13.6Hz,2H),1.50–1.35(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 32 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(4-(3-hydroxyazetidine-1-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole -2-carboxylic acid (yield: 57.3%). LCMS: RT = 3.19 min, [M+H] + = 798.20. 1 H NMR (400MHz, DMSO) δ 12.85 (s, 1H), 11.73 (s, 1H), 10.74 (s, 1H), 10.05 (s, 1H), 9.84 (s, 1H), 8.78 (d, J = 8.4 Hz, 1H), 8.42 (s, 1H), 7.99 (d, J = 2.4 Hz, 1H), 7.95 (d, J = 1.7 Hz, 1H), 7.77 (d, J = 8.6 Hz, 1H), 7.62 (dd, J = 8.6, 2.4 Hz, 1H), 7.48–7.27 (m, 4H ), 7.12(d,J=8.7Hz,2H), 7.06(d,J=2.7Hz,1H), 4.72–4.63(m,1H),4.53–4.41(m,2H), 4.41–4.32(m, 2H), 4.14–3.98 (m, 4H), 3.90 (dd, J = 9.2, 4.3 Hz, 1H), 3.56 (dd, J = 10.2, 4.1 Hz, 1H), 3.08 (d, J = 5.5 Hz, 2H ), 1.61 (d, J = 13.6 Hz, 2H), 1.50-1.35 (m, 2H).
实施例36Example 36
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(3-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 3-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000228
Figure PCTCN2020133493-appb-000228
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-3-(3-溴苯基)-2-((叔丁氧羰基)氨基)丙酸Step A: Synthesis of (S)-3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid
Figure PCTCN2020133493-appb-000229
Figure PCTCN2020133493-appb-000229
将(S)-2-氨基-3-(3-溴苯基)丙酸(2.0克，8.20毫摩尔)，二碳酸二叔丁酯(1.97克，9.02毫摩尔)和氢氧化钠(656毫克，16.40毫摩尔)溶解于四氢呋喃(20毫升)和水(10毫升)的混合溶剂中，室温搅拌过夜。Combine (S)-2-amino-3-(3-bromophenyl) propionic acid (2.0 g, 8.20 mmol), di-tert-butyl dicarbonate (1.97 g, 9.02 mmol) and sodium hydroxide (656 mg , 16.40 mmol) was dissolved in a mixed solvent of tetrahydrofuran (20 mL) and water (10 mL), and stirred at room temperature overnight.
用1M盐酸将反应液淬灭，调节pH至3左右，用乙酸乙酯(200毫升)将反应液稀释，分别用水(30毫升×2次)和饱和食盐水(30毫升)洗，有机相用无水硫酸钠干燥，过滤浓缩，所得粗品(S)-3-(3-溴苯基)-2-((叔丁氧羰基)氨基)丙酸直接用于下一步反应。MS(ESI)M/Z：342.04[M-H] -。 The reaction solution was quenched with 1M hydrochloric acid, adjusted to pH 3, diluted with ethyl acetate (200 ml), washed with water (30 ml × 2 times) and saturated brine (30 ml), and used for the organic phase After drying with anhydrous sodium sulfate, filtering and concentrating, the crude product (S)-3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid was directly used in the next reaction. MS (ESI) M/Z: 342.04 [MH] - .
步骤B：合成(S)-4-(3-(3-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基苯甲酸叔丁酯Step B: Synthesis of tert-butyl (S)-4-(3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamidobenzoate
Figure PCTCN2020133493-appb-000230
Figure PCTCN2020133493-appb-000230
将S)-3-(3-溴苯基)-2-((叔丁氧羰基)氨基)丙酸粗品和对氨基苯甲酸叔丁酯(1.90克，9.84毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(4.68克，12.30毫摩尔)和N,N-二异丙基乙胺(4.07毫升，24.60毫摩尔)溶解于N,N-二甲基甲酰胺(10毫升)，室温搅拌过夜。S)-3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid crude product and tert-butyl p-aminobenzoate (1.90 g, 9.84 mmol), 2-(7- Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (4.68g, 12.30mmol) and N,N-diisopropylethylamine (4.07ml, 24.60 mmol) was dissolved in N,N-dimethylformamide (10 mL) and stirred at room temperature overnight.
将反应液用乙酸乙酯(200毫升)稀释，分别用水(30毫升×2次)和饱和食盐水(30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＝5/1)。得到3.65克白色固体(S)-4-(3-(3-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基苯甲酸叔丁酯(收率：两步85.7％)。MS(ESI)M/Z：519.14[M+H] +。 The reaction solution was diluted with ethyl acetate (200 ml), respectively, with water (30 ml × 2 times) and saturated brine (30 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 = 5/1). Obtain 3.65 g of white solid (S)-4-(3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamidobenzoate tert-butyl ester (yield: 85.7% in two steps) .MS(ESI)M/Z: 519.14[M+H] + .
步骤C：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(3-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamide Yl) tert-butyl benzoate
Figure PCTCN2020133493-appb-000231
Figure PCTCN2020133493-appb-000231
将(S)-4-(3-(3-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基苯甲酸叔丁酯(604毫克，1.17毫摩尔)，4-甲基哌嗪-2-酮(266毫克，2.33毫摩尔)，碘化亚铜(222毫克，1.00毫摩尔)，碳酸铯(760毫克，2.33毫摩尔)和N,N-二甲基乙二胺(206毫克，233毫摩尔)溶解于1,4-二氧六环(6毫升)，氮气保护，120摄氏度条件下搅拌过夜。(S)-4-(3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido benzoate (604 mg, 1.17 mmol), 4-methyl Piperazine-2-one (266 mg, 2.33 mmol), cuprous iodide (222 mg, 1.00 mmol), cesium carbonate (760 mg, 2.33 mmol) and N,N-dimethylethylenediamine ( 206 mg, 233 mmol) was dissolved in 1,4-dioxane (6 ml), protected by nitrogen, and stirred overnight at 120 degrees Celsius.
将反应液用乙酸乙酯(200毫升)稀释，分别用水(30毫升×2次)和饱和食盐水(30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(二氯甲烷：甲醇＝10/1)。得到479毫克白色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(3-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：74.4％)。MS(ESI)M/Z：553.30[M+H] +。 The reaction solution was diluted with ethyl acetate (200 ml), respectively, with water (30 ml × 2 times) and saturated brine (30 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 (dichloromethane: methanol = 10/1). Obtain 479 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propane Amido) tert-butyl benzoate (yield: 74.4%). MS (ESI) M/Z: 553.30 [M+H] + .
步骤D：合成(S)-4-(2-氨基-3-(3-(4-甲基-2-氧代哌嗪-1-基)苯基)丙酰胺基叔丁基)苯甲酸叔丁酯Step D: Synthesis of tert-(S)-4-(2-amino-3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido tert-butyl)benzoic acid Butyl
Figure PCTCN2020133493-appb-000232
Figure PCTCN2020133493-appb-000232
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(3-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(205毫克，0.37毫摩尔)溶解于乙酸乙酯(2.24毫升)，室温条件下缓慢滴加2.0M盐酸乙酸乙酯溶液(557微升)，室温条件下搅拌反应2小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzene Tert-butyl formate (205 mg, 0.37 mmol) was dissolved in ethyl acetate (2.24 ml), 2.0M hydrochloric acid ethyl acetate solution (557 μl) was slowly added dropwise at room temperature, and the reaction was stirred at room temperature for 2 hours.
反应液用胺甲醇溶液淬灭至碱性，所得混合溶液将溶剂旋干后用制备硅胶板分离(二氯甲烷：甲醇＝10/1)。得到100毫克(S)-4-(2-氨基-3-(3-(4-甲基-2-氧代哌嗪-1-基)苯基)丙酰胺基叔丁基)苯甲酸叔丁酯(收率：59.6％)。MS(ESI)M/Z：453.30[M+H] +。 The reaction solution was quenched to alkaline with amine methanol solution, and the resulting mixed solution was spin-dried to dry the solvent and separated with a preparative silica gel plate (dichloromethane: methanol = 10/1). Obtain 100 mg of (S)-4-(2-amino-3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido tert-butyl)benzoic acid tert-butyl Ester (yield: 59.6%). MS (ESI) M/Z: 453.30 [M+H] + .
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(3-(4-甲基-2-氧哌嗪-1-基)苯基)丙 酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000233
Figure PCTCN2020133493-appb-000233
将(S)-4-(2-氨基-3-(3-(4-甲基-2-氧代哌嗪-1-基)苯基)丙酰胺基叔丁基)苯甲酸叔丁酯(100毫克，0.22毫摩尔)，2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(54毫克，0.20毫摩尔)，1-丙基磷酸酐(210毫克，0.66毫摩尔)和N,N-二异丙基乙胺(110微升，0.66毫摩尔)溶解于N,N-二甲基甲酰胺(1毫升)，室温搅拌过夜。Add (S)-4-(2-amino-3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido tert-butyl) benzoic acid tert-butyl ester ( 100 mg, 0.22 mmol), 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (54 mg, 0.20 mmol), 1-propane Phosphoric anhydride (210 mg, 0.66 mmol) and N,N-diisopropylethylamine (110 μl, 0.66 mmol) were dissolved in N,N-dimethylformamide (1 mL) and stirred at room temperature 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(二氯甲烷：甲醇＝10/1)。得到50毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(3-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：35.2％)。MS(ESI)M/Z：702.30[M+H] +。 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 (dichloromethane: methanol = 10/1). Obtain 50 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) Tert-Butyl-3-(3-(4-methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate (yield: 35.2%). MS (ESI) M/Z: 702.30 [M+H] + .
步骤F：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(3-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(3-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000234
Figure PCTCN2020133493-appb-000234
将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(3-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(50毫克，0.071毫摩尔)和三乙基硅烷(21毫克，0.178毫摩尔)溶解于二氯甲烷(1.0毫升)，室温条件下缓慢滴加三氟乙酸(0.2毫升)，室温条件反应3小时。(S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 3-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (50 mg, 0.071 mmol) and triethylsilane (21 mg, 0.178 mmol) ) Was dissolved in dichloromethane (1.0 ml), trifluoroacetic acid (0.2 ml) was slowly added dropwise at room temperature, and the reaction was carried out at room temperature for 3 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备硅胶板分离(二氯甲烷：甲醇＝10/1)。得到20毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(3-(4-甲基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸(收率：21.7％)。MS(ESI)M/Z：646.20[M+H] +。 1H NMR(400MHz,DMSO)δ10.77–10.58(m,2H),9.85(s,1H),9.01(s,1H),7.99(s,1H),7.90(d,J＝8.7Hz,2H),7.77(d,J＝8.6Hz,1H),7.72(s,2H),7.62(s,1H),7.31(t,J＝7.6Hz,1H),7.24(d,J＝14.1Hz,2H),7.16(d,J＝7.9Hz,1H),5.77(s,1H),4.74(s,2H),3.53(dd,J＝14.0,5.6Hz,3H),3.19-3.14(m,2H),3.08(s,2H),2.73–2.62(m,2H),2.27(s,3H)。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated with a preparative silica gel plate (dichloromethane: methanol = 10/1). Obtain 20 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(3-(4-Methyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid (yield: 21.7%). MS (ESI) M/Z: 646.20 [M +H] + . 1 H NMR(400MHz,DMSO)δ10.77-10.58(m,2H),9.85(s,1H),9.01(s,1H),7.99(s,1H),7.90(d,J =8.7Hz,2H),7.77(d,J=8.6Hz,1H),7.72(s,2H),7.62(s,1H),7.31(t,J=7.6Hz,1H),7.24(d,J = 14.1Hz, 2H), 7.16 (d, J = 7.9 Hz, 1H), 5.77 (s, 1H), 4.74 (s, 2H), 3.53 (dd, J = 14.0, 5.6 Hz, 3H), 3.19-3.14 (m, 2H), 3.08 (s, 2H), 2.73–2.62 (m, 2H), 2.27 (s, 3H).
实施例37Example 37
合成(S)-4-(2-(2-((5-氯-2-(2,2,2-三氟乙氧基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000235
Figure PCTCN2020133493-appb-000235
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-氯-2-硝基-1-(2,2,2-三氟乙氧基)苯Step A: Synthesis of 4-chloro-2-nitro-1-(2,2,2-trifluoroethoxy)benzene
Figure PCTCN2020133493-appb-000236
Figure PCTCN2020133493-appb-000236
将4-氯-1-氟-2-硝基苯(2.10克，12.0毫摩尔)，2,2,2-三氟乙醇(1.44克，14.4毫摩尔)和碳酸铯(7.82克，24.0毫摩尔)溶解于四氢呋喃(20毫升)，室温搅拌过夜。Combine 4-chloro-1-fluoro-2-nitrobenzene (2.10 g, 12.0 mmol), 2,2,2-trifluoroethanol (1.44 g, 14.4 mmol) and cesium carbonate (7.82 g, 24.0 mmol) ) Was dissolved in tetrahydrofuran (20 mL) and stirred at room temperature overnight.
将反应液用水(10毫升)淬灭，将四氢呋喃用旋蒸拉干，剩余水溶液用乙酸乙酯(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，所得粗品直接用于下一步反应。The reaction solution was quenched with water (10 mL), the tetrahydrofuran was evaporated to dryness, the remaining aqueous solution was diluted with ethyl acetate (100 mL), and the mixture was washed with water (20 mL×2 times) and saturated brine (20 mL). The phase was dried with anhydrous sodium sulfate, filtered and concentrated, and the obtained crude product was directly used in the next reaction.
步骤B：合成4-氯-2-氨基-1-(2,2,2-三氟乙氧基)苯Step B: Synthesis of 4-chloro-2-amino-1-(2,2,2-trifluoroethoxy)benzene
Figure PCTCN2020133493-appb-000237
Figure PCTCN2020133493-appb-000237
将4-氯-2-硝基-1-(2,2,2-三氟乙氧基)苯粗品，二氯化锡二水合物(10.8克，48.0毫摩尔)溶解于乙酸乙酯(20毫升)，室温条件下搅拌反应过夜。The crude 4-chloro-2-nitro-1-(2,2,2-trifluoroethoxy)benzene, tin dichloride dihydrate (10.8 g, 48.0 mmol) was dissolved in ethyl acetate (20 Ml), the reaction was stirred overnight at room temperature.
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，所得粗品直接用于下一步反应。The reaction solution was 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, and the crude product obtained was used directly in the next step reaction.
步骤C：合成2-((5-氯-2-(2,2,2-三氟乙氧基)苯基)氨基)-2-氧代乙酸Step C: Synthesis of 2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000238
Figure PCTCN2020133493-appb-000238
将4-氯-2-氨基-1-(2,2,2-三氟乙氧基)苯粗品，草酰氯单甲酯(883微升，9.6毫摩尔)，吡啶(968微升，12.0毫摩尔)溶解于二氯甲烷(25毫升)，室温搅拌反应5小时。The crude 4-chloro-2-amino-1-(2,2,2-trifluoroethoxy)benzene, monomethyl oxalyl chloride (883 μl, 9.6 mmol), pyridine (968 μl, 12.0 mmol) Mol) was dissolved in dichloromethane (25 ml), stirred at room temperature for 5 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，所得粗品和氢氧化锂(1.0克，24.0毫摩尔)溶解于四氢呋喃(30毫升)/水(10毫升)混合溶液，室温反应5小时。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 the crude product and lithium hydroxide ( 1.0 g, 24.0 mmol) was dissolved in a mixed solution of tetrahydrofuran (30 mL)/water (10 mL), and reacted at room temperature for 5 hours.
将反应液中四氢呋喃拉干，用乙酸乙酯洗(5毫升×2次)。用1M稀盐酸将pH调节至2左右，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＝15/1)。得到150毫克白色固体2-((5-氯-2-(2,2,2-三氟乙氧基)苯基)氨基)-2-氧代乙酸(收率：三步4.2％)。MS(ESI)M/Z：296.00[M-H] -。 The tetrahydrofuran in the reaction solution was pulled dry and washed with ethyl acetate (5 ml×2 times). Adjust the pH to about 2 with 1M dilute hydrochloric acid, dilute with ethyl acetate (100 ml), water (20 ml × 2 times) and saturated brine (20 ml) respectively, dry the organic phase with anhydrous sodium sulfate, filter and concentrate The obtained crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15/1). 150 mg of white solid 2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetic acid was obtained (yield: 4.2% in three steps). MS (ESI) M/Z: 296.00 [MH] - .
步骤D：合成(S)-4-(2-(2-((5-氯-2-(2,2,2-三氟乙氧基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetamido )-3-Phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000239
Figure PCTCN2020133493-appb-000239
将2-((5-氯-2-(2,2,2-三氟乙氧基)苯基)氨基)-2-氧代乙酸(71毫克，0.24毫摩尔)，(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(82毫克，0.24毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(182毫克，0.48毫摩尔)和N,N-二异丙基乙胺(167微升，0.96毫摩尔)溶解于N,N-二甲基甲酰胺(1.5毫升)，室温搅拌过夜。The 2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetic acid (71 mg, 0.24 mmol), (S)-4- (2-Amino-3-phenylpropionamido) tert-butyl benzoate (82 mg, 0.24 mmol), 2-(7-azobenzotriazole)-N,N,N',N' -Tetramethylurea hexafluorophosphate (182 mg, 0.48 mmol) and N,N-diisopropylethylamine (167 μl, 0.96 mmol) were dissolved in N,N-dimethylformamide (1.5 Ml), stirring at room temperature 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＝10/1)。得到80毫克白色固体(S)-4-(2-(2-((5-氯-2-(2,2,2-三氟乙氧基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：54.1％)。MS(ESI)M/Z：620.20[M+H] +。 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 = 10/1). Obtain 80 mg of white solid (S)-4-(2-(2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetamide (Yl)-3-phenylpropionamido) tert-butyl benzoate (yield: 54.1%). MS (ESI) M/Z: 620.20 [M+H] + .
步骤E：合成(S)-4-(2-(2-((5-氯-2-(2,2,2-三氟乙氧基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetamido )-3-Phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000240
Figure PCTCN2020133493-appb-000240
将(S)-4-(2-(2-((5-氯-2-(2,2,2-三氟乙氧基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(80毫克，0.13毫摩尔)和三氟乙酸(0.5毫升)溶解于二氯甲烷(2.5毫升)，室温搅拌2小时。(S)-4-(2-(2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido) tert-butyl benzoate (80 mg, 0.13 mmol) and trifluoroacetic acid (0.5 mL) were dissolved in dichloromethane (2.5 mL) and stirred at room temperature for 2 hours.
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30毫克白色固体(S)-4-(2-(2-((5-氯-2-(2,2,2-三氟乙氧基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：41.2％)。LCMS：RT＝4.0min，[M-H] -＝562.10。 1H NMR(400MHz,DMSO)δ12.77(s,1H),10.47(s,1H),9.77(s,1H),9.27(d,J＝8.3Hz,1H),8.19(d,J＝2.0Hz,1H),7.98–7.86(m,2H),7.72(d,J＝8.8Hz,2H),7.40–7.25(m,6H),7.20(t,J＝7.1Hz,1H),4.91(q,J＝9.0Hz,2H),4.78(td,J＝8.8,6.0Hz,1H),3.25–3.18(m,2H)。 Concentrate the reaction solution with a rotary evaporator, and dissolve it with methanol after being further drained by an oil pump, and then add n-hexane to the solution system dropwise, a large amount of solids precipitate out, continue stirring at room temperature for 1 hour, and filter to obtain 30 mg of white solid (S) -4-(2-(2-((5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)amino)-2-oxoacetamido)-3-phenylpropane Amido)benzoic acid (yield: 41.2%). LCMS: RT = 4.0 min, [MH] - =562.10. 1 H NMR (400MHz, DMSO) δ 12.77 (s, 1H), 10.47 (s, 1H), 9.77 (s, 1H), 9.27 (d, J = 8.3 Hz, 1H), 8.19 (d, J = 2.0 Hz, 1H), 7.98–7.86 (m, 2H), 7.72 (d, J = 8.8 Hz, 2H), 7.40–7.25 (m, 6H), 7.20 (t, J = 7.1 Hz, 1H), 4.91 (q , J = 9.0 Hz, 2H), 4.78 (td, J = 8.8, 6.0 Hz, 1H), 3.25-3.18 (m, 2H).
实施例38Example 38
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000241
Figure PCTCN2020133493-appb-000241
步骤A：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000242
Figure PCTCN2020133493-appb-000242
将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(106毫克，0.139毫摩尔)，4-氰基-哌啶盐酸盐(41毫克，0.277毫摩尔)和三乙胺(58微升，0.416毫摩尔)溶解于四氢呋喃(2毫升)，60摄氏度条件下搅拌过夜。Add (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((Phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (106 mg, 0.139 mmol), 4-cyano-piperidine hydrochloride (41 Mg, 0.277 mmol) and triethylamine (58 μl, 0.416 mmol) were dissolved in tetrahydrofuran (2 mL) and stirred overnight at 60 degrees Celsius.
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60毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：55.4％)。LCMS：RT＝3.83min，[M-H] -＝778.29。 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 60 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 55.4%). LCMS: RT = 3.83 min, [MH] - =778.29.
步骤B：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧酰胺基)苯基) 丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000243
Figure PCTCN2020133493-appb-000243
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(60毫克，0.077毫摩尔)溶解于二氯甲烷(5毫升)，室温条件下缓慢滴加入三氟乙酸(1毫升)，继续搅拌反应3小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (60 mg, 0.077 mmol) was dissolved in dichloromethane ( 5 ml), trifluoroacetic acid (1 ml) was slowly added dropwise at room temperature, and the reaction was continued with stirring for 3 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备硅胶板分离纯化(石油醚：乙酸乙酯＝1/5)。得到20毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：35.9％)。LCMS：RT＝3.35min，[M-H] -＝722.26。 1H NMR(400MHz,DMSO)δ12.92(s,1H),11.71(s,1H),10.72(s,1H),9.98(d,J＝39.4Hz,1H),9.83(s,1H),8.75(d,J＝8.4Hz,1H),8.47(s,1H),8.03–7.90(m,1H),7.77(d,J＝8.6Hz,1H),7.61(dd,J＝8.6,2.3Hz,1H),7.42–7.26(m,3H),7.12(d,J＝8.5Hz,1H),7.05(d,J＝1.5Hz,1H),4.67(dd,J＝14.2,7.9Hz,1H),3.68(dd,J＝9.8,6.2Hz,2H),3.25(dd,J＝11.6,7.8Hz,2H),3.07(d,J＝4.7Hz,2H),2.57–2.53(m,1H),2.00(dd,J＝14.7,7.0Hz,1H),1.86(d,J＝6.5Hz,1H),1.66(dt,J＝13.4,7.1Hz,1H)。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated and purified with a preparative silica gel plate (petroleum ether: ethyl acetate = 1/5). Obtain 20 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 35.9%). LCMS: RT = 3.35 min, [MH] - = 722.26. 1 H NMR (400MHz, DMSO) δ 12.92 (s, 1H), 11.71 (s, 1H), 10.72 (s, 1H), 9.98 (d, J = 39.4 Hz, 1H ),9.83(s,1H),8.75(d,J=8.4Hz,1H),8.47(s,1H),8.03-7.90(m,1H),7.77(d,J=8.6Hz,1H),7.61 (dd,J=8.6,2.3Hz,1H),7.42–7.26(m,3H),7.12(d,J=8.5Hz,1H), 7.05(d,J=1.5Hz,1H), 4.67(dd, J = 14.2, 7.9 Hz, 1H), 3.68 (dd, J = 9.8, 6.2 Hz, 2H), 3.25 (dd, J = 11.6, 7.8 Hz, 2H), 3.07 (d, J = 4.7 Hz, 2H), 2.57-2.53 (m, 1H), 2.00 (dd, J=14.7, 7.0 Hz, 1H), 1.86 (d, J=6.5 Hz, 1H), 1.66 (dt, J=13.4, 7.1 Hz, 1H).
实施例39Example 39
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-吗啉基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-morpholinylpiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000244
Figure PCTCN2020133493-appb-000244
步骤A：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-吗啉基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-morpholinylpiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000245
Figure PCTCN2020133493-appb-000245
将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(60毫克，0.079毫摩尔)，4-哌啶基-吗啉盐酸盐(66毫克，0.316毫摩尔)和N,N-二异丙基乙胺(69微升，0.395毫摩尔)溶解于四氢呋喃(1.5毫升)，65摄氏度条件下搅拌过夜。Add (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((Phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (60 mg, 0.079 mmol), 4-piperidinyl-morpholine hydrochloride ( 66 mg, 0.316 mmol) and N,N-diisopropylethylamine (69 μl, 0.395 mmol) were dissolved in tetrahydrofuran (1.5 mL) and stirred overnight at 65 degrees Celsius.
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(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-吗啉基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯直接用于下一步反应。LCMS：RT＝3.67min，[M+H] +＝840.39。 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 to obtain the crude product (S)-5 -(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-? (Hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester was directly used in the next reaction. LCMS: RT=3.67min, [M+H] + = 840.39.
步骤B：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-吗啉基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-morpholinylpiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000246
Figure PCTCN2020133493-appb-000246
将粗品(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-吗啉基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯溶解于二氯甲烷(5毫升)，室温条件下缓慢滴加入三氟乙酸(1毫升)，继续搅拌反应3小时。The crude product (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3- (4-(4-morpholinylpiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester was dissolved in dichloromethane (5ml) at room temperature Trifluoroacetic acid (1 ml) was slowly added dropwise, and the reaction was continued to be stirred for 3 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备高效液相分离。得到9毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-吗啉基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：两步14.6％)。LCMS：RT＝3.01min，[M+H] +＝784.33。 1H NMR(400MHz,DMSO)δ12.93(s,1H),11.73(s,1H),10.74(s,1H),10.04(s,1H),9.84(s,1H),9.59(s,1H),8.74(d,J＝8.2Hz,1H),8.58(s,1H),7.98(d,J＝2.3Hz,1H),7.93(s,1H),7.77(d,J＝8.6Hz,1H),7.62(dd,J＝8.6,2.3Hz,1H),7.39–7.27(m,3H),7.12(d,J＝8.5Hz,2H),7.05(d,J＝1.7Hz,1H),4.67(dd,J＝14.2,7.6Hz,1H),4.25(d,J＝13.2Hz,2H),4.01(d,J＝12.6Hz,2H),3.64(t,J＝11.9Hz,2H),3.08(d,J＝5.1Hz,3H),2.77(t,J＝12.6Hz,2H),2.07(d,J＝10.2Hz,2H),2.00(dd,J＝14.5,6.8Hz,2H),1.55–1.40(m,2H)。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated by a preparation high performance liquid phase. Obtain 9 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(4-morpholinylpiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 14.6% for two steps). LCMS: RT = 3.01min, [M+H] + = 784.33. 1 H NMR (400MHz, DMSO) δ 12.93 (s, 1H), 11.73 (s, 1H), 10.74 (s, 1H), 10.04 (s, 1H) ), 9.84 (s, 1H), 9.59 (s, 1H), 8.74 (d, J = 8.2 Hz, 1H), 8.58 (s, 1H), 7.98 (d, J = 2.3 Hz, 1H), 7.93 (s ,1H),7.77(d,J=8.6Hz,1H),7.62(dd,J=8.6,2.3Hz,1H),7.39–7.27(m,3H),7.12(d,J=8.5Hz,2H) ,7.05(d,J＝1.7Hz,1H), 4.67(dd,J＝14.2,7.6Hz,1H), 4.25(d,J＝13.2Hz,2H),4.01(d,J＝12.6Hz,2H) , 3.64 (t, J = 11.9 Hz, 2H), 3.08 (d, J = 5.1 Hz, 3H), 2.77 (t, J = 12.6 Hz, 2H), 2.07 (d, J = 10.2 Hz, 2H), 2.00 (dd, J=14.5, 6.8 Hz, 2H), 1.55-1.40 (m, 2H).
实施例40Example 40
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-乙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-Ethyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000247
Figure PCTCN2020133493-appb-000247
步骤A：合成4-乙基-哌啶-2-酮Step A: Synthesis of 4-ethyl-piperidin-2-one
Figure PCTCN2020133493-appb-000248
Figure PCTCN2020133493-appb-000248
将3-氧杂哌嗪-1-羧酸叔丁酯(1.02克，5.1毫摩尔)溶解于二氯甲烷(10毫升)，室温条件下缓慢加入三氟乙酸(4毫升)，继续搅拌反应1.5小时。将反应液浓缩干，所得粗品与碘乙烷(800毫克，5.1毫摩尔)和碳酸钾(1.76克，12.75毫摩尔)溶解于乙腈(10毫升)，氮气保护，50摄氏度条件下搅拌反应4小时。Dissolve tert-butyl 3-oxapiperazine-1-carboxylate (1.02 g, 5.1 mmol) in dichloromethane (10 mL), slowly add trifluoroacetic acid (4 mL) at room temperature, continue to stir and react 1.5 hour. The reaction solution was concentrated to dryness, and the resulting crude product was dissolved in acetonitrile (10 mL) with ethyl iodide (800 mg, 5.1 mmol) and potassium carbonate (1.76 g, 12.75 mmol), under nitrogen protection, and stirred for 4 hours at 50 degrees Celsius. .
将反应液用乙酸乙酯(200毫升)稀释，分别用水(10毫升×2次)和饱和食盐水(10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(二氯甲烷：甲醇＝10/1)。得到390毫克白色固体4-乙基-哌啶-2-酮(收率：两步59.7％)。The reaction solution was diluted with ethyl acetate (200 ml), respectively, with water (10 ml × 2 times) and saturated brine (10 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 (dichloromethane: methanol = 10/1). Obtained 390 mg of white solid 4-ethyl-piperidin-2-one (yield: 59.7% in two steps).
步骤B：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-乙基-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-ethyl-2-oxopiperazin-1-yl)phenyl)propane Amido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000249
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(965毫克，1.86毫摩尔)，4-乙基-哌啶-2-酮(477毫克，3.73毫摩尔)，碘化亚铜(355毫克，1.86毫摩尔)，N,N-二甲基乙二胺(366微升，3.73毫摩尔)和碳酸铯(1.22克，3.73毫摩尔)溶解于甲苯(8毫升)，氮气保护，110摄氏度条件下搅拌过夜。
Figure PCTCN2020133493-appb-000249
(S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (965 mg, 1.86 mmol), 4-ethyl -Piperidin-2-one (477 mg, 3.73 mmol), cuprous iodide (355 mg, 1.86 mmol), N,N-dimethylethylenediamine (366 μl, 3.73 mmol) and Cesium carbonate (1.22 g, 3.73 mmol) was dissolved in toluene (8 mL), protected by nitrogen, and stirred overnight at 110 degrees Celsius.
将反应液用乙酸乙酯(150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(二氯甲烷：甲醇＝10/1)。得到556毫克白色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-乙基-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：52.8％)。LCMS：RT＝3.16min，[M+H] +＝567.28。 The reaction solution was diluted with ethyl acetate (150 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 (dichloromethane: methanol = 10/1). Obtain 556 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-ethyl-2-oxopiperazin-1-yl)phenyl) Propionamido) tert-butyl benzoate (yield: 52.8%). LCMS: RT = 3.16 min, [M+H] + =567.28.
步骤C：合成(S)-4-(2-氨基-3-(4-(4-乙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of tert-butyl (S)-4-(2-amino-3-(4-(4-ethyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000250
Figure PCTCN2020133493-appb-000250
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-乙基-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(556毫克，0.98毫摩尔)溶解于乙酸乙酯(8毫升)，室温条件下滴加入2M盐酸乙酸乙酯溶液(2.0毫升)。室温条件下搅拌反应2小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-ethyl-2-oxopiperazin-1-yl)phenyl)propionamido) Tert-butyl benzoate (556 mg, 0.98 mmol) was dissolved in ethyl acetate (8 mL), and 2M hydrochloric acid ethyl acetate solution (2.0 mL) was added dropwise at room temperature. The reaction was stirred at room temperature for 2 hours.
将反应液过滤得到425毫克白色固体(S)-4-(2-氨基-3-(4-(4-乙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：62.9％)。LCMS：RT＝3.14min，[M+H] +＝467.22。 The reaction solution was filtered to obtain 425 mg of white solid (S)-4-(2-amino-3-(4-(4-ethyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid Tert-butyl ester (yield: 62.9%). LCMS: RT = 3.14 min, [M+H] + =467.22.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-乙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Ethyl-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000251
Figure PCTCN2020133493-appb-000251
将2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(175毫克，0.65毫摩尔)，(S)-4-(2-氨基-3-(4-(4-乙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(270毫克，0.50毫摩尔)，1-丙基磷酸酐(286毫克，0.75毫摩尔)和N,N-二异丙基乙胺(349微升，2.0毫摩尔)溶解于N,N-二甲基甲酰胺(2毫升)，室温搅拌过夜。Add 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (175 mg, 0.65 mmol), (S)-4-(2-amino -3-(4-(4-Ethyl-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (270 mg, 0.50 mmol), 1-propyl phosphoric anhydride ( 286 mg, 0.75 mmol) and N,N-diisopropylethylamine (349 μl, 2.0 mmol) were dissolved in N,N-dimethylformamide (2 mL) and stirred at room temperature 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(二氯甲烷：甲醇＝10/1)。得到90毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-乙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：25.1％)。LCMS：RT＝3.21min，[M+H] +＝716.17。 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 (dichloromethane: methanol = 10/1). Obtain 90 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(4-ethyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate (yield: 25.1%). LCMS: RT=3.21 min, [M+H] + =716.17.
步骤E：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-乙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Ethyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000252
Figure PCTCN2020133493-appb-000252
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-乙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(90毫克，0.13毫摩尔)溶解于二氯甲烷(5毫升)，室温条件下缓慢滴加入三氟乙酸(1毫升)，继续搅拌反应2小时。(S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-Ethyl-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (90 mg, 0.13 mmol) was dissolved in dichloromethane (5 mL) at room temperature Trifluoroacetic acid (1 ml) was slowly added dropwise, and the reaction was continued for 2 hours with stirring.
将反应液用旋转蒸发仪浓缩，粗品用制备高效液相分离。得到30毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-乙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸(收率：36.2％)。LCMS：RT＝2.80min，[M+H] +＝660.19。 1H NMR(400MHz,DMSO)δ12.89–12.67(m,1H),10.74(s,1H),10.50(s,1H),9.83(s,1H),9.06(d,J＝8.2Hz,1H),7.97(d,J＝2.3Hz,1H),7.93(d,J＝8.7Hz,2H),7.78(d,J＝8.6Hz,1H),7.70(d,J＝8.8Hz,2H),7.63(dd,J＝8.7,2.4Hz,1H),7.36(d,J＝8.5Hz,2H),7.29(d,J＝8.5Hz,2H),5.77(s,1H),4.74(dd,J＝14.3,8.2Hz,1H),4.00-3.80(m,2H),3.40-3.15(m,4H),1.30-1.22(m,3H)。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated by a preparation high performance liquid phase. Obtain 30 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-ethyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid (yield: 36.2%). LCMS: RT = 2.80 min, [M+H] + = 660.19. 1 H NMR(400MHz,DMSO)δ12.89-12.67(m,1H), 10.74(s,1H), 10.50(s,1H), 9.83(s,1H), 9.06(d,J=8.2Hz,1H ), 7.97 (d, J = 2.3 Hz, 1H), 7.93 (d, J = 8.7 Hz, 2H), 7.78 (d, J = 8.6 Hz, 1H), 7.70 (d, J = 8.8 Hz, 2H), 7.63(dd,J=8.7,2.4Hz,1H),7.36(d,J=8.5Hz,2H),7.29(d,J=8.5Hz,2H),5.77(s,1H),4.74(dd,J = 14.3, 8.2 Hz, 1H), 4.00-3.80 (m, 2H), 3.40-3.15 (m, 4H), 1.30-1.22 (m, 3H).
实施例41Example 41
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(2-(dimethylamino)ethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000253
Figure PCTCN2020133493-appb-000253
步骤A：合成4-(2-(二甲基氨基)乙氧基)哌啶-1-羧酸叔丁酯Step A: Synthesis of tert-butyl 4-(2-(dimethylamino)ethoxy)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000254
Figure PCTCN2020133493-appb-000254
将4-羟基哌啶-1-羧酸叔丁酯(3.0克，15.0毫摩尔)，和钠氢(2.4克，60.0毫摩尔)溶解于N,N-二甲基甲酰胺(20毫升),0摄氏度搅拌30分钟。将2-溴-N,N-二甲基乙-1-胺氢溴酸盐(4.2克，17.9毫摩尔)加入，室温条件下搅拌反应5小时。Dissolve 4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (3.0 g, 15.0 mmol), and sodium hydrogen (2.4 g, 60.0 mmol) in N,N-dimethylformamide (20 mL), Stir at 0°C for 30 minutes. 2-Bromo-N,N-dimethylethyl-1-amine hydrobromide (4.2 g, 17.9 mmol) was added, and the reaction was stirred at room temperature for 5 hours.
将反应液用水冰淬灭，用乙酸乙酯(300毫升)稀释，分别用水(50毫升×2次)和饱和食盐水(50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(二氯甲烷：甲醇＝10/1)。得到778毫克白色固体4-(2-(二甲基氨基)乙氧基)哌啶-1-羧酸叔丁酯(收率：19.2％)。The reaction solution was quenched with water and ice, diluted with ethyl acetate (300 ml), water (50 ml × 2 times) and saturated brine (50 ml) respectively, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product Purified by silica gel column chromatography (dichloromethane: methanol = 10/1). 778 mg of tert-butyl 4-(2-(dimethylamino)ethoxy)piperidine-1-carboxylate was obtained as a white solid (yield: 19.2%).
步骤B：合成N,N-二甲基-2-(哌啶-4-基氧基)乙-1-胺Step B: Synthesis of N,N-dimethyl-2-(piperidin-4-yloxy)ethan-1-amine
Figure PCTCN2020133493-appb-000255
Figure PCTCN2020133493-appb-000255
将4-(2-(二甲基氨基)乙氧基)哌啶-1-羧酸叔丁酯(778毫克，2.86毫摩尔)和三氟乙酸(4毫升)溶解于二氯甲烷(10毫升)，室温条件下搅拌反应3小时，所得N,N-二甲基-2-(哌啶-4-基氧基)乙-1-胺二三氟乙酸盐粗品直接用于下一步反应。4-(2-(Dimethylamino)ethoxy)tert-butyl piperidine-1-carboxylate (778 mg, 2.86 mmol) and trifluoroacetic acid (4 mL) were dissolved in dichloromethane (10 mL) ), the reaction was stirred at room temperature for 3 hours, and the obtained crude N,N-dimethyl-2-(piperidin-4-yloxy)ethan-1-amine ditrifluoroacetate was directly used in the next reaction.
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-羧酸叔丁酯Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-(dimethylamino)ethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000256
Figure PCTCN2020133493-appb-000256
将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(128毫克，0.168毫摩尔)，N,N-二甲基-2-(哌啶-4-基氧基)乙-1-胺二三氟乙酸盐(201毫克，0.503毫摩尔)和N,N-二异丙基乙胺(234微升，0.395毫摩尔)溶解于四氢呋喃(3.0毫升)，60摄氏度条件下搅拌过夜。Add (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((Phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (128 mg, 0.168 mmol), N,N-dimethyl-2-(piper (Pyridin-4-yloxy)ethyl-1-amine ditrifluoroacetate (201 mg, 0.503 mmol) and N,N-diisopropylethylamine (234 μl, 0.395 mmol) were dissolved in tetrahydrofuran (3.0 ml), stirred overnight at 60 degrees Celsius.
将反应液用乙酸乙酯(100毫升)稀释，分别用水(10毫升×2次)和饱和食盐水(10毫升)，有机相用无水硫酸钠干燥，过滤浓缩，所得粗品(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-羧酸叔丁酯直接用于下一步反应。LCMS：RT＝3.17min，[M-H] -＝840.32。 The reaction solution was diluted with ethyl acetate (100 ml), respectively, with water (10 ml × 2 times) and saturated brine (10 ml), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain crude product (S)-5 -(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-(4-( 2-(Dimethylamino)ethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester was directly used in the next reaction. LCMS: RT = 3.17 min, [MH] - = 840.32.
步骤D：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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 D: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-(dimethylamino)ethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000257
Figure PCTCN2020133493-appb-000257
将粗品(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-羧酸叔丁酯溶解于三氟乙酸(1毫升)和二氯甲烷(5毫升)混合溶剂，室温条件下搅拌反应2小时。The crude product (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3- (4-(4-(2-(Dimethylamino)ethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester is dissolved in three A mixed solvent of fluoroacetic acid (1 mL) and dichloromethane (5 mL) was stirred and reacted at room temperature for 2 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备高效液相分离。得到19毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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：两步14.4％)。LCMS：RT＝2.85min，[M+H] +＝786.22。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated by a preparation high performance liquid phase. Obtain 19 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(4-(2-(Dimethylamino)ethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid (yield : Two steps 14.4%). LCMS: RT = 2.85 min, [M+H] + =786.22.
实施例42Example 42
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000258
Figure PCTCN2020133493-appb-000258
步骤A：合成(S)-2-(((叔丁氧羰基)氨基)叔丁基-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酸酯Step A: Synthesis of (S)-2-(((tert-butoxycarbonyl)amino)tert-butyl-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propane Acid ester
Figure PCTCN2020133493-appb-000259
Figure PCTCN2020133493-appb-000259
将(S)-3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酸叔丁酯(1.06克，2.65毫摩尔)，4-异丙基哌嗪-2-酮(753毫克，5.30毫摩尔)，碘化亚铜(505毫克，2.65毫摩尔)，N,N-二甲基乙二胺(519微升，5.30毫摩尔)和碳酸铯(1.73克，5.30毫摩尔)溶解于甲苯(10毫升)，氮气保护，110摄氏度条件下搅拌过夜。Add (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid tert-butyl ester (1.06 g, 2.65 mmol), 4-isopropylpiperazine-2- Ketone (753 mg, 5.30 mmol), cuprous iodide (505 mg, 2.65 mmol), N,N-dimethylethylenediamine (519 μl, 5.30 mmol) and cesium carbonate (1.73 g, 5.30 MM) was dissolved in toluene (10 mL), protected by nitrogen, and stirred overnight at 110 degrees Celsius.
将反应液用乙酸乙酯(150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(二氯甲烷：甲醇＝10/1)。得到780毫克白色固体(S)-2-(((叔丁氧羰基)氨基)叔丁基-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酸酯(收率：63.8％)。LCMS：RT＝2.98min，[M+Na] +＝484.24。 The reaction solution was diluted with ethyl acetate (150 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 (dichloromethane: methanol = 10/1). Obtain 780 mg of white solid (S)-2-(((tert-butoxycarbonyl)amino)tert-butyl-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl) Propionate (yield: 63.8%). LCMS: RT = 2.98 min, [M+Na] + =484.24.
步骤B：合成(S)-2-((((9H-芴-9-基)甲氧基)羰基)氨基)叔丁基-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸酯Step B: Synthesis of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)tert-butyl-3-(4-(4-isopropyl-2-oxo) Piperazin-1-yl)phenyl)propionate
Figure PCTCN2020133493-appb-000260
Figure PCTCN2020133493-appb-000260
将(S)-2-(((叔丁氧羰基)氨基)叔丁基-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酸酯(680毫克，1.47毫摩尔)溶解于三氟乙酸(1.4毫升)和二氯甲烷(7.0毫升)的混合溶剂，室温搅拌3小时。浓缩所得粗品与氯甲酸-9-芴基甲酯(419毫克，1.62毫摩尔)和碳酸钠(624毫克，5.88毫摩尔)溶解于四氢呋喃(6毫升)和水(6毫升)的混合溶剂，室温搅拌过夜。Add (S)-2-(((tert-butoxycarbonyl)amino)tert-butyl-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionate ( 680 mg, 1.47 mmol) was dissolved in a mixed solvent of trifluoroacetic acid (1.4 mL) and dichloromethane (7.0 mL) and stirred at room temperature for 3 hours. The crude product obtained by concentration was mixed with 9-fluorenyl methyl chloroformate (419 mg, 1.62 mmol) and sodium carbonate (624 mg, 5.88 mmol) were dissolved in a mixed solvent of tetrahydrofuran (6 mL) and water (6 mL), and stirred at room temperature overnight.
将反应液用乙酸乙酯(150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(S)-2-((((9H-芴-9-基)甲氧基)羰基)氨基)叔丁基-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸酯直接用于下一步反应。LCMS：RT＝3.31min，[M+H] +＝584.25。 The reaction solution was diluted with ethyl acetate (150 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 product (S)-2 -((((9H-fluoren-9-yl)methoxy)carbonyl)amino)tert-butyl-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl ) Propionate is directly used in the next reaction. LCMS: RT = 3.31 min, [M+H] + =584.25.
步骤C：合成(S)-2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酸Step C: Synthesis of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazine-1 -(Yl)phenyl)propionic acid
Figure PCTCN2020133493-appb-000261
Figure PCTCN2020133493-appb-000261
将(S)-2-((((9H-芴-9-基)甲氧基)羰基)氨基)叔丁基-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基)丙酸酯粗品溶解于三氟乙酸(3毫升)和二氯甲烷(6毫升)的混合溶剂，室温搅拌反应3小时。浓缩得(S)-2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酸粗品直接用于下一步反应。Add (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)tert-butyl-3-(4-(4-isopropyl-2-oxopiperazine- The crude 1-yl)phenyl)propionate was dissolved in a mixed solvent of trifluoroacetic acid (3ml) and dichloromethane (6ml), and the reaction was stirred at room temperature for 3 hours. Concentrate to obtain (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl The crude) phenyl) propionic acid was directly used in the next reaction.
步骤D：合成(S)-5-(2-氨基-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基叔丁基)-1H-吲哚-2-羧酸酯Step D: Synthesis of (S)-5-(2-amino-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido tert-butyl)-1H- Indole-2-carboxylate
Figure PCTCN2020133493-appb-000262
Figure PCTCN2020133493-appb-000262
将(S)-2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酸粗品(1.47毫摩尔)，5-氨基-1H-吲哚-2-羧酸叔丁酯(488毫克，1.47毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(839毫克，2.21毫摩尔)和N,N-二异丙基乙胺(770微升，4.42毫摩尔)溶解于N,N-二甲基甲酰胺(5毫升)，室温搅拌过夜。(S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl) Crude phenyl)propionic acid (1.47 mmol), tert-butyl 5-amino-1H-indole-2-carboxylate (488 mg, 1.47 mmol), 2-(7-azobenzotriazole) -N,N,N',N'-tetramethylurea hexafluorophosphate (839 mg, 2.21 mmol) and N,N-diisopropylethylamine (770 μl, 4.42 mmol) are dissolved in N , N-dimethylformamide (5 mL), stirred at room temperature 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，所得粗品与4-甲基哌啶(1.5毫升)溶解于二氯甲烷(6.0毫升)，室温搅拌5小时。The reaction solution was 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, the resulting crude product was mixed with 4-methyl Piperidine (1.5 mL) was dissolved in dichloromethane (6.0 mL) and stirred at room temperature for 5 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，所得粗品用硅胶柱层析纯化(二氯甲烷：甲醇＝10/1)。得到610毫克白色固体(S)-5-(2-氨基-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基叔丁基)-1H-吲哚-2-羧酸酯(收率：四步80.0％)。LCMS：RT＝2.83min，[M+H] +＝520.20。 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 (dichloromethane: methanol = 10/1). Obtain 610 mg of white solid (S)-5-(2-amino-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido tert-butyl)-1H -Indole-2-carboxylate (yield: 80.0% in four steps). LCMS: RT = 2.83 min, [M+H] + =520.20.
步骤E：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step E: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000263
Figure PCTCN2020133493-appb-000263
将2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(264毫克，0.99毫摩尔)，(S)-5-(2-氨基-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基叔丁基)-1H-吲哚-2-羧酸酯(610毫克，0.99毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(565毫克，1.49毫摩尔)和N,N-二异丙基乙胺(519微升，2.98毫摩尔)溶解于N,N-二甲基甲酰胺(4毫升)，室温搅拌过夜。Add 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (264 mg, 0.99 mmol), (S)-5-(2-amino -3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido tert-butyl)-1H-indole-2-carboxylate (610 mg, 0.99 mg Mol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (565 mg, 1.49 mmol) and N,N-diiso Propylethylamine (519 μl, 2.98 mmol) was dissolved in N,N-dimethylformamide (4 mL) and stirred at room temperature 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(二氯甲烷：甲醇＝10/1)。得到100毫克(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：13.2)。LCMS：RT＝3.54min，[M+H] +＝769.29。 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 (dichloromethane: methanol = 10/1). Obtain 100 mg of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3- (4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (Yield: 13.2). LCMS: RT = 3.54 min, [M+H] + =769.29.
步骤F：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step F: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000264
Figure PCTCN2020133493-appb-000264
将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(100毫克，0.13毫摩尔)溶解于三氟乙酸(1毫升)和二氯甲烷(5毫升)混合溶剂，室温搅拌1.5小时。Add (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (100 mg, 0.13 mmol) dissolved in trifluoro Acetic acid (1 mL) and dichloromethane (5 mL) were mixed solvents and stirred at room temperature for 1.5 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备高效液相分离。得到20毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：21.6％)。LCMS：RT＝2.82min，[M+H] +＝713.39。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated by a preparation high performance liquid phase. Obtain 20 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 21.6%). LCMS: RT = 2.82 min, [M+H] + = 713.39.
实施例43Example 43
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-羟乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(2-hydroxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000265
Figure PCTCN2020133493-appb-000265
步骤A：合成4-(2-((叔丁基二甲基甲硅烷基)氧基)乙基)哌嗪-2-酮Step A: Synthesis of 4-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperazin-2-one
Figure PCTCN2020133493-appb-000266
Figure PCTCN2020133493-appb-000266
将哌嗪-2-酮三氟乙酸盐(4.0克，18.7毫摩尔)，(2-溴乙氧基)(叔丁基)二甲基硅烷(5.4克，22.4毫摩尔)和碳酸钾(6.5克，46.8毫摩尔)溶解于乙腈(20毫升)，80摄氏度条件下搅拌过夜。Combine piperazine-2-one trifluoroacetate (4.0 g, 18.7 mmol), (2-bromoethoxy) (tert-butyl) dimethylsilane (5.4 g, 22.4 mmol) and potassium carbonate ( 6.5 g, 46.8 mmol) was dissolved in acetonitrile (20 ml) and stirred overnight at 80 degrees Celsius.
将反应液过滤，浓缩后用用硅胶柱层析纯化(二氯甲烷：甲醇＝10/1)。得到1.72克白色固体4-(2-((叔丁基二甲基甲硅烷基)氧基)乙基)哌嗪-2-酮(收率：35.7％)。The reaction solution was filtered, concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 10/1). 1.72 g of white solid 4-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperazin-2-one was obtained (yield: 35.7%).
步骤B：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-(3-((叔丁基二甲基甲硅烷基)氧基)丙基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(3-((tert-butyldimethylsilyl)oxy) (Propyl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000267
Figure PCTCN2020133493-appb-000267
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(982毫克，1.89毫摩尔)，4-(2-((叔丁基二甲基甲硅烷基)氧基)乙基)哌嗪-2-酮(979毫克，3.79毫摩尔)，碘化亚铜(360毫克，1.89毫摩尔)，N,N-二甲基乙二胺(372微升，3.79毫摩尔)和碳酸铯(1.24克，3.79毫摩尔)溶解于甲苯(8毫升)，氮气保护，110摄氏度条件下搅拌过夜。(S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (982 mg, 1.89 mmol), 4-( 2-((tert-butyldimethylsilyl)oxy)ethyl)piperazin-2-one (979 mg, 3.79 mmol), cuprous iodide (360 mg, 1.89 mmol), N, N-Dimethylethylenediamine (372 μl, 3.79 mmol) and cesium carbonate (1.24 g, 3.79 mmol) were dissolved in toluene (8 mL), protected by nitrogen, and stirred overnight at 110 degrees Celsius.
将反应液用乙酸乙酯(150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(二氯甲烷：甲醇＝15/1)。得到1.0克白色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-乙基-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：75.7％)。LCMS：RT＝4.36min，[M+H] +＝697.39。 The reaction solution was diluted with ethyl acetate (150 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 (dichloromethane: methanol = 15/1). Obtain 1.0 g of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-ethyl-2-oxopiperazin-1-yl)phenyl) Propionamido) tert-butyl benzoate (yield: 75.7%). LCMS: RT = 4.36 min, [M+H] + =697.39.
步骤C：合成(S)-4-(2-氨基-3-(4-(4-(2-羟乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-amino-3-(4-(4-(2-hydroxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzene Tert-butyl formate
Figure PCTCN2020133493-appb-000268
Figure PCTCN2020133493-appb-000268
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-乙基-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(1.0克，1.44毫摩尔) 溶解于乙酸乙酯(8.8毫升)，室温条件下滴加入2M盐酸乙酸乙酯溶液，继续搅拌1小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-ethyl-2-oxopiperazin-1-yl)phenyl)propionamido) Tert-butyl benzoate (1.0 g, 1.44 mmol) was dissolved in ethyl acetate (8.8 mL), 2M hydrochloric acid ethyl acetate solution was added dropwise at room temperature, and stirring was continued for 1 hour.
过滤得476毫克白色固体(S)-4-(2-氨基-3-(4-(4-(2-羟乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：55.6％)。Filter to obtain 476 mg of white solid (S)-4-(2-amino-3-(4-(4-(2-hydroxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido group ) Tert-butyl benzoate (yield: 55.6%).
步骤D：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-羟乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(4-(2-hydroxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000269
Figure PCTCN2020133493-appb-000269
将2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(230毫克，0.86毫摩尔)，(S)-4-(2-氨基-3-(4-(4-(2-羟乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(476毫克，0.86毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(490毫克，1.29毫摩尔)和N,N-二异丙基乙胺(600微升，3.44毫摩尔)溶解于N,N-二甲基甲酰胺(5毫升)，室温搅拌过夜。Add 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (230 mg, 0.86 mmol), (S)-4-(2-amino -3-(4-(4-(2-hydroxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (476 mg, 0.86 mmol), 2 -(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (490mg, 1.29mmol) and N,N-diisopropylethylamine (600 μl, 3.44 mmol) was dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature 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(二氯甲烷：甲醇＝10/1)。得到387毫克白色固体(S)-4-(3-(4-(4-(2-(((叔丁基二甲基甲硅烷基)氧基)乙基)-2-氧哌嗪-1-基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)苯甲酸叔丁酯(收率：61.6％)。LCMS：RT＝3.23min，[M+H] +＝732.24。 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 (dichloromethane: methanol = 10/1). Obtain 387 mg of white solid (S)-4-(3-(4-(4-(2-(((tert-butyldimethylsilyl)oxy)ethyl)-2-oxypiperazine-1 -Yl)phenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)tert-benzoic acid Butyl ester (yield: 61.6%). LCMS: RT = 3.23 min, [M+H] + =732.24.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-羟乙基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-hydroxyethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000270
Figure PCTCN2020133493-appb-000270
将(S)-4-(3-(4-(4-(2-(((叔丁基二甲基甲硅烷基)氧基)乙基)-2-氧哌嗪-1-基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)苯甲酸叔丁酯(387毫克，0.53毫摩尔)溶解于三氟乙酸(1毫升)和二氯甲烷(5毫升)混合溶剂，室温搅拌2小时。(S)-4-(3-(4-(4-(2-((((tert-butyldimethylsilyl)oxy)ethyl)-2-oxopiperazin-1-yl)benzene Yl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetylamino)propionamido) tert-butyl benzoate (387 Mg, 0.53 mmol) was dissolved in a mixed solvent of trifluoroacetic acid (1 mL) and dichloromethane (5 mL), and stirred at room temperature for 2 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备高效液相分离。得到40毫克白色固体(S)-4-(3-(4-(4-(2-(((叔丁基二甲基甲硅烷基)氧基)乙基)-2-氧哌嗪-1-基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)苯甲酸(收率：11.2％)。LCMS：RT＝2.81min，[M-H] -＝674.13。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated by a preparation high performance liquid phase. Obtain 40 mg of white solid (S)-4-(3-(4-(4-(2-(((tert-butyldimethylsilyl)oxy)ethyl)-2-oxopiperazine-1 -Yl)phenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)benzoic acid ( Yield: 11.2%). LCMS: RT = 2.81 min, [MH] - =674.13.
实施例44Example 44
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代咪唑烷-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(2-oxoimidazolidine-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000271
Figure PCTCN2020133493-appb-000271
步骤A：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of tertiary (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(2-oxoimidazolin-1-yl)phenyl)propionamido)benzoic acid Butyl
Figure PCTCN2020133493-appb-000272
Figure PCTCN2020133493-appb-000272
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(320毫克，0.62毫摩尔)，咪唑啉-2-酮(107毫克，1.24毫摩尔)，碘化亚铜(119毫克，0.62毫摩尔)，N,N-二甲基乙二胺(122微升，1.24毫摩尔)和碳酸铯(404毫克，1.24毫摩尔)溶解于甲苯(3毫升)，氮气保护，110摄氏度条件下搅拌过夜。Add (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (320 mg, 0.62 mmol), imidazoline- 2-ketone (107 mg, 1.24 mmol), cuprous iodide (119 mg, 0.62 mmol), N,N-dimethylethylenediamine (122 μl, 1.24 mmol) and cesium carbonate (404 mg , 1.24 mmol) was dissolved in toluene (3 ml), protected by nitrogen, and stirred overnight at 110 degrees Celsius.
将反应液用乙酸乙酯(150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(二氯甲烷：甲醇＝15/1)。得到75毫克白色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：23.2％)。LCMS：RT＝4.12min，[M+Na] +＝547.21。 The reaction solution was diluted with ethyl acetate (150 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 (dichloromethane: methanol = 15/1). Obtain 75 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(2-oxoimidazolin-1-yl)phenyl)propionamido)benzoic acid Tert-butyl ester (yield: 23.2%). LCMS: RT = 4.12 min, [M+Na] + =547.21.
步骤B：合成(S)-4-(2-氨基-3-(4-(2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of tert-butyl (S)-4-(2-amino-3-(4-(2-oxoimidazolin-1-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000273
Figure PCTCN2020133493-appb-000273
将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(600毫克，1.15毫摩尔)溶解于乙酸乙酯(6.9毫升)，室温下滴加入2M盐酸乙酸乙酯溶液(1.7毫升)，继续搅拌4小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(2-oxoimidazolin-1-yl)phenyl)propionamido) tert-butyl benzoate ( 600 mg, 1.15 mmol) was dissolved in ethyl acetate (6.9 mL), 2M hydrochloric acid ethyl acetate solution (1.7 mL) was added dropwise at room temperature, and stirring was continued for 4 hours.
将反应液过滤得380毫克白色固体(S)-4-(2-氨基-3-(4-(2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：78.3％)。LCMS：RT＝2.82min，[M+Na] +＝447.22。 The reaction solution was filtered to obtain 380 mg of white solid (S)-4-(2-amino-3-(4-(2-oxoimidazolin-1-yl)phenyl)propionamido) tert-butyl benzoate ( Yield: 78.3%). LCMS: RT = 2.82 min, [M+Na] + =447.22.
步骤C：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代咪唑烷-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(2-oxoimidazolidine-1-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000274
Figure PCTCN2020133493-appb-000274
将2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(221毫克，0.825毫摩尔)，(S)-4-(2-氨基-3-(4-(2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(380毫克，0.825毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(471毫克，1.238毫摩尔)和N,N-二异丙基乙胺(431微升，2.475毫摩尔)溶解于N,N-二甲基甲酰胺(3毫升)，室温搅拌过夜。Add 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (221 mg, 0.825 mmol), (S)-4-(2-amino -3-(4-(2-oxoimidazolin-1-yl)phenyl)propionamido) tert-butyl benzoate (380 mg, 0.825 mmol), 2-(7-azobenzotriazide Azole)-N,N,N',N'-tetramethylurea hexafluorophosphate (471mg, 1.238mmol) and N,N-diisopropylethylamine (431μl, 2.475mmol) dissolved In N,N-dimethylformamide (3 mL), stir overnight at room temperature.
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(二氯甲烷：甲醇＝10/1)。得到132毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代咪唑烷-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：23.7％)。LCMS：RT＝4.03min，[M+H] +＝674.21。 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 (dichloromethane: methanol = 10/1). Obtain 132 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) Tert-Butyl-3-(4-(2-oxoimidazolidine-1-yl)phenyl)propionamido)benzoate (Yield: 23.7%). LCMS: RT=4.03min, [M+H] + = 674.21.
步骤D：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代咪唑烷-1-基)苯基)丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(2-oxoimidazolidine-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000275
Figure PCTCN2020133493-appb-000275
将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代咪唑烷-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(132毫克，0.20毫摩尔)溶解于三氟乙酸(1毫升)和二氯甲烷(5毫升)的混合溶剂，室温搅拌1.5小时。(S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( Tert-Butyl 4-(2-oxoimidazolidine-1-yl)phenyl)propionamido)benzoate (132 mg, 0.20 mmol) was dissolved in trifluoroacetic acid (1 mL) and dichloromethane (5 mL) ), stirring at room temperature for 1.5 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备高效液相分离。得到20毫克(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(2-氧代咪唑烷-1-基)苯基)丙酰胺基)苯甲酸(收率：16.6％)。LCMS：RT＝3.45min，[M-H] -＝616.08。 1H NMR(400MHz,DMSO)δ12.75(s,1H),10.72(s,1H),10.49(s,1H),9.82(d,J＝1.7Hz,1H),8.92(d,J＝8.2Hz,1H),7.97(d,J＝2.3Hz,1H),7.95–7.89(m,2H),7.77(d,J＝8.6Hz,1H),7.74–7.68(m,2H),7.62(dd,J＝8.6,2.4Hz,1H),7.47(d,J＝8.8Hz,1H),7.20(d,J＝8.7Hz,1H),6.90(s,1H),5.76(s,1H),4.69(dd,J＝14.6,8.1Hz,1H),3.82(dd,J＝10.9,7.6Hz,2H),3.39(t,J＝8.2Hz,2H),3.10(d,J＝6.7Hz,2H)。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated by a preparation high performance liquid phase. Obtain 20 mg of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(2-oxoimidazolidine-1-yl)phenyl)propionamido)benzoic acid (yield: 16.6%). LCMS: RT = 3.45 min, [MH] - =616.08. 1 H NMR (400MHz,DMSO)δ12.75(s,1H), 10.72(s,1H), 10.49(s,1H), 9.82(d,J=1.7Hz,1H), 8.92(d,J=8.2Hz,1H ), 7.97(d,J=2.3Hz,1H),7.95-7.89(m,2H),7.77(d,J=8.6Hz,1H),7.74-7.68(m,2H),7.62(dd,J= 8.6, 2.4 Hz, 1H), 7.47 (d, J = 8.8 Hz, 1H), 7.20 (d, J = 8.7 Hz, 1H), 6.90 (s, 1H), 5.76 (s, 1H), 4.69 (dd, J = 14.6, 8.1 Hz, 1H), 3.82 (dd, J = 10.9, 7.6 Hz, 2H), 3.39 (t, J = 8.2 Hz, 2H), 3.10 (d, J = 6.7 Hz, 2H).
实施例45Example 45
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(3-(Cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000276
Figure PCTCN2020133493-appb-000276
步骤A：合成1-(环丙基甲基)咪唑啉-2-酮Step A: Synthesis of 1-(cyclopropylmethyl)imidazolin-2-one
Figure PCTCN2020133493-appb-000277
Figure PCTCN2020133493-appb-000277
将咪唑啉-2-酮(1.02克，11.86毫摩尔)，(溴甲基)环丙烷(1.21毫升，12.45毫摩尔)和钠氢(522毫克，13.05毫摩尔)溶解于N,N-二甲基甲酰胺(10毫升)，氮气保护，35摄氏度条件下搅拌反应过夜。Imidazolin-2-one (1.02 g, 11.86 mmol), (bromomethyl) cyclopropane (1.21 ml, 12.45 mmol) and sodium hydrogen (522 mg, 13.05 mmol) were dissolved in N,N-dimethyl Methyl formamide (10 mL), protected by nitrogen, and stirred overnight at 35 degrees Celsius.
将反应液用乙酸乙酯(200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(二氯甲烷：甲醇＝10/1)。得到400毫克油状物1-(环丙基甲基)咪唑啉-2-酮(收率：24.1％)。The reaction solution was diluted with ethyl acetate (200 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 (dichloromethane: methanol = 10/1). 400 mg of oily 1-(cyclopropylmethyl)imidazolin-2-one was obtained (yield: 24.1%).
