WO2020228478A1 - Anti-tumor diazobicyclic apoptosis protein inhibitor - Google Patents

Abstract

The present invention provides a novel apoptosis protein inhibitor or a pharmaceutically acceptable salt thereof, an isomer thereof, a preparation method therefor and a pharmaceutical composition, wherein the definition of each group is as shown in the description. The present invention also provides use of the compound, the pharmaceutically acceptable salt thereof and the isomer thereof in the preparation of a medicament for diseases related to IAP proteins. The compound of the present invention has a better binding affinity to XIAP, cIAP1 and cIAP2 proteins, has a better inhibiting effect on cell growth of MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines, and has larger pharmaceutical value and a broad market prospect.

Description

An anti-tumor inhibitor of diazobicyclic apoptotic protein

This application claims the priority of the prior application filed with the State Intellectual Property Office of China on May 16, 2019. The patent application number is 201910408971.2, and the invention title is "an anti-tumor inhibitor of diazobicyclic apoptosis protein". The full text of this application is incorporated into this application by reference.

Technical field

The field belongs to the field of medicinal chemistry, and specifically relates to an inhibitor of apoptosis protein and a preparation method and application thereof.

Background technique

Apoptosis or programmed cell death is a genetically and biochemically regulated mechanism, which plays an important role in the development and homeostasis of invertebrates and vertebrates. Abnormal apoptosis that leads to premature cell death has been linked to a variety of developmental disorders. Defects in apoptosis that result in a lack of cell death have been linked to cancer and chronic viral infections.

Current cancer therapies, including chemotherapeutics, radiation and immunotherapy, indirectly induce apoptosis in cancer cells. Therefore, the inability of cancer cells to perform the apoptosis program due to defects in the normal apoptosis mechanism is usually associated with increased resistance to apoptosis induced by chemotherapy, radiation or immunotherapy. Such primary or acquired resistance of cancer to current therapies due to defects in apoptosis is a major problem in current cancer therapies. One type of central negative regulator of apoptosis is the inhibitor of apoptosis protein (IAP). This category includes proteins such as XIAP, cIAP1, cIAP2, ML-IAP, HIAP, KIAP, TSIAP, NAIP, survivin, livin, ILP-2, apollon and BRUCE. IAP protein effectively inhibits the apoptosis of cancer cells induced by many kinds of apoptosis stimuli (including chemotherapeutics, radiation and immunotherapy), and is a new target of the apoptosis pathway.

Patent applications for inventions involving IAP inhibitors include WO2011018474A1, WO2008016893A1, WO2014047024A1, CN101484151A, etc.

IAP inhibitors currently under development include LCL161, Birinapant, BV6, GDC-0152, AZD5582, AT406, etc. Among them, AT-406 is an effective, oral Smac mimic, an antagonist of IAPs, which can inhibit XIAP. The structural formulas of cIAP1 and cIAP2 proteins are as follows:

At present, the clinical use of immune checkpoint inhibitors alone has a weak effect on solid tumors. The combination of IAP inhibitors and immune checkpoint inhibitors can achieve better therapeutic effects in solid tumors. IAP inhibitors have a synergistic immune checkpoint inhibitor treatment The role of cancer has very good application prospects, but there are still many major challenges in the development of this species. In order to meet the current clinical needs for apoptosis protein inhibitors and achieve better treatment effects for tumor diseases, we are committed to the research and development of a series of high-efficiency and low-toxic IAP inhibitors, which are of great importance to the medical field. significance.

Summary of the invention

The purpose of the present invention is to provide a new inhibitor of apoptosis protein or a pharmaceutically acceptable salt or isomer thereof, characterized in that the compound has the following general formula:

Wherein, X represents a C or N atom, when X is a C atom, Y is a CONHR ₃ and Z represents a H atom; when X is a N atom, Z is a CONHR ₃ and Y does not exist;

The R ₁ and R ₂ are each independently selected from C _1-20 alkylamino groups, C _5-8 cycloalkylamino groups,

C _1～20 alkyl group,

Or α-amino acid residues,

The α-amino acid residue is a substituent formed by the deletion of the carboxyl group on the α-carbon in the α-amino acid; the Ar group represents an aryl or heteroaryl group with 5-8 atoms, The m and n are each independently selected from any integer of 1 to 3;

R _{3 is} selected from substituted or unsubstituted benzyl, and the substituent is optionally selected from phenyl, halogen, C _1-4 alkyl or p-fluorophenyl;

Further, a compound or a pharmaceutically acceptable salt or isomer thereof, the compound has the following general formula:

or

Wherein, in the above formula I or II,

Said R _{1 is} selected from C _1-20 alkylamino groups,

C _1～20 alkyl group,

Or any of the α-amino acid residues,

R _{2 is} selected from C _1-20 alkylamino group, C _5-8 cycloalkylamino group, C _1-20 alkyl group,

Or any of the α-amino acid residues,

The α-amino acid residue is a substituted residue formed by the deletion of the carboxyl group on the α-carbon in the α-amino acid, and the Ar group represents an aryl group and a heteroaryl group with 5-8 atoms. , The m and n are each independently selected from any integer of 1 to 3,

R _{3 is} selected from substituted or unsubstituted benzyl, and the substituent is optionally selected from phenyl, halogen, C _1-4 alkyl or p-fluorophenyl;

In one approach, the alkylamino group described in formula I or II is

And its isomers;

In one scheme, the C _5-6 cycloalkylamino group described in formula I or II is

In one scheme, the α-amino acid residue is

or

In one scheme, R ₂ in formula I or II is selected from C _1-13 alkyl;

In one approach, the aryl group described in formula I or II is phenyl;

In one approach, the heteroaryl group described in formula I or II is imidazolyl;

Preferably, in the R ₃ group, the halogen is F.

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

The present invention also provides methods for preparing compounds of formula I and formula II and their salts or isomers, but are not limited to the methods described below. All raw materials are prepared according to the group characteristics of the target molecule in accordance with the general formula, and are prepared through the schemes in these routes and methods well known to those of ordinary skill in the organic chemistry field or directly purchased. The compounds of the present invention can be synthesized by combining the following methods with synthetic methods known in the field of synthetic organic chemistry or related modification methods recognized by those skilled in the art.

In one scheme, the preparation route of the compound of the present invention is as follows:

plan 1

Option 1 includes the following steps:

Take N-Boc pyrrolidine-2-carboxylic acid methyl ester as starting material, react with allyl bromide under the action of lithium bistrimethylsilylamide, etc. to produce compound 1. The reaction temperature is selected from -80℃ to -20℃ , The reaction solvent is selected from inert aprotic solvents such as tetrahydrofuran and toluene;

The Boc protecting group of compound 1 is removed under acidic conditions to obtain compound 2. The acid can be hydrochloric acid, p-toluenesulfonic acid, etc., and the reaction solvent can be methanol, ethanol, tetrahydrofuran, etc.;

Compound 2 under the action of condensing agent, and

The reaction produces compound 3, P ₁ and P ₂ are protecting groups for amino groups, and you can choose tert-butoxycarbonyl (Boc), 9-fluorenylmethylenoxycarbonyl (Fmoc), Boxycarbonyl (Cbz), and allyloxycarbonyl (Alloc), triphenylmethyl (Trt) and any suitable protecting groups, P _{1 is} not equal to P ₂ (in embodiments, P ₁ and P ₂ are orthogonal protecting groups). The condensing agent can be selected from N,N'-diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2-(7- Nitrobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and other condensing agents;

Under ozone conditions, the carbon-carbon double bond is oxidized to an aldehyde with one less carbon to obtain compound 4. The reaction temperature is from -80℃ to -20℃, the reaction solvent is tetrahydrofuran, dichloromethane, methanol, and the reducing agent is selected Triphenylphosphine, sodium sulfite, triethylamine, etc.;

Compound 4 removes the protective group P ₂ under acidic conditions (when P ₂ is the Boc protecting group), palladium-carbon hydrogenation (when P ₂ is the Cbz protecting group) or piperidine (when P ₂ is the Fmoc protecting group), and further Reductive amination to obtain macrocyclic compound 5, the reaction solvent is tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the reducing agent is sodium borohydride, sodium cyanoborohydride, etc.;

Compound 5 is in an inert organic solvent such as acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, under the action of a condensing agent, and a carboxylic acid compound containing R ₁ group undergoes condensation reaction to obtain compound 6, temperature Choose from 25℃ to 120℃, choose N,N'-diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, condensing agent 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluoro Phosphate etc.;

Compound 6 is saponified under alkaline conditions, and then the pH value is controlled under acidic conditions to obtain compound 7. The organic solvent is methanol, ethanol and other water-soluble solvents, and the alkali is inorganic alkalis such as lithium hydroxide and sodium hydroxide;

Compound 7 is in inert organic solvent such as acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, under the action of a condensing agent, and an amino compound containing R ₃ group is condensed to obtain compound 8, and the temperature is selected At 25°C to 120°C, the condensing agent is N,N'-diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2 -(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate Salt etc.;

Compound 8 removes the protective group P ₁ under acidic conditions (when P ₁ is a Boc protecting group), palladium-carbon hydrogenation (when P ₁ is a Cbz protecting group), or piperidine (when P ₁ is a Fmoc protecting group), etc. Amino compound 9, the reaction solvent is tetrahydrofuran, dichloromethane, methanol, toluene, etc.;

Compound 9 is in an inert organic solvent such as acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, under the action of a condensing agent, and a carboxylic acid compound containing R ₂ group undergoes a condensation reaction to obtain compound I. Choose from 25℃ to 120℃, choose N,N'-diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, condensing agent 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluoro Phosphate etc.

