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WO2023241738A2 - 1,4-benzodiazepine compound and use thereof in preparation of anti-tumor drug - Google Patents

1,4-benzodiazepine compound and use thereof in preparation of anti-tumor drug Download PDF

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Publication number
WO2023241738A2
WO2023241738A2 PCT/CN2023/112421 CN2023112421W WO2023241738A2 WO 2023241738 A2 WO2023241738 A2 WO 2023241738A2 CN 2023112421 W CN2023112421 W CN 2023112421W WO 2023241738 A2 WO2023241738 A2 WO 2023241738A2
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preparation
cancer
compound
dmso
nmr
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PCT/CN2023/112421
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French (fr)
Chinese (zh)
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WO2023241738A3 (en
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孙逊
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复旦大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/551—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having two nitrogen atoms, e.g. dilazep
    • A61K31/5513—1,4-Benzodiazepines, e.g. diazepam or clozapine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
    • C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
    • C07D243/14—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines
    • C07D243/16—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals
    • C07D243/18—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals substituted in position 2 by nitrogen, oxygen or sulfur atoms
    • C07D243/24—Oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
    • C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
    • C07D243/14—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines
    • C07D243/16—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals
    • C07D243/18—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals substituted in position 2 by nitrogen, oxygen or sulfur atoms
    • C07D243/24—Oxygen atoms
    • C07D243/28—Preparation including building-up the benzodiazepine skeleton from compounds containing no hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C07D491/113—Spiro-condensed systems with two or more oxygen atoms as ring hetero atoms in the oxygen-containing ring

Definitions

  • the invention belongs to the fields of medicinal chemistry and medical technology and pharmaceuticals, and relates to 1,4-benzodiazepine.
  • Compounds and their use in preparing anti-tumor drugs, specifically involving 1,4-benzodiazepine The use of analog compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, or pharmaceutical compositions thereof and medically acceptable carriers in the preparation of ANXA3 degradation agents and in the preparation of prevention and/or treatment of cancer uses in medicines.
  • Cancer is a common malignant tumor in clinical practice, characterized by rapid disease progression and high mortality.
  • the latest report from the World Health Organization shows that there will be 19.3 million new cancer patients and 10 million deaths from cancer in the world in 2020 (CA: a cancer journal for clinicians, 2021, 71(3):209-249). It is expected that by 2040, the number of new cancer patients worldwide will reach 28.4 million, an increase of 47% from 2020. Therefore, cancer has become one of the major diseases that seriously endangers human health around the world.
  • Targeted drugs have become a hot area of anti-tumor drug research and development in recent years because they can specifically act on abnormally expressed proteins in tumor cells and thereby induce specific death of tumor cells without affecting surrounding normal cells.
  • targeted drugs can be mainly divided into various types such as macromolecular antibodies, small molecule inhibitors, small molecule agonists, and small molecule degraders. So far, small molecule inhibitors are one of the important types of targeted therapy, but they still have obvious shortcomings and limitations.
  • small molecule degraders have subverted the previous concept of "difficult to drug" targets. These small molecule degraders not only have small dosages and high selectivity, but also greatly reduce off-target effects (Cell Chemical Biology. 2017, 24(9): 1181-1190). In addition, small molecule degraders have unique catalytic mechanisms, especially the ability to target traditional "difficult-to-drug" targets and solve pain points such as drug resistance.
  • small molecule degraders have become a hot spot and frontier field in the current research and development of new drugs, and has achieved rapid development. Judging from the clinical research and development pipeline that has been advanced, small molecule degraders have excellent performance in terms of efficacy and safety, and have broad application value especially in the field of malignant tumor treatment. Therefore, the development of small molecule degraders with new targets has become an important research direction in the research and development of anti-tumor drugs, which is expected to meet the urgent unmet clinical needs of cancer patients.
  • Annexin A3 is a member of the Annexin (ANX) family. It can bind to acidic phospholipids in a Ca 2+ -dependent manner and participate in a series of Ca 2+ changes on the cell membrane surface. Dependent physiological activities, including vesicle trafficking, membrane fusion during exocytosis, signal transduction, formation of Ca 2+ channels, and interactions between cytoskeletal proteins (Nature reviews Molecular cell biology, 2005, 6(6) :449-461.), but the biochemical function of the ANXA3 protein is largely unknown. Normally, ANXA3 protein is expressed in differentiated cells of myeloid cell lines.
  • ANXA3 protein in many cancers, such as breast cancer, There is scientific evidence of abnormal expression of ANXA3 protein in ovarian cancer, lung adenocarcinoma, prostate cancer, kidney cancer, colon cancer, pancreatic cancer and liver cancer, and the abnormal expression of ANXA3 protein is closely related to the progression of these cancers (Clinical and Translational Oncology,2013,15(2):106-110). For example, many studies have been dedicated to revealing the important role of ANXA3 in the development and progression of breast cancer.
  • ANXA3 is highly expressed in breast tumor samples, especially in many triple-negative breast cancer (TNBC) tumor samples, with a positive expression rate of 79.66%, which is significantly higher than that of other types of breast cancer patients, and high expression levels of ANXA3 are closely related to poor prognosis.
  • TNBC triple-negative breast cancer
  • ANXA3 is highly expressed in breast tumor samples, especially in many triple-negative breast cancer (TNBC) tumor samples, with a positive expression rate of 79.66%, which is significantly higher than that of other types of breast cancer patients, and high expression levels of ANXA3 are closely related to poor prognosis.
  • TNBC triple-negative breast cancer
  • ANXA3 silencing the expression of ANXA3 not only significantly inhibited the growth of TNBC transplanted tumors at the animal level in vivo, but also effectively inhibited the lung metastasis of TNBC orthotopic transplanted tumors, while reducing the resistance of TNBC cells to the chemotherapy drug doxorubicin.
  • Medicinal properties Pathology-Research and Practice, 2018, 214 (10): 1719-1725; Cell Death & Disease, 2018, 9 (2): 1-11). It is speculated that the development of selective degraders targeting ANXA3 to induce ANXA3 protein degradation may be a promising new anti-tumor treatment strategy.
  • ANXA3 as a new class of drug targets, there have been no reports so far of small chemical molecules that can target and degrade ANXA3 and exhibit anti-tumor pharmacological activity.
  • this application provides 1,4-benzodiazepine Compounds and their use in preparing anti-tumor drugs, specifically involving 1,4-benzodiazepine
  • This application discovered the original 1,4-benzodiazepine through structure-activity relationship screening of the self-constructed small molecule compound library targeting ANXA3 binding activity and degradation activity, as well as research on anti-tumor cell activity.
  • the compound can target and degrade ANXA3 and exhibit anti-tumor pharmacological activity.
  • the first object of the present invention is to provide 1,4-benzodiazepine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof;
  • the 1,4-benzodiazepine Compounds are compounds or salts with a structure shown in formula (I),
  • R 1 is selected from a hydrogen atom or a methyl group
  • X is a linking group, selected from: Where, R 2 is selected from:
  • R 3 to R 7 are each independently selected from hydrogen, halogen, methoxy, trifluoromethoxy, trifluoromethyl, methyl, ethyl, dimethylamino, nitro, cyano or acetyl;
  • R 8 to R 11 are each independently selected from hydrogen, halogen, cyano, nitro, trifluoromethyl or methoxy;
  • R' 8 -R' 11 are each independently selected from hydrogen, halogen, cyano, nitro, trifluoromethyl or methoxy;
  • R 12 to R 14 are each independently selected from hydrogen or halogen
  • R' 12 to R' 14 are each independently selected from hydrogen or trifluoromethyl
  • R 12 to R” 14 are each independently selected from hydrogen
  • Aromatic five-membered heterocycle its structure is:
  • A is selected from nitrogen, oxygen or sulfur
  • B is selected from carbon or nitrogen
  • C is selected from carbon or nitrogen
  • D is selected from carbon or nitrogen
  • E is selected from nitrogen or oxygen
  • F is selected from carbon or nitrogen
  • R 15 to R 18 are each independently selected from hydrogen, methyl or trifluoromethyl
  • R' 15 to R' 17 are each independently selected from hydrogen or methyl
  • G is selected from carbon or nitrogen
  • R 19 is selected from methoxy or ethylene glycol.
  • pharmaceutically acceptable salts refer to salts of compounds that are suitable for use within the range of reliable medical evaluation. Contact with tissues of humans or lower animals without undue toxicity, irritation and allergic reactions, etc., with a reasonable ratio of benefits to risks, usually water or oil soluble or dispersible, and can be effectively used for its intended use.
  • Some of the compounds of the present invention or their stereoisomers contain basic groups such as amine groups, which can form salts with acids, and can form acid salts with inorganic and/or organic acids, including zwitterionic salts (inner salts), and Quaternary ammonium salts, such as alkylammonium salts.
  • These salts may be obtained directly from the final isolation and purification of the compound, or its stereoisomers. It can also be obtained by appropriately mixing the compound, or its stereoisomer, with a certain amount of acid (for example, equivalent amounts).
  • These salts may form a precipitate in the solution and be collected by filtration, or may be recovered after evaporation of the solvent, or may be obtained by reacting in an aqueous medium and then freeze-drying.
  • Pharmaceutically acceptable salts described in the present invention include organic acid salts such as citrate, benzenesulfonate, acetate, propionate, succinate, oxalate, malate, and succinate. acid salt, fumarate, maleate, tartrate or trifluoroacetate, etc.; inorganic acid salts such as hydrochloride, sulfate, hydrobromide, hydrofluoride, hydroiodide, hydrogen iodide, etc. Chlorate, phosphate, etc.; or with amino acids, such as glutamic acid or aspartic acid, can form glutamate or aspartate.
  • organic acid salts such as citrate, benzenesulfonate, acetate, propionate, succinate, oxalate, malate, and succinate.
  • acid salt fumarate, maleate, tartrate or trifluoroacetate, etc.
  • inorganic acid salts such as hydrochloride, sulfate, hydrobromide, hydro
  • 1,4-benzodiazepine of the present invention Solvates of similar compounds also fall within the protection scope of the present invention.
  • the solvents are preferably water, alcohol or alcohol-water mixtures.
  • the alcohol here is methanol or ethanol.
  • the second object of the present invention is to provide 1,4-benzodiazepine represented by formula (I)
  • the binding activity test based on surface plasmon resonance showed that the 1,4-benzodiazepine represented by formula (I)
  • the compound has submicromolar ANXA3 binding activity, and the activity results are shown in Table 1.
  • Experiments based on Western blot testing showed that the compound can induce ANXA3 protein degradation in TNBC cells, with activity at the micromolar level, and the results are shown in Table 1.
  • the degradation activity results of representative isomer compounds I-9a19, (R)-I-9a19 or (S)-I-9a19 are shown in Figure 1.
  • a further object of the present invention is to provide 1,4-benzodiazepine represented by formula (I)
  • Table 1 In vitro anti-tumor activity tests on various tumor cells show that the compound has anti-tumor activity at the micromolar level, and the activity results are shown in Table 1.
  • compound I-9a19 can significantly inhibit the cloning, migration and invasion of TNBC cells.
  • the activity results are shown in Figure 2.
  • compound I-9a19 showed effective therapeutic effects and induced the degradation of ANXA3 protein in tumor tissue in the in vivo TNBC transplanted tumor model.
  • the activity results are shown in Figure 3.
  • the above-mentioned medicines may also contain one or more pharmaceutically acceptable carriers.
  • the carriers include conventional diluents, excipients, fillers, binders, wetting agents, disintegrating agents, and absorbers in the pharmaceutical field. Accelerators, surfactants, adsorption carriers, lubricants, etc., and flavors, sweeteners, etc. can also be added if necessary.
  • the present invention also provides an anti-tumor pharmaceutical composition that exerts an anti-tumor effect by acting as an annexin ANXA3 degrading agent.
  • the anti-tumor pharmaceutical composition is a tablet, capsule, pill, injection, sustained-release preparation, or spray. Or nano drug delivery system.
  • the beneficial effect of the present invention lies in the 1,4-benzodiazepine provided Compounds are a class of structurally novel ANXA3
  • the small molecule degrader not only has sub-micromolar ANXA3 binding effect, but also shows obvious ANXA3 degradation activity in TNBC cells and animal levels. Moreover, it can significantly inhibit the proliferation, cloning, migration and invasion of TNBC cells in vitro.
  • TNBC cells through the orthotopic xenograft tumor model of human MDA-MB-231 cells in female BALB/c nude mice, it has been confirmed that the anti-tumor function is better in vivo.
  • TNBC has good potential and application prospects, and can be further prepared into drugs for the treatment of cancers, such as breast cancer, cervical cancer, ovarian cancer, intestinal cancer, gastric cancer, pancreatic cancer, lung cancer, esophageal cancer, and liver cancer.
  • cancers such as breast cancer, cervical cancer, ovarian cancer, intestinal cancer, gastric cancer, pancreatic cancer, lung cancer, esophageal cancer, and liver cancer.
  • leukemia and melanoma preferably, leukemia and melanoma.
  • the anti-tumor drug is an anti-triple-negative breast cancer drug.
  • 1,4-benzodiazepine provided by the invention
  • the structures of similar compounds and their pharmaceutically acceptable salts have not been reported by Scifinder. They are a class of ANXA3 small molecule degraders with novel structures. This type of compound exerts anti-tumor therapeutic effects in vitro and in vivo by directly binding to ANXA3 and inducing its degradation. It solves the lack of small molecule drugs that can directly bind to ANXA3 and exhibit anti-tumor activity in the existing technology, and provides a basis for clinical use. It provides new treatment options and medication regimens for the treatment of tumors.
  • Figure 1 Detection of the effects of compounds I-9a19, (R)-I-9a19 and (S)-I-9a19 on ANXA3 protein levels;
  • Figure 3 In vivo anti-TNBC efficacy study of compound I-9a19 and detection of ANXA3 degradation in tumor tissue.
  • Compound I-9a3 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8a3.
  • 1 H NMR 400MHz, DMSO-d 6 ) ⁇ 11.53(s,1H),11.20(s,1H),10.73(s,1H),8.09(s,1H),7.68(s,5H),7.59- 7.43(m,5H),7.30(s,2H),5.94(s,1H),4.25(s,2H),3.37(s,3H) .
  • Compound I-9b1 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8b1.
  • 1 H NMR 400MHz, DMSO-d 6 ) ⁇ 11.54(s,1H),11.14(s,1H),10.69(s,1H),8.28(s,2H),7.92(s,2H),7.79( s,2H),7.68(s,3H),7.51(m,8H),7.30(s,3H),7.11(s,1H),5.94(s,1H),4.24(s,3H),3.37(s ,4H) .
  • Compound I-9d1 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8d1.
  • Compound I-10a5 was synthesized according to the method of step 1.3 in Example 1.
  • 1 H NMR 400MHz, DMSO-d 6 ) ⁇ 11.49(s,1H),11.16(s,1H),11.04-10.96(m,1H),10.71(s,1H),8.31(s,1H), 7.96(s,1H),7.82(s,1H),7.66(s,1H),7.58-7.45(m,5H),7.36-7.22(m,3H),7.14(s,1H),5.92(s, 1H),4.30(s,2H) .
  • 1,4-Benzodiazepine according to the present invention Binding activity test of similar compounds to ANXA3 protein.
  • 1,4-Benzodiazepine according to the present invention Test of the degradation activity of similar compounds on ANXA3 protein.
  • protease inhibitor (Swiss Roche Company), BCA protein quantitative detection kit (American Thermo Fisher Scientific Company), ECL chromogenic solution (WBKLS0500, American Milipore Company), Tubulin antibody (HC101-02, Beijing Full Gold Biological Company), ANXA3 antibody (11804-1-AP, Proteinteach Company of the United States), secondary antibodies (HS201-01, HS101-01, Beijing Quanshijin Biotechnology Company).
  • Experimental steps 1.5*10 4 cells were seeded in a 6cm culture dish. After 24 hours, compound formula I solution was added to the cells to form the experimental group. Culture medium containing 0.1% DMSO was added to the cells as a control group. After incubation for 24 hours, cells were collected, RIAP solution was added and lysed on ice for 30 minutes, and then centrifuged at 12,000 rpm for 15 minutes at 4°C and the supernatant was collected. The BCA kit detects the total protein concentration. The total protein loading amount is 50 ⁇ g. The stacking gel electrophoresis voltage is 80V and the separation gel electrophoresis voltage is 120V. Then the film is transferred at a constant voltage of 110V for 90 minutes.
  • 1,4-Benzodiazepine according to the present invention In vitro experiments on the inhibition of tumor cell viability by similar compounds.
  • MTT American Sigma Company
  • fetal calf serum Ausbian Company, Australia
  • DMEM culture medium American Gibco Company
  • Compound I-9a19 of the present invention inhibits tumor cell colony formation in vitro.
  • the main reagents in the experiment Matrigel Matrigel (BD Company, USA), RIPA lysis buffer (Thermo Fisher Scientific Company, USA).
  • mice Inject 1 ⁇ 10 6 cells into the fourth pair of breast pads of female BALB/c nude mice. After the tumor volume reaches approximately 100 mm 3 , the nude mice are randomly divided into blank solvent group and control drug docetaxel (DTX). ) group and compound I-9a19 treated group. Administration was started on the first day, and after 19 consecutive days of administration, the nude mice were sacrificed, tumor tissues were obtained, weighed, and TGI was calculated. Then take 10 mg of tumor, add RIPA lysis buffer, and then centrifuge, collect the supernatant, use the BCA method for protein quantification, and perform subsequent operations according to the Western blot experiment.
  • DTX control drug docetaxel
  • the binding activity test based on surface plasmon resonance shows that the 1,4-benzenediazepine represented by formula (I)
  • the compound has submicromolar ANXA3 binding activity, and the activity results are shown in Table 1.
  • Experiments based on Western blot testing showed that the compound can induce ANXA3 protein degradation in TNBC cells, with activity at the micromolar level, and the results are shown in Table 1.
  • the degradation activity results of representative isomer compounds I-9a19, (R)-I-9a19 or (S)-I-9a19 are shown in Figure 1.
  • Affinity K D , degradation activity DC 50 and half inhibitory concentration IC 50 *: >100 ⁇ M, **: 10-100 ⁇ M, ***: 1-10 ⁇ M, ****: ⁇ 1 ⁇ M.

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Abstract

The invention belongs to the technical field of pharmaceutical chemistry and medicines, relating to a 1,4-benzodiazepine compound and a use thereof in the preparation of an antitumor drug, in particular, to a use of the 1,4-benzodiazepine compound in the preparation of an annexin A3 (ANXA3) degradation agent and a use of the 1,4-benzodiazepine compound in the preparation of a drug for treating cancers. In particular, disclosed is a use of the 1,4-benzodiazepine compound, or a pharmaceutically acceptable salt or stereoisomer thereof, or a pharmaceutical composition using the compound as an active ingredient in the preparation of a drug for preventing and treating tumors. The compound can bind to ANXA3 protein, induce ANXA3 protein degradation, inhibit the proliferation, cloning, migration, invasion and other functions of tumor cells, and exert an anti-tumor treatment effect in vivo. The method can be used for treating solid tumors, blood tumors and other diseases, and the involved tumors comprise breast cancer, cervical cancer, ovarian cancer, intestinal cancer, gastric cancer, pancreatic cancer, lung cancer, esophageal cancer, liver cancer, leukemia and melanoma.

Description

1,4-苯二氮*类化合物及其在制备抗肿瘤药物中的用途1,4-Benzodiazepine compounds and their use in the preparation of anti-tumor drugs 技术领域Technical field
本发明属于药物化学和医药技术制药领域，涉及1,4-苯二氮类化合物及其在制备抗肿瘤药物中的用途，具体涉及1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体、或其与医学上可接受的载体组成的药物组合物在制备ANXA3降解剂中的用途以及在制备预防和/或治疗癌症的药物中的用途。The invention belongs to the fields of medicinal chemistry and medical technology and pharmaceuticals, and relates to 1,4-benzodiazepine. Compounds and their use in preparing anti-tumor drugs, specifically involving 1,4-benzodiazepine The use of analog compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, or pharmaceutical compositions thereof and medically acceptable carriers in the preparation of ANXA3 degradation agents and in the preparation of prevention and/or treatment of cancer uses in medicines.
背景技术Background technique
癌症是临床上常见的一种恶性肿瘤病变，具有病情进展快、死亡率高等特点。世界卫生组织最新报告显示，2020年全球有1930万癌症新增患者，1000万死于癌症(CA:a cancer journal for clinicians,2021,71(3):209-249)。预计到2040年，全球癌症新增患者将达到2840万，比2020年增加47％。因此，癌症已成为严重危害全球人类身体健康的重大疾病之一。Cancer is a common malignant tumor in clinical practice, characterized by rapid disease progression and high mortality. The latest report from the World Health Organization shows that there will be 19.3 million new cancer patients and 10 million deaths from cancer in the world in 2020 (CA: a cancer journal for clinicians, 2021, 71(3):209-249). It is expected that by 2040, the number of new cancer patients worldwide will reach 28.4 million, an increase of 47% from 2020. Therefore, cancer has become one of the major diseases that seriously endangers human health around the world.
目前肿瘤的治疗方式有外科治疗、放射治疗和药物治疗等三种手段。其中，药物治疗经历了由化学治疗到靶向治疗，再到免疫治疗的突破性进展，使得肿瘤患者的生存状况得到了显著的改善。靶向药物由于能够特异性作用于肿瘤细胞的异常表达蛋白，进而诱导肿瘤细胞特异性死亡，而不会波及周围正常细胞，已成为近年来抗肿瘤药物研发的热点领域。根据药物的性质及其作用机制的不同，可将靶向药物主要分为大分子抗体、小分子抑制剂、小分子激动剂和小分子降解剂等多种类型。迄今为止，小分子抑制剂是靶向治疗的重要类型之一，但仍存在明显的不足与局限性，如无法完全阻断蛋白的生物学功能，不能高效作用于蛋白靶点的活性结合口袋，以及缺乏选择性易引起脱靶效应和耐药性等。近年来，小分子降解剂颠覆了先前认为“难以成药”靶点的观念。这些小分子降解剂不仅用量小，选择性高，而且大大降低了脱靶效应(Cell Chemical Biology.2017,24(9):1181-1190)。此外，小分子降解剂具有独特的催化机制，尤其是能够靶向传统“难以成药”靶点并解决药物耐药性等痛点。因而，小分子降解剂的研究已成为目前新药研发的热点和前沿领域，并已取得快速的发展。从已在推进的临床研发管线来看，小分子降解剂在药效上和安全性方面都表现优异，尤其在恶性肿瘤治疗领域拥有广阔的应用价值。因此，开发新靶点的小分子降解剂已成为抗肿瘤药物研发的重要研究方向，有望满足癌症患者临床未被满足的迫切需求。Currently, there are three types of cancer treatments: surgical treatment, radiotherapy and drug treatment. Among them, drug treatment has experienced breakthrough progress from chemotherapy to targeted therapy to immunotherapy, which has significantly improved the living conditions of tumor patients. Targeted drugs have become a hot area of anti-tumor drug research and development in recent years because they can specifically act on abnormally expressed proteins in tumor cells and thereby induce specific death of tumor cells without affecting surrounding normal cells. According to the nature of the drug and its mechanism of action, targeted drugs can be mainly divided into various types such as macromolecular antibodies, small molecule inhibitors, small molecule agonists, and small molecule degraders. So far, small molecule inhibitors are one of the important types of targeted therapy, but they still have obvious shortcomings and limitations. For example, they cannot completely block the biological function of the protein and cannot efficiently act on the active binding pocket of the protein target. And the lack of selectivity can easily lead to off-target effects and drug resistance. In recent years, small molecule degraders have subverted the previous concept of "difficult to drug" targets. These small molecule degraders not only have small dosages and high selectivity, but also greatly reduce off-target effects (Cell Chemical Biology. 2017, 24(9): 1181-1190). In addition, small molecule degraders have unique catalytic mechanisms, especially the ability to target traditional "difficult-to-drug" targets and solve pain points such as drug resistance. Therefore, the research on small molecule degraders has become a hot spot and frontier field in the current research and development of new drugs, and has achieved rapid development. Judging from the clinical research and development pipeline that has been advanced, small molecule degraders have excellent performance in terms of efficacy and safety, and have broad application value especially in the field of malignant tumor treatment. Therefore, the development of small molecule degraders with new targets has become an important research direction in the research and development of anti-tumor drugs, which is expected to meet the urgent unmet clinical needs of cancer patients.
据文献研究显示，膜联蛋白A3(Annexin A3，ANXA3)是膜联蛋白(Annexin,ANX)家族成员之一，能够以Ca2+依赖的方式与酸性磷脂结合，参与细胞膜表面一系列Ca2+依赖的生理活动，包括囊泡运输、胞吐过程中的膜融合、信号转导、Ca2+通道的形成以及细胞骨架蛋白之间的相互作用(Nature reviews Molecular cell biology,2005,6(6):449-461.)，但ANXA3蛋白的生化功能在很大程度上是未知的。正常情况下，ANXA3蛋白表达于髓样细胞系的分化细胞中。但近年来临床和生物医学基础研究发现，在多种癌症如乳腺癌、 卵巢癌、肺腺癌、前列腺癌、肾癌、结肠癌、胰腺癌和肝癌等中都有ANXA3蛋白异常表达的科学证据，且ANXA3蛋白的异常表达与这些癌症的病情进展密切相关(Clinical and Translational Oncology,2013,15(2):106-110)。例如，已有多项研究致力于揭示ANXA3在乳腺癌的发生发展过程中的重要作用。ANXA3在乳腺肿瘤样本，尤其是在很多三阴性乳腺癌(TNBC)肿瘤样本中呈现高表达，阳性表达率为79.66％，显著高于其他类型乳腺癌患者，并且ANXA3的高表达水平与不良预后密切相关(BioMed Research International,2017,2017:1-7)。基于RNA干扰技术沉默ANXA3的表达在细胞水平上显著抑制了TNBC的增殖、迁移和侵袭活性，并诱导细胞凋亡(Oncology reports,2017,37(1):388-398；Clinical breast cancer,2018,18(4):713-719)。重要的是，沉默ANXA3的表达不仅能够在体内动物水平上显著抑制TNBC移植瘤的生长，还有效抑制了TNBC原位移植瘤的肺转移，同时降低了TNBC细胞对化疗药多柔比星的耐药性(Pathology-Research and Practice,2018,214(10):1719-1725；Cell Death&Disease,2018,9(2):1-11)。由此推测，研发靶向ANXA3的选择性降解剂用来诱导ANXA3蛋白降解，可能是一种有前景的抗肿瘤治疗新策略。然而，关于ANXA3作为一类全新的药物靶点，迄今未见报道能够靶向降解ANXA3并表现出抗肿瘤药理活性的化学小分子。According to literature research, Annexin A3 (ANXA3) is a member of the Annexin (ANX) family. It can bind to acidic phospholipids in a Ca 2+ -dependent manner and participate in a series of Ca 2+ changes on the cell membrane surface. Dependent physiological activities, including vesicle trafficking, membrane fusion during exocytosis, signal transduction, formation of Ca 2+ channels, and interactions between cytoskeletal proteins (Nature reviews Molecular cell biology, 2005, 6(6) :449-461.), but the biochemical function of the ANXA3 protein is largely unknown. Normally, ANXA3 protein is expressed in differentiated cells of myeloid cell lines. However, in recent years, clinical and biomedical basic research has found that in many cancers, such as breast cancer, There is scientific evidence of abnormal expression of ANXA3 protein in ovarian cancer, lung adenocarcinoma, prostate cancer, kidney cancer, colon cancer, pancreatic cancer and liver cancer, and the abnormal expression of ANXA3 protein is closely related to the progression of these cancers (Clinical and Translational Oncology,2013,15(2):106-110). For example, many studies have been dedicated to revealing the important role of ANXA3 in the development and progression of breast cancer. ANXA3 is highly expressed in breast tumor samples, especially in many triple-negative breast cancer (TNBC) tumor samples, with a positive expression rate of 79.66%, which is significantly higher than that of other types of breast cancer patients, and high expression levels of ANXA3 are closely related to poor prognosis. Related (BioMed Research International, 2017, 2017: 1-7). Silencing the expression of ANXA3 based on RNA interference technology significantly inhibited the proliferation, migration and invasion activities of TNBC at the cellular level, and induced cell apoptosis (Oncology reports, 2017, 37(1):388-398; Clinical breast cancer, 2018, 18(4):713-719). Importantly, silencing the expression of ANXA3 not only significantly inhibited the growth of TNBC transplanted tumors at the animal level in vivo, but also effectively inhibited the lung metastasis of TNBC orthotopic transplanted tumors, while reducing the resistance of TNBC cells to the chemotherapy drug doxorubicin. Medicinal properties (Pathology-Research and Practice, 2018, 214 (10): 1719-1725; Cell Death & Disease, 2018, 9 (2): 1-11). It is speculated that the development of selective degraders targeting ANXA3 to induce ANXA3 protein degradation may be a promising new anti-tumor treatment strategy. However, regarding ANXA3 as a new class of drug targets, there have been no reports so far of small chemical molecules that can target and degrade ANXA3 and exhibit anti-tumor pharmacological activity.
发明内容Contents of the invention
基于现有技术的不足，本申请提供1,4-苯二氮类化合物及其在制备抗肿瘤药物中的用途，具体涉及1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体、或其与医学上可接受的载体组成的药物组合物在制备ANXA3降解剂中的用途以及在制备预防和/或治疗癌症的药物中的用途。Based on the shortcomings of the existing technology, this application provides 1,4-benzodiazepine Compounds and their use in preparing anti-tumor drugs, specifically involving 1,4-benzodiazepine The use of similar compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, or pharmaceutical compositions thereof and medically acceptable carriers in the preparation of ANXA3 degradation agents and in the preparation of prevention and/or treatment of cancer uses in medicines.
本申请通过对自建的小分子化合物库进行靶向ANXA3结合活性和降解活性的构效关系筛选，以及抗多种肿瘤细胞活性的研究，发现原创的1，4-苯二氮类化合物能够靶向和降解ANXA3并表现出抗肿瘤药理活性的功能。This application discovered the original 1,4-benzodiazepine through structure-activity relationship screening of the self-constructed small molecule compound library targeting ANXA3 binding activity and degradation activity, as well as research on anti-tumor cell activity. The compound can target and degrade ANXA3 and exhibit anti-tumor pharmacological activity.
本发明的第一个目的是提供1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体；所述1，4-苯二氮类化合物是具有如式(I)所示结构的化合物或盐，
The first object of the present invention is to provide 1,4-benzodiazepine compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof; the 1,4-benzodiazepine Compounds are compounds or salts with a structure shown in formula (I),
式(I)中，In formula (I),
R1选自氢原子或甲基；R 1 is selected from a hydrogen atom or a methyl group;
X是链接基团，选自： 其中，R2选自：X is a linking group, selected from: Where, R 2 is selected from:
1)取代苯，其结构为：1) Substituted benzene, its structure is:
其中， in,
R3-R7分别独立地选自氢、卤素、甲氧基、三氟甲氧基、三氟甲基、甲基、乙基、二甲氨基、硝基、氰基或乙酰基；R 3 to R 7 are each independently selected from hydrogen, halogen, methoxy, trifluoromethoxy, trifluoromethyl, methyl, ethyl, dimethylamino, nitro, cyano or acetyl;
2)芳香六元杂环，其结构为：2) Aromatic six-membered heterocycle, its structure is:
其中， in,
R8-R11分别独立地选自氢、卤素、氰基、硝基、三氟甲基或甲氧基；R 8 to R 11 are each independently selected from hydrogen, halogen, cyano, nitro, trifluoromethyl or methoxy;
R’8-R’11分别独立地选自氢、卤素、氰基、硝基、三氟甲基或甲氧基；R' 8 -R' 11 are each independently selected from hydrogen, halogen, cyano, nitro, trifluoromethyl or methoxy;
R12-R14分别独立地选自氢或卤素；R 12 to R 14 are each independently selected from hydrogen or halogen;
R’12-R’14分别独立地选自氢或三氟甲基；R' 12 to R' 14 are each independently selected from hydrogen or trifluoromethyl;
R”12-R”14分别独立地选自氢；R” 12 to R” 14 are each independently selected from hydrogen;
3)芳香五元杂环，其结构为：3) Aromatic five-membered heterocycle, its structure is:
其中， in,
A选自氮、氧或硫；A is selected from nitrogen, oxygen or sulfur;
B选自碳或氮；B is selected from carbon or nitrogen;
C选自碳或氮；C is selected from carbon or nitrogen;
D选自碳或氮；D is selected from carbon or nitrogen;
E选自氮或氧；E is selected from nitrogen or oxygen;
F选自碳或氮；F is selected from carbon or nitrogen;
R15-R18分别独立地选自氢、甲基或三氟甲基；R 15 to R 18 are each independently selected from hydrogen, methyl or trifluoromethyl;
R’15-R’17分别独立地选自氢或甲基；R' 15 to R' 17 are each independently selected from hydrogen or methyl;
4)脂肪杂环或金刚烷，其结构为：4) Aliphatic heterocycle or adamantane, its structure is:
其中， in,
G选自碳或氮；G is selected from carbon or nitrogen;
R19选自甲氧基或缩乙二醇。R 19 is selected from methoxy or ethylene glycol.
本发明中，进一步给出一些式(I)所示的1，4-苯二氮类化合物的具体结构：


