WO2021213492A1 - 一类载药的大分子及其制备方法 - Google Patents

一类载药的大分子及其制备方法 Download PDF

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WO2021213492A1
WO2021213492A1 PCT/CN2021/089174 CN2021089174W WO2021213492A1 WO 2021213492 A1 WO2021213492 A1 WO 2021213492A1 CN 2021089174 W CN2021089174 W CN 2021089174W WO 2021213492 A1 WO2021213492 A1 WO 2021213492A1
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group
dendrimer
drugs
active agent
linker
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French (fr)
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黄建
祝令建
管忠俊
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上海森辉医药有限公司
上海盛迪医药有限公司
江苏恒瑞医药股份有限公司
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Publication of WO2021213492A1 publication Critical patent/WO2021213492A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • 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
    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • 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
    • C08G73/00—Macromolecular compounds obtained by reactions forming a linkage containing nitrogen with or without oxygen or carbon in the main chain of the macromolecule, not provided for in groups C08G12/00 - C08G71/00
    • C08G73/02—Polyamines
    • 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
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present disclosure belongs to the field of medicine, and relates to dendrimers loaded with drugs and pharmacokinetic modifiers, and in particular, to linking drugs to dendrimers through specific linkers.
  • the reagent is covalently attached to the hydrophilic polymer backbone and the like.
  • these strategies can solubilize pharmaceutically active agents and in some cases improve bioavailability and targeting, there are difficulties in the release of pharmaceutically active agents.
  • the carrier will not reach the target organ before the drug molecule reaches the target. Will quickly degrade to release the active pharmaceutical agent.
  • the release rate of the pharmaceutically active agent from the carrier is variable, so that the drug cannot reach an effective therapeutic dose in the body or in the target organ.
  • Dendrimers have made significant progress in the fields of biotechnology and pharmaceutical applications (Xiangyang Shi et al. Sci China Mater, 2018, 61(11), 1387–1403.).
  • Dendrimers are specific types of polymers with densely branched structures. Starting from the core molecule, the structure is similar to dendritic macromolecules. Divide into two branches and repeat this process until it grows dense enough to grow like a sphere-like bush (V Gajbhiye et al. Journal of Pharmacy and Pharmacology, 2009, 61,989-1003.), which is characterized by being more dense than ordinary polymers. Higher concentration of functional groups/unit molecular volume.
  • dendrimers such as their high degree of branching, multivalence, spherical structure and well-defined molecular weight, make them promising new stents for drug delivery.
  • research on the design and synthesis of biocompatible dendrimers and their applications in many areas of biological sciences including drug delivery has also received increasing attention.
  • CN103796684A discloses macromolecules connecting drugs and dendrimers via diacid linkers, especially via saturated branched or straight chain diacids containing C 1 -C 10 interrupted by oxygen, nitrogen or sulfur atoms ⁇ Child connection.
  • the present disclosure provides a macromolecule comprising:
  • the first end group which is a pharmaceutically active agent containing a hydroxyl group, an amino group, a carboxyl group or a sulfhydryl group or its residue A;
  • the second end group which is a pharmacokinetic modifier
  • first end group is covalently connected to the surface amino group of the dendrimer through a linker -X 1 -LX 2 -, X 1 is the linker between the linker and the pharmaceutically active agent, and X 2 is the linker and The linking group of dendrimer, where:
  • X 1 and X 2 are selected from -C(O)- or not present, and L is a C 1-10 linear or branched alkylene group optionally interrupted by one or more oxygen, sulfur or nitrogen atoms; when When at least one of X 1 and X 2 is absent, the C 1-10 linear or branched alkylene group is optionally substituted by a substituent; when X 1 and X 2 are both -C(O)-, the The C 1-10 linear or branched alkylene group is further substituted by a substituent; the substituent is selected from deuterium, hydroxyl, C 3-7 cycloalkyl, C 3-7 cycloalkylene, alkyl, One of alkoxy, haloalkyl, haloalkoxy, halogen, nitro, cyano, acyl, thioether, sulfinyl, sulfonyl, -NR 1 R 2 , aryl, heteroaryl, and heterocyclic group Or
  • X 1 and X 2 are selected from -C(O)- or not present, and L is a C 1-10 straight or branched chain alkenylene group optionally interrupted by one or more oxygen, sulfur or nitrogen atoms Alkynyl, the alkenylene and alkynylene groups are optionally selected from deuterium, hydroxy, alkyl, cycloalkyl, alkoxy, haloalkyl, haloalkoxy, halogen, nitro, cyano, acyl, sulfur Is substituted by one or more groups of ether, sulfinyl, sulfonyl, -NR 1 R 2 , aryl, heteroaryl and heterocyclic group, or
  • X 1 is selected from -C(O)- or does not exist, X 2 does not exist, and L is connected with the nitrogen atom of the amino group on the surface of the dendrimer to form a nitrogen-containing heterocyclic structure
  • L 1 is a C 1-10 linear or branched alkylene, alkenylene or alkynylene group optionally interrupted by one or more oxygen, sulfur or nitrogen atoms, the alkylene, alkenylene or alkynylene group
  • the alkynyl group is optionally selected from deuterium, hydroxy, alkyl, cycloalkyl, cycloalkylene, alkoxy, haloalkyl, haloalkoxy, halogen, nitro, cyano, acyl, thioether, sulfene Acyl, sulfonyl, -NR 1 R 2 , aryl, heteroaryl and heterocyclic group substituted by one or more groups
  • X 3 is selected from CR 5 or
  • X 1 is selected from -C(O)- or does not exist, X 2 does not exist, and L is L 2 is a C 1-10 linear or branched alkylene, alkenylene or alkynylene group optionally interrupted by one or more oxygen, sulfur or nitrogen atoms, the alkylene, alkenylene or alkynylene group
  • the alkynyl group is optionally selected from deuterium, hydroxy, alkyl, cycloalkyl, alkoxy, haloalkyl, haloalkoxy, halogen, nitro, cyano, acyl, thioether, sulfinyl, sulfonyl, -NR 1 R 2 , aryl, heteroaryl and heterocyclic group substituted by one or more groups, n is selected from 1 or 2, wherein the nitrogen-containing heterocyclic end is connected to the dendrimer, and the L 2 end is through -C(O)- or directly connected to the pharmaceutically active agent;
  • R 1 and R 2 are each independently selected from hydrogen, hydroxyl, C 1-6 alkyl, cycloalkyl, and C 1-6 alkoxy.
  • X 1 and X 2 are both -C(O)-
  • L is a C 1-6 linear or branched alkylene group interrupted by one or more oxygen, sulfur or nitrogen atoms
  • the C 1-6 linear or branched alkylene groups are independently selected from deuterium, hydroxyl, C 3-7 cycloalkyl, C 3-7 cycloalkylene, C 1-6 alkoxy, C 1-6
  • X 1 and X 2 are both -C(O)-
  • L is a C 1-6 linear or branched alkylene group interrupted by one to three atoms independently selected from sulfur or nitrogen
  • the C 1-6 linear or branched alkylene group is independently selected from hydroxyl, C 3-7 cycloalkyl, C 3-7 cycloalkylene, C 1-6 alkoxy, C 1-6
  • the C 1-6 linear or branched alkylene group is substituted with one or more C 3-7 cycloalkylene groups.
  • the linker is -C(O)-X-C(O)-,
  • R a and R b are each independently selected from hydrogen, C 1-6 alkyl, or R a , R b and the carbon atom to which they are connected together form a C 3-7 cycloalkyl;
  • R c and R d are each independently selected from Hydrogen, C 1-6 alkyl, or R c , R d and the carbon atom to which they are connected together form a C 3-7 cycloalkyl group; provided that at least one of R a , R b and the carbon atom to which they are connected together form a C 3- 7 cycloalkyl or at least one R c , R d and the carbon atom to which they are connected together form a C 3-7 cycloalkyl;
  • X' is selected from -CH 2 -, -O-, -S- and -NR 3 -;
  • R 3 is hydrogen or C 1-6 alkyl
  • s and t are each independently selected from an integer of 1 to 4.
  • the same or different groups can be selected for each occurrence of Ra , Rb , Rc , and Rd.
  • X is Wherein s1 and t1 are each independently selected from an integer of 1 to 4.
  • X' is selected from -O-, -S-, -NH-, and -N(CH 3 )-.
  • X 1 is -C(O)-, which is connected to the pharmaceutically active agent or residue A thereof;
  • X 2 is -C(O)-, which is connected to the dendron
  • the surface amino groups of the polymer D are connected to form an amide bond.
  • the pharmaceutically active agent or residue A thereof contains a hydroxyl group and forms an ester bond with X1.
  • X 1 is -C(O)-, which is connected to the pharmaceutically active agent or its residue A;
  • X 2 is -C(O)-, and
  • the surface amino groups of the dendrimer D are connected to form an amide bond, and the structure is selected from the following:
  • X 1 is -C(O)-, which is connected to the pharmaceutically active agent or residue A thereof;
  • X 2 is -C(O)-, which is connected to the dendron
  • the surface amino groups of polymer D are connected to form an amide bond, and the structure is as follows:
  • X 1 is -C(O)-, which is connected to the pharmaceutically active agent or its residue A; X 2 is absent, and a part of the group of L forms a nitrogen
  • the heterocyclic ring is connected with the surface amino group of the dendrimer D to form a carbon-nitrogen bond.
  • X 1 is -C(O)-, which is connected to the pharmaceutically active agent or its residue A; X 2 is absent, and a part of L is formed
  • the nitrogen heterocycle is connected with the surface amino group of the dendrimer D to form a carbon-nitrogen bond, and the structure is selected from the following:
  • the present disclosure also provides a macromolecule, which includes:
  • the first end group which is a pharmaceutically active agent containing a hydroxyl group, an amino group, a carboxyl group or a sulfhydryl group or a residue thereof;
  • the second end group which is a pharmacokinetic modifier
  • the first end group is covalently connected to the surface amino group of the dendrimer through a triacid linker, one carbonyl group in the triacid linker is connected to the pharmaceutically active agent or its residue, and the other two carbonyl groups are connected to the dendrimer
  • the same surface amino group of the shaped polymer is connected and thus forms a nitrogen-containing heterocyclic ring with the linker moiety.
  • the triacid linker is connected to the pharmaceutically active agent or its residue A and the dendrimer D to form the following structure:
  • X 4 is selected from C and N
  • L 3 is a C 1-10 alkylene, alkenylene or alkynylene group optionally interrupted by an oxygen, sulfur or nitrogen atom
  • p is an integer selected from 0-5.
  • the triacid linker is connected to the pharmaceutically active agent or its residue A and the dendrimer D to form a structure selected from the following:
  • D is a dendrimer
  • A is a pharmaceutically active agent or a residue thereof.
  • the pharmaceutically active agent or its residue contains a hydroxyl group and forms an ester bond with the carbonyl group of the triacid linker.
  • the present disclosure also provides a macromolecule, which includes:
  • the first end group which is Eribulin, Trabectedin or Lurbinectedin;
  • the second end group which is a pharmacokinetic modifier
  • the first end group is covalently linked to the surface amino group of the dendrimer through a diacid linker, the diacid linker comprising C 1-10 optionally interrupted by one or more sulfur or nitrogen atoms Alkyl chain.
  • the C 1-10 alkyl chain contained in the diacid linker is not interrupted by oxygen, sulfur or nitrogen atoms.
  • the diacid linker can be selected from
  • the diacid linker comprises a C 1-10 alkyl group interrupted by one or more oxygen, sulfur or nitrogen atoms.
  • the diacid linker can be selected from
  • the present disclosure also provides a macromolecule, which includes:
  • the first end group which is a pharmaceutically active agent containing a hydroxyl group, an amino group, a carboxyl group, or a sulfhydryl group or a residue thereof, and is not docetaxel;
  • the second end group which is a pharmacokinetic modifier
  • first end group is covalently connected to the surface amino group of the dendrimer through a linker selected from:
  • selection of the linker of the present disclosure can provide a desired drug release rate, for example, fast release or slow release.
  • the release rate of the pharmaceutically active agent of the macromolecule is faster than that that does not depend on the delivery of the macromolecule, and may be at least 1 times faster. In some embodiments, the release rate of the pharmaceutically active agent of the macromolecule is slower than the release that does not depend on the delivery of the macromolecule, and may be two times, three times, four times, five times, six times, seven times, eight times slower. , Nine times, ten times, or ten times or more, fifteen times or more, twenty times or more, or thirty times or more. Macromolecules with low release rates are suitable for formulating macromolecules into drugs that are slowly released within a period of 1 week to 3 months, 1 month to 6 months, or more than 6 months.
  • the rapid release is preferably within 0-8 hours, especially within 0-4 hours, especially within 0-2 hours, and more particularly within 5-60 minutes, to release more than 50% of the pharmaceutically active agent.
  • Moderate release preferably releases more than 50% of the pharmaceutically active agent within 1-72 hours, especially within 2-48 hours.
  • the release rate of the pharmaceutically active agent can be controlled by selecting a suitable linker, and the release rate also depends on the characteristics of the pharmaceutically active agent.
  • the pharmaceutically active agent is linked to the dendrimer through the same linker.
  • the pharmaceutically active agent is connected to the dendrimer through two or more linkers, so that the pharmaceutically active agent can be released from the macromolecule at different release rates.
  • the first end group and the second end group are present in a ratio of 1:2-2:1, especially 1:2, 1:1, 2:1.
  • the macromolecule includes a third end group that is a blocking group, a drug, or a targeting group.
  • the blocking group may be an acyl group.
  • the ratio of the first end group, the second end group, and the third end group is 1:1:1-1:2:2, especially 1:2:1.
  • at least 50% of the end groups include one of the first end group or the second end group.
  • the pharmaceutically active agent binds to greater than 14%, 25%, 27%, 30%, 39%, 44%, or 48% of surface amino groups.
  • the pharmacokinetic modifier binds to greater than 15%, 25%, 30%, 33%, or 46% of the total number of surface amino groups.
  • the pharmaceutically active agent of the present disclosure may be selected from: anesthetics, antacids, antibodies, anti-infectives, biological products, cardiovascular drugs, contrast agents, diuretics, blood tonics, immunosuppressants, hormones and the like, nutritional products , Ophthalmic drugs, pain treatment agents, respiratory drugs, adjuvants, anabolic agents, antiarthritis drugs, anticonvulsants, antihistamines, anti-inflammatory drugs, antiulcer drugs, behavior modification drugs, tumor drugs, central nervous system drugs, Contraceptives, diabetes treatment drugs, fertility drugs, growth promoters, hemostatic drugs, immunostimulants, muscle relaxants, obesity treatments, osteoporosis drugs, peptides, tranquilizers and tranquilizers, urethral acidifiers or vitamins.
  • the pharmaceutically active agent is a tumor drug, steroids, opioid analgesics, respiratory system drugs, central nervous system (CNS) drugs, hypercholesterolemia drugs, antihypertensive drugs, antibiotics, immunosuppressive drugs, Antibacterial agents, luteinizing hormone releasing hormone (LHRH) agonists, LHRH antagonists, antiviral drugs, antiretroviral drugs, estrogen receptor modulators, somatostatin analogs, anti-inflammatory drugs, vitamin D2 analogs, Synthetic thyroxine, antihistamine, antifungal or non-steroidal anti-inflammatory drug (NSAID).
  • NSAID non-steroidal anti-inflammatory drug
  • the tumor drugs include taxanes (such as paclitaxel, cabazitaxel, and docetaxel), camptothecin and its analogs (such as irinotecan and topotecan), and other anti-tumor drugs.
  • taxanes such as paclitaxel, cabazitaxel, and docetaxel
  • camptothecin and its analogs such as irinotecan and topotecan
  • other anti-tumor drugs include taxanes (such as paclitaxel, cabazitaxel, and docetaxel), camptothecin and its analogs (such as irinotecan and topotecan), and other anti-tumor drugs.
  • Microtubule agents such as vinflunine
  • nucleosides such as gemcitabine, cladribine, fludarabine, capecitabine, decitabine, azacitidine, clofarabine and nelarabine
  • Kinase inhibitors such as dasatinib (sprycel), temisirolimus, dasatinib, AZD6244, AZD1152, PI-103, R-roscovitine, oromocin, and purvalanol A
  • Eberberin B analogs such as ixabepilone
  • anthrocyclines such as amrubicin, doxorubicin, epirubicin and valrubicin
  • ascidian derivatives such as trobe Trabectecin, lurbinectedin
  • proteasome inhibitors such as bortezomib
  • other topoisomerase inhibitors such as intercalators and alkylating agents
  • microtubule inhibitors such
  • the pharmaceutically active agent is selected from taxanes, camptothecin derivatives, nucleoside drugs, anthracyclines, ascidian derivatives, proteasome inhibitors, microtubule inhibitors , BCL-2 inhibitors, BCL-X L inhibitors, selective nuclear export inhibitors, antimetabolites, tyrosine kinase inhibitors, PLK1 inhibitors, CDK4/6 inhibitors, BTK inhibitors, non-steroidal hormones Receptor antagonists and steroids, preferably taxanes, BCL-2 inhibitors and BCL- XL inhibitors.
  • the steroids include synthetic steroids (such as testosterone, dihydrotestosterone, and ethinyl estradiol) and corticosteroids (such as cortisone, prednisilone, budesonide, triamcinolone) , Fluticasone, Mometasone, Amcinonide, Flucinolone, Fluocinanide, Desonide, Clofloxasone, Prednisolone, Fluocorone, Dexamethasone, Betamethasone and Fluoride Prednidine (fluprednidine)).
  • synthetic steroids such as testosterone, dihydrotestosterone, and ethinyl estradiol
  • corticosteroids such as cortisone, prednisilone, budesonide, triamcinolone
  • Fluticasone Mometasone
  • Amcinonide Flucinolone
  • Fluocinanide Desonide
  • Clofloxasone Prednisolone
  • Fluocorone Dex
  • the opioid analgesics include morphine, oxymorphone, naloxone, codeine, oxycodone, methylnaltrexone, hydromorphone, buprenorphine, and eto coffee.
  • the respiratory system drugs include bronchodilators, inhaled steroids and decongestants, especially salbutamol, ipratropium bromide, montelukast and formoterol.
  • the CNS drugs include antipsychotics (such as quetiapine) and antidepressants (such as venlafaxine).
  • antipsychotics such as quetiapine
  • antidepressants such as venlafaxine
  • the hypercholesterolemia drugs include ezetimibe and statins such as simvastatin, lovastatin, atorvastatin, fluvastatin, pitavastatin, pravastatin, and pravastatin. Suvastatin.
  • the antihypertensive drugs include losartan, olmesartan, medoxomil, metrolol, travoprost, and bosentan.
  • the immunosuppressive drugs include glucocorticoids, cytostatics, antibody fragments, anti-immunophiles, interferons, TNF binding proteins, especially calcineurin inhibitors (cacineurin inhibitors) such as taco Moss, mycophenolic acid and its derivatives, such as mycophenolate mofetil and cyclosporine.
  • calcineurin inhibitors cacineurin inhibitors
  • the antibacterial agent includes antibiotics, such as amoxicillin, meropenem, and clavulanic acid.
  • the LHRH agonist includes goserelin acetate, deslorelin and leuprolide.
  • the LHRH antagonist includes cetrorelix, ganirelix, abarelix, and degarelix.
  • the antiviral drugs include nucleoside analogs such as lamivudine, zidovudine, abacavir and entecavir, and antiretroviral drugs include protease inhibitors such as atazanavir , Lapinavir and ritonavir.
  • the estrogen receptor modulator includes raloxifene and fulvestrant.
  • the somatostatin analogue includes octreotide.
  • the anti-inflammatory drugs include mesalazine and suitable NSAIDs include paracetamol (acetaminophen).
  • the vitamin D2 analog includes paricalcitol.
  • the synthetic thyroxine includes levothyroxine.
  • the antihistamine includes fexofenadine.
  • the antifungal agent includes azoles such as viriconazole.
  • the pharmaceutically active agent is Eribulin. In some embodiments, the pharmaceutically active agent is docetaxel. In some embodiments, the pharmaceutically active agent is Trabectidine. In some embodiments, the pharmaceutically active agent is lurbinectedin.
  • the pharmaceutically active agent is selected from the group consisting of docetaxel, irinotecan, gemcitabine, capecitabine, decitabine, azacitidine, doxorubicin, epirubicin, Trabectidine, rubetidine, bortezomib, eribulin, celinisol, venetoc, tecitaxel, pemetrexed, cabazitaxel, cabozantinib, ancestral (Onvansertib), compound 2 of the following formula, or structural modifications of these drug molecules,
  • the pharmaceutically active agent is slightly soluble or insoluble in an aqueous solution.
