CN112521372A - Apoptosis protein inhibitor and preparation method and application thereof - Google Patents

Abstract

The invention relates to a compound shown in formula I, and racemate, stereoisomer, tautomer, isotopic marker, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the compound, a preparation method thereof, and medical application thereof, wherein the structure of the formula I is as follows:

Description

Apoptosis protein inhibitor and preparation method and application thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to an apoptosis protein inhibitor, and a preparation method and application thereof.

Background

Apoptosis (apoptosis) refers to the autonomous, ordered death of cells under the control of genes to maintain homeostasis. It plays an important role in the evolution of organisms, the stabilization of the internal environment and the development of a number of systems. Apoptosis signaling is divided into intrinsic (mediated by death receptor-ligand interactions) and extrinsic (mediated by cellular stress and mitochondrial permeability). Both pathways eventually converge with a Caspase (Caspase). Once the apoptotic signal is activated, caspases cleave a number of substrates associated with cell death, causing cell death. Inhibitor of apoptosis inhibitor proteins (IAPs) are a highly conserved family of endogenous anti-apoptotic factors that inhibit apoptosis primarily through inhibition of Caspase activity and involvement in the regulation of nuclear factor NF- κ B. SMAC mimetics (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, and when administered to an animal suffering from a proliferative disease, antagonize IAP, resulting in increased apoptosis in abnormally proliferating cells. There is a need to develop new IAP antagonists with better activity, selectivity and safety.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides a compound shown as formula I, and racemate, stereoisomer, tautomer, isotopic marker, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof:

wherein R is₁And R₂Each is independently selected from;

R₃、R₄and R₅Each independently selected from H or optionally substituted with 1,2 or 3R: (C)₁-C₁₂) Fat and oil(C) an aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, C_6-20Aryl or 5-14 membered heteroaryl;

ring a and ring B are each independently selected from: c_6-20Aryl or 5-14 membered heteroaryl;

R₆and R₇Each independently selected from halogen, hydroxy or from optionally substituted by 1,2 or 3R: (C)₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, C_6-20Aryl or 5-14 membered heteroaryl;

m and n are each independently selected from: 0.1, 2 or 3;

r is selected from halogen, CN, OH, SH, NH₂COOH, or selected from optionally substituted with 1,2 or 3R': (C)₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, C_6-20Aryl or 5-14 membered heteroaryl;

r' is selected from halogen, CN, OH, SH, NH₂、COOH；P₁And P₂Each independently selected from halogen, OH, CN, NH₂、COOH、(C₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) An aliphatic hydrocarbon group.

W is selected from a single bond, -O-, -S-, -NH-or optionally substituted with 1,2 or 3R: (C)₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, C_6-20Aryl or 5-14 membered heteroaryl.

In accordance with an embodiment of the present invention,

said "optionally comprising one, two or more heteroatoms of (C)₁-C₁₂) Aliphatic hydrocarbon group, the heteroThe atoms may be selected from sulphur, nitrogen, oxygen, phosphorus and silicon, optionally heteroatoms inserted into the aliphatic hydrocarbon groups, optionally C-C bonds and C-H bonds. (ii) a For example, may be selected from (C)₁-C₁₂) Aliphatic hydrocarbyloxy, (C)₁-C₁₂) Aliphatic hydrocarbyl mercapto group, (C)₁-C₆) Aliphatic hydrocarbyloxy group (C)₁-C₆) Aliphatic hydrocarbyl mercapto group, (C)₁-C₆) Aliphatic hydrocarbyloxy (C)₁-C₆) Aliphatic hydrocarbon group, (C)₁-C₆) Aliphatic hydrocarbyl mercapto group (C)₁-C₆) Aliphatic hydrocarbon group, N- (C)₁-C₃) Aliphatic hydrocarbyl amino group (C)₁-C₆) Aliphatic hydrocarbon group, N-di- (C)₁-C₃) Aliphatic hydrocarbyl amino group (C)₁-C₆) An aliphatic hydrocarbon group;

said (C)₁-C₁₂) The aliphatic hydrocarbon group may be selected from (C)₁-C₁₂) Alkyl, (C)₂-C₁₂) Alkenyl, (C)₂-C₁₂) Alkynyl groups, in some embodiments, can be selected from (C)₁-C₆) Alkyl, (C)₂-C₆) Alkenyl, (C)₂-C₆) An alkynyl group;

according to embodiments of the invention, in some embodiments, the R is₃、R₄And R₅Can be selected from the following groups independently: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, ethenyl, 1-propenyl, 2-propenyl, 1-methylethenyl, 1-butenyl, 1-ethylethenyl, 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, tert-butyl,

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

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

in the formulas II, III, IV and V, R₁、R₂、R₆、R₇、P₁、P₂W, n, m, ring A, ring B are as defined above for formula I.

According to the invention, in the above formulae I-V, R₁And R₂Each is independently selected from;

R₃、R₄and R₅Each independently selected from H or the following optionally substituted with 1,2 or 3R: (C)₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl;

ring a and ring B are each independently selected from: c_6-20Aryl or 5-14 membered heteroaryl;

R₆and R₇Each independently selected from halogen or hydroxy;

m and n are each independently selected from: 0.1, 2 or 3;

r is selected from halogen, CN, OH, SH, NH₂COOH, or selected from optionally substituted with 1,2 or 3R': (C)₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, C_6-20Aryl or 5-14 membered heteroaryl;

r' is selected from halogen, CN, OH, SH, NH₂、COOH；

P₁And P₂Each independently selected from halogen, OH, NH₂、(C₁-C₁₂) An aliphatic hydrocarbon group.

W is selected from the group consisting of a single bond, -O-, -S-, -NH-.

Illustrative, non-limiting specific examples of the compounds of the present invention are shown below:

the invention also provides a preparation method of the compound shown in the formula I (including the compound shown in the formula II-formula V) and racemate, stereoisomer, tautomer, isotopic marker, nitrogen oxide, solvate, polymorphic substance, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, but not limited to the method described below. All starting materials are prepared or purchased directly according to the general rules of the target molecule and by protocols in these routes, methods well known to those of ordinary skill in the art of organic chemistry. The compounds of the invention can be synthesized by combining the methods described below with synthetic methods known in the art of synthetic organic chemistry or variations thereon as recognized by those skilled in the art.

In a first embodiment, the preparation of the compounds of the invention comprises the following steps:

in a second embodiment, the preparation of the compounds of the invention comprises the following steps:

in the scheme, M-9 can be obtained by adopting the preparation step (M-9 is prepared by taking M-1 as a starting material) in the first scheme.

According to an embodiment of the present invention, in both of the schemes,

R₁、R₂、R₃、R₄、R₅、R₆、R₇、P₁、P₂w is as defined for formula I;

R₃' selected from R₃Groups within the definition and independently selected from R₃Different groups;

PG₁、PG₂each represents a different amino protecting group selected from the group consisting of t-butyloxycarbonyl (N-boc), benzyloxycarbonyl protecting group (N-cbz), fluorenylmethoxycarbonyl protecting group (N-Fmoc), preferably, PG₁Selected from t-butyloxycarbonyl, PG₂Selected from benzyloxycarbonyl protecting groups.

According to an embodiment of the present invention, in both preparation schemes, M-9 can be prepared by using M-1 as a starting material, and the preparation of M-9 comprises the following steps:

(a1) with N-PG₁Protected prolinol M-1 is taken as a starting material and reacts with phthalimide under the action of triphenylphosphine, diisopropyl azodicarboxylate and the like to generate M-2;

preferably, in said step, the reaction temperature may be selected from 0 ℃ to 25 ℃, and the reaction solvent may be selected from inert aprotic solvents, for example selected from tetrahydrofuran, toluene, dichloromethane and the like;

(a2) performing hydrazinolysis on the M-2 under the condition of hydrazine hydrate to obtain M-3;

preferably, in the step, the reaction temperature may be selected from 25 ℃ to 80 ℃, and the reaction solvent may be selected from methanol, ethanol, tetrahydrofuran, and the like;

(a3) m-3 reacts with M-4 under the alkaline condition to generate M-5;

preferably, in the step, the reaction temperature may be selected from 70 ℃ to 90 ℃, the reaction solvent may be selected from acetonitrile, DMF, tetrahydrofuran, and the like, and the basic condition may be selected from carbonates of alkali metals or alkaline earth metals, for example, from potassium carbonate, sodium carbonate, and the like;

(a4) reducing the M-5 by iron powder to obtain M-6;

preferably, in said step, the reaction temperature may be selected from room temperature, for example from 25 ℃, and the reaction solvent may be selected from acetic acid;

(a5) m-6 and M-7 are subjected to ring closing in an acid system to obtain M-8;

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

(a6) oxidizing the hydroxyl of M-8 to obtain an aldehyde compound M-9;

preferably, in the step, the oxidant used is dessimutant reagent, the reaction temperature can be selected from 0 ℃ to 25 ℃, and the reaction solvent can be selected from inert aprotic solvents, such as tetrahydrofuran, dichloromethane, toluene and the like;

according to an embodiment of the invention, in the first variant, the process for preparing formula I from M-9 comprises the following steps:

(b1) closing the ring of M-9 and M-10 under the condition of a catalyst to obtain M-11;

preferably, in said step, the catalyst may be selected from Oxone, sodium metabisulfite, etc., the reaction temperature may be selected from 25 ℃ to 140 ℃, and the reaction solvent may be selected from inert high boiling solvents, for example from DMSO, DMF, etc.;

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

preferably, in the step, the reaction solvent may be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the protecting group removing condition may be selected from acidic conditions, for example, when the protecting group is selected from N-boc, palladium carbon, for example, when the protecting group is selected from N-cbz, piperidine, for example, when the protecting group is selected from N-fmoc;

(b3) carrying out condensation reaction on M-12 and 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 may be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature may be selected from 25 ℃ to 120 ℃, and the condensing agent may be selected from N, N ' -diisopropylcarbodiimide, benzotriazole-N, N ' -tetramethylurea hexafluorophosphate, 2- (7-azobenzotriazole) -N, N ' -tetramethylurea hexafluorophosphate, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate, and the like;

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

preferably, the conditions for removing the protecting group in said step may be selected from acidic conditions, for example, in the presence of palladium on carbon, when the protecting group is selected from N-boc, for example, in the presence of piperidine, when the protecting group is selected from N-cbz, for example, when the protecting group is selected from N-fmoc; the reaction solvent is selected from tetrahydrofuran, dichloromethane, methanol, toluene and the like;

(b5) carrying out condensation reaction on M-15 and 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 may be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature may be selected from 25 ℃ to 120 ℃, and the condensing agent may be selected from N, N ' -diisopropylcarbodiimide, benzotriazole-N, N ' -tetramethylurea hexafluorophosphate, 2- (7-azobenzotriazole) -N, N ' -tetramethylurea hexafluorophosphate, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate, and the like;

(b6) removing the protecting group from M-17 to obtain a target compound shown as a formula I;

preferably, the reaction solvent in the reaction is selected from tetrahydrofuran, dichloromethane, methanol, toluene and the like.