In the present invention, some 1,4-benzodiazepines represented by formula (I) are further provided The specific structure of the compound:


本发明中，在药学上可接受的盐类是指在可靠的医药评价范围内，化合物的盐类适于 与人或较低等动物的组织相接触而无不适当的毒性、刺激及过敏反应等，具有相当合理的收益与风险比例，通常是水或油可溶的或可分散的，并可有效地用于其预期的用途。In the present invention, pharmaceutically acceptable salts refer to salts of compounds that are suitable for use within the range of reliable medical evaluation. Contact with tissues of humans or lower animals without undue toxicity, irritation and allergic reactions, etc., with a reasonable ratio of benefits to risks, usually water or oil soluble or dispersible, and can be effectively used for its intended use.
本发明部分化合物或其立体异构体含有碱性基团如胺基等能与酸成盐，可与无机和/或有机酸形成酸式盐，也包括两性离子盐(内盐)，还包括季铵盐，例如烷基铵盐。这些盐可以是在化合物，或其立体异构体，的最后分离和纯化中直接得到。也可以是通过将化合物，或其立体异构体，与一定数量的酸适当(例如等当量)进行混合而得到。这些盐可能在溶液中形成沉淀而以过滤方法收集，或在溶剂蒸发后回收而得到，或在水介质中反应后冷冻干燥制得。Some of the compounds of the present invention or their stereoisomers contain basic groups such as amine groups, which can form salts with acids, and can form acid salts with inorganic and/or organic acids, including zwitterionic salts (inner salts), and Quaternary ammonium salts, such as alkylammonium salts. These salts may be obtained directly from the final isolation and purification of the compound, or its stereoisomers. It can also be obtained by appropriately mixing the compound, or its stereoisomer, with a certain amount of acid (for example, equivalent amounts). These salts may form a precipitate in the solution and be collected by filtration, or may be recovered after evaporation of the solvent, or may be obtained by reacting in an aqueous medium and then freeze-drying.
本发明中所述药效上可接受的盐，包括有机酸盐如枸橼酸盐、苯磺酸盐、乙酸盐、丙酸盐、丁二酸盐、草酸盐、苹果酸盐、琥珀酸盐、富马酸盐、马来酸盐、酒石酸盐或三氟乙酸盐等；无机酸盐如盐酸盐、硫酸盐、氢溴酸盐、氢氟酸盐、氢碘酸盐、氢氯酸盐、磷酸盐等；或与氨基酸，如谷氨酸或天冬氨酸可形成谷氨酸盐或天冬氨酸盐。Pharmaceutically acceptable salts described in the present invention include organic acid salts such as citrate, benzenesulfonate, acetate, propionate, succinate, oxalate, malate, and succinate. acid salt, fumarate, maleate, tartrate or trifluoroacetate, etc.; inorganic acid salts such as hydrochloride, sulfate, hydrobromide, hydrofluoride, hydroiodide, hydrogen iodide, etc. Chlorate, phosphate, etc.; or with amino acids, such as glutamic acid or aspartic acid, can form glutamate or aspartate.
本发明1，4-苯二氮类化合物的溶剂合物也属本发明的保护范围，其溶剂优先选择水、醇或醇-水混合液，这里的醇是指甲醇或乙醇。1,4-benzodiazepine of the present invention Solvates of similar compounds also fall within the protection scope of the present invention. The solvents are preferably water, alcohol or alcohol-water mixtures. The alcohol here is methanol or ethanol.
本发明的第二个目的是提供式(I)所示的1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体、或其与医学上可接受的载体组成的药物组合物在制备ANXA3降解剂中的用途。The second object of the present invention is to provide 1,4-benzodiazepine represented by formula (I) The use of a compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition consisting of the same and a medically acceptable carrier, in the preparation of an ANXA3 degrading agent.
基于表面等离子共振的结合活性测试表明，式(I)所示的1，4-苯二氮类化合物具有亚微摩尔级的ANXA3结合活性，活性结果如表1所示。基于蛋白免疫印迹测试实验表明，所述化合物能够在TNBC细胞中诱导ANXA3蛋白降解，活性具有微摩尔级，结果如表1所示。其中代表性异构体化合物I-9a19、(R)-I-9a19或(S)-I-9a19的降解活性结果如图1所示。The binding activity test based on surface plasmon resonance showed that the 1,4-benzodiazepine represented by formula (I) The compound has submicromolar ANXA3 binding activity, and the activity results are shown in Table 1. Experiments based on Western blot testing showed that the compound can induce ANXA3 protein degradation in TNBC cells, with activity at the micromolar level, and the results are shown in Table 1. The degradation activity results of representative isomer compounds I-9a19, (R)-I-9a19 or (S)-I-9a19 are shown in Figure 1.
本发明的进一步目的是提供式(I)所示的1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体、或其与医学上可接受的载体组成的药物组合物在制备抗肿瘤的药物中的用途，其中所述抗肿瘤药物为预防和/或治疗癌症的药物。通过多种肿瘤细胞的体外抗肿瘤活性测试表明，所述化合物具有微摩尔级的抗肿瘤活性，活性结果如表1所示。尤其是化合物I-9a19能够明显抑制TNBC细胞的克隆、迁移和侵袭等功能，活性结果如图2所示。尤其是化合物I-9a19在体内TNBC移植瘤模型中，显示出有效的治疗效果和诱导肿瘤组织ANXA3蛋白降解作用，活性结果如图3所示。A further object of the present invention is to provide 1,4-benzodiazepine represented by formula (I) The use of a compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition composed thereof and a medically acceptable carrier, in the preparation of an anti-tumor drug, wherein the anti-tumor drug is Medications to prevent and/or treat cancer. In vitro anti-tumor activity tests on various tumor cells show that the compound has anti-tumor activity at the micromolar level, and the activity results are shown in Table 1. In particular, compound I-9a19 can significantly inhibit the cloning, migration and invasion of TNBC cells. The activity results are shown in Figure 2. In particular, compound I-9a19 showed effective therapeutic effects and induced the degradation of ANXA3 protein in tumor tissue in the in vivo TNBC transplanted tumor model. The activity results are shown in Figure 3.
在上述药物中还可以含有一种或多种药学上可接受的载体，所述的载体包括药学领域的常规稀释剂，赋形剂，填充剂、粘合剂，湿润剂，崩解剂，吸收促进剂，表面活性剂，吸附载体，润滑剂等，必要时还可以加入香味剂，甜味剂等。The above-mentioned medicines may also contain one or more pharmaceutically acceptable carriers. The carriers include conventional diluents, excipients, fillers, binders, wetting agents, disintegrating agents, and absorbers in the pharmaceutical field. Accelerators, surfactants, adsorption carriers, lubricants, etc., and flavors, sweeteners, etc. can also be added if necessary.
本发明还提供一种通过作为膜联蛋白ANXA3降解剂来发挥抗肿瘤效果的抗肿瘤药物组合物，所述抗肿瘤药物组合物为片剂、胶囊、丸剂、注射剂、缓控释制剂、喷雾剂或纳米给药体系。The present invention also provides an anti-tumor pharmaceutical composition that exerts an anti-tumor effect by acting as an annexin ANXA3 degrading agent. The anti-tumor pharmaceutical composition is a tablet, capsule, pill, injection, sustained-release preparation, or spray. Or nano drug delivery system.
本发明的有益效果在于所提供的1，4-苯二氮类化合物是一类结构新颖的ANXA3 小分子降解剂，不仅具有亚微摩尔级的ANXA3结合作用，并在TNBC细胞以及动物水平上体现了明显的降解ANXA3活性。而且，在体外能够明显抑制TNBC细胞的增殖、克隆、迁移和侵袭等功能，通过雌性BALB/c nude裸鼠的人源MDA-MB-231细胞原位异种移植瘤模型，证实体内较好发挥抗TNBC的治疗效果，具有良好的潜力和应用前景，可进一步制备成为治疗癌症的药物，所述的癌症为乳腺癌、宫颈癌、卵巢癌、肠癌、胃癌、胰腺癌、肺癌、食管癌、肝癌、白血病和黑色素瘤。优选的，所述抗肿瘤药物为抗三阴性乳腺癌的药物。The beneficial effect of the present invention lies in the 1,4-benzodiazepine provided Compounds are a class of structurally novel ANXA3 The small molecule degrader not only has sub-micromolar ANXA3 binding effect, but also shows obvious ANXA3 degradation activity in TNBC cells and animal levels. Moreover, it can significantly inhibit the proliferation, cloning, migration and invasion of TNBC cells in vitro. Through the orthotopic xenograft tumor model of human MDA-MB-231 cells in female BALB/c nude mice, it has been confirmed that the anti-tumor function is better in vivo. The therapeutic effect of TNBC has good potential and application prospects, and can be further prepared into drugs for the treatment of cancers, such as breast cancer, cervical cancer, ovarian cancer, intestinal cancer, gastric cancer, pancreatic cancer, lung cancer, esophageal cancer, and liver cancer. , leukemia and melanoma. Preferably, the anti-tumor drug is an anti-triple-negative breast cancer drug.
与现有技术相比，本发明的有益效果体现在：Compared with the existing technology, the beneficial effects of the present invention are reflected in:
本发明所提供的1，4-苯二氮类化合物结构及它们在药学上可接受的盐均未见Scifinder报道，是一类结构新颖的ANXA3小分子降解剂。该类化合物通过直接结合ANXA3并诱导其降解作用进而发挥体外和体内抗肿瘤的治疗效果，解决了现有技术中缺乏能够直接结合ANXA3且表现出抗肿瘤活性的小分子药物，为在临床上用于肿瘤的治疗提供了新的治疗选择和用药方案。1,4-benzodiazepine provided by the invention The structures of similar compounds and their pharmaceutically acceptable salts have not been reported by Scifinder. They are a class of ANXA3 small molecule degraders with novel structures. This type of compound exerts anti-tumor therapeutic effects in vitro and in vivo by directly binding to ANXA3 and inducing its degradation. It solves the lack of small molecule drugs that can directly bind to ANXA3 and exhibit anti-tumor activity in the existing technology, and provides a basis for clinical use. It provides new treatment options and medication regimens for the treatment of tumors.
附图说明Description of the drawings
图1：化合物I-9a19、(R)-I-9a19和(S)-I-9a19对ANXA3蛋白水平影响的检测；Figure 1: Detection of the effects of compounds I-9a19, (R)-I-9a19 and (S)-I-9a19 on ANXA3 protein levels;
图2：化合物I-9a19对TNBC细胞克隆、迁移和侵袭等功能影响的检测；Figure 2: Detection of the effect of compound I-9a19 on TNBC cell cloning, migration and invasion;
图3：化合物I-9a19体内抗TNBC药效研究和诱导肿瘤组织ANXA3降解作用检测。Figure 3: In vivo anti-TNBC efficacy study of compound I-9a19 and detection of ANXA3 degradation in tumor tissue.
具体实施方式Detailed ways
下面结合附图和具体实施例对本发明作进一步阐述，但这些实施例只是用于进一步说明本发明，并不改变本发明的保护范围。The present invention will be further described below with reference to the accompanying drawings and specific examples. However, these examples are only used to further illustrate the present invention and do not change the protection scope of the present invention.
实施例1Example 1
2-氧代-2-(3-苯基脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a1)的制备。2-Oxo-2-(3-phenylureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1, 4]Diazepine Preparation of -3-yl)carbodithio (I-9a1).
1.1中间体化合物7的制备
1.1 Preparation of intermediate compound 7
称取5.00g(66.6mmol)氨基甲酸苄酯7.93g(52.5mmol)于100mL圆底烧瓶中，加入甲苯80mL搅拌溶解，逐滴加入7.5mL(66.6mmol)水合乙醛酸，搅拌20min后，将体系升温至80℃，反应2h，抽滤得到化合物1，白色固体16.01g，产率97％；ESI-MS:m/z 349.0[M+Na]+，C16H14N4O4.Weigh 5.00g (66.6mmol) 7.93g (52.5mmol) of benzyl carbamate into a 100mL round-bottomed flask, add 80mL of toluene and stir to dissolve, add 7.5mL (66.6mmol) hydrated glyoxylic acid dropwise, stir for 20 minutes, and The system was heated to 80°C and reacted for 2 hours. Compound 1 was obtained by suction filtration, 16.01g of white solid, yield 97%; ESI-MS: m/z 349.0[M+Na] + , C 16 H 14 N 4 O 4 .
在冰浴条件下，将5.00g(15mmol)化合物1溶于30mL四氢呋喃中冷却至0℃，将2mL(23.6mmol)草酰氯缓慢滴加至该溶液中，后加入2-3滴DMF，反应2h后，将2-氨基苯基苯甲酮3.02g(15.3mmol)和N-甲基吗啉3.67mL(33.7mmol)溶于20mL四氢呋喃中。在0℃下，用恒压滴液漏斗缓慢滴加约20-30min到2中，升温至常温，反应2-4h。将体系用硅藻土过滤后，用20mL四氢呋喃冲洗，将有机相真空浓缩，粗产品用硅胶柱层析纯化(乙酸乙酯/石油醚＝1/2)，得到化合物3，黄色油状物3.8g，产率49％；ESI-MS:m/z 506.0[M+H]+，C29H23N5O4.Under ice bath conditions, 5.00g (15mmol) of compound 1 was dissolved in 30mL of tetrahydrofuran and cooled to 0°C. 2mL (23.6mmol) of oxalyl chloride was slowly added dropwise to the solution, and then 2-3 drops of DMF were added and the reaction was carried out for 2h. Then, 3.02g (15.3mmol) of 2-aminophenylbenzophenone and 3.67mL (33.7mmol) of N-methylmorpholine were dissolved in 20mL of tetrahydrofuran. At 0°C, use a constant pressure dropping funnel to slowly drop into 2 for about 20-30 minutes, raise the temperature to room temperature, and react for 2-4 hours. The system was filtered through diatomaceous earth, rinsed with 20 mL of tetrahydrofuran, and the organic phase was concentrated in vacuo. The crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/2) to obtain compound 3, 3.8 g of yellow oil. , yield 49%; ESI-MS: m/z 506.0[M+H] + , C 29 H 23 N 5 O 4 .
将化合物3 5g(9.9mmol)用10mL甲醇溶解后，置于0℃下，加入15mL氨的甲醇溶液。缓慢升温至室温，反应16h后，过滤掉固体，将有机相真空浓缩。用乙酸乙酯溶解，用1N的氢氧化钠水溶液清洗2次后，用NaCl萃取，有机相用Na2SO4干燥，有机相浓缩后加入60mL乙酸，并加入乙酸铵3.8g(49.3mmol)，在氮气保护下常温反应过夜。反应液真空浓缩后，加入乙酸乙酯：乙醚＝1：3后，加入1N的氢氧化钠水溶液至PH＝8以上，将悬浮液置于0℃下，然后过滤得固体，用水和乙醚连续冲洗固体，得到灰色固体5 1.1g，产率30％；1H NMR(400MHz,DMSO-d6)δ10.87(s,1H),8.47(d,J＝7.6Hz,1H),7.62(s,1H),7.54-7.15(m,13H),5.03(d,J＝16.1Hz,3H).ESI-MS:m/z 386.0[M+H]+,C24H20N2O3.After 5 g (9.9 mmol) of compound 3 was dissolved in 10 mL of methanol, the solution was placed at 0°C, and 15 mL of ammonia methanol solution was added. The temperature was slowly raised to room temperature. After reacting for 16 hours, the solid was filtered off and the organic phase was concentrated in vacuo. Dissolve in ethyl acetate, wash twice with 1N sodium hydroxide aqueous solution, extract with NaCl, dry the organic phase with Na 2 SO 4 , concentrate the organic phase, add 60 mL acetic acid, and add 3.8 g (49.3 mmol) of ammonium acetate. The reaction was carried out overnight at room temperature under nitrogen protection. After the reaction solution was concentrated in vacuum, ethyl acetate: diethyl ether = 1:3 was added, then 1N aqueous sodium hydroxide solution was added until the pH was above 8, the suspension was placed at 0°C, and then filtered to obtain a solid, which was washed continuously with water and diethyl ether. Solid, 1.1g of gray solid 5 was obtained, yield 30%; 1 H NMR (400MHz, DMSO-d 6 ) δ10.87 (s, 1H), 8.47 (d, J = 7.6Hz, 1H), 7.62 (s, 1H),7.54-7.15(m,13H),5.03(d,J＝16.1Hz,3H).ESI-MS:m/z 386.0[M+H] + ,C 24 H 20 N 2 O 3 .
将化合物5 5g(13.0mmol)置于圆底反应瓶中，加入无水碳酸钾3.0g(21.7mmol)和碘甲烷2.5g(17.6mmol)用超干DMF 30mL溶解后，氮气球保护换气3次，常温下反应5h。在0℃下加入冰水和EA淬灭，有机相用饱和食盐水清洗，有机相用Na2SO4干燥，减压后粗产品用硅胶柱层析纯化(乙酸乙酯/石油醚＝1/3)，得到灰色固体6 3g，产率58％；1H NMR(400MHz,DMSO-d6)δ8.51(s,1H),7.71(s,1H),7.64(d,J＝8.0Hz,1H),7.51(s,3H),7.45(d,J＝7.2Hz,2H),7.33(d,J＝19.3Hz,7H),5.04(s,2H),3.35(d,J＝3.7Hz,3H).ESI-MS:m/z 400.2[M+H]+,C24H21N3O3.Place 5g (13.0mmol) of compound 5 in a round-bottomed reaction flask, add 3.0g (21.7mmol) anhydrous potassium carbonate and 2.5g (17.6mmol) methyl iodide, dissolve it in 30mL of ultra-dry DMF, and then use a nitrogen balloon to protect and ventilate 3 times, react at room temperature for 5 hours. Add ice water and EA to quench at 0°C. Wash the organic phase with saturated brine. Dry the organic phase with Na 2 SO 4. After reduced pressure, the crude product is purified by silica gel column chromatography (ethyl acetate/petroleum ether=1/ 3), 3g of gray solid 6 was obtained, with a yield of 58%; 1 H NMR (400MHz, DMSO-d 6 ) δ8.51 (s, 1H), 7.71 (s, 1H), 7.64 (d, J = 8.0Hz, 1H),7.51(s,3H),7.45(d,J＝7.2Hz,2H),7.33(d,J＝19.3Hz,7H),5.04(s,2H),3.35(d,J＝3.7Hz, 3H).ESI-MS:m/z 400.2[M+H] + ,C 24 H 21 N 3 O 3 .
将化合物6 3g(7.5mmol)溶于10mL乙酸，置于圆底反应瓶中，加入30mL 33％溴化氢乙酸溶液，在常温下反应2-4h。体系有固体产生，碳酸氢钠溶液调节至中性，用乙酸乙酯与水萃取，用Na2SO4干燥有机层，粗产品用硅胶柱层析纯化(二氯甲烷/甲醇＝10/1)，得到棕色油状物7 1.4g，产率70％；1H NMR(400MHz,DMSO-d6)δ7.65(t,J＝6.0Hz,1H),7.59(t,J＝6.7Hz,1H),7.46(ddd,J＝20.8,14.2,7.5Hz,5H),7.27(d,J＝5.2Hz,2H),4.26(d,J＝6.0Hz,1H),3.34(s,3H).13C NMR(151MHz,DMSO-d6)δ169.89,164.23,142.76,137.83,131.59,130.18,129.16,129.03,128.21,128.15,123.91,121.74,70.19,34.45.ESI-MS:m/z 266.0[M+H]+，C16H15N3O.Dissolve 3g (7.5mmol) of compound 6 in 10mL acetic acid, place it in a round-bottomed reaction bottle, add 30mL 33% hydrogen bromide acetic acid solution, and react at room temperature for 2-4 hours. When a solid is produced in the system, the sodium bicarbonate solution is adjusted to neutrality, extracted with ethyl acetate and water, the organic layer is dried with Na 2 SO 4 , and the crude product is purified by silica gel column chromatography (dichloromethane/methanol=10/1) , 1.4g of brown oil 7 was obtained, with a yield of 70%; 1 H NMR (400MHz, DMSO-d 6 ) δ7.65 (t, J = 6.0 Hz, 1H), 7.59 (t, J = 6.7 Hz, 1H) 13 C NMR (151MHz, DMSO-d 6 ) δ169.89,164.23,142.76,137.83,131.59,130.18,129.16,129.03,128.21,128.15,123.91,121.74,70.19,34.45.ESI-MS: m/z 266 .0[M+H] + , C 16 H 15 N 3 O.
1.2中间体化合物8a1的制备
1.2 Preparation of intermediate compound 8a1
称取1.70g(18.2mmol)氯乙酰胺于25mL圆底烧瓶中，加入1,2-二氯乙烷20mL搅拌溶解，在0℃下,逐滴加入2mL(23.4mmol)草酰氯，搅拌1h后，将体系升温至90℃，回流5h。再将体系降温至rt，缓缓加入苯胺1.69g(18.2mmol)于上述体系中，搅拌6-8h，抽滤得到白色固体2.83g，产率73％；1H NMR(400MHz,DMSO-d6)δ10.92(s,1H),10.17(s,1H),7.52(d,2H),7.34(t,2H),7.10(t,1H),4.40(s,2H)；13C NMR(151MHz,DMSO-d6)δ168.6,150.2,137.4,128.9,123.8,119.7,43.2；HR-MS(ESI):m/z[M+H]+calcd for C9H9ClN2O2:213.0425,found:213.0425.Weigh 1.70g (18.2mmol) chloroacetamide into a 25mL round-bottomed flask, add 20mL of 1,2-dichloroethane and stir to dissolve. At 0°C, add 2mL (23.4mmol) oxalyl chloride dropwise, and stir for 1 hour. , the system was heated to 90°C and refluxed for 5 hours. Then cool the system to rt, slowly add 1.69g (18.2mmol) of aniline to the above system, stir for 6-8h, and obtain 2.83g of white solid by suction filtration, with a yield of 73%; 1 H NMR (400MHz, DMSO-d 6 )δ10.92(s,1H),10.17(s,1H),7.52(d,2H),7.34(t,2H),7.10(t,1H),4.40(s,2H); 13 C NMR(151MHz ,DMSO-d 6 )δ168.6,150.2,137.4,128.9,123.8,119.7,43.2; HR-MS(ESI):m/z[M+H] + calcd for C 9 H 9 ClN 2 O 2 :213.0425, found :213.0425.
1.3终产物I-9a1的制备
1.3 Preparation of final product I-9a1
取一个25mL的圆底烧瓶，加入200mg(0.75mmol)化合物7，210μL三乙胺和110μL的CS2，室温搅拌2h。随后加入6mL乙腈溶解体系，再加入160mg(0.75mmol)化合物8a1，室温反应6h后TLC板监测反应(取少量反应液，加入饱和氯化钠溶液和乙酸乙酯，取乙酸乙酯层点板)，原料点基本消失，停止反应，将体系蒸干，乙酸乙酯与水萃取，有机相用无水硫酸钠干燥后旋干溶剂，粗产品用硅胶柱层析纯化(乙酸乙酯/石油醚＝2/3)，得到黄色固体130mg，产率33％；1H NMR(400MHz,DMSO-d6)δ11.55(s,1H),10.96(s,1H),10.33(s,1H),7.92-6.95(m,14H),5.97(s,1H),4.25(s,2H),3.39(s,3H)；13C NMR(150MHz,DMSO-d6)δ196.47,170.13,166.95,165.92,150.27,142.53,137.32,137.23,132.29,130.75,129.59,129.34,128.79,128.24,127.87,124.41,123.55,122.07,119.47,75.31,38.58,34.73；ESI-MS:m/z 517.8[M+H]+,C26H23N5O3S2,HRMS calcd.518.1315[M+H]+,found 518.1319.Take a 25 mL round-bottomed flask, add 200 mg (0.75 mmol) compound 7, 210 μL triethylamine and 110 μL CS 2 and stir at room temperature for 2 hours. Then add 6 mL of acetonitrile to dissolve the system, and then add 160 mg (0.75 mmol) of compound 8a1. After reacting at room temperature for 6 hours, monitor the reaction on a TLC plate (take a small amount of the reaction solution, add saturated sodium chloride solution and ethyl acetate, and take an ethyl acetate layer plate) , the raw material point basically disappears, the reaction is stopped, the system is evaporated to dryness, extracted with ethyl acetate and water, the organic phase is dried with anhydrous sodium sulfate and then the solvent is spun dry, the crude product is purified by silica gel column chromatography (ethyl acetate/petroleum ether= 2/3), obtained 130 mg of yellow solid, yield 33%; 1 H NMR (400MHz, DMSO-d 6 ) δ11.55 (s, 1H), 10.96 (s, 1H), 10.33 (s, 1H), 7.92 -6.95(m,14H),5.97(s,1H),4.25(s,2H),3.39(s,3H); 13 C NMR(150MHz,DMSO-d 6 )δ196.47,170.13,166.95,165.92,150.27, 142.53,137.32,137.23,132.29,130.75,129.59,129.34,128.79,128.24,127.87,124.41,123.55,122.07,119.47,75.31,38.58,34.73; ESI-MS:m/ z 517.8[M+H] + ,C 26 H 23 N 5 O 3 S 2 ,HRMS calcd.518.1315[M+H] + ,found 518.1319.
实施例2Example 2
2-氧代-2-(3-(4-三氟甲基)苯基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a2)的制备2-Oxo-2-(3-(4-trifluoromethyl)phenyl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H -Benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a2)
2.1中间体2-氯-N-(4-(三氟甲基)苯基)氨甲酰)乙酰胺(8a2)的制备2.1 Preparation of intermediate 2-chloro-N-(4-(trifluoromethyl)phenyl)carbamoyl)acetamide (8a2)
按照实施例1中步骤1.2的方法合成中间体8a2。1H NMR(400MHz,DMSO-d6)δ11.14-10.96(m,1H),10.41(s,1H),7.81-7.68(m,4H),4.43(s,2H).Intermediate 8a2 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.14-10.96(m,1H),10.41(s,1H),7.81-7.68(m,4H),4.43(s,2H).
2.2终产物I-9a2的制备2.2 Preparation of final product I-9a2
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a2，制得化合物I-9a2。1H NMR(400MHz,DMSO-d6)δ11.50(s,1H),11.04(s,1H),10.54(s,1H),7.60(dd,J＝71.2,29.4Hz,13H),5.95(s,1H),4.24(s,2H),3.36(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a2, and prepare compound I-9a2. 1 H NMR (400MHz, DMSO-d 6 ) δ11.50 (s, 1H), 11.04 (s, 1H), 10.54 (s, 1H), 7.60 (dd, J = 71.2, 29.4Hz, 13H), 5.95 ( s,1H),4.24(s,2H),3.36(s,3H) .
实施例3Example 3
2-氧代-2-(3-(2-(三氟甲基)苯基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a3)的制备2-Oxo-2-(3-(2-(trifluoromethyl)phenyl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro- 1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a3)
3.1中间体2-氯-N-(2-(三氟甲基)苯基)氨甲酰)乙酰胺(8a3)的制备3.1 Preparation of intermediate 2-chloro-N-(2-(trifluoromethyl)phenyl)carbamoyl)acetamide (8a3)
按照实施例1中步骤1.2的方法合成中间体8a3。1H NMR(400MHz,DMSO-d6)δ11.23(s,1H),10.60(s,1H),8.07(d,J＝8.2Hz,1H),7.77-7.64(m,2H),7.35(d,J＝7.5Hz,1H),4.39(d,J＝7.3Hz,2H).Intermediate 8a3 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.23 (s, 1H), 10.60 (s, 1H), 8.07 (d, J = 8.2Hz, 1H), 7.77-7.64 (m, 2H), 7.35 ( d,J＝7.5Hz,1H),4.39(d,J＝7.3Hz,2H).
3.2终产物9a3的制备3.2 Preparation of final product 9a3
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a3，制得化合物I-9a3。1H NMR(400MHz,DMSO-d6)δ11.53(s,1H),11.20(s,1H),10.73(s,1H),8.09(s,1H),7.68(s,5H),7.59-7.43(m,5H),7.30(s,2H),5.94(s,1H),4.25(s,2H),3.37(s,3H). Compound I-9a3 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8a3. 1 H NMR (400MHz, DMSO-d 6 ) δ11.53(s,1H),11.20(s,1H),10.73(s,1H),8.09(s,1H),7.68(s,5H),7.59- 7.43(m,5H),7.30(s,2H),5.94(s,1H),4.25(s,2H),3.37(s,3H) .
实施例4Example 4
2-(3-(4-(二甲氨基)苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a4)的制备2-(3-(4-(Dimethylamino)phenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H -Benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a4)
4.1中间体2-氯-N-(4-(二甲氨基)苯基)氨甲酰)乙酰胺(8a4)的制备4.1 Preparation of intermediate 2-chloro-N-(4-(dimethylamino)phenyl)carbamoyl)acetamide (8a4)
按照实施例1中步骤1.2的方法合成中间体8a4。1H NMR(400MHz,DMSO-d6)δ10.87-10.73(m,1H),9.96-9.80(m,1H),7.29(s,2H),6.68(s,2H),4.33(s,2H),2.83(d,J＝3.3Hz,6H).Intermediate 8a4 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.87-10.73(m,1H),9.96-9.80(m,1H),7.29(s,2H),6.68(s,2H),4.33(s,2H ),2.83(d,J＝3.3Hz,6H).
4.2终产物I-9a4的制备4.2 Preparation of final product I-9a4
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a4，制得化合物I-9a4。1H NMR(400MHz,DMSO-d6)δ11.46(d,J＝6.3Hz,1H),10.75(s,1H),9.99(s,1H),7.66(s,2H),7.53(s,3H),7.44(d,J＝7.2Hz,2H),7.25(d,J＝9.8Hz,4H),6.64(s,2H),5.92(s,1H),4.17(s,2H),3.33(s,3H),2.79(s,6H). Synthesize according to the method in step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a4, and prepare compound I-9a4. 1 H NMR (400MHz, DMSO-d 6 ) δ11.46 (d, J = 6.3 Hz, 1H), 10.75 (s, 1H), 9.99 (s, 1H), 7.66 (s, 2H), 7.53 (s, 3H),7.44(d,J＝7.2Hz,2H),7.25(d,J＝9.8Hz,4H),6.64(s,2H),5.92(s,1H),4.17(s,2H),3.33( s,3H),2.79(s,6H) .
实施例5Example 5
2-氧代-2-(3-(4-(三氟甲氧基)苯基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a5)的制备2-Oxo-2-(3-(4-(trifluoromethoxy)phenyl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a5)
5.1中间体2-氯-N-((4-(三氟甲氧基)苯基)氨甲酰)乙酰胺(8a5)的制备5.1 Preparation of intermediate 2-chloro-N-((4-(trifluoromethoxy)phenyl)carbamoyl)acetamide (8a5)
按照实施例1中步骤1.2的方法合成中间体8a5。1H NMR(400MHz,DMSO-d6)δ10.97(s,1H),10.24(s,1H),7.66(td,J＝6.1,3.0Hz,2H),7.35(t,J＝6.8Hz,2H),4.47-4.35(m,2H).Intermediate 8a5 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.97 (s, 1H), 10.24 (s, 1H), 7.66 (td, J = 6.1, 3.0Hz, 2H), 7.35 (t, J = 6.8Hz, 2H),4.47-4.35(m,2H).
5.2终产物I-9a5的制备5.2 Preparation of final product I-9a5
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a5，制得化合物I-9a5。1H NMR(400MHz,DMSO-d6)δ11.50(d,J＝6.7Hz,1H),10.97(s,1H),10.39(s,1H),7.75-7.68(m,2H),7.65-7.61(m,2H),7.60-7.57(m,2H),7.55-7.52(m,1H),7.47(t,J＝7.5Hz,2H),7.31(dp,J＝7.8,2.0Hz,4H),5.96(d,J＝6.7Hz,1H),4.25(s,2H),3.38(d,J＝1.6Hz,3H). Compound I-9a5 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8a5. 1 H NMR (400MHz, DMSO-d 6 ) δ11.50 (d, J = 6.7Hz, 1H), 10.97 (s, 1H), 10.39 (s, 1H), 7.75-7.68 (m, 2H), 7.65- 7.61(m,2H),7.60-7.57(m,2H),7.55-7.52(m,1H),7.47(t,J＝7.5Hz,2H),7.31(dp,J＝7.8,2.0Hz,4H) ,5.96(d,J＝6.7Hz,1H),4.25(s,2H),3.38(d,J＝1.6Hz,3H) .
实施例6 Example 6
2-(3-(3-氟苯基)脲基)-2-氧代乙基(1-甲基-2-氧基-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a6)的制备2-(3-(3-Fluorophenyl)ureido)-2-oxoethyl(1-methyl-2-oxy-5-phenyl-2,3-dihydro-1H-benzo[ e][1,4]diaza Preparation of -3-yl)carbodithio(I-9a6)
6.1中间体2-氯-N-((3-氟苯基)氨甲酰)乙酰胺(8a6)的制备6.1 Preparation of intermediate 2-chloro-N-((3-fluorophenyl)carbamoyl)acetamide (8a6)
