WO2022022354A1 - 聚乙二醇偶联药物增效剂、其制备方法及用途 - Google Patents

聚乙二醇偶联药物增效剂、其制备方法及用途 Download PDF

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WO2022022354A1
WO2022022354A1 PCT/CN2021/107555 CN2021107555W WO2022022354A1 WO 2022022354 A1 WO2022022354 A1 WO 2022022354A1 CN 2021107555 W CN2021107555 W CN 2021107555W WO 2022022354 A1 WO2022022354 A1 WO 2022022354A1
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cancer
reaction
independently
polyethylene glycol
mmol
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PCT/CN2021/107555
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English (en)
French (fr)
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李高全
刘念
曾下凡
彭永陈
梅刚
杨帅
高扬
官盛
尹仪枫
陈惠渝
娄杰
钱昆
阳享伟
刘力纬
李振威
胡凯雄
刘华
张倩
李大军
卫玉松
丁小玲
刘曦
黄力群
涂涛
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重庆阿普格雷生物科技有限公司
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Priority to US18/016,897 priority Critical patent/US20230348671A1/en
Priority to EP21850002.3A priority patent/EP4190360A1/en
Priority to JP2023504146A priority patent/JP2023537248A/ja
Publication of WO2022022354A1 publication Critical patent/WO2022022354A1/zh

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    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • 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
    • A61K31/74—Synthetic polymeric materials
    • A61K31/765—Polymers containing oxygen
    • A61K31/77—Polymers containing oxygen of oxiranes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/595—Polyamides, e.g. nylon
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/331—Polymers modified by chemical after-treatment with organic compounds containing oxygen
    • C08G65/332—Polymers modified by chemical after-treatment with organic compounds containing oxygen containing carboxyl groups, or halides, or esters thereof
    • C08G65/3322—Polymers modified by chemical after-treatment with organic compounds containing oxygen containing carboxyl groups, or halides, or esters thereof acyclic
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/333—Polymers modified by chemical after-treatment with organic compounds containing nitrogen
    • C08G65/33396—Polymers modified by chemical after-treatment with organic compounds containing nitrogen having oxygen in addition to nitrogen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the invention belongs to the technical field of medicine, and particularly relates to a polyethylene glycol conjugated drug synergist, a preparation method and application thereof.
  • Polymer conjugated drugs can greatly increase the water solubility of drug molecules; most small-molecule drugs can only be maintained in the blood circulation for a few minutes, while polymer conjugated drugs can be maintained for tens, hundreds of hours or even longer.
  • the half-life increases the bioavailability of the drug.
  • anticancer drugs are highly enriched in cancerous organs, tissues or cells, which greatly reduces the toxic and side effects caused by the full body of small molecule anticancer drugs; It is limited to the endocytosis pathway, which is conducive to the delivery of drugs to the lysosome, thereby avoiding the drug resistance caused by the pumping of p-glycoprotein; stimulating or restoring immune function is conducive to killing cancer cells.
  • Polyethylene glycol is the most successful carrier in the field of polymer conjugated drug nanomedicine, and is called the "gold standard" carrier.
  • gold standard carrier for PEGylated drug technology.
  • PEGylated drugs approved by the US FDA to enter the market
  • 1 PEGylated drug has been approved by the Chinese NMPA for marketing
  • there are also Nearly 40 new clinical drugs are undergoing Phase I, Phase II, Phase III clinical trials and entering the NDA stage, half of which are PEGylated small molecule drugs.
  • the inventors discovered that the recently developed internalizing RGD polypeptide (iRGD, CRGDK/RGPD/EC), a tumor-targeting and cell-penetrating cyclic peptide, can be activated in a neuropeptide-1-dependent, tumor-specific manner Increase the ability of anticancer drugs to enter extravascular tumor tissue, thereby enhancing the efficacy of combined anticancer drugs.
  • iRGD is difficult to function clinically due to its easy hydrolysis and short half-life.
  • iRGD-regulated polymer-conjugated drugs can significantly improve the efficacy of tumor treatment by enhancing the enrichment and penetration of drugs at tumor sites. Effect.
  • the inventor provides a polyethylene glycol-coupled drug synergist.
  • the inventor connects iRGD on a polyethylene glycol polymer carrier through a special chemical synthesis method to synthesize polyethylene glycol-coupled iRGD. It is transported to the tumor site through the blood circulation. When used in combination with anticancer drugs, iRGD can be degraded in the tumor microenvironment to enhance the efficacy of anticancer drugs.
  • the present invention provides a polyethylene glycol conjugated drug synergist represented by formula (I) or a pharmaceutically acceptable salt thereof,
  • PEG 1 is a single-arm polyethylene glycol segment, PEG 1 is connected to L 1 through a carbonyl group, or the end of PEG 1 has an amino group or an activated amino group, and the number average molecular weight of PEG 1 is 5k-40k, preferably 5k-10k or 10k-40k, more preferably 10k;
  • L1 is Each r 1 is independently 1, 2, 3, 4, 5 or 6, preferably 1, 2, 3 or 4, more preferably 3 or 4; r 2 is 1, 2, 3, 4, 5 or 6, preferably 1, 2, 3 or 4, more preferably 1 or 2;
  • Y1 and Y0 are independently selected from r 1 is 1, 2, 3, 4, 5 or 6, r 1 is preferably 1, 2, 3 or 4, r 1 is more preferably 3 or 4, and each r 2 is independently 1, 2, 3, 4, 5 or 6, each r2 is independently preferably 1, 2, 3 or 4, and each r2 is independently more preferably 1 or 2;
  • L V is selected from Each r0 is independently 1, 2, 3, 4, 5 or 6, each r0 is independently preferably 3, 4, 5 or 6, each r0 is independently more preferably 5 or 6, and each r2 is independently is 1, 2, 3, 4, 5 or 6, each r2 is independently preferably 1, 2, 3 or 4, and each r2 is independently more preferably 1 or 2;
  • L 1 is N
  • Y1, Y0 are each independently selected from
  • Y1 is selected from r 1 is 3 or 4, and each r 2 is independently 1 or 2.
  • Y1 is selected from
  • Y0 is selected from r 1 is 3 or 4, and r 2 is 1 or 2.
  • Y0 is selected from
  • LV is selected from
  • the present invention provides a polyethylene glycol conjugated drug synergist represented by formula (I') or a pharmaceutically acceptable salt thereof,
  • PEG 1 is a single-arm polyethylene glycol segment, the end of PEG 1 has an amino group or an activated amino group, and the number-average molecular weight of PEG 1 is 5k-40k, preferably 5k-10k or 10k-40k, more preferably 10k;
  • L1 is r 1 is 1, 2, 3, 4, 5 or 6, preferably 1, 2, 3 or 4, more preferably 3 or 4;
  • Y1 and Y0 are independently selected from r 1 is 1, 2, 3, 4, 5 or 6, r 1 is preferably 1, 2, 3 or 4, r 1 is more preferably 3 or 4, and each r 2 is independently 1, 2, 3, 4, 5 or 6, each r2 is independently preferably 1, 2, 3 or 4, and each r2 is independently more preferably 1 or 2;
  • L V is selected from Each r0 is independently 1, 2, 3, 4, 5 or 6, each r0 is independently preferably 3, 4, 5 or 6, each r0 is independently more preferably 5 or 6, and each r2 is independently is 1, 2, 3, 4, 5 or 6, each r2 is independently preferably 1, 2, 3 or 4, and each r2 is independently more preferably 1 or 2;
  • L 1 is N
  • Y1, Y0 are each independently selected from
  • Y1 is selected from r 1 is 3 or 4, and each r 2 is independently 1 or 2.
  • Y1 is selected from
  • Y0 is selected from r 1 is 3 or 4, and r 2 is 1 or 2.
  • Y0 is selected from
  • LV is selected from
  • the present invention provides a polyethylene glycol conjugated drug synergist or a pharmaceutically acceptable salt thereof, wherein the polyethylene glycol conjugated drug synergist is selected from:
  • the present invention provides a method for preparing the aforementioned polyethylene glycol conjugated drug synergist or a pharmaceutically acceptable salt thereof, comprising:
  • M 1 , L 1 , Y 1 , Y 0 are as described above,
  • Pro 2 is a protecting group for carboxyl, preferably, Pro 2 is benzyloxy,
  • Pro 1 is the protecting group of amino or carboxyl, preferably, when Pro 1 is the protecting group of amino, Pro 1 is tert-butoxycarbonyl, preferably, when Pro 1 is the protecting group of carboxyl, Pro 1 is tert-butoxy;
  • the intermediate Choose from the following:
  • the present invention provides an intermediate for preparing the aforementioned polyethylene glycol conjugated drug synergist or a pharmaceutically acceptable salt thereof, and the intermediate is selected from the following:
  • the present invention provides a composition comprising the aforementioned polyethylene glycol conjugated drug synergist or a pharmaceutically acceptable salt thereof.
  • the composition further comprises one or more pharmaceutically acceptable adjuvants, such as carriers and/or excipients.
  • the carriers and/or excipients include, but are not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum albumin such as human serum albumin, buffer substances such as phosphate, glycerol, sorbic acid, potassium sorbate , Partial glyceride mixture of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyethylene Pyrrolidone, cellulosic material, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, beeswax, polyethylene-polyoxypropylene block polymer and lanolin.
  • the composition can be formulated into any pharmaceutically acceptable dosage form.
  • the composition may also be administered to an individual in need of such treatment by any suitable mode of administration, eg, oral, parenteral, rectal or pulmonary administration.
  • the composition can be made into conventional solid preparations, such as tablets, capsules, pills, granules, etc.; it can also be made into oral liquid preparations, such as oral solution, oral suspension, Syrup, etc.
  • suitable fillers, binders, disintegrants, lubricants and the like can be added.
  • parenteral administration the composition can be prepared into injections, including injection solutions, sterile powders for injection and concentrated solutions for injection.
  • the injection When the injection is prepared, it can be produced by the conventional method in the existing pharmaceutical field. When the injection is prepared, no additive can be added, or an appropriate additive can be added according to the properties of the drug.
  • the compositions For rectal administration, the compositions may be formulated as suppositories and the like.
  • the composition For pulmonary administration, the composition may be formulated as an inhalation or spray or the like.
  • the composition of the present invention can be formulated into injections, such as injection solutions.
  • physiological saline is used as the carrier of the injection.
  • the composition further comprises an anticancer drug.
  • the anticancer drug is a polyethylene glycol conjugated drug represented by formula (A) or a pharmaceutically acceptable salt thereof,
  • j 3 or 4;
  • PEG 2 is a single-arm polyethylene glycol segment, PEG 2 is connected to L 2 through a carbonyl group, or the end of PEG 2 has an amino group or an activated amino group, and the number-average molecular weight of PEG 2 is 5k-40k, preferably 5k-10k or 10k-40k, more preferably 5k;
  • each r 1 is independently 1, 2, 3, 4, 5 or 6, each r 1 is independently preferably 1, 2, 3 or 4, and each r 1 is independently more preferably 3 or 4;
  • Z 2 , Z 1 , and Z 0 are each independently r 1 is 1, 2, 3, 4, 5 or 6, r 1 is preferably 1, 2, 3 or 4, r 1 is more preferably 3 or 4, and each r 2 is independently 1, 2, 3, 4, 5 or 6, each r2 is independently preferably 1, 2, 3 or 4, each r2 is independently more preferably 1 or 2,
  • N is or G
  • r 0 is 1, 2, 3, 4, 5 or 6, preferably 3, 4, 5 or 6, more preferably 5 or 6;
  • AC is SB7, NPB, SN38, LPT, PCB, DOX, PTX or AXT, preferably PTX or SN38.
  • L is N
  • Z 2 , Z 1 , Z 0 are each independently
  • Z is r 1 is 3 or 4.
  • Z is
  • Z 1 is Each r 2 is independently 1 or 2.
  • Z 1 is
  • Z 0 is r 2 is 1 or 2.
  • Z 0 is
  • N is or G.
  • the polyethylene glycol conjugated drug is selected from:
  • the present invention provides the use of the aforementioned polyethylene glycol conjugated drug synergist or a pharmaceutically acceptable salt thereof or the aforementioned composition in the preparation of a medicament for enhancing Efficacy in the treatment and/or prevention of disease.
  • the disease is cancer selected from colon cancer, leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblastoma, glioma, breast cancer, adenoma/ Carcinoid, adrenocortical carcinoma, pancreatic islet cell carcinoma, cervical cancer, endometrial cancer, ovarian cancer, colorectal cancer, skin cancer, esophageal cancer, eye cancer, gallbladder cancer, stomach cancer, head and neck cancer, liver cancer, melanoma, card Posterior sarcoma, kidney cancer, oral cancer, lung cancer, nasopharyngeal cancer, neuroblastoma, ovarian cancer, pancreatic cancer, thyroid cancer, parathyroid penile cancer, prostate cancer, urethral cancer, vaginal cancer, vulvar cancer, anal cancer , sarcoma, and metastasis of the cancer.
  • colon cancer leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblast
  • the present invention provides a method for enhancing the therapeutic effect of treating and/or preventing a disease, comprising administering to an individual in need thereof an effective amount of the aforementioned polyethylene glycol conjugated drug synergist or its A pharmaceutically acceptable salt or a composition of the foregoing.
  • the disease is cancer selected from colon cancer, leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblastoma, glioma, breast cancer, adenoma/ Carcinoid, adrenocortical carcinoma, pancreatic islet cell carcinoma, cervical cancer, endometrial cancer, ovarian cancer, colorectal cancer, skin cancer, esophageal cancer, eye cancer, gallbladder cancer, stomach cancer, head and neck cancer, liver cancer, melanoma, card Posterior sarcoma, kidney cancer, oral cancer, lung cancer, nasopharyngeal cancer, neuroblastoma, ovarian cancer, pancreatic cancer, thyroid cancer, parathyroid penile cancer, prostate cancer, urethral cancer, vaginal cancer, vulvar cancer, anal cancer , sarcoma, and metastasis of the cancer.
  • colon cancer leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblast
  • the present invention provides the aforementioned polyethylene glycol conjugated drug synergist or a pharmaceutically acceptable salt thereof or the aforementioned composition for enhancing the curative effect of treating and/or preventing diseases .
  • the disease is cancer selected from colon cancer, leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblastoma, glioma, breast cancer, adenoma/ Carcinoid, adrenocortical carcinoma, pancreatic islet cell carcinoma, cervical cancer, endometrial cancer, ovarian cancer, colorectal cancer, skin cancer, esophageal cancer, eye cancer, gallbladder cancer, stomach cancer, head and neck cancer, liver cancer, melanoma, card Posterior sarcoma, kidney cancer, oral cancer, lung cancer, nasopharyngeal cancer, neuroblastoma, ovarian cancer, pancreatic cancer, thyroid cancer, parathyroid penile cancer, prostate cancer, urethral cancer, vaginal cancer, vulvar cancer, anal cancer , sarcoma, and metastasis of the cancer.
  • colon cancer leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblast
  • the present invention provides the use of the aforementioned composition (comprising an anticancer drug) in the preparation of a drug for treating and/or preventing a disease that is treated by the aforementioned anticancer drug.
  • the disease is cancer selected from colon cancer, leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblastoma, glioma, breast cancer, adenoma/ Carcinoid, adrenocortical carcinoma, pancreatic islet cell carcinoma, cervical cancer, endometrial cancer, ovarian cancer, colorectal cancer, skin cancer, esophageal cancer, eye cancer, gallbladder cancer, stomach cancer, head and neck cancer, liver cancer, melanoma, card Posterior sarcoma, kidney cancer, oral cancer, lung cancer, nasopharyngeal cancer, neuroblastoma, ovarian cancer, pancreatic cancer, thyroid cancer, parathyroid penile cancer, prostate cancer, urethral cancer, vaginal cancer, vulvar cancer, anal cancer , sarcoma, and metastasis of the cancer.
  • colon cancer leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblast
  • the present invention provides a method of treating and/or preventing a disease, which comprises administering to an individual in need thereof an effective amount of the aforementioned composition (comprising an anticancer drug), the disease being the aforementioned disease treated by anticancer drugs.
  • Dosage regimens can be adjusted to provide the optimal desired response. For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is noted that dosage values may vary with the type and severity of the condition to be alleviated, and may include single or multiple doses. It is further understood that for any particular individual, the specific dosing regimen should be adjusted over time according to the needs of the individual and the professional judgment of the person administering or supervising the administration of the composition.
  • the disease is cancer selected from colon cancer, leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblastoma, glioma, breast cancer, adenoma/ Carcinoid, adrenocortical carcinoma, pancreatic islet cell carcinoma, cervical cancer, endometrial cancer, ovarian cancer, colorectal cancer, skin cancer, esophageal cancer, eye cancer, gallbladder cancer, stomach cancer, head and neck cancer, liver cancer, melanoma, card Posterior sarcoma, kidney cancer, oral cancer, lung cancer, nasopharyngeal cancer, neuroblastoma, ovarian cancer, pancreatic cancer, thyroid cancer, parathyroid penile cancer, prostate cancer, urethral cancer, vaginal cancer, vulvar cancer, anal cancer , sarcoma, and metastasis of the cancer.
  • colon cancer leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblast
  • the present invention provides the aforementioned composition (comprising an anticancer drug) for treating and/or preventing a disease, which is a disease treated by the aforementioned anticancer drug.