According to an embodiment of the present invention, in the second embodiment, the process for preparing formula I from M-9 comprises the steps of:

(c1) closing the ring of M-9 and N-1 under the condition of a catalyst to obtain N-2;

preferably, in said step, the catalyst may be selected from Oxone, sodium metabisulfite, etc., the reaction temperature may be selected from 25 ℃ to 140 ℃, and the reaction solvent may be selected from inert high boiling solvents, for example from DMSO, DMF, etc.;

(c2) selectively removing a protecting group from the N-2 to obtain N-3;

preferably, in the step, the reaction solvent may be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the protecting group removing condition may be selected from acidic conditions, for example, when the protecting group is selected from N-boc, palladium carbon, for example, when the protecting group is selected from N-cbz, piperidine, for example, when the protecting group is selected from N-fmoc;

(c3) carrying out condensation reaction on N-3 and 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 may be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature may be selected from 25 ℃ to 120 ℃, and the condensing agent may be selected from N, N ' -diisopropylcarbodiimide, benzotriazole-N, N ' -tetramethylurea hexafluorophosphate, 2- (7-azobenzotriazole) -N, N ' -tetramethylurea hexafluorophosphate, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate, and the like;

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

preferably, in the step, the reaction solvent may be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the protecting group removing condition may be selected from acidic conditions, for example, when the protecting group is selected from N-boc, palladium carbon, for example, when the protecting group is selected from N-cbz, piperidine, for example, when the protecting group is selected from N-fmoc;

(c5) carrying out condensation reaction on N-6 and 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 may be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature may be selected from 25 ℃ to 120 ℃, and the condensing agent may be selected from N, N ' -diisopropylcarbodiimide, benzotriazole-N, N ' -tetramethylurea hexafluorophosphate, 2- (7-azobenzotriazole) -N, N ' -tetramethylurea hexafluorophosphate, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate, and the like;

(c6) removing the protecting group from N-8 to obtain N-9;

preferably, in the step, the reaction solvent may be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the protecting group removing condition may be selected from acidic conditions, for example, when the protecting group is selected from N-boc, palladium carbon, for example, when the protecting group is selected from N-cbz, piperidine, for example, when the protecting group is selected from N-fmoc;

(c7) n-9 in inert organic solvent under the action of condensing agent and PG₂Carrying out condensation reaction on the protected chiral amino acid compound to obtain a compound N-10;

preferably, in the step, the inert organic solvent may be selected from acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, the temperature may be selected from 25 ℃ to 120 ℃, and the condensing agent may be selected from N, N ' -diisopropylcarbodiimide, benzotriazole-N, N ' -tetramethylurea hexafluorophosphate, 2- (7-azobenzotriazole) -N, N ' -tetramethylurea hexafluorophosphate, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate, and the like;

(c8) removing the protecting group from N-10 to obtain a target compound shown as a formula I;

preferably, in the step, the reaction solvent may be selected from tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the protecting group removing condition may be selected from acidic conditions, for example, when the protecting group is selected from N-boc, palladium carbon, for example, when the protecting group is selected from N-cbz, piperidine, for example, when the protecting group is selected from N-fmoc.

The present invention further provides a pharmaceutical composition comprising a compound of formula I as described herein and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof.

In some embodiments, the pharmaceutical compositions of the present invention further comprise a therapeutically effective amount of a compound of formula I of the present invention and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs thereof, or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.

The carrier in the pharmaceutical composition is "acceptable" in that it is compatible with (and preferably capable of stabilizing) the active ingredient of the composition and is not deleterious to the subject being treated. One or more solubilizing agents may be used as pharmaceutical excipients for the delivery of the active compound.

Accordingly, the present invention further provides the use of said compounds of formula I and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs thereof or pharmaceutically acceptable salts thereof in the preparation of an inhibitor of apoptosis proteins.

The invention further provides the use of a compound of formula I, and racemates, stereoisomers, tautomers, isotopic labels, nitric oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, in the manufacture of a medicament for the treatment of diseases or conditions due to IAP disorders.

The compounds of the invention are SMAC mimetics that are useful in the treatment of proliferative diseases, such as: various benign or malignant tumors (cancers), 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 (peripheral neuroma), bladder cancer, glioblastoma, melanoma, adrenocortical cancer, AIDS-related lymphoma, anal cancer, bladder cancer, meningioma, glioma, astrocytoma, breast cancer, cervical cancer, chronic myeloproliferative 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 bile duct cancer, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gestational trophoblastoma, hairy cell leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hypopharynx cancer, intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, laryngeal carcinoma, leukemia, neuroblastoma, melanoma, neuroblastoma, melanoma, bladder cancer, colon cancer, melanoma, Acute lymphoblastic leukemia, acute myeloid leukemia, lip cancer, oral cancer, liver cancer, male breast cancer, malignant mesothelioma, medulloblastoma, melanoma, Merkel (Merkel) cell carcinoma, metastatic squamous neck cancer, multiple myeloma and other plasma cell neoplasms, mycosis fungoides and Sezary syndrome (Sezary syndrome), myelodysplastic syndrome, nasopharyngeal cancer, neuroblastoma, non-small cell lung cancer, oropharyngeal cancer, bone cancer (including malignant fibrous histiocytoma of osteosarcoma and bone), epithelial ovarian cancer, germ cell tumor, low malignant potential tumor of ovary (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, neuroblastoma, melanoma, and other plasma cell neoplasms of the human body, and other tumors of the human body, Rhabdomyosarcoma, salivary gland carcinoma, skin cancer, small bowel cancer, soft tissue sarcoma, supratentorial primary neuroectodermal tumor, pinealoblastoma, testicular cancer, thymoma, thymus cancer, thyroid cancer, transitional cell carcinoma of renal pelvis and ureter, cancer of urethra, uterine cancer, vaginal cancer, vulval cancer, and Wilms's tumor and other childhood renal tumors.

Accordingly, the present invention further provides the use of a compound of formula I as described herein and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof or said pharmaceutical composition thereof in the manufacture of a medicament for the prevention and/or treatment of a tumor or tumor-related disorder as described above.

The methods of the invention may comprise administering a compound of the invention alone, as well as in combination with one or more other chemotherapeutic agents. Administration of multiple drugs can be simultaneous or sequential.

Interpretation of terms:

unless otherwise indicated, the definitions of groups and terms described in the specification and claims of the present application, including definitions thereof as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. The definitions of the groups and the structures of the compounds in such combinations and after the combination should fall within the scope of the present specification.

Where a range of numerical values is recited in the specification and claims herein, and where the range of numerical values is defined as an "integer," it is understood that the two endpoints of the range are recited and each integer within the range is recited. For example, "an integer of 0 to 6" should be understood to describe 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 may be described as "halogen" in the present specification.

The term "aliphatic hydrocarbon group" includes saturated or unsaturated, straight-chain or branched chain or cyclic hydrocarbon groups, the type of the aliphatic hydrocarbon group may be selected from alkyl, alkenyl, alkynyl and the like, the number of carbon atoms of the aliphatic hydrocarbon group is preferably 1 to 12, and may also be 1 to 10, and further preferably ranges from 1 to 6, and specifically may include but is not limited to the following groups: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, ethenyl, 1-propenyl, 2-propenyl, 1-methylethenyl, 1-butenyl, 1-ethylethenyl, 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 comprise one, two or more heteroatoms (or be construed as an optional insertion of heteroatoms into the aliphatic hydrocarbon group, optionally a C-C bond and a C-H bond). Suitable heteroatoms will be apparent to those skilled in the art and include, for example, sulfur, nitrogen, oxygen, phosphorus and silicon. The heteroatom containing aliphatic hydrocarbyl group may be selected from the following groups: (C)₁-C₆) Aliphatic hydrocarbyloxy, (C)₁-C₆) Aliphatic hydrocarbyl mercapto group, (C)₁-C₆) Aliphatic hydrocarbyloxy (C)₁-C₆) Aliphatic hydrocarbon group, (C)₁-C₆) Aliphatic hydrocarbyl mercapto group (C)₁-C₆) Aliphatic hydrocarbon group, N- (C)₁-C₃) Aliphatic hydrocarbyl amino group (C)₁-C₆) Aliphatic hydrocarbon group, N-di- (C)₁-C₃) Aliphatic hydrocarbyl amino group (C)₁-C₆) The aliphatic hydrocarbon group may be, for example, a methoxy group, an ethoxy group, a propoxy group, a butoxy group, a pentyloxy group, a methoxymethyl group, an ethoxymethyl group, a propoxymethyl group, a methoxyethyl group, a,Ethoxyethyl, propoxyethyl, methoxypropyl, ethoxypropyl, propoxypropyl, N-methylaminomethyl, N-methylaminoethyl, N-ethylaminoethyl, N-dimethylaminomethyl, N-dimethylaminoethyl, N-diethylaminoethyl; the "aliphatic hydrocarbon group" moiety contained in the other groups is as explained above.

The term "C_3-12Cycloalkyl "is understood to mean a saturated or unsaturated, monovalent monocyclic or bicyclic ring having 3 to 12 carbon atoms, preferably" C_3-10Cycloalkyl groups ". The term "C_3-10Cycloalkyl "is understood to mean a saturated or unsaturated monovalent monocyclic or bicyclic ring having 3,4, 5,6, 7, 8, 9 or 10 carbon atoms. Said C is_3-10Cycloalkyl groups may be monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or bicyclic, such as tetralin or decalin.