按照实施例1中步骤1.2的方法合成中间体8a6。1H NMR(400MHz,DMSO-d6)δ10.97(s,1H),10.27(s,1H),7.63-7.52(m,1H),7.44-7.33(m,1H),7.33-7.26(m,1H),7.04-6.89(m,1H),4.39(s,2H).Intermediate 8a6 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.97(s,1H),10.27(s,1H),7.63-7.52(m,1H),7.44-7.33(m,1H),7.33-7.26(m ,1H),7.04-6.89(m,1H),4.39(s,2H).
6.2终产物I-9a6的制备6.2 Preparation of final product I-9a6
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a6，制得化合物I-9a6。1H NMR(400MHz,DMSO-d6)δ11.49(s,1H),10.92(d,J＝54.2Hz,1H),10.45(s,1H),7.71(d,J＝10.2Hz,2H),7.64-7.56(m,2H),7.51(dd,J＝22.4,6.1Hz,4H),7.37-7.29(m,3H),7.23(d,J＝8.5Hz,1H),6.90(s,1H),5.96(s,1H),4.26(s,2H),3.35(s,3H). Compound I-9a6 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8a6. 1 H NMR (400MHz, DMSO-d 6 ) δ11.49 (s, 1H), 10.92 (d, J = 54.2Hz, 1H), 10.45 (s, 1H), 7.71 (d, J = 10.2Hz, 2H) ,7.64-7.56(m,2H),7.51(dd,J＝22.4,6.1Hz,4H),7.37-7.29(m,3H),7.23(d,J＝8.5Hz,1H),6.90(s,1H ),5.96(s,1H),4.26(s,2H),3.35(s,3H) .
实施例7Example 7
2-(3-(3-氯苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a7)的制备2-(3-(3-chlorophenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[ e][1,4]diaza Preparation of -3-yl)carbodithio(I-9a7)
7.1中间体2-氯-N-((3-氯苯基)氨甲酰)乙酰胺(8a7)的制备7.1 Preparation of intermediate 2-chloro-N-((3-chlorophenyl)carbamoyl)acetamide (8a7)
按照实施例1中步骤1.2的方法合成中间体8a7。ESI-MS:m/z 247.0[M+H]+。Intermediate 8a7 was synthesized according to the method of step 1.2 in Example 1. ESI-MS: m/z 247.0[M+H] + .
7.2终产物I-9a7的制备7.2 Preparation of final product I-9a7
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a7，制得化合物I-9a7。1H NMR(400MHz,DMSO-d6)δ11.50(s,1H),11.00(s,1H),10.41(s,1H),7.72(dd,J＝22.1,11.6Hz,3H),7.63-7.52(m,3H),7.48(t,J＝7.4Hz,2H),7.35(d,J＝16.8Hz,4H),7.14(d,J＝7.5Hz,1H),5.97(s,1H),4.26(s,2H),3.39(s,3H). Compound I-9a7 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8a7. 1 H NMR (400MHz, DMSO-d 6 ) δ11.50 (s, 1H), 11.00 (s, 1H), 10.41 (s, 1H), 7.72 (dd, J = 22.1, 11.6Hz, 3H), 7.63- 7.52(m,3H),7.48(t,J＝7.4Hz,2H),7.35(d,J＝16.8Hz,4H),7.14(d,J＝7.5Hz,1H),5.97(s,1H), 4.26(s,2H),3.39(s,3H) .
实施例8Example 8
2-(3-(4-溴苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a8)的制备2-(3-(4-bromophenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[ e][1,4]diaza Preparation of -3-yl)carbodithio(I-9a8)
8.1中间体N-((4-溴苯基)氨甲酰)-2-氯乙酰胺(8a8)的制备8.1 Preparation of intermediate N-((4-bromophenyl)carbamoyl)-2-chloroacetamide (8a8)
按照实施例1中步骤1.2的方法合成中间体8a8。1H NMR(400MHz,DMSO-d6)δ10.99(d,J＝28.7Hz,1H),10.19(s,1H),7.54-7.49(m,4H),4.39(d,J＝4.5Hz,2H).Intermediate 8a8 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.99 (d, J = 28.7Hz, 1H), 10.19 (s, 1H), 7.54-7.49 (m, 4H), 4.39 (d, J = 4.5Hz, 2H).
8.2终产物I-9a8的制备8.2 Preparation of final product I-9a8
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a8，制得化合物I-9a8。1H NMR(400MHz,DMSO-d6)δ11.49(d,J＝8.8Hz,1H),10.96(s,1H),10.34(s,1H),7.76-7.67(m,2H),7.59(d,J＝8.0Hz,2H),7.50(d,J＝7.7Hz,7H),7.37-7.29(m,2H),5.96(d,J＝7.8Hz,1H),4.24(s,2H),3.38(s,3H). Compound I-9a8 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8a8. 1 H NMR (400MHz, DMSO-d 6 ) δ11.49 (d, J = 8.8 Hz, 1H), 10.96 (s, 1H), 10.34 (s, 1H), 7.76-7.67 (m, 2H), 7.59 ( d,J＝8.0Hz,2H),7.50(d,J＝7.7Hz,7H),7.37-7.29(m,2H),5.96(d,J＝7.8Hz,1H),4.24(s,2H), 3.38(s,3H) .
实施例9Example 9
2-(3-(3-溴苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a9)的制备2-(3-(3-bromophenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[ e][1,4]diaza Preparation of -3-yl)carbodithio(I-9a9)
9.1中间体N-((3-溴苯基)氨甲酰)-2-氯乙酰胺(8a9)的制备 9.1 Preparation of intermediate N-((3-bromophenyl)carbamoyl)-2-chloroacetamide (8a9)
按照实施例1中步骤1.2的方法合成中间体8a9。1H NMR(400MHz,DMSO-d6)δ10.99(s,1H),10.23(s,1H),7.92(s,1H),7.45(d,J＝6.0Hz,1H),7.30(d,J＝4.6Hz,2H),4.43-4.39(m,2H).Intermediate 8a9 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.99 (s, 1H), 10.23 (s, 1H), 7.92 (s, 1H), 7.45 (d, J = 6.0Hz, 1H), 7.30 (d, J＝4.6Hz,2H),4.43-4.39(m,2H).
9.2终产物I-9a9的制备9.2 Preparation of final product I-9a9
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a9，制得化合物I-9a9。1H NMR(400MHz,DMSO-d6)δ11.56-11.47(m,1H),11.01(s,1H),10.39(s,1H),7.91(s,1H),7.73(dd,J＝14.2,6.6Hz,3H),7.60(d,J＝7.9Hz,2H),7.55(d,J＝7.0Hz,1H),7.50(d,J＝7.4Hz,2H),7.41(s,1H),7.34(d,J＝7.5Hz,2H),7.27(d,J＝4.5Hz,2H),5.98(d,J＝6.6Hz,1H),4.26(s,2H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a9, and prepare compound I-9a9. 1 H NMR (400MHz, DMSO-d 6 ) δ11.56-11.47(m,1H),11.01(s,1H),10.39(s,1H),7.91(s,1H),7.73(dd,J＝14.2 ,6.6Hz,3H),7.60(d,J＝7.9Hz,2H),7.55(d,J＝7.0Hz,1H),7.50(d,J＝7.4Hz,2H),7.41(s,1H), 7.34(d,J＝7.5Hz,2H),7.27(d,J＝4.5Hz,2H),5.98(d,J＝6.6Hz,1H),4.26(s,2H) .
实施例10Example 10
2-(3-(2-氰基苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a10)的制备2-(3-(2-cyanophenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo [e][1,4]diazapine Preparation of -3-yl)carbodithio(I-9a10)
10.1中间体2-氯-N-((2-氰基苯基)氨甲酰)乙酰胺(8a10)的制备10.1 Preparation of intermediate 2-chloro-N-((2-cyanophenyl)carbamoyl)acetamide (8a10)
按照实施例1中步骤1.2的方法合成中间体8a10。1H NMR(400MHz,DMSO-d6)δ11.30(s,1H),10.81(s,1H),8.24-8.12(m,1H),7.86(t,J＝6.1Hz,1H),7.73(q,J＝6.9Hz,1H),7.32(q,J＝7.3,6.6Hz,1H),4.43(d,J＝4.7Hz,2H).Intermediate 8a10 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.30 (s, 1H), 10.81 (s, 1H), 8.24-8.12 (m, 1H), 7.86 (t, J = 6.1Hz, 1H), 7.73 ( q, J＝6.9Hz, 1H), 7.32 (q, J＝7.3, 6.6Hz, 1H), 4.43 (d, J＝4.7Hz, 2H).
10.2终产物I-9a10的制备10.2 Preparation of final product I-9a10
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a10，制得化合物I-9a10。1H NMR(400MHz,DMSO-d6)δ11.57(d,J＝6.8Hz,1H),11.28(s,1H),10.97(s,1H),8.19(d,J＝8.5Hz,1H),7.83(d,J＝8.0Hz,1H),7.72(s,3H),7.61(d,J＝7.2Hz,2H),7.56(d,J＝6.1Hz,1H),7.50(d,J＝7.4Hz,2H),7.33(t,J＝13.8Hz,3H),5.98(d,J＝6.5Hz,1H),4.29(s,2H),3.40(s,3H).Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a10, and prepare compound I-9a10. 1 H NMR (400MHz, DMSO-d 6 ) δ11.57(d,J＝6.8Hz,1H),11.28(s,1H),10.97(s,1H),8.19(d,J＝8.5Hz,1H) ,7.83(d,J＝8.0Hz,1H),7.72(s,3H),7.61(d,J＝7.2Hz,2H),7.56(d,J＝6.1Hz,1H),7.50(d,J＝ 7.4Hz,2H),7.33(t,J＝13.8Hz,3H),5.98(d,J＝6.5Hz,1H),4.29(s,2H),3.40(s,3H).
实施例11Example 11
2-(3-(2-乙酰基苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a11)的制备2-(3-(2-acetylphenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo [e][1,4]diazapine Preparation of -3-yl)carbodithio(I-9a11)
11.1中间体N-((2-乙酰基苯基)氨甲酰)-2-氯乙酰胺(8a11)的制备11.1 Preparation of intermediate N-((2-acetylphenyl)carbamoyl)-2-chloroacetamide (8a11)
按照实施例1中步骤1.2的方法合成中间体8a11。1H NMR(400MHz,DMSO-d6)δ12.13(s,1H),10.93(s,1H),8.38(d,J＝8.5Hz,1H),8.04(d,J＝7.9Hz,1H),7.61(t,J＝8.0Hz,1H),7.24(t,J＝7.6Hz,1H),4.37(s,2H),2.63(s,3H).Intermediate 8a11 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ12.13 (s, 1H), 10.93 (s, 1H), 8.38 (d, J = 8.5Hz, 1H), 8.04 (d, J = 7.9Hz, 1H) ,7.61(t,J＝8.0Hz,1H),7.24(t,J＝7.6Hz,1H),4.37(s,2H),2.63(s,3H).
11.2终产物I-9a11的制备11.2 Preparation of final product I-9a11
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a11，制得化合物I-9a11。1H NMR(400MHz,DMSO-d6)δ12.16(s,1H),11.54(d,J＝6.3Hz,1H),10.88(s,1H),8.38(d,J＝8.5Hz,1H),8.00(d,J＝8.1Hz,1H),7.72(d,J＝10.1Hz,2H),7.61(d,J＝7.7Hz,2H),7.57(d,J＝8.0Hz,2H),7.48(t,J＝7.5Hz,2H),7.34(d,J＝8.1Hz,2H),7.22(d,J＝8.0Hz,1H),6.00-5.96(m,1H),4.23(d,J＝3.9Hz,2H),3.40(s,3H),2.60(s,3H).Compound I-9a11 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8a11. 1 H NMR (400MHz, DMSO-d 6 ) δ12.16 (s, 1H), 11.54 (d, J = 6.3Hz, 1H), 10.88 (s, 1H), 8.38 (d, J = 8.5Hz, 1H) ,8.00(d,J＝8.1Hz,1H),7.72(d,J＝10.1Hz,2H),7.61(d,J＝7.7Hz,2H),7.57(d,J＝8.0Hz,2H),7.48 (t,J＝7.5Hz,2H),7.34(d,J＝8.1Hz,2H),7.22(d,J＝8.0Hz,1H),6.00-5.96(m,1H),4.23(d,J＝ 3.9Hz,2H),3.40(s,3H),2.60(s,3H).
实施例12 Example 12
2-(3-(3-乙基苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a12)的制备2-(3-(3-ethylphenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo [e][1,4]diazapine Preparation of -3-yl)carbodithio(I-9a12)
12.1中间体2-氯-N-((3-乙基苯基)氨甲酰)乙酰胺(8a12)的制备12.1 Preparation of intermediate 2-chloro-N-((3-ethylphenyl)carbamoyl)acetamide (8a12)
按照实施例1中步骤1.2的方法合成中间体8a12。1H NMR(400MHz,DMSO-d6)δ10.89(s,1H),10.13(s,1H),7.35(d,J＝7.3Hz,2H),7.24(t,J＝7.9Hz,1H),6.96(d,J＝7.5Hz,1H),4.40(s,2H),2.59(q,J＝7.6Hz,2H),1.18(t,J＝7.6Hz,3H).Intermediate 8a12 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.89 (s, 1H), 10.13 (s, 1H), 7.35 (d, J = 7.3Hz, 2H), 7.24 (t, J = 7.9Hz, 1H) ,6.96(d,J＝7.5Hz,1H),4.40(s,2H),2.59(q,J＝7.6Hz,2H),1.18(t,J＝7.6Hz,3H).
12.2终产物I-9a12的制备12.2 Preparation of final product I-9a12
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a12，制得化合物I-9a12。1H NMR(400MHz,DMSO-d6)δ11.56(s,1H),10.96(s,1H),10.34(s,1H),7.74(d,J＝10.6Hz,2H),7.62(d,J＝7.6Hz,2H),7.57(d,J＝6.6Hz,1H),7.51(d,J＝7.2Hz,2H),7.35(s,4H),7.24(s,1H),6.95(d,J＝7.3Hz,1H),5.99(s,1H),4.27(s,2H),3.62(s,3H),1.78(s,2H),1.19(d,J＝7.8Hz,3H).Synthesize according to the method in step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a12, and prepare compound I-9a12. 1 H NMR (400MHz, DMSO-d 6 ) δ11.56 (s, 1H), 10.96 (s, 1H), 10.34 (s, 1H), 7.74 (d, J = 10.6Hz, 2H), 7.62 (d, J＝7.6Hz,2H),7.57(d,J＝6.6Hz,1H),7.51(d,J＝7.2Hz,2H),7.35(s,4H),7.24(s,1H),6.95(d, J＝7.3Hz,1H),5.99(s,1H),4.27(s,2H),3.62(s,3H),1.78(s,2H),1.19(d,J＝7.8Hz,3H).
实施例13Example 13
2-(3-(5-氯-2-(三氟甲基)苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a13)的制备2-(3-(5-chloro-2-(trifluoromethyl)phenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3 -Dihydro-1H-benzo[e][1,4]diaza Preparation of -3-yl)carbodithio(I-9a13)
13.1中间体2-氯-N-((5-氯-2-(三氟甲基)苯基)氨甲酰)乙酰胺(8a13)的制备13.1 Preparation of intermediate 2-chloro-N-((5-chloro-2-(trifluoromethyl)phenyl)carbamoyl)acetamide (8a13)
按照实施例1中步骤1.2的方法合成中间体8a13。1H NMR(400MHz,DMSO-d6)δ11.38(s,1H),10.80(s,1H),8.26(s,1H),7.77(d,J＝8.5Hz,1H),7.43(d,J＝8.6Hz,1H),4.40(d,J＝1.9Hz,2H).Intermediate 8a13 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.38 (s, 1H), 10.80 (s, 1H), 8.26 (s, 1H), 7.77 (d, J = 8.5Hz, 1H), 7.43 (d, J＝8.6Hz,1H),4.40(d,J＝1.9Hz,2H).
13.2终产物I-9a13的制备13.2 Preparation of final product I-9a13
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a13，制得化合物I-9a13。1H NMR(400MHz,DMSO-d6)δ11.52(s,1H),11.32(s,1H),10.91(s,1H),8.25(s,1H),7.71(d,J＝9.4Hz,2H),7.59-7.51(m,3H),7.45(d,J＝7.5Hz,2H),7.37(d,J＝8.4Hz,1H),7.29(d,J＝8.7Hz,2H),5.92(s,1H),4.23(s,2H),3.35(s,3H). Synthesize according to the method in step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a13, and prepare compound I-9a13. 1 H NMR (400MHz, DMSO-d 6 ) δ11.52 (s, 1H), 11.32 (s, 1H), 10.91 (s, 1H), 8.25 (s, 1H), 7.71 (d, J = 9.4Hz, 2H),7.59-7.51(m,3H),7.45(d,J＝7.5Hz,2H),7.37(d,J＝8.4Hz,1H),7.29(d,J＝8.7Hz,2H),5.92( s,1H),4.23(s,2H),3.35(s,3H) .
实施例14Example 14
2-(3-(3-氯-5-硝基苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a14)的制备2-(3-(3-chloro-5-nitrophenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro- 1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a14)
14.1中间体2-氯-N-((3-氯-5-硝基苯基)氨甲酰)乙酰胺(8a14)的制备14.1 Preparation of intermediate 2-chloro-N-((3-chloro-5-nitrophenyl)carbamoyl)acetamide (8a14)
按照实施例1中步骤1.2的方法合成中间体8a14。1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),10.49(s,1H),8.54(q,J＝1.9Hz,1H),8.10(d,J＝2.1Hz,1H),7.99(q,J＝1.8Hz,1H),4.42(d,J＝1.6Hz,2H).Intermediate 8a14 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.12(s,1H),10.49(s,1H),8.54(q,J＝1.9Hz,1H),8.10(d,J＝2.1Hz,1H) ,7.99(q,J＝1.8Hz,1H),4.42(d,J＝1.6Hz,2H).
14.2终产物I-9a14的制备14.2 Preparation of final product I-9a14
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a14，制得化合物I-9a14。1H NMR(400MHz,DMSO-d6)δ11.50(s,1H),11.15(s,1H),10.66(s,1H),8.54(s,1H),8.10(s,1H),7.95(s,1H),7.74-7.67(m,2H),7.59(d,J＝6.4Hz,2H),7.54(d,J＝8.1Hz,1H),7.52-7.45(m,2H),7.33(d,J＝6.3Hz,2H),5.97(d,J＝8.0Hz,1H),4.27(s,2H),3.38(s, 3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a14, and prepare compound I-9a14. 1 H NMR (400MHz, DMSO-d 6 ) δ11.50(s,1H),11.15(s,1H),10.66(s,1H),8.54(s,1H),8.10(s,1H),7.95( s,1H),7.74-7.67(m,2H),7.59(d,J＝6.4Hz,2H),7.54(d,J＝8.1Hz,1H),7.52-7.45(m,2H),7.33(d ,J＝6.3Hz,2H),5.97(d,J＝8.0Hz,1H),4.27(s,2H),3.38(s, 3H) .
实施例15Example 15
2-(3-(5-氟-2-硝基苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a15)的制备2-(3-(5-fluoro-2-nitrophenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro- 1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a15)
15.1中间体2-氯-N-((5-氟-2-硝基苯基)氨甲酰)乙酰胺(8a15)的制备15.1 Preparation of intermediate 2-chloro-N-((5-fluoro-2-nitrophenyl)carbamoyl)acetamide (8a15)
按照实施例1中步骤1.2的方法合成中间体8a15。1H NMR(400MHz,DMSO-d6)δ12.07(s,1H),11.35(s,1H),8.33(ddd,J＝15.1,10.6,4.4Hz,2H),7.21(td,J＝6.9,3.6Hz,1H),4.39(d,J＝5.5Hz,2H).Intermediate 8a15 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ12.07 (s, 1H), 11.35 (s, 1H), 8.33 (ddd, J = 15.1, 10.6, 4.4Hz, 2H), 7.21 (td, J = 6.9 ,3.6Hz,1H),4.39(d,J=5.5Hz,2H).
15.2终产物I-9a15的制备15.2 Preparation of final product I-9a15
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a15，制得化合物I-9a15。1H NMR(400MHz,DMSO-d6)δ12.16(s,1H),11.52(d,J＝5.5Hz,1H),11.29(s,1H),8.42-8.24(m,2H),7.76-7.65(m,2H),7.59(d,J＝6.3Hz,2H),7.53(d,J＝8.2Hz,1H),7.52-7.45(m,2H),7.32(d,J＝5.6Hz,2H),7.17(t,J＝8.3Hz,1H),5.96(d,J＝5.8Hz,1H),4.26(s,2H),3.38(s,3H). Synthesize according to the method in step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a15, and prepare compound I-9a15. 1 H NMR (400MHz, DMSO-d 6 ) δ12.16 (s, 1H), 11.52 (d, J = 5.5Hz, 1H), 11.29 (s, 1H), 8.42-8.24 (m, 2H), 7.76- 7.65(m,2H),7.59(d,J＝6.3Hz,2H),7.53(d,J＝8.2Hz,1H),7.52-7.45(m,2H),7.32(d,J＝5.6Hz,2H ),7.17(t,J＝8.3Hz,1H),5.96(d,J＝5.8Hz,1H),4.26(s,2H),3.38(s,3H) .
实施例16Example 16
2-(3-(3-氯-4-硝基苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a16)的制备2-(3-(3-chloro-4-nitrophenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro- 1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a16)
16.1中间体2-氯-N-((3-氯-4-硝基苯基)氨甲酰)乙酰胺(8a16)的制备16.1 Preparation of intermediate 2-chloro-N-((3-chloro-4-nitrophenyl)carbamoyl)acetamide (8a16)
按照实施例1中步骤1.2的方法合成中间体8a16。1H NMR(400MHz,DMSO-d6)δ11.13(s,1H),10.53(s,1H),8.12(dd,J＝8.9,4.1Hz,1H),8.04(d,J＝3.5Hz,1H),7.71(d,J＝9.0Hz,1H),4.43(d,J＝4.0Hz,2H).Intermediate 8a16 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.13 (s, 1H), 10.53 (s, 1H), 8.12 (dd, J = 8.9, 4.1Hz, 1H), 8.04 (d, J = 3.5Hz, 1H), 7.71 (d, J = 9.0Hz, 1H), 4.43 (d, J = 4.0Hz, 2H).
16.2终产物I-9a16的制备16.2 Preparation of final product I-9a16
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a16，制得化合物I-9a16。1H NMR(400MHz,DMSO-d6)δ11.51(d,J＝7.2Hz,1H),11.17(s,1H),10.69(s,1H),8.09(d,J＝9.2Hz,1H),8.03(s,1H),7.70(q,J＝6.7Hz,3H),7.58(d,J＝8.2Hz,2H),7.54(d,J＝5.6Hz,1H),7.47(t,J＝8.2Hz,2H),7.38-7.27(m,3H),5.96(d,J＝8.6Hz,1H),4.27(s,2H),3.38(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a16, and prepare compound I-9a16. 1 H NMR (400MHz, DMSO-d 6 ) δ11.51 (d, J = 7.2 Hz, 1H), 11.17 (s, 1H), 10.69 (s, 1H), 8.09 (d, J = 9.2 Hz, 1H) ,8.03(s,1H),7.70(q,J＝6.7Hz,3H),7.58(d,J＝8.2Hz,2H),7.54(d,J＝5.6Hz,1H),7.47(t,J＝ 8.2Hz,2H),7.38-7.27(m,3H),5.96(d,J=8.6Hz,1H),4.27(s,2H),3.38(s,3H) .
实施例17Example 17
2-(3-(5-氯-2-甲氧基苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a17)的制备2-(3-(5-chloro-2-methoxyphenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a17)
17.1中间体2-氯-N-((5-氯-2-甲氧基苯基)氨甲酰)乙酰胺(8a17)的制备17.1 Preparation of intermediate 2-chloro-N-((5-chloro-2-methoxyphenyl)carbamoyl)acetamide (8a17)
按照实施例1中步骤1.2的方法合成中间体8a17。1H NMR(400MHz,DMSO-d6)δ11.18(d,J＝14.0Hz,1H),10.74(s,1H),8.33-8.13(m,1H),7.25-7.08(m,2H),4.45-4.34(m,2H),3.91(dd,J＝13.7,3.4Hz,3H).Intermediate 8a17 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.18 (d, J = 14.0Hz, 1H), 10.74 (s, 1H), 8.33-8.13 (m, 1H), 7.25-7.08 (m, 2H), 4.45-4.34(m,2H),3.91(dd,J=13.7,3.4Hz,3H).
17.2终产物I-9a17的制备 17.2 Preparation of final product I-9a17
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a17，制得化合物I-9a17。1H NMR(400MHz,DMSO-d6)δ11.56(d,J＝6.7Hz,1H),11.14(s,1H),10.85(s,1H),8.22(s,1H),7.76-7.68(m,2H),7.60(d,J＝7.8Hz,2H),7.55(d,J＝7.0Hz,1H),7.49(t,J＝7.5Hz,2H),7.34(d,J＝8.3Hz,2H),7.09(d,J＝3.5Hz,2H),5.97(d,J＝6.5Hz,1H),4.24(s,2H),3.87(s,3H).Compound I-9a17 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8a17. 1 H NMR (400MHz, DMSO-d 6 ) δ11.56 (d, J = 6.7Hz, 1H), 11.14 (s, 1H), 10.85 (s, 1H), 8.22 (s, 1H), 7.76-7.68 ( m,2H),7.60(d,J＝7.8Hz,2H),7.55(d,J＝7.0Hz,1H),7.49(t,J＝7.5Hz,2H),7.34(d,J＝8.3Hz, 2H),7.09(d,J＝3.5Hz,2H),5.97(d,J＝6.5Hz,1H),4.24(s,2H),3.87(s,3H).
实施例18Example 18
2-(3-(5-氟-2-甲氧基苯基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a18)的制备2-(3-(5-fluoro-2-methoxyphenyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a18)
18.1中间体2-氯-N-((5-氟-2-甲氧基苯基)氨甲酰)乙酰胺(8a18)的制备18.1 Preparation of intermediate 2-chloro-N-((5-fluoro-2-methoxyphenyl)carbamoyl)acetamide (8a18)
按照实施例1中步骤1.2的方法合成中间体8a18。1H NMR(400MHz,DMSO-d6)δ11.14(s,1H),10.75(s,1H),8.00(d,J＝10.8Hz,1H),7.15-7.04(m,1H),6.90(t,J＝8.4Hz,1H),4.39(s,2H),3.93-3.86(m,3H).Intermediate 8a18 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.14 (s, 1H), 10.75 (s, 1H), 8.00 (d, J = 10.8Hz, 1H), 7.15-7.04 (m, 1H), 6.90 ( t,J＝8.4Hz,1H),4.39(s,2H),3.93-3.86(m,3H).
18.2终产物I-9a18的制备18.2 Preparation of final product I-9a18
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a18，制得化合物I-9a18。1H NMR(400MHz,DMSO-d6)δ11.57(d,J＝7.1Hz,1H),11.13(s,1H),10.87(s,1H),8.01(dd,J＝10.7,3.2Hz,1H),7.75-7.68(m,2H),7.61(d,J＝7.7Hz,2H),7.56(d,J＝7.7Hz,1H),7.49(t,J＝7.5Hz,2H),7.36-7.31(m,2H),7.06(dd,J＝9.2,5.2Hz,1H),6.88(td,J＝8.5,3.2Hz,1H),5.98(d,J＝7.0Hz,1H),4.24(d,J＝7.5Hz,2H),3.85(d,J＝7.7Hz,3H),3.40(s,3H).Compound I-9a18 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8a18. 1 H NMR (400MHz, DMSO-d 6 ) δ11.57(d,J＝7.1Hz,1H),11.13(s,1H),10.87(s,1H),8.01(dd,J＝10.7,3.2Hz, 1H),7.75-7.68(m,2H),7.61(d,J＝7.7Hz,2H),7.56(d,J＝7.7Hz,1H),7.49(t,J＝7.5Hz,2H),7.36- 7.31(m,2H),7.06(dd,J＝9.2,5.2Hz,1H),6.88(td,J＝8.5,3.2Hz,1H),5.98(d,J＝7.0Hz,1H),4.24(d ,J＝7.5Hz,2H),3.85(d,J＝7.7Hz,3H),3.40(s,3H).
实施例19Example 19
2-(3-(5-氯-2-硝基苯基)脲基)-2-氧代乙基-(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a19)的制备2-(3-(5-chloro-2-nitrophenyl)ureido)-2-oxoethyl-(1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a19)
19.1中间体2-氯-N-((5-氯-2-硝基苯基)氨甲酰)乙酰胺(8a19)的制备19.1 Preparation of intermediate 2-chloro-N-((5-chloro-2-nitrophenyl)carbamoyl)acetamide (8a19)
按照实施例1中步骤1.2的方法合成中间体8a19。1H NMR(400MHz,DMSO-d6)δ11.97(s,1H),11.36(s,1H),8.57(s,1H),8.21(d,1H),7.39(d,1H),4.41(s,2H)；13C NMR(150MHz,DMSO-d6)δ168.5,150.4,139.6,136.6,134.3,127.5,123.7,122.1,43.1；HR-MS(ESI):m/z[M-H]-calcd for C9H7Cl2N3O4:289.9741,found:289.9740.Intermediate 8a19 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.97(s,1H),11.36(s,1H),8.57(s,1H),8.21(d,1H),7.39(d,1H),4.41( s,2H);13C NMR(150MHz,DMSO-d6)δ168.5,150.4,139.6,136.6,134.3,127.5,123.7,122.1,43.1; HR-MS(ESI):m/z[MH]-calcd for C9H7Cl2N3O4: 289.9741,found:289.9740.
19.2终产物I-9a19的制备19.2 Preparation of final product I-9a19
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a19，制得化合物I-9a19。1H NMR(400MHz,DMSO-d6)δ12.09(s,1H),11.57(d,J＝6.5Hz,1H),11.33(s,1H),8.60(s,1H),8.19(d,J＝9.0Hz,1H),7.79-7.66(m,2H),7.64-7.44(m,5H),7.41-7.27(m,3H),5.97(d,J＝6.3Hz,1H),4.27(s,2H),3.39(s,3H)；13C NMR(150MHz,DMSO-d6)δ196.38,169.91,166.98,165.91,150.43,142.53,139.46,137.24,136.37,134.30,132.28,130.75,129.61,129.37,128.23,127.88,127.39,124.40,123.34,122.06,121.84,75.37,38.58,34.73；ESI-MS:m/z 596.6[M+H]+；C26H21ClN6O5S2,HRMS calcd.597.0776[M+H]+,found 597.0780. Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a19, and prepare compound I-9a19. 1 H NMR (400MHz, DMSO-d 6 ) δ12.09 (s, 1H), 11.57 (d, J = 6.5Hz, 1H), 11.33 (s, 1H), 8.60 (s, 1H), 8.19 (d, J＝9.0Hz,1H),7.79-7.66(m,2H),7.64-7.44(m,5H),7.41-7.27(m,3H),5.97(d,J＝6.3Hz,1H),4.27(s ,2H),3.39(s,3H); 13 C NMR (150MHz, DMSO-d 6 )δ196.38,169.91,166.98,165.91,150.43,142.53,139.46,137.24,136.37,134.30,132.28,130.75,129 .61,129.37, 128.23,127.88,127.39,124.40,123.34,122.06,121.84,75.37,38.58,34.73; ESI-MS: M/Z 596.6 [M + H] + ; C 26 H 21 ClN 6 O 5 S 2 , HRMS Calcd.597.0776666 [M+H] + ,found 597.0780.
实施例20Example 20
2-(3-(5-氯-2-硝基苯基)脲基)-2R-氧代乙基-(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代((R)-I-9a19)的制备2-(3-(5-chloro-2-nitrophenyl)ureido)-2R-oxoethyl-(1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio((R)-I-9a19)
按照实施例19中方法合成。鉴于化合物I-9a19结构中的七元氮环中含有一个手性C原子，它是具有一对对映体的混合物。我们对I-9a19进行了手性柱拆分并获得了光学纯化合物(R)-I-9a19，拆分方法如下：Synthesize according to the method in Example 19. Given the seven-membered nitrogen in the structure of compound I-9a19 The ring contains a chiral C atom, which is a mixture of a pair of enantiomers. We performed chiral column resolution on I-9a19 and obtained optically pure compound (R)-I-9a19. The resolution method is as follows:
Column：CHIRALPAK IG(IG00CE-UC011，Column size：0.46cm I.D.×25cm L，Column: CHIRALPAK IG (IG00CE-UC011, Column size: 0.46cm I.D.×25cm L,
Injection：2μL，Mobile phase：DCM/MeOH＝80/20(V/V)，Flow rate：1.0mL/min，Injection: 2μL, Mobile phase: DCM/MeOH=80/20(V/V), Flow rate: 1.0mL/min,
Wavelength：UV 254nm，Temperature：35℃，HPLC equipment：Shimadzu LC-20AT。Wavelength: UV 254nm, Temperature: 35℃, HPLC equipment: Shimadzu LC-20AT.
制得化合物(R)-I-9a19。1H NMR(400MHz,DMSO-d6)δ12.09(s,1H),11.57(d,J＝6.5Hz,1H),11.33(s,1H),8.60(s,1H),8.19(d,J＝9.0Hz,1H),7.79-7.66(m,2H),7.64-7.44(m,5H),7.41-7.27(m,3H),5.97(d,J＝6.3Hz,1H),4.27(s,2H),3.39(s,3H)；13C NMR(150MHz,DMSO-d6)δ196.38,169.91,166.98,165.91,150.43,142.53,139.46,137.24,136.37,134.30,132.28,130.75,129.61,129.37,128.23,127.88,127.39,124.40,123.34,122.06,121.84,75.37,38.58,34.73；ESI-MS:m/z 596.6[M+H]+；C26H21ClN6O5S2,HRMS calcd.597.0776[M+H]+,found 597.0780.l；HPLC column:t(R)＝4.187min,ee＝99.08％,[a]20 D＝+73.0(c＝0.1,CHCl3)Compound (R)-I-9a19 was prepared. 1 H NMR (400MHz, DMSO-d 6 ) δ12.09 (s, 1H), 11.57 (d, J = 6.5Hz, 1H), 11.33 (s, 1H), 8.60 (s, 1H), 8.19 (d, J＝9.0Hz,1H),7.79-7.66(m,2H),7.64-7.44(m,5H),7.41-7.27(m,3H),5.97(d,J＝6.3Hz,1H),4.27(s ,2H),3.39(s,3H); 13 C NMR (150MHz, DMSO-d 6 )δ196.38,169.91,166.98,165.91,150.43,142.53,139.46,137.24,136.37,134.30,132.28,130.75,129 .61,129.37, 128.23,127.88,127.39,124.40,123.34,122.06,121.84,75.37,38.58,34.73; ESI-MS: M/Z 596.6 [M + H] + ; C 26 H 21 ClN 6 O 5 S 2 , HRMS Calcd.597.0776666 [M+H] + , found 597.0780.l; HPLC column: t (R) = 4.187 min, ee = 99.08%, [a] 20 D = +73.0 (c = 0.1, CHCl 3 )
实施例21Example 21
2-(3-(5-氯-2-硝基苯基)脲基)-2S-氧代乙基-(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代((S)-I-9a19)的制备2-(3-(5-chloro-2-nitrophenyl)ureido)-2S-oxoethyl-(1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio((S)-I-9a19)
按照实施例20的步骤，制备与拆分制得化合物(S)-I-9a19。1H NMR(400MHz,DMSO-d6)δ12.09(s,1H),11.57(d,J＝6.5Hz,1H),11.33(s,1H),8.60(s,1H),8.19(d,J＝9.0Hz,1H),7.79-7.66(m,2H),7.64-7.44(m,5H),7.41-7.27(m,3H),5.97(d,J＝6.3Hz,1H),4.27(s,2H),3.39(s,3H)；13C NMR(150MHz,DMSO-d6)δ196.38,169.91,166.98,165.91,150.43,142.53,139.46,137.24,136.37,134.30,132.28,130.75,129.61,129.37,128.23,127.88,127.39,124.40,123.34,122.06,121.84,75.37,38.58,34.73；ESI-MS:m/z 596.6[M+H]+；C26H21ClN6O5S2,HRMS calcd.597.0776[M+H]+,found 597.0780；HPLC column:t(S)＝3.419min,ee＝99.68％,[a]20 D＝-68.0(c＝0.1,CHCl3).Compound (S)-I-9a19 was prepared according to the steps of Example 20, preparation and resolution. 1 H NMR (400MHz, DMSO-d 6 ) δ12.09 (s, 1H), 11.57 (d, J = 6.5Hz, 1H), 11.33 (s, 1H), 8.60 (s, 1H), 8.19 (d, J＝9.0Hz,1H),7.79-7.66(m,2H),7.64-7.44(m,5H),7.41-7.27(m,3H),5.97(d,J＝6.3Hz,1H),4.27(s ,2H),3.39(s,3H); 13 C NMR (150MHz, DMSO-d 6 )δ196.38,169.91,166.98,165.91,150.43,142.53,139.46,137.24,136.37,134.30,132.28,130.75,129 .61,129.37, 128.23,127.88,127.39,124.40,123.34,122.06,121.84,75.37,38.58,34.73; ESI-MS: M/Z 596.6 [M + H] + ; C 26 H 21 ClN 6 O 5 S 2 , HRMS Calcd.597.0776666 [M+H] + , found 597.0780; HPLC column: t(S)=3.419min, ee=99.68%, [a] 20 D =-68.0 (c=0.1, CHCl 3 ).
实施例22Example 22