  • the disease is cancer selected from colon cancer, leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblastoma, glioma, breast cancer, adenoma/ Carcinoid, adrenocortical carcinoma, pancreatic islet cell carcinoma, cervical cancer, endometrial cancer, ovarian cancer, colorectal cancer, skin cancer, esophageal cancer, eye cancer, gallbladder cancer, stomach cancer, head and neck cancer, liver cancer, melanoma, card Posterior sarcoma, kidney cancer, oral cancer, lung cancer, nasopharyngeal cancer, neuroblastoma, ovarian cancer, pancreatic cancer, thyroid cancer, parathyroid penile cancer, prostate cancer, urethral cancer, vaginal cancer, vulvar cancer, anal cancer , sarcoma, and metastasis of the cancer.
  • colon cancer leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblast
  • cancer refers to a cell proliferative disease state, including but not limited to: colon cancer, leukemia, lymphoma, bladder cancer, bone cancer, brain tumor, medulloblastoma, glioma, breast cancer, adenoma/ Carcinoid, adrenocortical carcinoma, pancreatic islet cell carcinoma, cervical cancer, endometrial cancer, ovarian cancer, colorectal cancer, skin cancer, esophageal cancer, eye cancer, gallbladder cancer, stomach cancer, head and neck cancer, liver cancer, melanoma, card Posterior sarcoma, kidney cancer, oral cancer, lung cancer, nasopharyngeal cancer, neuroblastoma, ovarian cancer, pancreatic cancer, thyroid cancer, parathyroid penile cancer, prostate cancer, urethral cancer, vaginal cancer, vulvar cancer, anal cancer , sarcoma, etc., including metastasis of the aforementioned cancers.
  • colon cancer colon cancer
  • “individual” includes humans or non-human animals.
  • Exemplary human subjects include human subjects (referred to as patients) or normal subjects with a disease (eg, a disease described herein).
  • non-human animals includes all vertebrates, such as non-mammals (eg birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and/or domesticated animals (eg sheep, dogs, cats, cows, pigs, etc.).
  • the term "effective amount” refers to an amount of a compound that, when administered, will alleviate to some extent one or more symptoms of the condition being treated.
  • treating means reversing, alleviating, inhibiting the progression of a disorder or condition to which such term applies or one or more symptoms of such disorder or condition, or preventing such disorder or condition or such one or more symptoms of a disorder or condition.
  • the polyethylene glycol conjugated drug synergist of the present invention uses amino acids or polypeptides as the linking chain, and uses dicarboxylic acids or polycarboxylic acids with amino groups (such as natural amino acids with two carboxyl groups) or with dicarboxylic acid groups.
  • Amino or polyamino carboxylic acids (eg, natural amino acids with two amino groups) or polycarboxylic acids are used as bridges to couple together multiple PPT-iRGDs or MI-AH-PPT-iRGDs by forming amide bonds.
  • the activated PEG reacts with an amino group on the backbone through a carboxyl group to form an amide bond.
  • PEG reacts with a carboxyl group on the backbone through a terminal amino group to form an amide bond.
  • the molecular weight of PEG comprises its terminal amino groups (ie, PEG derivatives with reactive groups).
  • the polyethylene glycol conjugated drugs of the present invention use amino acids or polypeptides as the connecting chain, and dicarboxylic acids or polycarboxylic acids with amino groups (for example, natural amino acids with two carboxyl groups) or with diamino or polycarboxylic acids.
  • Amino carboxylic acids (eg, natural amino acids with two amino groups) or polycarboxylic acids are bridges that couple together multiple identical or different drug molecules through the formation of amide bonds.
  • the types, ratios and loadings of drugs can be adjusted.
  • the activated PEG reacts with an amino group on the backbone through a carboxyl group to form an amide bond.
  • the molecular weight of PEG comprises its terminal amino groups (ie, PEG derivatives with reactive groups).
  • the active ingredient suitable for coupling with polyethylene glycol can be a drug molecule with at least one amino group, hydroxyl group, carboxyl group or acyl group, for example, with at least one amino group, hydroxyl group, Carboxyl or acyl-containing drug molecules with anti-tumor activity, such as SB7, NPB, SN38, LPT, PCB, DOX, PTX or AXT, have the following meanings:
  • PPT-iRGD The structural formula of PPT-iRGD is The connection site of PPT-iRGD and other parts of the overall structure of the polyethylene glycol conjugated drug synergist is the terminal sulfhydryl group which is location shown.
  • the structural formula of MI-AH-PPT-iRGD is The linking site of MI-AH-PPT-iRGD and other parts of the overall structure of the polyethylene glycol conjugated drug synergist is a maleimide group which is location shown.
  • MI-AH-PPT-iRGD is addition-linked to the maleimide group with an additional sulfhydryl group.
  • connection sites of the following drug molecules and other parts of the overall structure of the polyethylene glycol conjugated drug are shown in the table below, specifically: location shown.
  • PEG polyethylene glycol
  • n in the subscript of the polyethylene glycol repeating unit represents the degree of polymerization of polyethylene glycol.
  • each independently is/selected from and “each independently are/selected from” used throughout this document are interchangeable, both It should be understood in a broad sense. It can either mean that in different groups, the specific options expressed between the same symbols do not affect each other, or it can mean that in the same group, the specific options expressed between the same symbols do not affect each other.
  • pharmaceutically acceptable salts of the compounds of the present invention include acid and base addition salts of the compounds.
  • acid and base addition salts for example, hydrochloride, hexafluorophosphate, meglumine and the like.
  • the wavy line in the structural formula means the position where other groups are bonded to the structure represented by the formula.
  • FIG. 1 is a schematic diagram showing the growth trend of each group of tumors according to the embodiment of the present invention.
  • FIG. 2 shows a schematic diagram of the tumor weight inhibition rate of each group in the examples of the present invention.
  • the sources and structures of some of the raw materials are as follows:
  • Boc-L-Lys(Fmoc)-OH (20 g, 42.6857 mmol, purchased from Aladdin), Gly-OBn (14.4 g, 42.6857 mmol, purchased from innochem), HBTU (24 g, 64.0286 mmol), HOBT (8.65 g, 64.0286 mmol) was put into a 500 mL reaction flask, DMF (40 mL) was added to dissolve, and the mixture was stirred at -5°C for 30 minutes. DIEA (31.7 mL, 192.085 mmol) was slowly added dropwise, and the reaction was carried out at -5°C overnight. After the reaction was completed, the reaction was extracted with pure water and ethyl acetate, washed with saturated brine, the organic phase was concentrated, and evaporated to dryness to obtain 26 g of the product.
  • Boc-L-Lys(Boc)-OH (7g, 21.334mmol, purchased from Aladdin), 45-115 (11g, 21.334mmol), HBTU (12.1363g, 32.0016mmol), HOBT (4.3241g, 32.0016mmol) ) into a 500 mL reaction flask, add DMF (80 mL) to dissolve, and stir for 30 minutes at -5°C.
  • DIEA (15 mL, 96.003 mmol) was slowly added dropwise, and the reaction was carried out at -5°C overnight.
  • reaction was extracted with pure water and ethyl acetate, washed with saturated brine, the organic phase was concentrated, sampled by dry method, subjected to column chromatography, and eluted with a gradient of 50%-100% ethyl acetate/petroleum ether to obtain product 15 g, 83% yield.
  • reaction solution was concentrated, dissolved in ethyl acetate (100 mL), and saturated sodium bicarbonate solution was added until the aqueous phase became alkaline, the organic phase was separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL ⁇ 3) until There was no product in the aqueous phase, the organic phases were combined, washed once with saturated brine (150 mL), concentrated and evaporated to dryness to obtain 11.44 g of the product.
  • reaction was extracted with pure water and ethyl acetate, washed with saturated brine, concentrated the organic phase, sampled by dry method, subjected to column chromatography, and eluted with a gradient of 1%-55 methanol/dichloromethane to obtain 18 g of the product, Yield 81%.
  • the reactant 45-127 (1.1 g, 0.8458 mmol) was taken and placed in a 250 mL flask, DMF solution (50 mL) was added, morpholine (1.4745 mL, 16.9155 mmol) was added, and the reaction was stirred at room temperature. After 3 hours, the reaction was completed.
  • reaction solution was extracted and treated with pure water and ethyl acetate, the organic phase was concentrated, sampled by dry method, and subjected to column chromatography.
  • the product was eluted with a gradient of 1%-2% ethyl acetate/petroleum ether to give 15 g of product in 28% yield.
  • Boc-Glu-OH (5.0g, 20.22mmol, purchased from Aladdin), H-Glu(OBzl)-OBzl ⁇ TsOH (21.2g, 42.46mmol, purchased from Ark Pharm), HOBT (8g, 60.66mmol) , HBTU (23g, 60.66mmol) was put into a 250mL reaction flask, DMF solution (80mL) was added to dissolve, the reactant was completely dissolved by ultrasound, stirred at -5°C for 30 minutes, DIEA (30mL, 181mmol) was slowly added dropwise, and the reaction was carried out at a low temperature. to the end.
  • 7-ethyl-10-hydroxycamptothecin (15.00g, 38.23mmol, abbreviated as SN38) was placed in a 1000ml round-bottomed flask, 150ml of dichloromethane was added to dissolve it, and tert-butyldiphenylchlorosilane ( 59.64 ml, 229.36 mmol, purchased from Shaoyuan), triethylamine (31.88 ml, 229.36 mmol), and the reaction was stirred at 37°C under oil bath overnight.
  • reaction solution was steamed to a viscous state, n-hexane (150 ml) was added to settle the product to a solid, suction filtration, and drying were performed to obtain the product (23.15 g, 96%).
  • Fmoc-Lys(Boc)-OH (5.0 g, 10.6714 mmol, purchased from Aladdin), H-Gly-OBn (3.7802 g, 11.2050 mmol, purchased from innochem), HBTU (6.0705 g, 16.0072 mmol), HOBT ( 2.1630g, 16.0072mmol) was added to a 500ml flask, DMF (50mL) was added to dissolve it, and the reaction was stirred at 0 degrees Celsius for 30 minutes. DIEA (7.9371 mL, 48.0215 mmol) was slowly added dropwise, and the reaction was continued to stir at 0 degrees Celsius overnight.
  • Boc-Lys(Boc)-OH (4.2805g, 11.7385mmol, purchased from Ark Pharm), 29-243 (5.50g, 10.6714mmol), HBTU (6.0705g, 16.0072mmol), HOBT (2.1630g, 16.0072mmol) It was added to a 500ml flask, DMF (50mL) was added to dissolve it, and the reaction was stirred at 0 degrees Celsius for 30 minutes. DIEA (7.9371 mL, 48.0215 mmol) was slowly added dropwise, and the reaction was continued at 0 degrees Celsius overnight.
  • Morpholine (9.24 mL, 106.714 mmol) and DMF (10 mL) were added to a 500 mL flask containing 29-245 (9.0 g, 10.6714 mmol) to dissolve and the reaction was stirred at room temperature for 1 hour.
  • the powdery solid in the reaction solution was separated out, and the filter cake was filtered with methyl tert-butyl ether (40 mL ⁇ 3 ) washing, collecting the filter cake, then dissolving with methanol/dichloromethane (1:4) solution (100 mL), adding silica gel powder (20 g), evaporating to powder, dry loading, column chromatography, using 3% -12% methanol in dichloromethane mixed solution gradient elution, collected, concentrated, and dried in a vacuum oven to obtain the product (1.74 g, 56.35%).
  • the powdery solid in the reaction solution was separated out, and the filter cake was filtered with methyl tert-butyl ether (40 mL ⁇ 3 ) washing, collecting the filter cake, then dissolving with methanol/dichloromethane (1:4) solution (100 mL), adding silica gel powder (60 g), evaporating to powder, dry loading, column chromatography, using 1% - 4% methanol in dichloromethane mixed solution gradient elution, collected and concentrated, and dried in a vacuum oven to obtain the product (2.7 g, 27.92%).
  • n-hexane 100 mL was added for shaking, the supernatant was poured out, and the above operation was repeated three times. Then methyl tert-butyl ether (80 mL) and a small amount of n-hexane (10 mL) were added for shaking, and the supernatant was poured out and repeated three times.
  • the powdery solid was separated out in the reaction solution, suction filtration, the filter cake was washed with methyl tert-butyl ether (40mL x 3), the filter cake was collected, and then dissolved with methanol/dichloromethane (1:4) solution (100mL), Silica gel powder (20 g) was added, evaporated to powder, dry loading, column chromatography, gradient elution with a mixed solution of 3%-15% methanol in dichloromethane, collected and concentrated, and dried in a vacuum oven to obtain the product (0.95 g, 37.25%).
  • reaction solution was evaporated to dryness, dissolved in DMF (5ml), and then precipitated with isopropanol, repeated three times; then dissolved in absolute ethanol and a small amount of dichloromethane, added with methyl tert-butyl ether for sedimentation, and repeated three times , the solid was collected and dried in a vacuum oven to give the product (0.75 g, 93.75%).
  • reaction was extracted with pure water and ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered with suction, concentrated in the organic phase, and evaporated to dryness to obtain 7 g of the product.
  • the reactant 38-137 (6.9 g, 8.05 mmol) was taken into a 250 mL flask, DMF solution (50 mL) was added, morpholine (14 mL, 161 mmol) was added, and the reaction was stirred at room temperature. After 3 hours, the reaction was completed. Saturated brine (150 mL) and ethyl acetate (200 mL), separated the organic phase, the aqueous phase was extracted three times with ethyl acetate (150 mL ⁇ 3), there was no product in the aqueous phase, the organic phases were combined and washed twice with saturated brine (150mL ⁇ 2), the organic phase was evaporated to dryness to obtain 5g of product.
  • the reactant 38-143 (7 g, 5.7029 mmol) was taken and placed in a 250 mL flask, DMF solution (50 mL) was added, morpholine (9..9368 mL, 114.0585 mmol) was added, and the reaction was stirred at room temperature. After 3 hours, the reaction was completed.
  • reaction solution was extracted and treated with pure water and ethyl acetate, the organic phase was concentrated, sampled by dry method, and subjected to column chromatography.
  • the product was eluted with a gradient of 1%-2% ethyl acetate/petroleum ether to obtain 5 g of the product in 20% yield.
  • reaction solution was concentrated, dissolved in ethyl acetate (100 mL), and saturated sodium bicarbonate solution was added until the aqueous phase became alkaline, the organic phase was separated, and the aqueous phase was extracted three times with ethyl acetate (50 mL ⁇ 3) until There is no product in the aqueous phase, and the organic phases are combined, washed once with saturated brine (50 mL), concentrated and evaporated to dryness to obtain 22 g of the product.
  • reaction was extracted with pure water and ethyl acetate, washed with saturated brine, the organic phase was concentrated, sampled by dry method, subjected to column chromatography, and eluted with a gradient of 50%-100% ethyl acetate/petroleum ether to obtain product 15 g, 83% yield.
  • reaction solution was concentrated, dissolved in ethyl acetate (100 mL), and saturated sodium bicarbonate solution was added until the aqueous phase became alkaline, the organic phase was separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL ⁇ 3) until There was no product in the aqueous phase, the organic phases were combined, washed once with saturated brine (150 mL), concentrated and evaporated to dryness to obtain 11.44 g of the product.
  • reaction was extracted with pure water and ethyl acetate, washed with saturated brine, concentrated the organic phase, sampled by dry method, subjected to column chromatography, and eluted with a gradient of 1%-55 methanol/dichloromethane to obtain 18 g of the product, Yield 81%.
  • the reactant 45-127 (1.1 g, 0.8458 mmol) was taken and placed in a 250 mL flask, DMF solution (50 mL) was added, morpholine (1.4745 mL, 16.9155 mmol) was added, and the reaction was stirred at room temperature. After 3 hours, the reaction was completed.
  • reaction solution was extracted and treated with pure water and ethyl acetate, the organic phase was concentrated, sampled by dry method, and subjected to column chromatography.
  • the product was eluted with a gradient of 1%-2% ethyl acetate/petroleum ether to obtain 5 g of the product in 20% yield.
  • reaction was extracted with pure water and ethyl acetate, washed with saturated brine, the organic phase was concentrated, sampled by dry method, subjected to column chromatography, and eluted with a gradient of 50%-100% ethyl acetate/petroleum ether to obtain product 15 g, 83% yield.
  • reaction solution was concentrated, dissolved in ethyl acetate (100 mL), and saturated sodium bicarbonate solution was added until the aqueous phase became alkaline, the organic phase was separated, and the aqueous phase was extracted three times with ethyl acetate (150 mL ⁇ 3) until There was no product in the aqueous phase, the organic phases were combined, washed once with saturated brine (150 mL), concentrated and evaporated to dryness to obtain 11.44 g of the product.
  • reaction was extracted with pure water and ethyl acetate, washed with saturated brine, concentrated the organic phase, sampled by dry method, subjected to column chromatography, and eluted with a gradient of 1%-55 methanol/dichloromethane to obtain 18 g of the product, Yield 81%.
  • the reactant 45-127 (1.1 g, 0.8458 mmol) was taken and placed in a 250 mL flask, DMF solution (50 mL) was added, morpholine (1.4745 mL, 16.9155 mmol) was added, and the reaction was stirred at room temperature. After 3 hours, the reaction was completed.
  • reaction solution was extracted with pure water and ethyl acetate, and the organic phase was concentrated, sampled by dry method, and subjected to column chromatography.
  • the product was eluted with a gradient of 1%-2% ethyl acetate/petroleum ether to obtain 5 g in a 20% yield.