In another scheme, the preparation route of the compound of the present invention is as follows:

Scenario 2

Option 2 includes the following steps:

Using compound 10 as the starting material, the ring is closed under the action of inorganic bases such as potassium carbonate and cesium carbonate to produce compound 11. The reaction temperature is 25°C to 120°C, and the reaction solvent is acetonitrile, tetrahydrofuran, and N,N-dimethylformaldehyde. Inert aprotic solvents such as amides;

Compound 11 under the action of condensing agent, and

The reaction produces compound 12, P ₁ and P ₂ are protecting groups for amino groups, and you can choose tert-butoxycarbonyl (Boc), 9-fluorenylmethylenoxycarbonyl (Fmoc), Boxycarbonyl (Cbz), and allyloxycarbonyl (Alloc), triphenylmethyl (Trt), etc., P _{1 is} not equal to P ₂ (in embodiments, P ₁ and P ₂ are orthogonal protecting groups). Condensing agent selected N,N'-diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2-(7-azobenzene Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, etc.;

Under ozone conditions, the carbon-carbon double bond of compound 12 is oxidized to an aldehyde with one less carbon to obtain compound 13. The reaction temperature is selected from -80°C to -20°C, the reaction solvent is tetrahydrofuran, dichloromethane, methanol, and the reducing agent is selected Triphenylphosphine, sodium sulfite, triethylamine, etc.;

Compound 13 removes the protective group P ₂ under acidic conditions (when P ₂ is the Boc protecting group), palladium-carbon hydrogenation (when P ₂ is the Cbz protecting group), or piperidine (when P ₂ is the Fmoc protecting group), and further Reductive amination to obtain macrocyclic compound 14, the reaction solvent is tetrahydrofuran, dichloromethane, methanol, toluene, etc., and the reducing agent is sodium borohydride, sodium cyanoborohydride, etc.;

Compound 14 is in inert organic solvent such as acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, under the action of a condensing agent, and a carboxylic acid compound containing R ₁ group undergoes condensation reaction to obtain compound 15. Choose from 25℃ to 120℃, choose N,N'-diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, condensing agent 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluoro Phosphate etc.;

Compound 15 is saponified under alkaline conditions, and then the pH value is controlled under acidic conditions to obtain compound 16. The organic solvent is methanol, ethanol and other water-soluble solvents, and the alkali is inorganic alkalis such as lithium hydroxide and sodium hydroxide;

Compound 16 is in inert organic solvents such as acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, under the action of a condensing agent, and an amino compound containing R ₃ group undergoes condensation reaction to obtain compound 17, and the temperature is selected At 25°C to 120°C, the condensing agent is N,N'-diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2 -(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate Salt etc.;

Compound 17 removes the protective group P ₁ under acidic conditions (when P ₁ is a Boc protecting group), palladium-carbon hydrogenation (when P ₁ is a Cbz protecting group), or piperidine (when P ₁ is a Fmoc protecting group), etc. Amino compound 18, the reaction solvent is tetrahydrofuran, dichloromethane, methanol, toluene, etc.;

Compound 18 is in an inert organic solvent such as acetonitrile, toluene, chloroform, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, under the action of a condensing agent, and a carboxylic acid compound containing R ₂ group undergoes condensation reaction to obtain compound II, temperature Choose from 25℃ to 120℃, choose N,N'-diisopropylcarbodiimide, benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, condensing agent 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluoro Phosphate etc.

Further, in the above scheme 1 and scheme 2, the P ₁ and P ₂ groups are the protecting groups of the amino group, and the R ₁ , R ₂ and R ₃ groups are defined as described above in the specification.

The preparation of the compound of formula I'can refer to the method of Scheme 1 or Scheme 2.

In the above preparation methods, the method for preparing compounds involves protecting and deprotecting various chemical groups, and those skilled in the art can easily select a suitable protecting group according to the needs of protection and deprotection. The above-mentioned group features mentioned in the present invention or the features mentioned in the examples can be combined arbitrarily on the basis of compliance with the laws of pharmacy. Each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equal or similar purpose. . Unless otherwise specified, the disclosed features are only general examples of the same or similar features.

Another aspect of the present invention provides a pharmaceutical composition, which contains a therapeutically effective amount of any compound selected from the compound of formula I'or formula I or formula II or a pharmaceutically acceptable salt or isomer thereof as an active ingredient , And one or more pharmaceutically acceptable carriers, diluents and/or excipients.

The pharmaceutical composition preferably contains a compound of formula I'or formula I or formula II or a pharmaceutically acceptable salt or isomer thereof in a weight ratio of 1% to 90% as an active ingredient, more preferably a weight ratio of 10 %-80% active ingredients.

Unless otherwise stated, the following terms used in the claims and specification have the following meanings or features:

The term "C _{1 ~ 20} alkyl group" means a saturated aliphatic hydrocarbon group of 1 to 20 carbon atoms, including straight-chain and branched-chain groups (the numerical ranges mentioned in this application, for example, "1-20", refers to The group, in this case an alkyl group, can contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to 20 carbon atoms). The alkyl group in the present invention includes "alkylene". An alkyl group containing 1 to 6 carbon atoms is called a lower alkyl group. When a lower alkyl group has no substituent, it is called an unsubstituted lower alkyl group. More preferably, the alkyl group is a medium-sized alkyl group having 1-10 carbon atoms, such as methyl, ethyl, ethylene, propyl, propylene, 2-propyl, n-butyl, isopropyl Butyl, butylene, tert-butyl, pentyl, etc. Preferably, the alkyl group is a lower alkyl group having 1 to 5 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, butylene, isobutyl or tert-butyl. Alkyl groups can be substituted or unsubstituted.

The term "C _1-20 alkylamino group" means an R-amino compound, and the definition of R is as described for the C _1-20 alkyl group.

The term "C _5-8 cycloalkaneamino group" means an amine compound formed by substituting one carbon on a C _5-8 cycloalkane with an amino group, for example

Wait.

The term "α-amino acid residue" means a substitution residue formed after the carboxyl group on the α-carbon of an amino acid is deleted.

The term "benzyl" means "benzyl", with a simple structure of C ₆ H ₅ CH ₂ -, or Bn (abbreviation for Benzyl).

The term "aryl" refers to an all-carbon monocyclic or fused polycyclic group of 5 to 12 carbon atoms with a fully conjugated π-electron system. Non-limiting examples of aryl groups are phenyl, naphthyl and anthracenyl. The aryl ring may be fused to a heteroaryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring. Aryl groups can be substituted or unsubstituted. When substituted, the substituent is preferably one or more, more preferably one, two or three, and even more preferably one or two, independently selected from lower alkyl, trihaloalkyl, halogen, and hydroxyl , Lower alkoxy, mercapto, (lower alkyl)thio, cyano, acyl, thioacyl, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thioamino Formyl, C-amido, N-amido, nitro, N-sulfonamido, S-sulfonamido. Preferably, the aryl group is a 5-membered monocyclic aryl group or a 6-membered monocyclic aryl group.

The term "heteroaryl" means a monocyclic or condensed ring group of 5 to 12 ring atoms, containing one, two, three or four ring heteroatoms selected from N, O or S, and the remaining ring atoms are C, in addition has a fully conjugated π electron system. The heteroaryl ring may be fused to an aryl, heterocyclic or cycloalkyl ring, and the ring connected to the parent structure is a heteroaryl ring. Heteroaryl groups can be substituted or unsubstituted. When substituted, the substituent is preferably one or more, more preferably one, two or three, and still more preferably one or two. Non-limiting examples of unsubstituted heteroaromatic bases are pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyrimidine, quinoline, isoquinoline, purine, tetrazole, triazine and carbazole; preferably, Heteroaryl groups are nitrogen-containing 5-membered monocyclic heteroaryl groups and nitrogen-containing 6-membered monocyclic heteroaryl groups.

The term "isomer" can be understood to cover the conventional compound "stereoisomerism" in the art, including enantiomers and diastereomers, where diastereomers include cis and trans isomers, for example, From its cis isomer, trans isomer or a mixture of cis and trans isomers.

The term "halogen" means fluorine, chlorine, bromine or iodine.

The term "pharmaceutically acceptable salts" means those salts that retain the biological effectiveness and properties of the parent compound. Such salts include:

(1) Forming a salt with an acid, obtained by the reaction of the free base of the parent compound with an inorganic acid or organic acid, such as (but not limited to) hydrochloric acid, hydrobromic acid, etc., and organic acid such as (but not limited to) acetic acid, Malic acid, fumaric acid, maleic acid, p-toluenesulfonic acid, tartaric acid, citric acid, lactic acid, succinic acid or malonic acid, etc. Such salts have safety, effectiveness and due biological activity when used in mammals.

"Pharmaceutical composition" refers to one or more of the compounds described herein or their pharmaceutically acceptable salts, isomers and prodrugs and other chemical components, such as pharmaceutically acceptable carriers and Mixture of excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.

"Pharmaceutically acceptable carrier" refers to a carrier or diluent that does not cause significant irritation to organisms and does not interfere with the biological activity and properties of the administered compound.

"Excipient" refers to an inert substance added to a pharmaceutical composition to further facilitate the administration of a compound. Examples of excipients include (not limited to) lactose, sucrose, microcrystalline cellulose, sorbitol, polyvinylpyrrolidone, cellulose, water, methylcellulose, and the like.

The pharmaceutical composition may also contain: lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives such as methyl benzoate, etc.; sweeteners and flavoring agents.

The composition of the present invention is formulated, dosed and administered in a manner consistent with good medical practice. The factors to be considered in this situation include the specific condition being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the condition, the location where the agent is delivered, the method of administration, the dosing regimen, and what is known to the medical practitioner Other factors.

The present invention also provides the use of the compound of formula I'or formula I or formula II or a pharmaceutically acceptable salt or isomer thereof:

(1) It is used to induce apoptosis in cells, or make cells, especially cancer cells, sensitive to apoptosis signals.

(2) It is used to induce apoptosis in cells overexpressing IAP protein (for example, c-IAP1, c-IAP2, X-IAP or ML-IAP).

(3) Preparation of drugs for diseases related to IAP protein; more broadly, the compounds can be used to treat cancer.

(4) Further, the compounds of the present invention can be used to treat all types of cancer drugs that fail to undergo apoptosis. Examples of such cancer types include neuroblastoma, bowel cancer such as rectal cancer, colon cancer, familial adenomatous polyp cancer and hereditary non-polyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, Tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine body cancer, endometrial cancer, choriocarcinoma, pancreatic cancer , Prostate cancer, testicular cancer, breast cancer, urinary system cancer, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal tumor, Hodgkin's lymph Tumor, non-Hodgkin’s lymphoma, Burkitt’s lymphoma, acute lymphatic leukemia (ALL), chronic lymphatic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T- Cell leukemia lymphoma, hepatocellular carcinoma, gallbladder cancer, bronchial cancer, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, spermatogonia Tumor, rhabdomyosarcoma, craniopharyngeoma, osteosarcoma, chondrosarcoma, sarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma and plasmacytoma. Can be used to treat solid tumors. It can also be used to treat many major diseases such as breast cancer, pancreatic cancer or malignant melanoma.

The positive control compound AT-406 of the compound of the present invention is an inhibitor of cIAP1 and cIAP2, and its inhibitory effect on XIAP is weaker than that of cIAP1 and cIAP2, while the compound of the present invention has stronger inhibitory activity on the three proteins of XIAP, cIAP1 and cIAP2. Therefore, the compound of the present invention has a stronger inhibitory effect on apoptosis proteins as a whole;

From cell pharmacological data, it is found that the compound of the present invention has a strong inhibitory effect on MDA-MB-231 breast cancer and PC-3 pancreatic cancer cells, while the positive control compound AT-406 only has a better effect on MDA-MB-231 breast cancer. Strong, the inhibitory effect on PC-3 pancreatic cancer cells is relatively weak;

The compound of the present invention has good IAP inhibitory activity, and can be developed into a cancer treatment drug used alone in the later stage, or can be combined with other targeted drugs to treat cancer;

In summary, the compound of the present invention has good binding affinity with XIAP, cIAP1, and cIAP2 proteins, and has a good inhibitory effect on cell growth in MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines. Its medicinal value and broad market prospects.