The term "3-12 membered heterocyclic group" means a saturated or unsaturated monovalent monocyclic or bicyclic ring containing 1 to 5 heteroatoms independently selected from N, O and S, the heteroatom-containing group having no aromaticity, said 3-12 membered heterocyclic group, preferably "3-10 membered heterocyclic group". The term "3-10 membered heterocyclyl" means a saturated monovalent monocyclic or bicyclic ring comprising 1-5, preferably 1-3 heteroatoms selected from N, O and S. The heterocyclic group may be attached to the rest of the molecule through any of the carbon atoms or nitrogen atom (if present). In particular, the heterocyclic group may include, but is not limited to: 4-membered rings such as azetidinyl, oxetanyl; 5-membered rings such as tetrahydrofuranyl, tetrahydrothienyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl; or a 7-membered ring such as diazepanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclyl group may be bicyclic, for example but not limited to a5, 5 membered ring, such as a hexahydrocyclopenta [ c ] pyrrol-2 (1H) -yl ring, or a5, 6 membered bicyclic ring, such as a hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl ring. The nitrogen atom containing ring may be partially unsaturated, i.e., it may contain one or more double bonds, such as, but not limited to, 2, 5-dihydro-1H-pyrrolyl, 4H- [1,3,4] thiadiazinyl, 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_6-20Aryl "is to be understood as preferably meaning a mono-, bi-or tricyclic hydrocarbon ring of monovalent or partially aromatic character having 6 to 20 carbon atoms, preferably" C_6-14Aryl ". The term "C_6-14Aryl "is to be understood as preferably meaning a mono-, bi-or tricyclic hydrocarbon ring having a monovalent or partially aromatic character with 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms (" C_6-14Aryl group "), in particular a ring having 6 carbon atoms (" C₆Aryl "), such as phenyl; or biphenyl, or is a ring having 9 carbon atoms ("C₉Aryl), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C₁₀Aryl radicals), such as tetralinyl, dihydronaphthyl or naphthyl, or rings having 13 carbon atoms ("C₁₃Aryl radicals), such as the fluorenyl radical, or a ring having 14 carbon atoms ("C)₁₄Aryl), such as anthracenyl.

The term "5-14 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: which has 5,6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and which contains 1 to 5, preferably 1 to 3 heteroatoms independently selected from N, O and S and, in addition, can be benzo-fused in each case. In particular, heteroaryl is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl and the like and their benzo derivatives, such as benzofuryl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl, isoquinolyl, and the like; or azocinyl, indolizinyl, purinyl and the like and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and the like.

Unless otherwise indicated, heterocyclyl or heteroaryl includes all possible isomeric forms thereof, e.g. positional isomers thereof. Thus, for some illustrative, non-limiting examples, pyridyl or pyridinylene includes pyridin-2-yl, pyridinylene-2-yl, pyridin-3-yl, pyridinylene-3-yl, pyridin-4-yl, and pyridinylene-4-yl; thienyl or thienylene includes thien-2-yl, thien-3-yl and thien-3-yl.

Depending on their molecular structure, the compounds of the invention may be chiral and may therefore exist in various enantiomeric forms. These compounds may thus be present in racemic or optically active form. The compounds of the invention or intermediates thereof may be separated into enantiomeric compounds by chemical or physical methods well known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from mixtures by reaction with optically active resolving agents. Examples of suitable resolving agents are optically active acids such as the R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g. N-benzoylproline or N-benzenesulfonylproline) or various optically active camphorsulphonic acids. The chromatographic enantiomeric resolution can also advantageously be carried out with the aid of optically active resolving agents, such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers, which are immobilized on silica gel. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example hexane/isopropanol/acetonitrile.

Those skilled in the art will appreciate that not all nitrogen-containing heterocycles can form N-oxides, as the nitrogen needs to have available lone pairs of electrons for oxidation to an oxynitride; those skilled in the art will recognize nitrogen-containing heterocycles that are capable of forming N-oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include the oxidation of heterocycles and tertiary amines with peroxy acids such as peracetic and m-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as t-butyl hydroperoxide, sodium perborate, and dioxiranes (dioxiranes) such as dimethyldioxirane. These methods for preparing N-oxides have been widely described and reviewed in the literature.

Pharmaceutically acceptable salts may be acid addition salts of the compounds of the invention having sufficient basicity, for example having a nitrogen atom in the chain or ring, for example with the following inorganic acids: for example hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid or nitric acid, or hydrogen sulfates, or acid addition salts with organic acids such as: such as 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, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic 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, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, Mandelic acid, ascorbic acid, glucoheptylic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid or thiocyanic acid.

In addition, another suitable pharmaceutically acceptable salt of a compound of the invention which is sufficiently acidic is an alkali metal salt (e.g., sodium or potassium salt), an alkaline earth metal salt (e.g., calcium or magnesium salt), an ammonium salt, or a salt with an organic base which affords a physiologically acceptable cation, such as a salt with: sodium ions, potassium ions, N-methylglucamine, dimethylglucamine, ethylglucamine, lysine, dicyclohexylamine, 1, 6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serinol, trihydroxymethylaminomethane, aminopropanediol, 1-amino-2, 3, 4-butanetriol. By way of example, the pharmaceutically acceptable salts include salts of the group-COOH with: sodium ion, potassium ion, calcium ion, magnesium ion, N-methylglucamine, dimethylglucamine, ethylglucamine, lysine, dicyclohexylamine, 1, 6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serinol, trihydroxymethylaminomethane, aminopropanediol, 1-amino-2, 3, 4-butanetriol.

In addition, the basic nitrogen-containing groups may be quaternized with the following agents: lower alkyl halides such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and diamyl sulfate; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl halides such as benzyl and phenethyl bromide, and the like. By way of example, pharmaceutically acceptable salts include hydrochloride, sulfate, nitrate, bisulfate, hydrobromide, acetate, oxalate, citrate, methanesulfonate, formate, or meglumine salts and the like.

Since the compound of the present invention may exist at a plurality of salt-forming sites, the "pharmaceutically acceptable salt" includes not only the salt formed at 1 salt-forming site among the compounds of the present invention but also the salt formed at 2,3 or all of the salt-forming sites among them. For this purpose, the molar ratio of the "pharmaceutically acceptable salt" of the compound of formula (I) to the cation of the acid (anion) or base required for salt formation may vary within wide limits, and may be, for example, 4:1 to 1:4, such as 3:1, 2:1, 1:2, 1:3, etc.

According to the present invention, the pharmaceutically acceptable anion includes anions selected from the group consisting of those generated by ionization of inorganic 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, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic 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, maleic, Fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptonic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid or thiocyanic acid.

Depending on the position and nature of the various substituents, the compounds of the present invention may also contain one or more asymmetric centers. Asymmetric carbon atoms may exist in either the (R) or (S) configuration, with only one asymmetric center yielding a racemic mixture and multiple asymmetric centers yielding a diastereomeric mixture. In some cases, asymmetry may also exist due to hindered rotation about a particular bond, for example, the central bond connects two substituted aromatic rings of a particular compound. Also, the substituents may exist in cis or trans isomeric forms.

The compounds of the invention also include all possible stereoisomers of each, either as a single stereoisomer or as any mixture of said stereoisomers (e.g. the R-or S-isomers, or the E-or Z-isomers) in any proportion. Separation of individual stereoisomers (e.g. individual enantiomers or individual diastereomers) of the compounds of the invention may be achieved by any suitable prior art method (e.g. chromatography, particularly, for example, chiral chromatography).

The term "tautomer" refers to an isomer of a functional group resulting from the rapid movement of an atom in two positions in a molecule. The compounds of the invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer often result in a mixture whose physicochemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; whereas in phenol the enol type predominates. The present invention encompasses all tautomeric forms of the compounds.

In the present invention, reference to compounds also includes isotopically-labeled compounds, which are identical to those shown in formula I, but wherein one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of H, C, N, O, S, F and Cl, such as²H、³H、¹³C、¹¹C、¹⁴C、¹⁵N、¹⁸O、¹⁷O、³²P、³⁵S、¹⁸F and³⁶and (4) Cl. Compounds of the present invention, prodrugs thereof, or pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labeled compounds of the present invention, for example, by incorporation of a radioactive isotope (such as³H and¹⁴C) the compounds of (a) are useful in drug and/or substrate tissue distribution assays. Tritium (i.e. tritium³H) And carbon 14 (i.e.¹⁴C) Isotopes are particularly preferred for their ease of preparation and detectability. Again, with heavier isotopes such as deuterium (i.e. deuterium)²H) Substitutions may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life)Phase or reduced dosage requirements) and may therefore be preferred in certain circumstances. The compounds of the invention as claimed may be particularly limited to replacement with deuterium or tritium. Furthermore, the absence of hydrogen in the substituents indicating the term deuterium or tritium alone is not meant to exclude deuterium or tritium, but may equally well comprise deuterium or tritium.

The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound of the present invention sufficient to effect the intended use, including but not limited to the treatment of a disease 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 and disease condition being treated, such as the weight and age of the subject, the severity of the disease condition and the mode of administration, etc., can be readily determined by one of ordinary skill in the art. The specific dosage will vary depending on the following factors: the particular compound selected, the dosage regimen to be followed, whether to administer it in combination with other compounds, the timing of administration, the tissue to be administered and the physical delivery system carried.

The term "solvate" is those forms of the compounds of the present invention which form complexes in the solid or liquid state by coordination with solvent molecules. Hydrates are a particular form of solvates in which the coordination is with water. In the present invention, the preferred solvate is a hydrate. Further, pharmaceutically acceptable solvates (hydrates) of the compounds of general formula I according to the invention refer to co-crystals and clathrates of compound I with one or more molecules of water or other solvents in stoichiometric amounts. Solvents that may be used for the solvate include, but are not limited to: water, methanol, ethanol, ethylene glycol and acetic acid.

The term "prodrug", or "prodrug" refers to a compound that is converted in vivo to a compound of the general formula or a particular compound. Such conversion is effected by hydrolysis of the prodrug in the blood or by enzymatic conversion to the parent structure in the blood or tissue. The prodrugs of the present invention may be esters, and esters useful as prodrugs in the present invention are phenyl esters, aliphatic (C1-24) esters, acyloxymethyl esters, carbonates, carbamates and amino acid esters. For example, a compound of the present invention comprises a hydroxy/carboxy group, i.e., it may be acylated to provide the compound in prodrug form. Other prodrug forms include phosphate esters, such as those obtained by phosphorylation of a hydroxyl group on the parent.