  • Example 2 The synergistic test of the anti-tumor effect of polyethylene glycol conjugated drug synergist on the anti-cancer drug in the subcutaneous xenograft model of human breast cancer MDA-MB-231 cells BALB/c nude mice
  • Negative control use normal saline directly.
  • the prepared test article and reference substance should be stored at 2-8°C or in an ice box before administration, and the remaining test article and reference substance should be treated as medical waste.
  • Human breast cancer cell MDA-MB-231 from the Cell Resource Center, Institute of Basic Sciences, Chinese Academy of Medical Sciences, cultured in RPMI1640+10% FBS, 37°C, 5% CO 2 .
  • Age at the time of tumor inoculation about 4-5 weeks.
  • Weight at the time of tumor inoculation about 15 to 18 g.
  • the body weight of animals of the same sex ranges from 80% to 120% of the average body weight.
  • Gender and quantity female, 96 mice were ordered, 48 model animals were screened for the final experiment, and the remaining animals were transferred to a veterinarian or euthanized.
  • Animals were kept in an independent ventilation system (IVC), with a maximum of 6 animals in the same group per cage, SPF animal room, and the environmental conditions were controlled at room temperature of 20-26°C, relative humidity of 40%-70%, and alternating light and dark for 12 hours.
  • IVC independent ventilation system
  • qualified rat feed production unit: Beijing Keao Xieli Feed Co., Ltd.
  • the MDA-MB-231 cells were revived, and the cells were passaged and expanded. When a sufficient number of cells were expanded, the cells in the logarithmic growth phase were collected and prepared for cell inoculation.
  • the cell concentration at 1 ⁇ 10 8 cells/mL, and mix with Matrigel Basement Membrance Matrix (BD Company) at a volume of 1:1 to obtain a cell suspension with a concentration of 5 ⁇ 10 7 cells/mL. , 0.2mL/inoculated subcutaneously in the right armpit of 96 mice.
  • the tumor growth was observed after inoculation, and 48 tumor-forming animals with tumor volume of 77.30-292.27 mm 3 were finally screened for the experiment.
  • tumor volume and body weight they were randomly divided into 8 groups, among which group 1 (negative control group, normal saline), group 6 (49-166, 48mg/kg), group 8 (49-166+45-164, 48+176mg) /kg), 6 in each group.
  • group 1 negative control group, normal saline
  • group 6 49-166, 48mg/kg
  • group 8 49-166+45-164, 48+176mg/kg
  • each group was administered at D1, D4, D7, D10, D13, D18, D21, D24, and animals were euthanized at D27.
  • Observation content including the animal's mental state, behavioral activities, feeding situation, etc.
  • Detection time after receiving, before inoculation, grouping (that is, before the first administration) (D1), twice a week thereafter, and weighing the animals before euthanasia. Animals are also weighed in the event of accidental death or near-death euthanasia.
  • Detection time grouping (that is, the day of the first administration is D1), twice a week after the first administration, and before euthanasia, the vernier caliper measures and records the tumor length and short diameter, and calculates the tumor volume.
  • Tumor volume was calculated according to the following formula:
  • V 1/2 ⁇ long diameter ⁇ short diameter 2
  • ANOVA One-way analysis of variance
  • V t tumor volume obtained by measuring the tumor daily
  • V 0 initial tumor volume (before administration)
  • T/C% Mean RTV of the administration group/Mean RTV of the control group ⁇ 100%
  • the relative tumor proliferation rate T/C% of the administration group is less than or equal to 40% and the RTV of this group is effective compared with the RTV of the negative control group with a P less than 0.05, which has tumor growth inhibitory effect; on the contrary, if T/C%>40%, then It has no inhibitory effect on tumor growth.
  • the tumor nodules were peeled off and weighed, and the difference in tumor weight between each group was compared to further calculate the tumor inhibition rate IR TW , with IR TW ⁇ 60% as the auxiliary reference index for effectiveness, and the calculation formula was as follows:
  • IR TW (%) (W model group- W administration group )/W model group ⁇ 100%
  • Photographs were taken of euthanized animals and tumors.
  • the tumor gradually increased throughout the experiment, and at the end of the experiment (D27), the average tumor volume in group 1 was 2962.92 ⁇ 2176.59mm 3 , and the average RTV was 19.14 ⁇ 12.01; the average tumor volume in groups 6 and 8 They were 1690.27 ⁇ 785.04mm 3 and 1273.37 ⁇ 358.45mm 3 respectively, and the average RTVs were 13.03 ⁇ 5.43 and 10.55 ⁇ 5.73, respectively.
  • mice were euthanized and the tumor weights were weighed.
  • the average tumor weights in groups 1, 6, and 8 were 2.555 ⁇ 2.207g, 0.990 ⁇ 0.399g, and 0.684 ⁇ 0.165g, respectively.
  • the IR TW % of groups 6 and 8 were 61.24% and 73.21%, respectively.
  • Figure 2 shows a schematic diagram of the inhibition rate of tumor weight in each group.
  • test articles 49-166 at 48mg/kg, 49-166 at 48mg/kg and 45-164 at 176mg/kg were combined (49-166+45-164).
  • Intravenous injection has obvious inhibitory effect on the growth of human breast cancer MDA-MB-231 cells BALB/c nude mice subcutaneously transplanted tumor, and the tumor inhibitory effect of 49-166+45-164 is significantly better than that of 49-166.
  • the polyethylene glycol conjugated drug synergist of the present invention can be used as an anticancer drug on subcutaneously transplanted tumors (for example, human breast cancer MDA-MB-231 cells BALB/c nude mice subcutaneously transplanted).
  • subcutaneously transplanted tumors for example, human breast cancer MDA-MB-231 cells BALB/c nude mice subcutaneously transplanted.
  • the synergist of anti-tumor effect of tumor), and the synergistic effect is obvious.

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Abstract

一种聚乙二醇偶联药物增效剂、其制备方法及用途，具体涉及式I所示的聚乙二醇偶联药物增效剂或其药学上可接受的盐，所述聚乙二醇偶联药物增效剂或其药学上可接受的盐的制备方法，用于制备所述聚乙二醇偶联药物增效剂或其药学上可接受的盐的中间体，包含所述聚乙二醇偶联药物增效剂或其药学上可接受的盐的组合物，以及所述聚乙二醇偶联药物增效剂或其药学上可接受的盐在制备药物中的用途。

Description

聚乙二醇偶联药物增效剂、其制备方法及用途 技术领域
本发明属医药技术领域，具体涉及聚乙二醇偶联药物增效剂、其制备方法及用途。
背景技术
高分子偶联药物可以大幅度增加药物分子的水溶性；大多数的小分子药物在血液循环中只能保持数分钟，而高分子偶联药物可以保持几十、几百小时甚至更长，这有利于因肿瘤毛细血管的渗漏而产生的“增强渗透和保留”效应(即EPR效应)；由于流体力学体积增加而减弱了药物的肾消除，保护药物不被酶降解，延长药物在血浆内的半衰期，增加药物的生物利用度。此外，通过主动靶向或EPR被动靶向使抗癌药高度富集在癌变的器官、组织或细胞，大幅度地降低因为小分子抗癌药充满全身而导致的毒副作用；将药物的细胞吸收局限于内吞路径，有利于向溶酶体方向的药物传输，从而避开p-糖蛋白泵出而导致的抗药性；刺激或恢复免疫功能，利于杀灭癌细胞。
聚乙二醇是高分子偶联药物纳米医学领域最成功的载体，被称之为“黄金标准”载体。过去30年来，聚乙二醇化药物技术取得了巨大的成就，已经有17个聚乙二醇化药物被美国FDA批准进入市场销售，有1个聚乙二醇化药物被中国NMPA批准上市，此外还有近40个临床新药在进行一期、二期、三期临床试验和进入NDA阶段，其中半数为聚乙二醇化小分子药物。
发明内容
本申请是基于发明人对以下事实和问题的发现和认识作出的：
发明人发现，最近开发出来的内化RGD多肽(iRGD，CRGDK/RGPD/EC)，是一种肿瘤靶向和细胞穿透的环肽，可通过依赖于神经肽-1的、肿瘤特异的方式增加抗癌药进入血管外肿瘤组织的能力，从而增强联用抗癌药的疗效。不过，iRGD在临床上却因为很容易水解、半衰期太短很难发挥作用，而iRGD调节的高分子偶联药物却可以通过增强药物在肿瘤部位的富集和穿透性来显著提高肿瘤治疗的效果。为此，发明人提供了一种聚乙二醇偶联药物增效剂，发明人将iRGD通过特殊的化学合成方法连接在聚乙二醇高分子载体上，合成聚乙二醇偶联iRGD，其经血液循环运输到肿瘤部位，在与抗癌药联用时，iRGD在肿瘤微环境中可以被降解出来，增强抗癌药的疗效。
在本发明的第一方面，本发明提供了式(I)所示的聚乙二醇偶联药物增效剂或其药学上可接受的盐，
Figure PCTCN2021107555-appb-000001
其中：
M 1为
Figure PCTCN2021107555-appb-000002
PEG 1为单臂聚乙二醇链段，PEG 1通过羰基与L 1相连或者PEG 1的末端带有氨基或活化的氨基，PEG 1的数均分子量为5k-40k，优选为5k-10k或10k-40k，更优选为10k；
L 1为
Figure PCTCN2021107555-appb-000003
各r 1独立地为1、2、3、4、5或6，优选为1、2、3或4，更优选为3或4；r 2为1、2、3、4、5或6，优选为1、2、3或4，更优选为1或2；
V为
Figure PCTCN2021107555-appb-000004
Y1、Y0各自独立地选自
Figure PCTCN2021107555-appb-000005
Figure PCTCN2021107555-appb-000006
r 1为1、2、3、4、5或6，r 1优选为1、2、3或4，r 1更优选为3或4，各r 2独立地为1、2、3、4、5或6，各r 2独立地优选为1、2、3或4，各r 2独立地更优选为1或2；
P为-L V-T；
L V选自
Figure PCTCN2021107555-appb-000007
Figure PCTCN2021107555-appb-000008
各r 0独立地为1、2、3、4、5或6，各r 0独立地优选为3、4、5或6，各r 0独立地更优选为5或6，各r 2独立地为1、2、3、4、5或6，各r 2独立地优选为1、2、3或4，各r 2独立地更优选为1或2；
T为
Figure PCTCN2021107555-appb-000009
在一些实施方案中，L 1为
Figure PCTCN2021107555-appb-000010
在一些实施方案中，Y1、Y0各自独立地选自
Figure PCTCN2021107555-appb-000011
Figure PCTCN2021107555-appb-000012
在一些实施方案中，Y1选自
Figure PCTCN2021107555-appb-000013
r 1为3或4，各r 2独立地为1或2。
在一些实施方案中，Y1选自
Figure PCTCN2021107555-appb-000014
在一些实施方案中，Y0选自
Figure PCTCN2021107555-appb-000015
r 1为3或4，r 2为1或2。
在一些实施方案中，Y0选自
Figure PCTCN2021107555-appb-000016
在一些实施方案中，L V选自
Figure PCTCN2021107555-appb-000017
Figure PCTCN2021107555-appb-000018
在本发明的第二方面，本发明提供了式(I’)所示的聚乙二醇偶联药物增效剂或其药学上可接受的盐，
Figure PCTCN2021107555-appb-000019
其中：
M 1为
Figure PCTCN2021107555-appb-000020
PEG 1为单臂聚乙二醇链段，PEG 1的末端带有氨基或活化的氨基，PEG 1的数均分子量为5k-40k，优选为5k-10k或10k-40k，更优选为10k；
L 1为
Figure PCTCN2021107555-appb-000021
r 1为1、2、3、4、5或6，优选为1、2、3或4，更优选为3或4；
V为
Figure PCTCN2021107555-appb-000022
Y1、Y0各自独立地选自
Figure PCTCN2021107555-appb-000023
Figure PCTCN2021107555-appb-000024
r 1为1、2、3、4、5或6，r 1优选为1、2、3或4，r 1更优选为3或4，各r 2独立地为1、2、3、4、5或6，各r 2独立地优选为1、2、3或4，各r 2独立地更优选为1或2；
P为-L V-T；
L V选自
Figure PCTCN2021107555-appb-000025
各r 0独立地为1、2、3、4、5或6，各r 0独立地优选为3、4、5或6，各r 0独立地更优选为5或6，各r 2独立地为1、2、3、4、5或6，各r 2独立地优选为1、2、3或4，各r 2独立地更优选为1或2；
T为
Figure PCTCN2021107555-appb-000026
在一些实施方案中，L 1为
Figure PCTCN2021107555-appb-000027
在一些实施方案中，Y1、Y0各自独立地选自
Figure PCTCN2021107555-appb-000028
Figure PCTCN2021107555-appb-000029
在一些实施方案中，Y1选自
Figure PCTCN2021107555-appb-000030
r 1为3或4，各r 2独立地为1或2。
在一些实施方案中，Y1选自
Figure PCTCN2021107555-appb-000031
在一些实施方案中，Y0选自
Figure PCTCN2021107555-appb-000032
r 1为3或4，r 2为1或2。
在一些实施方案中，Y0选自
Figure PCTCN2021107555-appb-000033
在一些实施方案中，L V选自
Figure PCTCN2021107555-appb-000034
Figure PCTCN2021107555-appb-000035
在本发明的第三方面，本发明提供了聚乙二醇偶联药物增效剂或其药学上可接受的盐，其中，所述聚乙二醇偶联药物增效剂选自：
Figure PCTCN2021107555-appb-000036
Figure PCTCN2021107555-appb-000037
Figure PCTCN2021107555-appb-000038
Figure PCTCN2021107555-appb-000039
Figure PCTCN2021107555-appb-000040
Figure PCTCN2021107555-appb-000041
在本发明的第四方面，本发明提供了制备前述的聚乙二醇偶联药物增效剂或其药学上可接受的盐的方法，其包括：
(1)制备中间体
Figure PCTCN2021107555-appb-000042
其中：
M 1、L 1、Y 1、Y 0如前所述，
Pro 2为羧基的保护基，优选地，Pro 2为苄氧基，
Pro 1为氨基或羧基的保护基，优选地，Pro 1为氨基的保护基时，Pro 1为叔丁氧羰基，优选地，Pro 1为羧基的保护基时，Pro 1为叔丁氧基；
(2)将中间体
Figure PCTCN2021107555-appb-000043
进行第一脱保护，得到中间体
Figure PCTCN2021107555-appb-000044
其中，
Figure PCTCN2021107555-appb-000045
的末端带有羧基，；
(3)使中间体
Figure PCTCN2021107555-appb-000046
与PEG 1发生酰胺化反应，得到中间体
Figure PCTCN2021107555-appb-000047
其中，PEG 1如前所述；
(4)Pro 1为氨基的保护基时，将中间体
Figure PCTCN2021107555-appb-000048
进行第二脱保护，得到中间体
Figure PCTCN2021107555-appb-000049