  • the second end group is a pharmacokinetic modifier, which can modify or mediate the pharmacokinetic characteristics of the pharmaceutically active agent or macromolecule, including absorption, distribution, metabolism and/or excretion.
  • the pharmacokinetic modifier prolongs the plasma half-life of the pharmaceutically active agent, so that the half-life of the pharmaceutically active agent attached to the macromolecule is longer than that of the pharmaceutically active agent alone or on a non-dendritic polymer carrier.
  • the half-life of the agent is longer.
  • the half-life of the macromolecule or composition is at least 2 times longer than the half-life of the pharmaceutically active agent alone or on a non-dendritic polymer carrier, and more preferably at least 10 times.
  • the pharmacokinetic modifier can be selected from polyethylene glycol, polyethyloxazoline, polyvinylpyrrolidone, polypropylene glycol, folate or folate derivatives of ligands for cell surface receptors .
  • the pharmacokinetic modifier is polyethylene glycol.
  • the polyethylene glycol has a molecular weight in the range of 220 to 5500 Da, for example, it can be 220-2500 Da, 570-2500 Da, 220-1100 Da, 570-1100 Da, 1000-5500 Da, 1000-2500 Da, 1000 -2300Da molecular weight.
  • the pharmacokinetic modifier forms an amide bond with an amino group on the surface of the dendrimer.
  • Targeting groups are reagents that bind to biological target cells, organs or tissues with some selectivity, thereby helping to direct macromolecules to specific targets in the body and allow them to accumulate in the target cells, organs or tissues.
  • the targeting group can provide a mechanism for macromolecules that actively enter cells or tissues through receptor-mediated endocytosis.
  • Specific examples include lectins and antibodies and other ligands (including small molecules) that target cell surface receptors. The interaction can be through any type of bond or association (including covalent bonds, ionic bonds and hydrogen bonds, van der Waals forces).
  • Suitable targeting groups include those that bind to cell surface receptors, for example, folate receptor, adrenergic receptor, growth hormone, luteinizing hormone receptor, estrogen receptor, epidermal growth factor receptor, fibroblasts Cell growth factor receptors (such as FGFR2), IL-2 receptors, CFTR and vascular epithelial growth factor (VEGF) receptors.
  • folate receptor adrenergic receptor
  • growth hormone luteinizing hormone receptor
  • estrogen receptor epidermal growth factor receptor
  • fibroblasts Cell growth factor receptors (such as FGFR2)
  • IL-2 receptors such as FGFR2
  • CFTR vascular epithelial growth factor
  • the targeting group is Luteinizing Hormone Releasing Hormone (LHRH) or a derivative thereof that binds to the Luteinizing Hormone Releasing Hormone Receptor.
  • LHRH Luteinizing Hormone Releasing Hormone
  • the targeting group is LYP-1, which is a peptide that targets the lymphatic system of the tumor but not the lymphatic system of the normal tissue.
  • the targeting group may be an RGD peptide.
  • the RGD peptide is a peptide containing the sequence -Arg-Gly-Asp-, which is the main integrin recognition site in the extracellular matrix protein.
  • the targeting group may be folic acid. Estrogen can also be used for target cells expressing estrogen receptors.
  • the targeting group can be attached to the dendrimer core directly or preferably via a linking group.
  • the linking group may be any divalent group capable of binding to the functional group on the core and the functional group on the targeting group.
  • the macromolecules of the present disclosure comprise dendrimers in which the outermost generation of structural units has surface amino groups.
  • the properties of the macromolecular dendrimer are not particularly important, provided that it has surface amino groups.
  • the dendrimer may be a polylysine, polylysine analog, polyamidoamine (PAMAM), polyethyleneimine (PEI), or polyether hydroxylamine (PEHAM) dendrimer.
  • PAMAM polyamidoamine
  • PEI polyethyleneimine
  • PEHAM polyether hydroxylamine
  • the dendrimer is polylysine or a polylysine analog.
  • Polylysine or polylysine analogues include core and 2-7 generations of lysine or lysine analogues, for example, 2nd, 3rd, 4th, 5th, 6th or 7th generation lysine Or lysine analogues.
  • the lysine has the structure shown in 1:
  • the lysine analogue has the structure shown in 2:
  • the lysine analogue has the structure shown in 3: Wherein a is an integer of 1 or 2; b and c are the same or different and are an integer of 1 to 4.
  • the lysine analogue has the structure shown in 4: Wherein a is an integer from 0 to 2; b and c are the same or different and are an integer from 2 to 6.
  • the lysine analogue has the structure shown in 5: Wherein a is an integer from 0 to 5; b and c are the same or different and are an integer from 1 to 5.
  • the lysine analogue has the structure shown in 6: Wherein a is an integer from 0 to 5; b and c are the same or different and are an integer from 0 to 5.
  • the lysine analogue has the structure shown in 7: Wherein a is an integer from 0 to 5; b and c are the same or different and are an integer from 1 to 5.
  • the lysine analogue has the structure shown in 8: Wherein a is an integer from 0 to 5; b, c and d are the same or different and are an integer from 1 to 5.
  • the lysine analogue has the structure shown in 9: Wherein a is an integer from 0 to 5; b and c are the same or different and are an integer from 1 to 5.
  • the dendrimer described in the present disclosure especially the core of polylysine or polylysine analogues can be selected from benzhydrylamine (BHA), benzhydrylamine of lysine (BHALys) or The lysine analogue or the structure shown in 10-26 below:
  • BHA benzhydrylamine
  • BHALys benzhydrylamine of lysine
  • 10-26 The lysine analogue or the structure shown in 10-26 below:
  • a is 1 to 9, preferably an integer of 1 to 5;
  • a, b and c are the same or different and are an integer of 1-5, and d is an integer of 0-100, preferably an integer of 1-30;
  • a and b are the same or different, and are an integer from 0 to 5;
  • a and c are the same or different and are an integer of 1-6, and b is an integer of 0-6;
  • a and d are the same or different and are an integer of 1-6, and b and c are the same or different and are an integer of 0-6;
  • a and b are the same or different and are an integer of 1-5, especially 1-3, especially 1;
  • a, b and c are the same or different, and are integers selected from 1-6;
  • a, b and c are the same or different, and are integers selected from 0-6;
  • a, b and c are the same or different, and are integers selected from 0-6;
  • a, b, and c are the same or different, and are integers from 0 to 6, and d, e, and f are the same or different, and are integers from 1-6;
  • a, b and c are the same or different and are integers of 1-6;
  • a, b, c and d may be the same or different, and are an integer of 0-6;
  • a, b, c and d may be the same or different, and are an integer of 1-6; or
  • a, b, c, and d may be the same or different, and are integers from 0-6, and e, f, g, and h may be the same or different, and are integers from 1-6.
  • the present disclosure also relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the macromolecule of the present disclosure and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition does not contain solubilizing excipients, such as polyethoxy castor oil, polysorbate.
  • the pharmaceutical composition is administered by transdermal, oral, injection and other methods.
  • the macromolecules of the present disclosure can be formulated into compositions that include compositions suitable for oral, rectal, topical, intranasal, inhalation, aerosol, ocular, or parenteral (including intraperitoneal, intravenous, subcutaneous or intramuscular) Internal injection).
  • the composition can conveniently be presented in unit dosage form and can be prepared by any method well known in the pharmaceutical art.
  • the method includes the step of associating the macromolecule with the carrier constituting one or more accessory ingredients.
  • the composition is prepared by associating the macromolecule with a liquid carrier to form a solution or suspension, or alternatively the macromolecule is associated with formulation components suitable for forming a solid, optionally particulate product, and then, if necessary, Shape the product into the desired delivery form.
  • the solid preparation of the present invention when it is a particle, will usually contain a particle size range of about 1 nanometer to about 500 micrometers. Generally, for solid formulations intended for intravenous administration, the particle diameter will generally range from about 1 nm to about 10 microns.
  • the composition may contain the macromolecules of the present invention, the macromolecules having a particle diameter of less than 1000nm, for example 5 to 1000nm, especially 5 to 500nm, especially 5 to 400nm (such as 5 to 50nm and especially 5 to 20nm) Of nanoparticles. In certain embodiments, the composition contains macromolecules having an average size of 5 to 20 nm.
  • the macromolecules are polydisperse in the composition, and the PDI is between 1.01 and 1.8, especially between 1.01 and 1.5, and especially between 1.01 and 1.2.
  • the macromolecules are monodisperse in the composition.
  • Particularly preferred are sterile, lyophilized compositions which are reconstituted in an aqueous vehicle before injection.
  • the composition contains macromolecules having an average size of 5 to 20 nm.
  • the particle size D 90 or D 50 of the macromolecule is less than 1000 nm, for example 5 to 1000 nm, especially 5 to 500 nm, especially 5 to 400 nm (such as 5 to 50 nm, especially 5 to 20 nm).
  • the composition contains molecules having a D 50 of 5 to 20nm.
  • the macromolecules of the present disclosure can also be used to provide controlled release and/or sustained release formulations of pharmaceutically active agents.
  • sustained-release formulations the ingredients of the formulation are selected to release macromolecules from the formulation over an extended period of time (such as days, weeks, or months).
  • Such formulations include transdermal patches or in implantable devices that can be deposited subcutaneously or by intravenous injection, subcutaneous injection, intramuscular injection, intradural injection or intracranial injection.
  • the diacid linker is selected to release most of its pharmaceutically active agent in a given time window.
  • the linker can be selected to release most of its pharmaceutically active agent after the accumulation time has passed. This may allow the delivery of high drug loads at the site where its effect is needed at a given point in time.
  • the linker is selected to release the pharmaceutically active agent at therapeutic levels for an extended period of time.
  • the formulation may have a variety of controlled release characteristics.
  • the formulation contains macromolecules in which the drugs are linked by different linkers, which allows for a burst of rapid release drugs and then slower release at a lower but constant therapeutic level over an extended period of time.
  • the formulation may have sustained release and controlled release characteristics.
  • composition of the formulation can be selected to release macromolecules over an extended period of time and the linker can be selected to deliver a constant low therapeutic level of the pharmaceutically active agent.
  • the pharmaceutically active agent is linked to the same molecule through different linkers.
  • each drug-linker combination is linked to a different macromolecule in the same formulation.
  • the macromolecule in the pharmaceutical composition, is formulated to release more than 50% of the pharmaceutically active agent within 5 minutes to 60 minutes. In some embodiments, in the pharmaceutical composition, the macromolecule is formulated to release more than 50% of the pharmaceutically active agent between 2 hours and 48 hours. In some embodiments, in the pharmaceutical composition, the macromolecule is formulated to release more than 50% of the pharmaceutically active agent between 5 days and 30 days.
  • Another aspect of the present disclosure provides a method for treating or inhibiting tumor growth, which comprises administering an effective amount of the macromolecule of the present invention or a pharmaceutical composition containing the same, wherein the pharmaceutically active agent as the first end group is a tumor drug.
  • the tumor described in the present disclosure is selected from melanoma, brain tumor, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, lung cancer, kidney cancer, breast cancer, ovarian cancer, prostate cancer, skin cancer, neuroblastoma , Sarcoma, osteochondroma, osteoma, osteosarcoma, seminoma, testicular tumor, uterine cancer, head and neck tumor, multiple myeloma, malignant lymphoma, polycythemia vera, leukemia, thyroid tumor, ureteral tumor, bladder Carcinoma, gallbladder cancer, cholangiocarcinoma, choriocarcinoma, and pediatric tumors (Ewing familial sarcoma
  • a method for reducing allergies after treatment with tumor drugs which comprises administering the macromolecule of the present disclosure or a pharmaceutical composition containing the same, wherein the composition is substantially free of solubilizing excipients such as Cremophor EL and Polysorbate 80.
  • a method for reducing the toxicity of a tumor drug or a preparation of a tumor drug which comprises administering the macromolecule of the present disclosure or a pharmaceutical composition containing the same, wherein the tumor drug is the first end of the macromolecule base.
  • the reduced toxicity is blood toxicity, neurotoxicity, gastrointestinal toxicity, cardiotoxicity, hepatotoxicity, nephrotoxicity, ototoxicity, or cerebral virality.
  • a method for reducing side effects associated with tumor drugs or preparations of tumor drugs which comprises administering the macromolecules of the present disclosure or pharmaceutical compositions containing the same, wherein the tumor drugs are the first of the macromolecules.
  • One end base is provided.
  • the reduced side effects are selected from the group consisting of neutropenia, leukopenia, thrombocytopenia, bone marrow toxicity, bone marrow suppression, neuropathy, fatigue, non-specific neurocognitive problems, dizziness, encephalopathy, anemia, dysgeusia, Difficulty breathing, constipation, anorexia, nail disorders, fluid retention, weakness, pain, nausea, vomiting, mucositis, alopecia, skin reactions, myalgia, allergies, and anaphylaxis.
  • the macromolecules or pharmaceutical compositions containing macromolecules of the present disclosure can reduce or eliminate the need for preoperative medication with agents such as corticosteroids and antihistamines.
  • dendrimers of macromolecules can be prepared by a divergent method or a convergent method or a mixture thereof.
  • each generation of the structural unit is sequentially added to the core or previous generation.
  • Protect surface generations with one or two surface amino groups. If one of the amino groups is protected, then the free amino group reacts with one of the linker, the linker-pharmacologically active agent, or the pharmacokinetic modifier. If both amino groups are protected and they are protected with different protecting groups, then one protecting group can be removed without removing the other protecting group. Remove one of the amino protecting groups and react with one of the linker, linker-pharmacologically active agent, or pharmacokinetic modifier. Once the initial end group has been attached to the dendrimer, the other amino protecting group is removed and another first and second end group are added. These groups are attached to surface amino groups through amide formation known in the art.
  • each generation of structural units is built on the previous generation to form dendrons.
  • the first and second end groups can be attached to the surface amino groups as described above.
  • each generation of the structural unit is added to the core or the previous generation of the structural unit.
  • the surface amino groups and end groups eg, first and second end groups, first and third end groups, or second and third end groups
  • the functionalized last generation is then added to the subsurface layer of the structural unit and the dendron is attached to the core.
  • the pharmaceutically active agent reacts with one of the carboxylic acids of the linker through an esterification reaction known in the art.
  • an activated carboxylic acid is formed such as using an acid chloride or an acid anhydride, and reacts with the hydroxyl group of the pharmaceutically active agent. If the pharmaceutically active agent has more than one hydroxyl group, the other hydroxyl groups can be protected.
  • the functional group on the core can be protected during the formation of the dendrimer and then deprotected and reacted with the targeting agent, linking group or targeting agent-linking group.
  • the core can be reacted with a linking group or targeting agent-linking group before forming the dendrimer.
  • the present disclosure also includes various deuterated forms of the macromolecule or a pharmaceutically acceptable salt thereof, and each available hydrogen atom in the macromolecule can be independently replaced by a deuterium atom.
  • each available hydrogen atom in the macromolecule can be independently replaced by a deuterium atom.
  • Those skilled in the art will know how to synthesize the deuterated form of the macromolecule of the present disclosure or a pharmaceutically acceptable salt thereof.
  • the present disclosure also includes isotope-labeled macromolecules in which one or more atoms in the macromolecule are replaced by atoms having a different atomic mass or mass number than the most common atomic mass or mass number in nature.
  • isotopes of macromolecules that can be used in the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, iodine, and chlorine, such as 3 H, 11 C, 14 C, 18 F, 123 I, or 125 I.
  • the macromolecules of the present disclosure may be in the form of pharmaceutically acceptable salts.
  • non-pharmaceutically acceptable salts also fall within the scope of the present disclosure, as these may be used as intermediates in the preparation of pharmaceutically acceptable salts or may be useful during storage or transportation.
  • Suitable pharmaceutically acceptable salts include, but are not limited to, salts of pharmaceutically acceptable inorganic acids (such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, sulfamic acid and hydrobromic acid), or pharmaceutically acceptable salts Organic acids (such as acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, maleic acid, citric acid, lactic acid, mucic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, Phenylacetic acid, methanesulfonic acid, toluenesulphonic, benzenesulfonic acid, salicylic acid, sulphanilic, aspartic acid, glutamic acid, ethylenediaminetetraacetic acid, stearic acid, palmitic acid, Oleic acid, lauric acid, pantothenic acid, tannic
  • interrupted by oxygen, sulfur or nitrogen atom(s) refers to oxygen
  • Sulfur or nitrogen atoms can be located inside the alkyl chain or at the end of the alkyl chain.
  • X 1 does not exist
  • X 2 is -C(O)-, which is connected with the surface amino group of the dendrimer D to form an amide bond
  • Discontinued with sulfur sulfur is located inside the alkyl chain, and oxygen is at the end of the alkyl chain.
  • alkyl refers to a linear or branched saturated hydrocarbon group having 1 to 10 carbon atoms. Where appropriate, the alkyl group may have a specified number of carbon atoms, for example, a C 1-4 alkyl group, and the alkyl group includes those having 1, 2, 3, or 4 carbon atoms in a linear or branched arrangement. alkyl.
  • alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methyl Butyl, 4-methylbutyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 5-methylpentyl, 2-ethylbutyl, 3 -Ethylbutyl, heptyl, octyl, nonyl and decyl.
  • alkylene refers to a straight or branched chain divalent alkyl group having 1 to 10 carbon atoms.
  • alkenyl is meant to include branched and straight chain olefins having 2 to 12 carbon atoms or olefins containing aliphatic hydrocarbon groups.
  • C 2-6 alkenyl means an alkenyl group having 2, 3, 4, 5, or 6 carbon atoms.
  • alkenyl groups include, but are not limited to, vinyl, allyl, 1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methylbut-2-enyl, 3-methylbut-1-enyl, 1-pentenyl, 3-pentenyl and 4-hexenyl.
  • alkynyl refers to olefins that include branched and straight-chain alkynyl groups having 2 to 12 carbon atoms or aliphatic hydrocarbon groups, or if the number of carbon atoms is specified, it means that specific number. For example, ethynyl, propynyl (e.g., 1-propynyl, 2-propynyl), 3-butynyl, pentynyl, hexynyl, and 1-methylpent-2-ynyl.
  • alkenylene and alkynylene refer to a partially unsaturated branched or straight chain divalent hydrocarbon group derived from an alkenyl or alkynyl group. In certain embodiments, such alkenylene groups are optionally substituted.
  • alkenylene groups include vinylene, propenylene, butenylene, pentenylene, hexenylene, heptenylene, octenylene, nonenylene, decenylene And similar groups; non-limiting examples of alkynylene groups include ethynylene, propynylene, butynylene, pentynylene, hexynylene, and the like.
  • Cycloalkyl refers to a saturated or partially unsaturated cyclic hydrocarbon.
  • the cycloalkyl ring may include the specified number of carbon atoms.
  • a 3- to 8-membered cycloalkyl group includes 3, 4, 5, 6, 7, or 8 carbon atoms.
  • suitable cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, 1,4-cyclohexadienyl, cycloheptyl and Cyclooctyl.
  • cycloalkylene refers to a divalent cyclic hydrocarbon group derived from a cycloalkyl group. E.g Wait.
  • alkoxy refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), where the definition of alkyl is as described above.
  • alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy.
  • the alkoxy group may be optionally substituted or unsubstituted.
  • the substituent is preferably one or more of the following groups, which are independently selected from H atom, D atom, halogen, alkyl group, alkoxy group , Haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl substituted by one or more substituents.
  • aryl refers to any stable, monocyclic or bicyclic carbocyclic ring of up to 7 atoms in each ring, at least one of which is aromatic. Examples of such aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, or binaphthyl.
  • heterocycloalkyl or “heterocyclyl” refers to a cyclic hydrocarbon in which one to four carbon atoms have been independently selected from N, N(R), S, S(O), S(O) 2 and O ⁇ heteroatom replacement.
  • the heterocyclic ring can be saturated or unsaturated.
  • heterocyclic groups examples include tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, pyrrolinyl, pyrazolinyl, pyranyl, piperidinyl, pyrazolinyl, dithiolanyl , Oxathiolanyl, dioxanyl, dioxanyl, morpholino and oxazinyl.
  • heteroaryl means a stable monocyclic or bicyclic ring of up to 7 atoms in each ring, wherein at least one ring is aromatic and at least one ring contains 1 to 4 heteroatoms selected from O, N, and S.
