WO2014086127A1 - Doxorubicin conjugate medicine and preparation method thereof - Google Patents

Doxorubicin conjugate medicine and preparation method thereof Download PDF

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Publication number
WO2014086127A1
WO2014086127A1 PCT/CN2013/073164 CN2013073164W WO2014086127A1 WO 2014086127 A1 WO2014086127 A1 WO 2014086127A1 CN 2013073164 W CN2013073164 W CN 2013073164W WO 2014086127 A1 WO2014086127 A1 WO 2014086127A1
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doxorubicin
formula
repeating unit
carboxylated
acid
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PCT/CN2013/073164
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French (fr)
Chinese (zh)
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庄秀丽
丁建勋
张建成
陈学思
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中国科学院长春应用化学研究所
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Publication of WO2014086127A1 publication Critical patent/WO2014086127A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/02—Acyclic radicals, not substituted by cyclic structures
    • C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
    • A61K47/551—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia

Definitions

  • the invention relates to the technical field of polymer drugs, in particular to doxorubicin bonding drugs and a preparation method thereof.
  • Doxorubicin also known as hydroxydaunorubicin, hydroxyd-dentamicin, doxorubicin or ADR, is a broad-spectrum anti-tumor drug of anthracycline antibiotics, a cell cycle non-specific drug with the strongest effect on S phase. It also has effects on ⁇ 1, G1 and G2 phases. Its mechanism of action is to embed DNA molecules and destroy nucleic acid structures to inhibit the synthesis of biological macromolecules.
  • Doxorubicin is mainly used for the treatment of acute leukemia, malignant lymphoma, liver cancer, lung cancer, gastric cancer, esophageal cancer, cervical cancer, bladder cancer, testicular cancer, verrucous adenocarcinoma, soft tissue tumor, osteosarcoma and neuroblastoma.
  • the main clinical mode of administration of doxorubicin is intravenous infusion.
  • the drug is rapidly distributed throughout the body, thus lacking the target for tumor tissue, and the ratio of doxorubicin that can reach the tumor site and exert its therapeutic effect is very high. Low, resulting in low bioavailability and inefficiency. Therefore, clinical administration requires frequent administration, thereby causing a large toxic side effect on normal tissues and organs of the body.
  • doxorubicin In order to solve the clinical problems of doxorubicin, the development of new doxorubicin dosage forms has been a hot spot for researchers. Among them, nanoscale drug delivery systems have been developed rapidly, including liposomes, polymer nanoparticles, and micelles. Among them, micelles prepared from amphiphilic block copolymers or graft copolymers have become the focus of research on drug carriers. There are two main ways in which micelles are loaded with drugs, one is physical encapsulation and the other is chemical bonding. Among them, chemical bonding has been more widely studied due to the more stable drug entrapment, the more sustained drug release process, and longer drug release times.
  • the Chinese Patent Publication No. CN101234204 A discloses a polymer doxorubicin-bonding compound, which performs a ring-opening polymerization reaction of an aliphatic cyclic ester under the action of a solvent of a polyethylene glycol and a catalyst to obtain a polyethylene glycol- Polylactic acid block copolymer; the terminal hydroxyl group of the block copolymer is converted into a terminal carboxyl group, and amidation reaction with doxorubicin under the action of a condensing agent to obtain a polymer
  • CN101234205 A discloses a polymer-type doxorubicin-bonded drug having a targeting function, which is assembled by mixing two kinds of polyethylene glycol-polylactic acid block copolymers.
  • the polylactic acid chain block copolymer of the first type of polyethylene glycol-polylactic acid block copolymer is linked with doxorubicin, and the polyethylene glycol chain end of the second polyethylene glycol-polylactic acid block copolymer is linked with lactose.
  • Doxorubicin is protected by both polyethylene glycol and polylactic acid, and has a sustained release function; lactose has a targeting function and can achieve targeted delivery of doxorubicin.
  • Biomaterials discloses a carboxyl group modified with polyethylene glycol monoterpene ether and cis-3-carboxypentadienoic anhydride on the surface amino group of a polyamide-amine dendrimer.
  • the doxorubicin thereby obtaining a polymer doxorubicin bonding drug, which can be quickly translated and released in tumor tissues and cells under acidic conditions, thereby realizing the intelligence of drug release, but the mold
  • the carrier material used for the drug bonding is a polyamide-amine dendrimer, which is cumbersome to prepare and has poor biocompatibility, which is not conducive to the practical application of the doxorubicin bond.
  • the technical problem to be solved by the present invention is to provide an adriamycin bonding drug and a preparation method thereof, and the doxorubicin bonding drug provided by the invention has simple preparation process, good biocompatibility, and can be realized in a tumor site. Aggregation with pH responsiveness and active targeting.
  • the present invention provides an adriamycin-bonding drug comprising a first repeating unit having the structure of the formula (I), a second repeating unit of the formula (II), a third repeating unit of the formula (III), and the formula (IV)
  • R is selected from one of the structures of the formula (1) to (4):
  • Lactonibic acid is lactobionic acid, which has the structure of formula (5):
  • FA is folic acid, which has the structure of formula (6):
  • the number of moles of the first repeating unit, the number of moles of the second repeating unit, the number of moles of the third repeating unit, the number of moles of the fourth repeating unit, and the first repeating unit and the second repeating unit satisfies the following condition: 0.02 ⁇ h / (h + i + j + k) ⁇ 0.3; 0 ⁇ i / (h + i + j + k) ⁇ 0.3; 0 ⁇ j / (h + i + j + k) ⁇ 0.3.
  • the doxorubicin bond has a number average molecular weight of 6000 to 100,000.
  • the invention also provides a preparation method of an adriamycin bonding drug, comprising the following steps:
  • the molar ratio of the carboxylated doxorubicin derivative, lactobionic acid, folic acid to the glucose repeating unit on the dextran backbone is (2 ⁇ 30): (0 ⁇ 30): (0 ⁇ 30): 100 .
  • the molar ratio of the carboxylated doxorubicin derivative, lactobionic acid, folic acid to the glucose repeating unit on the dextran backbone is (8 ⁇ 25): (5 ⁇ 25): (5 ⁇ 25): 100 .
  • the dextran has a molecular weight of 6000 to 100,000.
  • the organic solvent is dimethyl sulfoxide.
  • the carboxylated doxorubicin derivative is prepared according to the following method:
  • Doxorubicin hydrochloride, anhydride and triethylamine are reacted in an organic solvent to obtain a carboxylated doxorubicin derivative;
  • the anhydride is selected from the group consisting of 1,2-dicarboxycyclohexene anhydride, succinic anhydride, 2 One of 2,3,3-tetradecylsuccinic anhydride and cis-3-carboxypentenedioic anhydride.
  • the molar ratio of the doxorubicin hydrochloride, anhydride and triethylamine is 1: (0.5 ⁇ 2): (0.5-2).
  • the present invention is derived from glucan, carboxylated doxorubicin having the structure of formula (II-a), formula (II-b), formula (II-c) or formula ( ⁇ -d).
  • the substance, lactobionic acid and folic acid are used as raw materials to cause a condensation reaction to obtain an adriamycin bonding drug.
  • lactobionic acid and folic acid have active targeting functions, and the doxorubicin bonding agent provided by the invention can achieve drug accumulation at the tumor site through enhanced penetration and retention effects and active targeting function.
  • the polymer doxorubicin-bonded drug provided by the invention has pH responsiveness, and the doxorubicin bonded to the polymer drug by the amide bond can be quickly translated under the pH condition of the tumor tissue or the tumor cell. Thereby enhancing the effect of the drug.
  • the present invention uses glucan as a raw material, and has not only good biocompatibility, but also simple and easy to purchase, which is advantageous for the in-depth utilization of doxorubicin-based drugs.
  • Example 1 is a nuclear magnetic resonance spectrum of a carboxylated doxorubicin derivative prepared in Example 1 of the present invention
  • FIG. 2 is an infrared analysis spectrum of a carboxylated doxorubicin derivative prepared in Example 1 of the present invention
  • FIG. 3 is a nuclear magnetic resonance spectrum of a carboxylated doxorubicin derivative prepared in Example 5 of the present invention.
  • the present invention provides an adriamycin-bonding drug comprising a first repeating unit having the structure of the formula (I), a second repeating unit of the formula (II), a third repeating unit of the formula (III), and the formula (IV)
  • the fourth repeating unit of the structure is:
  • R is selected from one of the structures of the formulae (1) to (4):
  • Lactonibicacid is lactobionic acid with the structure of formula (5):
  • FA is folic acid and has the structure of formula (6):
  • the ratio of the total number of moles of the third repeating unit and the fourth repeating unit satisfies the following condition: 0.02 ⁇ h / (h + i + j + k) ⁇ 0.3; 0 ⁇ i / (h + i + j + k) ⁇ 0.3;0 ⁇ j/(h+i+j+k) ⁇ 0.3; Preferably, the following conditions are satisfied: 0.15 ⁇ h/(h+i+j+k) ⁇ 0.3, 0.10 ⁇ i/(h+i+j +k) ⁇ 0.20, 0.10 ⁇ j/(h+i+j+k) ⁇ 0.20; More preferably, the following conditions are satisfied: 0.18 ⁇ h/(h+i+j+k) ⁇ 0.28, 0.12 ⁇ i/
  • the main chain is dextran
  • the glucose repeating unit is condensed with the doxorubicin derivative to obtain a repeating unit having the structure of the formula (I);
  • the glucose repeating unit Condensation with lactobionic acid gives a repeating unit having the structure of formula (II);
  • the glucose repeating unit is condensed with folic acid to obtain a third repeating unit having the structure of formula (III).
  • the number average molecular weight of the doxorubicin bond is preferably from 6000 to 100,000, more preferably from 7,000 to 90,000, and most preferably from 8,000 to 80,000.
  • the invention also provides a preparation method of a doxorubicin bond, comprising the following steps: glucan, carboxylated doxorubicin derivative, lactobionic acid having the structure of formula (V) and folic acid having the structure of formula (VI) 1-(3-Diaminoaminopropyl)-3-ethylcarbodiimide' hydrochloride and 4-diguanidinoaminopyridine are reacted in an organic solvent to obtain an adriamycin-binding drug, the carboxylated A
  • the derivative of the formula has the structure of the formula (VII-a), the formula (VII-b), the formula (VII-c) or the formula (VII-d):
  • the invention firstly adds dextran, carboxylated doxorubicin derivative, lactobionic acid having the structure of formula (V) and folic acid having the structure of formula (VI) to an organic solvent, and then adding 1-(3-diaminoamino group).
  • the propyl)-3-ethylcarbodiimide hydrochloride is reacted with 4-diaminopyridine to give an adriamycin bond.
  • the molar ratio of the carboxylated doxorubicin derivative, lactobionic acid, folic acid to the glucose repeating unit on the dextran main chain is preferably (2 to 30): (0 to 30): (0 to 30) 100: More preferably (8 ⁇ 25): (5 ⁇ 25): (5 ⁇ 25): 100; the 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide.
  • the ratio of the number of moles of the hydrochloride to the sum of the moles of the carboxylated doxorubicin derivative, lactobionic acid and folic acid is preferably 1: (4 to 6); more preferably 1: (4.5 to 5.5);
  • the ratio of the number of moles of 4-diaminopyridine to the sum of the moles of the carboxylated doxorubicin derivative, lactobionic acid and folic acid is preferably 1: (1 to 3), more preferably 1: (1.5 ⁇ 2.5);
  • the molecular weight of the glucan is preferably from 6000 to 100000, more preferably from 7000 to 90,000, most preferably from 8,000 to 80,000;
  • the organic solvent is preferably dimercaptosulfoxide;
  • the reaction is preferably under stirring conditions Go on.
  • the reaction temperature is preferably room temperature;
  • the reaction time is preferably from 3 to 5 days, more preferably from 3.5 to 4 days.
  • the reaction solution is dialyzed into a dialysis bag, and lyophilized to obtain an adriamycin-binding drug.
  • the molecular weight cut off of the dialysis bag is preferably 3,500 Dalton; the time of the dialysis is preferably 3 to 5 More preferably, it is 3.5 to 4 days; in the dialysis process, water is preferably changed every 3 to 5 hours, more preferably 3.5 to 4.5 hours; the present invention is not limited to freeze drying, and those skilled in the art It is well known to freeze and dry.
  • the present invention has no limitation on the source of the glucan, and it is commercially available.
  • the present invention has no limitation on the source of the lactobionic acid and folic acid, and is commercially available.
  • the carboxylated doxorubicin derivative has a structure of the formula (VII-a), the formula (VII-b), the formula (VII-c) or the formula (Vll-d), preferably prepared according to the following method:
  • doxorubicin hydrochloride, the acid anhydride and the triethylamine are reacted in an organic solvent to obtain a carboxylated doxorubicin derivative, which is 1,2-dicarboxycyclohexene anhydride, succinic anhydride, 2 , 2,3,3-tetradecyl succinic anhydride or cis-3-carboxypentene dianhydride.
  • doxorubicin hydrochloride, anhydride and triethylamine are dissolved in an organic solvent, and doxorubicin hydrochloride and an acid anhydride are reacted to obtain a carboxylated doxorubicin derivative.
  • the organic solvent is preferably ruthenium, osmium-dimercaptoamide; the molar ratio of the doxorubicin hydrochloride, anhydride and triethylamine is preferably 1: (0.5 ⁇ 2): (0.5 ⁇ 2), More preferably, it is 1: (1-1.5): (1 to 1.5).
  • the doxorubicin hydrochloride and the acid anhydride are preferably reacted under stirring, and the temperature of the reaction is preferably from 10 ° C to 40 ° C, more preferably from 15 ° C to 35 ° C, and most preferably 20 °C ⁇ 30°C, the time is preferably 101! ⁇ 50h, more preferably 15h ⁇ 40h, most preferably 20h ⁇ 30h.
  • the obtained reaction mixture was poured into 20 volumes of ethyl acetate, and then washed with a saturated aqueous solution of sodium chloride, dried, filtered, and concentrated to give a carboxylated doxorubicin derivative.
