WO2017128963A1 - 一种化合物及其应用以及一种铂类配合物及其脂质体 - Google Patents

一种化合物及其应用以及一种铂类配合物及其脂质体 Download PDF

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WO2017128963A1
WO2017128963A1 PCT/CN2017/071114 CN2017071114W WO2017128963A1 WO 2017128963 A1 WO2017128963 A1 WO 2017128963A1 CN 2017071114 W CN2017071114 W CN 2017071114W WO 2017128963 A1 WO2017128963 A1 WO 2017128963A1
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platinum complex
cooh
platinum
liposome
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杨丽
曾群
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Beijing Snowle Bio-Tech Co Ltd
Shenyang Pharmaceutical University
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Beijing Snowle Bio-Tech Co Ltd
Shenyang Pharmaceutical University
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Priority to US16/072,614 priority Critical patent/US10730898B2/en
Priority to JP2018557176A priority patent/JP6744429B2/ja
Priority to EP17743578.1A priority patent/EP3409682B1/en
Publication of WO2017128963A1 publication Critical patent/WO2017128963A1/zh
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/28—Compounds containing heavy metals
    • A61K31/282—Platinum compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/74—Synthetic polymeric materials
    • A61K31/80—Polymers containing hetero atoms not provided for in groups A61K31/755 - A61K31/795
    • 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
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/45—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • C07C233/46—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
    • C07C233/47—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to a hydrogen atom or to a carbon atom of an acyclic saturated carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F15/00—Compounds containing elements of Groups 8, 9, 10 or 18 of the Periodic Table
    • C07F15/0006—Compounds containing elements of Groups 8, 9, 10 or 18 of the Periodic Table compounds of the platinum group
    • C07F15/0086—Platinum compounds
    • C07F15/0093—Platinum compounds without a metal-carbon linkage
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
    • C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
    • C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates

Definitions

  • the present invention relates to the field of medicine, and in particular to a compound and its use, and a platinum complex and a liposome thereof.
  • platinum anti-tumor drugs Since the American scientist B. Rosenborg accidentally discovered that cisplatin has anti-tumor activity in 1965, the synthetic application and research of platinum-based metal anticancer drugs have developed rapidly, compared with traditional cytotoxic antitumor drugs, platinum anti-tumor drugs.
  • the mechanism of action of oncology drugs is unique. Its main target is DNA.
  • platinum antitumor drugs enter the cell They are hydrolyzed to form positively charged hydrated platinum with high activity. Hydrated platinum reacts with nucleic acids and proteins to form interchain conjugates in DNA strands. , which leads to changes in the secondary structure of DNA, hinders transcription and replication of DNA, and ultimately leads to apoptosis.
  • platinum ie, cisplatin, cis-1,1-cyclobutanedicarboxylic acid diamine platinum, carboplatin, cis-oxalic acid-(reverse)
  • Platinum of the formula -(-)-1,2-cyclohexanediamine) is Oxaliplatin, Nedaplatin, Eptaplatin, Lobaplatin and the like.
  • Platinum anti-tumor drugs have the characteristics of wide anti-cancer spectrum, strong activity and no cross-resistance. Statistics show that 70% to 80% of all chemotherapy regimens are platinum-based or platinum-based drugs. However, platinum-based anticancer drugs lack specificity for tumor cells.
  • the toxic side effects It mainly includes nephrotoxicity, gastrointestinal toxicity (nausea, vomiting), bone marrow toxicity, neurotoxicity and ototoxicity, etc., and the concentration of the drug reaching the tumor site is low, which reduces the therapeutic effect, and the long-term application may also cause cancer cells to develop drug resistance. , thereby reducing the shortcomings of its chemotherapy.
  • the prior art discloses a method of developing a platinum-based formulation in a manner in which a plurality of carriers carry a drug.
  • Uchino et al. used a combination of cisplatin and carboxyl-rich polyethylene glycol-b-polyglutamic acid to prepare a micelle with a drug loading of 30 wt.% (NC-6004) has entered Phase II clinical studies [British] Journal of Cancer (2005) 93, 678-687], however, since the obtained micelles are formed by cross-linking between polyamino acid side chains, the lyophilized powder obtained by this method may be difficult to reconstitute after lyophilization. And the efficacy of the preparation is not improved.