2-(3-(2,4-二甲氧基苄基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9a20)的制备2-(3-(2,4-dimethoxybenzyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro- 1H-benzo[e][1,4]diazepine Preparation of -3-yl)carbodithio(I-9a20)
22.1中间体2-氯-N-((2,4-二甲氧基苄基)氨甲酰)乙酰胺(8a20)的制备22.1 Preparation of intermediate 2-chloro-N-((2,4-dimethoxybenzyl)carbamoyl)acetamide (8a20)
按照实施例1中步骤1.2的方法合成中间体8a20。1H NMR(400MHz,DMSO-d6)δ10.67(s,1H),10.00(s,1H),7.63-7.42(m,3H),4.88(s,2H),4.38(s,2H),3.79(s,3H),3.73(s,3H)；HR-MS(ESI):m/z[M-H]-calcd for C12H15ClN2O4:285.0648,found:285.0645.Intermediate 8a20 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.67(s,1H),10.00(s,1H),7.63-7.42(m,3H),4.88(s,2H),4.38(s,2H), 3.79(s,3H),3.73(s,3H); HR-MS(ESI):m/z[MH]-calcd for C12H15ClN2O4:285.0648,found:285.0645.
22.2终产物I-9a20的制备 22.2 Preparation of final product I-9a20
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8a20，制得化合物I-9a20。1H NMR(400MHz,DMSO-d6)δ11.48(s,1H),10.58(s,1H),8.47(s,1H),7.81-7.28(m,9H),7.06(d,J＝7.8Hz,1H),6.62-6.41(m,2H),5.98-5.95(m,1H),4.22(s,2H),4.14(s,2H),3.78(s,3H),3.72(s,3H),3.38(s,3H)；13C NMR(150MHz,DMSO-d6)δ196.52,169.36,166.93,165.92,159.87,157.82,152.49,142.53,137.24,132.28,130.74,129.59,129.34,129.08,128.23,127.87,124.40,122.07,118.33,104.26,98.27,75.24,55.33,55.04,38.46,37.90,34.72；ESI-MS:m/z 591.8[M+H]+；C29H29N5O5S2,HRMS calcd.592.1683[M+H]+,found 592.1685. Synthesize according to the method in step 1.3 in Example 1, replace the raw materials with the same equivalent of 8a20, and prepare compound I-9a20. 1 H NMR (400MHz, DMSO-d 6 ) δ11.48 (s, 1H), 10.58 (s, 1H), 8.47 (s, 1H), 7.81-7.28 (m, 9H), 7.06 (d, J = 7.8 Hz,1H),6.62-6.41(m,2H),5.98-5.95(m,1H),4.22(s,2H),4.14(s,2H),3.78(s,3H),3.72(s,3H) ,3.38(s,3H); 13 C NMR (150MHz, DMSO-d 6 )δ196.52,169.36,166.93,165.92,159.87,157.82,152.49,142.53,137.24,132.28,130.74,129.59,129.34, 129.08,128.23,127.87 ,124.40,122.07,118.33,104.26,98.27,75.24,55.33,55.04,38.46,37.90,34.72; ESI-MS:m/z 591.8[M+H] + ;C 29 H 29 N 5 O 5 S 2 ,HRMS calcd.592.1683[M+H] + ,found 592.1685 .
实施例23Example 23
2-氧代-2-(3-(吡啶-2-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b1)2-Oxo-2-(3-(pyridin-2-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[ e][1,4]diaza -3-yl)Carbodithio(I-9b1)
23.1中间体2-氯-N-(吡啶-2-基氨甲酰)乙酰胺(8b1)的制备23.1 Preparation of intermediate 2-chloro-N-(pyridin-2-ylcarbamoyl)acetamide (8b1)
按照实施例1中步骤1.2的方法合成中间体8b1。1H NMR(400MHz,DMSO-d6)δ11.21(s,1H),10.61(s,1H),8.31(s,1H),7.96-7.81(m,2H),7.15(d,J＝5.2Hz,1H),4.43(s,2H).23.2终产物I-9b1的制备Intermediate 8b1 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.21 (s, 1H), 10.61 (s, 1H), 8.31 (s, 1H), 7.96-7.81 (m, 2H), 7.15 (d, J = 5.2 Hz,1H),4.43(s,2H).23.2 Preparation of final product I-9b1
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b1，制得化合物I-9b1。1H NMR(400MHz,DMSO-d6)δ11.54(s,1H),11.14(s,1H),10.69(s,1H),8.28(s,2H),7.92(s,2H),7.79(s,2H),7.68(s,3H),7.51(m,8H),7.30(s,3H),7.11(s,1H),5.94(s,1H),4.24(s,3H),3.37(s,4H). Compound I-9b1 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8b1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.54(s,1H),11.14(s,1H),10.69(s,1H),8.28(s,2H),7.92(s,2H),7.79( s,2H),7.68(s,3H),7.51(m,8H),7.30(s,3H),7.11(s,1H),5.94(s,1H),4.24(s,3H),3.37(s ,4H) .
实施例24Example 24
2-(3-(5-氟吡啶-2-基)脲基)-2-氧乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b2)2-(3-(5-fluoropyridin-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzene And[e][1,4]diazapine -3-yl)Carbodithio(I-9b2)
24.1中间体2-氯-N-(5-氟吡啶-2-基)氨甲酰)乙酰胺(8b2)的制备24.1 Preparation of intermediate 2-chloro-N-(5-fluoropyridin-2-yl)carbamoyl)acetamide (8b2)
按照实施例1中步骤1.2的方法合成中间体8b2。1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),10.61(s,1H),8.33(d,J＝3.0Hz,1H),7.99(dd,J＝9.5,4.1Hz,1H),7.79(td,J＝8.6,2.9Hz,1H),4.41(s,2H).Intermediate 8b2 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.12 (s, 1H), 10.61 (s, 1H), 8.33 (d, J = 3.0Hz, 1H), 7.99 (dd, J = 9.5, 4.1Hz, 1H),7.79(td,J＝8.6,2.9Hz,1H),4.41(s,2H).
24.2终产物I-9b2的制备24.2 Preparation of final product I-9b2
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b2，制得化合物I-9b2。1H NMR(400MHz,DMSO-d6)δ11.51(s,1H),11.12(s,1H),10.74(s,1H),8.28(s,1H),7.97(s,1H),7.81-7.61(m,4H),7.60-7.40(m,5H),7.28(s,1H),5.92(s,1H),4.22(s,2H),3.35(s,3H).Compound I-9b2 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8b2. 1 H NMR (400MHz, DMSO-d 6 ) δ11.51(s,1H),11.12(s,1H),10.74(s,1H),8.28(s,1H),7.97(s,1H),7.81- 7.61(m,4H),7.60-7.40(m,5H),7.28(s,1H),5.92(s,1H),4.22(s,2H),3.35(s,3H).
实施例25Example 25
2-(3-(6-氯吡啶-2-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b3)2-(3-(6-chloropyridin-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H- Benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9b3)
25.1中间体2-氯-N-((6-氯吡啶-2-基)氨甲酰)乙酰胺(8b3)的制备25.1 Preparation of intermediate 2-chloro-N-((6-chloropyridin-2-yl)carbamoyl)acetamide (8b3)
按照实施例1中步骤1.2的方法合成中间体8b3。1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),10.60(s,1H),7.91(dd,J＝18.9,8.0Hz,2H),7.25(d,J＝7.5Hz,1H),4.42(s,2H). Intermediate 8b3 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.12 (s, 1H), 10.60 (s, 1H), 7.91 (dd, J = 18.9, 8.0Hz, 2H), 7.25 (d, J = 7.5Hz, 1H),4.42(s,2H).
23.2终产物I-9b3的制备23.2 Preparation of final product I-9b3
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b3，制得化合物I-9b3。1H NMR(400MHz,DMSO-d6)δ11.51(s,1H),11.15(s,1H),10.74(s,1H),7.90(d,J＝8.0Hz,1H),7.84(d,J＝7.9Hz,1H),7.66(s,2H),7.55(d,J＝7.5Hz,3H),7.45(d,J＝7.3Hz,2H),7.28(s,2H),7.20(d,J＝7.9Hz,1H),5.92(s,1H),4.23(s,2H),3.35(s,3H). Compound I-9b3 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8b3. 1 H NMR (400MHz, DMSO-d 6 ) δ11.51 (s, 1H), 11.15 (s, 1H), 10.74 (s, 1H), 7.90 (d, J = 8.0Hz, 1H), 7.84 (d, J＝7.9Hz,1H),7.66(s,2H),7.55(d,J＝7.5Hz,3H),7.45(d,J＝7.3Hz,2H),7.28(s,2H),7.20(d, J＝7.9Hz,1H),5.92(s,1H),4.23(s,2H),3.35(s,3H) .
实施例26Example 26
2-氧代-2-(3-(5-(三氟甲基)吡啶-2-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b4)2-Oxo-2-(3-(5-(trifluoromethyl)pyridin-2-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3- Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9b4)
26.1中间体2-氯-N-((5-(三氟甲基)吡啶-2-基)氨甲酰)乙酰胺(8b4)的制备26.1 Preparation of intermediate 2-chloro-N-((5-(trifluoromethyl)pyridin-2-yl)carbamoyl)acetamide (8b4)
按照实施例1中步骤1.2的方法合成中间体8b4。1H NMR(400MHz,DMSO-d6)δ11.25(s,1H),10.85(s,1H),8.74(s,1H),8.20(d,J＝31.3Hz,2H),4.46(s,2H).Intermediate 8b4 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.25 (s, 1H), 10.85 (s, 1H), 8.74 (s, 1H), 8.20 (d, J = 31.3Hz, 2H), 4.46 (s, 2H).
26.2终产物I-9b4的制备26.2 Preparation of final product I-9b4
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b4，制得化合物I-9b4。1H NMR(400MHz,DMSO-d6)δ11.53(s,1H),11.27(s,1H),10.98(s,1H),8.66(s,1H),8.19(d,J＝8.5Hz,1H),8.11(d,J＝6.7Hz,1H),7.70-7.64(m,2H),7.53(dd,J＝18.1,7.5Hz,3H),7.48-7.41(m,2H),7.28(t,J＝7.4Hz,2H),5.92(d,J＝8.2Hz,1H),4.24(s,2H),3.35(s,3H). Compound I-9b4 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8b4. 1 H NMR (400MHz, DMSO-d 6 ) δ11.53 (s, 1H), 11.27 (s, 1H), 10.98 (s, 1H), 8.66 (s, 1H), 8.19 (d, J = 8.5Hz, 1H),8.11(d,J＝6.7Hz,1H),7.70-7.64(m,2H),7.53(dd,J＝18.1,7.5Hz,3H),7.48-7.41(m,2H),7.28(t ,J＝7.4Hz,2H),5.92(d,J＝8.2Hz,1H),4.24(s,2H),3.35(s,3H) .
实施例27Example 27
2-(3-(3-氰基吡啶-2-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b5)2-(3-(3-cyanopyridin-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H -Benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9b5)
27.1中间体2-氯-N-((3-氰基吡啶-2-基)氨甲酰)乙酰胺(8b5)的制备27.1 Preparation of intermediate 2-chloro-N-((3-cyanopyridin-2-yl)carbamoyl)acetamide (8b5)
按照实施例1中步骤1.2的方法合成中间体8b5。1H NMR(400MHz,DMSO-d6)δ11.29(d,J＝8.0Hz,1H),10.61(s,1H),8.72(d,J＝5.3Hz,1H),8.40(d,J＝8.0Hz,1H),7.49(t,J＝6.5Hz,1H),4.46(d,J＝7.5Hz,2H).Intermediate 8b5 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.29 (d, J＝8.0Hz, 1H), 10.61 (s, 1H), 8.72 (d, J＝5.3Hz, 1H), 8.40 (d, J＝ 8.0Hz,1H),7.49(t,J＝6.5Hz,1H),4.46(d,J＝7.5Hz,2H).
27.2终产物I-9b5的制备27.2 Preparation of final product I-9b5
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b5，制得化合物I-9b5。1H NMR(400MHz,DMSO-d6)δ11.57(s,1H),11.30(d,J＝6.2Hz,1H),10.72(s,1H),8.69(s,1H),8.39(d,J＝8.2Hz,1H),7.74(dd,J＝13.3,6.9Hz,2H),7.61(t,J＝7.2Hz,2H),7.56(s,1H),7.50(q,J＝7.5,7.0Hz,3H),7.42-7.29(m,2H),5.99(t,J＝6.7Hz,1H),4.31(d,J＝6.5Hz,2H),3.41(d,J＝6.5Hz,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8b5, and prepare compound I-9b5. 1 H NMR (400MHz, DMSO-d 6 ) δ11.57 (s, 1H), 11.30 (d, J = 6.2Hz, 1H), 10.72 (s, 1H), 8.69 (s, 1H), 8.39 (d, J＝8.2Hz,1H),7.74(dd,J＝13.3,6.9Hz,2H),7.61(t,J＝7.2Hz,2H),7.56(s,1H),7.50(q,J＝7.5,7.0 Hz,3H),7.42-7.29(m,2H),5.99(t,J＝6.7Hz,1H),4.31(d,J＝6.5Hz,2H),3.41(d,J＝6.5Hz,3H) .
实施例28Example 28
2-(3-(6-氟吡啶-2-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b6)2-(3-(6-fluoropyridin-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H- Benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9b6)
28.1中间体2-氯-N-((6-氟吡啶-2-基)氨甲酰)乙酰胺(8b6)的制备28.1 Preparation of intermediate 2-chloro-N-((6-fluoropyridin-2-yl)carbamoyl)acetamide (8b6)
按照实施例1中步骤1.2的方法合成中间体8b6。1H NMR(400MHz,DMSO-d6)δ11.13(s,1H),10.54(s,1H),8.04-7.97(m,1H),7.88(dd,J＝8.0,2.3Hz,1H),6.92(dd,J＝8.0,2.3 Hz,1H),4.44(s,2H).Intermediate 8b6 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.13 (s, 1H), 10.54 (s, 1H), 8.04-7.97 (m, 1H), 7.88 (dd, J = 8.0, 2.3Hz, 1H), 6.92(dd,J＝8.0,2.3 Hz,1H),4.44(s,2H).
28.2终产物I-9b6的制备28.2 Preparation of final product I-9b6
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b6，制得化合物I-9b6。1H NMR(400MHz,DMSO-d6)δ11.55(d,J＝6.7Hz,1H),11.19(s,1H),10.71(s,1H),8.00(d,J＝8.3Hz,1H),7.89(d,J＝8.2Hz,1H),7.72(d,J＝10.7Hz,2H),7.60(d,J＝7.2Hz,2H),7.56(d,J＝6.3Hz,1H),7.50(d,J＝7.3Hz,2H),7.34(d,J＝6.9Hz,2H),6.90(d,J＝8.0Hz,1H),5.97(d,J＝6.5Hz,1H),4.28(s,2H),3.40(s,3H). Compound I-9b6 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8b6. 1 H NMR (400MHz, DMSO-d 6 ) δ11.55(d,J＝6.7Hz,1H),11.19(s,1H),10.71(s,1H),8.00(d,J＝8.3Hz,1H) ,7.89(d,J＝8.2Hz,1H),7.72(d,J＝10.7Hz,2H),7.60(d,J＝7.2Hz,2H),7.56(d,J＝6.3Hz,1H),7.50 (d,J＝7.3Hz,2H),7.34(d,J＝6.9Hz,2H),6.90(d,J＝8.0Hz,1H),5.97(d,J＝6.5Hz,1H),4.28(s ,2H),3.40(s,3H) .
实施例29Example 29
2-氧代-2-(3-(三氟甲基)吡啶-2-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b7)2-Oxo-2-(3-(trifluoromethyl)pyridin-2-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro- 1H-benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9b7)
29.1中间体2-氯-N-((3-(三氟甲基)吡啶-2-基)氨甲酰)乙酰胺(8b7)的制备29.1 Preparation of intermediate 2-chloro-N-((3-(trifluoromethyl)pyridin-2-yl)carbamoyl)acetamide (8b7)
按照实施例1中步骤1.2的方法合成中间体8b7。1H NMR(400MHz,DMSO-d6)δ11.26(s,1H),10.59(s,1H),8.75(s,1H),8.26(d,J＝7.3Hz,1H),7.53(s,1H),4.45(s,2H).Intermediate 8b7 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.26 (s, 1H), 10.59 (s, 1H), 8.75 (s, 1H), 8.26 (d, J = 7.3Hz, 1H), 7.53 (s, 1H),4.45(s,2H).
29.2终产物I-9b7的制备29.2 Preparation of final product I-9b7
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b7，制得化合物I-9b7。1H NMR(400MHz,DMSO-d6)δ11.55(s,1H),11.20(s,1H),10.72(s,1H),8.72(s,1H),8.24(d,J＝8.2Hz,1H),7.72(s,3H),7.60(s,2H),7.49(s,2H),7.34(s,2H),5.98(d,J＝6.5Hz,1H),4.28(s,2H),3.40(s,3H). Compound I-9b7 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8b7. 1 H NMR (400MHz, DMSO-d 6 ) δ11.55 (s, 1H), 11.20 (s, 1H), 10.72 (s, 1H), 8.72 (s, 1H), 8.24 (d, J = 8.2Hz, 1H),7.72(s,3H),7.60(s,2H),7.49(s,2H),7.34(s,2H),5.98(d,J＝6.5Hz,1H),4.28(s,2H), 3.40(s,3H) .
实施例30Example 30
2-氧代-2-(3-(6-(三氟甲基)吡啶-2-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b8)2-Oxo-2-(3-(6-(trifluoromethyl)pyridin-2-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3- Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9b8)
30.1中间体2-氯-N-((6-(三氟甲基)吡啶-2-基)氨甲酰)乙酰胺(8b8)的制备30.1 Preparation of intermediate 2-chloro-N-((6-(trifluoromethyl)pyridin-2-yl)carbamoyl)acetamide (8b8)
按照实施例1中步骤1.2的方法合成中间体8b8。1H NMR(400MHz,DMSO-d6)δ11.13(s,1H),10.76(d,J＝5.5Hz,1H),8.25(d,J＝7.8Hz,1H),8.12(t,J＝7.9Hz,1H),7.66(t,J＝6.3Hz,1H),4.46(d,J＝4.9Hz,2H).Intermediate 8b8 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.13 (s, 1H), 10.76 (d, J = 5.5Hz, 1H), 8.25 (d, J = 7.8Hz, 1H), 8.12 (t, J = 7.9Hz, 1H), 7.66 (t, J = 6.3Hz, 1H), 4.46 (d, J = 4.9Hz, 2H).
30.2终产物I-9b8的制备30.2 Preparation of final product I-9b8
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b8，制得化合物I-9b8。1H NMR(400MHz,DMSO-d6)δ11.57(s,1H),11.23(s,1H),10.93(d,J＝11.2Hz,1H),8.27(d,J＝8.7Hz,1H),8.12(s,1H),7.72(s,3H),7.64-7.59(m,3H),7.50(s,2H),7.34(s,2H),5.98(s,1H),4.30(s,2H),3.41(s,3H). Compound I-9b8 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8b8. 1 H NMR (400MHz, DMSO-d 6 ) δ11.57 (s, 1H), 11.23 (s, 1H), 10.93 (d, J = 11.2Hz, 1H), 8.27 (d, J = 8.7Hz, 1H) ,8.12(s,1H),7.72(s,3H),7.64-7.59(m,3H),7.50(s,2H),7.34(s,2H),5.98(s,1H),4.30(s,2H ),3.41(s,3H) .
实施例31Example 31
2-氧代-2-(3-(4-(三氟甲基)吡啶-2-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b9)2-Oxo-2-(3-(4-(trifluoromethyl)pyridin-2-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3- Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9b9)
31.1中间体2-氯-N-((4-(三氟甲基)吡啶-2-基)氨甲酰)乙酰胺(8b9)的制备31.1 Preparation of intermediate 2-chloro-N-((4-(trifluoromethyl)pyridin-2-yl)carbamoyl)acetamide (8b9)
按照实施例1中步骤1.2的方法合成中间体8b9。1H NMR(400MHz,DMSO-d6)δ11.24(s,1H),10.85(s,1H),8.63(d,J＝5.4Hz,1H),8.25(d,J＝5.4Hz,1H),7.55(d,J＝5.7Hz, 1H),4.46(d,J＝5.5Hz,2H).Intermediate 8b9 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.24 (s, 1H), 10.85 (s, 1H), 8.63 (d, J = 5.4Hz, 1H), 8.25 (d, J = 5.4Hz, 1H) ,7.55(d,J＝5.7Hz, 1H), 4.46 (d, J = 5.5Hz, 2H).
31.2终产物I-9b9的制备31.2 Preparation of final product I-9b9
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b9，制得化合物I-9b9。1H NMR(400MHz,DMSO-d6)δ11.52(s,1H),11.27(s,1H),11.00(s,1H),8.59(d,J＝4.8Hz,1H),8.26(s,1H),7.71(d,J＝10.9Hz,2H),7.60(d,J＝7.3Hz,2H),7.55(d,J＝6.4Hz,1H),7.49(d,J＝8.8Hz,3H),7.33(d,J＝7.2Hz,2H),5.97(d,J＝6.4Hz,1H),4.29(s,2H),3.39(s,3H). Compound I-9b9 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8b9. 1 H NMR (400MHz, DMSO-d 6 ) δ11.52 (s, 1H), 11.27 (s, 1H), 11.00 (s, 1H), 8.59 (d, J = 4.8Hz, 1H), 8.26 (s, 1H),7.71(d,J＝10.9Hz,2H),7.60(d,J＝7.3Hz,2H),7.55(d,J＝6.4Hz,1H),7.49(d,J＝8.8Hz,3H) ,7.33(d,J＝7.2Hz,2H),5.97(d,J＝6.4Hz,1H),4.29(s,2H),3.39(s,3H) .
实施例32Example 32
2-(3-(3-氟吡啶-2-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b10)2-(3-(3-fluoropyridin-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H- Benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9b10)
32.1中间体2-氯-N-((3-氟吡啶-2-基)氨甲酰)乙酰胺(8b10)的制备32.1 Preparation of intermediate 2-chloro-N-((3-fluoropyridin-2-yl)carbamoyl)acetamide (8b10)
按照实施例1中步骤1.2的方法合成中间体8b10。1H NMR(400MHz,DMSO-d6)δ11.26(s,1H),10.23(s,1H),8.26(d,J＝4.7Hz,1H),7.82(t,J＝9.3Hz,1H),7.36(dt,J＝8.6,4.4Hz,1H),4.45(s,2H).Intermediate 8b10 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.26 (s, 1H), 10.23 (s, 1H), 8.26 (d, J = 4.7Hz, 1H), 7.82 (t, J = 9.3Hz, 1H) ,7.36(dt,J＝8.6,4.4Hz,1H),4.45(s,2H).
32.2终产物I-9b10的制备32.2 Preparation of final product I-9b10
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b10，制得化合物I-9b10。1H NMR(400MHz,DMSO-d6)δ11.55(s,1H),11.22(s,1H),10.35(s,1H),8.25(d,J＝4.3Hz,1H),7.84-7.69(m,4H),7.63-7.58(m,2H),7.56(d,J＝3.4Hz,1H),7.50(dd,J＝7.5,4.1Hz,2H),7.37-7.33(m,2H),5.99(dt,J＝6.7,3.7Hz,1H),4.36-4.25(m,2H),3.40(s,3H).Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8b10, and prepare compound I-9b10. 1 H NMR (400MHz, DMSO-d 6 ) δ11.55 (s, 1H), 11.22 (s, 1H), 10.35 (s, 1H), 8.25 (d, J = 4.3Hz, 1H), 7.84-7.69 ( m,4H),7.63-7.58(m,2H),7.56(d,J＝3.4Hz,1H),7.50(dd,J＝7.5,4.1Hz,2H),7.37-7.33(m,2H),5.99 (dt,J＝6.7,3.7Hz,1H),4.36-4.25(m,2H),3.40(s,3H).
实施例33Example 33
2-(3-(3-硝基吡啶-2-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b11)2-(3-(3-nitropyridin-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H -Benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9b11)
33.1中间体2-氯-N-((3-硝基吡啶-2-基)氨甲酰)乙酰胺(8b11)的制备33.1 Preparation of intermediate 2-chloro-N-((3-nitropyridin-2-yl)carbamoyl)acetamide (8b11)
按照实施例1中步骤1.2的方法合成中间体8b11。1H NMR(400MHz,DMSO-d6)δ11.27(d,J＝7.3Hz,1H),11.08(d,J＝6.8Hz,1H),8.73(d,J＝5.1Hz,1H),8.51(d,J＝8.1Hz,1H),7.51(dd,J＝8.3,4.7Hz,1H),4.44(d,J＝8.0Hz,2H).Intermediate 8b11 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.27(d,J＝7.3Hz,1H),11.08(d,J＝6.8Hz,1H),8.73(d,J＝5.1Hz,1H),8.51 (d, J＝8.1Hz, 1H), 7.51 (dd, J＝8.3, 4.7Hz, 1H), 4.44 (d, J＝8.0Hz, 2H).
33.2终产物I-9b11的制备33.2 Preparation of final product I-9b11
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b11，制得化合物I-9b11。1H NMR(400MHz,DMSO-d6)δ11.59(s,1H),11.31(s,1H),11.20(s,1H),8.70(s,1H),8.49(s,1H),7.72(s,2H),7.60(s,2H),7.57(s,1H),7.52–7.48(m,3H),7.34(s,2H),5.99(d,J＝5.6Hz,1H),4.30(s,2H),3.41(d,J＝6.4Hz,3H).Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8b11, and prepare compound I-9b11. 1 H NMR (400MHz, DMSO-d 6 ) δ11.59(s,1H),11.31(s,1H),11.20(s,1H),8.70(s,1H),8.49(s,1H),7.72( s,2H),7.60(s,2H),7.57(s,1H),7.52–7.48(m,3H),7.34(s,2H),5.99(d,J＝5.6Hz,1H),4.30(s ,2H),3.41(d,J＝6.4Hz,3H).
实施例34Example 34
2-(3-(6-氯-3-硝基吡啶-2-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9b12)2-(3-(6-chloro-3-nitropyridin-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3- Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9b12)
34.1中间体2-氯-N-((6-氯-3-硝基吡啶-2-基)氨甲酰)乙酰胺(8b12)的制备34.1 Preparation of intermediate 2-chloro-N-((6-chloro-3-nitropyridin-2-yl)carbamoyl)acetamide (8b12)
按照实施例1中步骤1.2的方法合成中间体8b12。1H NMR(400MHz,DMSO-d6)δ 11.46(s,1H),11.28(s,1H),8.56(dd,J＝8.5,1.4Hz,1H),7.59-7.52(m,1H),4.43(d,J＝1.4Hz,2H).Intermediate 8b12 was synthesized according to the method of step 1.2 in Example 1. 1H NMR (400MHz, DMSO-d 6 )δ 11.46(s,1H),11.28(s,1H),8.56(dd,J＝8.5,1.4Hz,1H),7.59-7.52(m,1H),4.43(d,J＝1.4Hz,2H).
34.2终产物I-9b12的制备34.2 Preparation of final product I-9b12
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8b12，制得化合物I-9b12。1H NMR(400MHz,DMSO-d6)δ11.54(d,J＝20.4Hz,1H),11.29(s,1H),9.81(s,1H),8.53(d,J＝7.9Hz,1H),8.45(d,J＝10.9Hz,1H),7.70(s,2H),7.59(d,J＝7.5Hz,1H),7.53(d,J＝11.7Hz,2H),7.48(d,J＝8.1Hz,2H),7.33(d,J＝7.7Hz,2H),5.97(d,J＝6.3Hz,1H),4.28(s,2H),3.39(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8b12, and prepare compound I-9b12. 1 H NMR (400MHz, DMSO-d 6 ) δ11.54(d,J＝20.4Hz,1H),11.29(s,1H),9.81(s,1H),8.53(d,J＝7.9Hz,1H) ,8.45(d,J＝10.9Hz,1H),7.70(s,2H),7.59(d,J＝7.5Hz,1H),7.53(d,J＝11.7Hz,2H),7.48(d,J＝ 8.1Hz,2H),7.33(d,J＝7.7Hz,2H),5.97(d,J＝6.3Hz,1H),4.28(s,2H),3.39(s,3H) .
实施例35Example 35
2-氧代-2-(3-(吡啶-3-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9c1)2-Oxo-2-(3-(pyridin-3-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[ e][1,4]diaza -3-yl)Carbodithio(I-9c1)
35.1中间体2-氯-N-(吡啶-3-基氨甲酰)乙酰胺(8c1)的制备35.1 Preparation of intermediate 2-chloro-N-(pyridin-3-ylcarbamoyl)acetamide (8c1)
按照实施例1中步骤1.2的方法合成中间体8c1。1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),10.19(s,1H),8.70(d,J＝2.6Hz,1H),8.30(dd,J＝4.7,1.5Hz,1H),8.00(ddd,J＝8.1,2.6,1.4Hz,1H),7.36(dd,J＝8.4,4.7Hz,1H),4.40(s,2H).Intermediate 8c1 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.01 (s, 1H), 10.19 (s, 1H), 8.70 (d, J = 2.6Hz, 1H), 8.30 (dd, J = 4.7, 1.5Hz, 1H), 8.00 (ddd, J＝8.1, 2.6, 1.4Hz, 1H), 7.36 (dd, J＝8.4, 4.7Hz, 1H), 4.40 (s, 2H).
35.2终产物I-9c1的制备35.2 Preparation of final product I-9c1
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8c1，制得化合物I-9c1。1H NMR(400MHz,DMSO-d6)δ11.52(d,J＝5.7Hz,1H),11.05(s,1H),10.34(s,1H),8.68(d,J＝8.9Hz,1H),8.28(d,J＝3.4Hz,1H),7.98(d,J＝4.8Hz,1H),7.76-7.67(m,2H),7.59(d,J＝6.7Hz,2H),7.54(d,J＝6.8Hz,1H),7.52-7.45(m,2H),7.37-7.30(m,3H),5.97(d,J＝6.3Hz,1H),4.26(s,2H),3.39(s,3H). Synthesize according to the method in step 1.3 in Example 1, replace the raw materials with the same equivalent of 8c1, and prepare compound I-9c1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.52(d,J＝5.7Hz,1H),11.05(s,1H),10.34(s,1H),8.68(d,J＝8.9Hz,1H) ,8.28(d,J＝3.4Hz,1H),7.98(d,J＝4.8Hz,1H),7.76-7.67(m,2H),7.59(d,J＝6.7Hz,2H),7.54(d, J＝6.8Hz,1H),7.52-7.45(m,2H),7.37-7.30(m,3H),5.97(d,J＝6.3Hz,1H),4.26(s,2H),3.39(s,3H ) .
实施例36Example 36
2-(3-(2-甲氧基吡啶-3-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9c2)2-(3-(2-methoxypyridin-3-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro- 1H-benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9c2)
36.1中间体2-氯-N-((2-甲氧基吡啶-3-基)氨甲酰)乙酰胺(8c2)的制备36.1 Preparation of intermediate 2-chloro-N-((2-methoxypyridin-3-yl)carbamoyl)acetamide (8c2)
按照实施例1中步骤1.2的方法合成中间体8c2。1H NMR(400MHz,DMSO-d6)δ11.16(s,1H),10.55(s,1H),8.38(d,J＝7.7Hz,1H),7.86(d,J＝4.8Hz,1H),7.04–6.96(m,1H),4.38(s,2H),3.95(s,3H).Intermediate 8c2 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.16 (s, 1H), 10.55 (s, 1H), 8.38 (d, J = 7.7Hz, 1H), 7.86 (d, J = 4.8Hz, 1H) ,7.04–6.96(m,1H),4.38(s,2H),3.95(s,3H).
36.2终产物I-9c2的制备36.2 Preparation of final product I-9c2
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8c2，制得化合物I-9c2。1H NMR(400MHz,DMSO-d6)δ11.53(d,J＝8.0Hz,1H),11.11(s,1H),10.68(s,1H),8.39(d,J＝7.7Hz,1H),7.83(d,J＝6.8Hz,1H),7.74-7.66(m,2H),7.59(d,J＝8.6Hz,2H),7.54(d,J＝7.9Hz,1H),7.47(t,J＝8.1Hz,2H),7.34-7.28(m,2H),7.02-6.91(m,1H),6.01-5.93(m,1H),4.23(s,2H),3.93(s,3H),3.38(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8c2, and prepare compound I-9c2. 1 H NMR (400MHz, DMSO-d 6 ) δ11.53(d,J＝8.0Hz,1H),11.11(s,1H),10.68(s,1H),8.39(d,J＝7.7Hz,1H) ,7.83(d,J＝6.8Hz,1H),7.74-7.66(m,2H),7.59(d,J＝8.6Hz,2H),7.54(d,J＝7.9Hz,1H),7.47(t, J＝8.1Hz,2H),7.34-7.28(m,2H),7.02-6.91(m,1H),6.01-5.93(m,1H),4.23(s,2H),3.93(s,3H),3.38 (s,3H) .
实施例37Example 37
2-氧代-2-(3-(4-(三氟甲基)吡啶-3-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯 并[e][1,4]二氮杂-3-基)碳二硫代(I-9c3)2-Oxo-2-(3-(4-(trifluoromethyl)pyridin-3-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3- Dihydro-1H-benzene And[e][1,4]diazapine -3-yl)Carbodithio(I-9c3)
37.1中间体2-氯-N-((4-(三氟甲基)吡啶-3-基)氨甲酰)乙酰胺(8c3)的制备37.1 Preparation of intermediate 2-chloro-N-((4-(trifluoromethyl)pyridin-3-yl)carbamoyl)acetamide (8c3)
按照实施例1中步骤1.2的方法合成中间体8c3。1H NMR(400MHz,DMSO-d6)δ11.42(s,1H),10.63(s,1H),9.28(s,1H),8.64(d,J＝5.1Hz,1H),7.80(d,J＝5.0Hz,1H),4.44(s,2H).Intermediate 8c3 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.42 (s, 1H), 10.63 (s, 1H), 9.28 (s, 1H), 8.64 (d, J = 5.1Hz, 1H), 7.80 (d, J＝5.0Hz,1H),4.44(s,2H).
37.2终产物I-9c3的制备37.2 Preparation of final product I-9c3
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8c3，制得化合物I-9c3。1H NMR(400MHz,DMSO-d6)δ11.59(d,J＝6.5Hz,1H),11.39(s,1H),10.79(s,1H),9.31(d,J＝8.7Hz,1H),8.67-8.59(m,1H),7.8-7.72(m,3H),7.62(d,J＝7.4Hz,2H),7.58(s,1H),7.51(t,J＝7.3Hz,2H),7.36(d,J＝9.0Hz,2H),5.99(d,J＝6.7Hz,1H),4.32(s,2H),3.42(s,3H).Synthesize according to the method in step 1.3 in Example 1, replace the raw materials with the same equivalent of 8c3, and prepare compound I-9c3. 1 H NMR (400MHz, DMSO-d 6 ) δ11.59(d,J＝6.5Hz,1H),11.39(s,1H),10.79(s,1H),9.31(d,J＝8.7Hz,1H) ,8.67-8.59(m,1H),7.8-7.72(m,3H),7.62(d,J＝7.4Hz,2H),7.58(s,1H),7.51(t,J＝7.3Hz,2H), 7.36(d,J＝9.0Hz,2H),5.99(d,J＝6.7Hz,1H),4.32(s,2H),3.42(s,3H).
实施例38Example 38
2-(3-((2-甲氧基吡啶-3-基)甲基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9c4)2-(3-((2-methoxypyridin-3-yl)methyl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3 -Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9c4)
38.1中间体2-氯-N-(((2-甲氧基吡啶-3-基)甲基)氨甲酰)乙酰胺(8c4)的制备38.1 Preparation of intermediate 2-chloro-N-(((2-methoxypyridin-3-yl)methyl)carbamoyl)acetamide (8c4)
按照实施例1中步骤1.2的方法合成中间体8c4。1H NMR(400MHz,DMSO-d6)δ10.69(s,1H),8.55(s,1H),8.12–8.02(m,1H),7.53(d,J＝7.3Hz,1H),6.97(dd,J＝7.2,5.1Hz,1H),4.30(d,J＝7.6Hz,4H),3.90(s,3H).Intermediate 8c4 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.69 (s, 1H), 8.55 (s, 1H), 8.12–8.02 (m, 1H), 7.53 (d, J = 7.3Hz, 1H), 6.97 ( dd,J＝7.2,5.1Hz,1H),4.30(d,J＝7.6Hz,4H),3.90(s,3H).