  • the acid anhydride is 1,2-dicarboxycyclohexene anhydride, succinic anhydride, 2,2,3,3-tetradecyl succinic anhydride or cis-3-carboxypentene dianhydride.
  • the obtained carboxylated doxorubicin derivative has a structure of the formula (VII-a); when the acid anhydride is succinic anhydride, the obtained carbamate
  • the derivative of the formula has the structure of the formula (VII-b); when the anhydride is 2,2,3,3-tetradecylsuccinic anhydride, the obtained carboxylated doxorubicin derivative has the structure of the formula (VII-c)
  • the acid anhydride is cis-3-carboxypentadienoic anhydride
  • the obtained carboxylated doxorubicin derivative has a structure of the formula (V11-d).
  • the present invention uses dextran, a carboxylated doxorubicin derivative having the structure of formula (II-a), formula (II-b), formula (II-c ) or formula (II-d), lactobionic acid and folic acid.
  • the raw material is subjected to a condensation reaction to obtain an adriamycin-bonding drug.
  • lactobionic acid and folic acid have active targeting functions, and the present invention provides Adriamycin-based drugs can achieve drug accumulation at the tumor site through enhanced penetration and retention effects and active targeting.
  • the polymer doxorubicin-bonded drug provided by the invention has pH responsiveness, and the doxorubicin bonded to the polymer drug by the amide bond can be quickly released under the pH condition of the tumor tissue or the tumor cell. , thereby enhancing the effect of the drug.
  • the invention uses dextran as a raw material, has not only good biocompatibility, but also is easy to purchase, which is beneficial to the in-depth utilization of the doxorubicin bond.
  • FIG. 1 is a nuclear magnetic resonance spectrum of the carboxylated doxorubicin derivative prepared in Example 1 of the present invention; infrared analysis of the carboxylated doxorubicin derivative, the results are shown in FIG. 2, and FIG. 2 is an embodiment of the present invention.
  • An infrared analysis spectrum of the prepared carboxylated doxorubicin derivative, as shown in Fig. 1 and Fig. 2, the carboxylated doxorubicin derivative prepared in the examples of the present invention has a structure of the formula ( ⁇ -d).
  • Example 3 580.0 mg (0.0 billion mol) of doxorubicin hydrochloride, 126.1 mg (0.0 billion mol) of succinic anhydride and 101.2 mg (0.0 billion mol) of triethylamine were placed in a dry reaction flask, and 5 mL of anhydrous hydrazine was added. , ⁇ -dimercaptoamide is dissolved, and reacted at 25 ° C with stirring under stirring for 24 h. After the reaction is completed, the obtained reaction mixture is poured into 100 mL of ethyl acetate and diluted with a saturated aqueous solution of sodium chloride. Washing, drying, filtration, and concentration give a carboxylated doxorubicin derivative.
  • reaction mixture was subjected to a molecular weight cutoff of 3,500 Dalton.
  • the dialysis bag was dialyzed for 3 days, the water was changed every 5 hours, and the reaction solution was lyophilized to obtain an adriamycin-binding drug.
  • Example 3 is a nuclear magnetic resonance spectrum of the doxorubicin-bonded drug prepared in Example 5 of the present invention.
  • the results are as follows: 8.57 ppm, 7,95 ppm, 7.61 ppm are characteristic peaks of -FA (folic acid); 4.0 to 5.0 ppm, 3.8 to 3.0 ppm are characteristic peaks of dextran; 3.3 ppm is characteristic peak of water; 4.0 ppm is ⁇ Characteristic peak of doxorubicin derivative; 2.5ppm is a characteristic peak of deuterated DMSO.
  • the result shows that this The doxorubicin-bonded drug prepared in Inventive Example 5 has the structure of the formula (I), wherein the values of h, j, and k are shown in Table 1.
  • the polymer doxorubicin bonding drugs prepared in the preparation of 20 to 22 have values of h, j, and k.
  • the dialysis bag with a molecular weight cut off of 3500 Dalton was dialyzed for 3 days, and the water was changed every 5 hours.
  • the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
  • the nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 23 to 25 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, j and k are shown in Table 7. .
  • reaction mixture was dialyzed against a dialysis bag with a molecular weight cutoff of 3,500 Dalton for 3 days, and the water was changed every 5 hours.
  • the reaction solution was lyophilized to obtain a polymer doxorubicin bond. .
  • the polymer doxorubicin bonding drugs prepared by the preparation of the compounds 36 to 38 have values of h, i, k
  • lactobionic acid 0.29735g, 0.49560g and 0.79295g of 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide hydrochloride, 0.01895g, 0.03158g and 0.05053g of 4-diaminopyridine, after mixing uniformly, add 25mL of dimercaptosulfoxide to dissolve the reaction system, react at room temperature for 3 days under light agitation, and after the reaction is finished, separately mix the obtained reaction.
  • the solution was dialyzed against a dialysis bag having a molecular weight cutoff of 3,500 Dalton for 3 days, and the water was changed every 5 hours, and the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
  • reaction mixture is respectively subjected to molecular weight cut off.
  • the 3500 Dalton dialysis bag was dialyzed for 3 days, the water was changed every 5 hours, and the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
  • the obtained reaction mixture was dialyzed against a dialysis bag having a molecular weight cutoff of 3,500 Dalton for 3 days, and the water was changed every 5 hours. Once, the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
  • reaction mixture was dialyzed against a dialysis bag having a molecular weight cutoff of 3,500 Dalton for 3 days, and the water was changed every 5 hours.
  • the reaction solution was lyophilized to obtain a polymer doxorubicin bond.

Abstract

Provided in the present invention is a Doxorubicin conjugate medicine comprising a first repeat unit of a structure of formula (I), a second repeat unit of a structure of formula (II), a third repeat unit of a structure of formula (III) and a fourth repeat unit of a structure of formula (IV). Also provides in the present invention is a method for preparing Doxorubicin conjugate medicine. In the present invention, the Doxorubicin conjugate medicine is obtained by the condensation reaction of glucan, carboxylated doxorubicin derivative, lactose acid and folic acid as raw materials. Among them, lactose acid and folic acid have an active targeting function, and the Doxorubicin conjugate medicine provided in the present invention can realize the concentration of drugs at the tumour site through enhanced permeability and retention effects and the active targeting function. In addition, the polymer- Doxorubicin conjugate medicine provided in the present invention has pH sensitivity, wherein the doxorubicin bond on the polymer medicine via an amide bond can be quickly released under the condition of the pH within a tumour tissue or tumour cell, so as to enhance the effect of the medicine.

Description

阿霉素键合药及其制备方法 本申请要求于 2012 年 12 月 4 日提交中国专利局、 申请号为 201210512758.4、发明名称为"阿霉素键合药及其制备方法"的中国专利申 请的优先权， 其全部内容通过引用结合在本申请中。 技术领域  Doxorubicin-bonded medicine and preparation method thereof The present application claims to be Chinese patent application filed on December 4, 2012, the Chinese Patent Office, the application number is 201210512758.4, and the invention name is "adriamycin-bonding drug and its preparation method" Priority is hereby incorporated by reference in its entirety. Technical field
本发明涉及高分子药物技术领域，尤其涉及阿霉素键合药及其制备方 法。  The invention relates to the technical field of polymer drugs, in particular to doxorubicin bonding drugs and a preparation method thereof.
背景技术 Background technique
阿霉素， 又名羟柔红霉素、 羟正定霉素、 多柔比星或 ADR, 是一种 蒽环抗生素类广谱抗肿瘤药物， 是细胞周期非特异性药， 对 S 期作用最 强， 对^1、 G1和 G2期也有作用， 其作用机制是嵌入 DNA分子、 破坏核 酸结构从而抑制生物大分子的合成。 阿霉素主要用于治疗急性白血病、恶 性淋巴瘤、 肝癌、 肺癌、 胃癌、 食道癌、 宫颈癌、 膀胱癌、 睾丸癌、 曱状 腺癌、 软组织肿瘤、 骨肉瘤和神经母细胞癌等。 目前， 阿霉素最主要的临 床给药方式是静脉滴注，然而静脉滴注后药物会迅速分布全身，从而缺少 对肿瘤组织的靶向， 能达到肿瘤部位并发挥疗效的阿霉素比例很低，致使 其生物利用度不高， 效率低下。 因此， 临床需频繁给药， 从而对身体的正 常组织和器官造成较大的毒副作用。  Doxorubicin, also known as hydroxydaunorubicin, hydroxyd-dentamicin, doxorubicin or ADR, is a broad-spectrum anti-tumor drug of anthracycline antibiotics, a cell cycle non-specific drug with the strongest effect on S phase. It also has effects on ^1, G1 and G2 phases. Its mechanism of action is to embed DNA molecules and destroy nucleic acid structures to inhibit the synthesis of biological macromolecules. Doxorubicin is mainly used for the treatment of acute leukemia, malignant lymphoma, liver cancer, lung cancer, gastric cancer, esophageal cancer, cervical cancer, bladder cancer, testicular cancer, verrucous adenocarcinoma, soft tissue tumor, osteosarcoma and neuroblastoma. At present, the main clinical mode of administration of doxorubicin is intravenous infusion. However, after intravenous infusion, the drug is rapidly distributed throughout the body, thus lacking the target for tumor tissue, and the ratio of doxorubicin that can reach the tumor site and exert its therapeutic effect is very high. Low, resulting in low bioavailability and inefficiency. Therefore, clinical administration requires frequent administration, thereby causing a large toxic side effect on normal tissues and organs of the body.
为解决阿霉素在临床上存在的问题，阿霉素新剂型的开发一直是研究 者研究的热点。其中纳米尺度的药物传输系统的研究发展较为迅速， 包括 脂质体， 聚合物纳米颗粒， 胶束等。 其中， 由两亲性嵌段共聚物或接枝共 聚物制备而成的胶束成为药物载体的研究重点。胶束包载药物的方式主要 分为两种， 一种是物理包载， 一种是化学键合。 其中， 化学键合由于药物 包载方式更稳定、药物译放过程更持续以及药物译放时间更长久而获得了 更广泛的研究。  In order to solve the clinical problems of doxorubicin, the development of new doxorubicin dosage forms has been a hot spot for researchers. Among them, nanoscale drug delivery systems have been developed rapidly, including liposomes, polymer nanoparticles, and micelles. Among them, micelles prepared from amphiphilic block copolymers or graft copolymers have become the focus of research on drug carriers. There are two main ways in which micelles are loaded with drugs, one is physical encapsulation and the other is chemical bonding. Among them, chemical bonding has been more widely studied due to the more stable drug entrapment, the more sustained drug release process, and longer drug release times.
公开号为 CN101234204 A的中国专利公开了一种高分子阿霉素键合 药， 在聚乙二醇在溶剂和催化剂的作用下进行脂肪族环酯的开环聚合反 应， 得到聚乙二醇-聚乳酸嵌段共聚物； 再将该嵌段共聚物的端羟基转化 为端羧基，在缩合剂的作用下与阿霉素进行酰胺化反应，得到高分子阿霉 素键合药； 公开号为 CN101234205 A的中国专利公开了一种具有靶向功 能的高分子阿霉素键合药 , 由两种聚乙二醇-聚乳酸嵌段共聚物混合组装 而成， 第一种聚乙二醇 -聚乳酸嵌段共聚物的聚乳酸链端连接有阿霉素， 第二种聚乙二醇 -聚乳酸嵌段共聚物的聚乙二醇链端连接有乳糖， 阿霉素 受聚乙二醇和聚乳酸的双重保护， 具有緩释功能； 乳糖具有靶向功能， 能 够实现阿霉素的靶向输送。但是，上述两种阿霉素键合药均存在连结的化 学键过于稳定、载药量低、 阿霉素的译放缺乏智能性等问题。 Biomaterials ( Vol. 31 , pl360-1371 , 2010 )公开了一种在聚酰胺 -胺树枝状高分子的 表面氨基上键合聚乙二醇单曱醚和顺 -3-羧基戊烯二酸酐修饰的羧基化阿 霉素， 从而得到的高分子阿霉素键合药， 该键合药在酸性条件下， 在肿瘤 组织和细胞内能够快速译放，从而实现药物译放的智能性，但是该阿霉素 键合药所使用的载体材料为聚酰胺 -胺树枝状高分子， 其制备过程繁瑣、 生物相容性差， 不利于阿霉素键合药的实际应用。 The Chinese Patent Publication No. CN101234204 A discloses a polymer doxorubicin-bonding compound, which performs a ring-opening polymerization reaction of an aliphatic cyclic ester under the action of a solvent of a polyethylene glycol and a catalyst to obtain a polyethylene glycol- Polylactic acid block copolymer; the terminal hydroxyl group of the block copolymer is converted into a terminal carboxyl group, and amidation reaction with doxorubicin under the action of a condensing agent to obtain a polymer The Chinese Patent Publication No. CN101234205 A discloses a polymer-type doxorubicin-bonded drug having a targeting function, which is assembled by mixing two kinds of polyethylene glycol-polylactic acid block copolymers. The polylactic acid chain block copolymer of the first type of polyethylene glycol-polylactic acid block copolymer is linked with doxorubicin, and the polyethylene glycol chain end of the second polyethylene glycol-polylactic acid block copolymer is linked with lactose. Doxorubicin is protected by both polyethylene glycol and polylactic acid, and has a sustained release function; lactose has a targeting function and can achieve targeted delivery of doxorubicin. However, both of the above doxorubicin-bonding drugs have problems in that the linked chemical bonds are too stable, the drug loading is low, and the detoxification of doxorubicin is lacking in intelligence. Biomaterials (Vol. 31, pl360-1371, 2010) discloses a carboxyl group modified with polyethylene glycol monoterpene ether and cis-3-carboxypentadienoic anhydride on the surface amino group of a polyamide-amine dendrimer. The doxorubicin, thereby obtaining a polymer doxorubicin bonding drug, which can be quickly translated and released in tumor tissues and cells under acidic conditions, thereby realizing the intelligence of drug release, but the mold The carrier material used for the drug bonding is a polyamide-amine dendrimer, which is cumbersome to prepare and has poor biocompatibility, which is not conducive to the practical application of the doxorubicin bond.
