WO2021051827A1 - Apoptosis protein inhibitor, preparation method therefor, and use thereof - Google Patents

Abstract

The present invention relates to a compound as represented by formula I, and a racemate, a stereoisomer, a tautomer, an isotopic marker, nitrogen oxide, a solvate, a polymorph, a metabolite, an ester, a prodrug or a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising same, a preparation method therefor, and pharmaceutical use thereof. The structure of formula I is as follows.

Description

Inhibitor of cell apoptosis protein and preparation method and application thereof

This application claims the priority of the prior application filed with the State Intellectual Property Office of China on September 18, 2019. The patent application number is 201910883624.5 and the invention title is "A cell apoptosis protein inhibitor and its preparation method and use". The full text of this application is incorporated into this application by reference.

Technical field

The invention belongs to the field of medicine, and specifically relates to a cell apoptosis protein inhibitor and a preparation method and application thereof.

Background technique

Apoptosis refers to the orderly and autonomous death of cells controlled by genes in order to maintain the stability of the internal environment. It plays an important role in the evolution of organisms, the stability of the internal environment, and the development of multiple systems. Apoptosis signal transduction is divided into intrinsic (mediated by death receptor-ligand interaction) and extrinsic (mediated by cell stress and mitochondrial permeability). The two pathways finally converge in Caspase. Once the apoptotic signal is activated, caspase will cleave a large number of substrates related to cell death, causing cell death. Inhibitor of apoptosis (inhibitor of apoptosis proteins, IAPs) is a family of highly conserved endogenous anti-apoptotic factors, which inhibit cells mainly by inhibiting the activity of Caspase and participating in the mediation of nuclear factor NF-κB. Apoptosis. SMAC mimics (also known as IAP antagonists) are synthetic small molecules that mimic the structure of the four N-terminal amino acids of SMAC and IAP antagonist activity. When administered to animals suffering from proliferative diseases, the SMAC mimics antagonize IAP, leading to an increase in apoptosis in abnormally proliferating cells. There is a need to develop new IAP antagonists with better activity, selectivity and safety.

Summary of the invention

In order to solve the problems in the prior art, the present invention provides a compound represented by formula I and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, poly Crystal form, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof:

Wherein, R ₁ and R ₂ are independently selected from;

R ₃ , R ₄ and R ₅ are each independently selected from H or optionally substituted by 1, 2 or 3 R: (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one, two or more Heteroatomic (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, C _3-12 cycloalkyl group, 3-12 membered heterocyclic group, C _6-20 aryl group or 5-14 membered heteroaryl group;

Ring A and Ring B are each independently selected from: C _6-20 aryl or 5-14 membered heteroaryl;

R ₆ and R ₇ are each independently selected from halogen, hydroxyl, or selected from optionally substituted by 1, 2 or 3 R: (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one, two or more A heteroatom (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, a C _3-12 cycloalkyl group, a 3-12 membered heterocyclic group, a C _6-20 aryl group or a 5-14 membered heteroaryl group;

m and n are independently selected from: 0, 1, 2 or 3;

R is selected from halogen, CN, OH, SH, NH ₂ , COOH, or selected from optionally substituted by 1, 2 or 3 R': (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one or two _{(C 1} -C ₁₂ ) aliphatic hydrocarbon group with one or more heteroatoms _{, C 3-12} cycloalkyl, 3-12 membered heterocyclic group, C _6-20 aryl group or 5-14 membered heteroaryl group;

R'is selected from halogen, CN, OH, SH, NH ₂ , COOH; P ₁ and P ₂ are independently selected from halogen, OH, CN, NH ₂ , COOH, (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally _{A (C 1} -C ₁₂ ) aliphatic hydrocarbon group containing one, two or more heteroatoms.

W is selected from a single bond, -O-, -S-, -NH- or optionally substituted by 1, 2 or 3 R: (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one, two or (C ₁ -C ₁₂ ) aliphatic hydrocarbon groups with _{more heteroatoms, C 3-12} cycloalkyl groups, 3-12 membered heterocyclic groups, C _6-20 aryl groups or 5-14 membered heteroaryl groups.

According to the embodiment of the present invention,

_{In the "(C 1} -C ₁₂ )aliphatic hydrocarbon group optionally containing one, two or more heteroatoms", the heteroatom may be selected from sulfur, nitrogen, oxygen, phosphorus and silicon, optionally , The heteroatom is inserted into the aliphatic hydrocarbon group, optionally CC bond and CH bond. ; For example, it can be selected from (C ₁ -C ₁₂ ) aliphatic hydrocarbyloxy group, (C ₁ -C ₁₂ ) aliphatic hydrocarbyl mercapto group, (C ₁ -C ₆ ) aliphatic hydrocarbyloxy group, (C ₁ -C ₆ ) aliphatic hydrocarbyl mercapto group , (C ₁ -C ₆ ) aliphatic hydrocarbyloxy group (C ₁ -C ₆ ) aliphatic hydrocarbyl group, (C ₁ -C ₆ ) aliphatic hydrocarbyl mercapto group (C ₁ -C ₆ ) aliphatic hydrocarbyl group, N-(C ₁ -C ₃₎ ) Aliphatic hydrocarbon group (C ₁ -C ₆ ) aliphatic hydrocarbon group, N,N-bis-(C ₁ -C ₃ ) aliphatic hydrocarbon group (C ₁ -C ₆ ) aliphatic hydrocarbon group;

The (C ₁ -C ₁₂ ) aliphatic hydrocarbon group may be selected from (C ₁ -C ₁₂ )alkyl, (C ₂ -C ₁₂ )alkenyl, (C ₂ -C ₁₂ )alkynyl, and in some embodiments, Can be selected from (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl;

According to the embodiments of the present invention, in some embodiments, the R ₃ , R ₄ and R ₅ may be independently selected from the following groups: methyl, ethyl, n-propyl, isopropyl, n-butyl , Isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 1-ethylvinyl, 1-methyl-2-propenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 2-methyl-2-propenyl, 1 -Pentenyl, 1-hexenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 1-methyl-2-propynyl, 3-butynyl, 1 -Pentynyl, 1-hexynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,

The "halogen" is selected from F, Cl, Br, I;

According to an embodiment of the present invention, the structure of formula I is further selected from the following formula II, formula III, formula IV, and formula V:

In the formula II, formula III, formula IV, and formula V, R ₁ , R ₂ , R ₆ , R ₇ , P ₁ , P ₂ , W, n, m, ring A, and ring B are as defined in the above formula I .

According to the present invention, in the above formula IV, R ₁ and R ₂ are independently selected from;

R ₃ , R ₄ and R ₅ are each independently selected from H or the following groups optionally substituted by 1, 2 or 3 R: (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one, two or (C ₁ -C ₁₂ ) aliphatic hydrocarbon groups with _{more heteroatoms, C 3-12} cycloalkyl groups, 3-12 membered heterocyclic groups;

Ring A and Ring B are each independently selected from: C _6-20 aryl or 5-14 membered heteroaryl;

R ₆ and R ₇ are each independently selected from halogen or hydroxyl;

m and n are independently selected from: 0, 1, 2 or 3;

R is selected from halogen, CN, OH, SH, NH ₂ , COOH, or selected from optionally substituted by 1, 2 or 3 R': (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one or two _{(C 1} -C ₁₂ ) aliphatic hydrocarbon group with one or more heteroatoms _{, C 3-12} cycloalkyl, 3-12 membered heterocyclic group, C _6-20 aryl group or 5-14 membered heteroaryl group;

R'is selected from halogen, CN, OH, SH, NH ₂ , COOH;

P ₁ and P ₂ are each independently selected from halogen, OH, NH ₂ , and (C ₁ -C ₁₂ ) aliphatic hydrocarbon groups.

W is selected from a single bond, -O-, -S-, -NH-.

Illustrative, non-limiting specific examples of the compounds of the present invention are as follows:

The present invention also provides the compound represented by formula I (including the compound of formula II-form V) and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, poly The preparation methods of crystalline forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof are not limited to the methods described below. All raw materials are prepared according to the group characteristics of the target molecule in accordance with the general formula, and are prepared through the schemes in these routes and methods well known to those of ordinary skill in the organic chemistry field or directly purchased. The compounds of the present invention can be synthesized by combining the following methods with synthetic methods known in the field of synthetic organic chemistry or related modification methods recognized by those skilled in the art.

In the first scheme, the preparation of the compound of the present invention includes the following steps:

In the second scheme, the preparation of the compound of the present invention includes the following steps:

In the scheme, M-9 can be obtained by using the preparation steps in the first scheme (M-9 is prepared from M-1 as a starting material).

According to the embodiments of the present invention, of the two solutions,

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , P ₁ , P ₂ , and W are as defined in formula I;

R ₃ ′ is selected from groups within the range defined by _{R 3} , and independently selected from groups different _{from R 3;}

PG ₁ and PG ₂ represent different amino protecting groups, which can be selected from the group consisting of tert-butoxycarbonyl (N-boc), benzyloxycarbonyl protecting group (N-cbz), fluorene methoxycarbonyl protecting group (N-Fmoc ), preferably, PG _{1 is} selected from tert-butoxycarbonyl, and PG _{2 is} selected from benzyloxycarbonyl protecting groups.

According to an embodiment of the present invention, in the two preparation schemes, M-1 can be used as a starting material to prepare M-9, and the preparation of M-9 includes the following steps:

(a1) Taking N-PG ₁ protected prolinol M-1 as the starting material, it reacts with phthalimide under the action of triphenylphosphine and diisopropyl azodicarboxylate to form M- 2;

Preferably, in the step, the reaction temperature can be selected from 0°C to 25°C, and the reaction solvent can be selected from inert aprotic solvents, for example, selected from tetrahydrofuran, toluene, dichloromethane, etc.;

(a2) M-2 hydrazinolysis under hydrazine hydrate conditions to obtain M-3;

Preferably, in the step, the reaction temperature can be selected from 25°C to 80°C, and the reaction solvent can be selected from methanol, ethanol, tetrahydrofuran, etc.;

(a3) M-3 reacts with M-4 under alkaline conditions to produce M-5;

Preferably, in the step, the reaction temperature can be selected from 70°C to 90°C, the reaction solvent can be selected from acetonitrile, DMF, tetrahydrofuran, etc., and the alkaline conditions can be selected from alkali metal or alkaline earth metal carbonates, such as Potassium carbonate, sodium carbonate, etc.;

(a4) M-5 is reduced by iron powder to obtain M-6;

Preferably, in the step, the reaction temperature can be selected from room temperature, for example, 25°C, and the reaction solvent can be selected from acetic acid;

(a5) M-6 and M-7 are ring closed to obtain M-8 in an acidic system;

Preferably, in the step, hydrochloric acid can be added to the acidic system, for example, a 4N hydrochloric acid aqueous solution can be added;

(a6) The hydroxyl group of M-8 is oxidized to obtain the aldehyde compound M-9;

Preferably, in the step, the oxidizing agent used is Des Martin reagent, the reaction temperature can be selected from 0°C to 25°C, and the reaction solvent can be selected from inert aprotic solvents, such as tetrahydrofuran, dichloromethane, toluene, etc.;

According to an embodiment of the present invention, in the first solution, the process of preparing Formula I from M-9 includes the following steps:

(b1) Rings of M-9 and M-10 are closed under catalyst conditions to obtain M-11;

Preferably, in the step, the catalyst can be selected from Oxone, sodium metabisulfite, etc., the reaction temperature can be selected from 25°C to 140°C, and the reaction solvent can be selected from inert high boiling point solvents, such as DMSO, DMF, etc.;

(b2) M-11 removes the protecting group to obtain M-12;

Preferably, in the step, the reaction solvent can be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the conditions for removing the protective group can be selected from acidic conditions. For example, when the protective group is selected from N-boc, you can select In the presence of palladium on carbon, for example, when the protecting group is selected from N-cbz, it can be selected from the presence of piperidine, for example, when the protecting group is selected from N-fmoc;

(b3) M-12 undergoes condensation reaction with M-13 in an inert organic solvent under the action of a condensing agent to obtain M-14;

Preferably, in the step, the inert organic solvent can be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature can be selected from 25°C to 120°C, and the condensing agent can be selected from N, N '-Diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2-(7-azobenzotriazole)- N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate, etc.;

(b4) M-14 removes the protecting group to obtain M-15;

Preferably, in the step, the conditions for removing the protecting group can be selected from acidic conditions, for example, when the protecting group is selected from N-boc, it can be selected from the presence of palladium on carbon, for example, when the protecting group is selected from N-cbz, it can be selected. In the presence of piperidine, for example, when the protecting group is selected from N-fmoc; the reaction solvent is selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc.;

(b5) M-15 undergoes condensation reaction with M-16 in an inert organic solvent under the action of a condensing agent to obtain M-17;

Preferably, in the step, the inert organic solvent can be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature can be selected from 25°C to 120°C, and the condensing agent can be selected from N, N '-Diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2-(7-azobenzotriazole)- N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate, etc.;

(b6) M-17 removes the protective group to obtain the target compound of formula I;

Preferably, the reaction solvent in the reaction is tetrahydrofuran, dichloromethane, methanol, toluene and the like.

According to an embodiment of the present invention, in the second embodiment, the process of preparing Formula I from M-9 includes the following steps:

(c1) M-9 and N-1 are ring closed under catalyst conditions to obtain N-2;

Preferably, in the step, the catalyst can be selected from Oxone, sodium metabisulfite, etc., the reaction temperature can be selected from 25°C to 140°C, and the reaction solvent can be selected from inert high boiling point solvents, such as DMSO, DMF, etc.;

(c2) N-2 selectively removes a protecting group to obtain N-3;

Preferably, in the step, the reaction solvent can be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the conditions for removing the protective group can be selected from acidic conditions. For example, when the protective group is selected from N-boc, you can select In the presence of palladium on carbon, for example, when the protecting group is selected from N-cbz, it can be selected from the presence of piperidine, for example, when the protecting group is selected from N-fmoc;

(c3) N-3 undergoes condensation reaction with N-4 in an inert organic solvent under the action of a condensing agent to obtain N-5;

Preferably, in the step, the inert organic solvent can be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature can be selected from 25°C to 120°C, and the condensing agent can be selected from N, N' -Diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2-(7-azobenzotriazole)-N ,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate, etc.;

(c4) N-5 selectively removes a protecting group to obtain N-6;

Preferably, in the step, the reaction solvent can be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the conditions for removing the protective group can be selected from acidic conditions. For example, when the protective group is selected from N-boc, you can select In the presence of palladium on carbon, for example, when the protecting group is selected from N-cbz, it can be selected from the presence of piperidine, for example, when the protecting group is selected from N-fmoc;

(c5) N-6 undergoes condensation reaction with N-7 in an inert organic solvent under the action of a condensing agent to obtain N-8;

Preferably, in the step, the inert organic solvent can be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature can be selected from 25°C to 120°C, and the condensing agent can be selected from N, N '-Diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2-(7-azobenzotriazole)- N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate, etc.;

(c6) N-8 removes the protective group to obtain N-9;

Preferably, in the step, the reaction solvent can be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the conditions for removing the protective group can be selected from acidic conditions. For example, when the protective group is selected from N-boc, you can select In the presence of palladium on carbon, for example, when the protecting group is selected from N-cbz, it can be selected from the presence of piperidine, for example, when the protecting group is selected from N-fmoc;

(c7) N-9 undergoes condensation reaction with chiral amino acid compounds protected by _{PG 2} in an inert organic solvent under the action of a condensing agent to obtain compound N-10;

Preferably, in the step, the inert organic solvent can be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature can be selected from 25°C to 120°C, and the condensing agent can be selected from N, N '-Diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2-(7-azobenzotriazole)- N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate, etc.;

(c8) N-10 removes the protective group to obtain the target compound of formula I;

Preferably, in the step, the reaction solvent can be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the conditions for removing the protective group can be selected from acidic conditions. For example, when the protective group is selected from N-boc, you can select In the presence of palladium on carbon, for example, when the protecting group is selected from N-cbz, it can be selected from the presence of piperidine, for example, when the protecting group is selected from N-fmoc.

The present invention further provides a pharmaceutical composition comprising the compound of formula I of the present invention and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, and polymorphs. Forms, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof.

In some embodiments, the pharmaceutical composition of the present invention further comprises a therapeutically effective amount of the compound of formula I of the present invention and its racemates, stereoisomers, tautomers, isotope markers, nitroxides Substances, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof and pharmaceutically acceptable carriers.

The carrier in the pharmaceutical composition is "acceptable", which is compatible with the active ingredient of the composition (and preferably capable of stabilizing the active ingredient) and is not harmful to the subject being treated. One or more solubilizers can be used as pharmaceutical excipients for the delivery of active compounds.