Specific embodiment

The following specific examples are used to further illustrate the present invention, but the present invention is by no means limited to these examples (unless otherwise noted, all raw materials are commercially available).

(All the chiral compounds involved in this example are prepared and purified by chiral column)

Example 1: Preparation of methyl pyrrolidine-2-carboxylate

Take 40g (0.35mol) of proline and 250mL of methanol in a 500mL single-necked reaction flask, cool to 0℃, take another 45g (0.38mol) of thionyl chloride in a 250mL constant pressure dropping funnel, and slowly add dropwise to the above In the solution system, heat is released with the dropping system, and the internal temperature is controlled to be less than 5°C, and the dropping is completed in about 20 minutes. The temperature was raised to 70°C to obtain a clear solution, which was refluxed for 3 hours. After cooling to room temperature, the solvent was evaporated under reduced pressure to obtain 43 g of light yellow oily liquid, and the crude product was directly used in the next reaction.

Example 2: Preparation of Methyl N-Bocpyrrolidine-2-carboxylate

Take 43g (0.33mol) of methyl pyrrolidine-2-formate and 400mL of dichloromethane in a 2L single-necked reaction flask, add 88g (0.87mol) of triethylamine to obtain a white emulsion, stir at room temperature for 15 minutes, and cool to 0°C.

In addition, 152 g (0.69 mol) of Boc anhydride was dissolved in 600 mL of dichloromethane in a constant pressure funnel, and added dropwise to the above system. A large number of bubbles were released with the dropwise addition, heat was released, and the dropping rate was controlled to maintain the internal temperature of the system <10°C. After the addition was completed, a white suspension was obtained, which was stirred at room temperature for 12 hours. The white precipitate was removed by filtration, and the filtrate was evaporated under reduced pressure to obtain a white suspension, which was washed with 300 mL of citric acid solution (0.5M), extracted with ether (3*500 mL), combined the organic layers, washed with saturated brine, and used It was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was purified by column chromatography (eluent: ethyl acetate: petroleum ether = 1:5) to obtain 75 g of a clear and colorless oily liquid with a yield of 94%.

Example 3: Preparation of N-Boc-2-propenyl-pyrrolidine-2-carboxylic acid methyl ester

Take 20 g (87.3 mmol) of N-Boc pyrrolidine-2-carboxylic acid methyl ester and 200 mL of tetrahydrofuran in a 500 mL three-necked flask under nitrogen protection and cool to -78°C.

In addition, 105 mL (105 mmol, 1M) of lithium bistrimethylsilylamide was added to the constant pressure dropping funnel and added dropwise to the above system to control the dropping rate and maintain the internal temperature of the system <-78°C to obtain a yellow-brown solution. After the addition is complete, stir at -78°C for 1.5 hours.

In addition, 15.9 g (131 mmol) of allyl bromide was dissolved in 100 mL of tetrahydrofuran, and added dropwise to the above system, controlling the dropping rate, maintaining the internal temperature of the system <-78°C, after the addition, stirring at -78°C for 1 hour. Return to room temperature and stir for 1 hour to obtain a reddish brown solution. TLC detected that the reaction was complete, and 50 mL of water was added to quench the reaction. Ethyl acetate extraction (3*200 mL) and the combined organic layers were washed with saturated brine. The organic layer was dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was subjected to column chromatography (eluent: ethyl acetate: petroleum ether = 1:10) Purification to obtain 18.7 g of colorless oily liquid with a yield of 82%.

Example 4: Preparation of 2-propenyl-1-((3-benzyloxycarbonyl-amino-2-tert-butoxycarbonyl-amino)propionyl)pyrrolidine-2-carboxylic acid methyl ester

Take 18g (66.9mmol) of N-Boc-2-propenyl-pyrrolidine-2-carboxylic acid methyl ester and 50mL of absolute ethanol in a 500mL single-necked reaction flask, add 100mL of hydrochloric acid ethanol, stir at room temperature for 2 hours, TLC detection, the reaction is complete . The solvent and excess hydrogen chloride were evaporated under reduced pressure to obtain 14 g of a colorless oily liquid, which was dissolved in 50 mL of dichloromethane.

Separately take 27g (80.3mmol) of (S)-2-(Boc-amino)-3-(Cbz-amino)propionic acid and 200mL of dichloromethane in a 500mL single-neck reaction flask, add 38g HATU (100.35mmol), 17gDIEA( 133.8mmol), stirred at room temperature for 1 hour to obtain a yellow clear liquid.

The dichloromethane solution prepared in the first step was added dropwise to the yellow clear solution, and stirred at room temperature for 6 hours to obtain a reddish brown solution. TLC detected that the reaction was complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 400 mL ethyl acetate, washed with 1M HCl solution, saturated sodium bicarbonate solution, saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by column chromatography (eluent: ethyl acetate: petroleum ether = 1:2) to obtain 21.2 g of white solid with a yield of 65%.

Example 5: Preparation of 2-acetaldehyde-1-((3-benzyloxycarbonyl-amino-2-tert-butoxycarbonyl-amino)propionyl)pyrrolidine-2-carboxylic acid methyl ester

Take 20g (40.9mmol) of methyl 2-propenyl-1-((3-benzyloxycarbonyl-amino-2-tert-butoxycarbonyl-amino)propionyl)pyrrolidine-2-carboxylate and 200mL dichloromethane in 500mL In a three-necked reaction flask, cool to -78°C, and pass ozone for about 30 minutes until the system turns blue. Stir at -78°C for 30 minutes, stop passing ozone. Nitrogen was bubbled through to remove excess ozone until the blue color disappeared, and a colorless solution was obtained. 100 mL of triethylamine was added, returned to room temperature, and stirred for 2 hours. TLC detected that the reaction was complete. Wash with 1M citric acid, wash with saturated sodium chloride aqueous solution, dry the organic layer with sodium sulfate, filter, distill off the solvent under reduced pressure, and purify the residue by column chromatography (eluent: ethyl acetate: petroleum ether = 1:2) , To obtain 16.8 g of white solid with a yield of 84%.

Example 6: (5R,10aR)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrrolo[1,2-a][1,5]diazaoctane-10a- Preparation of methyl formate

Take 15g (30.5mmol) 2-acetaldehyde-1-((3-N-benzyloxycarbonyl-2-N-tert-butoxycarbonyl)propionyl)pyrrolidine-2-carboxylic acid methyl ester and 260mL methanol in 500mL single mouth In the reaction flask, it was dissolved into a colorless clear liquid, 1.5 g (10%) Pd/C was added, and the mixture was stirred at room temperature for 12 hours under a hydrogen atmosphere. TLC detected that the reaction was complete. The Pd/C was removed by filtration, the solvent was distilled off under reduced pressure, 200 mL of tetrahydrofuran was added to dissolve, 2.9 g (45.8 mmol) of sodium cyanoborohydride was added, and the mixture was stirred at room temperature. TLC detected that the reaction was complete. The reaction was quenched by adding 20 mL of water, filtered, and the solvent was evaporated under reduced pressure. The residue was dissolved in 200 mL of ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. After purification (eluent: dichloromethane:methanol=10:1), 7.8 g of white solid was obtained with a yield of 75%.

Example 7: (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrrole Preparation of and [1,2-a][1,5]diazaoctane-10a-methyl carboxylate

Take 1.5g (6.5mmol) of N-Cbz-N-methyl-D-alanine and 60mL of dichloromethane in a 250mL single-mouth reaction flask, add 3.4g HATU (8.9mmol), 1.5gDIEA (11.8mmol), room temperature Stir for 1 hour to obtain a yellow clear liquid.

Take (5R,10aR)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrrolo[1,2-a][1,5]diazaoctane-10a-carboxylic acid methyl ester 2 g (5.9 mmol) was dissolved in 20 mL of dichloromethane, added dropwise to the above system, stirred at room temperature for 6 hours, TLC detected that the reaction was complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 200 mL ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent was dichloromethane : Methanol=20:1) Purification to obtain 2.1 g of brown oil with a yield of 64%.

Example 8: (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrrole Preparation of bi[1,2-a][1,5]diazaoctane-10a-carboxylic acid

Take 2g (3.6mmol) (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-de Hydropyrrolo[1,2-a][1,5]diazepine-10a-methyl carboxylate and 60mL methanol in a 250mL single-necked reaction flask, add 0.4g potassium hydroxide (7.2mmol), 60mL water, Heat to 50°C and stir for 3 hours. TLC detects that the reaction is complete. 60mL of water was added, methanol was evaporated under reduced pressure, the remaining aqueous solution was extracted with ethyl acetate (2*100mL), the organic layer was discarded, the aqueous solution was adjusted to pH 1-2 with concentrated hydrochloric acid, and extracted with ethyl acetate (3*100mL), organic The layer was dried with sodium sulfate, filtered, and the solvent was distilled off under reduced pressure to obtain 1.9 g of oil, which was directly used in the next reaction.

Example 9: (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrrole Preparation of and [1,2-a][1,5]diazaoctane-10a-(N-diphenylmethyl)-carboxamide

Take (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrrolo[1 ,2-a][1,5]Diazaoctane-10a-carboxylic acid 1.8g (3.3mmol), 2.6g PyBop (5.0mmol) in a 100mL single-mouth reaction flask, add 50mL DMF, and take another 0.85g DIEA( 6.6mmol), stirred at room temperature for 30 minutes. Another 1.2 g (6.6 mmol) of benzhydrylamine was added to the system, heated to 90° C., and stirred for 12 hours. TLC detected that the reaction was complete. The reaction solution was poured into 300 mL of water, extracted with ethyl acetate (3*100 mL), washed with saturated sodium chloride solution, the organic layer was dried with sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent is two Chloromethane: methanol = 15:1) was purified to obtain 1.7 g of light yellow solid with a yield of 74%.

Example 10: (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-amino-6-one-dehydropyrrolo[1,2-a] [1,5] Preparation of Diazaoctane-10a-(N-Diphenylmethyl)-Carboxamide

Take 0.5g (0.7mmol) (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one- Dehydropyrrolo[1,2-a][1,5]diazepine-10a-(N-diphenylmethyl)-carboxamide and 10mL absolute ethanol in a 100mL single-neck reaction flask, add 20mL hydrochloric acid Ethanol, stirring at room temperature for 2 hours, TLC detection, the reaction is complete. The solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 0.5 g of a colorless oily liquid, which was directly used in the next reaction.

Example 11: (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrolo[ Preparation of 1,2-a][1,5]diazaoctane-10a-(N-diphenylmethyl)-carboxamide

Take 82 mg (0.8 mmol) of isovaleric acid and 10 mL of dichloromethane in a 100 mL single-necked reaction flask, add 0.42 g HATU (1.1 mmol), 0.18 g DIEA (1.4 mmol), and stir at room temperature for 30 minutes to obtain a light yellow clear liquid.