发明内容 Summary of the invention
有鉴于此，本发明要解决的技术问题在于提供阿霉素键合药及其制备 方法， 本发明提供的阿霉素键合药制备过程简单、生物相容性较好、 可以 实现在肿瘤部位的聚集， 且具有 pH值响应性及主动靶向功能。  In view of the above, the technical problem to be solved by the present invention is to provide an adriamycin bonding drug and a preparation method thereof, and the doxorubicin bonding drug provided by the invention has simple preparation process, good biocompatibility, and can be realized in a tumor site. Aggregation with pH responsiveness and active targeting.
本发明提供了阿霉素键合药， 包括具有式（I )结构的第一重复单元、 式（II )结构的第二重复单元、 式（III ) 结构的第三重复单元和式（IV ) 结构的第四重复单元：  The present invention provides an adriamycin-bonding drug comprising a first repeating unit having the structure of the formula (I), a second repeating unit of the formula (II), a third repeating unit of the formula (III), and the formula (IV) The fourth repeating unit of the structure:
式（I ); Formula (I);
Figure imgf000004_0001
Figure imgf000004_0001
Figure imgf000005_0001
（IV);
Figure imgf000005_0001
(IV);
R选自式（1 ) 〜（4)结构中的一种: R is selected from one of the structures of the formula (1) to (4):
Figure imgf000005_0002
Figure imgf000005_0002
Lactonibic acid为乳糖酸， 其具有式 (5) 结构:
Figure imgf000006_0001
Lactonibic acid is lactobionic acid, which has the structure of formula (5):
Figure imgf000006_0001
(5)；  (5);
FA为叶酸， 其具有式（6)结构:  FA is folic acid, which has the structure of formula (6):
Figure imgf000006_0002
Figure imgf000006_0002
式（I)中， 所述第一重复单元的摩尔数 h、 第二重复单元的摩尔数 第三重复单元的摩尔数^ 第四重复单元的摩尔数 k与第一重复单元、 第 二重复单元、第三重复单元和第四重复单元的总摩尔数的比例满足以下条 件： 0.02≤h/(h+i+j+k)≤0.3; 0<i/(h+i+j+k)<0.3; 0≤j/(h+i+j+k)≤0.3。 In the formula (I), the number of moles of the first repeating unit, the number of moles of the second repeating unit, the number of moles of the third repeating unit, the number of moles of the fourth repeating unit, and the first repeating unit and the second repeating unit The ratio of the total number of moles of the third repeating unit and the fourth repeating unit satisfies the following condition: 0.02 ≤ h / (h + i + j + k) ≤ 0.3; 0 < i / (h + i + j + k) < 0.3; 0 ≤ j / (h + i + j + k) ≤ 0.3.
优 选 的 ， 0.15≤h/(h+i+j+k)≤0.3 , 0.10<i/(h+i+j+k)<0.20, Preferably, 0.15 ≤ h / (h + i + j + k) ≤ 0.3, 0.10 < i / (h + i + j + k) < 0.20,
0.10<j7(h+i+j+k)<0.20o 0.10<j7(h+i+j+k)<0.20o
优选的， 所述阿霉素键合药的数均分子量为 6000〜100000。  Preferably, the doxorubicin bond has a number average molecular weight of 6000 to 100,000.
本发明还提供了阿霉素键合药的制备方法， 包括以下步骤：  The invention also provides a preparation method of an adriamycin bonding drug, comprising the following steps:
葡聚糖、 羧基化阿霉素衍生物、 具有式（V)结构的乳糖酸和具有式 (VI)结构的叶酸与 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐和 4-二 曱氨基吡啶在有机溶剂中反应，得到阿霉素键合药，所述羧基化阿霉素衍 生物具有式（VII-a)、 式 (VII-b)、 式 ( VII-c )或式 ( VII-d )结构：
Figure imgf000007_0001
Dextran, carboxylated doxorubicin derivative, lactobionic acid having the structure of formula (V) and folic acid having the structure of formula (VI) and 1-(3-diaminoaminopropyl)-3-ethylcarbodiam The amine 'hydrochloride salt and 4-diguanidinoaminopyridine are reacted in an organic solvent to obtain a doxorubicin bond having the formula (VII-a), the formula (VII-b), Structure of formula (VII-c) or formula (VII-d):
Figure imgf000007_0001
优选的， 所述羧基化阿霉素衍生物、 乳糖酸、叶酸与葡聚糖主链上葡 萄糖重复单元的摩尔比为 （2〜30 ) ： ( 0〜30 )： ( 0〜30 )： 100。 Preferably, the molar ratio of the carboxylated doxorubicin derivative, lactobionic acid, folic acid to the glucose repeating unit on the dextran backbone is (2~30): (0~30): (0~30): 100 .
优选的， 所述羧基化阿霉素衍生物、 乳糖酸、叶酸与葡聚糖主链上葡 萄糖重复单元的摩尔比为 （8〜25 ) ： ( 5〜25 )： ( 5〜25 )： 100。  Preferably, the molar ratio of the carboxylated doxorubicin derivative, lactobionic acid, folic acid to the glucose repeating unit on the dextran backbone is (8~25): (5~25): (5~25): 100 .
优选的， 所述葡聚糖的分子量为 6000〜100000。  Preferably, the dextran has a molecular weight of 6000 to 100,000.
优选的， 所述有机溶剂为二曱基亚砜。  Preferably, the organic solvent is dimethyl sulfoxide.
优选的， 所述羧基化阿霉素衍生物按照以下方法制备：  Preferably, the carboxylated doxorubicin derivative is prepared according to the following method:
阿霉素 .盐酸盐、 酸酐和三乙胺在有机溶剂中反应， 得到羧基化的阿 霉素衍生物； 所述酸酐选自 1,2-二羧基环己烯酸酐、 丁二酸酐、 2,2,3,3- 四曱基丁二酸酐和顺 -3-羧基戊烯二酸酐中的一种。  Doxorubicin hydrochloride, anhydride and triethylamine are reacted in an organic solvent to obtain a carboxylated doxorubicin derivative; the anhydride is selected from the group consisting of 1,2-dicarboxycyclohexene anhydride, succinic anhydride, 2 One of 2,3,3-tetradecylsuccinic anhydride and cis-3-carboxypentenedioic anhydride.
优选的， 所述阿霉素 '盐酸盐、 酸酐和三乙胺的摩尔比为 1: ( 0.5〜2 )： ( 0.5-2 )。  Preferably, the molar ratio of the doxorubicin hydrochloride, anhydride and triethylamine is 1: (0.5~2): (0.5-2).
与现有技术相比，本发明以葡聚糖、具有式（II-a )、式（II-b )、式（II-c ) 或式（Π-d )结构的羧基化阿霉素衍生物、 乳糖酸和叶酸为原料， 使其发 生缩合反应得到阿霉素键合药。 其中， 乳糖酸和叶酸具有主动靶向功能， 本发明提供的阿霉素键合药可通过增强的渗透和滞留效应及主动靶向功 能实现药物在肿瘤部位的聚集。 同时，本发明提供的高分子阿霉素键合药 具有 pH响应性，通过酰胺键键合在高分子药物上的阿霉素在肿瘤组织或 肿瘤细胞内的 pH值条件下可以快速译放， 从而增强药物效果。 此外， 本 发明以葡聚糖为原料， 不仅具有良好的生物相容性， 而且简单易购， 有利 于阿霉素键合药的深入利用。 附图说明  Compared with the prior art, the present invention is derived from glucan, carboxylated doxorubicin having the structure of formula (II-a), formula (II-b), formula (II-c) or formula (Π-d). The substance, lactobionic acid and folic acid are used as raw materials to cause a condensation reaction to obtain an adriamycin bonding drug. Among them, lactobionic acid and folic acid have active targeting functions, and the doxorubicin bonding agent provided by the invention can achieve drug accumulation at the tumor site through enhanced penetration and retention effects and active targeting function. Meanwhile, the polymer doxorubicin-bonded drug provided by the invention has pH responsiveness, and the doxorubicin bonded to the polymer drug by the amide bond can be quickly translated under the pH condition of the tumor tissue or the tumor cell. Thereby enhancing the effect of the drug. Further, the present invention uses glucan as a raw material, and has not only good biocompatibility, but also simple and easy to purchase, which is advantageous for the in-depth utilization of doxorubicin-based drugs. DRAWINGS
图 1 为本发明实施例 1制备的羧基化阿霉素衍生物的核磁共振氢谱 图；  1 is a nuclear magnetic resonance spectrum of a carboxylated doxorubicin derivative prepared in Example 1 of the present invention;
图 2为本发明实施例 1制备的羧基化阿霉素衍生物的红外分析谱图； 图 3为本发明实施例 5制备的羧基化阿霉素衍生物的核磁共振氢谱 图。 具体实施方式 本发明提供了阿霉素键合药， 包括具有式（I)结构的第一重复单元、 式（II)结构的第二重复单元、 式（III) 结构的第三重复单元和式（IV) 结构的第四重复单元： 2 is an infrared analysis spectrum of a carboxylated doxorubicin derivative prepared in Example 1 of the present invention; and FIG. 3 is a nuclear magnetic resonance spectrum of a carboxylated doxorubicin derivative prepared in Example 5 of the present invention. detailed description The present invention provides an adriamycin-bonding drug comprising a first repeating unit having the structure of the formula (I), a second repeating unit of the formula (II), a third repeating unit of the formula (III), and the formula (IV) The fourth repeating unit of the structure:
Figure imgf000009_0001
Figure imgf000009_0001
R选自式（1)〜（4)结构中的一种:  R is selected from one of the structures of the formulae (1) to (4):
0 OH 0 0 OH 0
Figure imgf000009_0002
Figure imgf000010_0001
Figure imgf000009_0002
Figure imgf000010_0001
Lactonibicacid为乳糖酸， 具有式 (5)结构:  Lactonibicacid is lactobionic acid with the structure of formula (5):
Figure imgf000010_0002
Figure imgf000010_0002
(5);  (5);
FA为叶酸， 具有式（6)结构:  FA is folic acid and has the structure of formula (6):
H2N
Figure imgf000010_0003
(6); H 2 N
Figure imgf000010_0003
(6);
式（I)中， 所述第一重复单元的摩尔数 h、 第二重复单元的摩尔数 第三重复单元的摩尔数^ 第四重复单元的摩尔数 k和第一重复单元、 第 二重复单元、第三重复单元和第四重复单元的总摩尔数的比例满足以下条 件： 0.02<h/(h+i+j+k)<0.3; 0<i/(h+i+j+k)<0.3; 0<j/(h+i+j+k)<0.3; 优选的 满 足 以 下 条 件 ： 0.15<h/(h+i+j+k)<0.3 , 0.10<i/(h+i+j+k)<0.20, 0.10<j/(h+i+j+k)<0.20; 更优选的， 满足以下条件： 0.18<h/(h+i+j+k)<0.28, 0.12<i/(h+i+j+k)<0.18, 0.12<j/(h+i+j+k)<0.18。 In the formula (I), the number of moles of the first repeating unit, the number of moles of the second repeating unit, the number of moles of the third repeating unit, the number of moles of the fourth repeating unit, and the first repeating unit and the second repeating unit The ratio of the total number of moles of the third repeating unit and the fourth repeating unit satisfies the following condition: 0.02 < h / (h + i + j + k) <0.3; 0 < i / (h + i + j + k) <0.3;0<j/(h+i+j+k)<0.3; Preferably, the following conditions are satisfied: 0.15<h/(h+i+j+k)<0.3, 0.10<i/(h+i+j +k)<0.20, 0.10<j/(h+i+j+k)<0.20; More preferably, the following conditions are satisfied: 0.18<h/(h+i+j+k)<0.28, 0.12<i/(h+i+j +k) < 0.18, 0.12 < j / (h + i + j + k) < 0.18.
在本发明中， 在所述阿霉素键合药中， 其主链为葡聚糖， 葡萄糖重复 单元与阿霉素衍生物发生缩合， 得到具有式（I)结构的重复单元； 葡萄 糖重复单元与乳糖酸发生缩合， 得到具有式（II)结构的重复单元； 葡萄 糖重复单元与叶酸发生缩合， 得到具有式（III)结构的第三重复单元。 所 述阿霉素键合药的数均分子量为优选为 6000〜100000, 更优选为 7000-90000, 最优选为 8000〜80000。  In the present invention, in the doxorubicin-bonding drug, the main chain is dextran, and the glucose repeating unit is condensed with the doxorubicin derivative to obtain a repeating unit having the structure of the formula (I); the glucose repeating unit Condensation with lactobionic acid gives a repeating unit having the structure of formula (II); the glucose repeating unit is condensed with folic acid to obtain a third repeating unit having the structure of formula (III). The number average molecular weight of the doxorubicin bond is preferably from 6000 to 100,000, more preferably from 7,000 to 90,000, and most preferably from 8,000 to 80,000.
