An MDM2 inhibitor, and preparation method, pharmaceutical composition and application thereof

Abstract

The present application provides a MDM2 inhibitor, a preparation method, a pharmaceutical composition and an application thereof. Specifically, the present application provides a compound as shown in formula I, wherein the definitions of each group are described in the description. The compound has excellent MDM2 inhibitory activity, and thus can be used for preparing a pharmaceutical composition for treating cancer (particularly, p53 wild-type cancer) and other diseases related to MDM2 activity.

Description

Technical Field

[0001] The present invention relates to the field of small molecule drugs. Specifically, the present invention provides an MDM2 inhibitor, and a preparation method, a pharmaceutical composition and application thereof. Background Art

[0002] The p53 gene is the tumor suppressor gene most closely associated with human tumors. It maintains genomic stability and inhibits or prevents cellular transformation, thereby suppressing tumorigenesis. The search for new anti-tumor drugs targeting p53 has become a hot topic in this field. Research has shown that MDM2 (murine double minute 2) is a key negative regulator of p53. p53 activates MDM2 transcription, which in turn inhibits p53 activity, forming an autoregulatory feedback loop that maintains low levels of both p53 and MDM2 under normal circumstances. Aberrant expression of MDM2 in tumor cells leads to rapid degradation of p53 and inactivation of the p53 pathway, thereby compromising its tumor suppressive properties. Consequently, releasing p53 from MDM2 control and activating the p53 pathway are expected to inhibit tumor cell growth and induce apoptosis. Unlike p53 activation in normal cells due to less common causes, tumor cells are under constant cellular stress, including hypoxia and activation of pro-apoptotic oncogenes. Thus, inactivation of the p53 pathway in tumors has a strong selective advantage, and some researchers have proposed that eliminating p53 function may be a prerequisite for tumor survival. In support of this notion, three research groups have used mouse models to demonstrate that loss of p53 function is a persistent requirement for tumor maintenance. When the researchers restored p53 function in tumors with p53 inactivation, the tumors regressed.

[0003] In 50% of solid tumors and 10% of liquid tumors, p53 is inactivated through mutation and / or deletion. Other key members of the p53 pathway also undergo genetic or epigenetic alterations in cancer. MDM2, an oncoprotein, inhibits p53 function and has been reported to be activated by gene amplification at a rate of up to 10%. MDM2 is in turn inhibited by another tumor suppressor, p14ARF. Alterations downstream of p53 are thought to be responsible for at least partial inactivation of the p53 pathway in p53 wild-type tumors. In support of this concept, some p53 wild-type tumors appear to exhibit reduced apoptotic function, yet their ability to undergo cell cycle arrest remains intact. One cancer treatment strategy involves the use of small molecules that bind to MDM2 and counteract its interaction with p53. MDM2 inhibits p53 activity through three mechanisms: 1) acting as an E3 ubiquitin ligase to promote p53 degradation; 2) binding to and blocking the p53 transcriptional activation domain; and 3) exporting p53 from the nucleus to the cytoplasm. All three mechanisms are blocked by counteracting the MDM2-p53 interaction. This therapeutic strategy could be targeted at p53 wild-type tumors, and studies have shown promise in reducing tumor growth in vitro and in vivo using small-molecule MDM2 inhibitors. Furthermore, in patients with p53-inactivated tumors, MDM2 inhibition could selectively protect normal tissues from damage, as wild-type p53 is stabilized in normal tissues following MDM2 inhibition.

[0004] In summary, there is a need in the art to develop novel MDM2 inhibitors. Summary of the Invention

[0005] The purpose of the present invention is to provide a novel MDM2 inhibitor.

[0006] In the first aspect of the present invention, the present invention provides a compound represented by the following formula I, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, or a solvate thereof:

[0007]

[0008] in,

[0009] is a 5-membered, 6-membered or 7-membered heterocyclic group; wherein the heterocyclic group includes 1-3 N atoms and 0-2 heteroatoms selected from the group consisting of S and O;

[0010] X is C=O or S=(O)2;

[0011] n is 1, 2, 3, or 4;

[0012] Each R is independently selected from the following group: H, cyano, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C 10 Aryl, or a substituted or unsubstituted 5-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O;

[0013] Z1 is selected from the following groups: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl (including monocyclic, cyclic or bridged forms), substituted or unsubstituted C6-C 10 aryl;

[0014] Q is selected from the following group:

[0015] m and p are each independently 1, 2, 3 or 4;

[0016] Each Z2 or Z3 is independently selected from the group consisting of none, substituted or unsubstituted C1-C7 alkylene, NR1, O, S, C=O, S=(O)2;

[0017] Z4 is selected from Wherein, said R1 is selected from the following groups: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C 10 Aryl, cyano, -C(=O)-NRdRe, -C(=O)-substituted or unsubstituted C1-C6 alkoxy, -C(=O)-substituted or unsubstituted C1-C6 alkyl, -C(=O)-substituted or unsubstituted C3-C10 cycloalkyl, -C(=O)-substituted or unsubstituted C2-C6 alkenyl, -C(=O)-substituted or unsubstituted C2-C6 alkynyl;

[0018] Rd and Re are each independently selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C6-C 10 Aryl; or said Rd and Re and the adjacent N atom form a 4-10 membered heterocyclic ring containing 1-2 nitrogen atoms and 0-2 S or O atoms;

[0019] R2 is selected from the following groups: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C 10 Aryl, or a substituted or unsubstituted 5-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O;

[0020] Unless otherwise specified, the term "substituted" refers to substituted by one or more (e.g., 2, 3, 4, etc.) substituents selected from the group consisting of halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo, -CN, hydroxy, amino, carboxyl, a group selected from the group consisting of C6-C10 aryl, halogenated C6-C10 aryl, a 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, and a halogenated 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O; the substituents are selected from the group consisting of halogen, C1-C6 alkoxy;

[0021] The additional condition is that when Z4 is When Q is

[0022] In another preferred embodiment, the Select from the following groups:

[0023] In another preferred embodiment, the compound of formula I has the structure described in formula II below:

[0024]

[0025] In another preferred embodiment, n is 3.

[0026] In another preferred embodiment, R is selected from the following groups: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C 10 Aryl.

[0027] In another preferred embodiment, the compound of formula I has the structure described in formula III below:

[0028]

[0029] wherein Ra and Rb are each independently substituted or unsubstituted C6-C 10 Aryl, or a substituted or unsubstituted 5-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O;

[0030] Rc is selected from the following groups: H, cyano, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy;

[0031] The definitions of the groups are as described above.

[0032] In another preferred embodiment, Ra and Rb are each independently a substituted or unsubstituted phenyl group. In another preferred embodiment, the compound of formula I has the structure described in formula IV below:

[0033]

[0034] In another preferred embodiment, the compound of formula I has a structure selected from the following Table A:

[0035] Table A

[0036]

[0037]

[0038]

[0039]

[0040]

[0041]

[0042]

[0043] In another preferred embodiment, the "isomer" refers to the chiral center optically active isomers.

[0044] In the second aspect of the present invention, there is provided a method for preparing the compound of formula I described in the first aspect of the present invention, the method comprising or being carried out by the following steps (1), (2) or (3):

[0045] Step (1):

[0046]

[0047] Step (2):

[0048]

[0049] Step (3):

[0050]

[0051] Wherein, the definitions of each group are as described in the first aspect.

[0052] In the third aspect of the present invention, a pharmaceutical composition is provided, which comprises (1) the compound described in the first aspect of the present invention or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate; and (2) a pharmaceutically acceptable carrier.

[0053] In the fourth aspect of the present invention, provided is the use of the compound as described in the first aspect of the present invention or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition as described in the third aspect of the present invention, which is used to prepare a pharmaceutical composition for preventing and / or treating diseases related to the activity or expression of MDM2.

[0054] In another preferred embodiment, the pharmaceutical composition further comprises a second therapeutic agent, and the second therapeutic agent is selected from the following group: small molecule anti-tumor drugs, antibodies, ADCs, cellular immunotherapeutics, or a combination thereof.

[0055] In another preferred embodiment, the pharmaceutical composition is used to treat a disease selected from the group consisting of bladder cancer, breast cancer, colon cancer, rectal cancer, kidney cancer, liver cancer, lung cancer (small cell lung cancer and non-small cell lung cancer), esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, prostate cancer and skin cancer (including squamous cell carcinoma); lymphoid lineage hematopoietic system tumors (including leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma); lymphoma); hematopoietic neoplasms of the myeloid lineage (including acute and chronic myeloid leukemias, myelodysplastic syndromes, and promyelocytic leukemias); tumors of mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma and other sarcomas, such as soft tissue sarcomas and osteosarcomas); tumors of the central and peripheral nervous system (including astrocytomas, neuroblastomas, gliomas, and schwannomas); and other tumors (including melanomas, seminoma, teratomas, osteosarcomas, xeroderma pigmentosum, and leukemia). igmentosum), keratoacanthoma, follicular thyroid carcinoma and Kaposi's sarcoma), endometrial cancer, head and neck cancer, glioblastoma, malignant ascites, hematopoietic cancer, thyroid hyperplasia (especially Graves' disease), cysts, asthma, chronic obstructive pulmonary disease (COPD), emphysema, psoriasis, contact dermatitis, conjunctivitis, allergic rhinitis, systemic lupus erythematosus (SLE), ulcerative colitis, Crohn's disease, multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, Alzheimer's disease, atherosclerosis, Huntington's disease, inflammatory diseases, hypoxia, ulcers, viral infections, bacterial infections, and bacterial sepsis.

[0056] In another preferred embodiment, the disease is p53 wild-type cancer.

[0057] In another preferred embodiment, the cancer is a p53 wild-type and CDKN2A mutant cancer.