其中，
Figure PCTCN2021107555-appb-000050
的末端带有氨基；
或者，Pro 1为羧基的保护基时，将中间体
Figure PCTCN2021107555-appb-000051
进行第三脱保护，得到中间体
Figure PCTCN2021107555-appb-000052
其中，
Figure PCTCN2021107555-appb-000053
的末端带有羧基；
(5)使中间体
Figure PCTCN2021107555-appb-000054
与
Figure PCTCN2021107555-appb-000055
发生酰胺化反应，得到中间体
Figure PCTCN2021107555-appb-000056
其中，r 0如前所述；
或者，使中间体
Figure PCTCN2021107555-appb-000057
与
Figure PCTCN2021107555-appb-000058
Figure PCTCN2021107555-appb-000059
发生酰胺化反应，得到中间体
Figure PCTCN2021107555-appb-000060
其中，r 0、r 2如前所述；
(6)使中间体
Figure PCTCN2021107555-appb-000061
Figure PCTCN2021107555-appb-000062
与
Figure PCTCN2021107555-appb-000063
发生加成反应，得到前述的聚乙二醇偶联药物增效剂。
在一些实施方案中，中间体
Figure PCTCN2021107555-appb-000064
选自以下：
Figure PCTCN2021107555-appb-000065
Figure PCTCN2021107555-appb-000066
Figure PCTCN2021107555-appb-000067
在本发明的第五方面，本发明提供了中间体，其用于制备前述聚乙二醇偶联药物增效剂或其药学上可接受的盐，所述中间体选自以下：
Figure PCTCN2021107555-appb-000068
Figure PCTCN2021107555-appb-000069
Figure PCTCN2021107555-appb-000070
在本发明的第六方面，本发明提供了组合物，其包含前述聚乙二醇偶联药物增效剂或其药学上可接受的盐。
在一些实施方案中，所述组合物还包含一种或多种药学上可接受的辅料，例如载体和/或赋形剂。所述载体和/或赋形剂包括但不限于:离子交换剂,氧化铝,硬脂酸铝,卵磷脂,血清蛋白如人血清蛋白,缓冲物质如磷酸盐,甘油,山梨酸,山梨酸钾,饱和植物脂肪酸的部分甘油酯混合物，水，盐或电解质，如硫酸鱼精蛋白，磷酸氢二钠，磷酸氢钾，氯化钠，锌盐，胶态氧化硅，三硅酸镁，聚乙烯吡咯烷酮，纤维素物质，聚乙二醇，羧甲基纤维素钠，聚丙烯酸酯，蜂蜡，聚乙烯-聚氧丙烯嵌段聚合物和羊毛脂。
所述组合物可以制成药学上可接受的任一剂型。所述组合物还可以以任何合适的给药方式，例如口服、肠胃外、直肠或经肺给药等方式施用于需要这种治疗的个体。用于口服给药时，所述组合物可制成常规的固体制剂，如片剂、胶囊剂、丸剂、颗粒剂等；也可制成口服液体制剂，如口服溶液剂、口服混悬剂、糖浆剂等。制成口服制剂时，可以加入适宜的填充剂、粘合剂、崩解剂、润滑剂等。用于肠胃外给药时，所述组合物可制成注射剂，包括注射液、注射用无菌粉末与注射用浓溶液。制成注射剂时，可采用现有制药领域中的常规方法生产，配制注射剂时，可以不加入附加剂，也可根据药物的性质加入适宜的附加剂。用于直肠给药时，所述组合物可制成栓剂等。用于经肺给药时，所述组合物可制成吸入剂或喷雾剂等。优选地，本发明的组合物可以被制成注射剂，例如注射液。可选地，以生理盐水作为所述注射液的载体。
在一些实施方案中，所述组合物还包含抗癌药物。
在一些实施方案中，所述抗癌药物为式(A)所示的聚乙二醇偶联药物或其药学上可接受的盐，
Figure PCTCN2021107555-appb-000071
其中：
M 2为
Figure PCTCN2021107555-appb-000072
j为3或4；
PEG 2为单臂聚乙二醇链段，PEG 2通过羰基与L 2相连或者PEG 2的末端带有氨基或活化的氨基，PEG 2的数均分子量为5k-40k，优选为5k-10k或10k-40k，更优选为5k；
L 2为
Figure PCTCN2021107555-appb-000073
各r 1独立地为1、2、3、4、5或6，各r 1独立地优选为1、2、3或4，各r 1独立地更优选为3或4；
W为
Figure PCTCN2021107555-appb-000074
Z 2、Z 1、Z 0各自独立地为
Figure PCTCN2021107555-appb-000075
r 1为1、2、3、4、5或6，r 1优选为1、2、3或4，r 1更优选为3或4，各r 2独立地为1、2、3、4、5或6，各r 2独立地优选为1、2、3或4，各r 2独立地更优选为1或2，
Q为N-AC；
N为
Figure PCTCN2021107555-appb-000076
或G，r 0为1、2、3、4、5或6，优选为3、4、5或6，更优选为5或6；
AC为SB7、NPB、SN38、LPT、PCB、DOX、PTX或AXT，优选为PTX或SN38。
在一些实施方案中，L 2为
Figure PCTCN2021107555-appb-000077
在一些实施方案中，Z 2、Z 1、Z 0各自独立地为
Figure PCTCN2021107555-appb-000078
Figure PCTCN2021107555-appb-000079
在一些实施方案中，Z 2为
Figure PCTCN2021107555-appb-000080
r 1为3或4。
在一些实施方案中，Z 2为
Figure PCTCN2021107555-appb-000081
在一些实施方案中，Z 1为
Figure PCTCN2021107555-appb-000082
各r 2独立地为1或2。
在一些实施方案中，Z 1为
Figure PCTCN2021107555-appb-000083
在一些实施方案中，Z 0为
Figure PCTCN2021107555-appb-000084
r 2为1或2。
在一些实施方案中，Z 0为
Figure PCTCN2021107555-appb-000085
在一些实施方案中，N为
Figure PCTCN2021107555-appb-000086
或G。
在一些实施方案中，所述聚乙二醇偶联药物选自：
Figure PCTCN2021107555-appb-000087
在本发明的第七方面，本发明提供了前述的聚乙二醇偶联药物增效剂或其药学上可接受的盐或者前述的组合物在制备药物中的用途，所述药物用于增强治疗和/或预防疾病的疗效。
在一些实施方案中，所述疾病为癌症，所述癌症选自：结肠癌、白血病、淋巴瘤、膀胱癌、骨癌、脑瘤、髓母细胞瘤、胶质瘤、乳腺癌、腺瘤/类癌、肾上腺皮质癌、胰岛细胞癌、子宫颈癌、子宫内膜癌、卵巢癌、结肠直肠癌、皮肤癌、食管癌、眼癌、胆囊癌、胃癌、头颈癌、肝癌、黑色素瘤、卡波氏肉瘤、肾癌、口腔癌、肺癌、鼻咽癌、神经母细胞瘤、卵巢癌、胰腺癌、甲状腺癌、甲状旁腺阴茎癌、前列腺癌、尿道癌、阴道癌、外阴癌、肛门癌、肉瘤，以及所述癌症的转移。
在本发明的第八方面，本发明提供了增强治疗和/或预防疾病的疗效的方法，其包括对有此需要的个体给予有效量的前述的聚乙二醇偶联药物增效剂或其药学上可接受的盐或者前述的组合物。
在一些实施方案中，所述疾病为癌症，所述癌症选自：结肠癌、白血病、淋巴瘤、膀胱癌、骨癌、脑瘤、髓母细胞瘤、胶质瘤、乳腺癌、腺瘤/类癌、肾上腺皮质癌、胰岛细胞癌、子宫颈癌、子宫内膜癌、卵巢癌、结肠直肠癌、皮肤癌、食管癌、眼癌、胆囊癌、胃癌、头颈癌、肝癌、黑色素瘤、卡波氏肉瘤、肾癌、口腔癌、肺癌、鼻咽癌、神经母细胞瘤、卵巢癌、胰腺癌、甲状腺癌、甲状旁腺阴茎癌、前列腺癌、尿道癌、阴道癌、外阴癌、肛门癌、肉瘤，以及所述癌症的转移。
在本发明的第九方面，本发明提供了前述的聚乙二醇偶联药物增效剂或其药学上可接受的盐或者前述的组合物，其用于增强治疗和/或预防疾病的疗效。
在一些实施方案中，所述疾病为癌症，所述癌症选自：结肠癌、白血病、淋巴瘤、膀胱癌、骨癌、脑瘤、髓母细胞瘤、胶质瘤、乳腺癌、腺瘤/类癌、肾上腺皮质癌、胰岛细胞癌、子宫颈癌、子宫内膜癌、卵巢癌、结肠直肠癌、皮肤癌、食管癌、眼癌、胆囊癌、胃癌、头颈癌、肝癌、黑色素瘤、卡波氏肉瘤、肾癌、口腔癌、肺癌、鼻咽癌、神经母细胞瘤、卵巢癌、胰腺癌、甲状腺癌、甲状旁腺阴茎癌、前列腺癌、尿道癌、阴道癌、外阴癌、肛门癌、肉瘤，以及所述癌症的转移。
在本发明的第十方面，本发明提供了前述的组合物(包含抗癌药物)在制备治疗和/或预防疾病的药物中的用途，所述疾病为前述的抗癌药物所治疗的疾病。
在一些实施方案中，所述疾病为癌症，所述癌症选自：结肠癌、白血病、淋巴瘤、膀胱癌、骨癌、脑瘤、髓母细胞瘤、胶质瘤、乳腺癌、腺瘤/类癌、肾上腺皮质癌、胰岛细胞癌、子宫颈癌、子宫内膜癌、卵巢癌、结肠直肠癌、皮肤癌、食管癌、眼癌、胆囊癌、胃癌、头颈癌、肝癌、黑色素瘤、卡波氏肉瘤、肾癌、口腔癌、肺癌、鼻咽癌、神经母细胞瘤、卵巢癌、胰腺癌、甲状腺癌、甲状旁腺阴茎癌、前列腺癌、尿道癌、阴道癌、外阴癌、肛门癌、肉瘤，以及所述癌症的转移。
在本发明的第十一方面，本发明提供了治疗和/或预防疾病的方法，其包括对有此需要的个体给予有效量的前述的组合物(包含抗癌药物)，所述疾病为前述的抗癌药物所治疗的疾病。可调整给药方案以提供最佳所需响应。例如，可给药单次推注，可随时间给药数个分剂量，或可如治疗情况的急需所表明而按比例减少或增加剂量。要注意，剂量值可随要减轻的病况的类型及严重性而变化，且可包括单次或多次剂量。要进一步理解，对于任何特定个体，具体的给药方案应根据个体需要及给药组合物或监督组合物的给药的人员的专业判断来随时间调整。
在一些实施方案中，所述疾病为癌症，所述癌症选自：结肠癌、白血病、淋巴瘤、膀胱癌、骨癌、脑瘤、髓母细胞瘤、胶质瘤、乳腺癌、腺瘤/类癌、肾上腺皮质癌、胰岛细胞癌、子宫颈癌、子宫内膜癌、卵巢癌、结肠直肠癌、皮肤癌、食管癌、眼癌、胆囊癌、胃癌、头颈癌、肝癌、黑色素瘤、卡波氏肉瘤、肾癌、口腔癌、肺癌、鼻咽癌、神经母细胞瘤、卵巢癌、胰腺癌、甲状腺癌、甲状旁腺阴茎癌、前列腺癌、尿道癌、阴道癌、外阴癌、肛门癌、肉瘤，以及所述癌症的转移。
在本发明的第十二方面，本发明提供了前述的组合物(包含抗癌药物)，其用于治疗和/或预防疾病，所述疾病为前述的抗癌药物所治疗的疾病。
在一些实施方案中，所述疾病为癌症，所述癌症选自：结肠癌、白血病、淋巴瘤、膀胱癌、骨癌、脑瘤、髓母细胞瘤、胶质瘤、乳腺癌、腺瘤/类癌、肾上腺皮质癌、胰岛细胞癌、子宫颈癌、子宫内膜癌、卵巢癌、结肠直肠癌、皮肤癌、食管癌、眼癌、胆囊癌、胃癌、头颈癌、肝癌、黑色素瘤、卡波氏肉瘤、肾癌、口腔癌、肺癌、鼻咽癌、神经母细胞瘤、卵巢癌、胰腺癌、甲状腺癌、甲状旁腺阴茎癌、前列腺癌、尿道癌、阴道癌、外阴癌、肛门癌、肉瘤，以及所述癌症的转移。
本发明中，癌症是指细胞增殖性疾病状态，包括但不限于：结肠癌、白血 病、淋巴瘤、膀胱癌、骨癌、脑瘤、髓母细胞瘤、胶质瘤、乳腺癌、腺瘤/类癌、肾上腺皮质癌、胰岛细胞癌、子宫颈癌、子宫内膜癌、卵巢癌、结肠直肠癌、皮肤癌、食管癌、眼癌、胆囊癌、胃癌、头颈癌、肝癌、黑色素瘤、卡波氏肉瘤、肾癌、口腔癌、肺癌、鼻咽癌、神经母细胞瘤、卵巢癌、胰腺癌、甲状腺癌、甲状旁腺阴茎癌、前列腺癌、尿道癌、阴道癌、外阴癌、肛门癌、肉瘤等，包括前述癌症的转移。
本发明中，“个体”包括人或非人动物。示例性人个体包括患有疾病(例如本文所述的疾病)的人个体(称为患者)或正常个体。本发明中，“非人动物”包括所有脊椎动物，例如非哺乳动物(例如鸟类、两栖动物、爬行动物)和哺乳动物，例如非人灵长类、家畜和/或驯化动物(例如绵羊、犬、猫、奶牛、猪等)。
如本文中所使用的，术语“有效量”指被给药后会在一定程度上缓解所治疗病症的一或多种症状的化合物的量。
如本文中所使用，术语“治疗”意指逆转、减轻、抑制这样的术语所应用的病症或病况或者这样的病症或病况的一或多种症状的进展，或预防这样的病症或病况或者这样的病症或病况的一或多种症状。
本发明的聚乙二醇偶联药物增效剂，以氨基酸或多肽等为连接链，以带有氨基的二羧酸或多羧酸(例如带有两个羧基的天然氨基酸)或者带有二氨基或多氨基的羧酸(例如带有两个氨基的天然氨基酸)或者多羧酸为连接桥，通过形成酰胺键将多个PPT-iRGD或MI-AH-PPT-iRGD偶联在一起。在某些实施方案中，活化的PEG通过羧基与主链上的氨基发生反应，形成酰胺键。在某些实施方案中，PEG通过末端的氨基与主链上的羧基发生反应，形成酰胺键。在某些实施方案中，PEG的分子量包含其末端的氨基(即带有活性基团的PEG衍生物)。
本发明的聚乙二醇偶联药物，以氨基酸或多肽等为连接链，以带有氨基的二羧酸或多羧酸(例如带有两个羧基的天然氨基酸)或者带有二氨基或多氨基的羧酸(例如带有两个氨基的天然氨基酸)或者多羧酸为连接桥，通过形成酰胺键将多个相同或不同的药物分子偶联在一起。药物的种类、比例和载药量可以调节。在某些实施方案中，活化的PEG通过羧基与主链上的氨基发生反应，形成酰胺键。在某些实施方案中，PEG的分子量包含其末端的氨基(即带有活性基团的PEG衍生物)。
本发明的聚乙二醇偶联药物中，适于与聚乙二醇偶联的活性成分可以是带有至少一个氨基、羟基、羧基或者酰基的药物分子，例如带有至少一个氨基、羟基、羧基或者酰基的具有抗肿瘤活性的药物分子，例如SB7、NPB、SN38、LPT、PCB、DOX、PTX或AXT，其代表的含义如下：
缩写 名称 CAS号或结构式
LPT 拉帕替尼 231277-92-2
PCB 帕布惜利布 571190-30-2
SB7 SB-743921 940929-33-9
NPB Niraparib(MK-4827) 1038915-60-4
SN38 7-乙基-10-羟基喜树碱 86639-52-3
DOX 阿霉素 23214-92-8
PTX 紫杉醇 33069-62-4
AXT Axitinib 319460-85-0
PPT-iRGD的结构式为
Figure PCTCN2021107555-appb-000088
PPT-iRGD与聚乙二醇偶联药物增效剂整体结构的其他部分的连接位点为末端巯基
Figure PCTCN2021107555-appb-000089
即
Figure PCTCN2021107555-appb-000090
所示位置。
MI-AH-PPT-iRGD的结构式为
Figure PCTCN2021107555-appb-000091
MI-AH-PPT-iRGD与聚乙二醇偶联药物增效剂整体结构的其他部分的连接位点为马来酰亚胺基团
Figure PCTCN2021107555-appb-000092
即
Figure PCTCN2021107555-appb-000093
所示位置。在一些实施方案中，MI-AH-PPT-iRGD在 马来酰亚胺基团上与另外的巯基进行加成连接。
另外，以下药物分子与聚乙二醇偶联药物整体结构的其他部分的连接位点如下表所示，具体即
Figure PCTCN2021107555-appb-000094
所示位置。
Figure PCTCN2021107555-appb-000095
如本文中所使用，“PEG”为聚乙二醇的缩写，是指重复单元为-CH 2CH 2O-的均聚物，包括单臂聚乙二醇、多臂聚乙二醇及其衍生物，例如端基带有氨基或羧基等反应官能团的衍生物。本发明的结构式中，聚乙二醇重复单元下角标的字母n代表聚乙二醇的聚合度。
如本文中所使用，除非以其他方式明确指出，在本文中通篇采用的描述方式“各…独立地为/选自”和“…和…各自独立地为/选自”可以互换，均应做广义理解，其既可以是指在不同基团中，相同符号之间所表达的具体选项之间互相不影响，也可以表示在相同的基团中，相同符号之间所表达的具体选项之间互相不影响。
如本文中所使用，本发明的化合物的“药学上可接受的盐”包括化合物的酸加成盐及碱加成盐。例如盐酸盐、六氟磷酸盐、葡甲胺盐等。
如本文中所使用，结构式中的波浪线
Figure PCTCN2021107555-appb-000096
意指其他基团与结构式所代表的结构发生键连的位置。
附图说明
图1表示本发明实施例的各组肿瘤增长趋势示意图。
图2表示本发明实施例的各组肿瘤重量抑制率示意图。
具体实施方式
下面将结合实施例对本发明的实施方案进行详细描述，但是本领域技术人员将会理解，下列实施例仅用于说明本发明，而不应视为限定本发明的范围。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。
实施例中各缩写的含义如下：
Figure PCTCN2021107555-appb-000097
部分原料的来源和结构如下：
M-NH 2-10K.HCl
键凯，mPEG-CH 2CH 2-NH 2HCl
M-NH 2-5K.HCl
键凯，mPEG-CH 2CH 2-NH 2HCl
M-SCM-5K
键凯，
Figure PCTCN2021107555-appb-000098
M-SCM-10K
键凯，
Figure PCTCN2021107555-appb-000099
下面结合具体的实施例对本发明进行进一步的解释说明。
实施例1 化合物的制备
45-164的合成：
Figure PCTCN2021107555-appb-000100
Figure PCTCN2021107555-appb-000101
45-114
Figure PCTCN2021107555-appb-000102
称取Boc-L-Lys(Fmoc)-OH(20g,42.6857mmol，购于阿拉丁)、Gly-OBn(14.4g,42.6857mmol，购于innochem)、HBTU(24g,64.0286mmol)、HOBT(8.65g,64.0286mmol)投入500mL反应瓶中，加入DMF(40mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(31.7mL，192.085mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，蒸干得产品26克。
45-115
Figure PCTCN2021107555-appb-000103
称取化合物45-114(26g，42.6857mmol)，加入二氯甲烷(20mL)，再加入TFA(14mL，190.2855mmol)，超声溶解，搅拌反应。反应结束，反应液浓缩，再用乙酸乙酯(100mL)溶解，加饱和氯化钠溶液，分离有机相，水相用乙酸乙酯萃取3次(50mL×3)，直至水相无产品，合并有机相，再用饱和食盐水清洗一次(50mL)，浓缩蒸干得产品22g。
45-124
Figure PCTCN2021107555-appb-000104
称取Boc-L-Lys(Boc)-OH(7.3g,21.334mmol，购于Aladdin)、45-115(11g,21.334mmol)、HBTU(12.1363g,32.0016mmol)、HOBT(4.3241g,32.0016mmol)投入500mL反应瓶中，加入DMF(80mL)溶解，-5℃条件下搅拌30分钟后。 缓慢滴加DIEA(15mL，96.003mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，干法上样，柱层析，用50％-100％乙酸乙酯/石油醚梯度洗脱得产品15克，产率83％。
45-126
Figure PCTCN2021107555-appb-000105
称取化合物45-124(15g，17.7725mmol)，加入二氯甲烷(30mL)，再加入TFA(39mL，533.1754mmol)，超声溶解，搅拌反应。反应结束，反应液浓缩，再用乙酸乙酯(100mL)溶解，加饱和碳酸氢钠溶液至水相显碱性，分离有机相，水相用乙酸乙酯萃取3次(150mL×3)，直至水相无产品，合并有机相，再用饱和食盐水清洗一次(150mL)，浓缩蒸干得产品11.44g。
45-127
Figure PCTCN2021107555-appb-000106
称取Boc-L-Lys(Boc)-OH(13.022g,37.5911mmol)、45-126(11.4g,17.0868mmol)、HBTU(19.5538g,51.5604mmol)、HOBT(6.9668g,51.5604mmol)投入500mL反应瓶中，加入DMF(80mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(25.4mL，153.7812mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，干法上样，柱层析，用1％-55甲醇/二氯甲烷梯度洗脱得产品18克，产率81％。
45-135
Figure PCTCN2021107555-appb-000107
取反应物45-127(1.1g,0.8458mmol)置于250mL烧瓶中，加DMF溶液(50mL)，加入吗啉(1.4745mL,16.9155mmoL)，常温搅拌反应，3小时后，反应结束。饱和食盐水(150mL)和乙酸乙酯(200mL)，分离有机相，水相用乙酸乙酯萃取3次(150mL×3)，水相中无产品，合并有机相，用饱和食盐水清洗2次(150mL×2)，有机相蒸干得产品0.912克。
45-57
Figure PCTCN2021107555-appb-000108
称季戊四醇(10g,73.4484mmol)投500mL两颈反应瓶中，加四氢呋喃(20mL)超声均匀，置于0℃条件下搅拌，通入氮气保护后，加叔丁醇钾(352mL,352.55mmol)，℃下搅拌2小时后，加入溴乙酸苄酯(55mL,352.55mmol)，再搅拌3小时后移至常温下反应。反应结束，用纯水与乙酸乙酯萃取处理反应液，浓缩有机相，干法上样，柱层析。用1％-2％乙酸乙酯/石油醚梯度洗脱得产品15g，产率28％。
45-64
Figure PCTCN2021107555-appb-000109
称45-57(0.76g,1.0538mmol)与10％Pd/C(100mg)投入氢化反应釜中，缓慢加入DMF(30mL)搅拌溶解，通入氢气(300psi)，常温搅拌过夜。次日，用装有硅藻土的砂芯漏斗抽滤反应液，除去Pd/C，得45-64的DMF溶液，直接投下一步反应。
45-136
Figure PCTCN2021107555-appb-000110
称取45-135(0.9121g,0.8458mmol)、HBTU(0.4009g,1.0572mmol)、HOBT(0.1388g,10572mmol)投入250mL反应瓶中，加入45-64(0.1762mmol)的DMF溶液溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(0.5242mL，3.1716mmol)，-5℃条件下反应过夜。反应结束，在反应液中加纯水，抽滤，干法上样，柱层析。用1％-5％甲醇/二氯甲烷洗脱得产品0.4g，产率50％。
1H-NMR(600MHz,DMSO-d 6)δ8.37-8.33(m,2H),7.98–7.81(m,13H),7.72-7.70(m,9H),7.63-7.62(m,1H),7.40-7.30(m,24H),6.89-6.86(m,2H),6.74-6.69(m,6H),5.11-5.10(m,4H),4.55-4.54(m,3H),4.24-4.16(m,6H),3.99–3.78(m,12H),3.65-3.62(m,6H),3.37-3.33(s,48H),3.08–2.82(m,42H),2.71-2.68(m,25H),2.57-2.53(m,10H),1.68–1.14(m,161H).