本发明还提供了阿霉素键合药的制备方法， 包括以下步骤： 葡聚糖、 羧基化阿霉素衍生物、 具有式（V)结构的乳糖酸和具有式 (VI)结构的叶酸与 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐和 4-二 曱氨基吡啶在有机溶剂中反应，得到阿霉素键合药，所述羧基化阿霉素衍 生物具有式（VII-a)、 式 (VII-b)、 式 ( VII-c )或式 ( VII-d )结构：  The invention also provides a preparation method of a doxorubicin bond, comprising the following steps: glucan, carboxylated doxorubicin derivative, lactobionic acid having the structure of formula (V) and folic acid having the structure of formula (VI) 1-(3-Diaminoaminopropyl)-3-ethylcarbodiimide' hydrochloride and 4-diguanidinoaminopyridine are reacted in an organic solvent to obtain an adriamycin-binding drug, the carboxylated A The derivative of the formula has the structure of the formula (VII-a), the formula (VII-b), the formula (VII-c) or the formula (VII-d):
Figure imgf000011_0001
Figure imgf000011_0001
(V); (V);
Figure imgf000011_0002
Figure imgf000011_0002
(VI)；
Figure imgf000012_0001
(VI);
Figure imgf000012_0001
本发明首先将葡聚糖、 羧基化阿霉素衍生物、 具有式（V)结构的乳 糖酸和具有式（VI) 结构的叶酸加入到有机溶剂中， 再加入 1-(3-二曱氨 基丙基 )-3-乙基碳二亚胺.盐酸盐和 4-二曱氨基吡啶进行反应，得到阿霉素 键合药。 在本发明中， 所述羧基化阿霉素衍生物、 乳糖酸、 叶酸与葡聚糖 主链上葡萄糖重复单元的摩尔比优选为 （2〜30)： (0〜30)： (0〜30)： 100; 更优选为 （8〜25) ： (5〜25)： (5〜25)： 100; 所述 1-(3-二曱氨基丙基 )-3- 乙基碳二亚胺.盐酸盐的摩尔数与所述羧基化阿霉素衍生物、 乳糖酸和叶 酸的摩尔数的总和的比例优选为 1: (4〜6)； 更优选为 1: (4.5〜5.5)； 所 述 4-二曱氨基吡啶的摩尔数与所述羧基化阿霉素衍生物、 乳糖酸和叶酸 的摩尔数的总和的比例优选为 1: ( 1〜3), 更优选为 1: ( 1.5〜2.5)； 所述 葡聚糖的分子量优选为 6000〜100000, 更优选为 7000〜90000, 最优选为 8000-80000; 所述有机溶剂优选为二曱基亚砜； 所述反应优选在搅拌的 条件下进行。 所述反应温度优选为室温； 所述反应时间优选为 3〜5 天， 更优选为 3.5〜4天。  The invention firstly adds dextran, carboxylated doxorubicin derivative, lactobionic acid having the structure of formula (V) and folic acid having the structure of formula (VI) to an organic solvent, and then adding 1-(3-diaminoamino group). The propyl)-3-ethylcarbodiimide hydrochloride is reacted with 4-diaminopyridine to give an adriamycin bond. In the present invention, the molar ratio of the carboxylated doxorubicin derivative, lactobionic acid, folic acid to the glucose repeating unit on the dextran main chain is preferably (2 to 30): (0 to 30): (0 to 30) 100: More preferably (8~25): (5~25): (5~25): 100; the 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide. The ratio of the number of moles of the hydrochloride to the sum of the moles of the carboxylated doxorubicin derivative, lactobionic acid and folic acid is preferably 1: (4 to 6); more preferably 1: (4.5 to 5.5); The ratio of the number of moles of 4-diaminopyridine to the sum of the moles of the carboxylated doxorubicin derivative, lactobionic acid and folic acid is preferably 1: (1 to 3), more preferably 1: (1.5~ 2.5); the molecular weight of the glucan is preferably from 6000 to 100000, more preferably from 7000 to 90,000, most preferably from 8,000 to 80,000; the organic solvent is preferably dimercaptosulfoxide; the reaction is preferably under stirring conditions Go on. The reaction temperature is preferably room temperature; the reaction time is preferably from 3 to 5 days, more preferably from 3.5 to 4 days.
反应完毕后，将反应液装入透析袋中透析 ,冻干后得到阿霉素键合药。 所述透析袋的截留分子量优选为 3500Dalton;所述透析的时间优选为 3〜5 天， 更优选为 3.5〜4天； 所述透析过程中优选每隔 3〜5小时换水一次， 更 优选为 3.5〜4.5小时换水一次； 本发明对于冷冻干燥并无限制， 本领域技 术人员熟知的冷冻干燥即可。 After the reaction is completed, the reaction solution is dialyzed into a dialysis bag, and lyophilized to obtain an adriamycin-binding drug. The molecular weight cut off of the dialysis bag is preferably 3,500 Dalton; the time of the dialysis is preferably 3 to 5 More preferably, it is 3.5 to 4 days; in the dialysis process, water is preferably changed every 3 to 5 hours, more preferably 3.5 to 4.5 hours; the present invention is not limited to freeze drying, and those skilled in the art It is well known to freeze and dry.
本发明对所述葡聚糖的来源没有限制， 市场上购买即可。  The present invention has no limitation on the source of the glucan, and it is commercially available.
本发明对所述乳糖酸和叶酸的来源没有限制， 市场上购买即可。  The present invention has no limitation on the source of the lactobionic acid and folic acid, and is commercially available.
所述羧基化阿霉素衍生物具有式（VII-a )、 式（VII-b )、 式（VII-c ) 或式（Vll-d )结构， 优选按照以下方法制备：  The carboxylated doxorubicin derivative has a structure of the formula (VII-a), the formula (VII-b), the formula (VII-c) or the formula (Vll-d), preferably prepared according to the following method:
阿霉素 .盐酸盐、 酸酐和三乙胺在有机溶剂中发生反应， 得到羧基化 的阿霉素衍生物， 所述酸酐为 1,2-二羧基环己烯酸酐、 丁二酸酐、 2,2,3,3- 四曱基丁二酸酐或顺 -3-羧基戊烯二酸酐。  The doxorubicin hydrochloride, the acid anhydride and the triethylamine are reacted in an organic solvent to obtain a carboxylated doxorubicin derivative, which is 1,2-dicarboxycyclohexene anhydride, succinic anhydride, 2 , 2,3,3-tetradecyl succinic anhydride or cis-3-carboxypentene dianhydride.
在无水条件下， 将阿霉素.盐酸盐、 酸酐和三乙胺溶解于有机溶剂中， 阿霉素 ·盐酸盐和酸酐发生反应， 得到羧基化的阿霉素衍生物。 所述有机 溶剂优选为 Ν,Ν-二曱基曱酰胺； 所述阿霉素 '盐酸盐、 酸酐和三乙胺的摩 尔比优选为 1: ( 0.5〜2 )： ( 0.5〜2 ), 更优选为 1: ( 1-1.5 )： ( 1〜1.5 )。 所述 阿霉素 ·盐酸盐和酸酐优选在搅拌的条件下发生反应， 所述反应的温度优 选为 10°C〜40°C , 更优选为 15°C〜35°C , 最优选为 20°C〜30°C , 时间优选 为 101！〜 50h, 更优选为 15h〜40h, 最优选为 20h〜30h。 反应结束后， 将得 到的反应混合物倒入 20倍体积的乙酸乙酯中稀译， 然后用饱和氯化钠水 溶液洗涤、 干燥、 过滤、 浓缩后得到羧基化阿霉素衍生物。  Under anhydrous conditions, doxorubicin hydrochloride, anhydride and triethylamine are dissolved in an organic solvent, and doxorubicin hydrochloride and an acid anhydride are reacted to obtain a carboxylated doxorubicin derivative. The organic solvent is preferably ruthenium, osmium-dimercaptoamide; the molar ratio of the doxorubicin hydrochloride, anhydride and triethylamine is preferably 1: (0.5~2): (0.5~2), More preferably, it is 1: (1-1.5): (1 to 1.5). The doxorubicin hydrochloride and the acid anhydride are preferably reacted under stirring, and the temperature of the reaction is preferably from 10 ° C to 40 ° C, more preferably from 15 ° C to 35 ° C, and most preferably 20 °C~30°C, the time is preferably 101! 〜50h, more preferably 15h~40h, most preferably 20h~30h. After completion of the reaction, the obtained reaction mixture was poured into 20 volumes of ethyl acetate, and then washed with a saturated aqueous solution of sodium chloride, dried, filtered, and concentrated to give a carboxylated doxorubicin derivative.
在本发明中， 所述酸酐为 1,2-二羧基环己烯酸酐、 丁二酸酐、 2,2,3,3- 四曱基丁二酸酐或顺 -3-羧基戊烯二酸酐，当所述酸酐为 1,2-二羧基环己烯 酸酐时， 得到的羧基化阿霉素衍生物具有式（VII -a ) 结构； 当所述酸酐 为丁二酸酐时， 得到的羧基化阿霉素衍生物具有式（VII -b ) 结构； 当所 述酸酐为 2,2,3,3-四曱基丁二酸酐时， 得到的羧基化阿霉素衍生物具有式 ( VII -c )结构； 当所述酸酐为顺 -3-羧基戊烯二酸酐时， 得到的羧基化阿 霉素衍生物具有式（Vll -d ) 结构。  In the present invention, the acid anhydride is 1,2-dicarboxycyclohexene anhydride, succinic anhydride, 2,2,3,3-tetradecyl succinic anhydride or cis-3-carboxypentene dianhydride. When the acid anhydride is 1,2-dicarboxycyclohexene anhydride, the obtained carboxylated doxorubicin derivative has a structure of the formula (VII-a); when the acid anhydride is succinic anhydride, the obtained carbamate The derivative of the formula has the structure of the formula (VII-b); when the anhydride is 2,2,3,3-tetradecylsuccinic anhydride, the obtained carboxylated doxorubicin derivative has the structure of the formula (VII-c) When the acid anhydride is cis-3-carboxypentadienoic anhydride, the obtained carboxylated doxorubicin derivative has a structure of the formula (V11-d).
本发明以葡聚糖、 具有式（II-a )、 式（II-b )、 式 ( II-c )或式 ( ΙΙ-d ) 结构的羧基化阿霉素衍生物、乳糖酸和叶酸为原料，使其发生缩合反应得 到阿霉素键合药。 其中， 乳糖酸和叶酸具有主动靶向功能， 本发明提供的 阿霉素键合药可通过增强的渗透和滞留效应及主动靶向功能实现药物在 肿瘤部位的聚集。 进一步的， 本发明提供的高分子阿霉素键合药具有 pH 响应性，通过酰胺键键合在高分子药物上的阿霉素在肿瘤组织或肿瘤细胞 内的 pH值条件下可以快速译放， 从而增强药物效果。 此外， 本发明以葡 聚糖为原料， 不仅具有良好的生物相容性， 而且简单易购， 有利于阿霉素 键合药的深入利用。 The present invention uses dextran, a carboxylated doxorubicin derivative having the structure of formula (II-a), formula (II-b), formula (II-c ) or formula (II-d), lactobionic acid and folic acid. The raw material is subjected to a condensation reaction to obtain an adriamycin-bonding drug. Among them, lactobionic acid and folic acid have active targeting functions, and the present invention provides Adriamycin-based drugs can achieve drug accumulation at the tumor site through enhanced penetration and retention effects and active targeting. Further, the polymer doxorubicin-bonded drug provided by the invention has pH responsiveness, and the doxorubicin bonded to the polymer drug by the amide bond can be quickly released under the pH condition of the tumor tissue or the tumor cell. , thereby enhancing the effect of the drug. In addition, the invention uses dextran as a raw material, has not only good biocompatibility, but also is easy to purchase, which is beneficial to the in-depth utilization of the doxorubicin bond.
为了进一步说明本发明，以下结合实施例对本发明提供的阿霉素键合 药及其制备方法进行详细描述。  In order to further illustrate the present invention, the doxorubicin-bonded drug provided by the present invention and a method for producing the same are described in detail below with reference to examples.
实施例 1  Example 1
将 580.0mg ( O.OOlmol ) 阿霉素 '盐酸盐、 156.1mg ( O.OOlmol )顺 -3- 氣基戊烯二酸酐和 101.2mg ( O.OOlmol ) 三乙胺置于干燥的反应瓶中， 加 入 5mL无水 Ν,Ν-二曱基曱酰胺溶解，在 25°C、搅拌子搅拌的条件下反应 24h , 反应结束后 , 将得到的反应混合液倒入 1 OOmL的乙酸乙酯中稀释 , 用饱和氯化钠水溶液洗涤、干燥、过滤、浓缩得到羧基化的阿霉素衍生物。  580.0 mg (0.0 billion mol) of doxorubicin 'hydrochloride, 156.1 mg (0.0 billion mol) of cis-3-pentylidene dianhydride and 101.2 mg (0.0 billion mol) of triethylamine were placed in a dry reaction flask Add 5 mL of anhydrous hydrazine, dissolve in hydrazine-dihydrazinamide, and react at 25 ° C with stirring under stirring for 24 h. After the reaction is over, pour the obtained reaction mixture into 100 mL of ethyl acetate. Dilute, wash with saturated aqueous sodium chloride solution, dry, filter and concentrate to give a carboxylated doxorubicin derivative.
对所述羧基化的阿霉素衍生物进行核磁共振分析， 结果参见图 1 , 图 Nuclear magnetic resonance analysis of the carboxylated doxorubicin derivative, the results are shown in Figure 1, Figure
1为本发明实施例 1制备的羧基化阿霉素衍生物的核磁共振氢谱图；对所 述羧基化的阿霉素衍生物进行红外分析， 结果参见图 2, 图 2为本发明实 施例 1制备的羧基化阿霉素衍生物的红外分析谱图， 由图 1和图 2可知， 本发明实施例制备的羧基化阿霉素衍生物具有式（Π-d )结构。 1 is a nuclear magnetic resonance spectrum of the carboxylated doxorubicin derivative prepared in Example 1 of the present invention; infrared analysis of the carboxylated doxorubicin derivative, the results are shown in FIG. 2, and FIG. 2 is an embodiment of the present invention. An infrared analysis spectrum of the prepared carboxylated doxorubicin derivative, as shown in Fig. 1 and Fig. 2, the carboxylated doxorubicin derivative prepared in the examples of the present invention has a structure of the formula (Π-d).
实施例 2  Example 2
将 580.0mg ( O.OOlmol ) 阿霉素 '盐酸盐、 152.2mg ( O.OOlmol ) 1,2- 二羧基环己烯酸酐和 101.2mg ( O.OOlmol ) 三乙胺置于干燥反应瓶中， 加 入 5mL无水 Ν,Ν-二曱基曱酰胺溶解，在 25°C、搅拌子搅拌的条件下反应 24h , 反应结束后 , 将得到的反应混合液倒入 1 OOmL的乙酸乙酯中稀释 , 用饱和氯化钠水溶液洗涤、 干燥、 过滤、 浓缩后得到羧基化的阿霉素衍生 物。  580.0 mg (0.0 billion mol) of doxorubicin hydrochloride, 152.2 mg (0.0 billion mol) of 1,2-dicarboxycyclohexene anhydride and 101.2 mg (0.0 billion mol) of triethylamine were placed in a dry reaction flask. Add 5 mL of anhydrous hydrazine, dissolve in hydrazine-dihydrazinamide, and react at 25 ° C with stirring under stirring for 24 h. After the reaction is over, pour the obtained reaction mixture into 100 mL of ethyl acetate to dilute. It is washed with a saturated aqueous solution of sodium chloride, dried, filtered, and concentrated to give a carboxylated doxorubicin derivative.