[0058] In the fifth aspect of the present invention, an MDM2 inhibitor is provided, which comprises the compound according to the first aspect of the present invention, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0059] In the sixth aspect of the present invention, a method for inhibiting MDM2 activity in vitro is provided, comprising the steps of contacting the compound described in the first aspect of the present invention, or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate with MDM2 protein, thereby inhibiting MDM2 activity.

[0060] In another preferred embodiment, the method is non-therapeutic and non-diagnostic.

[0061] In the seventh aspect of the present invention, a method for treating tumors is provided, comprising the steps of administering to a subject in need thereof the compound described in the first aspect of the present invention or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition described in the third aspect of the present invention.

[0062] In another preferred embodiment, the method further comprises the step of performing concomitant diagnosis on the subject to determine whether the subject is suitable for administration of the compound or pharmaceutical composition of the present invention.

[0063] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0064] After extensive and in-depth research, the inventors discovered a class of MDM2 inhibitors with excellent inhibitory effects. Based on this, the inventors completed the present invention.

[0065] definition

[0066] As used herein, the term "alkyl" includes straight or branched chain alkyl groups. For example, C1-C8 alkyl groups represent straight or branched chain alkyl groups having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, and the like.

[0067] As used herein, the term "alkenyl" includes straight or branched alkenyl groups. For example, C2-C6 alkenyl refers to a straight or branched alkenyl group having 2 to 6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or the like.

[0068] As used herein, the term "alkynyl" includes straight or branched chain alkynyl groups. For example, C2-C6 alkynyl refers to a straight or branched chain alkynyl group having 2 to 6 carbon atoms, such as ethynyl, propynyl, butynyl, or the like.

[0069] As used herein, the term "C3-C 10 "Cycloalkyl" refers to a cyclic alkyl group having 3 to 10 carbon atoms. It may be a monocyclic ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. It may also be a bicyclic ring, such as a bridged ring or a spiro ring.

[0070] As used herein, the term "C1-C8 alkylamino" refers to an amino group substituted by a C1-C8 alkyl group, which may be monosubstituted or disubstituted; for example, methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, tert-butylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, di-tert-butylamino, etc.

[0071] As used herein, the term "C1-C8 alkoxy" refers to a straight or branched alkoxy group having 1 to 8 carbon atoms; for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, etc.

[0072] As used herein, the term "3-10 membered heterocycloalkyl group having 1-3 heteroatoms selected from the group consisting of N, S, and O" refers to a saturated or partially saturated cyclic group having 3-10 atoms, 1-3 of which are heteroatoms selected from the group consisting of N, S, and O. It may be a monocyclic or bicyclic ring, such as a bridged ring or a spirocyclic ring. Specific examples include oxetane, azetidine, tetrahydro-2H-pyranyl, piperidinyl, tetrahydrofuranyl, morpholinyl, and pyrrolidinyl.

[0073] As used herein, the term "C6-C 10 The term "aryl" refers to an aromatic group having 6 to 10 carbon atoms, for example, phenyl or naphthyl and the like.

[0074] As used herein, the term "5-10 membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S, and O" refers to a cyclic aromatic group having 5-10 atoms, 1-3 of which are heteroatoms selected from the group consisting of N, S, and O. It may be a monocyclic or condensed ring. Specific examples include pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl and (1,2,4)-triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, and oxazolyl.

[0075] Unless otherwise specified as "substituted or unsubstituted", the groups of the present invention may be substituted by substituents selected from the following groups: halogen, nitrile, nitro, hydroxyl, amino, C1-C6 alkyl-amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C2-C6 alkenyl, halogenated C2-C6 alkynyl, halogenated C1-C6 alkoxy, allyl, benzyl, C6-C 12 Aryl, C1-C6 alkoxy-C1-C6 alkyl, C1-C6 alkoxy-carbonyl, phenoxycarbonyl, C2-C6 alkynyl-carbonyl, C2-C6 alkenyl-carbonyl, C3-C6 cycloalkyl-carbonyl, C1-C6 alkyl-sulfonyl, etc.

[0076] As used herein, "halogen" or "halogen atom" refers to F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from F, Cl, and Br. "Halogenated" means substituted with an atom selected from F, Cl, Br, and I.

[0077] Unless otherwise specified, the structural formulas described herein are intended to include all isomeric forms (e.g., enantiomers, diastereomers, and geometric isomers (or conformers)): for example, R and S configurations containing asymmetric centers, (Z) and (E) isomers of double bonds, etc. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers, or geometric isomers (or conformers) are all within the scope of the present invention.

[0078] As used herein, the term "tautomer" refers to structural isomers of different energies that can interconvert across a low energy barrier. For example, proton tautomers (i.e., prototropy) include interconversion via proton migration, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversion via reorganization of some of the bonding electrons.

[0079] As used herein, the term "solvate" refers to a complex in which the compound of the present invention is coordinated with solvent molecules to form a specific ratio.

[0080] As used herein, the term "hydrate" refers to a complex formed by coordination of a compound of the present invention with water.

[0081] Active ingredient

[0082] As used herein, "compounds of the present invention" refers to compounds of formula I, and also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula I:

[0083]

[0084] in,

[0085] is a 5-membered, 6-membered or 7-membered heterocyclic group; wherein the heterocyclic group includes 1-3 N atoms and 0-2 heteroatoms selected from the group consisting of S and O;

[0086] X is C=O or S=(O)2;

[0087] n is 1, 2, 3, or 4;

[0088] Each R is independently selected from the following group: H, cyano, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C 10 Aryl, or a substituted or unsubstituted 5-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O;

[0089] Z1 is selected from the following groups: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl (including monocyclic, cyclic or bridged forms), substituted or unsubstituted C6-C 10 aryl;

[0090] Q is selected from the following group:

[0091] m and p are each independently 1, 2, 3 or 4;

[0092] Each Z2 or Z3 is independently selected from the group consisting of substituted or unsubstituted C1-C7 alkylene, NR1, O, S, C=O, S=(O)2;

[0093] Z4 is selected from Wherein, said R1 is selected from the following groups: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C 10 Aryl, cyano, -C(=O)-NRdRe, -C(=O)-substituted or unsubstituted C1-C6 alkoxy, -C(=O)-substituted or unsubstituted C1-C6 alkyl, -C(=O)-substituted or unsubstituted C3-C10 cycloalkyl, -C(=O)-substituted or unsubstituted C2-C6 alkenyl, -C(=O)-substituted or unsubstituted C2-C6 alkynyl;

[0094] Rd and Re are each independently selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C6-C 10 Aryl; or said Rd and Re form a 4-8 membered heterocyclic ring with adjacent N atoms, said heterocyclic ring containing 1-2 nitrogen atoms and 0-1 S or O atoms;

[0095] R2 is selected from the following groups: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C 10 Aryl, or a substituted or unsubstituted 5-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O;

[0096] Unless otherwise specified, the term "substituted" refers to substituted by one or more (e.g., 2, 3, 4, etc.) substituents selected from the group consisting of halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo, -CN, hydroxy, amino, carboxyl, a group selected from the group consisting of C6-C10 aryl, halogenated C6-C10 aryl, a 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, and a halogenated 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O; the substituents are selected from the group consisting of halogen, C1-C6 alkoxy;

[0097] The additional condition is that when Z4 is When Q is

[0098] As used herein, "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention formed with an acid or base that is suitable for pharmaceutical use. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts are salts of a compound of the present invention formed with an acid. Suitable acids for salt formation include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid. Suitable cations for salt formation include alkali metal and alkaline earth metal cations, such as sodium, lithium, potassium, calcium, and magnesium, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

[0099] In another preferred embodiment, the m, n, p, Z1, Z2, Z3, Z4, Q, and R are each independently a group or value corresponding to each compound in Table A or Table B.

[0100] A preferred class of compounds of the present invention is shown in Table A or Table B.

[0101] Pharmaceutical compositions and methods of administration

[0102] Since the compounds of the present invention have excellent MDM2 inhibitory activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat (stabilize, alleviate or cure) cancer. Cancers that can be treated with the compounds of the present invention include (but are not limited to) cancers such as bladder cancer, breast cancer, colon cancer, rectal cancer, kidney cancer, liver cancer, lung cancer (small cell lung cancer and non-small cell lung cancer), esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, prostate cancer and skin cancer (including squamous cell carcinoma); lymphoid lineage hematopoietic system tumors (including leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma and Burkitt's lymphoma (Burkett's Lymphoma); myeloid lineage hematopoietic tumors (including acute and chronic myeloid leukemia, myelodysplastic syndrome and promyelocytic leukemia); tumors of mesenchymal origin (including fibrosarcoma and rhabdomyosarcoma and other sarcomas, such as soft tissue sarcoma and osteosarcoma); tumors of the central and peripheral nervous systems (including astrocytomas, neuroblastomas, gliomas and schwannomas); and other tumors (including melanoma, seminoma, teratoma, osteosarcoma, xenoderomapigmentosum, keratoacanthoma, follicular thyroid carcinoma and Kaposi's sarcoma). Other cancers that can be treated with the compounds of the present invention include endometrial cancer, head and neck cancer, glioblastoma, malignant ascites, and hematopoietic cancer.

[0103] Specific cancers that may be treated with the compounds of the invention include soft tissue sarcomas, bone cancers (e.g., osteosarcoma), breast tumors, bladder cancer, Li-Fraumeni syndrome, brain tumors, rhabdomyosarcoma, adrenocortical carcinoma, colorectal cancer, non-small cell lung cancer, and acute myeloid leukemia (AML).

[0104] The pharmaceutical compositions of the present invention comprise a safe and effective amount of a compound of the present invention and a pharmaceutically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 10-200 mg per dose. Preferably, "one dose" is one capsule or tablet.

[0105] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0106] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration routes include (but are not limited to): oral, parenteral (intravenous, intramuscular or subcutaneous), and intratumoral.