Therefore, the present invention further provides the compound of formula I and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, metabolites, esters, pro The use of the drug or its pharmaceutically acceptable salt in the preparation of an apoptosis protein inhibitor.

The present invention further provides the compound of formula I and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or Use of its pharmaceutically acceptable salt in medicine for treating diseases or disorders caused by IAP disorders.

As SMAC mimics, the compounds of the present invention can be used to treat proliferative diseases, such as a variety of benign tumors or malignant tumors (cancer), benign proliferative diseases (such as psoriasis, benign prostatic hypertrophy and restenosis), or autoimmune diseases ( For example, autoimmune proliferative glomerulonephritis, lymphoid tissue proliferative autoimmune response). Cancers that can be treated with IAP antagonists include but are not limited to one or more of the following cancers: lung adenocarcinoma, pancreatic cancer, colon cancer, ovarian cancer, breast cancer, mesothelioma, peripheral neuroma, bladder cancer , Glioblastoma, melanoma, adrenal cortical carcinoma, AIDS-related lymphoma, anal cancer, bladder cancer, meningioma, glioma, astrocytoma, breast cancer, cervical cancer, Chronic myelodysplastic disorders (e.g., chronic lymphocytic leukemia, chronic myelogenous leukemia), colon cancer, endocrine adenocarcinoma, endometrial cancer, ependymoma, esophageal cancer, Ewing’s sarcoma, extracranial germ cell tumor, Extragonadal germ cell tumor, extrahepatic cholangiocarcinoma, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gestational trophoblastoma, hairy cell leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hypopharyngeal carcinoma, Intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, laryngeal cancer, leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, lip cancer, oral cancer, liver cancer, male breast cancer, malignant mesothelioma , Medulloblastoma, melanoma, Merkel cell carcinoma, metastatic squamous neck cancer, multiple myeloma and other plasma cell neoplasms, mycosis fungoides and Sezary syndrome (Sezary syndrome), myelodysplastic syndrome, nasopharyngeal carcinoma, neuroblastoma, non-small cell lung cancer, small cell lung cancer, oropharyngeal cancer, bone cancer (including osteosarcoma and bone malignant fibrous histiocytoma), ovarian epithelial cancer , Ovarian germ cell tumor, ovarian low malignant potential tumors, pancreatic cancer, paranasal sinus cancer, parathyroid cancer, penile cancer, pheochromocytoma, pituitary tumor, prostate cancer, rectal cancer , Renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, small intestine cancer, soft tissue sarcoma, supratentorial primary neuroectodermal tumor, pineoblastoma, testicular cancer, thymoma, thymic cancer , Thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer, as well as Wilm's tumor and other childhood kidney tumors.

Therefore, the present invention further provides the compound of formula I and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, metabolites, esters, pro The use of the drug or a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of a medicament for preventing and/or treating tumors or tumor-related disorders, the tumors or tumor-related disorders are as described above.

The method of the present invention may include administering the compound of the present invention alone, as well as administering the compound of the present invention in combination with one or more other chemotherapeutic agents. The administration of multiple drugs can be performed simultaneously or sequentially.

Term explanation:

Unless otherwise stated, the definitions of groups and terms described in the specification and claims of this application include definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples. And so on, can be combined and combined with each other arbitrarily. Such combination and the group definition and compound structure after the combination should fall within the scope of the specification of this application.

The numerical range described in the specification and claims of the present application, when the numerical range is defined as an "integer", should be understood as recording the two end points of the range and each integer in the range. For example, "an integer from 0 to 6" should be understood as recording each integer of 0, 1, 2, 3, 4, 5, and 6. "More" means three or more.

The term "halogen" refers to F, Cl, Br, and I. In other words, F, Cl, Br, and I can be described as "halogen" in this specification.

The term "aliphatic hydrocarbon group" includes saturated or unsaturated, linear or branched chain or cyclic hydrocarbon groups. The type of the aliphatic hydrocarbon group can be selected from alkyl, alkenyl, alkynyl, etc. The carbon atoms of the aliphatic hydrocarbon group The number is preferably 1-12, and can also be 1-10, and a further preferred range is 1-6, which can specifically include but not limited to the following groups: methyl, ethyl, n-propyl, isopropyl, n-butyl , Isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 1-ethylvinyl, 1-methyl-2-propenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 2-methyl-2-propenyl, 1 -Pentenyl, 1-hexenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 1-methyl-2-propynyl, 3-butynyl, 1 -Pentynyl, 1-hexynyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl;

The "aliphatic hydrocarbon group" may optionally include one, two or more heteroatoms (or construed as optional heteroatoms inserted into the aliphatic hydrocarbon group, optionally CC bonds and CH bonds). Suitable heteroatoms are obvious to those skilled in the art and include, for example, sulfur, nitrogen, oxygen, phosphorus, and silicon. The heteroatom-containing aliphatic hydrocarbon group can be selected from the following groups: (C ₁ -C ₆ ) aliphatic hydrocarbon group oxy group, (C ₁ -C ₆ ) aliphatic hydrocarbon group mercapto group, (C ₁ -C ₆ ) aliphatic hydrocarbon group oxygen Group (C ₁ -C ₆ ) aliphatic hydrocarbon group, (C ₁ -C ₆ ) aliphatic hydrocarbon group mercapto group (C ₁ -C ₆ ) aliphatic hydrocarbon group, N-(C ₁ -C ₃ ) aliphatic hydrocarbon group amine group (C ₁ -C ₆₎ ) Aliphatic hydrocarbon group, N,N-bis-(C ₁ -C ₃ ) aliphatic hydrocarbon amino (C ₁ -C ₆ ) aliphatic hydrocarbon group, such as methoxy, ethoxy, propoxy, butoxy, Pentyloxy, methoxymethyl, ethoxymethyl, propoxymethyl, methoxyethyl, ethoxyethyl, propoxyethyl, methoxypropyl, ethoxypropyl Group, propoxypropyl, N-methylaminomethyl, N-methylaminoethyl, N-ethylaminoethyl, N,N-dimethylaminomethyl, N,N-dimethyl Aminoethyl, N,N-diethylaminoethyl; the "aliphatic hydrocarbon group" contained in other groups is the same as explained above.

The term "C _3-12 cycloalkyl" should be understood to mean a saturated or unsaturated monovalent monocyclic or bicyclic ring, which has 3-12 carbon atoms, preferably "C _3-10 cycloalkyl". The term "C _3-10 cycloalkyl" should be understood to mean a saturated or unsaturated monovalent monocyclic or bicyclic ring having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. The C _3-10 cycloalkyl group may be a monocyclic hydrocarbon group, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic group. Hydrocarbon groups such as tetralin or decalin.

The term "3-12 membered heterocyclic group" means a saturated or unsaturated monovalent monocyclic or bicyclic ring, which contains 1-5 heteroatoms independently selected from N, O and S, and heteroatom-containing groups do not have Aromatic, the 3-12 membered heterocyclic group is preferably "3-10 membered heterocyclic group". The term "3-10 membered heterocyclic group" means a saturated monovalent monocyclic or bicyclic ring, which contains 1-5, preferably 1-3 heteroatoms selected from N, O and S. The heterocyclic group may be connected to the rest of the molecule through any of the carbon atoms or the nitrogen atom (if present). In particular, the heterocyclic group may include but is not limited to: 4-membered ring, such as azetidinyl, oxetanyl; 5-membered ring, such as tetrahydrofuranyl, tetrahydrothienyl, dioxane Pentenyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholine Group, piperazinyl, or trithiaalkyl; or 7-membered ring, such as diazacycloheptanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclic group may be bicyclic, such as but not limited to a 5, 5-membered ring, such as hexahydrocyclopenta[c]pyrrole-2(1H)-yl ring, or a 5, 6-membered bicyclic ring, such as hexahydropyrrole And [1,2-a]pyrazine-2(1H)-yl ring. The ring containing the nitrogen atom may be partially unsaturated, that is, it may contain one or more double bonds, such as but not limited to 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadi Azinyl, 4,5-dihydrooxazolyl or 4H-[1,4]thiazinyl, or it may be benzo-fused, such as but not limited to dihydroisoquinolinyl. According to the present invention, the 3-12 membered heterocyclic group may be further selected from the following groups:

The term "C _6-20 aryl" should be understood to preferably mean a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring with 6-20 carbon atoms, preferably "C _6-14 aryl" . The term "C _6-14 aryl" should be understood as preferably meaning a monocyclic, bicyclic or partially aromatic monocyclic or partially aromatic monocyclic or partially aromatic having 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms A tricyclic hydrocarbon ring ("C _6-14 aryl"), especially a ring having 6 carbon atoms ("C ₆ aryl"), such as phenyl; or biphenyl, or one having 9 carbon atoms Ring ("C ₉ aryl"), such as indanyl or indenyl, or a ring with 10 carbon atoms ("C ₁₀ aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, Either a ring having 13 carbon atoms ("C ₁₃ aryl"), such as fluorenyl, or a ring having 14 carbon atoms ("C ₁₄ aryl"), such as anthracenyl.

The term "5-14 membered heteroaryl" should be understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, especially 5 or 6 or 9 or 10 carbon atoms, and it contains 1 to 5, preferably 1 to 3 heteroatoms independently selected from N, O and S, and, in addition, in each In the case it can be benzo-fused. In particular, the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiol Diazolyl, thio-4H-pyrazolyl, etc. and their benzo derivatives, such as benzofuranyl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzene O-triazolyl, indazolyl, indolyl, isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and their benzo derivatives, such as quinoline Group, quinazolinyl, isoquinolinyl, etc.; or azecinyl, indazinyl, purinyl, etc. and their benzo derivatives; or cinolinyl, phthalazinyl, quinazolinyl, quinoxa Linyl, naphthyridinyl, pterridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, etc.

Unless otherwise specified, heterocyclic groups or heteroaryl groups include all possible isomeric forms thereof, such as positional isomers thereof. Therefore, for some illustrative non-limiting examples, pyridinyl or pyridinylene includes pyridin-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-3-yl, pyridin-4-yl And pyridin-4-yl; thienyl or thienylene includes thiophen-2-yl, thiophen-2-yl, thiophen-3-yl, and thiophen-3-yl.

According to its molecular structure, the compound of the present invention may be chiral, and therefore may exist in various enantiomeric forms. Therefore, these compounds may exist in racemate form or optically active form. The compounds of the present invention or intermediates thereof can be separated into enantiomeric compounds by chemical or physical methods known to those skilled in the art, or used in synthesis in this form. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with optically active resolving reagents. Examples of suitable resolution reagents are optically active acids such as R and S forms of tartaric acid, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid, appropriate N-protected amino acids (e.g., N- Benzoyl proline or N-benzenesulfonyl proline) or various optically active camphor sulfonic acids. With the aid of optically active resolving reagents (such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chiral derivatized methacrylate polymers) fixed on silica gel, Chromatographic enantiomeric resolution is advantageously performed. Suitable eluents for this purpose are aqueous or alcohol-containing solvent mixtures, for example, hexane/isopropanol/acetonitrile.

Those skilled in the art will understand that since nitrogen needs to have available lone pairs of electrons for being oxidized to nitrogen oxides, not all nitrogen-containing heterocycles can form N-oxides; those skilled in the art will recognize that N-oxides can be formed. -Nitrogen-containing heterocycles of oxides. Those skilled in the art will also recognize that tertiary amines can form N-oxides. The synthetic methods for preparing heterocyclic and tertiary amine N-oxides are well known to those skilled in the art, and the synthetic methods include the use of peroxyacids such as peroxyacetic acid and m-chloroperoxybenzoic acid (MCPBA), peroxy Hydrogen oxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate and dioxirane (dioxirane) such as dimethyldioxirane oxidize heterocycles and tertiary amines. These methods of preparing N-oxides have been extensively described and reviewed in the literature.

The pharmaceutically acceptable salt may be, for example, an acid addition salt of the compound of the present invention that has a nitrogen atom in the chain or ring and is sufficiently basic, for example, an acid addition salt formed with the following inorganic acids: for example, hydrochloric acid, hydrofluorine Acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid or nitric acid, or hydrogen sulfate, or acid addition salts formed with the following organic acids: for example, formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid , Propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphor acid, cinnamic acid, cyclopentane Propionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, niacin, hexanoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonate Acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalene disulfonic acid, Camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid , Ascorbic acid, glucoheptanoic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid or thiocyanic acid.

In addition, another suitable pharmaceutically acceptable salt of the compound of the present invention that is sufficiently acidic is an alkali metal salt (such as a sodium salt or potassium salt), an alkaline earth metal salt (such as a calcium salt or a magnesium salt), an ammonium salt, Or salts formed with organic bases that provide physiologically acceptable cations, such as salts formed with sodium ions, potassium ions, N-methylglucamine, dimethylglucamine, ethylglucosamine, Lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serinol, trihydroxymethylaminomethane, aminopropanediol, 1-amino-2 ,3,4-Butanetriol. As an example, the pharmaceutically acceptable salt includes the salt formed by the group -COOH with the following substances: sodium ion, potassium ion, calcium ion, magnesium ion, N-methylglucamine, dimethylglucamine, Ethyl glucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serinol, trishydroxymethylaminomethane, aminopropanediol , 1-Amino-2,3,4-butanetriol.

In addition, basic nitrogen-containing groups can be quaternized with the following reagents: lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates, such as sulfuric acid Dimethyl, diethyl sulfate, dibutyl sulfate and dipentyl sulfate; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl Halides such as benzyl and phenethyl bromide. As examples, pharmaceutically acceptable salts include hydrochloride, sulfate, nitrate, bisulfate, hydrobromide, acetate, oxalate, citrate, methanesulfonate, formate, or Meglumine salt and so on.

Since the compound of the present invention may have multiple salt-forming sites, the "pharmaceutically acceptable salt" includes not only the salt formed at one salt-forming site of the compound of the present invention, but also 2, 3 or all of them. The salt formed at the salt-forming site. For this reason, the molar ratio of the compound of formula (I) to the acid root ion (anion) or base cation required for salt formation in the "pharmaceutically acceptable salt" can be varied within a relatively large range, for example, it can be 4 :1～1:4, such as 3:1, 2:1, 1:1, 1:2, 1:3, etc.

According to the present invention, pharmaceutically acceptable anions include anions selected from the ionization of inorganic acids or organic acids. The "inorganic acid" includes, but is not limited to, hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid, or nitric acid. The "organic acid" includes but is not limited to formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid , 2-(4-Hydroxybenzoyl)benzoic acid, camphor acid, cinnamic acid, cyclopentane propionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, hexanoic acid, pectinic acid , Persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecyl sulfuric acid, ethanesulfonic acid, benzene Sulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, Adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptanoic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid, or thiocyanic acid .

According to the positions and properties of different substituents, the compounds of the present invention may also contain one or more asymmetric centers. Asymmetric carbon atoms can exist in the (R) or (S) configuration. When there is only one asymmetric center, a racemic mixture is produced, and when multiple asymmetric centers are contained, a diastereomeric mixture is obtained. In some cases, there may be asymmetry due to hindered rotation around a specific bond, for example, the central bond connects two substituted aromatic rings of a specific compound. In addition, the substituent may also exist in a cis- or trans-isomeric form.

The compounds of the present invention also include all possible stereoisomers of each, which is a single stereoisomer or the stereoisomer (for example, R-isomer or S-isomer, or E-isomer or Z-isomer) in the form of any mixture in any ratio. A single stereoisomer (e.g., single enantiomer or single diastereomer) of the compound of the present invention can be achieved by any suitable prior art method (e.g., chromatography, especially, e.g., chiral chromatography) Separation.

The term "tautomer" refers to an isomer of a functional group resulting from the rapid movement of an atom in a molecule at two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist in two or more mutually convertible species. Proton shift tautomers result from the migration of covalently bonded hydrogen atoms between two atoms. Tautomers generally exist in an equilibrium form. An attempt to separate a single tautomer usually produces a mixture whose physical and chemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties of the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the ketone type is dominant; in phenol, the enol type is dominant. The present invention encompasses all tautomeric forms of the compound.

In the present invention, the involved compounds also include isotopically-labeled compounds. The isotopically-labeled compounds are the same as those shown in Formula I, but one or more of the atoms are different from the usual atomic mass or mass number. Naturally occurring atomic mass or mass number atomic substitution. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of H, C, N, O, S, F, and Cl, such as ² H, ³ H, ¹³ C, ¹¹ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, respectively. , ¹⁷ O, ³² P, ³⁵ S, ¹⁸ F and ³⁶ Cl. The compounds of the present invention containing the above-mentioned isotopes and/or other isotopes of other atoms, prodrugs thereof, or pharmaceutically acceptable salts of the compounds or prodrugs are within the scope of the present invention. Certain isotopically-labeled compounds of the present invention, such as compounds incorporating radioisotopes (such as ³ H and ¹⁴ C), can be used for drug and/or substrate tissue distribution determination. Tritium (ie ³ H) and carbon 14 (ie ¹⁴ C) isotopes are particularly preferred due to their ease of preparation and detectability. Furthermore, the replacement of heavier isotopes (such as deuterium, ie ² H) can provide certain therapeutic advantages derived from higher metabolic stability (for example, increased in vivo half-life or reduced dosage requirements), and therefore can be used in certain Some cases are preferred. The compounds of the present invention as claimed in the claims can be specifically limited to be substituted with deuterium or tritium. In addition, the absence of the term deuterium or tritium in the hydrogen appearing in the substituent does not mean that deuterium or tritium is excluded, but deuterium or tritium may also be included in the same way.