Take (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-amino-6-one-dehydropyrrolo[1,2-a][1, 5] Diazaoctane-10a-(N-diphenylmethyl)-carboxamide 0.5g dissolved in 10mL dichloromethane, added to the above system to obtain a yellow-green solution, stirred at room temperature for 3 hours, TLC detection, reaction complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 100 mL ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent was dichloromethane : Methanol=20:1) Purification to obtain 370 mg of pale yellow solid with a yield of 76%.

Example 12: (5R,10aR)-3-((R)-2-methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrolo[1,2-a ][1,5]Diazaoctane-10a-(N-diphenylmethyl)-carboxamide)(I-11)

Take (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrolo[1,2 -a][1,5] Diazaoctane-10a-(N-Benzhydryl)-carboxamide 200mg (0.29mmol) and 50mL methanol in a 100mL single-necked reaction flask, dissolved into a colorless clear liquid, add 20mg (10%) Pd/C, stirred at room temperature for 12 hours under hydrogen atmosphere, TLC detected that the reaction was complete. The Pd/C was removed by filtration, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane:methanol=15:1) to obtain 138 mg of pale yellow solid with a yield of 85%.

¹ H NMR (400MHz, d ₆ -DMSO): δ=8.79-8.82 (m, 1H), 7.90-8.10 (m, 1H), 7.22-7.36 (m, 8H), 7.13-7.15 (m, 2H), 6.03-6.06(m,1H),4.75-4.90(m,1H),3.92-4.00(m,1H),3.65-3.80(m,1H),3.52(s, 1H),3.40-3.43(m,2H) ), 2.90-3.20 (m, 3H), 2.10-2.30 (m, 5H), 1.60-2.00 (m, 5H), 1.24 (s, 2H), 1.05-1.15 (m, 3H), 0.84-0.89 (m ,6H), [M+H] ⁺ :m/z=562.3

Example 13: (5R,10aR)-3-benzyloxycarbonyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrrolo[1,2-a][1,5]diazepine Preparation of Heterooctyl-10a-Methyl Formate

Take (5R,10aR)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrrolo[1,2-a][1,5]diazaoctane-10a-carboxylic acid methyl ester 5g (14.7mmol), 3.1g (36.8mmol) of sodium bicarbonate and 30mL of tetrahydrofuran were put into a 100mL single-necked reaction flask, 30mL of water was added, cooled to 5°C, another 3.0g (17.6mmol) of Cbz-Cl was dissolved in 15mL of tetrahydrofuran, Add dropwise to it, after the dropwise addition is complete, stirring at room temperature for 6 hours, TLC detects that the reaction is complete. After filtration, the filtrate was evaporated under reduced pressure to remove tetrahydrofuran, extracted with ethyl acetate (3*100 mL), washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatography (eluent (Dichloromethane:methanol=20:1) to obtain 6.4 g of white solid with a yield of 92%.

Example 14: (5R,10aR)-3-benzyloxycarbonyl-5-amino-6-one-dehydropyrrolo[1,2-a][1,5]diazaoctane-10a-formic acid methyl Preparation of ester

Take 6g (12.6mmol) (5R,10aR)-3-benzyloxycarbonyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrrolo[1,2-a][1,5] Diazaoctane-10a-methyl formate and 30 mL of absolute ethanol were put into a 100 mL single-necked reaction flask, 30 mL of hydrochloric acid ethanol was added, and the mixture was stirred at room temperature for 2 hours. TLC detected that the reaction was complete. The solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 5.2 g of a colorless oily liquid, which was directly used in the next reaction.

Example 15: (5R,10aR)-3-benzyloxycarbonyl-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrrolo[1 Preparation of ,2-a][1,5]diazaoctane-10a-methyl carboxylate

Take 2.8g (13.9mmol) of N-Boc-N-methyl-D-alanine and 60mL of dichloromethane in a 250mL single-mouth reaction flask, add 7.2g HATU (18.9mmol), 3.3g DIEA (25.2mmol), room temperature Stir for 1 hour to obtain a yellow clear liquid.

Take (5R,10aR)-3-benzyloxycarbonyl-5-amino-6-one-dehydropyrrolo[1,2-a][1,5]diazaoctane-10a-carboxylic acid methyl ester 5.2g Dissolved in 20 mL of dichloromethane, added dropwise to the above system, stirred at room temperature for 6 hours, TLC detected that the reaction was complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 300 mL ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent is dichloromethane : Methanol=20:1) Purify to obtain 5.1 g of reddish brown oil with a yield of 72%.

Example 16: (5R,10aR)-3-benzyloxycarbonyl-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrrolo[1 Preparation of ,2-a][1,5]diazaoctane-10a-carboxylic acid

Take 2g (3.6mmol) (5R, 10aR)-3-benzyloxycarbonyl-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrrolo [1,2-a][1,5] Diazaoctane-10a-methyl formate and 60mL methanol in a 250mL single-necked reaction flask, add 0.4g potassium hydroxide (7.2mmol), 60mL water, and heat to 50 After stirring for 3 hours at ℃, the reaction was complete as detected by TLC. 60mL of water was added, methanol was evaporated under reduced pressure, the remaining aqueous solution was extracted with ethyl acetate (2*100mL), the organic layer was discarded, the aqueous solution was adjusted to pH 1-2 with concentrated hydrochloric acid, and extracted with ethyl acetate (3*100mL), organic The layer was dried with sodium sulfate, filtered, and the solvent was distilled off under reduced pressure to obtain 1.9 g of oil, which was directly used in the next reaction.

Example 17: (5R,10aR)-3-benzyloxycarbonyl-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrrolo[1 Preparation of ,2-a][1,5]diazaoctane-10a-(N-diphenylmethyl)-carboxamide

Take (5R,10aR)-3-benzyloxycarbonyl-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrrolo[1,2- a] [1,5] Diazaoctane-10a-carboxylic acid 1.8g (3.3mmol), 2.6g PyBop (5.0mmol) in a 100mL single-mouth reaction flask, add 50mL DMF, take another 0.85g DIEA (6.6mmol) , Stir at room temperature for 30 minutes. Another 1.2 g (6.6 mmol) of benzhydrylamine was added to the system, heated to 90° C. and stirred for 12 hours. TLC detected that the reaction was complete. The reaction solution was poured into 300mL water, extracted with ethyl acetate (3*100mL), washed with saturated sodium chloride solution, the organic layer was dried with sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent is two Chloromethyl: methanol = 15:1) was purified to obtain 1.6 g of light yellow solid with a yield of 72%.

Example 18: (5R,10aR)-3-amino-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrrolo[1,2 -a] [1,5] Diazaoctane-10a-(N-diphenylmethyl)-formamide preparation

Take (5R,10aR)-3-benzyloxycarbonyl-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrrolo[1,2- a] [1,5] Diazaoctane-10a-(N-diphenylmethyl)-carboxamide 0.5g (0.70mmol) and 30mL methanol are dissolved in a 100mL single-necked reaction flask to obtain a colorless clear liquid, add 50mg (10%) Pd/C, stirred at room temperature for 12 hours under hydrogen atmosphere, the reaction was complete as detected by TLC. The Pd/C was removed by filtration, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane:methanol=15:1) to obtain 344 mg of pale yellow solid with a yield of 85%.

Example 19: (5R,10aR)-3-(isovaleryl)-amino-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one-dehydrogenation Preparation of pyrrolo[1,2-a][1,5]diazepine-10a-(N-diphenylmethyl)-carboxamide

Take 72 mg (0.7 mmol) of isovaleric acid and 10 mL of dichloromethane in a 100 mL single-necked reaction flask, add 342 mg HATU (0.9 mmol), 154 mg DIEA (1.2 mmol), and stir at room temperature for 30 minutes to obtain a light yellow clear liquid.

Take (5R,10aR)-3-amino-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrrolo[1,2-a] [1,5] Diazepine-10a-(N-diphenylmethyl)-carboxamide 344 mg (0.6 mmol) was dissolved in 10 mL of dichloromethane, added to the above system to obtain a yellow-green solution, stirred at room temperature 3 Hours, TLC detection, the reaction is complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 100 mL ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent was dichloromethane : Methanol=20:1) Purification to obtain 277 mg of white solid with a yield of 70%.

Example 20: (5R,10aR)-3-(isovaleryl)-amino-5-((R)-2-methylamino)-propionyl-amino-6-one-dehydropyrrolo[1,2 -a] [1,5] Diazaoctane-10a-(N-Diphenylmethyl)-Carboxamide) (I-12) Preparation

Take 277mg (0.42mmol) (5R,10aR)-3-(isovaleryl)-amino-5-((R)-2-(tert-butoxycarbonyl)methylamino)-propionyl-amino-6-one- Dehydropyrrolo[1,2-a][1,5]diazepine-10a-(N-diphenylmethyl)-carboxamide and 10mL absolute ethanol in a 100mL single-neck reaction flask, add 10mL hydrochloric acid Ethanol, stirring at room temperature for 2 hours, TLC detection, the reaction is complete. The solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain a colorless oily liquid. Add 100 mL ethyl acetate, adjust the pH to 7-8 with saturated sodium bicarbonate solution, separate the layers, dry the organic layer over sodium sulfate, filter, and distill off under reduced pressure Solvent, the residue was purified by column chromatography (eluent: dichloromethane: methanol = 15:1) to obtain 160 mg of white solid with a yield of 68%.

¹ H NMR (400MHz, d ₆ -DMSO): δ = 8.82-8.79 (m, 1H), 8.10-7.90 (m, 1H), 7.36-7.22 (m, 8H), 7.15-7.13 (m, 2H), 6.06-6.03 (m, 1H), 4.90-4.75 (m, 1H), 4.00-3.92 (m, 1H), 3.80-3.65 (m, 1H), 3.52 (s, 1H), 3.43-3.40 (m, 2H) ), 3.20-2.90 (m, 3H), 2.30-2.10 (m, 5H), 2.00-1.60 (m, 5H), 1.24 (s, 2H), 1.15-1.05 (m, 3H), 0.89-0.84 (m ,6H), [M+H] ⁺ :m/z=562.3

Example 21: (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-amino-6-one-dehydropyrrolo[1,2-a] [1,5] Preparation of Methyl Diazacyclo-10a-carboxylate

Take 6g (10.7mmol) (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-de Hydropyrrolo[1,2-a][1,5]diazepine-10a-methyl formate and 30mL of absolute ethanol are added to a 100mL single-neck reaction flask, add 30mL of hydrochloric acid ethanol, stir at room temperature for 2 hours, TLC detection , The reaction is complete. The solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 5.2 g of a colorless oily liquid, which was directly used in the next reaction.

Example 22: (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrolo[ Preparation of 1,2-a][1,5]diazepine-10a-methyl carboxylate

Take 1.2 g (11.8 mmol) of isovaleric acid and 20 mL of dichloromethane in a 100 mL single-necked reaction flask, add 6.1 g HATU (16.1 mmol), 2.8 g DIEA (21.4 mmol), and stir at room temperature for 30 minutes to obtain a yellow clear liquid.