[0107] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0108] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0109] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0110] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0111] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0112] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0113] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (eg, chemical anticancer drugs).

[0114] When administered in combination, the pharmaceutical composition may further comprise one or more (2, 3, 4, or more) other pharmaceutically acceptable compounds (e.g., chemical anticancer drugs). One or more (2, 3, 4, or more) of the other pharmaceutically acceptable compounds (e.g., chemical anticancer drugs) may be used simultaneously, separately, or sequentially with the compound of the present invention to treat cancer or related diseases.

[0115] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage. For a 60 kg human, the daily dosage is generally 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage will also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0116] The main advantages of the present invention include:

[0117] (1) The compounds of the present invention have novel structures and excellent MDM2 inhibitory effects. In this application, existing sulfone compounds are transformed into sulfilimine compounds, which can maintain the ability to inhibit the activity or expression of MDM2 while reducing the plasma protein binding rate, increasing the free form of the drug, and facilitating transmembrane transport to organ tissues to exert their effects.

[0118] (2) The compounds of the present invention have very low toxicity to normal cells and can therefore be used in therapeutic subjects over a wide dosage range.

[0119] (3) The compounds of the present invention have good drugability. Compared with existing compounds, the compounds of the present invention have better solubility and show good bioavailability in in vivo experiments. In addition, compared with existing compounds, the compounds of the present invention are very easy to prepare pharmaceutically acceptable salts, which helps to further form preparations.

[0120] (4) The compounds of the present invention and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat cancer-related diseases.

[0121] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0122] The various compounds appearing in the examples were prepared by the following routes:

[0123] Example 1

[0124] Synthesis of 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(propyl-2-sulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetic acid

[0125]

[0126]

[0127] Step 1: Methyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-5-hydroxy-2-methylpentanoate

[0128]

[0129] Methyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-2-methyl-5-oxopentanoate (36.5 g) was dissolved in ethanol (300 ml), cooled to 0-5°C, and NaBH4 (2.85 g) was added in batches. The reaction was allowed to proceed at 0-10°C for 2 hours. The reaction was essentially complete as monitored by TLC. Acetic acid (-8 ml) was added dropwise until no hydrogen was released. The solvent was concentrated, and 300 ml of ethyl acetate was added. The mixture was washed with water and saturated sodium bicarbonate in sequence. After drying over anhydrous magnesium sulfate, the mixture was concentrated to give 37 g of methyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-5-hydroxy-2-methylpentanoate.

[0130] Step 2: 4-(3-chlorophenyl)-5-(4-chlorophenyl)-5-hydroxy-2-methylpentanoic acid

[0131]

[0132] 37 g of methyl 4-(3-chlorophenyl)-5-(4-chlorophenyl)-5-hydroxy-2-methylpentanoate was dissolved in ethanol (300 ml), and 100 ml of an aqueous solution of LiOH.H2O (8.4 g) was added. The mixture was reacted at 20°C for 18 hours. The reaction was essentially complete as monitored by TLC. 4N hydrochloric acid was added dropwise until the pH was <1. The solvent was concentrated, and toluene was added and extracted with 250 ml of toluene at 50°C for 2 times. The mixture was washed with water to obtain a toluene solution of 4-(3-chlorophenyl)-5-(4-chlorophenyl)-5-hydroxy-2-methylpentanoic acid, which was directly used for the next step.

[0133] Step 3: 5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one

[0134]

[0135] TsOH.H2O (1.0 g) was added to a toluene solution of 4-(3-chlorophenyl)-5-(4-chlorophenyl)-5-hydroxy-2-methylpentanoic acid, and the mixture was heated to separate water and reflux for 2 hours. The reaction was basically complete as monitored by TLC. After cooling, the mixture was washed with a saturated aqueous sodium bicarbonate solution and dried over anhydrous magnesium sulfate. The toluene solution was concentrated to obtain 37.7 g of a crude product, which was separated by column chromatography to obtain 5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one.

[0136] Step 4: (±)(3S,5R,6R)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one

[0137]

[0138] Dissolve 5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one (6.7 g) and bromoacrylate (7.26 g) in THF (25 ml), cool to -50 °C, add dropwise a solution of LiHMDS (26 ml of 1 M in THF), allow to warm to 0 °C and stir for 1 h, TLC track disappearance of starting material, add saturated ammonium chloride solution, extract with ethyl acetate, column chromatography to isolate 6.0 g of product, the isomer is more difficult to purify by column, dissolve 6.0 g of product in 50 ml of n-heptane / toluene (10:1), heat to reflux to dissolve, slowly cool to room temperature, 2.8 g of solid (±)(3S,5R,6R)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2-one precipitates.

[0139] 1 HNMR (CDCI3, 400 MHz): 7.22-7.11 (m, 4H), 6.873 (d, 1H, J = 1.9 Hz), 6.745 (d, 1H, J = 7.8 Hz), 6.58-6.54 (m, 2H), 5.804 (m, 1H), 5.677 (d, 1H, J = 5.1 Hz), 5.157 (d, 1H, J = 10.2 Hz), 5.125 (dd, 1H, J = 1.6, 15.3 Hz), 3.787 (dt, 1H, J = 4.5, 12.2 Hz), 2.598 (dd, 1H, J = 7.9, 14.1 Hz), 2.488 (dd, 1H, J = 7.1, 13.7 Hz), 1.954 (t, 1H, J = 14.0 Hz), 1.897 (dd, 1H, J = 4.5, 14.0 Hz), 1.389 (s, 3H).

[0140] Step 5: 2-((2R,3R)-2-(3-chlorophenyl)-3-(4-chlorophenyl)-3-hydroxypropyl)-N-((S)-1- hydroxy-3-methylbutyl-2)-2-methylpent-4-enamide

[0141]

[0142] (3S,5R,6R)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyltetrahydro-2H-pyran-2- one (1.0 g) was dissolved in toluene (1 ml), L-valinol (0.825 g) was added and heated to 100°C for 5 hours, TLC tracking showed the reaction was almost complete, after cooling, ethyl acetate was added, washed with IN hydrochloric acid, saturated sodium bicarbonate, dried over anhydrous magnesium sulfate and concentrated to give 2-((2R,3R)-2-(3-chlorophenyl)-3-(4-chlorophenyl)-3-hydroxypropyl)-N-((S)-1- hydroxy-3-methylbutyl-2)-2-methylpent-4-enamide 1.48 g.

[0143] Step 6: trifluoromethanesulfonic acid (3S,5S,6R,8S)-8-allyl-6-(3-chlorophenyl)-5-(4- chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridine salt

[0144]

[0145] 2-((2R,3R)-2-(3-chlorophenyl)-3-(4-chlorophenyl)-3-hydroxypropyl)-N-((S)-1-hydroxy-3- methylbutyl-2)-2-methylpent-4-enamide (63.6 g) was dissolved in DCM (640 ml), 2,6-lutidine (57 g) was added, cooled to -78°C, Tf20 (97.6 g) was added dropwise, after the addition was complete, the temperature was allowed to rise to room temperature and the reaction was allowed to proceed overnight, washed with 0.5 M TfOH solution (200 ml), extracted with ethyl acetate (500 ml x 2), the organic phase was concentrated and dissolved in DCM (400 ml), column chromatography was used for separation, the conditions were:

[0146] Silica gel: 120 g

[0147] Amount loaded each time: 10 g

[0148] Mobile phase: A is n-heptane, B is acetone

[0149] Time (min) Mobile phase ratio 0-5 25％ 5-15 25％-35％ 15-30 35％ 30-35 25％

[0150] The less polar isomer (27.4 g, 34.8%) was obtained as trifluoromethanesulfonic acid (3S,5S,6R,8S)-8-allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl- 2,3,5,6,7,8-hexahydrooxazolo[3,2-a]pyridine salt:

[0151] 1HNMR(d6-DMSO,400MHz):8.15~7.10(m,8H),5.812(m,1H),5.346(dd,1H,J=1.2,16.8Hz),5.238(dd,1H,J=1.5, 10.1Hz),5.173(d,1H,J=11.3Hz),5.003(dd,1H,J=5.5,10.2Hz),4.870(t,1H,J=10.2Hz),4.323(m,1H),4.057 (ddd,1H,3.1,13.7,10.6Hz),2.812(dd,1H,J=7.1,13.7Hz),2.717(dd,1H,J=7.4,13.7Hz),2.316(t,1H,13.7H z), 1.993 (dd, 1H, J = 13.7, 3.5Hz), 1.303 (s, 3H), 0.579 (d, 3H, J = 6.7Hz), 0.524 (d, 3H, J = 7.0Hz), 0.428 (m, 1H).

[0152] The more polar isomer (22.8 g, 28.9%) is (3S,5R,6S,8R)-8-allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]-4-pyridinium trifluoromethanesulfonate:

[0153] 1 HNMR(d6-DMSO,400MHz):7.50~7.05(m,8H),5.902(m,1H),5.290(dd,1H,J=1.6,17.2Hz),5.230(dd,1 H,J=2.4,10.2Hz),5.140(d,1H,J=10.2Hz),5.084(dd,1H,J=3.9,10.2Hz),4.927(t,1H,J=10.2Hz),3. 878(m,1H),3.423(m,1H),2.733(dd,1H,J=8.3,14.1Hz),2.657(dd,1H,J=6.7,13.7Hz),2.334(t,1H, 13.7Hz), 2.005 (dd, 1H, J = 13.7, 2.7Hz), 1.334 (s, 3H), 0.884 (d, 3H, J = 6.6Hz), 0.662 (d, 3H, J = 6.6Hz).