步骤B：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-(cyclopropylmethyl)-2-oxoimidazolin-1-yl )Phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000278
Figure PCTCN2020133493-appb-000278
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(847毫克，1.63毫摩尔)，1-(环丙基甲基)咪唑啉-2-酮(400毫克，2.86毫摩尔)，碘化亚铜(311毫克，1.63毫摩尔)，N,N-二甲基乙二胺(320微升，3.27毫摩尔)和碳酸铯(1.07克，3.27毫摩尔)溶解于甲苯(8毫升)，氮气保护，110摄氏度条件下搅拌过夜。(S)-4-(3-(4-Bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (847 mg, 1.63 mmol), 1-( Cyclopropylmethyl)imidazolin-2-one (400 mg, 2.86 mmol), cuprous iodide (311 mg, 1.63 mmol), N,N-dimethylethylenediamine (320 μl, 3.27 (1.07 g, 3.27 mmol) and cesium carbonate (1.07 g, 3.27 mmol) were dissolved in toluene (8 mL), protected by nitrogen, and stirred overnight at 110 degrees Celsius.
将反应液用乙酸乙酯(300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＝2/1)。得到600毫克白色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：63.6％)。LCMS：RT＝2.69min，[M+H] +＝579.46。 The reaction solution was diluted with ethyl acetate (300 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 = 2/1). Obtain 600 mg of white solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-(cyclopropylmethyl)-2-oxoimidazoline-1- (Yl)phenyl)propionamido)tert-butyl benzoate (yield: 63.6%). LCMS: RT = 2.69 min, [M+H] + = 579.46.
步骤C：合成(S)-4-(2-氨基-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-amino-3-(4-(3-(cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenyl)propionamido)benzene Tert-butyl formate
Figure PCTCN2020133493-appb-000279
Figure PCTCN2020133493-appb-000279
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(600毫克，1.04毫摩尔)溶解于乙酸乙酯(10毫升)，室温下滴加入2M盐酸乙酸乙酯溶液(2.5毫升)，继续搅拌反应3小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-(cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenyl ) Propionamido) tert-butyl benzoate (600 mg, 1.04 mmol) was dissolved in ethyl acetate (10 mL), 2M hydrochloric acid ethyl acetate solution (2.5 mL) was added dropwise at room temperature, and the reaction was continued with stirring for 3 hours.
过滤得到510毫克白色固体(S)-4-(2-氨基-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：100％)。LCMS：RT＝3.09min，[M+Na] +＝501.19。 Filter to obtain 510 mg of white solid (S)-4-(2-amino-3-(4-(3-(cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenyl)propionamido group ) Tert-butyl benzoate (yield: 100%). LCMS: RT = 3.09 min, [M+Na] + =501.19.
步骤D：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(3-(cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenylpropionamido)benzoate
Figure PCTCN2020133493-appb-000280
Figure PCTCN2020133493-appb-000280
将2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(268毫克，1.0毫摩尔)，(S)-4-(2-氨基-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(510毫克，1.0毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(570毫克，1.5毫摩尔)和N,N-二异丙基乙胺(523微升，3.0毫摩尔)溶解于N,N-二甲基甲酰胺(5毫升)，室温搅拌过夜。Add 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (268 mg, 1.0 mmol), (S)-4-(2-amino Tert-Butyl-3-(4-(3-(cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenyl)propionamido)benzoate (510 mg, 1.0 mmol), 2 -(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (570 mg, 1.5 mmol) and N,N-diisopropylethylamine (523 μl, 3.0 mmol) was dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature 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410毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基丙酰胺基)苯甲酸叔丁酯(收率：56.3％)。LCMS：RT＝4.37min，[M-H] -＝725.99。 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 410 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) Tert-Butyl-3-(4-(3-(cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenylpropionamido)benzoate (yield: 56.3%). LCMS: RT = 4.37 min, [MH] - = 725.99.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000281
Figure PCTCN2020133493-appb-000281
将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基丙 酰胺基)苯甲酸叔丁酯(410毫克)溶解于三氟乙酸(1.5毫升)和二氯甲烷(7.5毫升)的混合溶剂，室温搅拌1.5小时。(S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(3-(Cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenylpropionamido) tert-butyl benzoate (410 mg) was dissolved in trifluoroacetic acid (1.5 ml) and two A mixed solvent of methyl chloride (7.5 mL) was stirred at room temperature for 1.5 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备高效液相分离。得到50毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基甲基)-2-氧代咪唑啉-1-基)苯基丙酰胺基)苯甲酸(收率：13.2％)。LCMS：RT＝3.85min，[M+Na] +＝694.14。 1H NMR(400MHz,DMSO)δ10.72(s,1H),10.48(s,1H),9.82(s,1H),8.93(d,J＝8.4Hz,1H),7.97(d,J＝2.3Hz,1H),7.92(d,J＝8.8Hz,2H),7.77(d,J＝8.6Hz,1H),7.70(d,J＝8.8Hz,2H),7.62(dd,J＝8.6,2.4Hz,1H),7.48(d,J＝8.7Hz,2H),7.21(d,J＝8.7Hz,2H),4.69(dd,J＝14.5,7.9Hz,1H),3.79(dd,J＝11.3,7.6Hz,2H),3.53(t,J＝8.0Hz,2H),3.10(d,J＝7.5Hz,2H),3.04(d,J＝7.0Hz,2H),0.97-0.87(m,1H),0.52–0.44(m,2H),0.23–0.17(m,2H)。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated by a preparation high performance liquid phase. Obtain 50 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(3-(cyclopropylmethyl)-2-oxoimidazolin-1-yl)phenylpropionamido)benzoic acid (yield: 13.2%). LCMS: RT = 3.85 min , [M+Na] + =694.14. 1 H NMR (400MHz, DMSO) δ 10.72 (s, 1H), 10.48 (s, 1H), 9.82 (s, 1H), 8.93 (d, J = 8.4 Hz, 1H),7.97(d,J=2.3Hz,1H),7.92(d,J=8.8Hz,2H),7.77(d,J=8.6Hz,1H),7.70(d,J=8.8Hz,2H) ,7.62(dd,J=8.6,2.4Hz,1H),7.48(d,J=8.7Hz,2H), 7.21(d,J=8.7Hz,2H), 4.69(dd,J=14.5,7.9Hz, 1H), 3.79 (dd, J = 11.3, 7.6 Hz, 2H), 3.53 (t, J = 8.0 Hz, 2H), 3.10 (d, J = 7.5 Hz, 2H), 3.04 (d, J = 7.0 Hz, 2H), 0.97-0.87 (m, 1H), 0.52-0.44 (m, 2H), 0.23-0.17 (m, 2H).
实施例46Example 46
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]噻吩-2-羧酸Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) benzo[b]thiophene-2-carboxylic acid
Figure PCTCN2020133493-appb-000282
Figure PCTCN2020133493-appb-000282
步骤A：合成(S)-5-(2-氨基-3-苯基丙酰胺基)苯并[b]噻吩-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-amino-3-phenylpropionamido)benzo[b]thiophene-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000283
Figure PCTCN2020133493-appb-000283
将(((9H-芴-9-基)甲氧基)羰基)-L-苯丙氨酸(280毫克，0.72毫摩尔)，5-氨基苯并[b]噻吩-2-羧酸叔丁酯(150毫克，0.60毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(342毫克，0.90毫摩尔)和N,N-二异丙基乙胺(210微升，1.20毫摩尔)溶解于N,N-二甲基甲酰胺(3毫升)，室温搅拌过夜。(((9H-Fluoren-9-yl)methoxy)carbonyl)-L-phenylalanine (280 mg, 0.72 mmol), 5-aminobenzo[b]thiophene-2-carboxylic acid tert-butyl Ester (150 mg, 0.60 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (342 mg, 0.90 mmol) And N,N-diisopropylethylamine (210 μl, 1.20 mmol) were dissolved in N,N-dimethylformamide (3 mL), and stirred at room temperature 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，所得粗品溶解于4-甲基哌啶(1毫升)和二氯甲烷(4毫升)混合溶剂，室温搅拌反应3小时。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 resulting crude product was dissolved in 4-methyl A mixed solvent of piperidine (1 mL) and dichloromethane (4 mL) was stirred and reacted at room temperature for 3 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，所得粗品用硅胶柱层析纯化(石油醚：乙酸乙酯＝1/2)。得到650毫克白色固体(S)-5-(2-氨基-3-苯基丙酰胺基)苯并[b]噻吩-2-羧酸叔丁酯(收率：两步100％)。LCMS：RT＝3.14min，[M+H] +＝397.16。 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 = 1/2). 650 mg of white solid (S)-5-(2-amino-3-phenylpropionamido)benzo[b]thiophene-2-carboxylic acid tert-butyl ester was obtained (yield: 100% in two steps). LCMS: RT = 3.14 min, [M+H] + =397.16.
步骤B：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]噻吩-2-羧酸叔丁酯Step B: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-Phenylpropionamido)benzo[b]thiophene-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000284
Figure PCTCN2020133493-appb-000284
将2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(277毫克，1.03毫摩尔)，(S)-5-(2-氨基-3-苯基丙酰胺基)苯并[b]噻吩-2-羧酸叔丁酯(450毫克，1.14毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(590毫克，1.55毫摩尔)和N,N-二异丙基乙胺(397微升，2.28毫摩尔)溶解于N,N-二甲基甲酰胺(5毫升)，室温搅拌过夜。Add 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (277 mg, 1.03 mmol), (S)-5-(2-amino -3-phenylpropionamido) benzo[b]thiophene-2-carboxylic acid tert-butyl ester (450 mg, 1.14 mmol), 2-(7-azobenzotriazole)-N,N, N',N'-tetramethylurea hexafluorophosphate (590 mg, 1.55 mmol) and N,N-diisopropylethylamine (397 μl, 2.28 mmol) were dissolved in N,N-dimethyl Methyl formamide (5 mL), stirred at room temperature 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340毫克(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]噻吩-2-羧酸叔丁酯(收率：50.9％)。LCMS：RT＝4.54min，[M+H] +＝646.10。 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 340 mg of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)benzo[b]thiophene-2-carboxylic acid tert-butyl ester (yield: 50.9%). LCMS: RT = 4.54 min, [M+H] + =646.10.
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]噻吩-2-羧酸Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-phenylpropionamido)benzo[b]thiophene-2-carboxylic acid
Figure PCTCN2020133493-appb-000285
Figure PCTCN2020133493-appb-000285
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]噻吩-2-羧酸叔丁酯(340毫克，0.53毫摩尔)溶解于三氟乙酸(1.5毫升)和二氯甲烷(7.5毫升)的混合溶剂，室温搅拌3小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene 4-methylpropionamido)benzo[b]thiophene-2-carboxylic acid tert-butyl ester (340 mg, 0.53 mmol) was dissolved in a mixed solvent of trifluoroacetic acid (1.5 mL) and dichloromethane (7.5 mL), and stirred at room temperature 3 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备高效液相分离。得到50毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]噻吩-2-羧酸(收率：16.1％)。LCMS：RT＝4.54min，[M+H] +＝589.96。 1H NMR(400MHz,DMSO)δ13.61–13.31(m,1H),10.72(s,1H),10.36(s,1H),9.82(s,1H),8.95(d,J＝8.3Hz,1H),8.31(d,J＝2.0Hz,1H),8.11(d,J＝0.5Hz,1H),7.99(d,J＝8.8Hz,1H),7.97(d,J＝2.3Hz,1H),7.77(d,J＝8.6Hz,1H),7.61(ddd,J＝8.7,6.2,2.2Hz,2H),7.33–7.24(m,4H),7.24–7.18(m,1H),4.73(dd,J＝14.5,8.0Hz,1H),3.17(d,J＝7.7Hz,2H)。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated by a preparation high performance liquid phase. Obtain 50 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-phenylpropionamido)benzo[b]thiophene-2-carboxylic acid (yield: 16.1%). LCMS: RT = 4.54 min, [M+H] + = 589.96. 1 H NMR (400MHz, DMSO)δ13.61–13.31(m,1H), 10.72(s,1H), 10.36(s,1H), 9.82(s,1H), 8.95(d,J=8.3Hz,1H), 8.31(d, J = 2.0Hz, 1H), 8.11 (d, J = 0.5 Hz, 1H), 7.99 (d, J = 8.8 Hz, 1H), 7.97 (d, J = 2.3 Hz, 1H), 7.77 (d, J = 8.6Hz, 1H), 7.61 (ddd, J = 8.7, 6.2, 2.2 Hz, 2H), 7.33-7.24 (m, 4H), 7.24-7.18 (m, 1H), 4.73 (dd, J = 14.5, 8.0 Hz , 1H), 3.17 (d, J=7.7 Hz, 2H).
实施例47Example 47
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]呋喃-2-羧酸Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) benzo[b]furan-2-carboxylic acid
Figure PCTCN2020133493-appb-000286
Figure PCTCN2020133493-appb-000286
步骤A：合成(S)-5-(2-氨基-3-苯基丙酰胺基)苯并[b]呋喃-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-amino-3-phenylpropionamido)benzo[b]furan-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000287
Figure PCTCN2020133493-appb-000287
将(((9H-芴-9-基)甲氧基)羰基)-L-苯丙氨酸(439毫克，1.133毫摩尔)，5-氨基苯并[b]噻吩-2-羧酸叔丁酯(220毫克，0.944毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(539毫克，1.416毫摩尔)和N,N-二异丙基乙胺(329微升，1.888毫摩尔)溶解于N,N-二甲基甲酰胺(5毫升)，室温搅拌过夜。(((9H-Fluoren-9-yl)methoxy)carbonyl)-L-phenylalanine (439 mg, 1.133 mmol), 5-aminobenzo[b]thiophene-2-carboxylic acid tert-butyl Ester (220 mg, 0.944 mmol), 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (539 mg, 1.416 mmol) And N,N-diisopropylethylamine (329 microliters, 1.888 mmol) were dissolved in N,N-dimethylformamide (5 mL), and stirred at room temperature 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，所得粗品溶解于4-甲基哌啶(1毫升)和二氯甲烷(4毫升)混合溶剂，室温搅拌反应3小时。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 resulting crude product was dissolved in 4-methyl A mixed solvent of piperidine (1 mL) and dichloromethane (4 mL) was stirred and reacted at room temperature for 3 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，所得粗品用硅胶柱层析纯化(石油醚：乙酸乙酯＝1/2)。得到650毫克白色固体(S)-5-(2-氨基-3-苯基丙酰胺基)苯并[b]呋喃-2-羧酸叔丁酯(收率：两步100％)。LCMS：RT＝3.01min，[M-H] -＝379.23。 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 = 1/2). 650 mg of white solid (S)-5-(2-amino-3-phenylpropionamido)benzo[b]furan-2-carboxylic acid tert-butyl ester was obtained (yield: 100% in two steps). LCMS: RT = 3.01 min, [MH] - =379.23.
步骤B：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]呋喃-2-羧酸叔丁酯Step B: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-phenylpropionamido)benzo[b]furan-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000288
Figure PCTCN2020133493-appb-000288
将2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(353毫克，1.31毫摩尔)，(S)-5-(2-氨基-3-苯基丙酰胺基)苯并[b]呋喃-2-羧酸叔丁酯(650毫克，1.71毫摩尔)，2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(742毫克，1.95毫摩尔)和N,N-二异丙基乙胺(457微升，2.62毫摩尔)溶解于N,N-二甲基甲酰胺(5毫升)，室温搅拌过夜。Add 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (353 mg, 1.31 mmol), (S)-5-(2-amino -3-Phenylpropionamido)benzo[b]furan-2-carboxylic acid tert-butyl ester (650 mg, 1.71 mmol), 2-(7-azobenzotriazole)-N,N, N',N'-tetramethylurea hexafluorophosphate (742 mg, 1.95 mmol) and N,N-diisopropylethylamine (457 μl, 2.62 mmol) were dissolved in N,N-dimethyl Methyl formamide (5 mL), stirred at room temperature 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100毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]呋喃-2-羧酸叔丁酯(收率：12.1％)。LCMS：RT＝4.35min，[M+H] +＝630.10。 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 100 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-Phenylpropionamido)benzo[b]furan-2-carboxylic acid tert-butyl ester (yield: 12.1%). LCMS: RT = 4.35 min, [M+H] + =630.10.
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]呋喃-2-羧酸Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-phenylpropionamido)benzo[b]furan-2-carboxylic acid
Figure PCTCN2020133493-appb-000289
Figure PCTCN2020133493-appb-000289
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]呋喃-2-羧酸叔丁酯(100毫克，0.16毫摩尔)溶解于三氟乙酸(1毫升)和二氯甲烷(5毫升)的混合溶剂，室温搅拌3小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene 4-methylpropionamido)benzo[b]furan-2-carboxylic acid tert-butyl ester (100 mg, 0.16 mmol) was dissolved in a mixed solvent of trifluoroacetic acid (1 mL) and dichloromethane (5 mL), and stirred at room temperature 3 hours.
将反应液用旋转蒸发仪浓缩，粗品用制备高效液相分离。得到20毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯并[b]呋喃-2-羧酸(收率：22.0％)。LCMS：RT＝3.97min，[M+Na] +＝596.04。 1H NMR(400MHz,DMSO)δ13.56(s,1H),10.72(s,1H),10.30(s,1H),9.82(s,1H),8.92(d,J＝8.3Hz,1H),8.12(d,J＝2.0Hz,1H),7.97(d,J＝2.3Hz,1H),7.77(d,J＝8.6Hz,1H),7.71–7.65(m,2H),7.62(dd,J＝8.6,2.4Hz,1H),7.53(dd,J＝9.0,2.1Hz,1H),7.29(dd,J＝7.9,5.5Hz,3H),7.24-7.19(m,1H),4.72(dd,J＝14.5,7.9Hz,1H),3.17(d,J＝7.3Hz,2H)。 The reaction solution was concentrated with a rotary evaporator, and the crude product was separated by a preparation high performance liquid phase. Obtain 20 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-phenylpropionamido)benzo[b]furan-2-carboxylic acid (yield: 22.0%). LCMS: RT = 3.97 min, [M+Na] + =596.04. 1 H NMR (400MHz, DMSO)δ13.56(s,1H), 10.72(s,1H), 10.30(s,1H), 9.82(s,1H), 8.92(d,J=8.3Hz,1H), 8.12(d,J= 2.0Hz, 1H), 7.97 (d, J = 2.3 Hz, 1H), 7.77 (d, J = 8.6 Hz, 1H), 7.71-7.65 (m, 2H), 7.62 (dd, J = 8.6, 2.4 Hz, 1H), 7.53 (dd, J = 9.0, 2.1 Hz, 1H), 7.29 (dd, J = 7.9, 5.5 Hz, 3H), 7.24-7.19 (m, 1H), 4.72 (dd, J = 14.5, 7.9 Hz , 1H), 3.17 (d, J=7.3 Hz, 2H).
实施例48Example 48
合成(S)-4-(2-(2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropane Amido) benzoic acid
Figure PCTCN2020133493-appb-000290
Figure PCTCN2020133493-appb-000290
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-氯-2-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯胺Step A: Synthesis of 5-chloro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline
Figure PCTCN2020133493-appb-000291
Figure PCTCN2020133493-appb-000291
将2-溴-5-氯苯胺(4.10克,20.0毫摩尔),4,4,4'，4'，5,5,5'，5'-八甲基-2,2'-(1,3,2-二氧杂硼烷)(9.2克,36.2毫摩尔)和醋酸钾(3.9克,39.8毫摩尔)溶于二甲基亚砜(30.0毫升)中。氮气保护后,加[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(59毫克,0.07毫摩尔),置换氮气,加热升温到80℃并恒温搅拌2.5小时后,LCMS监测至反应完全,将反应液冷却至室温。The 2-bromo-5-chloroaniline (4.10 g, 20.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-(1, 3,2-Dioxaborane) (9.2 g, 36.2 mmol) and potassium acetate (3.9 g, 39.8 mmol) were dissolved in dimethyl sulfoxide (30.0 mL). After nitrogen protection, add [1,1'-bis(diphenylphosphine)ferrocene] palladium dichloride dichloromethane complex (59 mg, 0.07 mmol), replace the nitrogen, heat up to 80 ℃ and After stirring for 2.5 hours at constant temperature, LCMS monitored until the reaction was complete, and the reaction solution was cooled to room temperature.
将反应液缓慢滴加到水(150.0毫升)中,混合液用乙酸乙酯(60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：乙酸乙酯/石油醚＝1/20)得到5.0克黄色固体5-氯-2-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯胺(收率：98.4％)。LCMS：RT＝4.38min,[M+H] +＝254.15。 The reaction solution was slowly added dropwise to water (150.0 ml), the mixed solution was extracted with ethyl acetate (60 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (30 ml × 3 times), anhydrous sulfuric acid Dry with sodium and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/20) to obtain 5.0 g of yellow solid 5-chloro-2-(4,4,5,5-tetramethyl-1 ,3,2-Dioxaborolan-2-yl)aniline (yield: 98.4%). LCMS: RT = 4.38 min, [M+H] + =254.15.
步骤B：合成5-氯-2-(1H-咪唑-2-基)苯胺Step B: Synthesis of 5-chloro-2-(1H-imidazol-2-yl)aniline
Figure PCTCN2020133493-appb-000292
Figure PCTCN2020133493-appb-000292
将5-氯-2-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)苯胺(1.0克,4.1毫摩尔)和2-溴-1H-咪唑(882.0毫克,6.1毫摩尔)溶于1,4-二氧六环(12.0毫升)中，然后将碳酸铯(2.7克,8.0毫摩尔)溶于水(3.0毫升)加入反应液中,氮气保护后,四三苯基膦钯(462.0毫克,0.4毫摩尔),在70℃下搅拌过夜,LCMS监测至反应完全。Combine 5-chloro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (1.0 g, 4.1 mmol) and 2-bromo- 1H-imidazole (882.0 mg, 6.1 mmol) was dissolved in 1,4-dioxane (12.0 ml), and then cesium carbonate (2.7 g, 8.0 mmol) was dissolved in water (3.0 ml) and added to the reaction mixture After nitrogen protection, tetrakistriphenylphosphine palladium (462.0 mg, 0.4 mmol) was stirred at 70°C overnight, and LCMS monitored until the reaction was complete.
反应液冷却至室温,垫硅藻土过滤,滤饼用乙酸乙酯(10毫升×3次)洗涤,合并滤液及洗涤液,有机相经水和饱和食盐水 各洗一次,减压浓缩，残渣经硅胶柱层析纯化得到250毫克黄色固体5-氯-2-(1H-咪唑-2-基)苯胺(收率：32.0％)。LCMS：RT＝2.84min,[M+H] +＝213.12。 The reaction solution was cooled to room temperature, filtered through a pad of celite, the filter cake was washed with ethyl acetate (10 ml×3 times), the filtrate and washing liquid were combined, the organic phase was washed once with water and saturated brine, and concentrated under reduced pressure. Purified by silica gel column chromatography to obtain 250 mg of yellow solid 5-chloro-2-(1H-imidazol-2-yl)aniline (yield: 32.0%). LCMS: RT = 2.84 min, [M+H] + =213.12.
步骤C：合成2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000293
Figure PCTCN2020133493-appb-000293
氮气保护下,将5-氯-2-(1H-咪唑-2-基)苯胺(580毫克,3.0毫摩尔)和吡啶(0.6毫升,9.0毫摩尔)溶于二氯甲烷(3.0毫升)中,冰水浴中搅拌5分钟,向反应液中缓慢滴加草酰氯单甲酯(442毫克,3.5毫摩尔)的二氯甲烷溶液(3.0毫升),冰水浴中搅拌1小时。Under nitrogen protection, dissolve 5-chloro-2-(1H-imidazol-2-yl)aniline (580 mg, 3.0 mmol) and pyridine (0.6 mL, 9.0 mmol) in dichloromethane (3.0 mL), Stir in an ice-water bath for 5 minutes, slowly add dropwise a dichloromethane solution (3.0 ml) of methyl oxalyl chloride (442 mg, 3.5 mmol) to the reaction solution, and stir in an ice-water bath for 1 hour.
向反应液中加水(10.0毫升)淬灭反应,用乙酸乙酯(10毫升×3次)萃取。合并有机相,减压浓缩,所得残余物用硅胶柱层析纯化得到660毫克黄色固体2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酸甲酯(收率：78.0％)。Water (10.0 mL) was added to the reaction solution to quench the reaction, and it was extracted with ethyl acetate (10 mL×3 times). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain 660 mg of yellow solid 2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2-oxy Methyl acetate (yield: 78.0%).
步骤D：合成2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酸Step D: Synthesis of 2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000294
Figure PCTCN2020133493-appb-000294
将2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酸甲酯(660.0毫克,2.4毫摩尔)溶于四氢呋喃(18.0毫升)和水(6.0毫升)中。随后,向上述溶液中加入氢氧化锂一水合物(99毫克,2.37毫摩尔)。在室温下搅拌1小时。Dissolve methyl 2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2-oxoacetate (660.0 mg, 2.4 mmol) in tetrahydrofuran (18.0 ml) and water (6.0 mL). Subsequently, lithium hydroxide monohydrate (99 mg, 2.37 mmol) was added to the above solution. Stir at room temperature for 1 hour.
加稀盐酸调节pH至弱酸性,减压浓缩除去大部分溶剂,加稀盐酸调节pH至3,析出大量白色固体,过滤,收集滤饼得到400毫克黄色固体2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酸(收率：63％)。LCMS：RT＝0.98min,[M+H] +＝266.08。 Add dilute hydrochloric acid to adjust the pH to weak acidity, concentrate under reduced pressure to remove most of the solvent, add dilute hydrochloric acid to adjust the pH to 3, precipitate a large amount of white solid, filter, collect the filter cake to obtain 400 mg of yellow solid 2-((5-chloro-2- (1H-imidazol-2-yl)phenyl)amino)-2-oxoacetic acid (Yield: 63%). LCMS: RT=0.98 min, [M+H] + =266.08.
步骤E：合成(S)-4-(2-(2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2-oxoacetamido)-3- Phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000295
Figure PCTCN2020133493-appb-000295
将2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酸(230毫克,0.98毫摩尔),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(494毫克,1.3毫摩尔)和N,N-二异丙基乙胺(0.2毫升,1.3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(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(220毫克,0.65毫摩尔),在室温下搅拌2小时。The 2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2-oxoacetic acid (230 mg, 0.98 mmol), 2-(7-azobenzotris Nitrozole)-N,N,N',N'-tetramethylurea hexafluorophosphate (494mg, 1.3mmol) and N,N-diisopropylethylamine (0.2ml, 1.3mmol) dissolved In N,N-dimethylformamide (3.0 mL). Subsequently, tert-butyl (S)-4-(2-amino-3-phenylpropionamido)benzoate (220 mg, 0.65 mmol) was added to the above solution and stirred at room temperature for 2 hours.
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得到110毫克黄色固体(S)-4-(2-(2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：31.0％)。LCMS：RT＝4.42min,[M+H] +＝588.21。 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 to obtain 110 mg of yellow solid (S)-4-(2-(2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2 -Oxoacetamido)-3-phenylpropionamido)tert-butyl benzoate (yield: 31.0%). LCMS: RT = 4.42 min, [M+H] + =588.21.
步骤F：合成(S)-4-(2-(2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2-oxoacetamido)-3- Phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000296
Figure PCTCN2020133493-appb-000296
将(S)-4-(2-(2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(110毫克,0.11毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-4-(2-(2-((5-chloro-2-(1H-imidazol-2-yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropane Amido) tert-butyl benzoate (110 mg, 0.11 mmol) was dissolved in dichloromethane (2.0 mL). Subsequently, trifluoroacetic acid (0.5 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7.7毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-咪唑-2-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：8.0％)。LCMS：RT＝3.66min,[M+H] +＝530.20。 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 7.7 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-imidazol-2-yl)phenyl) Amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid (yield: 8.0%). LCMS: RT = 3.66 min, [M+H] + =530.20.
实施例49Example 49
合成(S)-4-(2-(2-(((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000297
Figure PCTCN2020133493-appb-000297
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-氯-3-氟-2-硝基苯胺Step A: Synthesis of 4-chloro-3-fluoro-2-nitroaniline
Figure PCTCN2020133493-appb-000298
Figure PCTCN2020133493-appb-000298
将3-氟-2-硝基苯胺(9.0克,57.7毫摩尔),和N-氯代丁二酰亚胺(15.6克,69.2毫摩尔)溶于乙腈(150.0毫升)中。氮气保护后,加热升温到50℃并恒温搅拌2.5小时后,LCMS监测至反应完全,将反应液冷却至室温。3-Fluoro-2-nitroaniline (9.0 g, 57.7 mmol), and N-chlorosuccinimide (15.6 g, 69.2 mmol) were dissolved in acetonitrile (150.0 mL). After nitrogen protection, heating to 50°C and stirring at constant temperature for 2.5 hours, LCMS monitored until the reaction was complete, and the reaction solution was cooled to room temperature.
将反应液缓慢滴加到水(150.0毫升)中,混合液用乙酸乙酯(60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得到6.8克红棕色固体4-氯-3-氟-2-硝基苯胺(收率：62.3％)。LCMS：RT＝3.66min。The reaction solution was slowly added dropwise to water (150.0 ml), the mixed solution was extracted with ethyl acetate (60 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (30 ml × 3 times), anhydrous sulfuric acid Dry with sodium and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 6.8 g of red-brown solid 4-chloro-3-fluoro-2-nitroaniline (yield: 62.3%). LCMS: RT = 3.66 min.
步骤B：合成2-(4-氯-3-氟-2-硝基苯基)异吲哚啉-1,3-二酮Step B: Synthesis of 2-(4-chloro-3-fluoro-2-nitrophenyl)isoindoline-1,3-dione
Figure PCTCN2020133493-appb-000299
Figure PCTCN2020133493-appb-000299
将4-氯-3-氟-2-硝基苯胺(1.1克,6.0毫摩尔)和异苯并呋喃-1,3-二酮(1.8克,12.1毫摩尔)溶于冰醋酸(20.0毫升)中，氮气保护后,在130℃下搅拌8小时,LCMS监测至反应完全。Dissolve 4-chloro-3-fluoro-2-nitroaniline (1.1 g, 6.0 mmol) and isobenzofuran-1,3-dione (1.8 g, 12.1 mmol) in glacial acetic acid (20.0 mL) After being protected by nitrogen, the mixture was stirred at 130°C for 8 hours, monitored by LCMS until the reaction was complete.
反应液缓慢滴加到冰水中,析出大量固体,过滤,滤饼用水(10毫升×3次)洗涤,滤饼用乙酸乙酯(10毫升)溶解，滴加到正己烷(150毫升),析出大量固体,过滤,滤饼用正己烷(10毫升×3次)洗涤,收集滤饼得到1.4克灰色固体2-(4-氯-3-氟-2-硝基苯基)异吲哚啉-1,3-二酮(收率：32.0％)。LCMS：RT＝4.08min。The reaction solution was slowly added dropwise to ice water, a large amount of solids precipitated, filtered, the filter cake was washed with water (10 ml × 3 times), the filter cake was dissolved with ethyl acetate (10 ml), and added dropwise to n-hexane (150 ml) to precipitate A large amount of solids were filtered, the filter cake was washed with n-hexane (10 ml × 3 times), and the filter cake was collected to obtain 1.4 g of gray solid 2-(4-chloro-3-fluoro-2-nitrophenyl)isoindoline- 1,3-Dione (yield: 32.0%). LCMS: RT = 4.08 min.
步骤C：合成2-(2-氨基-4-氯-3-氟苯基)异吲哚啉-1,3-二酮Step C: Synthesis of 2-(2-amino-4-chloro-3-fluorophenyl)isoindoline-1,3-dione
Figure PCTCN2020133493-appb-000300
Figure PCTCN2020133493-appb-000300
将2-(4-氯-3-氟-2-硝基苯基)异吲哚啉-1,3-二酮(740毫克,2.3毫摩尔)溶于甲醇(30.0毫升)和冰醋酸(2.0毫升)中。随后,向上述溶液中加入还原性铁粉(1.3克,23.0毫摩尔),在65℃下搅拌2小时。Dissolve 2-(4-chloro-3-fluoro-2-nitrophenyl)isoindoline-1,3-dione (740 mg, 2.3 mmol) in methanol (30.0 mL) and glacial acetic acid (2.0 Milliliters). Subsequently, reducing iron powder (1.3 g, 23.0 mmol) was added to the above solution and stirred at 65°C for 2 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,所得残余物用硅胶柱层析纯化得到205毫克黄色固体2-(2-氨基-4-氯-3-氟苯基)异吲哚啉-1,3-二酮(收率54.0％)。LCMS：RT＝3.83min,[M+H] +＝291.08。 The reaction solution was cooled to room temperature. Add sodium bicarbonate to neutralize, and extract the mixture with ethyl acetate (20 ml×3 times). The organic phases were combined, and the organic phase was first dried with saturated brine (10 ml × 3 times), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue obtained was purified by silica gel column chromatography to obtain 205 mg of yellow solid 2-(2-amino- 4-chloro-3-fluorophenyl)isoindoline-1,3-dione (yield 54.0%). LCMS: RT = 3.83 min, [M+H] + =291.08.
步骤D：合成2-(4-氯-2-(二烯丙基氨基)-3-氟苯基)异吲哚啉-1,3-二酮Step D: Synthesis of 2-(4-chloro-2-(diallylamino)-3-fluorophenyl)isoindoline-1,3-dione
Figure PCTCN2020133493-appb-000301
Figure PCTCN2020133493-appb-000301
将2-(2-氨基-4-氯-3-氟苯基)异吲哚啉-1,3-二酮(1.4克,8.4毫摩尔)和N,N-二异丙基乙胺(6.5毫升,42.0毫摩尔)溶于N,N-二甲基甲酰胺(10.0毫升)中。随后,向上述溶液中加入3-溴丙-1-烯(3.2毫升,33.6毫摩尔)。在90℃下搅拌10小时。Combine 2-(2-amino-4-chloro-3-fluorophenyl)isoindoline-1,3-dione (1.4 g, 8.4 mmol) and N,N-diisopropylethylamine (6.5 ML, 42.0 mmol) was dissolved in N,N-dimethylformamide (10.0 mL). Subsequently, 3-bromoprop-1-ene (3.2 mL, 33.6 mmol) was added to the above solution. Stir at 90°C for 10 hours.
向反应液中加水(30.0毫升)淬灭反应,用乙酸乙酯(20毫升×3次)萃取。合并有机相,减压浓缩,所得残余物用硅胶柱层析纯化得到1.4克黄色油状物2-(4-氯-2-(二烯丙基氨基)-3-氟苯基)异吲哚啉-1,3-二酮(收率：86％)。LCMS：RT＝4.69min,[M+H] +＝371.12。 Water (30.0 mL) was added to the reaction solution to quench the reaction, and it was extracted with ethyl acetate (20 mL×3 times). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain 1.4 g of yellow oil 2-(4-chloro-2-(diallylamino)-3-fluorophenyl)isoindoline -1,3-dione (yield: 86%). LCMS: RT = 4.69 min, [M+H] + = 371.12.
步骤E：合成N1,N1-二烯丙基-5-氯-6-氟苯-1,2-二胺Step E: Synthesis of N1,N1-diallyl-5-chloro-6-fluorobenzene-1,2-diamine
Figure PCTCN2020133493-appb-000302
Figure PCTCN2020133493-appb-000302
将2-(4-氯-2-(二烯丙基氨基)-3-氟苯基)异吲哚啉-1,3-二酮(1.4克,3.9毫摩尔)溶于乙醇(20.0毫升)中。随后,向上述溶液中加入水合肼(1.9毫升,39.2毫摩尔)。在85℃下搅拌2小时。Dissolve 2-(4-chloro-2-(diallylamino)-3-fluorophenyl)isoindoline-1,3-dione (1.4g, 3.9mmol) in ethanol (20.0ml) in. Subsequently, hydrazine hydrate (1.9 mL, 39.2 mmol) was added to the above solution. Stir at 85°C for 2 hours.
反应液趁热过滤,滤液经减压浓缩,所得残余物用硅胶柱层析纯化得到990毫克黄色固体N1，N1-二烯丙基-5-氯-6-氟苯-1,2-二胺(收率：86％)。LCMS：RT＝4.46min,[M+H] +＝241.11。 The reaction solution was filtered while hot, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 990 mg of yellow solid N1,N1-diallyl-5-chloro-6-fluorobenzene-1,2-diamine (Yield: 86%). LCMS: RT = 4.46 min, [M+H] + =241.11.
步骤F：合成N,N-二烯丙基-3-氯-2-氟-6-(1H-四唑-1-基)苯胺Step F: Synthesis of N,N-diallyl-3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)aniline
Figure PCTCN2020133493-appb-000303
Figure PCTCN2020133493-appb-000303
将N1，N1-二烯丙基-5-氯-6-氟苯-1,2-二胺(760毫克,3.2毫摩尔)溶于冰醋酸(4.0毫升)中。氮气保护后,冰水浴中搅 拌15分钟,加入原甲酸三甲酯(1.6毫升,13.9毫摩尔),搅拌15分钟后,加入叠氮化钠(824毫克,12.7毫摩尔),氮气保护后,在85℃下搅拌16小时。Dissolve N1,N1-diallyl-5-chloro-6-fluorobenzene-1,2-diamine (760 mg, 3.2 mmol) in glacial acetic acid (4.0 mL). After nitrogen protection, stir in an ice water bath for 15 minutes, add trimethyl orthoformate (1.6 ml, 13.9 mmol), stir for 15 minutes, add sodium azide (824 mg, 12.7 mmol), after nitrogen protection, Stir at 85°C for 16 hours.
将反应液缓慢滴加到水(20.0毫升)中,过滤,滤液用亚硝酸钠处理后倒入废液,收集滤饼得到800毫克黄色固体N，N-二烯丙基-3-氯-2-氟-6-(1H-四唑-1-基)苯胺(收率：88％)。LCMS：RT＝4.29min,[M+Na] +＝316.97。 The reaction solution was slowly added dropwise to water (20.0 ml), filtered, and the filtrate was treated with sodium nitrite and poured into the waste liquid. The filter cake was collected to obtain 800 mg of yellow solid N, N-diallyl-3-chloro-2 -Fluoro-6-(1H-tetrazol-1-yl)aniline (yield: 88%). LCMS: RT = 4.29 min, [M+Na] + =316.97.
步骤G：合成3-氯-2-氟-6-(1H-四唑-1-基)苯胺Step G: Synthesis of 3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)aniline
Figure PCTCN2020133493-appb-000304
Figure PCTCN2020133493-appb-000304
将N，N-二烯丙基-3-氯-2-氟-6-(1H-四唑-1-基)苯胺(800毫克,2.7毫摩尔)和1,3-二甲基巴比妥酸(2.1克,13.7毫摩尔)溶于二氯甲烷(20.0毫升)中。氮气保护后,加入四三苯基膦钯(120毫克,0.1毫摩尔),在40℃下搅拌2小时。Combine N,N-diallyl-3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)aniline (800 mg, 2.7 mmol) and 1,3-dimethylbarbital The acid (2.1 g, 13.7 mmol) was dissolved in dichloromethane (20.0 mL). After nitrogen protection, tetrakistriphenylphosphine palladium (120 mg, 0.1 mmol) was added and stirred at 40°C for 2 hours.
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得到700毫克黄色固体3-氯-2-氟-6-(1H-四唑-1-基)苯胺(收率：96％)。LCMS：RT＝3.45min,[M-H] -＝214.01。 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 to obtain 700 mg of yellow solid 3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)aniline (yield: 96%). LCMS: RT = 3.45 min, [MH] - =214.01.
步骤H：合成2-((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸甲酯Step H: Synthesis of methyl 2-((3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000305
Figure PCTCN2020133493-appb-000305
氮气保护下,将3-氯-2-氟-6-(1H-四唑-1-基)苯胺(700毫克,3.3毫摩尔)和吡啶(0.6毫升,9.0毫摩尔)溶于二氯甲烷(10.0毫升)中,冰水浴中搅拌5分钟,向反应液中缓慢滴加草酰氯单甲酯(480毫克,3.5毫摩尔)的二氯甲烷溶液(3.0毫升),冰水浴中搅拌1小时。Under nitrogen protection, 3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)aniline (700 mg, 3.3 mmol) and pyridine (0.6 mL, 9.0 mmol) were dissolved in dichloromethane ( 10.0 ml), stirred in an ice-water bath for 5 minutes, slowly added dropwise a dichloromethane solution (3.0 ml) of monomethyl oxalyl chloride (480 mg, 3.5 mmol) to the reaction solution, and stirred in an ice-water bath for 1 hour.
向反应液中加水(10.0毫升)淬灭反应,用乙酸乙酯(10毫升×3次)萃取。合并有机相,减压浓缩,所得残余物用硅胶柱层析纯化得到860毫克黄色固体2-((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸甲酯(收率：87.0％)。LCMS：RT＝3.21min,[M+H] +＝300.07。 Water (10.0 mL) was added to the reaction solution to quench the reaction, and it was extracted with ethyl acetate (10 mL×3 times). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain 860 mg of yellow solid 2-((3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)phenyl)amino ) Methyl-2-oxoacetate (yield: 87.0%). LCMS: RT=3.21 min, [M+H] + =300.07.
步骤I：合成2-((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸Step I: Synthesis of 2-((3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000306
Figure PCTCN2020133493-appb-000306
将2-((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸甲酯(860.0毫克,2.9毫摩尔)溶于四氢呋喃(18.0毫升)和水(6.0毫升)中。随后,向上述溶液中加入氢氧化锂一水合物(120毫克,2.9毫摩尔)。在室温下搅拌1小时。Methyl 2-((3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetate (860.0 mg, 2.9 mmol) was dissolved in tetrahydrofuran ( 18.0 ml) and water (6.0 ml). Subsequently, lithium hydroxide monohydrate (120 mg, 2.9 mmol) was added to the above solution. Stir at room temperature for 1 hour.
加稀盐酸调节pH至弱酸性,减压浓缩除去大部分溶剂,加稀盐酸调节pH至3,析出大量白色固体,过滤,收集滤饼得到520毫克黄色固体2-((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(收率：63％)。LCMS：RT＝3.14min,[M-H] -＝285.13。 Add dilute hydrochloric acid to adjust the pH to weak acidity, concentrate under reduced pressure to remove most of the solvent, add dilute hydrochloric acid to adjust the pH to 3, precipitate a large amount of white solid, filter, collect the filter cake to obtain 520 mg of yellow solid 2-((3-chloro-2- Fluoro-6-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (Yield: 63%). LCMS: RT = 3.14 min, [MH] - =285.13.
步骤J：合成叔丁基(S)-4-(2-(2-(((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸酯Step J: Synthesis of tert-butyl (S)-4-(2-(2-(((3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)phenyl)amino)-2- Oxoacetamido)-3-phenylpropionamido)benzoate
Figure PCTCN2020133493-appb-000307
Figure PCTCN2020133493-appb-000307
将2-((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酸(230毫克,0.81毫摩尔),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(462毫克,1.2毫摩尔)和N,N-二异丙基乙胺(0.4毫升,2.4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(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(330毫克,0.97毫摩尔),在室温下搅拌2小时。The 2-((3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (230 mg, 0.81 mmol), 2-(7- Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (462mg, 1.2mmol) and N,N-diisopropylethylamine (0.4ml, 2.4 mmol) was dissolved in N,N-dimethylformamide (3.0 mL). Subsequently, tert-butyl (S)-4-(2-amino-3-phenylpropionamido)benzoate (330 mg, 0.97 mmol) was added to the above solution, and stirred at room temperature for 2 hours.
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得到220毫克黄色固体叔丁基(S)-4-(2-(2-(((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸酯(收率：45.0％)。LCMS：RT＝4.27min,[M-H] -＝606.10。 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 to obtain 220 mg of yellow solid tert-butyl(S)-4-(2-(2-(((3-chloro-2-fluoro-6-(1H-tetrazole-1- (Phenyl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoate (yield: 45.0%). LCMS: RT = 4.27 min, [MH] - =606.10 .
步骤K：合成(S)-4-(2-(2-(((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step K: Synthesis of (S)-4-(2-(2-(((3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000308
Figure PCTCN2020133493-appb-000308
将叔丁基(S)-4-(2-(2-(((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸酯(88毫克,0.15毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。Add tert-butyl(S)-4-(2-(2-(((3-chloro-2-fluoro-6-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-phenylpropionamido)benzoate (88 mg, 0.15 mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 mL) was added to the above solution, Stir 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9.3毫克白色固体(S)-4-(2-(2-(((3-氯-2-氟-6-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：12.0％)。LCMS：RT＝3.72min,[M-H] -＝550.11。 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 9.3 mg of white solid (S)-4-(2-(2-(((3-chloro-2-fluoro-6-(1H-tetrazole-1) -Yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid (yield: 12.0%). LCMS: RT = 3.72 min, [MH] - =550.11.
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-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 PCTCN2020133493-appb-000309
Figure PCTCN2020133493-appb-000309
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(2-甲氧基乙氧基)哌啶-1-甲酸叔丁酯Step A: Synthesis of tert-butyl 4-(2-methoxyethoxy)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000310
Figure PCTCN2020133493-appb-000310
将4-羟基哌啶-1-羧酸叔丁酯(402毫克,2.0毫摩尔)溶于乙腈(10.0毫升)中,冰水浴中搅拌5分钟,向反应液中加钠氢(104毫克,2.6毫摩尔),然后室温搅拌1小时。随后,向上述溶液中加入1-溴-2-甲氧基乙烷(550毫克,4.0毫摩尔),在50℃下搅拌5小时。4-Hydroxypiperidine-1-carboxylic acid tert-butyl ester (402 mg, 2.0 mmol) was dissolved in acetonitrile (10.0 ml), stirred in an ice-water bath for 5 minutes, and sodium hydrogen (104 mg, 2.6 Millimoles), and then stirred at room temperature for 1 hour. Subsequently, 1-bromo-2-methoxyethane (550 mg, 4.0 mmol) was added to the above solution and stirred at 50°C for 5 hours.
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得到280毫克无色油状物4-(2-甲氧基乙氧基) 哌啶-1-甲酸叔丁酯(收率：54.0％)。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 to obtain 280 mg of colorless oily tert-butyl 4-(2-methoxyethoxy)piperidine-1-carboxylate (yield: 54.0%).
步骤B：合成4-(2-甲氧基乙氧基)哌啶的三氟乙酸盐Step B: Synthesis of 4-(2-methoxyethoxy)piperidine trifluoroacetate
Figure PCTCN2020133493-appb-000311
Figure PCTCN2020133493-appb-000311
将4-(2-甲氧基乙氧基)哌啶-1-甲酸叔丁酯(280毫克,1.1毫摩尔)溶于二氯甲烷(6.0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-(2-methoxyethoxy)piperidine-1-carboxylate (280 mg, 1.1 mmol) was dissolved in dichloromethane (6.0 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250毫克黄色固体4-(2-甲氧基乙氧基)哌啶的三氟乙酸盐(收率：90.0％)。The reaction solution was concentrated under reduced pressure in an air bath to obtain 250 mg of trifluoroacetic acid salt of 4-(2-methoxyethoxy)piperidine as a yellow solid (yield: 90.0%).
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-甲酰氨基)苯基)丙酰氨基)-1H-吲哚-1,2-二羧酸二叔丁酯Step C: Synthesis of (S)-5-(2-(4-(5-chloro-2-(1H-tetrazol-1-yl)phenyl)-2,3-dioxopiperazin-1-yl )-3-(-4-(4-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000312
Figure PCTCN2020133493-appb-000312
室温下,将二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(120毫克,0.16毫摩尔)和4-(2-甲氧基乙氧基)哌啶的三氟乙酸盐(203毫克,0.96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(0.5毫升,2.6毫摩尔)。在60℃反应12小时。At room temperature, di-tert-butyl(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylate (120 mg, 0.16 mmol) and 4-( The trifluoroacetate salt of 2-methoxyethoxy)piperidine (203 mg, 0.96 mmol) was dissolved in tetrahydrofuran (6.0 ml). Then, N,N-diisopropylethylamine was added to the above solution (0.5ml, 2.6mmol). 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110毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙氧基)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：60％)。LCMS：RT＝3.97min,[M+Na] +＝851.08。 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 110 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(4-(2-Methoxyethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl Ester (yield: 60%). LCMS: RT = 3.97 min, [M+Na] + =851.08.
步骤D：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step D: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-methoxyethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000313
Figure PCTCN2020133493-appb-000313
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙氧基)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,0.12毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(2-Methoxyethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (110 mg , 0.12 mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 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8毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-甲氧基乙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：8.6％)。LCMS：RT＝3.51min,[M+H] +＝773.24。 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 8 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)benzene) Yl)amino)-2-oxoacetamido)-3-(4-(4-(2-methoxyethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H- Indole-2-carboxylic acid (yield: 8.6%). LCMS: RT = 3.51 min, [M+H] + =773.24.
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-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 PCTCN2020133493-appb-000314
Figure PCTCN2020133493-appb-000314
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(吗啉-4-羰基)哌啶-1-甲酸叔丁酯Step A: Synthesis of tert-butyl 4-(morpholine-4-carbonyl)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000315
Figure PCTCN2020133493-appb-000315
将1-(叔丁氧羰基)哌啶-4-羧酸(902毫克,3.9毫摩尔)和和N,N-二异丙基乙胺(1.9毫升,11.8毫摩尔)溶于1,2-二氯乙烷(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(2.2克,5.9毫摩尔)和吗啉(513毫克,5.9毫摩尔)。在室温下搅拌2小时。Dissolve 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (902 mg, 3.9 mmol) and N,N-diisopropylethylamine (1.9 mL, 11.8 mmol) in 1,2- Dichloroethane (3.0 mL). Subsequently, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.2 g, 5.9 mmol) and morpholine were added to the above solution (513 mg, 5.9 mmol). Stir at room temperature for 2 hours.
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得到1.2克黄色固体4-(吗啉-4-羰基)哌啶-1-甲酸叔丁酯(收率：99.0％)。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 to obtain 1.2 g of tert-butyl 4-(morpholine-4-carbonyl)piperidine-1-carboxylate as a yellow solid (yield: 99.0%).
步骤B：合成吗啉代(哌啶-4-基)甲酮的三氟乙酸盐Step B: Synthesis of the trifluoroacetate salt of morpholino(piperidin-4-yl)methanone
Figure PCTCN2020133493-appb-000316
Figure PCTCN2020133493-appb-000316
将4-(吗啉-4-羰基)哌啶-1-甲酸叔丁酯(1.2克,3.9毫摩尔)溶于二氯甲烷(6.0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-(morpholine-4-carbonyl)piperidine-1-carboxylate (1.2 g, 3.9 mmol) was dissolved in dichloromethane (6.0 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1.1克白色固体吗啉代(哌啶-4-基)甲酮的三氟乙酸盐(收率：93.0％)。The reaction solution was concentrated under reduced pressure in an air bath to obtain 1.1 g of white solid morpholino(piperidin-4-yl)methanone trifluoroacetate (yield: 93.0%).
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(吗啉-4-羰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000317
Figure PCTCN2020133493-appb-000317
室温下,将二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(100毫克,0.12毫摩尔)和4-(2-甲氧基乙氧基)哌啶的三氟乙酸盐(100毫克,0.35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(0.2毫升,1.4毫摩尔)。在60℃反应12小时。At room temperature, di-tert-butyl(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylate (100 mg, 0.12 mmol) and 4-( The trifluoroacetate salt of 2-methoxyethoxy)piperidine (100 mg, 0.35 mmol) was dissolved in tetrahydrofuran (6.0 ml). Then, N,N-diisopropylethylamine was added to the above solution (0.2 mL, 1.4 mmol). 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,所得残余物用硅胶柱层析纯化得到59毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(吗啉-4-羰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：53％)。LCMS：RT＝4.26min,[M+H] +＝968.47。 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 The resulting residue was purified by silica gel column chromatography to obtain 59 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino )-2-oxoacetamido)-3-(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1, Di-tert-butyl 2-dicarboxylic acid (yield: 53%). LCMS: RT = 4.26 min, [M+H] + =968.47.
步骤D：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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 D: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000318
Figure PCTCN2020133493-appb-000318
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(吗啉-4-羰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(59毫克,0.06毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (59 mg, 0.06 (Mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 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12毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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：24％)。LCMS：RT＝3.36min,[M-H] -＝810.28。 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 12 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)benzene) (Amino)-2-oxoacetamido)-3-(4-(4-(morpholine-4-carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole -2-carboxylic acid (yield: 24%). LCMS: RT = 3.36 min, [MH] - =810.28.
实施例52Example 52
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基甲酰基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(Dimethylcarbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000319
Figure PCTCN2020133493-appb-000319
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(二甲基氨基甲酰基)哌啶-1-羧酸叔丁酯Step A: Synthesis of tert-butyl 4-(dimethylcarbamoyl)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000320
Figure PCTCN2020133493-appb-000320
将1-(叔丁氧羰基)哌啶-4-羧酸(902毫克,3.9毫摩尔)和和N,N-二异丙基乙胺(1.9毫升,11.8毫摩尔)溶于1,2-二氯乙烷(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(2.2克,5.9毫摩尔)和二甲胺四氢呋喃溶液(4.0毫升,8.0毫摩尔)。在室温下搅拌2小时。Dissolve 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (902 mg, 3.9 mmol) and N,N-diisopropylethylamine (1.9 mL, 11.8 mmol) in 1,2- Dichloroethane (3.0 mL). Subsequently, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.2 g, 5.9 mmol) and dimethyl urea hexafluorophosphate (2.2 g, 5.9 mmol) were added to the above solution. Amine tetrahydrofuran solution (4.0 mL, 8.0 mmol). Stir at room temperature for 2 hours.
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得到1.0克黄色固体4-(二甲基氨基甲酰基)哌啶-1-羧酸叔丁酯(收率：100％)。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 to obtain 1.0 g of tert-butyl 4-(dimethylcarbamoyl)piperidine-1-carboxylate as a yellow solid (yield: 100%).
步骤B：合成N,N-二甲基哌啶-4-羧酰胺的三氟乙酸盐Step B: Synthesis of trifluoroacetate salt of N,N-dimethylpiperidine-4-carboxamide
Figure PCTCN2020133493-appb-000321
Figure PCTCN2020133493-appb-000321
将4-(二甲基氨基甲酰基)哌啶-1-羧酸叔丁酯(1.0克,3.9毫摩尔)溶于二氯甲烷(6.0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-(dimethylcarbamoyl)piperidine-1-carboxylate (1.0 g, 3.9 mmol) was dissolved in dichloromethane (6.0 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N，N-二甲基哌啶-4-羧酰胺的三氟乙酸盐(收率：100％)。The reaction solution was concentrated under reduced pressure in an air bath to obtain 1.0 g of trifluoroacetate of yellow solid N,N-dimethylpiperidine-4-carboxamide (yield: 100%).
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基甲酰基)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 C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(Dimethylcarbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000322
Figure PCTCN2020133493-appb-000322
室温下,将二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(200毫克,0.23毫摩尔)和N，N-二甲基哌啶-4-羧酰胺的三氟乙酸盐(175毫克,0.69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(0.4毫升,2.8毫摩尔)。在60℃反应12小时。At room temperature, di-tert-butyl(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylate (200 mg, 0.23 mmol) and N, N -Dimethylpiperidine-4-carboxamide trifluoroacetate (175 mg, 0.69 mmol) was dissolved in tetrahydrofuran (6.0 ml). Then, N,N-diisopropylethylamine was added to the above solution (0.4ml, 2.8mmol). 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,所得残余物用硅胶柱层析纯化得到120毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基甲酰基)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：56％)。LCMS：RT＝4.28min,[M+H] +＝926.28。 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 The resulting residue was purified by silica gel column chromatography to obtain 120 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino )-2-oxoacetamido)-3-(4-(4-(dimethylcarbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1, Di-tert-butyl 2-dicarboxylic acid (yield: 56%). LCMS: RT = 4.28 min, [M+H] + =926.28.
步骤D：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基甲酰基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step D: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Dimethylcarbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000323
Figure PCTCN2020133493-appb-000323
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基甲酰基)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(120毫克,0.06毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(Dimethylcarbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (120 mg, 0.06 (Mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 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毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基甲酰基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：25％)。LCMS：RT＝3.38min,[M-H] -＝768.22。 1H NMR(400MHz,DMSO)δ12.91(s,1H),11.72(s,1H),10.73(s,1H),10.04(s,1H),9.84(s,1H),8.77(d,J＝8.4Hz,1H),8.41(s,1H),7.99(d,J＝2.3Hz,1H),7.94(d,J＝1.8Hz,1H),7.77(d,J＝8.6Hz,1H),7.61(dd,J＝8.6,2.4Hz,1H),7.41–7.29(m,4H),7.12(d,J＝8.6Hz,2H),7.07–7.04(m,1H),4.67(dd,J＝14.5,7.8Hz,1H),4.10(d,J＝13.2Hz,2H),3.12–3.05(m,2H),3.04(s,3H),2.85(d,J＝11.0Hz,2H),2.81(s,3H),δ2.53(d,J＝1.9Hz,1H),δ1.63(d,J＝10.7Hz,2H),1.45(dd,J＝20.9,12.1Hz,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 white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(4-(dimethylcarbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole- 2-carboxylic acid (yield: 25%). LCMS: RT = 3.38 min, [MH] - = 768.22. 1 H NMR (400MHz, DMSO) δ 12.91 (s, 1H), 11.72 (s, 1H), 10.73(s,1H),10.04(s,1H),9.84(s,1H),8.77(d,J=8.4Hz,1H),8.41(s,1H),7.99(d,J=2.3Hz,1H ), 7.94 (d, J = 1.8 Hz, 1H), 7.77 (d, J = 8.6 Hz, 1H), 7.61 (dd, J = 8.6, 2.4 Hz, 1H), 7.41-7.29 (m, 4H), 7.12 (d,J=8.6Hz,2H),7.07–7.04(m,1H), 4.67(dd,J=14.5,7.8Hz,1H), 4.10(d,J=13.2Hz,2H), 3.12–3.05( m,2H),3.04(s,3H),2.85(d,J=11.0Hz,2H),2.81(s,3H),δ2.53(d,J=1.9Hz,1H),δ1.63(d , J = 10.7Hz, 2H), 1.45 (dd, J = 20.9, 12.1Hz, 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-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 PCTCN2020133493-appb-000324
Figure PCTCN2020133493-appb-000324
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(2-(叔丁氧基)-2-氧代乙氧基)哌啶-1-甲酸叔丁酯Step A: Synthesis of tert-butyl 4-(2-(tert-butoxy)-2-oxoethoxy)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000325
Figure PCTCN2020133493-appb-000325
将4-羟基哌啶-1-羧酸叔丁酯(1.32克,10.0毫摩尔)溶于N,N-二甲基甲酰胺(10.0毫升)中,冰水浴中搅拌5分钟,向反应液中加钠氢(520毫克,13.0毫摩尔),然后室温搅拌1小时。随后,向上述溶液中加入2-溴乙酸叔丁酯(2.9毫升,20.0毫摩尔),在50℃下搅拌5小时。Dissolve 4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (1.32 g, 10.0 mmol) in N,N-dimethylformamide (10.0 ml), stir in an ice-water bath for 5 minutes, and add to the reaction solution Add sodium hydrogen (520 mg, 13.0 mmol), and then stir at room temperature for 1 hour. Subsequently, tert-butyl 2-bromoacetate (2.9 mL, 20.0 mmol) was added to the above solution and stirred at 50°C for 5 hours.
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得到600毫克无色油状物4-(2-(叔丁氧基)-2-氧代乙氧基)哌啶-1-甲酸叔丁酯(收率：19.0％)。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 to obtain 600 mg of colorless oily 4-(2-(tert-butoxy)-2-oxoethoxy)piperidine-1-carboxylic acid tert-butyl ester (Yield: 19.0 %).
步骤B：合成2-(哌啶-4-基氧基)乙酸叔丁酯盐酸盐Step B: Synthesis of tert-butyl 2-(piperidin-4-yloxy)acetate hydrochloride
Figure PCTCN2020133493-appb-000326
Figure PCTCN2020133493-appb-000326
将4-(2-(叔丁氧基)-2-氧代乙氧基)哌啶-1-甲酸叔丁酯(280毫克,1.1毫摩尔)溶于乙酸乙酯(6.0毫升)中。随后,向上述溶液中加入2.0M盐酸的乙酸乙酯溶液(2.0毫升),在室温下搅拌2小时。Tert-butyl 4-(2-(tert-butoxy)-2-oxoethoxy)piperidine-1-carboxylate (280 mg, 1.1 mmol) was dissolved in ethyl acetate (6.0 mL). Subsequently, 2.0M hydrochloric acid in ethyl acetate (2.0 mL) was added to the above solution, and stirred at room temperature for 2 hours.
将反应液减压浓缩,得到250毫克白色固体2-(哌啶-4-基氧基)乙酸叔丁酯盐酸盐(收率：52％)。The reaction solution was concentrated under reduced pressure to obtain 250 mg of white solid tert-butyl 2-(piperidin-4-yloxy)acetate hydrochloride (yield: 52%).
步骤C：合成(S)-5-(3-(4-(4-(2-(叔丁氧基)-2-氧乙氧基)哌啶-1-甲酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step C: Synthesis of (S)-5-(3-(4-(4-(2-(tert-butoxy)-2-oxoethoxy)piperidine-1-carboxamido)phenyl)-2 -(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
Figure PCTCN2020133493-appb-000327
Figure PCTCN2020133493-appb-000327
室温下,将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(170毫克,0.2毫摩尔)和2-(哌啶-4-基氧基)乙酸叔丁酯盐酸盐(200毫克,0.8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(0.5毫升,2.6毫摩尔)。在60℃反应12小时。At room temperature, add (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (170 mg, 0.2 mmol) and 2-(piper Pyridin-4-yloxy)acetate tert-butyl hydrochloride (200 mg, 0.8 mmol) was dissolved in tetrahydrofuran (6.0 ml). Then, to the above solution, N,N-diisopropylethylamine (0.5 ML, 2.6 mmol). 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,所得残余物用硅胶柱层析纯化得到132毫克黄色固体(S)-5-(3-(4-(4-(2-(叔丁氧基)-2-氧乙氧基)哌啶-1-甲酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：67％)。LCMS：RT＝4.62min,[M-H] -＝984.24。 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 The resulting residue was purified by silica gel column chromatography to obtain 132 mg of yellow solid (S)-5-(3-(4-(4-(2-(tert-butoxy)-2-oxoethoxy)piperidine- 1-carboxamido)phenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido) -1H-Indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 67%). LCMS: RT = 4.62 min, [MH] - =984.24.
步骤D：合成(S)-5-(3-(4-(4-(羧基甲氧基)哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸Step D: Synthesis of (S)-5-(3-(4-(4-(carboxymethoxy)piperidine-1-carboxamido)phenyl)-2-(2-((5-chloro-2 -(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000328
Figure PCTCN2020133493-appb-000328
将二叔丁基(S)-5-(3-(4-(4-(2-(叔丁氧基)-2-氧乙氧基)哌啶-1-甲酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(132毫克,0.13毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。Di-tert-butyl (S)-5-(3-(4-(4-(2-(tert-butoxy)-2-oxoethoxy)piperidine-1-carboxamido)phenyl)- 2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-1,2- The dicarboxylic acid ester (132 mg, 0.13 mmol) was dissolved in dichloromethane (2.0 mL). Subsequently, trifluoroacetic acid (0.5 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12毫克黄色固体(S)-5-(3-(4-(4-(羧基甲氧基)哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸(收率：12％)。LCMS：RT＝3.39min,[M+H] +＝773.08。 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 12 mg of yellow solid (S)-5-(3-(4-(4-(carboxymethoxy)piperidine-1-carboxamido)phenyl )-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2- Carboxylic acid (yield: 12%). LCMS: RT = 3.39 min, [M+H] + =773.08.