对所述羧基化的阿霉素衍生物进行核磁共振分析和红外分析，结果表 明， 本发明实施例 2制备的羧基化阿霉素衍生物具有式（Π-a )结构。  The nuclear magnetic resonance analysis and infrared analysis of the carboxylated doxorubicin derivative showed that the carboxylated doxorubicin derivative prepared in Example 2 of the present invention had a structure of the formula (Π-a).
实施例 3 将 580.0mg ( O.OOlmol ) 阿霉素 '盐酸盐、 126.1mg ( O.OOlmol ) 丁二 酸酐和 101.2mg ( O.OOlmol )三乙胺置于干燥的反应瓶中， 加入 5mL无水 Ν,Ν-二曱基曱酰胺溶解， 在 25°C、 搅拌子搅拌的条件下反应 24h, 反应 结束后， 将得到的反应混合液倒入 lOOmL的乙酸乙酯中稀释， 用饱和氯 化钠水溶液洗涤、 干燥、 过滤、 浓缩得到羧基化的阿霉素衍生物。 Example 3 580.0 mg (0.0 billion mol) of doxorubicin hydrochloride, 126.1 mg (0.0 billion mol) of succinic anhydride and 101.2 mg (0.0 billion mol) of triethylamine were placed in a dry reaction flask, and 5 mL of anhydrous hydrazine was added. , Ν-dimercaptoamide is dissolved, and reacted at 25 ° C with stirring under stirring for 24 h. After the reaction is completed, the obtained reaction mixture is poured into 100 mL of ethyl acetate and diluted with a saturated aqueous solution of sodium chloride. Washing, drying, filtration, and concentration give a carboxylated doxorubicin derivative.
对所述羧基化的阿霉素衍生物进行核磁共振分析和红外分析，结果表 明， 本发明实施例 3制备的羧基化阿霉素衍生物具有式（Π-b )结构。  The nuclear magnetic resonance analysis and infrared analysis of the carboxylated doxorubicin derivative showed that the carboxylated doxorubicin derivative prepared in Example 3 of the present invention had a structure of the formula (Π-b).
实施例 4  Example 4
将 580.0mg ( O.OOlmol )阿霉素 '盐酸盐， 156.2mg ( O.OOlmol ) 2,2,3,3- 四曱基丁二酸酐和 101.2mg ( O.OOlmol )三乙胺置于干燥的反应瓶中， 加 入 5mL无水 Ν,Ν-二曱基曱酰胺溶解，在 25°C ,搅拌子搅拌的条件下反应 24h , 反应结束后 , 把得到的反应混合液倒入 1 OOmL的乙酸乙酯中稀释 , 用饱和氯化钠水溶液洗涤、干燥、过滤、浓缩得到羧基化的阿霉素衍生物。  580.0 mg (0.0 billion mol) of doxorubicin hydrochloride, 156.2 mg (0.0 billion mol) of 2,2,3,3-tetradecylsuccinic anhydride and 101.2 mg (0.0 billion mol) of triethylamine were placed In a dry reaction flask, 5 mL of anhydrous hydrazine and hydrazine-dihydrazinamide were added and dissolved at 25 ° C with stirring under stirring for 24 h. After the reaction was completed, the obtained reaction mixture was poured into 100 mL. It was diluted with ethyl acetate, washed with a saturated aqueous solution of sodium chloride, dried, filtered, and concentrated to give a crystallized doxorubicin derivative.
对所述羧基化的阿霉素衍生物进行核磁共振分析和红外分析，结果表 明， 本发明实施例 4制备的羧基化阿霉素衍生物具有式（ II-c )结构。  The nuclear magnetic resonance analysis and infrared analysis of the carboxylated doxorubicin derivative showed that the carboxylated doxorubicin derivative prepared in Example 4 of the present invention had the structure of the formula (II-c).
实施例 5〜7  Example 5~7
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )，放入 3个反应瓶中， 分别加入 0.01447g ( 0.0000207mol )实施例 4制备的羧基化的阿霉素衍生 物， 0.02284g、 0.06853和 0.1371g的叶酸， 0.13890g、 0.33733g和 0.6350g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.00885g、 0.02150g 和 0.04046g的 4-二曱氨基吡啶， 混合均匀后， 再分别加入 25mL二曱基亚 砜溶解反应体系， 室温避光搅拌条件下反应 3天，反应结束后， 将得到的 反应混合液用截留分子量为 3500 Dalton的透析袋透析 3天， 每隔 5 h换 水一次， 将反应液冻干， 分别得到阿霉素键合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) were placed in three reaction flasks, and 0.01447 g (0.0000 207 mol) of the carboxylated doxorubicin derivative prepared in Example 4, 0.02284 g, 0.06853 and 0.1371 g of folic acid, 0.13890 g, 0.33733 g and 0.6350 g of 1-(3-diamidinopropyl)-3-ethylcarbodiimide' hydrochloride, 0.00885 g, 0.02150 g and 0.04046 g After the mixture was uniformly mixed, 25 mL of dimethylsulfoxide was added to dissolve the reaction system, and the reaction was carried out for 3 days at room temperature under light agitation. After the reaction was completed, the obtained reaction mixture was subjected to a molecular weight cutoff of 3,500 Dalton. The dialysis bag was dialyzed for 3 days, the water was changed every 5 hours, and the reaction solution was lyophilized to obtain an adriamycin-binding drug.
对实施例 5制备的阿霉素键合药进行核磁共振分析， 结果见图 3 , 图 The nuclear magnetic resonance analysis of the doxorubicin-bonded drug prepared in Example 5 was carried out, and the results are shown in Fig. 3.
3为本发明实施例 5制备的阿霉素键合药的核磁共振氢谱图。 结果如下： 8.57ppm、 7,95ppm、 7.61ppm 为 -FA (叶酸） 的特征峰； 4.0〜5.0ppm、 3.8〜3.0ppm为葡聚糖的特征峰； 3.3ppm为水的特征峰； 4.0ppm为^^化 阿霉素衍生物的特征峰； 2.5ppm为氘代 DMSO的特征峰。 结果表明， 本 发明实施例 5制备的阿霉素键合药具有式（I )结构， 其中， h、 j、 k取值 见表 1。 3 is a nuclear magnetic resonance spectrum of the doxorubicin-bonded drug prepared in Example 5 of the present invention. The results are as follows: 8.57 ppm, 7,95 ppm, 7.61 ppm are characteristic peaks of -FA (folic acid); 4.0 to 5.0 ppm, 3.8 to 3.0 ppm are characteristic peaks of dextran; 3.3 ppm is characteristic peak of water; 4.0 ppm is ^^ Characteristic peak of doxorubicin derivative; 2.5ppm is a characteristic peak of deuterated DMSO. The result shows that this The doxorubicin-bonded drug prepared in Inventive Example 5 has the structure of the formula (I), wherein the values of h, j, and k are shown in Table 1.
对实施例 5制备的阿霉素键合药进行红外分析，结果表明，本发明实 施例 5制备的阿霉素键合药具有式（I )结构。  Infrared analysis of the doxorubicin-bonded drug prepared in Example 5 revealed that the doxorubicin-bonded drug prepared in Example 5 of the present invention had the structure of the formula (I).
分别对实施例 6和实施例 7制备的阿霉素键合药进行核磁共振分析和 红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 j、 k取值见表 1。  The nuclear magnetic resonance analysis and infrared analysis of the doxorubicin-bonded drugs prepared in Example 6 and Example 7 respectively showed that they all have the structure of the formula (I), wherein the values of h, j, and k are shown in Table 1.
表 1本发明实施例 5〜7制备得到的高分子阿霉素键合药 h、 j、 k取值 实施例 Mn (g mol"1) h j k Table 1 Examples of the polymer doxorubicin bonding drugs h, j, k prepared in Examples 5 to 7 of the present invention. Example M n (g mol" 1 ) hjk
5 6000 0.02 0.05 0.93 5 6000 0.02 0.05 0.93
6 6000 0.02 0.15 0.836 6000 0.02 0.15 0.83
7 6000 0.02 0.30 0.68 实施例 8〜10 7 6000 0.02 0.30 0.68 Example 8~10
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )和 3份 0.00133g ( 0.0000207mol )实施例 2制备的羧基化的阿霉素衍生物，放入 3个反应 瓶中， 再分别加入 0.01854g、 0.05563g和 0.11125g的乳糖酸， 0.13890g、 0.33731g 和 0.63325g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.00885g、 0.02150g和 0.04046g的 4-二曱氨基吡啶， 混合均匀后, 再分 别加入 25mL二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3 天, 反应结束后， 将得到的反应混合液用截留分子量为 3500 Dalton的透析袋 透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键 合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) and 3 parts of 0.00133 g (0.0000207 mol) of the carboxylated doxorubicin derivative prepared in Example 2 were placed in 3 reaction bottles, and then separately added. 0.01854g, 0.05563g and 0.11125g of lactobionic acid, 0.13890g, 0.33731g and 0.63325g of 1-(3-diamidinopropyl)-3-ethylcarbodiimide' hydrochloride, 0.00885g, 0.02150 g and 0.04046g of 4-diaminopyridine, after mixing uniformly, add 25mL of dimercaptosulfoxide to dissolve the reaction system, react at room temperature for 3 days under light agitation, and after the reaction is finished, use the obtained reaction mixture. The dialysis bag with a molecular weight cut off of 3500 Dalton was dialyzed for 3 days, and the water was changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 8〜10制备的高分子阿霉素键合药进行核磁共振 分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取值 见表 2。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 8 to 10 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 2. .
表 2本发明实施例 8〜10制备 的高分子阿霉素键合药 h、 i、 k取
Figure imgf000016_0001
Table 2 The polymer doxorubicin bonding drugs h, i, k prepared in Examples 8 to 10 of the present invention were taken.
Figure imgf000016_0001
实施例 Mn (g mol"1) h i k Example M n (g mol" 1 ) hik
8 6000 0.02 0.05 0.93 8 6000 0.02 0.05 0.93
9 6000 0.02 0.15 0.83 10 6000 O02 030 0.68 实施例 11〜13 9 6000 0.02 0.15 0.83 10 6000 O02 030 0.68 Examples 11 to 13
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )和 3份 0.06667g ( 0.00010327mol ) 实施例 2制备的羧基化的阿霉素衍生物， 放入 3个反 应瓶中，再分别加入 0.01854g、 0.05563g和 0.11125g的乳糖酸， 0.29698g、 0.49497g 和 0.79197g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.01893g、 0.03154g和 0.05047g的 4-二曱氨基吡啶， 混合均匀后， 再分 别加入 25mL二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3 天, 反应结束后， 将得到的反应混合液用截留分子量为 3500 Dalton的透析袋 透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键 合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) and 3 parts of 0.06667 g (0.00010327 mol) of the carboxylated doxorubicin derivative prepared in Example 2 were placed in 3 reaction bottles and separately added. 0.01854g, 0.05563g and 0.11125g of lactobionic acid, 0.29698g, 0.49497g and 0.79197g of 1-(3-diamidinopropyl)-3-ethylcarbodiimide' hydrochloride, 0.01893g, 0.03154 g and 0.05047 g of 4-diaminopyridine, after mixing uniformly, add 25 mL of dimercaptosulfoxide to dissolve the reaction system, and react at room temperature for 3 days under light agitation. After the reaction is finished, the obtained reaction mixture is used. The dialysis bag with a molecular weight cut off of 3500 Dalton was dialyzed for 3 days, and the water was changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 11〜13 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取 值见表 3。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 11 to 13 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 3. .
表 3 本发明实施例 11〜13制备得到的高分子阿霉素键合药 h、 i、 k 取值  Table 3 The polymer doxorubicin bonding drugs prepared in Examples 11 to 13 of the present invention have values of h, i, and k
实施例 Mn (g ηιοΓ1) h i k Example M n (g ηιοΓ 1 ) hik
11 6000 0.10 0.05 0.85 11 6000 0.10 0.05 0.85
12 6000 0.10 0.15 0.7512 6000 0.10 0.15 0.75
13 6000 0.10 0.30 0.60 实施例 14〜16 13 6000 0.10 0.30 0.60 Example 14~16
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )和 3份 0.0133g ( 0.0002065mol )实施例 2制备的羧基化的阿霉素衍生物，放入 3个反应 瓶中， 再分别加入 0.01854g、 0.05563g和 0.11125g的乳糖酸， 0.49488g、 0.69283g 和 0.98975g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.03154g、 0.04415g和 0.06307g的 4-二曱氨基吡啶， 混合均匀后， 再分 别加入 25mL二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton的透 析袋透析 3天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉 素键合药。 3 parts of 0.16768 g of dextran (number average molecular weight of 6000) and 3 parts of 0.01133 g (0.0002065 mol) of the carboxylated doxorubicin derivative prepared in Example 2 were placed in 3 reaction bottles, and then separately added. 0.01854g, 0.05563g and 0.11125g of lactobionic acid, 0.49488g, 0.69283g and 0.98975g of 1-(3-diaminopropyl)-3-ethylcarbodiimide' hydrochloride, 0.03154g, 0.04415 g and 0.06307g of 4-diaminopyridine, after mixing uniformly, add 25mL of dimercaptosulfoxide to dissolve the reaction system, react at room temperature for 3 days under light agitation, and after the reaction is finished, separately obtain the reaction mixture. Dialysis was carried out for 3 days in a dialysis bag with a molecular weight cut off of 3,500 Dalton, and the water was changed every 5 hours, and the reaction solution was lyophilized to obtain a polymer mold. Prime bonding drug.