  • Heteroaryl groups within the scope of this definition include, but are not limited to, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, quinazolinyl, pyrazolyl, indolyl, benzotriazolyl, furan Group, thienyl, phenylthio, 3,4-propylenedioxythiophenyl, benzothienyl, benzofuranyl, benzodioxane, benzodioxa Cyclohexene, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, imidazolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, tetrahydroquinoline, thiazolyl, Isothiazolyl, 1,2,4-triazolyl, 1,2,3-triazolyl, 1,
  • halogen refers to fluorine, chlorine, bromine or iodine.
  • halo refers to substitution by one or more atoms selected from fluorine, chlorine, bromine, and iodine.
  • Dendrimer refers to a molecule containing a core and at least one dendrimer attached to the core.
  • Each dendron is composed of at least one layer or one generation of branched structural units, resulting in more and more branched structures with branches of each generation of structural units.
  • the maximum number of dendrons attached to the core is limited by the number of functional groups on the core.
  • Structural unit refers to a branched molecule with at least three functional groups. One functional group is used to connect to the core or the previous generation of the structural unit and at least two functional groups are used to connect to the next generation of the structural unit or form a dendrimer. surface.
  • “Generation” refers to the number of layers of structural units constituting the dendrimer or dendrimer.
  • a first-generation dendrimer will have a layer of branched structural units connected to the core, for example, core-[[structural unit]] u , where u is the number of dendrons connected to the core.
  • Two-generation dendrimers have two layers of structural units in each dendrimer connected to the core.
  • the dendritic polymer may be: core [[structural unit] [Structural unit] 2 ] u .
  • the third-generation dendrimer has a three-layer structure unit in each dendrimer connected to the core, for example, core-[[structural unit][structural unit] 2 [structural unit] 4 ]u.
  • Six-generation dendrimers have six layers of structural units connected to the core, such as core-[[structural unit][structural unit] 2 [structural unit] 4 [structural unit] 8 [structural unit] 16 [structural unit] 32 ] u etc.
  • the last generation (outermost generation) of the structural unit provides the surface functionalization of the dendrimer and the number of functional groups available for binding end groups.
  • the outermost generation has 64 structural units and 128 functional groups can be used to bind end groups.
  • “Slightly soluble” refers to a drug or pharmaceutically active agent that has a solubility of 1 mg/mL to 10 mg/mL in water. Drugs with a solubility of less than 1 mg/mL in water are considered insoluble.
  • Solubilizing excipient refers to a formulation additive used to dissolve an insoluble or slightly soluble pharmaceutically active agent in an aqueous formulation.
  • examples include surfactants such as polyethoxylated castor oil including Cremophor EL, Cremophor RH40 and Cremophor RH60, D- ⁇ tocopherol-polyethylene glycol 1000 succinate, polysorbate 20, polysorbate 80, solutol HS15 , Sorbitan monooleate, poloxamer 407, Labrasol, etc.
  • Optional or “optional” means that the event or environment described later can but need not occur, and the description includes occasions where the event or environment occurs or does not occur.
  • “L is a C 1-10 linear alkylene group optionally interrupted by one or more oxygen, sulfur or nitrogen atoms” means that the C 1-10 linear alkylene group may be interrupted by an oxygen, sulfur or nitrogen atom. However, it is not necessary to be interrupted. The description includes the case where the C 1-10 linear alkylene group is interrupted by an oxygen, sulfur or nitrogen atom and the case where the C 1-10 linear alkylene group is not interrupted by an oxygen, sulfur or nitrogen atom.
  • Substituted refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently of each other, substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine possible or impossible substitutions (by experiment or theory) without too much effort.
  • the bond Does not specify the configuration, that is, the key Can be Or both Two configurations.
  • the bond If the configuration is not specified, it can be the Z configuration or the E configuration, or both configurations can be included.
  • Atoms that can be isotopically labeled include, but are not limited to, hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, iodine, and the like. They can be replaced by isotopes 2 H(D), 3 H, 11 C, 13 C, 14 C, 15 N, 18 F, 31 P, 32 P, 35 S, 36 Cl and 125 I, respectively. Unless otherwise specified, when a position is specifically designated as deuterium (D), the position should be understood as having an abundance of deuterium (ie, at least 45 % Deuterium incorporation).
  • the pharmaceutically active agent or its residue, pharmaceutically active agent, pharmaceutically active agent or its residue A can be used interchangeably, and all refer to molecules or groups with pharmacological activity.
  • Figure 1 is a curve of the human plasma conversion concentration of the compound of the present disclosure in Test Example 1.
  • Figure 2 is a beagle plasma PK curve in Test Example 2.
  • the experimental methods that do not specify specific conditions in the embodiments of the present invention usually follow conventional conditions or the conditions recommended by raw material or commodity manufacturers.
  • the reagents without specific sources are the conventional reagents purchased on the market.
  • the structure of the compound is determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR is measured with Bruker AVANCE-400 nuclear magnetic instrument, the solvent is deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl 3 ), deuterated methanol (CD 3 OD), and the internal standard is tetramethyl sulfoxide (DMSO-d6) Based on silane (TMS), the chemical shift is given in units of 10 -6 (ppm).
  • NMR nuclear magnetic resonance
  • MS mass spectrometry
  • HPLC high performance liquid chromatography
  • the thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the size of the silica gel plate used for thin layer chromatography (TLC) is 0.15mm ⁇ 0.2mm, and the size of the thin layer chromatography separation and purification product is 0.4mm. ⁇ 0.5mm silica gel plate.
  • the known starting materials of the present invention can be synthesized by or according to methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organnics, Aldrich Chemical Company, Accela ChemBio Inc, Darui Chemicals and other companies.
  • the reactions are all carried out under an argon atmosphere or a nitrogen atmosphere.
  • the argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1L.
  • the hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1L.
  • the pressure hydrogenation reaction uses Parr 3916EKX hydrogenator and Qinglan QL-500 hydrogen generator or HC2-SS hydrogenator.
  • the hydrogenation reaction is usually evacuated and filled with hydrogen, and the operation is repeated 3 times.
  • the microwave reaction uses the CEM Discover-S 908860 microwave reactor.
  • the solution in the reaction refers to an aqueous solution.
  • reaction temperature is room temperature.
  • Room temperature is the most suitable reaction temperature, and the temperature range is 20°C to 30°C.
  • the preparation of the PBS buffer with pH 6.5 in the examples: take KH 2 PO 4 8.5g, K 2 HPO 4 .3H 2 O 8.56g, NaCl 5.85g, and EDTA 1.5g in a bottle, dilute the volume to 2L, and ultrasonic Dissolve all of it, shake well and get it.
  • the eluent system of column chromatography and the developing solvent system of thin-layer chromatography used to purify compounds include: A: dichloromethane and isopropanol system, B: dichloromethane and methanol system, C: petroleum ether and In the ethyl acetate system, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of triethylamine and acidic or alkaline reagents can also be added for adjustment.
  • Q-TOF LC/MS uses Agilent 6530 accurate mass quadrupole-time-of-flight mass spectrometer and Agilent 1290-Infinity ultra-high performance liquid chromatograph (Agilent Poroshell 300SB-C8 5 ⁇ m, 2.1 ⁇ 75mm column).
  • the dendrimer shown in the following examples includes the core of the reference dendrimer and the structural units in the outermost generation of the dendrimer.
  • the first generation to the subsurface generation are not depicted.
  • Dendrimer BHALys[Lys] 32 represents the fifth generation dendrimer, which has the following formula BHALys[Lys] 2 [Lys] 4 [Lys] 8 [Lys] 16 [Lys] 32 , 64 surface amino groups can be used for bonding ⁇ On the end group.
  • 1-A00, 1-B00, and 1-C00 are synthesized according to patent WO2012167309A, and the pharmaceutically active agent attached is docetaxel; 2-A00 is synthesized according to patent WO2018154004A, and the attached pharmaceutically active agent is compound 2.
  • Dendrimer stent BHALys[Lys] 32 [ ⁇ -NH 2 ⁇ TFA] 32 [ ⁇ -PEG 1100 ] 32 is referred to as Dendrimer 1
  • BHALys[Lys] 32 [ ⁇ -NH 2 ⁇ TFA] 32 [ ⁇ -PEG 2100 ] 32 is referred to as Dendrimer 1-PEG2K in the examples, synthesized according to the method of WO2018154004A:
  • the macromolecules in the present disclosure can be synthesized according to a route selected from the following, and PG is a carboxylic acid protecting group:
  • compound 1-Q01 (60.6 mg, 63.9 ⁇ mol) and PyBOP (63.6 mg, 122.3 ⁇ mol) were dissolved in anhydrous DMF (1.5 mL), and compound Dendrimer 1 (72 mg, 1.59 ⁇ mol) and DIPEA ( 15.8 mg, 102.3 ⁇ mol) of anhydrous DMF (1.5 mL) mixed solution, reacted for 1.5 hours.
  • the reaction solution was directly purified by column chromatography to obtain 80 mg of oily compound. It was dissolved in purified water (20 mL), filtered through a filter membrane (0.22 ⁇ m), and lyophilized to obtain 78 mg of product 1-Q00 with a yield of 76%.
  • Tetrahydrofuran (300 mL) was added to the 1-T01-6B aqueous solution (111.8 g, 947 mmol) obtained in the previous step, cooled to 0° C., tert-butyl bromoacetate (92.4 g, 663 mmol) was added dropwise, and then heated to room temperature and stirred for 20 hours.
  • the tetrahydrofuran was removed under reduced pressure, the pH was adjusted to 3-4 with 2N hydrochloric acid, extracted with ethyl acetate, concentrated and purified by column chromatography to obtain 49.3 g of crude product.
  • 250 ml of petroleum ether was added, stirred at room temperature for 2 hours, filtered and dried to obtain 25.0 g of product 1-T01-tBu, with a total yield of 11.1% in the four steps.
  • the compounds docetaxel (80mg, 0.1mmol) and 1-T01-P (53mg, 0.2mmol) were dissolved in dichloromethane (3mL), and DMAP (14mg, 0.12mmol) and DCC (25mg , 0.12mmol), stirred at room temperature overnight. After concentration, it was purified by column chromatography to obtain 70 mg of product 1-T01-Bn with a yield of 67%.
  • compound 1-Z01-A-6 (220 mg, 1.3 mmol, synthesized according to the literature Molecules, 2020, 25, 2153) was dissolved in tetrahydrofuran (5 mL), cooled to 0° C., and NaH (104 mg, 2.2mmol, 60% in oil), control the internal temperature ⁇ 20°C, and stir for 15 minutes.
  • a tetrahydrofuran solution of compound S2 (360 mg, 1.6 mmol) was added dropwise, and the mixture was stirred at room temperature overnight.
  • reaction solution was cooled to 0°C, quenched with saturated aqueous NaHCO 3 (10 mL), extracted with ethyl acetate (10 mL ⁇ 2), concentrated and purified by column chromatography to obtain 180 mg of product 1-Z01-A-7. The yield was 43%.
  • compound 1-Z01-A-7 (180 mg, 0.56 mmol) was dissolved in dichloromethane (2 mL), cooled to 0° C., and trifluoroacetic acid (2 mL) was added. Warm to room temperature and stir overnight. After concentration, it was purified by column chromatography to obtain 50 mg of product 1-Z01-A-P with a yield of 32%.
  • the compounds docetaxel (120mg, 0.15mmol) and 1-Z01-AP (48mg, 0.18mmol) were dissolved in DMF (1mL), and DMAP (19mg, 0.15mmol) and DCC (38mg, 0.18 mmol), stirring at room temperature overnight. After concentration, it was purified by column chromatography to obtain 50 mg of product 1-Z01-A-Bn with a yield of 32%.
  • compound 1-Z01-B-4 (10.3g, 47.9mmol, synthesized according to patent WO2016205633A) was dissolved in DMF (150mL), potassium carbonate (33.1g, 239.5mmol) and benzyl bromide (10.6g) were added , 62.3mmol), heated to 60°C, and reacted for 16 hours. After cooling, it was quenched with water (300 mL), extracted with ethyl acetate, concentrated and purified by column chromatography to obtain 13 g of compound 1-Z01-B-5 with a yield of 89%.
  • compound 1-Z01-B (573 mg, 594.8 ⁇ mol) and PyBOP (596 mg, 1145 ⁇ mol) were dissolved in anhydrous DMF (9.8 mL), and compound Dendrimer 1 (700 mg, 14.87 ⁇ mol) and DIPEA (148 mg) were added dropwise. , 1145 ⁇ mol) of anhydrous DMF (9.8mL) mixed solution, react for 2 hours.
  • reaction solution was diluted with acetonitrile (19.6 mL), it was passed through ultrafiltration (10KD, ) Device, purified by acetonitrile ultrafiltration to obtain 1.01 g crude product, dissolved in purified water (100 mL), filtered through a filter membrane (0.22 ⁇ m), and freeze-dried to obtain 0.99 g of product 1-Z00-B with a yield of 100%.
  • ultrafiltration (10KD, ) Device purified by acetonitrile ultrafiltration to obtain 1.01 g crude product, dissolved in purified water (100 mL), filtered through a filter membrane (0.22 ⁇ m), and freeze-dried to obtain 0.99 g of product 1-Z00-B with a yield of 100%.
  • compound 1-Z01-B4-4 (6.7g, 36.0mmol, synthesized according to literature Molecules, 2020, 25, 2153) and anhydrous potassium carbonate (10.0g, 72.4mmol) were added to anhydrous acetonitrile (67mL ), add benzyl bromoacetate (9.9 g, 43.2 mmol), and react at room temperature for 16 hours. After filtration and concentration, the crude product was purified by column chromatography to obtain 10.1 g of 1-Z01-B4-5-Bn with a yield of 88%.
  • compound 2 (4.25g, 4.5mmol, synthesized according to patent WO2012017251) was dissolved in dichloromethane (70mL), DMAP (165mg, 1.35mmol) and compound 1-Z01-B4 (1.85g, 8.1mmol) ), cooled to 0°C, added dropwise a solution of DCC (1.67g, 8.1mmol) in DCM (10mL), warmed to room temperature and stirred overnight. After filtration, it was concentrated under reduced pressure, and the crude product was separated by column chromatography to obtain 4.2 g of product 2-B00-1 with a yield of 80%.
  • TMSOTf Trimethylsilyl fluoromethanesulfonate
  • compound 2-B00-2 (1.38g, 1.25mmol) and PyBOP (0.79g, 1.51mmol) were dissolved in anhydrous DMF (21mL), and compound Dendrimer 1-PEG2K (2.00g, 0.025mmol) was added dropwise.
  • compound Dendrimer 1-PEG2K (2.00g, 0.025mmol) was added dropwise.
  • NMM N-methylmorpholine
  • reaction solution was diluted with acetonitrile (42mL), it was passed through ultrafiltration (10KD, ) Device, purified by acetonitrile ultrafiltration to obtain 2.25 g crude product, dissolved in purified water (150 mL), filtered through a filter membrane (0.22 ⁇ m), and lyophilized to obtain 2.20 g product 2-B00 with a yield of 89%.
  • ultrafiltration (10KD, ) Device purified by acetonitrile ultrafiltration to obtain 2.25 g crude product, dissolved in purified water (150 mL), filtered through a filter membrane (0.22 ⁇ m), and lyophilized to obtain 2.20 g product 2-B00 with a yield of 89%.
  • Test Example 1 Study on the stability of the compound of the present disclosure in human and mouse plasma
  • test compound and docetaxel solution were prepared in PBS.
  • a 1 mM propane theophylline working solution was prepared in acetonitrile.
  • propane theophylline was used as a positive control for human and mouse plasma.
  • the sample is analyzed by LC-MS/MS
  • peak area ratio t hour is the peak area ratio of the test compound at t;
  • the peak area ratio STD is the peak area ratio of 200 ⁇ g/mL docetaxel.
  • ⁇ c refers to the difference in plasma conversion concentration between the last two time points
  • the compound of the present disclosure has the effect of smoothly releasing the drug. Among them, the burst release of compound 1-S00 is lower, and the drug release is better. On the basis of reaching the therapeutic concentration, the release is more stable within 24 hours (can be compared from the ⁇ c value in the table), modified by sustained drug release The in vivo pharmacokinetic characteristics can reduce side effects and increase the therapeutic window.
  • Test Example 2 Pharmacokinetic study of different compounds in beagle dogs administered by a single intravenous injection
  • Sample preparation method 120 mg/bottle of test substance, add final volume of 5% DMSO, 95% normal saline, and vortex to dissolve to obtain 1.5 mg/ml test sample.
  • the collected blood samples are placed in EDTA-K2 anticoagulant blood collection tubes. All blood samples need to be added with esterase inhibitors for special treatment.