Take (5R,10aR)3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-amino-6-one-dehydropyrrolo[1,2-a][1,5 ] 5.2 g of diazepine-10a-methyl formate was dissolved in 20 mL of dichloromethane, and added to the above system to obtain a yellow-green solution, which was stirred at room temperature for 3 hours, detected by TLC, and the reaction was complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 100 mL ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent was dichloromethane : Methanol=20:1) Purify to obtain 4.7 g of white solid with a yield of 81%.

Example 23: (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrolo[ Preparation of 1,2-a][1,5]diazaoctane-10a-carboxylic acid

Take 470mg (0.86mmol) (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrole And [1,2-a][1,5]diazepine-10a-methyl formate and 20mL methanol in a 100mL single-necked reaction flask, add 150mg potassium hydroxide (2.59mmol), 20mL water, and heat to 50 After stirring for 3 hours at ℃, the reaction was complete as detected by TLC. 20mL of water was added, methanol was evaporated under reduced pressure, the remaining aqueous solution was extracted with ethyl acetate (2*50mL), the organic layer was discarded, the aqueous solution was adjusted to pH 1-2 with concentrated hydrochloric acid, and extracted with ethyl acetate (3*60mL), organic The layer was dried over sodium sulfate, filtered, and the solvent was distilled off under reduced pressure to obtain 460 mg of oil, which was directly used in the next reaction.

Example 24: (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrolo[ Preparation of 1,2-a][1,5]diazepine-10a-(N-2-phenyl-benzyl)-carboxamide

Take (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrolo[1,2 -a] [1,5] Diazaoctane-10a-carboxylic acid 460mg, 670mg PyBop (1.29mmol) in a 100mL single-neck reaction flask, add 20mL DMF, another 221mg DIEA (1.72mmol), and stir at room temperature for 30 minutes. Another 173 mg (0.95 mmol) of 2-phenylbenzamide was added to the system, heated to 90° C. and stirred for 12 hours. TLC detected that the reaction was complete. The reaction solution was poured into 300 mL of water, extracted with ethyl acetate (3*100 mL), washed with saturated sodium chloride solution, the organic layer was dried with sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent is two Chloromethane: methanol = 15:1) was purified to obtain 358 mg of white solid with a yield of 60%.

Example 25: (5R,10aR)-3-((R)-2-methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrolo[1,2-a ] [1,5] Diazaoctane-10a-(N-2-phenyl-benzyl)-carboxamide) (I-02)

Take (5R,10aR)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrrolo[1,2 -a][1,5]diazepine-10a-(N-2-phenyl-benzyl)-carboxamide 358g (0.52mmol) and 20mL methanol in a 100mL single-necked reaction flask, dissolved into a colorless clear After adding 36mg (10%) Pd/C, stirring at room temperature for 12 hours under hydrogen atmosphere, TLC detected that the reaction was complete. The Pd/C was removed by filtration, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane:methanol=15:1) to obtain 256 mg of pale yellow solid, with a yield of 88%.

¹ H NMR (400MHz, d ₆ -DMSO): δ=8.43-8.42(m,1H), 7.97-7.95(m,1H),7.44-7.41(m,2H), 7.38-7.29(m,6H), 7.19-7.18(m,1H), 4.80-4.78(m,1H), 4.60-4.54(m,1H), 3.96-3.91(m,2H), 2.30-2.02(m,6H), 2.09-1.74(m ,6H),1.72-1.62(m,1H),1.05-1.03(d,2H),0.98-0.96(m,3H),0.83-0.78(m,6H),[M+H] ⁺ :m/z =562.3

Example 26

¹ H NMR (400MHz, d ₆ -DMSO): δ = 8.82-8.79 (m, 1H), 8.10-7.90 (m, 1H), 7.36-7.22 (m, 8H), 7.15-7.13 (m, 2H), 6.06-6.03 (m, 1H), 4.90-4.75 (m, 1H), 4.00-3.92 (m, 1H), 3.80-3.65 (m, 1H), 3.52 (s, 1H), 3.43-3.40 (m, 2H) ), 3.20-2.90 (m, 3H), 2.30-2.10 (m, 5H), 2.00-1.60 (m, 5H), 1.24 (s, 2H), 1.15-1.05 (m, 3H), 0.89-0.84 (m ,6H), [M+H] ⁺ :m/z=562.3

Example 27

H-NMR (400MHz, d ₆ -DMSO): δ=8.83-8.81 (d, 1H), 7.88-7.86 (d, 1H), 7.37-7.24 (m, 8H), 7.16-7.10 (m, 2H), 6.06-6.04 (d, 1H), 4.99-4.95 (m, 1H), 3.95-3.92 (m, 2H), 3.73-3.70 (m, 1H), 3.59-3.40 (m, 3H), 2.42-2.21 (m ,6H),2.05-1.92(m,3H),1.79-1.73(m,2H),1.34-1.18(m,6H),0.89-0.85(m,6H).[M+H] ⁺ :m/z =562.3

Example 28

¹ H NMR (400MHz, d ₆ -DMSO): δ = 8.57 (m, 1H), 8.03-7.95 (m, 1H), 7.31-7.27 (m, 2H), 7.22-7.21 (m, 3H), 4.93 4.79(m, 1H), 4.65-4.52(m, 1H), 4.05-3.93(m, 10H), 2.89(s, 2H), 2.73(s, 2H), 2.07(s, 2H), 2.02-1.98( s,12H),1.19-1.15(s,3H),1.00-0.98(m,2H),0.90-0.88(m,6H), [M+H] ⁺ :m/z=486.3

Example 29

¹ H NMR (400MHz, d ₆ -DMSO): δ=8.43-8.42(m,1H), 7.97-7.95(m,1H),7.44-7.41(m,2H), 7.38-7.29(m,6H), 7.19-7.18(m,1H), 4.80-4.78(m,1H), 4.60-4.54(m,1H), 3.96-3.91(m,2H), 2.30-2.02(m,6H), 2.09-1.74(m ,6H),1.72-1.62(m,1H),1.05-1.03(d,2H),0.98-0.96(m,3H),0.83-0.78(m,6H),[M+H] ⁺ :m/z =562.3

Example 30

H-NMR (400MHz, d ₆ -DMSO): δ = 8.77-8.75 (d, 1H), 7.94-7.92 (d, 1H), 7.37-7.34 (m, 2H), 7.19-7.15 (m, 4H), 7.11-7.06 (m, 2H), 6.08-6.06 (d, 1H), 4.94-4.92 (m, 1H), 3.99-3.92 (m, 2H), 3.72-3.52 (m, 1H), 3.48-3.41 (m ,3H), 2.50-2.41(m,1H), 2.18-2.13(m,6H),1.95-1.92(m,4H),1.25(s,2H),1.05-0.98(m,3H),0.87-0.84 (m,6H).[M+H] ⁺ :m/z=598.4

Example 31

H-NMR (400MHz, d ₆ -DMSO): δ = 8.80-8.78 (d, 1H), 7.90-7.88 (d, 1H), 7.34-7.32 (m, 4H), 7.27-7.23 (m, 4H), 7.11-7.09 (m, 2H), 6.03-6.01 (d, 1H), 4.92-5.05 (m, 1H), 3.98-3.92 (m, 1H), 3.86-3.68 (m, 1H), 3.52-3.40 (m ,1H),3.38-3.10(m,8H),2.42-2.21(m,2H),2.15-2.02(m,2H),1.79-1.73(m,2H),1.42-1.38(m,1H),1.34 -1.18(m,28H),1.12-0.95(m,1H),0.89-0.85(m,3H).[M+H] ⁺ :m/z=716.6

Example 32

H-NMR (400MHz, d ₆ -DMSO): δ = 8.82-8.80 (d, 1H), 7.98-7.95 (d, 1H), 7.33-7.30 (m, 4H), 7.29-7.21 (m, 9H), 7.18-7.13(m,1H), 6.08-6.06(d,1H), 4.99-4.96(m,1H), 3.98-3.95(m,1H), 3.79-3.75(m,1H), 3.26-3.24(m ,2H),3.21-3.12(m,2H),2.89-2.80(s,1H),2.79-2.70(m,3H),2.60-2.51(m,1H),2.32-2.26(m,2H),2.19 -2.15(s,3H),1.86-1.68(m,2H),1.36-1.30(m,1H),1.26-1.24(m,1H),1.07-0.98(m,4H).[M+H] ⁺ :m/z＝610.4

Example 33

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.83-8.81(d,1H), 8.17-8.15(m,1H), 7.58-6.85(m,13H), 6.07-6.05(d,1H) , 4.94-4.93(d, 1H), 4.16-4.14(m, 2H), 4.08-4.01(m, 2H), 3.95-3.92(d, 3H), 2.77-2.75(m, 2H), 2.39- 1.854( m,10H),1.07-1.02(m,3H).[M+H]+:m/z=600.2

Example 34

H-NMR (400MHz, d ₆ -DMSO): δ = 8.89-8.86 (d, 1H), 8.84-8.80 (d, 1H), 7.38-7.18 (m, 8H), 7.14-7.10 (m, 2H), 6.06-6.04(d,1H),5.00-4.95(m,1H),4.10-3.75(m,2H),3.08-3.00(s,3H),2.52-2.10(m,6H),2.09-2.07(m ,3H),1.11-1.38(m,3H),0.98-0.92(m,9H).[M+H] ⁺ :m/z=562.3

Example 35

H-NMR (400MHz, d ₆ -DMSO): δ=8.78-8.76(d,1H), 8.03-8.02(d,1H), 7.37-7.24(m,8H), 7.16-7.10(m,2H), 6.06-6.04 (d, 1H), 4.99-4.95 (m, 1H), 3.95-3.92 (m, 1H), 3.73-3.70 (m, 3H), 3.59-3.40 (m, 2H), 2.42-2.21 (m ,6H),2.05-1.92(m,2H),1.79-1.73(m,2H),1.34-1.18(m,3H),0.89-0.85(m,3H).[M+H] ⁺ :m/z =590.4

Example 36

¹ H-NMR (400MHz, d6-DMSO): δ = 8.31-8.29 (m, 2H), 8.02-8.00 (m, 1H), 7.92-7.88 (m, 1H), 7.29-7.18 (m, 10H), 4.79-4.93(m,4H),4.52-4.65(m,1H),3.93-4.05(m,10H),2.89(s,2H),2.73(s,2H),2.07(s,2H),1.98- 2.02(s,12H),1.15-1.19(s,3H),0.98-1.00(m,2H),0.88-0.90(m,6H),[M+H]+:m/z=500.4

Example 37

¹ HNMR (400MHz, d ₆ -DMSO): δ = 8.70-9.00 (m, 1H), 8.20-8.50 (m, 1H), 7.18-7.40 (m, 5H), 4.80-5.05 (m, 2H), 3.80 -4.20(m,2H),3.40-3.70(m,2H),2.98-3.18(m,1H),2.60-2.80(m,1H),2.30-2.40(m,1H),1.70-2.20(m, 10H),1.30-1.60(m,7H),1.10-1.30(s,6H),0.85(s,6H),[M+H] ⁺ :m/z=500.4