38.2终产物I-9c4的制备38.2 Preparation of final product I-9c4
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8c4，制得化合物I-9c4。ESI-MS:m/z 563.2[M+H]+.Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8c4, and prepare compound I-9c4. ESI-MS:m/z 563.2[M+H] + .
实施例39Example 39
2-(3-(6-氯嘧啶-4-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9d1)2-(3-(6-chloropyrimidin-4-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H- Benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9d1)
39.1中间体2-氯-N-((6-氯嘧啶-4-基)氨甲酰)乙酰胺(8d1)的制备39.1 Preparation of intermediate 2-chloro-N-((6-chloropyrimidin-4-yl)carbamoyl)acetamide (8d1)
按照实施例1中步骤1.2的方法合成中间体8d1。1H NMR(400MHz,DMSO-d6)δ11.30(s,1H),10.85(s,1H),8.77(s,1H),7.97(d,J＝2.7Hz,1H),4.46(d,J＝2.4Hz,2H).Intermediate 8d1 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.30 (s, 1H), 10.85 (s, 1H), 8.77 (s, 1H), 7.97 (d, J = 2.7Hz, 1H), 4.46 (d, J＝2.4Hz,2H).
39.2终产物I-9d1的制备39.2 Preparation of final product I-9d1
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8d1，制得化合物I-9d1。1H NMR(400MHz,DMSO-d6)δ11.52(s,1H),11.38(s,1H),10.99(s,1H),8.71(s,1H),7.94(s,1H),7.70-7.63(m,2H),7.57-7.50(m,3H),7.45(d,J＝8.0Hz,2H),7.28(s,2H),5.91(s,1H),4.24(s,2H),3.34(d,J＝3.2Hz,3H). Compound I-9d1 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8d1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.52(s,1H),11.38(s,1H),10.99(s,1H),8.71(s,1H),7.94(s,1H),7.70- 7.63(m,2H),7.57-7.50(m,3H),7.45(d,J＝8.0Hz,2H),7.28(s,2H),5.91(s,1H),4.24(s,2H),3.34 (d,J=3.2Hz,3H) .
实施例40Example 40
2-氧代-2-(3-(2-(三氟甲基)嘧啶-5-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9e1)2-Oxo-2-(3-(2-(trifluoromethyl)pyrimidin-5-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3- Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9e1)
40.1中间体2-氯-N-((2-(三氟甲基)嘧啶-5-基)氨甲酰)乙酰胺(8e1)的制备 40.1 Preparation of intermediate 2-chloro-N-((2-(trifluoromethyl)pyrimidin-5-yl)carbamoyl)acetamide (8e1)
按照实施例1中步骤1.2的方法合成中间体8e1。1H NMR(400MHz,DMSO-d6)δ11.26(s,1H),10.55(s,1H),9.26(s,2H),4.45(s,2H).Intermediate 8e1 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.26(s,1H),10.55(s,1H),9.26(s,2H),4.45(s,2H).
40.2终产物I-9e1的制备40.2 Preparation of final product I-9e1
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8e1，制得化合物I-9e1。1H NMR(400MHz,DMSO-d6)δ11.53(s,1H),11.28(s,1H),10.66(s,1H),9.21(s,2H),7.68(s,2H),7.57(s,2H),7.45(s,2H),7.29(s,2H),5.93(s,1H),4.26(s,2H),3.35(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8e1, and prepare compound I-9e1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.53(s,1H),11.28(s,1H),10.66(s,1H),9.21(s,2H),7.68(s,2H),7.57( s,2H),7.45(s,2H),7.29(s,2H),5.93(s,1H),4.26(s,2H),3.35(s,3H) .
实施例41Example 41
2-氧代-2-(3-(吡嗪-2-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9f1)2-Oxo-2-(3-(pyrazin-2-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo [e][1,4]diazapine -3-yl)Carbodithio (I-9f1)
41.1中间体2-氯-N-(吡嗪-2-基氨甲酰)乙酰胺(8f1)的制备41.1 Preparation of intermediate 2-chloro-N-(pyrazin-2-ylcarbamoyl)acetamide (8f1)
按照实施例1中步骤1.2的方法合成中间体8f1。1H NMR(400MHz,DMSO-d6)δ11.21(s,1H),10.62(s,1H),9.21(s,1H),8.40(d,J＝2.1Hz,2H),4.43(d,J＝2.1Hz,2H).Intermediate 8f1 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.21 (s, 1H), 10.62 (s, 1H), 9.21 (s, 1H), 8.40 (d, J = 2.1Hz, 2H), 4.43 (d, J＝2.1Hz,2H).
41.2终产物I-9f1的制备41.2 Preparation of final product I-9f1
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8f1，制得化合物I-9f1。1H NMR(400MHz,DMSO-d6)δ10.78(s,1H),9.22(s,1H),8.38(d,J＝10.8Hz,2H),7.73-7.65(m,2H),7.57(d,J＝6.0Hz,2H),7.52(d,J＝6.4Hz,1H),7.48-7.43(m,3H),7.30(q,J＝8.2Hz,3H),5.93(s,1H),4.25(s,2H),3.36(s,3H). Compound I-9f1 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8f1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.78 (s, 1H), 9.22 (s, 1H), 8.38 (d, J = 10.8Hz, 2H), 7.73-7.65 (m, 2H), 7.57 ( d,J＝6.0Hz,2H),7.52(d,J＝6.4Hz,1H),7.48-7.43(m,3H),7.30(q,J＝8.2Hz,3H),5.93(s,1H), 4.25(s,2H),3.36(s,3H) .
实施例42Example 42
2-(3-(1H-吡唑基-5-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9g1)2-(3-(1H-pyrazolyl-5-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H -Benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9g1)
42.1中间体N-((1H-吡唑基-5-基)氨甲酰)-2-氯乙酰胺(8g1)的制备42.1 Preparation of intermediate N-((1H-pyrazolyl-5-yl)carbamoyl)-2-chloroacetamide (8g1)
按照实施例1中步骤1.2的方法合成中间体8g1。1H NMR(400MHz,DMSO-d6)δ12.42(s,1H),10.89(s,1H),10.27(s,1H),7.657.54(m,1H),6.42-6.29(m,1H),4.35(d,J＝7.8Hz,2H).Intermediate 8g1 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ12.42(s,1H),10.89(s,1H),10.27(s,1H),7.657.54(m,1H),6.42-6.29(m,1H ),4.35(d,J＝7.8Hz,2H).
42.2终产物I-9g1的制备42.2 Preparation of final product I-9g1
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8g1，制得化合物I-9g1。1H NMR(400MHz,DMSO-d6)δ11.52(s,1H),11.14(s,1H),10.65(s,1H),8.16(s,1H),7.71(d,J＝8.0Hz,3H),7.59-7.51(m,3H),7.48(d,J＝5.2Hz,2H),7.30(s,2H),6.67(s,1H),5.95(s,1H),4.21(s,2H),3.37(s,3H). Compound I-9g1 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8g1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.52 (s, 1H), 11.14 (s, 1H), 10.65 (s, 1H), 8.16 (s, 1H), 7.71 (d, J = 8.0Hz, 3H),7.59-7.51(m,3H),7.48(d,J＝5.2Hz,2H),7.30(s,2H),6.67(s,1H),5.95(s,1H),4.21(s,2H ),3.37(s,3H) .
实施例43Example 43
2-氧代-2-(3-(噻唑-2-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9g2)2-Oxo-2-(3-(thiazol-2-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[ e][1,4]diaza -3-yl)Carbodithio(I-9g2)
43.1中间体2-氯-N-(噻唑-2-基氨甲酰)乙酰胺(8g2)的制备43.1 Preparation of intermediate 2-chloro-N-(thiazol-2-ylcarbamoyl)acetamide (8g2)
按照实施例1中步骤1.2的方法合成中间体8g2。ESI-MS:m/z 219.9[M+H]+ Intermediate 8g2 was synthesized according to the method of step 1.2 in Example 1. ESI-MS:m/z 219.9[M+H] +
43.2终产物I-9g2的制备 43.2 Preparation of final product I-9g2
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8g2，制得化合物I-9g2。1H NMR(400MHz,DMSO-d6)δ11.53(d,J＝6.9Hz,1H),11.40(s,2H),7.69(d,J＝10.1Hz,2H),7.56(dd,J＝13.5,7.2Hz,3H),7.46(dq,J＝12.6,5.8,3.4Hz,3H),7.29(t,J＝13.3Hz,3H),6.00–5.88(m,1H),4.28(d,J＝9.4Hz,2H),3.36(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8g2, and prepare compound I-9g2. 1 H NMR (400MHz, DMSO-d 6 ) δ11.53 (d, J = 6.9 Hz, 1H), 11.40 (s, 2H), 7.69 (d, J = 10.1 Hz, 2H), 7.56 (dd, J = 13.5,7.2Hz,3H),7.46(dq,J＝12.6,5.8,3.4Hz,3H),7.29(t,J＝13.3Hz,3H),6.00–5.88(m,1H),4.28(d,J ＝9.4Hz,2H),3.36(s,3H) .
实施例44Example 44
2-(3-甲基-1,2,4-噻二唑-5-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9g3)2-(3-methyl-1,2,4-thiadiazol-5-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2, 3-Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9g3)
44.1中间体2-氯-N-((3-甲基-1,2,4-噻二唑-5-基)氨甲酰)乙酰胺(8g3)的制备44.1 Preparation of intermediate 2-chloro-N-((3-methyl-1,2,4-thiadiazol-5-yl)carbamoyl)acetamide (8g3)
按照实施例1中步骤1.2的方法合成中间体8g3。1H NMR(400MHz,DMSO-d6)δ11.62(s,1H),11.45(s,1H),4.52(s,2H),2.45(s,3H).Intermediate 8g3 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.62(s,1H),11.45(s,1H),4.52(s,2H),2.45(s,3H).
44.2终产物I-9g3的制备44.2 Preparation of final product I-9g3
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8g3，制得化合物I-9g3。1H NMR(400MHz,DMSO-d6)δ11.65(s,1H),11.59(s,1H),11.53(d,J＝3.9Hz,1H),7.71(d,J＝7.6Hz,2H),7.58(d,J＝8.2Hz,3H),7.48(d,J＝7.3Hz,3H),7.32(s,2H),5.95(d,J＝5.5Hz,1H),4.32(s,2H),3.38(s,3H),2.42(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8g3, and prepare compound I-9g3. 1 H NMR (400MHz, DMSO-d 6 ) δ11.65 (s, 1H), 11.59 (s, 1H), 11.53 (d, J = 3.9Hz, 1H), 7.71 (d, J = 7.6Hz, 2H) ,7.58(d,J＝8.2Hz,3H),7.48(d,J＝7.3Hz,3H),7.32(s,2H),5.95(d,J＝5.5Hz,1H),4.32(s,2H) ,3.38(s,3H),2.42(s,3H) .
实施例45Example 45
2-(3-(噁唑-2-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9g4)2-(3-(oxazol-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo [e][1,4]diazapine -3-yl)Carbodithio(I-9g4)
45.1中间体2-氯-N-(噁唑-2-基氨甲酰)乙酰胺(8g4)的制备45.1 Preparation of intermediate 2-chloro-N-(oxazol-2-ylcarbamoyl)acetamide (8g4)
按照实施例1中步骤1.2的方法合成中间体8g4。1H NMR(400MHz,DMSO-d6)δ11.22(s,1H),10.93(s,1H),7.92(s,1H),7.15(s,1H),4.52(s,2H).Intermediate 8g4 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.22(s,1H),10.93(s,1H),7.92(s,1H),7.15(s,1H),4.52(s,2H).
45.2终产物I-9g4的制备45.2 Preparation of final product I-9g4
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8g4，制得化合物I-9g4。ESI-MS:m/z 509.1[M+H]+.Compound I-9g4 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8g4. ESI-MS:m/z 509.1[M+H] + .
实施例46Example 46
2-(3-(5-甲基-1,3,4-噁二唑-2-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9g5)2-(3-(5-methyl-1,3,4-oxadiazol-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl -2,3-dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9g5)
46.1中间体2-氯-N-((5-甲基-1,3,4-噁二唑-2-基)氨甲酰)乙酰胺(8g5)的制备46.1 Preparation of intermediate 2-chloro-N-((5-methyl-1,3,4-oxadiazol-2-yl)carbamoyl)acetamide (8g5)
按照实施例1中步骤1.2的方法合成中间体8g5。1H NMR(400MHz,DMSO-d6)δ11.17(s,1H),10.87(s,1H),4.47(s,2H),3.34(s,3H).Intermediate 8g5 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.17(s,1H),10.87(s,1H),4.47(s,2H),3.34(s,3H).
46.2终产物I-9g5的制备46.2 Preparation of final product I-9g5
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8g5，制得化合物I-9g5。1H NMR(400MHz,DMSO-d6)δ11.52(d,J＝7.8Hz,1H),11.27(s,1H),10.90(s,1H),7.76-7.67(m,2H),7.58(d,J＝7.6Hz,2H),7.53(d,J＝6.9Hz,1H),7.47(t,J＝7.3Hz,2H),7.32(q,J＝8.0,7.1Hz,2H),6.02–5.91(m,1H),4.28(s,2H),3.38(s,3H),2.44(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8g5, and prepare compound I-9g5. 1 H NMR (400MHz, DMSO-d 6 ) δ11.52 (d, J = 7.8 Hz, 1H), 11.27 (s, 1H), 10.90 (s, 1H), 7.76-7.67 (m, 2H), 7.58 ( d,J＝7.6Hz,2H),7.53(d,J＝6.9Hz,1H),7.47(t,J＝7.3Hz,2H),7.32(q,J＝8.0,7.1Hz,2H),6.02– 5.91(m,1H),4.28(s,2H),3.38(s,3H),2.44(s,3H) .
实施例47Example 47
2-(3-(4-甲基噁唑基-2-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9g6)2-(3-(4-methyloxazol-2-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-benzo[e][1,4]diazepine -3-yl)Carbodithio (I-9g6)
47.1中间体2-氯-N-((4-甲基噁唑-2-基)氨甲酰)乙酰胺(8g6)的制备47.1 Preparation of intermediate 2-chloro-N-((4-methyloxazol-2-yl)carbamoyl)acetamide (8g6)
按照实施例1中步骤1.2的方法合成中间体8g6。1H NMR(400MHz,DMSO-d6)δ11.28(s,1H),10.95(s,1H),7.43(s,1H),4.56(d,J＝9.3Hz,2H),2.07(d,J＝9.7Hz,3H).Intermediate 8g6 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.28 (s, 1H), 10.95 (s, 1H), 7.43 (s, 1H), 4.56 (d, J = 9.3Hz, 2H), 2.07 (d, J＝9.7Hz,3H).
47.2终产物I-9g6的制备47.2 Preparation of final product I-9g6
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8g6，制得化合物I-9g6。1H NMR(400MHz,DMSO-d6)δ11.52(s,1H),11.26(s,1H),10.88(s,1H),7.72(d,J＝9.6Hz,2H),7.62-7.57(m,3H),7.55(s,1H),7.49(q,J＝7.9,5.9Hz,2H),7.34(d,J＝7.4Hz,2H),5.96(d,J＝6.7Hz,1H),4.32(s,2H),3.39(s,3H),2.04(d,J＝3.6Hz,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8g6, and prepare compound I-9g6. 1 H NMR (400MHz, DMSO-d 6 ) δ11.52 (s, 1H), 11.26 (s, 1H), 10.88 (s, 1H), 7.72 (d, J = 9.6Hz, 2H), 7.62-7.57 ( m,3H),7.55(s,1H),7.49(q,J＝7.9,5.9Hz,2H),7.34(d,J＝7.4Hz,2H),5.96(d,J＝6.7Hz,1H), 4.32(s,2H),3.39(s,3H),2.04(d,J=3.6Hz,3H) .
实施例48Example 48
2-氧代-2-(3-(4-(三氟甲基)噁唑-2-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9g7)2-Oxo-2-(3-(4-(trifluoromethyl)oxazol-2-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3 -Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9g7)
48.1中间体2-氯-N-((4-(三氟甲基)噁唑-2-基)氨甲酰)乙酰胺(8g7)的制备48.1 Preparation of intermediate 2-chloro-N-((4-(trifluoromethyl)oxazol-2-yl)carbamoyl)acetamide (8g7)
按照实施例1中步骤1.2的方法合成中间体8g7。1H NMR(400MHz,DMSO-d6)δ11.17(s,1H),11.08(s,1H),8.73(s,1H),4.48(s,2H).Intermediate 8g7 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.17(s,1H),11.08(s,1H),8.73(s,1H),4.48(s,2H).
48.2终产物I-9g7的制备48.2 Preparation of final product I-9g7
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8g7，制得化合物I-9g7。1H NMR(400MHz,DMSO-d6)δ11.55(s,1H),11.25(d,J＝45.6Hz,2H),9.66(s,1H),8.91-8.60(m,2H),7.76(s,2H),7.70-7.47(m,4H),7.37(d,J＝11.3Hz,1H),6.00(d,J＝10.6Hz,1H),4.53(d,J＝11.7Hz,1H),4.35(d,J＝11.5Hz,1H),3.11(s,3H).Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8g7, and prepare compound I-9g7. 1 H NMR (400MHz, DMSO-d 6 ) δ11.55 (s, 1H), 11.25 (d, J = 45.6Hz, 2H), 9.66 (s, 1H), 8.91-8.60 (m, 2H), 7.76 ( s,2H),7.70-7.47(m,4H),7.37(d,J＝11.3Hz,1H),6.00(d,J＝10.6Hz,1H),4.53(d,J＝11.7Hz,1H), 4.35(d,J＝11.5Hz,1H),3.11(s,3H).
实施例49Example 49
2-(3-(1,3-二甲基-1H-吡唑-5-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9g8)2-(3-(1,3-dimethyl-1H-pyrazol-5-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2 ,3-dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio (I-9g8)
49.1中间体2-氯-N-((1,3-二甲基-1H-吡唑-5-基)氨甲酰)乙酰胺(8g8)的制备49.1 Preparation of intermediate 2-chloro-N-((1,3-dimethyl-1H-pyrazol-5-yl)carbamoyl)acetamide (8g8)
按照实施例1中步骤1.2的方法合成中间体8g8。1H NMR(400MHz,DMSO-d6)δ11.19(s,1H),10.36(s,1H),6.16(s,1H),4.47(s,2H),2.16(s,3H).Intermediate 8g8 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.19(s,1H),10.36(s,1H),6.16(s,1H),4.47(s,2H),2.16(s,3H).
49.2终产物I-9g8的制备49.2 Preparation of final product I-9g8
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8g8，制得化合物I-9g8。ESI-MS:m/z 536.2[M+H]+.Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8g8, and prepare compound I-9g8. ESI-MS:m/z 536.2[M+H] + .
实施例50Example 50
2-(3-(1-甲基-1H-吡唑-5-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9g9)2-(3-(1-methyl-1H-pyrazol-5-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3- Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9g9)
50.1中间体2-氯-N-((1-甲基-1H-吡唑-5-基)氨甲酰)乙酰胺(8g9)的制备50.1 Preparation of intermediate 2-chloro-N-((1-methyl-1H-pyrazol-5-yl)carbamoyl)acetamide (8g9)
按照实施例1中步骤1.2的方法合成中间体I-8g9。1H NMR(400MHz,DMSO-d6)δ 11.14(s,1H),10.13(s,1H),7.36(s,1H),6.23(s,1H),4.44(s,2H),3.68(s,3H).Intermediate I-8g9 was synthesized according to the method of step 1.2 in Example 1. 1H NMR (400MHz, DMSO-d 6 )δ 11.14(s,1H),10.13(s,1H),7.36(s,1H),6.23(s,1H),4.44(s,2H),3.68(s,3H).
50.2终产物I-9g9的制备50.2 Preparation of final product I-9g9
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8g9，制得化合物I-9g9。ESI-MS:m/z 522.1[M+H]+.Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8g9, and prepare compound I-9g9. ESI-MS:m/z 522.1[M+H] + .
实施例51Example 51
2-(3-(5-甲基异噁唑-3-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9h1)2-(3-(5-methylisoxazol-3-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-benzo[e][1,4]diazepine -3-yl)Carbodithio (I-9h1)
51.1中间体2-氯-N-((5-甲基异噁唑-3-基)氨甲酰)乙酰胺(8h1)的制备51.1 Preparation of intermediate 2-chloro-N-((5-methylisoxazol-3-yl)carbamoyl)acetamide (8h1)
按照实施例1中步骤1.2的方法合成中间体8h1。1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),10.50(s,1H),6.58(d,J＝1.2Hz,1H),4.41(s,2H),2.38(d,J＝1.0Hz,3H).Intermediate 8h1 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.09 (s, 1H), 10.50 (s, 1H), 6.58 (d, J = 1.2Hz, 1H), 4.41 (s, 2H), 2.38 (d, J＝1.0Hz,3H).
51.2终产物I-9h1的制备51.2 Preparation of final product I-9h1
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8h1，制得化合物I-9h1。1H NMR(400MHz,DMSO-d6)δ11.52(d,J＝6.4Hz,1H),11.19(s,1H),10.66(s,1H),7.77-7.66(m,2H),7.62-7.57(m,2H),7.55(d,J＝7.0Hz,1H),7.51-7.45(m,2H),7.37-7.29(m,2H),6.58(d,J＝4.2Hz,1H),5.97(d,J＝6.5Hz,1H),4.27(s,2H),3.40(s,3H),2.38(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8h1, and prepare compound I-9h1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.52 (d, J = 6.4 Hz, 1H), 11.19 (s, 1H), 10.66 (s, 1H), 7.77-7.66 (m, 2H), 7.62- 7.57(m,2H),7.55(d,J＝7.0Hz,1H),7.51-7.45(m,2H),7.37-7.29(m,2H),6.58(d,J＝4.2Hz,1H),5.97 (d,J=6.5Hz,1H),4.27(s,2H),3.40(s,3H),2.38(s,3H) .
实施例52Example 52
2-(3-(1-甲基-1H-1,2,4-三唑-3-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9h2)2-(3-(1-methyl-1H-1,2,4-triazol-3-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-benzene Base-2,3-dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9h2)
52.1中间体2-氯-N-((1-甲基-1H-1,2,4-三唑-3-基)氨甲酰)乙酰胺(8h2)的制备52.1 Preparation of intermediate 2-chloro-N-((1-methyl-1H-1,2,4-triazol-3-yl)carbamoyl)acetamide (8h2)
按照实施例1中步骤1.2的方法合成中间体8h2。1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),10.30(s,1H),8.37(s,1H),4.45(s,2H),3.80(s,3H).Intermediate 8h2 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.08(s,1H),10.30(s,1H),8.37(s,1H),4.45(s,2H),3.80(s,3H).
52.2终产物I-9h2的制备52.2 Preparation of final product I-9h2
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8h2，制得化合物I-9h2。1H NMR(400MHz,DMSO-d6)δ11.46(s,1H),11.05(s,1H),10.38(s,1H),8.33(s,1H),7.75-7.66(m,2H),7.59(d,J＝7.7Hz,2H),7.54(d,J＝5.7Hz,1H),7.48(t,J＝6.8Hz,2H),7.33(d,J＝8.3Hz,2H),6.00-5.91(m,1H),4.27(s,2H),3.79(s,3H),3.39(s,3H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8h2, and prepare compound I-9h2. 1 H NMR (400MHz, DMSO-d 6 ) δ11.46(s,1H),11.05(s,1H),10.38(s,1H),8.33(s,1H),7.75-7.66(m,2H), 7.59(d,J＝7.7Hz,2H),7.54(d,J＝5.7Hz,1H),7.48(t,J＝6.8Hz,2H),7.33(d,J＝8.3Hz,2H),6.00- 5.91(m,1H),4.27(s,2H),3.79(s,3H),3.39(s,3H) .
实施例53Example 53
2-(吗啉-4-甲酰胺基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9i1)2-(morpholine-4-carboxamido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][ 1,4]diaza -3-yl)Carbodithio(I-9i1)
53.1中间体N-(2-氯乙酰基)吗啉-4-甲酰胺(8i1)的制备53.1 Preparation of intermediate N-(2-chloroacetyl)morpholine-4-carboxamide (8i1)
按照实施例1中步骤1.2的方法合成中间体8i1。1H NMR(400MHz,DMSO-d6)δ10.25(s,1H),4.54-4.42(m,2H),3.54(dq,J＝8.0,3.3,2.7Hz,4H),3.36(dd,J＝5.3,2.9Hz,4H).Intermediate 8i1 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.25 (s, 1H), 4.54-4.42 (m, 2H), 3.54 (dq, J = 8.0, 3.3, 2.7Hz, 4H), 3.36 (dd, J =5.3,2.9Hz,4H).
53.2终产物I-9i1的制备53.2 Preparation of final product I-9i1
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8i1，制得化合物I-9i1。1H NMR(400MHz,DMSO-d6)δ11.41(s,1H),10.16(s,1H),7.76-7.68(m,2H),7.59(d,J＝8.8Hz,2H),7.54(d,J＝5.3Hz,1H),7.51-7.44(m,2H),7.37-7.30(m,2H),5.96(s,1H),4.26 (s,2H),3.54(s,4H),3.38(s,3H),3.35-3.34(m,4H). Compound I-9i1 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8i1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.41 (s, 1H), 10.16 (s, 1H), 7.76-7.68 (m, 2H), 7.59 (d, J = 8.8Hz, 2H), 7.54 ( d,J＝5.3Hz,1H),7.51-7.44(m,2H),7.37-7.30(m,2H),5.96(s,1H),4.26 (s,2H),3.54(s,4H),3.38(s,3H),3.35-3.34(m,4H) .
实施例54Example 54
2-(4-甲氧基哌啶-1-甲酰胺基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9i2)2-(4-methoxypiperidine-1-carboxamido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzene And[e][1,4]diazapine -3-yl)Carbodithio(I-9i2)
54.1中间体N-(2-氯乙酰基)-4-甲氧基哌啶-1-甲酰胺(8i2)的制备54.1 Preparation of intermediate N-(2-chloroacetyl)-4-methoxypiperidine-1-carboxamide (8i2)
按照实施例1中步骤1.2的方法合成中间体8i2。1H NMR(400MHz,DMSO-d6)δ10.20(s,1H),4.47(s,2H),3.62(d,J＝5.7Hz,2H),3.41(d,J＝23.7Hz,2H),3.16(s,3H),1.79(s,2H),1.42(s,2H).Intermediate 8i2 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.20 (s, 1H), 4.47 (s, 2H), 3.62 (d, J = 5.7Hz, 2H), 3.41 (d, J = 23.7Hz, 2H) ,3.16(s,3H),1.79(s,2H),1.42(s,2H).
54.2终产物I-9i2的制备54.2 Preparation of final product I-9i2
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8i2，制得化合物I-9i2。1H NMR(400MHz,DMSO-d6)δ11.36(d,J＝6.3Hz,1H),10.07(s,1H),7.76-7.67(m,2H),7.58(d,J＝7.9Hz,2H),7.54(d,J＝6.3Hz,1H),7.47(t,J＝7.2Hz,2H),7.33(q,J＝7.6Hz,2H),5.96(d,J＝6.7Hz,1H),4.25(s,2H),3.38(s,3H),3.24(d,J＝1.2Hz,1H),3.22(s,3H),1.78(d,J＝6.6Hz,4H),1.44-1.30(m,4H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8i2, and prepare compound I-9i2. 1 H NMR (400MHz, DMSO-d 6 ) δ11.36 (d, J = 6.3 Hz, 1H), 10.07 (s, 1H), 7.76-7.67 (m, 2H), 7.58 (d, J = 7.9 Hz, 2H),7.54(d,J＝6.3Hz,1H),7.47(t,J＝7.2Hz,2H),7.33(q,J＝7.6Hz,2H),5.96(d,J＝6.7Hz,1H) ,4.25(s,2H),3.38(s,3H),3.24(d,J＝1.2Hz,1H),3.22(s,3H),1.78(d,J＝6.6Hz,4H),1.44-1.30( m,4H) .
实施例55Example 55
2-氧代-2-(1,4-二氧杂-8-氮杂螺环[4.5]癸烷-8-甲酰胺基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9i3)2-Oxo-2-(1,4-dioxa-8-azaspiro[4.5]decane-8-carboxamido)ethyl(1-methyl-2-oxo-5-benzene Base-2,3-dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9i3)
55.1中间体N-(2-氯乙酰基)-1,4-二氧杂-8-氮杂螺环[4.5]癸烷-8-甲酰胺(8i3)的制备55.1 Preparation of intermediate N-(2-chloroacetyl)-1,4-dioxa-8-azaspiro[4.5]decane-8-carboxamide (8i3)
按照实施例1中步骤1.2的方法合成中间体8i3。1H NMR(400MHz,DMSO-d6)δ10.23(s,1H),4.47(s,2H),3.90(d,J＝6.0Hz,4H),3.49-3.36(m,4H),1.67-1.58(m,4H).Intermediate 8i3 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.23 (s, 1H), 4.47 (s, 2H), 3.90 (d, J = 6.0Hz, 4H), 3.49-3.36 (m, 4H), 1.67- 1.58(m,4H).
55.2终产物I-9i3的制备55.2 Preparation of final product I-9i3
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8i3，制得化合物I-9i3。1H NMR(400MHz,DMSO-d6)δ11.36(d,J＝6.5Hz,1H),10.14(s,1H),7.75-7.67(m,2H),7.58(d,J＝8.0Hz,2H),7.53(d,J＝7.0Hz,1H),7.47(t,J＝7.3Hz,2H),7.32(t,J＝8.3Hz,2H),5.96(d,J＝6.4Hz,1H),4.25(s,2H),3.87(s,4H),3.42(d,J＝4.4Hz,4H),3.38(s,3H),1.61-1.56(m,4H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8i3, and prepare compound I-9i3. 1 H NMR (400MHz, DMSO-d 6 ) δ11.36 (d, J = 6.5Hz, 1H), 10.14 (s, 1H), 7.75-7.67 (m, 2H), 7.58 (d, J = 8.0Hz, 2H),7.53(d,J＝7.0Hz,1H),7.47(t,J＝7.3Hz,2H),7.32(t,J＝8.3Hz,2H),5.96(d,J＝6.4Hz,1H) ,4.25(s,2H),3.87(s,4H),3.42(d,J＝4.4Hz,4H),3.38(s,3H),1.61-1.56(m,4H) .
实施例56Example 56
2-氧代-2-(3-(四氢-2H-吡喃-4-基)脲基)乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9i4)2-Oxo-2-(3-(tetrahydro-2H-pyran-4-yl)ureido)ethyl(1-methyl-2-oxo-5-phenyl-2,3-dihydro -1H-benzo[e][1,4]diazepine -3-yl)Carbodithio(I-9i4)
56.1中间体2-氯-N-((四氢-2H-吡喃-4-基)氨甲酰)乙酰胺(8i4)的制备56.1 Preparation of intermediate 2-chloro-N-((tetrahydro-2H-pyran-4-yl)carbamoyl)acetamide (8i4)
按照实施例1中步骤1.2的方法合成中间体8i4。1H NMR(400MHz,DMSO-d6)δ10.60(s,1H),8.05(s,1H),4.27(s,2H),3.79(d,J＝12.7Hz,4H),1.77(d,J＝12.6Hz,2H),1.62(d,J＝21.1Hz,1H),1.45(tt,J＝11.0,5.8Hz,2H).Intermediate 8i4 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.60 (s, 1H), 8.05 (s, 1H), 4.27 (s, 2H), 3.79 (d, J = 12.7Hz, 4H), 1.77 (d, J＝12.6Hz,2H),1.62(d,J＝21.1Hz,1H),1.45(tt,J＝11.0,5.8Hz,2H).
56.2终产物I-9i4的制备56.2 Preparation of final product I-9i4
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8i4，制得化合物I-9i4。1H NMR(400MHz,DMSO-d6)δ11.45(d,J＝6.1Hz,1H),10.54(s,1H),8.14(s,1H), 7.76-7.68(m,2H),7.62-7.53(m,3H),7.52-7.45(m,2H),7.37-7.29(m,2H),5.97(d,J＝8.2Hz,1H),4.15(s,2H),3.85-3.74(m,4H),3.39(s,3H),1.98(d,J＝8.1Hz,1H),1.76(d,J＝4.7Hz,2H),1.61(s,2H). Synthesize according to the method of step 1.3 in Example 1, replace the raw materials with the same equivalent of 8i4, and prepare compound I-9i4. 1 H NMR (400MHz, DMSO-d 6 ) δ11.45 (d, J = 6.1 Hz, 1H), 10.54 (s, 1H), 8.14 (s, 1H), 7.76-7.68(m,2H),7.62-7.53(m,3H),7.52-7.45(m,2H),7.37-7.29(m,2H),5.97(d,J＝8.2Hz,1H),4.15( s,2H),3.85-3.74(m,4H),3.39(s,3H),1.98(d,J＝8.1Hz,1H),1.76(d,J＝4.7Hz,2H),1.61(s,2H ) .
实施例57Example 57
2-(3-((3s,5s,7s)-金刚烷-1-基)脲基)-2-氧代乙基(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-9i5)2-(3-((3s,5s,7s)-adamantan-1-yl)ureido)-2-oxoethyl(1-methyl-2-oxo-5-phenyl-2,3 -Dihydro-1H-benzo[e][1,4]diaza -3-yl)Carbodithio(I-9i5)
57.1中间体N-(((3s,5s,7s)-金刚烷-1-基)氨甲酰)-2-氯乙酰胺(8i5)的制备57.1 Preparation of intermediate N-(((3s,5s,7s)-adamantane-1-yl)carbamoyl)-2-chloroacetamide (8i5)
按照实施例1中步骤1.2的方法合成中间体8i5。1H NMR(400MHz,DMSO-d6)δ10.41(s,1H),8.01(s,1H),4.25(q,J＝6.4,5.0Hz,2H),2.04(s,3H),1.97-1.91(m,6H),1.63(d,J＝8.7Hz,6H).Intermediate 8i5 was synthesized according to the method of step 1.2 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ10.41 (s, 1H), 8.01 (s, 1H), 4.25 (q, J = 6.4, 5.0Hz, 2H), 2.04 (s, 3H), 1.97- 1.91(m,6H),1.63(d,J＝8.7Hz,6H).
57.2终产物I-9i5的制备57.2 Preparation of final product I-9i5