实施例54Example 54
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(苯基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-(phenylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000329
Figure PCTCN2020133493-appb-000329
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(phenylsulfonyl)ureido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000330
Figure PCTCN2020133493-appb-000330
室温下,将二叔丁基(S)-5-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(160毫克,0.2毫摩尔)和苯磺酰基异氰酸酯(183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(0.5毫升,2.6毫摩尔)。在60℃反应12小时。At room temperature, di-tert-butyl(S)-5-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)propionamido)-1H-indole-1,2-dicarboxylate (160 mg, 0.2 mmol) and benzenesulfonyl isocyanate (183 mg, 1.0 mmol) ) Was dissolved in tetrahydrofuran (6.0 mL). Subsequently, add N,N-diisopropylethylamine (0.5 mL, 2.6 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,所得残余物用硅胶柱层析纯化得到122毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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：59％)。LCMS：RT＝4.06min,[M+H] +＝827.23。 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 The resulting residue was purified by silica gel column chromatography to obtain 122 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino )-2-oxoacetamido)-3-(4-(3-(phenylsulfonyl)ureido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (Yield: 59%). LCMS: RT=4.06 min, [M+H] + =827.23.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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 (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-(phenylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000331
Figure PCTCN2020133493-appb-000331
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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(122毫克,0.15毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(3-(phenylsulfonyl)ureido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (122 mg, 0.15 mmol) dissolved in dichloromethane ( 2.0 mL). Then, trifluoroacetic acid (0.5 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-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(苯基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：23％)。LCMS：RT＝3.65min,[M-H] -＝770.08。 1H NMR(400MHz,DMSO)δ12.93(s,1H),11.73(s,1H),10.71(s,1H),10.04(s,1H),9.82(s,1H),8.80(s,1H),8.01–7.91(m,4H),7.77(d,J＝8.6Hz,1H),7.69(d,J＝7.5Hz,1H),7.66–7.59(m,3H),7.37(d,J＝8.9Hz,1H),7.31(d,J＝1.9Hz,1H),7.24(d,J＝8.6Hz,2H),7.16(d,J＝8.6Hz,2H),7.05(d,J＝1.4Hz,1H),4.65(dd,J＝14.0,8.2Hz,1H),3.12–3.02(m,2H),2.53(d,J＝1.9Hz,1H)。 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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(3-(phenylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield :twenty three%). LCMS: RT = 3.65 min, [MH] - =770.08. 1 H NMR (400MHz, DMSO) δ 12.93 (s, 1H), 11.73 (s, 1H), 10.71 (s, 1H), 10.04 (s, 1H), 9.82 (s, 1H), 8.80 (s, 1H) ), 8.01–7.91 (m, 4H), 7.77 (d, J = 8.6 Hz, 1H), 7.69 (d, J = 7.5 Hz, 1H), 7.66–7.59 (m, 3H), 7.37 (d, J = 8.9Hz, 1H), 7.31 (d, J = 1.9 Hz, 1H), 7.24 (d, J = 8.6 Hz, 2H), 7.16 (d, J = 8.6 Hz, 2H), 7.05 (d, J = 1.4 Hz , 1H), 4.65 (dd, J = 14.0, 8.2 Hz, 1H), 3.12-3.02 (m, 2H), 2.53 (d, J = 1.9 Hz, 1H).
实施例55Example 55
合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((S)-2,3-二羟基丙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-((S)-2,3-Dihydroxypropoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000332
Figure PCTCN2020133493-appb-000332
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-(2,2-二甲基-1,3-二氧戊环-4-基)4-甲基苯磺酸甲酯Step A: Synthesis of (S)-(2,2-Dimethyl-1,3-dioxolane-4-yl)4-methylbenzenesulfonate methyl ester
Figure PCTCN2020133493-appb-000333
Figure PCTCN2020133493-appb-000333
将(R)-(2,2-二甲基-1,3-二氧戊环-4-基)甲醇(1.32克,10.0毫摩尔),4-二甲氨基吡啶(122毫克,1.0毫摩尔)和N,N-二异丙基乙胺(4.5毫升,30毫摩尔)溶于二氯甲烷(20.0毫升)中,冰水浴中搅拌5分钟,向反应液中加对甲基苯磺酰氯(1.9克,10.0毫摩尔),然后室温搅拌3小时。(R)-(2,2-Dimethyl-1,3-dioxolane-4-yl)methanol (1.32 g, 10.0 mmol), 4-dimethylaminopyridine (122 mg, 1.0 mmol) ) And N,N-diisopropylethylamine (4.5 mL, 30 mmol) were dissolved in dichloromethane (20.0 mL), stirred in an ice-water bath for 5 minutes, and p-toluenesulfonyl chloride ( 1.9 g, 10.0 mmol), and then stirred at room temperature for 3 hours.
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得到1.0克无色油状物(S)-(2,2-二甲基-1,3-二氧戊环-4-基)4-甲基苯磺酸甲酯(收率：35.0％)。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 to obtain 1.0 g of colorless oily (S)-(2,2-dimethyl-1,3-dioxolane-4-yl)4-methylbenzenesulfonic acid methane Esters (yield: 35.0%).
步骤B：合成(R)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶-1-羧酸叔丁酯Step B: Synthesis of (R)-4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000334
Figure PCTCN2020133493-appb-000334
将4-羟基哌啶-1-羧酸叔丁酯(460毫克,3.5毫摩尔)溶于N,N-二甲基甲酰胺(10.0毫升)中,冰水浴中搅拌5分钟,向反应液中加钠氢(183毫克,4.6毫摩尔),然后室温搅拌1小时。随后,向上述溶液中加入(S)-(2,2-二甲基-1,3-二氧戊环-4-基)4-甲基苯磺酸甲酯(1.0克,3.5毫摩尔),在50℃下搅拌5小时。Dissolve 4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (460 mg, 3.5 mmol) in N,N-dimethylformamide (10.0 ml), stir in an ice-water bath for 5 minutes, and add to the reaction solution Add sodium hydrogen (183 mg, 4.6 mmol), and then stir at room temperature for 1 hour. Subsequently, (S)-(2,2-dimethyl-1,3-dioxolane-4-yl)methyl 4-methylbenzenesulfonate (1.0 g, 3.5 mmol) was added to the above solution , Stir at 50°C for 5 hours.
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得到620毫克无色油状物(R)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶-1-羧酸叔丁酯(收率：56％)。The reaction was quenched by adding water to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). The organic phases were combined, and the organic phase was first saturated with brine (10 ml × 3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 620 mg of colorless oil (R )-4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxylic acid tert-butyl ester (Yield: 56%).
步骤C：合成(R)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶盐酸盐Step C: Synthesis of (R)-4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine hydrochloride
Figure PCTCN2020133493-appb-000335
Figure PCTCN2020133493-appb-000335
将(R)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶-1-羧酸叔丁酯(620毫克,2.0毫摩尔)溶于乙酸乙酯(6.0毫升)中。随后,向上述溶液中加入2.0M盐酸的乙酸乙酯溶液(2.0毫升),在室温下搅拌2小时。(R)-4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxylic acid tert-butyl ester (620 mg, 2.0 (Mmol) was dissolved in ethyl acetate (6.0 mL). Then, 2.0 M hydrochloric acid in ethyl acetate (2.0 mL) was added to the above solution, and the mixture was stirred at room temperature for 2 hours.
将反应液减压浓缩,得到400毫克白色固体(R)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶盐酸盐(收率：80％)。The reaction solution was concentrated under reduced pressure to obtain 400 mg of white solid (R)-4-(((2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine hydrochloride Salt (Yield: 80%).
步骤D：合成5-((S)-2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-(4-(((R)-2,2-二甲基-1,3-二氧戊环-4-基)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 D: Synthesis of 5-((S)-2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(((R)-2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxamido)phenyl) Propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000336
Figure PCTCN2020133493-appb-000336
室温下,将二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(200毫克,0.23毫摩尔)和(R)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶盐酸盐(170毫克,0.7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(0.5毫升,2.6毫摩尔)。在60℃反应12小时。At room temperature, di-tert-butyl(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylate (200 mg, 0.23 mmol) and (R) -4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine hydrochloride (170 mg, 0.7 mmol) dissolved in tetrahydrofuran (6.0 ml ). Then, add N,N-diisopropylethylamine (0.5 mL, 2.6 mmol) to the above solution and 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,所得残余物用硅胶柱层析纯化得到160毫克黄色固体5-((S)-2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-(4-(((R)-2,2-二甲基-1,3-二氧戊环-4-基)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：71％)。LCMS：RT＝4.53min,[M-H] -＝983.23。 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 The resulting residue was purified by silica gel column chromatography to obtain 160 mg of yellow solid 5-((S)-2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino )-2-oxoacetamido)-3-(4-(4-(((R)-2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)piper (Pyridine-1-carboxamido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (Yield: 71%). LCMS: RT=4.53min, [MH] - = 983.23.
步骤E：合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((S)-2,3-二羟基丙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step E: Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-((S)-2,3-Dihydroxypropoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000337
Figure PCTCN2020133493-appb-000337
将5-((S)-2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-(4-(((R)-2,2-二甲基-1,3-二氧戊环-4-基)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(150毫克,0.13毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。Add 5-((S)-2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(((R)-2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxamido)phenyl)propionamido )-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.13 mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 mL) was added to the above solution ), 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21毫克白色固体5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((S)-2,3-二羟基丙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：19％)。LCMS：RT＝3.25min,[M-H] -＝787.07。 1H NMR(400MHz,DMSO)δ13.03–12.76(m,1H),11.73(s,1H),10.73(s,1H),10.05(s,1H),9.84(s,1H),8.77(d,J＝8.3Hz,1H),8.43(s,1H),7.99(d,J＝2.3Hz,1H),7.95(s,1H),7.77(d,J＝8.6Hz,1H),7.62(dd,J＝8.6,2.4Hz,1H),7.43–7.28(m,2H),7.18–7.04(m,2H),5.76(s,1H),4.79–4.41(m,2H),3.76(d,J＝13.7Hz,2H),3.48(ddd,J＝14.7,10.4,5.1Hz,3H),3.10(dd,J＝15.8,7.2Hz,2H),2.55(s,2H),1.80(s,1H),1.38(d,J＝9.0Hz,1H)。 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 21 mg of white solid 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(4-((S)-2,3-dihydroxypropoxy)piperidine-1-carboxamido)phenylpropionamido ) -1H-indole-2-carboxylic acid (yield: 19%). LCMS: RT = 3.25 min, [MH] - =787.07. 1 H NMR (400MHz, DMSO) δ 13.03-12.76 (m, 1H ), 11.73 (s, 1H), 10.73 (s, 1H), 10.05 (s, 1H), 9.84 (s, 1H), 8.77 (d, J = 8.3 Hz, 1H), 8.43 (s, 1H), 7.99 (d,J=2.3Hz,1H),7.95(s,1H),7.77(d,J=8.6Hz,1H), 7.62(dd,J=8.6,2.4Hz,1H),7.43-7.28(m, 2H), 7.18–7.04 (m, 2H), 5.76 (s, 1H), 4.79–4.41 (m, 2H), 3.76 (d, J = 13.7 Hz, 2H), 3.48 (ddd, J = 14.7, 10.4, 5.1 Hz, 3H), 3.10 (dd, J=15.8, 7.2 Hz, 2H), 2.55 (s, 2H), 1.80 (s, 1H), 1.38 (d, J=9.0 Hz, 1H).
实施例56Example 56
合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((R)-2,3-二羟基丙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-((R)-2,3-Dihydroxypropoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000338
Figure PCTCN2020133493-appb-000338
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(R)-(2,2-二甲基-1,3-二氧戊环-4-基)4-甲基苯磺酸甲酯Step A: Synthesis of (R)-(2,2-Dimethyl-1,3-dioxolane-4-yl)4-methylbenzenesulfonate methyl ester
Figure PCTCN2020133493-appb-000339
Figure PCTCN2020133493-appb-000339
将(S)-(2,2-二甲基-1,3-二氧戊环-4-基)甲醇(1.32克,10.0毫摩尔),4-二甲氨基吡啶(122毫克,1.0毫摩尔)和N,N-二异丙基乙胺(4.5毫升,30毫摩尔)溶于二氯甲烷(20.0毫升)中,冰水浴中搅拌5分钟,向反应液中加对甲基苯磺酰氯(1.9克,10.0毫摩尔),然后室温搅拌3小时。(S)-(2,2-Dimethyl-1,3-dioxolane-4-yl)methanol (1.32 g, 10.0 mmol), 4-dimethylaminopyridine (122 mg, 1.0 mmol) ) And N,N-diisopropylethylamine (4.5 mL, 30 mmol) were dissolved in dichloromethane (20.0 mL), stirred in an ice-water bath for 5 minutes, and p-toluenesulfonyl chloride ( 1.9 g, 10.0 mmol), and then stirred at room temperature for 3 hours.
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得到1.0克无色油状物(R)-(2,2-二甲基-1,3-二氧戊环-4-基)4-甲基苯磺酸甲酯(收率：35.0％)。The reaction was quenched by adding water to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). The organic phases were combined, and the organic phase was first saturated with brine (10 ml × 3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 1.0 g of colorless oil (R )-(2,2-Dimethyl-1,3-dioxolane-4-yl)methyl 4-methylbenzenesulfonate (yield: 35.0%).
步骤B：合成(S)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶-1-羧酸叔丁酯Step B: Synthesis of (S)-4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000340
Figure PCTCN2020133493-appb-000340
将4-羟基哌啶-1-羧酸叔丁酯(460毫克,3.5毫摩尔)溶于N,N-二甲基甲酰胺(10.0毫升)中,冰水浴中搅拌5分钟,向反应液中加钠氢(183毫克,4.6毫摩尔),然后室温搅拌1小时。随后,向上述溶液中加入(R)-(2,2-二甲基-1,3-二氧戊环-4-基)4-甲基苯磺酸甲酯(1.0克,3.5毫摩尔),在50℃下搅拌5小时。Dissolve 4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (460 mg, 3.5 mmol) in N,N-dimethylformamide (10.0 ml), stir in an ice-water bath for 5 minutes, and add to the reaction solution Add sodium hydrogen (183 mg, 4.6 mmol), and then stir at room temperature for 1 hour. Subsequently, (R)-(2,2-dimethyl-1,3-dioxolane-4-yl)methyl 4-methylbenzenesulfonate (1.0 g, 3.5 mmol) was added to the above solution , Stir at 50°C for 5 hours.
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得到620毫克无色油状物(S)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶-1-羧酸叔丁酯(收率：56％)。The reaction was quenched by adding water to the reaction solution. The mixture was extracted with ethyl acetate (20 ml×3 times). The organic phases were combined, and the organic phase was first saturated with brine (10 ml × 3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 620 mg of colorless oil (S )-4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxylic acid tert-butyl ester (Yield: 56%).
步骤C：合成(S)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶盐酸盐Step C: Synthesis of (S)-4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine hydrochloride
Figure PCTCN2020133493-appb-000341
Figure PCTCN2020133493-appb-000341
将(S)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶-1-羧酸叔丁酯(620毫克,2.0毫摩尔)溶于乙酸乙酯(6.0毫升)中。随后,向上述溶液中加入2.0M盐酸的乙酸乙酯溶液(2.0毫升),在室温下搅拌2小时。(S)-4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxylic acid tert-butyl ester (620 mg, 2.0 (Mmol) was dissolved in ethyl acetate (6.0 mL). Then, 2.0 M hydrochloric acid in ethyl acetate (2.0 mL) was added to the above solution, and the mixture was stirred at room temperature for 2 hours.
将反应液减压浓缩,得到400毫克白色固体(S)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶盐酸盐(收率：80％)。The reaction solution was concentrated under reduced pressure to obtain 400 mg of white solid (S)-4-(((2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine hydrochloride Salt (Yield: 80%).
步骤D：合成5-((S)-2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-(4-(((S)-2,2-二甲基-1,3-二氧戊环-4-基)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 D: Synthesis of 5-((S)-2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(((S)-2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxamido)phenyl) Propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000342
Figure PCTCN2020133493-appb-000342
室温下,将二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(200毫克,0.23毫摩尔)和(S)-4-(((2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶盐酸盐(170毫克,0.7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(0.5毫升,2.6毫摩尔)。在60℃反应12小时。At room temperature, di-tert-butyl(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylate (200 mg, 0.23 mmol) and (S) -4-(((2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine hydrochloride (170 mg, 0.7 mmol) dissolved in tetrahydrofuran (6.0 ml ). Then, add N,N-diisopropylethylamine (0.5 mL, 2.6 mmol) to the above solution and 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160毫克黄色固体5-((S)-2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)二叔丁基)-3-(4-(4-(((S)-2,2-二甲基-1,3-二氧戊环-4-基)甲氧基)哌啶-1-甲酰胺基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(收率： 71％)。LCMS：RT＝4.53min,[M-H] -＝983.23。 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 160 mg of yellow solid 5-((S)-2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) two Tert-butyl)-3-(4-(4-(((S)-2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-methyl Amido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid ester (yield: 71%). LCMS: RT=4.53 min, [MH] - =983.23.
步骤E：合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((R)-2,3-二羟基丙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step E: Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-((R)-2,3-Dihydroxypropoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000343
Figure PCTCN2020133493-appb-000343
将5-((S)-2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)-3-(4-(4-(((S)-2,2-二甲基-1,3-二氧戊环-4-基)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(150毫克,0.13毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。Add 5-((S)-2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(((S)-2,2-Dimethyl-1,3-dioxolane-4-yl)methoxy)piperidine-1-carboxamido)phenyl)propionamido )-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.13 mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 mL) was added to the above solution ), 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21毫克白色固体5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((R)-2,3-二羟基丙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：19％)。LCMS：RT＝3.25min,[M-H] -＝787.07。 1H NMR(400MHz,DMSO)δ13.03–12.76(m,1H),11.73(s,1H),10.73(s,1H),10.05(s,1H),9.84(s,1H),8.77(d,J＝8.3Hz,1H),8.43(s,1H),7.99(d,J＝2.3Hz,1H),7.95(s,1H),7.77(d,J＝8.6Hz,1H),7.62(dd,J＝8.6,2.4Hz,1H),7.43–7.28(m,2H),7.18–7.04(m,2H),5.76(s,1H),4.79–4.41(m,2H),3.76(d,J＝13.7Hz,2H),3.48(ddd,J＝14.7,10.4,5.1Hz,3H),3.10(dd,J＝15.8,7.2Hz,2H),2.55(s,2H),1.80(s,1H),1.38(d,J＝9.0Hz,1H)。 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 21 mg of white solid 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(4-((R)-2,3-dihydroxypropoxy)piperidine-1-carboxamido)phenylpropionamido ) -1H-indole-2-carboxylic acid (yield: 19%). LCMS: RT = 3.25 min, [MH] - =787.07. 1 H NMR (400MHz, DMSO) δ 13.03-12.76 (m, 1H ), 11.73 (s, 1H), 10.73 (s, 1H), 10.05 (s, 1H), 9.84 (s, 1H), 8.77 (d, J = 8.3 Hz, 1H), 8.43 (s, 1H), 7.99 (d,J=2.3Hz,1H),7.95(s,1H),7.77(d,J=8.6Hz,1H), 7.62(dd,J=8.6,2.4Hz,1H),7.43-7.28(m, 2H), 7.18–7.04 (m, 2H), 5.76 (s, 1H), 4.79–4.41 (m, 2H), 3.76 (d, J = 13.7 Hz, 2H), 3.48 (ddd, J = 14.7, 10.4, 5.1 Hz, 3H), 3.10 (dd, J=15.8, 7.2 Hz, 2H), 2.55 (s, 2H), 1.80 (s, 1H), 1.38 (d, J=9.0 Hz, 1H).
实施例57Example 57
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000344
Figure PCTCN2020133493-appb-000344
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-((2-羟乙基)(甲基)氨基甲酰基)哌啶-1-甲酸叔丁酯Step A: Synthesis of tert-butyl 4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000345
Figure PCTCN2020133493-appb-000345
将1-(叔丁氧羰基)哌啶-4-羧酸(902毫克,3.9毫摩尔)和和N,N-二异丙基乙胺(1.9毫升,11.8毫摩尔)溶于1,2-二氯乙烷(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(2.2克,5.9毫摩尔)和2-(甲基氨基)乙-1-醇(585毫克,7.8毫摩尔)。在室温下搅拌2小时。Dissolve 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (902 mg, 3.9 mmol) and N,N-diisopropylethylamine (1.9 mL, 11.8 mmol) in 1,2- Dichloroethane (3.0 mL). Subsequently, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.2 g, 5.9 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.2 g, 5.9 mmol) were added to the above solution. (Methylamino)-1-ol (585 mg, 7.8 mmol). Stir at room temperature for 2 hours.
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得到1.0克黄色固体4-((2-羟乙基)(甲基)氨基甲酰基)哌啶-1-甲酸叔丁酯(收率：100％)。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 to obtain 1.0 g of tert-butyl 4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxylate as a yellow solid (yield: 100%).
步骤B：合成N-(2-羟乙基)-N-甲基哌啶-4-羧酰胺的三氟乙酸盐Step B: Synthesis of the trifluoroacetate salt of N-(2-hydroxyethyl)-N-methylpiperidine-4-carboxamide
Figure PCTCN2020133493-appb-000346
Figure PCTCN2020133493-appb-000346
将4-((2-羟乙基)(甲基)氨基甲酰基)哌啶-1-甲酸叔丁酯(1.0克,3.9毫摩尔)溶于二氯甲烷(6.0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-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxylate (1.0 g, 3.9 mmol) was dissolved in dichloromethane (6.0 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N-(2-羟乙基)-N-甲基哌啶-4-羧酰胺的三氟乙酸盐(收率：100％)。The reaction solution was concentrated under reduced pressure in an air bath to obtain 1.0 g of trifluoroacetate of yellow solid N-(2-hydroxyethyl)-N-methylpiperidine-4-carboxamide (yield: 100%).
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((2-羟乙基)(甲基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-di Di-tert-butyl carboxylate
Figure PCTCN2020133493-appb-000347
Figure PCTCN2020133493-appb-000347
室温下,将二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(120毫克,0.14毫摩尔)和N-(2-羟乙基)-N-甲基哌啶-4-羧酰胺的三氟乙酸盐(160毫克,0.56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,1.7毫摩尔)。在60℃反应12小时。At room temperature, di-tert-butyl(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylate (120 mg, 0.14 mmol) and N-( The trifluoroacetate (160 mg, 0.56 mmol) of 2-hydroxyethyl)-N-methylpiperidine-4-carboxamide was dissolved in tetrahydrofuran (6.0 mL). Then, N, N -Diisopropylethylamine (0.3 mL, 1.7 mmol). 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,所得残余物用硅胶柱层析纯化得到78毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((2-羟乙基)(甲基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：57％)。LCMS：RT＝4.16min,[M-H] -＝954.33。 The reaction solution was quenched by adding water, the mixed solution was extracted with ethyl acetate (30 ml × 3 times), the organic phase was combined, and the organic phase was saturated brine (20 ml × 3 times), dried with anhydrous sodium sulfate, and concentrated under reduced pressure The resulting residue was purified by silica gel column chromatography to obtain 78 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino )-2-oxoacetamido)-3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido) -1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 57%). LCMS: RT = 4.16 min, [MH] - =954.33.
步骤D：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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 D: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000348
Figure PCTCN2020133493-appb-000348
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((2-羟乙基)(甲基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(78毫克,0.08毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester (78 mg, 0.08 mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 mL) was added to the above solution, and the mixture was 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14毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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：22％)。LCMS：RT＝3.26min,[M+H] +＝799.98。 1H NMR(400MHz,DMSO)δ12.91(s,1H),11.72(s,1H),10.72(s,1H),10.04(s,1H),9.84(s,1H),8.76(d,J＝8.4Hz,1H),8.40(s,1H),8.00(d,J＝2.2Hz,1H),7.94(s,1H),7.78(dd,J＝8.6,2.1Hz,1H),7.62(dt,J＝8.6,2.5Hz,1H),7.34(ddd,J＝10.8,10.1,5.4Hz,4H),7.12(d,J＝8.6Hz,2H),7.06(d,J＝2.1Hz,1H),4.72–4.59(m,2H),4.20–4.00(m,2H),3.59–3.39(m,3H),3.08(s,3H),2.95–2.83(m,2H),2.81(s,2H),2.77(s,1H),2.61(s,1H),2.06–1.85(m,2H),1.62(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 14 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(4-((2-hydroxyethyl)(methyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamide Group) -1H-indole-2-carboxylic acid (yield: 22%). LCMS: RT = 3.26 min, [M+H] + = 799.98. 1 H NMR (400MHz, DMSO) δ 12.91 (s, 1H), 11.72 (s, 1H), 10.72 (s, 1H), 10.04 (s, 1H), 9.84 (s, 1H), 8.76 (d, J = 8.4 Hz, 1H), 8.40 (s, 1H), 8.00 (d, J = 2.2 Hz, 1H), 7.94 (s, 1H), 7.78 (dd, J = 8.6, 2.1 Hz, 1H), 7.62 (dt, J = 8.6, 2.5 Hz, 1H), 7.34 (ddd ,J = 10.8, 10.1, 5.4 Hz, 4H), 7.12 (d, J = 8.6 Hz, 2H), 7.06 (d, J = 2.1 Hz, 1H), 4.72-4.59 (m, 2H), 4.20-4.00 ( m, 2H), 3.59–3.39(m, 3H), 3.08(s, 3H), 2.95–2.83(m, 2H), 2.81(s, 2H), 2.77(s, 1H), 2.61(s, 1H) ,2.06–1.85(m,2H),1.62(s,2H).
实施例58Example 58
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000349
Figure PCTCN2020133493-appb-000349
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-((2-羟乙基)氨基甲酰基)哌啶-1-甲酸叔丁酯Step A: Synthesis of tert-butyl 4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000350
Figure PCTCN2020133493-appb-000350
将1-(叔丁氧羰基)哌啶-4-羧酸(902毫克,3.9毫摩尔)和和N,N-二异丙基乙胺(1.9毫升,11.8毫摩尔)溶于1,2-二氯乙烷(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(2.2克,5.9毫摩尔)和2-氨基乙醇(476毫克,7.8毫摩尔)。在室温下搅拌2小时。Dissolve 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (902 mg, 3.9 mmol) and N,N-diisopropylethylamine (1.9 mL, 11.8 mmol) in 1,2- Dichloroethane (3.0 mL). Subsequently, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.2 g, 5.9 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.2 g, 5.9 mmol) were added to the above solution. Aminoethanol (476 mg, 7.8 mmol). Stir at room temperature for 2 hours.
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。所得残余物直接用于下一步。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 resulting residue was used directly in the next step.
步骤B：合成N-(2-羟乙基)-哌啶-4-羧酰胺的三氟乙酸盐Step B: Synthesis of the trifluoroacetate salt of N-(2-hydroxyethyl)-piperidine-4-carboxamide
Figure PCTCN2020133493-appb-000351
Figure PCTCN2020133493-appb-000351
将4-((2-羟乙基)氨基甲酰基)哌啶-1-甲酸叔丁酯(1.0克,3.9毫摩尔)溶于二氯甲烷(6.0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-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxylate (1.0 g, 3.9 mmol) was dissolved in dichloromethane (6.0 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0.8克黄色固体N-(2-羟乙基)哌啶-4-羧酰胺的三氟乙酸盐(收率：100％)。The reaction solution was concentrated under reduced pressure in an air bath to obtain 0.8 g of trifluoroacetic acid salt of N-(2-hydroxyethyl)piperidine-4-carboxamide as a yellow solid (yield: 100%).
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((2-羟乙基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid Butyl
Figure PCTCN2020133493-appb-000352
Figure PCTCN2020133493-appb-000352
室温下,将二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(180毫克,0.21毫摩尔)和N-(2-羟乙基)哌啶-4-羧酰胺的三氟乙酸盐(160毫克,0.56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,1.7毫摩尔)。在60℃反应12小时。At room temperature, di-tert-butyl(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylate (180 mg, 0.21 mmol) and N-( The trifluoroacetate (160 mg, 0.56 mmol) of 2-hydroxyethyl)piperidine-4-carboxamide was dissolved in tetrahydrofuran (6.0 mL). Then, N,N-diisopropyl was added to the above solution. Ethylamine (0.3 mL, 1.7 mmol). Reacted 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,所得残余物用硅胶柱层析纯化得到150毫克黄色固体二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((2-羟乙基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸酯(收率：76％)。LCMS：RT＝4.08min,[M-H] -＝942.23。 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 The resulting residue was purified by silica gel column chromatography to obtain 150 mg of yellow solid di-tert-butyl (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl) (Phenyl)amino)-2-oxoacetamido)-3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido) -1H-indole-1,2-dicarboxylate (yield: 76%). LCMS: RT = 4.08 min, [MH] - =942.23.
步骤D：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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 D: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000353
Figure PCTCN2020133493-appb-000353
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((2-羟乙基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克,0.14毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester ( 150 mg, 0.14 mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 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22毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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：17％)。LCMS：RT＝3.22min,[M-H] -＝784.20。LCMS：RT＝3.22min,[M-H] -＝784.20。 1H NMR(400MHz,DMSO)δ12.93(s,1H),11.73(s,1H),10.73(s,1H),10.05(s,1H),9.84(s,1H),8.78(d,J＝8.3Hz,1H),8.41(s,1H),8.11–7.90(m,1H),7.79(dd,J＝11.1,7.1Hz,1H),7.61(dd,J＝8.6,2.4Hz,1H),7.44–7.26(m,2H),7.16–7.02(m,2H),4.67(dd,J＝14.5,7.8Hz,1H),4.10(d,J＝13.1Hz,2H),3.10(dt,J＝11.4,5.8Hz,2H),2.83–2.65(m,1H),2.55(s,1H),2.53–2.52(m,1H),2.40–2.26(m,1H),1.67(d,J＝10.4Hz,1H),1.47(dd,J＝21.5,12.1Hz,1H)。 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 22 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(4-((2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H -Indole-2-carboxylic acid (yield: 17%). LCMS: RT = 3.22 min, [MH] - = 784.20. LCMS: RT = 3.22 min, [MH] - = 784.20. 1 H NMR (400MHz, DMSO)δ 12.93(s, 1H), 11.73(s, 1H), 10.73(s, 1H), 10.05(s, 1H), 9.84(s, 1H), 8.78(d, J=8.3Hz, 1H) ,8.41(s,1H),8.11–7.90(m,1H),7.79(dd,J＝11.1,7.1Hz,1H), 7.61(dd,J＝8.6,2.4Hz,1H),7.44-7.26(m ,2H),7.16–7.02(m,2H),4.67(dd,J=14.5,7.8Hz,1H),4.10(d,J=13.1Hz,2H),3.10(dt,J=11.4,5.8Hz, 2H), 2.83–2.65(m,1H), 2.55(s,1H), 2.53–2.52(m,1H), 2.40–2.26(m,1H), 1.67(d,J=10.4Hz,1H), 1.47 (dd, J=21.5, 12.1 Hz, 1H).
实施例59Example 59
合成(S)-5-(3-(4-(4-(双(2-羟乙基)氨基甲酰基)哌啶-1-甲酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(3-(4-(4-(bis(2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)-2-(2-((5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000354
Figure PCTCN2020133493-appb-000354
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(双(2-羟乙基)氨基甲酰基)哌啶-1-甲酸叔丁酯Step A: Synthesis of tert-butyl 4-(bis(2-hydroxyethyl)carbamoyl)piperidine-1-carboxylate
Figure PCTCN2020133493-appb-000355
Figure PCTCN2020133493-appb-000355
将1-(叔丁氧羰基)哌啶-4-羧酸(902毫克,3.9毫摩尔)和和N,N-二异丙基乙胺(1.9毫升,11.8毫摩尔)溶于1,2-二氯乙烷(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(2.2克,5.9毫摩尔)和2,2'-氮杂二基双(乙烷-1-醇)(820毫克,7.8毫摩尔)。在室温下搅拌2小时。Dissolve 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (902 mg, 3.9 mmol) and N,N-diisopropylethylamine (1.9 mL, 11.8 mmol) in 1,2- Dichloroethane (3.0 mL). Subsequently, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.2 g, 5.9 mmol) and 2, 2'-Azadiylbis(ethane-1-ol) (820 mg, 7.8 mmol). Stir at room temperature for 2 hours.
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。所得残余物直接用于下一步。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 resulting residue was used directly in the next step.
步骤B：合成N,N-双(2-羟乙基)哌啶-4-羧酰胺的三氟乙酸盐Step B: Synthesis of the trifluoroacetate salt of N,N-bis(2-hydroxyethyl)piperidine-4-carboxamide
Figure PCTCN2020133493-appb-000356
Figure PCTCN2020133493-appb-000356
将4-(双(2-羟乙基)氨基甲酰基)哌啶-1-甲酸叔丁酯(1.0克,3.9毫摩尔)溶于二氯甲烷(6.0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-(bis(2-hydroxyethyl)carbamoyl)piperidine-1-carboxylate (1.0 g, 3.9 mmol) was dissolved in dichloromethane (6.0 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1.2克黄色固体N，N-双(2-羟乙基)哌啶-4-羧酰胺的三氟乙酸盐(收率：100％)。The reaction solution was concentrated under reduced pressure in an air bath to obtain 1.2 g of trifluoroacetate of yellow solid N,N-bis(2-hydroxyethyl)piperidine-4-carboxamide (yield: 100%).
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(双(2-羟乙基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(bis(2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid Tert-butyl ester
Figure PCTCN2020133493-appb-000357
Figure PCTCN2020133493-appb-000357
室温下,将二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(150毫克,0.17毫摩尔)和N，N-双(2-羟乙基)哌啶-4-羧酰胺的三氟乙酸盐(220毫克,0.70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,1.7毫摩尔)。在60℃反应12小时。At room temperature, di-tert-butyl(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylate (150 mg, 0.17 mmol) and N, N -Bis(2-hydroxyethyl)piperidine-4-carboxamide trifluoroacetate (220 mg, 0.70 mmol) was dissolved in tetrahydrofuran (6.0 ml). Then, N,N-di Isopropylethylamine (0.3 mL, 1.7 mmol). Reacted 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,所得残余物用硅胶柱层析纯化得到110毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(双(2-羟乙基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：26％)。LCMS：RT＝4.13min,[M+Na] +＝964.24。 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 The resulting residue was purified by silica gel column chromatography to obtain 110 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino )-2-oxoacetamido)-3-(4-(4-(bis(2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H- Indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 26%). LCMS: RT = 4.13 min, [M+Na] + =964.24.
步骤D：合成(S)-5-(3-(4-(4-(双(2-羟乙基)氨基甲酰基)哌啶-1-甲酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸Step D: Synthesis of (S)-5-(3-(4-(4-(bis(2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenyl)-2-(2- ((5-Chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetylamino)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000358
Figure PCTCN2020133493-appb-000358
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(双(2-羟乙基)氨基甲酰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克,0.14毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(bis(2-hydroxyethyl)carbamoyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.14 mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 mL) was added to the above solution, and the mixture was 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毫克白色固体(S)-5-(3-(4-(4-(双(2-羟乙基)氨基甲酰基)哌啶-1-甲酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸(收率：4％),LCMS：RT＝2.85min,[M-H] -＝828.20。以及3毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-((2-((2-羟乙基)氨基)乙氧基)羰基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：3％)。LCMS：RT＝3.21min,[M-H] -＝828.20。 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 white solid (S)-5-(3-(4-(4-(bis(2-hydroxyethyl)carbamoyl)piperidine-1 -Carboxamido)phenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H -Indole-2-carboxylic acid (yield: 4%), LCMS: RT = 2.85 min, [MH] - =828.20. And 3 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-((2-((2-hydroxyethyl)amino)ethoxy)carbonyl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2 -Carboxylic acid (yield: 3%). LCMS: RT = 3.21 min, [MH] - = 828.20.
实施例60Example 60
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-(Methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000359
Figure PCTCN2020133493-appb-000359
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(甲基磺酰基)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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000360
Figure PCTCN2020133493-appb-000360
室温下,将二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(170毫克,0.2毫摩尔)和甲磺酰胺(80毫克,0.8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,2.4毫摩尔)。在60摄氏度反应12小时。At room temperature, di-tert-butyl(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylate (170 mg, 0.2 mmol) and methanesulfonamide (80 mg, 0.8 mmol) was dissolved in tetrahydrofuran (6.0 mL). Then, N,N-diisopropylethylamine (0.3 mL, 2.4 mmol) was added to the above solution and reacted at 60 degrees Celsius 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,所得残余物用硅胶柱层析纯化得到220毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(甲基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：53％)。LCMS：RT＝4.32min,[M+H] +＝863.18。 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 The resulting residue was purified by silica gel column chromatography to obtain 220 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino )-2-oxoacetamido)-3-(4-(3-(methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid Butyl ester (yield: 53%). LCMS: RT = 4.32 min, [M+H] + =863.18.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-(Methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000361
Figure PCTCN2020133493-appb-000361
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(甲基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(220毫克,0.13毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(3-(methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (220 mg, 0.13 mmol) dissolved in dichloro Methane (2.0 mL). Then, trifluoroacetic acid (0.5 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13毫克黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(甲基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：14％)。LCMS：RT＝3.44min,[M+H] +＝709.07。 1H NMR(400MHz,DMSO)δ11.72(s,1H),10.73(s,1H),10.05(s,1H),9.89–9.77(m,1H),8.79(d,J＝8.3Hz,1H),8.71(s,1H),7.98(dd,J＝11.6,9.3Hz,2H),7.77(d,J＝8.6Hz,1H),7.62(dd,J＝8.6,2.4Hz,1H),7.34(ddd,J＝11.6,10.9,5.4Hz,4H),7.18(d,J＝8.5Hz,2H),7.08(t,J＝10.4Hz,1H),6.67(s,1H),5.33(t,J＝5.1Hz,1H),4.68(dd,J＝14.0,8.2Hz,1H),4.10(d,J＝4.9Hz,1H),3.24–3.14(m,3H),3.10(s,2H),2.55(d,J＝2.9Hz,2H),2.01(dd,J＝14.4,6.9Hz,2H),1.47(s,1H)。 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 13 mg of yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(3-(methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield : 14%). LCMS: RT = 3.44 min, [M+H] + =709.07. 1 H NMR(400MHz,DMSO)δ11.72(s,1H), 10.73(s,1H), 10.05(s,1H), 9.89–9.77(m,1H), 8.79(d,J=8.3Hz,1H ), 8.71 (s, 1H), 7.98 (dd, J = 11.6, 9.3 Hz, 2H), 7.77 (d, J = 8.6 Hz, 1H), 7.62 (dd, J = 8.6, 2.4 Hz, 1H), 7.34 (ddd, J = 11.6, 10.9, 5.4 Hz, 4H), 7.18 (d, J = 8.5 Hz, 2H), 7.08 (t, J = 10.4 Hz, 1H), 6.67 (s, 1H), 5.33 (t, J = 5.1 Hz, 1H), 4.68 (dd, J = 14.0, 8.2 Hz, 1H), 4.10 (d, J = 4.9 Hz, 1H), 3.24-3.14 (m, 3H), 3.10 (s, 2H), 2.55 (d, J=2.9 Hz, 2H), 2.01 (dd, J=14.4, 6.9 Hz, 2H), 1.47 (s, 1H).
实施例61Example 61
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000362
Figure PCTCN2020133493-appb-000362
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(乙基磺酰基)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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000363
Figure PCTCN2020133493-appb-000363
室温下,将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(170毫克,0.2毫摩尔)和乙磺酰胺(90毫克,0.8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,2.4毫摩尔)。在60摄氏度反应12小时。At room temperature, add (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (170 mg, 0.2 mmol) and ethanesulfonamide ( 90 mg, 0.8 mmol) was dissolved in tetrahydrofuran (6.0 mL). Then, N,N-diisopropylethylamine (0.3 mL, 2.4 mmol) was added to the above solution and reacted at 60 degrees Celsius 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,所得残余物用硅胶柱层析纯化得到86毫克黄色固体二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(乙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(收率：49％)。LCMS：RT＝4.67min,[M+H] +＝886.22。 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 The resulting residue was purified by silica gel column chromatography to obtain 86 mg of yellow solid di-tert-butyl (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl) (Phenyl)amino)-2-oxoacetamido)-3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-di Carboxylic acid ester (yield: 49%). LCMS: RT = 4.67 min, [M+H] + =886.22.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000364
Figure PCTCN2020133493-appb-000364
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(乙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(86毫克,0.1毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (86 mg, 0.1 mmol) dissolved in dichloride Methane (2.0 mL). Then, trifluoroacetic acid (0.5 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15毫克黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(乙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：21％)。LCMS：RT＝3.48min,[M+H] +＝721.21。 1H NMR(400MHz,DMSO)δ13.06–12.83(m,1H),11.73(s,1H),10.73(s,1H),10.06(s,1H),9.84(d,J＝5.1Hz,1H),8.87–8.76(m,2H),8.04–7.91(m,2H),7.77(d,J＝8.6Hz,1H),7.62(dd,J＝8.6,2.4Hz,1H),7.44–7.28(m,4H),7.21(d,J＝8.5Hz,2H),7.07(t,J＝5.7Hz,1H),5.76(s,1H),4.68(dd,J＝14.1,8.2Hz,1H),3.49–3.40(m,2H),3.12(s,1H),2.57–2.52(m,2H),1.24(dt,J＝11.3,7.4Hz,3H)。 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 15 mg of yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(3-(ethylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield :twenty one%). LCMS: RT = 3.48 min, [M+H] + =721.21. 1 H NMR (400MHz, DMSO) δ 13.06-12.83 (m, 1H), 11.73 (s, 1H), 10.73 (s, 1H), 10.06 (s, 1H), 9.84 (d, J = 5.1 Hz, 1H ), 8.87–8.76 (m, 2H), 8.04–7.91 (m, 2H), 7.77 (d, J = 8.6 Hz, 1H), 7.62 (dd, J = 8.6, 2.4 Hz, 1H), 7.44–7.28 ( m, 4H), 7.21 (d, J = 8.5 Hz, 2H), 7.07 (t, J = 5.7 Hz, 1H), 5.76 (s, 1H), 4.68 (dd, J = 14.1, 8.2 Hz, 1H), 3.49–3.40(m, 2H), 3.12(s, 1H), 2.57–2.52(m, 2H), 1.24(dt, J=11.3, 7.4Hz, 3H).
实施例62Example 62
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-(Methylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000365
Figure PCTCN2020133493-appb-000365
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基磺酰基)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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(cyclopropylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000366
Figure PCTCN2020133493-appb-000366
室温下,将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(170毫克,0.2毫摩尔)和环丙基磺酰胺(96毫克,0.8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,2.4毫摩尔)。在60摄氏度反应12小时。At room temperature, add (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (170 mg, 0.2 mmol) and cyclopropyl sulfonate The amide (96 mg, 0.8 mmol) was dissolved in tetrahydrofuran (6.0 mL). Then, N,N-diisopropylethylamine (0.3 mL, 2.4 mmol) was added to the above solution and reacted at 60 degrees Celsius 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,所得残余物用硅胶柱层析纯化得到50毫克黄色固体二叔丁基(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸酯(收率：28％)。LCMS：RT＝4.33min,[M+Na] +＝913.08。 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 The resulting residue was purified by silica gel column chromatography to obtain 50 mg of yellow solid di-tert-butyl (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl) (Phenyl)amino)-2-oxoacetamido)-3-(4-(3-(cyclopropylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2- Dicarboxylic acid ester (yield: 28%). LCMS: RT=4.33 min, [M+Na] + =913.08.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-(Cyclopropylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000367
Figure PCTCN2020133493-appb-000367
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(50毫克,0.06毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(3-(Cyclopropylsulfonyl)ureido)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (50 mg, 0.06 mmol) dissolved in two Chloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 mL) was added to the above solution, and the mixture was 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5毫克黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(环丙基磺酰基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：13％)。LCMS：RT＝3.52min,[M-H] -＝733.15。 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 5 mg of yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(3-(cyclopropylsulfonyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid (closed Rate: 13%). LCMS: RT = 3.52 min, [MH] - =733.15.
实施例63Example 63
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1,1-二氧化四氢-2H-硫代吡喃-4-基)-3-(2-羟乙基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)-3-(2-hydroxyethyl)ureido)phenyl)propionamido)-1H-indyl Dole-2-carboxylic acid
Figure PCTCN2020133493-appb-000368
Figure PCTCN2020133493-appb-000368
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(((2-((叔丁基二甲基甲硅烷基)氧基)乙基)氨基)四氢-2H-噻喃1,1-二氧化物Step A: Synthesis of 4-(((2-((tert-butyldimethylsilyl)oxy)ethyl)amino)tetrahydro-2H-thiopyran 1,1-dioxide
Figure PCTCN2020133493-appb-000369
Figure PCTCN2020133493-appb-000369
室温下,将4-氨基四氢-2H-噻喃1,1-二氧化物(500毫克,3.3毫摩尔)和碳酸铯(2.2克,6.6毫摩尔)溶于乙腈(20.0毫升)中。随后,向上述溶液中加(2-溴乙氧基)(叔丁基)二甲基硅烷(960毫克,4.0毫摩尔)。在80℃反应12小时。At room temperature, 4-aminotetrahydro-2H-thiopyran 1,1-dioxide (500 mg, 3.3 mmol) and cesium carbonate (2.2 g, 6.6 mmol) were dissolved in acetonitrile (20.0 mL). Subsequently, (2-bromoethoxy)(tert-butyl)dimethylsilane (960 mg, 4.0 mmol) was added to the above solution. React at 80°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,所得残余物用硅胶柱层析纯化得到600毫克黄色液体4-(((2-((叔丁基二甲基甲硅烷基)氧基)乙基)氨基)四氢-2H-噻喃1,1-二氧化物(收率：60％)。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 The resulting residue was purified by silica gel column chromatography to obtain 600 mg of yellow liquid 4-(((2-((tert-butyldimethylsilyl)oxy)ethyl)amino)tetrahydro-2H-thiopyran 1 ,1-Dioxide (yield: 60%).
步骤B：合成(S)-5-(3-(4-(3-(2-((叔丁基二甲基甲硅烷基)氧基)乙基)-3-(1,1-二氧化四氢-2H-噻喃-4-基)脲基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step B: Synthesis of (S)-5-(3-(4-(3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-3-(1,1-dioxide Tetrahydro-2H-thiopyran-4-yl)ureido)phenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2- Oxoacetamido) propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000370
Figure PCTCN2020133493-appb-000370
室温下,将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(150毫克,0.17毫摩尔)和4-(((2-((叔丁基二甲基甲硅烷基)氧基)乙基)氨基)四氢-2H-噻喃1,1-二氧化物(210毫克,0.68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,2.4毫摩尔)。在60℃反应12小时。At room temperature, add (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (150 mg, 0.17 mmol) and 4-(( (2-((tert-butyldimethylsilyl)oxy)ethyl)amino)tetrahydro-2H-thiopyran 1,1-dioxide (210 mg, 0.68 mmol) dissolved in tetrahydrofuran (6.0 ML). Then, add N,N-diisopropylethylamine (0.3 mL, 2.4 mmol) to the above solution and 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94毫克黄色固体(S)-5-(3-(4-(3-(2-((叔丁基二甲基甲硅烷基)氧基)乙基)-3-(1,1-二氧化四氢-2H-噻喃-4-基)脲基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：51％)。LCMS：RT＝4.98min,[M+H] +＝1077.33。 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 94 mg of yellow solid (S)-5-(3-(4-(3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-3-(1,1-di Tetrahydro-2H-thiopyran-4-yl)ureido)phenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 51%). LCMS: RT = 4.98 min, [M+H] + =1077.33.
步骤C：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1,1-二氧化四氢-2H-硫代吡喃-4-基)-3-(2-羟乙基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step C: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(1,1-tetrahydrodioxide-2H-thiopyran-4-yl)-3-(2-hydroxyethyl)ureido)phenyl)propionamido)- 1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000371
Figure PCTCN2020133493-appb-000371
将(S)-5-(3-(4-(3-(2-((叔丁基二甲基甲硅烷基)氧基)乙基)-3-(1,1-二氧化四氢-2H-噻喃-4-基)脲基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(94毫克,0.09毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。Add (S)-5-(3-(4-(3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-3-(1,1-tetrahydrodioxide- 2H-thiopyran-4-yl)ureido)phenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoethyl Amido)propionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (94 mg, 0.09 mmol) was dissolved in methylene chloride (2.0 mL). Subsequently, trifluoroacetic acid (0.5 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-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1,1-二氧化四氢-2H-硫代吡喃-4-基)-3-(2-羟乙基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：37％)。LCMS：RT＝3.36min,[M+H] +＝807.18。 1H NMR(500MHz,DMSO)δ12.92(s,1H),11.72(s,1H),10.72(s,1H),10.04(s,1H),9.83(s,1H),8.91(s,1H),8.76(d,J＝8.0Hz,1H),7.98(d,J＝2.3Hz,1H),7.94(s,1H),7.77(d,J＝8.6Hz,1H),7.61(dd,J＝8.6,2.3Hz,1H),7.37(d,J＝8.9Hz,1H),7.31(dd,J＝8.9,1.9Hz,1H),7.25(d,J＝8.5Hz,2H),7.13(d,J＝8.5Hz,2H),7.05(d,J＝1.5Hz,1H),5.32(t,J＝4.7Hz,1H),4.66(dd,J＝14.0,8.2Hz,1H),4.23(t,J＝12.2Hz,1H),3.58(t,J＝5.0Hz,2H),3.06(t,J＝10.6Hz,4H),2.54(s,2H),2.18(dd,J＝25.4,12.0Hz,2H),2.03–1.90(m,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 26 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)benzene) Yl)amino)-2-oxoacetamido)-3-(4-(3-(1,1-tetrahydro-2H-thiopyran-4-yl)-3-(2-hydroxy Ethyl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 37%). LCMS: RT = 3.36 min, [M+H] + =807.18. 1 H NMR (500MHz,DMSO)δ 12.92(s, 1H), 11.72(s, 1H), 10.72(s, 1H), 10.04(s, 1H), 9.83(s, 1H), 8.91(s, 1H), 8.76 (d,J=8.0Hz,1H),7.98(d,J=2.3Hz,1H),7.94(s,1H),7.77(d,J=8.6Hz,1H),7.61(dd,J=8.6, 2.3Hz, 1H), 7.37 (d, J = 8.9 Hz, 1H), 7.31 (dd, J = 8.9, 1.9 Hz, 1H), 7.25 (d, J = 8.5 Hz, 2H), 7.13 (d, J = 8.5Hz, 2H), 7.05 (d, J = 1.5 Hz, 1H), 5.32 (t, J = 4.7 Hz, 1H), 4.66 (dd, J = 14.0, 8.2 Hz, 1H), 4.23 (t, J = 12.2Hz, 1H), 3.58 (t, J = 5.0 Hz, 2H), 3.06 (t, J = 10.6 Hz, 4H), 2.54 (s, 2H), 2.18 (dd, J = 25.4, 12.0 Hz, 2H) ,2.03–1.90(m,4H).
实施例64Example 64
合成(S)-5-(2-(2-(((2-乙酰基5-氯苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-(((2-acetyl5-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)-1H-indole -2-carboxylic acid
Figure PCTCN2020133493-appb-000372
Figure PCTCN2020133493-appb-000372
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成1-(2-氨基-4-氯苯基)乙-1-酮Step A: Synthesis of 1-(2-amino-4-chlorophenyl)ethan-1-one
Figure PCTCN2020133493-appb-000373
Figure PCTCN2020133493-appb-000373
将1-(4-氯-2-硝基苯基)乙-1-酮(1.2克,6毫摩尔)溶于乙酸乙酯(50.0毫升)中。氮气保护后,冰水浴下加氯化亚锡二水合物(13.5克,60毫摩尔),缓慢升温到室温并搅拌12小时后。1-(4-Chloro-2-nitrophenyl)ethan-1-one (1.2 g, 6 mmol) was dissolved in ethyl acetate (50.0 mL). After nitrogen protection, stannous chloride dihydrate (13.5 g, 60 mmol) was added under ice-water bath, and the temperature was slowly raised to room temperature and stirred for 12 hours.
将反应液缓慢滴加到15％碳酸钠水溶液(150.0毫升)中,混合液用乙酸乙酯(60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得到0.7克黄色固体1-(2-氨基-4-氯苯基)乙-1-酮(收率：70％)。LCMS：RT＝3.85min,[M+H] +＝170.11。 The reaction solution was slowly added dropwise to 15% sodium carbonate aqueous solution (150.0 ml), the mixture was extracted with ethyl acetate (60 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (30 ml × 3 times) , Dry with anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 0.7 g of yellow solid 1-(2-amino-4-chlorophenyl)ethan-1-one (yield: 70%). LCMS: RT = 3.85 min, [M+H] + = 170.11.
步骤B：合成2-((2-乙酰基-5-氯苯基)氨基)-2-氧乙酸甲酯Step B: Synthesis of methyl 2-((2-acetyl-5-chlorophenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000374
Figure PCTCN2020133493-appb-000374
氮气保护下,将1-(2-氨基-4-氯苯基)乙-1-酮(700毫克,4.0毫摩尔)和吡啶(0.6毫升,9.0毫摩尔)溶于二氯甲烷(13.0毫升)中,冰水浴中搅拌5分钟,向反应液中缓慢滴加草酰氯单甲酯(612毫克,5.0毫摩尔)的二氯甲烷溶液(3.0毫升),冰水浴中搅拌1小时。Under nitrogen protection, dissolve 1-(2-amino-4-chlorophenyl)ethan-1-one (700 mg, 4.0 mmol) and pyridine (0.6 mL, 9.0 mmol) in dichloromethane (13.0 mL) After stirring in an ice-water bath for 5 minutes, slowly add dropwise a dichloromethane solution (3.0 ml) of monomethyl oxalyl chloride (612 mg, 5.0 mmol) to the reaction solution, and stir in an ice-water bath for 1 hour.
向反应液中加水(10.0毫升)淬灭反应,用乙酸乙酯(10毫升×3次)萃取。合并有机相,减压浓缩,所得残余物用硅胶柱层析纯化得到330毫克黄色固体2-((2-乙酰基-5-氯苯基)氨基)-2-氧乙酸甲酯(收率：31％)。LCMS：RT＝3.83min,[M+H] +＝256.00。 Water (10.0 mL) was added to the reaction solution to quench the reaction, and it was extracted with ethyl acetate (10 mL×3 times). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain 330 mg of yellow solid methyl 2-((2-acetyl-5-chlorophenyl)amino)-2-oxoacetate (yield: 31%). LCMS: RT = 3.83 min, [M+H] + =256.00.
步骤C：合成2-((2-乙酰基-5-氯苯基)氨基)-2-氧乙酸Step C: Synthesis of 2-((2-acetyl-5-chlorophenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000375
Figure PCTCN2020133493-appb-000375
将2-((2-乙酰基-5-氯苯基)氨基)-2-氧乙酸甲酯(330毫克,1.4毫摩尔)溶于四氢呋喃(18.0毫升)和水(6.0毫升)中。随后,向上述溶液中加入氢氧化锂一水合物(56毫克,1.4毫摩尔)。在室温下搅拌1小时。Methyl 2-((2-acetyl-5-chlorophenyl)amino)-2-oxoacetate (330 mg, 1.4 mmol) was dissolved in tetrahydrofuran (18.0 mL) and water (6.0 mL). Subsequently, lithium hydroxide monohydrate (56 mg, 1.4 mmol) was added to the above solution. Stir at room temperature for 1 hour.
加稀盐酸调节pH至弱酸性,减压浓缩除去大部分溶剂,加稀盐酸调节pH至3,析出大量固体,过滤,收集滤饼得到200毫克黄色固体2-((2-乙酰基-5-氯苯基)氨基)-2-氧乙酸(收率：63％)。LCMS：RT＝4.96min,[M-H] -＝239.92。 Add dilute hydrochloric acid to adjust the pH to weak acidity, concentrate under reduced pressure to remove most of the solvent, add dilute hydrochloric acid to adjust the pH to 3, precipitate a large amount of solids, filter, and collect the filter cake to obtain 200 mg of yellow solid 2-((2-acetyl-5- (Chlorophenyl)amino)-2-oxoacetic acid (Yield: 63%). LCMS: RT = 4.96 min, [MH] - =239.92.
步骤D：合成(S)-5-(2-(2-(2-((2-乙酰基-5-氯苯基)氨基)-2-氧乙酰氨基)-3-苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step D: Synthesis of (S)-5-(2-(2-(2-((2-acetyl-5-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) -1H-Indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000376
Figure PCTCN2020133493-appb-000376
将2-((2-乙酰基-5-氯苯基)氨基)-2-氧乙酸(120毫克,0.8毫摩尔),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(470毫克,1.2毫摩尔)和N,N-二异丙基乙胺(0.4毫升,2.4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(S)-5-(2-氨基-3-苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁基酯(470毫克,1.2毫摩尔),在室温下搅拌2小时。The 2-((2-acetyl-5-chlorophenyl)amino)-2-oxoacetic acid (120 mg, 0.8 mmol), 2-(7-azobenzotriazole)-N,N, N',N'-tetramethylurea hexafluorophosphate (470 mg, 1.2 mmol) and N,N-diisopropylethylamine (0.4 mL, 2.4 mmol) are dissolved in N,N-dimethyl Formamide (3.0 mL). Subsequently, (S)-5-(2-amino-3-phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (470 mg, 1.2 mmol) was added to the above solution. Mol), stirred at room temperature for 2 hours.
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得到55毫克黄色固体(S)-5-(2-(2-(2-((2-乙酰基-5-氯苯基)氨基)-2-氧乙酰氨基)-3-苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：31.0％)。LCMS：RT＝4.49min,[M+H] +＝603.15。 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 to obtain 55 mg of yellow solid (S)-5-(2-(2-(2-((2-acetyl-5-chlorophenyl)amino)-2-oxoacetamido )-3-Phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 31.0%). LCMS: RT = 4.49 min, [M+H] + =603.15 .
步骤E：合成(S)-5-(2-(2-(((2-乙酰基5-氯苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Step E: Synthesis of (S)-5-(2-(2-(((2-acetyl5-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)-1H -Indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000377
Figure PCTCN2020133493-appb-000377
将(S)-5-(2-(2-(2-((2-乙酰基-5-氯苯基)氨基)-2-氧乙酰氨基)-3-苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(55毫克,0.09毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(2-((2-Acetyl-5-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)-1H- Indole-1,2-dicarboxylic acid di-tert-butyl ester (55 mg, 0.09 mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 mL) was added to the above solution, and the mixture was heated at room temperature. Stir 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15毫克白色固体(S)-5-(2-(2-(((2-乙酰基5-氯苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：31％)。LCMS：RT＝4.03min,[M-H] -＝545.08。 1H NMR(500MHz,DMSO)δ12.80(s,1H),11.74(s,1H),10.07(s,1H),9.08(d,J＝8.5Hz,1H),8.67(dd,J＝10.6,2.2Hz,1H),8.15(d,J＝8.6Hz,2H),7.97(s,1H),7.41–7.36(m,2H),7.36–7.30(m,3H),7.28(t,J＝7.6Hz,2H),7.19(dd,J＝9.9,4.5Hz,1H),7.06(d,J＝1.4Hz,1H),5.32(t,J＝4.9Hz,1H),4.77(td,J＝8.7,5.3Hz,1H),3.22(d,J＝5.3Hz,1H),3.10(dt,J＝7.2,2.4Hz,1H),2.64(s,3H)。 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 15 mg of white solid (S)-5-(2-(2-(((2-acetyl5-chlorophenyl)amino)-2-oxyethane Amido)-3-phenylpropionamido)-1H-indole-2-carboxylic acid (yield: 31%). LCMS: RT = 4.03 min, [MH] - =545.08. 1 H NMR (500MHz, DMSO) δ 12.80 (s, 1H), 11.74 (s, 1H), 10.07 (s, 1H), 9.08 (d, J = 8.5 Hz, 1H), 8.67 (dd, J = 10.6, 2.2 Hz, 1H) ,8.15(d,J=8.6Hz,2H),7.97(s,1H),7.41-7.36(m,2H),7.36-7.30(m,3H),7.28(t,J=7.6Hz,2H), 7.19 (dd, J = 9.9, 4.5 Hz, 1H), 7.06 (d, J = 1.4 Hz, 1H), 5.32 (t, J = 4.9 Hz, 1H), 4.77 (td, J = 8.7, 5.3 Hz, 1H) ), 3.22 (d, J = 5.3 Hz, 1H), 3.10 (dt, J = 7.2, 2.4 Hz, 1H), 2.64 (s, 3H).
实施例65Example 65
合成(S)-5-(2-(2-(((2-(氨基甲基)-5-氯苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-(((2-(aminomethyl)-5-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)- 1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000378
Figure PCTCN2020133493-appb-000378
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成N-烯丙基-N-(4-氯-2-硝基苄基)丙-2-烯-1-胺Step A: Synthesis of N-allyl-N-(4-chloro-2-nitrobenzyl)prop-2-en-1-amine
Figure PCTCN2020133493-appb-000379
Figure PCTCN2020133493-appb-000379
将4-氯-2-硝基苯甲醛(2.2克,12毫摩尔)和醋酸(2.0毫升,36毫摩尔)溶于甲醇(40.0毫升)中。氮气保护后,加二烯丙基胺(2.3克,24毫摩尔),室温下搅拌2小时后,加氰基硼氢化钠(3.8克,60毫摩尔),4-Chloro-2-nitrobenzaldehyde (2.2 g, 12 mmol) and acetic acid (2.0 mL, 36 mmol) were dissolved in methanol (40.0 mL). After nitrogen protection, add diallylamine (2.3 g, 24 mmol), and after stirring for 2 hours at room temperature, add sodium cyanoborohydride (3.8 g, 60 mmol),
将反应液缓慢滴加到15％碳酸钠水溶液(150.0毫升)中,混合液用乙酸乙酯(60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得到1.5克无色油状物N-烯丙基-N-(4-氯-2-硝基苄基)丙-2-烯-1-胺(收率：46％)。LCMS：RT＝2.73min,[M+H] +＝267.09。 The reaction solution was slowly added dropwise to 15% sodium carbonate aqueous solution (150.0 ml), the mixture was extracted with ethyl acetate (60 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (30 ml × 3 times) , Dry with anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 1.5 g of colorless oily N-allyl-N-(4-chloro-2-nitrobenzyl)prop-2-en-1-amine (yield: 46 %). LCMS: RT = 2.73 min, [M+H] + =267.09.
步骤B：合成5-氯-2-((二烯丙基氨基)甲基)苯胺Step B: Synthesis of 5-chloro-2-((diallylamino)methyl)aniline
Figure PCTCN2020133493-appb-000380
Figure PCTCN2020133493-appb-000380
将N-烯丙基-N-(4-氯-2-硝基苄基)丙-2-烯-1-胺(1.1克,4毫摩尔)溶于乙酸乙酯(50.0毫升)中。氮气保护后,冰水浴下加氯化亚锡二水合物(9.2克,41毫摩尔),缓慢升温到室温并搅拌12小时后。N-allyl-N-(4-chloro-2-nitrobenzyl)prop-2-en-1-amine (1.1 g, 4 mmol) was dissolved in ethyl acetate (50.0 mL). After nitrogen protection, stannous chloride dihydrate (9.2 g, 41 mmol) was added under ice-water bath, slowly warming to room temperature and stirring for 12 hours.
将反应液缓慢滴加到15％碳酸钠水溶液(150.0毫升)中,混合液用乙酸乙酯(60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得到840毫克油状物5-氯-2-((二烯丙基氨基)甲基)苯胺(收率：65％)。LCMS：RT＝1.57min,[M+H] +＝237.10。 The reaction solution was slowly added dropwise to 15% sodium carbonate aqueous solution (150.0 ml), the mixture was extracted with ethyl acetate (60 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (30 ml × 3 times) , Dry with anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 840 mg of oily 5-chloro-2-((diallylamino)methyl)aniline (yield: 65%). LCMS: RT=1.57 min, [M+H] + =237.10.
步骤C：合成2-((5-氯-2-(((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-(((diallylamino)methyl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000381
Figure PCTCN2020133493-appb-000381
氮气保护下,将5-氯-2-((二烯丙基氨基)甲基)苯胺(840毫克,3.6毫摩尔)和吡啶(0.5毫升,7.1毫摩尔)溶于二氯甲烷(30.0毫升)中,冰水浴中搅拌5分钟,向反应液中缓慢滴加草酰氯单甲酯(0.4毫克,4.3毫摩尔)的二氯甲烷溶液(3.0毫升),冰水浴中搅拌1小时。Under nitrogen protection, dissolve 5-chloro-2-((diallylamino)methyl)aniline (840 mg, 3.6 mmol) and pyridine (0.5 mL, 7.1 mmol) in dichloromethane (30.0 mL) After stirring in an ice-water bath for 5 minutes, slowly add dropwise a dichloromethane solution (3.0 ml) of monomethyl oxalyl chloride (0.4 mg, 4.3 mmol) to the reaction solution, and stir in an ice-water bath for 1 hour.