Abstract

涉及一类载药的大分子及其制备方法。具体地，该大分子是负载药物及药代动力学修饰剂的树枝状聚合物，尤其涉及通过特定连接子将药物连接于树枝状聚合物。该类大分子可以用于对药物的释放速率进行调节，具体是通过连接子的选择进行的。

Description

一类载药的大分子及其制备方法 技术领域
本公开属于医药领域，涉及负载药物及药代动力学修饰剂的树枝状聚合物，尤其是涉及通过特定连接子将药物连接于树枝状聚合物。
背景技术
目前，虽然药物的研究开发取得了长足进步，但还是有许多药物在临床试验阶段因其物理性质(比如溶解性)使得其难以配制成合适的制剂进行给药，或由于给药后发生的高药物浓度期间导致的毒性效应及差的治疗指数而失败。此外，还包括其它缺点如吸收差、生物利用度低、体内稳定性差、靶向性差导致的全身性副作用，以及在给药后无法控制其在体内的生物分布、代谢和肾或肝清除。随着药物研究的不断进步，出现了一些新的研究领域和一些具有重大潜力的技术方法来推动药物的开发，诸如在脂质体、胶束或聚合胶束制剂中配制药物试剂，以及将药物试剂共价附着至亲水聚合物主链等。虽然这些策略可增溶药物活性剂且在某些情况下改善生物利用度和靶向性，但药物活性剂的释放存在困难，在某些情况下，在药物分子到达靶向器官之前，载体就会快速降解释放出药物活性剂。很多情况下药物活性剂从载体的释放速率为可变的，因此使药物在体内或在靶器官中达不到有效治疗剂量。
近年来，已发现树枝状聚合物在生物技术和药物应用领域取得明显进步(Xiangyang Shi et al.Sci China Mater,2018,61(11),1387–1403.)。树枝状聚合物是具有密集分支结构的特定种类的聚合物，从核心分子出发不断地向外重复支化生长而得到的结构类似于树状的大分子，即核心经过分支长到一定长度后以分成两个分枝，如此重复进行，直到长得足够稠密以致于长成像球形一样的树丛(V Gajbhiye et al.Journal of Pharmacy and Pharmacology,2009,61,989–1003.)，其特征在于比普通聚合物更高浓度的官能团/单位分子体积。特别是树枝状聚合物的独特性质，例如其高分支程度、多价、球形结构和良好确定的分子量，使得其给予用于药物递送的新支架的希望。在过去十年中，关于生物相容性树枝状聚合物的设计和合成及其对生物科学的许多领域包括药物递送的应用的研究也越来越受关注。
澳大利亚的Starpahrma公司利用其开发的树枝状聚赖氨酸进行抗癌药物的负载和运输，来增强药物的药理性质，确保了药物在适当的时候被送到身体的适当部位，这种方法被称为“药物输送”，这项技术以
Figure PCTCN2021089174-appb-000001
商标面市。其中
Figure PCTCN2021089174-appb-000002
Figure PCTCN2021089174-appb-000003
以及
Figure PCTCN2021089174-appb-000004
是该公司目前利用
Figure PCTCN2021089174-appb-000005
技术重点开发处于临床研究阶段的3款抗癌药物，显示了很好的开发前景。
CN103796684A公开了通过二酸连接子将药物与树枝状聚合物连接起来的大分子，尤其是通过包含被氧、氮或硫原子间断的C 1-C 10的饱和支链或直链的二酸 连接子连接。
发明内容
本公开提供了一种大分子，其包含：
i)具有表面氨基的树枝状聚合物D，其中至少两个不同的端基共价连接于所述树枝状聚合物的表面氨基：
ii)第一端基，其为包含羟基、氨基、羧基或者巯基的药学活性剂或其残基A；
iii)第二端基，其为药代动力学修饰剂；
其中所述第一端基通过连接子-X 1-L-X 2-共价连接于所述树枝状聚合物的表面氨基，X 1为连接子与药学活性剂的连接基，X 2为连接子与树枝状聚合物的连接基，其中：
a)X 1和X 2选自-C(O)-或不存在，L为任选被一个或多个氧、硫或氮原子间断的C 1-10直链或支链亚烷基；当X 1和X 2中至少一个不存在时，所述C 1-10直链或支链亚烷基任选被取代基取代；当X 1和X 2均为-C(O)-时，所述C 1-10直链或支链亚烷基进一步被取代基取代；所述的取代基选自氘、羟基、C 3-7环烷基、C 3-7亚环烷基、烷基、烷氧基、卤代烷基、卤代烷氧基、卤素、硝基、氰基、酰基、硫醚基、亚磺酰基、磺酰基、-NR 1R 2、芳基、杂芳基和杂环基的一个或多个基团；或者
b)X 1和X 2选自-C(O)-或不存在，L为任选被一个或多个氧、硫或氮原子间断的C 1-10直链或支链亚烯基或亚炔基，所述亚烯基和亚炔基任选被选自氘、羟基、烷基、环烷基、烷氧基、卤代烷基、卤代烷氧基、卤素、硝基、氰基、酰基、硫醚基、亚磺酰基、磺酰基、-NR 1R 2、芳基、杂芳基和杂环基的一个或多个基团所取代，或者
c)X 1选自-C(O)-或不存在，X 2不存在，L与树枝状聚合物表面氨基的氮原子连接后形成含氮杂环结构
Figure PCTCN2021089174-appb-000006
L 1为任选被一个或多个氧、硫或氮原子间断的C 1-10直链或支链亚烷基、亚烯基或亚炔基，所述亚烷基、亚烯基或亚炔基任选被选自氘、羟基、烷基、环烷基、亚环烷基、烷氧基、卤代烷基、卤代烷氧基、卤素、硝基、氰基、酰基、硫醚基、亚磺酰基、磺酰基、-NR 1R 2、芳基、杂芳基和杂环基的一个或多个基团所取代，X 3选自CR 5或N，其中R 5选自氢、氘、羟基、烷基或烷氧基，m选自0、1或2；或者
d)X 1选自-C(O)-或不存在，X 2不存在，且L为
Figure PCTCN2021089174-appb-000007
L 2为任选被一个或多个氧、硫或氮原子间断的C 1-10直链或支链亚烷基、亚烯基或亚炔基，所述 亚烷基、亚烯基或亚炔基任选被选自氘、羟基、烷基、环烷基、烷氧基、卤代烷基、卤代烷氧基、卤素、硝基、氰基、酰基、硫醚基、亚磺酰基、磺酰基、-NR 1R 2、芳基、杂芳基和杂环基的一个或多个基团所取代，n选自1或2，其中含氮杂环端与树枝状聚合物连接，L 2端通过-C(O)-或者直接与药学活性剂连接；
其中，R 1、R 2各自独立地选自氢、羟基、C 1-6烷基、环烷基、C 1-6烷氧基。
在一些实施方案中，X 1和X 2均为-C(O)-，L为被一个或多个氧、硫或氮原子间断的C 1-6直链或支链亚烷基，所述C 1-6直链或支链亚烷基被独立地选自氘、羟基、C 3-7环烷基、C 3-7亚环烷基、C 1-6烷氧基、C 1-6卤代烷基、C 1-6卤代烷氧基和卤素的一个或多个取代基所取代。
在一些实施方案中，X 1和X 2均为-C(O)-，L为被一至三个独立地选自硫或氮的原子间断的C 1-6直链或支链亚烷基，所述C 1-6直链或支链亚烷基被独立地选自羟基、C 3-7环烷基、C 3-7亚环烷基、C 1-6烷氧基、C 1-6卤代烷基、C 1-6卤代烷氧基和卤素的一个或多个取代基所取代。
在一些实施方案中，所述C 1-6直链或支链亚烷基被一个或多个C 3-7亚环烷基取代。
在一些实施方案中，连接子为-C(O)-X-C(O)-，
其中X为
Figure PCTCN2021089174-appb-000008
R a、R b各自独立地选自氢、C 1-6烷基，或R a、R b与其相连的碳原子一起形成C 3-7环烷基；R c、R d各自独立地选自氢、C 1-6烷基，或R c、R d与其相连的碳原子一起形成C 3-7环烷基；条件是至少有一个R a、R b与其相连的碳原子一起形成C 3-7环烷基或至少有一个R c、R d与其相连的碳原子一起形成C 3-7环烷基；
X’选自-CH 2-、-O-、-S-和-NR 3-；
R 3为氢或C 1-6烷基；
s和t各自独立地选自1至4的整数；
R a、R b、R c、R d在每次出现时可选择相同或不同的基团。
在一些实施方案中，X为
Figure PCTCN2021089174-appb-000009
Figure PCTCN2021089174-appb-000010
其中s1和t1各自独立地选自1至4的整数。
在一些实施方案中，X’选自-O-、-S-、-NH-和-N(CH 3)-。
在一些实施方案中，连接子-X 1-L-X 2-中，X 1为-C(O)-，与药学活性剂或其残基A连接；X 2为-C(O)-，与树枝状聚合物D的表面氨基连接形成酰胺键。
在一些实施方案中，药学活性剂或其残基A包含羟基，并与X1形成酯键。
在一些具体实施方案中，连接子-X 1-L-X 2-中，X 1为-C(O)-，与药学活性剂或其残基A连接；X 2为-C(O)-，与树枝状聚合物D的表面氨基连接形成酰胺键，结构选自如下：
Figure PCTCN2021089174-appb-000011
在一些实施方案中，连接子-X 1-L-X 2-中，X 1为-C(O)-，与药学活性剂或其残基A连接；X 2为-C(O)-，与树枝状聚合物D的表面氨基连接形成酰胺键，结构如下：
Figure PCTCN2021089174-appb-000012
在一些实施方案中，连接子-X 1-L-X 2-中，X 1为-C(O)-，与药学活性剂或其残基A连接；X 2不存在，L的一部分基团形成氮杂环，与树枝状聚合物D的表面氨基连接形成碳-氮键。
在一些具体实施方案中，连接子-X 1-L-X 2-中，X 1为-C(O)-，与药学活性剂或其残基A连接；X 2不存在，L的一部分基团形成氮杂环，与树枝状聚合物D的表面氨基连接形成碳-氮键，结构选自如下：
Figure PCTCN2021089174-appb-000013
本公开还提供一种大分子，其包含：
i)具有表面氨基的树枝状聚合物，其中至少两个不同的端基共价连接于所述树枝状聚合物的表面氨基：
ii)第一端基，其为包含羟基、氨基、羧基或者巯基的药学活性剂或其残基；
iii)第二端基，其为药代动力学修饰剂；
其中所述第一端基通过三酸连接子共价连接于所述树枝状聚合物的表面氨基， 三酸连接子中的一个羰基与药学活性剂或其残基连接，另外两个羰基与树枝状聚合物的同一表面氨基连接并因此与连接子部分形成含氮杂环。
在一些实施方案中，所述的三酸连接子与药学活性剂或其残基A以及树枝状聚合物D连接，形成如下的结构：
Figure PCTCN2021089174-appb-000014
其中X 4选自C、N，L 3为任选被氧、硫或氮原子间断的C 1-10亚烷基、亚烯基或者亚炔基，p为选自0-5的整数。
在一些实施方案中，所述的三酸连接子与药学活性剂或其残基A以及树枝状聚合物D连接，形成选自如下的结构：
Figure PCTCN2021089174-appb-000015
其中，D为树枝状聚合物，A为药学活性剂或其残基。
在一些实施方案中，所述的药学活性剂或其残基含有羟基，并与三酸连接子的羰基形成酯键。
本公开还提供一种大分子，其包含：
i)具有表面氨基的树枝状聚合物D，其中至少两个不同的端基共价连接于所述树枝状聚合物的表面氨基：
ii)第一端基，其为艾日布林、曲贝替定或卢比替定(Lurbinectedin)；
iii)第二端基，其为药代动力学修饰剂；
其中所述第一端基通过二酸连接子共价连接于所述树枝状聚合物的表面氨基，所述二酸连接子包含任选被一个或多个硫或氮原子间断的C 1-10烷基链。
在一些实施方案中，所述的二酸连接子包含的C 1-10烷基链未被氧、硫或氮原子间断。
在特定的实施方案中，二酸连接子可选自
Figure PCTCN2021089174-appb-000016
Figure PCTCN2021089174-appb-000017
在一些实施方案中，所述的二酸连接子包含被一个或多个氧、硫或氮原子间断的C 1-10烷基。
在特定的实施方案中，二酸连接子可选自
Figure PCTCN2021089174-appb-000018
Figure PCTCN2021089174-appb-000019
本公开还提供一种大分子，其包含：
i)具有表面氨基的树枝状聚合物D，其中至少两个不同的端基共价连接于所述树枝状聚合物的表面氨基：
ii)第一端基，其为包含羟基、氨基、羧基或者巯基的药学活性剂或其残基，并且不是多西他赛；
iii)第二端基，其为药代动力学修饰剂；
其中所述第一端基通过选自如下的连接子共价连接于所述树枝状聚合物的表面氨基：
Figure PCTCN2021089174-appb-000020
在一些实施方案中，选择本公开的连接子可以提供所需的药物释放速率，例如，快速释放或者缓慢释放。
在一些实施方案中，大分子的药学活性剂的释放速率比不依赖于大分子递送的快一些，可能至少快1倍。在一些实施方案中，大分子的药学活性剂的释放速率比不依赖于大分子递送的释放的更慢，可能慢两倍、三倍、四倍、五倍、六倍、七倍、八倍、九倍、十倍或者十倍以上、十五倍以上、二十倍以上、三十倍以上。低释放速率的大分子适合将大分子配制成长时间例如1周到3个月、1个月到6个月、6个月以上的时间内缓慢释放的药物。快速释放优选在0-8h内，尤其是0-4h内，特别是0-2h，更特别是5-60分钟内释放大于50％的药学活性剂。中等释放优选在1-72小时内，尤其是2-48小时内释放大于50％的药学活性剂。药学活性剂的释放速率可以通过选择合适的连接子进行控制，释放速率还依赖药物活性剂的特性。在一些实施方案中，药学活性剂通过相同的连接子连接于树枝状聚合物。在另一些实施方案中，药学活性剂通过两种或者多种连接子与树枝状聚合物进行连接，以使药学活性剂可以以不同的释放速率从大分子释放。
在一些实施方案中，所述的第一端基与第二端基以1:2-2:1的比率存在，尤其是1:2、1:1、2:1。在一些实施方案中，所述大分子包含为封闭基团、药物或者靶向基团的第三端基。封闭基团可以是酰基。在一些实施方案中，第一端基、第二端基和第三端基的比率为1:1:1-1:2:2，尤其是1:2:1。在一些实施方案中，其中至少50％的端基包含第一端基或者第二端基中的一者。在特定实施方案中，药物活性剂结合至大于14％、25％、27％、30％、39％、44％或48％的表面氨基。在一些实施方案中，药代动力学修饰剂结合至大于15％、25％、30％、33％或46％的表面氨基的总数。
本公开所述药学活性剂可选自：麻醉剂、抗酸剂、抗体、抗感染药、生物制品、心血管药物、造影剂、利尿剂、补血药、免疫抑制剂、激素和类似物、营养制品、眼科药物、疼痛治疗剂、呼吸药物、佐剂、同化剂、抗关节炎药、抗惊厥药、抗组胺药、消炎药、抗溃疡药、行为矫正药物、肿瘤药物、中枢神经系统药物、避孕药、糖尿病治疗药物、生育药物、生长促进剂、止血药、免疫刺激剂、肌肉松弛药、肥胖治疗剂、骨质疏松症药物、肽、镇静剂和安定药、尿道酸化剂或维生素。
在一些实施方案中，药物活性剂为肿瘤药物、类固醇、阿片类镇痛药、呼吸系统药物、中枢神经系统(CNS)药物、高胆固醇血症药物、抗高血压药物、抗生素、免疫抑制药物、抗菌剂、促黄体激素释放激素(LHRH)激动剂、LHRH拮抗剂、抗病毒药、抗逆转录病毒药物、雌激素受体调节剂、生长抑素类似物、抗炎药物、维生素D2类似物、合成甲状腺素、抗组胺剂、抗真菌剂或非类固醇类抗炎药物(NSAID)。
在一些实施方案中，所述的肿瘤药物包括紫杉烷(如紫杉醇、卡巴他赛和多 西他赛)、喜树碱及其类似物(如伊立替康和托泊替康)、其它抗微管剂(如长春氟宁)、核苷类(如吉西他滨、克拉屈滨、氟达拉滨、卡培他滨、地西他滨、阿扎胞苷、氯法拉滨和奈拉滨)、激酶抑制剂(如达沙替尼(sprycel)、坦罗莫斯(temisirolimus)、达沙替尼、AZD6244、AZD1152、PI-103、R-roscovitine、奥罗莫星和purvalanol A)和埃博霉素B类似物(如伊沙匹隆)、蒽环类药物(anthrocyclines)(如氨柔比星、多柔比星、表柔比星和戊柔比星)、海鞘素衍生物(如曲贝替定(trabectecin)、卢比替定(lurbinectedin))、蛋白酶体抑制剂(如硼替佐米)和其它拓扑异构酶抑制剂、嵌入剂和烷化剂、微管抑制剂如艾日布林等，或这些药物分子的结构改造物。
在一些实施方案中，所述的药学活性剂选自紫杉烷类药物、喜树碱衍生物、核苷类药物、蒽环类药物、海鞘素衍生物、蛋白酶体抑制剂、微管抑制剂、BCL-2抑制剂、BCL-X L抑制剂、选择性核输出抑制剂、抗代谢药物、酪氨酸激酶抑制剂、PLK1抑制剂、CDK4/6抑制剂、BTK抑制剂、非甾体激素受体拮抗剂和类固醇，优选紫杉烷类药物、BCL-2抑制剂和BCL-X L抑制剂。
在一些实施方案中，所述的类固醇包括合成类固醇(如睾酮、双氢睾酮和乙炔雌二醇)和皮质类固醇(如可的松、泼尼松龙(prednisilone)、布地奈德、曲安西龙、氟替卡松、莫米松、安西奈德、氟轻松(flucinolone)、醋酸氟轻松(fluocinanide)、地奈德、氯氟舒松、泼尼卡酯、氟可龙、地塞米松、倍他米松和氟泼尼定(fluprednidine))。
在一些实施方案中，所述的阿片类镇痛药包括吗啡、羟吗啡酮、纳洛酮、可待因、羟考酮、甲基纳曲酮、氢吗啡酮、丁丙诺啡和埃托啡。
在一些实施方案中，所述的呼吸系统药物包括支气管扩张剂、吸入类固醇和减充血剂特别是沙丁胺醇、异丙托溴铵、孟鲁司特和福莫特罗。
在一些实施方案中，所述的CNS药物包括抗精神病药(如喹硫平)和抗抑郁药(如文拉法辛)。
在一些实施方案中，所述的高胆固醇血症药物包括依泽替米贝和他汀类如辛伐他汀、洛伐他汀、阿托伐他汀、氟伐他汀、匹伐他汀、普伐他汀和瑞舒伐他汀。
在一些实施方案中，所述的抗高血压药物包括氯沙坦、奥美沙坦、坦酯(medoxomil)、美托洛尔(metrolol)、曲伏前列素和波生坦。
在一些实施方案中，所述的免疫抑制药物包括糖皮质激素、细胞抑制剂、抗体片段、抗嗜免疫剂、干扰素、TNF结合蛋白特别是钙调磷酸酶抑制剂(cacineurin inhibitors)如他克莫司、霉酚酸及其衍生物，如吗替麦考酚酯和环孢菌素。
在一些实施方案中，所述的抗菌剂包括抗生素，如阿莫西林、美罗培南和克拉维酸。
在一些实施方案中，所述的LHRH激动剂包括醋酸戈舍瑞林、地洛瑞林和亮丙瑞林。在一些实施方案中，所述的LHRH拮抗剂包括西曲瑞克、加尼瑞克、阿巴瑞克和地加瑞克(degarelix)。
在一些实施方案中，所述的抗病毒药包括核苷类似物如拉米夫定、齐多夫定、阿巴卡韦和恩替卡韦，以及抗逆转录病毒药物包括蛋白酶抑制剂如阿扎那韦、lapinavir和利托那韦(ritonavir)。
在一些实施方案中，所述的雌激素受体调节剂包括雷洛昔芬和氟维司群。
在一些实施方案中，所述的生长抑素类似物包括奥曲肽。
在一些实施方案中，所述的抗炎药物包括美沙拉秦以及合适的NSAID包括扑热息痛(对乙酰氨基酚)。
在一些实施方案中，所述的维生素D2类似物包括帕立骨化醇。
在一些实施方案中，所述的合成甲状腺素包括左甲状腺素。
在一些实施方案中，所述的抗组胺剂包括非索非那定。
在一些实施方案中，所述的抗真菌剂包括唑类诸如伏立康唑(viriconazole)。
在一些实施方案中，所述的药学活性剂是艾日布林。在一些实施方案中，所述的药学活性剂是多西他赛。在一些实施方案中，所述的药学活性剂是曲贝替定。在一些实施方案中，所述的药学活性剂是卢比替定(lurbinectedin)。
在一些实施方案中，所述的药学活性剂选自多西他赛、伊立替康、吉西他滨、卡培他滨、地西他滨、阿扎胞苷、多柔比星、表柔比星、曲贝替定、卢比替定、硼替佐米、艾日布林、塞利尼索、维奈托克、特西他赛、培美曲塞、卡巴他赛、卡博替尼、昂凡色替(Onvansertib)、下式化合物2，或这些药物分子的结构改造物，
Figure PCTCN2021089174-appb-000021
在一些实施方案中，所述的药学活性剂在水溶液中是微溶或者不溶的。
第二端基为药代动力学修饰剂，可以修饰或调解药学活性剂或大分子的药代动力学特征，包括吸收、分布、代谢和/或排泄。在特定的实施方案中，药代动力学修饰剂延长药学活性剂的血浆半衰期，使得连接于大分子的药学活性剂的半衰期比单纯的药学活性剂或在非树枝状聚合物载体上的药学活性剂的半衰期更长。优选地，大分子或组合物的半衰期比单纯的药学活性剂或在非树枝状聚合物载体 上的药学活性剂的半衰期长至少2倍，且更优选地至少10倍。
所述的药代动力学修饰剂可选自聚乙二醇、聚乙基噁唑啉、聚乙烯基吡咯烷酮、聚丙二醇、叶酸盐或者关于细胞表面受体的配体的叶酸盐衍生物。在一些实施方案中，所述的药代动力学修饰剂是聚乙二醇。在一些实施方案中，所述的聚乙二醇具有220至5500Da范围内的分子量，例如可以是220-2500Da、570-2500Da、220-1100Da、570-1100Da、1000-5500Da、1000-2500Da、1000-2300Da的分子量。在一些实施方案中，所述的药代动力学修饰剂与所述树枝状聚合物表面氨基形成酰胺键。
靶向基团为结合至具有一些选择性的生物靶细胞、器官或组织的试剂，从而有助于将大分子导向体内特定的靶标并使其在该靶细胞、器官或组织中积聚。另外，靶向基团可以为通过受体介导的胞吞作用积极进入细胞或组织的大分子提供机制。特定的实例包括凝集素和抗体以及其它针对细胞表面受体的配体(包括小分子)。该相互作用可通过任何类型的键或缔合(包括共价键、离子键和氢键、范德华力)进行。合适的靶向基团包括结合于细胞表面受体的那些，例如，叶酸受体、肾上腺素能受体、生长激素、促黄体激素受体、雌激素受体、表皮生长因子受体、成纤维细胞生长因子受体(如FGFR2)、IL-2受体、CFTR和血管上皮生长因子(VEGF)受体。
在一些实施方案中，靶向基团为促黄体激素释放激素(LHRH)或其结合于促黄体激素释放激素受体的衍生物。在一些实施方案中，靶向基团为LYP-1，其为靶向肿瘤的淋巴系统而非正常组织的淋巴系统的肽。在一些实施方案中，靶向基团可为RGD肽。RGD肽为含有序列-Arg-Gly-Asp-的肽，该序列为细胞外基质蛋白质中主要的整联蛋白识别位点。在一些实施方案中，靶向基团可为叶酸。雌激素也可用于表达雌激素受体的靶细胞。