[0154] Step 7: (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylmercapto)-3-methylbutyl-2-yl)-3-methylpiperidin-2-one

[0155]

[0156] Under nitrogen protection, isopropyl mercaptan (10.4 g) was added dropwise to potassium tert-butoxide (82.2 ml 1M The mixture was added into a DMF (THF) solution, cooled appropriately so that the temperature did not exceed 30°C, stirred for 10 minutes after the addition, and a DMF solution (80 ml) of (3S,5S,6R,8S)-8-allyl-6-(3-chlorophenyl)-5-(4-chlorophenyl)-3-isopropyl-8-methyl-2,3,5,6,7,8-hexahydrooxazolo[3,2-a]-4-pyridinium trifluoromethanesulfonate (17.8 g) was added. The mixture was heated to 50°C for 3 hours. After completion of the reaction by TLC, the mixture was poured into 600 ml of water, extracted with ethyl acetate (200 ml×3), washed with water, concentrated, and separated by column chromatography to obtain 13.5 g of an oily product (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylmercapto)-3-methylbutyl-2)-3-methylpiperidin-2-one.

[0157] Step 8: (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfoxide)-3-methylbutyl-2-methylpiperidin-2-one

[0158]

[0159] (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylmercapto)-3-methylbutyl-2-yl)-3-methylpiperidin-2-one (2.03 g) was dissolved in methanol (25 ml), 0.60 ml of 30% hydrogen peroxide was added, and the mixture was reacted at 20-25° C. for 2 days. Sodium thiosulfate solution was added to terminate the reaction, and the mixture was extracted with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and concentrated. Column chromatography was used to separate 1.99 g of (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfoxide)-3-methylbutyl-2-yl)-3-methylpiperidin-2-one, which was a mixture of two isomers in a yield of 95%.

[0160] 1HNMR (CDCl3, 400MHz): 7.27~7.00 (m, 7H), 6.978 (d, 1H, J = 6.8Hz), 5.898 (m, 1H), 5.260 (d, 1H, J = 16.8Hz), 5.180 (dd, 1H, J = 1.2, 10.0Hz), 4.86 0(d,1H,J=8.4Hz),3.620(t,1H,J=10.5Hz),3.541(ddd,1H,J=2.3,11.0,13.3Hz),3.017(m,2H),2.860(dd,1H,J=2.7,12.9Hz),2.741(dd,1H, J=8.2,14.1Hz),2.716(dd,1H,J=2.8,13.3Hz),2.585(dd,1H,J=6.7,13.7Hz),2.188(m,1H),2.101(t,1H,J=13.7Hz),1.922(dd,1H,J=2.8,1 3.7Hz), 1.343 (d, 3H, J = 7.0Hz), 1.277 (d, 3H, J = 7.0Hz), 1.199 (s, 3H), 0.863 (t, 1H, J = 6.6Hz), 0.675 (d, 3H, J = 6.7Hz), 0.481 (d, 3H, J = 7.0Hz).

[0161] Step 9: 2,2,2-Trifluoroacetyl N-(((S)-2-((3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-2-oxopiperidin-1-yl)-3-methylbutyl)(isopropyl)(oxy)-16-sulfoneguanylamine

[0162]

[0163] (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-l-((S)-l-(isopropylsulfinyl)-3- methylbutyl-2)-3-methylpiperidin-2-one (336 mg) and trifluoroacetamide (142.4 mg) were dissolved in DCM (4 ml), then MgO (126.5 mg) and Rh2(OAc)4(6.9 mg) were added, and finally PhI(OAc)2(303.7 mg) was added. The mixture was stirred at room temperature overnight, filtered, and the solid was washed with DCM. The solution was concentrated and passed through a column, eluting with n-heptane / ethyl acetate = 2:1, to give 58 mg of the product 2,2,2-trifluoroacetamidoN-(((S)-2-((3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-2-oxopiperidin-l-yl)-3-methylbutyl)(isopropyl)(oxido)-l6-sulfamidylamine) as a mixture of two isomers in 14% yield.

[0164] 1 HNMR (CDCI3, 400 MHz): 7.3-7.1 (m, 6H), 6.932 (s, 1H), 6.860 (t, 1H, J = 3.2 Hz), 5.904 (m, 1H), 5.239 (m, 2H), 4.959 (d, 1H, J = 10.8 Hz), 4.450 (dd, 1H, J = 7.6, 15.2 Hz), 4.135 (m, 1H), 3.368 (m, 2H), 3.186 (ddd, 1H, J = 1.6, 7.6, 9.2 Hz), 2.658 (m, 2H), 2.390 (m, 1H), 2.229 (t, 1H, J = 13.6 Hz), 1.898 (dd, 1H, J = 3.2, 14.0 Hz), 1.522 (d, 3H, J = 7.0 Hz), 1.484 (d, 3H, J = 7.0 Hz), 1.282 (t, 1H, J = 7.2 Hz), 1.256 (s, 3H), 0.684 (d, 3H, J = 6.8 Hz), 0.620 (d, 3H, J = 7.2 Hz).

[0165] Step 10: 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-l-((2S)-3-methyl-l-(N-(2,2,2- trifluoroacetyl)-2-propylsulfamidyl)butyl-2-)-2-oxopiperidin-3-yl)acetic acid

[0166]

[0167] 2,2,2-Trifluoroacetyl N-(((S)-2-((3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-2-oxopiperidin-1-yl)-3-methylbutyl)(isopropyl)(oxy)-16-sulfoneguanylamine) (150 mg) and ruthenium trichloride (7.3 mg) were dissolved in DCM / water (5 ml / 5 ml), and then tetrabutylammonium hydrogen sulfate (15.8 mg) and sodium periodate (300 mg) were added and the mixture was stirred at room temperature. The mixture was stirred overnight, filtered, and extracted with DCM. The solution was dried and concentrated, and then passed through a column with n-heptane / ethyl acetate = 1:1 as the eluent to obtain 120 mg of the product 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-(2,2,2-trifluoroacetyl)-2-propylsulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetic acid, which was a mixture of two isomers with a yield of 77.6%.

[0168] 1 H NMR(400MHz,CD3OD)δ7.27(br,4H),7.19–6.89(m,4H),5.00(dd,1H,J=20.4,11.0Hz ),4.44(ddd,1H,J=28.6,15.1,9.2Hz),4.08(m,1H),3.71–3.48(m,2H),3.39–3.17( m,2H),3.05–2.92(m,1H),2.67–2.55(m,1H),2.34–1.93(m,4H),1.51(d,3H,J=7.1H z), 1.47 (d, 3H, J = 7.1Hz), 1.46 (s, 3H), 1.35 (d, 3H, J = 3.2Hz), 0.47 (d, 3H, J = 7.0Hz).

[0169] Step 11: 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(2-propylsulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetic acid

[0170]

[0171] (2S)-3-methyl-l-(N-(2,2,2-trifluoroacetyl)-2-propylsulfonamidyl)butyl-2-)-2-oxopiperidin-3-yl)acetic acid (100 mg) was dissolved in methanol (2.0 ml), then potassium carbonate (417 mg) was added and stirred at room temperature for 2 hours, water and ethyl acetate were added, the pH was adjusted to 6 with 3N HCI, the solution was dried and concentrated, then column chromatography was performed, the eluent was n-heptane / ethyl acetate = 1 : 1 to 0: 1, to obtain two isomer products, 31 mg of the more polar and 18 mg of the less polar, total yield 57%.

[0172] More polar compound 001: 1 H NMR (400 MHz, CDC13) δ 7.22 (br, 4H), 7.10 - 6.91 (m, 3H), 6.84 (dd, 1H, J = 5.4, 3.3 Hz), 5.38 (d, 1H, J = 11.0 Hz), 4.12 - 3.97 (m, 1H), 3.40 - 3.20 (m, 2H), 3.13 (m, 1H), 2.94 - 2.72 (m, 2H), 2.37 (t, 1H, J = 13.7 Hz), 2.19 (m, 1H), 1.91 (dd, 1H, J = 13.8, 3.0 Hz), 1.57 (d, 1H, J = 6.8 Hz), 1.41 (s, 3H), 1.37 (d, 3H, J = 7.1 Hz), 1.35 (d, 3H, J = 7.1 Hz), 1.30 - 1.13 (m, 3H), 0.61 (d, 2H, J = 6.6 Hz), 0.44 (d, 2H, J = 6.8 Hz).

[0173] Less polar compound 002: 1 H NMR (400 MHz, CDC13) δ 7.22 (br, 4H), 7.10 - 6.91 (m, 3H), 6.84 (dd, 1H, J = 5.4, 3.3 Hz), 5.38 (d, 1H, J = 11.0 Hz), 4.12 - 3.97 (m, 1H), 3.40 - 3.20 (m, 2H), 3.13 (m, 1H), 2.94 - 2.72 (m, 2H), 2.37 (t, 1H, J = 13.7 Hz), 2.19 (m, 1H), 1.91 (dd, 1H, J = 13.8, 3.0 Hz), 1.57 (d, 1H, J = 6.8 Hz), 1.41 (s, 3H), 1.37 (d, 3H, J = 7.1 Hz), 1.35 (d, 3H, J = 7.1 Hz), 1.30 - 1.13 (m, 3H), 0.61 (d, 2H, J = 6.6 Hz), 0.44 (d, 2H, J = 6.8 Hz).

[0174] Example 2

[0175] Synthesis of 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-methylpropyl-2-sulfoneamidino)butyl-2-)-2-oxopiperidin-3-yl)acetic acid

[0176]

[0177] Step 1: (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(2-propylsulfonamidino)butyl-2-)piperidin-2-one

[0178]

[0179] 2,2,2-Trifluoroacetyl N-(((S)-2-((3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-2-oxopiperidin-1-yl)-3-methylbutyl)(isopropyl)(oxy)-16-sulfoneguanylamine) (512 mg) was dissolved in methanol (15 ml), and potassium carbonate (600 mg) was added and stirred at room temperature overnight. Water and ethyl acetate were separated, the solution was dried and concentrated, and then passed through a column with the eluent being n-heptane / ethyl acetate = 1:1 to 0:1 to obtain 352 mg of a mixture of two isomers (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(2-propylsulfonamidino)butyl-2-)piperidin-2-one with a total yield of 80%.