  • Access Pharmaceuticals combines the commonly used polymer HPMA with cisplatin to prepare AP5280.
  • the tumor aggregation of AP5280 is 19 times that of cisplatin injection and significantly reduces the nephrotoxicity of cisplatin [J. Drug Target 10 (2002) 549 -556].
  • These studies have shown that platinum compounds can bind to macromolecules through environmentally sensitive chemical bonds, thereby effectively reducing the toxicity of platinum drugs and improving efficacy.
  • different environmentally sensitive chemical bonds are essential for the efficacy of platinum anticancer drugs because they can change the hydration rate of platinum compounds in the body. The faster the hydration rate of platinum compounds in the body, the stronger the antitumor effect. But at the same time, its toxicity is also greater.
  • Poulomi Sengupta et al. combined the effect of cisplatin with cholesterol-maleic anhydride via monocarboxylato and O ⁇ Pt bonds, which is significantly better than the complex formed by the combination of cisplatin and macromolecules via stable dicarboxylato bonds, but it is also possible Produces greater toxicity.
  • the researchers prepared platinum anti-tumor drugs into liposomes in order to change the body's distribution in vivo, reduce adverse reactions, and expand the scope of application.
  • cisplatin As an example, its preparation into liposomes faces two major problems.
  • the solubility of cisplatin in water is about 1 mg/mL at 4 °C.
  • cisplatin can only prepare liposomes by passive drug loading, and can not carry out drug loading by pH gradient with reference to the already marketed doxorubicin liposome Doxil. These two reasons usually result in lower drug loading of cisplatin liposomes.
  • SPI-077 developed by ALAZA, is a PEGylated liposome that is primarily heated.
  • the preparation of SPI-077 is increased by a hydration temperature of 65 ° C to increase the solubility of cisplatin by about 8 mg / mL, but the drug loading of cisplatin long-circulating liposomes prepared by this method is still Very low, SPI-077 can only be released after the lipid bilayer is degraded by lipase. Therefore, there are few free drugs released from liposomes and entering the extracellular fluid of the tumor, resulting in a decrease in antitumor activity [Advanced Drug Delivery Reviews, 2013, 65(13-14): 1667-1685].
  • the technical problem to be solved by the present invention is to provide a compound and its use, as well as a platinum complex and a liposome thereof.
  • the compound provided by the invention can be used for the preparation of a platinum complex, and the liposome prepared by using the platinum complex prepared thereby not only significantly improves the antitumor activity, but also significantly reduces its toxicity, especially nephrotoxicity.
  • the prepared liposome platinum complex has high encapsulation efficiency and drug loading amount, and the preparation process is simple and has the value of industrial application.
  • the present invention provides platinum complexes of the structure shown in Formulas I-a to I-b:
  • H attached at any C may be substituted by a substituent
  • substituent is one or more of -NH 2 , -OH, -COOH, halogen or -Ar.
  • R 4 is selected from -CH 2 -, Or removing 2 carboxyl groups of hyaluronic acid
  • the present invention also provides platinum complexes of the structures of Formulas I-c to I-d:
  • H attached at any C may be substituted by a substituent
  • substituent is one or more of -NH 2 , -OH, -COOH, halogen or -Ar.
  • R 6 is selected from -NH 2 , -(CH 2 ) 2 -COOH, -NH-CO-(CH 2 ) e CH 3 or -(CH 2 ) 4 NH-CO-(CH 2 ) 14 CH 3 .
  • the present invention provides an amphiphilic compound of the structure shown in Formula II:
  • H attached at any C may be substituted by a substituent
  • substituent is one or more of -NH 2 , -OH, -COOH halogen or -Ar;
  • R 2 and R 3 includes at least one -COOH.
  • amphiphilic compounds provided by the present invention.