在一些实施方案中，靶向基团可直接地或优选地通过连接基团结合至树枝状聚合物核心。连接基团可为能够结合至核心的官能团和靶向基团上的官能团的任何二价基团。
本公开的大分子包含树枝状聚合物，其中结构单元的最外代具有表面氨基。大分子的树枝状聚合物的特性并不特别地重要，条件是它具有表面氨基。例如，树枝状聚合物可以为聚赖氨酸、聚赖氨酸类似物、聚酰氨基胺(PAMAM)、聚乙烯亚胺(PEI)或聚醚羟胺(PEHAM)树枝状聚合物。在一些实施方案中，树枝状聚合物是聚赖氨酸或聚赖氨酸类似物。聚赖氨酸或聚赖氨酸类似物包含核心和2-7代赖氨酸或赖氨酸类似物，例如包含2代、3代、4代、5代、6代或者7代赖氨酸或赖氨酸类似物。
在一些实施方案中，所述的赖氨酸具有如1所示结构：
Figure PCTCN2021089174-appb-000022
在一些实施方案中，所述赖氨酸类似物具有如2所示结构：
Figure PCTCN2021089174-appb-000023
在另一些实施方案中，所述赖氨酸类似物具有如3所示结构：
Figure PCTCN2021089174-appb-000024
其中a为1或者2的整数；b和c相同或不同且为1至4的整数。
在另一些实施方案中，所述赖氨酸类似物具有如4所示结构：
Figure PCTCN2021089174-appb-000025
其中a为0至2的整数；b和c相同或不同且为2至6的整数。
在另一些实施方案中，所述赖氨酸类似物具有如5所示结构：
Figure PCTCN2021089174-appb-000026
其中a为0至5的整数；b和c相同或不同且为1至5的整数。
在另一些实施方案中，所述赖氨酸类似物具有如6所示结构：
Figure PCTCN2021089174-appb-000027
其中a为0至5的整数；b和c相同或不同且为0至5的整数。
在另一些实施方案中，所述赖氨酸类似物具有如7所示结构：
Figure PCTCN2021089174-appb-000028
其中a为0至5的整数；b和c相同或不同且为1至5的整数。
在另一些实施方案中，所述赖氨酸类似物具有如8所示结构：
Figure PCTCN2021089174-appb-000029
其中a为0至5的整数；b、c和d相同或不同且为1至5的整数。
在另一些实施方案中，所述赖氨酸类似物具有如9所示结构：
Figure PCTCN2021089174-appb-000030
其中a为0至5的整数；b和c相同或不同且为1至5的整数。
本公开所述的树枝状聚合物，尤其是聚赖氨酸或聚赖氨酸类似物的核心可选自二苯甲基胺(BHA)、赖氨酸的二苯甲基胺(BHALys)或赖氨酸类似物或者如下10-26所示的结构：
Figure PCTCN2021089174-appb-000031
其中a为1至9，优选1至5的整数；
Figure PCTCN2021089174-appb-000032
其中a、b和c相同或不同且为1-5的整数，d为0-100，优选1-30的整数；
Figure PCTCN2021089174-appb-000033
其中a和b相同或不同，且为0至5的整数；
Figure PCTCN2021089174-appb-000034
其中a和c相同或不同且为1-6的整数，b为0-6的整数；
Figure PCTCN2021089174-appb-000035
其中a和d相同或不同且为1-6的整数，b和c相同或不同且为0-6的整数；
Figure PCTCN2021089174-appb-000036
其中a和b相同或不同且为1-5，特别是1-3，尤其是1的整数；
Figure PCTCN2021089174-appb-000037
其中a、b和c相同或不同，且为选自1-6的整数；
Figure PCTCN2021089174-appb-000038
其中a、b和c相同或不同，且为选自0-6的整数；
Figure PCTCN2021089174-appb-000039
其中a、b和c相同或不同，且为选自0-6的整数；
Figure PCTCN2021089174-appb-000040
其中a、b和c相同或不同，且为0-6的整数，d、e和f相同或不同，且为1-6的整数；
Figure PCTCN2021089174-appb-000041
其中a、b和c相同或不同，为1-6的整数；
Figure PCTCN2021089174-appb-000042
其中a、b、c和d可以相同或不同，且为0-6的整数；
Figure PCTCN2021089174-appb-000043
其中a、b、c和d可以相同或不同，且为1-6的整数；或者
Figure PCTCN2021089174-appb-000044
其中a、b、c和d可以相同或不同，且为0-6的整数，e、f、g和h可以相同或不同，且为1-6的整数。
本公开还涉及一种药物组合物，包含本公开的大分子和药学上可接受的载体。 在一些实施方案中，所述的药物组合物中不含增溶赋形剂，例如聚乙氧基蓖麻油、聚山梨醇酯。在一些实施方案中，所述的药物组合物通过透皮、口服、注射等方式给药。
本公开的大分子可以被配制成的组合物，其包括适于经口、直肠、局部、鼻内、吸入、气雾剂、眼部、或肠胃外(包括腹膜内、静脉内、皮下或肌内注射)施用的那些。组合物可方便地以单位剂型呈现并且可通过制药领域众所周知的任何方法制备。所述方法包括使大分子与构成一种或多种辅助成分的载体缔合的步骤。通常，组合物通过使大分子与液体载体缔合形成溶液或悬浮液制备，或可选地使大分子与适于形成固体，任选地颗粒产物的制剂组分缔合，然后，如果必要，将产物成型为所需的递送形式。本发明的固体制剂，当为颗粒时，通常将包含约1纳米至约500微米的颗粒尺寸范围。通常，针对预期用于静脉内施用的固体制剂，颗粒直径范围通常将为约1nm至约10微米。组合物可含有本发明的大分子，所述大分子为具有小于1000nm，例如5至1000nm，特别是5至500nm，尤其是5至400nm(如5至50nm且特别是5至20nm)的颗粒直径的纳米颗粒。在特定实施方案中，组合物含有具有5至20nm的平均尺寸的大分子。在一些实施方案中，大分子在组合物中为多分散的，PDI为1.01至1.8之间，特别是1.01至1.5之间，且尤其是1.01至1.2之间。在特定实施方案中，大分子在组合物中为单分散的。特别优选的为无菌的、冻干的组合物，所述组合物在注射前于水性媒介物中重构。
在一些实施方案中，组合物含有具有5至20nm的平均尺寸的大分子。在一些实施方案中，所述大分子的颗粒尺寸D 90或者D 50小于1000nm，例如5至1000nm，特别是5至500nm，尤其是5至400nm(如5至50nm，特别是5至20nm)。在特定实施方案中，组合物含有具有D 50为5至20nm的大分子。
本公开的大分子也可用于提供药物活性剂的控释和/或缓释制剂形式。在缓释制剂中，选择制剂成分以在延长的时期里(如数天、数周或数月)从制剂中释放大分子。这类制剂包括经皮贴剂或在可皮下沉积的可植入装置中或通过静脉注射、皮下注射、肌内注射、硬膜内注射或颅内注射。在控释制剂中，选择二酸连接子以在给定的时间窗口中释放其大部分的药物活性剂。例如，当已知大部分的大分子在靶器官、组织或肿瘤中积聚所花费的时间时，在积聚时间已过去之后可以选择连接子以释放其大部分的药物活性剂。这可允许在给定时间点在需要其作用的位点递送高载药量。可选地，选择连接子以在延长的时期里于治疗水平释放药物活性剂。在一些实施方案中，制剂可具有多种控释特性。例如，制剂包含其中药物通过不同的连接子连接的大分子，这允许快释药物的突释然后在延长的时期里以较低但恒定的治疗水平较慢释放。在一些实施方案中，制剂可具有缓释和控释特性。例如，可以选择制剂成分以在延长的时期里释放大分子且选择连接子以递送恒定的低治疗水平的药物活性剂。在一些实施方案中，药物活性剂通过不同的连接子连接于相同的分子。在一些实施方案中，每个药物-连接子组合连接于相同制 剂中的不同的大分子。
在一些实施方案中，所述的药物组合物中，大分子被配制为在5分钟至60分钟之间释放大于50％的药学活性剂。在一些实施方案中，所述的药物组合物中，大分子被配制为在2小时至48小时之间释放大于50％的药学活性剂。在一些实施方案中，所述的药物组合物中，大分子被配制为在5天至30天之间释放大于50％的药学活性剂。
本公开另一方面提供治疗或抑制肿瘤生长的方法，其包括施用有效量的本发明的大分子或含有其的药物组合物，其中作为第一端基的药物活性剂为肿瘤药物。本公开中所述的肿瘤选自黑色素瘤、脑瘤、食管癌、胃癌、肝癌、胰腺癌、结肠直肠癌、肺癌、肾癌、乳腺癌、卵巢癌、前列腺癌、皮肤癌、神经母细胞瘤、肉瘤、骨软骨瘤、骨瘤、骨肉瘤、精原细胞瘤、睾丸肿瘤、子宫癌、头颈肿瘤、多发性骨髓瘤、恶性淋巴瘤、真性红细胞增多症、白血病、甲状腺肿瘤、输尿管肿瘤、膀胱癌、胆囊癌、胆管癌、绒毛膜上皮癌和儿科肿瘤(尤因家族性肉瘤、维尔姆斯肉瘤、横纹肌肉瘤、血管肉瘤、胚胎睾丸癌、成神经细胞瘤、视网膜母细胞瘤、肝胚细胞瘤、肾母细胞瘤等)。
在本公开另一方面，提供用肿瘤药物治疗后降低过敏症的方法，其包括施用本公开的大分子或含有其的药物组合物，其中所述组合物基本上不含增溶赋形剂如Cremophor EL和聚山梨醇酯80。
在本公开的另一方面，提供降低肿瘤药物或肿瘤药物的制剂的毒性的方法，其包括施用本公开的大分子或含有其的药物组合物，其中所述肿瘤药物为大分子的第一端基。在一些实施方案中，降低的毒性为血液毒性、神经毒性、胃肠毒性、心脏毒性、肝毒性、肾毒性、耳毒性或脑病毒性。
在本公开的另一方面，提供降低与肿瘤药物或肿瘤药物的制剂相关的副作用的方法，其包括施用本公开的大分子或含有其的药物组合物，其中所述肿瘤药物为大分子的第一端基。在一些实施方案中，减少的副作用选自嗜中性白血球减少、白细胞减少、血小板减少、骨髓中毒性、骨髓抑制、神经病、疲劳、非特异性神经认知问题、眩晕、脑病、贫血、味觉障碍、呼吸困难、便秘、厌食、甲病症、液体潴留、虚弱、疼痛、恶心、呕吐、粘膜炎、秃发、皮肤反应、肌痛、过敏症和过敏性。
在一些实施方案中，本公开的大分子或包含大分子的药物组合物可以减少或消除具有试剂如皮质类固醇和抗组胺剂的术前用药的需要。
制备树枝状聚合物的方法在本领域为已知的。例如，大分子的树枝状聚合物可通过发散方法或收敛方法或其混合制备。
在发散方法中，将结构单元的每代依序加入至核心或前一代。保护具有一个或两个表面氨基的表面代。如果氨基中的一者得到保护，那么游离的氨基与连接子、连接子-药物活性剂或药代动力学修饰剂中的一者反应。如果两个氨基均得到 保护，它们以不同的保护基团得到保护，那么可以去除一个保护基团而不去除另一个保护基团。去除氨基保护基团中的一者并与连接子、连接子-药物活性剂或药代动力学修饰剂中的一者反应。一旦初始端基已连接于树枝状聚合物，那么去除另一氨基保护基团且加入另一第一和第二端基。这些基团通过本领域已知的酰胺形成连接于表面氨基。
在收敛方法中，结构单元的每一代在前一代上建立以形成树枝化基元。在树枝化基元连接于核心之前或之后，第一和第二端基可如以上所述连接于表面氨基。
在混合方法中，将结构单元的每一代加入至核心或结构单元的前一代。然而，在最后一代加入至树枝状聚合物之前，将表面氨基与端基(例如，第一和第二端基，第一和第三端基或第二和第三端基)官能化。然后将官能化的末代加入至结构单元的亚表面层并将树枝化基元连接至核心。
药物活性剂通过本领域已知的酯化反应与连接子的羧酸中的一者反应。例如，形成活化的羧酸如使用酰氯或酸酐，且与药物活性剂的羟基反应。如果药物活性剂具有一个以上的羟基，那么其它的羟基可以得到保护。
在靶向试剂连接于核心的情况下，核心上的官能团可在树枝状聚合物形成期间得到保护然后去保护并与靶向试剂、连接基团或靶向试剂-连接基团反应。可选地，在形成树枝状聚合物之前，核心可与连接基团或靶向试剂-连接基团反应。
合适的保护基团、其引入和去除的方法描述于Greene&Wuts,Protecting Groupsin Organic Synthesis，第三版，1999。
本公开还包括大分子或其药学上可接受的盐的各种氘代形式，大分子中的每个可用的氢原子可独立地被氘原子置换。本领域技术人员将知晓如何合成本公开的大分子或其药学上可接受的盐的氘代形式。
本公开还包括同位素标记的大分子，大分子中的一个或多个原子被具有不同于自然界中最常见的原子质量或质量数的原子替换。可用于本公开的大分子的同位素的实例包括氢、碳、氮、氧、氟、碘和氯的同位素，例如 3H、 11C、 14C、 18F、 123I或 125I。
本公开的大分子可以呈药学上可接受的盐的形式。但是，应理解非药学上可接受的盐也落入本公开的范围内，因为这些可以在药学上可接受的盐的制备中用作中间体或在储存或运输过程中可能为有用的。合适的药学上可接受的盐包括但不限于药学上可接受的无机酸(如盐酸、硫酸、磷酸、硝酸、碳酸、硼酸、氨基磺酸和氢溴酸)的盐，或药学上可接受的有机酸(如乙酸、丙酸、丁酸、酒石酸、马来酸、羟基马来酸、富马酸、马来酸、柠檬酸、乳酸、黏酸、葡萄糖酸、苯甲酸、琥珀酸、草酸、苯乙酸、甲烷磺酸、甲苯磺酸(toluenesulphonic)、苯磺酸、水杨酸、磺胺酸(sulphanilic)、天冬氨酸、谷氨酸、乙二胺四乙酸、硬脂酸、棕榈酸、油酸、月桂酸、泛酸、鞣酸、抗坏血酸和戊酸)的盐。碱式盐包括但不限于与药学上可接受的阳离子(如钠、钾、锂、钙、镁、铵和烷基铵)形成的那些。
除非有相反陈述，否则下列用在说明书和权利要求书中的术语具有下述含义。
本公开所述的“被(一个或多个)氧、硫或氮原子间断”，例如“被(一个或多个)氧、硫或氮原子间断的C 1-10亚烷基”，指氧、硫或氮原子可以位于烷基链内部、也可以在烷基链的末端。例如，
Figure PCTCN2021089174-appb-000045
表示连接子-X 1-L-X 2-中，X 1不存在，并且X 2为-C(O)-，与树枝状聚合物D的表面氨基连接形成酰胺键，该处的亚烷基被氧和硫间断，硫位于烷基链的内部，氧在烷基链的末端。另外，本公开为了方便对连接子以及连接子与树枝状聚合物、药学活性剂的连接关系进行描述引入了“D”和“A”，连接子本身并不包含“D”、“A”、或者“D”、“A”上的基团。
术语“烷基”是指具有1至10个碳原子的直链或支链饱和烃基。在适当的情况下，烷基可具有指定的碳原子数，例如，C 1-4烷基，所述烷基包括在直链或支链排列中具有1、2、3或4个碳原子的烷基。合适的烷基的实例包括但不限于甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、正戊基、2-甲基丁基、3-甲基丁基、4-甲基丁基、正己基、2-甲基戊基、3-甲基戊基、4-甲基戊基、5-甲基戊基、2-乙基丁基、3-乙基丁基、庚基、辛基、壬基和癸基。
术语“亚烷基”是指具有1至10个碳原子的直链或支链二价烷基。
术语“烯基”是指包括具有2至12个碳原子的支链和直链烯烃或含有脂族烃基团的烯烃。例如“C 2-6烯基”表示具有2、3、4、5或6个碳原子的烯基。烯基的实例包括但不限于，乙烯基、烯丙基、1-丙烯基、1-丁烯基、2-丁烯基、3-丁烯基、2-甲基丁-2-烯基、3-甲基丁-1-烯基、1-戊烯基、3-戊烯基及4-己烯基。
术语“炔基”是指包括具有2至12个碳原子的支链和直链炔基或含有脂族烃基的烯烃，或若规定指定碳原子数，则意指该特定数目。例如乙炔基、丙炔基(例如1-丙炔基、2-丙炔基)、3-丁炔基、戊炔基、己炔基及1-甲基戊-2-炔基。
“亚烯基”、“亚炔基”是指衍生自烯基或炔基的部分不饱和的支链或直链二价烃基。在某些实施例中，此类亚烯基任选地被取代。亚烯基的非限制性实例包括亚乙烯基、亚丙烯基、亚丁烯基、亚戊烯基、亚己烯基、亚庚烯基、亚辛烯基、亚壬烯基、亚癸烯基及类似基团；亚炔基的非限制性实例包括亚乙炔基、亚丙炔基、亚丁炔基、亚戊炔基、亚己炔基及类似基团。
“环烷基”是指饱和或部分不饱和的环烃。环烷基环可包括指定的碳原子数。例如，3至8元环烷基包括3、4、5、6、7或8个碳原子。合适的环烷基的实例包括但不限于环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、1,4-环己二烯基、环庚基和环辛基。
术语“亚环烷基”是指衍生自环烷基的二价环烃基。例如
Figure PCTCN2021089174-appb-000046
等。
术语“烷氧基”是指-O-(烷基)和-O-(非取代的环烷基)，其中烷基的定义如上所述。烷氧基的非限制性实例包括：甲氧基、乙氧基、丙氧基、丁氧基、环丙氧基、环丁氧基、环戊氧基、环己氧基。烷氧基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自H原子、D原子、卤素、烷基、烷氧基、卤代烷基、羟基、羟烷基、氰基、氨基、硝基、环烷基、杂环基、芳基、杂芳基中的一个或多个取代基所取代。
术语“芳基”是指在各个环中至多7个原子的任何稳定的、单环或双环碳环，其中至少一个环为芳族的。此类芳基的实例包括但不限于苯基、萘基、四氢萘基、茚满基、联苯基或联萘基。
术语“杂环烷基”或“杂环基”是指环烃，其中一至四个碳原子已被独立地选自N、N(R)，S，S(O)、S(O) 2和O的杂原子替换。杂环可为饱和的或不饱和的。合适的杂环基的实例包括四氢呋喃基、四氢噻吩基、吡咯烷基、吡咯啉基、吡唑啉基、吡喃基、哌啶基、吡唑啉基、二硫杂环戊二烯基、氧杂硫杂环戊二烯基、二氧杂环己基、二氧杂环己烯基、吗啉代和噁嗪基。
术语“杂芳基”表示在各个环中至多7个原子的稳定的单环或双环，其中至少一个环为芳族且至少一个环含有选自O、N和S的1至4个杂原子。在本定义范围内的杂芳基包括但不限于吖啶基、咔唑基、噌啉基、喹喔啉基、喹唑啉基、吡唑基、吲哚基、苯并三唑基、呋喃基、噻吩基、苯硫基、3,4-亚丙基二氧苯硫基(3,4-propylenedioxythiophenyl)、苯并噻吩基、苯并呋喃基、苯并二噁烷、苯并二氧杂环己烯、喹啉基、异喹啉基、噁唑基、异噁唑基、咪唑基、吡嗪基、哒嗪基、吡啶基、嘧啶基、吡咯基、四氢喹啉、噻唑基、异噻唑基、1,2,4-三唑基、1,2,3-三唑基、1,2,4-噁二唑基、1,2,4-噻二唑基、1,3,5-三嗪基、1,2,4三嗪基、1,2,4,5四嗪基和四唑基。
术语“卤素”是指氟、氯、溴或碘。
术语“卤代”是指被一个或多个选自氟、氯、溴、碘的原子取代。
“树枝状聚合物”是指含有核心和至少一个连接于所述核心的树枝化基元的分子。每个树枝化基元由至少一层或一代分支的结构单元构成，产生具有每代结构单元的分支的越来越多的分支结构。连接于核心的树枝化基元的最大数目受限于核心上官能团的数目。
“结构单元”是指具有至少三个官能团的分支的分子，一个官能团用于连接于核心或结构单元的上一代且至少两个官能团用于连接于结构单元的下一代或形成树枝状聚合物的表面。
“代”是指构成树枝化基元或树枝状聚合物的结构单元的层数。例如，一代树枝状聚合物将具有一层连接于所述核心的分支的结构单元，例如，核心-[[结构单元]] u，其中u为连接于核心的树枝化基元的数目。两代树枝状聚合物在连接于核心的每个树枝化基元中具有两层结构单元，当所述结构单元具有一个分支点时， 所述树枝状聚合物可为：核心[[结构单元][结构单元] 2] u。三代树枝状聚合物在连接于核心的每个树枝化基元中具有三层结构单元，例如核心-[[结构单元][结构单元] 2[结构单元] 4]u。六代树枝状聚合物具有六层连接于核心的结构单元，例如核心-[[结构单元][结构单元] 2[结构单元] 4[结构单元] 8[结构单元] 16[结构单元] 32] u等。结构单元的最后一代(最外代)提供树枝状聚合物的表面官能化和可用于结合端基的官能化基团的数目。例如，在具有两个树枝化基元连接于核心的树枝状聚合物中(u＝2)，如果每个结构单元具有一个分支点并且存在6代时，那么最外代具有64个结构单元和128个可用于结合端基的官能团。
“微溶的”是指在水中具有1mg/mL至10mg/mL的溶解度的药物或药物活性剂。在水中溶解度小于1mg/mL的药物被认为是不溶的。
“增溶赋形剂”是指用于将不溶的或微溶的药学活性剂溶解于水性制剂的制剂添加剂。实例包括表面活性剂如聚乙氧基蓖麻油包括Cremophor EL、CremophorRH40和Cremophor RH60，D-α生育酚-聚乙二醇1000琥珀酸盐、聚山梨醇酯20、聚山梨醇酯80、solutol HS15、山梨醇单油酸酯(sorbitan monoleate)、泊洛沙姆407、Labrasol等。
“任选的”或“任选”是指意味着随后所描述的事件或环境可以但不必发生，该说明包括该事件或环境发生或不发生的场合。例如，“L为任选被一个或多个氧、硫或氮原子间断的C 1-10直链亚烷基”意味着C 1-10直链亚烷基可以被氧、硫或氮原子间断但不必须被间断，该说明包括C 1-10直链亚烷基被氧、硫或氮原子间断的情形和C 1-10直链亚烷基不被氧、硫或氮原子间断的情形。
“取代的”指基团中的一个或多个氢原子，优选为最多5个，更优选为1～3个氢原子彼此独立地被相应数目的取代基取代。不言而喻，取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下(通过实验或理论)确定可能或不可能的取代。
本公开所述化合物的化学结构中，键
Figure PCTCN2021089174-appb-000047
并未指定构型，即键
Figure PCTCN2021089174-appb-000048
可以为
Figure PCTCN2021089174-appb-000049
或者同时包含