Advantageous effects

1) The compound adopts a novel general formula I structure, has better binding affinity for XIAP, cIAP1 and cIAP2 proteins, has better IAP inhibitory activity, and has the inhibitory effect of some compounds which is obviously better than that of positive control drugs;

2) the compound has better inhibition effect on cell growth in MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines, and the inhibition effect of some compounds is obviously better than that of positive control drugs.

Detailed Description

The technical solution of the present invention will be further described in detail with reference to specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.

Unless otherwise indicated, 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 embodiments, the processes for obtaining chiral compounds include the step of preparative separation and purification using a chiral column.

Example 1: preparation of benzyloxycarbonyl (2S) -2- [ (1, 3-dioxoisoindol-2-yl) methyl ] pyrrolidine-1-carboxylic acid ester

93.5g (0.398mol) of N-Cbz-L-prolinol, 70.2g (0.478mol) of phthalimide and 104.3g (0.398mol) of triphenylphosphine are put into a 2L three-port reaction bottle, tetrahydrofuran (1.3L) is added, the system is cooled to 0 ℃ under the protection of nitrogen, and DIAD80.4g (0.398mol) is dropwise added at 0 ℃. The system was then warmed to room temperature and stirred for 16 h, TLC checked for reaction completion, quenched with 400mL water, stirred for an additional 30 min and extracted with ethyl acetate (300 mL. times.3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:40) to give benzyloxycarbonyl (2S) -2- [ (1, 3-dioxoisoindol-2-yl) methyl ] pyrrolidine-1-carboxylic acid ester (98.5g, 68%).

Example 2: preparation of benzyloxycarbonyl (2S) -2- (aminomethyl) pyrrolidine-1-carboxylic acid ester

Carbobenzoxy (2S) -2- [ (1, 3-dioxo-isoindol-2-yl) methyl ] pyrrolidine-1-carboxylic acid ester 65.5g (0.18mol) was put in a 1L reaction flask, 450mL of ethanol was added, the mixture was heated to 80 ℃ and 17g (0.45mol, 85%) of hydrazine hydrate was added dropwise thereto. Then stirred at 80 ℃ for 12 hours and the reaction was complete by TLC. Cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give benzyloxycarbonyl (2S) -2- (aminomethyl) pyrrolidine-1-carboxylic acid ester (38.7g, 92%) which was used directly in the next reaction.

Example 3: preparation of carbobenzoxy (2S) -2- [ (4-fluoro-2-nitro-anilino) methyl ] pyrrolidine-1-carboxylic acid ester

35g (0.15mol) of benzyloxycarbonyl (2S) -2- (aminomethyl) pyrrolidine-1-carboxylic acid ester was put in a 500mL reaction flask, 350mL of acetonitrile was added thereto, 41.4g (0.3mol) of potassium carbonate was added thereto, and 26.2g (0.17mol) of 1, 4-difluoro-2-nitrobenzene was added thereto. Under the protection of nitrogen, the temperature is raised to 80 ℃, the mixture is stirred for 1 hour, the TLC detects that the reaction is complete, water is added for 200mL for quenching, and ethyl acetate (200mL multiplied by 3) is extracted. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether: 1:40) to give benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-nitro-anilino) methyl ] pyrrolidine-1-carboxylic acid ester (49.2g, 88%).

Example 4: preparation of carbobenzoxy (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] pyrrolidine-1-carboxylic acid ester

Carbobenzoxy (2S) -2- [ (4-fluoro-2-nitro-anilino) methyl ] pyrrolidine-1-carboxylate 49.2g (0.13mol) was taken in a 1L reaction flask, 400mL of acetic acid was added, 135g of iron powder was added thereto, stirring was performed at room temperature for 16 hours, TLC detected reaction was complete, filtration was performed, the filter cake was rinsed with ethyl acetate, the filtrate was concentrated under reduced pressure, 300mL of water was added, ethyl acetate (200 mL. times.3) was extracted, the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. This gave benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] pyrrolidine-1-carboxylic acid ester (43.5g, 96%) which was used directly in the next reaction.

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

30g (87.4mmol) of carbobenzoxy (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] pyrrolidine-1-carboxylate is put into a 500mL reaction bottle, 300mL of 4N hydrochloric acid is added, 20g (0.262mol) of acetic alcohol is added, the mixture is heated to 100 ℃ and stirred for 6 hours, TLC detects the reaction is completed, the temperature is reduced to 0 ℃, saturated sodium bicarbonate solution is added to adjust the pH to be 8-9, the mixture is filtered, a filter cake is washed by methyl tert-butyl ether and dried, and (S) -benzyl 2- ((5-fluoro-2- (hydroxymethyl) -1H-benzo [ D ] imidazol-1-yl) methyl) pyrrolidine-1-carboxylate (29g, 87%) is obtained and is directly used for the next reaction.

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

Taking (S) -benzyl 2- ((5-fluoro-2- (hydroxymethyl) -1H-benzo [ D)]Imidazole-1-yl) methyl) pyrrolidine-1-carboxylic acid ester 20g (52.2mmol) is put into a 500mL reaction bottle, 300mL dichloromethane is added, the temperature is reduced to 0 ℃ under the protection of nitrogen, another 26.6g (62.6mmol) of dessimidine reagent is added, the temperature is restored to room temperature, the mixture is stirred for 2 hours, the TLC detection reaction is completed, the temperature is reduced to 0 ℃, NaHCO is used for detecting the reaction completion₃/Na₂S₂O₃(1:1) the saturated solution (200mL x 2) was washed, the aqueous phase was extracted with dichloromethane (200mL x 2), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:5) to give (S) -benzyl 2- ((5-fluoro-2-formyl-1H-benzo [ D)]Imidazol-1-yl) methyl) pyrrolidine-1-carboxylic acid ester (18.3g, 92%).

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

15g (39.4mmol) of (S) -benzyl 2- ((5-fluoro-2-formyl-1H-benzo [ D ] imidazol-1-yl) methyl) pyrrolidine-1-carboxylate, 16.2g (47.2mmol) of benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] pyrrolidine-1-carboxylate were taken in a 250mL reaction flask, 150mL of N, N-dimethylformamide was added, 28.5g (0.15mol) of sodium metabisulfite was added, nitrogen was used for protection, the mixture was stirred under microwave conditions at reflux for 1 hour, the reaction was checked by TLC to completion, the flask was returned to room temperature, 500mL of water was added, ethyl acetate (200 mL. times.3) was extracted, the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:5) to give (2S,2'S) -dibenzyl 2,2' - ((5,5 '-difluoro-1H, 1' H- [2,2 '-dibenzylo [ d ] imidazole ] -1,1' -diyl) bis (methylene)) bis (pyrrolidine-1-carboxylate) (22.5g, 81%).

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

Taking 10g (14.2mmol) of (2S,2'S) -dibenzyl 2,2' - ((5,5 '-difluoro-1H, 1' H- [2,2 '-dibenzyl [ D ] imidazole ] -1,1' -diyl) bis (methylene)) bis (pyrrolidine-1-carboxylate) in a 250mL reaction bottle, adding 100mL of methanol, adding 1g of palladium carbon into the reaction bottle, stirring at room temperature for 12 hours under a hydrogen atmosphere, detecting the completion of the reaction by TLC, filtering, and concentrating the filtrate to obtain 5,5 '-difluoro-1, 1' -bis ((S) -pyrrolidine-2-ylmethyl) -1H,1 'H-2, 2' -dibenzo [ D ] imidazole (5.7g, 93% crude product) which is directly used for the next reaction.

Example 9: preparation of 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

2.2g (10mmol) of N-Boc-L valine was taken in a 100mL reaction flask, 20mL of N-dimethylformamide was added thereto, 2g (20mmol) of N-methylmorpholine and 2g (15mmol) of HATU5.7g were added thereto, and stirred at room temperature for 30 minutes, then 2g (4.6mmol) of 5,5 '-difluoro-1, 1' -bis ((S) -pyrrolidin-2-ylmethyl) -1H,1 'H-2, 2' -dibenzo [ D ] imidazole was dissolved in 20mL of N, N-dimethylformamide and added dropwise to the above system, stirred at room temperature for 3 hours, TLC checked for completion of the reaction, 200mL of water was added, ethyl acetate (200 mL. times.3) was extracted, the combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:3) to give 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 (2.5g, 65%).

Example 10: preparation of (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 (pyrrolidin-2, 1-diyl)) bis (2-amino-3-methylbutan-1-one)

1.0g (1.2mmol) of 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 was taken in a 100mL reaction flask, 20mL of an ethanol hydrochloride solution was added thereto, the mixture was stirred at room temperature for 1 hour, the reaction was completed by TLC, the solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 0.8g of a colorless oily liquid, which was used directly in the next reaction.

Example 11: preparation of 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 (azepindiyl)) bis (1-oxopropane-2, 1-diyl)) bis (methylcarbamate)

0.58g (2.88mmol) of N-Boc-N-methyl-L-alanine was taken in a 100mL reaction flask, 20mL of N-dimethylformamide was added thereto, 0.61g (6.0mmol) of N-methylmorpholine and 0.37g (3.6mmol) of HATU1 were added thereto, and the mixture was stirred at room temperature for 30 minutes, and then 0.8g (crude, 1.2mmol) of (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 (pyrrolidin-2, 1-diyl) bis (2-amino-3-methylbutan-1-one) was taken and dissolved in 20mL of N-dimethylformamide, the mixture was added dropwise to the above system, stirred at room temperature for 3 hours, TLC checked for completion of the reaction, 200mL of water was added, ethyl acetate (200 mL. times.3) was extracted, and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by a silica gel column (eluent was ethyl acetate: petroleum ether ═ 1:3) to give 0.68g (53.5%) of 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)) bis (azepinyl)) bis (1-oxopropane-2, 1-diyl)) bis (methylcarbamate).