  • R 1 is -C n H 2n+1 and R 2 is R 3 is -OH;
  • R 1 is -C n H 2n+1 , and R 2 is R 3 is -OH;
  • R 1 is -C n H 2n+1
  • R 2 is R 3 is -C a H 2a+1 ;
  • R 1 is -C n H 2n+1
  • R 2 is R 3 is -C a H 2a+1 ;
  • amphiphilic compound provided by the present invention is as shown in Formula II-a to Formula II-b:
  • the platinum complex provided by the present invention is a platinum compound such as cisplatin, carboplatin, oxaliplatin, nedaplatin or lobaplatin (but not limited to the platinum drug) and a carboxyl group in an amphiphilic compound. It is prepared by combining a coordinate bond formed by an amide group.
  • the present invention has no particular requirements for the reaction conditions, and various reaction conditions known in the art which can be used for the preparation of the platinum complex can be used.
  • the amphiphilic compound used in the platinum complex of the formula Ia and the formula Ib of the present invention can be prepared by amidation reaction of a carboxyl group of a dibasic acid (anhydride) with an aliphatic amine (but not limited thereto, all of which can be formed) The synthetic reaction can be).
  • the amphiphilic compound in the platinum complex of the formula Ic and the formula Id is a compound having a similar structure from a fatty acid chloride and an amino acid (such as glutamic acid, aspartic acid, lysine, but not limited thereto).
  • the amidation reaction is prepared.
  • the present invention has no particular requirements on the reaction conditions, and various reaction conditions known in the art can be used for the reaction of an amine with an acid, and an amine with an acid chloride to prepare an amide.
  • the invention also provides a liposome of a platinum complex, comprising the platinum complex, the lipid membrane and/or the stabilizer provided by the invention; the ratio of the platinum complex and the lipid membrane is (99:1) ⁇ (1:99).
  • the ratio is a mass ratio, a molar ratio or a volume ratio.
  • the membrane material comprises a composition of phospholipid and/or cholesterol, wherein the mass fraction of the phospholipid in the membrane is from 1 part to 100 parts, and the mass part of the cholesterol is from 0 part to 60 parts.
  • the phospholipid is egg yolk lecithin, soybean lecithin, hydrogenated soybean phospholipid, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylglycerol, dilauroyl lecithin, and second meat.
  • the present invention provides a medicament for treating cancer, including the liposome provided by the present invention.
  • the platinum complexes and liposomes thereof provided by the present invention may also be prepared into existing dosage forms by adding suitable excipients and additives, such as injection solutions and freeze-dried powder injections, by using techniques well known in the art. , liposome suspensions, tablets, gels, implants, etc., for intravenous, local injection, implantation, inhalation, oral administration.
  • suitable excipients and additives such as injection solutions and freeze-dried powder injections
  • the present invention provides a compound of the formula II, which has good biocompatibility, and has a carboxyl group and an amide group capable of compounding with a platinum drug, and an unstable coordinate bond formed with the platinum drug can be increased.
  • the rate of hydration of the drug in the body increases the efficacy.
  • the platinum complex prepared by the compound provided by the present invention can be well combined with the membrane of the liposome, thereby increasing the encapsulation efficiency and drug loading of the liposome.
  • the carboxyl group has pH sensitivity, and in a lower pH environment (such as tumor tissue), the carboxyl group tends to be deprotonated, which is favorable for promoting the drug in the tumor tissue.
  • the release of the drug improves the efficacy of the drug. Tests have shown that the liposomes provided by the present invention are capable of reducing the toxic side effects of the drug and improving the drug effect.
  • Figure 1-a shows the nuclear magnetic spectrum of the amphiphilic compound of Example 1
  • Figure 1-b shows the nuclear magnetic spectrum of the amphiphilic compound of Example 2
  • Figure 1-c shows the nuclear magnetic spectrum of the amphiphilic compound of Example 3.
  • Figure 1-d-1 shows the infrared spectrum of the amphiphilic compound of Example 4.
  • Figure 1-d-2 shows the nuclear magnetic spectrum of the amphiphilic compound of Example 4.
  • Figure 1-e shows the nuclear magnetic spectrum of the amphiphilic compound of Example 5.
  • Figure 1-f shows the nuclear magnetic spectrum of the amphiphilic compound of Example 9
  • Figure 1-g shows the nuclear magnetic spectrum of the amphiphilic compound of Example 10.