The term "effective amount" or "therapeutically effective amount" refers to the amount of the compound of the present invention sufficient to achieve the intended application (including but not limited to the treatment of diseases as defined below). The therapeutically effective amount may vary depending on the following factors: the intended application (in vitro or in vivo), or the subject to be treated and the disease condition such as the weight and age of the subject, the severity of the disease condition and the mode of administration, etc. It can be easily determined by a person of ordinary skill in the art. The specific dosage will vary depending on the following factors: the particular compound selected, the dosing regimen on which it is based, whether it is administered in combination with other compounds, the timing of administration, the tissue to be administered, and the physical delivery system carried.

The term "solvate" refers to those forms of the compound of the present invention, which form a complex by coordination with solvent molecules in a solid or liquid state. Hydrates are a specific form of solvates in which the coordination is carried out with water. In the present invention, the preferred solvate is a hydrate. Further, the pharmaceutically acceptable solvate (hydrate) of the compound of the general formula I of the present invention refers to the co-crystal and clathrate formed by the compound I and one or more stoichiometric molecules of water or other solvents. Solvents that can be used for solvates include, but are not limited to: water, methanol, ethanol, ethylene glycol, and acetic acid.

The term "prodrug" or "prodrug" represents the conversion of a compound into a compound represented by the aforementioned general formula or specific compound in vivo. Such conversion is affected by the hydrolysis of the prodrug in the blood or the enzymatic conversion of the prodrug into the maternal structure in the blood or tissues. The prodrugs of the present invention can be esters. In the present invention, esters can be used as prodrugs including phenyl esters, aliphatic (C1-24) esters, acyloxymethyl esters, carbonates, carbamates and amino acids. Esters. For example, a compound in the present invention contains a hydroxyl/carboxyl group, that is, it can be acylated to obtain a compound in the form of a prodrug. Other prodrug forms include phosphate esters, for example, these phosphate ester compounds are obtained by phosphorylation of the parent hydroxyl group.

Beneficial effect

1) The compound of the present invention adopts a novel structure of general formula I, has good binding affinity to XIAP, cIAP1, and cIAP2 protein, has good IAP inhibitory activity, and the inhibitory effect of some compounds is significantly better than the positive control drug;

2) The compound of the present invention has a good inhibitory effect on cell growth in MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines, and the inhibitory effect of some compounds is significantly better than the positive control drug.

detailed description

The technical solution of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only illustrative and explanation of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the foregoing contents of the present invention are covered by the scope of the present invention.

Unless otherwise specified, the raw materials and reagents used in the following examples are all commercially available products, or can be prepared by known methods. In the following specific examples, the acquisition process involving chiral compounds all include the steps of preparation, separation and purification using a chiral column.

Example 1: Preparation of benzyloxycarbonyl (2S)-2-[(1,3-dioxoisoindol-2-yl)methyl]pyrrolidine-1-carboxylate

Take 93.5g (0.398mol) of N-Cbz-L-prolinol, 70.2g (0.478mol) of phthalimide and 104.3g (0.398mol) of triphenylphosphine in a 2L three-necked reaction flask, add tetrahydrofuran (1.3L), under the protection of nitrogen, the system was cooled to 0°C, and 80.4g (0.398mol) of DIAD was added dropwise at 0°C. Then the system was heated to room temperature and stirred for 16 hours. TLC detected that the reaction was complete. 400 mL of water was added for quenching, stirring was continued for 30 minutes, and extracted with ethyl acetate (300 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:40) to obtain benzyloxycarbonyl (2S)-2-[(1,3-dioxoisoindol-2-yl) Methyl]pyrrolidine-1-carboxylate (98.5 g, 68%).

Example 2: Preparation of benzyloxycarbonyl (2S)-2-(aminomethyl)pyrrolidine-1-carboxylate

Take 65.5g (0.18mol) of benzyloxycarbonyl(2S)-2-[(1,3-dioxoisoindol-2-yl)methyl]pyrrolidine-1-carboxylate in a 1L reaction flask, Add 450 mL of ethanol, heat to 80°C, and add 17 g (0.45 mol, 85%) of hydrazine hydrate dropwise. Then it was stirred at 80°C for 12 hours. TLC detected that the reaction was complete. It was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain benzyloxycarbonyl (2S)-2-(aminomethyl)pyrrolidine-1-carboxylate (38.7 g, 92%), which was directly used in the next reaction.

Example 3: Preparation of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-nitro-anilino)methyl]pyrrolidine-1-carboxylate

Take 35 g (0.15 mol) of benzyloxycarbonyl (2S)-2-(aminomethyl) pyrrolidine-1-carboxylate in a 500 mL reaction flask, add 350 mL of acetonitrile, and add 41.4 g (0.3 mol) of potassium carbonate to it , Take 26.2g (0.17mol) of 1,4-difluoro-2-nitrobenzene and add it. Under the protection of nitrogen, the temperature was raised to 80°C and stirred for 1 hour. TLC detected that the reaction was complete, quenched by adding 200 mL of water, and extracted with ethyl acetate (200 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:40) to obtain benzyloxycarbonyl (2S)-2-[(4-fluoro-2-nitro-anilino)methyl] Pyrrolidine-1-carboxylate (49.2 g, 88%).

Example 4: Preparation of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]pyrrolidine-1-carboxylate

Take 49.2g (0.13mol) of benzyloxycarbonyl(2S)-2-[(4-fluoro-2-nitro-anilino)methyl]pyrrolidine-1-carboxylate into a 1L reaction flask, add 400mL acetic acid 135g of iron powder was added to it, stirred at room temperature for 16 hours, TLC detected that the reaction was complete, filtered, the filter cake was rinsed with ethyl acetate, the filtrate was concentrated under reduced pressure, 300mL of water was added, and ethyl acetate (200mL×3) was added to extract. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]pyrrolidine-1-carboxylate (43.5g, 96%) was obtained, which was directly used in the next reaction.

Example 5: (S)-Benzyl 2-((5-fluoro-2-(hydroxymethyl)-1H-benzo[D]imidazol-1-yl)methyl)pyrrolidine-1-carboxylate Preparation

Take 30g (87.4mmol) of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]pyrrolidine-1-carboxylate in a 500mL reaction flask, add 300mL of 4N hydrochloric acid, and Add 20g (0.262mol) of acetic alcohol into it, heat to 100°C and stir for 6 hours. TLC detects that the reaction is complete. Cool down to 0°C. Add saturated sodium bicarbonate solution to adjust pH=8-9, filter, and filter cake with methyl Wash with tert-butyl ether and dry to obtain (S)-benzyl 2-((5-fluoro-2-(hydroxymethyl)-1H-benzo[D]imidazol-1-yl)methyl)pyrrolidine-1 -Carboxylic acid ester (29g, 87%), used directly in the next reaction.

Example 6: Preparation of (S)-benzyl 2-((5-fluoro-2-formyl-1H-benzo[D]imidazol-1-yl)methyl)pyrrolidine-1-carboxylate

Take (S)-benzyl 2-((5-fluoro-2-(hydroxymethyl)-1H-benzo[D]imidazol-1-yl)methyl)pyrrolidine-1-carboxylate 20g (52.2 mmol) in a 500mL reaction flask, add 300mL of dichloromethane, under the protection of nitrogen, the temperature is reduced to 0℃, and 26.6g (62.6mmol) of Dess Martin reagent is added to it, returned to room temperature, stirred for 2 hours, TLC detects that the reaction is complete , Cooled to 0℃, washed with NaHCO ₃ /Na ₂ S ₂ O ₃ (1:1) saturated solution (200mL*2), the aqueous phase was extracted with dichloromethane (200mL*2), and the combined organic layer was washed with brine It was washed, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:5) to obtain (S)-benzyl 2-((5-fluoro-2-formyl-1H-benzo[D] (Imidazol-1-yl)methyl)pyrrolidine-1-carboxylate (18.3 g, 92%).

Example 7: (2S,2'S)-dibenzyl 2,2'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzolo[d]imidazole]-1 ,1'-Diyl)bis(methylene))bis(pyrrolidine-1-carboxylate) preparation

Take 15 g (39.4 mmol) of (S)-benzyl 2-((5-fluoro-2-formyl-1H-benzo[D]imidazol-1-yl)methyl)pyrrolidine-1-carboxylate, Benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]pyrrolidine-1-carboxylate 16.2g (47.2mmol) in a 250mL reaction flask, add 150mLN,N- Add 28.5g (0.15mol) of sodium metabisulfite, add 28.5g (0.15mol) of sodium metabisulfite, under the condition of microwave, reflux and stir for 1 hour, TLC detects that the reaction is complete, return to room temperature, add 500mL of water, and extract with ethyl acetate (200mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:5) to obtain (2S,2'S)-dibenzyl 2,2'-((5,5'-difluoro-1H, 1'H-[2,2'-Dibenzolo[d]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidine-1-carboxylate) (22.5g, 81 %).

Example 8: 5,5'-Difluoro-1,1'-bis((S)-pyrrolidin-2-ylmethyl)-1H,1'H-2,2'-dibenzo[D] Preparation of imidazole

Take (2S,2'S)-dibenzyl 2,2'-((5,5'-difluoro-1H,1'H-[2,2'-dibenz[d]imidazole]-1,1' -Diyl) bis(methylene)) bis(pyrrolidine-1-carboxylate) 10g (14.2mmol) in a 250mL reaction flask, add 100mL methanol, and another 1g palladium charcoal added to it, under a hydrogen atmosphere, room temperature After stirring for 12 hours, TLC detected that the reaction was complete, filtered, and the filtrate was concentrated to obtain 5,5'-difluoro-1,1'-bis((S)-pyrrolidin-2-ylmethyl)-1H,1'H- 2,2'-Dibenzo[D]imidazole (5.7g, 93% crude product) was directly used in the next reaction.

Example 9: Di-tert-butyl ((2R,2'R)-((2S,2'S)-2,2'-((5,5'-difluoro-1H,1'H-[2,2' -Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidine-2,1-diyl))bis(3-methyl-1-oxobutane) Alkyl-2,1-diyl)) Preparation of Dicarbamate

Take 2.2g (10mmol) of N-Boc-L valine in a 100mL reaction flask, add 20mL of N,N-dimethylformamide, and take N-methylmorpholine 2g (20mmol), HATU5.7g (15mmol) Add to it, stir at room temperature for 30 minutes, and then take 5,5'-difluoro-1,1'-bis((S)-pyrrolidin-2-ylmethyl)-1H,1'H-2,2'- 2g (4.6mmol) of dibenzo[D]imidazole was dissolved in 20mL of N,N-dimethylformamide, added dropwise to the above system, stirred at room temperature for 3 hours, TLC detected that the reaction was complete, 200mL of water, ethyl acetate (200mL× 3) Extraction, the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:3) to obtain di-tert-butyl ((2R,2'R)-((2S,2'S)-2,2'-( (5,5'-Difluoro-1H,1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidine- 2,1-diyl)) bis(3-methyl-1-oxobutane-2,1-diyl)) dicarbamate (2.5 g, 65%).

Example 10: (2R,2'R)-1,1'-((2S,2'S)-2,2'-((5,5'-difluoro-1H,1'H-[2,2' -Dibenzo[D]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidine-2,1-diyl))bis(2-amino-3-methylbutane -1-ketone) preparation

Take di-tert-butyl ((2R,2'R)-((2S,2'S)-2,2'-((5,5'-difluoro-1H,1'H-[2,2'-diphenyl And [d]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidine-2,1-diyl))bis(3-methyl-1-oxobutane-2 ,1-Diyl)) dicarbamate 1.0g (1.2mmol) in a 100mL reaction flask, add 20mL of hydrochloric acid ethanol solution, stir at room temperature for 1 hour, TLC detects the reaction is complete, the solvent and excess hydrogen chloride are evaporated under reduced pressure to obtain Colorless oily liquid 0.8g, directly used in the next reaction.

Example 11: Di-tert-butyl ((2R,2'R)-((2R,2'R)-((2S,2'S)-2,2'-((5,5'-difluoro-1H, 1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidine-2,1-diyl))bis(3 -Methyl-1-oxobutane-2,1-diyl))bis(azadiyl))bis(1-oxopropane-2,1-diyl))bis(methylcarbamate) )

Preparation

Take 0.58g (2.88mmol) of N-Boc-N-methyl-L alanine in a 100mL reaction flask, add 20mL of N,N-dimethylformamide, and take another 0.61g (6.0mmol) of N-methylmorpholine ), HATU1.37g (3.6mmol) was added to it, stirred at room temperature for 30 minutes, and then (2R,2'R)-1,1'-((2S,2'S)-2,2'-((5,5' -Difluoro-1H,1'H-[2,2'-Dibenzo[D]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidine-2,1-di (Base)) bis(2-amino-3-methylbutan-1-one) 0.8g (crude product, 1.2mmol) was dissolved in 20mL N,N-dimethylformamide, added dropwise to the above system, and stirred at room temperature for 3 hours , TLC detected that the reaction was complete, 200 mL of water was added, and ethyl acetate (200 mL×3) was added. The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:3) to obtain di-tert-butyl ((2R,2'R)-((2R,2'R)-((2S, 2'S)-2,2'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]-1,1'-diyl) bis( Methyl))bis(pyrrolidine-2,1-diyl))bis(3-methyl-1-oxobutane-2,1-diyl))bis(azadiyl))bis(1 -Oxopropane-2,1-diyl))bis(methylcarbamate) 0.68 g (53.5%).

Example 12: (2R,2'R)-N,N'-((2R,2'R)-((2S,2'S)-2,2'-((5,5'-difluoro-1H, 1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidine-2,1-diyl))bis(3 -Methyl-1-oxobutane-2,1-diyl)) bis(2-(methylamino)propionamide) preparation

Take the di-tert-butyl ((2R,2'R)-((2R,2'R)-((2S,2'S)-2,2'-((5,5'-difluoro-1H,1'H -[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidine-2,1-diyl))bis(3-methyl -1-oxobutane-2,1-diyl))bis(azadiyl))bis(1-oxopropane-2,1-diyl))bis(methylcarbamate) 0.68g (0.68mmol) In a 100mL reaction flask, add 20mL of hydrochloric acid ethanol solution, stir at room temperature for 1 hour, TLC detects that the reaction is complete, the solvent and excess hydrogen chloride are evaporated under reduced pressure, add 100mL of water, saturated sodium bicarbonate to adjust pH=8, ethyl acetate Ester extraction (100 mL*2), the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:2) to obtain (2R,2'R)-N,N'-((2R,2'R)-((2S, 2'S)-2,2'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]-1,1'-diyl) bis( Methyl))bis(pyrrolidine-2,1-diyl))bis(3-methyl-1-oxobutane-2,1-diyl))bis(2-(methylamino)propionamide) 312 mg (57%).