Example 12: preparation of (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)

Taking 0.68g (0.68mmol) of 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 (azepinyl)) bis (1-oxopropane-2, 1-diyl)) bis (carbamate) in a 100mL reaction flask, adding 20mL of an ethanol hydrochloride solution, stirring at room temperature for 1 hour, detecting by TLC that the reaction is complete, evaporating the solvent and excess hydrogen chloride under reduced pressure, adding 100mL of water, the pH was adjusted to 8 with saturated sodium bicarbonate, extracted with ethyl acetate (100mL × 2), and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by a silica gel column (eluent was ethyl acetate: petroleum ether ═ 1:2) to give 312mg (57%) of (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).

¹H-NMR(400MHz,d₆-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

The following example compounds were synthesized according to the synthetic methods of examples 1-12 and the first synthetic scheme:

example 13:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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),2.24-2.02(m,10H),1.38-1.26(m,6H),0.94-0.87(d,6H),[M+H]⁺:m/z＝777.8

example 18:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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: preparation of (S) -benzyl 2- ((1'- ((S) -1- (tert-butoxycarbonyl) pyrrolidin-2-yl) methyl) -5,5' -difluoro-1H, 1'H- [2,2' -bibenzo [ d ] imidazol ] -1-yl) methyl) pyrrolidine-1-carboxylic acid ester

15g (39.4mmol) of (S) -benzyl 2- ((5-fluoro-2-formyl-1H-benzo [ D ] imidazol-1-yl) methyl) pyrrolidine-1-carboxylate, 14.5g (47.2mmol) of tert-butoxycarbonyl (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] pyrrolidine-1-carboxylate, were taken in a 250mL reaction flask, 150mL of N, N-dimethylformamide was added, 28.5g (0.15mol) of sodium metabisulfite was added, nitrogen was protected, the mixture was stirred under microwave conditions at reflux for 1 hour, the reaction was checked by TLC to completion, the flask was returned to room temperature, 500mL of water was added, ethyl acetate (200 mL. times.3) was extracted, the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:5) to give (S) -benzyl 2- ((1'- ((S) -1- (tert-butoxycarbonyl) pyrrolidin-2-yl) methyl) -5,5' -difluoro-1H, 1'H- [2,2' -biphenylo [ d ] imidazol ] -1-yl) methyl) pyrrolidine-1-carboxylic acid ester (20.6g, 78%).

Example 27: preparation of (S) -benzyl 2- ((5,5 '-difluoro-1' - ((S) -pyrrolidin-2-ylmethyl) -1H,1'H- [2,2' -biizoyl [ d ] imidazol ] -1-yl) methyl) pyrrolidine-1-carboxylate

Taking 10g (14.9mmol) of (S) -benzyl 2- ((1'- ((S) -1- (tert-butoxycarbonyl) pyrrolidin-2-yl) methyl) -5,5' -difluoro-1H, 1'H- [2,2' -biphenyl [ d ] imidazole ] -1-yl) methyl) pyrrolidine-1-carboxylic acid ester into a 250mL reaction bottle, adding 120mL of hydrochloric acid ethanol solution, stirring for 1 hour at room temperature, detecting the reaction by TLC, evaporating the solvent and excessive hydrogen chloride under reduced pressure to obtain 8.6g of colorless oily liquid, and directly using the colorless oily liquid in the next reaction. And directly used for the next reaction.

Example 28: preparation of (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' -dibenzo [ D ] imidazole ] -1-yl) methyl) pyrrolidine-1-carboxylate

To a 250mL reaction flask was added 3.9g (17.9mmol) of N-Boc-L valine, 80mL of N-dimethylformamide, and to this was added 4.5g (44.7mmol) of N-methylmorpholine and 4.5g (22.4mmol) of HATU8.5g (22.4mmol), and the mixture was stirred at room temperature for 30 minutes, then 8.6g (crude, 14.9mmol) of (S) -benzyl 2- ((5,5 '-difluoro-1' - ((S) -pyrrolidin-2-ylmethyl) -1H,1'H- [2,2' -biimidazole ] -1-yl) methyl) pyrrolidine-1-carboxylate was dissolved in 40mL of N-dimethylformamide, and the mixture was added dropwise to the above system, stirred at room temperature for 3 hours, TLC was checked for completion of the reaction, 600mL of water was added, ethyl acetate (200 mL. times.3) was extracted, and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:3) to give (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' -dibenzo [ D ] imidazol ] -1-yl) methyl) pyrrolidine-1-carboxylate 8.1g (70%).

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

Taking 8.1g (10.5mmol) of (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' -dibenzo [ D ] imidazole ] -1-yl) methyl) pyrrolidine-1-carboxylate into a 250mL reaction bottle, adding 100mL of methanol, further taking 0.8g of palladium carbon, adding the palladium carbon into the reaction bottle, stirring the mixture at room temperature for 12 hours under a hydrogen atmosphere, detecting complete reaction by TLC, filtering, concentrating the filtrate to obtain tert-butyl ((R) -1- ((S) -2- ((5,5 '-difluoro-1' - ((S) -pyrrolidin-2-ylmethyl) -1H, 6.7g of crude 1'H- [2,2' -dibenzo [ d ] imidazol ] -1-yl) methyl) pyrrolidin-1-yl) -3-methyl-1-oxobutan-2-yl) carbamate was used directly in the next reaction.

Example 30: preparation of di-tert-butyl ((2R,2' R) -2-amino-1- ((S) -2- ((1' - ((S) -1- ((R) -2-aminopropanoyl) 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)

0.35g (1.89mmol) of N-Boc-D-alanine was taken in a 100mL reaction flask, 20mL of N, N-dimethylformamide was added thereto, 0.32g (3.14mmol) of N-methylmorpholine and 0.90g (2.36mmol) of HATU were added thereto, and stirred at room temperature for 30 minutes, then 1.0g (crude, 1.57mmol) of 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 was taken and dissolved in 20mL of N, N-dimethylformamide, the mixture was added dropwise to the above system, stirred at room temperature for 3 hours, TLC checked for completion of the reaction, 600mL of water was added, extracted with ethyl acetate (100 mL. times.3), and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:5) to give 0.91g (72%) of 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).

Example 31: preparation of (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

0.91g (1.1mmol) of di-tert-butyl ((2R,2' R) -2-amino-1- ((S) -2- ((1' - ((S) -1- ((R) -2-aminopropanoyl) 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) was taken in a 100mL reaction flask, 20mL of an ethanol hydrochloride solution was added, and the mixture was stirred at room temperature for 1 hour, the TLC check was done and the solvent and excess hydrogen chloride were distilled off under reduced pressure to give 0.7g of a colorless oily liquid which was used directly in the next reaction.

Example 32: preparation of 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' -biizo[ D ] imidazol ] -1-yl) methyl) pyrrolidin-1-yl) -3-methyl-1-oxobutan-2-yl) -2- (methylamino) propionamide-bis (methylcarbamate)

0.54g (2.64mmol) of N-Boc-N-methyl-L-alanine was placed in a 100mL reaction flask, 20mL of N, N-dimethylformamide was added thereto, 0.56g (5.5mmol) of N-methylmorpholine and 0.3 g (3.3mmol) of HATU1.3g (3.3mmol) were added thereto, and the mixture was stirred at room temperature for 30 minutes, followed by dissolving 0.7g (1.1mmol) of (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 in 20mL of N, n-dimethylformamide was added dropwise to the above system, stirred at room temperature for 3 hours, the reaction was complete by TLC, 200mL of water was added, extracted with ethyl acetate (200mL × 3), and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified on a silica gel column (eluent ethyl acetate: petroleum ether ═ 1:3) to give di-tert-butyl ((2R,2' R) - (R) -N- ((R) -1- ((S) -2- ((5,5' -difluoro-1 ' - ((S) -1- ((R) -2- (methylamino) alanyl) pyrrolidin-2-yl) methyl) -1H,1' H- [2,2' -biszo[ D ] imidazol ] -1-yl) methyl) pyrrolidin-1-yl) -3-methyl-1-oxobutan-2-yl) -2- (methylamino) propionamide-bis (methylcarbamate) 0.66g (61%).

Example 33: preparation of (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' -biizo[ D ] imidazol ] -1-yl) methyl) pyrrolidin-1-yl) -3-methyl-1-oxobutan-2-yl) -2- (methylamino) propionamide

Taking 0.66g (0.68mmol) of 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' -biizo[ D ] imidazol ] -1-yl) methyl) pyrrolidin-1-yl) -3-methyl-1-oxobutan-2-yl) -2- (methylamino) propionamide-bis (carbamate) into a 100mL reaction flask, adding 20mL of an ethanol hydrochloride solution, after stirring at room temperature for 1 hour, the reaction was complete by TLC, the solvent and excess hydrogen chloride were evaporated under reduced pressure, 100mL of water was added, the pH was adjusted to 8 with saturated sodium bicarbonate, ethyl acetate was extracted (100mL × 2), and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:2) to give 513mg (62%) of (R) -N- ((R) -1- ((S) -2- ((5,5 '-difluoro-1' - ((S) -1- ((R) -2- (methylamino) alanyl) pyrrolidin-2-yl) methyl) -1H,1'H- [2,2' -bizoio [ D ] imidazol ] -1-yl) methyl) pyrrolidin-1-yl) -3-methyl-1-oxobutan-2-yl) -2- (methylamino) propionamide.

¹H-NMR(400MHz,d₆-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

The following compounds of examples were synthesized according to the synthesis methods of examples 26 to 33 and the second synthesis scheme:

example 34:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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-carboxylic acid ester

80g (0.319mol) of N-Cbz-4-hydroxy-L-prolinol, 56.2g (0.382mol) of phthalimide and 83.6g (0.319mol) of triphenylphosphine are put into a 2L three-mouth reaction bottle, tetrahydrofuran (1.3L) is added, the system is cooled to 0 ℃ under the protection of nitrogen, and DIAD64.4g (0.319mol) is dropwise added at 0 ℃. The system was then warmed to room temperature and stirred for 16 h, TLC checked for reaction completion, quenched with 400mL water, stirred for an additional 30 min and extracted with ethyl acetate (300 mL. times.3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:20) to give benzyloxycarbonyl (2S) -2- [ (1, 3-dioxoisoindol-2-yl) methyl ] -4-hydroxypyrrolidine-1-carboxylic acid ester (64.2g, 53%).