向反应液中加水(10.0毫升)淬灭反应,用乙酸乙酯(10毫升×3次)萃取。合并有机相,减压浓缩,所得残余物用硅胶柱层析纯化得到700毫克黄色液体2-((5-氯-2-(((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酸甲酯(收率：31％)。LCMS：RT＝4.13min,[M+H] +＝323.11。 Water (10.0 mL) was added to the reaction solution to quench the reaction, and it was extracted with ethyl acetate (10 mL×3 times). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain 700 mg of yellow liquid 2-((5-chloro-2-(((diallylamino)methyl)phenyl)amino)- Methyl 2-oxoacetate (yield: 31%). LCMS: RT = 4.13 min, [M+H] + =323.11.
步骤D：合成2-((5-氯-2-(((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酸Step D: Synthesis of 2-((5-chloro-2-(((diallylamino)methyl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000382
Figure PCTCN2020133493-appb-000382
将2-((5-氯-2-(((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酸甲酯(700毫克,2.2毫摩尔)溶于四氢呋喃(18.0毫升)和水(6.0毫升)中。随后,向上述溶液中加入氢氧化锂一水合物(91毫克,2.2毫摩尔)。在室温下搅拌1小时。Methyl 2-((5-chloro-2-(((diallylamino)methyl)phenyl)amino)-2-oxoacetate (700 mg, 2.2 mmol) was dissolved in tetrahydrofuran (18.0 ml ) And water (6.0 mL). Then, lithium hydroxide monohydrate (91 mg, 2.2 mmol) was added to the above solution. Stir at room temperature for 1 hour.
加稀盐酸调节pH至弱酸性,减压浓缩除去大部分溶剂,加稀盐酸调节pH至3,析出大量固体,过滤,收集滤饼得到340毫克白色固体2-((5-氯-2-(((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酸(收率：50％)。LCMS：RT＝2.70min,[M-H] -＝309.23。 Add dilute hydrochloric acid to adjust the pH to weak acidity, concentrate under reduced pressure to remove most of the solvent, add dilute hydrochloric acid to adjust the pH to 3, precipitate a large amount of solids, filter, and collect the filter cake to obtain 340 mg of white solid 2-((5-chloro-2-( ((Diallylamino)methyl)phenyl)amino)-2-oxoacetic acid (yield: 50%). LCMS: RT = 2.70 min, [MH] - =309.23.
步骤D：合成(S)-5-(2-(2-((5-氯-2-((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚1,2-二羧酸二叔丁酯Step D: Synthesis of (S)-5-(2-(2-((5-chloro-2-((diallylamino)methyl)phenyl)amino)-2-oxoacetamido)- 3-Phenylpropionamido)-1H-indole 1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000383
Figure PCTCN2020133493-appb-000383
将2-((5-氯-2-(((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酸(160毫克,0.5毫摩尔),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(285毫克,0.75毫摩尔)和N,N-二异丙基乙胺(0.2毫升,1.2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(S)-5-(2-氨基-3-苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁基酯(228毫克,0.6毫摩尔),在室温下搅拌2小时。Add 2-((5-chloro-2-(((diallylamino)methyl)phenyl)amino)-2-oxoacetic acid (160 mg, 0.5 mmol), 2-(7-azo Benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (285mg, 0.75mmol) and N,N-diisopropylethylamine (0.2ml, 1.2ml) Mol) was dissolved in N,N-dimethylformamide (3.0 ml). Then, (S)-5-(2-amino-3-phenylpropionamido)-1H-indole was added to the above solution -Di-tert-butyl 1,2-dicarboxylic acid (228 mg, 0.6 mmol), stirred at room temperature for 2 hours.
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得到40毫克黄色固体(S)-5-(2-(2-((5-氯-2-((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚1,2-二羧酸二叔丁酯(收率：12％)。LCMS：RT＝4.49min,[M-H] -＝603.15。 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 to obtain 40 mg of yellow solid (S)-5-(2-(2-((5-chloro-2-((diallylamino)methyl)phenyl)amino) -2-oxoacetamido)-3-phenylpropionamido)-1H-indole 1,2-dicarboxylic acid di-tert-butyl ester (yield: 12%). LCMS: RT = 4.49 min, [MH] - = 603.15.
步骤E：合成(S)-5-(2-(2-((5-氯-2-((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚1,2-二羧酸二叔丁酯Step E: Synthesis of (S)-5-(2-(2-((5-chloro-2-((diallylamino)methyl)phenyl)amino)-2-oxoacetamido)- 3-Phenylpropionamido)-1H-indole 1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000384
Figure PCTCN2020133493-appb-000384
将(S)-5-(2-(2-((5-氯-2-((二烯丙基氨基)甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚1,2-二羧酸二叔丁酯(180毫克,0.27毫摩尔)和1,3-二甲基巴比妥酸(220毫克,1.4毫摩尔)溶于二氯甲烷(20.0毫升)中。氮气保护后,加入四三苯基膦钯(69毫克,0.06毫摩尔),在60摄氏度下搅拌12小时。(S)-5-(2-(2-((5-chloro-2-((diallylamino)methyl)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido)-1H-indole 1,2-dicarboxylic acid di-tert-butyl ester (180 mg, 0.27 mmol) and 1,3-dimethylbarbituric acid (220 mg, 1.4 mmol) In dichloromethane (20.0 mL). After nitrogen protection, palladium tetrakistriphenylphosphine (69 mg, 0.06 mmol) was added and stirred at 60 degrees Celsius for 12 hours.
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得到86毫克黄色固体(S)-5-(2-(2-(((2-(氨基甲基)-5-氯苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯盐(收率：53％)和55毫克黄色固体(S)-5-(2-(2-(((2-(氨基甲基)-5-羟基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)二叔丁基)-1H-吲哚-1,2-二羧酸盐(收率：34％)。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 to obtain 86 mg of yellow solid (S)-5-(2-(2-(((2-(aminomethyl)-5-chlorophenyl)amino)-2-oxo Acetamido)-3-phenylpropionamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester salt (yield: 53%) and 55 mg of yellow solid (S)-5-( 2-(2-(((2-(Aminomethyl)-5-hydroxyphenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)di-tert-butyl)-1H- Indole-1,2-dicarboxylate (yield: 34%).
步骤F：合成(S)-5-(2-(2-(((2-(氨基甲基)-5-氯苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Step F: Synthesis of (S)-5-(2-(2-(((2-(aminomethyl)-5-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamide Base)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000385
Figure PCTCN2020133493-appb-000385
将(S)-5-(2-(2-(((2-(氨基甲基)-5-氯苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)二叔丁基)-1H-吲哚-1,2-二羧酸盐(86毫克,0.09毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((2-(aminomethyl)-5-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) Di-tert-butyl)-1H-indole-1,2-dicarboxylate (86 mg, 0.09 mmol) was dissolved in dichloromethane (2.0 mL). Subsequently, trifluoroacetic acid (0.5 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38毫克白色固体(S)-5-(2-(2-(((2-(氨基甲基)-5-氯苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：57％)。LCMS：RT＝2.86min,[M-H] -＝533.15。 1H NMR(400MHz,DMSO)δ12.95(s,1H),11.75(s,1H),10.59(s,1H),10.12(s,1H),8.89(d,J＝8.6Hz,1H),8.08(s,2H),7.98(d,J＝1.9Hz,1H),7.54(dd,J＝5.3,3.1Hz,1H),7.48(dd,J＝8.4,2.2Hz,1H),7.40(d,J＝8.8Hz,1H),7.33(ddd,J＝20.1,9.7,4.7Hz,3H),7.23(dt,J＝9.4,4.2Hz,1H),7.07(dd,J＝2.1,0.8Hz,1H),4.86–4.73(m,1H),3.96(d,J＝5.0Hz,1H),3.22–3.17(m,1H),2.55(s,1H),2.53–2.52(m,1H)。 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 38 mg of white solid (S)-5-(2-(2-(((2-(aminomethyl)-5-chlorophenyl)amino)- 2-oxoacetamido)-3-phenylpropionamido)-1H-indole-2-carboxylic acid (yield: 57%). LCMS: RT = 2.86 min, [MH] - =533.15. 1 H NMR (400MHz, DMSO) δ 12.95 (s, 1H), 11.75 (s, 1H), 10.59 (s, 1H), 10.12 (s, 1H), 8.89 (d, J = 8.6 Hz, 1H), 8.08 ( s, 2H), 7.98 (d, J = 1.9 Hz, 1H), 7.54 (dd, J = 5.3, 3.1 Hz, 1H), 7.48 (dd, J = 8.4, 2.2 Hz, 1H), 7.40 (d, J =8.8Hz, 1H), 7.33 (ddd, J = 20.1, 9.7, 4.7 Hz, 3H), 7.23 (dt, J = 9.4, 4.2 Hz, 1H), 7.07 (dd, J = 2.1, 0.8 Hz, 1H) , 4.86–4.73(m,1H), 3.96(d,J=5.0Hz,1H), 3.22–3.17(m,1H), 2.55(s,1H), 2.53–2.52(m,1H).
实施例66Example 66
合成(S)-5-(2-(2-(((2-(氨基甲基)-5-羟基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-(((2-(aminomethyl)-5-hydroxyphenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)- 1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000386
Figure PCTCN2020133493-appb-000386
具体合成路线如下：The specific synthesis route is as follows:
步骤F：合成(S)-5-(2-(2-(((2-(氨基甲基)-5-羟基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Step F: Synthesis of (S)-5-(2-(2-(((2-(aminomethyl)-5-hydroxyphenyl)amino)-2-oxoacetamido)-3-phenylpropionamide Base)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000387
Figure PCTCN2020133493-appb-000387
将(S)-5-(2-(2-(((2-(氨基甲基)-5-羟基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(55毫克,0.08毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-5-(2-(2-(((2-(aminomethyl)-5-hydroxyphenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) -1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (55 mg, 0.08 mmol) was dissolved in dichloromethane (2.0 mL). Then, trifluoroacetic acid (0.5 mL) was added to the above solution , Stir 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21毫克白色固体(S)-5-(2-(2-(((2-(氨基甲基)-5-羟基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：50％)。LCMS：RT＝3.75min,[M-H] -＝514.09。 1H NMR(400MHz,DMSO)δ13.08–12.75(m,1H),11.74(s,1H),10.16(s,1H),9.03(s,1H),7.94(d,J＝1.8Hz,1H),7.38(d,J＝8.9Hz,1H),7.29(dd,J＝8.2,3.1Hz,5H),7.25–7.10(m,4H),7.06(d,J＝1.4Hz,1H),5.76(s,1H),4.86(dd,J＝14.1,8.0Hz,1H),4.72(s,2H),3.14(ddd,J＝21.7,13.7,7.1Hz,3H),2.54–2.52(m,1H)。 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 21 mg of white solid (S)-5-(2-(2-(((2-(aminomethyl)-5-hydroxyphenyl)amino)- 2-oxoacetamido)-3-phenylpropionamido)-1H-indole-2-carboxylic acid (yield: 50%). LCMS: RT = 3.75 min, [MH] - =514.09. 1 H NMR (400MHz, DMSO) δ 13.08-12.75 (m, 1H), 11.74 (s, 1H), 10.16 (s, 1H), 9.03 (s, 1H), 7.94 (d, J = 1.8 Hz, 1H), 7.38 (d, J = 8.9 Hz, 1H), 7.29 (dd, J = 8.2, 3.1 Hz, 5H), 7.25-7.10 (m, 4H), 7.06 (d, J = 1.4 Hz, 1H), 5.76 (s , 1H), 4.86 (dd, J = 14.1, 8.0 Hz, 1H), 4.72 (s, 2H), 3.14 (ddd, J = 21.7, 13.7, 7.1 Hz, 3H), 2.54-2.52 (m, 1H).
实施例67Example 67
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((N-ethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000388
Figure PCTCN2020133493-appb-000388
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((N-乙基氨磺酰基)氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((N-ethylsulfamoyl)amino)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000389
Figure PCTCN2020133493-appb-000389
0摄氏度下,将(S)-5-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(100毫克,0.16毫摩尔)和N,N-二异丙基乙胺(0.3毫升,2.4毫摩尔)溶于四氢呋喃(6.0毫升)中。随后,向上述溶液中加乙基氨磺酰氯(30毫克,0.2毫摩尔)。在0摄氏度反应1小时。At 0 degrees Celsius, (S)-5-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino) -2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (100mg, 0.16mmol) and N,N-diisopropylethylamine (0.3ml, 2.4 (Mmol) was dissolved in tetrahydrofuran (6.0 mL). Subsequently, ethyl sulfamoyl chloride (30 mg, 0.2 mmol) was added to the above solution. React at 0 degrees Celsius for 1 hour.
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,所得残余物用硅胶柱层析纯化得到80毫克黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((N-乙基氨磺酰基)氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：69％)。LCMS：RT＝3.96min,[M-H] -＝749.13。 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 The resulting residue was purified by silica gel column chromatography to obtain 80 mg of yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino) -2-oxoacetamido)-3-(4-((N-ethylsulfamoyl)amino)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (Yield: 69%). LCMS: RT = 3.96 min, [MH] - =749.13.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((N-ethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000390
Figure PCTCN2020133493-appb-000390
将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((N-乙基氨磺酰基)氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(80毫克,0.1毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。Add (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((N-ethylsulfamoyl)amino)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (80mg, 0.1mmol) dissolved in dichloromethane (2.0ml) Afterwards, trifluoroacetic acid (0.5 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51毫克黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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：68％)。LCMS：RT＝3.53min,[M+H] +＝695.11。 1H NMR(400MHz,DMSO)δ13.15–12.80(m,1H),11.73(s,1H),10.73(s,1H),10.03(s,1H),9.85(s,1H),9.56(s,1H),8.82(d,J＝8.3Hz,1H),8.06–7.89(m,2H),7.76(t,J＝11.1Hz,1H),7.62(dd,J＝8.7,2.3Hz,1H),7.33(ddd,J＝13.2,10.8,5.4Hz,3H),7.17(d,J＝8.5Hz,2H),7.06(dd,J＝5.4,3.2Hz,3H),5.76(s,1H),4.67(dd,J＝14.6,7.6Hz,1H),3.09(d,J＝6.3Hz,2H),2.88–2.74(m,2H),2.53–2.52(m,1H),1.30–1.22(m,1H),0.92(t,J＝7.2Hz,3H)。 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 51 mg of yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-((N-ethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield : 68%). LCMS: RT = 3.53 min, [M+H] + =695.11. 1 H NMR (400MHz, DMSO) δ 13.15-12.80 (m, 1H), 11.73 (s, 1H), 10.73 (s, 1H), 10.03 (s, 1H), 9.85 (s, 1H), 9.56 (s ,1H),8.82(d,J=8.3Hz,1H),8.06-7.89(m,2H),7.76(t,J=11.1Hz,1H),7.62(dd,J=8.7,2.3Hz,1H) ,7.33(ddd,J=13.2,10.8,5.4Hz,3H), 7.17(d,J=8.5Hz,2H), 7.06(dd,J=5.4,3.2Hz,3H), 5.76(s,1H), 4.67(dd,J=14.6,7.6Hz,1H), 3.09(d,J=6.3Hz,2H), 2.88–2.74(m,2H), 2.53–2.52(m,1H), 1.30–1.22(m, 1H), 0.92 (t, J=7.2 Hz, 3H).
实施例68Example 68
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((N,N-Dimethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000391
Figure PCTCN2020133493-appb-000391
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((N，N-二甲基氨磺酰基)氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((N,N-Dimethylsulfamoyl)amino)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000392
Figure PCTCN2020133493-appb-000392
0摄氏度下,将(S)-5-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(100毫克,0.16毫摩尔)和N,N-二异丙基乙胺(0.3毫升,2.4毫摩尔)溶于四氢呋喃(6.0毫升)中。随后,向上述溶液中加N，N-二甲基氨磺酰氯(30毫克,0.2毫摩尔)。在0℃反应1小时。At 0 degrees Celsius, (S)-5-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino) -2-oxoacetamido) propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (100 mg, 0.16 mmol) and N,N-diisopropylethylamine (0.3 ml, 2.4 (Mmol) was dissolved in tetrahydrofuran (6.0 mL). Subsequently, N,N-dimethylsulfamoyl chloride (30 mg, 0.2 mmol) was added to the above solution. React at 0°C for 1 hour.
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,所得残余物用硅胶柱层析纯化得到55毫克黄色固体叔丁基(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((N，N-二甲基氨磺酰基)氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸酯(收率：69％)。RT＝4.02min,[M-H] -＝749.13。 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 The resulting residue was purified by silica gel column chromatography to obtain 55 mg of yellow solid tert-butyl (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-((N,N-dimethylsulfamoyl)amino)phenylpropionamido)-1H-indole-2-carboxylate (Yield: 69%) RT = 4.02 min, [MH] - =749.13.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((N,N-Dimethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000393
Figure PCTCN2020133493-appb-000393
将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((N，N-二甲基氨磺酰基)氨基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(55毫克,0.07毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。Add (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((N,N-Dimethylsulfamoyl)amino)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (55 mg, 0.07 mmol) dissolved in dichloromethane (2.0 ML). Then, trifluoroacetic acid (0.5 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11毫克黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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：22％)。LCMS：RT＝3.57min,[M-H] -＝693.04。 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 11 mg of yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-((N,N-dimethylsulfamoyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid (Yield: 22%). LCMS: RT = 3.57 min, [MH] - =693.04.
实施例69Example 69
合成4-((S)-2-(2-((5-氯-2-((R)-3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of 4-((S)-2-(2-((5-chloro-2-((R)-3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido) -3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000394
Figure PCTCN2020133493-appb-000394
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(R)-1-(4-氯-2-硝基苯基)吡咯烷-3-醇Step A: Synthesis of (R)-1-(4-chloro-2-nitrophenyl)pyrrolidin-3-ol
Figure PCTCN2020133493-appb-000395
Figure PCTCN2020133493-appb-000395
将4-氯-1-氟-2-硝基苯(1.75克,10.0毫摩尔),(R)-吡咯烷丁-3-醇(1.0毫升,36.2毫摩尔)和碳酸铯(6.5克,20.0毫摩尔)溶于N,N-二甲基甲酰胺(3.0毫升)中。加热升温到50℃并恒温搅拌5小时后,LCMS监测至反应完全,将反应液冷却至室温。Combine 4-chloro-1-fluoro-2-nitrobenzene (1.75 g, 10.0 mmol), (R)-pyrrolidin-3-ol (1.0 mL, 36.2 mmol) and cesium carbonate (6.5 g, 20.0 (Mmol) was dissolved in N,N-dimethylformamide (3.0 mL). After heating to 50°C and stirring at constant temperature for 5 hours, LCMS monitored until the reaction was complete, and the reaction solution was cooled to room temperature.
将反应液缓慢滴加到水(15.0毫升)中,混合液用乙酸乙酯(10毫升×3次)萃取,合并有机相,有机相用饱和食盐水(10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得到2.2克黄色固体(R)-1-(4-氯-2-硝基苯基)吡咯烷-3-醇(收率：92.0％)。The reaction solution was slowly added dropwise to water (15.0 ml), the mixed solution was extracted with ethyl acetate (10 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (10 ml × 3 times), anhydrous sulfuric acid Dry with sodium and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 2.2 g of yellow solid (R)-1-(4-chloro-2-nitrophenyl)pyrrolidin-3-ol (yield: 92.0%).
步骤B：合成(R)-1-(2-氨基-4-氯苯基)吡咯烷-3-醇Step B: Synthesis of (R)-1-(2-amino-4-chlorophenyl)pyrrolidin-3-ol
Figure PCTCN2020133493-appb-000396
Figure PCTCN2020133493-appb-000396
将(R)-1-(4-氯-2-硝基苯基)吡咯烷-3-醇(2.2克,9.1毫摩尔)和醋酸(1.0毫升)溶于甲醇(15.0毫升)中，然后将铁粉(5.1克,91.0毫摩尔)加入反应液中,在50℃下搅拌2小时,LCMS监测至反应完全。(R)-1-(4-chloro-2-nitrophenyl)pyrrolidin-3-ol (2.2 g, 9.1 mmol) and acetic acid (1.0 mL) were dissolved in methanol (15.0 mL), and then Iron powder (5.1 g, 91.0 mmol) was added to the reaction solution and stirred at 50°C for 2 hours. LCMS monitored until the reaction was complete.
反应液冷却至室温,垫硅藻土过滤,滤饼用乙酸乙酯(10毫升×3次)洗涤,合并滤液及洗涤液,有机相经水和饱和食盐水各洗一次,减压浓缩，残渣经硅胶柱层析纯化得到1.9克黄色固体(R)-1-(2-氨基-4-氯苯基)吡咯烷-3-醇(收率：32.0％)。 LCMS：RT＝2.84min,[M+H] +＝213.07。 The reaction solution was cooled to room temperature, filtered through a pad of celite, the filter cake was washed with ethyl acetate (10 ml×3 times), the filtrate and washing liquid were combined, the organic phase was washed once with water and saturated brine, and concentrated under reduced pressure. The residue Purified by silica gel column chromatography to obtain 1.9 g of yellow solid (R)-1-(2-amino-4-chlorophenyl)pyrrolidin-3-ol (yield: 32.0%). LCMS: RT = 2.84 min, [M+H] + =213.07.
步骤C：合成(R)-2-(((5-氯-2-(3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酸甲酯Step C: Synthesis of (R)-2-(((5-chloro-2-(3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoacetate methyl ester
Figure PCTCN2020133493-appb-000397
Figure PCTCN2020133493-appb-000397
氮气保护下,将(R)-1-(2-氨基-4-氯苯基)吡咯烷-3-醇(1.9克,9.0毫摩尔)和吡啶(1.8毫升,18.0毫摩尔)溶于二氯甲烷(20.0毫升)中,冰水浴中搅拌5分钟,向反应液中缓慢滴加草酰氯单甲酯(1.7毫升,10.7毫摩尔)的二氯甲烷溶液(3.0毫升),冰水浴中搅拌1小时。Under nitrogen protection, dissolve (R)-1-(2-amino-4-chlorophenyl)pyrrolidin-3-ol (1.9 g, 9.0 mmol) and pyridine (1.8 mL, 18.0 mmol) in dichloromethane In methane (20.0 ml), stir in an ice-water bath for 5 minutes, slowly add dropwise a dichloromethane solution (3.0 ml) of monomethyl oxalyl chloride (1.7 ml, 10.7 mmol) to the reaction solution, and stir in an ice-water bath for 1 hour .
向反应液中加水(10.0毫升)淬灭反应,用乙酸乙酯(10毫升×3次)萃取。合并有机相,减压浓缩,所得残余物用硅胶柱层析纯化得到2.4克黄色固体(R)-2-(((5-氯-2-(3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酸甲酯(收率：90.0％)。LCMS：RT＝3.46min,[M+H] +＝299.07。 Water (10.0 mL) was added to the reaction solution to quench the reaction, and it was extracted with ethyl acetate (10 mL×3 times). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain 2.4 g of yellow solid (R)-2-(((5-chloro-2-(3-hydroxypyrrolidin-1-yl)phenyl) ) Methyl amino)-2-oxoacetate (yield: 90.0%). LCMS: RT = 3.46 min, [M+H] + =299.07.
步骤D：合成(R)-2-(((5-氯-2-(3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酸Step D: Synthesis of (R)-2-(((5-chloro-2-(3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000398
Figure PCTCN2020133493-appb-000398
将(R)-2-(((5-氯-2-(3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酸甲酯(2.4克,8.0毫摩尔)溶于四氢呋喃(18.0毫升)和水(6.0毫升)中。随后,向上述溶液中加入氢氧化锂一水合物(320毫克,8.0毫摩尔)。在室温下搅拌1小时。(R)-2-(((5-Chloro-2-(3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoacetate methyl ester (2.4 g, 8.0 mmol) was dissolved in Tetrahydrofuran (18.0 mL) and water (6.0 mL). Then, lithium hydroxide monohydrate (320 mg, 8.0 mmol) was added to the above solution. Stir at room temperature for 1 hour.
加稀盐酸调节pH至弱酸性,减压浓缩除去大部分溶剂,加稀盐酸调节pH至3,析出大量白色固体,过滤,收集滤饼得到2.0克黄色固体(R)-2-(((5-氯-2-(3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酸(收率：63％)。LCMS：RT＝4.04min,[M+H] +＝285.07。 Add dilute hydrochloric acid to adjust the pH to weak acidity, concentrate under reduced pressure to remove most of the solvent, add dilute hydrochloric acid to adjust the pH to 3, precipitate a large amount of white solid, filter, and collect the filter cake to obtain 2.0 g of yellow solid (R)-2-(((5 -Chloro-2-(3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoacetic acid (Yield: 63%). LCMS: RT = 4.04 min, [M+H] + =285.07 .
步骤E：合成4-((S)-2-(2-((5-氯-2-((R)-3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of 4-((S)-2-(2-((5-chloro-2-((R)-3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000399
Figure PCTCN2020133493-appb-000399
将(R)-2-(((5-氯-2-(3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酸(680毫克,2.4毫摩尔),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(1.4克,3.6毫摩尔)和N,N-二异丙基乙胺(1.2毫升,7.2毫摩尔)溶于N,N-二甲基甲酰胺(20.0毫升)中。随后,向上述溶液中加入(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(1.0克,3.0毫摩尔),在室温下搅拌1小时。(R)-2-(((5-chloro-2-(3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoacetic acid (680 mg, 2.4 mmol), 2-( 7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.4g, 3.6mmol) and N,N-diisopropylethylamine (1.2 Ml, 7.2 mmol) was dissolved in N,N-dimethylformamide (20.0 ml). Then, (S)-4-(2-amino-3-phenylpropionamido)benzene was added to the above solution Tert-Butyl formate (1.0 g, 3.0 mmol), stirred at room temperature for 1 hour.
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得到960毫克黄色固体叔丁基4-((S)-2-(2-((5-氯-2-((R)-3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸酯(收率：66.0％)。LCMS：RT＝4.52min,[M+H] +＝607.08。 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 to obtain 960 mg of yellow solid tert-butyl 4-((S)-2-(2-((5-chloro-2-((R)-3-hydroxypyrrolidine-1- (Phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoate (yield: 66.0%). LCMS: RT=4.52 min, [M+H] + =607.08.
步骤F：合成4-((S)-2-(2-((5-氯-2-((R)-3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of 4-((S)-2-(2-((5-chloro-2-((R)-3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoethyl Amido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000400
Figure PCTCN2020133493-appb-000400
将4-((S)-2-(2-((5-氯-2-((R)-3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(80毫克,0.13毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。Add 4-((S)-2-(2-((5-chloro-2-((R)-3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido) Tert-butyl-3-phenylpropionamido)benzoate (80 mg, 0.13 mmol) was dissolved in dichloromethane (2.0 mL). Subsequently, trifluoroacetic acid (0.5 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13毫克白色固体4-((S)-2-(2-((5-氯-2-((R)-3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：18.0％)。LCMS：RT＝3.92min,[M-H] -＝549.09。 1H NMR(400MHz,DMSO)δ13.05–12.62(m,1H),10.49(s,1H),10.01(s,1H),9.02(d,J＝8.4Hz,1H),8.01–7.88(m,2H),7.81–7.66(m,2H),7.55(d,J＝2.6Hz,1H),7.37–7.25(m,3H),7.21(dt,J＝9.3,4.2Hz,1H),7.15(dd,J＝8.8,2.6Hz,1H),6.95(d,J＝8.9Hz,1H),4.83–4.73(m,1H),4.36–4.25(m,1H),3.38–3.12(m,3H),3.05(ddd,J＝12.9,8.5,4.2Hz,1H),2.87(dd,J＝9.9,2.1Hz,1H),1.95(dt,J＝13.4,7.7Hz,1H),1.83–1.71(m,1H)。 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 13 mg of white solid 4-((S)-2-(2-((5-chloro-2-((R)-3-hydroxypyrrolidine-1) -Yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid (yield: 18.0%). LCMS: RT = 3.92 min, [MH] - =549.09. 1 H NMR (400MHz, DMSO) δ 13.05-12.62 (m, 1H), 10.49 (s, 1H), 10.01 (s, 1H), 9.02 (d, J = 8.4 Hz, 1H), 8.01-7.88 (m ,2H),7.81-7.66(m,2H),7.55(d,J=2.6Hz,1H),7.37-7.25(m,3H),7.21(dt,J=9.3,4.2Hz,1H),7.15( dd,J=8.8,2.6Hz,1H), 6.95(d,J=8.9Hz,1H), 4.83–4.73(m,1H), 4.36–4.25(m,1H), 3.38–3.12(m,3H) ,3.05(ddd,J＝12.9,8.5,4.2Hz,1H), 2.87(dd,J＝9.9,2.1Hz,1H),1.95(dt,J＝13.4,7.7Hz,1H),1.83–1.71(m ,1H).
实施例70Example 70
合成(S)-4-(2-(2-((5-氯-2-(3-氧吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(3-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) benzoic acid
Figure PCTCN2020133493-appb-000401
Figure PCTCN2020133493-appb-000401
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-((S)-2-(2-((5-氯-2-(3-氧吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of 4-((S)-2-(2-((5-chloro-2-(3-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido)- 3-phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000402
Figure PCTCN2020133493-appb-000402
将4-((S)-2-(2-((5-氯-2-((R)-3-羟基吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(350毫克,0.6毫摩尔),碳酸氢钠(100毫克,1.2毫摩尔)溶于二氯甲烷(13.0毫升)中,冰水浴中搅拌10分钟后,加戴斯-马丁试剂(1.0克,2.4毫摩尔),室温反应1小时。Add 4-((S)-2-(2-((5-chloro-2-((R)-3-hydroxypyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido) -3-Phenylpropionamido) tert-butyl benzoate (350 mg, 0.6 mmol), sodium bicarbonate (100 mg, 1.2 mmol) dissolved in dichloromethane (13.0 ml), stirred in an ice-water bath for 10 Minutes later, add Dess-Martin reagent (1.0 g, 2.4 mmol) and react at room temperature for 1 hour.
将反应液缓慢滴加到水(15.0毫升)中,混合液用乙酸乙酯(10毫升×3次)萃取,合并有机相,有机相用饱和食盐水(10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得到280毫克黄色固体4-((S)-2-(2-((5-氯-2-(3-氧吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：80.0％)。LCMS：RT＝4.52min,[M+H] +＝605.13。 The reaction solution was slowly added dropwise to water (15.0 ml), the mixed solution was extracted with ethyl acetate (10 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (10 ml × 3 times), anhydrous sulfuric acid Dry with sodium and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 280 mg of yellow solid 4-((S)-2-(2-((5-chloro-2-(3-oxopyrrolidin-1-yl)phenyl)amino) -2-oxoacetamido)-3-phenylpropionamido)tert-butyl benzoate (yield: 80.0%). LCMS: RT = 4.52 min, [M+H] + =605.13.
步骤B：合成4-((S)-2-(2-((5-氯-2-(3-氧吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step B: Synthesis of 4-((S)-2-(2-((5-chloro-2-(3-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido)- 3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000403
Figure PCTCN2020133493-appb-000403
将4-((S)-2-(2-((5-氯-2-(3-氧吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(80毫克,0.13毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。Add 4-((S)-2-(2-((5-chloro-2-(3-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene Tert-butyl propionamido)benzoate (80 mg, 0.13 mmol) was dissolved in dichloromethane (2.0 mL). Subsequently, trifluoroacetic acid (0.5 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13毫克白色固体4-((S)-2-(2-((5-氯-2-(3-氧吡咯烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：18.0％)。LCMS：RT＝4.07min,[M-H] - ＝548.15。 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 13 mg of white solid 4-((S)-2-(2-((5-chloro-2-(3-oxopyrrolidin-1-yl)benzene) (Yl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid (yield: 18.0%). LCMS: RT = 4.07 min, [MH] - =548.15.
实施例71Example 71
合成(S)-4-(2-(2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetamido)-3- Phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000404
Figure PCTCN2020133493-appb-000404
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成3-(4-氯-2-硝基苯基)恶唑烷-2-酮Step A: Synthesis of 3-(4-chloro-2-nitrophenyl)oxazolidin-2-one
Figure PCTCN2020133493-appb-000405
Figure PCTCN2020133493-appb-000405
将4-氯-1-氟-2-硝基苯(0.5克,2.9毫摩尔),恶唑烷丁-2-酮(500毫克,5.7毫摩尔)和碳酸钾(790毫克,5.7毫摩尔)溶于N,N-二甲基甲酰胺(3.0毫升)中。加热升温到50℃并恒温搅拌15小时后,LCMS监测至反应完全,将反应液冷却至室温。Combine 4-chloro-1-fluoro-2-nitrobenzene (0.5 g, 2.9 mmol), oxazolidin-2-one (500 mg, 5.7 mmol) and potassium carbonate (790 mg, 5.7 mmol) Dissolve in N,N-dimethylformamide (3.0 mL). After heating to 50°C and stirring at constant temperature for 15 hours, LCMS monitored until the reaction was complete, and the reaction solution was cooled to room temperature.
将反应液缓慢滴加到水(15.0毫升)中,混合液用乙酸乙酯(10毫升×3次)萃取,合并有机相,有机相用饱和食盐水(10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得到0.6克黄色固体3-(4-氯-2-硝基苯基)恶唑烷-2-酮(收率：92.0％)。The reaction solution was slowly added dropwise to water (15.0 ml), the mixed solution was extracted with ethyl acetate (10 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (10 ml × 3 times), anhydrous sulfuric acid Dry with sodium and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 0.6 g of yellow solid 3-(4-chloro-2-nitrophenyl)oxazolidin-2-one (yield: 92.0%).
步骤B：合成3-(4-氯-2-氨基苯基)恶唑烷-2-酮Step B: Synthesis of 3-(4-chloro-2-aminophenyl)oxazolidin-2-one
Figure PCTCN2020133493-appb-000406
Figure PCTCN2020133493-appb-000406
将3-(4-氯-2-硝基苯基)恶唑烷-2-酮(0.5克,2.0毫摩尔)和醋酸(1.0毫升)溶于甲醇(15.0毫升)中，然后将铁粉(560毫克,10.0毫摩尔)加入反应液中,在50℃下搅拌2小时,LCMS监测至反应完全。Dissolve 3-(4-chloro-2-nitrophenyl)oxazolidin-2-one (0.5 g, 2.0 mmol) and acetic acid (1.0 mL) in methanol (15.0 mL), and then mix the iron powder ( 560 mg, 10.0 mmol) was added to the reaction solution, stirred at 50°C for 2 hours, LCMS monitored until the reaction was complete.
反应液冷却至室温,垫硅藻土过滤,滤饼用乙酸乙酯(10毫升×3次)洗涤,合并滤液及洗涤液,有机相经水和饱和食盐水各洗一次,减压浓缩，残渣经硅胶柱层析纯化得到0.4克黄色液体3-(4-氯-2-氨基苯基)恶唑烷-2-酮(收率：92.0％)。The reaction solution was cooled to room temperature, filtered through a pad of celite, the filter cake was washed with ethyl acetate (10 ml×3 times), the filtrate and washing liquid were combined, the organic phase was washed once with water and saturated brine, and concentrated under reduced pressure. The residue Purified by silica gel column chromatography to obtain 0.4 g of yellow liquid 3-(4-chloro-2-aminophenyl)oxazolidin-2-one (yield: 92.0%).
步骤C：合成2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000407
Figure PCTCN2020133493-appb-000407
氮气保护下,将(R)-1-(2-氨基-4-氯苯基)吡咯烷-3-醇(0.4克,1.9毫摩尔)和吡啶(0.3毫升,2.8毫摩尔)溶于二氯甲烷(5.0毫升)中,冰水浴中搅拌5分钟,向反应液中缓慢滴加草酰氯单甲酯(280毫克,2.3毫摩尔)的二氯甲烷溶液(3.0毫升),冰水浴中搅拌1小时。Under nitrogen protection, (R)-1-(2-amino-4-chlorophenyl)pyrrolidin-3-ol (0.4 g, 1.9 mmol) and pyridine (0.3 mL, 2.8 mmol) were dissolved in dichloromethane In methane (5.0 ml), stir in an ice-water bath for 5 minutes, slowly add dropwise a dichloromethane solution (3.0 ml) of monomethyl oxalyl chloride (280 mg, 2.3 mmol) to the reaction solution, and stir in an ice-water bath for 1 hour .
向反应液中加水(10.0毫升)淬灭反应,用乙酸乙酯(10毫升×3次)萃取。合并有机相,减压浓缩,所得残余物用硅胶柱层析纯化得到560毫克黄色固体2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酸甲酯(收率：100.0％)。LCMS：RT＝3.30min,[M-H] -＝298.99。 Water (10.0 mL) was added to the reaction solution to quench the reaction, and it was extracted with ethyl acetate (10 mL×3 times). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain 560 mg of yellow solid 2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino) Methyl-2-oxoacetate (yield: 100.0%). LCMS: RT = 3.30 min, [MH] - =298.99.
步骤D：合成2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酸Step D: Synthesis of 2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000408
Figure PCTCN2020133493-appb-000408
将2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酸甲酯(560毫克,1.9毫摩尔)溶于四氢呋喃(12.0毫升)和水(4.0毫升)中。随后,向上述溶液中加入氢氧化锂一水合物(76毫克,1.9毫摩尔)。在室温下搅拌1小时。Dissolve 2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetate methyl ester (560 mg, 1.9 mmol) in tetrahydrofuran (12.0 ml ) And water (4.0 ml). Subsequently, lithium hydroxide monohydrate (76 mg, 1.9 mmol) was added to the above solution. Stir at room temperature for 1 hour.
加稀盐酸调节pH至弱酸性,减压浓缩除去大部分溶剂,加稀盐酸调节pH至3,析出大量白色固体,过滤,收集滤饼得到0.5克黄色固体2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酸(收率：93％)。LCMS：RT＝3.69min,[M-H] -＝282.92。 Add dilute hydrochloric acid to adjust the pH to weak acidity, concentrate under reduced pressure to remove most of the solvent, add dilute hydrochloric acid to adjust the pH to 3, precipitate a large amount of white solid, filter, collect the filter cake to obtain 0.5 g of yellow solid 2-((5-chloro-2- (2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetic acid (yield: 93%). LCMS: RT = 3.69 min, [MH] - = 282.92.
步骤E：合成(S)-4-(2-(2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetamido )-3-Phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000409
Figure PCTCN2020133493-appb-000409
将2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酸(100毫克,0.35毫摩尔),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(0.2克,0.5毫摩尔)和N,N-二异丙基乙胺(0.2毫升,1.2毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)中。随后,向上述溶液中加入(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(144毫克,0.42毫摩尔),在室温下搅拌1小时。Add 2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetic acid (100 mg, 0.35 mmol), 2-(7-azo Benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.2g, 0.5mmol) and N,N-diisopropylethylamine (0.2ml, 1.2ml) Mol) was dissolved in N,N-dimethylformamide (5.0 mL). Subsequently, tert-butyl (S)-4-(2-amino-3-phenylpropionamido)benzoate (144 mg, 0.42 mmol) was added to the above solution, and stirred at room temperature for 1 hour.
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得到150毫克黄色固体(S)-4-(2-(2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：66.0％)。LCMS：RT＝4.36min,[M-H] -＝605.14。 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 to obtain 150 mg of yellow solid (S)-4-(2-(2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl) Amino)-2-oxoacetamido)-3-phenylpropionamido)tert-butyl benzoate (yield: 66.0%). LCMS: RT = 4.36 min, [MH] - =605.14.
步骤F：合成(S)-4-(2-(2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetamido) -3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000410
Figure PCTCN2020133493-appb-000410
将(S)-4-(2-(2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(150毫克,0.25毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-4-(2-(2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido) tert-butyl benzoate (150 mg, 0.25 mmol) was dissolved in dichloromethane (2.0 mL). Subsequently, trifluoroacetic acid (0.5 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69毫克白色固体(S)-4-(2-(2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：51.0％)。LCMS：RT＝3.80min,[M-H] -＝549.01。 1H NMR(400MHz,DMSO)δ12.99–12.60(m,1H),10.51(s,1H),10.12(s,1H),9.13(d,J＝8.3Hz,1H),8.00–7.89(m,2H),7.86(d,J＝2.4Hz,1H),7.77–7.67(m,2H),7.53(d,J＝8.7Hz,1H),7.39(dd,J＝8.7,2.5Hz,1H),7.31(dt,J＝14.9,4.6Hz,3H),7.21(t,J＝7.1Hz,1H),4.76(dd,J＝15.2,7.3Hz,1H),4.45(dd,J＝9.6,7.5Hz,2H),3.99(dd,J＝9.4,7.4Hz,2H),3.19(d,J＝7.4Hz,2H),2.53–2.52(m,1H)。 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 69 mg of white solid (S)-4-(2-(2-((5-chloro-2-(2-oxazolidin-3-yl) )Phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid (yield: 51.0%). LCMS: RT = 3.80 min, [MH] - =549.01. 1 H NMR (400MHz, DMSO) δ 12.99-12.60 (m, 1H), 10.51 (s, 1H), 10.12 (s, 1H), 9.13 (d, J = 8.3 Hz, 1H), 8.00-7.89 (m ,2H),7.86(d,J=2.4Hz,1H),7.77–7.67(m,2H),7.53(d,J=8.7Hz,1H),7.39(dd,J=8.7,2.5Hz,1H) ,7.31(dt,J＝14.9,4.6Hz,3H),7.21(t,J＝7.1Hz,1H),4.76(dd,J＝15.2,7.3Hz,1H),4.45(dd,J＝9.6,7.5 Hz, 2H), 3.99 (dd, J = 9.4, 7.4 Hz, 2H), 3.19 (d, J = 7.4 Hz, 2H), 2.53-2.52 (m, 1H).
实施例72Example 72
合成(S)-4-(2-(2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(2-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) benzoic acid
Figure PCTCN2020133493-appb-000411
Figure PCTCN2020133493-appb-000411
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成3-(4-氯-2-硝基苯基)恶唑烷-2-酮Step A: Synthesis of 3-(4-chloro-2-nitrophenyl)oxazolidin-2-one
Figure PCTCN2020133493-appb-000412
Figure PCTCN2020133493-appb-000412
将4-氯-1-氟-2-硝基苯(0.5克,2.9毫摩尔),吡咯烷-2-酮(500毫克,5.7毫摩尔)和碳酸钾(790毫克,5.7毫摩尔)溶于N,N-二甲基甲酰胺(3.0毫升)中。加热升温到50℃并恒温搅拌15小时后,LCMS监测至反应完全,将反应液冷却至室温。Dissolve 4-chloro-1-fluoro-2-nitrobenzene (0.5 g, 2.9 mmol), pyrrolidin-2-one (500 mg, 5.7 mmol) and potassium carbonate (790 mg, 5.7 mmol) N,N-dimethylformamide (3.0 mL). After heating to 50°C and stirring at constant temperature for 15 hours, LCMS monitored until the reaction was complete, and the reaction solution was cooled to room temperature.
将反应液缓慢滴加到水(15.0毫升)中,混合液用乙酸乙酯(10毫升×3次)萃取,合并有机相,有机相用饱和食盐水(10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得到0.6克黄色固体1-(2-硝基-4-氯苯基)吡咯烷-2-酮(收率：92.0％)。The reaction solution was slowly added dropwise to water (15.0 ml), the mixed solution was extracted with ethyl acetate (10 ml × 3 times), the organic phases were combined, and the organic phase was saturated brine (10 ml × 3 times), anhydrous sulfuric acid Dry with sodium and concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography to obtain 0.6 g of yellow solid 1-(2-nitro-4-chlorophenyl)pyrrolidin-2-one (yield: 92.0%).
步骤B：合成1-(2-氨基-4-氯苯基)吡咯烷-2-酮Step B: Synthesis of 1-(2-amino-4-chlorophenyl)pyrrolidin-2-one
Figure PCTCN2020133493-appb-000413
Figure PCTCN2020133493-appb-000413
将1-(2-硝基-4-氯苯基)吡咯烷-2-酮(0.5克,2.0毫摩尔)和醋酸(1.0毫升)溶于甲醇(15.0毫升)中，然后将铁粉(560毫克,10.0毫摩尔)加入反应液中,在50℃下搅拌2小时,LCMS监测至反应完全。Dissolve 1-(2-nitro-4-chlorophenyl)pyrrolidin-2-one (0.5 g, 2.0 mmol) and acetic acid (1.0 ml) in methanol (15.0 ml), and then mix the iron powder (560 Mg, 10.0 mmol) was added to the reaction solution and stirred at 50°C for 2 hours. LCMS monitored until the reaction was complete.
反应液冷却至室温,垫硅藻土过滤,滤饼用乙酸乙酯(10毫升×3次)洗涤,合并滤液及洗涤液,有机相经水和饱和食盐水各洗一次,减压浓缩，残渣经硅胶柱层析纯化得到0.4克黄色液体1-(2-氨基-4-氯苯基)吡咯烷-2-酮(收率：92.0％)。The reaction solution was cooled to room temperature, filtered through a pad of celite, the filter cake was washed with ethyl acetate (10 ml×3 times), the filtrate and washing liquid were combined, the organic phase was washed once with water and saturated brine, and concentrated under reduced pressure. The residue Purified by silica gel column chromatography to obtain 0.4 g of yellow liquid 1-(2-amino-4-chlorophenyl)pyrrolidin-2-one (yield: 92.0%).
步骤C：合成2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000414
Figure PCTCN2020133493-appb-000414
氮气保护下,将1-(2-氨基-4-氯苯基)吡咯烷-2-酮(0.4克,1.9毫摩尔)和吡啶(0.3毫升,2.8毫摩尔)溶于二氯甲烷(5.0毫升)中,冰水浴中搅拌5分钟,向反应液中缓慢滴加草酰氯单甲酯(280毫克,2.3毫摩尔)的二氯甲烷溶液(3.0毫升),冰水浴中搅拌1小时。Under nitrogen protection, dissolve 1-(2-amino-4-chlorophenyl)pyrrolidin-2-one (0.4 g, 1.9 mmol) and pyridine (0.3 mL, 2.8 mmol) in dichloromethane (5.0 mL). ), stirring in an ice-water bath for 5 minutes, slowly adding dropwise a dichloromethane solution (3.0 ml) of monomethyl oxalyl chloride (280 mg, 2.3 mmol) to the reaction solution, and stirring in an ice-water bath for 1 hour.
向反应液中加水(10.0毫升)淬灭反应,用乙酸乙酯(10毫升×3次)萃取。合并有机相,减压浓缩,所得残余物用硅胶柱层析纯化得到560毫克黄色固体2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧乙酸甲酯(收率：100.0％)。LCMS：RT＝3.34min,[M-H] -＝295.01。 Water (10.0 mL) was added to the reaction solution to quench the reaction, and it was extracted with ethyl acetate (10 mL×3 times). The organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain 560 mg of yellow solid 2-((5-chloro-2-(2-oxopyrrolidin-1-yl)phenyl)amino)- Methyl 2-oxoacetate (yield: 100.0%). LCMS: RT = 3.34 min, [MH] - = 295.01.
步骤D：合成2-((5-氯-2-(2-氧杂恶唑烷-3-基)苯基)氨基)-2-氧乙酸Step D: Synthesis of 2-((5-chloro-2-(2-oxazolidin-3-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000415
Figure PCTCN2020133493-appb-000415
将2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧乙酸甲酯(560毫克,1.9毫摩尔)溶于四氢呋喃(12.0毫升)和水(4.0毫升)中。随后,向上述溶液中加入氢氧化锂一水合物(76毫克,1.9毫摩尔)。在室温下搅拌1小时。Dissolve methyl 2-((5-chloro-2-(2-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetate (560 mg, 1.9 mmol) in tetrahydrofuran (12.0 ml) And water (4.0 ml). Subsequently, lithium hydroxide monohydrate (76 mg, 1.9 mmol) was added to the above solution. Stir at room temperature for 1 hour.
加稀盐酸调节pH至弱酸性,减压浓缩除去大部分溶剂,加稀盐酸调节pH至3,析出大量白色固体,过滤,收集滤饼得到0.5克黄色固体2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧乙酸(收率：93％)。LCMS：RT＝3.93min,[M-H] -＝280.98。 Add dilute hydrochloric acid to adjust the pH to weak acidity, concentrate under reduced pressure to remove most of the solvent, add dilute hydrochloric acid to adjust the pH to 3, precipitate a large amount of white solid, filter, collect the filter cake to obtain 0.5 g of yellow solid 2-((5-chloro-2- (2-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetic acid (yield: 93%). LCMS: RT = 3.93 min, [MH] - =280.98.
步骤E：合成(S)-4-(2-(2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(2-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido) -3-phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000416
Figure PCTCN2020133493-appb-000416
将2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧乙酸(200毫克,0.7毫摩尔),2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(0.4克,1.0毫摩尔)和N,N-二异丙基乙胺(0.3毫升,2.2毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)中。随后,向上述溶液中加入(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(220毫克,0.64毫摩尔),在室温下搅拌1小时。Add 2-((5-chloro-2-(2-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetic acid (200 mg, 0.7 mmol), 2-(7-azobenzene Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.4g, 1.0mmol) and N,N-diisopropylethylamine (0.3ml, 2.2mmol) ) Was dissolved in N,N-dimethylformamide (5.0 mL). Subsequently, tert-butyl (S)-4-(2-amino-3-phenylpropionamido)benzoate (220 mg, 0.64 mmol) was added to the above solution and stirred at room temperature for 1 hour.
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得到150毫克黄色固体(S)-4-(2-(2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：40.0％)。LCMS：RT＝3.88min,[M-H] -＝603.13。 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 to obtain 150 mg of yellow solid (S)-4-(2-(2-((5-chloro-2-(2-oxopyrrolidin-1-yl)phenyl)amino )-2-oxoacetamido)-3-phenylpropionamido)tert-butyl benzoate (yield: 40.0%). LCMS: RT = 3.88 min, [MH] - = 603.13.
步骤F：合成(S)-4-(2-(2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-((5-chloro-2-(2-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido)- 3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000417
Figure PCTCN2020133493-appb-000417
将(S)-4-(2-(2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(150毫克,0.25毫摩尔)溶于二氯甲烷(2.0毫升)中。随后,向上述溶液中加入三氟乙酸(0.5毫升),在室温下搅拌1小时。(S)-4-(2-(2-((5-chloro-2-(2-oxopyrrolidin-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene Tert-butyl propionamido)benzoate (150 mg, 0.25 mmol) was dissolved in dichloromethane (2.0 mL). Subsequently, trifluoroacetic acid (0.5 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43毫克白色固体(S)-4-(2-(2-((5-氯-2-(2-氧吡咯烷基-1-基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：32.0％)。LCMS：RT＝3.84min,[M-H] -＝547.08。 1H NMR(400MHz,DMSO)δ12.85–12.68(m,1H),10.48(s,1H),9.92(s,1H),9.13(d,J＝8.3Hz,1H),7.97–7.87(m,3H),7.77–7.68(m,2H),7.45(d,J＝8.7Hz,1H),7.41–7.25(m,5H),7.22(dt,J＝9.4,4.2Hz,1H),4.75(dd,J＝14.5,8.1Hz,1H),3.79(t,J＝7.3Hz,2H),3.19(d,J＝7.5Hz,2H),2.53–2.52(m,1H),2.44(t,J＝8.5Hz,2H),2.10(dd,J＝14.5,7.4Hz,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 43 mg of white solid (S)-4-(2-(2-((5-chloro-2-(2-oxopyrrolidin-1-yl)) (Phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid (yield: 32.0%). LCMS: RT = 3.84 min, [MH] - =547.08. 1 H NMR (400MHz, DMSO) δ 12.85-12.68 (m, 1H), 10.48 (s, 1H), 9.92 (s, 1H), 9.13 (d, J = 8.3 Hz, 1H), 7.97-7.87 (m ,3H),7.77–7.68(m,2H),7.45(d,J=8.7Hz,1H),7.41–7.25(m,5H),7.22(dt,J=9.4,4.2Hz,1H),4.75( dd, J = 14.5, 8.1 Hz, 1H), 3.79 (t, J = 7.3 Hz, 2H), 3.19 (d, J = 7.5 Hz, 2H), 2.53-2.52 (m, 1H), 2.44 (t, J = 8.5 Hz, 2H), 2.10 (dd, J = 14.5, 7.4 Hz, 2H).
实施例73Example 73
合成(S)-2-(4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯基)乙酸Synthesis of (S)-2-(4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)phenyl)acetic acid
Figure PCTCN2020133493-appb-000418
Figure PCTCN2020133493-appb-000418
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-2-(4-(2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-苯基丙酰胺基)苯基)乙酸叔丁酯Step A: Synthesis of (S)-2-(4-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-phenylpropionamido)phenyl)acetic acid tert Butyl
Figure PCTCN2020133493-appb-000419
Figure PCTCN2020133493-appb-000419
室温条件下，将2-(4-氨基苯基)乙酸叔丁酯(414毫克，2毫摩尔)，(((9H-芴-9-基)甲氧基)羰基)-L-苯丙氨酸(774毫克，2毫摩尔)，2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(1.14克，3毫摩尔)，依次加入25毫升圆底烧瓶中，加入N,N-二甲基甲酰胺4毫升将底物全部溶解，最后加入N,N-二异丙基乙胺(0.99毫升，6毫摩尔)，室温条件下反应12小时。At room temperature, 2-(4-aminophenyl) tert-butyl acetate (414 mg, 2 mmol), (((9H-fluoren-9-yl)methoxy)carbonyl)-L-phenylalanine Acid (774 mg, 2 mmol), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.14 g, 3 mmol), Add to a 25ml round bottom flask in turn, add 4ml N,N-dimethylformamide to dissolve all the substrates, and finally add N,N-diisopropylethylamine (0.99ml, 6mmol) at room temperature. React for 12 hours.
反应结束后，向反应液中加入乙酸乙酯(50毫升)并转移到分液漏斗中，用饱和食盐水洗4次(50毫升×4)，将乙酸乙酯转移到锥形瓶中并加入无水硫酸钠干燥，过滤浓缩后得到油状液体，柱层析(洗脱剂：正己烷/乙酸乙酯＝1/1)纯化后得到白色固体粗品2.0克(S)-2-(4-(2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-苯基丙酰胺基)苯基)乙酸叔丁酯，该粗品直接用于下一步反应。After the completion of the reaction, add ethyl acetate (50 ml) to the reaction solution and transfer to a separatory funnel, wash 4 times with saturated saline (50 ml × 4), transfer ethyl acetate to an Erlenmeyer flask and add water After drying with water sodium sulfate, filtering and concentrating, an oily liquid was obtained. After purification by column chromatography (eluent: n-hexane/ethyl acetate = 1/1), 2.0 g of crude white solid product (S)-2-(4-(2) was obtained. -((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-phenylpropionamido)phenyl)acetic acid tert-butyl ester, the crude product was directly used in the next reaction.
步骤B：合成(S)-2-(4-(2-氨基-3-苯基丙酰胺基)苯基)乙酸叔丁酯Step B: Synthesis of tert-butyl (S)-2-(4-(2-amino-3-phenylpropionamido)phenyl)acetate
Figure PCTCN2020133493-appb-000420
Figure PCTCN2020133493-appb-000420
室温条件下，将(S)-2-(4-(2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-苯基丙酰胺基)苯基)乙酸叔丁酯粗品(2.0克)溶于二氯甲烷(9.3毫升)中，再加入4-甲基哌啶(2.32毫升，19.6毫摩尔)，室温条件下反应12小时，TLC监测反应结束后，加入二氯甲烷(50毫升)稀释反应液并转移到分液漏斗中，用饱和碳酸氢钠溶液洗三次(50毫升*4)，剩余的二氯甲烷转移到锥形瓶中，加入无水硫酸钠干燥，过滤浓缩后得到油状液体，柱层析(洗脱剂：二氯甲烷/甲醇＝20/1)纯化后得到600毫克产物(S)-2-(4-(2-氨基-3-苯基丙酰胺基)苯基)乙酸叔丁酯(收率：86％)。LCMS:RT＝3.09min,[M+H] +＝355.24。 At room temperature, (S)-2-(4-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-phenylpropionamido)phenyl)acetic acid The crude tert-butyl ester (2.0 g) was dissolved in dichloromethane (9.3 ml), and 4-methylpiperidine (2.32 ml, 19.6 mmol) was added. The reaction was carried out at room temperature for 12 hours. After the reaction was monitored by TLC, it was added Dilute the reaction solution with dichloromethane (50ml) and transfer it to a separatory funnel, wash with saturated sodium bicarbonate solution three times (50ml*4), transfer the remaining dichloromethane to an Erlenmeyer flask, add anhydrous sodium sulfate Dry, filter and concentrate to obtain an oily liquid. After purification by column chromatography (eluent: dichloromethane/methanol=20/1), 600 mg of product (S)-2-(4-(2-amino-3-benzene) is obtained. Propyl propionamido) phenyl) acetate (yield: 86%). LCMS: RT = 3.09 min, [M+H] + =355.24.
步骤C：合成(S)-2-(4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯基醋酸叔丁酯Step C: Synthesis of (S)-2-(4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-phenylpropionamido)tert-butyl phenylacetate
Figure PCTCN2020133493-appb-000421
Figure PCTCN2020133493-appb-000421
将2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(91毫克，0.34毫摩尔)，(S)-2-(4-(2-氨基-3-苯基丙酰胺基)苯基)乙酸叔丁酯(121毫克，0.34毫摩尔)，2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(194毫克，0.51毫摩尔)加入烧瓶中，加入N,N-二甲基甲酰胺3毫升将底物全部溶解，最后加入N，N-二异丙基乙胺(0.17毫升，1.0毫摩尔)，室温条件下反应12小时。The 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (91 mg, 0.34 mmol), (S)-2-(4-( 2-Amino-3-phenylpropionamido)phenyl)acetic acid tert-butyl ester (121 mg, 0.34 mmol), 2-(7-benzotriazole oxide)-N,N,N',N' -Tetramethylurea hexafluorophosphate (194 mg, 0.51 mmol) was added to the flask, 3 ml of N,N-dimethylformamide was added to dissolve all the substrate, and finally N,N-diisopropyl ethyl was added Amine (0.17 mL, 1.0 mmol) was reacted at room temperature for 12 hours.
反应结束后，向反应液中加入乙酸乙酯(50毫升)并转移到分液漏斗中，用饱和食盐水洗4次(50毫升*4)，将乙酸乙酯转移到锥形瓶中并加入无水硫酸钠干燥，过滤浓缩后得到油状液体，柱层析(洗脱剂：二氯甲烷/甲醇＝20/1)纯化后得到121毫克产物(S)-2-(4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯基醋酸叔丁酯(收率：59％)。LCMS:RT＝4.19min,[M-H] -＝602.23。 After the reaction is over, add ethyl acetate (50 ml) to the reaction solution and transfer to a separatory funnel, wash 4 times with saturated saline (50 ml*4), transfer ethyl acetate to an Erlenmeyer flask and add water After drying with water sodium sulfate, filtering and concentrating to obtain an oily liquid, column chromatography (eluent: dichloromethane/methanol=20/1) to obtain 121 mg of product (S)-2-(4-(2-(2) -(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)tert-butyl phenylacetate ( Yield: 59%). LCMS: RT = 4.19 min, [MH] - =602.23.
步骤D：合成(S)-2-(4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯基)乙酸Step D: Synthesis of (S)-2-(4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-Phenylpropionamido)phenyl)acetic acid
Figure PCTCN2020133493-appb-000422
Figure PCTCN2020133493-appb-000422
取(S)-2-(4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯基醋酸叔丁酯(100毫克，0.166毫摩尔)加入二氯甲烷(2.5毫升)中，再逐滴加入三氟乙酸(0.5毫升，6.7毫摩尔)，室温条件下反应四小时，反应结束后直接浓缩蒸干得到白色固体，白色固体用二氯甲烷洗三次，过滤后得到26毫克产物，二氯甲烷母液浓缩后得到40毫克粗品。LCMS:RT＝3.64min,[M-H] -＝546.18。 Take (S)-2-(4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-Phenylpropionamido)-tert-butyl phenylacetate (100 mg, 0.166 mmol) was added to dichloromethane (2.5 mL), and then trifluoroacetic acid (0.5 mL, 6.7 mmol) was added dropwise at room temperature. After reacting for four hours, after the reaction, it was directly concentrated and evaporated to dryness to obtain a white solid. The white solid was washed three times with dichloromethane, and 26 mg of product was obtained after filtration. The dichloromethane mother liquor was concentrated to obtain 40 mg of crude product. LCMS:RT=3.64min, [MH] - = 546.18.
实施例74Example 74
合成(S)-4-(2-(2-((5-氯-2-(甲基氨基甲酰基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(methylcarbamoyl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido )benzoic acid
Figure PCTCN2020133493-appb-000423
Figure PCTCN2020133493-appb-000423
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成2-氨基-4-氯-N-甲基苯甲酰胺Step A: Synthesis of 2-amino-4-chloro-N-methylbenzamide
Figure PCTCN2020133493-appb-000424
Figure PCTCN2020133493-appb-000424
取2-氨基-4-氯苯甲酸(1.71克，10毫摩尔)加入四氢呋喃(10毫升)中，再加入N,N'-羰基二咪唑(1.82克，11毫摩尔)，室温条件下反应1小时，1小时后，逐滴加入30％的甲胺甲醇溶液(2.24毫升，20毫摩尔)，反应12小时后，加入乙酸乙酯(50毫升)稀释反应液，转移到分液漏斗中，用饱和食盐水洗1次，乙酸乙酯溶液用无水硫酸钠干燥，浓缩后柱层析纯化分离(正己烷/乙酸乙酯＝1/1)，得到1.0克产物(收率：54％)。LCMS:RT＝3.03min,[M+H] +＝185.09。 Add 2-amino-4-chlorobenzoic acid (1.71 g, 10 mmol) to tetrahydrofuran (10 mL), then add N,N'-carbonyldiimidazole (1.82 g, 11 mmol), and react at room temperature for 1 After 1 hour, 30% methylamine methanol solution (2.24 ml, 20 mmol) was added dropwise, after 12 hours of reaction, ethyl acetate (50 ml) was added to dilute the reaction solution, and transferred to a separatory funnel. Wash with saturated brine once, dry the ethyl acetate solution with anhydrous sodium sulfate, concentrate and purify and separate by column chromatography (n-hexane/ethyl acetate=1/1) to obtain 1.0 g of product (yield: 54%). LCMS: RT = 3.03 min, [M+H] + =185.09.
步骤B：合成2-((5-氯-2-(甲基氨基甲酰基)苯基)氨基)-2-氧代乙酸甲酯Step B: Synthesis of methyl 2-((5-chloro-2-(methylcarbamoyl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000425
Figure PCTCN2020133493-appb-000425
将2-氨基-4-氯-N-甲基苯甲酰胺(1.0克，5.4毫摩尔)溶解于二氯甲烷(10.8毫升)中，加入吡啶(0.87毫升，10.8毫摩尔)，冰水浴条件下，逐滴加入草酰氯单甲酯(0.6毫升，6.5毫摩尔)，加入完成后逐渐升至室温反应一小时。Dissolve 2-amino-4-chloro-N-methylbenzamide (1.0 g, 5.4 mmol) in dichloromethane (10.8 mL), add pyridine (0.87 mL, 10.8 mmol), and under ice-water bath conditions , Oxalyl chloride monomethyl ester (0.6 ml, 6.5 mmol) was added dropwise, and after the addition was completed, it was gradually raised to room temperature and reacted for one hour.
反应结束后，加入二氯甲烷稀释反应液，用饱和氯化铵溶液洗一次，二氯甲烷溶液用无水硫酸钠干燥，浓缩后柱层析纯化分离(正己烷/乙酸乙酯＝1/1)，得到600毫克产物(收率：41％)。LCMS:RT＝3.33min,[M+H] +＝271.09。 After the reaction, dichloromethane was added to dilute the reaction solution, washed once with saturated ammonium chloride solution, the dichloromethane solution was dried over anhydrous sodium sulfate, concentrated, and purified and separated by column chromatography (n-hexane/ethyl acetate = 1/1 ) To obtain 600 mg of product (yield: 41%). LCMS: RT = 3.33 min, [M+H] + =271.09.