[0180] 1H NMR (400MHz, CDCl3) δ7.27(br,4H),7.16–6.97(m,3H),6.97–6.83(m,1H),5.90(ddt,1H,J=17.3,10.1,7 .4Hz),5.36(d,1H,J=10.9Hz),5.31–5.15(m,2H),4.25–4.01(m,1H),3.37(ddt,2H,J=13.8,10.3,6.1Hz ),3.28–3.06(m,1H),3.04–2.84(m,1H),2.66(m,2H),2.26(m,2H),1.82(dd,1H,J=13.5,3.2Hz),1.67(s ,2H),1.40(d,J=7.0,3H),1.37(d,J=7.0,3H),1.22(s,3H),0.66(d,3H,J=6.7Hz),0.53(d,3H,J=6.8Hz).

[0181] Step 2: (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-methyl-2-propylsulfonamidino)butyl-2-)piperidin-2-one

[0182]

[0183] (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(2-propylsulfonamidino)butyl-2-)piperidin-2-one (350 mg) was dissolved in DMF (7.0 ml), cooled to 0°C, 60% sodium hydride (38 mg) was added, and the mixture was stirred for 15 min. Methyl iodide (180 mg) was then added, and the temperature was raised to room temperature for overnight reaction. The mixture was poured into ice water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The mixture was separated by column chromatography to obtain (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-methyl-2-propylsulfonamidino)butyl-2-)piperidin-2-one.

[0184] Step 3: Synthesis of 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-methyl-2-propylsulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetic acid

[0185]

[0186] (3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-methyl-2-propylsulfonamidino)butyl-2-)piperidin-2-one (141.6 mg) and ruthenium trichloride (7.8 mg) were dissolved in acetonitrile / water (5 ml / 5 ml), and then sodium periodate (322 mg) was added and stirred at room temperature overnight, filtered, and extracted with DCM. After the solution was dried and concentrated, it was passed through a column with the eluent being n-heptane / ethyl acetate = 1:1 to obtain two isomers of 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-methyl-2-propylsulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetic acid, of which the less polar isomer was 003, yielding 47.5 mg, with a yield of 32%.

[0187] 1 H NMR (400MHz, CDCl3) δ7.59–7.19(m,3H),7.18–7.02(m,4H),6.85(d,J=7.4Hz,1H),5.24(d,J=10.9H z,1H),3.90(dd,J=13.5,10.6Hz,1H),3.51–3.24(m,4H),3.03(d,J=15.0Hz,1H),2.98(s,3H),2.88 –2.75(m,1H),2.57(t,J=13.8Hz,1H),2.20(dq,J=13.6,7.2,6.6Hz,2H),2.00–1.83(m,2H),1.57–1 .40(m,10H),1.35–1.25(m,3H),0.90(t,J=6.7Hz,1H),0.66(d,J=6.2Hz,3H),0.45(d,J=6.9Hz,2H).

[0188] The more polar isomer was 004, yielding 40 mg, with a yield of 28%.

[0189] 1H NMR (400MHz, CDCl3) δ7.59–7.19(m,3H),7.15–6.93(m,5H),6.83(d,J=7.3Hz,1H),5.31(d,J=11. 0Hz,1H),3.97(m,1H),3.42(m,1H),3.28(m,3H),3.04(d,J=14.7Hz,1H),2.97(s,3H),2.80(d,J= 15.5Hz,2H),2.32(t,J=13.7Hz,1H),2.26–2.16(m,1H),1.91(t,J=14.5Hz,1H),1.48(d,J=6.8Hz ,3H),1.43(d,J=6.8Hz,3H),0.90(t,J=6.7Hz,1H),0.66(d,J=6.6Hz,3H),0.50(d,J=7.0Hz,3H).

[0190] Example 3: Synthesis of 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((2S)-1-(N-cyanopropyl-2-sulfoneamidino)-3-methylbutyl-2-)-3-methyl-2-oxopiperidinyl-3-)acetic acid

[0191]

[0192] Step 1: Synthesis of N-(((S)-2-((3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-2-oxopiperidin-1-yl)-3-methylbutyl)(isopropyl)(oxy)-16-sulfanylidene)cyanamide

[0193]

[0194] Referring to Step 9 in Example 1, trifluoroacetamide was replaced with cyanamide to obtain N-(((S)-2-((3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-2-oxopiperidin-1-yl)-3-methylbutyl)(isopropyl)(oxy)-16-sulfanylidene)cyanamide.

[0195] Step 2: Synthesis of 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((2S)-1-(N-cyanopropyl-2-sulfone amidino)-3-methylbutyl-2-)-3-methyl-2-oxopiperidinyl-3-)acetic acid

[0196]

[0197] Referring to Step 10 in Example 1, 2,2,2-trifluoroacetyl N-(((S)-2-((3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-2-oxopiperidin-1-yl)-3-methylbutyl)(isopropyl)(oxy)-16-sulfoneguanylamine was replaced with N-(((S)-2-((3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-2-oxopiperidin-1-yl)-3-methylbutyl)(isopropyl)(oxy)-16-sulfoneguanylamine A pair of isomers of 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((2S)-1-(N-cyanopropyl-2-sulfone amidino)-3-methylbutyl-2-)-3-methyl-2-oxopiperidinyl-3-)acetic acid were obtained by preparative HPLC separation. Peak 1 isomer is 005: 1 H NMR (400MHz, CDCl3) δ7.59-7.19(m,3H),7.19–7.04(m,3H),7.04–6.94(m,2H),4.98(d,J=10.3Hz,1H),4.40( dd,J=12.5Hz,1H),3.68(m,1H),3.47(t,J=8.9Hz,1H),3.28(t,J=12.0Hz,1H),2.93(dd,J=13.9,2.0Hz,1H), 2.89(m,3H),2.21(m,1H),2.12(q,J=6.9Hz,1H),2.03(d,J=13.4Hz,1H),1.63(d,J=6.2Hz,3H),1.61(d,J=6. 2Hz, 3H), 1.49 (s, 3H), 0.90 (t, J = 6.8Hz, 2H), 0.66 (d, J = 6.6Hz, 3H), 0.54 (d, J = 6.9Hz, 3H). LC-MS: M+1 = 592.2

[0198] Peak 2 Isomer 006: 1H NMR (400MHz, CDCl3) δ7.59-7.19(m,3H),7.19–7.04(m,3H),7.04–6.94(m,2H),5.01(d,J=10.3Hz,1H),4.40(dd, J=12.5Hz,1H),3.66(tt,J=13.7,6.8Hz,1H),3.47(t,J=8.9Hz,1H),3.38(t,J=12.0Hz,1H),3.00(dd,J=13.9,2.0 Hz,1H),2.89(m,3H),2.35(m,1H),2.12(q,J=6.9Hz,1H),2.03(d,J=13.4Hz,1H),1.63(d,J=6.2Hz,3H),1.61(d,J =6.2Hz, 3H), 1.44 (s, 3H), 0.90 (t, J = 6.8Hz, 2H), 0.70 (d, J = 6.6Hz, 3H), 0.50 (d, J = 6.9Hz, 3H). LC-MS: M+1 = 592.2.

[0199] Example 4: Preparation of Compound 007

[0200]

[0201] Step 1: Synthesis of methyl 4-(2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-(2,2,2-trifluoroacetyl)-2-propylsulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetamido)benzoate

[0202]

[0203] Under nitrogen, 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-(2,2,2-trifluoroacetyl)-2-propylsulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetic acid (800 mg), methyl p-aminobenzoate (219 mg), DMAP (324 mg), and EDC.HCl (508.4 mg) were added sequentially to a DCM (16 ml) solution at 0°C. The mixture was stirred at room temperature overnight. The reaction was complete as determined by TLC. Water was added dropwise to the reaction solution under ice bath to quench the reaction. The pH was adjusted to 2 with cold 1N HCl solution. The layers were separated, and the organic phase was washed once more with 1N HCl, washed with water, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was separated by column chromatography to obtain 600 mg of a white foamy solid (yield: 62.6%). 1H NMR (400MHz, CDCl3) δ8.84(s,1H),8.07(d,J=8.7Hz,2H),7.71(d,J=8.7Hz,2H),7.27–7.06(m,6H),6.97(s ,1H),6.91(t,J=3.6Hz,1H),4.97(d,J=10.9Hz,1H),4.56(dd,J=13.9,10.6Hz,1H),4.14(q,J=7.2Hz,1H),3 .94(s,3H),3.39(dd,J=16.4,11.7Hz,3H),2.89(q,J=14.4Hz,2H),2.38(t,J=13.8Hz,1H),2.15–1.92(m,2 H),1.58(d,J=6.8Hz,3H),1.53(d,J=6.9Hz,3H),1.46(s,3H),0.75(d,J=6.7Hz,3H),0.36(d,J=7.0Hz,3H).

[0204] Step 2: Synthesis of 4-(2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(propylsulfonamidino-2-)butyl-2-)-2-oxopiperidin-3-yl)acetamido)benzoic acid

[0205]

[0206] Methyl 4-(2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-(2,2,2-trifluoroacetyl)-2-propylsulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetamido)benzoate (600 mg) and LiOH.H₂O (127 mg) were added sequentially to a solution of MeOH / H₂O / THF (2.4 ml / 1.2 ml / 1.2 ml) at room temperature. The mixture was stirred at room temperature after addition. The reaction was complete as determined by TLC. The solvent was concentrated, and 10 ml of water and 20 ml of ethyl acetate were added to the residue. The pH was adjusted to 2 with cold 1N HCl solution. The layers were separated, and the aqueous layer was extracted once more with ethyl acetate. The organic phases were combined, washed with water and saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was preparatively isolated and lyophilized to obtain 250 mg of a white solid with a yield of 48.4%. 1H NMR (400MHz, CD3OD) δ8.05(d,J=7.5Hz,2H),7.81(d,J=8.7Hz,2H),7.20(m,6H),6.86(d,J=19.2Hz,2H),5.13(s,1H),5.01(s,1H),4.43(s,1H),3. 57(m,3H),3.08(d,J=13.6Hz,1H),2.67(d,J=13.9Hz,1H),2.48–2.06(m, 3H),1.47(d,J=38.1Hz,9H),0.77(s,3H),0.59(s,3H).LC-MS:M+1=686.2

[0207] Example 5:

[0208] By using different anilines or sulfonamides in place of the methyl group of p-aminobenzoic acid and the synthetic method of Example 4, the corresponding compounds 008 to 024 (see table) were obtained.