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.12-8.07 (s, 2H), 7.57-7.45 (m, 2H), 7.34-7.21 (m, 2H), 7.11-7.00 (m, 2H) ,4.62-4.54(d,2H),4.06-3.94(m,2H),3.86-3.71(m,2H),3.62-3.53(m,2H),3.44-3.31(m,6H),3.12-3.06( s,6H),2.75-2.66(m,2H),2.28-2.06(m,10H),1.33-1.24(m,6H),0.98-0.83(d,12H),(M+H) ⁺ :m/ z=805.6

With reference to the synthesis methods of Examples 1-12 and the first synthesis scheme, the following example compounds were synthesized:

Example 13:

¹ H-NMR(400MHz, d ₆ -DMSO): δ=8.15-8.09(s,2H), 7.61-7.48(m,2H), 7.33-7.25(m,2H), 7.14-7.02(m,2H) ,4.65-4.57(d,2H),4.11-3.98(m,2H),3.92-3.81(m,2H),3.70-3.63(m,2H),3.56-3.47(m,6H),3.34-3.18( s, 6H), 2.66-2.53 (m, 2H), 2.30-2.01 (m, 10H), 1.78-1.66 (m, 4H), 1.38-1.22 (m, 6H), 0.91-0.78 (d, 12H), [M+H] ⁺ :m/z=834.5

Example 14:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.10-8.02 (s, 2H), 7.51-7.42 (m, 2H), 7.36-7.27 (m, 2H), 7.10-6.93 (m, 2H) ,4.77-4.58(d,2H),4.26-4.11(m,2H),3.93-3.81(m,2H),3.78-3.66(m,2H),3.52-3.43(m,6H),3.18-3.02( s,6H),2.83-2.69(m,2H),2.37-2.12(m,10H),1.39-1.36(m,6H),1.15-0.98(d,6H),(M+H) ⁺ :m/ z=749.4

Example 15:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.20-8.14 (s, 2H), 7.66-7.51 (m, 2H), 7.43-7.36 (m, 2H), 7.18-7.05 (m, 2H) , 4.89-4.73(d,2H),4.58-4.41(m,2H),4.11-4.03(m,2H),3.87-3.72(m,6H),3.66-3.50(s,2H),3.41-3.33( m,6H),3.12-3.03(s,6H),2.87-2.65(m,2H),2.47-2.32(m,10H),1.59-1.42(m,6H),(M+H) ⁺ :m/ z=781.7

Example 16:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.22-8.19 (s, 2H), 7.62-7.48 (m, 2H), 7.32-7.25 (m, 2H), 7.16-7.04 (m, 2H) ,4.66-4.57(d,2H),4.01-3.86(m,2H),3.81-3.75(m,2H), 3.60-3.55(m,2H),3.42-3.33(m,6H),3.11-3.08( s,6H),2.77-2.69(m,2H),2.18-2.01(m,10H),1.45-1.34(m,6H),1.05-0.78(d,24H),(M+H) ⁺ :m/ z=834.6

Example 17:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.16-8.09 (s, 2H), 7.59-7.47 (m, 2H), 7.36-7.28 (m, 2H), 7.15-7.03 (m, 2H) ,4.68-4.55(d,2H),4.09-3.97(m,2H),3.88-3.73(m,2H),3.66-3.59(m,2H),3.41-3.30(m,6H),3.17-3.09( s,6H),2.77-2.69(m,2H),2.2 4-2.02(m,10H),1.38-1.26(m,6H),0.94-0.87(d,6H),(M+H) ⁺ :m /z=777.8

Example 18:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.11-8.05 (s, 2H), 7.52-7.44 (m, 2H), 7.36-7.26 (m, 2H), 7.13-7.01 (m, 2H) ,4.67-4.55(d,2H),4.08-3.95(m,2H),3.88-3.74(m,2H),3.65-3.51(m,2H),3.46-3.38(m,6H),3.15-3.09( s,6H),2.72-2.65(m,2H),2.23-2.05(m,10H),1.63-1.54(m,6H),1.36-1.22(m,6H),0.99-0.86(d,6H), [M+H] ⁺ :m/z=834.3

Example 19:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.18-8.10 (s, 2H), 7.75-7.59 (m, 2H), 7.49-7.38 (m, 2H), 7.22-7.09 (m, 2H) , 4.95-4.81(d, 2H), 4.52-4.46(m, 2H), 4.17-4.08(m, 2H), 3.88-3.75(m, 6H), 3.63-3.51(s, 2H), 3.45-3.32( m,6H),3.11-3.07(s,6H),2.81-2.68(m,2H),2.43-2.36(m,10H),1.52-1.45(m,6H),1.24-1.15(m,6H)[ M+H] ⁺ :m/z＝810.2

Example 20:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.11-8.05 (s, 2H), 7.53-7.45 (m, 2H), 7.32-7.25 (m, 2H), 7.15-6.96 (m, 2H) ,4.73-4.54(d,2H),4.26-4.14(m,2H),3.96-3.83(m,2H),3.73-3.65(m,2H),3.53-3.45(m,6H),3.13-3.05( s, 6H), 2.85-2.67 (m, 2H), 2.34-2.15 (m, 10H), 1.37-1.30 (m, 6H), 1.25-0.92 (d, 2H), 0.52-0.35 (m, 8H), [M+H] ⁺ :m/z=801.4

Example 21:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.13-8.08 (s, 2H), 7.51-7.43 (m, 2H), 7.35-7.21 (m, 2H), 7.11-6.92 (m, 2H) ,4.77-4.56(d,2H),4.22-4.11(m,2H),3.98-3.79(m,2H),3.71-3.62(m,2H),3.54-3.44(m,6H),3.11-3.08( s, 6H), 2.88-2.66 (m, 2H), 2.58-2.45 (m, 2H), 2.33-2.18 (m, 10H), 1.73-1.66 (d, 6H), 1.42-1.26 (m, 20H), [M+H] ⁺ :m/z=886.3

Example 22:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.22-8.18 (s, 2H), 7.78-7.66 (m, 2H), 7.53-7.47 (m, 2H), 7.39-7.28 (m, 4H) ,7.20-7.13(m,4H),7.05-6.92(m,4H),4.83-4.71(d,2H), 4.60-4.47(m,2H),4.12-4.05(m,2H),3.82-3.71( m, 6H), 3.69-3.53 (s, 2H), 3.46-3.34 (m, 6H), 3.18-3.04 (s, 6H), 2.85-2.62 (m, 2H), 2.44-2.31 (m, 10H), 1.54-1.47(m,6H),[M+H] ⁺ :m/z=902.7

Example 23:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.26-8.19 (s, 2H), 7.83-7.67 (m, 2H), 7.57-7.49 (m, 2H), 7.34-7.22 (m, 4H) ,7.18-7.10(m,4H),7.02-6.87(m,2H),5.23-5.15(s,2H),4.87-4.73(d,2H),4.63-4.49(m,2H),4.15-4.09( m,2H),3.87-3.78(m,6H),3.63-3.52(s,2H),3.45-3.33(m,6H),3.14-3.00(s,6H),2.89-2.66(m,2H), 2.47-2.32(m,10H),1.55-1.42(m,6H),[M+H] ⁺ :m/z=934.2

Example 24:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=9.92-9.84 (s, 2H), 8.18-8.10 (s, 2H), 7.73-7.61 (m, 2H), 7.50-7.40 (m, 2H) ,7.32-7.24(m,4H),7.18-7.08(m,4H),7.00-6.83(m,4H),4.88-4.75(d,2H), 4.67-4.49(m,2H),4.15-4.08( m, 2H), 3.85-3.74 (m, 6H), 3.68-3.54 (s, 2H), 3.42-3.32 (m, 6H), 3.16-3.05 (s, 6H), 2.88-2.64 (m, 2H), 2.48-2.37(m,10H),1.56-1.49(m,6H),[M+H] ⁺ :m/z=980.7

Example 25:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 11.76-11.64 (s, 2H), 8.57-8.42 (s, 2H), 8.20-8.17 (s, 2H), 7.83-7.69 (m, 2H) ,7.55-7.43(m,2H),7.38-7.26(m,2H),7.17-7.04(m,2H),4.76-4.65(d,2H),4.57-4.41(m,2H),4.19-4.05( m,2H),3.83-3.72(m,6H),3.65-3.51(s,2H),3.46-3.33(m,6H),3.14-3.06(s,6H),2.85-2.62(m,2H), 2.43-2.34(m,10H),1.54-1.44(m,6H),[M+H] ⁺ :m/z=882.1

Example 26: (S)-Benzyl 2-((1'-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)methyl)-5,5'-difluoro- Preparation of 1H,1'H-[2,2'-bibenzo[d]imidazole]-1-yl)methyl)pyrrolidine-1-carboxylate

Take 15 g (39.4 mmol) of (S)-benzyl 2-((5-fluoro-2-formyl-1H-benzo[D]imidazol-1-yl)methyl)pyrrolidine-1-carboxylate, Tert-butoxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]pyrrolidine-1-carboxylate 14.5g (47.2mmol) in a 250mL reaction flask, add 150mLN,N -Dimethylformamide, add 28.5g (0.15mol) sodium metabisulfite, under nitrogen protection, under microwave conditions, reflux and stir for 1 hour, TLC detects that the reaction is complete, return to room temperature, add 500mL water, and extract with ethyl acetate (200mL×3) The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:5) to obtain (S)-benzyl 2-((1'-((S)-1-(tert-butoxycarbonyl) )Pyrrolidin-2-yl)methyl)-5,5'-difluoro-1H,1'H-[2,2'-bibenzo[d]imidazole]-1-yl)methyl)pyrrolidine -1-carboxylate (20.6 g, 78%).

Example 27: (S)-Benzyl 2-((5,5'-difluoro-1'-((S)-pyrrolidin-2-ylmethyl)-1H,1'H-[2,2 Preparation of'-bibenzo[d]imidazole]-1-yl)methyl)pyrrolidine-1-carboxylate

Take (S)-benzyl 2-((1'-((S)-1-(tert-butoxycarbonyl)pyrrolidin-2-yl)methyl)-5,5'-difluoro-1H,1 10g (14.9mmol) of'H-[2,2'-bibenzo[d]imidazole]-1-yl)methyl)pyrrolidine-1-carboxylate is placed in a 250mL reaction flask, and 120mL of hydrochloric acid ethanol solution is added, Stir at room temperature for 1 hour. TLC detects that the reaction is complete. The solvent and excess hydrogen chloride are evaporated under reduced pressure to obtain 8.6 g of a colorless oily liquid, which is directly used in the next reaction. , Directly used in the next reaction.

Example 28: (S)-Benzyl 2-((1'-((S)-1-((R)-2-((tert-butoxycarbonyl)amino)-3-methylbutyryl)pyrrolidine -2-yl)methyl)-5,5'-difluoro-1H,1'H-[2,2'-dibenzo[D]imidazole]-1-yl)methyl)pyrrolidine-1- Preparation of carboxylic acid ester

Take 3.9g (17.9mmol) of N-Boc-L valine in a 250mL reaction flask, add 80mL of N,N-dimethylformamide, and take another 4.5g (44.7mmol) of N-methylmorpholine, HATU8.5g (22.4mmol) was added to it, stirred at room temperature for 30 minutes, and then (S)-benzyl 2-((5,5'-difluoro-1'-((S)-pyrrolidin-2-ylmethyl)- 1H,1'H-[2,2'-Bibenzo[d]imidazole]-1-yl)methyl)pyrrolidine-1-carboxylate 8.6g (crude product, 14.9mmol) dissolved in 40mL N,N- Dimethylformamide was added dropwise to the above system, stirred at room temperature for 3 hours, TLC detected that the reaction was complete, 600mL of water was added, ethyl acetate (200mL×3) was added, the combined organic layer was washed with brine, and dried over anhydrous sodium sulfate , And concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:3) to obtain (S)-benzyl 2-((1'-((S)-1-((R)-2) -((Tert-Butoxycarbonyl)amino)-3-methylbutyryl)pyrrolidin-2-yl)methyl)-5,5'-difluoro-1H,1'H-[2,2'-di Benzo[D]imidazol]-1-yl)methyl)pyrrolidine-1-carboxylate 8.1 g (70%).

Example 29: tert-butyl((R)-1-((S)-2-((5,5'-difluoro-1'-((S)-pyrrolidin-2-ylmethyl)-1H ,1'H-[2,2'-Dibenzo[d]imidazole]-1-yl)methyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl ) Preparation of carbamate

Take (S)-benzyl 2-((1'-((S)-1-((R)-2-((tert-butoxycarbonyl)amino)-3-methylbutyryl)pyrrolidine-2- (Yl)methyl)-5,5'-difluoro-1H,1'H-[2,2'-dibenzo[D]imidazole]-1-yl)methyl)pyrrolidine-1-carboxylate 8.1g (10.5mmol) was put into a 250mL reaction flask, 100mL methanol was added, and 0.8g palladium charcoal was added to it. Under a hydrogen atmosphere, stirred at room temperature for 12 hours, TLC detected that the reaction was complete, filtered, and the filtrate was concentrated to obtain tert-butyl (( R)-1-((S)-2-((5,5'-Difluoro-1'-((S)-pyrrolidin-2-ylmethyl)-1H,1'H-[2,2 '-Dibenzo[d]imidazole]-1-yl)methyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)carbamate 6.7g crude product, Used directly in the next reaction.

Example 30: Di-tert-butyl((2R,2'R)-2-amino-1-((S)-2-((1'-((S)-1-((R)-2-amino (Propanol group)pyrrolidin-2-yl)methyl)-5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazol]-1-yl)methyl )Pyrrolidin-1-yl)-3-methylbutan-1-one-bis(methylcarbamate) preparation

Take 0.35g (1.89mmol) of N-Boc-D alanine in a 100mL reaction flask, add 20mL of N,N-dimethylformamide, and another 0.32g (3.14mmol) of N-methylmorpholine, 0.90g of HATU (2.36mmol) was added to it, stirred at room temperature for 30 minutes, and then tert-butyl ((R)-1-((S)-2-((5,5'-difluoro-1'-((S)-pyrrole)) Alkyl-2-ylmethyl)-1H,1'H-[2,2'-Dibenzo[d]imidazol]-1-yl)methyl)pyrrolidin-1-yl)-3-methyl- 1.0g (crude product, 1.57mmol) of 1-oxobutan-2-yl) carbamate was dissolved in 20mL of N,N-dimethylformamide, added dropwise to the above system, stirred at room temperature for 3 hours, TLC detected that the reaction was complete After adding 600 mL of water, extracting with ethyl acetate (100 mL×3), the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:5) to obtain di-tert-butyl ((2R, 2'R)-2-amino-1-((S)-2- ((1'-((S)-1-((R)-2-aminopropanol)pyrrolidin-2-yl)methyl)-5,5'-difluoro-1H,1'H-[ 2,2'-Dibenzo[d]imidazol]-1-yl)methyl)pyrrolidin-1-yl)-3-methylbutan-1-one-bis(methylcarbamate) 0.91g (72%).

Example 31: (R)-2-amino-1-((S)-2-((1'-((S)-1-((R)-2-aminopropanol))pyrrolidine-2- (Yl)methyl)-5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazol]-1-yl)methyl)pyrrolidin-1-yl)- Preparation of 3-methylbutan-1-one

Take di-tert-butyl ((2R,2'R)-2-amino-1-((S)-2-((1'-((S)-1-((R)-2-aminopropanol) )Pyrrolidin-2-yl)methyl)-5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]-1-yl)methyl)pyrrolidine -1-yl)-3-methylbutan-1-one-bis(methylcarbamate) 0.91g (1.1mmol) in a 100mL reaction flask, add 20mL hydrochloric acid ethanol solution, stir at room temperature for 1 hour, TLC detection After the reaction was complete, the solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 0.7 g of a colorless oily liquid, which was directly used in the next reaction.

Example 32: Di-tert-butyl ((2R,2'R)-(R)-N-((R)-1-((S)-2-((5,5'-difluoro-1'- ((S)-1-((R)-2-((R)-2-(methylamino)alanyl)pyrrolidin-2-yl)methyl)-1H,1'H-[2, 2'-Bibenzo[D]imidazole]-1-yl)methyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-(methylamino) ) Preparation of propionamide-bis(methylcarbamate)

Take 0.54g (2.64mmol) of N-Boc-N-methyl-L alanine in a 100mL reaction flask, add 20mL of N,N-dimethylformamide, and take 0.56g (5.5mmol of N-methylmorpholine) ), HATU1.3g (3.3mmol) was added to it, stirred at room temperature for 30 minutes, and then (R)-2-amino-1-((S)-2-((1'-((S)-1-(() R)-2-Aminopropanol)pyrrolidin-2-yl)methyl)-5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]- 1-yl)methyl)pyrrolidin-1-yl)-3-methylbutan-1-one 0.7g (1.1mmol) dissolved in 20mL N,N-dimethylformamide, added dropwise to the above system, room temperature After stirring for 3 hours, TLC detected that the reaction was complete, 200 mL of water was added, and extraction was performed with ethyl acetate (200 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:3) to obtain di-tert-butyl ((2R, 2'R)-(R)-N-((R)-1- ((S)-2-((5,5'-Difluoro-1'-((S)-1-((R)-2-((R)-2-(methylamino)alanyl) (Pyrrolidin-2-yl)methyl)-1H,1'H-[2,2'-Bibenzo[D]imidazole]-1-yl)methyl)pyrrolidin-1-yl)-3-methyl -1-oxobutan-2-yl)-2-(methylamino)propionamide-bis(methylcarbamate) 0.66 g (61%).