按照实施例1中步骤1.3的方法合成，将原料替换成相同当量的8i5，制得化合物I-9i5。1H NMR(400MHz,DMSO-d6)δ11.43(d,J＝19.5Hz,1H),10.35(s,1H),8.11(s,1H),7.77-7.67(m,2H),7.63-7.53(m,3H),7.48(t,J＝7.4Hz,2H),7.38-7.29(m,2H),5.97(d,J＝6.3Hz,1H),4.13(s,2H),3.40(s,3H),2.02(s,3H),1.92(d,J＝9.3Hz,6H),1.62(s,6H). Compound I-9i5 was synthesized according to the method of step 1.3 in Example 1, replacing the raw materials with the same equivalent of 8i5. 1 H NMR (400MHz, DMSO-d 6 ) δ11.43 (d, J = 19.5Hz, 1H), 10.35 (s, 1H), 8.11 (s, 1H), 7.77-7.67 (m, 2H), 7.63- 7.53(m,3H),7.48(t,J＝7.4Hz,2H),7.38-7.29(m,2H),5.97(d,J＝6.3Hz,1H),4.13(s,2H),3.40(s ,3H),2.02(s,3H),1.92(d,J=9.3Hz,6H),1.62(s,6H) .
实施例58Example 58
2-氧代-2-(3-(4-(三氟甲氧基)苯基)脲基)乙基(2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-10a1)2-Oxo-2-(3-(4-(trifluoromethoxy)phenyl)ureido)ethyl(2-oxo-5-phenyl-2,3-dihydro-1H-benzo [e][1,4]diazapine -3-yl)Carbodithio(I-10a1)
58.1终产物I-10a1的制备58.1 Preparation of final product I-10a1
按照实施例1中步骤1.3的方法合成，制得化合物I-10a1。1H NMR(400MHz,DMSO-d6)δ11.49(s,1H),11.00(s,2H),10.42(s,1H),7.65(dd,J＝8.8,3.4Hz,3H),7.57-7.51(m,3H),7.48(d,J＝7.3Hz,2H),7.36-7.29(m,4H),7.26(d,J＝6.9Hz,1H),5.91(d,J＝5.5Hz,1H),4.28(s,2H). Compound I-10a1 was synthesized according to the method of step 1.3 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.49 (s, 1H), 11.00 (s, 2H), 10.42 (s, 1H), 7.65 (dd, J = 8.8, 3.4Hz, 3H), 7.57- 7.51(m,3H),7.48(d,J＝7.3Hz,2H),7.36-7.29(m,4H),7.26(d,J＝6.9Hz,1H),5.91(d,J＝5.5Hz,1H ),4.28(s,2H) .
实施例59Example 59
2-(3-(4-(二甲氨基)苯基)脲基)-2-氧乙基(2-氧基-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-10a2)2-(3-(4-(Dimethylamino)phenyl)ureido)-2-oxyethyl(2-oxy-5-phenyl-2,3-dihydro-1H-benzo[e] [1,4]Diazepine -3-yl)Carbodithio(I-10a2)
59.1终产物I-10a2的制备59.1 Preparation of final product I-10a2
按照实施例1中步骤1.3的方法合成，制得化合物I-10a2。1H NMR(400MHz,DMSO-d6)δ11.45(s,1H),10.99(s,1H),10.80(s,1H),10.04(s,1H),7.65(t,J＝7.7Hz,1H),7.54(d,J＝7.8Hz,3H),7.49(d,J＝7.2Hz,2H),7.30(td,J＝14.6,14.1,8.5Hz,5H),6.69(d,J＝8.5Hz,2H),5.92(d,J＝6.4Hz,1H),4.25(s,2H),2.85(d,J＝4.0Hz,6H). Compound I-10a2 was synthesized according to the method of step 1.3 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.45 (s, 1H), 10.99 (s, 1H), 10.80 (s, 1H), 10.04 (s, 1H), 7.65 (t, J = 7.7Hz, 1H),7.54(d,J＝7.8Hz,3H),7.49(d,J＝7.2Hz,2H),7.30(td,J＝14.6,14.1,8.5Hz,5H),6.69(d,J＝8.5 Hz,2H),5.92(d,J＝6.4Hz,1H),4.25(s,2H),2.85(d,J＝4.0Hz,6H) .
实施例60Example 60
2-氧代-2-(3-(2-(三氟甲基)苯基)脲基)乙基(2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-10a3)2-Oxo-2-(3-(2-(trifluoromethyl)phenyl)ureido)ethyl(2-oxo-5-phenyl-2,3-dihydro-1H-benzo[ e][1,4]diaza -3-yl)Carbodithio(I-10a3)
60.1终产物I-10a3的制备60.1 Preparation of final product I-10a3
按照实施例1中步骤1.3的方法合成，制得化合物I-10a3。1H NMR(400MHz, DMSO-d6)δ11.46(s,1H),11.20(s,1H),10.98(s,1H),10.75(s,1H),8.12(d,J＝8.1Hz,1H),7.69(dq,J＝20.2,7.6Hz,3H),7.54(d,J＝7.3Hz,3H),7.48(t,J＝7.7Hz,2H),7.31(td,J＝19.1,18.1,8.1Hz,4H),5.91(d,J＝6.6Hz,1H),4.30(d,J＝5.1Hz,2H). Compound I-10a3 was synthesized according to the method of step 1.3 in Example 1. 1 H NMR (400MHz, DMSO-d 6 )δ11.46(s,1H),11.20(s,1H),10.98(s,1H),10.75(s,1H),8.12(d,J＝8.1Hz,1H),7.69(dq ,J＝20.2,7.6Hz,3H),7.54(d,J＝7.3Hz,3H),7.48(t,J＝7.7Hz,2H),7.31(td,J＝19.1,18.1,8.1Hz,4H) ,5.91(d,J＝6.6Hz,1H),4.30(d,J＝5.1Hz,2H) .
实施例61Example 61
2-(3-(5-氯-2-硝基苯基)脲基)-2-氧代乙基(2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-10a4)2-(3-(5-chloro-2-nitrophenyl)ureido)-2-oxoethyl(2-oxo-5-phenyl-2,3-dihydro-1H-benzo[ e][1,4]diaza -3-yl)Carbodithio(I-10a4)
61.1终产物I-10a4的制备61.1 Preparation of final product I-10a4
按照实施例1中步骤1.3的方法合成，制得化合物I-10a4。1H NMR(400MHz,DMSO-d6)δ12.06(s,1H),11.48(d,J＝6.6Hz,1H),11.30(s,1H),10.96(s,1H),8.57(d,J＝2.3Hz,1H),8.16(d,J＝8.9Hz,1H),7.60(t,J＝7.5Hz,1H),7.49(d,J＝7.4Hz,3H),7.46-7.40(m,2H),7.34(dd,J＝8.9,2.3Hz,1H),7.31-7.24(m,2H),7.20(t,J＝7.6Hz,1H),5.86(d,J＝6.7Hz,1H),4.24(d,J＝4.6Hz,2H). Compound I-10a4 was synthesized according to the method of step 1.3 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ12.06 (s, 1H), 11.48 (d, J = 6.6Hz, 1H), 11.30 (s, 1H), 10.96 (s, 1H), 8.57 (d, J＝2.3Hz,1H),8.16(d,J＝8.9Hz,1H),7.60(t,J＝7.5Hz,1H),7.49(d,J＝7.4Hz,3H),7.46-7.40(m, 2H),7.34(dd,J＝8.9,2.3Hz,1H),7.31-7.24(m,2H),7.20(t,J＝7.6Hz,1H),5.86(d,J＝6.7Hz,1H), 4.24(d,J=4.6Hz,2H) .
实施例62Example 62
2-氧代-2-(3-(吡啶-2-基)脲基)乙基(2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-10a5)2-Oxo-2-(3-(pyridin-2-yl)ureido)ethyl(2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1, 4]Diazepine -3-yl)Carbodithio(I-10a5)
62.1终产物I-10a5的制备62.1 Preparation of final product I-10a5
按照实施例1中步骤1.3的方法合成，制得化合物I-10a5。1H NMR(400MHz,DMSO-d6)δ11.49(s,1H),11.16(s,1H),11.04-10.96(m,1H),10.71(s,1H),8.31(s,1H),7.96(s,1H),7.82(s,1H),7.66(s,1H),7.58-7.45(m,5H),7.36-7.22(m,3H),7.14(s,1H),5.92(s,1H),4.30(s,2H). Compound I-10a5 was synthesized according to the method of step 1.3 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.49(s,1H),11.16(s,1H),11.04-10.96(m,1H),10.71(s,1H),8.31(s,1H), 7.96(s,1H),7.82(s,1H),7.66(s,1H),7.58-7.45(m,5H),7.36-7.22(m,3H),7.14(s,1H),5.92(s, 1H),4.30(s,2H) .
实施例63Example 63
2-(吗啉-4-甲酰胺基)-2-氧代乙基(2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)碳二硫代(I-10a6)2-(morpholine-4-carboxamido)-2-oxoethyl(2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]di Aza -3-yl)Carbodithio(I-10a6)
63.1终产物I-10a6的制备63.1 Preparation of final product I-10a6
按照实施例1中步骤1.3的方法合成，制得化合物I-10a6。1H NMR(400MHz,DMSO-d6)δ11.33(s,1H),10.97(s,1H),10.14(s,1H),7.64(s,1H),7.49(s,3H),7.45(d,J＝7.5Hz,2H),7.31(s,2H),7.22(t,J＝7.6Hz,1H),5.83(s,1H),4.27(s,2H),3.53(s,4H),3.35(s,4H). Compound I-10a6 was synthesized according to the method of step 1.3 in Example 1. 1 H NMR (400MHz, DMSO-d 6 ) δ11.33(s,1H),10.97(s,1H),10.14(s,1H),7.64(s,1H),7.49(s,3H),7.45( d,J＝7.5Hz,2H),7.31(s,2H),7.22(t,J＝7.6Hz,1H),5.83(s,1H),4.27(s,2H),3.53(s,4H), 3.35(s,4H) .
实施例64Example 64
2-(3-(3-氯-5-(三氟甲基)苯基)脲基)-N-(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)噁唑-5-甲酰胺(I-11a1)2-(3-(3-chloro-5-(trifluoromethyl)phenyl)ureido)-N-(1-methyl-2-oxo-5-phenyl-2,3-dihydro- 1H-benzo[e][1,4]diazepine -3-yl)oxazole-5-carboxamide (I-11a1)
64.1中间体2-(3-(3-氯-5-(三氟甲基)苯基)脲基)噁唑-5-羧酸(8j1)的制备64.1 Preparation of intermediate 2-(3-(3-chloro-5-(trifluoromethyl)phenyl)ureido)oxazole-5-carboxylic acid (8j1)
操作与8j2的合成相同。1H NMR(400MHz,DMSO-d6)δ13.44(s,1H),11.44(s,1H),10.29(s,1H),8.08(s,1H),7.82(s,1H),7.73(s,1H),7.65(s,1H).The operation is the same as the synthesis of 8j2. 1 H NMR (400MHz, DMSO-d 6 ) δ13.44(s,1H),11.44(s,1H),10.29(s,1H),8.08(s,1H),7.82(s,1H),7.73( s,1H),7.65(s,1H).
64.2终产物I-11a1的制备
64.2 Preparation of final product I-11a1
称取化合物8j1 0.132g(0.38mmol)，EDCI 0.07g(0.38mmol),HOBT 0.06g(0.45mmol)置于100mL圆底烧瓶中，加入氮甲基吗啉1mL和20mL二氯甲烷，常温下搅拌30min后，将化合物7 0.1g(0.38mmol)加入反应体系，常温下反应过夜。将反应体系真空浓缩后，过色谱柱得到终产物I-11a1，产率50％。1H NMR(400MHz,DMSO-d6)δ10.39(s,1H),9.41(s,1H),8.11(s,2H),7.72(dq,J＝25.7,9.3Hz,5H),7.57(d,J＝7.6Hz,2H),7.52(d,J＝6.4Hz,1H),7.46(d,J＝7.4Hz,2H),7.34(s,2H),5.45(d,J＝7.7Hz,1H),3.39(s,3H). Weigh 0.132g (0.38mmol) of compound 8j1, 0.07g (0.38mmol) EDCI, and 0.06g (0.45mmol) HOBT into a 100mL round-bottom flask, add 1mL nitrogen methylmorpholine and 20mL dichloromethane, and stir at room temperature. After 30 minutes, 0.1g (0.38mmol) of compound 7 was added to the reaction system, and the reaction was carried out overnight at room temperature. After the reaction system was concentrated in vacuo, the final product I-11a1 was obtained through a chromatography column with a yield of 50%. 1 H NMR (400MHz, DMSO-d 6 ) δ10.39 (s, 1H), 9.41 (s, 1H), 8.11 (s, 2H), 7.72 (dq, J = 25.7, 9.3Hz, 5H), 7.57 ( d,J＝7.6Hz,2H),7.52(d,J＝6.4Hz,1H),7.46(d,J＝7.4Hz,2H),7.34(s,2H),5.45(d,J＝7.7Hz, 1H),3.39(s,3H) .
实施例65Example 65
3-(3-(3-氯-5-(三氟甲基)苯基)脲基)-N-(1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)苯甲酰胺(I-11a2)3-(3-(3-chloro-5-(trifluoromethyl)phenyl)ureido)-N-(1-methyl-2-oxo-5-phenyl-2,3-dihydro- 1H-benzo[e][1,4]diazepine -3-yl)benzamide (I-11a2)
65.1中间体8j2的制备
65.1 Preparation of Intermediate 8j2
称取3-氨基苯甲酸乙酯0.5g(3mmol,)与三乙胺0.7mL(4.5mmol)溶于100mL圆底烧瓶中，加入二氯甲烷溶解，在常温下加入4-氯-3-三氟甲基异氰酸苯酯0.45g(2mmol)，反应过夜。反应中固体析出，抽滤所得化合物加入2N NaOH:甲醇＝1:1的溶剂20mL，常温下反应3h后，在体系中加入过量盐酸调PH至酸性，真空旋干有机溶剂后，加入EA萃取三次。无水硫酸钠干燥有机相，真空浓缩后，硅胶柱层析纯化得8j2，白色固体，产率50％。1H NMR(400MHz,DMSO-d6)δ12.93(s,1H),9.19(s,1H),9.06(s,1H),8.16-8.05(m,2H),7.70-7.51(m,4H),7.40(d,J＝7.7Hz,1H).Weigh 0.5g (3mmol,) of 3-aminobenzoic acid ethyl ester and 0.7mL (4.5mmol,) of triethylamine and dissolve it in a 100mL round-bottomed flask. Add methylene chloride to dissolve. Add 4-chloro-3-triethylamine at room temperature. Fluoromethyl phenyl isocyanate 0.45g (2mmol), react overnight. During the reaction, a solid precipitated. The obtained compound was filtered and 20 mL of a solvent of 2N NaOH: methanol = 1:1 was added. After reacting for 3 hours at room temperature, excess hydrochloric acid was added to the system to adjust the pH to acidic. After the organic solvent was dried in a vacuum, EA was added for extraction three times. . The organic phase was dried over anhydrous sodium sulfate, concentrated in vacuo, and purified by silica gel column chromatography to obtain 8j2 as a white solid with a yield of 50%. 1 H NMR (400MHz, DMSO-d 6 ) δ12.93(s,1H),9.19(s,1H),9.06(s,1H),8.16-8.05(m,2H),7.70-7.51(m,4H ),7.40(d,J＝7.7Hz,1H).
65.2终产物I-11a2的制备65.2 Preparation of final product I-11a2
操作与11a1的合成相同。1H NMR(400MHz,DMSO-d6)δ9.66(s,1H),9.47(d,J＝7.9Hz,1H),9.36(s,1H),8.11(s,1H),7.94(s,1H),7.68(dt,J＝20.0,9.7Hz,5H),7.62-7.56(m,3H),7.51(d,J＝7.0Hz,1H),7.49-7.33(m,5H),5.52(d,J＝7.7Hz,1H),3.40(s,3H). The operation is the same as the synthesis of 11a1. 1 H NMR (400MHz, DMSO-d 6 ) δ9.66 (s, 1H), 9.47 (d, J = 7.9Hz, 1H), 9.36 (s, 1H), 8.11 (s, 1H), 7.94 (s, 1H),7.68(dt,J＝20.0,9.7Hz,5H),7.62-7.56(m,3H),7.51(d,J＝7.0Hz,1H),7.49-7.33(m,5H),5.52(d ,J＝7.7Hz,1H),3.40(s,3H) .
实施例66Example 66
5-氯-N-(4-((1-甲基-2-氧代-5-苯基-2,3-二氢-1H-苯并[e][1,4]二氮杂-3-基)氨基)-4-氧代丁基)-2-硝基苯甲酰胺(I-11a3)5-Chloro-N-(4-((1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]diaza -3-yl)amino)-4-oxobutyl)-2-nitrobenzamide (I-11a3)
66.1中间体8j3的制备
66.1 Preparation of Intermediate 8j3
称取3-氯-2-硝基苯甲酸0.3g(1.5mmol)，EDCI 0.38g(3.0mmol),HOBT 0.85g(2.2 mmol)置于100mL圆底烧瓶中，加入氮甲基吗啉1mL和20mL二氯甲烷，常温下搅拌30min后，将4-氨基丁酸乙酯0.25g(1.5mmol)加入反应体系，常温下反应过夜。将反应体系真空浓缩后得到酰胺缩合产物。1H NMR(400MHz,DMSO-d6)δ8.77(d,J＝5.7Hz,1H),8.08(dd,J＝8.6,1.8Hz,1H),7.81-7.71(m,2H),4.07(tt,J＝8.0,4.1Hz,2H),3.23(t,J＝6.6Hz,2H),2.39(t,J＝7.5Hz,2H),1.76(p,J＝7.1Hz,2H),1.19(td,J＝7.1,1.7Hz,3H).将该产物加入到甲醇：NaOH(2N)水溶液＝1:1的20mL溶剂中，常温下反应2h，用2N HCl中和后，用乙酸乙酯萃取，用无水硫酸钠干燥有机相，真空浓缩，柱层析得到化合物8j3，白色固体，收率48％。1H NMR(400MHz,DMSO-d6)δ12.11(s,1H),8.77(s,1H),8.08(s,1H),7.75(s,2H),3.22(s,2H),2.31(s,2H),1.72(s,2H).Weigh 0.3g (1.5mmol) of 3-chloro-2-nitrobenzoic acid, 0.38g (3.0mmol) of EDCI, 0.85g (2.2 mmol) into a 100 mL round-bottomed flask, add 1 mL of nitrogen methylmorpholine and 20 mL of methylene chloride, stir for 30 min at room temperature, add 0.25 g (1.5 mmol) of 4-aminobutyric acid ethyl ester to the reaction system, and react at room temperature. overnight. The reaction system was concentrated in vacuo to obtain the amide condensation product. 1 H NMR (400MHz, DMSO-d 6 ) δ8.77 (d, J = 5.7 Hz, 1H), 8.08 (dd, J = 8.6, 1.8 Hz, 1H), 7.81-7.71 (m, 2H), 4.07 ( tt,J＝8.0,4.1Hz,2H),3.23(t,J＝6.6Hz,2H),2.39(t,J＝7.5Hz,2H),1.76(p,J＝7.1Hz,2H),1.19( td, J = 7.1, 1.7Hz, 3H). Add the product to 20mL solvent of methanol: NaOH (2N) aqueous solution = 1:1, react at room temperature for 2h, neutralize with 2N HCl, and extract with ethyl acetate , dried the organic phase with anhydrous sodium sulfate, concentrated in vacuo, and obtained compound 8j3 as a white solid by column chromatography, with a yield of 48%. 1 H NMR (400MHz, DMSO-d 6 ) δ12.11(s,1H),8.77(s,1H),8.08(s,1H),7.75(s,2H),3.22(s,2H),2.31( s,2H),1.72(s,2H).
66.2终产物I-11a3的制备66.2 Preparation of final product I-11a3
操作与I-11a1的合成相同。1H NMR(400MHz,DMSO-d6)δ9.14(d,J＝12.3Hz,1H),8.74(s,1H),8.06(d,J＝9.1Hz,1H),7.80-7.63(m,4H),7.51(d,J＝10.2Hz,3H),7.48-7.41(m,2H),7.36-7.27(m,2H),5.31(d,J＝8.3Hz,1H),3.36(s,3H),3.24(d,J＝10.8Hz,2H),2.35(t,J＝7.3Hz,2H),1.81-1.69(m,2H). The operation is the same as for the synthesis of I-11a1. 1 H NMR (400MHz, DMSO-d 6 ) δ9.14 (d, J = 12.3 Hz, 1H), 8.74 (s, 1H), 8.06 (d, J = 9.1 Hz, 1H), 7.80-7.63 (m, 4H),7.51(d,J＝10.2Hz,3H),7.48-7.41(m,2H),7.36-7.27(m,2H),5.31(d,J＝8.3Hz,1H),3.36(s,3H ),3.24(d,J＝10.8Hz,2H),2.35(t,J＝7.3Hz,2H),1.81-1.69(m,2H) .
实施例67Example 67
本发明所述的1，4-苯二氮类化合物对ANXA3蛋白的结合活性测试。1,4-Benzodiazepine according to the present invention Binding activity test of similar compounds to ANXA3 protein.
实验方法：采用表面等离子共振法测定化合物式I对ANXA3蛋白的结合活性。Experimental method: Surface plasmon resonance method was used to determine the binding activity of compound formula I to ANXA3 protein.
实验主要试剂：重组ANXA3蛋白(由大肠杆菌系统表达纯化)、乙醇胺(美国Cytiva公司)、CM5芯片(美国Cytiva公司)、氨基偶联试剂盒(美国Cytiva公司)、Biacore T200生物大分子相互作用仪(美国通用电气公司)。Main reagents in the experiment: recombinant ANXA3 protein (expressed and purified by E. coli system), ethanolamine (U.S. Cytiva Company), CM5 chip (U.S. Cytiva Company), amino coupling kit (U.S. Cytiva Company), Biacore T200 biological macromolecule interaction instrument (General Electric Company, USA).
实验步骤：使用Biacore T200仪器进行测试，将ANXA3蛋白偶联于CM5传感器芯片中，选用PBS作为流动相，将化合物式I溶液流经芯片表面，流速为30μL/min，结合时间为30s，解离时间为45s，记录信号变化，采用Biacore T200软件对数据进行拟合分析，以计算出化合物式I与ANXA3蛋白的亲和力(KD)。Experimental steps: Use Biacore T200 instrument for testing, couple the ANXA3 protein to the CM5 sensor chip, use PBS as the mobile phase, flow the solution of compound formula I through the chip surface, the flow rate is 30 μL/min, the binding time is 30s, and dissociate The time is 45 s, the signal changes are recorded, and Biacore T200 software is used to perform fitting analysis on the data to calculate the affinity (K D ) of compound formula I and ANXA3 protein.
实施例68Example 68
本发明所述的1，4-苯二氮类化合物对ANXA3蛋白的降解活性测试。1,4-Benzodiazepine according to the present invention Test of the degradation activity of similar compounds on ANXA3 protein.
实验方法：采用蛋白免疫印迹法测定化合物式I对ANXA3蛋白的降解活性。Experimental method: The protein immunoblotting method was used to determine the degradation activity of compound formula I on ANXA3 protein.
细胞培养：高表达ANXA3的人源TNBC细胞MDA-MB-231在10％血清、100U/mL青霉素-链霉素的DMEM培养基中培养。培养条件：37℃，5％CO2。Cell culture: Human TNBC cells MDA-MB-231, which highly expresses ANXA3, were cultured in DMEM medium with 10% serum and 100 U/mL penicillin-streptomycin. Culture conditions: 37°C, 5% CO 2 .
实验主要试剂：蛋白酶抑制剂(瑞士Roche公司)、BCA蛋白定量检测试剂盒(美国Thermo Fisher Scientific公司)、ECL显色液(WBKLS0500、美国Milipore公司)、Tubulin抗体(HC101-02、北京全式金生物公司)、ANXA3抗体(11804-1-AP、美国Proteinteach公司)、二抗(HS201-01,HS101-01、北京全式金生物公司)。Main reagents in the experiment: protease inhibitor (Swiss Roche Company), BCA protein quantitative detection kit (American Thermo Fisher Scientific Company), ECL chromogenic solution (WBKLS0500, American Milipore Company), Tubulin antibody (HC101-02, Beijing Full Gold Biological Company), ANXA3 antibody (11804-1-AP, Proteinteach Company of the United States), secondary antibodies (HS201-01, HS101-01, Beijing Quanshijin Biotechnology Company).
实验步骤：将1.5*104个细胞接种于6cm培养皿中，24h后，将化合物式I溶液加入细胞作为实验组。将含0.1％DMSO的培养液加入细胞作为对照组。孵育24h后，收集细胞，加入RIAP溶液于冰上裂解30分钟，然后在4℃，12000转离心15分钟后收集上清。 BCA试剂盒检测总蛋白浓度，总蛋白上样量为50μg，浓缩胶电压电泳80V，分离胶电泳电压120V，然后110V恒压转膜90分钟，5％脱脂牛奶室温封闭1h，加入一抗4℃过夜，用TBST清洗，加入辣根过氧化物酶标记的IgG二抗，室温孵育1h，用TBST清洗，将ECL显色液均匀滴在PVDF膜上显影，使用Image J软件分析蛋白条带，计算半数降解浓度(DC50)。Experimental steps: 1.5*10 4 cells were seeded in a 6cm culture dish. After 24 hours, compound formula I solution was added to the cells to form the experimental group. Culture medium containing 0.1% DMSO was added to the cells as a control group. After incubation for 24 hours, cells were collected, RIAP solution was added and lysed on ice for 30 minutes, and then centrifuged at 12,000 rpm for 15 minutes at 4°C and the supernatant was collected. The BCA kit detects the total protein concentration. The total protein loading amount is 50 μg. The stacking gel electrophoresis voltage is 80V and the separation gel electrophoresis voltage is 120V. Then the film is transferred at a constant voltage of 110V for 90 minutes. Blocked with 5% skim milk at room temperature for 1 hour. Add the primary antibody at 4°C. Leave overnight, wash with TBST, add horseradish peroxidase-labeled IgG secondary antibody, incubate at room temperature for 1 hour, wash with TBST, evenly drop ECL chromogenic solution on the PVDF membrane for development, use Image J software to analyze protein bands, and calculate Half-degradation concentration (DC 50 ).
实施例69Example 69
本发明所述的1，4-苯二氮类化合物体外抑制肿瘤细胞活力实验。1,4-Benzodiazepine according to the present invention In vitro experiments on the inhibition of tumor cell viability by similar compounds.
细胞培养：与实施例68相同。Cell culture: Same as Example 68.
实验主要试剂：MTT(美国sigma公司)、胎牛血清(澳洲Ausbian公司)、DMEM培养基(美国Gibco公司)。Main reagents in the experiment: MTT (American Sigma Company), fetal calf serum (Ausbian Company, Australia), DMEM culture medium (American Gibco Company).
实验步骤：将5000个细胞传入96孔板中，24h后，加入化合物式I作为实验组。将含0.1％DMSO的培养基作为对照组。将未接种细胞孔作为空白组。孵育72h后，每孔加入100μL的MTT/PBS溶液，孵育4h。吸去上清液，加入150μL的DMSO，测定96孔板在490nm处的吸光值(OD)，采用GraphPad Prism 6软件计算化合物式I的半数抑制浓度(IC50)。其他肿瘤细胞(人宫颈癌细胞HeLa、人卵巢癌细胞A2780和人结肠癌细胞HCT116)活性测定方法与MDA-MB-231类似。Experimental steps: 5000 cells were transferred into a 96-well plate. After 24 hours, compound formula I was added as the experimental group. The medium containing 0.1% DMSO was used as a control group. The unseeded cell wells were used as blank groups. After incubation for 72 h, 100 μL of MTT/PBS solution was added to each well and incubated for 4 h. Aspirate the supernatant, add 150 μL of DMSO, measure the absorbance value (OD) of the 96-well plate at 490 nm, and use GraphPad Prism 6 software to calculate the half inhibitory concentration (IC 50 ) of compound formula I. The activity determination method of other tumor cells (human cervical cancer cell HeLa, human ovarian cancer cell A2780 and human colon cancer cell HCT116) is similar to MDA-MB-231.
实施例70Example 70
本发明所述的化合物I-9a19体外抑制肿瘤细胞克隆形成实验。Compound I-9a19 of the present invention inhibits tumor cell colony formation in vitro.
细胞培养：与实施例68相同。Cell culture: Same as Example 68.
实验主要试剂：多聚甲醛(国药集团化学试剂有限公司)、结晶紫(百灵威科技有限公司)。Main reagents in the experiment: paraformaldehyde (Sinopharm Chemical Reagent Co., Ltd.), crystal violet (Bailingwei Technology Co., Ltd.).
实验步骤：将2000个细胞传入6孔板，24h后，将2.5μM的I-9a19溶液加入细胞作为实验组。将含0.1％DMSO的培养液加入细胞作为对照组。每3天更换含化合物的培养液，培养2～3周后，当对照组出现可见克隆时，终止培养，加多聚甲醛固定15min。然后加结晶紫染色30min，然后清洗，晾干并拍照，计算克隆形成率。克隆形成率＝(克隆数/接种细胞数)×100％Experimental steps: 2000 cells were transferred into a 6-well plate. After 24 hours, 2.5 μM I-9a19 solution was added to the cells to form the experimental group. Culture medium containing 0.1% DMSO was added to the cells as a control group. The culture medium containing the compound was replaced every 3 days. After 2 to 3 weeks of culture, when visible colonies appeared in the control group, the culture was terminated and paraformaldehyde was added for fixation for 15 minutes. Then add crystal violet to stain for 30 minutes, then wash, dry and take photos to calculate the colony formation rate. Clone formation rate = (number of clones/number of inoculated cells) × 100%
实施例71Example 71
本发明所述的化合物I-9a19体外抑制肿瘤细胞迁移能力实验。Experiment on the ability of compound I-9a19 to inhibit tumor cell migration in vitro.
细胞培养：与实施例68相同。Cell culture: Same as Example 68.
实验主要试剂：Transwell小室(美国康宁公司)。Main reagents in the experiment: Transwell chamber (Corning, USA).
实验步骤：在Transwell下室加入680μL含血清培养液，上室加入2*104个细胞，将300μL含I-9a19的无血清培养液重悬细胞，作为实验组。将300μL含0.1％DMSO的无血清培养液重悬细胞，作为对照组，孵育24h。随后，将小室取出，用棉棒将上室擦洗干净，放入多聚甲醛固定30min。然后放入结晶紫染色30min。清洗干净后将小室晾干。在显微镜下随机取3-5个视野拍照并计数。Experimental steps: Add 680 μL of serum-containing culture medium to the lower chamber of the Transwell, add 2*10 4 cells to the upper chamber, and resuspend the cells in 300 μL of serum-free culture medium containing I-9a19 to serve as the experimental group. Resuspend the cells in 300 μL of serum-free culture medium containing 0.1% DMSO as a control group and incubate for 24 h. Subsequently, the small chamber was taken out, the upper chamber was scrubbed clean with a cotton swab, and paraformaldehyde was put in to fix it for 30 minutes. Then put in crystal violet staining for 30 minutes. After cleaning, dry the chamber. Take photos of 3-5 fields randomly under the microscope and count them.
实施例72 Example 72
本发明所述的化合物I-9a19体外抑制肿瘤细胞侵袭能力实验。Experiment on the ability of compound I-9a19 to inhibit tumor cell invasion in vitro.
细胞培养：与实施例68相同。Cell culture: Same as Example 68.
实验主要试剂：与实施例71相同。Main reagents in the experiment: the same as in Example 71.
实验步骤：除将Transwell上室预铺Matrigel，孵育24h更改为36h以外，其余操作与实施例71相同。Experimental steps: The rest of the operations were the same as in Example 71, except that the upper chamber of the Transwell was pre-plated with Matrigel and the incubation was changed from 24h to 36h.
实施例73Example 73
本发明所述的化合物I-9a19体内抗TNBC药效学研究实验。Research experiments on the in vivo anti-TNBC pharmacodynamics of compound I-9a19 according to the present invention.
细胞培养：与实施例68相同。Cell culture: Same as Example 68.
实验主要试剂：Matrigel基质胶(美国BD公司)、RIPA裂解液(美国Thermo Fisher Scientific公司)。The main reagents in the experiment: Matrigel Matrigel (BD Company, USA), RIPA lysis buffer (Thermo Fisher Scientific Company, USA).
实验步骤：将1×106个细胞注射至雌性BALB/c裸鼠第四对乳房垫，待肿瘤体积达到约100mm3后，将裸鼠随机分为空白溶剂组、对照药物多烯紫杉醇(DTX)组和化合物I-9a19处理组。开始第1天给药，连续给药19天后，处死裸鼠，获得肿瘤组织，称重，计算得到TGI。然后取10mg肿瘤，加入RIPA裂解液，随后离心，并收集上清，使用BCA法进行蛋白定量，按照免疫印迹实验进行后续操作。Experimental steps: Inject 1×10 6 cells into the fourth pair of breast pads of female BALB/c nude mice. After the tumor volume reaches approximately 100 mm 3 , the nude mice are randomly divided into blank solvent group and control drug docetaxel (DTX). ) group and compound I-9a19 treated group. Administration was started on the first day, and after 19 consecutive days of administration, the nude mice were sacrificed, tumor tissues were obtained, weighed, and TGI was calculated. Then take 10 mg of tumor, add RIPA lysis buffer, and then centrifuge, collect the supernatant, use the BCA method for protein quantification, and perform subsequent operations according to the Western blot experiment.
综上，基于表面等离子共振的结合活性测试表明，式(I)所示的1，4-苯二氮类化合物具有亚微摩尔级的ANXA3结合活性，活性结果如表1所示。基于蛋白免疫印迹测试实验表明，所述化合物能够在TNBC细胞中诱导ANXA3蛋白降解，活性具有微摩尔级，结果如表1所示。其中代表性异构体化合物I-9a19、(R)-I-9a19或(S)-I-9a19的降解活性结果如图1所示。In summary, the binding activity test based on surface plasmon resonance shows that the 1,4-benzenediazepine represented by formula (I) The compound has submicromolar ANXA3 binding activity, and the activity results are shown in Table 1. Experiments based on Western blot testing showed that the compound can induce ANXA3 protein degradation in TNBC cells, with activity at the micromolar level, and the results are shown in Table 1. The degradation activity results of representative isomer compounds I-9a19, (R)-I-9a19 or (S)-I-9a19 are shown in Figure 1.
通过多种肿瘤细胞的体外抗肿瘤活性测试表明，所述化合物具有微摩尔级的抗肿瘤活性，活性结果如表1所示。尤其是化合物I-9a19能够明显抑制TNBC细胞的克隆、迁移和侵袭等功能，活性结果如图2所示。尤其是化合物I-9a19在体内TNBC移植瘤模型中，显示出有效的治疗效果和诱导肿瘤组织ANXA3蛋白降解作用，活性结果如图3所示。In vitro anti-tumor activity tests on various tumor cells show that the compound has anti-tumor activity at the micromolar level, and the activity results are shown in Table 1. In particular, compound I-9a19 can significantly inhibit the cloning, migration and invasion of TNBC cells. The activity results are shown in Figure 2. In particular, compound I-9a19 showed effective therapeutic effects and induced the degradation of ANXA3 protein in tumor tissue in the in vivo TNBC transplanted tumor model. The activity results are shown in Figure 3.
表1化合物式I对ANXA3的亲和力、降解活性及其对不同肿瘤细胞株的抑制活性