[0209] Example 6:

[0210]

[0211] Step 1:

[0212]

[0213] A toluene solution of TBS-Cl (22.5 g) in 75 ml of toluene was added dropwise to a solution of cyclopropylsulfonamide (15 g) and triethylamine (19.3 g) in THF (240 ml) at room temperature under nitrogen. The mixture was stirred at room temperature for 48 hours. MTBE (150 ml) was added to the reaction solution, stirred at room temperature for 30 minutes, and the solid was filtered off and washed with MTBE. The filtrate was mixed with silica gel and separated by column chromatography to obtain 17.8 g of the product as a white solid with y=52%. 1H NMR (400MHz, CDCl3) δ4.28(s,1H),2.61–2.35(m,1H),1.22–1.10(m,2H),1.03–0.99(m,2H),0.97(s,9H),0.31(s,6H).

[0214] Step 2:

[0215]

[0216] Under nitrogen protection at room temperature, PPh3 (13.4g) and C2Cl6 (12.1g) were added to a reaction flask. The solution was vacuum-purged three times, and then redistilled chloroform (100ml) was added. The solution was heated to 50°C and kept warm for 2h. It was then cooled to 0°C and anhydrous TEA (17.2g) was added. The mixture was stirred for 30min after addition. Then, a solution of N-tert-butyldimethylsilylcyclopropylsulfonamide (10g) in chloroform (40ml) was added. The mixture was stirred for 30min after addition. Finally, anhydrous ammonia was passed through the reaction solution (about 20min) to precipitate a large amount of solid. TLC analysis showed that the reaction was almost complete. The reaction solution was filtered to remove the solid, washed with MTBE, and the filtrate was mixed with silica gel and separated by column chromatography. The product was collected and concentrated to obtain 9.4g of a white solid with y=94%. 1 H NMR (400MHz, CDCl3) δ4.50 (s, 2H), 2.59 (ddd, J = 12.6, 7.8, 4.7Hz, 1H), 1.16–1.08 (m, 1H), 1. 02(ddd,J=15.7,6.6,4.2Hz,1H),0.96–0.91(m,2H),0.89(s,9H),0.11(s,3H),0.11(s,3H).

[0217] Step 3:

[0218]

[0219] Under nitrogen in an ice bath, add TFA (5 ml) dropwise to a solution of N'-(tert-butyldimethylsilyl)cyclopropylsulfoneguanamide (8.5 g) in DCM (15 ml). Stir at room temperature for 2 h. TLC indicates the reaction is nearly complete. Concentrate the reaction mixture to dryness, add toluene, and reconcentrate to dryness. Slurry the mixture with MTBE, filter, and vacuum dry to obtain a white solid (7.5 g, y = 80%). 1 H NMR (400MHz, DMSO-d6) δ8.74 (s, 3H), 3.07 (tt, J = 7.8, 3.9Hz, 1H), 1.33–1.12 (m, 4H).

[0220] Step 4:

[0221]

[0222] According to the method of step 10 in Example 1, 2,2,2-trifluoroacetyl N-(((S)-2-((3S,5R,6S)-3-allyl-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-2-oxopiperidin-1-yl)-3-methylbutyl)(isopropyl)(oxy)-16-sulfone amidinoamine) was replaced with (3S,5R,6S)-3-allyl-5-( 3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutyl-2-)-3-methylpiperidin-2-one to give 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutyl-2-)-3-methyl-2-oxopiperidinyl-3-)acetic acid. 1 H NMR (400MHz, CD3OD) δ7.30 (s, 3H), 7.18–7.07 (m, 4H), 7.03 (dt, J = 7.5, 1.5Hz, 1H), 5.13 (d, J = 11.0Hz, 1H),4.02(dd,J=13.9,10.4Hz,1H),3.63(ddd,J=13.9,11.1,3.1Hz,1H),3.12(dd,J=13.9,2.1Hz,1H) ,2.90(d,J=13.4Hz,1H),2.50(d,J=13.3Hz,1H),2.28(t,J=13.6Hz,1H),2.25–2.17(m,1H),2.11(dd, J=13.6,3.2Hz,1H),1.43(d,J=6.8Hz,6H),1.37(s,3H),0.68(d,J=6.6Hz,3H),0.54(d,J=6.9Hz,3H).

[0223] Step 5:

[0224]

[0225] Under nitrogen protection in an ice bath, 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutyl-2-)-3-methyl-2-oxopiperidinyl-3-acetic acid (800 mg), cyclopropylsulfonamidinesulfonamide trifluoroacetate (705.3 mg), DMAP (736.4 mg), and EDC-HCl (508.4 mg) were added sequentially to DCM (20 ml). The mixture was stirred at room temperature for 18 hours. HPLC analysis revealed no residual starting material. The reaction solution was washed with 1N HCl, separated, and the aqueous layer extracted once with DCM. The organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated to give 940 mg of a crude foamy product. A portion was purified by preparative HPLC to give 28.6 mg of the desired product with an HPLC purity of 97.6%. 1 H NMR(400MHz,CD3OD)δ7.29(br,4H),7.18–7.05(m,4H),5.13(d,J=11.1Hz,1H),4.09–3.9 7(m,1H),3.78–3.62(m,1H),3.30(m,1H),3.19–3.07(m,1H),3.00(dd,J=12.8,2.1Hz,1H ),2.97–2.91(m,1H),2.62–2.49(m,1H),2.33–2.08(m,3H),1.43(d,J=6.8Hz,6H),1.40( s,3H),1.29–0.95(m,5H),0.67(d,J=6.6Hz,3H),0.53(d,J=6.9Hz,3H).LC-MS:M+1=670.2

[0226] Example 7:

[0227] Step 1:

[0228]

[0229] Under nitrogen at room temperature, a toluene solution of TBS-Cl (6.05 g) in 30 ml of toluene was added dropwise to a solution of 1-methylcyclopropylsulfonamide (4.5 g) and TEA (6.74 g) in THF (80 ml). The mixture was stirred at room temperature for 48 hours. TLC indicated that the reaction was nearly complete. MTBE (150 ml) was added to the reaction solution and stirred at room temperature for 30 minutes. The solid was filtered and washed with MTBE. The filtrate was mixed with silica gel and separated by column chromatography to yield 2.9 g of the product as a white solid with y=43.9%. 1H NMR (400MHz, CDCl3) δ4.08 (br, 1H), 1.55 (s, 3H), 1.38 (t, J = 3.0Hz, 2H), 0.97 (s, 9H), 0.80–0.74 (m, 2H), 0.29 (s, 6H).

[0230] Step 2:

[0231]

[0232] Under nitrogen protection at room temperature, PPh3 (3.65g) and C2Cl6 (3.3g) were added to a reaction flask. The mixture was vacuum-purged three times, and then redistilled chloroform (70ml) was added. The solution was heated to 50°C and kept warm for 2h. It was then cooled to 0°C and anhydrous TEA (4.7g) was added. The mixture was stirred for 30min after addition. Then, a solution of N-tert-butyldimethylsilyl 1-methylcyclopropanesulfonamide (2.9g) in chloroform (40ml) was added. The mixture was stirred for 30min after addition. Finally, anhydrous ammonia gas was passed through the reaction solution (about 20min), resulting in the precipitation of a large amount of solid. TLC analysis showed that the reaction was essentially complete. The reaction solution was filtered to remove the solid, washed with MTBE, and the filtrate was mixed with silica gel and separated by column chromatography. The product was collected and concentrated to obtain 2.3g of a white solid with y = 79.3%.

[0233] Step 3:

[0234]

[0235] Under nitrogen in an ice bath, TFA (5 ml) was added dropwise to a solution of N'-(tert-butyldimethylsilyl)-1-methylcyclopropylsulfone amidamide (2.3 g) in DCM (15 ml). The mixture was stirred at room temperature for 2 h. TLC indicated the reaction was nearly complete. The reaction solution was concentrated to dryness, then toluene was added and concentrated again to dryness. MTBE was added to the slurry, and the white solid was filtered and dried under vacuum to yield 1.87 g (y = 81.3%).

[0236] Step 4:

[0237]

[0238] Referring to step 5 in Example 6, a pair of isomers were obtained.