分别对本发明实施例 11〜13 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取 值见表 4。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 11 to 13 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 4. .
表 4 本发明实施例 14〜16制备得到的高分子阿霉素键合药 h、 i、 k 取值  Table 4 The polymer doxorubicin bonding drugs prepared in Examples 14 to 16 of the present invention have values of h, i, and k
实施例 Mn (g mol"1) h i k Example M n (g mol" 1 ) hik
14 6000 0.20 0.05 0.75 14 6000 0.20 0.05 0.75
15 6000 0.20 0.15 0.6515 6000 0.20 0.15 0.65
16 6000 0.20 0.30 0.50 实施例 17〜19 16 6000 0.20 0.30 0.50 Example 17~19
将 3 份 0.16768g 葡聚糖（数均分子量为 6000 )和 3 份 0.20001g (0.0003098mol)实施例 2制备的羧基化的阿霉素衍生物， 放入 3个反应瓶 中， 再分别加入 0.01826g、 0.05478g和 0.10956g的乳糖酸， 0.69294g、 0.89092g 和 1.18797g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.04416g、 0.05677g和 0.06308g的 4-二曱氨基吡啶， 混合均匀后， 再分 别加入 25mL二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3天, 反应结束后分别将得到的反应混合液用截留分子量为 3500 Dalton的透析 袋透析 3天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素 键合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) and 3 parts of 0.20001 g (0.0003098 mol) of the carboxylated doxorubicin derivatives prepared in Example 2 were placed in 3 reaction bottles, and then added to 0.01826 respectively. g, 0.05478 g and 0.10956 g of lactobionic acid, 0.69294 g, 0.89092 g and 1.18797 g of 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide' hydrochloride, 0.04416 g, 0.05677 g After mixing with 0.06308 g of 4-diaminopyridine, the reaction system was dissolved in 25 mL of dimercaptosulfoxide, and reacted at room temperature for 3 days under light agitation. After the reaction was completed, the obtained reaction mixture was separately retained. The dialysis bag having a molecular weight of 3,500 Dalton was dialyzed for 3 days, and the water was changed every 5 hours, and the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 17〜19制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取 值见表 5。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 17 to 19 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 5. .
表 5 本发明实施例 17〜19制备得到的高分子阿霉素键合药 h、 i、 k 取值  Table 5 The polymer doxorubicin bonding drugs prepared in Examples 17 to 19 of the present invention have values of h, i, and k
实施例 Mn (g mol"1) h i k Example M n (g mol" 1 ) hik
17 6000 0.30 0.05 0.65 17 6000 0.30 0.05 0.65
18 6000 0.30 0.15 0.5518 6000 0.30 0.15 0.55
19 6000 0.30 0.30 0.40 实施例 20〜22 19 6000 0.30 0.30 0.40 Example 20~22
将 3 份 0.16768g 葡聚糖（数均分子量为 6000 )和 3 份 0.00133g ( 0.0000207mol )实施例 2制备的羧基化的阿霉素衍生物，放入 3个反应 瓶中， 再分别加入 0.02284g、 0.06853g和 0.1371g 的叶酸， 0.13890g、 0.33733g 和 0.63498g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.00885g、 0.02150g和 0.04046g的 4-二曱氨基吡啶， 混合均匀后, 再分 别加入 25mL二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton的透 析袋透析 3天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉 素键合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) and 3 parts of 0.00133 g (0.0000207 mol) of the carboxylated doxorubicin derivative prepared in Example 2 were placed in 3 reaction bottles, and then added to 0.02284 g, 0.06853g and 0.1371g of folic acid, 0.13890g, 0.33733g and 0.63498g of 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide' hydrochloride, 0.00885g, 0.02150g and 0.04046g of 4-diaminopyridine, after mixing uniformly, add 25mL of dimercaptosulfoxide to dissolve the reaction system, react at room temperature for 3 days under light agitation, and after the reaction is finished, separate the obtained reaction mixture. The dialysis bag having a molecular weight of 3,500 Dalton was dialyzed for 3 days, and the water was changed every 5 hours, and the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 20〜22制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 j、 k取 值见表 6。  The polymer magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 20 to 22 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, j and k are shown in Table 6. .
表 6 本发明实施例 20〜22制备得到的高分子阿霉素键合药 h、 j、 k 取值  Table 6 In the present invention, the polymer doxorubicin bonding drugs prepared in the preparation of 20 to 22 have values of h, j, and k.
实施例 Mn (g ηιοΓ1) h j k Example M n (g ηιοΓ 1 ) hjk
20 6000 0.02 0.05 0.93 20 6000 0.02 0.05 0.93
21 6000 0.02 0.15 0.8321 6000 0.02 0.15 0.83
22 6000 0.02 0.30 0.68 实施例 23〜25 22 6000 0.02 0.30 0.68 Example 23~25
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )和 3份 0.06667g ( 0.00010327mol ) 实施例 2制备的羧基化的阿霉素衍生物， 放入 3个反 应瓶中， 再分别加入 0.02284g、 0.06853g和 0.1371g的叶酸， 0.29698g、 0.49497g 和 0.79197g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.01892g、 0.03154g和 0.05047g的 4-二曱氨基吡啶， 混合均匀后， 再分 别加入 25mL二曱基亚砜溶解反应体系，室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton的透 析袋透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉 素键合药。 分别对本发明实施例 23〜25 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 j、 k取 值见表 7。 3 parts of 0.16768 g of dextran (number average molecular weight of 6000) and 3 parts of 0.06667 g (0.00010327 mol) of the carboxylated doxorubicin derivative prepared in Example 2 were placed in 3 reaction bottles, and then separately added. 0.02284 g, 0.06853 g and 0.1371 g of folic acid, 0.29698 g, 0.49497 g and 0.79197 g of 1-(3-diamidinopropyl)-3-ethylcarbodiimide' hydrochloride, 0.01892 g, 0.03154 g After mixing with 0.05047 g of 4-diaminopyridine, the reaction system was dissolved in 25 mL of dimercaptosulfoxide, and reacted at room temperature for 3 days under light agitation. After the reaction was completed, the obtained reaction mixture was respectively used. The dialysis bag with a molecular weight cut off of 3500 Dalton was dialyzed for 3 days, and the water was changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond. The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 23 to 25 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, j and k are shown in Table 7. .
表 7本发明实施例 23〜25制备得到的高分子阿霉素键合药 h、 j、 k取 值  Table 7 Inventive Example 23 to 25 prepared polymer doxorubicin bonding drugs h, j, k take values
实施例 Mn (g ηιοΓ1) h j k Example M n (g ηιοΓ 1 ) hjk
23 6000 0.10 0.05 0.85 23 6000 0.10 0.05 0.85
24 6000 0.10 0.15 0.7524 6000 0.10 0.15 0.75
25 6000 0.10 0.30 0.60 实施例 26〜28 25 6000 0.10 0.30 0.60 Example 26~28
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )和 3份 0.0133g ( 0.0002065mol )实施例 2制备的羧基化的阿霉素衍生物，放入 3个反应 瓶中， 再分别加入 0.02284g、 0.06853g和 0.1371g 的叶酸， 0.49488g、 0.69282g 和 0.98975g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.03154g、 0.04415g和 0.06307g的 4-二曱氨基吡啶， 混合均匀后， 再分 别加入 25mL二曱基亚砜溶解反应体系，室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton的透 析袋透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉 素键合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) and 3 parts of 0.01133 g (0.0002065 mol) of the carboxylated doxorubicin derivative prepared in Example 2 were placed in 3 reaction bottles, and then separately added. 0.02284 g, 0.06853 g and 0.1371 g of folic acid, 0.49488 g, 0.69282 g and 0.98975 g of 1-(3-diamidinopropyl)-3-ethylcarbodiimide' hydrochloride, 0.03154 g, 0.04415 g After mixing with 0.06307 g of 4-diaminopyridine, the reaction system was dissolved in 25 mL of dimercaptosulfoxide, and reacted at room temperature for 3 days under light agitation. After the reaction was completed, the obtained reaction mixture was respectively used. The dialysis bag with a molecular weight cut off of 3500 Dalton was dialyzed for 3 days, and the water was changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 26〜28 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 j、 k取 值见表 8。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 26 to 28 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, j and k are shown in Table 8. .
表 8 本发明实施例 26〜28制备的高分子阿霉素键合药 h、 j、 k取值 实施例 Mn (g ηιοΓ1) h j k Table 8 Examples of the polymer doxorubicin bonding drugs h, j, k prepared in Examples 26 to 28 of the present invention. Example M n (g ηιοΓ 1 ) hjk
26 6000 0.20 0.05 0.75 26 6000 0.20 0.05 0.75
27 6000 0.20 0.15 0.6527 6000 0.20 0.15 0.65
28 6000 0.20 0.30 0.50 实施例 29〜31 28 6000 0.20 0.30 0.50 Example 29~31
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )和 3份 0.20001g (0.0003098mol)实施例 2制备的羧基化的阿霉素衍生物， 放入 3个反应瓶 中，再分别加入 0.02284g、 0.06853g和 0.1371g的叶酸， 0.69294g、 0.89092g 和 1.18790g的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.04416g、 0.05677g和 0.07570g的 4-二曱氨基吡啶， 混合均匀后,再分别加入 25mL 二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton 的透析袋透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键合药。 3 parts of 0.16768 g of dextran (number average molecular weight of 6000) and 3 parts of 0.20001 g (0.0003098 mol) of the carboxylated doxorubicin derivative prepared in Example 2, placed in three reaction flasks, and then added 0.02284 g, 0.06853 g, and 0.1371 g of folic acid, 0.69294 g, 0.89092 g, and 1.18790 g, respectively. -(3-Diaminoaminopropyl)-3-ethylcarbodiimide' hydrochloride, 0.04416g, 0.05677g and 0.07570g of 4-diaminopyridine, after mixing uniformly, add 25mL of diterpene separately The sulfoxide is dissolved in the reaction system, and reacted at room temperature for 3 days under light and stirring conditions. After the reaction is completed, the obtained reaction mixture is dialyzed against a dialysis bag having a molecular weight cutoff of 3,500 Dalton for 3 days, and the water is changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 29〜31 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 j、 k取 值见表 9。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 29 to 31 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, j and k are shown in Table 9. .
表 9本发明实施例 29〜31制备的高分子阿霉素键合药 h、 j、 k取值 实施例 Mn (g ηιοΓ1) h j k Table 9 Examples of the polymer doxorubicin bonding drugs h, j, k prepared in Examples 29 to 31 of the present invention. Example M n (g ηιοΓ 1 ) hjk
29 6000 0.30 0.05 0.65 29 6000 0.30 0.05 0.65
30 6000 0.30 0.15 0.5530 6000 0.30 0.15 0.55
31 6000 0.30 0.30 0.40 实施例 32〜35 31 6000 0.30 0.30 0.40 Example 32~35
将 4份 0.16768g的葡聚糖（数均分子量为 6000 )，放入 4个反应瓶中， 分别加入 0.01447g ( 0.0000207mol )、 0.07233g ( 0.0001034mol )、 0.14465g ( 0.0002067mol )和 0.21698g(0.0003101mol) 实施例 4制备的^^化的阿 霉素衍生物， 0.03968g、 0.19824g、 0.39629g和 0.59452g的 1-(3-二曱氨基 丙基) -3-乙基碳二亚胺 '盐酸盐， 0.00253g、 0.01263g、 0.02525g和 0.03788g 的 4-二曱氨基吡啶， 混合均匀后， 再分别加入 25mL二曱基亚砜溶解反 应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反 应混合液用截留分子量为 3500 Dalton的透析袋透析 3 天， 每隔 5 h换水 一次， 将反应液冻干， 分别得到高分子阿霉素键合药。  4 parts of 0.16768 g of dextran (number average molecular weight of 6000) were placed in 4 reaction flasks, and 0.01447 g (0.0000207 mol), 0.07233 g (0.0001034 mol), 0.14465 g (0.0002067 mol) and 0.21698 g, respectively, were added. (0.0003101 mol) of the doxorubicin derivative prepared in Example 4, 0.03968 g, 0.19824 g, 0.39629 g, and 0.59452 g of 1-(3-diaminoaminopropyl)-3-ethylcarbazone Amine's hydrochloride, 0.00253g, 0.01263g, 0.02525g and 0.03788g of 4-diaminopyridine, after mixing uniformly, add 25mL of dimercaptosulfoxide to dissolve the reaction system, react at room temperature and avoid light. After the reaction, the reaction mixture was dialyzed against a dialysis bag with a molecular weight cutoff of 3,500 Dalton for 3 days, and the water was changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond. .
分别对本发明实施例 32〜35 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 k取值 见表 10。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 32 to 35 of the present invention were respectively carried out, and the results showed that they all have the structure of the formula (I), wherein the values of h and k are shown in Table 10.
表 10本发明实施例 30〜32制备得到的高分子阿霉素键合药 h、 k取 实施例 Mn (g mol"1) h k Table 10 The polymer doxorubicin bonding drugs h, k prepared in the examples 30 to 32 of the present invention were taken. Example M n (g mol" 1 ) hk
32 6000 0.02 0.98 32 6000 0.02 0.98
33 6000 0.10 0.9033 6000 0.10 0.90
34 6000 0.20 0.8034 6000 0.20 0.80
35 6000 0.30 0.70 实施例 36-38 35 6000 0.30 0.70 Examples 36-38
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )，放入 3个反应瓶中， 分别加入 0.01447g ( 0.0000207mol )实施例 4制备的羧基化的阿霉素衍生 物， 0.01854g、 0.05563 和 0.11125g 的乳糖酸， 0.13890g、 0.33733g和 0.6350g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 .盐酸盐， 0.00885g、 0.02150g和 0.04046g的 4-二曱氨基吡啶， 混合均匀后,再分别加入 25mL 二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton 的透析袋透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) were placed in three reaction flasks, and 0.01447 g (0.0000 207 mol) of the carboxylated doxorubicin derivatives prepared in Example 4, 0.01854 g, were respectively added. 0.05563 and 0.11125g of lactobionic acid, 0.13890g, 0.33733g and 0.6350g of 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide hydrochloride, 0.00885g, 0.02150g and 0.04046g After the mixture is uniformly mixed, 25 mL of dimercaptosulfoxide is added to dissolve the reaction system, and the reaction is carried out at room temperature for 3 days under light agitation. After the reaction is completed, the obtained reaction mixture is respectively subjected to molecular weight cut off. The 3500 Dalton dialysis bag was dialyzed for 3 days, the water was changed every 5 hours, and the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 36〜38 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取 值见表 11。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 36 to 38 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 11. .