Example 38

¹ H NMR (400MHz, d ₆ -DMSO): δ = 9.58 (s, 1H), 8.95 (s, 1H), 8.47 (s, 1H), 8.10-8.02 (m, 1H), 7.78-7.68 (m, 2H),7.50(s,1H),7.44(s,1H),7.36-7.22(m,10H),7.15-7.13(m,2H),6.42-6.38(s,1H),6.10-6.01(s, 1H),4.75-4.90(s,1H),4.11-3.92(m,8H),3.82-3.75(s,1H),3.72-3.58(s,1H),3.15-2.92(m,1H),2.42- 2.38 (m, 1H), 2.10-2.00 (m, 6H), 1.96-1.89 (s, 2H), 1.78-1.62 (s, 1H), 1.60-1.53 (s, 1H), 1.51-1.48 (s, 1H) ),1.32-1.18(m,6H),1.15-1.05(m,3H),0.86-0.75(m,2H),[M+H] ⁺ :m/z=868.2

Example 39: Preparation of ethyl 4-p-toluenesulfonyloxybutyrate

Take 50g (0.38mol) of ethyl 4-hydroxybutyrate and 300mL of dichloromethane in a 1L single-necked reaction flask, add 76.8g (0.76mol) of triethylamine, cool to 5°C, and take 86.6g of p-toluenesulfonyl chloride ( 0.46 mol) was dissolved in 200 mL of dichloromethane and added dropwise to the above system. After the dropwise addition was completed, the mixture was stirred at room temperature for 6 hours, and TLC detected that the reaction was complete. Add 300mL of water to wash, wash with dilute hydrochloric acid (1M), wash with saturated brine, dry the organic layer with sodium sulfate, filter, distill off the solvent under reduced pressure, and column chromatography of the residue (eluent: ethyl acetate: petroleum ether = 1: 20) Purification to obtain 100 g of a colorless oily liquid with a yield of 92%.

Example 40: Preparation of ethyl 2-(tert-butoxycarbonyl)hydrazinobutyrate

Take 50g (0.17mol) of ethyl 4-p-toluenesulfonyloxybutyrate, 67g (0.51mol) of tert-butoxycarbonylhydrazine and 600mL of DMF in a 1L single-necked reaction flask, heat to 90℃ and stir for 6 hours, TLC detection, reaction complete. Add 2L water, extract with ethyl acetate (3*300mL), wash with saturated brine, dry the organic layer with sodium sulfate, filter, evaporate the solvent under reduced pressure, and column chromatography of the residue (eluent: ethyl acetate: petroleum ether = 1:10) Purification to obtain 25 g of colorless oily liquid with a yield of 60%.

Example 41: Preparation of ethyl 1-allyl-2-(tert-butoxycarbonyl)hydrazinobutyrate

Take 25g (0.10mol) of ethyl 2-(tert-butoxycarbonyl)hydrazinobutyrate and 300mL of DMF into a 500mL single-necked reaction flask, add 27.6g (0.20mol) of potassium carbonate, stir at room temperature, and take 14.5g of allyl bromide (0.12mol) was added to it, heated to 90°C and stirred for 12 hours, TLC detected, the reaction was complete. Add 1L water, extract with ethyl acetate (3*300mL), wash with saturated brine, dry the organic layer with sodium sulfate, filter, distill off the solvent under reduced pressure, and column chromatography of the residue (eluent: ethyl acetate: petroleum ether = 1:15) Purify to obtain 21.5 g of colorless oily liquid with a yield of 75%.

Example 42: Preparation of 1-allyl-2-(tert-butoxycarbonyl)hydrazino-2-bromo-butyric acid ethyl ester

Take 20g (70mmol) of ethyl 1-allyl-2-(tert-butoxycarbonyl)hydrazinobutyrate and 200mL of pre-dried tetrahydrofuran in a 500mL three-necked reaction flask, under nitrogen protection, cool to -60°C, take diisopropyl 140mL (1M in THF, 0.14mol) of lithium amide, add dropwise to it, keep stirring at -60°C for 2 hours, cool to -78°C, take 46g (0.14mol) of carbon tetrabromide dissolved in 100mL tetrahydrofuran, and add dropwise In the above system, the temperature was maintained at -78°C and stirred for 2 hours. TLC detected that the reaction was complete. The reaction was quenched by adding 20 mL of saturated ammonium chloride solution, washed with dilute hydrochloric acid (1M), washed with saturated brine, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent is ethyl acetate Ester: petroleum ether = 1:15) purified to obtain 20.7 g of a colorless oily liquid with a yield of 81%.

Example 43: Preparation of 1-allyl-hydrazino-2-bromo-butyric acid ethyl ester

Take 20g (54.8mmol) 1-allyl-2-(tert-butoxycarbonyl)hydrazino-2-bromo-butyric acid ethyl ester and 100mL absolute ethanol in a 500mL single-neck reaction flask, add 100mL hydrochloric acid ethanol, and stir at room temperature After 2 hours, TLC detection showed that the reaction was complete. The solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 16.2 g of a colorless oily liquid, which was directly used in the next reaction.

Example 44: Preparation of 1-allyl-pyrazole-3-carboxylic acid ethyl ester

Take 16.2g of 1-allyl-hydrazino-2-bromo-butyric acid ethyl ester and 200mL of DMF into a 500mL single-necked reaction flask, add 18.9g (0.14mol) of potassium carbonate, heat to 90℃ and stir for 12 hours, TLC detection, The reaction is complete. Add 600mL water, extract with ethyl acetate (3*300mL), wash with saturated brine, dry the organic layer with sodium sulfate, filter, distill off the solvent under reduced pressure, and column chromatography of the residue (eluent: ethyl acetate: petroleum ether = 1:10) Purification to obtain 6.3 g of white solid with a yield of 63%.

Example 45: Preparation of 1-allyl-2-((3-benzyloxycarbonyl-amino-2-tert-butoxycarbonyl-amino)propionyl)-pyrazole-3-carboxylic acid ethyl ester

Take 13.9g (41mmol) of (S)-2-(Boc-amino)-3-(Cbz-amino)propionic acid and 200mL of dichloromethane in a 500mL single-mouth reaction flask, add 20.8g HATU (54.7mmol), 8.8g DIEA (68.4mmol), stirred at room temperature for 1 hour to obtain a yellow clear liquid.

In addition, 6.3 g (34.2 mmol) of ethyl 1-allyl-pyrazole-3-carboxylate was dissolved in 100 mL of dichloromethane, and added dropwise to the above-mentioned yellow clear liquid, and stirred at room temperature for 6 hours to obtain a reddish-brown solution. The reaction was detected by TLC complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 400 mL ethyl acetate, washed with 1M HCl solution, saturated sodium bicarbonate solution, saturated sodium chloride solution, the organic layer was dried with sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The product was purified by column chromatography (eluent: ethyl acetate: petroleum ether = 1:2) to obtain 14.6 g of white solid with a yield of 85%.

Example 46: Preparation of 1-acetaldehyde-2-((3-benzyloxycarbonyl-amino-2-tert-butoxycarbonyl-amino)propionyl)-pyrazole-3-carboxylic acid ethyl ester

Take 1-allyl-2-((3-benzyloxycarbonyl-amino-2-tert-butoxycarbonyl-amino)propionyl)-pyrazole-3-carboxylic acid ethyl ester 14.6g (29mmol) and 200mL dichloromethane In a 500mL three-necked reaction flask, cool to -78°C, and pass ozone in for about 30 minutes until the system turns blue. Stir at -78°C for 30 minutes and stop passing ozone. Nitrogen was bubbled through to remove excess ozone until the blue color disappeared, and a colorless solution was obtained. 100 mL of triethylamine was added, returned to room temperature, and stirred for 2 hours. TLC detected that the reaction was complete. Wash with 1M citric acid, wash with saturated sodium chloride aqueous solution, dry the organic layer with sodium sulfate, filter, distill off the solvent under reduced pressure, and purify the residue by column chromatography (eluent: ethyl acetate: petroleum ether = 1:2) , To obtain 11.5 g of white solid with a yield of 78%.

Example 47: (5S,8S)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[1,2-a][1,2,5]triazaoctane Preparation of -8-methyl formate

Take 11.5g (22.7mmol) 1-acetaldehyde-2-((3-benzyloxycarbonyl-amino-2-tert-butoxycarbonyl-amino)propionyl)-pyrazole-3-carboxylic acid ethyl ester and 200mL methanol in Dissolve in a 500 mL single-mouth reaction flask to obtain a colorless clear solution, add 1.2 g (10%) Pd/C, and stir at room temperature for 12 hours under a hydrogen atmosphere. TLC detects that the reaction is complete. The Pd/C was removed by filtration, the solvent was distilled off under reduced pressure, 200 mL of tetrahydrofuran was added to dissolve, and 2.1 g (34.1 mmol) of sodium cyanoborohydride was added, stirred at room temperature, and TLC detected that the reaction was complete. The reaction was quenched by adding 20 mL of water, filtered, and the solvent was evaporated under reduced pressure. The residue was dissolved in 200 mL of ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. After purification (eluent: dichloromethane: methanol = 10:1), 6.2 g of white solid was obtained with a yield of 80%.

Example 48: (5S,8S)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyridine Preparation of azolo[1,2-a][1,2,5]triazaoctane-8-methyl carboxylate

Take 2.6g (10.9mmol) of N-Cbz-N-methyl-D-alanine and 80mL of dichloromethane in a 250mL single-mouth reaction flask, add 5.2g HATU (13.6mmol), 2.4gDIEA (18.1mmol), room temperature Stir for 1 hour to obtain a yellow clear liquid.

Take (5S,8S)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[1,2-a][1,2,5]triazaoctane-8- 3.1 g (9.1 mmol) of methyl formate was dissolved in 50 mL of dichloromethane, added dropwise to the above system, stirred at room temperature for 6 hours, TLC detected that the reaction was complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 300 mL ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent is dichloromethane : Methanol=20:1) Purification to obtain 3.7 g of brown oil with a yield of 72%.

Example 49: (5S,8S)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyridine Preparation of azolo[1,2-a][1,2,5]triazaoctane-8-carboxylic acid

Take 1g (1.8mmol) (5S, 8S)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-de Hydropyrazolo[1,2-a][1,2,5]triazaoctane-8-methyl carboxylate and 30mL methanol in a 250mL single-neck reaction flask, add 0.3g (5.4mmol) of potassium hydroxide, 30mL of water, heated to 50°C, stirred for 3 hours, TLC detected that the reaction was complete. 60mL of water was added, methanol was evaporated under reduced pressure, the remaining aqueous solution was extracted with ethyl acetate (2*100mL), the organic layer was discarded, the aqueous solution was adjusted to pH 1-2 with concentrated hydrochloric acid, and extracted with ethyl acetate (3*100mL), organic The layer was dried with sodium sulfate, filtered, and the solvent was distilled off under reduced pressure to obtain 0.98 g of oil, which was directly used in the next reaction.