Table 1 Affinity, degradation activity of compound formula I towards ANXA3 and its inhibitory activity against different tumor cell lines


亲和力KD、降解活性DC50和半数抑制浓度IC50：*：＞100μM,**：10-100μM,***：1-10μM,****：＜1μM。 Affinity K D , degradation activity DC 50 and half inhibitory concentration IC 50 : *: >100 μM, **: 10-100 μM, ***: 1-10 μM, ****: <1 μM.

Claims (9)

  1. 式(I)所示1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体，
    1,4-Benzodiazepine shown in formula (I) compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof,
    式(I)中，In formula (I),
    R1选自氢原子或甲基；R 1 is selected from a hydrogen atom or a methyl group;
    X是链接基团，选自： 其中，R2选自：X is a linking group, selected from: Where, R 2 is selected from:
    1)取代苯，其结构为：1) Substituted benzene, its structure is:
    其中， in,
    R3-R7分别独立地选自氢、卤素、甲氧基、三氟甲氧基、三氟甲基、甲基、乙基、二甲氨基、硝基、氰基或乙酰基；R 3 to R 7 are each independently selected from hydrogen, halogen, methoxy, trifluoromethoxy, trifluoromethyl, methyl, ethyl, dimethylamino, nitro, cyano or acetyl;
    2)芳香六元杂环，其结构为：2) Aromatic six-membered heterocycle, its structure is:
    其中， in,
    R8-R11分别独立地选自氢、卤素、氰基、硝基、三氟甲基或甲氧基；R 8 to R 11 are each independently selected from hydrogen, halogen, cyano, nitro, trifluoromethyl or methoxy;
    R’8-R’11分别独立地选自氢、卤素、氰基、硝基、三氟甲基或甲氧基；R' 8 -R' 11 are each independently selected from hydrogen, halogen, cyano, nitro, trifluoromethyl or methoxy;
    R12-R14分别独立地选自氢或卤素；R 12 to R 14 are each independently selected from hydrogen or halogen;
    R’12-R’14分别独立地选自氢或三氟甲基；R' 12 to R' 14 are each independently selected from hydrogen or trifluoromethyl;
    R”12-R”14分别独立地选自氢；R” 12 to R” 14 are each independently selected from hydrogen;
    3)芳香五元杂环，其结构为：3) Aromatic five-membered heterocycle, its structure is:
    其中， in,
    A选自氮、氧或硫；A is selected from nitrogen, oxygen or sulfur;
    B选自碳或氮； B is selected from carbon or nitrogen;
    C选自碳或氮；C is selected from carbon or nitrogen;
    D选自碳或氮；D is selected from carbon or nitrogen;
    E选自氮或氧；E is selected from nitrogen or oxygen;
    F选自碳或氮；F is selected from carbon or nitrogen;
    R15-R18分别独立地选自氢、甲基或三氟甲基；R 15 to R 18 are each independently selected from hydrogen, methyl or trifluoromethyl;
    R’15-R’17分别独立地选自氢或甲基；R' 15 to R' 17 are each independently selected from hydrogen or methyl;
    4)脂肪杂环或金刚烷，其结构为：4) Aliphatic heterocycle or adamantane, its structure is:
    其中， in,
    G选自碳或氮；G is selected from carbon or nitrogen;
    R19选自甲氧基或缩乙二醇。R 19 is selected from methoxy or ethylene glycol.
  2. 根据权利要求1所述式(I)所示1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体，其特征在于，1,4-benzodiazepine shown in formula (I) according to claim 1 compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, characterized in that,
    式(I)所示的1，4-苯二氮类化合物，具体选自如下所示的化合物：