表 11本发明实施例 36〜38制备得到的高分子阿霉素键合药 h、 i、 k 取值  Table 11 In the present invention, the polymer doxorubicin bonding drugs prepared by the preparation of the compounds 36 to 38 have values of h, i, k
实施例 Mn (g mol"1) h i k Example M n (g mol" 1 ) hik
36 6000 0.02 0.05 0.93 36 6000 0.02 0.05 0.93
37 6000 0.02 0.15 0.8337 6000 0.02 0.15 0.83
38 6000 0.02 0.30 0.68 实施例 39〜41 38 6000 0.02 0.30 0.68 Example 39~41
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )，放入 3个反应瓶中， 分别加入 0.07233g ( 0.0001034mol )实施例 4制备的羧基化的阿霉素衍生 物， 0.01854g、 0.05563 和 0.11125g 的乳糖酸， 0.29735g、 0.49560g和 0.79295g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 .盐酸盐， 0.01895g、 0.03158g和 0.05053g的 4-二曱氨基吡啶， 混合均匀后,再分别加入 25mL 二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton 的透析袋透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键合药。 3 parts of 0.16768 g of dextran (number average molecular weight of 6000) were placed in three reaction flasks, and 0.07233 g (0.0001034 mol) of the carboxylated doxorubicin derivatives prepared in Example 4, 0.01854 g, were respectively added. 0.05563 and 0.11125g of lactobionic acid, 0.29735g, 0.49560g and 0.79295g of 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide hydrochloride, 0.01895g, 0.03158g and 0.05053g of 4-diaminopyridine, after mixing uniformly, add 25mL of dimercaptosulfoxide to dissolve the reaction system, react at room temperature for 3 days under light agitation, and after the reaction is finished, separately mix the obtained reaction. The solution was dialyzed against a dialysis bag having a molecular weight cutoff of 3,500 Dalton for 3 days, and the water was changed every 5 hours, and the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 39〜41 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取 值见表 12。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 39 to 41 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 12. .
表 12本发明实施例 39〜41制备得到的高分子阿霉素键合药 h、 i、 k 取值  Table 12 Inventive Examples 39 to 41 prepared polymer doxorubicin bonding drugs h, i, k
实施例 Mn (g mol"1) h i k Example M n (g mol" 1 ) hik
39 6000 0.10 0.05 0.85 39 6000 0.10 0.05 0.85
40 6000 0.10 0.15 0.7540 6000 0.10 0.15 0.75
41 6000 0.10 0.30 0.60 实施例 42〜44 41 6000 0.10 0.30 0.60 Example 42~44
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )，放入 3个反应瓶中， 分别加入 0.14465g ( 0.0002067mol )实施例 4制备的羧基化的阿霉素衍生 物； 0.01854g、 0.05563 和 0.11125g 的乳糖酸， 0.49536g、 0.69350g和 0.98787g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.03157g、 0.04419g和 0.06313 lg的 4-二曱氨基吡啶，混合均匀后，再分别加入 25mL 二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton 的透析袋透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) were placed in three reaction flasks, and 0.14465 g (0.0002067 mol) of the carboxylated doxorubicin derivatives prepared in Example 4 were separately added; 0.01854 g, 0.05563 and 0.11125g of lactobionic acid, 0.49536g, 0.69350g and 0.98787g of 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide' hydrochloride, 0.03157g, 0.04419g and 0.06313 lg After the mixture is uniformly mixed, 25 mL of dimercaptosulfoxide is added to dissolve the reaction system, and the reaction is carried out at room temperature for 3 days under light agitation. After the reaction is completed, the obtained reaction mixture is respectively subjected to molecular weight cut off. The 3500 Dalton dialysis bag was dialyzed for 3 days, the water was changed every 5 hours, and the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 42〜44制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取 值见表 13。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 42 to 44 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 13. .
表 13本发明实施例 42〜44制备得到的高分子阿霉素键合药 h、 i、 k 取值  Table 13 Inventive Example 42 to 44 prepared polymer doxorubicin bonding drugs h, i, k values
实施例 Mn (g mol"1) h i k Example M n (g mol" 1 ) hik
42 6000 0.20 0.05 0.75 43 6000 0.20 0.15 0.65 44 6000 O20 030 0.50 实施例 45-47 42 6000 0.20 0.05 0.75 43 6000 0.20 0.15 0.65 44 6000 O20 030 0.50 Example 45-47
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )，放入 3个反应瓶中， 分别加入 0.21698g(0.0003101mol)实施例 4 制备的羧基化的阿霉素衍生 物， 0.01854g、 0.05563 和 0.11125g 的乳糖酸， 0.69361g、 0.89178g和 1.18904g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.044199g、 0.05683g和 0.075770g的 4-二曱氨基吡啶，混合均勾后，再分别加入 25mL 二曱基亚砜溶解反应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton 的透析袋透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) were placed in three reaction flasks, and 0.21698 g (0.0003101 mol) of the carboxylated doxorubicin derivatives prepared in Example 4, 0.01854 g, were respectively added. 0.05563 and 0.11125g of lactobionic acid, 0.69361g, 0.89178g and 1.18904g of 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide' hydrochloride, 0.044199g, 0.05683g and 0.075770g 4-diaminopyridine, after mixing, add 25mL of dimercaptosulfoxide to dissolve the reaction system, react at room temperature for 3 days under light agitation, and after the reaction, separate the obtained reaction mixture with molecular weight cut-off The plate was dialyzed for 3 days in a 3500 Dalton dialysis bag, and the water was changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 45〜47 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取 值见表 14。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 45 to 47 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 14. .
表 14本发明实施例 45〜47制备得到的高分子阿霉素键合药 h、 i、 k 取值  Table 14 Inventive Example 45~47 Preparation of polymer doxorubicin bonding drugs h, i, k
实施例 Mn (g ηιοΓ1) h i k Example M n (g ηιοΓ 1 ) hik
45 6000 0.30 0.05 0.65 45 6000 0.30 0.05 0.65
46 6000 0.30 0.15 0.5546 6000 0.30 0.15 0.55
47 6000 0.30 0.30 0.40 实施例 48〜51 47 6000 0.30 0.30 0.40 Example 48~51
将 4份 0.16768g葡聚糖 (数均分子量为 6000 ), 放入 4个反应瓶中， 分别加入 0.00133g ( 0.0000207mol )、 0.06667g ( 0.00010327mol )、 0.13333g ( 0.0002065mol )和 0.20001g(0.0003098mol) 实施例 2制备的羧基化的阿 霉素衍生物， 0.03968g、 0.19797g、 0.39586g和 0.59235g的 1-(3-二曱氨基 丙基) -3-乙基碳二亚胺 '盐酸盐， 0.00253g、 0.01262g、 0.02523g和 0.03785g 的 4-二曱氨基吡啶， 混合均匀后， 再分别加入 25mL二曱基亚砜溶解反 应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反 应混合液用截留分子量为 3500 Dalton的透析袋透析 3 天， 每隔 5 h换水 一次， 将反应液冻干， 分别得到高分子阿霉素键合药。 4 parts of 0.16768 g of dextran (number average molecular weight of 6000) were placed in 4 reaction flasks, and 0.00133 g (0.0000207 mol), 0.06667 g (0.00010327 mol), 0.13333 g (0.0002065 mol) and 0.20001 g ( 0.0003098 mol) The carboxylated doxorubicin derivative prepared in Example 2, 0.03968 g, 0.19797 g, 0.39586 g, and 0.59235 g of 1-(3-diamidinopropyl)-3-ethylcarbodiimide' Hydrochloride, 0.00253g, 0.01262g, 0.02523g and 0.03785g of 4-diaminopyridine, after mixing uniformly, add 25mL of dimercaptosulfoxide to dissolve the reaction system, and react at room temperature for 3 days under light and stirring conditions. After the reaction, the obtained reaction mixture was dialyzed against a dialysis bag having a molecular weight cutoff of 3,500 Dalton for 3 days, and the water was changed every 5 hours. Once, the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 48〜51 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 k取值 见表 1。  The polymer magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 48 to 51 of the present invention were respectively carried out, and the results showed that they all have the structure of the formula (I), wherein the values of h and k are shown in Table 1.
表 15 本发明实施例 48〜51制 到的高分子阿霉素键合药 h、 k取
Figure imgf000025_0001
Table 15 The polymer doxorubicin bonding drugs h, k prepared in Examples 48 to 51 of the present invention were taken.
Figure imgf000025_0001
实施例 Mn (g mol"1) h k Example M n (g mol" 1 ) hk
48 6000 0.02 0.98 48 6000 0.02 0.98
49 6000 0.10 0.9049 6000 0.10 0.90
50 6000 0.20 0.8050 6000 0.20 0.80
51 6000 0.30 0.70 实施例 52〜54 51 6000 0.30 0.70 Example 52~54
将 3份葡聚糖 0.16768g (数均分子量为 6000 ), 放入 3个反应瓶中， 分别加入 0.07233g ( 0.0001034mol )实施例 4制备的羧基化的阿霉素衍生 物， 0.02284g、 0.06853和 0.1371g的叶酸； 0.29735g、 0.49560g和 0.79295g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.01895g、 0.03158g 和 0.05053g的 4-二曱氨基吡啶， 混合均匀后， 再分别加入 25mL二曱基亚 砜溶解反应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将 得到的反应混合液用截留分子量为 3500 Dalton的透析袋透析 3 天，每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键合药。  3 parts of dextran 0.16768g (number average molecular weight of 6000) was placed in three reaction flasks, and 0.07233g (0.0001034mol) of the carboxylated doxorubicin derivatives prepared in Example 4 were added, respectively, 0.02284g, 0.06853 And 0.1371 g of folic acid; 0.29735 g, 0.49560 g and 0.79295 g of 1-(3-diamidinopropyl)-3-ethylcarbodiimide' hydrochloride, 0.01895 g, 0.03158 g and 0.05053 g of 4 - Di-aminopyridine, after mixing uniformly, adding 25 mL of dimethylsulfoxide to dissolve the reaction system, reacting at room temperature for 3 days under light agitation, and after completion of the reaction, respectively, the obtained reaction mixture has a molecular weight cutoff of 3,500 Dalton. The dialysis bag was dialyzed for 3 days, and the water was changed every 5 hours, and the reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 52〜54制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 j、 k取 值见表 16。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 52 to 54 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, j and k are shown in Table 16. .
表 16本发明实施例 52〜54制备得到的高分子阿霉素键合药 h、 j、 k 取值  Table 16 In the present invention, the polymer doxorubicin bonding drugs prepared by the preparation of the formulas 52 to 54 h, j, k
实施例 Mn (g mol"1) h j k Example M n (g mol" 1 ) hjk
52 6000 0.10 0.05 0.85 52 6000 0.10 0.05 0.85
53 6000 0.10 0.15 0.7553 6000 0.10 0.15 0.75
54 6000 0.10 0.30 0.60 实施例 55〜57 54 6000 0.10 0.30 0.60 Examples 55 to 57
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )，放入 3个反应瓶中， 分别加入 0.14465g ( 0.0002067mol )实施例 4制备的羧基化的阿霉素衍生 物， 0.02281g、0.06843和 0.13686g的叶酸，0.49536§、0.6935(^和0.98787§ 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.03157g、 0.04419g 和 0.06313 lg的 4-二曱氨基吡啶， 混合均匀后， 再分别加入 25mL二曱基亚 砜溶解反应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将 得到的反应混合液用截留分子量为 3500 Dalton的透析袋透析 3 天，每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) were placed in three reaction flasks, and 0.14465 g (0.0002067 mol) of the carboxylated doxorubicin derivative prepared in Example 4, 0.02281 g, was added, respectively. 0.06843 and 0.13686 g of folic acid, 0.49536 §, 0.6935 (^ and 0.98787§ 1-(3-diaminopropyl)-3-ethylcarbodiimide' hydrochloride, 0.03157 g, 0.04419 g and 0.06313 lg After the mixture is uniformly mixed, 25 mL of dimethyl sulfoxide is added to dissolve the reaction system, and the reaction is carried out at room temperature for 3 days under light agitation. After the reaction is completed, the obtained reaction mixture is respectively subjected to a molecular weight cut off. The 3500 Dalton dialysis bag was dialyzed for 3 days, and the water was changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 55〜57 制备得到的高分子阿霉素键合药进行核 磁共振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 j、 k取值见表 17。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 55 to 57 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, j and k are shown in the table. 17.