Example 33: (R)-N-((R)-1-((S)-2-((5,5'-difluoro-1'-((S)-1-((R)-2 -((R)-2-(Methylamino)alanyl)pyrrolidin-2-yl)methyl)-1H,1'H-[2,2'-Bibenzo[D]imidazole]-1 -Yl)methyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-(methylamino)propionamide

Take the di-tert-butyl ((2R,2'R)-(R)-N-((R)-1-((S)-2-((5,5'-difluoro-1'-((S )-1-((R)-2-((R)-2-(methylamino)alanyl)pyrrolidin-2-yl)methyl)-1H,1'H-[2,2'- Bibenzo [D]imidazole]-1-yl)methyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-(methylamino)propionamide -0.66g (0.68mmol) of bis(methylcarbamate) was added to a 100mL reaction flask, 20mL of hydrochloric acid ethanol solution was added, stirred at room temperature for 1 hour, TLC detected that the reaction was complete, the solvent and excess hydrogen chloride were evaporated under reduced pressure, and 100mL of water was added. Saturated sodium bicarbonate adjusted pH=8, extracted with ethyl acetate (100mL*2), the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: Ethyl acetate: petroleum ether = 1:2) to obtain (R)-N-((R)-1-((S)-2-((5,5'-difluoro-1'-((S) -1-((R)-2-((R)-2-(methylamino)alanyl)pyrrolidin-2-yl)methyl)-1H,1'H-[2,2'-ratio Benzo[D]imidazole]-1-yl)methyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)-2-(methylamino)propionamide 513mg (62%).

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.17-8.09 (s, 2H), 7.67-7.55 (m, 2H), 7.42-7.31 (m, 2H), 7.18-7.05 (m, 2H) ,4.68-4.53(d,2H),4.11-3.98(m,2H),3.88-3.74(m,2H),3.66-3.54(m,2H),3.48-3.34(m,6H),3.16-3.09( s, 6H), 2.85-2.75 (m, 2H), 2.33-2.16 (m, 10H), 1.65-1.51 (m, 6H), 1.38-1.23 (d, 3H), 0.99-0.82 (d, 6H), [M+H] ⁺ :m/z=777.8

With reference to the synthetic methods of Examples 26-33 and the second synthetic scheme, the following example compounds were synthesized:

Example 34:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.11-8.02 (s, 2H), 7.77-7.62 (m, 2H), 7.52-7.38 (m, 2H), 7.28-7.15 (m, 2H) ,4.72-4.59(d,2H),4.35-4.17(m,2H),3.81-3.72(m,2H),3.62-3.51(m,2H),3.44-3.35(m,6H),3.18-3.02( s, 6H), 2.87-2.73 (m, 2H), 2.38-2.13 (m, 10H), 1.63-1.53 (m, 6H), 1.34-1.22 (d, 2H), 0.94-0.71 (d, 12H), [M+H] ⁺ :m/z=820.2

Example 35:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.12-8.01(s,2H), 7.62-7.50(m,2H),7.43-7.36(m,2H),7.22-7.08(m,2H) ,4.78-4.64(d,2H),4.32-4.12(m,2H),3.94-3.81(m,2H),3.73-3.64(m,2H),3.55-3.47(m,6H),3.24-3.15( s, 6H), 2.98-2.85 (m, 6H), 2.45-2.36 (m, 10H), 2.07-1.92 (d, 2H), 1.36-1.20 (d, 3H), 1.06-0.89 (d, 6H), [M+H] ⁺ :m/z=794.1

Example 36:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.21-8.12 (s, 2H), 7.72-7.64 (m, 2H), 7.52-7.38 (m, 2H), 7.12-7.00 (m, 2H) ,4.83-4.63(d,2H),4.26-4.11(m,2H),3.83-3.71(m,2H),3.59-3.47(m,2H),3.38-3.24(m,6H),3.13-3.02( s, 6H), 2.77-2.65 (m, 2H), 2.43-2.26 (m, 10H), 1.75-1.59 (m, 6H), 1.33-1.20 (s, 9H), 0.93-0.84 (d, 6H), [M+H] ⁺ :m/z=820.3

Example 37:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.23-8.17 (s, 2H), 7.87-7.72 (m, 2H), 7.52-7.39 (m, 2H), 7.28-7.12 (m, 2H) , 4.87-4.65 (d, 2H), 4.27-4.12 (m, 2H), 3.99-3.86 (m, 2H), 3.77-3.64 (m, 2H), 3.53-3.46 (m, 6H), 3.28-3.16 ( s, 6H), 2.97-2.80 (m, 2H), 2.48-2.36 (m, 10H), 1.78-1.63 (m, 6H), 1.44-1.37 (m, 1H), 1.08-0.89 (d, 6H), 0.62-0.54(m,4H),[M+H] ⁺ :m/z＝804.1

Example 38:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.11-8.01 (s, 2H), 7.63-7.51 (m, 2H), 7.46-7.33 (m, 2H), 7.14-7.02 (m, 2H) ,4.64-4.52(d,2H),4.17-3.99(m,2H),3.83-3.71(m,2H),3.67-3.55(m,2H),3.45-3.32(m,6H),3.19-3.11( s, 6H), 2.82-2.73 (m, 2H), 2.38-2.19 (m, 10H), 2.03-1.86 (m, 10H), 1.62-1.50 (m, 6H), 1.33-1.21 (d, 1H), 0.93-0.80(d,6H),[M+H] ⁺ :m/z=846.4

Example 39:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.27-8.15 (s, 2H), 7.77-7.65 (m, 2H), 7.48-7.34 (m, 3H), 7.27-7.20 (2H), 7.16 -7.07(m,4H),4.73-4.68(d,2H),4.35-4.18(m,2H),3.93-3.84(m,2H),3.73-3.59(m,2H),3.42-3.30(m, 6H), 3.14-3.06 (s, 6H), 2.87-2.72 (m, 2H), 2.36-2.19 (m, 10H), 1.64-1.54 (m, 6H), 1.34-1.22 (d, 1H), 0.89- 0.73(d,6H),[M+H] ⁺ :m/z=854.4

Example 40:

¹ H-NMR(400MHz,d ₆ -DMSO): δ=9.83-9.71(d,1H), 8.12-8.02(s,2H), 7.62-7.52(m,3H),7.44-7.33(m,4H) ,7.15-7.04(m,4H),4.62-4.51(d,2H),4.17-3.99(m,2H),3.82-3.71(m,2H),3.64-3.51(m,2H),3.44-3.31( m,6H),3.13-3.02(s,6H),2.89-2.78(m,2H),2.36-2.18(m,10H),1.64-1.53(m,6H),1.35-1.28(d,1H), 0.92-0.81(d,6H),[M+H] ⁺ :m/z=893.5

Example 41:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 12.87-12.73 (s, 1H), 8.93-8.85 (d, 2H), 8.22-8.16 (s, 2H), 7.66-7.58 (m, 2H) ,7.49-7.37(m,2H),7.13-7.02(m,2H),4.77-4.65(d,2H),4.21-4.06(m,2H),3.89-3.76(m,2H),3.68-3.57( m,2H),3.49-3.37(m,6H),3.26-3.14(s,6H),2.88-2.76(m,2H),2.38-2.22(m,10H),1.77-1.58(m,6H), 1.34-1.26(d,1H),0.94-0.86(d,6H),[M+H] ⁺ :m/z=844.1

Example 42: Preparation of benzyloxycarbonyl (2S)-2-[(1,3-dioxoisoindol-2-yl)methyl]-4-hydroxypyrrolidine-1-carboxylate

Take 80g (0.319mol) of N-Cbz-4-hydroxy-L-prolinol, 56.2g (0.382mol) of phthalimide and 83.6g (0.319mol) of triphenylphosphine in a 2L three-necked reaction flask , Adding tetrahydrofuran (1.3L), cooling the system to 0°C under the protection of nitrogen, and adding DIAD 64.4g (0.319mol) dropwise at 0°C. Then the system was heated to room temperature and stirred for 16 hours. TLC detected that the reaction was complete. 400 mL of water was added for quenching, stirring was continued for 30 minutes, and extracted with ethyl acetate (300 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:20) to obtain benzyloxycarbonyl (2S)-2-[(1,3-dioxoisoindol-2-yl) Methyl]-4-hydroxypyrrolidine-1-carboxylate (64.2 g, 53%).

Example 43: Preparation of benzyloxycarbonyl (2S)-2-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylate

Take 64.2g (0.17mol) of benzyloxycarbonyl(2S)-2-[(1,3-dioxoisoindol-2-yl)methyl]-4-hydroxypyrrolidine-1-carboxylate in 1L Into the reaction flask, add 450 mL of ethanol, heat to 80°C, and add 17 g (0.45 mol, 85%) of hydrazine hydrate to it dropwise. Then it was stirred at 80°C for 12 hours. TLC detected that the reaction was complete. Cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain benzyloxycarbonyl (2S)-2-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylate (38.3g, 90%), which was used directly for the next One step response.

Example 44: Preparation of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-nitro-anilino)methyl]-4-hydroxypyrrolidine-1-carboxylate

Take 37.5 g (0.15 mol) of benzyloxycarbonyl (2S)-2-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylate in a 500 mL reaction flask, add 350 mL of acetonitrile, and take 41.4 g of potassium carbonate. 0.3 mol) was added to it, and 26.2 g (0.17 mol) of 1,4-difluoro-2-nitrobenzene was added to it. Under the protection of nitrogen, the temperature was raised to 80°C and stirred for 1 hour. TLC detected that the reaction was complete, quenched by adding 200 mL of water, and extracted with ethyl acetate (200 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:20) to obtain benzyloxycarbonyl (2S)-2-[(4-fluoro-2-nitro-anilino)methyl] -4-Hydroxypyrrolidine-1-carboxylate (46.7 g, 80%).

Example 45: Preparation of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]-4-hydroxypyrrolidine-1-carboxylate

Take 45g (0.12mol) of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-nitro-anilino)methyl]-4-hydroxypyrrolidine-1-carboxylate in a 1L reaction flask, Add 400mL of acetic acid, add another 135g of iron powder to it, stir at room temperature for 16 hours, TLC test the reaction is complete, filter, the filter cake is rinsed with ethyl acetate, the filtrate is concentrated under reduced pressure, 300mL of water, ethyl acetate (200mL×3) After extraction, the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Obtained benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]-4-hydroxypyrrolidine-1-carboxylate (38.6g, 93%), which was used directly for the next One step response.

Example 46: (2S)-Benzyl 2-((5-fluoro-2-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)-4-hydroxypyrrolidine-1 -Preparation of carboxylic acid ester

Take 31.3g (87.4mmol) of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]-4-hydroxypyrrolidine-1-carboxylate in a 500mL reaction flask, Add 300mL of 4N hydrochloric acid, and add 20g (0.262mol) of acetic alcohol to it. Heat to 100°C and stir for 6 hours. TLC detects that the reaction is complete. Cool down to 0°C. Add saturated sodium bicarbonate solution to adjust pH=8-9, filter, The filter cake was washed with methyl tert-butyl ether and dried to obtain (2S)-benzyl 2-((5-fluoro-2-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl )-4-hydroxypyrrolidine-1-carboxylate (28.9g, 83%), used directly in the next reaction.

Example 47: (2S)-Benzyl 2-((5-fluoro-2-formyl-1H-benzo[d]imidazol-1-yl)methyl)-4-hydroxypyrrolidine-1-carboxylic acid Preparation of salt ester

Take (2S)-benzyl 2-((5-fluoro-2-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)-4-hydroxypyrrolidine-1-carboxylic acid Put 20.8g (52.2mmol) of ester in a 500mL reaction flask, add 300mL of dichloromethane, under the protection of nitrogen, cool to 0℃, and add 26.6g (62.6mmol) of Des Martin reagent to it, return to room temperature, and stir for 2 hours , TLC detects that the reaction is complete, cool to 0°C, _{wash with saturated NaHCO 3} /Na ₂ S ₂ O ₃ (1:1) solution (200 mL*2), extract the aqueous phase with dichloromethane (200 mL*2), and combine The organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:5) to obtain (2S)-benzyl 2-((5-fluoro-2-formyl-1H-benzo[d] (Imidazol-1-yl)methyl)-4-hydroxypyrrolidine-1-carboxylate (18.7 g, 90%).

Example 48: (5S,5'S)-dibenzyl 5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenz[d]imidazole]-1 ,1'-Diyl)bis(methylene))bis(3-hydroxypyrrolidine-1-carboxylate)

Take (2S)-benzyl 2-((5-fluoro-2-formyl-1H-benzo[d]imidazol-1-yl)methyl)-4-hydroxypyrrolidine-1-carboxylate 15g (37.8mmol), benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]-4-hydroxypyrrolidine-1-carboxylate 16.3g (45.3mmol) in 250mL Into the reaction flask, add 150mL of N,N-dimethylformamide, add 28.5g (0.15mol) of sodium metabisulfite, under nitrogen protection, reflux and stir for 1 hour under microwave conditions, TLC detects the reaction is complete, return to room temperature, add 500mL of water, acetic acid Ethyl acetate (200 mL×3) was extracted, and the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:5) to obtain (5S,5'S)-dibenzyl 5,5'-((5,5'-difluoro-1H, 1'H-[2,2'-Dibenzyl[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-hydroxypyrrolidine-1-carboxylate) (21.7 g, 78%).

Example 49: (5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]-1,1' -Diyl)bis(methylene))bis(pyrrolidin-3-ol) preparation

Take (5S,5'S)-dibenzyl 5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzyl[d]imidazole]-1,1' -Diyl)bis(methylene))bis(3-hydroxypyrrolidine-1-carboxylate) 20g (27.2mmol) in a 250mL reaction flask, add 100mL methanol, and another 1g palladium charcoal added to it, hydrogen atmosphere After stirring at room temperature for 12 hours, TLC detected that the reaction was complete, filtered, and the filtrate was concentrated to obtain (5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2' -Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidin-3-ol) (11.6g, 91% crude product), directly used in the next reaction.

Example 50: Di-tert-butyl ((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2' -Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-hydroxypyrrolidine-5,1-diyl))bis(3-methyl-1- Preparation of oxobutane-2,1-diyl)) dicarbamate

Take 2.2g (10.3mmol) of N-Boc-L valine in a 100mL reaction flask, add 20mL of N,N-dimethylformamide, and take N-methylmorpholine 2.2g (21.5mmol), HATU4.9g (12.9mmol) was added to it, stirred at room temperature for 30 minutes, and then (5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzo [d]Imidazole]-1,1'-diyl)bis(methylene))bis(pyrrolidin-3-ol) 2g (4.3mmol) dissolved in 20mL N,N-dimethylformamide, add dropwise to the above The system was stirred at room temperature for 3 hours. TLC detected that the reaction was complete. 200 mL of water was added and extracted with ethyl acetate (200 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:3) to obtain di-tert-butyl ((2R,2'R)-((5S,5'S)-5,5'-( (5,5'-Difluoro-1H,1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-hydroxy Pyrrolidine-5,1-diyl))bis(3-methyl-1-oxobutane-2,1-diyl))dicarbamate (2.2 g, 58%).

Example 51: (2R,2'R)-1,1'-((5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2' -Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-hydroxypyrrolidine-5,1-diyl))bis(2-amino-3-methyl Butan-1-one)

Preparation

Take di-tert-butyl ((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2'-diphenyl And [d]imidazole]-1,1'-diyl)bis(methylene))bis(3-hydroxypyrrolidine-5,1-diyl))bis(3-methyl-1-oxobutan) Alkane-2,1-diyl)) dicarbamate 1.0g (1.2mmol) was added to a 100mL reaction flask, 20mL of hydrochloric acid ethanol solution was added, and the mixture was stirred at room temperature for 1 hour. TLC detected that the reaction was complete, and the solvent and excess were evaporated under reduced pressure. Hydrogen chloride, 0.8 g of colorless oily liquid was obtained, which was directly used in the next reaction.

Example 52: Di-tert-butyl ((2R,2'R)-((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H, 1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-hydroxypyrrolidine-5,1-diyl)) Bis(3-methyl-1-oxobutane-2,1-diyl))bis(azadiyl))bis(1-oxopropane-2,1-diyl))bis(methylamino) Preparation of methyl formate)

Take 0.58g (2.88mmol) of N-Boc-N-methyl-L alanine in a 100mL reaction flask, add 20mL of N,N-dimethylformamide, and take another 0.61g (6.0mmol) of N-methylmorpholine ), HATU1.37g (3.6mmol) was added to it, stirred at room temperature for 30 minutes, and then (2R,2'R)-1,1'-((5S,5'S)-5,5'-((5,5' -Difluoro-1H,1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-hydroxypyrrolidine-5, 1-Diyl)) bis(2-amino-3-methylbutane-1-one) 0.8g (crude product, 1.2mmol) dissolved in 20mL N,N-dimethylformamide, added dropwise to the above system, room temperature After stirring for 3 hours, TLC detected that the reaction was complete, 200 mL of water was added, and the mixture was extracted with ethyl acetate (200 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:3) to obtain di-tert-butyl ((2R, 2'R)-((2R, 2'R)-((5S, 5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]-1,1'-diyl) bis( Methyl))bis(3-hydroxypyrrolidine-5,1-diyl))bis(3-methyl-1-oxobutane-2,1-diyl))bis(azadiyl)) Bis(1-oxopropane-2,1-diyl))bis(methylcarbamate) 0.57 g (46%).