步骤B：合成2-((5-氯-2-(甲基氨基甲酰基)苯基)氨基)-2-氧乙酸Step B: Synthesis of 2-((5-chloro-2-(methylcarbamoyl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000426
Figure PCTCN2020133493-appb-000426
将2-((5-氯-2-(甲基氨基甲酰基)苯基)氨基)-2-氧代乙酸甲酯(250毫克，0.92毫摩尔)溶解于四氢呋喃/水的混合溶液中(12毫升/6毫升)中，加入氢氧化锂(39毫克，0.92毫摩尔)。Methyl 2-((5-chloro-2-(methylcarbamoyl)phenyl)amino)-2-oxoacetate (250 mg, 0.92 mmol) was dissolved in a mixed solution of tetrahydrofuran/water (12 ML/6 mL) was added lithium hydroxide (39 mg, 0.92 mmol).
反应结束后，加入二氯甲烷稀释反应液，水相用2N的盐酸溶液调Ph至2-3，二氯甲烷萃取三次，浓缩后得到56毫克白色固体，粗品直接投下一步反应。LCMS:RT＝3.00min,[M-H] -＝255.07。 After the completion of the reaction, dichloromethane was added to dilute the reaction solution, the aqueous phase was adjusted to Ph to 2-3 with 2N hydrochloric acid solution, and the dichloromethane extracted three times. After concentration, 56 mg of white solid was obtained. The crude product was directly used for the next reaction. LCMS: RT = 3.00 min, [MH] - = 255.07.
步骤C：合成(S)-4-(2-(2-(((5-氯-2-(甲基氨基甲酰基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(methylcarbamoyl)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000427
Figure PCTCN2020133493-appb-000427
将2-((5-氯-2-(甲基氨基甲酰基)苯基)氨基)-2-氧乙酸(56毫克，0.218毫摩尔)，(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(74毫克，0.218毫摩尔)，2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(125毫克，0.33毫摩尔)加入烧瓶中，加入N,N-二甲基甲酰胺2毫升将底物全部溶解，最后加入N，N-二异丙基乙胺(0.1毫升，0.65毫摩尔)，室温条件下反应2小时。The 2-((5-chloro-2-(methylcarbamoyl)phenyl)amino)-2-oxoacetic acid (56 mg, 0.218 mmol), (S)-4-(2-amino-3- Phenylpropionamido) tert-butyl benzoate (74 mg, 0.218 mmol), 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate Salt (125 mg, 0.33 mmol) was added to the flask, 2 mL of N,N-dimethylformamide was added to dissolve all the substrate, and finally N,N-diisopropylethylamine (0.1 mL, 0.65 mmol) ), react for 2 hours at room temperature.
反应结束后，向反应液中加入乙酸乙酯(50毫升)并转移到分液漏斗中，用饱和食盐水洗4次(50毫升×4)，将乙酸乙酯层转移到锥形瓶中并加入无水硫酸钠干燥，过滤浓缩后得到油状液体，柱层析(洗脱剂：正己烷/乙酸乙酯＝1/1)纯化后得到30毫克产物(S)-4-(2-(2-(((5-氯-2-(甲基氨基甲酰基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：22％)。LCMS:RT＝4.34min,[M+H] +＝579.15。 After the reaction, add ethyl acetate (50 ml) to the reaction solution and transfer to a separatory funnel, wash 4 times with saturated brine (50 ml × 4), transfer the ethyl acetate layer to an Erlenmeyer flask and add It was dried over anhydrous sodium sulfate, filtered and concentrated to obtain an oily liquid. After purification by column chromatography (eluent: n-hexane/ethyl acetate = 1/1), 30 mg of product (S)-4-(2-(2- (((5-Chloro-2-(methylcarbamoyl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)tert-butyl benzoate (Yield: 22% ). LCMS: RT = 4.34 min, [M+H] + = 579.15.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(甲基氨基甲酰基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(methylcarbamoyl)phenyl)amino)-2-oxoacetamido)-3-phenyl Propionamido) benzoic acid
Figure PCTCN2020133493-appb-000428
Figure PCTCN2020133493-appb-000428
将(S)-4-(2-(2-(((5-氯-2-(甲基氨基甲酰基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(30毫克，0.05毫摩尔)加入二氯甲烷(2.5毫升)中，再逐滴加入三氟乙酸(0.4毫升，5.4毫摩尔)，室温条件下反应四小时，反应结束后直接浓缩蒸干得到白色固体，白色固体用二氯甲烷洗三次，过滤后得到12.9毫克产物。LCMS:RT＝3.69min,[M-H] -＝521.15, 1H NMR(400MHz,DMSO)δ(ppm)12.79(s,1H),12.74(s,1H),10.45(s,1H),9.09(d,J＝6.4Hz,1H),8.08(d,J＝3.6Hz,1H),8.06(d,J＝2.4Hz,1H),7.92(d,J＝6.8Hz,2H),7.79(d,J＝6.8Hz,1H),7.72(d,J＝6.8Hz,2H),7.42-7.22(m,5H),7.29(t,J＝6.0Hz,1H),4.78-4.70(m,1H),3.25-3.15(m,2H),2.77(d,J＝4.0Hz,3H)。 (S)-4-(2-(2-(((5-chloro-2-(methylcarbamoyl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamide Add tert-butyl benzoate (30 mg, 0.05 mmol) to dichloromethane (2.5 mL), then add trifluoroacetic acid (0.4 mL, 5.4 mmol) dropwise, and react for 4 hours at room temperature. The reaction is complete Then it was concentrated and evaporated to dryness to obtain a white solid, which was washed three times with dichloromethane, and filtered to obtain 12.9 mg of product. LCMS: RT = 3.69 min, [MH] - = 521.15, 1 H NMR (400MHz, DMSO) δ (ppm ) 12.79 (s, 1H), 12.74 (s, 1H), 10.45 (s, 1H), 9.09 (d, J = 6.4 Hz, 1H), 8.08 (d, J = 3.6 Hz, 1H), 8.06 (d, J = 2.4Hz, 1H), 7.92 (d, J = 6.8 Hz, 2H), 7.79 (d, J = 6.8 Hz, 1H), 7.72 (d, J = 6.8 Hz, 2H), 7.42-7.22 (m, 5H), 7.29 (t, J=6.0 Hz, 1H), 4.78-4.70 (m, 1H), 3.25-3.15 (m, 2H), 2.77 (d, J=4.0 Hz, 3H).
实施例75Example 75
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenyl Propionamido) benzoic acid
Figure PCTCN2020133493-appb-000429
Figure PCTCN2020133493-appb-000429
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-(((叔丁氧羰基)氨基)-3.-苯基丙酰胺基)苯甲酸甲酯Step A: Synthesis of methyl (S)-4-(2-(((tert-butoxycarbonyl)amino)-3.-phenylpropionamido)benzoate
Figure PCTCN2020133493-appb-000430
Figure PCTCN2020133493-appb-000430
将(叔丁氧羰基)-L-苯丙氨酸(300毫克，0.75毫摩尔)和4-氨基苯甲酸甲酯(95毫克，0.62毫摩尔)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(358毫克,0.94毫摩尔)，N,N-二异丙基乙胺(162毫克，1.2毫摩尔)，在室温下搅拌18个小时。(Tert-Butoxycarbonyl)-L-phenylalanine (300 mg, 0.75 mmol) and methyl 4-aminobenzoate (95 mg, 0.62 mmol) were dissolved in N,N-dimethylformamide ( 5.0 ml) of the solution. Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (358 mg, 0.94 mmol) was added to the above solution, N, N -Diisopropylethylamine (162 mg, 1.2 mmol), stirred at room temperature for 18 hours.
将反应液缓慢滴加到饱和氯化铵溶液(50毫升)中。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(15毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到300毫克黄色油状(S)-4-(2-(((叔丁氧羰基)氨基)-3-苯基丙酰胺基)苯甲酸甲酯(收率：120％)。MS(ESI)M/Z：399.18[M+H] +。 The reaction solution was slowly added dropwise to saturated ammonium chloride solution (50 mL). The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (15 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtained 300 mg of methyl (S)-4-(2-(((tert-butoxycarbonyl)amino)-3-phenylpropionamido)benzoate as a yellow oil (yield: 120%). MS(ESI)M /Z: 399.18[M + H] +.
步骤B：合成(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸甲酯盐酸盐Step B: Synthesis of methyl (S)-4-(2-amino-3-phenylpropionamido)benzoate hydrochloride
Figure PCTCN2020133493-appb-000431
Figure PCTCN2020133493-appb-000431
将(S)-4-(2-(((叔丁氧羰基)氨基)-3-苯基丙酰胺基)苯甲酸甲酯(300毫克，0.75毫摩尔)溶于乙酸乙酯(4毫升)。随后，向上述溶液中加入盐酸乙酸乙酯溶液(2摩尔/升，4毫升,)。在室温下搅拌4小时。Methyl (S)-4-(2-(((tert-butoxycarbonyl)amino)-3-phenylpropionamido)benzoate (300 mg, 0.75 mmol) was dissolved in ethyl acetate (4 mL) Subsequently, a hydrochloric acid ethyl acetate solution (2 mol/L, 4 mL,) was added to the above solution, and the mixture was stirred at room temperature for 4 hours.
将反应液减压浓缩。得到150毫油状物(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸甲酯盐酸盐(收率：59.0％)。MS(ESI)M/Z：299.18[M+H] +。 The reaction solution was concentrated under reduced pressure. 150 milliliters of oily (S)-4-(2-amino-3-phenylpropionamido)methyl benzoate hydrochloride was obtained (yield: 59.0%). MS (ESI) M/Z: 299.18 [M+H] + .
步骤C：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯Step C: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-phenylpropionamido) methyl benzoate
Figure PCTCN2020133493-appb-000432
Figure PCTCN2020133493-appb-000432
将(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸甲酯盐酸盐(100毫克，0.33毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(98毫克，0.33毫摩尔)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50％质量分数的1-丙基磷酸酐N,N-二甲基甲酰胺溶液(523毫克,1.68毫摩尔)，N,N-二异丙基乙胺(129毫克，1.01毫摩尔)，在室温下搅拌4个小时。Combine (S)-4-(2-amino-3-phenylpropionamido) benzoic acid methyl ester hydrochloride (100 mg, 0.33 mmol) and 2-((5-chloro-2-(1H-tetra Azol-1-yl)phenyl)amino)-2-oxoacetic acid (98 mg, 0.33 mmol) was dissolved in a solution of N,N-dimethylformamide (5.0 mL). Subsequently, to the above solution was added 50% mass fraction of 1-propyl phosphoric anhydride N,N-dimethylformamide solution (523 mg, 1.68 mmol), N, N-diisopropylethylamine (129 mg , 1.01 mmol), stirred at room temperature for 4 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(25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：乙酸乙酯/正己烷＝1/2)。得到120毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯(收率：67.0％)。MS(ESI)M/Z：548.15[M+H] +。 Saturated ammonium chloride solution (20 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (25 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: ethyl acetate/n-hexane = 1/2). Obtain 120 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) Methyl-3-phenylpropionamido)benzoate (yield: 67.0%). MS (ESI) M/Z: 548.15 [M+H] + .
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000433
Figure PCTCN2020133493-appb-000433
将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯(50毫克，91微摩尔)溶于四氢呋喃/水(2/1，3毫升)的溶液中。随后，向上述溶液中加入氢氧化锂(8.3毫克,0.18毫摩尔)，在室温下搅拌3个小时。(S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene Methyl propionamido) benzoate (50 mg, 91 micromoles) was dissolved in a solution of tetrahydrofuran/water (2/1, 3 ml). Then, lithium hydroxide (8.3 mg, 0.18 ml) was added to the above solution. Mol) and stirred at room temperature for 3 hours.
向反应液中加入1摩尔/升盐酸溶液，调pH至3。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：甲醇/二氯甲烷＝1/10)。得到18毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：36.9％)。MS(ESI)M/Z：534.22[M+H] +。 Add 1 mol/L hydrochloric acid solution to the reaction solution to adjust the pH to 3. 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: methanol/dichloromethane = 1/10). Obtain 18 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-Phenylpropionamido)benzoic acid (yield: 36.9%). MS (ESI) M/Z: 534.22 [M+H] + .
实施例76Example 76
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(吡啶-4-基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(pyridine -4-yl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000434
Figure PCTCN2020133493-appb-000434
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(吡啶-4-基)丙酰胺基)苯甲酸甲酯Step A: Synthesis of methyl (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(pyridin-4-yl)propionamido)benzoate
Figure PCTCN2020133493-appb-000435
Figure PCTCN2020133493-appb-000435
将(S)-2-((叔丁氧羰基)氨基)-3-(吡啶-4-基)丙酸(400毫克，1.5毫摩尔)和4-氨基苯甲酸甲酯(206毫克，1.37毫摩尔)溶于N,N-二甲基甲酰胺(10毫升)的溶液中。随后，向上述溶液中加入1-丙基磷酸酐N,N-二甲基甲酰胺溶液(1.28克,4.1毫摩尔)，N,N-二异丙基乙胺(264毫克，2.06毫摩尔)，在室温下搅拌3个小时。(S)-2-((tert-butoxycarbonyl)amino)-3-(pyridin-4-yl)propionic acid (400 mg, 1.5 mmol) and methyl 4-aminobenzoate (206 mg, 1.37 mmol) Mol) was dissolved in a solution of N,N-dimethylformamide (10 mL). Subsequently, 1-propyl phosphoric anhydride N,N-dimethylformamide solution (1.28 g, 4.1 mmol) and N, N-diisopropylethylamine (264 mg, 2.06 mmol) were added to the above solution. , Stir at room temperature for 3 hours.
将反应液缓慢滴加到饱和氯化铵溶液(50毫升)中。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(15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＝1/2)得到220毫克黄色油状(S)-4-(2-((叔丁氧羰基)氨基)-3-(吡啶-4-基)丙酰胺基)苯甲酸甲酯(收率：40.0％)。MS(ESI)M/Z：400.04[M+H] +。 The reaction solution was slowly added dropwise to saturated ammonium chloride solution (50 mL). The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (15 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 = 1/2) to obtain 220 mg of (S)-4-(2-((tert-butoxycarbonyl)amino)-3 as a yellow oil Methyl (pyridin-4-yl)propionamido)benzoate (yield: 40.0%). MS (ESI) M/Z: 400.04 [M+H] + .
步骤B：合成(S)-4-(2-氨基-3-(吡啶-4-基)丙酰胺基)苯甲酸甲酯盐酸盐Step B: Synthesis of methyl (S)-4-(2-amino-3-(pyridin-4-yl)propionamido)benzoate hydrochloride
Figure PCTCN2020133493-appb-000436
Figure PCTCN2020133493-appb-000436
将(S)-4-(2-((叔丁氧羰基)氨基)-3-(吡啶-4-基)丙酰胺基)苯甲酸甲酯(220毫克，0.37毫摩尔)溶于乙酸乙酯(4毫升)。随后，向上述溶液中加入盐酸乙酸乙酯溶液(2摩尔/升，4毫升,)。在室温下搅拌4小时。Methyl (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(pyridin-4-yl)propionamido)benzoate (220 mg, 0.37 mmol) was dissolved in ethyl acetate (4 ml). Subsequently, a hydrochloric acid ethyl acetate solution (2 mol/L, 4 mL,) was added to the above solution. Stir at room temperature for 4 hours.
将反应液减压浓缩。得到190毫油状物(S)-4-(2-氨基-3-(吡啶-4-基)丙酰胺基)苯甲酸甲酯盐酸盐(收率：92.6％)。MS(ESI)M/Z：300.13[M+H] +。 The reaction solution was concentrated under reduced pressure. 190 milliliters of (S)-4-(2-amino-3-(pyridin-4-yl)propionamido)benzoic acid methyl ester hydrochloride (yield: 92.6%) was obtained. MS (ESI) M/Z: 300.13 [M+H] + .
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(吡啶-4-基)丙酰胺基)苯甲酸甲酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(Pyridin-4-yl)propionamido)methyl benzoate
Figure PCTCN2020133493-appb-000437
Figure PCTCN2020133493-appb-000437
将(S)-4-(2-氨基-3-(吡啶-4-基)丙酰胺基)苯甲酸甲酯盐酸盐(100毫克，0.66毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(196毫克，0.73毫摩尔)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50％质量分数的1-丙基磷酸酐N,N-二甲基甲酰胺溶液(834毫克,2.6毫摩尔)，N,N-二异丙基乙胺(337毫克，2.67毫摩尔)，在室温下搅拌18个小时。Combine (S)-4-(2-amino-3-(pyridin-4-yl)propionamido) benzoic acid methyl ester hydrochloride (100 mg, 0.66 mmol) and 2-((5-chloro-2 -(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (196 mg, 0.73 mmol) was dissolved in a solution of N,N-dimethylformamide (10.0 mL). Subsequently, 50% mass fraction of 1-propyl phosphoric anhydride N,N-dimethylformamide solution (834 mg, 2.6 mmol), N,N-diisopropylethylamine (337 mg , 2.67 mmol) and stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(50毫升)。混合液用乙酸乙酯(25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：乙酸乙酯/正己烷＝100/0)。得到50毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(吡啶-4-基)丙酰胺基)苯甲酸甲酯(收率：34.0％)。MS(ESI)M/Z：549.21[M+H] +。 Saturated ammonium chloride solution (50 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (25 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: ethyl acetate/n-hexane=100/0). Obtain 50 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Methyl 3-(pyridin-4-yl)propionamido)benzoate (yield: 34.0%). MS (ESI) M/Z: 549.21 [M+H] + .
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(吡啶-4-基)丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(Pyridin-4-yl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000438
Figure PCTCN2020133493-appb-000438
将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯(50毫克，91微摩尔)溶于四氢呋喃/水(2/1，3毫升)的溶液中。随后，向上述溶液中加入氢氧化锂(7.6毫克,0.18毫摩尔)，在室温下搅拌3个小时。(S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-benzene Methyl propionamido) benzoate (50 mg, 91 micromoles) was dissolved in a solution of tetrahydrofuran/water (2/1, 3 ml). Then, lithium hydroxide (7.6 mg, 0.18 ml) was added to the above solution. Mol) and stirred at room temperature for 3 hours.
向反应液中加入1摩尔/升盐酸溶液，调pH至3。有大量固体析出，过滤，用水洗涤。滤饼干燥得到15.9毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(吡啶-4-基)丙酰胺基)苯甲酸(收率：32.6％)。MS(ESI)M/Z：533.22[M-H] -。 1H NMR(400MHz,DMSO)δ12.78(s,1H),10.78(s,1H),10.68(s,1H),9.81(s,1H),9.27(d,J＝8.6Hz,1H),8.80(d,J＝6.1Hz,2H),7.92(d,J＝8.9Hz,4H),7.75(t,J＝8.5Hz,3H),7.62(dd,J＝8.6,2.3Hz,1H),4.90(td,J＝10.3,4.0Hz,1H),3.71(d,J＝89.9Hz,2H)。 Add 1 mol/L hydrochloric acid solution to the reaction solution to adjust the pH to 3. A large amount of solid precipitated, filtered and washed with water. The filter cake was dried to obtain 15.9 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-(pyridin-4-yl)propionamido)benzoic acid (yield: 32.6%). MS(ESI) M/Z: 533.22 [MH] - . 1 H NMR(400MHz,DMSO)δ12.78(s,1H), 10.78(s,1H), 10.68(s,1H), 9.81(s,1H), 9.27(d,J=8.6Hz,1H), 8.80(d,J=6.1Hz,2H),7.92(d,J=8.9Hz,4H),7.75(t,J=8.5Hz,3H), 7.62(dd,J=8.6,2.3Hz,1H), 4.90 (td, J = 10.3, 4.0 Hz, 1H), 3.71 (d, J = 89.9 Hz, 2H).
实施例77Example 77
合成(S)-4-(2-(2-(((5-氯-2-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000439
Figure PCTCN2020133493-appb-000439
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成2-((5-氯-2-氰基苯基)氨基)-2-氧乙酸甲酯Step A: Synthesis of methyl 2-((5-chloro-2-cyanophenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000440
Figure PCTCN2020133493-appb-000440
将2-氨基-4-氯苄腈(300毫克，1.97毫摩尔)和2-氯-2-氧代乙酸甲酯(361毫克，2.95毫摩尔)溶于二氯甲烷(10.0毫升)的溶液中。随后，向上述溶液中加入三乙胺(394毫克,3.94毫摩尔)，在室温下搅拌2个小时。Dissolve 2-amino-4-chlorobenzonitrile (300 mg, 1.97 mmol) and methyl 2-chloro-2-oxoacetate (361 mg, 2.95 mmol) in dichloromethane (10.0 mL) . Subsequently, triethylamine (394 mg, 3.94 mmol) was added to the above solution and stirred at room temperature for 2 hours.
将反应液缓慢滴加到饱和氯化铵溶液(20毫升)中。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(15毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到460毫克黄色油状2-((5-氯-2-氰基苯基)氨基)-2-氧乙酸甲酯(收率：98.0％)。MS(ESI)M/Z：239.01[M+H] +。 The reaction solution was slowly added dropwise to saturated ammonium chloride solution (20 mL). The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first saturated brine (15 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. 460 mg of methyl 2-((5-chloro-2-cyanophenyl)amino)-2-oxoacetate was obtained as a yellow oil (yield: 98.0%). MS (ESI) M/Z: 239.01 [M+H] + .
步骤B：合成2-((5-氯-2-氰基苯基)氨基)-2-氧乙酸Step B: Synthesis of 2-((5-chloro-2-cyanophenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000441
Figure PCTCN2020133493-appb-000441
将(2-((5-氯-2-氰基苯基)氨基)-2-氧乙酸甲酯(460毫克，1.93毫摩尔)溶于四氢呋喃/水(2/1，15毫升)的溶液中。随后，向上述溶液中加入氢氧化锂(161毫克,3.86毫摩尔)，在室温下搅拌1个小时。(2-((5-Chloro-2-cyanophenyl)amino)-2-oxoacetic acid methyl ester (460 mg, 1.93 mmol) was dissolved in a solution of tetrahydrofuran/water (2/1, 15 ml) Subsequently, lithium hydroxide (161 mg, 3.86 mmol) was added to the above solution and stirred at room temperature for 1 hour.
向反应液中加入1摩尔/升盐酸溶液，调pH至3。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：甲醇/二氯甲烷＝1/10)。得到250毫克白色固体2-((5-氯-2-氰基苯基)氨基)-2-氧乙酸(收率：57.7％)。MS(ESI)M/Z：223.20[M-H] -。 Add 1 mol/L hydrochloric acid solution to the reaction solution to adjust the pH to 3. 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: methanol/dichloromethane = 1/10). 250 mg of white solid 2-((5-chloro-2-cyanophenyl)amino)-2-oxoacetic acid was obtained (yield: 57.7%). MS(ESI) M/Z: 223.20 [MH] - .
步骤C：合成(S)-4-(2-(2-(((5-氯-2-氰基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-(((5-chloro-2-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzene Tert-butyl formate
Figure PCTCN2020133493-appb-000442
Figure PCTCN2020133493-appb-000442
将2-((5-氯-2-氰基苯基)氨基)-2-氧乙酸(101毫克，0.45毫摩尔)和(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯盐酸盐(200毫克，0.41毫摩尔)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50％质量分数的1-丙基磷酸酐N,N-二甲基甲酰胺溶液(513毫克,1.63毫摩尔)，N,N-二异丙基乙胺(159毫克，1.23毫摩尔)，在室温下搅拌5个小时。Combine 2-((5-chloro-2-cyanophenyl)amino)-2-oxoacetic acid (101 mg, 0.45 mmol) and (S)-4-(2-amino-3-phenylpropionamido ) Tert-butyl benzoate hydrochloride (200 mg, 0.41 mmol) was dissolved in a solution of N,N-dimethylformamide (10.0 mL). Subsequently, 50% mass fraction of 1-propyl phosphoric anhydride N,N-dimethylformamide solution (513 mg, 1.63 mmol), N,N-diisopropylethylamine (159 mg) was added to the above solution. , 1.23 mmol) and stirred at room temperature for 5 hours.
向反应液中加入饱和氯化铵溶液(50毫升)。混合液用乙酸乙酯(25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：乙酸乙酯/正己烷＝1/5)。得到100毫克白色固体(S)-4-(2-(2-(((5-氯-2-氰基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：44.0％)。MS(ESI)M/Z：547.20[M+H] +。 Saturated ammonium chloride solution (50 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (25 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: ethyl acetate/n-hexane=1/5). Obtain 100 mg of white solid (S)-4-(2-(2-(((5-chloro-2-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) Tert-butyl benzoate (yield: 44.0%). MS (ESI) M/Z: 547.20 [M+H] + .
步骤D：合成(S)-4-(2-(2-(((5-氯-2-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-(((5-chloro-2-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000443
Figure PCTCN2020133493-appb-000443
将(S)-4-(2-(2-(((5-氯-2-氰基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(80毫克，146微摩尔)溶于二氯甲烷(5毫升)的溶液中。随后，向上述溶液中加入三氟乙酸(1毫升)，在室温下搅拌3个小时。Add (S)-4-(2-(2-(((5-chloro-2-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) tert-butyl benzoate The ester (80 mg, 146 μmol) was dissolved in a solution of dichloromethane (5 mL). Then, trifluoroacetic acid (1 mL) was added to the above solution, and the mixture was stirred at room temperature for 3 hours.
将反应液减压浓缩。所得残余物用乙酸乙酯10(毫升)打浆，过滤。得到20毫克白色固体(S)-4-(2-(2-(((5-氯-2-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：27.8％)。MS(ESI)M/Z：489.10[M-H] -。 1H NMR(400MHz,DMSO)δ12.75(s,1H),10.87(s,1H),10.51(s,1H),9.20(d,J＝8.2Hz,1H),7.92(dd,J＝8.6,3.7Hz,3H),7.79(d,J＝1.9Hz,1H),7.71(d,J＝8.7Hz,2H),7.53(dd,J＝8.4,2.0Hz,1H),7.36–7.24(m,4H),7.20(t,J＝7.1Hz,1H),4.78(dd,J＝15.1,7.4Hz,1H),3.19(d,J＝7.2Hz,2H). The reaction solution was concentrated under reduced pressure. The resulting residue was slurried with ethyl acetate 10 (ml) and filtered. Obtain 20 mg of white solid (S)-4-(2-(2-(((5-chloro-2-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido ) Benzoic acid (yield: 27.8%). MS (ESI) M/Z: 489.10 [MH] - . 1 H NMR (400MHz, DMSO) δ 12.75 (s, 1H), 10.87 (s, 1H), 10.51 (s, 1H), 9.20 (d, J = 8.2 Hz, 1H), 7.92 (dd, J = 8.6, 3.7 Hz, 3H), 7.79 (d, J = 1.9 Hz, 1H), 7.71 (d, J = 8.7Hz, 2H), 7.53 (dd, J = 8.4, 2.0 Hz, 1H), 7.36-7.24 (m, 4H), 7.20 (t, J = 7.1 Hz, 1H), 4.78 (dd, J = 15.1, 7.4 Hz, 1H), 3.19 (d, J = 7.2 Hz, 2H).
实施例78Example 78
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-甲基丁酰胺基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-Methylbutyramido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000444
Figure PCTCN2020133493-appb-000444
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-(4-硝基苯基)丙酰氨基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-nitrophenyl)propionylamino)benzoic acid Tert-butyl ester
Figure PCTCN2020133493-appb-000445
Figure PCTCN2020133493-appb-000445
将(S)-2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-(4-硝基苯基)丙酸(1.72克，3.98毫摩尔)和4-氨基苯甲酸甲酯(700毫克，3.6毫摩尔)溶于N,N-二甲基甲酰胺(15.0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(2.07克,5.43毫摩尔)，N,N-二异丙基乙胺(936毫克，7.24毫摩尔)，在室温下搅拌18个小时。Combine (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-nitrophenyl)propionic acid (1.72 g, 3.98 mmol) and 4 -Methyl aminobenzoate (700 mg, 3.6 mmol) was dissolved in a solution of N,N-dimethylformamide (15.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.07 g, 5.43 mmol) was added to the above solution, N, N -Diisopropylethylamine (936 mg, 7.24 mmol), stirred at room temperature for 18 hours.
将反应液缓慢滴加到饱和氯化铵溶液(100毫升)中。混合液用乙酸乙酯(40毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(30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＝1/2)。得到1.8克黄色固体(S)-4-(2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-(4-硝基苯基)丙酰氨基)苯甲酸叔丁酯(收率：81.8％)。MS(ESI)M/Z：608.23[M+H] +。 The reaction solution was slowly added dropwise to saturated ammonium chloride solution (100 mL). The mixture was extracted with ethyl acetate (40 ml×3 times). Combine the organic phases. The organic phase was first washed with 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/n-hexane = 1 /2). Obtain 1.8 g of yellow solid (S)-4-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-nitrophenyl)propionylamino)benzene Tert-Butyl formate (yield: 81.8%). MS (ESI) M/Z: 608.23 [M+H] + .
步骤B：合成(S)-4-(2-氨基-3-(4-硝基苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of tert-butyl (S)-4-(2-amino-3-(4-nitrophenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000446
Figure PCTCN2020133493-appb-000446
将(S)-4-(2-((((9H-芴-9-基)甲氧基)羰基)氨基)-3-(4-硝基苯基)丙酰氨基)苯甲酸叔丁酯(1.8克，2.9毫摩尔)溶于二氯甲烷(5毫升)。随后，向上述溶液中加入4-甲基哌啶(1毫升,)。在室温下搅拌18小时。(S)-4-(2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-nitrophenyl)propionylamino)tert-butyl benzoate (1.8 g, 2.9 mmol) was dissolved in dichloromethane (5 mL). Subsequently, 4-methylpiperidine (1 mL,) was added to the above solution. Stir at room temperature for 18 hours.
将反应液缓慢滴加到饱和氯化铵溶液(20毫升)中。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(15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1.0克白色固体(S)-4-(2-氨基-3-(4-硝基苯基)丙酰胺基)苯甲酸叔丁酯(收率：87.0％)。MS(ESI)M/Z：386.20[M+H] +。 The reaction solution was slowly added dropwise to saturated ammonium chloride solution (20 mL). The mixture was extracted with ethyl acetate (20 ml×3 times). Combine the organic phases. The organic phase was first washed with saturated brine (15 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). 1.0 g of tert-butyl (S)-4-(2-amino-3-(4-nitrophenyl)propionamido)benzoate was obtained as a white solid (yield: 87.0%). MS (ESI) M/Z: 386.20 [M+H] + .
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-硝基苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-Nitrophenyl) propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000447
Figure PCTCN2020133493-appb-000447
将(S)-4-(2-氨基-3-(4-硝基苯基)丙酰胺基)苯甲酸叔丁酯(1.0克，2.6毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(764毫克，2.8毫摩尔)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50％质量分数的1-丙基磷酸酐N,N-二甲基甲酰胺溶液(3.2克,5.2毫摩尔)，N,N-二异丙基乙胺(1.1克，7.6毫摩尔)，在室温下搅拌18个小时。Combine (S)-4-(2-amino-3-(4-nitrophenyl) propionamido) tert-butyl benzoate (1.0 g, 2.6 mmol) and 2-((5-chloro-2- (1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (764 mg, 2.8 mmol) was dissolved in a solution of N,N-dimethylformamide (10.0 mL). Subsequently, to the above solution was added 50% mass fraction of 1-propyl phosphoric anhydride N,N-dimethylformamide solution (3.2 g, 5.2 mmol), N, N-diisopropylethylamine (1.1 g , 7.6 mmol) and stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(40毫升)。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×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400毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-硝基苯基)丙酰胺基)苯甲酸叔丁酯(收率：25.0％)。MS(ESI)M/Z：635.21[M+H] +。 Saturated ammonium chloride solution (40 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (30 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: ethyl acetate/n-hexane=1/1). Obtain 400 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-nitrophenyl)propionamido)benzoate (yield: 25.0%). MS (ESI) M/Z: 635.21 [M+H] + .
步骤D：合成(S)-4-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido) propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000448
Figure PCTCN2020133493-appb-000448
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-硝基苯基)丙酰胺基)苯甲酸叔丁酯(400毫克，1.57毫摩尔)溶于乙酸乙酯(20毫升)的溶液中。随后，向上述溶液中加入10％质量分数的湿Pd/C(80毫克)，在氢气球的条件下室温搅拌18个小时。(S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -Nitrophenyl) propionamido) tert-butyl benzoate (400 mg, 1.57 mmol) was dissolved in a solution of ethyl acetate (20 mL). Subsequently, 10% mass fraction of wet Pd/C (80 mg) was added to the above solution, and the mixture was stirred at room temperature for 18 hours under the condition of a hydrogen balloon.
将反应液过滤，滤饼用乙酸乙酯(50毫升)洗涤，滤液减压浓缩。得到300毫克黄色油状物(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：78.0％)。MS(ESI)M/Z：605.20[M+H] +。 The reaction solution was filtered, the filter cake was washed with ethyl acetate (50 mL), and the filtrate was concentrated under reduced pressure. Obtain 300 mg of yellow oil (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-phenylpropionamido)benzoic acid (yield: 78.0%). MS (ESI) M/Z: 605.20 [M+H] + .
步骤E：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-甲基丁二胺基)苯基)丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-Methylbutanediamino)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000449
Figure PCTCN2020133493-appb-000449
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(100毫克，0.49毫摩尔)和吡啶(78毫克，0.99毫摩尔)溶于二氯甲烷(10.0毫升)的溶液中。随后，向上述溶液中加入3-甲基丁酰氯(89毫克,0.74毫摩尔)，在室温下搅拌5个小时。(S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenyl Propionamido)benzoic acid (100 mg, 0.49 mmol) and pyridine (78 mg, 0.99 mmol) were dissolved in dichloromethane (10.0 mL). Subsequently, 3-methylbutyryl chloride (89 mg, 0.74 mmol) was added to the above solution, and stirred at room temperature for 5 hours.
向反应液中加入饱和碳酸氢钠溶液(15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(20毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到50毫克棕色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-甲基丁二胺基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：44.0％)。MS(ESI)M/Z：689.33[M+H] +。 Saturated sodium bicarbonate solution (15 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 (20 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 50 mg of brown solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(3-methylbutanediamino)phenyl)propionamido)benzoate (yield: 44.0%). MS (ESI) M/Z: 689.33 [M+H] + .
步骤F：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-甲基丁酰胺基)苯基)丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-Methylbutyramido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000450
Figure PCTCN2020133493-appb-000450
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-甲基丁二胺基)苯基)丙酰胺基)苯甲酸叔丁酯(50毫克，72微摩尔)溶于二氯甲烷(5毫升)的溶液中。随后，向上述溶液中加入三氟乙酸(1毫升)，在室温下搅拌4个小时。(S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-Methylbutanediamino)phenyl)propionamido)tert-butyl benzoate (50 mg, 72 micromole) was dissolved in dichloromethane (5 mL). Subsequently, trifluoroacetic acid (1 ml) was added to the above solution and stirred at room temperature for 4 hours.
将反应液减压浓缩。所得残余物用甲醇(3毫升)打浆，过滤，得到18.9毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-甲基丁酰胺基)苯基)丙酰胺基)苯甲酸(收率：41.0％)。MS(ESI)M/Z：633.20[M+H] +。 1H NMR(400MHz,DMSO)δ10.71(s,1H),10.45(s,1H),9.87–9.69(m,2H),8.91(d,J＝7.9Hz,1H),7.97(d,J＝2.2Hz,1H),7.90(d,J＝8.7Hz,2H),7.76(d,J＝8.6Hz,1H),7.68(d,J＝8.7Hz,2H),7.60(dd,J＝8.7,2.3Hz,1H),7.48(d,J＝8.5Hz,2H),7.17(d,J＝8.5Hz,2H),4.67(dd,J＝14.8,7.2Hz,1H),3.08(d,J＝7.0Hz,2H),2.18–1.98(m,3H),0.87(dd,J＝28.4,7.6Hz,6H). The reaction solution was concentrated under reduced pressure. The obtained residue was slurried with methanol (3 ml) and filtered to obtain 18.9 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(3-methylbutamido)phenyl)propionamido)benzoic acid (yield: 41.0%). MS (ESI) M/Z: 633.20 [M+H] + . 1 H NMR (400MHz, DMSO) δ 10.71 (s, 1H), 10.45 (s, 1H), 9.87-9.69 (m, 2H), 8.91 (d, J = 7.9 Hz, 1H), 7.97 (d, J ＝2.2Hz,1H),7.90(d,J＝8.7Hz,2H), 7.76(d,J＝8.6Hz,1H), 7.68(d,J＝8.7Hz,2H), 7.60(dd,J＝8.7 ,2.3Hz,1H),7.48(d,J=8.5Hz,2H),7.17(d,J=8.5Hz,2H), 4.67(dd,J=14.8,7.2Hz,1H),3.08(d,J =7.0Hz,2H), 2.18–1.98(m,3H), 0.87(dd,J=28.4,7.6Hz,6H).
实施例79Example 79
合成(S)-4-(2-(2-(((3-氯-2-氟苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((3-chloro-2-fluorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000451
Figure PCTCN2020133493-appb-000451
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成2-((3-氯-2-氟苯基)氨基)-2-氧乙酸甲酯Step A: Synthesis of methyl 2-((3-chloro-2-fluorophenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000452
Figure PCTCN2020133493-appb-000452
将3-氯-2-氟苯胺(300毫克，2毫摩尔)溶于二氯甲烷(10.0毫升)的溶液中。随后，向上述溶液中加入2-氯-2-氧代乙酸甲酯(376毫克，3.0毫摩尔)和三乙胺(413毫克，4.1毫摩尔)。在室温下搅拌2小时。3-Chloro-2-fluoroaniline (300 mg, 2 mmol) was dissolved in a solution of dichloromethane (10.0 mL). Subsequently, methyl 2-chloro-2-oxoacetate (376 mg, 3.0 mmol) and triethylamine (413 mg, 4.1 mmol) were added to the above solution. Stir at room temperature for 2 hours.
向反应液中加入饱和碳酸氢钠溶液(15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(20毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到280毫克棕色油状物2-((3-氯-2-氟苯基)氨基)-2-氧乙酸甲酯(收率：60.0％)。Saturated sodium bicarbonate solution (15 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 (20 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. 280 mg of methyl 2-((3-chloro-2-fluorophenyl)amino)-2-oxoacetate was obtained as a brown oil (yield: 60.0%).
步骤B：合成2-((3-氯-2-氟苯基)氨基)-2-氧乙酸Step B: Synthesis of 2-((3-chloro-2-fluorophenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000453
Figure PCTCN2020133493-appb-000453
将2-((3-氯-2-氟苯基)氨基)-2-氧乙酸甲酯(280毫克，1.2毫摩尔)溶于四氢呋喃/水(2/1，9毫升)的溶液中。随后，向上述溶液中加入氢氧化锂(11毫克，1.3毫摩尔)。在室温下搅拌0.5个小时。Methyl 2-((3-chloro-2-fluorophenyl)amino)-2-oxoacetate (280 mg, 1.2 mmol) was dissolved in a solution of tetrahydrofuran/water (2/1, 9 mL). Subsequently, lithium hydroxide (11 mg, 1.3 mmol) was added to the above solution. Stir at room temperature for 0.5 hour.
向反应液中加入饱和氯化铵溶液(15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。得到180毫克白色固体2-((3-氯-2-氟苯基)氨基)-2-氧乙酸(收率：66.0％)。MS(ESI)M/Z：216.00[M-H] -。 Saturated ammonium chloride solution (15 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. 180 mg of white solid 2-((3-chloro-2-fluorophenyl)amino)-2-oxoacetic acid was obtained (yield: 66.0%). MS (ESI) M/Z: 216.00 [MH] - .
步骤C：合成(S)-4-(2-(2-(((3-氯-2-氟苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-(((3-chloro-2-fluorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid Tert-butyl ester
Figure PCTCN2020133493-appb-000454
Figure PCTCN2020133493-appb-000454
将2-((3-氯-2-氟苯基)氨基)-2-氧乙酸(70毫克，0.32毫摩尔)和(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(187毫克，0.38毫摩尔)溶于N,N-二甲基甲酰胺(10.0毫升)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(183毫克，0.48毫摩尔)，N,N-二异丙基乙胺(83毫克，0.64毫摩尔)，在室温下搅拌18个小时。Combine 2-((3-chloro-2-fluorophenyl)amino)-2-oxoacetic acid (70 mg, 0.32 mmol) and (S)-4-(2-amino-3-phenylpropionamido) Tert-butyl benzoate (187 mg, 0.38 mmol) was dissolved in N,N-dimethylformamide (10.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (183 mg, 0.48 mmol) was added to the above solution, N,N -Diisopropylethylamine (83 mg, 0.64 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：乙酸乙酯/正己烷＝1/1)。得到40毫克无色油状物(S)-4-(2-(2-(((3-氯-2-氟苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：23.0％)。MS(ESI)M/Z：540.25[M+H] +。 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: ethyl acetate/n-hexane=1/1). Obtain 40 mg of colorless oil (S)-4-(2-(2-(((3-chloro-2-fluorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido ) Tert-Butyl benzoate (yield: 23.0%). MS (ESI) M/Z: 540.25 [M+H] + .
步骤D：合成(S)-4-(2-(2-(((3-氯-2-氟苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-(((3-chloro-2-fluorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzene Formic acid
Figure PCTCN2020133493-appb-000455
Figure PCTCN2020133493-appb-000455
将(S)-4-(2-(2-(((3-氯-2-氟苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(40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。Add (S)-4-(2-(2-(((3-chloro-2-fluorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) tert-butyl benzoate (40 mg, 0.07 mmol) was dissolved in dichloromethane (4.0 mL). Then, 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。用甲醇(3毫升)打浆，过滤得27.4毫克白色固体(S)-4-(2-(2-(((3-氯-2-氟苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：76.4％)。MS(ESI)M/Z：482.10[M-H] -。 1H NMR(400MHz,DMSO)δ12.75(s,1H),10.49(d,J＝10.3Hz,2H),9.11(d,J＝8.2Hz,1H),7.92(d,J＝8.6Hz,2H),7.71(d,J＝8.6Hz,2H),7.55(t,J＝7.2Hz,1H),7.47(t,J＝7.0Hz,1H),7.38–7.16(m,6H),4.77(dd,J＝15.2,7.4Hz,1H),3.18(d,J＝7.1Hz,2H)。 The reaction solution was concentrated under reduced pressure. Beat with methanol (3 ml) and filter to obtain 27.4 mg of white solid (S)-4-(2-(2-(((3-chloro-2-fluorophenyl)amino)-2-oxoacetamido) -3-phenylpropionamido)benzoic acid (yield: 76.4%). MS (ESI) M/Z: 482.10 [MH] - . 1 H NMR (400MHz, DMSO) δ 12.75 (s, 1H), 10.49 (d, J = 10.3 Hz, 2H), 9.11 (d, J = 8.2 Hz, 1H), 7.92 (d, J = 8.6 Hz, 2H), 7.71 (d, J = 8.6 Hz, 2H), 7.55 ( t,J=7.2Hz,1H),7.47(t,J=7.0Hz,1H),7.38–7.16(m,6H),4.77(dd,J＝15.2,7.4Hz,1H),3.18(d,J =7.1Hz, 2H).
实施例80Example 80
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基哌嗪-1-羧酰胺基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-methylpiperazine-1-carboxamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000456
Figure PCTCN2020133493-appb-000456
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((phenoxycarbonyl)amino)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000457
Figure PCTCN2020133493-appb-000457
将(S)-4-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯(30毫克，49微摩尔)溶于四氢呋喃(3毫升)溶液中。随后，向上述溶液中加入氯甲酸苯酯(15.5毫克，99微摩尔)。在室温下搅拌2小时。(S)-4-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxy Acetamido) propionamido) tert-butyl benzoate (30 mg, 49 micromoles) was dissolved in a solution of tetrahydrofuran (3 ml). Subsequently, phenyl chloroformate (15.5 mg, 99 μmol) was added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。得到37毫克固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：103％)。MS(ESI)M/Z：725.22[M+H] +。 The reaction solution was concentrated under reduced pressure. Obtain 37 mg of solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((phenoxycarbonyl)amino)phenyl)propionamido)tert-butyl benzoate (yield: 103%). MS (ESI) M/Z: 725.22 [M+H] + .
步骤B：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-哌嗪-1-甲酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-piperazine-1-carboxamido)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000458
Figure PCTCN2020133493-appb-000458
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)苯甲酸叔丁酯(37毫克，0.51毫摩尔)和1-甲基哌嗪(10.2毫克，0.10毫摩尔)溶于乙腈(5.0毫升)的溶液中。在50摄氏度下搅拌3个小时。(S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((Phenoxycarbonyl)amino)phenyl)propionamido)tert-butyl benzoate (37 mg, 0.51 mmol) and 1-methylpiperazine (10.2 mg, 0.10 mmol) dissolved in acetonitrile (5.0 mL) ) In the solution. Stir at 50 degrees Celsius for 3 hours.
将反应液减压浓缩。所得残余物用甲醇(3毫升)打浆，过滤。得到31毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-哌嗪-1-甲酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：96.0％)。MS(ESI)M/Z：731.31[M+H] +。 The reaction solution was concentrated under reduced pressure. The resulting residue was slurried with methanol (3 mL) and filtered. Obtain 31 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(4-piperazine-1-carboxamido)phenyl)propionamido)benzoate (yield: 96.0%). MS (ESI) M/Z: 731.31 [M+H] + .
步骤C：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基哌嗪-1-羧酰胺基)苯基)丙酰胺基)苯甲酸Step C: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-methylpiperazine-1-carboxamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000459
Figure PCTCN2020133493-appb-000459
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-哌嗪-1-甲酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(31毫克，0.028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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-Piperazine-1-carboxamido)phenyl)propionamido)tert-butyl benzoate (31 mg, 0.028 mmol) was dissolved in methylene chloride (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。所得残余物用乙酸乙酯(3毫升)打浆，过滤，得到22.5毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基哌嗪-1-羧酰胺基)苯基)丙酰胺基)苯甲酸(收率：76.0％)。MS(ESI)M/Z：675.21[M+H] +。 1H NMR(400MHz,DMSO)δ10.96(s,1H),10.68(d,J＝30.2Hz,2H),9.83(s,1H),8.87(d,J＝8.3Hz,1H),8.77(s,1H),7.96(d,J＝2.3Hz,1H),7.90(d,J＝8.7Hz,2H),7.73(dd,J＝20.4,8.7Hz,3H),7.60(dd,J＝8.7,2.3Hz,1H),7.36(d,J＝8.5Hz,2H),7.15(d,J＝8.5Hz,2H),4.69(dd,J＝14.2,7.9Hz,1H),4.20(s,2H),3.08(d,J＝6.3Hz,3H),2.96(s,3H),2.69(d,J＝23.7Hz,3H)。 The reaction solution was concentrated under reduced pressure. The obtained residue was slurried with ethyl acetate (3 ml) and filtered to obtain 22.5 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl )Phenyl)amino)-2-oxoacetamido)-3-(4-(4-methylpiperazine-1-carboxamido)phenyl)propionamido)benzoic acid (yield: 76.0% ). MS (ESI) M/Z: 675.21 [M+H] + . 1 H NMR (400MHz, DMSO) δ 10.96 (s, 1H), 10.68 (d, J = 30.2 Hz, 2H), 9.83 (s ,1H),8.87(d,J=8.3Hz,1H),8.77(s,1H),7.96(d,J=2.3Hz,1H),7.90(d,J=8.7Hz,2H),7.73(dd ,J=20.4,8.7Hz,3H), 7.60(dd,J=8.7,2.3Hz,1H), 7.36(d,J=8.5Hz,2H), 7.15(d,J=8.5Hz,2H), 4.69 (dd,J=14.2,7.9Hz,1H), 4.20(s,2H), 3.08(d,J=6.3Hz,3H), 2.96(s,3H), 2.69(d,J=23.7Hz,3H) .
实施例81Example 81
合成(S)-4-(3-(4-苯甲酰胺基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸Synthesis of (S)-4-(3-(4-benzamidophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000460
Figure PCTCN2020133493-appb-000460
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(3-(4-苯甲酰胺基苯基)-2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(3-(4-benzamidophenyl)-2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000461
Figure PCTCN2020133493-appb-000461
将(S)-4-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯(30毫克，49微摩尔)溶于二氯甲烷(3毫升)溶液中。随后，向上述溶液中加入苯甲酰氯(8.3毫克，59微摩尔)和吡啶(5毫克，59微摩尔)。在室温下搅拌2小时。(S)-4-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxy Acetamido) propionamido) tert-butyl benzoate (30 mg, 49 micromoles) was dissolved in a solution of dichloromethane (3 mL). Subsequently, benzoyl chloride (8.3 mg, 59 micromoles) and pyridine (5 mg, 59 micromoles) were added to the above solution. Stir at room temperature for 2 hours.
将反应液减压浓缩。得到30毫克固体(S)-4-(3-(4-苯甲酰胺基苯基)-2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯(收率：121％)。MS(ESI)M/Z：709.22[M+H] +。 The reaction solution was concentrated under reduced pressure. Obtain 30 mg of solid (S)-4-(3-(4-benzamidophenyl)-2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)propionamido)tert-butyl benzoate (Yield: 121%). MS (ESI) M/Z: 709.22 [M+H] + .
步骤B：合成(S)-4-(3-(4-苯甲酰胺基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸Step B: Synthesis of (S)-4-(3-(4-benzamidophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) Amino)-2-oxoacetamido)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000462
Figure PCTCN2020133493-appb-000462
将(S)-4-(3-(4-苯甲酰胺基苯基)-2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯(30毫克，0.042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)-4-(3-(4-benzamidophenyl)-2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino) -2-oxoacetamido) propionamido) tert-butyl benzoate (30 mg, 0.042 mmol) was dissolved in dichloromethane (4.0 mL). Then, 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。所得残余物用甲醇(3毫升)打浆，过滤，得到18毫克白色固体(S)-4-(3-(4-苯甲酰胺基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸(收率：56.0％)。MS(ESI)M/Z：653.16[M+H] +。 1H NMR(400MHz,DMSO)δ12.74(s,1H),10.71(s,1H),10.48(s,1H),10.19(s,1H),9.82(s,1H),8.95(d,J＝8.1Hz,1H),8.01–7.87(m,4H),7.76(d,J＝8.6Hz,1H),7.69(t,J＝8.8Hz,3H),7.63–7.48(m,3H),7.25(d,J＝8.5Hz,2H),4.71(dd,J＝14.8,7.1Hz,1H),3.13(d,J＝7.0Hz,2H). The reaction solution was concentrated under reduced pressure. The resulting residue was slurried with methanol (3 mL) and filtered to obtain 18 mg of white solid (S)-4-(3-(4-benzamidophenyl)-2-(2-((5-chloro-2 -(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)benzoic acid (yield: 56.0%). MS (ESI) M/Z: 653.16 [M+H] + . 1 H NMR (400MHz, DMSO) δ 12.74 (s, 1H), 10.71 (s, 1H), 10.48 (s, 1H), 10.19 (s, 1H), 9.82 (s, 1H), 8.95 (d, J =8.1Hz,1H), 8.01–7.87(m,4H), 7.76(d,J=8.6Hz,1H), 7.69(t,J=8.8Hz,3H), 7.63–7.48(m,3H), 7.25 (d,J=8.5Hz,2H), 4.71(dd,J=14.8,7.1Hz,1H), 3.13(d,J=7.0Hz,2H).
实施例82Example 82
合成(S)-4-(2-(2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(3,3-difluoroazetidine-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000463
Figure PCTCN2020133493-appb-000463
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成1-(4-氯-2-硝基苯基)-3,3-二氟氮杂环丁烷Step A: Synthesis of 1-(4-chloro-2-nitrophenyl)-3,3-difluoroazetidine
Figure PCTCN2020133493-appb-000464
Figure PCTCN2020133493-appb-000464
将4-氯-1-氟-2-硝基苯(500毫克，2.8毫摩尔)和3,3-二氟氮杂环丁烷(318毫克，3.4毫摩尔)溶于N,N-二甲基甲酰胺(10毫升)溶液中。随后，向上述溶液中加入碳酸钾(787毫克，5.8毫摩尔)。在50摄氏度下搅拌4小时。Dissolve 4-chloro-1-fluoro-2-nitrobenzene (500 mg, 2.8 mmol) and 3,3-difluoroazetidine (318 mg, 3.4 mmol) in N,N-dimethyl Methyl formamide (10 ml) solution. Subsequently, potassium carbonate (787 mg, 5.8 mmol) was added to the above solution. Stir at 50 degrees Celsius for 4 hours.
向反应液中加入饱和氯化铵溶液(50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。得到650毫克无色油状物1-(4-氯-2-硝基苯基)-3,3-二氟氮杂环丁烷(收率：91.2％)。MS(ESI)M/Z：249.02[M+H] +。 Saturated ammonium chloride solution (50 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. 650 mg of colorless oily 1-(4-chloro-2-nitrophenyl)-3,3-difluoroazetidine was obtained (yield: 91.2%). MS (ESI) M/Z: 249.02 [M+H] + .
步骤B：合成5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯胺Step B: Synthesis of 5-chloro-2-(3,3-difluoroazetidine-1-yl)aniline
Figure PCTCN2020133493-appb-000465
Figure PCTCN2020133493-appb-000465
将1-(4-氯-2-硝基苯基)-3,3-二氟氮杂环丁烷(650毫克，2.6毫摩尔)溶于乙酸乙酯(15毫升)中。随后，向上述溶液中加入二水合氯化亚锡(5.6毫克，26毫摩尔)。在室温下搅拌18小时。1-(4-Chloro-2-nitrophenyl)-3,3-difluoroazetidine (650 mg, 2.6 mmol) was dissolved in ethyl acetate (15 mL). Subsequently, stannous chloride dihydrate (5.6 mg, 26 mmol) was added to the above solution. Stir at room temperature for 18 hours.
向反应液中加入饱和碳酸氢钠溶液(100毫升)。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×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到600毫克无色油状物5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯胺(收率：105％)。MS(ESI)M/Z：219.10[M+H] +。 Saturated sodium bicarbonate solution (100 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (30 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. 600 mg of colorless oily 5-chloro-2-(3,3-difluoroazetidin-1-yl)aniline was obtained (yield: 105%). MS (ESI) M/Z: 219.10 [M+H] + .
步骤C：合成2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-(3,3-difluoroazetidin-1-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000466
Figure PCTCN2020133493-appb-000466
将5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯胺(600毫克，2.7毫摩尔)溶于二氯甲烷(10.0毫升)的溶液中。随后，向上述溶液中加入2-氯-2-氧代乙酸甲酯(403毫克，3.2毫摩尔)和吡啶(434毫克，5.5毫摩尔)。在室温下搅拌2小时。5-Chloro-2-(3,3-difluoroazetidin-1-yl)aniline (600 mg, 2.7 mmol) was dissolved in a solution of dichloromethane (10.0 mL). Subsequently, methyl 2-chloro-2-oxoacetate (403 mg, 3.2 mmol) and pyridine (434 mg, 5.5 mmol) were added to the above solution. Stir at room temperature for 2 hours.
向反应液中加入饱和碳酸氢钠溶液(15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(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：乙酸乙酯/正己烷＝1/1)，得到320毫克无色油状物2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酸甲酯(收率：38.5％)。MS(ESI)M/Z：305.10[M+H] +。 Saturated sodium bicarbonate solution (15 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 (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: ethyl acetate/n-hexane=1/1) to obtain 320 mg of colorless oily 2-((5-chloro-2-(3,3-difluoro Azetidin-1-yl)phenyl)amino)-2-oxoacetate methyl ester (yield: 38.5%). MS (ESI) M/Z: 305.10 [M+H] + .
步骤D：合成2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酸Step D: Synthesis of 2-((5-chloro-2-(3,3-difluoroazetidin-1-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000467
Figure PCTCN2020133493-appb-000467
将2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酸甲酯(320毫克，1毫摩尔)溶于四氢呋喃/水(2/1，15毫升)的溶液中。随后，向上述溶液中加入氢氧化锂(88毫克，2.1毫摩尔)。在室温下搅拌3个小时。Dissolve methyl 2-((5-chloro-2-(3,3-difluoroazetidin-1-yl)phenyl)amino)-2-oxoacetate (320 mg, 1 mmol) In a solution of tetrahydrofuran/water (2/1, 15 ml). Subsequently, lithium hydroxide (88 mg, 2.1 mmol) was added to the above solution. Stir at room temperature for 3 hours.
向反应液中加入1M盐酸，调pH值至3。混合液用乙酸乙酯(25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21毫克白色固体2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酸(收率：68.7％)。MS(ESI)M/Z：289.10[M-H] -。 Add 1M hydrochloric acid to the reaction solution and adjust the pH to 3. The mixture was extracted with ethyl acetate (25 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 21 mg of white solid 2-((5-chloro-2-(3,3-difluoroazetidin-1-yl)phenyl)amino)-2-oxoacetic acid (yield: 68.7%) . MS (ESI) M/Z: 289.10 [MH] - .
步骤E：合成(S)-4-(2-(2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(3,3-difluoroazetidin-1-yl)phenyl)amino)-2-oxo (Acetamido)-3-phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000468
Figure PCTCN2020133493-appb-000468
将合成2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酸(100毫克，0.34毫摩尔)和(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(106毫克，0.31毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(178毫克，0.47毫摩尔)，N,N-二异丙基乙胺(80毫克，0.62毫摩尔)，在室温下搅拌18个小时。The synthesis of 2-((5-chloro-2-(3,3-difluoroazetidine-1-yl)phenyl)amino)-2-oxoacetic acid (100 mg, 0.34 mmol) and ( S) tert-butyl-4-(2-amino-3-phenylpropionamido)benzoate (106 mg, 0.31 mmol) was dissolved in N,N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (178 mg, 0.47 mmol) was added to the above solution, N, N -Diisopropylethylamine (80 mg, 0.62 mmol), stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(25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(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/正己烷＝1/1)。得到38毫克无色油状物(S)-4-(2-(2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：20.0％)。MS(ESI)M/Z：613.20[M+H] +。 Saturated ammonium chloride solution (25 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 (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/n-hexane=1/1). Obtained 38 mg of colorless oil (S)-4-(2-(2-((5-chloro-2-(3,3-difluoroazetidine-1-yl)phenyl)amino)- 2-oxoacetamido)-3-phenylpropionamido)tert-butyl benzoate (yield: 20.0%). MS (ESI) M/Z: 613.20 [M+H] + .
步骤F：合成(S)-4-(2-(2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-((5-chloro-2-(3,3-difluoroazetidin-1-yl)phenyl)amino)-2-oxo (Acetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000469
Figure PCTCN2020133493-appb-000469
将(S)-4-(2-(2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(38毫克，0.06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)-4-(2-(2-((5-chloro-2-(3,3-difluoroazetidin-1-yl)phenyl)amino)-2-oxoacetamide (Yl)-3-phenylpropionamido) tert-butyl benzoate (38 mg, 0.06 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。用甲醇(3毫升)打浆，过滤得12.2毫克白色固体(S)-4-(2-(2-((5-氯-2-(3,3-二氟氮杂环丁烷-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：35.3％)。MS(ESI)M/Z：557.10[M+H] +。 The reaction solution was concentrated under reduced pressure. Beat with methanol (3 ml) and filter to obtain 12.2 mg of white solid (S)-4-(2-(2-((5-chloro-2-(3,3-difluoroazetidine-1-yl) )Phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid (yield: 35.3%). MS (ESI) M/Z: 557.10 [M+H] + .
实施例83Example 83
合成(S)-4-(2-(2-(((3-氯-5-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((3-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000470
Figure PCTCN2020133493-appb-000470
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成2-((3-氯-5-氰基苯基)氨基)-2-氧乙酸甲酯Step A: Synthesis of methyl 2-((3-chloro-5-cyanophenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000471
Figure PCTCN2020133493-appb-000471
将3-氨基-5-氯苄腈(1.0克，6.5毫摩尔)溶于二氯甲烷(10.0毫升)的溶液中。随后，向上述溶液中加入2-氯-2-氧代乙酸甲酯(1.2克，9.6毫摩尔)和吡啶(868毫克，11毫摩尔)。在室温下搅拌2小时。3-Amino-5-chlorobenzonitrile (1.0 g, 6.5 mmol) was dissolved in a solution of dichloromethane (10.0 mL). Subsequently, methyl 2-chloro-2-oxoacetate (1.2 g, 9.6 mmol) and pyridine (868 mg, 11 mmol) were added to the above solution. Stir at room temperature for 2 hours.
向反应液中加入饱和碳酸氢钠溶液(50毫升)。混合液用乙酸乙酯(25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：乙酸乙酯/正己烷＝1/1)，得到1.4克无色油状物2-((3-氯-5-氰基苯基)氨基)-2-氧乙酸甲酯(收率：91.2％)。MS(ESI)M/Z：239.01[M+H] +。 Saturated sodium bicarbonate solution (50 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (25 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: ethyl acetate/n-hexane=1/1) to obtain 1.4 g of colorless oily 2-((3-chloro-5-cyanophenyl)amino) Methyl-2-oxoacetate (yield: 91.2%). MS (ESI) M/Z: 239.01 [M+H] + .
步骤B：合成2-((3-氯-5-氰基苯基)氨基)-2-氧乙酸Step B: Synthesis of 2-((3-chloro-5-cyanophenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000472
Figure PCTCN2020133493-appb-000472
将2-((3-氯-5-氰基苯基)氨基)-2-氧乙酸甲酯(1.42克，5.9毫摩尔)溶于四氢呋喃/水(2/1，15毫升)的溶液中。随后，向上述溶液中加入氢氧化锂(284毫克，11.8毫摩尔)。在室温下搅拌1个小时。Methyl 2-((3-chloro-5-cyanophenyl)amino)-2-oxoacetate (1.42 g, 5.9 mmol) was dissolved in a solution of tetrahydrofuran/water (2/1, 15 mL). Subsequently, lithium hydroxide (284 mg, 11.8 mmol) was added to the above solution. Stir at room temperature for 1 hour.
向反应液中加入1M盐酸，调溶液pH值至3。混合液用乙酸乙酯(25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839毫克白色固体2-((3-氯-5-氰基苯基)氨基)-2-氧乙酸(收率：63.7％)。MS(ESI)M/Z：223.10[M-H] -。 1M hydrochloric acid was added to the reaction solution, and the pH value of the solution was adjusted to 3. The mixture was extracted with ethyl acetate (25 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. Obtained 839 mg of white solid 2-((3-chloro-5-cyanophenyl)amino)-2-oxoacetic acid (yield: 63.7%). MS(ESI) M/Z: 223.10 [MH] - .
步骤C：合成(S)-4-(2-(2-(((3-氯-5-氰基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-(((3-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzene Tert-butyl formate
Figure PCTCN2020133493-appb-000473
Figure PCTCN2020133493-appb-000473
将2-((3-氯-5-氰基苯基)氨基)-2-氧乙酸(100毫克，0.44毫摩尔)和(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(137毫克，0.4毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(230毫克，0.6毫摩尔)，N,N-二异丙基乙胺(104毫克，0.8毫摩尔)，在室温下搅拌18个小时。Combine 2-((3-chloro-5-cyanophenyl)amino)-2-oxoacetic acid (100 mg, 0.44 mmol) and (S)-4-(2-amino-3-phenylpropionamido ) Tert-Butyl benzoate (137 mg, 0.4 mmol) was dissolved in N,N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (230 mg, 0.6 mmol) was added to the above solution, N, N -Diisopropylethylamine (104 mg, 0.8 mmol), stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(25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(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/正己烷＝1/1)。得到75毫克无色油状物(S)-4-(2-(2-(((3-氯-5-氰基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：33.9％)。Saturated ammonium chloride solution (25 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 (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/n-hexane=1/1). Obtain 75 mg of colorless oil (S)-4-(2-(2-(((3-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamide Yl) tert-butyl benzoate (yield: 33.9%).