表 17本发明实施例 55〜57制备得到的高分子阿霉素键合药 h、 j、 k 取值  Table 17 Inventive Examples 55 to 57 prepared polymer doxorubicin bonding drugs h, j, k values
实施例 Mn (g mol"1) h j k Example M n (g mol" 1 ) hjk
55 6000 0.20 0.05 0.75 55 6000 0.20 0.05 0.75
56 6000 0.20 0.15 0.6556 6000 0.20 0.15 0.65
57 6000 0.20 0.30 0.50 实施例 58〜60 57 6000 0.20 0.30 0.50 Example 58~60
将 3份 0.16768g的葡聚糖（数均分子量为 6000 )，放入 3个反应瓶中， 分别加入 0.21698g(0.0003101mol)实施例 4 制备的羧基化的阿霉素衍生 物， 0.02281g、0.06839和 0.13688g的叶酸，0.69361g、0.89178g和 1.18904g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.044199g、 0.05683g和 0.075770g的 4-二曱氨基吡啶， 混合均匀后, 再分别加入 25mL二曱基亚 砜溶解反应体系， 室温避光搅拌条件下反应 3 天， 反应结束后， 分别将 得到的反应混合液用截留分子量为 3500 Dalton的透析袋透析 3 天，每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉素键合药。  3 parts of 0.16768 g of dextran (number average molecular weight of 6000) were placed in three reaction flasks, and 0.21698 g (0.0003101 mol) of the carboxylated doxorubicin derivatives prepared in Example 4, 0.02281 g, were respectively added. 0.06839 and 0.13688g of folic acid, 0.69361g, 0.89178g and 1.18904g of 1-(3-diaminoaminopropyl)-3-ethylcarbodiimide' hydrochloride, 0.044199g, 0.05683g and 0.075770g After the mixture was uniformly mixed, 25 mL of dimethylsulfoxide was added to dissolve the reaction system, and the reaction was carried out at room temperature for 3 days under light agitation. After the reaction was completed, the obtained reaction mixture was respectively subjected to a molecular weight cutoff of 3,500. Dalton's dialysis bag was dialyzed for 3 days, and water was changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 58〜60制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 j、 k取 值见表 18。 The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 58 to 60 of the present invention respectively showed that they all have the structure of the formula (I), wherein h, j, k take The values are shown in Table 18.
表 18本发明实施例 58〜60制备得到的高分子阿霉素键合药 h、 j、 k 取值  Table 18 Examples of the polymer doxorubicin bonding drugs prepared by the methods 58 to 60, h, j, k
实施例 Mn (g mol"1) h j k Example M n (g mol" 1 ) hjk
58 6000 0.30 0.05 0.65 58 6000 0.30 0.05 0.65
59 6000 0.30 0.15 0.5559 6000 0.30 0.15 0.55
60 6000 0.30 0.30 0.40 实施例 61〜63 60 6000 0.30 0.30 0.40 Example 61~63
将 3份 0.16768g的葡聚糖 (数均分子量为 40000 )和 3份 0.00133g 3 parts of 0.16768g dextran (number average molecular weight of 40,000) and 3 parts of 0.00133g
( 0.0000207mol )实施例 2制备的羧基化的阿霉素衍生物，放入 3个反应 瓶中， 再分别加入 0.01854g、 0.05563g和 0.11125g的乳糖酸， 0.13890g、 0.33731g 和 0.63325g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.00885g、 0.02150g和 0.04046g的 4-二曱氨基吡啶， 混合均匀后, 再分 别加入 25mL二曱基亚砜溶解反应体系，室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton的透 析袋透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉 素键合药。 (0.0000207 mol) The carboxylated doxorubicin derivative prepared in Example 2 was placed in three reaction flasks, and 0.01854 g, 0.05563 g, and 0.11125 g of lactobionic acid, 0.13890 g, 0.33731 g, and 0.63325 g, respectively, were added. 1-(3-Diaminoaminopropyl)-3-ethylcarbodiimide' hydrochloride, 0.00885g, 0.02150g and 0.04046g of 4-diaminopyridine, after mixing, add 25mL two The mercapto sulfoxide was dissolved in the reaction system, and reacted at room temperature for 3 days under light agitation. After the reaction was completed, the obtained reaction mixture was dialyzed against a dialysis bag having a molecular weight cutoff of 3,500 Dalton for 3 days, and the water was changed every 5 hours. The reaction solution was lyophilized to obtain a polymer doxorubicin bond.
分别对本发明实施例 61〜63 制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取 值见表 19。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 61 to 63 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 19. .
表 19本发明实施例 61〜63制备得到的高分子阿霉素键合药 h、 i、 k 取值  Table 19 Inventive Example 61 to 63 prepared polymer doxorubicin bonding drugs h, i, k values
实施例 Mn (g mol"1) h i k Example M n (g mol" 1 ) hik
61 40000 0.02 0.05 0.93 61 40000 0.02 0.05 0.93
62 40000 0.02 0.15 0.8362 40000 0.02 0.15 0.83
63 40000 0.02 0.30 0.68 实施例 64〜66 63 40000 0.02 0.30 0.68 Example 64~66
将 3份 0.16768g的葡聚糖（数均分子量为 100000 )和 3份 0.00133g ( 0.0000207mol )实施例 2制备的羧基化的阿霉素衍生物，放入 3个反应 瓶中， 再分别加入 0.01854g、 0.05563g和 0.11125g的乳糖酸， 0.13890g、 0.33731g 和 0.63325g 的 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐， 0.00885g、 0.02150g和 0.04046g的 4-二曱氨基吡啶， 混合均匀后, 再分 别加入 25mL二曱基亚砜溶解反应体系，室温避光搅拌条件下反应 3 天， 反应结束后， 分别将得到的反应混合液用截留分子量为 3500 Dalton的透 析袋透析 3 天， 每隔 5 h换水一次， 将反应液冻干， 分别得到高分子阿霉 素键合药。 3 parts of 0.16768 g of dextran (number average molecular weight of 100,000) and 3 parts of 0.00133 g (0.0000207 mol) of the carboxylated doxorubicin derivative prepared in Example 2 were placed in 3 reactions. In the bottle, 0.01854g, 0.05563g and 0.11125g of lactobionic acid, 0.13890g, 0.33731g and 0.63325g of 1-(3-diaminopropyl)-3-ethylcarbodiimide hydrochloride were added. Salt, 0.00885g, 0.02150g and 0.04046g of 4-diaminopyridine, after mixing uniformly, add 25mL of dimethyl sulfoxide to dissolve the reaction system, react at room temperature for 3 days under light agitation, after the reaction, respectively The obtained reaction mixture was dialyzed against a dialysis bag having a molecular weight cut off of 3,500 Dalton for 3 days, and water was exchanged every 5 hours, and the reaction solution was lyophilized to obtain a polymer doxorubicin-bonded drug.
分别对本发明实施例 64〜66制备的高分子阿霉素键合药进行核磁共 振分析和红外分析， 结果表明， 其均具有式（I )结构， 其中， h、 i、 k取 值见表 20。  The nuclear magnetic resonance analysis and infrared analysis of the polymer doxorubicin-bonded materials prepared in Examples 64 to 66 of the present invention respectively showed that they all have the structure of the formula (I), wherein the values of h, i and k are shown in Table 20. .
表 20本发明实施例 64〜66制备得到的高分子阿霉素键合药 h、 i、 k 取值  Table 20 Inventive Example 64 to 66 prepared polymer doxorubicin bonding drugs h, i, k values
实施例 Mn (g mol"1) h i k Example M n (g mol" 1 ) hik
64 100000 0.02 0.05 0.93  64 100000 0.02 0.05 0.93
65 100000 0.02 0.15 0.83  65 100000 0.02 0.15 0.83
66 100000 0.02 0.30 0.68 以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的 普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进 和润饰， 这些改进和润饰也应视为本发明的保护范围。  66 100000 0.02 0.30 0.68 The above is only a preferred embodiment of the present invention, and it should be noted that those skilled in the art can make several improvements and modifications without departing from the principles of the present invention. These improvements and finishes should also be considered as protection of the present invention.
-I- -I-

Claims

权 利 要 求 Rights request
1、 阿霉素键合药， 包括具有式（I) 结构的第一重复单元、 式（II) 结构的第二重复单元、 式（III) 结构的第三重复单元和式（IV) 结构的 第四重复单元： 1. Doxorubicin bonded drug, including a first repeating unit having a structure of formula (I), a second repeating unit of a structure of formula (II), a third repeating unit of a structure of formula (III) and a structure of formula (IV). Fourth repeating unit:
（I); (I);
式 (II) Formula (II)
（ III )； (III);
Figure imgf000029_0001
（IV);
Figure imgf000029_0001
(IV);
R选自式（ 1 )〜（ 4 )结构中的一种: R is selected from one of the structures of formulas (1) to (4):
0 OH 0 0 OH 0
Figure imgf000029_0002
Figure imgf000030_0001
Figure imgf000029_0002
Figure imgf000030_0001
Lactonibicacid为乳糖酸， 其具有式 (5) 结构: Lactonibicacid is lactobionic acid, which has the structure of formula (5):
Figure imgf000030_0002
Figure imgf000030_0002
(5); (5);
FA为叶酸， 其具有式（6)结构: FA is folic acid, which has the structure of formula (6):
H2N
Figure imgf000030_0003
(6); H 2 N
Figure imgf000030_0003
(6);
式（I)中， 所述第一重复单元的摩尔数 h、 第二重复单元的摩尔数 第三重复单元的摩尔数^ 第四重复单元的摩尔数 k与第一重复单元、 第 二重复单元、第三重复单元和第四重复单元的总摩尔数的比例满足以下条 件： 0.02<h/(h+i+j+k)<0.3; 0<i/(h+i+j+k)<0.3; 0≤j/(h+i+j+k)≤0.3。 In formula (I), the number of moles of the first repeating unit h, the number of moles of the second repeating unit h, the number of moles of the third repeating unit h, the number of moles k of the fourth repeating unit, and the number of moles k of the first repeating unit and the second repeating unit. , the ratio of the total number of moles of the third repeating unit and the fourth repeating unit satisfies the following conditions: 0.02<h/(h+i+j+k)<0.3; 0<i/(h+i+j+k)< 0.3; 0≤j/(h+i+j+k)≤0.3.
2、 根据权利要求 1 所述的阿霉素键合药， 其特征在于， 2. The doxorubicin bonded drug according to claim 1, characterized in that,
0.15<h/(h+i+j+k)<0.3 , 0.10<i/(h+i+j+k)<0.20, 0.10<j/(h+i+j+k)<0.20„0.15<h/(h+i+j+k)<0.3, 0.10<i/(h+i+j+k)<0.20, 0.10<j/(h+i+j+k)<0.20„
3、 根据权利要求 1所述的阿霉素键合药， 其特征在于， 所述阿霉素 键合药的数均分子量为 6000〜100000。 3. The doxorubicin-bonded drug according to claim 1, wherein the number average molecular weight of the doxorubicin-bonded drug is 6,000 to 100,000.
4、 阿霉素键合药的制备方法， 包括以下步骤： 4. The preparation method of doxorubicin bonded drug includes the following steps:
葡聚糖、 羧基化阿霉素衍生物、 具有式（V)结构的乳糖酸和具有式 (VI)结构的叶酸与 1-(3-二曱氨基丙基 )-3-乙基碳二亚胺 '盐酸盐和 4-二 曱氨基吡啶在有机溶剂中反应，得到阿霉素键合药，所述羧基化阿霉素衍 生物具有式（VII-a)、 式 (VII-b)、 式 ( VII-c )或式 ( VII-d )结构： Dextran, carboxylated doxorubicin derivatives, lactobionic acid with a structure of formula (V) and folic acid with a structure of formula (VI) and 1-(3-dimethylaminopropyl)-3-ethylcarbodioxide Amine hydrochloride and 4-dimethylaminopyridine react in an organic solvent to obtain a doxorubicin bonded drug. The carboxylated doxorubicin derivative has formula (VII-a), formula (VII-b), Structure of formula (VII-c) or formula (VII-d):
Figure imgf000031_0001
Figure imgf000031_0001
(V); (V);
Figure imgf000031_0002
Figure imgf000031_0002
(VI) (VI)
Figure imgf000032_0001
Figure imgf000032_0001
5、 根据权利要求 4所述的制备方法， 其特征在于， 所述羧基化阿霉 素衍生物、 乳糖酸、 叶酸与葡聚糖主链上葡萄糖重复单元的摩尔比为 (2〜30) ： (0〜30)： (0〜30)： 謂。 5. The preparation method according to claim 4, characterized in that the molar ratio of the carboxylated doxorubicin derivative, lactobionic acid, folic acid and glucose repeating units on the dextran main chain is (2~30): (0~30): (0~30): Predicate.
6、 根据权利要求 4所述的制备方法， 其特征在于， 所述羧基化阿霉 素衍生物、 乳糖酸、 叶酸与葡聚糖主链上葡萄糖重复单元的摩尔比为 6. The preparation method according to claim 4, characterized in that the molar ratio of the carboxylated doxorubicin derivative, lactobionic acid, folic acid and glucose repeating units on the dextran main chain is
(8〜25) ： (5〜25)： (5〜25)： 謂。 (8~25): (5~25): (5~25): Predicate.
7、 根据权利要求 4所述的制备方法， 其特征在于， 所述葡聚糖的分 子量为 6000〜100000。 7. The preparation method according to claim 4, characterized in that the molecular weight of the dextran is 6000~100000.
8、 根据权利要求 4所述的制备方法， 其特征在于， 所述有机溶剂为 二曱基亚砜。 8. The preparation method according to claim 4, characterized in that the organic solvent is dimethyl sulfoxide.
9、 根据权利要求 4所述的制备方法， 其特征在于， 所述羧基化阿霉 素衍生物按照以下方法制备： 9. The preparation method according to claim 4, characterized in that the carboxylated doxorubicin derivative is prepared according to the following method:
阿霉素 .盐酸盐、 酸酐和三乙胺在有机溶剂中反应， 得到羧基化的阿 霉素衍生物； 所述酸酐选自 1,2-二羧基环己烯酸酐、 丁二酸酐、 2,2,3,3- 四曱基丁二酸酐和顺 -3-羧基戊烯二酸酐中的一种。 Doxorubicin. Hydrochloride, acid anhydride and triethylamine react in an organic solvent to obtain a carboxylated doxorubicin derivative; the acid anhydride is selected from 1,2-dicarboxycyclohexene anhydride, succinic anhydride, 2 , one of 2,3,3-tetramethylsuccinic anhydride and cis-3-carboxyglutenic anhydride.
10、 根据权利要求 6所述的制备方法， 其特征在于， 所述阿霉素-盐 酸盐、 酸酐和三乙胺的摩尔比为 1: (0.5〜2) ： (0.5〜2)。 10. The preparation method according to claim 6, characterized in that, the doxorubicin-salt The molar ratio of acid salt, acid anhydride and triethylamine is 1: (0.5~2): (0.5~2).
+ +
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