[0239] 028A: 1H NMR(400MHz,CD3OD)δ7.28(br,3H),7.17–7.06(m,5H),5.11(d,J=11.1Hz,1H),4.03(dd,J=13.7,10.5Hz,1H),3.73–3.64(m,1H),3.30(d,J=7.3Hz,2H),3.12(d,J=13.8Hz,1H),3.04(d,J=12.9Hz,1H),2.54(d,J=12.9Hz,1H),2.27(t,J=13.4Hz,1H),2.23–2.15(m,2H),1.55(s,4H),1.43(d,J=6.9Hz,6H),1.39(s,3H),1.00–0.84(m,3H),0.68(d,J=6.6Hz,3H),0.52(d,J=6.9Hz,3H).LC-MS:M+1=684.2

[0240] 028B: 1 H NMR(400MHz,CD3OD)δ7.23(br,4H),7.16–7.12(m,2H),7.09(dd,J=6.0,4.5Hz,2H),5.11(d,J=11.1Hz,1H),4.03(dd,J=13.7,10.6Hz,1H),3.67(t,J=10.7Hz,1H),3.30(d,J=4.0Hz,2H),3.11(d,J=13.7Hz,1H),3.01(d,J=12.8Hz,1H),2.52(d,J=12.8Hz,1H),2.30(d,J=13.5Hz,1H),2.26–2.20(m,1H),2.16(dd,J=13.5,3.2Hz,1H),1.54(s,3H),1.53–1.48(m,1H),1.43(d,J=6.8Hz,7H),1.38(s,3H),0.97–0.83(m,3H),0.68(d,J=6.6Hz,3H),0.54(d,J=6.9Hz,3H).LC-MS:M+1=684.2

[0241] 实施例8:

[0242] 合成路线:

[0243]

[0244] 步骤1:

[0245]

[0246] Under nitrogen in an ice bath, 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutyl-2-)-3-methyl-2-oxopiperidinyl-3-acetic acid (600 mg), p-toluenesulfonamide (271 mg), DMAP (284 mg), and EDC-HCl (445 mg) were added sequentially to DCM (12 ml). The mixture was stirred at room temperature for 18 hours. HPLC analysis indicated no residual starting material. The reaction mixture was washed with 1N HCl, separated, and the aqueous layer extracted once with DCM. The organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, concentrated, and purified by column chromatography to afford 565 mg of the product as a white foamy solid, y = 74.3%. 1 HNMR(400MHz, CDCl3)δ10.93(s,1H),8.00(d,J=8.2Hz,2H),7.40(br,1H),7.35(d,J=8.2Hz,2H),7.27–7.12(m,3H), 7.09(d,J=4.7Hz,2H),6.97(s,1H),6.93–6.84(m,1H),5.12(d,J=11.0Hz,1H),4.09(dd,J=13.6,10.7Hz,1H),3.35(t ,J=9.2Hz,1H),3.26–3.11(m,2H),2.88(t,J=13.4Hz,2H),2.62(d,J=15.0Hz,1H),2.46(s,3H),2.39(s,1H),2.31–2 .21(m,1H),1.87(d,J=13.1Hz,1H),1.47(d,J=6.9Hz,6H),1.31(s,3H),0.71(d,J=6.6Hz,3H),0.49(d,J=6.9Hz,3H).

[0247] Step 2:

[0248]

[0249] PPh3 (250 mg) and C2Cl6 (223 mg) were added to a reaction flask. The atmosphere was replaced with oil / N2, and chloroform (10 ml) was added to dissolve the mixture. The mixture was heated to approximately 50°C and maintained for 2 h, resulting in the precipitation of a large amount of white solid. The solution was cooled to 0°C, and anhydrous TEA (284 mg) was added and maintained for 30 min. A solution of 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-1-((S)-1-(isopropylsulfonyl)-3-methylbutyl-2-)-3-methyl-2-oxopiperidinyl-3-)-N-p-toluenesulfonamide (565 mg) in CHCl3 (10 ml) was then added. The mixture was maintained for 30 min, and ammonia gas was introduced for 20 min. LCMS indicated the presence of the desired product. The reaction mixture was diluted with DCM, washed with water, dried over anhydrous magnesium sulfate, concentrated, and purified by HPLC to yield a solid. 1 H NMR (400MHz, CD3OD) δ7.74(br,1H),7.60(d,J=8.3Hz,2H),7.33(br,3H),7.15(d,J=6.5Hz,2H),7.09(d,J=8.0Hz,2H),7.01( s,1H),6.94(d,J=7.3Hz,1H),5.03(d,J=11.2Hz,1H),4.04–3.93(m,1H),3.63–3.52(m,1H),3.31–3.25(m,2H),3.09(dd,J=1 3.7,1.8Hz,1H),3.00(d,J=13.0Hz,1H),2.52(d,J=12.9Hz,1H),2.33(s,3H),2.18–2.08(m,1H),2.01(t,J=13.5Hz,1H),1.7 4(dd,J=13.7,2.8Hz,1H),1.41(d,J=6.9Hz,6H),1.23(s,3H),0.65(d,J=6.6Hz,3H),0.46(d,J=6.9Hz,3H).LC-MS:M+1=720.2

[0250] Example 9:

[0251] Synthesis route:

[0252]

[0253] Step 1:

[0254]

[0255] Under nitrogen at room temperature, 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-(2,2,2-trifluoroacetyl)-2-propylsulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetic acid (800 mg) was dissolved in acetone (10 ml). Anhydrous potassium carbonate (333 mg) was added, and finally iodomethane (342 mg) was added dropwise. The mixture was stirred at room temperature for 5-6 hours. TLC indicated that the reaction was complete. The solid was filtered, washed with ethyl acetate, and the filtrate was concentrated. The residue was extracted with ethyl acetate / water. The aqueous layer was re-extracted with ethyl acetate. The combined ethyl acetate layers were washed with saturated brine, dried over anhydrous magnesium sulfate, and purified by column chromatography to yield 775 mg of a white solid.

[0256] Step 2:

[0257]

[0258] Under nitrogen, methyl 2-((3R,5R,6S)-5-(3-chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((2S)-3-methyl-1-(N-(2,2,2-trifluoroacetyl)-2-propylsulfonamidino)butyl-2-)-2-oxopiperidin-3-yl)acetate (775 mg) was dissolved in anhydrous methanol (10 ml). 30% sodium methoxide / methanol (5 ml) was added dropwise under ice-cooling. The mixture was slowly warmed to room temperature with stirring. TLC confirmed the reaction was complete. Acetic acid was added dropwise to pH 6 under ice-cooling. The methanol was concentrated to remove the residue, and the residue was extracted with ethyl acetate / water. The aqueous layer was re-extracted with ethyl acetate. The ethyl acetate layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and purified by column chromatography to obtain 556 mg of a white solid in a yield of 83.4%.

[0259] Steps 3 and 4:

[0260]

[0261] Compound 19 (200 mg) was dissolved in DCM (5 ml) under nitrogen protection. Sodium hydroxide (60%, 20.6 mg) was added under ice-salt bath, and the mixture was stirred for 10 min. Acetyl chloride (30 mg) was then added, and the mixture was slowly warmed to room temperature and stirred for 2 hours. Lithium hydroxide monohydrate (42.8 mg) / water / methanol (20 ml each) were added at 0°C, and the mixture was stirred at room temperature for approximately 2 h. 2N hydrochloric acid was added until the pH was approximately 4. The methanol was concentrated under reduced pressure, and the residue was extracted with ethyl acetate / water. The aqueous layer was re-extracted with ethyl acetate. The ethyl acetate layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, concentrated, and separated and purified by preparative HPLC to obtain a white solid. 1H NMR(400MHz,CD3OD)δ7.51–6.97(m,8H),5.13(d,J=11.0Hz,1H),4.35(dd,J=14.1,10.4Hz,1H), 4.08(p,J=7.0Hz,1H),3.61–3.52(m,2H),3.32–3.29(m,1H),3.00(d,J=12.9Hz,1H),2.63(d,J= 12.2Hz,1H),2.30(t,J=13.7Hz,1H),2.19–2.05(m,2H),2.13(s,3H),1.51(d,J=6.8Hz,3H),1.4 6(d,J=6.9Hz,3H),1.39(s,3H),0.67(d,J=6.6Hz,3H),0.51(d,J=6.9Hz,3H).LC-MS: M+1=609.2

[0262] Example 10:

[0263] With reference to Example 9, 041 can be obtained by replacing acetyl chloride with acryloyl chloride; 045 can be obtained by replacing acetyl chloride with dimethylaminoformyl chloride; 046 can be obtained by replacing acetyl chloride with 4-morpholinecarbonyl chloride; 047 can be obtained by replacing acetyl chloride with 1-pyrrolidinecarbonyl chloride; and 048 can be obtained by replacing acetyl chloride with ethyl chloroformate.

[0264] Example 11:

[0265]

[0266] Compound 006 (200 mg) was added to a DCM / TFA (5 ml / 0.3 ml) solution and heated under reflux for 2 h. After cooling, an aqueous sodium bicarbonate solution was added to the reaction solution to adjust the pH to about 4. The layers were separated, and the aqueous layer was extracted with ethyl acetate. The organic phases were combined, washed with water, dried over anhydrous magnesium sulfate, concentrated, and purified by preparative HPLC to obtain the target product 034. 1H NMR(400MHz,CD3OD)δ7.95–6.95(m,10H),5.22(d,J=11.1Hz,1H),4.25(dd,J=13.8,10.4Hz,1H), 4.05(p,J=6.9Hz,1H),3.62–3.52(m,1H),3.51–3.40(m,2H),3.00(d,J=13.5Hz,1H),2.63(d,J=1 3.6Hz,1H),2.34(t,J=13.7Hz,1H),2.15(dt,J=14.1,7.0Hz,1H),2.05(dd,J=13.6,2.9Hz,1H),1 .48(t,J=7.1Hz,6H),1.39(s,3H),0.68(d,J=6.6Hz,3H),0.50(d,J=6.9Hz,3H).LC-MS: M+1=610.2

[0267] Example 12:

[0268] Referring to Example 4, compound 054 can be obtained by replacing compound 11 with compound 034. 1 H NMR (400MHz, DMSO-d6) δ12.82(s,1H),9.61(s,1H),8.27(d,J=8.2Hz,1H),7.58(dd,J=8.4,1.8Hz,1H),7.52(d,J=1.8Hz,1H),7.50–7 .06(m,6H),6.96(d,J=7.5Hz,1H),6.84(s,1H),5.13(d,J=11.1Hz,1H),4.08(dd,J=14.0,10.3Hz,1H),3.94(q,J=7.0Hz,1H),3.87(s ,3H),3.57(t,J=12.8Hz,1H),3.47(d,J=13.4Hz,1H),3.22(t,J=9.6Hz,1H),3.11(d,J=13.5Hz,1H),2.84(d,J=13.3Hz,1H),2.18–1. 98 (m, 4H), 1.37 (d, J = 6.7Hz, 3H), 1.33 (d, J = 6.9Hz, 2H), 1.29 (s, 2H), 0.55 (d, J = 6.5Hz, 3H), 0.41 (d, J = 6.9Hz, 3H). LC-MS: M+1 = 759.2.