45-138
Figure PCTCN2021107555-appb-000111
称45-136(0.35g,0.0759mmol)与10％Pd/C(75mg)投入氢化反应釜中，缓慢加入DMF(50mL)搅拌溶解，通入氢气(20psi)，常温搅拌过夜。次日，用装有硅藻土的砂芯漏斗抽滤反应液，除去Pd/C，得产品的DMF溶液，直接投下一步反应。
45-139
Figure PCTCN2021107555-appb-000112
称取M-NH 2HCl-10K(3.3679g,0.3188mmol，购于键凯)、HBTU(0.1727g,0.4554mmol)、HOBT(0.0615g,0.4554mmol)投入250mL反应瓶中，加入45-138(0.0667mmol)的DMF溶液溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(0.1766mL，1.3662mmol)，-5℃条件下反应1小时后移至常温下低速避光反应。反应结束，在反应液中加甲基叔丁基醚，抽滤，干法上样，柱层析。用5％-8％甲醇/二氯甲烷洗脱得产品1.2g。
1H-NMR(600MHz,DMSO-d 6)δ9.04-9.02(m,2H),7.96-7.92(m,8H),7.89-7.77(m,10H),7.37-7.34(m,12H),6.78-6.61(m,9H),4.53-4.52(m,4H),3.87-3.85(m,9H),3.64-3.60(m,46H),3.51-3.49(m,3683H),3.41-3.39(m,18H),3.24-3.23(m,6H),2.90-2.88(m,12H),2.74-2.72(m,14H),2.62-2.60(m,29H),2.39-2.37(m,8H),1.38–1.34(m,85H),1.27–1.20(m,84H).
45-154
Figure PCTCN2021107555-appb-000113
称取化合物45-139(0.4g，0.0146mmol)，加入二氯甲烷(10mL)，再加入TFA(0.25mL，3.4983mol)，超声溶解，搅拌反应。反应结束，在反应液中加甲醇(30mL)，加过量碳酸钾，搅拌30分钟，抽滤，滤液中加硅胶粉蒸干，上样，柱层析。用6％-50％甲醇/二氯甲烷梯度洗脱得产品0.2g，产率54％。
45-156
Figure PCTCN2021107555-appb-000114
称取45-154(0.2g,0.0044mmol)、6-马来酰亚胺基己酸(0.0189g,0.0895mmol)、HBTU(0.0400g,0.1056mmol)、HOBT(0.0143g,0.1056mmol)投入250mL反应瓶中，DMF溶液溶解，-5℃条件下避光搅拌30分钟后。缓慢滴加DIEA(0.0523mL，0.3168mmol)，-5℃条件下反应1小时后移至常温下避光反应。反应结束，在反应液中加甲基叔丁基醚倾倒多次得产品0.1g，产率50％。
45-164
Figure PCTCN2021107555-appb-000115
称取45-156(0.4477g,0.0094mmol)、PPT-iRGD(0.3g,0.1874mmol，购于丹港生物科技)、投入250mL反应瓶中，加DMSO(50mL)溶液溶解，40℃条件下避光搅拌2天，常温搅拌3天。反应结束，在反应液中加甲基叔丁基醚，倾倒，固体不用10％甲醇/二氯甲烷溶解，加硅胶粉(5g)，干法上样，柱层析。用5％-10％甲醇/二氯甲烷洗脱得产品0.3g，产率44％。
1H-NMR(600MHz,DMSO-d 6)δ9.13-9.12(m,6H),8.40-7.83(m,255H),7.40-7.26(m,94H),7.13-7.12(m,6H),7.01–6.88(m,3H),6.70-6.68(m,4H),4.54-4.52(m,52H),4.26-4.24(m,83H),3.78-3.76(m,131H),3.51-3.49(m,4200H),3.09-3.07(m,178H),2.89-2.86(m,225H),2.71-2.69(m,212H),2.37-2.35(m,22H),2.16-1.97(m,52H),1.81-1.76(m,102H),1.55-1.52(m,174H),1.33-1.28(m,288H),0.85-0.81(m,192H).
化合物49-166的合成(抗癌药)
Figure PCTCN2021107555-appb-000116
Figure PCTCN2021107555-appb-000117
Figure PCTCN2021107555-appb-000118
Figure PCTCN2021107555-appb-000119
39-80
Figure PCTCN2021107555-appb-000120
称取Boc-Glu-OH(5.0g，20.22mmol，购于阿拉丁)、H-Glu(OBzl)-OBzl·TsOH(21.2g，42.46mmol，购于Ark Pharm)、HOBT(8g,60.66mmol)、HBTU(23g,60.66mmol)投入250mL反应瓶中，加DMF溶液(80mL)溶解，超声使反应物完全溶解，-5℃条件下搅拌30分钟，缓慢滴加DIEA(30mL，181mmol)，低温反应至结束。反应结束后，取出反应液加入去离子水(100mL)，用乙酸乙酯萃取多次(100mL×3)，合并有机相，有机相加入饱和氯化钠溶液(100mL)清洗两次，最后浓缩蒸干。干法上样，柱层析，用40％乙酸乙酯/石油醚-50％乙酸乙酯/石油醚梯度洗脱收集产品，浓缩蒸干。
39-84
Figure PCTCN2021107555-appb-000121
取39-80(19.2mmol)，加入二氯甲烷(5mL)，再加入TFA(14mL，192mmol)，超声使完全溶解，用磨口玻璃塞，常温搅拌反应。反应结束后，取出反应液加入饱和碳酸氢钠溶液(300mL)，用乙酸乙酯萃取多次(100mL×3)，合并有机相，有机相加入饱和氯化钠溶液(100mL)清洗两次，最后浓缩蒸干。
47-96
Figure PCTCN2021107555-appb-000122
将7-乙基-10-羟基喜树碱(15.00g，38.23mmol,简写为SN38)置于1000ml圆底烧瓶中，加入二氯甲烷150ml使之溶解，加入叔丁基二苯基氯硅烷(59.64ml，229.36mmol，购于韶远)、三乙胺(31.88ml，229.36mmol)，反应在37℃油浴下搅拌过夜。反应结束后,将反应液蒸至粘稠状，加入正己烷(150ml)将产物沉降至固体，抽滤、烘干，得到产品(23.15g，96％)。
47-97
Figure PCTCN2021107555-appb-000123
将47-96(23.15g,36.70mmol)、Boc-Gly-OH(8.71g，49.70mmol，购于阿拉丁)、DMAP(0.94g,7.65mmol)置于1000mL圆底烧瓶中，然后加入二氯甲烷(150mL)，使其溶解，将混合液置于0℃条件下搅拌30分钟。然后分三次共加入DCC(39.41g,191.15mmol)，每次加入DCC间隔时间为30min，结束后0℃下反应2小时，然后将反应装置放到室温条件下搅拌过夜反应。反应结束后，加入正己烷(200mL)和石油醚(50mL)沉降，重复3次，过滤得到固体产物，放入真空箱中干燥，得到产品(27.53g，94％)。
47-98
Figure PCTCN2021107555-appb-000124
将47-97(27.53g，34.50mmol)置于1000ml圆底烧瓶中，加入二氯甲烷(50ml)溶解后，加入三氟乙酸(28.40ml，382.30mmol)，将反应至于室温下进行。反应结束后,取出反应液加入去离子水(200mL)，用乙酸乙酯(100mL×3)萃取，合并有机相，有机相加入饱和氯化钠溶液(200mL×2)清洗，浓缩，加入硅胶粉，蒸干至粉末状，柱层析，用(1％-3％CH 3OH,其余CH 2Cl 2)梯度洗脱，收集浓缩，得到产品(16.98g，72％)。
29-242
Figure PCTCN2021107555-appb-000125
将Fmoc-Lys(Boc)-OH(5.0g，10.6714mmol，购于阿拉丁)、H-Gly-OBn(3.7802g，11.2050mmol,购于innochem)、HBTU(6.0705g,16.0072mmol)、HOBT(2.1630g，16.0072mmol)加入到500ml烧瓶中，加入DMF(50mL)使其溶解，置于0摄氏度下搅拌反应30分钟。再缓慢滴加DIEA(7.9371mL，48.0215mmol)，继续在0摄氏度下搅拌反应过夜。反应结束后,取出反应液加入去离子水(200mL)，用乙酸乙酯(100mL×3)萃取，合并有机相，有机相加入饱和氯化钠溶液(200mL)清洗两次，浓缩蒸干，真空箱干燥，得到粗产品。29-243
Figure PCTCN2021107555-appb-000126
将29-242(6.57g，10.6714mmol)加入500ml烧瓶中，加入适量的二氯甲烷和TFA(7.9248mL，106.714mmol)使之溶解，反应在室温下搅拌过夜。反应结束后,取出反应液蒸至油状，加饱和碳酸氢钠溶液调节PH至碱性，再用乙酸乙酯(100mL×3)萃取，合并有机相，有机相加入饱和氯化钠溶液(200mL)清洗两次，浓缩蒸干，真空箱干燥，得到粗产品。
29-245
Figure PCTCN2021107555-appb-000127
将Boc-Lys(Boc)-OH(4.2805g，11.7385mmol，购于Ark Pharm)、29-243(5.50g，10.6714mmol)、HBTU(6.0705g,16.0072mmol)、HOBT(2.1630g、16.0072mmol)加入到500ml烧瓶中，加入DMF(50mL)使其溶解，置于0摄氏度下搅拌反应30分钟。再缓慢滴加DIEA(7.9371mL，48.0215mmol)，继续在0摄氏度下反应过夜。反应结束后,取出反应液加入去离子水(200mL)，用乙酸乙酯(100mL×3)萃取，合并有机相，有机相加入饱和氯化钠溶液(200mL)清洗两次，浓缩蒸干，真空箱干燥，得到粗产品。
29-246
Figure PCTCN2021107555-appb-000128
将吗啉(9.24mL，106.714mmol)和DMF(10mL)加入到装有29-245(9.0g，10.6714mmol)的500ml烧瓶中，使之溶解，反应在室温下搅拌1小时。反应结束后,取出反应液加入去离子水(200mL)，用乙酸乙酯(100mL×3)萃取，合并有机相，有机相加入饱和氯化钠溶液(200mL)清洗两次，浓缩，加入硅胶粉，蒸干至粉末状，柱层析，用(1％氨水，4％-5％甲醇,其余二氯甲烷)梯度洗脱，收集浓缩，得到产品29-246：3.7g，产率56％。
29-248
Figure PCTCN2021107555-appb-000129
将38-120(0.39g，1.0966mmol)、29-246(3.0g，4.8249mmol)、HBTU(2.4951g,6.5795mmol)、HOBT(0.8891g,6.5795mmol)加入到500ml烧瓶中，加入DMF(50mL)使其溶解，置于-5摄氏度下搅拌反应30分钟。再缓慢滴加DIEA(3.2624mL，19.7384mmol)，在-5摄氏度下搅拌反应1小时后，移至室温下搅拌过夜。反应结束后,取出反应液加入去离子水(200mL)，用乙酸乙酯(100mL×3)萃取，合并有机相，有机相加入饱和氯化钠溶液(200mL)清洗两次，浓缩，加入硅胶粉，蒸干至粉末状，柱层析，用(50％-80％乙酸乙酯,其余石油醚)梯度洗脱，收集浓缩，得到产品(1.6g，53％)。
49-79
Figure PCTCN2021107555-appb-000130
将29-248(0.52g，0.1878mmol)和10％Pd/C(0.0,5g)投入到氢化反应釜中，然后加入DMF(30mL)使其溶解，封闭氢化反应装置，进行三抽三充后使氢化反应装置上的压力读数为0.18MPa，然后在常温下过夜反应。反应结束,反应液用硅藻土过滤，滤饼用DMF(20mLⅹ3)清洗，得到产品的DMF溶液，作为下一部反应原料。
49-80
Figure PCTCN2021107555-appb-000131
将49-79(0.45g，0.1878mmol)、M-NH 2HCl-5K(4.91g，0.9389mmol，购于键凯)、HBTU(0.43g,1.1237mmol)、HOBT(0.15g,1.1267mmol)加入到500ml烧瓶中，加入DMF(60mL)使其溶解，置于-5摄氏度下搅拌反应30分钟。再缓慢滴加DIEA(0.56mL，3.3780mmol)，在-5摄氏度下搅拌反应1小时后，移至室温下低速避光搅拌三天。反应结束，用正己烷(100mL)震荡，倒掉上清液，重复上述操作三次，再加甲基叔丁基醚(80mL)和少量正己烷(10mL)震荡，倒掉上清液，重复三次，反应液中粉末状固体析出，抽滤，滤饼用甲基叔丁基醚(40mL x 3)清洗，收集滤饼，然后用甲醇/二氯甲烷(1:4)溶液(100mL)溶解，加入硅胶粉(20g)，蒸干至粉末状，干法上样，柱层析，用1％氨水和3％-7％甲醇的二氯甲烷混合溶液梯度洗脱，收集浓缩，真空烘箱干燥，得到产品(3.2g，73.73％)。
49-153
Figure PCTCN2021107555-appb-000132
将49-80(3.2g，0.1383mmol)加入250mL烧瓶中，加入二氯甲烷(5mL)和TFA(0.82mL,11.0640mmol)使之溶解，反应在室温下低速避光搅拌过夜。反应结束，用旋转蒸发仪将反应液蒸至油状，再加入甲基叔丁基醚(60mL)，反应液中粉末状固体析出，抽滤，滤饼用甲基叔丁基醚(40mL x 3)清洗，收集滤饼，然后用甲醇/二氯甲烷(1:4)溶液(100mL)溶解，加入硅胶粉(20g)，蒸干至粉末状，干法上样，柱层析，用3％-12％甲醇的二氯甲烷混合溶液梯度洗脱，收集浓缩，真空烘箱干燥，得到产品(1.74g，56.35％)。
49-157
Figure PCTCN2021107555-appb-000133
将39-84(5.16g，6.7433mmol)、单叔丁基琥珀酸(1.40g，8.0920mmol，购于Accela)、HBTU(3.84g,10.1149mmol)、HOBT(1.36g，10.1149mmol)加入500ml烧瓶中，加入DMF(50mL)使其溶解，置于-5摄氏度下搅拌反应30分钟。再缓慢滴加DIEA(10.03mL，60.6897mmol)，在-5摄氏度下搅拌反应1小时后，移至室温下搅拌过夜。反应结束后,取出反应液加入去离子水(200mL)，用乙酸乙酯(100mL×3)萃取，合并有机相，有机相加入饱和氯化钠溶液(200mL)清洗两次，浓缩，加入硅胶粉，蒸干至粉末状，柱层析，用(50％-90％乙酸乙酯,其余石油醚)梯度洗脱，收集浓缩，得到产品(5.66g，90.99％)。
49-158
Figure PCTCN2021107555-appb-000134
将49-157(2.8g，3.0367mmol)和10％Pd/c(0.08g)投入到氢化反应釜中，然后加入DMF(30mL)使其溶解，封闭氢化反应装置，进行三抽三充后使氢化反应装置上的压力读数为0.18MPa，然后在常温下过夜反应。反应结束,反应液用硅藻土过滤，滤饼用DMF(20mLⅹ3)清洗，得到产品的DMF溶液,作为下一部反应原料。
49-159
Figure PCTCN2021107555-appb-000135
将49-158(1.71g，3.0367mmol)、47-98(8.77g，12.7541mmol)、HBTU (6.90g,18.2202mmol)、HOBT(2.46g,18.2202mmol)加入到500ml烧瓶中，加入DMF(60mL)使其溶解，置于-5摄氏度下搅拌反应30分钟。再缓慢滴加DIEA(13mL，78.9542mmol)，在-5摄氏度下搅拌反应1小时后，移至室温下搅拌过夜。反应结束，用正己烷(100mL)震荡，倒掉上清液，重复上述操作三次，再加甲基叔丁基醚(80mL)和少量正己烷(10mL)震荡，倒掉上清液，重复三次，反应液中粉末状固体析出，抽滤，滤饼用甲基叔丁基醚(40mL x 3)清洗，收集滤饼，然后用甲醇/二氯甲烷(1:4)溶液(100mL)溶解，加入硅胶粉(60g)，蒸干至粉末状，干法上样，柱层析，用3％-7％甲醇的二氯甲烷混合溶液梯度洗脱，收集浓缩，真空烘箱干燥，得到产品(11.3g，超产)。
49-161
Figure PCTCN2021107555-appb-000136
将49-159(9.84g，3.0367mmol)加入250mL烧瓶中，加入二氯甲烷(8mL)和TFA(8mL)使之溶解，反应在室温下搅拌过夜。反应结束，用旋转蒸发仪将反应液蒸至油状，再加入甲基叔丁基醚(60mL)，反应液中粉末状固体析出，抽滤，滤饼用甲基叔丁基醚(40mL x 3)清洗，收集滤饼，然后用甲醇/二氯甲烷(1:4)溶液(100mL)溶解，加入硅胶粉(60g)，蒸干至粉末状，干法上样，柱层析，用1％-4％甲醇的二氯甲烷混合溶液梯度洗脱，收集浓缩，真空烘箱干燥，得到产品(2.7g，27.92％)。
49-162
Figure PCTCN2021107555-appb-000137
将49-153(1.19g，0.0535mmol)、49-161(1.5g，0.4710mmol)、HBTU(0.24g,0.6422mmol)、HOBT(0.08g,0.6422mmol)加入到500ml烧瓶中，加入DMF(60mL)使其溶解，置于-5摄氏度下搅拌反应30分钟。再缓慢滴加DIEA(0.60mL，3.6395mmol)，在-5摄氏度下搅拌反应1小时后，移至室温下低速避光搅拌过夜。反应结束，加入正己烷(100mL)震荡，倒掉上清液，重复上述操作三次，再加甲基叔丁基醚(80mL)和少量正己烷(10mL)震荡，倒掉上清液，重复三次，反应液中粉末状固体析出，抽滤，滤饼用甲基叔丁基醚(40mL x 3)清洗，收集滤饼，然后用甲醇/二氯甲烷(1:4)溶液(100mL)溶解，加入硅胶粉(20g)，蒸干至粉末状，干法上样，柱层析，用3％-15％甲醇的二氯甲烷混合溶液梯度洗脱，收集浓缩，真空烘箱干燥，得到产品(0.95g，37.25％)。