Example 43: preparation of benzyloxycarbonyl (2S) -2- (aminomethyl) -4-hydroxypyrrolidine-1-carboxylic acid ester

64.2g (0.17mol) of benzyloxycarbonyl (2S) -2- [ (1, 3-dioxoisoindol-2-yl) methyl ] -4-hydroxypyrrolidine-1-carboxylate was put into a 1L reaction flask, 450mL of ethanol was added thereto, the mixture was heated to 80 ℃ and 17g (0.45mol, 85%) of hydrazine hydrate was added dropwise thereto. Then stirred at 80 ℃ for 12 hours and the reaction was complete by TLC. Cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give benzyloxycarbonyl (2S) -2- (aminomethyl) -4-hydroxypyrrolidine-1-carboxylic acid ester (38.3g, 90%) which was used directly in the next reaction.

Example 44: preparation of benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-nitro-anilino) methyl ] -4-hydroxypyrrolidine-1-carboxylic acid ester

37.5g (0.15mol) of benzyloxycarbonyl (2S) -2- (aminomethyl) -4-hydroxypyrrolidine-1-carboxylate was put into a 500mL reaction flask, 350mL of acetonitrile was added thereto, 41.4g (0.3mol) of potassium carbonate was added thereto, and 26.2g (0.17mol) of 1, 4-difluoro-2-nitrobenzene was added thereto. Under the protection of nitrogen, the temperature is raised to 80 ℃, the mixture is stirred for 1 hour, the TLC detects that the reaction is complete, water is added for 200mL for quenching, and ethyl acetate (200mL multiplied by 3) is extracted. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:20) to give benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-nitro-anilino) methyl ] -4-hydroxypyrrolidine-1-carboxylic acid ester (46.7g, 80%).

Example 45: preparation of benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] -4-hydroxypyrrolidine-1-carboxylic acid ester

The carbobenzoxy (2S) -2- [ (4-fluoro-2-nitro-anilino) methyl ] -4-hydroxypyrrolidine-1-carboxylate 45g (0.12mol) was placed in a 1L reaction flask, 400mL of acetic acid was added, 135g of iron powder was added thereto, the mixture was stirred at room temperature for 16 hours, TLC detected for completion of the reaction, filtered, the filter cake was rinsed with ethyl acetate, the filtrate was concentrated under reduced pressure, 300mL of water was added, ethyl acetate (200 mL. times.3) was extracted, the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. This gave benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] -4-hydroxypyrrolidine-1-carboxylic acid ester (38.6g, 93%) which was used directly in the next reaction.

Example 46: preparation of (2S) -benzyl 2- ((5-fluoro-2- (hydroxymethyl) -1H-benzo [ d ] imidazol-1-yl) methyl) -4-hydroxypyrrolidine-1-carboxylate

31.3g (87.4mmol) of benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] -4-hydroxypyrrolidine-1-carboxylic acid ester was taken in a 500mL reaction flask, adding 300mL of 4N hydrochloric acid, adding 20g (0.262mol) of acetic alcohol, heating to 100 ℃, stirring for 6 hours, the reaction was checked by TLC to completion, cooled to 0 ℃, saturated sodium bicarbonate solution was added dropwise to adjust pH 8-9, filtered, the filter cake was washed with methyl tert-butyl ether and dried to give (2S) -benzyl 2- ((5-fluoro-2- (hydroxymethyl) -1H-benzo [ d ] imidazol-1-yl) methyl) -4-hydroxypyrrolidine-1-carboxylate (28.9g, 83%) which was used directly in the next reaction.

Example 47: preparation of (2S) -benzyl 2- ((5-fluoro-2-formyl-1H-benzo [ d ] imidazol-1-yl) methyl) -4-hydroxypyrrolidine-1-carboxylate ester

Taking (2S) -benzyl 2- ((5-fluoro-2- (hydroxymethyl) -1H-benzo [ d)]Imidazole-1-yl) methyl) -4-hydroxypyrrolidine-1-carboxylic acid ester 20.8g (52.2mmol) is put into a 500mL reaction bottle, 300mL dichloromethane is added, the temperature is reduced to 0 ℃ under the protection of nitrogen, and then 26.6g (62.6mmol) of dessimidine reagent is added, the temperature is restored to room temperature, the stirring is carried out for 2 hours, the TLC detection reaction is completed, the temperature is reduced to 0 ℃, NaHCO is used for detecting the reaction₃/Na₂S₂O₃(1:1) saturated solution(200mL x 2) washing, aqueous phase was extracted with dichloromethane (200mL x 2), combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:5) to give (2S) -benzyl 2- ((5-fluoro-2-formyl-1H-benzo [ d)]Imidazol-1-yl) methyl) -4-hydroxypyrrolidine-1-carboxylate ester (18.7g, 90%).

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

Taking 15g (37.8mmol) of (2S) -benzyl 2- ((5-fluoro-2-formyl-1H-benzo [ d ] imidazol-1-yl) methyl) -4-hydroxypyrrolidine-1-carboxylate ester, 16.3g (45.3mmol) of carbobenzoxy (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] -4-hydroxypyrrolidine-1-carboxylate in a 250mL reaction flask, adding 150mLN, N-dimethylformamide, adding 28.5g (0.15mol) of sodium metabisulfite, stirring under nitrogen and microwave conditions for 1 hour at reflux, detecting by TLC that the reaction is complete, returning to room temperature, adding 500mL of water, extracting with ethyl acetate (200 mL. times.3), washing the combined organic layers with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:5) to give (5S,5'S) -dibenzyl 5,5' - ((5,5 '-difluoro-1H, 1' H- [2,2 '-dibenzylo [ d ] imidazole ] -1,1' -diyl) bis (methylene)) bis (3-hydroxypyrrolidine-1-carboxylate) (21.7g, 78%).

Example 49: preparation of (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)

20g (27.2mmol) of (5S,5'S) -dibenzyl 5,5' - ((5,5 '-difluoro-1H, 1' H- [2,2 '-dibenzylo [ d ] imidazole ] -1,1' -diyl) bis (methylene)) bis (3-hydroxypyrrolidine-1-carboxylate) was put in a 250mL reaction flask, 100mL of methanol was added thereto, 1g of palladium on charcoal was further added thereto under a hydrogen atmosphere, and the mixture was stirred at room temperature for 12 hours, and the reaction was detected by TLC to be complete, followed by filtration and concentration of the filtrate 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) was used directly in the next reaction.

Example 50: preparation of 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-oxobutane-2, 1-diyl)) dicarbamate

2.2g (10.3mmol) of N-Boc-L valine was taken in a 100mL reaction flask, 20mL of N-dimethylformamide was added thereto, 2.2g (21.5mmol) of N-methylmorpholine and 2.9 g (12.9mmol) of HATU4 were added thereto, and stirred at room temperature for 30 minutes, then 2g (4.3mmol) of (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) was taken and dissolved in 20mL of N-dimethylformamide, and added dropwise to the above system, stirred at room temperature for 3 hours, TLC detection reaction was complete, 200mL of water was added, ethyl acetate (200 mL. times.3) was extracted, the combined organic layers were washed with brine and dried without water, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:3) to give 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-oxobutane-2, 1-diyl)) dicarbamate (2.2g, 58%).

Example 51: preparation of (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-methylbutan-1-one)

1.0g (1.2mmol) of 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-oxobutane-2, 1-diyl)) dicarbamate was taken in a 100mL reaction flask, 20mL of an ethanol solution of hydrochloric acid was added, stirring was carried out at room temperature for 1 hour, TLC detection was carried out for completion of the reaction, and the solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 0.8g of a colorless oily liquid, which was used directly in the next reaction.

Example 52: preparation of 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 (azepindiyl)) bis (1-oxopropane-2, 1-diyl)) bis (methyl carbamate)

0.58g (2.88mmol) of N-Boc-N-methyl-L-alanine was taken in a 100mL reaction flask, 20mL of N-dimethylformamide was added thereto, 0.61g (6.0mmol) of N-methylmorpholine and 0.37g (3.6mmol) of HATU1 were added thereto, and the mixture was stirred at room temperature for 30 minutes, and then 0.8g (crude, 1.2mmol) of (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) was taken and dissolved in 20mL of N-dimethylformamide, the mixture was added dropwise to the above system, stirred at room temperature for 3 hours, TLC checked for completion of the reaction, 200mL of water was added, ethyl acetate (200 mL. times.3) was extracted, and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:3) to give 0.57g (46%) of 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-oxobutane-2, 1-diyl)) bis (azepindiyl)) bis (1-oxopropane-2, 1-diyl)) bis (methyl carbamate).

Example 53: preparation of (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)) bis (3-methyl-1-oxobutane-2, 1-diyl)) bis (2- (methylamino) propionamide)

Taking 0.57g (0.55mmol) of 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 (azepinyl)) bis (1-oxopropane-2, 1-diyl)) bis (methyl carbamate) in a 100mL reaction flask, adding 20mL of hydrochloric acid ethanol solution, stirring at room temperature for 1 hour, detecting the reaction completion by TLC, reducing the pressure of the solvent and the excess hydrogen chloride, 100mL of water was added, pH was adjusted to 8 with saturated sodium bicarbonate, extracted with ethyl acetate (100mL × 2), and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by a silica gel column (eluent was ethyl acetate: petroleum ether ═ 1:2) to give 300mg (65%) of (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)) bis (3-methyl-1-oxobutane-2, 1-diyl)) bis (2- (methylamino) propionamide).

¹H-NMR(400MHz,d₆-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

The following compounds of examples were synthesized according to the synthesis methods of examples 42 to 53 and the first synthesis scheme:

example 54:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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-carboxylic acid ester

100g (0.26mol) of carbobenzoxy (2S) -2- [ (1, 3-dioxoisoindol-2-yl) methyl ] -4-hydroxypyrrolidine-1-carboxylate is taken to be put into a 1L three-mouth reaction bottle, dichloromethane (600mL) is added, the system is cooled to-78 ℃ under the protection of nitrogen, DAST63.6g (0.41mol) is dropwise added at-78 ℃, after dropwise addition is finished, the system is stirred for 3 hours at-78 ℃, then the system is heated to room temperature and stirred for 16 hours, TLC detection reaction is complete, the reaction liquid is poured into 800mL of saturated sodium bicarbonate solution at 0 ℃ to be quenched, and dichloromethane (300mL multiplied by 3) is extracted. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:40) to give benzyloxycarbonyl (2S) -2- [ (1, 3-dioxoisoindol-2-yl) methyl ] -4-fluoropyrrolidine-1-carboxylic acid ester (22g, 22%).