Example 53: (2R,2'R)-N,N'-((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H, 1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-hydroxypyrrolidine-5,1-diyl)) Preparation of bis(3-methyl-1-oxobutane-2,1-diyl))bis(2-(methylamino)propionamide)

Take the di-tert-butyl ((2R,2'R)-((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H -[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-hydroxypyrrolidine-5,1-diyl))bis(3 -Methyl-1-oxobutane-2,1-diyl))bis(azadiyl))bis(1-oxopropane-2,1-diyl))bis(methylcarbamate) ) 0.57g (0.55mmol) in a 100mL reaction flask, add 20mL of hydrochloric acid ethanol solution, stir at room temperature for 1 hour, TLC detects that the reaction is complete, the solvent and excess hydrogen chloride are evaporated under reduced pressure, 100mL of water is added, saturated sodium bicarbonate is adjusted to pH=8 , Ethyl acetate extraction (100 mL*2), the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:2) to obtain (2R,2'R)-N,N'-((2R,2'R)-((5S, 5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]-1,1'-diyl) bis( Methyl))bis(3-hydroxypyrrolidine-5,1-diyl))bis(3-methyl-1-oxobutane-2,1-diyl))bis(2-(methylamino) Propionamide) 300 mg (65%).

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.10-8.02 (s, 2H), 7.53-7.41 (m, 2H), 7.33-7.24 (m, 2H), 7.16-7.04 (m, 2H) ,4.66-4.57(d,2H),4.16-3.99(m,2H),3.83-3.74(m,2H),3.66-3.52(m,2H),3.42-3.35(m,6H),3.16-3.09( s,6H),2.85-2.73(m,2H),2.48-2.26(m,10H),1.45-1.34(m,6H),0.99-0.86(d,12H),(M+H) ⁺ :m/ z=837.8

Referring to the synthesis methods of Examples 42-53 and the first synthesis scheme, the following example compounds were synthesized:

Example 54:

¹ H-NMR(400MHz, d ₆ -DMSO): δ=8.16-8.05(s,2H), 7.55-7.46(m,2H), 7.39-7.27(m,2H), 7.16-6.95(m,2H) ,4.78-4.59(d,2H),4.25-4.14(m,2H),3.97-3.84(m,2H),3.77-3.63(m,2H),3.54-3.42(m,6H),3.16-3.05( s,6H),2.88-2.72(m,2H),2.32-2.11(m,10H),1.34-1.22(m,6H),1.14-0.96(d,6H),(M+H) ⁺ :m/ z=781.6

Example 55:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.19-8.11 (s, 2H), 7.62-7.49 (m, 2H), 7.33-7.23 (m, 2H), 7.13-7.02 (m, 2H) ,4.64-4.52(d,2H),4.01-3.92(m,2H),3.84-3.72(m,2H),3.68-3.57(m,2H),3.43-3.33(m,6H),3.16-3.07( s,6H),2.74-2.65(m,2H),2.26-2.08(m,10H),1.37-1.24(m,6H),0.97-0.88(d,6H),(M+H) ⁺ :m/ z=810.1

Example 56:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.17-8.06 (s, 2H), 7.58-7.49 (m, 2H), 7.34-7.25 (m, 2H), 7.19-7.04 (m, 2H) ,4.65-4.52(d,2H),4.04-3.92(m,2H),3.85-3.73(m,2H),3.68-3.55(m,2H),3.42-3.33(m,6H),3.17-3.08( s,6H),2.77-2.68(m,2H),2.26-2.09(m,10H),1.65-1.57(m,6H),1.33-1.27(m,6H),0.92-0.84(d,6H), [M+H] ⁺ :m/z＝866.3

Example 57:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.21-8.14 (s, 2H), 7.64-7.55 (m, 2H), 7.38-7.27 (m, 2H), 7.13-7.02 (m, 2H) ,4.69-4.56(d,2H),4.03-3.94(m,2H),3.85-3.78(m,2H),3.65-3.57(m,2H),3.44-3.37(m,6H),3.18-3.09( s,6H),2.73-2.62(m,2H),2.13-2.06(m,10H),1.46-1.38(m,6H),1.01-0.76(d,24H),(M+H) ⁺ :m/ z=866.6

Example 58:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.16-8.08(s,2H), 7.63-7.55(m,2H),7.45-7.33(m,2H), 7.17-7.04(m,2H) ,4.86-4.74(d,2H),4.55-4.43(m,2H),4.18-4.09(m,2H),3.83-3.75(m,6H),3.63-3.57(s,2H),3.44-3.38( m,6H),3.13-3.06(s,6H),2.88-2.63(m,2H),2.49-2.37(m,10H),1.56-1.43(m,6H),(M+H) ⁺ :m/ z=813.7

Example 59:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.11-8.02 (s, 2H), 7.73-7.56 (m, 2H), 7.42-7.31 (m, 2H), 7.24-7.11 (m, 2H) , 4.87-4.76(d,2H),4.56-4.48(m,2H),4.13-4.02(m,2H),3.89-3.73(m,6H),3.66-3.54(s,2H),3.42-3.36( m,6H),3.14-3.08(s,6H),2.86-2.63(m,2H),2.48-2.39(m,10H),1.54-1.43(m,6H),1.26-1.18(m,6H)[ M+H] ⁺ :m/z=841.8

Example 60:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.22-8.13 (s, 2H), 7.64-7.47 (m, 2H), 7.38-7.24 (m, 2H), 7.12-7.04 (m, 2H) ,4.67-4.59(d,2H),4.18-4.09(m,2H),3.90-3.84(m,2H),3.75-3.66(m,2H),3.53-3.45(m,6H),3.36-3.19( s,6H),2.62-2.51(m,2H),2.36-2.21(m,10H),1.74-1.65(m,4H),1.32-1.21(m,6H),0.98-0.86(d,12H), [M+H] ⁺ :m/z＝866.2

Example 61:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.12-8.03(s, 2H), 7.54-7.41(m, 2H), 7.33-7.25(m, 2H), 7.12-6.97(m, 2H) ,4.75-4.57(d,2H),4.23-4.19(m,2H),3.94-3.85(m,2H),3.77-3.69(m,2H),3.56-3.48(m,6H),3.12-3.01( s, 6H), 2.87-2.64 (m, 2H), 2.37-2.18 (m, 10H), 1.33-1.22 (m, 6H), 1.15-0.91 (d, 2H), 0.53-0.36 (m, 8H), [M+H] ⁺ :m/z=833.8

Example 62:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.23-8.18 (s, 2H), 7.66-7.58 (m, 2H), 7.45-7.36 (m, 2H), 7.19-7.02 (m, 2H) , 4.88-4.72(d, 2H), 4.37-4.21(m, 2H), 4.04-3.89(m, 2H), 3.78-3.65(m, 2H), 3.58-3.46(m, 6H), 3.14-3.09( s,6H),2.83-2.64(m,2H),2.52-2.41(m,2H),2.30-2.16(m,10H),1.79-1.68(d,6H),1.43-1.24(m,20H), [M+H] ⁺ :m/z=918.3

Example 63:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.16-8.07(s,2H), 7.82-7.75(m,2H), 7.58-7.45(m,2H), 7.22-7.13(m,2H) ,4.86-4.64(d,2H),4.41-4.27(m,2H),3.98-3.79(m,2H),3.63-3.54(m,2H),3.41-3.30(m,6H),3.15-3.07( s,6H),2.83-2.71(m,2H),2.32-2.14(m,10H),1.65-1.56(m,6H),1.37-1.28(d,2H),0.99-0.81(d,12H), [M+H] ⁺ :m/z=852.2

Example 64:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.14-8.09 (s, 2H), 7.63-7.52 (m, 2H), 7.32-7.25 (m, 2H), 7.12-7.01 (m, 2H) ,4.63-4.55(d,2H),4.15-3.96(m,2H),3.83-3.72(m,2H),3.68-3.58(m,2H),3.43-3.32(m,6H),3.15-3.07( s,6H), 2.87-2.77(m,2H),2.36-2.17(m,10H),1.67-1.53(m,6H),1.33-1.22(d,3H),0.94-0.84(d,6H), [M+H] ⁺ :m/z=809.7

Example 65:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.10-8.02 (s, 2H), 7.64-7.52 (m, 2H), 7.45-7.34 (m, 2H), 7.27-7.18 (m, 2H) ,4.72-4.63(d,2H),4.35-4.19(m,2H),3.98-3.85(m,2H),3.75-3.66(m,2H),3.57-3.48(m,6H),3.23-3.14( s,6H),2.96-2.87(m,6H),2.45-2.34(m,10H),2.04-1.91(d,2H),1.34-1.28(d,3H),1.16-1.04(d,6H), [M+H] ⁺ :m/z=825.8

Example 66:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.18-8.11 (s, 2H), 7.78-7.66 (m, 2H), 7.56-7.39 (m, 2H), 7.15-7.06 (m, 2H) , 4.89-4.66(d,2H),4.22-4.13(m,2H),3.85-3.73(m,2H),3.57-3.43(m,2H),3.34-3.25(m,6H),3.16-3.05( s,6H),2.73-2.66(m,2H),2.45-2.23(m,10H),1.74-1.55(m,6H),1.35-1.28(s,9H),0.96-0.87(d,6H), [M+H] ⁺ :m/z=852.3

Example 67:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.20-8.11 (s, 2H), 7.84-7.75 (m, 2H), 7.54-7.43 (m, 2H), 7.18-7.02 (m, 2H) ,4.85-4.64(d,2H),4.24-4.13(m,2H),3.93-3.82(m,2H),3.74-3.66(m,2H),3.54-3.46(m,6H),3.25-3.17( s,6H),2.96-2.84(m,2H),2.46-2.34(m,10H),1.74-1.66(m,6H),1.46-1.36(m,1H),1.18-0.94(d,6H), 0.73-0.65(m,4H),[M+H] ⁺ :m/z=835.8

Example 68:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.18-8.06 (s, 2H), 7.66-7.55 (m, 2H), 7.45-7.36 (m, 2H), 7.16-7.07 (m, 2H) ,4.66-4.57(d,2H),4.19-3.96(m,2H),3.86-3.74(m,2H),3.65-3.57(m,2H),3.47-3.35(m,6H),3.15-3.10( s, 6H), 2.86-2.76 (m, 2H), 2.37-2.22 (m, 10H), 2.13-1.96 (m, 10H), 1.65-1.56 (m, 6H), 1.36-1.28 (d, 1H), 0.97-0.88(d,6H),[M+H] ⁺ :m/z=878.3

Example 69: Preparation of benzyloxycarbonyl (2S)-2-[(1,3-dioxoisoindol-2-yl)methyl]-4-fluoropyrrolidine-1-carboxylate

Take 100g (0.26mol) of benzyloxycarbonyl(2S)-2-[(1,3-dioxoisoindol-2-yl)methyl]-4-hydroxypyrrolidine-1-carboxylate in 1L three mouths Into the reaction flask, add dichloromethane (600mL), and under the protection of nitrogen, the system is cooled to -78℃, and DAST63.6g (0.41mol) is added dropwise at -78℃. After the addition is complete, stir at -78℃ After 3 hours, the system was warmed to room temperature and stirred for 16 hours. TLC detected that the reaction was complete. The reaction solution was poured into 800 mL of saturated sodium bicarbonate solution at 0°C for quenching, and extracted with dichloromethane (300 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:40) to obtain benzyloxycarbonyl (2S)-2-[(1,3-dioxoisoindol-2-yl) Methyl]-4-fluoropyrrolidine-1-carboxylate (22 g, 22%).

Example 70: Preparation of benzyloxycarbonyl (2S)-2-(aminomethyl)-4-fluoropyrrolidine-1-carboxylate

Take benzyloxycarbonyl (2S)-2-[(1,3-dioxoisoindol-2-yl)methyl]-4-fluoropyrrolidine-1-carboxylate 22g (57.6mmol) in 1L reaction Into the bottle, add 450 mL of ethanol, heat to 80°C, and add 5.7 g (0.14 mol, 85%) of hydrazine hydrate to it dropwise. Then it was stirred at 80°C for 12 hours. TLC detected that the reaction was complete. Cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain benzyloxycarbonyl (2S)-2-(aminomethyl)-4-hydroxypyrrolidine-1-carboxylate (13.5g, 93%), which was used directly for the next One step response.

Example 71: Preparation of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-nitro-anilino)methyl]-4-fluoropyrrolidine-1-carboxylate

Take 13.5g (53.6mmol) of benzyloxycarbonyl (2S)-2-(aminomethyl)-4-fluoropyrrolidine-1-carboxylate in a 500mL reaction flask, add 250mL of acetonitrile, and take 14.8g of potassium carbonate. 0.11 mol) was added to it, and 10.2 g (64.3 mmol) of 1,4-difluoro-2-nitrobenzene was added to it. Under the protection of nitrogen, the temperature was raised to 80°C and stirred for 1 hour. TLC detected that the reaction was complete, quenched by adding 200 mL of water, and extracted with ethyl acetate (200 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:40) to obtain benzyloxycarbonyl (2S)-2-[(4-fluoro-2-nitro-anilino)methyl] -4-fluoropyrrolidine-1-carboxylate (18.1 g, 86%).

Example 72: Preparation of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]-4-fluoropyrrolidine-1-carboxylate

Take 18.1 g (46.3 mmol) of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-nitro-anilino)methyl]-4-fluoropyrrolidine-1-carboxylate in a 500mL reaction flask Add 200 mL of acetic acid, and add 25 g of iron powder to it. Stir at room temperature for 16 hours. TLC detects that the reaction is complete. Filtered. The filter cake is rinsed with ethyl acetate. The filtrate is concentrated under reduced pressure. Add 300 mL of water and ethyl acetate (200 mL×3). ) Extraction, the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Obtained benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]-4-fluoropyrrolidine-1-carboxylate (15.9g, 95%), which was used directly for the next One step response.

Example 73: (2S)-Benzyl 4-fluoro-2-((5-fluoro-2-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)pyrrolidine-1 -Preparation of carboxylic acid esters

Take 15.9 g (44.0 mmol) of benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]-4-fluoropyrrolidine-1-carboxylate in a 500 mL reaction flask, Add 300mL of 4N hydrochloric acid, and add another 10g (0.132mol) of acetic alcohol into it. Heat to 100°C and stir for 6 hours. TLC detects that the reaction is complete. Cool down to 0°C. Add saturated sodium bicarbonate solution dropwise to adjust pH=8-9, filter, The filter cake was washed with methyl tert-butyl ether and dried to obtain (2S)-benzyl 4-fluoro-2-((5-fluoro-2-(hydroxymethyl)-1H-benzo[d]imidazole-1- (Yl)methyl)pyrrolidine-1-carboxylate (15.7g, 89%), directly used in the next reaction.

Example 74: (2S)-Benzyl 4-fluoro-2-((5-fluoro-2-formyl-1H-benzo[d]imidazol-1-yl)methyl)pyrrolidine-1-carboxylic acid Preparation of ester

Take (2S)-benzyl 4-fluoro-2-((5-fluoro-2-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)pyrrolidine-1-carboxylic acid Put 15.7g (39.2mmol) of ester in a 500mL reaction flask, add 300mL of dichloromethane, under the protection of nitrogen, cool to 0°C, and add 20g (47.0mmol) of Des Martin reagent to it, return to room temperature, and stir for 2 hours. TLC detects that the reaction is complete, the temperature is reduced to 0°C, washed with NaHCO ₃ /Na ₂ S ₂ O ₃ (1:1) saturated solution (200 mL*2), the aqueous phase is extracted with dichloromethane (200 mL*2), and the combined The organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:5) to obtain (2S)-benzyl 4-fluoro-2-((5-fluoro-2-formyl-1H-benzene) And [d]imidazol-1-yl)methyl)pyrrolidine-1-carboxylate (14.4 g, 92%).

Example 75: (5S,5'S)-dibenzyl 5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenz[d]imidazole]-1 ,1'-Diyl)bis(methylene))bis(3-fluoropyrrolidine-1-carboxylate)

Take (2S)-benzyl 4-fluoro-2-((5-fluoro-2-formyl-1H-benzo[d]imidazol-1-yl)methyl)pyrrolidine-1-carboxylate 14.4g (36.1mmol), benzyloxycarbonyl (2S)-2-[(4-fluoro-2-amino-anilino)methyl]-4-fluoropyrrolidine-1-carboxylate 15.6g (43.3mmol) in 250mL Into the reaction flask, add 150mL of N,N-dimethylformamide, add 27.4g (0.14mol) of sodium metabisulfite, under nitrogen protection, under microwave conditions, reflux and stir for 1 hour, TLC detection reaction is complete, return to room temperature, add 500mL of water, acetic acid Ethyl acetate (200 mL×3) was extracted, and the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:5) to obtain (5S,5'S)-dibenzyl 5,5'-((5,5'-difluoro-1H, 1'H-[2,2'-Dibenzyl[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-fluoropyrrolidine-1-carboxylate) (22.2 g, 83%).