    1,4-Benzodiazepine represented by formula (I) Compounds, specifically selected from the following compounds:



  3. 根据权利要求1或2所述式(I)所示1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体，其特征在于，1,4-benzodiazepine represented by formula (I) according to claim 1 or 2 compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, characterized in that,
    选择为立体异构体时，为C3位立体构型中的R或S构型，具体选自(R)-I-9a19或(S)-I-9a19化合物：
    When it is selected as a stereoisomer, it is the R or S configuration in the C3 position configuration, specifically selected from (R)-I-9a19 or (S)-I-9a19 compounds:
  4. 权利要求1或2或3所述式(I)所示1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体与医学上可接受的载体组成的药物组合物。1,4-benzodiazepine represented by formula (I) according to claim 1 or 2 or 3 A pharmaceutical composition consisting of a compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof and a medically acceptable carrier.
  5. 权利要求1或2或3所述式(I)所示1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体，或权利要求1或2或3所述式(I)所示1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体与医学上可接受的载体组成的药物组合物在制备ANXA3降解剂中的用途。1,4-benzodiazepine represented by formula (I) according to claim 1 or 2 or 3 Compounds, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, or 1,4-benzodiazepine represented by formula (I) according to claim 1 or 2 or 3 The use of a pharmaceutical composition consisting of a compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof and a medically acceptable carrier in the preparation of an ANXA3 degrading agent.
  6. 权利要求1或2或3所述式(I)所示1，4-苯二氮类化合物、或其药学上可接 受的盐、或其立体异构体，或权利要求1或2或3所述式(I)所示1，4-苯二氮类化合物、或其药学上可接受的盐、或其立体异构体与医学上可接受的载体组成的药物组合物在制备抗肿瘤药物中的用途。1,4-benzodiazepine represented by formula (I) according to claim 1 or 2 or 3 compounds, or pharmaceutically acceptable Accepted salts, or stereoisomers thereof, or 1,4-benzodiazepine represented by formula (I) according to claim 1 or 2 or 3 The use of a pharmaceutical composition consisting of a compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof and a medically acceptable carrier in the preparation of anti-tumor drugs.
  7. 根据权利要求6所述用途，其特征在于，所述抗肿瘤药物为预防和/或治疗癌症的药物。The use according to claim 6, wherein the anti-tumor drug is a drug for preventing and/or treating cancer.
  8. 根据权利要求7所述用途，其特征在于，所述的癌症选自乳腺癌、宫颈癌、卵巢癌、肠癌、胃癌、胰腺癌、肺癌、食管癌、肝癌、白血病和黑色素瘤。The use according to claim 7, characterized in that the cancer is selected from the group consisting of breast cancer, cervical cancer, ovarian cancer, intestinal cancer, gastric cancer, pancreatic cancer, lung cancer, esophageal cancer, liver cancer, leukemia and melanoma.
  9. 根据权利要求8所述用途，其特征在于，所述抗肿瘤药物为抗三阴性乳腺癌的药物。 The use according to claim 8, wherein the anti-tumor drug is an anti-triple-negative breast cancer drug.
PCT/CN2023/112421 2022-06-15 2023-08-11 1,4-benzodiazepine compound and use thereof in preparation of anti-tumor drug WO2023241738A2 (en)

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