49-166
Figure PCTCN2021107555-appb-000138
向装有49-162(0.95g，0.0199mmol)的500ml烧瓶中，加入THF(10ml)和稀盐酸(10ml，0.05mmol/L)，超声使之溶解，再加入TBAF(0.5g,1.9133mmol)，反应在室温下避光搅拌3小时。反应结束后，将反应液蒸干，用DMF(5ml)溶解，再用异丙醇沉降，重复三次；再用无水乙醇和少量二氯甲烷溶解，加甲基叔丁基醚沉降，重复三次，收集固体，真空箱干燥，得到产品(0.75g，93.75％)。
1H-NMR(600MHz,DMSO-d 6)δ8.44–8.22(m,24H),8.05–7.89(m,46H),7.44–7.27(m,78H),5.53–5.34(m,50H),5.31–5.08(m,52H),4.55–4.44(m,15H),4.29–4.13(m,31H),4.11–3.89(m,71H),3.89–3.80(m,23H),3.78–3.60(m,82H),3.55–3.44(m,1892H),3.11–2.75(m,100H),2.36–2.21(m,28H),2.20–1.97(m,91H),1.61–1.49(m,34H),1.31–1.16(m,136H),0.97–0.91(m,30H),0.90–0.76(m,79H)。
38-161的合成
Figure PCTCN2021107555-appb-000139
Figure PCTCN2021107555-appb-000140
Figure PCTCN2021107555-appb-000141
38-137
Figure PCTCN2021107555-appb-000142
称取Fmoc-Glu-OH(2.974g,8.05mmol，购于阿拉丁)、NH 2-Glu-OtBu(5g,16.90mmol，购于innochem)、HBTU(9.16g,24.15mmol)、HOBT(3.26g,24.15mmol)投入500mL反应瓶中，加入DMF(10mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(11.97mL，72.45mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，加无水硫酸钠干燥，抽滤，浓缩有机相，蒸干得产品7克。
38-139
Figure PCTCN2021107555-appb-000143
取反应物38-137(6.9g,8.05mmol)置于250mL烧瓶中，加DMF溶液(50mL)，加入吗啉(14mL,161mmoL)，常温搅拌反应，3小时后，反应结束。饱和食盐水(150mL)和乙酸乙酯(200mL)，分离有机相，水相用乙酸乙酯萃取3次(150mL×3)，水相中无产品，合并有机相，用饱和食盐水清洗2次(150mL×2)，有机相蒸干得产品5克。
38-142
Figure PCTCN2021107555-appb-000144
称取38-139(5g,8.05mmol),投入250mL反应瓶中，加DMF(50mL)溶解，在-5℃搅拌条件下加入DIEA(6.6526mL,40.25mmol)，反应30分钟后快速加入丁二酸酐(2.4137g,24.15,mmol)，低温反应结束，反应结束，用去离子水与乙酸乙酯萃取处理，有机相用饱和食盐水清洗2次，浓缩有机相，蒸干得产品5g,产率86％。
38-143
Figure PCTCN2021107555-appb-000145
称取38-142(5g,6.8506mmol)、45-115(3.53g,6.8506mmol)、HBTU(3.897g,10.2759mmol)、HOBT(1.3885g,10.2759mmol)投入500mL反应瓶中，加入DMF(10mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(5.0952mL，30.8277mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，蒸干得产品7克，产率83.3％。
38-144
Figure PCTCN2021107555-appb-000146
取反应物38-143(7g,5.7029mmol)置于250mL烧瓶中，加DMF溶液(50mL)，加入吗啉(9..9368mL,114.0585mmol)，常温搅拌反应，3小时后，反应结束。饱和食盐水(150mL)和乙酸乙酯(200mL)，分离有机相，水相用乙酸乙酯萃取3次(150mL×3)，水相中无产品，合并有机相，用饱和食盐水清洗2次(150mL×2)，有机相蒸干，柱层析，用1％-4％甲醇/二氯甲烷梯度洗脱得产品4克，产率70％。
21-221
Figure PCTCN2021107555-appb-000147
称赤藓醇(4.5g,36.849mmol)投500mL两颈反应瓶中，加四氢呋喃(60mL)超声均匀，置于0℃条件下搅拌，通入氮气保护后，加叔丁醇钾(200mL,184.245mmol)，0℃下搅拌2小时后，加入溴乙酸苄酯(29.187mL,184.245mmol)，再搅拌3小时后移至常温下反应。反应结束，用纯水与乙酸乙酯萃取处理反应液，浓缩有机相，干法上样，柱层析。用1％-2％乙酸乙酯/石油醚梯度洗脱得产品5g，产率20％。
38-145
Figure PCTCN2021107555-appb-000148
称21-221(0.64g,0.9793mmol)与10％Pd/C(100mg)投入氢化反应釜中，缓慢加入DMF(30mL)搅拌溶解，通入氢气(300psi)，常温搅拌过夜。次日，用装有硅藻土的砂芯漏斗抽滤反应液，除去Pd/C，得产品的DMF溶液，直接投下一步反应。
38-146
Figure PCTCN2021107555-appb-000149
称取38-144(4g,3.9793mmol)、HBTU(9.0546g,23.8758mmol)、HOBT(3.2261g,23.8758mmol)投入250mL反应瓶中，加入38-145(0.9043mmol)的DMF溶液溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(11.8354mL，71.6274mmol)，-5℃条件下反应过夜。反应结束，在反应液中加纯水，抽滤，干法上样，柱层析。用1％-5％甲醇/二氯甲烷洗脱得产品2.5g，产率74.34％。38-148
Figure PCTCN2021107555-appb-000150
称38-146(0.29g,0.0676mmol)与10％Pd/C(50mg)投入氢化反应釜中，缓慢加入DMF(30mL)搅拌溶解，通入氢气(300psi)，常温搅拌过夜。次日，用装有硅藻土的砂芯漏斗抽滤反应液，除去Pd/C，得产品的DMF溶液，直接投下一步反应。
38-149
Figure PCTCN2021107555-appb-000151
称取N-Boc-乙二胺(0.0656g,0.4092mmol)、HBTU(0.2116g,0.5580mmol)、HOBT(0.0754g,0.5580mmol)投入250mL反应瓶中，加入38-148(0.0930mmol)的DMF溶液溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(0.27mL，1.67mmol)，-5℃条件下反应过夜。反应结束，在反应液中加纯水，抽滤，干法上样，柱层析。用1％-5％甲醇/二氯甲烷洗脱得产品0.2g，产率50％。
38-152
Figure PCTCN2021107555-appb-000152
称取化合物38-149(0.2g，0.0443mmol)，加入二氯甲烷(20mL)，再加入TFA(2mL)，超声溶解，搅拌反应。反应结束，加甲基叔丁基醚与正己烷，抽滤，得产品0.14g，产率100％。
38-153
Figure PCTCN2021107555-appb-000153
称取化合物38-152(0.14g，0.1443mmol)，加入DMF(20mL)超声溶解，-5℃条件下搅拌20分钟，缓慢加入DIEA(0.29mL，1.772mol)，搅拌20分钟后加入M-SCM-10K(2.11g,0.1950mmol)，1小时后移至常温下避光低速搅拌7天，反应结束，加甲基叔丁基醚与正己烷，抽滤，滤饼干法上样，柱层析。用1％氨水+5％-25％甲醇/二氯甲烷梯度洗脱得产品1.5g，产率75％。
38-155
Figure PCTCN2021107555-appb-000154
称取38-153(1.3g,0.0288mmol)、1-(2-氨基乙基)-1H-吡咯-2,5-二酮盐酸盐(0.0808g,0.5767mmol，购于上海雷祺生物)、HBTU(0.2621g,0.6912mmol)、HOBT(0.0934g,0.6912mmol)投入500mL反应瓶中，加入DMF(10mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(0.3427mL，2.0736mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，蒸干得产品1.3克。
38-161
Figure PCTCN2021107555-appb-000155
称取38-155(1.3g,0.0276mmol)、PPT-iRGD(0.8847g,0.5527mmol,购于丹港生物科技)、投入250mL反应瓶中，加DMSO(50mL)溶液溶解，40℃条件下避光搅拌2天，常温搅拌3天。反应结束，在反应液中加甲基叔丁基醚，倾倒，固体不用10％甲醇/二氯甲烷溶解，加硅胶粉(5g)，干法上样，柱层析。用5％-10％甲醇/二氯甲烷洗脱得产品1.2g，产率57.14％。
1H-NMR(600MHz,DMSO-d 6)δ9.57–9.13(m,31H),9.09–8.63(m,10H),8.02–7.91(m,50H),7.91–7.80(m,75H),7.79–7.70(m,4H),7.69–7.54(m,2H),7.53–7-48(s,1H),7.45–7.38(s,1H),7.37–7.27(m,2H),7.26–7.19(s,1H),3.82–3.77(m,15H),3.76–3.72(m,13H),3.67–3.63(m,5H),3.62–3.52(m,63H),3.55–3.42(m,3934H),3.15–3.07(m,58H),3.06–3.01(m,44H),2.99–2.95(m,13H),2.94–2.85(m,625H),2.78–2.76(m,6H),2.75–2.70(m,155H),2.69–2.66(m,11H),2.63–2.59(m,4H),2.58–2.53(m,132H),2.52–2.45(m,134H),1.69–1.62(m,11H),1.59–1.49(m,8H),1.42–1.34(m,13H),1.31–1.22(m,460H),1.20–1.10(m,27H)。
38-192的合成：
Figure PCTCN2021107555-appb-000156
45-16
Figure PCTCN2021107555-appb-000157
称1.2-双(2-氨基乙氧基)乙烷(50mL,340.7327mmol，购于Tcl)投入500mL反应瓶中，加二氯甲烷(150mL)溶解，加三乙胺(94.9828mL,681.4654mmol)，在搅拌条件下缓慢分批加入(Boc) 2O(74.3751g,340.7327mmol)，常温搅拌过夜。次日在反应液中直接加入硅胶粉，蒸干，上样，柱层析。用2％-5％甲醇/二氯甲烷梯度洗脱得产品9.3g，产率10％。
38-172
Figure PCTCN2021107555-appb-000158
将45-16(8g，11.9225mmol)、6-马来酰亚胺基己酸(2.7701g，13.1147mmol)、HBTU(6.782g,17.8838mmol)、HOBT(2.4166g,17mmol)加入到装有500ml烧瓶中，加入适量DMF(50mL)使其溶解，置于-5摄氏度下避光搅拌30分钟，再缓慢滴加DIEA(8.8676mL，53.6512mmol)，在-5摄氏度下避光搅拌反应1小时后，移至室温下避光搅拌过夜。反应结束,用乙酸乙酯与饱和食盐水萃取处理，浓缩，蒸干有机相得到产品10g。
38-179
Figure PCTCN2021107555-appb-000159
将38-172(10g,22.649mmol)加入250mL烧瓶中，加入二氯甲烷(15mL)和TFA(25ml，339.7325mmol)使之溶解，反应在室温下避光搅拌过夜。反应结束，用旋转蒸发仪将反应液蒸至油状，再加入甲基叔丁基醚(60mL)，反应液中粉末状固体析出，抽滤，收集滤饼，然后用甲醇/二氯甲烷(1:4)溶液(100mL)溶解，加入硅胶粉(20g)，蒸干至粉末状，干法上样，柱层析，用3％-10％甲醇的二氯甲烷混合溶液梯度洗脱，收集浓缩，真空烘箱干燥，得到产品3g，产率38.96％。
38-191
Figure PCTCN2021107555-appb-000160
称取38-153(1g,0.0221mmol)、38-179(0.1508g,0.442mmol)、HBTU(0.2012g,0.5304mmol)、HOBT(0.0717g,1.5912mmol)投入500mL反应瓶中，加入DMF(10mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(0.2630mL，1.5912mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，蒸干得产品1克。
38-192
Figure PCTCN2021107555-appb-000161
称取38-191(1g,0.0199mmol)、PPT-iRGD(0.6370g,0.3979mmol)、投入250mL反应瓶中，加DMSO(50mL)溶液溶解，40℃条件下避光搅拌2天，常温搅拌3天。反应结束，在反应液中加甲基叔丁基醚，倾倒，固体不用10％甲醇/二氯甲烷溶解，加硅胶粉(5g)，干法上样，柱层析。用5％-10％甲醇/二氯甲烷洗脱得产品0.8g，产率50％。
1H-NMR(600MHz,DMSO-d 6)δ8.44-8.33(m,64H),8.25-8.18(m,47H),7.29-7.22(m,144H),4.90-4.79(m,8H),4.66-4.45(m,101H),4.38-4.19(m,194H),3.80-3.71(m,83H),3.55-3.45(m,3843H),3.20-3.04(m,208H),2.94-2.86(m,72H),2.83-2.72(m,153H),2.35-2.31(m,20H),2.06-2.02(m,29H),1.92-1.83(m,189H),1.68-1.42(m,435H),1.30-1.21(m,155H),0.88-0.83(m,192H)。
52-95的合成：
Figure PCTCN2021107555-appb-000162
Figure PCTCN2021107555-appb-000163
45-114
Figure PCTCN2021107555-appb-000164
称取Boc-L-Lys(Fmoc)-OH(20g,42.6857mmol)、Gly-OBn(14.4g,42.6857mmol)、HBTU(24g,64.0286mmol)、HOBT(8.65g,64.0286mmol)投入500mL反应瓶中，加入DMF(40mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(31.7mL，192.085mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，蒸干得产品26克。
45-115
Figure PCTCN2021107555-appb-000165
称取化合物45-114(26g，42.6857mmol)，加入二氯甲烷(20mL)，再加入TFA(14mL，190.2855mmol)，超声溶解，搅拌反应。反应结束，反应液浓缩，再用乙酸乙酯(100mL)溶解，加饱和碳酸氢钠溶液至水相显碱性，分离有机相，水相用乙酸乙酯萃取3次(50mL×3)，直至水相无产品，合并有机相，再用饱和食盐水清洗一次(50mL)，浓缩蒸干得产品22g。
45-124
Figure PCTCN2021107555-appb-000166
称取Boc-L-Lys(Boc)-OH(7.3g,21.334mmol)、45-115(11g,21.334mmol)、HBTU(12.1363g,32.0016mmol)、HOBT(4.3241g,32.0016mmol)投入500mL 反应瓶中，加入DMF(80mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(15mL，96.003mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，干法上样，柱层析，用50％-100％乙酸乙酯/石油醚梯度洗脱得产品15克，产率83％。
45-126
Figure PCTCN2021107555-appb-000167
称取化合物45-124(15g，17.7725mmol)，加入二氯甲烷(30mL)，再加入TFA(39mL，533.1754mmol)，超声溶解，搅拌反应。反应结束，反应液浓缩，再用乙酸乙酯(100mL)溶解，加饱和碳酸氢钠溶液至水相显碱性，分离有机相，水相用乙酸乙酯萃取3次(150mL×3)，直至水相无产品，合并有机相，再用饱和食盐水清洗一次(150mL)，浓缩蒸干得产品11.44g。