步骤D：合成(S)-4-(2-(2-(((3-氯-5-氰基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-(((3-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzene Formic acid
Figure PCTCN2020133493-appb-000474
Figure PCTCN2020133493-appb-000474
将(S)-4-(2-(2-(((3-氯-5-氰基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(75毫克，0.13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)-4-(2-(2-(((3-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) tert-butyl benzoate The ester (75 mg, 0.13 mmol) was dissolved in dichloromethane (4.0 mL). Then, 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。用甲醇(3毫升)打浆，过滤得63.5毫克白色固体(S)-4-(2-(2-(((3-氯-5-氰基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：94.2％)。MS(ESI)M/Z：547.20[M+H] +。 1H NMR(400MHz,DMSO)δ12.75(s,1H),10.52(s,1H),10.34(s,1H),9.22(d,J＝8.3Hz,1H),8.24(d,J＝1.7Hz,1H),7.91(d,J＝8.7Hz,2H),7.76(ddd,J＝18.4,17.2,8.5Hz,3H),7.37–7.24(m,3H),7.20(t,J＝7.2Hz,1H),4.78(dd,J＝15.1,7.4Hz,1H),3.19(d,J＝7.2Hz,2H). The reaction solution was concentrated under reduced pressure. Beat with methanol (3 ml) and filter to obtain 63.5 mg of white solid (S)-4-(2-(2-(((3-chloro-5-cyanophenyl)amino)-2-oxoacetamido) -3-Phenylpropionamido)benzoic acid (yield: 94.2%). MS (ESI) M/Z: 547.20 [M+H] + . 1 H NMR (400MHz, DMSO) δ 12.75 (s, 1H) ), 10.52 (s, 1H), 10.34 (s, 1H), 9.22 (d, J = 8.3 Hz, 1H), 8.24 (d, J = 1.7 Hz, 1H), 7.91 (d, J = 8.7 Hz, 2H ), 7.76 (ddd, J = 18.4, 17.2, 8.5 Hz, 3H), 7.37–7.24 (m, 3H), 7.20 (t, J = 7.2 Hz, 1H), 4.78 (dd, J = 15.1, 7.4 Hz, 1H), 3.19(d, J=7.2Hz, 2H).
实施例84Example 84
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(异烟酰胺基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Isonicotinamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000475
Figure PCTCN2020133493-appb-000475
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(异烟酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(isonicotinamido)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000476
Figure PCTCN2020133493-appb-000476
将(S)-4-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯(70毫克，115微摩尔)溶于二氯甲烷(5毫升)溶液中。随后，向上述溶液中加入异烟酰氯(19毫克，138微摩尔)和吡啶(18毫克，231微摩尔)。在室温下搅拌3小时。Add (S)-4-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxy Acetamido) propionamido) tert-butyl benzoate (70 mg, 115 micromole) was dissolved in a solution of dichloromethane (5 mL). Subsequently, isonicotinic acid chloride (19 mg, 138 micromoles) and pyridine (18 mg, 231 micromoles) were added to the above solution. Stir at room temperature for 3 hours.
将反应液过滤，滤饼用二氯甲烷(10毫升)洗涤，干燥。得到55毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(异烟酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：67.5％)。LCMS：RT＝3.91min，[M+H] +＝710.22。 The reaction solution was filtered, and the filter cake was washed with dichloromethane (10 mL) and dried. Obtain 55 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(isonicotinamido)phenyl)propionamido)benzoate (yield: 67.5%). LCMS: RT = 3.91 min, [M+H] + =710.22.
步骤B：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(异烟酰胺基)苯基)丙酰胺基)苯甲酸Step B: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(isonicotinamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000477
Figure PCTCN2020133493-appb-000477
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(异烟酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(55毫克，0.077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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Isonicotinamido)phenyl)propionamido)tert-butyl benzoate (55 mg, 0.077 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。所得残余物用甲醇(3毫升)打浆，过滤，得到23.7毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(异烟酰胺基)苯基)丙酰胺基)苯甲酸(收率：46.0％)。LCMS：RT＝3.28min，[M+H] +＝654.24。 1H NMR(500MHz,DMSO)δ10.70(s,1H),10.47(d,J＝4.9Hz,2H),9.81(s,1H),8.95(d,J＝8.2Hz,1H),8.79(d,J＝5.7Hz,2H),7.96(d,J＝2.2Hz,1H),7.89(dd,J＝9.5,7.4Hz,4H),7.78–7.72(m,1H),7.72–7.57(m,5H),7.27(d,J＝8.4Hz,2H),4.71(dd,J＝14.4,7.9Hz,1H),3.20–3.06(m,3H),1.99(dd,J＝17.4,7.6Hz,1H),1.16(t,J＝7.1Hz,1H),0.84(t,J＝6.8Hz,1H). The reaction solution was concentrated under reduced pressure. The obtained residue was slurried with methanol (3 ml) and filtered to obtain 23.7 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(isonicotinamido)phenyl)propionamido)benzoic acid (yield: 46.0%). LCMS: RT = 3.28 min, [M+H] + =654.24. 1 H NMR (500MHz, DMSO) δ 10.70 (s, 1H), 10.47 (d, J = 4.9 Hz, 2H), 9.81 (s, 1H), 8.95 (d, J = 8.2 Hz, 1H), 8.79 ( d, J = 5.7 Hz, 2H), 7.96 (d, J = 2.2 Hz, 1H), 7.89 (dd, J = 9.5, 7.4 Hz, 4H), 7.78–7.72 (m, 1H), 7.72–7.57 (m ,5H), 7.27(d,J＝8.4Hz,2H), 4.71(dd,J＝14.4,7.9Hz,1H), 3.20–3.06(m,3H),1.99(dd,J＝17.4,7.6Hz, 1H), 1.16 (t, J = 7.1Hz, 1H), 0.84 (t, J = 6.8Hz, 1H).
实施例85Example 85
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(烟酰胺基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Nicotinamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000478
Figure PCTCN2020133493-appb-000478
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(烟酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(Nicotinamido)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000479
Figure PCTCN2020133493-appb-000479
将(S)-4-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯(70毫克，115微摩尔)溶于二氯甲烷(5毫升)溶液中。随后，向上述溶液中加入异烟酰氯(19毫克，138微摩尔)和吡啶(18毫克，231微摩尔)。在室温下搅拌3小时。Add (S)-4-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxy Acetamido) propionamido) tert-butyl benzoate (70 mg, 115 micromole) was dissolved in a solution of dichloromethane (5 mL). Subsequently, isonicotinic acid chloride (19 mg, 138 micromoles) and pyridine (18 mg, 231 micromoles) were added to the above solution. Stir at room temperature for 3 hours.
将反应液过滤，滤饼用二氯甲烷(10毫升)洗涤，干燥。得到62毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(烟酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：75.4％)。LCMS：RT＝3.93min，[M+H] +＝710.22。 The reaction solution was filtered, and the filter cake was washed with dichloromethane (10 mL) and dried. Obtain 62 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(nicotinamido)phenyl)propionamido)benzoate (yield: 75.4%). LCMS: RT = 3.93 min, [M+H] + =710.22.
步骤B：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(烟酰胺基)苯基)丙酰胺基)苯甲酸Step B: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(Nicotinamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000480
Figure PCTCN2020133493-appb-000480
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(烟酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(62毫克，0.086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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Nicotinamido)phenyl)propionamido)tert-butyl benzoate (62 mg, 0.086 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。所得残余物用甲醇(3毫升)打浆，过滤，得到30.1毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(烟酰胺基)苯基)丙酰胺基)苯甲酸(收率：52.6％)。LCMS：RT＝3.30min，[M+H] +＝654.20。 1H NMR(400MHz,DMSO)δ10.70(s,1H),10.47(s,1H),10.38(s,1H),9.81(s,1H),9.08(d,J＝2.1Hz,1H),8.95(d,J＝8.3Hz,1H),8.74(dd,J＝4.8,1.6Hz,1H),8.27(d,J＝8.0Hz,1H),7.97(d,J＝2.3Hz,1H),7.90(d,J＝8.7Hz,2H),7.79–7.73(m,1H),7.72–7.52(m,6H),7.26(d,J＝8.5Hz,2H),4.71(dd,J＝14.6,7.8Hz,1H),3.13(d,J＝7.0Hz,3H),1.99(dd,J＝14.7,7.0Hz,2H),0.84(t,J＝6.8Hz,1H). The reaction solution was concentrated under reduced pressure. The obtained residue was slurried with methanol (3 ml) and filtered to obtain 30.1 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(nicotinamido)phenyl)propionamido)benzoic acid (yield: 52.6%). LCMS: RT = 3.30 min, [M+H] + =654.20. 1 H NMR (400MHz, DMSO) δ 10.70 (s, 1H), 10.47 (s, 1H), 10.38 (s, 1H), 9.81 (s, 1H), 9.08 (d, J = 2.1Hz, 1H), 8.95(d,J=8.3Hz,1H), 8.74(dd,J=4.8,1.6Hz,1H), 8.27(d,J=8.0Hz,1H), 7.97(d,J=2.3Hz,1H), 7.90(d,J=8.7Hz,2H),7.79–7.73(m,1H),7.72–7.52(m,6H),7.26(d,J=8.5Hz,2H),4.71(dd,J=14.6, 7.8Hz, 1H), 3.13 (d, J = 7.0 Hz, 3H), 1.99 (dd, J = 14.7, 7.0 Hz, 2H), 0.84 (t, J = 6.8 Hz, 1H).
实施例86Example 86
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吡啶并氨基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Pyridoamino)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000481
Figure PCTCN2020133493-appb-000481
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吡啶并氨基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(pyridoamino)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000482
Figure PCTCN2020133493-appb-000482
将(S)-4-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯(70毫克，115微摩尔)溶于二氯甲烷(5毫升)溶液中。随后，向上述溶液中加入异烟酰氯(19毫克，138微摩尔)和吡啶(18毫克，231微摩尔)。在室温下搅拌3小时。Add (S)-4-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxy Acetamido) propionamido) tert-butyl benzoate (70 mg, 115 micromole) was dissolved in a solution of dichloromethane (5 mL). Subsequently, isonicotinic acid chloride (19 mg, 138 micromoles) and pyridine (18 mg, 231 micromoles) were added to the above solution. Stir at room temperature for 3 hours.
将反应液过滤，滤饼用二氯甲烷(10毫升)洗涤，干燥。得到90毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吡啶并氨基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：109％)。LCMS：RT＝3.92min，[M+H] +＝710.22。 The reaction solution was filtered, and the filter cake was washed with dichloromethane (10 mL) and dried. Obtain 90 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(pyridoamino)phenyl)propionamido)benzoate (yield: 109%). LCMS: RT = 3.92 min, [M+H] + =710.22.
步骤B：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吡啶并氨基)苯基)丙酰胺基)苯甲酸Step B: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(pyridoamino)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000483
Figure PCTCN2020133493-appb-000483
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吡啶并氨基)苯基)丙酰胺基)苯甲酸叔丁酯(90毫克，0.126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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Pyridoamino)phenyl)propionamido)tert-butyl benzoate (90 mg, 0.126 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。所得残余物用甲醇(3毫升)打浆，过滤，得到18.8毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吡啶并氨基)苯基)丙酰胺基)苯甲酸(收率：22.0％)。LCMS：RT＝3.58min，[M+H] +＝654.44。 1H NMR(400MHz,DMSO)δ10.69(s,1H),10.56(s,1H),10.47(s,1H),9.81(d,J＝3.6Hz,1H),8.97(d,J＝8.2Hz,1H),8.72(d,J＝4.3Hz,1H),8.14(d,J＝7.8Hz,1H),8.05(td,J＝7.7,1.6Hz,1H),7.96(dd,J＝6.0,2.3Hz,1H),7.90(t,J＝6.8Hz,2H),7.83–7.72(m,3H),7.67(ddd,J＝8.5,7.5,5.0Hz,3H),7.59(dd,J＝8.6,2.4Hz,1H),7.26(d,J＝8.5Hz,2H),4.70(dd,J＝15.0,7.2Hz,1H),3.13(d,J＝7.0Hz,2H),1.99(dd,J＝14.6,6.9Hz,1H),0.84(t,J＝6.9Hz,1H). The reaction solution was concentrated under reduced pressure. The obtained residue was slurried with methanol (3 ml) and filtered to obtain 18.8 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(pyridoamino)phenyl)propionamido)benzoic acid (yield: 22.0%). LCMS: RT = 3.58 min, [M+H] + =654.44. 1 H NMR (400MHz, DMSO) δ 10.69 (s, 1H), 10.56 (s, 1H), 10.47 (s, 1H), 9.81 (d, J = 3.6 Hz, 1H), 8.97 (d, J = 8.2 Hz, 1H), 8.72 (d, J = 4.3 Hz, 1H), 8.14 (d, J = 7.8 Hz, 1H), 8.05 (td, J = 7.7, 1.6 Hz, 1H), 7.96 (dd, J = 6.0 ,2.3Hz,1H),7.90(t,J=6.8Hz,2H),7.83-7.72(m,3H),7.67(ddd,J=8.5,7.5,5.0Hz,3H),7.59(dd,J= 8.6, 2.4 Hz, 1H), 7.26 (d, J = 8.5 Hz, 2H), 4.70 (dd, J = 15.0, 7.2 Hz, 1H), 3.13 (d, J = 7.0 Hz, 2H), 1.99 (dd, J = 14.6, 6.9 Hz, 1H), 0.84 (t, J = 6.9 Hz, 1H).
实施例87Example 87
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-氧代吗啉代)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-oxomorpholino)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000484
Figure PCTCN2020133493-appb-000484
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of tert-butyl (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido)benzoate
Figure PCTCN2020133493-appb-000485
Figure PCTCN2020133493-appb-000485
将(S)-3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酸(5.0克，14.5毫摩尔)和4-氨基苯甲酸叔丁酯(2.55克，13.2毫摩尔)溶于N,N-二甲基甲酰胺(55.0毫升)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(7.5克，19.8毫摩尔)，N,N-二异丙基乙胺(3.4克，26.4毫摩尔)，在室温下搅拌18个小时。Combine (S)-3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionic acid (5.0 g, 14.5 mmol) and tert-butyl 4-aminobenzoate (2.55 g, 13.2 (Mmol) was dissolved in N,N-dimethylformamide (55.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (7.5 g, 19.8 mmol) was added to the above solution, N, N -Diisopropylethylamine (3.4 g, 26.4 mmol), stirred at room temperature for 18 hours.
向反应液中加入饱和氯化铵溶液(250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(30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＝1/10)。得到7克无色油状物(S)-4-(2-(2-(((3-氯-5-氰基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：100％)。Saturated ammonium chloride solution (250 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 (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/n-hexane=1/10). Obtain 7 grams of colorless oil (S)-4-(2-(2-(((3-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamide Yl) tert-butyl benzoate (yield: 100%).
步骤B：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(3-氧代吗啉代)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of tert-butyl (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxomorpholino)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000486
Figure PCTCN2020133493-appb-000486
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(400毫克，0.77毫摩尔)和3-吗啉酮(155毫克，1.54毫摩尔)溶于1,4-二氧六环(5.0毫升)中。随后，向上述溶液中加入碳酸铯(501毫克，1.54毫摩尔)，N 1，N 2-二甲基乙烷-1,2-二胺(136毫克，1.54毫摩尔)，碘化亚铜(146毫克，0.77毫摩尔)。在100摄氏度下搅拌18小时。 Combine (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (400 mg, 0.77 mmol) and 3-? The linone (155 mg, 1.54 mmol) was dissolved in 1,4-dioxane (5.0 mL). Subsequently, to the above solution was added cesium carbonate (501 mg, 1.54 mmol), N 1 , N 2 -dimethylethane-1,2-diamine (136 mg, 1.54 mmol), cuprous iodide ( 146 mg, 0.77 mmol). Stir at 100 degrees Celsius 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。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220毫克棕色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(3-氧代吗啉代)苯基)丙酰胺基)苯甲酸叔丁酯(收率：53.4％)。LCMS：RT＝3.90min，[M+H] +＝540.25。 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. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane = 1/2). Obtain 220 mg of brown solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxomorpholino)phenyl)propionamido)tert-butyl benzoate Ester (yield: 53.4%). LCMS: RT = 3.90 min, [M+H] + =540.25.
步骤C：合成(S)-4-(2-氨基-3-(4-(3-氧代吗啉代)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of tert-butyl (S)-4-(2-amino-3-(4-(3-oxomorpholino)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000487
Figure PCTCN2020133493-appb-000487
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(3-氧代吗啉代)苯基)丙酰胺基)苯甲酸叔丁酯(220毫克，0.4毫摩尔)溶于乙酸乙酯(3.2毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(0.8毫升,1.6毫摩尔)。在室温下搅拌18小时。Add (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(3-oxomorpholino)phenyl)propionamido) tert-butyl benzoate (220 mg , 0.4 mmol) was dissolved in ethyl acetate (3.2 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (0.8 mL, 1.6 mmol) was added to the above solution. Stir at room temperature for 18 hours.
向反应液中加入饱和碳酸氢钠溶液(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90毫克棕色固体(S)-4-(2-氨基-3-(4-(3-氧代吗啉代)苯基)丙酰胺基)苯甲酸叔丁酯(收率：53.0％)。LCMS：RT＝2.65min，[M+H] +＝440.25。 Saturated sodium bicarbonate 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. 90 mg of tert-butyl (S)-4-(2-amino-3-(4-(3-oxomorpholino)phenyl)propionamido)benzoate was obtained as a brown solid (yield: 53.0%). LCMS: RT = 2.65 min, [M+H] + =440.25.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基吗啉代)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-Hydroxymorpholino)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000488
Figure PCTCN2020133493-appb-000488
将(S)-4-(2-氨基-3-(4-(3-氧代吗啉代)苯基)丙酰胺基)苯甲酸叔丁酯(90毫克，0.2毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(60毫克，0.22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-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(205毫克，0.3毫摩尔)，N,N-二异丙基乙胺(52毫克，0.4毫摩尔)，在室温下搅拌18个小时。Combine (S)-4-(2-amino-3-(4-(3-oxomorpholino)phenyl)propionamido) tert-butyl benzoate (90 mg, 0.2 mmol) and 2-( (5-Chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (60 mg, 0.22 mmol) dissolved in N,N-dimethylformamide (3.0 mL) in. Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (205 mg, 0.3 mmol) was added to the above solution, N, N -Diisopropylethylamine (52 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92毫克无色油状物(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基吗啉代)苯基)丙酰胺基)苯甲酸叔丁酯(收率：65.2％)。LCMS：RT＝3.83min，[M+H] +＝688.25。 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 92 mg of colorless oil (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(3-Hydroxymorpholino)phenyl)propionamido)tert-butyl benzoate (yield: 65.2%). LCMS: RT = 3.83 min, [M+H] + =688.25.
步骤E：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基吗啉代)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-Hydroxymorpholino)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000489
Figure PCTCN2020133493-appb-000489
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基吗啉代)苯基)丙酰胺基)苯甲酸叔丁酯(92毫克，0.13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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 Tert-butyl -(3-hydroxymorpholino)phenyl)propionamido)benzoate (92 mg, 0.13 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11.2毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-羟基吗啉代)苯基)丙酰胺基)苯甲酸(收率：13.5％)。LCMS：RT＝3.29min，[M+H] +＝633.22。LCMS：RT＝3.29min，[M+H] +＝633.22。 1H NMR(400MHz,DMSO)δ10.69(s,1H),10.60(s,1H),9.81(s,1H),9.01(d,J＝8.3Hz,1H),7.97–7.93(m,1H),7.90(d,J＝8.7Hz,1H),7.73(dd,J＝16.4,8.7Hz,2H),7.60(dd,J＝8.6,2.4Hz,1H),7.31(s,2H),4.72(d,J＝7.6Hz,2H),3.99–3.89(m,2H),3.73–3.64(m,2H),3.16(d,J＝6.9Hz,2H). The reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 11.2 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2- Oxoacetamido)-3-(4-(3-hydroxymorpholino)phenyl)propionamido)benzoic acid (yield: 13.5%). LCMS: RT = 3.29 min, [M+H] + =633.22. LCMS: RT = 3.29 min, [M+H] + =633.22. 1 H NMR (400MHz, DMSO) δ 10.69 (s, 1H), 10.60 (s, 1H), 9.81 (s, 1H), 9.01 (d, J = 8.3 Hz, 1H), 7.97-7.93 (m, 1H) ),7.90(d,J=8.7Hz,1H),7.73(dd,J=16.4,8.7Hz,2H),7.60(dd,J=8.6,2.4Hz,1H),7.31(s,2H),4.72 (d,J=7.6Hz,2H),3.99–3.89(m,2H), 3.73–3.64(m,2H), 3.16(d,J=6.9Hz,2H).
实施例88Example 88
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吗啉-4-甲酰胺基)基)苯基)丙酰氨基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Morpholine-4-carboxamido)yl)phenyl)propionylamino)benzoic acid
Figure PCTCN2020133493-appb-000490
Figure PCTCN2020133493-appb-000490
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吗啉-4-羧酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(morpholine-4-carboxamido)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000491
Figure PCTCN2020133493-appb-000491
将叔丁基(S)-4-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸酯(70毫克，115微摩尔)溶于二氯甲烷(5毫升)溶液中。随后，向上述溶液中加入异烟酰氯(22毫克，138微摩尔)和吡啶(18毫克，231微摩尔)。在室温下搅拌3小时。Add tert-butyl(S)-4-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido)propionamido)benzoate (70 mg, 115 micromole) was dissolved in a solution of dichloromethane (5 mL). Subsequently, isonicotinic chloride (22 mg, 138 micromoles) and pyridine (18 mg, 231 micromoles) were added to the above solution. Stir at room temperature for 3 hours.
将反应液过滤，滤饼用二氯甲烷(10毫升)洗涤，干燥。得到82毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吗啉-4-羧酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：100％)。LCMS：RT＝3.73min，[M+H] +＝717.20。 The reaction solution was filtered, and the filter cake was washed with dichloromethane (10 mL) and dried. Obtain 82 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(morpholine-4-carboxamido)phenyl)propionamido)benzoate (yield: 100%). LCMS: RT = 3.73 min, [M+H] + =717.20.
步骤B：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吗啉-4-甲酰胺基)基)苯基)丙酰氨基)苯甲酸Step B: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(morpholin-4-carboxamido)yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000492
Figure PCTCN2020133493-appb-000492
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吗啉-4-羧酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(82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。(S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Morpholine-4-carboxamido)phenyl)propionamido)tert-butyl benzoate (82 mg, 0.11 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。所得残余物用甲醇(3毫升)打浆，过滤，得到31毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑 -1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(吗啉-4-甲酰胺基)基)苯基)丙酰氨基)苯甲酸(收率：40.5％)。LCMS：RT＝3.30min，[M+H] +＝662.20。 1H NMR(300MHz,DMSO)δ12.68(s,1H),10.70(s,1H),10.45(s,1H),9.81(s,1H),8.89(d,J＝8.2Hz,1H),8.44(s,1H),7.93(dd,J＝20.8,5.5Hz,3H),7.65(ddd,J＝15.8,11.0,5.5Hz,4H),7.34(d,J＝8.5Hz,2H),7.11(d,J＝8.5Hz,2H),4.65(dd,J＝14.9,7.2Hz,1H),3.64–3.53(m,4H),3.43–3.35(m,4H),3.06(d,J＝6.6Hz,2H)。 The reaction solution was concentrated under reduced pressure. The obtained residue was slurried with methanol (3 ml) and filtered to obtain 31 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(morpholin-4-carboxamido)yl)phenyl)propionylamino)benzoic acid (yield: 40.5%). LCMS: RT = 3.30 min, [M+H] + =662.20. 1 H NMR (300MHz, DMSO) δ 12.68 (s, 1H), 10.70 (s, 1H), 10.45 (s, 1H), 9.81 (s, 1H), 8.89 (d, J = 8.2 Hz, 1H), 8.44 (s, 1H), 7.93 (dd, J = 20.8, 5.5 Hz, 3H), 7.65 (ddd, J = 15.8, 11.0, 5.5 Hz, 4H), 7.34 (d, J = 8.5 Hz, 2H), 7.11 (d,J=8.5Hz,2H), 4.65(dd,J=14.9,7.2Hz,1H), 3.64–3.53(m,4H), 3.43–3.35(m,4H),3.06(d,J=6.6 Hz, 2H).
实施例89Example 89
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Piperidine-1-carboxamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000493
Figure PCTCN2020133493-appb-000493
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-羧酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000494
Figure PCTCN2020133493-appb-000494
将叔丁基(S)-4-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸酯(70毫克，115微摩尔)溶于二氯甲烷(5毫升)溶液中。随后，向上述溶液中加入1-哌啶酰氯(20毫克，138微摩尔)和吡啶(18毫克，231微摩尔)。在室温下搅拌3小时。Add tert-butyl(S)-4-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido)propionamido)benzoate (70 mg, 115 micromole) was dissolved in a solution of dichloromethane (5 mL). Subsequently, 1-pipecosyl chloride (20 mg, 138 micromoles) and pyridine (18 mg, 231 micromoles) were added to the above solution. Stir at room temperature for 3 hours.
将反应液过滤，滤饼用二氯甲烷(10毫升)洗涤，干燥。得到41毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-羧酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：100％)。LCMS：RT＝3.91min，[M+H] +＝715.20 The reaction solution was filtered, and the filter cake was washed with dichloromethane (10 mL) and dried. Obtained 41 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-(4-(piperidine-1-carboxamido)phenyl)propionamido)benzoate (yield: 100%). LCMS: RT=3.91min, [M+H] + =715.20
步骤B：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)苯甲酸Step B: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(Piperidine-1-carboxamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000495
Figure PCTCN2020133493-appb-000495
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-羧酰胺基)苯基)丙酰胺基)苯甲酸叔丁酯(41毫克，0.057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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Piperidine-1-carboxamido)phenyl)propionamido)tert-butyl benzoate (41 mg, 0.057 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。所得残余物用甲醇(3毫升)打浆，过滤，得到17.3毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(哌啶-1-甲酰胺基)苯基)丙酰胺基)苯甲酸(收率：45.3％)。LCMS：RT＝ 3.55min，[M+H] +＝660.22。 1H NMR(500MHz,DMSO)δ12.66(s,1H),10.70(s,1H),10.45(s,1H),9.83(d,J＝18.8Hz,1H),8.87(d,J＝8.2Hz,1H),8.33(s,1H),7.97(d,J＝2.3Hz,1H),7.90(d,J＝8.7Hz,2H),7.75(d,J＝8.6Hz,1H),7.68(d,J＝8.7Hz,2H),7.62–7.57(m,1H),7.33(d,J＝8.5Hz,2H),7.10(d,J＝8.5Hz,2H),4.66(dd,J＝14.9,7.2Hz,1H),3.37(dd,J＝11.5,6.1Hz,4H),3.05(d,J＝6.9Hz,2H),1.98(dt,J＝12.4,6.9Hz,1H),1.61–1.52(m,2H),1.51–1.38(m,4H),1.22(s,4H)。 The reaction solution was concentrated under reduced pressure. The obtained residue was slurried with methanol (3 mL) and filtered to obtain 17.3 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl) )Amino)-2-oxoacetamido)-3-(4-(piperidine-1-carboxamido)phenyl)propionamido)benzoic acid (yield: 45.3%). LCMS: RT = 3.55 min, [M+H] + = 660.22. 1 H NMR (500MHz, DMSO) δ 12.66 (s, 1H), 10.70 (s, 1H), 10.45 (s, 1H), 9.83 (d, J = 18.8 Hz, 1H), 8.87 (d, J = 8.2 Hz, 1H), 8.33 (s, 1H), 7.97 (d, J = 2.3 Hz, 1H), 7.90 (d, J = 8.7 Hz, 2H), 7.75 (d, J = 8.6 Hz, 1H), 7.68 ( d,J=8.7Hz,2H), 7.62–7.57(m,1H), 7.33(d,J=8.5Hz,2H), 7.10(d,J=8.5Hz,2H), 4.66(dd,J=14.9 ,7.2Hz,1H), 3.37(dd,J=11.5,6.1Hz,4H),3.05(d,J=6.9Hz,2H),1.98(dt,J=12.4,6.9Hz,1H),1.61-1.52 (m, 2H), 1.51–1.38 (m, 4H), 1.22 (s, 4H).
实施例90Example 90
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧酰胺基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000496
Figure PCTCN2020133493-appb-000496
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Cyanopiperidine-1-carboxamido)phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000497
Figure PCTCN2020133493-appb-000497
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)苯甲酸叔丁酯(68毫克，0.093毫摩尔)和盐酸哌啶-4-甲腈(27毫克，0.18毫摩尔)溶于乙腈(5.0毫升)的溶液中。随后，向上述溶液中加入N,N-二异丙基乙胺(39毫克，0.3毫摩尔)在室温下搅拌5小时。(S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((Phenoxycarbonyl)amino)phenyl)propionamido)tert-butyl benzoate (68 mg, 0.093 mmol) and piperidine-4-carbonitrile hydrochloride (27 mg, 0.18 mmol) dissolved in acetonitrile ( 5.0 ml) of the solution. Subsequently, N,N-diisopropylethylamine (39 mg, 0.3 mmol) was added to the above solution and stirred at room temperature for 5 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22毫克无色油状物(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基丙酰胺基)苯甲酸叔丁酯(收率：35.4％)。LCMS：RT＝3.85min，[M+H] +＝740.21 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 22 mg of colorless oil (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(4-Cyanopiperidine-1-carboxamido)phenylpropionamido)tert-butyl benzoate (yield: 35.4%). LCMS: RT = 3.85 min, [M+ H] + ＝740.21
步骤B：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧酰胺基)苯基)丙酰胺基)苯甲酸Step B: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-Cyanopiperidine-1-carboxamido)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000498
Figure PCTCN2020133493-appb-000498
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-甲酰胺基)苯基丙酰胺基)苯甲酸叔丁酯(22毫克，0.029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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-Cyanopiperidine-1-carboxamido)phenylpropionamido) tert-butyl benzoate (22 mg, 0.029 mmol) was dissolved in dichloromethane (4.0 mL). Then, the above solution Trifluoroacetic acid (1.0 mL) was added to the mixture and stirred at room temperature for 2 hours.
将反应液减压浓缩。所得残余物用乙酸乙酯(3毫升)打浆，过滤，得到3.9毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-氰基哌啶-1-羧酰胺基)苯基)丙酰胺基)苯甲酸(收率：20.0％)。LCMS：RT＝3.40min，[M+H] +＝685.21 The reaction solution was concentrated under reduced pressure. The resulting residue was slurried with ethyl acetate (3 mL) and filtered to obtain 3.9 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl )Phenyl)amino)-2-oxoacetamido)-3-(4-(4-cyanopiperidine-1-carboxamido)phenyl)propionamido)benzoic acid (yield: 20.0% ). LCMS: RT = 3.40 min, [M+H] + =685.21
实施例91Example 91
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000499
Figure PCTCN2020133493-appb-000499
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-异丙基哌嗪-2-酮Step A: Synthesis of 4-isopropylpiperazin-2-one
Figure PCTCN2020133493-appb-000500
Figure PCTCN2020133493-appb-000500
将哌嗪-2-酮(500毫克，5毫摩尔)和丙酮(580毫克，10毫摩尔)溶于甲醇(10毫升)中.随后向上述溶液中加入醋酸(0.5毫升)和氰基硼氢化钠(627毫克，10毫摩尔)。在50摄氏度下搅拌18小时。Piperazine-2-one (500 mg, 5 mmol) and acetone (580 mg, 10 mmol) were dissolved in methanol (10 mL). Then acetic acid (0.5 mL) and cyanoborohydride were added to the above solution Sodium (627 mg, 10 mmol). Stir at 50 degrees Celsius for 18 hours.
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250毫克棕色固体4-异丙基哌嗪-2-酮(收率：23.1％)。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). 250 mg of brown solid 4-isopropylpiperazin-2-one was obtained (yield: 23.1%). LCMS: RT=0.83 min, [M+H] + =143.11.
步骤B：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propane Amido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000501
Figure PCTCN2020133493-appb-000501
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(300毫克，0.57毫摩尔)和4-异丙基哌嗪-2-酮(164毫克，1.14毫摩尔)溶于甲苯(15.0毫升)中。随后，向上述溶液中加入碳酸铯(376毫克，1.14毫摩尔)，N 1，N 2-二甲基乙烷-1,2-二胺(101毫克，1.14毫摩尔)，碘化亚铜109毫克，0.57毫摩尔)。在110摄氏度下搅拌18小时。 Combine (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (300 mg, 0.57 mmol) and 4-iso Propylpiperazin-2-one (164 mg, 1.14 mmol) was dissolved in toluene (15.0 mL). Subsequently, cesium carbonate (376 mg, 1.14 mmol), N 1 , N 2 -dimethylethane-1,2-diamine (101 mg, 1.14 mmol), and cuprous iodide 109 were added to the above solution. Mg, 0.57 mmol). Stir at 110 degrees Celsius 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(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：乙酸乙酯/正己烷＝1/1)。得到180毫克棕色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：31.8％)。LCMS：RT＝3.53min，[M+H] +＝580.33。 Saturated ammonium chloride solution (10 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: ethyl acetate/n-hexane=1/1). Obtain 180 mg of brown solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl) Propanamido) tert-butyl benzoate (yield: 31.8%). LCMS: RT=3.53 min, [M+H] + =580.33.
步骤C：合成(S)-4-(2-氨基-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of tert-butyl (S)-4-(2-amino-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000502
Figure PCTCN2020133493-appb-000502
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(180毫克，0.31毫摩尔)溶于乙酸乙酯(2.5毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(0.6毫升,1.2毫摩尔)。在室温下搅拌4小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido) Tert-butyl benzoate (180 mg, 0.31 mmol) was dissolved in ethyl acetate (2.5 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (0.6 mL, 1.2 mmol) was added to the above solution. Stir at room temperature for 4 hours.
向反应液中加入饱和碳酸氢钠溶液(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70毫克棕色固体(S)-4-(2-氨基-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：47.0％)。Saturated sodium bicarbonate 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 70 mg of brown solid (S)-4-(2-amino-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate (Yield: 47.0%).
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-isopropyl-2-oxopiperazin-1-yl)phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000503
Figure PCTCN2020133493-appb-000503
将(S)-4-(2-氨基-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(70毫克，0.14毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(50毫克，0.18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-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸盐(85毫克，0.22毫摩尔)，N,N-二异丙基乙胺(40毫克，0.3毫摩尔)，在室温下搅拌18个小时。(S)-4-(2-amino-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (70 mg, 0.14 mmol) and 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (50 mg, 0.18 mmol) dissolved in N,N-di Methylformamide (3.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (85 mg, 0.22 mmol) was added to the above solution, N, N -Diisopropylethylamine (40 mg, 0.3 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70毫克无色油状物(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基丙酰胺基)苯甲酸叔丁酯(收率：66.3％)。LCMS：RT＝3.50min，[M+H] +＝729.33。 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 70 mg of colorless oil (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenylpropionamido)tert-butyl benzoate (yield: 66.3%). LCMS: RT = 3.50 min , [M+H] + =729.33.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000504
Figure PCTCN2020133493-appb-000504
将(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(70毫克，0.09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)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( Tert-butyl 4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoate (70 mg, 0.09 mmol) was dissolved in dichloromethane (4.0 mL). Subsequently, trifluoroacetic acid (1.0 mL) was added to the above solution and stirred 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5.1毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸(收率：8.2％)。LCMS：RT＝2.84min，[M+H] +＝674.27。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC to obtain 5.1 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid (yield: 8.2%). LCMS: RT = 2.84 min, [M+H] + =674.27.
实施例92Example 92
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(环丙基甲基)-2-氧代哌嗪-1-基)苯基丙酰胺基)苯甲酸三氟乙酸盐Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(Cyclopropylmethyl)-2-oxopiperazin-1-yl)phenylpropionamido)benzoic acid trifluoroacetate
Figure PCTCN2020133493-appb-000505
Figure PCTCN2020133493-appb-000505
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(环丙基甲基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(cyclopropylmethyl)-2-oxopiperazin-1-yl)benzene (Yl) propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000506
Figure PCTCN2020133493-appb-000506
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(300毫克，0.57毫摩尔)和4-(环丙基甲基)哌嗪-2-酮(178毫克，1.14毫摩尔)溶于甲苯(15.0毫升)中。随后，向上述溶液中加入碳酸铯(376毫克，1.14毫摩尔)，N 1，N 2-二甲基乙烷-1,2-二胺(101毫克，1.14毫摩尔)，碘化亚铜109毫克，0.57毫摩尔)。在110摄氏度下搅拌18小时。 Combine (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (300 mg, 0.57 mmol) and 4-( Cyclopropylmethyl)piperazin-2-one (178 mg, 1.14 mmol) was dissolved in toluene (15.0 mL). Subsequently, cesium carbonate (376 mg, 1.14 mmol), N 1 , N 2 -dimethylethane-1,2-diamine (101 mg, 1.14 mmol), and cuprous iodide 109 were added to the above solution. Mg, 0.57 mmol). Stir at 110 degrees Celsius 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(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毫克棕色固体(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(环丙基甲基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：45.2％)。LCMS：RT＝3.63min，[M+H] +＝592.21。 Saturated ammonium chloride solution (10 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 brown solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(cyclopropylmethyl)-2-oxopiperazin-1-yl) (Phenyl) propionamido) tert-butyl benzoate (yield: 45.2%). LCMS: RT = 3.63 min, [M+H] + =592.21.
步骤B：合成(S)-4-(2-氨基-3-(4-(4-(环丙基甲基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-amino-3-(4-(4-(cyclopropylmethyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzene Tert-butyl formate
Figure PCTCN2020133493-appb-000507
Figure PCTCN2020133493-appb-000507
将(S)-4-(2-((叔丁氧羰基)氨基)-3-(4-(4-(环丙基甲基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(260毫克，0.44毫摩尔)溶于乙酸乙酯(3.5毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(0.87毫升,1.7毫摩尔)。在室温下搅拌4小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(cyclopropylmethyl)-2-oxopiperazin-1-yl)phenyl)propane Tert-butyl amido)benzoate (260 mg, 0.44 mmol) was dissolved in ethyl acetate (3.5 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (0.87 mL, 1.7 mmol) was added to the above solution. Stir at room temperature for 4 hours.
向反应液中加入饱和碳酸氢钠溶液(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40毫克棕色固体(S)-4-(2-氨基-3-(4-(4-(环丙基甲基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：18.3％)。Saturated sodium bicarbonate 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 40 mg of brown solid (S)-4-(2-amino-3-(4-(4-(cyclopropylmethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) Tert-Butyl benzoate (yield: 18.3%).
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(环丙基甲基)-2-氧哌嗪-1-基)苯基丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Cyclopropylmethyl)-2-oxopiperazin-1-yl)phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000508
Figure PCTCN2020133493-appb-000508
将(S)-4-(2-氨基-3-(4-(4-(环丙基甲基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(40毫克，0.08毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(24毫克，0.09毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(46毫克，0.12毫摩尔)，N,N-二异丙基乙胺(21毫克，0.16毫摩尔)，在室温下搅拌18个小时。Add (S)-4-(2-amino-3-(4-(4-(cyclopropylmethyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate Ester (40 mg, 0.08 mmol) and 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (24 mg, 0.09 mmol) are dissolved in N,N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (46 mg, 0.12 mmol) was added to the above solution, N, N -Diisopropylethylamine (21 mg, 0.16 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40毫克无色油状物(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-异丙基-2-氧代哌嗪-1-基)苯基丙酰胺基)苯甲酸叔丁酯(收率：65.0％)。LCMS：RT＝3.52min，[M+H] +＝741.28。 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 40 mg of colorless oil (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(4-isopropyl-2-oxopiperazin-1-yl)phenylpropionamido)tert-butyl benzoate (yield: 65.0%). LCMS: RT=3.52min , [M+H] + =741.28.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(环丙基甲基)-2-氧哌嗪-1-基)苯基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Cyclopropylmethyl)-2-oxopiperazin-1-yl)phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000509
Figure PCTCN2020133493-appb-000509
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(环丙基甲基)-2-氧哌嗪-1-基)苯基丙酰胺基)苯甲酸叔丁酯(40毫克，0.05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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(Cyclopropylmethyl)-2-oxopiperazin-1-yl)phenylpropionamido) tert-butyl benzoate (40 mg, 0.05 mmol) dissolved in dichloromethane (4.0 mL) Then, trifluoroacetic acid (1.0 mL) was added to the above solution and stirred 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毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(环丙基甲基)-2-氧哌嗪-1-基)苯基丙酰胺基)苯甲酸三氟乙酸盐(收率：92.4％)。LCMS：RT＝2.92min，[M+H] +＝685.27。 1H NMR(400MHz,DMSO)δ12.78(s,1H),10.72(s,1H),10.48(s,1H),9.82(s,1H),9.04(d,J＝8.1Hz,1H),7.95(d,J＝2.3Hz,1H),7.91(d,J＝8.7Hz,2H),7.76(d,J＝8.6Hz,1H),7.69(d,J＝8.8Hz,2H),7.61(dd,J＝8.6,2.4Hz,1H),7.34(d,J＝8.5Hz,2H),7.27(d,J＝8.5Hz,2H),5.75(s,1H),4.72(dd,J＝14.2,7.9Hz,1H),3.18(d,J＝6.7Hz,3H),1.12(d,J＝16.1Hz,1H),0.66(d,J＝6.8Hz,2H),0.39(s,2H)。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC to obtain 34 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2- Oxoacetamido)-3-(4-(4-(cyclopropylmethyl)-2-oxopiperazin-1-yl)phenylpropionamido)benzoic acid trifluoroacetate (Yield: 92.4%). LCMS: RT = 2.92 min, [M+H] + =685.27. 1 H NMR (400MHz, DMSO) δ 12.78 (s, 1H), 10.72 (s, 1H), 10.48 (s, 1H) , 9.82 (s, 1H), 9.04 (d, J = 8.1 Hz, 1H), 7.95 (d, J = 2.3 Hz, 1H), 7.91 (d, J = 8.7 Hz, 2H), 7.76 (d, J = 8.6Hz, 1H), 7.69 (d, J = 8.8 Hz, 2H), 7.61 (dd, J = 8.6, 2.4 Hz, 1H), 7.34 (d, J = 8.5 Hz, 2H), 7.27 (d, J = 8.5Hz, 2H), 5.75 (s, 1H), 4.72 (dd, J = 14.2, 7.9 Hz, 1H), 3.18 (d, J = 6.7 Hz, 3H), 1.12 (d, J = 16.1 Hz, 1H) , 0.66 (d, J = 6.8 Hz, 2H), 0.39 (s, 2H).
实施例93Example 93
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸三氟乙酸盐Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid trifluoroacetate
Figure PCTCN2020133493-appb-000510
Figure PCTCN2020133493-appb-000510
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(4-甲基环己基)哌嗪-2-酮Step A: Synthesis of 4-(4-methylcyclohexyl)piperazin-2-one
Figure PCTCN2020133493-appb-000511
Figure PCTCN2020133493-appb-000511
将哌嗪-2-酮(500毫克，5毫摩尔)和4-甲氧基环己-1-酮(1.28克，10毫摩尔)溶于甲醇(10毫升)中.随后向上述溶液中加入醋酸(0.5毫升)和氰基硼氢化钠(627毫克，10毫摩尔)。在50摄氏度下搅拌18小时。Piperazine-2-one (500 mg, 5 mmol) and 4-methoxycyclohexan-1-one (1.28 g, 10 mmol) were dissolved in methanol (10 mL). Then added to the above solution Acetic acid (0.5 mL) and sodium cyanoborohydride (627 mg, 10 mmol). Stir at 50 degrees Celsius for 18 hours.
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100毫克棕色固体4-(4-甲基环己基)哌嗪-2-酮(收率：10.7％)。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). 100 mg of brown solid 4-(4-methylcyclohexyl)piperazin-2-one was obtained (yield: 10.7%). LCMS: RT=0.95 min, [M+H] + =213.15.
步骤B：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazine-1 -Yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000512
Figure PCTCN2020133493-appb-000512
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(500毫克，1毫摩尔)和4-(4-甲基环己基)哌嗪-2-酮(170毫克，0.8毫摩尔)溶于甲苯(15.0毫升)中。随后，向上述溶液中加入碳酸铯(522毫克，1.6毫摩尔)，N 1，N 2-二甲基乙烷-1,2-二胺(141毫克，1.6毫摩尔)，碘化亚铜109毫克，0.8毫摩尔)。在110摄氏度下搅拌18小时。 Combine (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (500 mg, 1 mmol) and 4-( 4-Methylcyclohexyl)piperazin-2-one (170 mg, 0.8 mmol) was dissolved in toluene (15.0 mL). Subsequently, cesium carbonate (522 mg, 1.6 mmol), N 1 , N 2 -dimethylethane-1,2-diamine (141 mg, 1.6 mmol), and cuprous iodide 109 were added to the above solution. Mg, 0.8 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(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280毫克棕色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：53.6％)。LCMS：RT＝3.65min，[M+H] +＝650.13。 Saturated ammonium chloride solution (30 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 280 mg of brown solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazine- 1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 53.6%). LCMS: RT = 3.65 min, [M+H] + =650.13.
步骤C：合成(S)-4-(2-氨基-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido )Tert-Butyl Benzoate
Figure PCTCN2020133493-appb-000513
Figure PCTCN2020133493-appb-000513
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(280毫克，0.43毫摩尔)溶于乙酸乙酯(16毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(4毫升)。在室温下搅拌4小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl) (Phenyl) propionamido) tert-butyl benzoate (280 mg, 0.43 mmol) was dissolved in ethyl acetate (16 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (4 mL) was added to the above solution. Stir at room temperature for 4 hours.
将反应液减压浓缩。得到200毫克棕色固体(S)-4-(2-氨基-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：84.2％)。The reaction solution was concentrated under reduced pressure. Obtain 200 mg of brown solid (S)-4-(2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamide Yl) tert-butyl benzoate (yield: 84.2%).
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000514
Figure PCTCN2020133493-appb-000514
将(S)-4-(2-氨基-3-(4-(4-(4-甲氧基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(200毫克，0.36毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(116毫克，0.43毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(207毫克，0.54毫摩尔)，N,N-二异丙基乙胺(94毫克，0.72毫摩尔)，在室温下搅拌18个小时。(S)-4-(2-amino-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid Tert-butyl ester (200 mg, 0.36 mmol) and 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (116 mg, 0.43 mmol) Dissolve in N,N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (207 mg, 0.54 mmol) was added to the above solution, N, N -Diisopropylethylamine (94 mg, 0.72 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。得到150毫克无色油状物(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：55.1％)。LCMS：RT＝3.60min，[M+H] +＝799.21。 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 150 mg of colorless oil (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 55.1%). LCMS: RT = 3.60 min, [M+H] + =799.21.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000515
Figure PCTCN2020133493-appb-000515
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(150毫克，0.18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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(4-Methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (150 mg, 0.18 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。所得残余物用制备HPLC纯化得到33毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基丙酰胺基)苯甲酸三氟乙酸盐(收率：23.5％)。LCMS：RT＝2.89min，[M+H] +＝744.29。 1H NMR(400MHz,DMSO)δ10.72(s,1H),10.48(s,1H),9.84(d,J＝12.6Hz,1H),9.05(d,J＝8.1Hz,1H),7.93(dd,J＝17.5,5.5Hz,3H),7.76(d,J＝8.6Hz,1H),7.68(d,J＝8.8Hz,2H),7.62(dd,J＝8.6,2.3Hz,1H),7.34(d,J＝8.5Hz,2H),7.27(d,J＝8.5Hz,2H),4.72(dd,J＝14.2,8.0Hz,1H),3.23(d,J＝8.0Hz,3H),2.10(s,2H),1.85(s,1H),1.65(s,1H),1.44(dd,J＝31.0,16.6Hz,3H),1.27–1.10(m,2H)。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC to obtain 33 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(4-(4-methoxycyclohexyl)-2-oxopiperazin-1-yl)phenylpropionamido)benzoic acid trifluoroacetate ( Yield: 23.5%). LCMS: RT = 2.89 min, [M+H] + =744.29. 1 H NMR (400MHz, DMSO) δ 10.72 (s, 1H), 10.48 (s, 1H), 9.84 (d ,J=12.6Hz,1H),9.05(d,J=8.1Hz,1H),7.93(dd,J=17.5,5.5Hz,3H),7.76(d,J=8.6Hz,1H),7.68(d ,J=8.8Hz,2H), 7.62(dd,J=8.6,2.3Hz,1H), 7.34(d,J=8.5Hz,2H), 7.27(d,J=8.5Hz,2H), 4.72(dd ,J=14.2,8.0Hz,1H), 3.23(d,J=8.0Hz,3H), 2.10(s,2H), 1.85(s,1H), 1.65(s,1H), 1.44(dd,J= 31.0, 16.6 Hz, 3H), 1.27–1.10 (m, 2H).
实施例94Example 94
合成4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基丙酰胺基)苯甲酸Synthesis of 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(Hydroxymethyl)-2-oxopiperidin-1-yl)phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000516
Figure PCTCN2020133493-appb-000516
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-((2S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of 4-((2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenyl ) Propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000517
Figure PCTCN2020133493-appb-000517
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(500毫克，1毫摩尔)和4-(4-甲基环己基)哌嗪-2-酮(103毫克，0.8毫摩尔)溶于甲苯(15.0毫升)中。随后，向上述溶液中加入碳酸铯(522毫克，1.6毫摩尔)，N 1，N 2-二甲基乙烷-1,2-二胺(141毫克，1.6毫摩尔)，碘化亚铜109毫克，0.8毫摩尔)。在110摄氏度下搅拌18小时。 Combine (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (500 mg, 1 mmol) and 4-( 4-Methylcyclohexyl)piperazin-2-one (103 mg, 0.8 mmol) was dissolved in toluene (15.0 mL). Subsequently, cesium carbonate (522 mg, 1.6 mmol), N 1 , N 2 -dimethylethane-1,2-diamine (141 mg, 1.6 mmol), and cuprous iodide 109 were added to the above solution. Mg, 0.8 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(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75毫克棕色固体4-((2S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：13.6％)。LCMS：RT＝3.70min，[M+H] +＝567.30。 Saturated ammonium chloride solution (30 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 75 mg of brown solid 4-((2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)benzene (Yl)propionamido)tert-butyl benzoate (yield: 13.6%). LCMS: RT = 3.70 min, [M+H] + =567.30.
步骤B：合成4-((2S)-2-氨基-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of tertiary 4-((2S)-2-amino-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido)benzoic acid Butyl
Figure PCTCN2020133493-appb-000518
Figure PCTCN2020133493-appb-000518
将4-((2S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(70毫克，0.13毫摩尔)溶于乙酸乙酯(16毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(4毫升)。在室温下搅拌4小时。4-((2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamide Tert-Butyl benzoate (70 mg, 0.13 mmol) was dissolved in ethyl acetate (16 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (4 mL) was added to the above solution. Stir at room temperature for 4 hours.
将反应液减压浓缩。得到60毫克棕色固体4-((2S)-2-氨基-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：98.1％)。The reaction solution was concentrated under reduced pressure. Obtain 60 mg of brown solid 4-((2S)-2-amino-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido)benzoic acid Tert-Butyl ester (yield: 98.1%).
步骤C：合成4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Hydroxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000519
Figure PCTCN2020133493-appb-000519
将4-((2S)-2-氨基-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(60毫克，0.12毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(41毫克，0.15毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(73毫克，0.19毫摩尔)，N,N-二异丙基乙胺(31毫克，0.24毫摩尔)，在室温下搅拌18个小时。Add 4-((2S)-2-amino-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido) tert-butyl benzoate ( 60 mg, 0.12 mmol) and 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (41 mg, 0.15 mmol) dissolved in N, N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (73 mg, 0.19 mmol) was added to the above solution, N, N -Diisopropylethylamine (31 mg, 0.24 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。得到22毫克无色油状物4-((2S)-2-(2-((5-氯 -2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：24.0％)。LCMS：RT＝3.65min，[M+H] +＝716.30。 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 22 mg of colorless oil 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido ) Tert-butyl-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido)benzoate (yield: 24.0%). LCMS: RT = 3.65 min, [M+H] + =716.30.
步骤D：合成4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基丙酰胺基)苯甲酸Step D: Synthesis of 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000520
Figure PCTCN2020133493-appb-000520
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲氧基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(22毫克，0.03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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(4-Methoxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (22 mg, 0.03 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。所得残余物用制备HPLC纯化得到9.7毫克白色固体4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基丙酰胺基)苯甲酸(收率：4.4％)。LCMS：RT＝2.96min，[M+H] +＝660.30。 1H NMR(500MHz,DMSO)δ11.74(s,1H),10.15(s,1H),9.78(s,1H),8.01(s,1H),7.95(d,J＝1.8Hz,1H),7.80(dt,J＝8.7,5.4Hz,2H),7.37(dt,J＝19.8,8.6Hz,5H),7.08(t,J＝4.1Hz,1H),5.40(s,1H),3.49–3.31(m,4H),3.25(dd,J＝9.9,6.2Hz,4H),3.15(dd,J＝14.4,10.0Hz,2H),2.14(d,J＝12.3Hz,2H),2.02(d,J＝12.8Hz,1H),1.89(s,1H),1.70(s,1H),1.55(s,1H),1.45(t,J＝13.0Hz,2H)。 The reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 9.7 mg of white solid 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2- Oxoacetamido)-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenylpropionamido)benzoic acid (yield: 4.4%). LCMS: RT = 2.96 min, [M+H] + = 660.30. 1 H NMR (500MHz, DMSO) δ 11.74 (s, 1H), 10.15 (s, 1H), 9.78 (s, 1H), 8.01 (s, 1H) ), 7.95 (d, J = 1.8 Hz, 1H), 7.80 (dt, J = 8.7, 5.4 Hz, 2H), 7.37 (dt, J = 19.8, 8.6 Hz, 5H), 7.08 (t, J = 4.1 Hz ,1H),5.40(s,1H),3.49–3.31(m,4H), 3.25(dd,J＝9.9,6.2Hz,4H), 3.15(dd,J＝14.4,10.0Hz,2H), 2.14( d, J = 12.3 Hz, 2H), 2.02 (d, J = 12.8 Hz, 1H), 1.89 (s, 1H), 1.70 (s, 1H), 1.55 (s, 1H), 1.45 (t, J = 13.0 Hz, 2H).
实施例95Example 95
合成4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基丙酰胺基)苯甲酸Synthesis of 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(Hydroxymethyl)-2-oxopiperidin-1-yl)phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000521
Figure PCTCN2020133493-appb-000521
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-(羟甲基)哌啶-2-酮Step A: Synthesis of 4-(hydroxymethyl)piperidin-2-one
Figure PCTCN2020133493-appb-000522
Figure PCTCN2020133493-appb-000522
将2-氧过哌啶-4-羧酸甲酯(500毫克，3.18毫摩尔)溶于甲醇(15.0毫升)中。随后，向上述溶液中加入硼氢化锂(138毫克，6.36毫摩尔)。在50摄氏度下搅拌1小时。Methyl 2-oxoperpiperidine-4-carboxylate (500 mg, 3.18 mmol) was dissolved in methanol (15.0 mL). Subsequently, lithium borohydride (138 mg, 6.36 mmol) was added to the above solution. Stir at 50 degrees Celsius for 1 hour.
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450毫克棕色固体4-(羟甲基)哌啶-2-酮(收率：109％)。LCMS：RT＝0.98min，[M+H] +＝130.10。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 450 mg of brown solid 4-(hydroxymethyl)piperidin-2-one was obtained (yield: 109%). LCMS: RT=0.98 min, [M+H] + =130.10.
步骤B：合成(2-氧代哌啶-4-基)4-甲基苯磺酸甲酯Step B: Synthesis of (2-oxopiperidin-4-yl) 4-methylbenzenesulfonate methyl ester
Figure PCTCN2020133493-appb-000523
Figure PCTCN2020133493-appb-000523
将4-(羟甲基)哌啶-2-酮(450毫克，3.4毫摩尔)溶于乙腈(15.0毫升)中。随后，向上述溶液中加入三乙胺(705毫克，6.9毫摩尔)和4-甲基苯磺酰氯(1.33克，6.9毫摩尔)。在室温下搅拌18小时。4-(Hydroxymethyl)piperidin-2-one (450 mg, 3.4 mmol) was dissolved in acetonitrile (15.0 mL). Subsequently, triethylamine (705 mg, 6.9 mmol) and 4-methylbenzenesulfonyl chloride (1.33 g, 6.9 mmol) were added to the above solution. Stir at room temperature for 18 hours.
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90毫克棕色油状物4-(羟甲基)哌啶-2-酮(收率：9.8％)。LCMS：RT＝1.33min，[M+H] +＝284.10。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 90 mg of brown oily 4-(hydroxymethyl)piperidin-2-one was obtained (yield: 9.8%). LCMS: RT=1.33 min, [M+H] + =284.10.
步骤C：合成4-(乙基甲基)哌啶-2-酮Step C: Synthesis of 4-(ethylmethyl)piperidin-2-one
Figure PCTCN2020133493-appb-000524
Figure PCTCN2020133493-appb-000524
将(2-氧代哌啶-4-基)4-甲基苯磺酸甲酯(90毫克，0.3毫摩尔)溶于乙醇(5.0毫升)中。随后，向上述溶液中加入氢化钠(15毫克，0.6毫摩尔)。在70摄氏度下搅拌18小时。(2-oxopiperidin-4-yl)methyl 4-methylbenzenesulfonate (90 mg, 0.3 mmol) was dissolved in ethanol (5.0 mL). Subsequently, sodium hydride (15 mg, 0.6 mmol) was added to the above solution. Stir at 70 degrees Celsius for 18 hours.
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180毫克棕色固体4-(羟甲基)哌啶-2-酮(收率：190％)。LCMS：RT＝1.16min，[M+H] +＝157.10。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol/dichloromethane = 1/10). 180 mg of brown solid 4-(hydroxymethyl)piperidin-2-one was obtained (yield: 190%). LCMS: RT=1.16 min, [M+H] + =157.10.
步骤D：合成4-((2S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(乙氧基甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of 4-((2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(ethoxymethyl)-2-oxopiperidin-1-yl) (Phenyl) propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000525
Figure PCTCN2020133493-appb-000525
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(320毫克，0.6毫摩尔)和4-(乙基甲基)哌啶-2-酮(193毫克，1.2毫摩尔)溶于甲苯(15.0毫升)中。随后，向上述溶液中加入碳酸铯(401毫克，1.2毫摩尔)，N 1，N 2-二甲基乙烷-1,2-二胺(108毫克，1.2毫摩尔)，碘化亚铜117毫克，0.6毫摩尔)。在110摄氏度下搅拌18小时。 (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (320 mg, 0.6 mmol) and 4-( Ethylmethyl)piperidin-2-one (193 mg, 1.2 mmol) was dissolved in toluene (15.0 mL). Subsequently, cesium carbonate (401 mg, 1.2 mmol), N 1 , N 2 -dimethylethane-1,2-diamine (108 mg, 1.2 mmol), and cuprous iodide 117 were added to the above solution. Mg, 0.6 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(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140毫克棕色固体4-((2S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(乙氧基甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：38.0％)。LCMS：RT＝3.98min，[M+H] +＝596.43。 Saturated ammonium chloride solution (30 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 140 mg of brown solid 4-((2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(ethoxymethyl)-2-oxopiperidin-1-yl )Phenyl)propionamido)tert-butyl benzoate (yield: 38.0%). LCMS: RT = 3.98 min, [M+H] + =596.43.
步骤E：合成4-((2S)-2-氨基-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of tertiary 4-((2S)-2-amino-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido)benzoic acid Butyl
Figure PCTCN2020133493-appb-000526
Figure PCTCN2020133493-appb-000526
将4-((2S)-2-((叔丁氧羰基)氨基)-3-(4-(4-(乙氧基甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(140毫克，0.21毫摩尔)溶于乙酸乙酯(16毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(4毫升)。在室温下搅拌4小时。Add 4-((2S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4-(ethoxymethyl)-2-oxopiperidin-1-yl)phenyl) Tert-butyl propionamido)benzoate (140 mg, 0.21 mmol) was dissolved in ethyl acetate (16 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (4 mL) was added to the above solution. Stir at room temperature for 4 hours.
将反应液减压浓缩。得50毫克棕色固体4-((2S)-2-氨基-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：42.5％)。The reaction solution was concentrated under reduced pressure. Obtain 50 mg of brown solid 4-((2S)-2-amino-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido)benzoic acid Tert-Butyl ester (yield: 42.5%).
步骤F：合成4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(乙氧基甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step F: Synthesis of 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(ethoxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000527
Figure PCTCN2020133493-appb-000527
将4-((2S)-2-氨基-3-(4-(4-(羟甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(50毫克，0.1毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(29毫克，0.11毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(57毫克，0.15毫摩尔)，N,N-二异丙基乙胺(26毫克，0.2毫摩尔)，在室温下搅拌18个小时。Add 4-((2S)-2-amino-3-(4-(4-(hydroxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido) tert-butyl benzoate ( 50 mg, 0.1 mmol) and 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (29 mg, 0.11 mmol) dissolved in N, N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (57 mg, 0.15 mmol) was added to the above solution, N, N -Diisopropylethylamine (26 mg, 0.2 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。得到6毫克无色油状物4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(乙氧基甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：8.7％)。LCMS：RT＝3.93min，[M+H] +＝744.33。 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. Obtain 6 mg of colorless oil 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(4-(ethoxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 8.7%). LCMS: RT = 3.93 min, [M+H] + =744.33.
步骤G：合成4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(乙氧基甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸Step G: Synthesis of 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(ethoxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000528
Figure PCTCN2020133493-appb-000528
将4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(乙氧基甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(6毫克，0.008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 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(Ethoxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido) tert-butyl benzoate (6 mg, 0.008 mmol) dissolved in dichloromethane (4.0 Milliliters). 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2.8毫克白色固体4-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(乙氧基甲基)-2-氧代哌啶-1-基)苯基)丙酰胺基)苯甲酸(收率：42.1％)。LCMS：RT＝3.64min，[M+H] +＝689.20 The reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 2.8 mg of white solid 4-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2- Oxoacetamido)-3-(4-(4-(ethoxymethyl)-2-oxopiperidin-1-yl)phenyl)propionamido)benzoic acid (yield: 42.1%) . LCMS: RT = 3.64 min, [M+H] + =689.20
实施例96Example 96
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000529
Figure PCTCN2020133493-appb-000529
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 PCTCN2020133493-appb-000530
Figure PCTCN2020133493-appb-000530
将哌嗪-2-酮(500毫克，5毫摩尔)和4-羟基环己-1-酮(1.1克，10毫摩尔)溶于甲醇(10毫升)中.随后向上述溶液中加入醋酸(0.5毫升)和氰基硼氢化钠(627毫克，10毫摩尔)。在室温下搅拌18小时。Piperazine-2-one (500 mg, 5 mmol) and 4-hydroxycyclohexan-1-one (1.1 g, 10 mmol) were dissolved in methanol (10 mL). Then acetic acid ( 0.5 mL) and sodium cyanoborohydride (627 mg, 10 mmol). Stir at room temperature for 18 hours.
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毫克棕色固体4-(4-羟基环己基)哌嗪-2-酮(收率：90％)。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). 400 mg of brown solid 4-(4-hydroxycyclohexyl)piperazin-2-one was obtained (yield: 90%). LCMS: RT=0.82, [M+H] + =199.14.
步骤B：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl )Phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000531
Figure PCTCN2020133493-appb-000531
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(200毫克，0.38毫摩尔)和4-(4-羟基环己基)哌嗪-2-酮(152毫克，0.77毫摩尔)溶于甲苯(15.0毫升)中。随后，向上述溶液中加入碳酸铯(250毫克，0.77毫摩尔)，N 1，N 2-二甲基乙烷-1,2-二胺(67毫克，0.77毫摩尔)，碘化亚铜73毫克，0.38毫摩尔)。在110摄氏度下搅拌18小时。 Combine (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (200 mg, 0.38 mmol) and 4-( 4-Hydroxycyclohexyl)piperazin-2-one (152 mg, 0.77 mmol) was dissolved in toluene (15.0 mL). Subsequently, cesium carbonate (250 mg, 0.77 mmol), N 1 , N 2 -dimethylethane-1,2-diamine (67 mg, 0.77 mmol), and cuprous iodide were added to the above solution. Mg, 0.38 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(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160毫克棕色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：38.0％)。LCMS：RT＝3.13min，[M+H] +＝636.35 Saturated ammonium chloride solution (30 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). To obtain 160 mg of brown solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazine-1- (Yl)phenyl)propionamido)tert-butyl benzoate (yield: 38.0%). LCMS: RT = 3.13 min, [M+H] + =636.35
步骤C：合成(S)-4-(2-氨基-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-amino-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzene Tert-butyl formate
Figure PCTCN2020133493-appb-000532
Figure PCTCN2020133493-appb-000532
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(160毫克，0.25毫摩尔)溶于乙酸乙酯(16毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(4毫升)。在室温下搅拌4小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl ) Propanamido) tert-butyl benzoate (160 mg, 0.25 mmol) was dissolved in ethyl acetate (16 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (4 mL) was added to the above solution. Stir at room temperature for 4 hours.