[0269] Example 13:

[0270] Referring to Example 4, compounds 042, 043, 049, 050, 051, 052 and 053 can also be synthesized.

[0271] Table B

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281] Activity Test Example 1 Homogeneous Time-Resolved Fluorescence Assay (HTRF Assay)

[0282] Standard assay conditions for the in vitro HTRF assay consisted of 1 mM DTT, 0.1% BSA, 2.5 nM GST-hMDM2 (aa1-188), 5 nM biotinylated-p53 (aa1-83), 1.8 nM SA-XLent (Cisbio; Bedford, MA), 0.6 nM anti-GST cryptate monoclonal antibody (Cisbio; Bedford, MA), and 200 mM KF in 1X PBS buffer, pH 7.4, in a total reaction volume of 50 μl in a black 384-well Costar polypropylene plate. Amino acid residues 1-188 of human MDM2 were expressed in E. coli as an amino-terminal glutathione-S-transferase (GST) fusion protein (GST-hMDM2). Residues 1-83 of human p53 were expressed in E. coli as an amino-terminal AviTag. <tm>-TrxA-6xHis fusion protein (biotinylated p53). Each protein was isolated from cell homogenate by affinity chromatography.

[0283] Specifically, 10 μL of GST-hMDM2 was incubated with 10 μL of diluted compound (various concentrations, serial dilutions) in 10% DMSO at room temperature for 20 minutes. 20 μL of biotinylated-p53 was added to the GST-hMDM2 + compound mixture and then incubated at room temperature for 60 minutes. 10 μL of detection buffer consisting of SA-XLent, anti-GST cryptate antibody, and KF was added to the GST-hMDM2, biotinylated-p53, and compound reaction and allowed to equilibrate at room temperature and maintain equilibrium for >4 hours. The final concentration of DMSO in the reaction was 2%. Time-resolved fluorescence readings were measured on a microplate multilabel reader. Percent inhibition was calculated relative to nutlin-3.

[0284] When the potency of the MDM2 inhibitor was increased, a modified HTRF assay (HTRF2 assay) was performed. All assay conditions were the same as above except for the following reagent concentration changes: 0.2 nM GST-hMDM2 (1-188), 0.5 nM biotinylated-p53 (1-83), 0.18 nM SA-XLent, and 100 mM KF.

[0285] The results are listed in the following Table 1, where + represents <10 nm, ++ represents 10 to 100 nm, and +++ represents >100 nm.

[0286] Table 1 HTRF analysis

[0287]

[0288]

[0289] Biological Test Example 2p21 Assay

[0290] Inhibition of the interaction between hMDM2 and p53 leads to activation of the p53 pathway via stabilization and accumulation of p53. p53 activates the transcription of many genes, one of which is p21 <waf1 cip1>To assess the potency of hMDM2 inhibitors, quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to measure p21 transcript levels in compound-associated cells relative to dimethyl sulfoxide (DMSO)-treated control cells.

[0291] On day 1, SJSA-1 cells were plated at 3×10 4 Cells were seeded at a density of 100 cells / well in 100 μl of growth medium (RPMI 1640; 10 mM HEPES; 1 mM sodium pyruvate; 1× penicillin-streptomycin-glutamine (PSQ); and 10% fetal bovine serum (all reagents were obtained from Invitrogen; Carlsbad, CA)) in a 96-well cell culture plate. The cells were cultured overnight at 37°C and 5% CO2.

[0292] On day 2, hMDM2 inhibitors were serially diluted in DMSO (Sigma-Aldrich; St. Louis, MO). 5 μl of each compound dilution was added to 245 μl of filtered assay medium (RPMI1640, 10 mM HEPES, 1 mM sodium pyruvate, and 1X PSQ) containing 10% FBS. Alternatively, the assay was performed in the presence of 10% human serum or 10% mouse serum, or in the absence of any serum. Growth medium was removed from the seeded SJSA-1 cells and replaced with 100 μl / well of assay medium. 100 μl of medium containing the diluted inhibitor was then added to each well to a final volume of 200 μl. Compounds were dose-titrated to yield final concentrations ranging from 0.049 μM to 50 μM, with the addition of a DMSO control. The cells were incubated in the presence of the inhibitor at 37°C and 5% CO₂ for 7 hours. At the end of the incubation period, the medium was removed from the cells, and the plates were stored at −80°C.

[0293] On day 3, total RNA was purified from inhibitor- and DMSO-treated SJSA-1 cells using a Qiagen BioRobot Universal workstation following the RNeasy 96 BioRobot 8000 kit protocol from the manufacturer (Qiagen; Valencia, CA).

[0294] To measure the level of p21 transcripts present, qRT-PCR was used. The levels of p21 and the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), were determined in technical replicates from total RNA from each inhibitor- or DMSO-treated well. qRT-PCR reactions were determined using the relative quantification (ΔΔCt) method on an Applied Biosystems Prism 7900HT instrument using the following cycling conditions: 48°C, 30 minutes, followed by 95°C, 10 minutes, and then 40 cycles consisting of 95°C, 15 seconds, and 60°C, 1 minute. Data were analyzed using Applied Biosystems SDS2.2 software using GAPDH as an endogenous control and DMSO-treated samples as a calibrator. SDS2.2 software calculated the relative quantification (RQ) or increase in p21 levels relative to the DMSO control for each treated sample. The maximum (100%) p21 induction fold was defined by the maximum value of the fitted curve for the reference compound. The p21 fold induction at each inhibitor dose tested was converted to a value representing a percentage of the maximum value. Dose-response curves were generated using XLFit software (ID Business Solutions, Alameda, CA) to calculate the IC of each inhibitor tested. 50 Transit value.

[0295] The results are listed in Table 2 below, where + represents <1 μM, ++ represents 1 to 10 μM, and +++ represents >10 μM.

[0296] Table 2 Cell analysis (SJSA-1 cells)

[0297]

[0298]

[0299] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto. < / tm>

Claims

1. A compound represented by the following formula I or a pharmaceutically acceptable salt thereof: in, The compound of formula I has the structure described in formula III below: Ra and Rb are each independently unsubstituted or halogenated phenyl; Rc is selected from the following group: unsubstituted C1-C6 alkyl; X is C=O; Z1 is selected from the following groups: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C 10 Aryl; the "substituted" refers to substituted by one or more substituents selected from the group consisting of halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, hydroxy, amino, carboxyl, -CN; Q is selected from the following group: 、 、 or ; m and p are each independently 1; Each Z2 or Z3 is independently selected from the group consisting of: substituted or unsubstituted C1-C7 alkylene; Z4 is selected from 、 ; wherein, R1 is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, cyano, -C(=O)-NRdRe, -C(=O)-substituted or unsubstituted C1-C6 alkoxy, -C(=O)-substituted or unsubstituted C1-C6 alkyl, -C(=O)-substituted or unsubstituted C3-C10 cycloalkyl, -C(=O)-substituted or unsubstituted C2-C6 alkenyl, -C(=O)-substituted or unsubstituted C2-C6 alkynyl; Rd and Re are each independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C10 cycloalkyl; or said Rd and Re together with adjacent N atoms form a 4-10 membered heterocyclic ring containing 1-2 nitrogen atoms and 0-2 S or O atoms; R2 is selected from the following groups: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl; Unless otherwise specified, the term "substituted" refers to substitution by one or more substituents selected from the group consisting of halogen, C1-C6 alkyl, and halogenated C1-C6 alkyl; The additional condition is that when Z4 is When Q is .

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The compound of formula I has the structure described in formula IV below: IV Wherein, the definitions of each group are as described in claim 1.

3. A method for preparing the compound of formula I according to claim 1, characterized in that: The method comprises or is carried out by the following steps (1), (2) or (3): Step (1): Step (2): Step (3): Wherein, the definition of each group is as described in claim 1.

4. A pharmaceutical composition, characterized in that The invention comprises (1) the compound according to claim 1 or a pharmaceutically acceptable salt thereof; and (2) a pharmaceutically acceptable carrier.

5. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 4, characterized in that: Used for preparing a pharmaceutical composition for preventing and / or treating diseases related to the activity or expression of MDM2.

6. The use according to claim 5, characterized in that The pharmaceutical composition is used to treat a disease selected from the group consisting of bladder cancer, breast cancer, colon cancer, rectal cancer, kidney cancer, liver cancer, lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, prostate cancer and skin cancer, lymphoid lineage hematopoietic system tumors, myeloid lineage hematopoietic system tumors, tumors of mesenchymal origin, tumors of the central and peripheral nervous systems, melanoma, seminoma, teratoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, follicular thyroid carcinoma, Kaposi's sarcoma, Endometrial cancer, head and neck cancer, glioblastoma, malignant ascites, hematopoietic cancer, thyroid hyperplasia, cysts, asthma, chronic obstructive pulmonary disease, emphysema, psoriasis, contact dermatitis, conjunctivitis, allergic rhinitis, systemic lupus erythematosus, ulcerative colitis, Crohn's disease, multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, Alzheimer's disease, atherosclerosis, Huntington's disease, inflammatory diseases, hypoxia, ulcers, viral infections, bacterial infections, and bacterial sepsis.

7. An MDM2 inhibitor, characterized in that The inhibitor comprises the compound according to claim 1, or a pharmaceutically acceptable salt thereof.

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