Example 50: (5S,8S)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyridine Preparation of azolo[1,2-a][1,2,5]triazaoctane-8-(N-diphenylmethyl)-carboxamide

Take (5S,8S)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[ 1,2-a][1,2,5]Triazaoctane-8-carboxylic acid 0.98g (1.8mmol), 1.4g PyBop (2.7mmol) in a 100mL single-mouth reaction flask, add 50mL DMF, and another 0.46 g DIEA (3.6mmol), stir at room temperature for 30 minutes. Another 0.66 g (3.6 mmol) of benzhydrylamine was added to the system, heated to 90° C., and stirred for 12 hours. TLC detected that the reaction was complete. The reaction solution was poured into 300mL water, extracted with ethyl acetate (3*100mL), washed with saturated sodium chloride solution, the organic layer was dried with sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent is two Chloromethane: methanol = 15:1) was purified to obtain 0.77 g of light yellow solid with a yield of 60%.

Example 51: (5S,8S)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-amino-6-one-dehydropyrazolo[1,2-a ][1,2,5]Triazaoctane-8-(N-diphenylmethyl)-carboxamide

Take 0.5g (0.7mmol) (5S, 8S) 3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(tert-butyloxycarbonyl)-amino-6-one-de Hydropyrazolo[1,2-a][1,2,5]triazaoctane-8-(N-diphenylmethyl)-formamide and 10mL of absolute ethanol are added to a 100mL single-mouth reaction flask 20 mL of hydrochloric acid ethanol, stirred at room temperature for 2 hours, TLC detection, the reaction was complete. The solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 0.45 g of a colorless oily liquid, which was directly used in the next reaction.

Example 52: (5S,8S)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrazolo Preparation of [1,2-a][1,2,5]Triazaoctane-8-(N-Diphenylmethyl)-carboxamide

Take 82 mg (0.8 mmol) of isovaleric acid and 10 mL of dichloromethane in a 100 mL single-necked reaction flask, add 0.42 g HATU (1.1 mmol), 0.18 g DIEA (1.4 mmol), and stir at room temperature for 30 minutes to obtain a light yellow clear liquid.

Take (5S,8S)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-amino-6-one-dehydropyrazolo[1,2-a][1 ,2,5]Triazaoctane-8-(N-diphenylmethyl)-carboxamide 0.45g was dissolved in 10mL of dichloromethane, added to the above system to obtain a yellow-green solution, stirred at room temperature for 3 hours, TLC The reaction is complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 100 mL ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent was dichloromethane : Methanol=20:1) Purification to obtain 345 mg of white solid with a yield of 71%.

Example 53: (5S,8S)-3-((R)-2-methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrazolo[1,2- a][1,2,5]Triazaoctane-8-(N-diphenylmethyl)-carboxamide (I-13)

Take (5S,8S)-3-((R)-2-(benzyloxycarbonyl)methylamino)-propionyl-5-(isovaleryl)-amino-6-one-dehydropyrazolo[1, 2-a][1,2,5]Triazaoctane-8-(N-diphenylmethyl)-carboxamide 200mg (0.29mmol) and 50mL methanol in a 100mL single-necked reaction flask, dissolved into colorless clear After adding 20mg (10%) Pd/C, stirring at room temperature for 12 hours under a hydrogen atmosphere, TLC detected that the reaction was complete. Pd/C was removed by filtration, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane:methanol=15:1) to obtain 138 mg of white solid with a yield of 85%.

¹ H NMR (400MHz, d ₆ -DMSO): δ=8.03-8.00 (m, 1H), 7.49-7.32 (m, 2H), 7.30-7.22 (m, 4H), 7.19-7.10 (m, 4H), 6.06-6.03 (m, 1H), 5.05-4.99 (m, 1H), 4.45-4.39 (m, 1H), 3.80-3.62 (m, 1H), 3.52 (s, 1H), 3.43-3.40 (m, 2H) ), 3.20-2.90 (m, 3H), 2.78-2.47 (m, 2H), 2.30-2.10 (m, 5H), 2.00-1.60 (m, 5H), 1.18-1.05 (m, 3H), 0.89-0.82 (m,6H), [M+H] ⁺ :m/z=563.3

Example 54: (5S,8S)-3-(isovaleryl)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[1,2-a][1,2 ,5] Preparation of methyl triazaoctane-8-carboxylate

Take 1.1 g (11 mmol) of isovaleric acid and 30 mL of dichloromethane in a 100 mL single-neck reaction flask, add 5.7 g HATU (15 mmol), 2.6 g DIEA (20 mmol), and stir at room temperature for 30 minutes to obtain a light yellow clear liquid.

Take (5S,8S)5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[1,2-a][1,2,5]triazaoctane-8-carboxylic acid 3.4 g (10 mmol) of methyl ester was dissolved in 30 mL of dichloromethane, and added to the above system to obtain a yellow-green solution. The mixture was stirred at room temperature for 3 hours, detected by TLC, and the reaction was complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 100 mL ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent was dichloromethane : Methanol=20:1) Purify to obtain 2.8 g of white solid with a yield of 66%.

Example 55: (5S,8S)-3-(isovaleryl)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[1,2-a][1,2 ,5] Preparation of triazaoctane-8-carboxylic acid

Take 1g (2.3mmol) (5S,8S)-3-(isovaleryl)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[1,2-a][1 ,2,5]Triazaoctane-8-methyl formate and 30mL methanol in a 100mL single-necked reaction flask, add 0.3g (5.9mmol) of potassium hydroxide, 30mL of water, heat to 50℃, stir for 3 hours, TLC The detection reaction is complete. 30mL of water was added, methanol was evaporated under reduced pressure, the remaining aqueous solution was extracted with ethyl acetate (2*100mL), the organic layer was discarded, the aqueous solution was adjusted to pH 1-2 with concentrated hydrochloric acid, and extracted with ethyl acetate (3*100mL). The layer was dried with sodium sulfate, filtered, and the solvent was distilled off under reduced pressure to obtain 0.95 g of oil, which was directly used in the next reaction.

Example 56: (5S,8S)-3-(isovaleryl)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[1,2-a][1,2 ,5] Preparation of triazaoctane-8-(N-diphenylmethyl)-carboxamide

Take (5S,8S)-3-(isovaleryl)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[1,2-a][1,2,5] Triazaoctane-8-carboxylic acid 0.95g (2.3mmol), 1.8g PyBop (3.6mmol) in a 100mL single-necked reaction flask, add 50mL DMF, another 0.60g DIEA (4.6mmol), and stir at room temperature for 30 minutes. Another 0.84 g (4.6 mmol) of benzhydrylamine was added to the system, heated to 90° C. and stirred for 12 hours. TLC detected that the reaction was complete. The reaction solution was poured into 300mL water, extracted with ethyl acetate (3*100mL), washed with saturated sodium chloride solution, the organic layer was dried with sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent is two Chloromethane: methanol = 15:1) was purified to obtain 0.70 g of light yellow solid with a yield of 53%.

Example 57: (5S,8S)-3-(isovaleryl)-5-amino-6-one-dehydropyrazolo[1,2-a][1,2,5]triazaoctane Preparation of -8-(N-diphenylmethyl)-formamide

Take 200mg (0.35mmol) (5S,8S)-3-(isovaleryl)-5-(tert-butyloxycarbonyl)-amino-6-one-dehydropyrazolo[1,2-a][1 ,2,,5]Triazaoctane-8-(N-diphenylmethyl)-carboxamide and 10mL of absolute ethanol were added to a 100mL single-necked reaction flask, 20mL of hydrochloric acid ethanol was added, stirred at room temperature for 2 hours, TLC detection, The reaction is complete. The solvent and excess hydrogen chloride were distilled off under reduced pressure to obtain 0.17 g of a colorless oily liquid, which was directly used in the next reaction.

Example 58: (5S,8S)-3-(isovaleryl)-5-((S)-2-(benzyloxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrazolo Preparation of [1,2-a][1,2,5]Triazaoctane-8-(N-Diphenylmethyl)-carboxamide

Take 99mg (0.42mmol) of N-Cbz-N-methyl-D-alanine and 20mL of dichloromethane in a 100mL single-mouth reaction flask, add 0.24g HATU (0.63mmol), 0.12g DIEA (0.88mmol), stir at room temperature In 1 hour, a yellow clear liquid was obtained.

Take (5S,8S)-3-(isovaleryl)-5-amino-6-one-dehydropyrazolo[1,2-a][1,2,5]triazaoctane-8- 0.17 g of (N-diphenylmethyl)-formamide was dissolved in 20 mL of dichloromethane, added dropwise to the above system, and stirred at room temperature for 6 hours. TLC detected that the reaction was complete. The solvent was evaporated under reduced pressure, the residue was dissolved in 100 mL ethyl acetate, washed with saturated sodium chloride solution, the organic layer was dried over sodium sulfate, filtered, the solvent was evaporated under reduced pressure, and the residue was column chromatographed (eluent was dichloromethane : Methanol=20:1) Purification to obtain 160 mg of brown oil with a yield of 66%.

Example 59: (5S,8S)-3-(isovaleryl)-5-((S)-2-methylamino)-propionyl-amino-6-one-dehydropyrazolo[1,2- a][1,2,5]Triazaoctane-8-(N-diphenylmethyl)-carboxamide (I-14)

Take (5S,8S)-3-(isovaleryl)-5-((S)-2-(benzyloxycarbonyl)methylamino)-propionyl-amino-6-one-dehydropyrazolo[1, 2-a][1,2,5]Triazaoctane-8-(N-diphenylmethyl)-carboxamide 160mg (0.23mmol) and 50mL methanol in a 100mL single-necked reaction flask, dissolve to a colorless clear After adding 16mg (10%) Pd/C, stirring at room temperature for 12 hours under hydrogen atmosphere, TLC detected that the reaction was complete. Pd/C was removed by filtration, the solvent was distilled off under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane:methanol=15:1) to obtain 106 mg of white solid with a yield of 82%.