Figure PCTCN2021089174-appb-000050
两种构型。本公开所述化合物的化学结构中，键
Figure PCTCN2021089174-appb-000051
并未指定构型，即可以为Z构型或E构型，或者同时包含两种构型。
本公开所述化合物或其可药用盐、或其异构体的任何同位素标记的衍生物都被本公开所覆盖。能够被同位素标记的原子包括但不限于氢、碳、氮、氧、磷、氟、氯、碘等。它们可分别被同位素 2H(D)、 3H、 11C、 13C、 14C、 15N、 18F、 31P、 32P、 35S、 36Cl和 125I等代替。除另有说明，当一个位置被特别地指定为氘(D)时，该位置应理解为具有大于氘的天然丰度(其为0.015％)至少3000倍的丰度的氘(即，至少45％的氘掺入)。
本公开中药学活性剂或其残基、药学活性剂、药学活性剂或其残基A可互换使用，均是指具有药学活性的分子或者基团。
附图说明
图1为测试例1中本公开化合物的人血浆转化浓度曲线。
图2为测试例2中的比格犬血浆PK曲线。
具体实施方式
以下结合实施例进一步描述解释本发明，但这些实施例并非意味着限制本发明的范围。
本发明实施例中未注明具体条件的实验方法，通常按照常规条件，或按照原料或商品制造厂商所建议的条件。未注明具体来源的试剂，为市场购买的常规试剂。
化合物的结构是通过核磁共振(NMR)或质谱(MS)来确定的。NMR的测定是用Bruker AVANCE-400核磁仪，测定溶剂为氘代二甲基亚砜(DMSO-d6)、氘代氯仿(CDCl 3)、氘代甲醇(CD 3OD)，内标为四甲基硅烷(TMS)，化学位移是以10 -6(ppm)作为单位给出。
MS的测定用FINNIGAN LCQAd(ESI)质谱仪(生产商：Thermo,型号：Finnigan LCQ advantage MAX)。
高效液相色谱法(HPLC)的测定使用安捷伦1200DAD高压液相色谱仪(Sunfire C18 150×4.6mm色谱柱)和Waters 2695-2996高压液相色谱仪(Gimini C18 150×4.6mm色谱柱)。
薄层层析硅胶板使用烟台黄海HSGF254或青岛GF254硅胶板，薄层色谱法(TLC)使用的硅胶板采用的规格是0.15mm～0.2mm，薄层层析分离纯化产品采用的规格是0.4mm～0.5mm硅胶板。
柱层析一般使用烟台黄海200～300目硅胶为载体。
本发明的已知的起始原料可以采用或按照本领域已知的方法来合成，或可购买自ABCR GmbH&Co.KG，Acros Organnics，Aldrich Chemical Company，韶远化学科技(Accela ChemBio Inc)、达瑞化学品等公司。
实施例中如无特殊说明，反应均在氩气氛或氮气氛下进行。
氩气氛或氮气氛是指反应瓶连接一个约1L容积的氩气或氮气气球。
氢气氛是指反应瓶连接一个约1L容积的氢气气球。
加压氢化反应使用Parr 3916EKX型氢化仪和清蓝QL-500型氢气发生器或HC2-SS型氢化仪。
氢化反应通常抽真空，充入氢气，反复操作3次。
微波反应使用CEM Discover-S 908860型微波反应器。
实施例中如无特殊说明，反应中的溶液是指水溶液。
实施例中如无特殊说明，反应的温度为室温。
室温为最适宜的反应温度，温度范围是20℃～30℃。
实施例中pH＝6.5的PBS缓冲液的配制：取KH 2PO 4 8.5g，K 2HPO 4.3H 2O 8.56g，NaCl 5.85g，EDTA 1.5g置于瓶中，定容至2L，超声波使其全部溶解，摇匀即得。
纯化化合物采用的柱层析的洗脱剂的体系和薄层色谱法的展开剂的体系包括：A：二氯甲烷和异丙醇体系，B：二氯甲烷和甲醇体系，C：石油醚和乙酸乙酯体系，溶剂的体积比根据化合物的极性不同而进行调节，也可以加入少量的三乙胺和酸性或碱性试剂等进行调节。
本披露部分化合物是通过Q-TOF LC/MS来表征的。Q-TOF LC/MS使用安捷伦6530精确质量数四级杆-飞行时间质谱仪和安捷伦1290-Infinity超高效液相色谱仪(安捷伦Poroshell 300SB-C8 5μm,2.1×75mm色谱柱)。
参考专利CN 110312531A的合成方法，合成得到了如下一类树枝状聚合物，其第一氨基端基用于与药物活性剂的连接，第二端基用于与药代动力学修饰剂PEG相连：
Figure PCTCN2021089174-appb-000052
在下文实施例中表示的树枝状聚合物包括参考树枝状聚合物的核心和树枝状聚合物最外代中的结构单元。未描绘第1代至亚表面代。树枝状聚合物BHALys[Lys] 32表示5代树枝状聚合物，其具有下式BHALys[Lys] 2[Lys] 4[Lys] 8[Lys] 16[Lys] 32，64个表面氨基可用于结合于端基。
树枝状聚合物支架BHALys[Lys] 32[α-NH 2·TFA] 32[ε-PEG 570] 32、BHALys[Lys] 32[α-NH 2·TFA] 32[ε-PEG 1100] 32、BHALys[Lys] 32[α-NH 2·TFA] 32[ε-t-PEG 2300] 32、BHALys[Lys] 32[α-4-HSBA] 32[ε-PEG 1100] 32、BHALys[Lys] 32[α-GILGVP-NH 2.TFA] 32[ε-PEG 1100] 32和BHALys[Lys] 32[α-GILGVP-NH 2·TFA] 32[ε-t-PEG 2300] 32的制备可见于Kaminskas等人，J Control.Release(2011)(doi：10.1016/j.jconrel.2011.02.005)。树枝状聚合物支架4-叠氮苯甲酰胺-PEG 12-NEOEOEN[Su(NPN) 2][Lys] 16[NH 2·TFA] 32的制备可见于WO08/017122。
1-A00、1-B00和1-C00是根据专利WO2012167309A合成，其连接的药学活性剂为多西他赛；2-A00是根据专利WO2018154004A合成，其连接的药学活性剂为化合物2。树枝状聚合物支架BHALys[Lys] 32[α-NH 2·TFA] 32[ε-PEG 1100] 32在实施例中记为Dendrimer 1，BHALys[Lys] 32[α-NH 2·TFA] 32[ε-PEG 2100] 32在实施例中记为Dendrimer 1-PEG2K，根据WO2018154004A的方法合成：
Figure PCTCN2021089174-appb-000053
一般程序
一般程序A
连接子与药物的安装
在0℃，向磁力搅拌的羧酸连接子(0.2–0.5mmol)在溶剂DMF或乙腈(1–5mL)的溶液中加入EDC或DCC(1.2当量)的偶联剂。将混合物放置搅拌5分钟，然后滴加含有药物(0.4–1当量)和DMAP(0.4–1当量)的混合物的溶液(1mL)。将混合物在0℃保持1小时，然后使其升温至环境温度。然后真空去除挥发物并将残余物 通过制备型HPLC(BEH300Waters XBridge C18，5μM，30x150mm，40-80％ACN/水(5-40min)，不含缓冲液)纯化以获得所需产物。
一般程序B
连接子与药物的安装
向磁力搅拌的药物(0.3–1.0mmol)和酸酐(2当量)在DMF(3-5mL)的溶液中加入DIPEA(3当量)。将混合物在环境温度搅拌过夜。然后真空去除挥发物并将残余物通过制备型HPLC(BEH300Waters XBridge C18，5μM，30x150mm，40-70％ACN/水(5-40min)，不含缓冲液,RT＝34min)纯化。将适当的级分真空浓缩以得到所需靶物。
一般程序C
具有药物-连接子负载的树枝状聚合物。
在室温，向磁力搅拌的BHALys[Lys] 32[α-NH 2·TFA] 32[ε-PEG 1100] 32(0.5–1.0μmol)和DIPEA(1.2当量/胺)在DMF的混合物中加入连接子-药物(1.2当量/胺基)和PyBOP(1.2当量/胺基)。在室温搅拌1.5小时之后，去除挥发物并将残余物通过SEC(sephadex，LH20，MeOH)纯化。将适当的级分(如通过HPLC判断)合并，并浓缩以得到所需物质。
一般程序D
点击反应
向磁力搅拌的以1:1H 2O/t-BuOH(大约0.5mL)的树枝状聚合物溶液(0.5–1.0mmol)中，加入炔烃试剂(2当量)、抗坏血酸钠溶液(2当量)和CuSO 4溶液(20mol％)。将溶液加热至80℃并用HPLC监测。根据需要加入额外量的抗坏血酸钠和CuSO 4以驱动反应完成。在判断反应完成之后，真空浓缩反应物，然后纯化。
本公开中大分子可按照选自以下路线进行合成，PG为羧酸保护基：
Figure PCTCN2021089174-appb-000054
Figure PCTCN2021089174-appb-000055
实施例1：本公开化合物1-F00的制备
Figure PCTCN2021089174-appb-000056
Figure PCTCN2021089174-appb-000057
第一步
氮气氛下，将1-F01-2(44g，0.23mol，按照专利US2005020645A合成)和溴乙酸叔丁酯(53.6g，0.28mol)溶于二甲基亚砜(640mL)中，冷却至0℃，加入KOH(25.8g，0.46mol)，于0℃下搅拌1小时。加入1N盐酸中和，用乙酸乙酯(200mL×3)萃取，浓缩后经硅胶柱层析纯化得44g产物1-F01-3，收率69％。
第二步
将化合物1-F01-3(3.1g，10.13mmol)溶于二氯甲烷(16mL)中，加入三氟乙酸(8mL)，室温搅拌3小时。浓缩后加入甲基叔丁基醚(MTBE)(20mL)溶解，用饱和碳酸氢钠溶液中和，水相用MTBE(10mL)洗涤，用1N盐酸调pH至2，用乙酸乙酯萃取，浓缩后得2.1g产物1-F01-4，收率71％。
1H NMR(400MHz,DMSO-d 6)δ12.64(br,1H),7.33-7.41(m,5H),5.15(s,2H),4.16(s,2H),1.18-1.30(m,4H)。
第三步
将化合物多西他赛(DTX)(360mg，0.45mmol)、1-F01-4(145mg，0.58mmol)和二甲氨基吡啶(DMAP)(55mg，0.45mmol)溶于二氯甲烷(5mL)中，冷却至0℃，滴加二环己基碳二亚胺(DCC)(120mg，0.58mmol)的二氯甲烷(1mL) 溶液，室温搅拌过夜。浓缩后通过高效液相色谱法制备得332mg产物1-F01-Bn，收率71％。
MS-ESI：m/z 1040.4[M+H] +。
第四步
将化合物1-F01-Bn(300mg，0.29mmol)溶于四氢呋喃(5mL)中，加入Pd/C(50mg，10wt.％)，在氢气氛下，于室温搅拌过夜。过滤，滤液减压浓缩，得270mg产物1-F01，收率99％。MS-ESI:m/z 950.3[M+H] +。
第五步
氮气氛下，将化合物1-F01(57mg，60.4μmol)与六氟磷酸苯并三唑-1-基-氧基三吡咯烷基磷(PyBOP)(60mg，115.5μmol)溶于无水N,N-二甲基甲酰胺(DMF)(2mL)中，滴加化合物Dendrimer 1(68mg，1.5μmol)与二异丙基乙基胺(DIPEA)(15mg，115.5μmol)的无水DMF(2mL)混合溶液，反应1.5小时。将反应液直接通过柱层析色谱法纯化，得到72mg油状化合物。用纯化水(20mL)溶解，经滤膜(0.22μm)过滤，冻干后得到70mg产物1-F00，收率79％。
1H NMR指示19DTX/树枝状聚合物。实际的分子量大约为59.1kDa(25.9％重量％DTX)。
1H NMR(400MHz,CD 3OD)δ8.02-8.25(m,59H),7.12-7.94(m,261H),6.00-6.31(m,19H),5.54-5.72(m,22H),5.20-5.51(m,86H),4.96-5.08(m,45H),4.10-4.68(m,138H),3.40-4.04(m,3100H),3.33-3.36(s,91H),3.02-3.23(m,131H),2.22-2.65(m,174H),1.00-2.10(m,1362H)。
实施例2：本公开化合物1-L00的制备
Figure PCTCN2021089174-appb-000058
Figure PCTCN2021089174-appb-000059
第一步
将化合物1-L01-6A(246.7mg，1.56mmol，根据文献Organic Letters,2006,8,5685–5688合成)、多西他赛(840mg，1.04mmol)和DCC(385.6mg，1.87mmol)溶于DMF(15mL)中，加入DMAP(190.6mg，1.56mmol)，室温搅拌16小时。HPLC制备纯化，得到370mg产物1-L01，收率37.5％。
MS-ESI：m/z 948.4[M+H] +。
第二步
氮气氛下，将化合物1-L01(30.3mg，31.9μmol)和PyBOP(31.9mg，61.5μmol)溶于无水DMF(1.0mL)中，滴加Dendrimer 1(36mg，0.799μmol)与DIPEA(7.9mg，61.5μmol)的无水DMF(1.0mL)混合溶液，反应1.5小时。将反应液直接通过柱层析色谱法纯化，得到37mg油状化合物。用纯化水(10mL)溶解，经滤膜(0.22μm)过滤，冻干后得到35mg产物1-L00，收率72％。
1H NMR指示21DTX/树枝状聚合物。实际的分子量大约为60.9kDa(27.8％重量％DTX)。
1H NMR(400MHz,CD 3OD)δ8.02-8.25(m,58H),7.12-7.94(m,263H),6.00-6.30(m,21H),5.53-5.76(m,25H),5.11-5.46(m,113H),4.96-5.10(m,70H),4.05-4.52(m,77H),3.40-3.96(m,3100H),3.33-3.36(S,103H),2.97-3.23(m,130H),2.25-2.74(m,216H),1.00-2.10(m,1373H)。
实施例3：本公开化合物1-M00的制备
Figure PCTCN2021089174-appb-000060
第一步
将化合物1-1(711mg，5mmol)溶于四氢呋喃(10mL)中，冷却至-70℃，滴加二异丙基氨基锂(LDA)(3.5mL，7mmol)。搅拌30分钟后，加入化合物1-2(551mg，5.5mmol)，继续搅拌2小时。加水淬灭，用2N盐酸调pH为6～7，用乙酸乙酯萃取，浓缩后得到700mg粗品1-M01-a，直接用于下一步反应。
第二步
将化合物1-M01-a(700mg，2.89mmol)溶解于甲醇(100mL)中，加入KOH (162mg，2.89mmol)，于室温搅拌16小时。加水稀释，用2N盐酸调pH为3～5，用乙酸乙酯萃取，浓缩后得到650mg粗品1-M01-b，直接用于下一步反应。
第三步
将化合物1-M01-b(650mg，3.03mmol)、溴化苄(519mg，3.03mmol)和碳酸钾(836mg，6.06mmol)加入到DMF(10mL)中，于室温搅拌3小时。加水稀释后，用乙酸乙酯萃取，浓缩后得800mg粗品1-M01-c，直接用于下一步反应。
第四步
将化合物1-M01-c(800mg，2.63mmol)溶于二氯甲烷(5mL)中，加入三氟乙酸(10mL)，于室温搅拌2小时。浓缩后HPLC制备纯化得60mg产品1-M01-d，四步收率6％。
MS-ESI：m/z 249.2[M+H] +。
第五步
将化合物1-M01-d(37mg，0.1485mmol)、多西他赛(60mg，0.0743mmol)和DCC(27.6mg，0.1337mmol)溶于DMF(15mL)中，加入DMAP(13.6mg，0.1114mmol)，于室温搅拌16小时。HPLC制备纯化，得到15mg 1-M01-Bn，收率19.5％。
MS-ESI：m/z 1038.4[M+H] +。
第六步
将化合物1-M01-Bn(14mg，0.0135mmol)溶于四氢呋喃(3mL)中，加入Pd/C(3mg，10wt.％)，氢气氛下，室温搅拌24小时。HPLC制备纯化得10mg产品1-M01，收率78％。
MS-ESI：m/z 948.4[M+H] +。
第七步
氮气氛下，将化合物1-M01(50.4mg，53.2μmol)和PyBOP(53.2mg，102.3μmol)溶于无水DMF(2mL)中，滴加化合物Dendrimer 1(60mg，1.33μmol)与DIPEA(13.2mg，102.3μmol)的无水DMF(1mL)混合溶液，反应1.5小时。将反应液直接通过柱层析色谱法纯化，得到63mg油状化合物。用纯化水(15mL)溶解，经滤膜(0.22μm)过滤，冻干后得到60mg产物1-M00，收率73％。
1H NMR指示22DTX/树枝状聚合物。实际的分子量大约为61.8kDa(28.7％重量％DTX)。
1H NMR(400MHz,CD 3OD)δ8.02-8.27(m,48H),7.13-7.94(m,204H),6.00-6.30(m,22H),5.53-5.72(m,24H),5.08-5.45(m,108H),4.10-4.68(m,141H),3.40-4.04(m,3100H),3.33-3.37(s,98H),3.06-3.23(m,109H),2.23-2.66(m,176H),1.00-2.10(m,1128H)。
实施例4：本公开化合物1-Q00的制备
Figure PCTCN2021089174-appb-000061
第一步
将化合物1-F01-3(4g，13.07mmol)溶于乙酸乙酯(80mL)中，加入Pd(OH) 2/C(0.8g，20wt.％)，氢气氛下，室温搅拌过夜。过滤，滤液减压浓缩，得2.6g产物1-F01a，收率92％。
1H NMR(400MHz,DMSO-d 6)δ4.13(s,2H),1.49(s,9H),1.43-1.47(m,2H),1.30-1.33(m,2H)。
第二步
将化合物1-F01a(2.6g，12.03mmol)溶于DMF(60mL)中，加入烯丙基溴(4.37g，36.09mmol)和NaHCO 3(2.32g，27.67mmol)，室温搅拌过夜。加热到50℃，继续反应2小时。加入1N盐酸(100mL)淬灭，用乙酸乙酯(50mL×2)萃取，浓缩后经柱层析色谱法纯化得2.1g产物1-Q01-1，收率68％。
第三步
将化合物1-Q01-1(2.1g，8.20mmol)溶于二氯甲烷(10mL)中，冷却至0℃，加入三氟乙酸(22mL)，升至室温搅拌过夜。浓缩后得1.93g粗品产物1-Q01-2，直接用于下一步反应。
第四步
氮气氛下，将化合物1-Q01-2(1.93g，9.65mmol)、苯甲醇(1.25g，11.57mmol)和DMAP(235mg，1.93mmol)溶于二氯甲烷(20mL)中，分批加入DCC(2.39g，11.57mmol)，室温搅拌过夜。浓缩后经柱层析色谱法纯化得2g产物1-Q01-3，收率71％。
第五步
氮气氛下，将化合物1-Q01-3(2g，6.89mmol)、Pd(PPh 3) 2Cl 2(73mg，0.1mmol)溶于二氯甲烷(60mL)和水(6mL)中，加入Bu 3SnH(2.95g，10.13mmol)，室温搅拌0.5小时。加水(5mL)稀释后，用二氯甲烷(10mL×2)萃取，浓缩后通过HPLC制备纯化得510mg产物1-Q01-P。
1H NMR(400MHz,DMSO-d 6)δ13.07(br,1H),7.31-7.38(m,5H),5.14(s,2H),4.31(s,2H),1.15-1.17(m,4H)。
第六步
氮气氛下，将化合物多西他赛(240mg，0.30mmol)，1-Q01-P(89mg，0.36mmol)和DMAP(37mg，0.30mmol)溶于二氯甲烷(4mL)中，冷却至0℃，滴加DCC(74mg，0.36mmol)的二氯甲烷溶液(0.5mL)，室温搅拌过夜。HPLC制备纯化，得111mg产物1-Q01-Bn，收率36％。
MS-ESI：m/z 1040.4[M+H] +。
第七步
将化合物1-Q01-Bn(110mg，0.11mmol)溶于四氢呋喃(5mL)中，加入Pd/C(30mg，10wt.％)，氢气氛下，室温搅拌过夜。过滤，滤液减压浓缩得98mg产物1-Q01，收率98％。
MS-ESI：m/z 950.3[M+H] +。
第八步
氮气氛下，将化合物1-Q01(60.6mg，63.9μmol)与PyBOP(63.6mg，122.3μmol)溶于无水DMF(1.5mL)中，滴加化合物Dendrimer 1(72mg，1.59μmol)与DIPEA(15.8mg，102.3μmol)的无水DMF(1.5mL)混合溶液，反应1.5小时。将反应液直接通过柱层析色谱法纯化，得到80mg油状化合物。用纯化水(20mL)溶解，经滤膜(0.22μm)过滤，冻干后得到78mg产物1-Q00，收率76％。
1H NMR指示25DTX/树枝状聚合物。实际的分子量大约为64.7kDa(31.2％重量％DTX)。
1H NMR(400MHz,CD 3OD)δ8.02-8.23(m,59H),7.11-7.90(m,256H),6.00-6.30 (m,25H),5.54-5.70(m,25H),5.08-5.42(m,108H),4.10-4.64(m,150H),3.40-3.96(m,3100H),3.33-3.36(s,108H),2.98-3.24(m,129H),2.28-2.62(m,176H),1.00-2.10(m,1346H).