45-127
Figure PCTCN2021107555-appb-000168
称取Boc-L-Lys(Boc)-OH(13.022g,37.5911mmol)、45-126(11.4g,17.0868mmol)、HBTU(19.5538g,51.5604mmol)、HOBT(6.9668g,51.5604mmol)投入500mL反应瓶中，加入DMF(80mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(25.4mL，153.7812mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，干法上样，柱层析，用1％-55甲醇/二氯甲烷梯度洗脱得产品18克，产率81％。
45-135
Figure PCTCN2021107555-appb-000169
取反应物45-127(1.1g,0.8458mmol)置于250mL烧瓶中，加DMF溶液(50mL)，加入吗啉(1.4745mL,16.9155mmoL)，常温搅拌反应，3小时后，反应结束。饱和食盐水(150mL)和乙酸乙酯(200mL)，分离有机相，水相用乙酸乙酯萃取3次(150mL×3)，水相中无产品，合并有机相，用饱和食盐水清洗2次(150mL×2)，有机相蒸干得产品0.912克。
21-221
Figure PCTCN2021107555-appb-000170
称赤藓醇(4.5g,36.849mmol)投500mL两颈反应瓶中，加四氢呋喃(60mL)超声均匀，置于0℃条件下搅拌，通入氮气保护后，加叔丁醇钾(200mL,184.245mmol)，0℃下搅拌2小时后，加入溴乙酸苄酯(29.187mL,184.245mmol)，再搅拌3小时后移至常温下反应。反应结束，用纯水与乙酸乙酯萃取处理反应液，浓缩有机相，干法上样，柱层析。用1％-2％乙酸乙酯/石油醚梯度洗脱得产品5g，产率20％。
38-120
Figure PCTCN2021107555-appb-000171
称21-221(0.64g,0.9793mmol)与10％Pd/C(100mg)投入氢化反应釜中，缓慢加入DMF(30mL)搅拌溶解，通入氢气(300psi)，常温搅拌过夜。次日，用装有硅藻土的砂芯漏斗抽滤反应液，除去Pd/C，得产品的DMF溶液，直接投下一步反应。
52-86
Figure PCTCN2021107555-appb-000172
称取45-135(0.5g,0.46mmol)、HBTU(0.22g,0.576mmol)、HOBT(0.08g,0.576mmol)投入250mL反应瓶中，加入38-120(0.096mmol)的DMF溶液溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(0.3mL，1.728mmol)，-5℃条件下反应过夜。反应结束，在反应液中加纯水，抽滤，干法上样，柱层析。用2％甲醇/二氯甲烷洗脱得产品0.32g，产率50％。
52-88
Figure PCTCN2021107555-appb-000173
称52-86(0.32g,0.0694mmol)与10％Pd/C(75mg)投入氢化反应釜中，缓慢加入DMF(50mL)搅拌溶解，通入氢气(2MPa)，常温搅拌过夜。次日，用装有硅藻土的砂芯漏斗抽滤反应液，除去Pd/C，得产品的DMF溶液，直接投下一步反应。
52-90
Figure PCTCN2021107555-appb-000174
称取M-NH 2HCl-10K(3.065g,0.2914mmol)、HBTU(0.1579g,0.4164mmol)、HOBT(0.0562g,0..4164mmol)投入250mL反应瓶中，加入52-88(0.0667mmol)的DMF溶液溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(0.257mL，1.5545mmol)，-5℃条件下反应1小时后移至常温下低速避光反应。反应结束，在反应液中加甲基叔丁基醚，抽滤，干法上样，柱层析。用5％-6％甲醇/二氯甲烷洗脱得产品1.2g。
52-92
Figure PCTCN2021107555-appb-000175
称取化合物52-90(1.5g，0.0325mmol)，加入二氯甲烷(10mL)，再加入TFA(0.78mL，10.4mol)，超声溶解，搅拌反应。反应结束，在反应液中加甲醇(30mL)，加过量碳酸钾，搅拌30分钟，抽滤，烘干得产品2g。
52-93
Figure PCTCN2021107555-appb-000176
称取52-92(0.0325mmol)、6-马来酰亚胺基己酸(0.137g,0.65mmol)、HBTU(0.295g,0.78mmol)、HOBT(0.105g,0.78mmol)投入250mL反应瓶中，DMF溶液溶解，-5℃条件下避光搅拌30分钟后。缓慢滴加DIEA(0.4mL，2.34mmol)，-5℃条件下反应1小时后移至常温下避光反应。反应结束，在反应液中加甲基叔丁基醚倾倒多次得产品1.5g。
52-95
Figure PCTCN2021107555-appb-000177
称取52-93(0.0325mmol)、PEPTIDE-iRGD(1.04g,0.65mmol)、投入250mL反应瓶中，加DMSO(50mL)溶液溶解，40℃条件下避光搅拌2天，常温搅拌3天。反应结束，在反应液中加甲基叔丁基醚，倾倒，固体不用10％甲醇/二氯甲烷溶解，加硅胶粉(5g)，干法上样，柱层析。用5％-25％甲醇/二氯甲烷洗脱得产品0.3g，产率44％。MALDI-TOF MS测定的分子量范围为72500-74070。
56-89的合成：
Figure PCTCN2021107555-appb-000178
Figure PCTCN2021107555-appb-000179
45-114
Figure PCTCN2021107555-appb-000180
称取Boc-L-Lys(Fmoc)-OH(20g,42.6857mmol)、Gly-OBn(14.4g,42.6857mmol)、HBTU(24g,64.0286mmol)、HOBT(8.65g,64.0286mmol)投入500mL反应瓶中，加入DMF(40mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(31.7mL，192.085mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，蒸干得产品26克。
45-115
Figure PCTCN2021107555-appb-000181
称取化合物45-114(26g，42.6857mmol)，加入二氯甲烷(20mL)，再加入TFA(14mL，190.2855mmol)，超声溶解，搅拌反应。反应结束，反应液浓缩，再用乙酸乙酯(100mL)溶解，加饱和氯化钠溶液，分离有机相，水相用乙酸乙酯萃取3次(50mL×3)，直至水相无产品，合并有机相，再用饱和食盐水清洗一次(50mL)，浓缩蒸干得产品22g。
45-124
Figure PCTCN2021107555-appb-000182
称取Boc-L-Lys(Boc)-OH(7.3g,21.334mmol)、45-115(11g,21.334mmol)、HBTU(12.1363g,32.0016mmol)、HOBT(4.3241g,32.0016mmol)投入500mL反应瓶中，加入DMF(80mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(15mL，96.003mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，干法上样，柱层析，用50％-100％乙酸乙酯/石油醚梯度洗脱得产品15克，产率83％。
45-126
Figure PCTCN2021107555-appb-000183
称取化合物45-124(15g，17.7725mmol)，加入二氯甲烷(30mL)，再加入TFA(39mL，533.1754mmol)，超声溶解，搅拌反应。反应结束，反应液浓缩，再用乙酸乙酯(100mL)溶解，加饱和碳酸氢钠溶液至水相显碱性，分离有机相，水相用乙酸乙酯萃取3次(150mL×3)，直至水相无产品，合并有机相，再 用饱和食盐水清洗一次(150mL)，浓缩蒸干得产品11.44g。
45-127
Figure PCTCN2021107555-appb-000184
称取Boc-L-Lys(Boc)-OH(13.022g,37.5911mmol)、45-126(11.4g,17.0868mmol)、HBTU(19.5538g,51.5604mmol)、HOBT(6.9668g,51.5604mmol)投入500mL反应瓶中，加入DMF(80mL)溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(25.4mL，153.7812mmol)，-5℃条件下反应过夜。反应结束，用纯水与乙酸乙酯萃取处理反应，饱和食盐水清洗，浓缩有机相，干法上样，柱层析，用1％-55甲醇/二氯甲烷梯度洗脱得产品18克，产率81％。
45-135
Figure PCTCN2021107555-appb-000185
取反应物45-127(1.1g,0.8458mmol)置于250mL烧瓶中，加DMF溶液(50mL)，加入吗啉(1.4745mL,16.9155mmoL)，常温搅拌反应，3小时后，反应结束。饱和食盐水(150mL)和乙酸乙酯(200mL)，分离有机相，水相用乙酸乙酯萃取3次(150mL×3)，水相中无产品，合并有机相，用饱和食盐水清洗2次(150mL×2)，有机相蒸干得产品0.912克。
21-221
Figure PCTCN2021107555-appb-000186
称赤藓醇(4.5g,36.849mmol)投500mL两颈反应瓶中，加四氢呋喃(60mL)超声均匀，置于0℃条件下搅拌，通入氮气保护后，加叔丁醇钾(200mL,184.245mmol)，0℃下搅拌2小时后，加入溴乙酸苄酯(29.187mL,184.245mmol)，再搅拌3小时后移至常温下反应。反应结束，用纯水与乙酸乙酯萃取处理反应液，浓缩有机相，干法上样，柱层析。用1％-2％乙酸乙酯/石油醚梯度洗脱得产品5g，产率20％。
38-120
Figure PCTCN2021107555-appb-000187
称21-221(0.64g,0.9793mmol)与10％Pd/C(100mg)投入氢化反应釜中，缓慢加入DMF(30mL)搅拌溶解，通入氢气(300psi)，常温搅拌过夜。次日，用装有硅藻土的砂芯漏斗抽滤反应液，除去Pd/C，得产品的DMF溶液，直接投下一步反应。
52-86
Figure PCTCN2021107555-appb-000188
称取45-135(0.5g,0.46mmol)、HBTU(0.22g,0.576mmol)、HOBT(0.08g,0.576mmol)投入250mL反应瓶中，加入38-120(0.096mmol)的DMF溶液溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(0.3mL，1.728mmol)，-5℃条件下反应过夜。反应结束，在反应液中加纯水，抽滤，干法上样，柱层析。用2％甲醇/二氯甲烷洗脱得产品0.32g，产率50％。
52-88
Figure PCTCN2021107555-appb-000189
称52-86(0.32g,0.0694mmol)与10％Pd/C(75mg)投入氢化反应釜中，缓慢加入DMF(50mL)搅拌溶解，通入氢气(2MPa)，常温搅拌过夜。次日，用装有硅藻土的砂芯漏斗抽滤反应液，除去Pd/C，得产品的DMF溶液，直接投下一步反应。
52-90
Figure PCTCN2021107555-appb-000190
称取M-NH 2HCl-10K(3.065g,0.2914mmol)、HBTU(0.1579g,0.4164mmol)、HOBT(0.0562g,0..4164mmol)投入250mL反应瓶中，加入52-88(0.0667mmol)的DMF溶液溶解，-5℃条件下搅拌30分钟后。缓慢滴加DIEA(0.257mL，1.5545mmol)，-5℃条件下反应1小时后移至常温下低速避光反应。反应结束，在反应液中加甲基叔丁基醚，抽滤，干法上样，柱层析。用5％-6％甲醇/二氯甲烷洗脱得产品1.2g。
1H-NMR(400MHz,DMSO-d 6)δ7.98-7.88(m,14H),7.71-7.60(m,15H),7.58-7.47(m,16H),7.45-7.31(m,6H),5.76(s,28H),3.8-3.35(m,3810H),3.20-3.12(m,13H),2.91-2.80(m,10H),2.71-2.60(m,14H),2.33-2.30(m,13H),2.25-2.09(m,22H),1.36-1.28(m,51H),1.20-1.11(m,49H),1.05-1.00(m,96H),
52-92
Figure PCTCN2021107555-appb-000191
称取化合物52-90(1.5g，0.0325mmol)，加入二氯甲烷(10mL)，再加入TFA(0.78mL，10.4mol)，超声溶解，搅拌反应。反应结束，在反应液中加甲醇(30mL)，加过量碳酸钾，搅拌30分钟，抽滤，烘干得产品2g。
1H-NMR(400MHz,DMSO-d 6)δ8.25-8.02(m,12H),7.92-7.80(m,11H),7.71-7.65(m,2H),5.76-5.60(m,8H),5.58-5.50(m,23H),3.75-3.70(m,27H),3.60-3.56(m,35H),3.51-3.40(m,3810H),2.71-2.65(m,12H),2.60-2.55(m,16H),1.71-1.65(m,15H),1.60-1.55(m,28H),1.52-1.47(m,17H),1.16-1.00(m,18H)
56-86
Figure PCTCN2021107555-appb-000192
称取52-92(0.0325mmol)、N-乙酰基-L-半胱氨酸(0.046g,0.286mmol，购于伊诺凯)、HBTU(0.147g,0.39mmol)、HOBT(0.052g,0.0.39mmol)投入250mL反应瓶中，DMF溶液溶解，-5℃条件下避光搅拌30分钟后。缓慢滴加DIEA(0.193mL，1.17mmol)，-5℃条件下反应1小时后移至常温下避光反应。反应结束，在反应液中加甲基叔丁基醚倾倒多次得产品1.5g。
1H-NMR(400MHz,DMSO-d 6)δ8.03-8.00(m,56H),5.75-5.68(m,5H),4.45-4.37(m,26H),3.55-5.45(m,3810H),1.81-1.75(m,80H),1.51-1.45(m,26H),1.40-1.33(m,133H),0.85-0.83(m,12H),
56-89
Figure PCTCN2021107555-appb-000193
称取56-86(0.0325mmol)、MI-AH-PPT-iRGD(1.04g,0.65mmol，购于丹港生物)、投入250mL反应瓶中，加DMSO(50mL)溶液溶解，40℃条件下避光搅拌2天，常温搅拌3天。反应结束，在反应液中加甲基叔丁基醚，倾倒，固体不用10％甲醇/二氯甲烷溶解，加硅胶粉(5g)，干法上样，柱层析。用5％-25％甲醇/二氯甲烷洗脱得产品0.3g，产率44％。
1H-NMR(400MHz,DMSO-d 6)δ8.35-8.30(m,7H),8.24-8.20(m,14H),8.17-8.10(m,48H),8.07-8.01(m,32H),8.00-7.94(m,20H),7.86-7.80(m,24H),7.78-7.71(m,34H),7.69-7.51(m,31H),7.45-7.37(m,144H),7.31-7.22(m,28H),5.69-5.60(m,5H),5.07-4.95(m,14H),4.40-4.35(m,61H),4.23-4.06(m,78H),3.76-3.65(m,120H),3.55-3.40(m,3810H),2.82-2.76(m,35H),2.65-2.60(m,16H),2.11-2.05(m,25H),1.78-1.70(m,125H),1.31-1.28(m,57H),1.23-119(m,150H),1.17-1.14(m,146H),1.10-1.04(m,317H),0.93-0.84(m,41H),0.80-0.76(m,240H)
实施例2 聚乙二醇偶联药物增效剂对于抗癌药物在人乳腺癌MDA-MB-231细胞BALB/c裸鼠皮下移植瘤模型的抗肿瘤作用的增效试验
1、配制方法
供试品：
1)量取适量的抗癌药物(例如49-166)，加入适量的生理盐水，配制成合适浓度的溶液。
2)量取适量的待测聚乙二醇偶联药物增效剂(例如45-164)+抗癌药物(例如49-166)，加入适量的生理盐水，配制成合适浓度的溶液。
阴性对照：直接使用生理盐水。
配制好的供试品和对照品给药前于2～8℃或冰盒保存，给药剩余的供试品和对照品按医疗垃圾处理。
2、细胞及实验动物
人乳腺癌细胞MDA-MB-231：来源于中国医学科学院基础所细胞资源中心，培养条件为RPMI1640+10％FBS，37℃、5％CO 2培养。
动物种属&品系：BALB/C裸鼠
动物等级：SPF级
实验动物来源：北京维通利华实验动物技术有限公司
肿瘤接种时周龄：约4-5周。
肿瘤接种时体重：约15～18g。同性别动物的体重在平均体重的80％～120％之间。
性别与数量：雌性，订购96只小鼠，筛选成模动物48只用于最终实验，剩余动物转交兽医或安乐死。