将反应液减压浓缩。得40毫克棕色固体(S)-4-(2-氨基-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：33.4％)。The reaction solution was concentrated under reduced pressure. Obtain 40 mg of brown solid (S)-4-(2-amino-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido) Tert-butyl benzoate (yield: 33.4%).
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-羟基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000533
Figure PCTCN2020133493-appb-000533
将(S)-4-(2-氨基-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(40毫克，0.07毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(21毫克，0.08毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(42毫克，0.11毫摩尔)，N,N-二异丙基乙胺(19毫克，0.15毫摩尔)，在室温下搅拌18个小时。Add (S)-4-(2-amino-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate Ester (40 mg, 0.07 mmol) and 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (21 mg, 0.08 mmol) are dissolved in N,N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (42 mg, 0.11 mmol) was added to the above solution, N, N -Diisopropylethylamine (19 mg, 0.15 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。得到46毫克无色油状物(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-羟基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：83.2％)。LCMS：RT＝3.06min，[M+H] +＝785.31。 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. Obtained 46 mg of colorless oil (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)tert-butyl benzoate (yield: 83.2%). LCMS: RT = 3.06 min, [M+H] + =785.31.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸三氟乙酸盐Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid trifluoroacetate
Figure PCTCN2020133493-appb-000534
Figure PCTCN2020133493-appb-000534
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-羟基环己基)-2-氧哌嗪-1-基)苯基)丙酰胺基)苯甲酸叔丁酯(46毫克，0.058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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(4-Hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido) tert-butyl benzoate (46 mg, 0.058 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。所得残余物用制备HPLC纯化得到14毫克白色固体(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-羟基环己基)-2-氧代哌嗪-1-基)苯基)丙酰胺基)苯甲酸三氟乙酸盐(收率：33.6％)。LCMS：RT＝2.79min，[M+H] +＝728.35。 1H NMR(400MHz,DMSO)δ12.75(s,1H),10.72(s,1H),10.48(s,1H),9.82(s,1H),9.05(d,J＝8.0Hz,1H),7.97(d,J＝2.3Hz,1H),7.92(d,J＝8.7Hz,2H),7.78(d,J＝8.6Hz,1H),7.70(d,J＝8.8Hz,2H),7.63(dd,J＝8.6,2.4Hz,1H),7.38–7.32(m,2H),7.28(dd,J＝8.5,2.8Hz,2H)，4.72(dd,J＝13.8,7.6Hz,1H),3.17(s,2H),2.53(s,1H),2.09–1.87(m,2H),1.79(s,2H),1.46(s,2H),1.18(dd,J＝19.3,12.2Hz,2H),0.83(dd,J＝8.9,6.6Hz,1H)。 The reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 14 mg of white solid (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(4-(4-hydroxycyclohexyl)-2-oxopiperazin-1-yl)phenyl)propionamido)benzoic acid trifluoroacetate ( Yield: 33.6%). LCMS: RT = 2.79 min, [M+H] + = 728.35. 1 H NMR (400MHz, DMSO) δ 12.75 (s, 1H), 10.72 (s, 1H), 10.48 (s) ,1H),9.82(s,1H),9.05(d,J=8.0Hz,1H),7.97(d,J=2.3Hz,1H),7.92(d,J=8.7Hz,2H),7.78(d ,J=8.6Hz,1H), 7.70(d,J=8.8Hz,2H), 7.63(dd,J=8.6,2.4Hz,1H), 7.38–7.32(m,2H), 7.28(dd,J= 8.5,2.8Hz,2H), 4.72(dd,J=13.8,7.6Hz,1H),3.17(s,2H),2.53(s,1H),2.09–1.87(m,2H),1.79(s,2H) ), 1.46 (s, 2H), 1.18 (dd, J = 19.3, 12.2 Hz, 2H), 0.83 (dd, J = 8.9, 6.6 Hz, 1H).
实施例97Example 97
合成(S)-4-(3-([[1,1'-联苯]-4-基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)苯甲酸Synthesis of (S)-4-(3-([[1,1'-biphenyl]-4-yl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl) (Phenyl)amino)-2-oxoacetamido)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000535
Figure PCTCN2020133493-appb-000535
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(3-([[1,1'-联苯]-4-基]-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of tert-butyl (S)-4-(3-([[1,1'-biphenyl]-4-yl]-2-((tert-butoxycarbonyl)amino)propionamido)benzoate
Figure PCTCN2020133493-appb-000536
Figure PCTCN2020133493-appb-000536
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(500毫克，0.96毫摩尔)和苯硼酸(140毫克，1.4毫摩尔)溶于N,N-二甲基甲酰胺(15.0毫升)中。随后，向上述溶液中加入碳酸钾399毫克，1.93毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(78毫克，0.09毫摩尔)。在100摄氏度下搅拌18小时。Combine (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (500 mg, 0.96 mmol) and phenylboronic acid ( 140 mg, 1.4 mmol) was dissolved in N,N-dimethylformamide (15.0 mL). Subsequently, 399 mg of potassium carbonate, 1.93 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (78 mg, 0.09 mmol) were added to the above solution. Stir at 100 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(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270毫克棕色固体(S)-4-(3-([[1,1'-联苯]-4-基]-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(收率：38.0％)。Saturated ammonium chloride solution (30 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 270 mg of brown solid (S)-4-(3-([[1,1'-biphenyl]-4-yl]-2-((tert-butoxycarbonyl)amino)propionamido)tert-butyl benzoate Ester (yield: 38.0%).
步骤B：合成(S)-4-(3-([(1,1'-联苯]-4-基)-2-氨基丙酰胺)苯甲酸叔丁酯Step B: Synthesis of (S)-4-(3-([(1,1'-biphenyl]-4-yl)-2-aminopropionamide) tert-butyl benzoate
Figure PCTCN2020133493-appb-000537
Figure PCTCN2020133493-appb-000537
将(S)-4-(3-([[1,1'-联苯]-4-基]-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(270毫克，0.52毫摩尔)溶于乙酸乙酯(8毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(2毫升)。在室温下搅拌4小时。Add (S)-4-(3-([[1,1'-biphenyl]-4-yl]-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (270 mg , 0.52 mmol) was dissolved in ethyl acetate (8 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (2 mL) was added to the above solution. The mixture was stirred at room temperature for 4 hours.
将反应液减压浓缩。得220毫克棕色固体(S)-4-(3-([(1,1'-联苯]-4-基)-2-氨基丙酰胺)苯甲酸叔丁酯(收率：100％)。The reaction solution was concentrated under reduced pressure. 220 mg of tert-butyl (S)-4-(3-([(1,1'-biphenyl]-4-yl)-2-aminopropionamide)benzoate was obtained as a brown solid (yield: 100%).
步骤C：合成(S)-4-(3-([[1,1'-联苯]-4-基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(3-([[1,1'-biphenyl]-4-yl)-2-(2-((5-chloro-2-(1H-tetrazole-1 -Yl)phenyl)amino)-2-oxoacetamido)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000538
Figure PCTCN2020133493-appb-000538
将(S)-4-(3-([(1,1'-联苯]-4-基)-2-氨基丙酰胺)苯甲酸叔丁酯(220毫克，0.5毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(170毫克，0.6毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(300毫克，0.76毫摩尔)，N,N-二异丙基乙胺(136毫克，1.02毫摩尔)，在室温下搅拌18个小时。Combine (S)-4-(3-([(1,1'-biphenyl]-4-yl)-2-aminopropionamide) tert-butyl benzoate (220 mg, 0.5 mmol) and 2-( (5-Chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (170 mg, 0.6 mmol) dissolved in N,N-dimethylformamide (5.0 mL) Then, add 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (300 mg, 0.76 mmol) to the above solution, N , N-Diisopropylethylamine (136 mg, 1.02 mmol), stirred at room temperature 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(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(15毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到180毫克无色油状物(S)-4-(3-([[1,1'-联苯]-4-基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)苯甲酸叔丁酯(收率：54.3％)。LCMS：RT＝4.53min，[M-H] -＝663.27。 Saturated ammonium chloride solution (30 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 (15 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. To obtain 180 mg of colorless oil (S)-4-(3-([[1,1'-biphenyl]-4-yl)-2-(2-((5-chloro-2-(1H-tetra (Azol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)tert-butyl benzoate (yield: 54.3%). LCMS: RT = 4.53 min, [MH] - = 663.27.
步骤D：合成(S)-4-(3-([[1,1'-联苯]-4-基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(3-([[1,1'-biphenyl]-4-yl)-2-(2-((5-chloro-2-(1H-tetrazole-1 -Yl)phenyl)amino)-2-oxoacetamido)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000539
Figure PCTCN2020133493-appb-000539
将(S)-4-(3-([[1,1'-联苯]-4-基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)苯甲酸叔丁酯(180毫克，0.27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)-4-(3-([[1,1'-biphenyl]-4-yl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl) Phenyl)amino)-2-oxoacetamido)propionamido)tert-butyl benzoate (180 mg, 0.27 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14毫克白色固体(S)-4-(3-([[1,1'-联苯]-4-基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)苯甲酸(收率：8.5％)。LCMS：RT＝4.11min，[M-H] -＝608.08。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC to obtain 14 mg of white solid (S)-4-(3-([[1,1'-biphenyl]-4-yl)-2-(2-((5-chloro-2 -(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)benzoic acid (yield: 8.5%). LCMS: RT = 4.11 min, [MH] - =608.08.
实施例98Example 98
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(Furan-3-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000540
Figure PCTCN2020133493-appb-000540
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of tert-butyl (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(furan-3-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000541
Figure PCTCN2020133493-appb-000541
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(500毫克，0.96毫摩尔)和呋喃-3-基硼酸(129毫克，1.1毫摩尔)溶于N,N-二甲基甲酰胺(15.0毫升)中。随后，向上述溶液中加入碳酸钾266毫克，1.93毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(70毫克，0.09毫摩尔)。在100摄氏度下搅拌18小时。Combine (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (500 mg, 0.96 mmol) and furan-3 Boronic acid (129 mg, 1.1 mmol) was dissolved in N,N-dimethylformamide (15.0 mL). Subsequently, potassium carbonate 266 mg, 1.93 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (70 mg, 0.09 mmol) were added to the above solution. Stir at 100 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(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370毫克棕色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：76.3％)。Saturated ammonium chloride solution (30 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 370 mg of brown solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(furan-3-yl)phenyl)propionamido) tert-butyl benzoate ( Yield: 76.3%).
步骤B：合成(S)-4-(2-氨基-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of tert-butyl (S)-4-(2-amino-3-(4-(furan-3-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000542
Figure PCTCN2020133493-appb-000542
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸叔丁酯(200毫克，0.40毫摩尔)溶于乙酸乙酯(8毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(2毫升)。在室温下搅拌4小时。(S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(furan-3-yl)phenyl)propionamido) tert-butyl benzoate (200 mg, 0.40 (Mmol) was dissolved in ethyl acetate (8 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (2 mL) was added to the above solution. Stir at room temperature for 4 hours.
将反应液减压浓缩。得130毫克棕色固体(S)-4-(2-氨基-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：80.2％)。The reaction solution was concentrated under reduced pressure. 130 mg of tert-butyl (S)-4-(2-amino-3-(4-(furan-3-yl)phenyl)propionamido)benzoate was obtained as a brown solid (yield: 80.2%).
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(furan-3-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000543
Figure PCTCN2020133493-appb-000543
将(S)-4-(2-氨基-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸叔丁酯(130毫克，0.32毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(102毫克，0.38毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(182毫克，0.48毫摩尔)，N,N-二异丙基乙胺(82毫克，0.6毫摩尔)，在室温下搅拌18个小时。Combine (S)-4-(2-amino-3-(4-(furan-3-yl)phenyl)propionamido) tert-butyl benzoate (130 mg, 0.32 mmol) and 2-((5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (102 mg, 0.38 mmol) was dissolved in N,N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (182 mg, 0.48 mmol) was added to the above solution, N,N -Diisopropylethylamine (82 mg, 0.6 mmol), stirred at room temperature 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(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(15毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到150毫克无色油状物(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：71.8％)。LCMS：RT＝4.14min，[M-H] -＝653.20。 Saturated ammonium chloride solution (30 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 (15 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 150 mg of colorless oil (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(furan-3-yl)phenyl)propionamido)tert-butyl benzoate (yield: 71.8%). LCMS: RT = 4.14 min, [MH] - =653.20.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(furan-3-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000544
Figure PCTCN2020133493-appb-000544
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸叔丁酯(150毫克，0.22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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 Tert-butyl -(furan-3-yl)phenyl)propionamido)benzoate (150 mg, 0.22 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5.2毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-3-基)苯基)丙酰胺基)苯甲酸(收率：3.9％)。LCMS：RT＝3.93min，[M-H] -＝598.08。 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC to obtain 5.2 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2- Oxoacetamido)-3-(4-(furan-3-yl)phenyl)propionamido)benzoic acid (yield: 3.9%). LCMS: RT = 3.93 min, [MH] - =598.08.
实施例99Example 99
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(furan-2-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000545
Figure PCTCN2020133493-appb-000545
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of tert-butyl (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(furan-2-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000546
Figure PCTCN2020133493-appb-000546
将(S)-4-(3-(4-溴苯基)-2-((叔丁氧羰基)氨基)丙酰胺基)苯甲酸叔丁酯(500毫克，0.96毫摩尔)和呋喃-2-基硼酸(129毫克，1.1毫摩尔)溶于N,N-二甲基甲酰胺(15.0毫升)中。随后，向上述溶液中加入碳酸钾266毫克，1.93毫摩尔)，[1,1'-双(二苯基膦)二茂铁]二氯化钯(70毫克，0.09毫摩尔)。在100摄氏度下搅拌18小时。Combine (S)-4-(3-(4-bromophenyl)-2-((tert-butoxycarbonyl)amino)propionamido) tert-butyl benzoate (500 mg, 0.96 mmol) and furan-2 Boronic acid (129 mg, 1.1 mmol) was dissolved in N,N-dimethylformamide (15.0 mL). Subsequently, potassium carbonate 266 mg, 1.93 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (70 mg, 0.09 mmol) were added to the above solution. Stir at 100 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(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420毫克棕色固体(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：86.1％)。Saturated ammonium chloride solution (30 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 420 mg of brown solid (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(furan-2-yl)phenyl)propionamido) tert-butyl benzoate ( Yield: 86.1%).
步骤B：合成(S)-4-(2-氨基-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of tert-butyl (S)-4-(2-amino-3-(4-(furan-2-yl)phenyl)propionamido)benzoate
Figure PCTCN2020133493-appb-000547
Figure PCTCN2020133493-appb-000547
将(S)-4-(2-((叔丁氧基羰基)氨基)-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(250毫克，0.49毫摩尔)溶于乙酸乙酯(8毫升)中。随后，向上述溶液中加入2摩尔/升的盐酸乙酸乙酯溶液(2毫升)。在室温下搅拌4小时。Add (S)-4-(2-((tert-butoxycarbonyl)amino)-3-(4-(furan-2-yl)phenyl)propionamido) tert-butyl benzoate (250 mg, 0.49 (Mmol) was dissolved in ethyl acetate (8 mL). Subsequently, a 2 mol/L hydrochloric acid ethyl acetate solution (2 mL) was added to the above solution. Stir at room temperature for 4 hours.
将反应液减压浓缩。得170毫克棕色固体(S)-4-(2-氨基-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：85.2％)。The reaction solution was concentrated under reduced pressure. 170 mg of tert-butyl (S)-4-(2-amino-3-(4-(furan-2-yl)phenyl)propionamido)benzoate was obtained as a brown solid (yield: 85.2%).
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(furan-2-yl)phenyl)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000548
Figure PCTCN2020133493-appb-000548
将(S)-4-(2-氨基-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(170毫克，0.4毫摩尔)和2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(123毫克，0.4毫摩尔)溶于N,N-二甲基甲酰胺(5.0毫升)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(238毫克，0.6毫摩尔)，N,N-二异丙基乙胺(108毫克，0.8毫摩尔)，在室温下搅拌18个小时。(S)-4-(2-amino-3-(4-(furan-2-yl)phenyl)propionamido) tert-butyl benzoate (170 mg, 0.4 mmol) and 2-((5 -Chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (123 mg, 0.4 mmol) was dissolved in N,N-dimethylformamide (5.0 mL). Subsequently, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (238 mg, 0.6 mmol) was added to the above solution, N, N -Diisopropylethylamine (108 mg, 0.8 mmol), stirred at room temperature 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(20毫升×3次)萃取。合并有机相。有机相先用饱和食盐水(15毫升×3次)，然后用无水硫酸钠干燥，最后减压浓缩。得到180毫克无色油状物(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(收率：68.0％)。LCMS：RT＝4.13min，[M-H] -＝653.20。 Saturated ammonium chloride solution (30 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 (15 ml×3 times), then dried over anhydrous sodium sulfate, and finally concentrated under reduced pressure. Obtain 180 mg of colorless oil (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(furan-2-yl)phenyl)propionamido)tert-butyl benzoate (yield: 68.0%). LCMS: RT = 4.13 min, [MH] - =653.20.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(furan-2-yl)phenyl)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000549
Figure PCTCN2020133493-appb-000549
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸叔丁酯(180毫克，0.27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)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 Tert-butyl -(furan-2-yl)phenyl)propionamido)benzoate (180 mg, 0.27 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8.7毫克白色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(呋喃-2-基)苯基)丙酰胺基)苯甲酸(收率：5.3％)。LCMS：RT＝3.92min，[M-H] -＝598.08 The reaction solution was concentrated under reduced pressure. The obtained residue was purified by preparative HPLC to obtain 8.7 mg of white solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2- Oxoacetamido)-3-(4-(furan-2-yl)phenyl)propionamido)benzoic acid (yield: 5.3%). LCMS: RT = 3.92 min, [MH] - =598.08
实施例100Example 100
合成(S)-4-(2-(2-((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000550
Figure PCTCN2020133493-appb-000550
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成4-氯-1-环丁氧基-2-硝基苯Step A: Synthesis of 4-chloro-1-cyclobutoxy-2-nitrobenzene
Figure PCTCN2020133493-appb-000551
Figure PCTCN2020133493-appb-000551
室温下，将4-氯-1-氟-2-硝基苯(800.0毫克，4.56毫摩尔)、环丁醇(394.0毫克，5.47毫摩尔)和碳酸铯(3.0克，9.12毫摩尔)加入N,N-二甲基甲酰胺(20.0毫升)中，N 2保护下，50摄氏度反应1.5小时。 At room temperature, 4-chloro-1-fluoro-2-nitrobenzene (800.0 mg, 4.56 mmol), cyclobutanol (394.0 mg, 5.47 mmol) and cesium carbonate (3.0 g, 9.12 mmol) were added to N , N-dimethylformamide (20.0 ml), under N 2 protection, react at 50 degrees Celsius for 1.5 hours.
反应结束，垫无水硫酸钠抽滤，滤饼用50.0毫升乙酸乙酯洗涤，滤液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(50毫升×2次)洗涤，然后用无水硫酸钠干燥，最后硅胶柱层析纯化(洗脱剂：正己烷：乙酸乙酯＝20：1)得到460.2毫克淡黄色固体4-氯-1-环丁氧基-2-硝基苯(收率：44.0％)。LCMS:RT＝4.31min,[M+H] +＝228.14。 After the reaction was over, a pad of anhydrous sodium sulfate was suction filtered, the filter cake was washed with 50.0 ml ethyl acetate, the filtrate was extracted with ethyl acetate (30 ml × 3 times), the organic phases were combined, and the organic phase was first washed with saturated brine (50 ml × 2 times) washing, then drying with anhydrous sodium sulfate, and finally silica gel column chromatography (eluent: n-hexane: ethyl acetate = 20:1) to obtain 460.2 mg of pale yellow solid 4-chloro-1-cyclobutoxide 2-nitrobenzene (yield: 44.0%). LCMS: RT = 4.31 min, [M+H] + = 228.14.
步骤B：合成5-氯-2-环丁氧基苯胺Step B: Synthesis of 5-chloro-2-cyclobutoxyaniline
Figure PCTCN2020133493-appb-000552
Figure PCTCN2020133493-appb-000552
室温下，将4-氯-1-环丁氧基-2-硝基苯(460.0毫克，2.02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(4.6克，20.21毫摩尔)，N 2保护下，室温反应2小时。 At room temperature, 4-chloro-1-cyclobutoxy-2-nitrobenzene (460.0 mg, 2.02 mmol) was added to ethyl acetate, and stannous chloride dihydrate (4.6 G, 20.21 mmol), under N 2 protection, react at room temperature for 2 hours.
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50毫升乙酸乙酯洗涤，滤液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(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：正己烷：乙酸乙酯＝20：1)。得到349.0毫克乳白色固体5-氯-2-环丁氧基苯胺(收率：87.0％)。LCMS:RT＝4.17min,[M+H] +＝198.14。 After the reaction was completed, it was quenched with saturated sodium bicarbonate aqueous solution, and excess sodium bicarbonate was added to adjust the pH to weakly alkaline. The filter cake was filtered with Celite pad and the filter cake was washed with 50 ml of ethyl acetate. The filtrate was washed with ethyl acetate ( 30 ml×3 times), the organic phases were combined, and the organic phase was washed with saturated brine (50 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: n-hexane: ethyl acetate = 20:1). Obtained 349.0 mg of creamy white solid 5-chloro-2-cyclobutoxyaniline (yield: 87.0%). LCMS: RT = 4.17 min, [M+H] + =198.14.
步骤C：合成2-((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000553
Figure PCTCN2020133493-appb-000553
N 2保护、冰浴下，向含有5-氯-2-环丁氧基苯胺(349.0毫克，1.77毫摩尔)和吡啶(280.0毫克，3.54毫摩尔)的二氯甲烷(5.0毫升)中滴加2-氯-2-氧代乙酸甲酯(258.7毫克，2.21毫摩尔)，滴毕，室温反应30分钟。 Under N 2 protection and ice bath, add dropwise to dichloromethane (5.0 ml) containing 5-chloro-2-cyclobutoxyaniline (349.0 mg, 1.77 mmol) and pyridine (280.0 mg, 3.54 mmol) Methyl 2-chloro-2-oxoacetate (258.7 mg, 2.21 mmol) was dripped and reacted at room temperature for 30 minutes.
反应结束，加水淬灭，混合液用二氯甲烷(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×3次)洗涤，然后用无水硫酸钠干燥，最后硅胶柱层析纯化(洗脱剂：正己烷：乙酸乙酯＝20：1)得到356.0毫克白色固体2-((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酸甲酯(收率：87.0％)。LCMS:RT＝3.86min,[M-H] -＝282.12。 After the reaction was over, it was quenched by adding water, and the mixture was extracted with dichloromethane (50 ml×3 times). The organic phases were combined, and the organic phase was washed with saturated brine (30 ml×3 times), then dried over anhydrous sodium sulfate, and finally purified by silica gel column chromatography (eluent: n-hexane: ethyl acetate = 20:1) Obtained 356.0 mg of white solid methyl 2-((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetate (yield: 87.0%). LCMS: RT = 3.86 min, [MH] - =282.12.
步骤D：合成2-((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酸Step D: Synthesis of 2-((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000554
Figure PCTCN2020133493-appb-000554
冰浴下，向含有((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酸甲酯(356.0毫克，1.25毫摩尔)的四氢呋喃(14.0毫升)中滴加氢氧化锂(105.0毫克，2.50毫摩尔)水溶液(7.0毫升)，滴毕即反应结束。Under ice bath, to ((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxyacetic acid methyl ester (356.0 mg, 1.25 mmol) in tetrahydrofuran (14.0 ml) was added dropwise lithium hydroxide (105.0 mg, 2.50 mmol) aqueous solution (7.0 ml), the reaction is complete when the dropping is 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(30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，最后减压浓缩得到333.0毫克淡黄色固体合成2-((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酸，无需纯化，直接用于下一步反应(收率：98.8％)。LCMS:RT＝4.08min,[M-H] -＝268.33。 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 (30 ml×3 times), combine the organic phases, and use saturated brine (30 Ml×2 times), then dried with anhydrous sodium sulfate, and finally concentrated under reduced pressure to obtain 333.0 mg of pale yellow solid synthetic 2-((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetic acid , Without purification, directly used in the next reaction (yield: 98.8%). LCMS: RT = 4.08 min, [MH] - =268.33.
步骤E：合成4-(2-(2-(((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of 4-(2-(2-(((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) tert-benzoic acid Butyl
Figure PCTCN2020133493-appb-000555
Figure PCTCN2020133493-appb-000555
室温下，将2-((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酸(100.0毫克，0.37毫摩尔)、(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(115.0毫克，0.34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(192.0毫克，0.51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(87.0毫克，0.67毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 2-((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetic acid (100.0 mg, 0.37 mmol), (S)-4-(2-amino-3- Phenylpropionamido) tert-butyl benzoate (115.0 mg, 0.34 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate Salt (192.0 mg, 0.51 mmol) was added to N,N-dimethylformamide (5.0 mL), and N,N-diisopropylethylamine (87.0 mg, 0.67 mmol) was added dropwise. 2 Under protection, react at room temperature for 5 hours.
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＝5/1)。得到129.0毫克黄色固体4-(2-(2-(((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：40.9％)。LCMS:RT＝4.33min,[M-H] -＝590.43。 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=5/1). Obtain 129.0 mg of yellow solid 4-(2-(2-(((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid Tert-butyl ester (yield: 40.9%). LCMS: RT = 4.33 min, [MH] - =590.43.
步骤F：合成(S)-4-(2-(2-(((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-(((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetamido)-3-phenylpropionamido )benzoic acid
Figure PCTCN2020133493-appb-000556
Figure PCTCN2020133493-appb-000556
室温下，将4-(2-(2-(((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(65.0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2小时。At room temperature, the 4-(2-(2-(((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) benzoic acid tert Butyl ester (65.0 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 2 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23.6毫克白色固体合成(S)-4-(2-(2-(((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：40.0％)。LCMS:RT＝3.25min,[M+H] +＝536.69。 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 23.6 mg of white solid synthetic (S)-4-(2-(2-(((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetamide (Yl)-3-phenylpropionamido)benzoic acid (yield: 40.0%). LCMS: RT = 3.25 min, [M+H] + =536.69.
实施例101Example 101
合成(S)-4-(3-(4-([[1,4'-联哌啶]-1'-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-酰基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸Synthesis of (S)-4-(3-(4-([[1,4'-bipiperidine]-1'-carboxamido)phenyl)-2-(2-((5-chloro-2- (1H-tetrazole-1-acyl)phenyl)amino)-2-oxoacetamido)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000557
Figure PCTCN2020133493-appb-000557
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(3-(4-(1,4'-双哌啶)-1'-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of (S)-4-(3-(4-(1,4'-bispiperidine)-1'-carboxamido)phenyl)-2-(2-((5-chloro-2 -(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)tert-butyl benzoate
Figure PCTCN2020133493-appb-000558
Figure PCTCN2020133493-appb-000558
室温下，将(S)-4-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)苯甲酸叔丁酯(50.0毫克，0.08毫摩尔)、[1,4'-联哌啶]-1'-羰基氯盐酸盐(26.5毫克，0.10毫摩尔)和吡啶(20.0毫克，0.25毫摩尔)加入二氯甲烷(3.0毫升)中，N 2保护下，室温反应5小时。 At room temperature, (S)-4-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido) propionamido) tert-butyl benzoate (50.0 mg, 0.08 mmol), [1,4'-bipiperidine]-1'-carbonyl chloride hydrochloride (26.5 mg, 0.10 (20.0 mg, 0.25 mmol) and pyridine (20.0 mg, 0.25 mmol) were added to dichloromethane (3.0 mL) and reacted at room temperature for 5 hours under N 2 protection.
反应结束，加水淬灭，混合液用二氯甲烷(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：二氯甲烷/甲醇＝10/1)。得到49.0毫克黄色固体(S)-4-(3-(4-(((1,4'-双哌啶)-1'-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)苯甲酸叔丁酯(收率：71.9％)。LCMS:RT＝4.28min,[M-H] -＝797.63。 After the reaction was completed, it was quenched by adding water, the mixed solution was extracted with dichloromethane (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: dichloromethane/methanol=10/1). Obtain 49.0 mg of yellow solid (S)-4-(3-(4-(((1,4'-bispiperidine)-1'-carboxamido)phenyl)-2-(2-((5- Chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)tert-butyl benzoate (yield: 71.9%). LCMS: RT = 4.28 min, [MH] - = 797.63.
步骤B：合成(S)-4-(3-(4-(((1,4'-双哌啶]-1'-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)苯甲酸Step B: Synthesis of (S)-4-(3-(4-(((1,4'-bispiperidine)-1'-carboxamido)phenyl)-2-(2-((5-chloro -2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)benzoic acid
Figure PCTCN2020133493-appb-000559
Figure PCTCN2020133493-appb-000559
室温下，将(S)-4-(3-(4-(((1,4'-双哌啶)-1'-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰 胺基)苯甲酸叔丁酯(49.0毫克，0.06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2小时。At room temperature, (S)-4-(3-(4-(((1,4'-bispiperidine)-1'-carboxamido)phenyl)-2-(2-((5-chloro -2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)tert-butyl benzoate (49.0 mg, 0.06 mmol) was added to dichloromethane (5.0 mL) In, trifluoroacetic acid (1.0 ml) was added dropwise, and the reaction was carried out at room temperature for 2 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42.3毫克白色固体合成(S)-4-(3-(4-(((1,4'-双哌啶)-1'-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)苯甲酸叔丁酯(收率：92.0％)。LCMS:RT＝3.04min,[M-H] -＝743.27。 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 42.3 mg of white solid synthetic (S)-4-(3-(4-(((1,4'-bispiperidine)-1'-carboxamido)phenyl)- Tert-Butyl 2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)benzoate (Yield: 92.0% ). LCMS: RT = 3.04 min, [MH] - =743.27.
实施例102Example 102
合成(S)-4-(2-(2-((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropane Amido) benzoic acid
Figure PCTCN2020133493-appb-000560
Figure PCTCN2020133493-appb-000560
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成1-(4-氯-2-硝基苯基)-1H-咪唑Step A: Synthesis of 1-(4-chloro-2-nitrophenyl)-1H-imidazole
Figure PCTCN2020133493-appb-000561
Figure PCTCN2020133493-appb-000561
室温下，将5-氯-2-氟硝基苯(2.0克，11.40毫摩尔)、1H-咪唑(892.0毫克，131.10毫摩尔)和碳酸钾(1.9克，13.70毫摩尔)加入N,N-二甲基甲酰胺(14.0毫升)中，N 2保护下，90摄氏度反应1.5小时。 At room temperature, 5-chloro-2-fluoronitrobenzene (2.0 g, 11.40 mmol), 1H-imidazole (892.0 mg, 131.10 mmol) and potassium carbonate (1.9 g, 13.70 mmol) were added to N,N- In dimethylformamide (14.0 ml), under the protection of N 2 , the reaction was carried out at 90°C for 1.5 hours.
反应结束，垫无水硫酸钠抽滤，滤饼用50.0毫升乙酸乙酯洗涤，滤液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(50毫升×2次)洗涤，然后用无水硫酸钠干燥，最后用硅胶柱层析纯化(洗脱剂：正己烷/乙酸乙酯＝1/1)得到791.0毫克棕色油状物1-(4-氯-2-硝基苯基)-1H-咪唑。LCMS:RT＝4.37min,[M+H] +＝224.15。 After the reaction was over, a pad of anhydrous sodium sulfate was suction filtered, the filter cake was washed with 50.0 ml ethyl acetate, the filtrate was extracted with ethyl acetate (30 ml × 3 times), the organic phases were combined, and the organic phase was first washed with saturated brine (50 ml × 2 times), then dried with anhydrous sodium sulfate, and finally purified by silica gel column chromatography (eluent: n-hexane/ethyl acetate = 1/1) to obtain 791.0 mg of brown oily 1-(4-chloro-2 -Nitrophenyl)-1H-imidazole. LCMS: RT = 4.37 min, [M+H] + =224.15.
步骤B：合成5-氯-2-(1H-咪唑-1-基)苯胺Step B: Synthesis of 5-chloro-2-(1H-imidazol-1-yl)aniline
Figure PCTCN2020133493-appb-000562
Figure PCTCN2020133493-appb-000562
室温下，将1-(4-氯-2-硝基苯基)-1H-咪唑(741.0毫克，3.31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(7.5克，33.12毫摩尔)，N 2保护下，室温反应2小时。 At room temperature, add 1-(4-chloro-2-nitrophenyl)-1H-imidazole (741.0 mg, 3.31 mmol) into ethyl acetate. Under ice bath, add stannous chloride dihydrate in batches (7.5 g, 33.12 mmol), under N 2 protection, react at room temperature for 2 hours.
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50毫升乙酸乙酯洗涤，滤液用乙酸乙酯(30毫升×3次)萃取，合并有机相，有机相先用饱和食盐水(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：正己烷：乙酸乙酯＝1：1)。得到569.0毫克乳白色固体5-氯-2-环丁氧基苯胺(收率：88.6％)。LCMS:RT＝4.22min,[M+H] +＝194.17。 After the reaction was completed, it was quenched with saturated sodium bicarbonate aqueous solution, and excess sodium bicarbonate was added to adjust the pH to weakly alkaline. The filter cake was filtered with Celite pad and the filter cake was washed with 50 ml of ethyl acetate. The filtrate was washed with ethyl acetate ( 30 ml×3 times), the organic phases were combined, and the organic phase was washed with saturated brine (50 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: n-hexane: ethyl acetate=1:1). Obtained 569.0 mg of creamy white solid 5-chloro-2-cyclobutoxyaniline (yield: 88.6%). LCMS: RT = 4.22 min, [M+H] + = 194.17.
步骤C：合成2-((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酸甲酯Step C: Synthesis of methyl 2-((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000563
Figure PCTCN2020133493-appb-000563
N 2保护、冰浴下，向含有5-氯-2-(1H-咪唑-1-基)苯胺(469.0毫克，2.42毫摩尔)和吡啶(382.0毫克，4.84毫摩尔)的二氯甲烷(10.0毫升)中滴加2-氯-2-氧代乙酸甲酯(354.0毫克，2.91毫摩尔)，滴毕，室温反应30分钟。 Under N 2 protection and ice bath, add 5-chloro-2-(1H-imidazol-1-yl)aniline (469.0 mg, 2.42 mmol) and pyridine (382.0 mg, 4.84 mmol) to dichloromethane (10.0 Methyl 2-chloro-2-oxoacetate (354.0 mg, 2.91 mmol) was added dropwise to the solution (ml). After the drop 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(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×3次)洗涤，然后用无水硫酸钠干燥，最后硅胶柱层析纯化(洗脱剂：正己烷：乙酸乙酯＝1：1)得到594.0毫克白色固体2-((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酸甲酯(收率：79.3％)。LCMS:RT＝3.97min,[M+H] +＝280.33。 After the reaction was over, it was quenched by adding water, and the mixture was extracted with dichloromethane (50 ml×3 times). The organic phases were combined, and the organic phase was washed with saturated brine (30 ml×3 times), then dried over anhydrous sodium sulfate, and finally purified by silica gel column chromatography (eluent: n-hexane: ethyl acetate = 1:1) 594.0 mg of methyl 2-((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetate was obtained as a white solid (yield: 79.3%). LCMS: RT = 3.97 min, [M+H] + =280.33.
步骤D：合成2-((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酸Step D: Synthesis of 2-((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000564
Figure PCTCN2020133493-appb-000564
冰浴下，向含有2-((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酸甲酯(594.0毫克，2.12毫摩尔)的四氢呋喃(28.0毫升)中滴加氢氧化锂(179.0毫克，4.25毫摩尔)水溶液(14.0毫升)，滴毕即反应结束。Under an ice bath, add methyl 2-((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetate (594.0 mg, 2.12 mmol) in tetrahydrofuran (28.0 Lithium hydroxide (179.0 mg, 4.25 mmol) aqueous solution (14.0 ml) was added dropwise to the solution (14.0 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(30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，最后减压浓缩得到538毫克淡黄色固体合成2-((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酸，无需纯化，直接用于下一步反应(收率：95.5％)。LCMS:RT＝4.26min,[M-H] -＝264.11。 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 (30 ml×3 times), combine the organic phases, and use saturated brine (30 ML×2 times), then dried with anhydrous sodium sulfate, and finally concentrated under reduced pressure to obtain 538 mg of pale yellow solid synthetic 2-((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino) -2-oxoacetic acid, without purification, was directly used in the next reaction (yield: 95.5%). LCMS: RT = 4.26 min, [MH] - =264.11.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000565
Figure PCTCN2020133493-appb-000565
室温下，将2-((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酸(25.0毫克，0.10毫摩尔)、(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(38.0毫克，0.11毫摩尔)和1-丙基磷酸酐(120.0毫克，0.38毫摩尔)加入N,N-二甲基甲酰胺(2.0毫升)中，滴加N,N-二异丙基乙胺(36.4毫克，0.30毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 2-((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetic acid (25.0 mg, 0.10 mmol), (S)-4-( 2-Amino-3-phenylpropionamido) tert-butyl benzoate (38.0 mg, 0.11 mmol) and 1-propyl phosphoric anhydride (120.0 mg, 0.38 mmol) were added to N,N-dimethylformamide (2.0 ml), N,N-diisopropylethylamine (36.4 mg, 0.30 mmol) was added dropwise, after the dropping, the reaction was carried out at room temperature for 5 hours under the protection of N 2.
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＝5/1)。得到93毫克黄色固体(S)-4-(2-(2-(((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：37.2％)。LCMS:RT＝4.37min,[M-H] -＝586.23。 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=5/1). Obtain 93 mg of yellow solid (S)-4-(2-(2-(((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-phenylpropionamido)benzoate (yield: 37.2%). LCMS: RT = 4.37 min, [MH] - =586.23.
步骤F:合成(S)-4-(2-(2-((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step F: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetamido)-3- Phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000566
Figure PCTCN2020133493-appb-000566
室温下，将(S)-4-(2-(2-(((5-氯-2-(1H-咪唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(24.0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2小时。At room temperature, the (S)-4-(2-(2-(((5-chloro-2-(1H-imidazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido) tert-butyl benzoate (24.0 mg, 0.04 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 2 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23.6毫克白色固体合成(S)-4-(2-(2-(((5-氯-2-环丁氧基苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：97.6％)。LCMS:RT＝3.21min,[M+H] +＝532.15。 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 23.6 mg of white solid synthetic (S)-4-(2-(2-(((5-chloro-2-cyclobutoxyphenyl)amino)-2-oxoacetamide (Yl)-3-phenylpropionamido)benzoic acid (yield: 97.6%). LCMS: RT = 3.21 min, [M+H] + = 532.15.
实施例103Example 103
合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-甲氧基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-methoxy Propyl propionamido) benzoic acid
Figure PCTCN2020133493-appb-000567
Figure PCTCN2020133493-appb-000567
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-4-(2-(((叔丁氧基羰基)氨基)-3-甲氧基丙酰胺基)苯甲酸叔丁酯Step A: Synthesis of tert-butyl (S)-4-(2-(((tert-butoxycarbonyl)amino)-3-methoxypropionamido)benzoate
Figure PCTCN2020133493-appb-000568
Figure PCTCN2020133493-appb-000568
室温下，将N-(叔丁氧羰基)-O-甲基-L-丝氨酸(1.0克，4.56毫摩尔)、4-氨基苯甲酸叔丁酯(801.0毫克，4.15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(2.4克，6.23毫摩尔)加入N,N-二甲基甲酰胺(2.0毫升)中，滴加N,N-二异丙基乙胺(1.1毫克，8.30毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, N-(tert-butoxycarbonyl)-O-methyl-L-serine (1.0 g, 4.56 mmol), tert-butyl 4-aminobenzoate (801.0 mg, 4.15 mmol) and 2-( 7-oxybenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.4g, 6.23mmol) was added to N,N-dimethylformamide (2.0ml) In the middle, N,N-diisopropylethylamine (1.1 mg, 8.30 mmol) was added dropwise, and after the dropping, the reaction was carried out at room temperature for 5 hours under the protection of N 2.
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＝5/1)。得到1.4克黄色固体(S)-4-(2-(((叔丁氧基羰基)氨基)-3-甲氧基丙酰胺基)苯甲酸叔丁酯(收率：85.8％)。LCMS:RT＝4.23min,[M-H] -＝393.26。 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=5/1). Yield 1.4 g of tert-butyl (S)-4-(2-(((tert-butoxycarbonyl)amino)-3-methoxypropionamido)benzoate as a yellow solid (yield: 85.8%). LCMS: RT = 4.23 min, [MH] - =393.26.
步骤B：合成(S)-4-(2-氨基-3-甲氧基丙酰胺基)苯甲酸叔丁酯Step B: Synthesis of tert-butyl (S)-4-(2-amino-3-methoxypropionamido)benzoate
Figure PCTCN2020133493-appb-000569
Figure PCTCN2020133493-appb-000569
冰浴条件下，将(S)-4-(2-(((叔丁氧基羰基)氨基)-3-甲氧基丙酰胺基)苯甲酸叔丁酯(800.0毫克，2.03毫摩尔)、4-氨基苯甲酸叔丁酯(801.0毫克，4.15毫摩尔)加入盐酸乙酸乙酯混合液(盐酸2.0毫升，乙酸乙酯8.0毫升)中，N 2保护下，室温反应5小时。 Under ice bath conditions, (S)-4-(2-(((tert-butoxycarbonyl)amino)-3-methoxypropionamido) tert-butyl benzoate (800.0 mg, 2.03 mmol), Tert-Butyl 4-aminobenzoate (801.0 mg, 4.15 mmol) was added to the mixed solution of hydrochloric acid and ethyl acetate (2.0 ml of hydrochloric acid, 8.0 ml of ethyl acetate), and the reaction was carried out at room temperature for 5 hours under the protection of N 2.
反应结束，混合液中加乙酸乙酯(50毫升)，冰浴下调pH至8.0，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＝10/1)。得到356.0毫克黄色固体(S)-4-(2-氨基-3-甲氧基丙酰胺基)苯甲酸叔丁酯(收率：86.1％)。LCMS:RT＝4.39min,[M-H] -＝293.31。 After the reaction is over, add ethyl acetate (50 ml) to the mixed solution, adjust the pH to 8.0 on an ice bath, extract, combine the organic phases, wash the organic phases with saturated brine (20 ml × 2 times), and then with anhydrous sodium sulfate Dry, and finally concentrate under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: ethyl acetate/n-hexane=10/1). Obtained 356.0 mg of tert-butyl (S)-4-(2-amino-3-methoxypropionamido)benzoate as a yellow solid (yield: 86.1%). LCMS: RT = 4.39 min, [MH] - =293.31.
步骤C：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-甲氧基丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Methoxypropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000570
Figure PCTCN2020133493-appb-000570
室温下，将(S)-4-(2-氨基-3-甲氧基丙酰胺基)苯甲酸叔丁酯(604.0毫克，2.05毫摩尔)、2-((5-氯-2-(1H-四唑-1-基)苯 基)氨基)-2-氧乙酸(659.0毫克，2.46毫摩尔)和1-丙基磷酸酐(2.6克，8.20毫摩尔)加入N,N-二甲基甲酰胺(15.0毫升)中，滴加N,N-二异丙基乙胺(793.0毫克，6.15毫摩尔)，滴毕，N 2保护下，室温反应5小时。 At room temperature, (S)-4-(2-amino-3-methoxypropionamido) tert-butyl benzoate (604.0 mg, 2.05 mmol), 2-((5-chloro-2-(1H -Tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (659.0 mg, 2.46 mmol) and 1-propyl phosphoric anhydride (2.6 g, 8.20 mmol) were added to N,N-dimethylformaldehyde To the amide (15.0 ml), N,N-diisopropylethylamine (793.0 mg, 6.15 mmol) was added dropwise, after the dropping, the reaction was carried out at room temperature for 5 hours under the protection of N 2.
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＝2/1)。得到113.0毫克黄色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-甲氧基丙酰胺基)苯甲酸叔丁酯(收率：10.1％)。LCMS:RT＝4.41min,[M-H] -＝542.28。 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=2/1). 113.0 mg of yellow solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- Tert-Butyl 3-methoxypropionamido)benzoate (yield: 10.1%). LCMS: RT = 4.41 min, [MH] - =542.28.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-甲氧基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Methoxypropionamido) benzoic acid
Figure PCTCN2020133493-appb-000571
Figure PCTCN2020133493-appb-000571
室温下，将(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-甲氧基丙酰胺基)苯甲酸叔丁酯(106.0毫克，0.20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2小时。At room temperature, the (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Methoxypropionamido) tert-butyl benzoate (106.0 mg, 0.20 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 2 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12.7毫克白色固体合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-甲氧基丙酰胺基)苯甲酸(收率：93.1％)。LCMS:RT＝3.34min,[M-H] -＝486.12。 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 12.7 mg of white solid synthetic (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido)-3-methoxypropionamido)benzoic acid (yield: 93.1%). LCMS: RT = 3.34 min, [MH] - = 486.12.
实施例104Example 104
合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((S)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((S)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000572
Figure PCTCN2020133493-appb-000572
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((S)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((S)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000573
Figure PCTCN2020133493-appb-000573
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(225.0毫克，0.30毫摩尔)、(S)-N,N-二甲基吡咯烷-3-胺盐酸盐(165.0毫克，0.90毫摩尔)溶于四氢呋喃中，滴加N,N-二异丙基乙胺(304.0毫克，2.36毫摩尔)，滴毕，N 2保护下，60摄氏度过夜反应。 At room temperature, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (225.0 mg, 0.30 mmol), (S)-N,N-di Methylpyrrolidin-3-amine hydrochloride (165.0 mg, 0.90 mmol) was dissolved in tetrahydrofuran, and N,N-diisopropylethylamine (304.0 mg, 2.36 mmol) was added dropwise. After the drop, N 2 Under protection, react overnight at 60 degrees Celsius.
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＝2/1)。得到204.0毫克黄色固体5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(S)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：53.0％)。LCMS:RT＝4.09min,[M+H] +＝784.06。 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=2/1). Obtain 204.0 mg of yellow solid 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(S)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (Yield: 53.0% ). LCMS: RT = 4.09 min, [M+H] + =784.06.
步骤B：合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((S)-3(S)-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((S)-3(S)-(dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000574
Figure PCTCN2020133493-appb-000574
室温下，将5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((S)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(204.0毫克，0.26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2小时。At room temperature, the 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((S)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (204.0 mg, 0.26 mg Mol) was added to methylene chloride (5.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 2 hours.
反应结束，蒸干二氯甲烷并用油泵拉干三氟乙酸，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35.2毫克白色固体5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((S)-3(S)-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：93.7％)。LCMS:RT＝3.31min,[M-H] -＝726.15。 When the reaction is over, the dichloromethane is evaporated and the trifluoroacetic acid is pulled dry with 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 35.2 mg of white solid 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-((S)-3(S)-(dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2 -Carboxylic acid (yield: 93.7%). LCMS: RT = 3.31 min, [MH] - =726.15.
实施例105Example 105
合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((R)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((R)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000575
Figure PCTCN2020133493-appb-000575
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((R)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((R)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000576
Figure PCTCN2020133493-appb-000576
室温下，将5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((R)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(127.0毫克，0.17毫摩尔)、叔丁基5-((S)-2-(2-((5-氯-2-(1H-(R)-N,N-二甲基吡咯烷-3-胺(57.0毫克，0.50毫摩尔)溶于四氢呋喃(3.0毫升)中，滴加N,N-二异丙基乙胺(98.0毫克，0.91毫摩尔)，N 2保护下，60摄氏度反应5小时。 At room temperature, the 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((R)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (127.0 mg, 0.17 mg Mol), tert-butyl 5-((S)-2-(2-((5-chloro-2-(1H-(R)-N,N-dimethylpyrrolidine-3-amine (57.0 mg, 0.50 mmol) was dissolved in tetrahydrofuran (3.0 ml), and N,N-diisopropylethylamine (98.0 mg, 0.91 mmol) was added dropwise, and the reaction was carried out at 60 degrees Celsius for 5 hours under the protection of N 2.
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133.0毫克黄色固体5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((R)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：40.9％)。LCMS:RT＝4.16min,[M+H] +＝784.46。 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). 133.0 mg of yellow solid 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((R)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (Yield: 40.9 %). LCMS: RT = 4.16 min, [M+H] + =784.46.
步骤B：合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((R)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((R)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000577
Figure PCTCN2020133493-appb-000577
室温下，将5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((R)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(133.0毫克，0.02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2小时。At room temperature, the 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((R)-3-(Dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (133.0 mg, 0.02 mg Mol) was added to methylene chloride (5.0 mL), trifluoroacetic acid (1.0 mL) was added dropwise, and the reaction was carried out at room temperature for 2 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18.2毫克白色固体5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((R)-3-(二甲氨基)吡咯烷-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：93.2％)。LCMS:RT＝3.85min,[M-H] -＝726.18。 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 18.2 mg of white solid 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-((R)-3-(dimethylamino)pyrrolidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid (Yield: 93.2%). LCMS: RT = 3.85 min, [MH] - =726.18.
实施例106Example 106
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-羟基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-Hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000578
Figure PCTCN2020133493-appb-000578
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 PCTCN2020133493-appb-000579
Figure PCTCN2020133493-appb-000579
室温下，将4-羟基哌啶-1-羧酸叔丁酯(5.0克，24.8毫摩尔)加入二氯甲烷(20.0毫升)中，滴加三氟乙酸(10.0毫升)，滴毕，N 2保护下，室温反应5小时。 At room temperature, 4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (5.0 g, 24.8 mmol) was added to dichloromethane (20.0 ml), trifluoroacetic acid (10.0 ml) was added dropwise, and after the drop, N 2 Under protection, react at room temperature for 5 hours.
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＝2/1)，得到1.8克黄色固体哌啶-4-醇的三氟乙酸盐(收率：35.9％)。LCMS:RT＝4.21min,[M-H] -＝198.18。 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=2/1) to obtain 1.8 g of trifluoroacetate of piperidin-4-ol as a yellow solid (yield: 35.9%) . LCMS: RT = 4.21 min, [MH] - =198.18.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-羟基哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step B: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Hydroxypiperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000580
Figure PCTCN2020133493-appb-000580
室温下，向含有叔丁基(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸酯(100.0毫克，0.13毫摩尔)和哌啶-4-醇的三氟乙酸盐(86.2毫克，0.40毫摩尔)的四氢呋喃(3.0毫升)溶液中滴加N,N-二异丙基乙胺(76.0毫克，0.60毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains tert-butyl (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylate (100.0 mg, 0.13 mmol) and piperidin-4-ol N,N-diisopropylethylamine (76.0 mg, 0.60 mmol) was added dropwise to a solution of trifluoroacetate (86.2 mg, 0.40 mmol) in tetrahydrofuran (3.0 mL), the dripping was completed, and the reaction was performed 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：乙酸乙酯/正己烷＝1/6)。得到 165.0毫克白色固体叔丁基(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-羟基哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸酯(收率：51.3％)。LCMS:RT＝4.05min,[M+H] +＝771.55。 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 165.0 mg of white solid tert-butyl(S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Group)-3-(4-(4-hydroxypiperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylate (yield: 51.3%). LCMS:RT= 4.05min, [M+H] + =771.55.
步骤C：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-羟基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step C: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000581
Figure PCTCN2020133493-appb-000581
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-羟基哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(165.0毫克，0.20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, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Hydroxypiperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (165.0 mg, 0.20 mmol) was added to dichloromethane (5.0 (Ml), trifluoroacetic acid (1.0 ml) was added dropwise, and reacted 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12.4毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-羟基哌啶-1-羧酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：96.1％)。LCMS:RT＝4.12min,[M-H] -＝713.08。 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 12.4 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(4-hydroxypiperidine-1-carboxamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 96.1%) . LCMS: RT = 4.12 min, [MH] - = 713.08.
实施例107Example 107
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲基哌嗪-1-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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(4-(4-methylpiperazin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000582
Figure PCTCN2020133493-appb-000582
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲基哌嗪-1-基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(4-Methylpiperazin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000583
Figure PCTCN2020133493-appb-000583
室温下，向含有叔丁基(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸酯(100.0毫克，0.13毫摩尔)和1-甲基-4-(哌啶-4-基)哌嗪盐酸盐(86.2毫克，0.40毫摩尔)的四氢呋喃溶液中滴加N,N-二异丙基乙胺(76.8毫克，0.61毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains tert-butyl (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Group)-3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylate (100.0 mg, 0.13 mmol) and 1-methyl-4- (Piperidin-4-yl)piperazine hydrochloride (86.2 mg, 0.40 mmol) in tetrahydrofuran solution was added dropwise N,N-diisopropylethylamine (76.8 mg, 0.61 mmol), dripping finished, 60 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177.0毫克白色固体叔丁基(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲基哌嗪-1-基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸酯(收率：47.3％)。LCMS:RT＝3.98min,[M+H] +＝853.27。 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). Obtained 177.0 mg of white solid tert-butyl(S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-(4-(4-(4-methylpiperazin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylate ( Yield: 47.3%). LCMS: RT = 3.98 min, [M+H] + =853.27.
步骤B：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲基哌嗪-1-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-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(4-methylpiperazin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000584
Figure PCTCN2020133493-appb-000584
室温下，将叔丁基(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲基哌嗪-1-基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸酯(177.0毫克，0.20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, the tert-butyl (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido )-3-(4-(4-(4-Methylpiperazin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylate (177.0 Mg, 0.20 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113毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(4-甲基哌嗪-1-基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：76.1％)。LCMS:RT＝2.73min,[M+H] +＝797.29。 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 113 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido)-3-(4-(4-(4-methylpiperazin-1-yl)piperidine-1-carboxamido)phenylpropionamido)-1H-indole- 2-carboxylic acid (yield: 76.1%). LCMS: RT = 2.73 min, [M+H] + =797.29.
实施例108Example 108
合成(S)-5-(3-(4-(4-羧基哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(3-(4-(4-carboxypiperidine-1-carboxamido)phenyl)-2-(2-((5-chloro-2-(1H-tetrazole-1 -Yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000585
Figure PCTCN2020133493-appb-000585
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-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(3-(4-(4-(tert-butoxycarbonyl)piperidine-1-carboxamido)phenyl)-2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000586
Figure PCTCN2020133493-appb-000586
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(80.0毫克，0.11毫摩尔)和哌啶-4-羧酸叔丁酯盐酸盐(70.2毫克，0.31毫摩尔)的四氢呋喃溶液中滴加N,N-二异丙基乙胺(61.0毫克，0.47毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (80.0 mg, 0.11 mmol) and piperidine-4-carboxylic acid tert N,N-diisopropylethylamine (61.0 mg, 0.47 mmol) was added dropwise to the tetrahydrofuran solution of butyl ester hydrochloride (70.2 mg, 0.31 mmol), 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：乙酸乙酯/正己烷＝1/3)。得到124毫克白色固体(S)-5-(3-(4-(4-(叔丁氧基羰基)哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：53.2％)。LCMS:RT＝4.11min,[M+H] +＝856.02。 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 124 mg of white solid (S)-5-(3-(4-(4-(tert-butoxycarbonyl)piperidine-1-carboxamido)phenyl)-2-(2-((5-chloro -2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield: 53.2%). LCMS: RT = 4.11 min, [M+H] + =856.02.
步骤B：合成(S)-5-(3-(4-(4-(叔丁氧基羰基)哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step B: Synthesis of (S)-5-(3-(4-(4-(tert-butoxycarbonyl)piperidine-1-carboxamido)phenyl)-2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000587
Figure PCTCN2020133493-appb-000587
室温下，将(S)-5-(3-(4-(4-(叔丁氧基羰基)哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(124.0毫克，0.15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-(tert-butoxycarbonyl)piperidine-1-carboxamido)phenyl)-2-(2-((5-chloro- 2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (124.0 mg, 0.15 mmol) was added Trifluoroacetic acid (1.0 mL) was added dropwise to dichloromethane (5.0 mL), 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9.5毫克白色固体叔丁基(S)-5-(3-(4-(4-(叔丁氧基羰基)哌啶-1-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙酰胺基)-1H-吲哚-2-羧酸(收率：97.5％)。LCMS:RT＝3.40min,[M-H] -＝741.17。 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 9.5 mg of white solid tert-butyl(S)-5-(3-(4-(4-(tert-butoxycarbonyl)piperidine-1-carboxamido)phenyl) -2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetylamino)propionamido)-1H-indole-2-carboxylic acid (Yield: 97.5%). LCMS: RT = 3.40 min, [MH] - =741.17.
实施例109Example 109
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenyl Propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000588
Figure PCTCN2020133493-appb-000588
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000589
Figure PCTCN2020133493-appb-000589
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(87.0毫克，0.14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2小时。At room temperature, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (87.0 mg, 0.14 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 2 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15.3毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：80.9％)。LCMS:RT＝3.90min,[M-H] -＝572.02。 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 15.3 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-phenylpropionamido)-1H-indole-2-carboxylic acid (yield: 80.9%). LCMS: RT = 3.90 min, [MH] - = 572.02.
实施例110Example 110
合成(S)-5-(3-(4-(2-氧代-7-氮杂螺[3.5]壬烷-7-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(3-(4-(2-oxo-7-azaspiro[3.5]nonane-7-carboxamido)phenyl)-2-(2-((5-chloro -2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propylamino)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000590
Figure PCTCN2020133493-appb-000590
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-氧代-7-氮杂螺[3.5]壬烷-7-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙胺基)-1H-吲哚-2-羧酸Step A: Synthesis of (S)-5-(3-(4-(2-oxo-7-azaspiro[3.5]nonane-7-carboxamido)phenyl)-2-(2-(( 5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propylamino)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000591
Figure PCTCN2020133493-appb-000591
室温下，将叔丁基(S)-5-(3-(4-(2-氧-7-氮杂螺环[3.5]壬烷-7-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四氮唑-1-基)苯基)氨基)-2-氧乙酰氨基)丙胺基)-1H-吲哚-2-羧酸盐(56.0毫克，0.07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2小时。At room temperature, the tert-butyl (S)-5-(3-(4-(2-oxo-7-azaspiro[3.5]nonane-7-carboxamido)phenyl)-2-(2 -((5-Chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propylamino)-1H-indole-2-carboxylate (56.0 mg, 0.07 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 2 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6.3毫克白色固体(S)-5-(3-(4-(2-氧代-7-氮杂螺[3.5]壬烷-7-羧酰胺基)苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰氨基)丙胺基)-1H-吲哚-2-羧酸(收率：90.4％)。LCMS:RT＝3.44min,[M+H] +＝741.14。 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 6.3 mg of white solid (S)-5-(3-(4-(2-oxo-7-azaspiro[3.5]nonane-7-carboxamido)phenyl )-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)propylamino)-1H-indole-2-carboxy Acid (yield: 90.4%). LCMS: RT = 3.44 min, [M+H] + =741.14.
实施例111Example 111
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰氨基)-3-(4-(4-(2-羟基乙氧基)哌啶-1-甲酰胺)苯基)丙胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4-( 4-(2-Hydroxyethoxy)piperidine-1-carboxamide)phenyl)propylamino)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000592
Figure PCTCN2020133493-appb-000592
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-羟基乙氧基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(2-Hydroxyethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000593
Figure PCTCN2020133493-appb-000593
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(80.0毫克，0.01毫摩尔)和2-(哌啶-4-氧基)乙烷-1-醇(448.2毫克，1.85毫摩尔)的四氢呋喃中滴加N,N-二异丙基乙胺(299.8毫克，2.31毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (80.0 mg, 0.01 mmol) and 2-(piperidine-4- N,N-diisopropylethylamine (299.8 mg, 2.31 mmol) was added dropwise to the tetrahydrofuran of oxy)ethane-1-ol (448.2 mg, 1.85 mmol), 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：乙酸乙酯/正己烷＝1/8)。得到151.2毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-羟基乙氧基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：45.3％)。LCMS:RT＝3.86min,[M+H] +＝815.77。 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=1/8). 151.2 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-hydroxyethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (Yield: 45.3% ). LCMS: RT = 3.86 min, [M+H] + =815.77.
步骤B：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-羟基乙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(4-(2-hydroxyethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000594
Figure PCTCN2020133493-appb-000594
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-羟基乙氧基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(42.0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, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(2-Hydroxyethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (42.0 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37.0毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(2-羟基乙氧基)哌啶-1-羧酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：92.1％)。LCMS:RT＝3.33min,[M-H] -＝757.18。 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 37.0 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido)-3-(4-(4-(2-hydroxyethoxy)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid (Yield: 92.1%). LCMS: RT = 3.33 min, [MH] - = 757.18.
实施例112Example 112
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((4-甲基苯基)磺酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -((4-Methylphenyl)sulfonamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000595
Figure PCTCN2020133493-appb-000595
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((4-甲基苯基)磺酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((4-Methylphenyl)sulfonamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000596
Figure PCTCN2020133493-appb-000596
室温下，将(S)-5-(3-(4-氨基苯基)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(200.0毫克，0.31毫摩尔)、三乙醇胺(79.0毫克，0.78毫摩尔)、4-甲苯磺酰氯(89.0毫克，0.47毫摩尔)加入二氯甲烷(3.0毫升)中，室温过夜反应。At room temperature, (S)-5-(3-(4-aminophenyl)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido) propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (200.0 mg, 0.31 mmol), triethanolamine (79.0 mg, 0.78 mmol), 4-toluenesulfonyl chloride (89.0 mg, 0.47 mmol) was added to dichloromethane (3.0 mL) and reacted overnight at room temperature.
反应结束，加水淬灭，混合液用二氯甲烷(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：二氯甲烷/乙酸乙酯＝4/1)。得到37.0毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((4-甲基苯基)磺酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯。(收率：9.1％)。LCMS:RT＝4.33min,[M+H] +＝798.19。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with dichloromethane (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 with anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: dichloromethane/ethyl acetate=4/1). Obtained 37.0 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((4-methylphenyl)sulfonamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester. (Yield: 9.1%). LCMS:RT= 4.33 min, [M+H] + =798.19.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((4-甲基苯基)磺酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-((4-methylphenyl)sulfonamido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000597
Figure PCTCN2020133493-appb-000597
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-甲基苯基)磺酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(37.0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, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-Methylphenyl)sulfonamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (37.0 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7.7毫克白色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((4-甲基苯基)磺酰胺基)苯基)丙酰胺基)-1H-吲哚-2-羧酸(收率：97.0％)。LCMS:RT＝3.77min,[M-H] -＝740.06。 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 7.7 mg of white solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-((4-methylphenyl)sulfonamido)phenyl)propionamido)-1H-indole-2-carboxylic acid (yield: 97.0%) . LCMS: RT = 3.77 min, [MH] - =740.06.
实施例113Example 113
合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(4-羟基-2-甲基丁烷-2-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-( 4-(3-(4-hydroxy-2-methylbutane-2-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000598
Figure PCTCN2020133493-appb-000598
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(4-羟基-2-甲基丁烷-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-(4-hydroxy-2-methylbutan-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000599
Figure PCTCN2020133493-appb-000599
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(80.0毫克，0.11毫摩尔)和3-氨基-3-甲基丁-1-醇(32.0毫克，0.31毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(27.0毫克，0.21毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (80.0 mg, 0.11 mmol) and 3-amino-3-methyl N,N-diisopropylethylamine (27.0 mg, 0.21 mmol) was added dropwise to but-1-ol (32.0 mg, 0.31 mmol) in tetrahydrofuran (3.0 ml), 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(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131.0毫克白色固体叔丁基(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(4-羟基-2-甲基丁烷-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸酯(收率：51.0％)。LCMS:RT＝4.24min,[M+H] +＝774.12。 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). Obtained 131.0 mg of white solid tert-butyl(S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamide Yl)-3-(4-(3-(4-hydroxy-2-methylbutane-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylate (closed Rate: 51.0%). LCMS: RT = 4.24 min, [M+H] + =774.12.