实施例5：本公开化合物1-S00的制备
Figure PCTCN2021089174-appb-000062
第一步
将化合物1-T01-1(200.0g，1.66mol)溶于二氯甲烷(2.5L)中，加入碳酸氢钠(168.0g，2.0mol)，分批加入化合物1-T01-2B(4,4'-双甲氧基三苯甲基氯，563.2g，1.66mol)，室温搅拌19小时。加水(1000mL)淬灭，用二氯甲烷萃取，浓缩得油状物，加入乙酸乙酯(400mL)溶解，滴加石油醚(2000mL)，室温继续搅拌半小时，过滤，干燥，得到587.2g产物1-T01-3A，收率83.4％。
MS-ESI：m/z 423.2[M+H] +。
第二步
将化合物1-01-3A(400.0g，0.947mol)溶于四氢呋喃(3L)中，冷却至-60℃，滴加LDA(1.042L，2.084mol，2M在THF/己烷中)。于-60℃反应1.5小时，加入1,3,2-二氧杂噻吩烷-2,2-二氧化物(129.2g，1.042mol)，继续搅拌20分钟，加入二甲基丙烯基脲(DMPU)(133.4g，1.042mol)，搅拌1小时。升至室温搅拌19小时。加入1升10％氯化铵水溶液淬灭，用乙酸乙酯萃取，浓缩后得436.5g粗品产物1-T01-4A，收率102.8％。
MS-ESI：m/z 449.2[M+H] +。
第三步
将化合物1-T01-4A(436.5g，947mmol)溶于二氯甲烷(2.5L)中，加入三乙基硅烷(131.9g，1137mmol)，冷却至5℃，滴加三氟乙酸/二氯甲烷(74.5g的三氟乙酸溶于100mL的二氯甲烷)混合溶液，室温搅拌23小时。含1-T01-5B的反应液直接用于下一步反应。
第四步
将NaOH(227.3g,5.68mol)的水溶液(1L)滴入上一步的含1-T01-5B的反应液中，随后加入四氢呋喃(600mL)，加热至55℃，搅拌3小时。冷却至室温，减压除去有机溶剂，水相用乙酸乙酯洗涤，然后用1N稀盐酸调节pH至6得1-T01-6B水溶液，直接用于下一步反应。
第五步
向上一步得到的1-T01-6B水溶液(111.8g，947mmol)中加入四氢呋喃(300mL)，冷却至0℃，滴加溴乙酸叔丁酯(92.4g，663mmol)，然后升至室温搅拌20小时。减压除去四氢呋喃，用2N盐酸调pH至3～4，用乙酸乙酯萃取，浓缩后通过柱层析色谱法纯化，得到49.3g粗品。加入250毫升石油醚，室温搅拌2小时，过滤干燥得到25.0g产物1-T01-tBu，四步总收率11.1％。
MS-ESI：m/z 255.1[M+Na] +。
第六步
将化合物1-T01-tBu(6.0g，25.8mmol)溶于DMF(30mL)中，依次加入溴化苄(5.3g，31.0mmol)和碳酸钾(7.1g，51.7mmol)，加热至70℃，搅拌1.5小时。冷却后加水(150mL)稀释，用乙酸乙酯(100mL)萃取，浓缩得9.9 g粗品1-S01-4A。收率122.4％。
MS-ESI：m/z 345.2[M+Na] +。
第七步
将化合物1-S01-4A(10.2g，25.8mmol)溶于二氯甲烷(51mL)中，滴加三氟乙酸(51mL)，室温搅拌1小时。浓缩后经柱层析色谱法纯化得到6.2g产物1-S01-P，收率73.5％。
MS-ESI：m/z 267.2[M+H] +。
1H NMR(400MHz,CDCI 3)δ7.38-7.26(m,5H),5.14(s,2H),3.47(s,2H),1.71-1.68(m,2H),1.35-1.32(m,2H)。
第八步
将化合物多西他赛(3.0g，3.71mmol)、1-S01-P(1.087g，4.08mmol)和DMAP(0.498g，4.08mmol)溶于DMF(30mL)中，冷却至0℃，加入(1-乙基-3(3-二甲基丙胺)碳二亚胺)(EDCI)(0.783g，4.08mmol)。然后升至室温，搅拌16小时。加水(300mL)淬灭，用乙酸乙酯萃取，浓缩后经柱层析色谱法纯化，得到3.11g产物1-S01-Bn，收率77％。
MS-ESI：m/z 1078.3[M+Na] +。
第九步
将化合物1-S01-Bn(3.1g，2.93mmol)溶于四氢呋喃(62mL)中，加入Pd/C(0.62g，10wt.％)，氢气氛下，室温搅拌22小时。过滤浓缩得2.7g产物1-S01，收率95.1％。
MS-ESI：m/z 988.2[M+Na] +。
第十步
氮气氛下，将化合物1-S01(492mg，509.6μmol)与PyBOP(510mg，981μmol)溶于无水DMF(8.4mL)中，滴加化合物Dendrimer 1(600mg，12.74μmol)与DIPEA(127mg，981μmol)的无水DMF(8.4mL)混合溶液，反应2小时。将反应液用乙腈(16.8mL)稀释后，通过超滤(10KD，
Figure PCTCN2021089174-appb-000063
)装置，用乙腈超滤纯化，得到0.88g粗品。用纯化水(100mL)溶解，经滤膜(0.22μm)过滤，冻干后得到0.85g产物1-S00，收率103％。
1H NMR指示22DTX/树枝状聚合物。实际的分子量大约为64.3kDa(27.6％重量％DTX)。
1H NMR(400MHz，CD 3OD)δ7.99-8.25(m,55H),7.05-7.86(m,253H),6.00-6.31(m,22H),5.57-5.74(m,22H),5.11-5.48(m,76H),4.96-5.08(m,40H),4.10-4.68(m,125H),3.40-4.08(m,3200H),3.33-3.37(s,104H),3.07-3.24(m,116H),2.14-2.69(m,205H),1.00-2.10(m,1258H)。
实施例6：本公开化合物1-T00的制备
Figure PCTCN2021089174-appb-000064
第一步
将化合物1-T01-3B(42.83g，108.57mmol，根据文献European Journal of Organic Chemistry,2009,22,p.3795–3800合成)和DMAP(1.33g，10.86mmol)溶于二氯 甲烷(200mL)中，冷却至0-5℃，分批加入DCC(24.64g，119.43mmol)，升至室温搅拌21小时。减压浓缩后经柱层析色谱法纯化得34.7g产品1-T01-4，收率71％。
MS-ESI：m/z 451.2[M+H] +。
第二步
将LDA(42mL，2M在THF/己烷中)用四氢呋喃(300mL)稀释，冷却至-70℃，滴加化合物1-T01-4(17.35g，38.50mmol)的四氢呋喃(40mL)溶液，保持-70℃搅拌1小时。滴加1,3,2-二氧杂噻吩烷-2,2-二氧化物(7.17g，57.76mmol)的四氢呋喃(10mL)溶液，随后滴加DMPU(5.43g，57.76mmol)的四氢呋喃(30mL)溶液。滴毕，缓慢升至室温，搅拌16小时。加入冰水(400mL)淬灭，用二氯甲烷萃取，减压浓缩后经柱层析色谱法纯化，得4.4g产品1-T01-5，收率37.4％。
MS-ESI：m/z 477.2[M+H] +。
第三步
将化合物1-T01-5(8.76g，18.38mmol)溶于二氯甲烷(180mL)中，加入三乙基硅烷(2.56g,22mmol)，冷却至0-5℃，加入三氟乙酸(0.9mL)的二氯甲烷(2mL)溶液，升至室温搅拌19小时。浓缩后通过柱层析色谱法纯化，得1.68g产品1-T01-6，收率52％。
第四步
将化合物1-T01-6(480mg，2.75mmol)和K 2CO 3(760mg，5.51mmol)加入到DMF(5mL)中，滴加溴乙酸苄酯(758mg，3.31mmol)。室温搅拌16小时。加冰水淬灭，用二氯甲烷萃取，浓缩后经柱层析色谱法纯化，得620mg产品1-T01-7，收率69.8％。
MS-ESI：m/z 345.2[M+Na] +。
第五步
将化合物1-T01-7(620mg，1.92mmol)溶于二氯甲烷(5.0mL)中，冷却至0-5℃，滴加三氟乙酸(2.5mL)。升至室温搅拌16小时。浓缩后经柱层析色谱法纯化，得396mg产品1-T01-P，收率77.3％。
1H NMR(400MHz,CDCl 3)δ7.33-7.36(m,5H),5.16(s,2H),3.51(s,2H),1.41-1.48(m,2H),1.12-1.16(m,2H)。
第六步
氮气氛下，将化合物多西他赛(80mg，0.1mmol)和1-T01-P(53mg，0.2mmol)溶于二氯甲烷(3mL)中，加入DMAP(14mg，0.12mmol)和DCC(25mg，0.12mmol)，室温搅拌过夜。浓缩后经柱层析色谱法纯化，得70mg产品1-T01-Bn，收率67％。
MS-ESI：m/z 1057.2[M+H] +。
第七步
将化合物1-T01-Bn(70mg，0.067mol)溶于四氢呋喃(10mL)中，加入Pd/C(30mg，10wt.％)，氢气氛下，室温搅拌22小时。过滤后浓缩得60mg产物1-T01，收率：94％。
MS-ESI：m/z 988.2[M+Na] +。
第八步
氮气氛下，将化合物1-T01(59.8mg，62.0μmol)与PyBOP(62.5mg，120.0μmol)溶于无水DMF(2mL)中，滴加化合物Dendrimer 1(70mg，1.55μmol)与DIPEA(15.5mg，120.0μmol)的无水DMF(2mL)混合溶液，反应1.5小时。将反应液直接通过柱层析色谱法纯化，得到74mg粗品。用纯化水(20mL)溶解，经滤膜(0.22μm)过滤，冻干后得到71mg产物1-T00，收率73％。
1H NMR指示23DTX/树枝状聚合物。实际的分子量大约为63.2kDa(29.4％重量％DTX)。
1H NMR(400MHz,CD 3OD)δ8.02-8.25(m,54H),7.14-7.88(m,223H),6.00-6.31(m,23H),5.54-5.72(m,26H),5.14-5.51(m,94H),4.96-5.08(m,46H),4.06-4.53(m,106H),3.40-4.04(m,3100H),3.33-3.36(s,87H),3.03-3.24(m,114H),2.24-2.62(m,167H),1.00-2.10(m,1191H)。
实施例7：本公开化合物1-Z00-A的制备
Figure PCTCN2021089174-appb-000065
Figure PCTCN2021089174-appb-000066
第一步
氮气氛下，将化合物1-Z01-A-6(220mg，1.3mmol，根据文献Molecules,2020,25,2153合成)溶于四氢呋喃(5mL)中，冷却至0℃，分批加入NaH(104mg，2.2mmol，60％在油中)，控制内温<20℃，搅拌15分钟。滴加化合物S2(360mg，1.6mmol)的四氢呋喃溶液，室温搅拌过夜。将反应液冷至0℃，用饱和NaHCO 3水溶液(10mL)淬灭，用乙酸乙酯(10mL×2)萃取，浓缩后经柱层析色谱法纯化得180mg产品1-Z01-A-7，收率43％。
MS-ESI：m/z 342.2[M+Na] +。
第二步
氮气氛下，将化合物1-Z01-A-7(180mg，0.56mmol)溶于二氯甲烷(2mL)中，冷却至0℃，加入三氟乙酸(2mL)。升至室温搅拌过夜。浓缩后经柱层析色谱法纯化，得50mg产品1-Z01-A-P，收率32％。
MS-ESI：m/z 264.2[M+H] +。
第三步
氮气氛下，将化合物多西他赛(120mg，0.15mmol)和1-Z01-A-P(48mg，0.18mmol)溶于DMF(1mL)中，加入DMAP(19mg，0.15mmol)和DCC(38mg，0.18mmol)，室温搅拌过夜。浓缩后经柱层析色谱法纯化，得50mg产品 1-Z01-A-Bn，收率32％。
MS-ESI：m/z 1053.3[M+H] +。
第四步
氮气氛下，将化合物1-Z01-A-Bn(50mg，0.047mmol)溶于甲醇(3mL)中，加入Pd/C(10mg，10wt.％)，氢气氛下，室温搅拌过夜。过滤，浓缩后得35mg产品1-Z01-A，收率77％。
MS-ESI：m/z 963.3[M+H] +。
第五步
氮气氛下，将化合物1-Z01-A(34mg，35.5μmol)与PyBOP(36mg，68.5μmol)溶于无水DMF(1.0mL)中，滴加化合物Dendrimer 1(40mg，0.89μmol)与DIPEA(8.8mg，68.5μmol)的无水DMF(1.0mL)混合溶液，反应2小时。将反应液直接经柱层析色谱法纯化，得到42mg粗品，用纯化水(10mL)溶解，经滤膜(0.22μm)过滤，冻干后得到39mg产物1-Z00-A，收率69％。
1H NMR指示23DTX/树枝状聚合物。实际的分子量大约为63.1kDa(29.4％重量％DTX)。
1H NMR(400MHz,CD 3OD)δ8.00-8.20(m,53H),7.13-7.85(m,218H),5.94-6.20(m,23H),5.53-5.71(m,25H),5.15-5.40(m,91H),4.95-5.07(m,42H),4.10-4.60(m,148H),3.40-4.04(m,3100H),3.33-3.37(s,102H),3.03-3.24(m,127H),2.20-2.67(m,240H),1.00-2.10(m,1350H)。
实施例8：本公开化合物1-Z00-B的制备
Figure PCTCN2021089174-appb-000067
Figure PCTCN2021089174-appb-000068
第一步
氮气氛下，将化合物1-Z01-B-4(10.3g，47.9mmol，根据专利WO2016205633A合成)溶于DMF(150mL)中，加入碳酸钾(33.1g，239.5mmol)和苄化溴(10.6g，62.3mmol)，加热至60℃，反应16小时。冷却后加水(300mL)淬灭，用乙酸乙酯萃取，浓缩经柱层析色谱法纯化得到13g化合物1-Z01-B-5，收率89％。
MS-ESI：m/z 328.2[M+Na] +。
第二步
将化合物1-Z01-B-5(9.8g，32.1mmol)溶于二氯甲烷(50mL)中，冷却至0℃，加入三氟乙酸(30.07g，269mmol)，升至室温反应6小时。浓缩后得到15.0g粗品1-Z01-B-6，收率>100％。
MS-ESI：m/z 206.1[M+H] +。
第三步
将化合物1-Z01-B-6(7.0g，15.0mmol)溶于乙腈(70mL)中，加入碳酸钾(10.3g，75.0mmol)和溴乙酸叔丁酯(3.8g，19.5mmol)，室温反应16小时。浓缩后经柱层析色谱法纯化，得到3.0g产物1-Z01-B-7，收率62％。
MS-ESI：m/z 342.2[M+Na] +。
第四步
将化合物1-Z01-B-7(2.8g，8.8mmol)溶于二氯甲烷(10mL)中，冷却至0℃， 加入三氟乙酸(15g，135mmol)，升至室温反应16小时。浓缩后经柱层析色谱法纯化，得2.2g产品1-Z01-B-P，收率95％。
1HNMR(400MHz,CDCl 3)δ7.29-7.40(m,5H),5.15(s,2H),3.82(s,2H),2.79(s,3H),1.46-1.53(m,2H),1.32-1.40(m,2H)。
MS-ESI：m/z 264.2[M+H] +。
第五步
氮气氛下，将化合物1-Z01-B-P(1.1g，4.16mmol)，多西他赛(2.8g，3.47mmol)和DMAP(0.508g，4.16mmol)溶于无水DMF(30mL)中，冷却至0℃，加入EDCI(0.798g，4.16mmol)，升至室温反应16小时。加水(50mL)淬灭，过滤得粗品，经柱层析色谱法纯化得2.7g产品1-Z01-B-Bn，收率74％。
MS-ESI：m/z 1053.3[M+H] +。
第六步
将化合物1-Z01-B-Bn(2.7g，2.56mmol)溶于无水四氢呋喃(30mL)中，加入Pd/C(270mg，10wt.％)，氢气氛下，于室温反应16小时。过滤，浓缩后得2.3g产品1-Z01-B，收率93％。
1HNMR(400MHz,CDCl 3)δ8.13-8.10(m,2H),7.63-7.59(m,1H),7.53-7.48(m,2H),7.42-7.37(m,3H),7.30-7.26(m,3H),6.24-6.23(bs,1H),5.70-5.67(m,1H),5.49-5.43(m,2H),5.22-5.17(m,2H),4.33-4.11(m,4H)，3.93-3.91(m,1H)，3.68-3.53(m,2H),2.60-2.10(m,5H),1.94-1.68(m,10H),1.40-1.24(m,16H),1.18-1.05(m,6H)。
MS-ESI：m/z 963.3[M+H] +。
第七步
氮气氛下，将化合物1-Z01-B(573mg，594.8μmol)与PyBOP(596mg，1145μmol)溶于无水DMF(9.8mL)中，滴加化合物Dendrimer 1(700mg，14.87μmol)与DIPEA(148mg，1145μmol)的无水DMF(9.8mL)混合溶液，反应2小时。将反应液用乙腈(19.6mL)稀释后，通过超滤(10KD，
Figure PCTCN2021089174-appb-000069
)装置，用乙腈超滤纯化，得到1.01g粗品，用纯化水(100mL)溶解，经滤膜(0.22μm)过滤，冻干后得到0.99g产物1-Z00-B，收率100％。
1H NMR指示22DTX/树枝状聚合物。实际的分子量大约为64.2kDa(27.6％重量％DTX)。
1HNMR(400MHz,CD 3OD)δ8.01-8.23(m,57H),7.10-7.89(m,264H),6.00-6.31(m,22H),5.54-5.73(m,24H),5.20-5.51(m,82H),4.96-5.08(m,29H),4.10-4.68(m,132H),3.40-4.04(m,3200H),3.33-3.37(s,100H),3.07-3.24(m,97H),2.14-2.66(m,259H),0.94-2.08(m,1208H)。
实施例9：本公开化合物2-B00的制备
Figure PCTCN2021089174-appb-000070
第一步
氮气氛下，将化合物1-Z01-B4-4(6.7g，36.0mmol，根据文献Molecules,2020,25,2153合成)和无水碳酸钾(10.0g，72.4mmol)加入到无水乙腈(67mL)中，加入溴乙酸苄酯(9.9g，43.2mmol)，室温反应16小时。过滤后浓缩，粗品经柱层析色谱法纯化得10.1g 1-Z01-B4-5-Bn，收率88％。
MS-ESI：m/z 320.2[M+H] +。
第二步
将化合物1-Z01-B4-5(10.1g，31.6mmol)溶于四氢呋喃(100mL)中，加入Pd/C(1.0g，10wt.％)，氢气氛下，室温反应16小时。过滤后浓缩，粗品经柱层析色谱法纯化得5.5g 1-Z01-B4，收率76％。
1HNMR(400MHz,CDCl 3)δ3.58(s,2H),2.57(s,3H),1.45(s,9H),1.32-1.40(m,2H),0.98-1.03(m,2H)。
MS-ESI：m/z 230.1[M+H] +。
第三步
氮气氛下，将化合物2(4.25g,4.5mmol，根据专利WO2012017251合成)溶于二氯甲烷(70mL)中，加入DMAP(165mg，1.35mmol)和化合物1-Z01-B4(1.85g，8.1mmol)，冷却至0℃，滴加DCC(1.67g，8.1mmol)的DCM(10mL)溶液，升至室温搅拌过夜。过滤后减压浓缩，粗品经柱层析色谱法分离，得4.2g产物2-B00-1，收率80％。
MS-ESI：m/z 1156.3[M+H] +。
第四步
氮气氛下，将化合物2-B00-1(3.0g，2.6mmol)溶于乙腈(21mL)中，加入2,6-二甲基吡啶(4.18g，39mmol)，冷却至5℃，缓慢加入三氟甲磺酸三甲基硅酯(TMSOTf)(8.67g，39mmol)，升至室温搅拌2小时。浓缩后加乙酸乙酯(60mL)和水(40mL)，分离有机相。加入1.0M的四丁基氟化铵(TBAF)的四氢呋喃溶液(26mL，26mmol)，室温搅拌2小时。浓缩后经柱层析色谱法纯化，得2.6g产物2-B00-2，收率92％。
MS-ESI：m/z 1100.2[M+H] +。
第五步
氮气氛下，将化合物2-B00-2(1.38g，1.25mmol)与PyBOP(0.79g，1.51mmol)溶于无水DMF(21mL)中，滴加化合物Dendrimer 1-PEG2K(2.00g，0.025mmol)与N-甲基吗啉(NMM)(0.51g，5.03mmol)的无水DMF(21mL)混合溶液，室温反应2小时。将反应液用乙腈(42mL)稀释后，通过超滤(10KD，
Figure PCTCN2021089174-appb-000071
)装置，用乙腈超滤纯化，得到2.25g粗品，用纯化水(150mL)溶解，经滤膜(0.22μm)过滤，冻干后得到2.20g产物2-B00，收率89％。
1H NMR指示21个药物分子/树枝状聚合物。实际的分子量大约为98.5kDa(20.1％重量％药物分子)。
1H NMR(400MHz,CD 3OD)δ6.40-8.52(m,420H),4.13-4.70(m,62H),3.40-4.10(m,6000H),3.33-3.36(s,127H),2.79-3.25(m,279H),2.22-2.74(m,225H),0.61-1.94(m,606H).