Example 76: 5,5'-Difluoro-1,1'-bis((2S)-4-fluoropyrrolidin-2-yl)methyl)-1H,1'H-2,2'-biben Preparation of [d]imidazole

Take (5S,5'S)-dibenzyl 5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzyl[d]imidazole]-1,1' -Diyl) bis(methylene)) bis(3-fluoropyrrolidine-1-carboxylate) 22.2g (30.0mmol) in a 250mL reaction flask, add 100mL methanol, another 2g palladium charcoal added to it, hydrogen Under atmosphere, stirring at room temperature for 12 hours, TLC detected that the reaction was complete, filtered, and the filtrate was concentrated to obtain 5,5'-difluoro-1,1'-bis((2S)-4-fluoropyrrolidin-2-yl)methyl )-1H,1'H-2,2'-bibenzo[d]imidazole (13.6g, 95% crude product), directly used in the next reaction.

Example 77: Di-tert-butyl ((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2' -Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-fluoropyrrolidine-5,1-diyl))bis(3-methyl-1- Preparation of oxobutane-2,1-diyl)) dicarbamate

Take 2.2g (10.2mmol) of N-Boc-L valine in a 100mL reaction flask, add 20mL of N,N-dimethylformamide, and take another 2.1g (21.2mmol) of N-methylmorpholine, HATU4.8g (12.7mmol) was added to it, stirred at room temperature for 30 minutes, and then 5,5'-difluoro-1,1'-bis((2S)-4-fluoropyrrolidin-2-yl)methyl)-1H,1 'H-2,2'-Bibenzo[d]imidazole 2g (4.2mmol) was dissolved in 20mL of N,N-dimethylformamide, added dropwise to the above system, stirred at room temperature for 3 hours, TLC detected that the reaction was complete, add 200mL It was extracted with water and ethyl acetate (200 mL×3), and the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:3) to obtain di-tert-butyl ((2R, 2'R)-((5S,5'S)-5,5'-( (5,5'-Difluoro-1H,1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-fluoro Pyrrolidine-5,1-diyl))bis(3-methyl-1-oxobutane-2,1-diyl))dicarbamate (2.2 g, 60%).

Example 78: (2R,2'R)-1,1'-((5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2' -Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-fluoropyrrolidine-5,1-diyl))bis(2-amino-3-methyl Butan-1-one) preparation

Take di-tert-butyl ((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2'-diphenyl And [d]imidazole]-1,1'-diyl)bis(methylene))bis(3-fluoropyrrolidine-5,1-diyl))bis(3-methyl-1-oxobutane) Alkane-2,1-diyl)) dicarbamate 1.0g (1.15mmol) was added to a 100mL reaction flask, 20mL of hydrochloric acid ethanol solution was added, and the mixture was stirred at room temperature for 1 hour. TLC detected that the reaction was complete, and the solvent and excess were evaporated under reduced pressure. Hydrogen chloride, 0.79 g of colorless oily liquid was obtained, which was directly used in the next reaction.

Example 79: Di-tert-butyl ((2R,2'R)-((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H, 1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-fluoropyrrolidine-5,1-diyl)) Bis(3-methyl-1-oxobutane-2,1-diyl))bis(azadiyl))bis(1-oxopropane-2,1-diyl))bis(methylamino) Preparation of methyl formate)

Take 0.58g (2.88mmol) of N-Boc-N-methyl-L alanine in a 100mL reaction flask, add 20mL of N,N-dimethylformamide, and take another 0.61g (6.0mmol) of N-methylmorpholine ), HATU1.37g (3.6mmol) was added to it, stirred at room temperature for 30 minutes, and then (2R,2'R)-1,1'-((5S,5'S)-5,5'-((5,5' -Difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-fluoropyrrolidine-5, 1-Diyl)) bis(2-amino-3-methylbutan-1-one) 0.79g (crude product, 1.15mmol) dissolved in 20mL N,N-dimethylformamide, added dropwise to the above system, room temperature After stirring for 3 hours, TLC detected that the reaction was complete, 200 mL of water was added, and extraction was performed with ethyl acetate (200 mL×3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:3) to obtain di-tert-butyl ((2R, 2'R)-((2R, 2'R)-((5S, 5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]-1,1'-diyl) bis( Methyl))bis(3-fluoropyrrolidine-5,1-diyl))bis(3-methyl-1-oxobutane-2,1-diyl))bis(azadiyl)) Bis(1-oxopropane-2,1-diyl))bis(methylcarbamate) 0.57 g (48%).

Example 80: (2R,2'R)-N,N'-((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H, 1'H-[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-fluoropyrrolidine-5,1-diyl)) Preparation of bis(3-methyl-1-oxobutane-2,1-diyl))bis(2-(methylamino)propionamide)

Take the di-tert-butyl ((2R,2'R)-((2R,2'R)-((5S,5'S)-5,5'-((5,5'-difluoro-1H,1'H -[2,2'-Dibenzo[d]imidazole]-1,1'-diyl)bis(methylene))bis(3-fluoropyrrolidine-5,1-diyl))bis(3 -Methyl-1-oxobutane-2,1-diyl))bis(azadiyl))bis(1-oxopropane-2,1-diyl))bis(methylcarbamate) ) 0.57g (0.55mmol) in a 100mL reaction flask, add 20mL of hydrochloric acid ethanol solution, stir at room temperature for 1 hour, TLC detects that the reaction is complete, the solvent and excess hydrogen chloride are evaporated under reduced pressure, 100mL of water is added, saturated sodium bicarbonate is adjusted to pH=8 , Ethyl acetate extraction (100 mL*2), the combined organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified with a silica gel column (eluent: ethyl acetate: petroleum ether = 1:2) to obtain (2R,2'R)-N,N'-((2R,2'R)-((5S, 5'S)-5,5'-((5,5'-difluoro-1H,1'H-[2,2'-dibenzo[d]imidazole]-1,1'-diyl) bis( Methyl))bis(3-fluoropyrrolidine-5,1-diyl))bis(3-methyl-1-oxobutane-2,1-diyl))bis(2-(methylamino) Propionamide) 269 mg (58%).

¹ H-NMR(400MHz,d ₆ -DMSO): δ=8.11-8.05(s,2H),7.55-7.43(m,2H),7.35-7.25(m,2H),7.17-7.09(m,2H) ,4.65-4.55(d,2H),4.16-3.99(m,2H),3.84-3.75(m,2H),3.63-3.55(m,2H),3.45-3.35(m,4H),3.15-3.07( s,6H),2.73-2.64(m,2H),2.24-2.06(m,10H),1.35-1.26(m,6H),0.96-0.85(d,12H),(M+H) ⁺ :m/ z=841.8

With reference to the synthetic methods of Examples 69-80 and the first synthetic scheme, the following example compounds were synthesized:

Example 81:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.16-8.07(s,2H), 7.57-7.44(m,2H),7.34-7.26(m,2H),7.14-6.98(m,2H) ,4.74-4.63(d,2H),4.24-4.16(m,2H),3.91-3.82(m,2H),3.73-3.64(m,2H),3.55-3.40(m,4H),3.15-3.03( s,6H),2.86-2.64(m,2H),2.35-2.12(m,10H),1.33-1.26(m,6H),1.11-0.94(d,6H),(M+H) ⁺ :m/ z=785.7

Example 82:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.22-8.13 (s, 2H), 7.69-7.58 (m, 2H), 7.31-7.22 (m, 2H), 7.14-7.07 (m, 2H) ,4.66-4.58(d,2H),4.02-3.94(m,2H),3.84-3.76(m,2H),3.63-3.52(m,2H),3.44-3.33(m,4H),3.15-3.03( s,6H),2.73-2.66(m,2H),2.23-2.09(m,10H),1.33-1.23(m,6H),0.96-0.85(d,6H),(M+H) ⁺ :m/ z=813.8

Example 83:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.10-8.02 (s, 2H), 7.57-7.43 (m, 2H), 7.31-7.23 (m, 2H), 7.16-7.08 (m, 2H) ,4.64-4.53(d,2H),4.05-3.93(m,2H),3.82-3.73(m,2H),3.64-3.52(m,2H),3.42-3.31(m,4H),3.17-3.08( s,6H),2.75-2.64(m,2H),2.22-2.08(m,10H),1.67-1.55(m,6H),1.34-1.26(m,6H),1.03-0.94(d,6H), [M+H] ⁺ :m/z=870.2

Example 84:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.21-8.13 (s, 2H), 7.64-7.46 (m, 2H), 7.30-7.23 (m, 2H), 7.14-7.06 (m, 2H) ,4.65-4.53(d,2H),4.06-3.93(m,2H),3.84-3.76(m,2H),3.64-3.57(m,2H),3.43-3.36(m,4H),3.14-3.06( s,6H),2.74-2.65(m,2H),2.14-2.07(m,10H),1.46-1.38(m,6H),1.06-0.93(d,24H),(M+H) ⁺ :m/ z=870.6

Example 85:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.18-8.06 (s, 2H), 7.62-7.54 (m, 2H), 7.45-7.38 (m, 2H), 7.14-7.07 (m, 2H) , 4.86-4.77(d, 2H), 4.53-4.44(m, 2H), 4.19-4.13(m, 2H), 3.97-3.82(m, 6H), 3.70-3.62(m, 2H), 3.45-3.38( m,4H),3.14-3.08(s,6H),2.84-2.75(m,2H),2.42-2.35(m,10H),1.57-1.47(m,6H),(M+H) ⁺ :m/ z=817.7

Example 86:

¹ H-NMR(400MHz,d ₆ -DMSO): δ=8.12-8.04(s,2H), 7.73-7.65(m,2H),7.45-7.33(m,2H),7.21-7.13(m,2H) ,4.99-4.84(d,2H),4.55-4.48(m,2H),4.11-4.03(m,2H),3.83-3.72(m,6H),3.61-3.54(m,2H),3.42-3.37( m,4H),3.15-3.08(s,6H),2.84-2.78(m,2H),2.47-2.39(m,10H),1.54-1.48(m,6H),1.21-1.14(m,6H)[ M+H] ⁺ :m/z＝810.2

Example 87:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.12-8.06 (s, 2H), 7.66-7.49 (m, 2H), 7.36-7.24 (m, 2H), 7.16-7.08 (m, 2H) ,4.62-4.54(d,2H),4.14-3.99(m,2H),3.90-3.83(m,2H),3.76-3.63(m,2H),3.54-3.45(m,4H),3.32-3.19( s,6H),2.64-2.56(m,2H),2.33-2.17(m,10H),1.72-1.64(m,4H),1.34-1.21(m,6H),0.95-0.81(d,12H), [M+H] ⁺ :m/z=870.3

Example 88:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.18-8.07 (s, 2H), 7.56-7.45 (m, 2H), 7.37-7.27 (m, 2H), 7.18-6.99 (m, 2H) ,4.76-4.58(d,2H),4.24-4.13(m,2H),3.92-3.80(m,2H),3.72-3.61(m,2H),3.56-3.41(m,4H),3.15-3.03( s, 6H), 2.86-2.68 (m, 2H), 2.44-2.25 (m, 10H), 1.33-1.18 (m, 6H), 1.05-0.91 (d, 2H), 0.58-0.31 (m, 8H), [M+H] ⁺ :m/z=837.8

Example 89:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.19-8.09 (s, 2H), 7.56-7.42 (m, 2H), 7.33-7.23 (m, 2H), 7.16-6.98 (m, 2H) ,4.73-4.52(d,2H),4.26-4.18(m,2H),3.93-3.76(m,2H),3.61-3.54(m,2H),3.45-3.38(m,4H),3.13-3.05( s, 6H), 2.82-2.63 (m, 2H), 2.55-2.42 (m, 2H), 2.35-2.13 (m, 10H), 1.75-1.67 (d, 6H), 1.43-1.11 (m, 20H), [M+H] ⁺ :m/z=922.4

Example 90:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.14-8.02 (s, 2H), 7.73-7.61 (m, 2H), 7.54-7.35 (m, 2H), 7.22-7.13 (m, 2H) ,4.76-4.58(d,2H),4.33-4.15(m,2H),3.86-3.72(m,2H),3.64-3.56(m,2H),3.43-3.36(m,4H),3.16-3.04( s,6H),2.84-2.77(m,2H),2.35-2.15(m,10H),1.67-1.55(m,6H),1.37-1.25(d,2H),0.98-0.74(d,12H), [M+H] ⁺ :m/z=820.2

Example 91:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.11-8.02(s,2H), 7.64-7.54(m,2H),7.45-7.36(m,2H),7.14-7.06(m,2H) ,4.63-4.52(d,2H),4.15-4.06(m,2H),3.92-3.84(m,2H),3.69-3.57(m,2H),3.44-3.32(m,4H),3.17-3.08( s, 6H), 2.87-2.76 (m, 2H), 2.37-2.18 (m, 10H), 1.68-1.58 (m, 6H), 1.35-1.26 (d, 3H), 0.96-0.87 (d, 6H), [M+H] ⁺ :m/z=813.8

Example 92:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.16-8.04(s, 2H), 7.66-7.58(m, 2H), 7.46-7.37(m, 2H), 7.28-7.09(m, 2H) ,4.73-4.64(d,2H),4.38-4.17(m,2H),3.97-3.88(m,2H),3.78-3.66(m,2H),3.52-3.43(m,4H),3.26-3.17( s,6H),2.92-2.84(m,6H),2.42-2.33(m,10H),2.03-1.91(d,2H),1.31-1.22(d,3H),1.02-0.88(d,6H), [M+H] ⁺ :m/z=829.8

Example 93:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.23-8.14 (s, 2H), 7.77-7.66 (m, 2H), 7.56-7.35 (m, 2H), 7.16-7.08 (m, 2H) ,4.88-4.65(d,2H),4.22-4.14(m,2H),3.87-3.77(m,2H),3.53-3.44(m,2H),3.33-3.22(m,4H),3.17-3.08( s,6H),2.72-2.62(m,2H),2.48-2.27(m,10H),1.77-1.57(m,6H),1.35-1.26(s,9H),0.98-0.89(d,6H), [M+H] ⁺ :m/z=855.7

Example 94:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.21-8.13 (s, 2H), 7.83-7.72 (m, 2H), 7.57-7.36 (m, 2H), 7.25-7.16 (m, 2H) ,4.85-4.68(d,2H),4.28-4.15(m,2H),3.95-3.84(m,2H),3.74-3.65(m,2H),3.56-3.44(m,4H),3.24-3.14( s,6H),2.93-2.83(m,2H),2.43-2.34(m,10H),1.74-1.63(m,6H),1.42-1.35(m,1H),1.06-0.83(d,6H), 0.65-0.52(m,4H),[M+H] ⁺ :m/z=840.1

Example 95:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.16-8.05 (s, 2H), 7.66-7.57 (m, 2H), 7.43-7.34 (m, 2H), 7.17-7.08 (m, 2H) ,4.68-4.56(d,2H),4.13-3.97(m,2H),3.87-3.75(m,2H),3.65-3.53(m,2H),3.43-3.35(m,4H),3.16-3.07( s, 6H), 2.85-2.77 (m, 2H), 2.37-2.13 (m, 10H), 2.04-1.85 (m, 10H), 1.65-1.56 (m, 6H), 1.38-1.29 (d, 1H), 0.96-0.84(d,6H),[M+H] ⁺ :m/z=882.4

Example 96:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.18-8.05 (s, 2H), 7.53-7.42 (m, 2H), 7.35-7.26 (m, 2H), 7.16-7.07 (m, 2H) ,4.67-4.57(d,2H),4.03-3.92(m,2H),3.83-3.72(m,2H),3.67-3.55(m,3H),3.45-3.37(m,5H),3.18-3.09( s,6H),2.77-2.63(m,2H),2.22-2.03(m,10H),1.37-1.26(m,6H),0.94-0.86(d,12H),(M+H) ⁺ :m/ z=822.1

Example 97:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.11-8.04 (s, 2H), 7.52-7.43 (m, 2H), 7.34-7.25 (m, 2H), 7.15-7.06 (m, 2H) ,4.67-4.58(d,2H),4.05-3.97(m,2H),3.85-3.74(m,2H), 3.67-3.56(m,2H),3.45-3.37(m,5H),3.15-3.05( s,6H),2.74-2.65(m,2H),2.26-2.04(m,10H),1.33-1.22(m,6H),0.92-0.84(d,12H),(M+H) ⁺ :m/ z=823.8