Example 70: preparation of benzyloxycarbonyl (2S) -2- (aminomethyl) -4-fluoropyrrolidine-1-carboxylic acid ester

22g (57.6mmol) of benzyloxycarbonyl (2S) -2- [ (1, 3-dioxoisoindol-2-yl) methyl ] -4-fluoropyrrolidine-1-carboxylic acid ester was put in a 1L reaction flask, 450mL of ethanol was added thereto, the mixture was heated to 80 ℃ and 5.7g (0.14mol, 85%) of hydrazine hydrate was added dropwise thereto. Then stirred at 80 ℃ for 12 hours and the reaction was complete by TLC. Cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give benzyloxycarbonyl (2S) -2- (aminomethyl) -4-hydroxypyrrolidine-1-carboxylic acid ester (13.5g, 93%) which was used directly in the next reaction.

Example 71: preparation of benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-nitro-anilino) methyl ] -4-fluoropyrrolidine-1-carboxylic acid ester

Into a 500mL reaction flask was taken 13.5g (53.6mmol) of benzyloxycarbonyl (2S) -2- (aminomethyl) -4-fluoropyrrolidine-1-carboxylic acid ester, 250mL of acetonitrile was added, and further 14.8g (0.11mol) of potassium carbonate was added thereto, and 10.2g (64.3mmol) of 1, 4-difluoro-2-nitrobenzene was added thereto. Under the protection of nitrogen, the temperature is raised to 80 ℃, the mixture is stirred for 1 hour, the TLC detects that the reaction is complete, water is added for 200mL for quenching, and ethyl acetate (200mL multiplied by 3) is extracted. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:40) to give benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-nitro-anilino) methyl ] -4-fluoropyrrolidine-1-carboxylic acid ester (18.1g, 86%).

Example 72: preparation of benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] -4-fluoropyrrolidine-1-carboxylic acid ester

Carbobenzoxy (2S) -2- [ (4-fluoro-2-nitro-anilino) methyl ] -4-fluoropyrrolidine-1-carboxylate 18.1g (46.3mmol) was taken in a 500mL reaction flask, 200mL acetic acid was added, 25g of iron powder was added thereto, stirring was carried out at room temperature for 16 hours, TLC detected for reaction completion, filtration was carried out, the filter cake was rinsed with ethyl acetate, the filtrate was concentrated under reduced pressure, 300mL water was added, ethyl acetate (200mL × 3) was extracted, the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. This gave benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] -4-fluoropyrrolidine-1-carboxylic acid ester (15.9g, 95%) which was used directly in the next reaction.

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

Taking 15.9g (44.0mmol) of carbobenzoxy (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] -4-fluoropyrrolidine-1-carboxylic ester in a 500mL reaction bottle, adding 300mL of 4N hydrochloric acid, adding 10g (0.132mol) of acetic alcohol, heating to 100 ℃, stirring for 6 hours, the reaction was detected by TLC to completion, cooled to 0 ℃, saturated sodium bicarbonate solution was added dropwise to adjust pH 8-9, filtered, and the filter cake was washed with methyl tert-butyl ether and dried to give (2S) -benzyl 4-fluoro-2- ((5-fluoro-2- (hydroxymethyl) -1H-benzo [ d ] imidazol-1-yl) methyl) pyrrolidine-1-carboxylate (15.7g, 89%) which was used directly in the next reaction.

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

Taking (2S) -benzyl 4-fluoro-2- ((5-fluoro-2- (hydroxymethyl) -1H-benzo [ d)]Imidazole-1-yl) methyl) pyrrolidine-1-carboxylate 15.7g (39.2mmol) is put into a 500mL reaction bottle, 300mL dichloromethane is added, under the protection of nitrogen, the temperature is reduced to 0 ℃, and then 20g (47.0mmol) of rumantin reagent is added, the temperature is restored to room temperature, the mixture is stirred for 2 hours, the TLC detection reaction is completed, the temperature is reduced to 0 ℃, and NaHCO is used for detecting the completion of the reaction₃/Na₂S₂O₃(1:1) the saturated solution (200mL x 2) was washed, the aqueous phase was extracted with dichloromethane (200mL x 2), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:5) to give (2S) -benzyl 4-fluoro-2- ((5-fluoro-2-formyl-1H-benzo [ d)]Imidazol-1-yl) methyl) pyrrolidine-1-carboxylic acid ester (14.4g, 92%).

Example 75: preparation of (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)

Taking 14.4g (36.1mmol) of (2S) -benzyl 4-fluoro-2- ((5-fluoro-2-formyl-1H-benzo [ d ] imidazol-1-yl) methyl) pyrrolidine-1-carboxylate, 15.6g (43.3mmol) of benzyloxycarbonyl (2S) -2- [ (4-fluoro-2-amino-anilino) methyl ] -4-fluoropyrrolidine-1-carboxylate in a 250mL reaction bottle, adding 150mLN, N-dimethylformamide, adding 27.4g (0.14mol) of sodium metabisulfite, stirring under nitrogen protection and microwave conditions for 1 hour under reflux, detecting complete TLC reaction, returning to room temperature, adding 500mL of water, extracting with ethyl acetate (200 mL. times.3), washing the combined organic layers with brine, drying with anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:5) to give (5S,5'S) -dibenzyl 5,5' - ((5,5 '-difluoro-1H, 1' H- [2,2 '-dibenzylo [ d ] imidazole ] -1,1' -diyl) bis (methylene)) bis (3-fluoropyrrolidine-1-carboxylate) (22.2g, 83%).

Example 76: preparation of 5,5 '-difluoro-1, 1' -bis ((2S) -4-fluoropyrrolidin-2-yl) methyl) -1H,1 'H-2, 2' -biizoyl [ d ] imidazole

22.2g (30.0mmol) of (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) was taken in a 250mL reaction flask, adding 100mL of methanol, adding 2g of palladium-carbon, stirring at room temperature for 12 hours under hydrogen atmosphere, TLC detected the reaction was complete, filtered, and the filtrate was concentrated to give 5,5 '-difluoro-1, 1' -bis ((2S) -4-fluoropyrrolidin-2-yl) methyl) -1H,1 'H-2, 2' -biimidazole (13.6g, 95% crude) which was used directly in the next reaction.

Example 77: preparation of 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-oxobutane-2, 1-diyl)) dicarbamate

In a 100mL reaction flask was taken 2.2g (10.2mmol) of N-Boc-L valine and 20mL of N-dimethylformamide was added, and in addition 2.1g (21.2mmol) of N-methylmorpholine and 12.7mmol of HATU4.8g (12.7mmol) were added thereto, and 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' -iodol [ d ] imidazole 2g (4.2mmol) was dissolved in 20mL of LN, N-dimethylformamide and added dropwise to the above system, stirred at room temperature for 3 hours, TLC detection of reaction was complete, 200mL of water was added, ethyl acetate (200 mL. times.3) was extracted, and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (eluent ethyl acetate: petroleum ether ═ 1:3) to give 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-oxobutane-2, 1-diyl)) dicarbamate (2.2g, 60%).

Example 78: preparation of (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)

To a 100mL reaction flask, 1.0g (1.15mmol) of 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-oxobutane-2, 1-diyl)) dicarbamate was added 20mL of an ethanol hydrochloride solution, and the mixture was stirred at room temperature for 1 hour to complete the reaction by TLC, and the solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 0.79g of a colorless oily liquid which was used directly in the next reaction.

Example 79: preparation of 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 (azepindiyl)) bis (1-oxopropane-2, 1-diyl)) bis (methyl carbamate)

0.58g (2.88mmol) of N-Boc-N-methyl-L-alanine was taken in a 100mL reaction flask, 20mL of N-dimethylformamide was added thereto, 0.61g (6.0mmol) of N-methylmorpholine and 0.37g (3.6mmol) of HATU1 were added thereto, and the mixture was stirred at room temperature for 30 minutes, and then 0.79g (crude, 1.15mmol) of (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-methylbutane-1-one) was taken and dissolved in 20mL of N-dimethylformamide, the mixture was added dropwise to the above system, stirred at room temperature for 3 hours, TLC checked for completion of the reaction, 200mL of water was added, ethyl acetate (200 mL. times.3) was extracted, and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by a silica gel column (eluent was ethyl acetate: petroleum ether ═ 1:3) to give 0.57g (48%) of 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-oxobutane-2, 1-diyl)) bis (azepindiyl)) bis (methyl 1-oxopropane-2, 1-diyl)) bis (methyl carbamate).

Example 80: preparation of (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)) bis (3-methyl-1-oxobutane-2, 1-diyl)) bis (2- (methylamino) propionamide)

Taking 0.57g (0.55mmol) of 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 (azepindiyl)) bis (1-oxopropane-2, 1-diyl)) bis (methyl carbamate) in a 100mL reaction flask, adding 20mL of hydrochloric acid ethanol solution, stirring at room temperature for 1 hour, detecting the reaction by TLC, distilling out the solvent and excess hydrogen chloride under reduced pressure, 100mL of water was added, pH was adjusted to 8 with saturated sodium bicarbonate, extracted with ethyl acetate (100mL × 2), and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by a silica gel column (eluent was ethyl acetate: petroleum ether ═ 1:2) to give 269mg (58%) of (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)) bis (3-methyl-1-oxobutane-2, 1-diyl)) bis (2- (methylamino) propionamide).