生物学评价
测试例1：本公开化合物在人和小鼠血浆中的稳定性研究
1、供试品
实施例中所得的化合物1-A00、1-B00、1-C00、1-F00、1-L00、1-M00、1-S00以及多西他赛。
2、试验用血浆
类型 品系 性别 来源
人 - 混合 Pharmaron
小鼠 CD-1 混合 Pharmaron
3、试验方法
1)溶液配制
在PBS中制备10mg/mL的测试化合物和多西他赛溶液。在乙腈中制备了1mM的丙烷茶碱工作溶液，在该试验中将丙烷茶碱用作人和小鼠血浆的阳性对照。
2)血浆稳定性的测试
a.在37℃水浴中快速解冻冷冻的血浆。在室温以3220g离心血浆10分钟，以去除凝块，并将上清液收集到新的试管中。检查并记录血浆的pH值。
b.将每个细胞的490μL血浆添加到培养板中，将培养板在37℃预热15分钟。
c.预孵育后，将10μL的10mg/mL测试化合物添加到490μL血浆中，以达到0.2mg/mL的最终浓度；将2μL的1mM对照化合物添加到398μL血浆中，以达到最终浓度为5μM。该测定将一式两份进行。
d.将反应样品在37℃孵育。
e.在0、0.5、2、6和24小时从反应样品中取出50μL的等分试样。加入350μL含内标的冷乙腈终止反应。
f.将所有样品涡旋10分钟，然后在3220g离心30分钟以沉淀蛋白质。将100μ0上清液转移至新板中。上清液将根据LC-MS信号响应和峰形用超纯水稀释。
3)样品分析
样品通过LC-MS/MS进行分析
4)数据分析
所有计算均使用Microsoft Excel进行。从提取的离子色谱图中确定峰面积比。通过以下公式计算每个时间点的转化浓度：
转化浓度 t小时(μg/mL)＝峰面积比 t小时/峰面积比 STD×200(μg/mL)
其中峰面积比 t小时是t时测试化合物的峰面积比；
峰面积比 STD是200μg/mL多西他赛的峰面积比。
4、试验结果
本公开化合物在人和小鼠血浆中的转化数据如下表1和表2以及图1所示。
表1人血浆中的转化数据
Figure PCTCN2021089174-appb-000072
Figure PCTCN2021089174-appb-000073
表2小鼠血浆中的转化数据
Figure PCTCN2021089174-appb-000074
注：△c指的是最近两个时间点的血浆转化浓度差值
从表1和表2及图1可以看出，本公开化合物具有平稳释放药物的效果。其中化合物1-S00的突释更低，药物释放更好，且在达到治疗浓度的基础上，24小时内释放更加平稳(从表中的△c值可对比得出)，通过药物缓释修饰体内药代动力学特征，可以降低副作用，提高治疗窗。
测试例2：单次静脉注射给予比格犬不同化合物的药代动力学研究
1、供试品
实施例中所得的化合物2-B00以及2-A00(根据专利WO2018154004合成，连接Dendrimer 1-PEG2K)。
样品制备方法：120mg/瓶的受试物，加入终体积5％DMSO、95％生理盐水，涡旋溶解得到1.5mg/ml的供试样品。
2、试验动物
比格犬，
Figure PCTCN2021089174-appb-000075
级别，来源为美迪西普亚储备动物库：999M-004。
3、试验方法
将收集的血液样本置于EDTA-K2抗凝型采血管中,血液样本均需加酯酶抑制剂进行特殊处理。在EDTA-K2抗凝的采血管中加入1/40采血体积的200mM DDVP(如采血量为0.04mL，则加入1μL 200mM DDVP)。将收集到的全血放在冰上，1小时内离心分离血浆(离心力2200g，离心10min，2-8℃)。测定血药浓度。
表3给药方案
Figure PCTCN2021089174-appb-000076
Figure PCTCN2021089174-appb-000077
4、试验结果
各组的临床观察情况及药代动力学数据表4及图2所示。
表4药代动力学数据
Figure PCTCN2021089174-appb-000078
从试验结果可以看出，本公开化合物的早期药物释放更慢，在达到治疗浓度的基础上，释放行为更加平稳。其中2-B00相比于2-A00，在AUC近似的情况下，拥有更低的C max，可以降低副作用，提高治疗窗。

Claims (23)

  1. 一种大分子，其包含：
    i)具有表面氨基的树枝状聚合物D，其中至少两个不同的端基共价连接于所述树枝状聚合物的表面氨基：
    ii)第一端基，其为包含羟基、氨基、羧基或者巯基的药学活性剂或其残基A；
    iii)第二端基，其为药代动力学修饰剂；
    其中所述第一端基通过连接子-X 1-L-X 2-共价连接于所述树枝状聚合物的表面氨基，X 1为连接子与药学活性剂的连接基，X 2为连接子与树枝状聚合物的连接基，其中：
    a)X 1和X 2选自-C(O)-或不存在，L为任选被一个或多个氧、硫或氮原子间断的C 1-10直链或支链亚烷基；当X 1和X 2中至少一个不存在时，所述C 1-10直链或支链亚烷基任选被取代基取代；当X 1和X 2均为-C(O)-时，所述C 1-10直链或支链亚烷基进一步被取代基取代；所述的取代基选自氘、羟基、C 3-7环烷基、C 3-7亚环烷基、烷氧基、卤代烷基、卤代烷氧基、卤素、硝基、氰基、酰基、硫醚基、亚磺酰基、磺酰基、-NR 1R 2、芳基、杂芳基和杂环基一个或多个基团；或者
    b)X 1和X 2选自-C(O)-或不存在，L为任选被一个或多个氧、硫或氮原子间断的C 1-10直链或支链亚烯基或亚炔基，所述亚烯基和亚炔基任选被选自氘、羟基、烷基、环烷基、烷氧基、卤代烷基、卤代烷氧基、卤素、硝基、氰基、酰基、硫醚基、亚磺酰基、磺酰基、-NR 1R 2、芳基、杂芳基和杂环基的一个或多个基团所取代；或者
    c)X 1选自-C(O)-或不存在，X 2不存在，L与树枝状聚合物表面氨基的氮原子连接后形成含氮杂环结构
    Figure PCTCN2021089174-appb-100001
    L 1为任选被一个或多个氧、硫或氮原子间断的C 1-10直链或支链亚烷基、亚烯基或亚炔基，所述亚烷基、亚烯基或亚炔基任选被选自氘、羟基、烷基、环烷基、亚环烷基、烷氧基、卤代烷基、卤代烷氧基、卤素、硝基、氰基、酰基、硫醚基、亚磺酰基、磺酰基、-NR 1R 2、芳基、杂芳基和杂环基的一个或多个基团所取代，X 3选自CR 5或N，其中R 5选自氢、氘、羟基、烷基或烷氧基，m选自0、1或2；或者
    d)X 1选自-C(O)-或不存在，X 2不存在，且L为
    Figure PCTCN2021089174-appb-100002
    L 2为任选被一个或多个氧、硫或氮原子间断的C 1-10直链或支链亚烷基、亚烯基或亚炔基，所述亚烷基、亚烯基或亚炔基任选被选自氘、羟基、烷基、环烷基、烷氧基、卤代烷 基、卤代烷氧基、卤素、硝基、氰基、酰基、硫醚基、亚磺酰基、磺酰基、-NR 1R 2、芳基、杂芳基和杂环基的一个或多个基团所取代，n选自1或2，其中含氮杂环端与树枝状聚合物连接，L 2端通过-C(O)-或者直接与药学活性剂连接；
    其中，R 1、R 2各自独立地选自氢、羟基、C 1-6烷基、环烷基、C 1-6烷氧基。
  2. 根据权利要求1所述的大分子，其中X 1和X 2均为-C(O)-，L为被一个或多个氧、硫或氮原子间断的C 1-6直链或支链亚烷基，所述C 1-6直链或支链亚烷基被独立地选自氘、羟基、C 3-7环烷基、C 3-7亚环烷基、C 1-6烷氧基、C 1-6卤代烷基、C 1-6卤代烷氧基和卤素的一个或多个取代基所取代。
  3. 根据权利要求2所述的大分子，其中X 1和X 2均为-C(O)-，L为被一至三个独立地选自硫或氮的原子间断的C 1-6直链或支链亚烷基，所述C 1-6直链或支链亚烷基被独立地选自羟基、C 3-7环烷基、C 3-7亚环烷基、C 1-6烷氧基、C 1-6卤代烷基、C 1-6卤代烷氧基和卤素的一个或多个取代基所取代。
  4. 根据权利要求3所述的大分子，其中所述C 1-6直链或支链亚烷基被一个或多个C 3-7亚环烷基取代。
  5. 根据权利要求1所述的大分子，其中连接子为-C(O)-X-C(O)-，
    其中X为
    Figure PCTCN2021089174-appb-100003
    R a、R b各自独立地选自氢、C 1-6烷基，或R a、R b与其相连的碳原子一起形成C 3-7环烷基；R c、R d各自独立地选自氢、C 1-6烷基，或R c、R d与其相连的碳原子一起形成C 3-7环烷基；条件是至少有一个R a、R b与其相连的碳原子一起形成C 3-7环烷基或至少有一个R c、R d与其相连的碳原子一起形成C 3-7环烷基；
    X’选自-CH 2-、-O-、-S-和-NR 3-；
    R 3为氢或C 1-6烷基；
    s和t各自独立地选自1至4的整数。
  6. 根据权利要求5所述的大分子，其中X为
    Figure PCTCN2021089174-appb-100004
    Figure PCTCN2021089174-appb-100005
    其中s1和t1各自独立地选自1至4的整数。
  7. 根据权利要求5或6所述的大分子，其中X’选自-O-、-S-、-NH-和-N(CH 3)-。
  8. 根据权利要求1所述的大分子，其中连接子-X 1-L-X 2-中，X 1为-C(O)-，与药学活性剂或其残基A连接；X 2为-C(O)-，与树枝状聚合物D的表面氨基连接形成酰胺键，结构如下：
    Figure PCTCN2021089174-appb-100006
  9. 根据权利要求1所述的大分子，其中连接子-X 1-L-X 2-中，X 1为-C(O)-，与药学活性剂或其残基A连接；X 2为-C(O)-，与树枝状聚合物D的表面氨基连接形成酰胺键，结构如下：
    Figure PCTCN2021089174-appb-100007
  10. 根据权利要求1所述的大分子，其中连接子-X 1-L-X 2-中，X 1为-C(O)-，与药学活性剂或其残基A连接；X 2不存在，L的结构如下，其中含氮杂环端与树枝状聚合物D的表面氨基连接形成碳-氮键：
    Figure PCTCN2021089174-appb-100008
  11. 一种大分子，其包含：
    i)具有表面氨基的树枝状聚合物D，其中至少两个不同的端基共价连接于所述树枝状聚合物的表面氨基：
    ii)第一端基，其为包含羟基、氨基、羧基或者巯基的药学活性剂或其残基A；
    iii)第二端基，其为药代动力学修饰剂；
    其中所述第一端基通过三酸连接子共价连接于所述树枝状聚合物的表面氨基，三酸连接子中的一个羰基与药学活性剂或其残基连接，另外两个羰基与树枝状聚合物的同一表面氨基连接并因此与连接子部分形成含氮杂环。
  12. 根据权利要求11所述的大分子，其中所述的三酸连接子与药学活性剂或其残基A以及树枝状聚合物D连接，形成如下的结构：
    Figure PCTCN2021089174-appb-100009
    其中X 4选自C、N，L 3为任选被氧、硫或氮原子间断的C 1-10亚烷基、亚烯基或者亚炔基，p为选自0-5的整数。
  13. 根据权利要求11所述的大分子，其中所述的三酸连接子与药学活性剂或其残基A以及树枝状聚合物D连接，形成选自如下的结构：
    Figure PCTCN2021089174-appb-100010
  14. 一种大分子，其包含：
    i)具有表面氨基的树枝状聚合物D，其中至少两个不同的端基共价连接于所述树枝状聚合物的表面氨基：
    ii)第一端基，其为艾日布林、曲贝替定或卢比替定；
    iii)第二端基，其为药代动力学修饰剂；
    其中所述第一端基通过二酸连接子共价连接于所述树枝状聚合物的表面氨基，所述二酸连接子包含任选被一个或多个硫或氮原子间断的C 1-10烷基链。
  15. 根据权利要求1至13中任一项所述的大分子，其中药学活性剂选自麻醉剂、抗酸剂、抗体、抗感染药、生物制品、心血管药物、造影剂、利尿剂、补血药、免疫抑制剂、激素和类似物、营养制品、眼科药物、疼痛治疗剂、呼吸药物、 佐剂、同化剂、抗关节炎药、抗惊厥药、抗组胺药、消炎药、抗溃疡药、行为矫正药物、肿瘤药物、中枢神经系统药物、避孕药、糖尿病治疗药物、生育药物、生长促进剂、止血药、免疫刺激剂、肌肉松弛药、肥胖治疗剂、骨质疏松症药物、肽、镇静剂和安定药、尿道酸化剂或维生素。
  16. 根据权利要求1-13任一项所述的大分子，其中所述的药学活性剂选自紫杉烷类药物、喜树碱衍生物、核苷类药物、蒽环类药物、海鞘素衍生物、蛋白酶体抑制剂、微管抑制剂、BCL-2抑制剂、BCL-X L抑制剂、选择性核输出抑制剂、抗代谢药物、酪氨酸激酶抑制剂、PLK1抑制剂、CDK4/6抑制剂、BTK抑制剂、非甾体激素受体拮抗剂和类固醇，优选紫杉烷类药物、BCL-2抑制剂和BCL-X L抑制剂。
  17. 根据权利要求1至13中任一项所述的大分子，其中药学活性剂选自多西他赛、伊立替康、吉西他滨、卡培他滨、地西他滨、阿扎胞苷、多柔比星、表柔比星、曲贝替定、卢比替定、硼替佐米、艾日布林、塞利尼索、维奈托克、特西他赛、培美曲塞、卡巴他赛、卡博替尼、昂凡色替，或这些药物分子的结构改造物。
  18. 根据权利要求1至13中任一项所述的大分子，其中药学活性剂为多西他赛或下式化合物2，或它们的结构改造物，
    Figure PCTCN2021089174-appb-100011
  19. 根据权利要求1至18中任一项所述的大分子，其中所述的药代动力学修饰剂选自聚乙二醇、聚乙基噁唑啉、聚乙烯基吡咯烷酮、聚丙二醇、叶酸盐或者关于细胞表面受体的配体的叶酸盐衍生物，优选聚乙二醇。
  20. 根据权利要求1至18中任一项所述的大分子，其中所述树枝状聚合物选 自聚赖氨酸、聚赖氨酸类似物、聚酰氨基胺(PAMAM)、聚乙烯亚胺(PEI)或聚醚羟胺(PEHAM)树枝状聚合物，优选聚赖氨酸或者聚赖氨酸类似物。
  21. 根据权利要求20所述的大分子，其中所述聚赖氨酸或聚赖氨酸类似物包含核心和2-7代赖氨酸或赖氨酸类似物。
  22. 一种药物组合物，包含根据权利要求1至21中任一项所述的大分子和药学上可接受的载体。
  23. 根据权利要求14、16-18中任一项所述的大分子在制备治疗肿瘤的药物中的用途。
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CN104650194A (zh) * 2015-02-14 2015-05-27 四川大学 一种肽类树状大分子药物及其制备方法和应用
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