Example 98:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.17-8.04 (s, 2H), 7.54-7.46 (m, 2H), 7.37-7.24 (m, 2H), 7.13-7.05 (m, 2H) ,4.65-4.54(d,2H),4.03-3.95(m,2H),3.85-3.75(m,2H),3.64-3.56(m,2H),3.47-3.37(m,5H),3.14-3.05( s,6H),2.73-2.63(m,2H),2.25-2.03(m,10H),1.37-1.26(m,6H),0.96-0.85(d,12H),(M+H) ⁺ :m/ z=839.7

Example 99:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.08-8.01 (s, 2H), 7.52-7.43 (m, 2H), 7.34-7.22 (m, 2H), 7.14-7.08 (m, 2H) ,4.64-4.56(d,2H),4.04-3.92(m,2H),3.82-3.74(m,4H),3.66-3.57(m,2H),3.47-3.33(m,6H),3.13-3.05( s,6H),2.75-2.62(m,2H),2.23-2.08(m,10H),1.38-1.22(m,6H),0.92-0.80(d,12H),(M+H) ⁺ :m/ z=820.3

Example 100:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.09-8.00 (s, 2H), 7.52-7.41 (m, 2H), 7.31-7.22 (m, 2H), 7.14-7.05 (m, 2H) ,4.65-4.52(d,2H),4.03-3.94(m,2H),3.85-3.73(m,2H),3.64-3.55(m,2H),3.46-3.33(m,6H),3.13-3.05( s,6H),2.75-2.65(m,6H),2.26-2.03(m,10H),1.32-1.21(m,6H),0.92-0.82(d,12H),(M+H) ⁺ :m/ z=834.6

Example 101:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.17-8.02 (s, 2H), 7.52-7.43 (m, 2H), 7.31-7.23 (m, 2H), 7.14-7.01 (m, 6H) ,4.66-4.55(d,2H),4.03-3.93(m,6H),3.82-3.72(m,2H),3.65-3.56(m,2H),3.47-3.36(m,6H),3.14-3.07( s,6H),2.77-2.64(m,2H),2.22-2.05(m,10H),1.36-1.23(m,6H),0.92-0.81(d,12H),(M+H) ⁺ :m/ z=910.4

Example 102:

1H-NMR (400MHz, d6-DMSO): δ=8.14-8.07(s, 2H), 7.54-7.44(m, 2H), 7.32-7.25(m, 2H), 7.13-7.02(m, 2H), 4.64 -4.56(d,2H),4.06-3.93(m,2H),3.83-3.72(m,2H),3.66-3.58(m,2H),3.48-3.35(m,4H),3.14-3.07(s, 6H),2.77-2.69(m,6H),2.29-2.07(m,10H),1.35-1.27(m,6H),0.95-0.86(d,12H),[M+H]+:m/z= 832.6

Example 103:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.11-8.02 (s, 2H), 7.54-7.45 (m, 2H), 7.33-7.26 (m, 2H), 7.13-7.02 (m, 2H) ,4.63-4.55(d,2H),4.07-3.98(m,2H),3.86-3.78(m,2H),3.67-3.53(m,4H),3.42-3.34(m,6H),3.17-3.05( s,6H),2.74-2.66(m,6H),2.22-2.04(m,10H),1.34-1.24(m,6H),0.95-0.86(d,12H),(M+H) ⁺ :m/ z=848.3

Embodiment 104:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.13-8.04 (s, 2H), 7.55-7.42 (m, 2H), 7.33-7.25 (m, 2H), 7.13-7.03 (m, 2H) ,4.66-4.59(d,2H),4.04-3.95(m,2H),3.83-3.73(m,2H),3.62-3.55(m,4H),3.46-3.38(m,6H),3.15-3.04( s,6H),2.73-2.63(m,6H),2.27-2.08(m,10H),1.39-1.25(m,6H),0.94-0.84(d,12H),(M+H) ⁺ :m/ z=862.3

Example 105:

¹ H-NMR (400MHz, d ₆ -DMSO): δ = 8.19-8.13 (s, 2H), 7.59-7.46 (m, 2H), 7.35-7.23 (m, 2H), 7.14-7.05 (m, 2H) ,4.62-4.53(d,2H),4.02-3.93(m,3H),3.84-3.71(m,4H), 3.60-3.55(m,6H),3.41-3.33(m,6H),3.13-3.01( s,6H),2.84-2.63(m,6H),2.23-2.07(m,10H),1.32-1.22(m,6H),0.92-0.85(d,12H),(M+H) ⁺ :m/ z=888.6

Example 106:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.15-8.03 (s, 2H), 7.53-7.45 (m, 2H), 7.36-7.24 (m, 2H), 7.16-7.07 (m, 6H) , 4.67-4.57(d,2H),4.05-3.96(m,6H),3.86-3.73(m,2H),3.66-3.53(m,2H),3.44-3.35(m,6H),3.15-3.04( s,6H),2.76-2.67(m,2H),2.27-2.08(m,10H),1.33-1.22(m,6H),0.96-0.82(d,12H),(M+H) ⁺ :m/ z=882.5

Example 107: Test of the binding affinity of the compound to XIAP, cIAP1, and cIAP2

The Bir3 structure region (10 nM) and Smac polypeptide (10 nM) were incubated in the test buffer (50 mM Tris, 120 mM Nacl, 0.1% BSA, 1 mM DTT, 0.05% TritonX100) in the presence of the test compound for 1 h at room temperature. The mixture is transferred to a streptavidin-coated plate, and then incubated at room temperature for 1 hour to allow the biotin-linked peptide and the Bir3 structure region to bind to the plate. After several washes, Eu-labeled anti-GST antibody (Perkin Elmer DELFIA Eu-N1-anti-GST, #AD0250) was added to each well (diluted 1:5000 with Perkin Elmer DELFIA test buffer 2013-01 ), then incubate at room temperature for 1h. After washing 3 times in DELFIA wash buffer (Perkin Elmer DELFIA Wash2013-05), add enhancement solution (Perkin Elmer enhancement solution 2013-02), incubate for 10 minutes, and then detect the intensity of time-resolved fluorescence europium, calculated with GraphPad Prizm5.0 software compound inhibition IC ₅₀ values. The specific test results are shown in Table 1 and Table 2 below. Note: (A:<10nM B:10-100nM C:>100nM)

Table 1 Test results of the binding affinity of the compound of the present invention to XIAP, cIAP1, and cIAP2

Table 2 Binding affinity test results of the compound of the present invention and the reference drug with XIAP, cIAP1, and cIAP2

The experimental results found that the compounds of the present invention have good binding affinity to XIAP, cIAP1, and cIAP2 proteins, and have good IAP inhibitory activity. According to the comparison results in Table 1, it can be further seen that some of the compounds of the present invention are effective against XIAP, cIAP1, and cIAP2 proteins. The binding affinity is significantly better than the control drug.

Example 108: Cell growth inhibition test in MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines

First collect the logarithmic growth phase cells. Count, resuspend the cells in complete medium, adjust the cell concentration to an appropriate concentration (determined according to the cell density optimization test results), inoculate a 96-well plate, and add 100 μl of cell suspension to each well. Place the cells in a 37°C, 100% relative humidity, 5% CO ₂ incubator and incubate for 24 hours. The compound of the test example was diluted with culture medium to the corresponding concentration, and 25 μl/well was added to the cells. The final concentration of the compound starts from 100 μM, and is diluted 4-fold, with a total of 10 concentration points. Place the cells in a 37°C, 100% relative humidity, 5% CO ₂ incubator for 72 hours. Directly add 10μl of CCK-8 to the cell culture medium and incubate in a 37°C incubator for 2-4 hours. After shaking gently, the absorbance was measured on the SpectraMax M5 Microplate Reader, and the inhibition rate was calculated. The specific test results are shown in Table 3 and Table 4 below.

Note: (A: <10nM B: 10-100nM C: 100-1000nM D:>1000nM)

Table 3 Inhibition results of the compounds of the present invention on MDA-MB-231 and PC-3 cell lines

Table 4 Inhibition results of the compounds of the present invention and control drugs on MDA-MB-231 and PC-3 cell lines

The compound of the present invention has a good inhibitory effect on cell growth in MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines. It can be seen from Table 4 that some compounds have significantly better inhibitory effects on MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines than the control drugs.

In the foregoing, the embodiments of the present invention have been described. However, the present invention is not limited to the above-mentioned embodiment. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. A compound represented by formula I and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or the like Pharmaceutically acceptable salt:

   

   Wherein, R ₁ and R ₂ are independently selected from;

   

   R ₃ , R ₄ and R ₅ are each independently selected from H or optionally substituted by 1, 2 or 3 R: (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one, two or more Heteroatom (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, C _3-12 cycloalkyl group, 3-12 membered heterocyclic group, C _6-20 aryl group or 5-14 membered heteroaryl group;

   Ring A and Ring B are each independently selected from: C _6-20 aryl or 5-14 membered heteroaryl;

   R ₆ and R ₇ are each independently selected from halogen, hydroxyl, or selected from optionally substituted by 1, 2 or 3 R: (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one, two or more A heteroatom (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, a C _3-12 cycloalkyl group, a 3-12 membered heterocyclic group, a C _6-20 aryl group or a 5-14 membered heteroaryl group;

   m and n are independently selected from: 0, 1, 2 or 3;

   R is selected from halogen, CN, OH, SH, NH ₂ , COOH, or selected from optionally substituted by 1, 2 or 3 R': (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one or two _{(C 1} -C ₁₂ ) aliphatic hydrocarbon group with one or more heteroatoms _{, C 3-12} cycloalkyl, 3-12 membered heterocyclic group, C _6-20 aryl group or 5-14 membered heteroaryl group;

   R'is selected from halogen, CN, OH, SH, NH ₂ , COOH; P ₁ and P ₂ are independently selected from halogen, OH, CN, NH ₂ , COOH, (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally _{A (C 1} -C ₁₂ ) aliphatic hydrocarbon group containing one, two or more heteroatoms.

   W is selected from a single bond, -O-, -S-, -NH- or optionally substituted by 1, 2 or 3 R: (C ₁ -C ₁₂ ) aliphatic hydrocarbon group, optionally including one, two or (C ₁ -C ₁₂ ) aliphatic hydrocarbon groups with _{more heteroatoms, C 3-12} cycloalkyl groups, 3-12 membered heterocyclic groups, C _6-20 aryl groups or 5-14 membered heteroaryl groups.
2. A compound of formula I according to claim 1 and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, and metabolites , Ester, prodrug or a pharmaceutically acceptable salt thereof, characterized in that in the "(C ₁ -C ₁₂ )aliphatic hydrocarbon group optionally containing one, two or more heteroatoms", the The heteroatom can be selected from sulfur, nitrogen, oxygen, phosphorus and silicon. Optionally, the heteroatom is inserted into the aliphatic hydrocarbon group, optionally a CC bond and a CH bond; for example, it can be selected from (C ₁ -C ₁₂ ) aliphatic hydrocarbon group oxy , (C ₁ -C ₁₂ ) aliphatic hydrocarbyl mercapto group, (C ₁ -C ₆ ) aliphatic hydrocarbyl oxy group, (C ₁ -C ₆ ) aliphatic hydrocarbyl mercapto group, (C ₁ -C ₆ ) aliphatic hydrocarbyl oxy group (C _1- C ₆ ) aliphatic hydrocarbon group, (C ₁ -C ₆ ) aliphatic hydrocarbon group mercapto group (C ₁ -C ₆ ) aliphatic hydrocarbon group, N-(C ₁ -C ₃ ) aliphatic hydrocarbon group amine group (C ₁ -C ₆ ) aliphatic hydrocarbon group, N , N-di-(C ₁ -C ₃ ) aliphatic hydrocarbyl amine group (C ₁ -C ₆ ) aliphatic hydrocarbyl; the "halogen" is selected from F, Cl, Br, and I.
3. A compound of formula I according to claim 1 and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, and metabolites , Ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that said R ₃ , R ₄ and R ₅ can be independently selected from the following groups: methyl, ethyl, n-propyl, isopropyl Base, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1 -Butenyl, 1-ethylvinyl, 1-methyl-2-propenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 2-methyl-2 -Propenyl, 1-pentenyl, 1-hexenyl, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 1-methyl-2-propynyl, 3- Butynyl, 1-pentynyl, 1-hexynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,

   
4. A compound of formula I according to claim 1 and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, and metabolites , Ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that the structure of formula I is further selected from the following formula II, formula III, formula IV, and formula V:

   

   In the formula II, formula III, formula IV, and formula V, R ₁ , R ₂ , R ₆ , R ₇ , P ₁ , P ₂ , W, n, m, ring A, and ring B are as defined in formula I.
5. A compound of formula I according to claim 1 and its racemates, stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, and metabolites , Ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that the compound represented by formula I is selected from:

   

   

   

   

   

   

   

   
6. A compound of formula I according to any one of claims 1 to 5, and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, and polymorphs thereof The preparation method of the form, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that, the preparation method comprises the following steps:

   

   

   _{Wherein, R 1, R 2, R} 3, R 4, R 5, R 6, R 7, P 1, P 2, W as defined for formula I; R ₃ 'is selected from the group R ₃ defined range, And independently selected from groups different _{from R 3 ; PG 1} and PG ₂ respectively represent different amino protecting groups, which can be selected from tert-butoxycarbonyl (N-boc), benzyloxycarbonyl protecting groups (N-cbz ), fluorenylmethoxycarbonyl protecting group (N-Fmoc), preferably, PG _{1 is} selected from tert-butoxycarbonyl, and PG _{2 is} selected from benzyloxycarbonyl protecting group.
7. A compound of formula I according to any one of claims 1 to 5, and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, and polymorphs thereof The preparation method of the form, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, characterized in that, the preparation method comprises the following steps:

   

   

   _{Wherein, R 1, R 2, R} 3, R 4, R 5, R 6, R 7, P 1, P 2, W as defined for formula I; R ₃ 'is selected from the group R ₃ defined range, And independently selected from groups different _{from R 3 ; PG 1} and PG ₂ respectively represent different amino protecting groups, selected from tert-butoxycarbonyl (N-boc) and benzyloxycarbonyl protecting groups (N-cbz) , Fluorenylmethoxycarbonyl protecting group (N-Fmoc), preferably, PG _{1 is} selected from tert-butoxycarbonyl, and PG _{2 is} selected from benzyloxycarbonyl protecting group.
8. A pharmaceutical composition comprising a compound represented by formula I according to any one of claims 1 to 5, and its racemates, stereoisomers, tautomers, isotope markers, and nitrogen oxides , Solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof.
9. According to any one of claims 1-5, a compound of formula I and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, Use of metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof in the preparation of inhibitors of apoptosis proteins.
10. According to any one of claims 1-5, a compound of formula I and its racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, Use of metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof in drugs for treating diseases or disorders caused by IAP disorders, the diseases or disorders selected from a variety of benign tumors or malignant tumors (cancer), benign Proliferative diseases (e.g., psoriasis, benign prostatic hypertrophy, and restenosis), or autoimmune diseases (e.g., autoimmune proliferative glomerulonephritis, lymphoproliferative autoimmune response). Cancers that can be treated with IAP antagonists include but are not limited to one or more of the following cancers: lung adenocarcinoma, pancreatic cancer, colon cancer, ovarian cancer, breast cancer, mesothelioma, peripheral neuroma, bladder cancer , Glioblastoma, melanoma, adrenal cortical carcinoma, AIDS-related lymphoma, anal cancer, bladder cancer, meningioma, glioma, astrocytoma, breast cancer, cervical cancer, Chronic myelodysplastic disorders (e.g., chronic lymphocytic leukemia, chronic myelogenous leukemia), colon cancer, endocrine adenocarcinoma, endometrial cancer, ependymoma, esophageal cancer, Ewing’s sarcoma, extracranial germ cell tumor, Extragonadal germ cell tumor, extrahepatic cholangiocarcinoma, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gestational trophoblastoma, hairy cell leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hypopharyngeal carcinoma, Intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, laryngeal cancer, leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, lip cancer, oral cancer, liver cancer, male breast cancer, malignant mesothelioma , Medulloblastoma, melanoma, Merkel cell carcinoma, metastatic squamous neck cancer, multiple myeloma and other plasma cell neoplasms, mycosis fungoides and Sezary syndrome (Sezary syndrome), myelodysplastic syndrome, nasopharyngeal carcinoma, neuroblastoma, non-small cell lung cancer, small cell lung cancer, oropharyngeal cancer, bone cancer (including osteosarcoma and bone malignant fibrous histiocytoma), ovarian epithelial cancer , Ovarian germ cell tumors, ovarian low malignant potential tumors, pancreatic cancer, paranasal sinus cancer, parathyroid cancer, penile cancer, pheochromocytoma, pituitary tumor, prostate cancer, rectal cancer , Renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, small intestine cancer, soft tissue sarcoma, supratentorial primary neuroectodermal tumor, pineoblastoma, testicular cancer, thymoma, thymic cancer , Thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer, as well as Wilm's tumor and other childhood kidney tumors.