步骤B：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(4-羟基-2-甲基丁烷-2-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(4-hydroxy-2-methylbutane-2-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000600
Figure PCTCN2020133493-appb-000600
室温下，将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(4-羟基-2-甲基丁烷-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(131.0毫克，0.17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, the (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-(4-Hydroxy-2-methylbutan-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (131.0 mg, 0.17 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26.4毫克白色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(4-羟基-2-甲基丁烷-2-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：96.6％)。LCMS:RT＝3.44min,[M-H] -＝715.31。 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 26.4 mg of white solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)- 2-oxoacetamido)-3-(4-(3-(4-hydroxy-2-methylbutan-2-yl)ureido)phenylpropionamido)-1H-indole-2- Carboxylic acid (yield: 96.6%). LCMS: RT = 3.44 min, [MH] - =715.31.
实施例114Example 114
合成5-((S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-((2S,3S)-1-羟基-3-甲基戊基-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸Synthesis of 5-((S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(3-((2S,3S)-1-hydroxy-3-methylpentyl-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000601
Figure PCTCN2020133493-appb-000601
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1-羟基-3-甲基戊基-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-(1-hydroxy-3-methylpentyl-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000602
Figure PCTCN2020133493-appb-000602
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(80.0毫克，0.11毫摩尔)和2-氨基-3-甲基戊-1-醇(37.0毫克，0.31毫摩尔)的四氢呋喃(1.0毫升)中滴加N,N-二异丙基乙胺(27毫克，0.20毫摩尔)，滴毕，60摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (80.0 mg, 0.11 mmol) and 2-amino-3-methyl N,N-diisopropylethylamine (27 mg, 0.20 mmol) was added dropwise to pentan-1-ol (37.0 mg, 0.31 mmol) in tetrahydrofuran (1.0 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(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142毫克白色固体5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1-羟基-3-甲基戊基-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：45.0％)。LCMS:RT＝4.05min,[M+H] +＝787.02。 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). 142 mg of white solid 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-(3-(1-hydroxy-3-methylpentyl-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (yield : 45.0%). LCMS: RT = 4.05 min, [M+H] + =787.02.
步骤B：合成5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1-羟基-3-甲基戊基-2-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸Step B: Synthesis of 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-(1-hydroxy-3-methylpentyl-2-yl)ureido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000603
Figure PCTCN2020133493-appb-000603
室温下，将5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1-羟基-3-甲基戊基-2-基)脲基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(142.0毫克，0.18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, the 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(3-(1-hydroxy-3-methylpentyl-2-yl)ureido)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (142.0 mg, 0.18 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142.0毫克白色固体5-((2S)-2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(3-(1-羟基-3-甲基戊基-2-基)脲基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：92.0％)。LCMS:RT＝3.81min,[M-H] -＝730.24。 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 142.0 mg of white solid 5-((2S)-2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2 -Oxoacetamido)-3-(4-(3-(1-hydroxy-3-methylpentyl-2-yl)ureido)phenylpropionamido)-1H-indole-2-carboxy Acid (yield: 92.0%). LCMS: RT = 3.81 min, [MH] - =730.24.
实施例115Example 115
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenyl Propanamido)-1H-benzo[d]imidazole-2-carboxylic acid
Figure PCTCN2020133493-appb-000604
Figure PCTCN2020133493-appb-000604
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000605
Figure PCTCN2020133493-appb-000605
室温下，将(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酰基)-L-苯丙氨酸(19毫克，0.045毫摩尔),5-氨基-1H-苯并[d]咪唑-2-羧酸叔丁酯(13毫克，0.055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(26毫克，0.068毫摩尔)加入N,N-二甲基甲酰胺(2毫升)中，滴加N,N-二异丙基乙胺(18毫克，0.137毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, (2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetyl)-L-phenylalanine (19 mg, 0.045 mg Mol), 5-amino-1H-benzo[d]imidazole-2-carboxylic acid tert-butyl ester (13 mg, 0.055 mmol) and 2-(7-oxybenzotriazole)-N,N,N ',N'-Tetramethylurea hexafluorophosphate (26 mg, 0.068 mmol) was added to N,N-dimethylformamide (2 mL), and N,N-diisopropylethylamine ( 18 mg, 0.137 mmol), after dripping, react overnight at room temperature under N 2 protection.
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＝10/1)。得到7毫克黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸叔丁酯(收率：31.2％)。MS(ESI)M/Z：630.19[M+H] +。 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 = 10/1). Obtain 7 mg of yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-Phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid tert-butyl ester (yield: 31.2%). MS (ESI) M/Z: 630.19 [M+H] + .
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸Step B: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid
Figure PCTCN2020133493-appb-000606
Figure PCTCN2020133493-appb-000606
将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸叔丁酯(27毫克，0.03毫摩尔)溶于二氯甲烷(4毫升)。随后，向上述溶液中加入三氟乙酸(1毫升)。室温反应3小时。(S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenyl Propanamido)-1H-benzo[d]imidazole-2-carboxylic acid tert-butyl ester (27 mg, 0.03 mmol) was dissolved in dichloromethane (4 mL). Subsequently, trifluoroacetic acid (1 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5.4毫克浅黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸(收率：84.7％)。MS(ESI)M/Z：[M+H] +＝574.13。 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, 5.4 mg of light yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxo was obtained. Acetamido)-3-phenylpropionamido)-1H-benzo[d]imidazole-2-carboxylic acid (yield: 84.7%). MS(ESI) M/Z: [M+H] + =574.13.
实施例116Example 116
合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetamido)-3-benzene Propyl propionamido) benzoic acid
Figure PCTCN2020133493-appb-000607
Figure PCTCN2020133493-appb-000607
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成5-氯-2-(四唑-5-基)胺Step A: Synthesis of 5-chloro-2-(tetrazol-5-yl)amine
Figure PCTCN2020133493-appb-000608
Figure PCTCN2020133493-appb-000608
将2-氨基-4-氯苄腈(500毫克，3.28毫摩尔)，三乙胺盐酸盐(588毫克，4.27毫摩尔)，叠氮化钠(277.5毫克，4.27毫摩尔)溶于甲苯(10毫升)中，升温至100度，于氮气氛围下反应过夜。Dissolve 2-amino-4-chlorobenzonitrile (500 mg, 3.28 mmol), triethylamine hydrochloride (588 mg, 4.27 mmol), and sodium azide (277.5 mg, 4.27 mmol) in toluene ( 10 ml), the temperature was raised to 100 degrees, and the reaction was carried out overnight under a nitrogen atmosphere.
将反应液冷却到室温，反应液水洗(10ml×3)，所得水相收集用浓盐酸滴加，有类白色固体析出，过滤，得滤饼600毫克类白色固体5-氯-2-(四唑-5-基)胺(收率：93.6％)。LCMS:RT＝3.23min,[M+H] +＝196.06。 The reaction solution was cooled to room temperature, the reaction solution was washed with water (10ml×3), and the resulting aqueous phase was collected and added dropwise with concentrated hydrochloric acid. An off-white solid precipitated out and filtered to obtain a filter cake of 600 mg off-white solid 5-chloro-2-(four Azol-5-yl)amine (yield: 93.6%). LCMS: RT = 3.23 min, [M+H] + =196.06.
步骤B:合成2-((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧代乙酸甲酯Step B: Synthesis of methyl 2-((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000609
Figure PCTCN2020133493-appb-000609
将5-氯-2-(四唑-5-基)胺(600毫克，3.07毫摩尔)溶于二氯甲烷(10毫升，)中，加入吡啶(368毫克，4.6毫摩尔)，2-氯-2-氧代乙酸甲酯(486毫克，3.99毫摩尔)，室温反应10分钟5-Chloro-2-(tetrazol-5-yl)amine (600 mg, 3.07 mmol) was dissolved in dichloromethane (10 mL,), pyridine (368 mg, 4.6 mmol), 2-chloro Methyl-2-oxoacetate (486 mg, 3.99 mmol), react at room temperature for 10 minutes
加水淬灭反应，旋掉二氯甲烷，乙酸乙酯溶解(30毫升)饱和氯化铵溶液洗(10毫升×3次)，无水硫酸钠干燥得650毫克类白色固体2-((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧代乙酸甲酯(收率：75.2％)。LCMS:RT＝3.32min,[M+H] +＝282.06。 Add water to quench the reaction, spin off the dichloromethane, dissolve in ethyl acetate (30 mL) and wash with saturated ammonium chloride solution (10 mL × 3 times), and dry with anhydrous sodium sulfate to obtain 650 mg of off-white solid 2-((5- Methyl chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetate (yield: 75.2%). LCMS: RT = 3.32 min, [M+H] + =282.06.
步骤C：合成2-((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧乙酸Step C: Synthesis of 2-((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000610
Figure PCTCN2020133493-appb-000610
冰浴下，向含有2-((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧代乙酸甲酯(650毫克，2.3毫摩尔)的四氢呋喃(10毫升)中滴加氢氧化锂(185毫克，4.6毫摩尔)水溶液(2.5毫升)，滴毕反应一小时结束。Under ice bath, add methyl 2-((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetate (650 mg, 2.3 mmol) in tetrahydrofuran ( Lithium hydroxide (185 mg, 4.6 mmol) aqueous solution (2.5 ml) was added dropwise to 10 ml), and the reaction was completed in one hour after dropping.
反应结束，加乙腈有类白色固体析出，过滤，得滤饼600毫克白色固体2-((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧乙酸(收率：97.2％)。LCMS:RT＝2.50min,[M-H] -＝266.02。 After the reaction was over, a white solid precipitated out by adding acetonitrile, and filtered to obtain 600 mg of white solid 2-((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetic acid. (Yield: 97.2%). LCMS: RT = 2.50 min, [MH] - = 266.02.
步骤D：合成(S)-4-(2-(2-((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetamido)-3 -Phenylpropionamido) tert-butyl benzoate
Figure PCTCN2020133493-appb-000611
Figure PCTCN2020133493-appb-000611
室温下，将2-((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧乙酸(300毫克，1.12毫摩尔),(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(400毫克，1.18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(638毫克，1.68毫摩尔)加入N,N-二甲基甲酰胺(10毫升)中，滴加N,N-二异丙基乙胺(433毫克，3.36毫摩尔)，滴毕，N 2保护下，室温反应3小时。 At room temperature, 2-((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetic acid (300 mg, 1.12 mmol), (S)-4-( 2-Amino-3-phenylpropionamido) tert-butyl benzoate (400 mg, 1.18 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetra Methylurea hexafluorophosphate (638 mg, 1.68 mmol) was added to N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (433 mg, 3.36 mmol) was added dropwise ), after dropping, under N 2 protection, react at room temperature for 3 hours.
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(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260毫克黄色固体(S)-4-(2-(2-((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：39.3％)。LCMS:RT＝4.24min,[M+H] +＝590.21。 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 (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 260 mg of yellow solid (S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetamido)- Tert-butyl 3-phenylpropionamido)benzoate (yield: 39.3%). LCMS: RT = 4.24 min, [M+H] + =590.21.
步骤E：合成(S)-4-(2-(2-(((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step E: Synthesis of (S)-4-(2-(2-(((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetamido)- 3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000612
Figure PCTCN2020133493-appb-000612
将(S)-4-(2-(2-((5-氯-2-(1H-四唑-5-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(260毫克，0.44毫摩尔)溶于二氯甲烷(4毫升)。随后，向上述溶液中加入三氟乙酸(1毫升)。室温反应3小时。(S)-4-(2-(2-((5-chloro-2-(1H-tetrazol-5-yl)phenyl)amino)-2-oxoacetamido)-3-phenyl Tert-butyl propionamido)benzoate (260 mg, 0.44 mmol) was dissolved in dichloromethane (4 mL). Subsequently, trifluoroacetic acid (1 mL) was added to the above solution. React at room temperature for 3 hours.
于反应液中加入大量无水乙醚放冰箱冷冻30分钟，有浅红色固体析出，过滤得滤饼5.45毫克浅红色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)-1H-苯并[d]咪唑-2-羧酸(收率：85.5％)。LCMS:RT＝3.63min,[M+H] +＝534.19。 A large amount of anhydrous ether was added to the reaction solution and placed in the refrigerator for 30 minutes. A light red solid precipitated out. The filter cake was filtered to obtain 5.45 mg of a light red solid (S)-5-(2-(2-((5-chloro-2- (1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)-1H-benzo(d)imidazole-2-carboxylic acid (yield : 85.5%). LCMS: RT=3.63min, [M+H] + =534.19.
实施例117Example 117
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(Dimethylamino)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000613
Figure PCTCN2020133493-appb-000613
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯Step A: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Dimethylamino)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester
Figure PCTCN2020133493-appb-000614
Figure PCTCN2020133493-appb-000614
室温下，向含有(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-((苯氧羰基)氨基)苯基)丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(100毫克，0.13毫摩尔)和N，N-二甲基哌啶-4-胺(105毫克，0.52毫摩尔)的四氢呋喃(3.0毫升)中滴加N,N-二异丙基乙胺(168毫克，1.3毫摩尔)，滴毕，65摄氏度反应过夜。At room temperature, it contains (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 3-(4-((phenoxycarbonyl)amino)phenyl)propionamido)-1H-indole-2-carboxylic acid tert-butyl ester (100mg, 0.13mmol) and N,N-dimethylpiperidine N,N-diisopropylethylamine (168 mg, 1.3 mmol) was added dropwise to pyridine-4-amine (105 mg, 0.52 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90毫克淡黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(收率：86.2％)。LCMS:RT＝4.23min,[M+H] +＝798.28。 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 90 mg of light yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -3-(4-(4-(Dimethylamino)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (Yield: 86.2%) . LCMS: RT = 4.23 min, [M+H] + =798.28.
步骤B：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基)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-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3 -(4-(4-(Dimethylamino)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000615
Figure PCTCN2020133493-appb-000615
将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸叔丁酯(90毫克，0.11毫摩尔)溶于二氯甲烷(10毫升)。随后，向上述溶液中加入三氟乙酸(2.0毫升)。室温反应3小时。Add (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-3-(4 -(4-(Dimethylamino)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid tert-butyl ester (90 mg, 0.11 mmol) dissolved in dichloride Methane (10 mL). Then, trifluoroacetic acid (2.0 mL) was added to the above solution and reacted 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51毫克浅黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-3-(4-(4-(二甲基氨基)哌啶-1-甲酰胺基)苯基丙酰胺基)-1H-吲哚-2-羧酸(收率：61.0％)。LCMS:RT＝2.83min,[M+H] +＝742.25。 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, 51 mg of light yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxo was obtained. Acetamido)-3-(4-(4-(dimethylamino)piperidine-1-carboxamido)phenylpropionamido)-1H-indole-2-carboxylic acid (yield: 61.0% ). LCMS: RT = 2.83 min, [M+H] + =742.25.
实施例118Example 118
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-4-(4-甲基哌嗪-1-基)-4-氧杂戊酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-4-(4 -Methylpiperazin-1-yl)-4-oxavaleramido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000616
Figure PCTCN2020133493-appb-000616
步骤A：合成(S)-2-(((叔丁氧羰基)氨基)甲基-4-(4-甲基哌嗪-1-基)-4-氧代丁酸甲酯Step A: Synthesis of methyl (S)-2-(((tert-butoxycarbonyl)amino)methyl-4-(4-methylpiperazin-1-yl)-4-oxobutanoate
Figure PCTCN2020133493-appb-000617
Figure PCTCN2020133493-appb-000617
室温下，将(S)-3-((叔丁氧羰基)氨基)-4-甲氧基-4-氧代丁酸(620毫克，2.5毫摩尔),1-甲基哌嗪(275毫克，2.75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(1.43克，3.76毫摩尔)加入N,N-二甲基甲酰胺(10毫升)中，滴加N,N-二异丙基乙胺(967毫克，7.5毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, (S)-3-((tert-butoxycarbonyl)amino)-4-methoxy-4-oxobutanoic acid (620 mg, 2.5 mmol), 1-methylpiperazine (275 mg , 2.75 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.43 g, 3.76 mmol) was added to N,N- In dimethylformamide (10 ml), N,N-diisopropylethylamine (967 mg, 7.5 mmol) was added dropwise, and 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(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＝1/1)。得到650毫克黄色固体(S)-2-(((叔丁氧羰基)氨基)甲基-4-(4-甲基哌嗪-1-基)-4-氧代丁酸甲酯(收率：78.8％)。LCMS:RT＝1.37min,[M+H] +＝330.18。 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 (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 650 mg of yellow solid (S)-2-(((tert-butoxycarbonyl)amino)methyl-4-(4-methylpiperazin-1-yl)-4-oxobutanoate (yield : 78.8%). LCMS: RT=1.37 min, [M+H] + =330.18.
步骤B：合成(S)-2-((叔丁氧羰基)氨基)-4-(4-甲基哌嗪-1-基)-4-氧代丁酸Step B: Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-4-(4-methylpiperazin-1-yl)-4-oxobutanoic acid
Figure PCTCN2020133493-appb-000618
Figure PCTCN2020133493-appb-000618
冰浴下，向含有(S)-2-(((叔丁氧羰基)氨基)甲基-4-(4-甲基哌嗪-1-基)-4-氧代丁酸甲酯(650毫克，1.97毫摩尔)的四氢呋喃(10毫升)中滴加氢氧化锂(162毫克，3.95毫摩尔)水溶液(2.5毫升)，滴毕反应20分钟结束。Under ice bath, add (S)-2-(((tert-butoxycarbonyl)amino)methyl-4-(4-methylpiperazin-1-yl)-4-oxobutanoate (650 (10 mg, 1.97 mmol) of tetrahydrofuran (10 mL) was added dropwise lithium hydroxide (162 mg, 3.95 mmol) aqueous solution (2.5 mL), and the reaction was completed in 20 minutes after dropping.
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(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。最后减压浓缩得到600毫克淡黄色固体(S)-2-((叔丁氧羰基)氨基)-4-(4-甲基哌嗪-1-基)-4-氧代丁酸。LCMS:RT＝0.77min,[M-H] -＝314.10。 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 (30 ml×3 times), combine the organic phases, and use saturated brine (10 (Ml×2 times) washing, then drying with anhydrous sodium sulfate, and finally concentration under reduced pressure. Finally, it was concentrated under reduced pressure to obtain 600 mg of light yellow solid (S)-2-((tert-butoxycarbonyl)amino)-4-(4-methylpiperazin-1-yl)-4-oxobutanoic acid. LCMS: RT=0.77 min, [MH] - =314.10.
步骤C：合成(S)-5-(2-(((叔丁氧基羰基)氨基)-4-(4-甲基哌嗪-1-基)-4-氧杂戊酰胺基)-1H-吲哚-1,2-二羧酸二叔丁基酯Step C: Synthesis of (S)-5-(2-(((tert-butoxycarbonyl)amino)-4-(4-methylpiperazin-1-yl)-4-oxavaleramido)-1H -Indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000619
Figure PCTCN2020133493-appb-000619
室温下，将(S)-2-((叔丁氧羰基)氨基)-4-(4-甲基哌嗪-1-基)-4-氧代丁酸(620毫克，2.5毫摩尔),5-氨基-1H-吲哚-1,2-二羧酸二叔丁酯(275毫克，2.75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(1.43克，3.76毫摩尔)加入N,N-二甲基甲酰胺(10毫升)中，滴加N,N-二异丙基乙胺(967毫克，7.5毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, (S)-2-((tert-butoxycarbonyl)amino)-4-(4-methylpiperazin-1-yl)-4-oxobutanoic acid (620 mg, 2.5 mmol), 5-amino-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (275 mg, 2.75 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N '-Tetramethylurea hexafluorophosphate (1.43g, 3.76mmol) was added to N,N-dimethylformamide (10ml), and N,N-diisopropylethylamine (967mg, 7.5 mmol), after dripping, react 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(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650毫克黄色固体(S)-2-(((叔丁氧羰基)氨基)甲基-4-(4-甲基哌嗪-1-基)-4-氧代丁酸甲酯(收率：78.8％)。LCMS:RT＝3.32min,[M+H] +＝630.28。 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 (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 650 mg of yellow solid (S)-2-(((tert-butoxycarbonyl)amino)methyl-4-(4-methylpiperazin-1-yl)-4-oxobutanoate (yield : 78.8%). LCMS: RT = 3.32 min, [M+H] + =630.28.
步骤D：合成(S)-5-(2-氨基-4-(4-甲基哌嗪-1-基)-4-氧杂烟酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step D: Synthesis of (S)-5-(2-amino-4-(4-methylpiperazin-1-yl)-4-oxanicotinamido)-1H-indole-1,2-dicarboxy Di-tert-butyl ester
Figure PCTCN2020133493-appb-000620
Figure PCTCN2020133493-appb-000620
将(S)-2-(((叔丁氧羰基)氨基)甲基-4-(4-甲基哌嗪-1-基)-4-氧代丁酸甲酯(650毫克，1.03毫摩尔)溶于乙酸乙酯(10毫升)。随后，向上述溶液中加入2摩尔的盐酸乙酸乙酯溶液(2.0毫升)。室温反应3小时。(S)-2-(((tert-butoxycarbonyl)amino)methyl-4-(4-methylpiperazin-1-yl)-4-oxobutanoate (650 mg, 1.03 mmol ) Was dissolved in ethyl acetate (10 ml). Then, 2 moles of hydrochloric acid ethyl acetate solution (2.0 ml) was added to the above solution and reacted at room temperature for 3 hours.
将反应液减压蒸馏旋干得粗品500毫克浅黄色固体(S)-5-(2-氨基-4-(4-甲基哌嗪-1-基)-4-氧杂烟酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：91.0％)。LCMS:RT＝2.75min,[M+H] +＝530.29。 The reaction solution was distilled under reduced pressure and spin-dried to obtain a crude product of 500 mg of light yellow solid (S)-5-(2-amino-4-(4-methylpiperazin-1-yl)-4-oxanicotinamide)- 1H-Indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 91.0%). LCMS: RT = 2.75 min, [M+H] + = 530.29.
步骤E：合成(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-4-(4-甲基哌嗪-1-基)-4-氧杂戊酰胺基)-1H-吲哚-1,2-二羧酸酯二叔丁酯Step E: Synthesis of (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 4-(4-Methylpiperazin-1-yl)-4-oxavaleramido)-1H-indole-1,2-dicarboxylate di-tert-butyl ester
Figure PCTCN2020133493-appb-000621
Figure PCTCN2020133493-appb-000621
室温下，将(S)-5-(2-氨基-4-(4-甲基哌嗪-1-基)-4-氧杂烟酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(500毫克，0.94毫摩尔),2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(277毫克，1.04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(539毫克，1.42毫摩尔)加入N,N-二甲基甲酰胺(8毫升)中，滴加N,N-二异丙基乙胺(366毫克，2.84毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, (S)-5-(2-amino-4-(4-methylpiperazin-1-yl)-4-oxanicotinamido)-1H-indole-1,2-dicarboxy Di-tert-butyl ester (500 mg, 0.94 mmol), 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (277 mg, 1.04 mmol) Mol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (539 mg, 1.42 mmol) was added to N,N-dimethyl In formamide (8 ml), N,N-diisopropylethylamine (366 mg, 2.84 mmol) was added dropwise, and 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(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：甲醇/二氯甲烷＝10/1)。得到300毫克黄色固体(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-4-(4-甲基哌嗪-1-基)-4-氧杂戊酰胺基)-1H-吲哚-1,2-二羧酸酯二叔丁酯(收率：40.8％)。LCMS:RT＝3.35min,[M+H] +＝779.29。 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 (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 = 10/1). Obtain 300 mg of yellow solid (S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido) -4-(4-Methylpiperazin-1-yl)-4-oxavaleramido)-1H-indole-1,2-dicarboxylate di-tert-butyl ester (yield: 40.8%). LCMS: RT = 3.35 min, [M+H] + =779.29.
步骤F：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-4-(4-甲基哌嗪-1-基)-4-氧杂戊酰胺基)-1H-吲哚-2-羧酸Step F: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-4 -(4-Methylpiperazin-1-yl)-4-oxavaleramido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000622
Figure PCTCN2020133493-appb-000622
将(S)-5-(2-(2-(((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-4-(4-甲基哌嗪-1-基)-4-氧杂戊酰胺基)-1H-吲哚-1,2-二羧酸酯二叔丁酯(90毫克，0.11毫摩尔)溶于二氯甲烷(10毫升)。随后，向上述溶液中加入三氟乙酸(2.0毫升)。室温反应3小时。(S)-5-(2-(2-(((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-4-( 4-Methylpiperazin-1-yl)-4-oxavaleramido)-1H-indole-1,2-dicarboxylate di-tert-butyl ester (90 mg, 0.11 mmol) dissolved in dichloride Methane (10 mL). Then, trifluoroacetic acid (2.0 mL) was added to the above solution and reacted 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25毫克浅黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-4-(4-甲基哌嗪-1-基)-4-氧杂戊酰胺基)-1H-吲哚-2-羧酸(收率：34.7％)。LCMS:RT＝2.59min,[M+H] +＝623.15。 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, 25 mg of light yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxo was obtained. Acetamido)-4-(4-methylpiperazin-1-yl)-4-oxavaleramido)-1H-indole-2-carboxylic acid (yield: 34.7%). LCMS: RT = 2.59 min, [M+H] + =623.15.
实施例119Example 119
合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-5-(4-甲基哌嗪-1-基)-5-氧代戊酰胺基)-1H-吲哚-2-羧酸Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-5-(4 -Methylpiperazin-1-yl)-5-oxopentanamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000623
Figure PCTCN2020133493-appb-000623
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成(S)-2-((叔丁氧羰基)氨基)-5-甲氧基-5-氧戊酸Step A: Synthesis of (S)-2-((tert-butoxycarbonyl)amino)-5-methoxy-5-oxovaleric acid
Figure PCTCN2020133493-appb-000624
Figure PCTCN2020133493-appb-000624
将(S)-2-氨基-5-甲氧基-5-氧戊酸(1.6克，10毫摩尔)溶于二氧六环(10毫升，)中，加入水(10毫升)，三乙胺(3.03克，30毫摩尔)，二碳酸二叔丁酯(3.27克，15毫摩尔)，室温反应三个小时。Dissolve (S)-2-amino-5-methoxy-5-oxovaleric acid (1.6 g, 10 mmol) in dioxane (10 mL), add water (10 mL), triethyl Amine (3.03 g, 30 mmol), di-tert-butyl dicarbonate (3.27 g, 15 mmol), react at room temperature for three hours.
向反应液中加入10％柠檬酸调节PH到3，混合液用乙酸乙酯(100毫升×3次)萃取浓缩得1.1克类白色固体(S)-2-((叔丁氧羰基)氨基)-5-甲氧基-5-氧戊酸(收率：42.5％)，LCMS:RT＝3.22min,[M+H] -＝260.06。 10% citric acid was added to the reaction solution to adjust the pH to 3, and the mixed solution was extracted and concentrated with ethyl acetate (100 ml×3 times) to obtain 1.1 g of off-white solid (S)-2-((tert-butoxycarbonyl)amino) -5-Methoxy-5-oxovaleric acid (yield: 42.5%), LCMS: RT=3.22 min, [M+H] - =260.06.
步骤B：合成(S)-5-(2-((叔丁氧羰基)氨基)-5-甲氧基-5-氧戊二酰氨基)-1H-吲哚-1,2-二羧酸二叔丁基酯Step B: Synthesis of (S)-5-(2-((tert-butoxycarbonyl)amino)-5-methoxy-5-oxoglutarylamino)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester
Figure PCTCN2020133493-appb-000625
Figure PCTCN2020133493-appb-000625
室温下，将(S)-2-((叔丁氧羰基)氨基)-5-甲氧基-5-氧戊酸(1.1克，4.2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(3.2克，8.4毫摩尔)加入N,N-二甲基甲酰胺(10毫升)中，滴加N,N-二异丙基乙胺(1.6克，12.6毫摩尔)，滴毕，N 2保护下，室温反应一小时。 At room temperature, (S)-2-((tert-butoxycarbonyl)amino)-5-methoxy-5-oxovaleric acid (1.1 g, 4.2 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 (3.2 g, 8.4 mmol) was added to N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (1.6 g, 12.6 mmol) was added dropwise. After the drop, N 2 Under protection, react at room temperature for one hour.
反应结束，加水淬灭，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1.1克黄色固体(S)-5-(2-((叔丁氧羰基)氨基)-5-甲氧基-5-氧戊二酰氨基)-1H-吲哚-1,2-二羧酸二叔丁基酯(收率：45.5％)。LCMS:RT＝4.56min,[M+H] +＝576.21。 After the reaction was completed, it was quenched by adding water, the mixture was extracted with ethyl acetate (40 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). Yield 1.1 g of yellow solid (S)-5-(2-((tert-butoxycarbonyl)amino)-5-methoxy-5-oxoglutarylamino)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl acid (yield: 45.5%) LCMS: RT = 4.56 min, [M+H] + = 576.21.
步骤C：合成(S)-5-((1,2-双(叔丁氧羰基)-1H-吲哚-5-基)氨基)-4-((叔丁氧羰基)氨基)-5-氧戊酸Step C: Synthesis of (S)-5-((1,2-bis(tert-butoxycarbonyl)-1H-indol-5-yl)amino)-4-((tert-butoxycarbonyl)amino)-5- Oxovaleric acid
Figure PCTCN2020133493-appb-000626
Figure PCTCN2020133493-appb-000626
冰浴下，向含有(S)-5-(2-((叔丁氧羰基)氨基)-5-甲氧基-5-氧戊二酰氨基)-1H-吲哚-1,2-二羧酸二叔丁基酯(1.1克，1.9毫摩尔)的四氢呋喃(10毫升)中滴加氢氧化锂(153毫克，3.83毫摩尔)水溶液(2.5毫升)，滴毕反应30分钟结束。Under ice bath, add (S)-5-(2-((tert-butoxycarbonyl)amino)-5-methoxy-5-oxoglutarylamino)-1H-indole-1,2-di Lithium hydroxide (153 mg, 3.83 mmol) aqueous solution (2.5 mL) was added dropwise to di-tert-butyl carboxylate (1.1 g, 1.9 mmol) in tetrahydrofuran (10 mL), and the reaction was completed in 30 minutes after dropping.
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(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。最后减压浓缩得到600毫克淡黄色固体(S)-5-((1,2-双(叔丁氧羰基)-1H-吲哚-5-基)氨基)-4-((叔丁氧羰基)氨基)-5-氧戊酸(收率：56.1％)。LCMS:RT＝4.32min,[M-H] -＝560.23。 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 (30 ml×3 times), combine the organic phases, and use saturated brine (10 (Ml×2 times) washing, then drying with anhydrous sodium sulfate, and finally concentration under reduced pressure. Finally, it was concentrated under reduced pressure to obtain 600 mg of light yellow solid (S)-5-((1,2-bis(tert-butoxycarbonyl)-1H-indol-5-yl)amino)-4-((tert-butoxycarbonyl) )Amino)-5-oxovaleric acid (yield: 56.1%). LCMS: RT = 4.32 min, [MH] - = 560.23.
步骤D：合成(S)-5-(2-((叔丁氧羰基)氨基)-5-(4-甲基哌嗪-1-基)-5-氧戊二酰氨基)-1H-吲哚-1,2-二羧酸二叔丁基酯Step D: Synthesis of (S)-5-(2-((tert-butoxycarbonyl)amino)-5-(4-methylpiperazin-1-yl)-5-oxoglutarylamino)-1H-indino Dole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000627
Figure PCTCN2020133493-appb-000627
室温下，将(S)-5-((1,2-双(叔丁氧羰基)-1H-吲哚-5-基)氨基)-4-((叔丁氧羰基)氨基)-5-氧戊酸(600毫克，1.07毫摩尔),1-甲基哌嗪(160毫克，1.60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(608克，1.60毫摩尔)加 入N,N-二甲基甲酰胺(10毫升)中，滴加N,N-二异丙基乙胺(412毫克，3.2毫摩尔)，滴毕，N 2保护下，室温反应2小时。 At room temperature, (S)-5-((1,2-bis(tert-butoxycarbonyl)-1H-indol-5-yl)amino)-4-((tert-butoxycarbonyl)amino)-5- Oxovaleric acid (600 mg, 1.07 mmol), 1-methylpiperazine (160 mg, 1.60 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'- Tetramethylurea hexafluorophosphate (608 g, 1.60 mmol) was added to N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (412 mg, 3.2 mmol) was added dropwise Mol), after dropping, under N 2 protection, react at room temperature for 2 hours.
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(10毫升×2次)洗涤，然后用无水硫酸钠干燥，最后减压浓缩得到粗品629毫克(S)-5-(2-((叔丁氧羰基)氨基)-5-(4-甲基哌嗪-1-基)-5-氧戊二酰氨基)-1H-吲哚-1,2-二羧酸二叔丁基酯(收率：91.55％)。LCMS:RT＝3.32min,[M+H] +＝644.36。 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 (10 ml×2 times), and then dried over anhydrous sodium sulfate. Finally, it was concentrated under reduced pressure to obtain 629 mg of crude product (S)-5-(2-((tert-butoxycarbonyl)amino)-5-(4-methylpiperazin-1-yl)-5-oxoglutarylamino) -1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 91.55%). LCMS: RT = 3.32 min, [M+H] + =644.36.
步骤E：合成(S)-5-(2-氨基-5-(4-甲基哌嗪-1-基)-5-氧戊二酰胺)-1H-吲哚-1,2-二羧酸二叔丁基酯Step E: Synthesis of (S)-5-(2-amino-5-(4-methylpiperazin-1-yl)-5-oxoglutaramide)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester
Figure PCTCN2020133493-appb-000628
Figure PCTCN2020133493-appb-000628
将(S)-5-(2-((叔丁氧羰基)氨基)-5-(4-甲基哌嗪-1-基)-5-氧戊二酰氨基)-1H-吲哚-1,2-二羧酸二叔丁基酯(629毫克，0.97毫摩尔)溶于乙酸乙酯(12毫升)。随后，向上述溶液中加入2摩尔的盐酸乙酸乙酯溶液(3.0毫升)。室温反应3小时。(S)-5-(2-((tert-butoxycarbonyl)amino)-5-(4-methylpiperazin-1-yl)-5-oxoglutarylamino)-1H-indole-1 Di-tert-butyl 2-dicarboxylate (629 mg, 0.97 mmol) was dissolved in ethyl acetate (12 mL). Subsequently, a 2 molar hydrochloric acid ethyl acetate solution (3.0 ml) was added to the above solution. React at room temperature for 3 hours.
将反应液减压蒸馏旋干得粗品320毫克浅黄色固体(S)-5-(2-氨基-5-(4-甲基哌嗪-1-基)-5-氧戊二酰胺)-1H-吲哚-1,2-二羧酸二叔丁基酯(收率：60.3％)。LCMS:RT＝2.73min,[M+H] +＝544.29。 The reaction solution was distilled under reduced pressure and spin-dried to obtain 320 mg of crude product as a pale yellow solid (S)-5-(2-amino-5-(4-methylpiperazin-1-yl)-5-oxoglutaramide)-1H -Indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 60.3%). LCMS: RT = 2.73 min, [M+H] + =544.29.
步骤F：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-5-(4-甲基哌嗪-1-基)-5-氧代戊酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯Step F: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-5 -(4-Methylpiperazin-1-yl)-5-oxopentanamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester
Figure PCTCN2020133493-appb-000629
Figure PCTCN2020133493-appb-000629
室温下，将(S)-5-(2-氨基-5-(4-甲基哌嗪-1-基)-5-氧戊二酰胺)-1H-吲哚-1,2-二羧酸二叔丁基酯(119毫克，0.2毫摩尔),2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧乙酸(53毫克，0.2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(152毫克，0.4毫摩尔)加入N,N-二甲基甲酰胺(3毫升)中，滴加N,N-二异丙基乙胺(27毫克，0.6毫摩尔)，滴毕，N 2保护下，室温反应过夜。 At room temperature, (S)-5-(2-amino-5-(4-methylpiperazin-1-yl)-5-oxoglutarate)-1H-indole-1,2-dicarboxylic acid Di-tert-butyl ester (119 mg, 0.2 mmol), 2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetic acid (53 mg, 0.2 mmol) Mol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (152 mg, 0.4 mmol) added to N,N-dimethyl In the formamide (3 ml), N,N-diisopropylethylamine (27 mg, 0.6 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(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：甲醇/二氯甲烷＝10/1)。得到90毫克黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-5-(4-甲基哌嗪-1-基)-5-氧代戊酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(收率：55.5％)。LCMS:RT＝3.39min,[M+H] +＝793.33。 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 (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 = 10/1). Obtain 90 mg of yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)- 5-(4-Methylpiperazin-1-yl)-5-oxopentanamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (yield: 55.5%). LCMS: RT = 3.39 min, [M+H] + =793.33.
步骤G：合成(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-5-(4-甲基哌嗪-1-基)-5-氧代戊酰胺基)-1H-吲哚-2-羧酸Step G: Synthesis of (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-5 -(4-Methylpiperazin-1-yl)-5-oxopentanamido)-1H-indole-2-carboxylic acid
Figure PCTCN2020133493-appb-000630
Figure PCTCN2020133493-appb-000630
将(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺基)-5-(4-甲基哌嗪-1-基)-5-氧代戊酰胺基)-1H-吲哚-1,2-二羧酸二叔丁酯(27毫克，0.03毫摩尔)溶于二氯甲烷(4毫升)。随后，向上述溶液中加入三氟乙酸(1毫升)。室温反应3小时。(S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxoacetamido)-5-(4 -Methylpiperazin-1-yl)-5-oxopentanamido)-1H-indole-1,2-dicarboxylic acid di-tert-butyl ester (27 mg, 0.03 mmol) dissolved in dichloromethane ( 4ml). Subsequently, trifluoroacetic acid (1 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10毫克浅黄色固体(S)-5-(2-(2-((5-氯-2-(1H-四唑-1-基)苯基)氨基)-2-氧代乙酰胺 基)-5-(4-甲基哌嗪-1-基)-5-氧代戊酰胺基)-1H-吲哚-2-羧酸(收率：46.3％)。LCMS:RT＝2.66min,[M+H] +＝637.17。 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, 10 mg of light yellow solid (S)-5-(2-(2-((5-chloro-2-(1H-tetrazol-1-yl)phenyl)amino)-2-oxo was obtained. Acetamido)-5-(4-methylpiperazin-1-yl)-5-oxopentanamido)-1H-indole-2-carboxylic acid (yield: 46.3%). LCMS: RT = 2.66 min, [M+H] + =637.17.
实施例120Example 120
合成(S)-4-(2-(2-(((2-氯-5-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((2-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000631
Figure PCTCN2020133493-appb-000631
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成2-((2-氯-5-氰基苯基)氨基)-2-氧乙酸甲酯Step A: Synthesis of methyl 2-((2-chloro-5-cyanophenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000632
Figure PCTCN2020133493-appb-000632
将3-氨基-4-氯苄腈(200毫克，1.3毫摩尔)溶于二氯甲烷(5毫升，)中，加入吡啶(135毫克，1.7毫摩尔)，2-氯-2-氧代乙酸甲酯(193毫克，1.5毫摩尔)，室温反应一小时Dissolve 3-amino-4-chlorobenzonitrile (200 mg, 1.3 mmol) in dichloromethane (5 mL), add pyridine (135 mg, 1.7 mmol), 2-chloro-2-oxoacetic acid Methyl ester (193 mg, 1.5 mmol), react at room temperature for one hour
加水淬灭反应，旋掉二氯甲烷，乙酸乙酯溶解(30毫升)饱和氯化铵溶液洗(10毫升×3次)，无水硫酸钠干燥得311毫克类白色固体2-((2-氯-5-氰基苯基)氨基)-2-氧乙酸甲酯(收率：99.3％)。MS(ESI)M/Z：239.01[M+H] +。 Add water to quench the reaction, spin off the dichloromethane, dissolve in ethyl acetate (30 mL) and wash with saturated ammonium chloride solution (10 mL × 3 times), and dry with anhydrous sodium sulfate to obtain 311 mg of off-white solid 2-((2- Chloro-5-cyanophenyl)amino)-2-oxyacetic acid methyl ester (yield: 99.3%). MS (ESI) M/Z: 239.01 [M+H] + .
步骤B：合成2-((2-氯-5-氰基苯基)氨基)-2-氧乙酸Step B: Synthesis of 2-((2-chloro-5-cyanophenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000633
Figure PCTCN2020133493-appb-000633
冰浴下，向含有2-((2-氯-5-氰基苯基)氨基)-2-氧乙酸甲酯(311毫克，1.3毫摩尔)的四氢呋喃(15毫升)中滴加氢氧化锂(109毫克，2.6毫摩尔)水溶液(7.5毫升)，滴毕反应一小时结束。Under an ice bath, lithium hydroxide was added dropwise to tetrahydrofuran (15 ml) containing methyl 2-((2-chloro-5-cyanophenyl)amino)-2-oxoacetate (311 mg, 1.3 mmol) (109 mg, 2.6 mmol) aqueous solution (7.5 mL), the reaction is completed after one hour of dropping.
反应结束，旋走四氢呋喃，于体系中滴加一摩尔稀盐酸调节PH至2，有大量白色固体析出，过滤，得滤饼230毫克白色固体2-((2-氯-5-氰基苯基)氨基)-2-氧乙酸(收率：79％)。MS(ESI)M/Z：222.99[M-H] -。 After the reaction was over, the tetrahydrofuran was spun off, and one mole of dilute hydrochloric acid was added dropwise to the system to adjust the pH to 2. A large amount of white solid precipitated out and filtered to obtain 230 mg of white solid 2-((2-chloro-5-cyanophenyl). )Amino)-2-oxoacetic acid (yield: 79%). MS(ESI) M/Z: 222.99 [MH] - .
步骤C：合成(S)-4-(2-(2-(((2-氯-5-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯Step C: Synthesis of (S)-4-(2-(2-(((2-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) Tert-butyl benzoate
Figure PCTCN2020133493-appb-000634
Figure PCTCN2020133493-appb-000634
室温下，将2-((2-氯-5-氰基苯基)氨基)-2-氧乙酸(70毫克，0.31毫摩尔),(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸叔丁酯(152毫克，0.312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(178毫克，0.46毫摩尔)加入N,N-二甲基甲酰胺(2毫升)中，滴加N,N-二异丙基乙胺(121毫克，0.93毫摩尔)，滴毕，N 2保护下，室温反应3小时。 At room temperature, add 2-((2-chloro-5-cyanophenyl)amino)-2-oxoacetic acid (70 mg, 0.31 mmol), (S)-4-(2-amino-3-phenyl Propionamido) tert-butyl benzoate (152 mg, 0.312 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate ( 178 mg, 0.46 mmol) was added to N,N-dimethylformamide (2 mL), N,N-diisopropylethylamine (121 mg, 0.93 mmol) was added dropwise, and the drop was completed. N 2 protection React at room temperature for 3 hours.
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(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29毫克黄色固体(S)-4-(2-(2-(((2-氯-5-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(收率：17.1％)。MS(ESI)M/Z：547.17[M+H] +。 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 (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 29 mg of yellow solid (S)-4-(2-(2-(((2-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido ) Tert-Butyl benzoate (yield: 17.1%). MS (ESI) M/Z: 547.17 [M+H] + .
步骤D：合成(S)-4-(2-(2-(((2-氯-5-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-(((2-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000635
Figure PCTCN2020133493-appb-000635
将(S)-4-(2-(2-(((2-氯-5-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸叔丁酯(29毫克，0.05毫摩尔)溶于二氯甲烷(4毫升)。随后，向上述溶液中加入三氟乙酸(1毫升)。室温反应3小时。(S)-4-(2-(2-(((2-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) benzoic acid tert Butyl ester (29 mg, 0.05 mmol) was dissolved in dichloromethane (4 mL). Then, trifluoroacetic acid (1 mL) was added to the above solution and reacted 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15毫克浅黄色固体(S)-4-(2-(2-(((2-氯-5-氰基苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：57.6％)。MS(ESI)M/Z：491.1[M+H] +。 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, 15 mg of light yellow solid (S)-4-(2-(2-(((2-chloro-5-cyanophenyl)amino)-2-oxoacetamido)-3- Phenylpropionamido)benzoic acid (yield: 57.6%). MS (ESI) M/Z: 491.1 [M+H] + .
实施例121Example 121
合成(S)-4-(2-(2-(((5-氯-2-(三氟甲基)苯基)氨基)-2-氧乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) benzoic acid
Figure PCTCN2020133493-appb-000636
Figure PCTCN2020133493-appb-000636
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧乙酸酯Step A: Synthesis of 2-((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000637
Figure PCTCN2020133493-appb-000637
将5-氯-2-(三氟甲基)苯胺(200毫克，1.0毫摩尔)溶于二氯甲烷(5毫升，)中，加入吡啶(150毫克，1.2毫摩尔)，2-氯-2-氧代乙酸甲酯(105毫克，1.3毫摩尔)，室温反应一小时Dissolve 5-chloro-2-(trifluoromethyl)aniline (200 mg, 1.0 mmol) in dichloromethane (5 mL), add pyridine (150 mg, 1.2 mmol), 2-chloro-2 -Methyl oxoacetate (105 mg, 1.3 mmol), react at room temperature for one hour
加水淬灭反应，旋掉二氯甲烷，乙酸乙酯溶解(30毫升)饱和氯化铵溶液洗(10毫升×3次)，无水硫酸钠干燥得280毫克类白色固体2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧乙酸酯(收率：97.2％)。MS(ESI)M/Z：282.01[M+H] +。 Add water to quench the reaction, spin off the dichloromethane, dissolve in ethyl acetate (30 ml) and wash with saturated ammonium chloride solution (10 ml × 3 times), and dry with anhydrous sodium sulfate to obtain 280 mg of off-white solid 2-((5- Chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetate (yield: 97.2%). MS (ESI) M/Z: 282.01 [M+H] + .
步骤B：合成2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧乙酸Step B: Synthesis of 2-((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000638
Figure PCTCN2020133493-appb-000638
冰浴下，向含有2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧乙酸酯(280毫克，0.99毫摩尔)的四氢呋喃(10毫升)中滴加氢氧化锂(84毫克，1.9毫摩尔)水溶液(5毫升)，滴毕反应一小时结束。Under an ice bath, add dropwise to tetrahydrofuran (10 ml) containing 2-((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetate (280 mg, 0.99 mmol) An aqueous solution (5 ml) of lithium hydroxide (84 mg, 1.9 mmol) was added, and the reaction was completed for one hour after dripping.
反应结束，旋走四氢呋喃，于体系中滴加一摩尔稀盐酸调节PH至2，有大量白色固体析出，过滤，得滤饼110毫克白色固体2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧乙酸(收率：41％)。MS(ESI)M/Z：265.99[M-H] -。 After the reaction was over, the tetrahydrofuran was spun off, and one mole of dilute hydrochloric acid was added dropwise to the system to adjust the pH to 2. A large amount of white solid precipitated out and filtered to obtain 110 mg of white solid 2-((5-chloro-2-(trifluoromethyl) into the filter cake. (Yl)phenyl)amino)-2-oxoacetic acid (yield: 41%). MS(ESI) M/Z: 265.99 [MH] - .
步骤C：合成(S)-4-(2-(2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯Step C: Synthesis of (S)-4-(2-(2-((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetamido)-3-phenylpropane Amido) methyl benzoate
Figure PCTCN2020133493-appb-000639
Figure PCTCN2020133493-appb-000639
室温下，将2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧乙酸(110毫克，0.41毫摩尔),(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸甲酯(123毫克，0.35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(197毫克，0.52毫摩尔)加入N,N-二甲基甲酰胺(4毫升)中，滴加N,N-二异丙基乙胺(134毫克，1.04毫摩尔)，滴毕，N 2保护下，室温反应3小时。 At room temperature, 2-((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetic acid (110 mg, 0.41 mmol), (S)-4-(2-amino- 3-phenylpropionamido) methyl benzoate (123 mg, 0.35 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluoro Phosphate (197 mg, 0.52 mmol) was added to N,N-dimethylformamide (4 mL), N,N-diisopropylethylamine (134 mg, 1.04 mmol) was added dropwise, and the drop was completed. Under the protection of N 2 , the reaction was carried out at room temperature for 3 hours.
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(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56毫克黄色固体(S)-4-(2-(2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯(收率：24.6％)。MS(ESI)M/Z：548.11[M+H] +。 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 (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 56 mg of yellow solid (S)-4-(2-(2-((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetamido)-3-phenyl Methyl propionamido)benzoate (yield: 24.6%). MS (ESI) M/Z: 548.11 [M+H] + .
步骤D：合成(S)-4-(2-(2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetamido)-3-phenylpropane Amido) benzoic acid
Figure PCTCN2020133493-appb-000640
Figure PCTCN2020133493-appb-000640
冰浴下，向含有(S)-4-(2-(2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯(56毫克，0.1毫摩尔)的四氢呋喃(3毫升)中滴加氢氧化锂(9毫克，0.2毫摩尔)水溶液(1.5毫升)，滴毕反应一小时结束。Under ice bath, add (S)-4-(2-(2-((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetamido)-3-benzene Lithium hydroxide (9 mg, 0.2 mmol) aqueous solution (1.5 mL) was added dropwise to methyl propionamido) benzoate (56 mg, 0.1 mmol) in tetrahydrofuran (3 mL), and the reaction was completed in one hour.
反应结束，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 C1819mm*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15毫克浅黄色固体(S)-4-(2-(2-((5-氯-2-(三氟甲基)苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：27％)。MS(ESI)M/Z：534.10[M+H] +。 After the reaction was completed, 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 C1819mm*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, 15 mg of light yellow solid (S)-4-(2-(2-((5-chloro-2-(trifluoromethyl)phenyl)amino)-2-oxoacetamido) -3-Phenylpropionamido)benzoic acid (yield: 27%). MS (ESI) M/Z: 534.10 [M+H] + .
实施例122Example 122
合成(S)-4-(2-(2-(2-(((4-氯苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Synthesis of (S)-4-(2-(2-(2-(((4-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000641
Figure PCTCN2020133493-appb-000641
具体合成路线如下：The specific synthesis route is as follows:
步骤A：合成2-((4-氯苯基)氨基)-2-氧乙酸甲酯Step A: Synthesis of methyl 2-((4-chlorophenyl)amino)-2-oxoacetate
Figure PCTCN2020133493-appb-000642
Figure PCTCN2020133493-appb-000642
将4-氯苯胺(500毫克，3.92毫摩尔)溶于二氯甲烷(10毫升，)中，加入吡啶(576毫克，4.7毫摩尔)，2-氯-2-氧代乙酸甲酯(403毫克，5.09毫摩尔)，室温反应一小时4-Chloroaniline (500 mg, 3.92 mmol) was dissolved in dichloromethane (10 mL,), pyridine (576 mg, 4.7 mmol), methyl 2-chloro-2-oxoacetate (403 mg , 5.09 mmol), react at room temperature for one hour
加水淬灭反应，旋掉二氯甲烷，乙酸乙酯溶解(30毫升)饱和氯化铵溶液洗(10毫升×3次)，无水硫酸钠干燥得810毫克类白色固体2-((4-氯苯基)氨基)-2-氧乙酸甲酯(收率：96.6％)。MS(ESI)M/Z：214.02[M+H] +。 Add water to quench the reaction, spin off the dichloromethane, dissolve ethyl acetate (30 ml) and wash with saturated ammonium chloride solution (10 ml × 3 times), and dry with anhydrous sodium sulfate to obtain 810 mg of off-white solid 2-((4- (Chlorophenyl)amino)-2-oxyacetic acid methyl ester (yield: 96.6%). MS (ESI) M/Z: 214.02 [M+H] + .
步骤B：合成2-((4-氯苯基)氨基)-2-氧乙酸Step B: Synthesis of 2-((4-chlorophenyl)amino)-2-oxoacetic acid
Figure PCTCN2020133493-appb-000643
Figure PCTCN2020133493-appb-000643
冰浴下，向含2-((4-氯苯基)氨基)-2-氧乙酸甲酯(810毫克，3.79毫摩尔)的四氢呋喃(20毫升)中滴加氢氧化锂(319毫克，7.59毫摩尔)水溶液(10毫升)，滴毕反应一小时结束。Under ice bath, to tetrahydrofuran (20 ml) containing methyl 2-((4-chlorophenyl)amino)-2-oxoacetate (810 mg, 3.79 mmol) was added dropwise lithium hydroxide (319 mg, 7.59 Millimoles) aqueous solution (10 ml), the reaction is completed after dripping for one hour.
反应结束，旋走四氢呋喃，于体系中滴加一摩尔稀盐酸调节PH至2，有大量白色固体析出，过滤，得滤饼757毫克白色固体2-((4-氯苯基)氨基)-2-氧乙酸(收率：99％)。MS(ESI)M/Z：198.01[M-H] -。 After the reaction is over, the tetrahydrofuran is spun off, and one mole of dilute hydrochloric acid is added dropwise to the system to adjust the pH to 2. A large amount of white solid precipitates out, filtered to obtain 757 mg of white solid 2-((4-chlorophenyl)amino)-2 filter cake -Oxyacetic acid (yield: 99%). MS (ESI) M/Z: 198.01 [MH] - .
步骤C：合成(S)-4-(2-(2-(2-(((4-氯苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯Step C: Synthesis of (S)-4-(2-(2-(2-(((4-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid Methyl ester
Figure PCTCN2020133493-appb-000644
Figure PCTCN2020133493-appb-000644
室温下，将2-((4-氯苯基)氨基)-2-氧乙酸(757毫克，3.79毫摩尔),(S)-4-(2-氨基-3-苯基丙酰胺基)苯甲酸甲酯(1.27克，3.79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(2.16克，5.68毫摩尔)加入N,N-二甲基甲酰胺(15毫升)中，滴加N,N-二异丙基乙胺(1.47克，11.37毫摩尔)，滴毕，N 2保护下，室温反应3小时。 At room temperature, 2-((4-chlorophenyl)amino)-2-oxoacetic acid (757 mg, 3.79 mmol), (S)-4-(2-amino-3-phenylpropionamido)benzene Methyl formate (1.27 g, 3.79 mmol) and 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.16 g, 5.68 mmol) ) Was added to N,N-dimethylformamide (15 mL), and N,N-diisopropylethylamine (1.47 g, 11.37 mmol) was added dropwise, after dripping, the reaction was carried out at room temperature for 3 hours under N 2 protection .
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＝1/6)。得到1.2克黄色固体(S)-4-(2-(2-(2-(((4-氯苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯(收率：66.6％)。MS(ESI)M/Z：480.15[M+H] +。 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=1/6). Obtain 1.2 g of yellow solid (S)-4-(2-(2-(2-(((4-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzene Methyl formate (yield: 66.6%). MS (ESI) M/Z: 480.15 [M+H] + .
步骤D：合成(S)-4-(2-(2-(2-(((4-氯苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸Step D: Synthesis of (S)-4-(2-(2-(2-(((4-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido)benzoic acid
Figure PCTCN2020133493-appb-000645
Figure PCTCN2020133493-appb-000645
冰浴下，向含有(S)-4-(2-(2-(2-(((4-氯苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸甲酯(150毫克，0.3毫摩尔)的四氢呋喃(5毫升)中滴加氢氧化钠(50毫克，1.25毫摩尔)水溶液(2.5毫升)，滴毕反应一小时结束。Under an ice bath, to contain (S)-4-(2-(2-(2-(((4-chlorophenyl)amino)-2-oxoacetamido)-3-phenylpropionamido) Sodium hydroxide (50 mg, 1.25 mmol) aqueous solution (2.5 ml) was added dropwise to methyl benzoate (150 mg, 0.3 mmol) in tetrahydrofuran (5 ml), and the reaction was completed in one hour.
反应结束，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 C1819mm*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60毫克浅黄色固体(S)-4-(2-(2-(2-(((4-氯苯基)氨基)-2-氧代乙酰胺基)-3-苯基丙酰胺基)苯甲酸(收率：41.3％)。MS(ESI)M/Z：466.15[M+H] +。 After the reaction was completed, 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 C1819mm*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, 60 mg of light yellow solid (S)-4-(2-(2-(2-(((4-chlorophenyl)amino)-2-oxoacetamido)-3-phenyl Propanamido)benzoic acid (yield: 41.3%). MS (ESI) M/Z: 466.15 [M+H] + .
测试例1：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Test Example 1: 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。 GraphPad Prism software was used to perform curve fitting and calculate the IC 50 value, as shown in Table 1.
表一：本发明化合物对人FXIa抑制的IC50Table 1: The IC50 of the compounds of the present invention for inhibiting human FXIa
Figure PCTCN2020133493-appb-000646
Figure PCTCN2020133493-appb-000646
Figure PCTCN2020133493-appb-000647
Figure PCTCN2020133493-appb-000647
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.
测试例2 本发明化合物对人血浆体外抗凝血作用的测定Test Example 2 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 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 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 PCTCN2020133493-appb-000648
Figure PCTCN2020133493-appb-000648
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.
测试例3 本发明化合物的大鼠药代动力学特征考察Test Example 3 Investigation of the pharmacokinetic characteristics of the compound of the invention in rats
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。SD rats: male, 180-250g, purchased from Guangdong Medical Experimental Animal Center.
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(U300UPLC，TSQ QUANTUMN ULTRA三重四级杆质谱)。Instrument: Thermo Fisher LC-MS (U300UPLC, 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，大鼠静脉给药后，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 the DMSO-PEG-400-physiological saline (5:60:35, v/v/v) system. After intravenous administration to rats, at 5min, 15min, 30min, 1h, 2h, 4h, 6h At 8h and 24h, 200μL of venous blood was collected in a heparinized EP tube, centrifuged at 12000rpm for 2min, and the plasma was frozen at -80℃ 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 and add acetonitrile to dilute to prepare a standard series solution. Accurately draw 20μL of each of the above 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 and 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-18UPLC柱，100*2.1mm，1.9μm。Chromatographic column: Thermo Scientific HYPERSIL GOLD C-18UPLC 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)
Figure PCTCN2020133493-appb-000649
Figure PCTCN2020133493-appb-000649
Figure PCTCN2020133493-appb-000650
Figure PCTCN2020133493-appb-000650
结论：本发明化合物在大鼠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, 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 PCTCN2020133493-appb-100001
    Figure PCTCN2020133493-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、-(CH 2)n-CO-R 5，其中R 4选自-NR 5-(CH 2)n-CO-(CH 2)n-NR 6R 7、
    Figure PCTCN2020133493-appb-100002
    -NR 5-SO 2-NR 6R 7、-NR 5-SO 2-R 13；     R 2 is selected from R 4 substituted or unsubstituted benzene ring, -(CH 2 )n-CO-R 5 , wherein R 4 is selected from -NR 5 -(CH 2 )n-CO-(CH 2 )n-NR 6 R 7 ,
    Figure PCTCN2020133493-appb-100002
    -NR 5 -SO 2 -NR 6 R 7 , -NR 5 -SO 2 -R 13 ;
    Ar选自至少一个R 8取代或者未取代的以下基团：
    Figure PCTCN2020133493-appb-100003
    Figure PCTCN2020133493-appb-100004
    Ar is selected from at least one of the following groups substituted or unsubstituted by R 8:
    Figure PCTCN2020133493-appb-100003
    Figure PCTCN2020133493-appb-100004
    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、苯基、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 PCTCN2020133493-appb-100005
    -(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, phenyl, C 1-4 alkyl, C 1-4 alkoxy-C 1-4 alkyl, -SO 2 -C 1-4 alkane , -SO 2 -benzene, -C 1-6 monohydric or dihydric alcohol,
    Figure PCTCN2020133493-appb-100005
    -(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、-CO-NR 10、-CONR 10R 11、-CONR 10-SO 2-C 1-4烷、氰基、-NR 10R 11、NR 10R 11-C 1-4的烷氧基、-C 1-4的羧酸、C 1-4的烷基、C 1-4的烷氧基、C 1-4的烷氧-C 1-4烷氧基、-CO-吗啉、吗啉、-O-C 1-6的一元或者二元醇、HOOC-C 1-4烷氧基、
    Figure PCTCN2020133493-appb-100006
    R 9 is selected from hydrogen, -(CH 2 )n-OH, -SO 2 -C 1-4 alkane, -CO-NR 10 , -CONR 10 R 11 , -CONR 10 -SO 2 -C 1-4 alkane, Cyano, -NR 10 R 11 , NR 10 R 11 -C 1-4 alkoxy, -C 1-4 carboxylic acid, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 alkoxy-C 1-4 alkoxy, -CO-morpholine, morpholine, -OC 1-6 mono or dihydric alcohol, HOOC-C 1-4 alkoxy,
    Figure PCTCN2020133493-appb-100006
    R 10、R 11、R 12独立选自氢、C 1-4烷基、-(CH 2)n-OH； R 10 , R 11 , and R 12 are independently selected from hydrogen, C 1-4 alkyl, -(CH 2 )n-OH;
    R 13选自C 1-4的烷基取代或者未取代的苯基； R 13 is selected from C 1-4 alkyl substituted or unsubstituted phenyl;
    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 PCTCN2020133493-appb-100007
    Figure PCTCN2020133493-appb-100008
    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 PCTCN2020133493-appb-100007
    Figure PCTCN2020133493-appb-100008
    Figure PCTCN2020133493-appb-100009
    包括
    Figure PCTCN2020133493-appb-100010
    Figure PCTCN2020133493-appb-100009
    include
    Figure PCTCN2020133493-appb-100010
    其中，R 7和R 9如上定义。 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、环丙基、正丁基、异丁基、仲丁基、叔丁基、环丁基； The C 1-4 alkyl group is selected from methyl, ethyl, propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl;
    -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、环丙烷、正丁烷、异丁烷、仲丁烷、叔丁烷、环丁烷； The C 1-4 alkane is selected from methane, ethane, propane, isopropane, cyclopropane, n-butane, isobutane, sec-butane, tert-butane, cyclobutane;
    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 group, methanol, ethanol, n-propanol, 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选自氨基、甲胺、乙胺、丙胺、丁胺、二甲胺、甲乙胺、甲丙胺、甲丁胺、二乙胺、乙丙胺、乙丁胺、二丙胺、丙丁胺、二丁胺。 The NR 10 R 11 is selected from the group consisting of amino, methylamine, ethylamine, propylamine, butylamine, dimethylamine, methylethylamine, methylpropylamine, methylbutylamine, diethylamine, ethylenepropylamine, ethylbutylamine, dipropylamine, propylbutylamine Amine, dibutylamine.
  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 PCTCN2020133493-appb-100011
    Figure PCTCN2020133493-appb-100012
    More than one carbon atom of the C 3-12 fat is replaced by 0-2 N, O, S atoms, selected from:
    Figure PCTCN2020133493-appb-100011
    Figure PCTCN2020133493-appb-100012
  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 PCTCN2020133493-appb-100013
    Figure PCTCN2020133493-appb-100014
    R 2 is selected from benzene ring,
    Figure PCTCN2020133493-appb-100013
    Figure PCTCN2020133493-appb-100014
    Figure PCTCN2020133493-appb-100015
    Figure PCTCN2020133493-appb-100015
    Ar选自以下基团：
    Figure PCTCN2020133493-appb-100016
    Figure PCTCN2020133493-appb-100017
    Ar is selected from the following groups:
    Figure PCTCN2020133493-appb-100016
    Figure PCTCN2020133493-appb-100017
  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 PCTCN2020133493-appb-100018
    Figure PCTCN2020133493-appb-100018
    Figure PCTCN2020133493-appb-100019
    Figure PCTCN2020133493-appb-100019
    Figure PCTCN2020133493-appb-100020
    Figure PCTCN2020133493-appb-100020
    Figure PCTCN2020133493-appb-100021
    Figure PCTCN2020133493-appb-100021
    Figure PCTCN2020133493-appb-100022
    Figure PCTCN2020133493-appb-100022
    Figure PCTCN2020133493-appb-100023
    Figure PCTCN2020133493-appb-100023
    Figure PCTCN2020133493-appb-100024
    Figure PCTCN2020133493-appb-100024
    Figure PCTCN2020133493-appb-100025
    Figure PCTCN2020133493-appb-100025
  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 pharmaceuticals 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.
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