动物饲养于独立通风系统(IVC)中，每笼最多饲养同组6只动物，SPF级动物房，环境条件控制在室温20～26℃，相对湿度40％～70％，光照12小时明暗交替。在检疫驯养期间和试验期间，每天提供合格的鼠料(生产单位：北京科澳协力饲料有限公司)，动物均自由摄食并自由饮水。
3、模型的建立
复苏MDA-MB-231细胞，进行细胞传代扩增，待扩增至足量细胞数时，收集处于对数生长期的细胞，准备进行细胞接种。
根据实际细胞数，调整细胞浓度在1×10 8个/mL，与基质胶(Matrigel Basement Membrance Matrix，BD公司)按1:1体积混合，得到浓度为5×10 7个/mL的细胞悬液，以0.2mL/只接种于96只小鼠右侧腋窝皮下。在接种后观察肿瘤生长情况，最终筛选得到肿瘤体积为77.30～292.27mm 3的48只成瘤动物用于试验。
4、动物分组和给药剂量
根据瘤体积和体重随机分为8组，其中，1组(阴性对照组，生理盐水)、6组(49-166，48mg/kg)、8组(49-166+45-164，48+176mg/kg)，每组6只。静脉注射，各组在D1、D4、D7、D10、D13、D18、D21、D24给药，D27动物实施安乐死。
5、实验结果
5.1一般临床观察
观察频率及时间：实验期间，所有动物每天进行两次大体观察。
观察内容：包括动物精神状态、行为活动、摄食情况等。
5.2体重
检测动物：所有动物
检测时间：接收后、接种前、分组(即首次给药前)(D1)、以后每周2次、安乐死前称取动物体重。动物发生意外死亡或濒死安乐死时也称重。
5.3瘤径的测量
检测动物：所有动物
检测时间：分组(即首次给药当天为D1)、首次给药后每周2次、安乐死前，游标卡尺测量并记录肿瘤长、短径，计算肿瘤体积。
按照以下公式计算肿瘤体积：
V＝1/2×长径×短径 2
试验期间每天2次观察动物一般临床症状，共进行10次体重和瘤径测量，安乐死后剥取肿瘤，称量肿瘤重量。计算瘤体积、相对肿瘤体积RTV、相对肿瘤增殖率T/C％、瘤重抑制率IR TW％。
6、结果分析与评价
统计分析：本试验使用统计学软件SPSS13.0对数据进行处理，计量资料以平均值±标准误来表示。具体分析过程如下：
用单因素方差分析(ANOVA)进行统计分析，如果ANOVA有统计学意义(P≤0.05)且方差齐性，用Tukey test进行组间比较分析，若方差不齐性，则用Dunnett’s T3 test进行组间比较分析。
6.1根据肿瘤体积进行疗效评价
按照以下公式计算相对肿瘤体积(RTV)和相对肿瘤增殖率T/C％：
RTV＝V t/V 0
V t：每天测量肿瘤得到的瘤体积
V 0：初始瘤体积(给药前)
T/C％＝给药组的RTV平均值/对照组的RTV平均值×100％
以给药组相对肿瘤增殖率T/C％≤40％且该组RTV与阴性对照组RTV相比P≤0.05为有效，有肿瘤增长抑制作用；反之，如果T/C％>40％，则为对肿瘤增长无抑制作用。
6.2根据肿瘤重量进行疗效评价
实验结束后，剥离肿瘤结节并进行称重，比较各组间肿瘤重量的差异以进一步计算肿瘤抑制率IR TW，以IR TW≥60％为有效性辅助参考指标，计算公式如下：
IR TW(％)＝(W 模型组-W 给药组)/W 模型组×100％
6.3大体解剖观察
对实验期间死亡动物以及观察期结束后安乐死动物进行大体解剖，对主要脏器进行观察，看是否有肉眼可见明显异常。
6.4照片记录
对安乐死动物及肿瘤进行照相。
具体结果如下：
整个试验过程中，各组动物未见明显异常。各组动物体重试验期间缓慢增长，各供试品组与第1组间未见显著性差异(P>0.05)。
阴性对照组(1组)整个试验中，肿瘤逐渐增长，至试验结束时(D27)，1组平均瘤体积为2962.92±2176.59mm 3，平均RTV为19.14±12.01；6、8组的平均肿瘤体积分别为1690.27±785.04mm 3、1273.37±358.45mm 3，平均RTV分别为13.03±5.43、10.55±5.73。
各组肿瘤增长趋势见图1。
至实验结束时(D27)，6、8组T/C％值分别为68.08％、55.10％，其IR TV％分别为31.92％、44.90％。
试验结束时，动物安乐死后称量瘤重，1、6、8组的平均瘤重分别为2.555±2.207g、0.990±0.399g、0.684±0.165g。6、8组的IR TW％分别为61.24％、73.21％。
各组肿瘤重量抑制率示意图见图2。
结论：在本实验条件下，供试品49-166在48mg/kg剂量下、49-166在48mg/kg剂量与45-164在176mg/kg剂量联合(49-166+45-164)，尾静脉注射给药对人乳腺癌MDA-MB-231细胞BALB/c裸鼠皮下移植瘤的生长均有明显的抑制作用，49-166+45-164的抑瘤作用显著优于49-166。
发明人通过以上实验和结果发现，本发明的聚乙二醇偶联药物增效剂可以作为抗癌药物对皮下移植瘤(例如，人乳腺癌MDA-MB-231细胞BALB/c裸鼠皮下移植瘤)的抗肿瘤作用的增效剂，且增效明显。

Claims (10)

  1. 式(I)所示的聚乙二醇偶联药物增效剂或其药学上可接受的盐，
    Figure PCTCN2021107555-appb-100001
    其中：
    M 1为
    Figure PCTCN2021107555-appb-100002
    PEG 1为单臂聚乙二醇链段，PEG 1通过羰基与L 1相连或者PEG 1的末端带有氨基或活化的氨基，PEG 1的数均分子量为5k-40k，优选为5k-10k或10k-40k，更优选为10k；
    L 1为
    Figure PCTCN2021107555-appb-100003
    各r 1独立地为1、2、3、4、5或6，优选为1、2、3或4，更优选为3或4；r 2为1、2、3、4、5或6，优选为1、2、3或4，更优选为1或2；
    优选地，L 1为
    Figure PCTCN2021107555-appb-100004
    V为
    Figure PCTCN2021107555-appb-100005
    Y1、Y0各自独立地选自
    Figure PCTCN2021107555-appb-100006
    Figure PCTCN2021107555-appb-100007
    r 1为1、2、3、4、5或6，r 1优选为1、2、3或4，r 1更优选为3或4，各r 2独立地为1、2、3、4、5或6，各r 2独立地优选为1、2、3或4，各r 2独立地更优选为1或2；
    优选地，Y1、Y0各自独立地选自
    Figure PCTCN2021107555-appb-100008
    或者优选地，Y1选自
    Figure PCTCN2021107555-appb-100009
    r 1为3或4，各r 2独立地为1或2；
    更优选地，Y1选自
    Figure PCTCN2021107555-appb-100010
    或者优选地，Y0选自
    Figure PCTCN2021107555-appb-100011
    r 1为3或4，r 2为1或2；
    更优选地，Y0选自
    Figure PCTCN2021107555-appb-100012
    P为-L V-T；
    L V选自
    Figure PCTCN2021107555-appb-100013
    Figure PCTCN2021107555-appb-100014
    各r 0独立地为1、2、3、4、5或6，各r 0独立地优选为3、4、5或6，各r 0独立地更优选为5或6，各r 2独立地为1、2、3、4、5或6，各r 2独立地优选为1、2、3或4，各r 2独立地更优选为1或2；
    优选地，L V选自
    Figure PCTCN2021107555-appb-100015
    Figure PCTCN2021107555-appb-100016
    T为
    Figure PCTCN2021107555-appb-100017
  2. 权利要求1的聚乙二醇偶联药物增效剂或其药学上可接受的盐，其中，所述聚乙二醇偶联药物增效剂选自：
    Figure PCTCN2021107555-appb-100018
    Figure PCTCN2021107555-appb-100019
    Figure PCTCN2021107555-appb-100020
    Figure PCTCN2021107555-appb-100021
    Figure PCTCN2021107555-appb-100022
    Figure PCTCN2021107555-appb-100023
  3. 制备权利要求1所述的聚乙二醇偶联药物增效剂或其药学上可接受的盐的方法，其包括：
    (1)制备中间体
    Figure PCTCN2021107555-appb-100024
    其中：
    M 1、L 1、Y 1、Y 0如权利要求1所限定，
    Pro 2为羧基的保护基，优选地，Pro 2为苄氧基，
    Pro 1为氨基或羧基的保护基，优选地，Pro 1为氨基的保护基时，Pro 1为叔丁氧羰基，优选地，Pro 1为羧基的保护基时，Pro 1为叔丁氧基；
    (2)将中间体
    Figure PCTCN2021107555-appb-100025
    进行第一脱保护，得到中间体
    Figure PCTCN2021107555-appb-100026
    其中，
    Figure PCTCN2021107555-appb-100027
    的末端带有羧基，；
    (3)使中间体
    Figure PCTCN2021107555-appb-100028
    与PEG 1发生酰胺化反应，得到 中间体
    Figure PCTCN2021107555-appb-100029
    其中，PEG 1如权利要求1所限定；
    (4)Pro 1为氨基的保护基时，将中间体
    Figure PCTCN2021107555-appb-100030
    进行第二脱保护，得到中间体
    Figure PCTCN2021107555-appb-100031
    其中，
    Figure PCTCN2021107555-appb-100032
    的末端带有氨基；
    或者，Pro 1为羧基的保护基时，将中间体
    Figure PCTCN2021107555-appb-100033
    进行第三脱保护，得到中间体
    Figure PCTCN2021107555-appb-100034
    其中，
    Figure PCTCN2021107555-appb-100035
    的末端带有羧基；
    (5)使中间体
    Figure PCTCN2021107555-appb-100036
    与
    Figure PCTCN2021107555-appb-100037
    发生酰胺化反应，得到中间体
    Figure PCTCN2021107555-appb-100038
    其中，r 0如权利要求1所限定；
    或者，使中间体
    Figure PCTCN2021107555-appb-100039
    与
    Figure PCTCN2021107555-appb-100040
    Figure PCTCN2021107555-appb-100041
    发生酰胺化反应，得到中间体
    Figure PCTCN2021107555-appb-100042
    Figure PCTCN2021107555-appb-100043
    其中，r 0、r 2如权利要求1所限定；
    (6)使中间体
    Figure PCTCN2021107555-appb-100044
    Figure PCTCN2021107555-appb-100045
    与
    Figure PCTCN2021107555-appb-100046
    发生加成反应，得到权利要求1所述的聚乙二醇偶联药物增效剂；
    优选地，中间体
    Figure PCTCN2021107555-appb-100047
    选自以下：
    Figure PCTCN2021107555-appb-100048
    Figure PCTCN2021107555-appb-100049
    Figure PCTCN2021107555-appb-100050
  4. 中间体，其用于制备权利要求1-2任一项所述的聚乙二醇偶联药物增效剂或其药学上可接受的盐，所述中间体选自以下：
    Figure PCTCN2021107555-appb-100051
    Figure PCTCN2021107555-appb-100052
    Figure PCTCN2021107555-appb-100053
  5. 组合物，其包含权利要求1-2任一项所述的聚乙二醇偶联药物增效剂或其药学上可接受的盐；任选地，所述组合物还包含一种或多种药学上可接受的辅料。
  6. 权利要求5的组合物，其中，所述组合物还包含抗癌药物。
  7. 权利要求6的组合物，其中，所述抗癌药物为式(A)所示的聚乙二醇偶联药物或其药学上可接受的盐，
    Figure PCTCN2021107555-appb-100054
    其中：
    M 2为
    Figure PCTCN2021107555-appb-100055
    j为3或4；
    PEG 2为单臂聚乙二醇链段，PEG 2通过羰基与L 2相连或者PEG 2的末端带有氨基或活化的氨基，PEG 2的数均分子量为5k-40k，优选为5k-10k或10k-40k，更优选为5k；
    L 2为
    Figure PCTCN2021107555-appb-100056
    各r 1独立地为1、2、3、4、5或6，各r 1独立地优选为1、2、3或4，各r 1独立地更优选为3或4,
    优选地，L 2为
    Figure PCTCN2021107555-appb-100057
    W为
    Figure PCTCN2021107555-appb-100058
    Z 2、Z 1、Z 0各自独立地为
    Figure PCTCN2021107555-appb-100059
    r 1为1、2、3、4、5或6，r 1优选为1、2、3或4，r 1更优选为3或4，各r 2独立地为1、2、3、4、5或6，各r 2独立地优选为1、2、3或4，各r 2独立地更优选为1或2，
    优选地，Z 2、Z 1、Z 0各自独立地为
    Figure PCTCN2021107555-appb-100060
    Figure PCTCN2021107555-appb-100061
    或者优选地，Z 2为
    Figure PCTCN2021107555-appb-100062
    r 1为3或4，
    更优选地，Z 2为
    Figure PCTCN2021107555-appb-100063
    或者优选地，Z 1为
    Figure PCTCN2021107555-appb-100064
    各r 2独立地为1或2，
    更优选地，Z 1为
    Figure PCTCN2021107555-appb-100065
    或者优选地，Z 0为
    Figure PCTCN2021107555-appb-100066
    r 2为1或2，
    更优选地，Z 0为
    Figure PCTCN2021107555-appb-100067
    Q为N-AC；
    N为
    Figure PCTCN2021107555-appb-100068
    或G，r 0为1、2、3、4、5或6，优选为3、4、5或6，更优选为5或6,
    优选地，N为
    Figure PCTCN2021107555-appb-100069
    或G；
    AC为SB7、NPB、SN38、LPT、PCB、DOX、PTX或AXT，优选为PTX或SN38。
  8. 权利要求7的组合物，其中，所述聚乙二醇偶联药物选自：
    Figure PCTCN2021107555-appb-100070
    Figure PCTCN2021107555-appb-100071
  9. 权利要求1-2任一项所述的聚乙二醇偶联药物增效剂或其药学上可接受的盐或者权利要求5所述的组合物在制备药物中的用途，所述药物用于增强治疗和/或预防疾病的疗效；
    优选地，所述疾病为癌症，所述癌症选自：结肠癌、白血病、淋巴瘤、膀胱癌、骨癌、脑瘤、髓母细胞瘤、胶质瘤、乳腺癌、腺瘤/类癌、肾上腺皮质癌、胰岛细胞癌、子宫颈癌、子宫内膜癌、卵巢癌、结肠直肠癌、皮肤癌、食管癌、眼癌、胆囊癌、胃癌、头颈癌、肝癌、黑色素瘤、卡波氏肉瘤、肾癌、口腔癌、肺癌、鼻咽癌、神经母细胞瘤、卵巢癌、胰腺癌、甲状腺癌、甲状旁腺阴茎癌、前列腺癌、尿道癌、阴道癌、外阴癌、肛门癌、肉瘤，以及所述癌症的转移。
  10. 权利要求6-8任一项所述的组合物在制备治疗和/或预防疾病的药物中的用途，所述疾病为权利要求6-8任一项所限定的抗癌药物所治疗的疾病；
    优选地，所述疾病为癌症，所述癌症选自：结肠癌、白血病、淋巴瘤、膀胱癌、骨癌、脑瘤、髓母细胞瘤、胶质瘤、乳腺癌、腺瘤/类癌、肾上腺皮质癌、胰岛细胞癌、子宫颈癌、子宫内膜癌、卵巢癌、结肠直肠癌、皮肤癌、食管癌、眼癌、胆囊癌、胃癌、头颈癌、肝癌、黑色素瘤、卡波氏肉瘤、肾癌、口腔癌、肺癌、鼻咽癌、神经母细胞瘤、卵巢癌、胰腺癌、甲状腺癌、甲状旁腺阴茎癌、前列腺癌、尿道癌、阴道癌、外阴癌、肛门癌、肉瘤，以及所述癌症的转移。
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