¹ H NMR (400MHz, d ₆ -DMSO): δ=8.05-8.02 (m, 1H), 7.47-7.29 (m, 2H), 7.31-7.12 (m, 4H), 7.18-7.08 (m, 4H), 6.06-6.03(m,1H),5.00-4.93(m,1H),4.42-4.39(m,1H),3.92-3.73(m,1H),3.54(s,1H),3.41-3.39(m,2H) ), 3.22-2.88 (m, 3H), 2.70-2.33 (m, 2H), 2.21-2.12 (m, 5H), 2.05-1.78 (m, 5H), 1.28-1.15 (m, 3H), 0.92-0.85 (m,6H), [M+H] ⁺ :m/z=563.3

Example 60:

¹ H NMR (400MHz, d ₆ -DMSO): δ=8.03-8.00 (m, 1H), 7.49-7.32 (m, 2H), 7.30-7.22 (m, 4H), 7.19-7.10 (m, 4H), 6.06-6.03 (m, 1H), 5.05-4.99 (m, 1H), 4.45-4.39 (m, 1H), 3.80-3.62 (m, 1H), 3.52 (s, 1H), 3.43-3.40 (m, 2H) ), 3.20-2.90 (m, 3H), 2.78-2.47 (m, 2H), 2.30-2.10 (m, 5H), 2.00-1.60 (m, 5H), 1.18-1.05 (m, 3H), 0.89-0.82 (m,6H), [M+H] ⁺ :m/z=563.3

Example 61:

¹ H NMR (400MHz, d ₆ -DMSO): δ=8.05-8.02 (m, 1H), 7.47-7.29 (m, 2H), 7.31-7.12 (m, 4H), 7.18-7.08 (m, 4H), 6.06-6.03(m,1H),5.00-4.93(m,1H),4.42-4.39(m,1H),3.92-3.73(m,1H),3.54(s,1H),3.41-3.39(m,2H) ), 3.22-2.88 (m, 3H), 2.70-2.33 (m, 2H), 2.21-2.12 (m, 5H), 2.05-1.78 (m, 5H), 1.28-1.15 (m, 3H), 0.92-0.85 (m,6H), [M+H] ⁺ :m/z=563.3

Example 62:

¹ HNMR (400MHz, d ₆ -DMSO): δ=8.04-8.01 (m, 1H), 7.52-7.36 (m, 1H), 7.27-7.21 (m, 2H), 7.16-7.05 (m, 2H), 6.05 -6.01(m,1H),5.04-4.97(m,1H),4.52-4.43(m,1H),4.00-3.92(m,3H),3.84-3.67(m,1H),3.58(s,1H) ,3.45-3.41(m,2H),3.27-2.93(m,3H),2.74-2.53(m,2H),2.35-2.18(m,5H),2.12-1.68(m,5H),1.21-1.09( m,3H),0.93-0.88(m,6H),[M+H] ⁺ :m/z=563.3

Example 63:

¹ H-NMR (400MHz, d ₆ -DMSO): δ=8.12-8.05 (m, 1H), 7.41-7.39 (m, 2H), 7.32-7.21 (m, 6H), 7.13-7.06 (m, 1H) ,6.11-6.07(m,1H),5.07-4.97(m,1H),4.46-4.37(m,1H),3.81-3.64(m,1H),3.55(s,1H),3.46-3.41(m, 2H), 3.25-2.96 (m, 3H), 2.73-2.49 (m, 2H), 2.33-2.17 (m, 5H), 2.06-1.68 (m, 5H), 1.12-1.09 (m, 3H), 0.92- 0.88(m,6H), [M+H] ⁺ :m/z=563.6

Example 64:

¹ H NMR (400MHz, d ₆ -DMSO): δ=8.08-8.02 (m, 1H), 7.44-7.38 (m, 1H), 7.32-7.26 (m, 2H), 7.19-7.15 (m, 4H), 7.11-7.06(m,2H),6.06-6.03(m,1H),5.06-4.97(m,1H),4.44-4.35(m,1H),3.82-3.61(m,1H),3.58(s,1H) ), 3.47-3.42 (m, 2H), 3.23-2.96 (m, 3H), 2.73-2.41 (m, 2H), 2.38-2.12 (m, 5H), 2.07-1.66 (m, 5H), 1.11-1.08 (m,3H),0.95-0.86(m,6H),[M+H] ⁺ :m/z=599.4

Example 65: Test of the binding affinity of the compound to XIAP, cIAP1, and cIAP2

In order to test the binding affinity of the compounds of the present invention to XIAP, cIAP1, and cIAP2 proteins, a fluorescence polarization (FP)-based competitive assay method was used for the assay. For the cIAP1-BIR3 and cIAP2-BIR3 assay methods, fluorescently labeled Smac mimics ( Smac-2F) is used as a fluorescent probe. The K _d values of Smac-2F, cIAP1-BIR3, and cIAP2-BIR3 are determined by monitoring the total fluorescence polarization of a mixture composed of a solid concentration fluorescent probe and an increasing concentration of protein up to complete saturation. The fluorescence polarization value was measured in a Microfluor 2 96-well black round bottom plate (Fisher Scientific) using an Infinite M-1000 microplate reader (Tecan US, Research Triangle Park, NC). In 96-wells, add 1nM Smac-2F and increasing concentrations of protein to the assay buffer (100mM potassium phosphate, pH=7.5; 100ug/ml bovine gamma globulin; 0.02% sodium azide, Invitrogen), and finally The volume is 125ul. The plates were mixed on a shaker for 15 minutes and incubated at room temperature for 1-3 hours. The polarization value was measured in millipolarization unit at 485nm excitation wavelength and 530nm emission wavelength. Then use GraphPad Prism 5.0 software to calculate the equilibrium dissociation constant (K _d ) by fitting the S-shaped dose-dependent FP increase as a function of protein concentration

The K _i value of the compound is determined by a dose-dependent competitive binding experiment, in which a serial dilution of the compound competes with a fixed concentration of fluorescent probe to bind to a fixed concentration of protein (usually the Kd value measured above is 2-3 times) . 5ul test compound in DMSO and 120ul pre-incubated protein/tracer in assay buffer solution (100mM potassium phosphate, pH=7.5; 100ug/ml bovine gamma globulin; 0.02% sodium azide, Invitrogen) The complex mixture is added to the assay plate and incubated at room temperature for 1-3 hours. The final concentrations of protein and probe were 3nm and 1nm, 5nm and 1nm for cIAP1-BIR3 and cIAP2-BIR3, respectively. A negative control containing only protein/probe complexes (equivalent to 0% inhibition) and a positive control containing only free probes (equivalent to 100% inhibition) were included in each assay plate. Measure the EP value as described above. The IC50 value was determined by nonlinear regression fitting of the competition curve. The Ki value of the competitive inhibitor is calculated using the previously described derivation equation based on the measured IC50 value, the Kd value of the probe and different proteins, and the concentration of the protein and probe in the refining assay.

The FP-based assay for the XIAP-BIR2-BIR3 protein was performed using the same procedure. In this measurement, a bivalent fluorescent-tagged peptide Smac mimic (Smac-1F) was used as a fluorescent probe, and the Kd value of XIAP-BIR2-BIR3 was measured similarly to the saturation experiment. 0.01% Triton X-100 was added to the assay buffer to achieve the stable fluorescence and polarization values of the dimer fluorescent probe. The final protein and probe concentrations used in the competition assay were 2nm and 1nm, respectively. The test results of the compounds of the present invention are shown in the following table.

The experimental results found that the compound of the present invention has good binding affinity with XIAP, cIAP1, and cIAP2 proteins, and has good IAP inhibitory activity.

Example 67: Cell growth inhibition test in MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines

The effects of the compounds of the invention on the growth of different cancer cell lines were tested. The cells were seeded on a 96-well flat-bottom cell culture plate together with the test compound at a density of 3000 cells/well and the cells were cultured at 37°C in an atmosphere of 95% air and 5% CO ₂ for 4 days. Use WST-8 kit and 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H- Tetrazolium monosodium salt was used to determine the cell growth inhibition rate after treatment with different concentrations of the compound. WST-8 was added to each well at a final concentration of 10%, and then the plate was incubated at 37°C for 2-3 hours. The absorbance of the sample was measured at 450 nm using a ULTRATecan reader (Molecular Device). By comparing the absorbance of untreated cells and the treated cells with the test compound, the test compound is calculated inhibitory concentration (IC ₅₀₎ 50% of cell growth. The test results are shown in the table below.

The compound of the present invention has a good inhibitory effect on cell growth in MDA-MB-231 breast cancer and PC-3 pancreatic cancer cell lines.

Claims (12)

1. A compound or a pharmaceutically acceptable salt or isomer thereof, characterized in that the compound has the following general formula:

   

   Wherein, X represents a C or N atom, when X is a C atom, Y is a CONHR ₃ and Z represents a H atom; when X is a N atom, Z is a CONHR ₃ and Y does not exist;

   The R ₁ and R ₂ are each independently selected from C _1-20 alkylamino groups, C _5-8 cycloalkylamino groups,

   

   C _1～20 alkyl group,

   

   

   Or α-amino acid residues,

   The α-amino acid residue is a substituent formed by the deletion of the carboxyl group on the α-carbon of the α-amino acid, and the Ar group represents an aryl or heteroaryl group with 5-8 atoms. The m and n are each independently selected from any integer of 1 to 3;

   R _{3 is} selected from substituted or unsubstituted benzyl, and the substituent is optionally selected from phenyl, halogen, C _1-4 alkyl or p-fluorophenyl.
2. A compound or a pharmaceutically acceptable salt or isomer thereof, characterized in that the compound has the following general formula:

   

   Wherein, in the above formula I or II,

   Said R _{1 is} selected from C _1-20 alkylamino groups,

   

   C _1～20 alkyl group,

   

   

   Or any of the α-amino acid residues,

   R _{2 is} selected from C _1-20 alkylamino group, C _5-8 cycloalkylamino group, C _1-20 alkyl group,

   

   

   Or any of the α-amino acid residues,

   The α-amino acid residue is a substituted residue formed by the deletion of the carboxyl group on the α-carbon in the α-amino acid, and the Ar group represents an aryl group and a heteroaryl group with 5-8 atoms. , The m and n are each independently selected from any integer of 1 to 3,

   R _{3 is} selected from substituted or unsubstituted benzyl, and the substituent is optionally selected from phenyl, halogen, C _1-4 alkyl or p-fluorophenyl.
3. The compound and salt or isomer thereof according to claim 2, wherein the alkylamino group in formula I or II is

   

   And its isomers.
4. The compound and salt or isomer thereof according to claim 2, wherein the C _5-6 cycloalkylamino group in formula I or II is

   
5. The compound and its salt or isomer according to claim 2, wherein the α-amino acid residue is

   

   or

   
6. The compound according to claim 2 and its salt or isomer, characterized in that R ₂ in formula I or II is selected from C _1-13 alkyl.
7. The compound and its salt or isomer according to claim 2, characterized in that the aryl group in formula I or II is phenyl.
8. The compound and its salt or isomer according to claim 2, characterized in that the heteroaryl group in formula I or II is imidazolyl.
9. A compound characterized in that its structure is selected from

   

   

   

   

   
10. A pharmaceutical composition characterized by containing a therapeutically effective amount of the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt or isomer thereof as an active ingredient, and one or more pharmacological agents Acceptable carriers, diluents and/or excipients.
11. The use of the compound described in any one of claims 1 to 9 or a pharmaceutically acceptable salt or isomer thereof in the preparation of a medicine for diseases related to IAP protein.
12. Use of a compound containing the compound described in any one of claims 1 to 9 or a pharmaceutically acceptable salt or isomer thereof in the preparation of a medicament for treating all types of cancers that have not undergone apoptosis.