¹H-NMR(400MHz,d₆-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

The following compounds of examples were synthesized according to the synthesis methods of examples 69 to 80 and the first synthesis scheme:

example 81:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400 MHz,d₆-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:

¹H-NMR(400 MHz,d₆-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:

¹H-NMR(400 MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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

example 104:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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:

¹H-NMR(400MHz,d₆-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: binding affinity assay for Compounds to XIAP, cIAP1, cIAP2

Bir3 domain (10nM) was incubated with Smac polypeptide (10nM) in test buffer (50mM Tris, 120mM NaCl, 0.1% BSA, 1mM DTT, 0.05% TritonX100) in the presence of test compound for 1h at room temperature. The mixture was transferred to streptavidin-coated plates and incubated for 1h at room temperature to allow biotin-linked peptides to bind to the plates with the Bir3 structural domain. After several washes, Eu-labeled anti-GST antibody (Perkin Elmer DELFIA Eu-N1-anti GST, # AD0250) was added to each well (1: 5000 dilution with Perkin Elmer DELFIA test buffer 2013-01) and incubated at room temperature for 1 h. Washing with DELFIA washing buffer (Perkin Elmer DELFIA Wash2013-05) for 3 times, adding enhancement solution (Perkin Elmer enhancement solution 2013-02), incubating for 10min, detecting time-resolved fluorescence europium intensity, and calculating IC inhibition by compound with GraphPad Prizm5.0 software₅₀The value is obtained. The specific test results are shown in tables 1 and 2 below. Note: (A:<10nM B：10-100nM C：>100nM)

TABLE 1 binding affinity test results for the compounds of the invention to XIAP, cIAP1, cIAP2

TABLE 2 binding affinity test results for the inventive compounds and control drugs to XIAP, cIAP1, cIAP2

The experimental results show that the compounds of the invention have better binding affinity with XIAP, cIAP1 and cIAP2 proteins and better IAP inhibition activity, and the comparison results in table 1 further show that the binding affinity of some compounds of the invention for XIAP, cIAP1 and cIAP2 proteins is obviously better than that of control drugs.

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

Cells in the logarithmic growth phase were collected first. Counting, resuspending the cells in complete medium, adjusting the cell concentration to the appropriate concentration (as determined by cell density optimization assay) and plating 96-well plates with 100. mu.l cell suspension per well. Place the cells at 37 ℃ and 100% relative humidity, 5% CO₂Incubate in incubator for 24 hours. The compounds of the examples to be tested were diluted to the corresponding concentrations with the culture medium and the cells were added at 25. mu.l/well. The final concentration of compound was started at 100. mu.M and diluted in 4-fold gradient for 10 concentration points. Place the cells at 37 ℃ and 100% relative humidity, 5% CO₂Incubate in incubator for 72 hours. 10. mu.l of CCK-8 was added directly to the cell culture medium and incubated in an incubator at 37 ℃ for 2-4 hours. After gentle shaking, absorbance was measured on a SpectraMax M5Microplate Reader, and the inhibition rate was calculated. The specific test results are shown in tables 3 and 4 below.

Note: (A: <10nM B: 10-100nM C: 100-

TABLE 3 inhibition of MDA-MB-231 and PC-3 cell lines by the compounds of the invention

TABLE 4 results of inhibition of MDA-MB-231 and PC-3 cell lines by the compounds of the invention and the control drugs

The compound has better inhibition effect on cell growth in MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines. As can be seen from Table 4, the inhibitory effect of some compounds on MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines was significantly better than that of the control drug.

The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement 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 of formula I, and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof:

wherein R is₁And R₂Each is independently selected from;

R₃、R₄and R₅Each independently selected from H or optionally substituted with 1,2 or 3R: (C)₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, C_6-20Aryl or 5-14 membered heteroaryl;

ring a and ring B are each independently selected from: c_6-20Aryl or 5-14 membered heteroaryl;

R₆and R₇Each independently selected from halogen, hydroxy or from optionally substituted by 1,2 or 3R: (C)₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, C_6-20Aryl or 5-14 membered heteroaryl;

m and n are each independently selected from: 0.1, 2 or 3;

r is selected from halogen, CN, OH, SH, NH₂COOH, or selected from optionally substituted with 1,2 or 3R': (C)₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, C_6-20Aryl or 5-14 membered heteroaryl;

r' is selected from halogen, CN, OH, SH, NH₂、COOH；P₁And P₂Each independently selected from halogen, OH, CN, NH₂、COOH、(C₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) An aliphatic hydrocarbon group.

W is selected from a single bond, -O-, -S-, -NH-or optionally substituted with 1,2 or 3R: (C)₁-C₁₂) Aliphatic hydrocarbon radical, optionally containing one, two or more heteroatoms (C)₁-C₁₂) Aliphatic hydrocarbon radical, C_3-12Cycloalkyl, 3-12 membered heterocyclyl, C_6-20Aryl or 5-A 14-membered heteroaryl group.

2. A compound of formula I as claimed in claim 1, wherein the "optionally containing one, two or more heteroatoms of (C) optionally containing one, two or more heteroatoms, and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof₁-C₁₂) Aliphatic hydrocarbon groups ", the heteroatoms may be selected from sulphur, nitrogen, oxygen, phosphorus and silicon, optionally the heteroatoms are inserted in the aliphatic hydrocarbon groups, optionally C-C bonds and C-H bonds; for example, may be selected from (C)₁-C₁₂) Aliphatic hydrocarbyloxy, (C)₁-C₁₂) Aliphatic hydrocarbyl mercapto group, (C)₁-C₆) Aliphatic hydrocarbyloxy group (C)₁-C₆) Aliphatic hydrocarbyl mercapto group, (C)₁-C₆) Aliphatic hydrocarbyloxy (C)₁-C₆) Aliphatic hydrocarbon group, (C)₁-C₆) Aliphatic hydrocarbyl mercapto group (C)₁-C₆) Aliphatic hydrocarbon group, N- (C)₁-C₃) Aliphatic hydrocarbyl amino group (C)₁-C₆) Aliphatic hydrocarbon group, N-di- (C)₁-C₃) Aliphatic hydrocarbyl amino group (C)₁-C₆) An aliphatic hydrocarbon group; the halogen is selected from F, Cl, Br and I.

3. The compound of formula I as claimed in claim 1, wherein R is selected from the group consisting of racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs and pharmaceutically acceptable salts thereof, wherein R is selected from the group consisting of₃、R₄And R₅Can be selected from the following groups independently: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, ethenyl, 1-propenyl, 2-propenyl, 1-methylethenyl, 1-butenyl, 1-ethylethenyl, 1-methyl-2-propenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 2-methyl-2-propenyl, 1-pentenyl, 1-hexenyl, ethynyl, 1-propynyl, 2-propylpropenylAlkynyl, 1-butynyl, 1-methyl-2-propynyl, 3-butynyl, 1-pentynyl, 1-hexynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,

4. The compound of formula I as claimed in claim 1, wherein the structure of formula I is further selected from formula II, formula III, formula IV, formula V as follows:

in the formulas II, III, IV and V, R₁、R₂、R₆、R₇、P₁、P₂W, n, m, ring A, ring B are as defined for formula I.

5. The compound of formula I as claimed in claim 1, wherein the compound of formula I is selected from the group consisting of racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs thereof, or pharmaceutically acceptable salts thereof:

6. the process according to any one of claims 1 to 5 for the preparation of a compound of formula I, and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, wherein said process comprises the steps of:

wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、P₁、P₂W is as defined for formula I; r₃' selected from R₃Groups within the definition and independently selected from R₃Different groups; PG (Picture experts group)₁、PG₂Each represents a different amino protecting group selected from the group consisting of t-butyloxycarbonyl (N-boc), benzyloxycarbonyl protecting group (N-cbz), fluorenylmethoxycarbonyl protecting group (N-Fmoc), preferably, PG₁Selected from t-butyloxycarbonyl, PG₂Selected from benzyloxycarbonyl protecting groups.

7. The process according to any one of claims 1 to 5 for the preparation of a compound of formula I, and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, wherein said process comprises the steps of:

wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、P₁、P₂W is as defined for formula I; r₃' selected from R₃Groups within the definition and independently selected from R₃Different groups; PG (Picture experts group)₁、PG₂Each represents a different amino protecting group selected from the group consisting of t-butyloxycarbonyl (N-boc), benzyloxycarbonyl protecting group (N-cbz), fluorenylmethoxycarbonyl protecting group (N-Fmoc), preferably, PG₁Selected from t-butyloxycarbonyl, PG₂Selected from benzyloxycarbonyl protecting groups.

8. A pharmaceutical composition comprising a compound of formula I according to any one of claims 1-5 and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof.

9. Use of a compound of formula I according to any one of claims 1 to 5, and racemates, stereoisomers, tautomers, isotopic labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, prodrugs thereof or pharmaceutically acceptable salts thereof, for the preparation of an inhibitor of apoptosis proteins.

10. Use of a compound of formula I according to any one of claims 1-5, and racemates, stereoisomers, tautomers, isotopic labels, nitric oxides, solvates, polymorphs, metabolites, esters, prodrugs or pharmaceutically acceptable salts thereof, in the manufacture of a medicament for the treatment of a disease or condition due to an IAP disorder selected from a variety of benign or malignant tumors (cancers), 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 (peripheral neuroma), bladder cancer, glioblastoma, melanoma, adrenocortical cancer, AIDS-related lymphoma, anal cancer, bladder cancer, meningioma, glioma, astrocytoma, breast cancer, cervical cancer, chronic myeloproliferative 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 bile duct cancer, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gestational trophoblastoma, hairy cell leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hypopharynx cancer, intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, laryngeal carcinoma, leukemia, neuroblastoma, melanoma, neuroblastoma, melanoma, bladder cancer, colon cancer, melanoma, Acute lymphoblastic leukemia, acute myeloid leukemia, lip cancer, oral cancer, liver cancer, male breast cancer, malignant mesothelioma, medulloblastoma, melanoma, Merkel (Merkel) cell carcinoma, metastatic squamous neck cancer, multiple myeloma and other plasma cell neoplasms, mycosis fungoides and Sezary syndrome (Sezary syndrome), myelodysplastic syndrome, nasopharyngeal cancer, neuroblastoma, non-small cell lung cancer, oropharyngeal cancer, bone cancer (including malignant fibrous histiocytoma of osteosarcoma and bone), epithelial ovarian cancer, germ cell tumor, low malignant potential tumor of ovary (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, neuroblastoma, melanoma, and other plasma cell neoplasms of the human body, and other tumors of the human body, Rhabdomyosarcoma, salivary gland carcinoma, skin cancer, small bowel cancer, soft tissue sarcoma, supratentorial primary neuroectodermal tumor, pinealoblastoma, testicular cancer, thymoma, thymus cancer, thyroid cancer, transitional cell carcinoma of renal pelvis and ureter, cancer of urethra, uterine cancer, vaginal cancer, vulval cancer, and Wilms's tumor and other childhood renal tumors.