  • Figure 1-h shows the nuclear magnetic spectrum of the amphiphilic compound of Example 11
  • Figure 2-1a shows an infrared spectrum detection spectrum of the platinum complex of Example 12;
  • Figure 2-1b shows an infrared spectrum detection spectrum of the platinum complex of Example 13;
  • Figure 2-3a shows an infrared spectrum detection spectrum of the platinum complex of Example 16
  • Figure 2-3b shows an infrared spectrum detection spectrum of the platinum complex of Example 17
  • Figure 2-4a shows an infrared spectrum detection spectrum of the platinum complex of Example 18
  • Figure 2-4b shows an infrared spectrum detection spectrum of the platinum complex of Example 19;
  • Figure 3 is a diagram showing the electron microscopy of liposomes prepared in Example 31;
  • Figure 4 is a graph showing the particle size distribution of the liposome prepared in Example 31;
  • Figure 5 shows the rate of change in body weight after administration of mice (10 mg/kg).
  • Figure 6 shows the survival rate of mice.
  • the present invention provides a compound and its use, as well as a platinum complex and a liposome thereof, and those skilled in the art can learn from the contents of the present invention and appropriately improve the process parameters. It is to be understood that all such alternatives and modifications are obvious to those skilled in the art and are considered to be included in the present invention.
  • the method and application of the present invention have been described in the preferred embodiments, and it is obvious to those skilled in the art that the present invention can be carried out without departing from the scope, spirit and scope of the invention.
  • the methods and applications of the text are modified or combined and modified to implement and apply the techniques of the present invention.
  • the present invention provides platinum complexes of the structures of Formulas I-a to I-d:
  • H attached at any C may be substituted by a substituent
  • substituent is one or more of -NH 2 , -OH, -COOH, halogen or -Ar.
  • -C n H 2n+1 or -C c H 2c+1 is an alkyl group, and n has a value of from 1 to 22.
  • n has a value of 8-20.
  • n is 8-18 or n is 12-18; in other embodiments, n is 14-16; in some embodiments, n is 15.
  • the value of c and the value of n may be the same or different, and the value of c is 1 to 22.
  • the value of c is 8-20.
  • c is 8-18 or c is 12-18; in other embodiments, c is 14-16; in some embodiments, c is 15.
  • the alkyl group is a linear alkyl group or a branched alkyl group; in some embodiments, it is a linear alkyl group.
  • the alkyl group is a substituted or unsubstituted alkyl group.
  • the substitution means that H attached to any C in the alkyl group may be substituted with a substituent which is one or more of -NH 2 , -OH, -COOH, halogen or -Ar.
  • the number of substituents on any C is one or two.
  • -C n H 2n+1 alkyl includes, but is not limited to, methyl (Me, -CH 3 ), ethyl (Et, -CH 2 CH 3 ), n-propyl (n-Pr, -CH) 2 CH 2 CH 3 ), isopropyl (i-Pr, -CH(CH 3 ) 2 ), n-butyl (n-Bu, -CH 2 CH 2 CH 2 CH 3 ), isobutyl (i-Bu) , -CH 2 CH(CH 3 ) 2 ), sec-butyl (s-Bu, -CH(CH 3 )CH 2 CH 3 ), tert-butyl (t-Bu, -C(CH 3 ) 3 ), positive Pentyl (-CH 2 CH 2 CH 2 CH 2 CH 3 ), 2-pentyl (-CH(CH 3 )CH 2 CH 2 CH 3 ), 3-pentyl (-CH(CH 2 CH 3 ) 2 ) , 2-
  • -C n H 2n-1 or -C c H 2c-1 refers to an alkenyl group in which at least one position is in an unsaturated state, that is, one CC is a sp 2 double bond.
  • the value of n is 2 to 22. In some embodiments, n has a value of 8-20. In other embodiments, n is 8-18 or n is 12-18; in other embodiments, n is 14-16; in some embodiments, n is 15.
  • the value of c and the value of n may be the same or different, and the value of c is 2 to 22. In some embodiments, the value of c is 8-20.
  • c is 8-18 or c is 12-18; in other embodiments, c is 14-16; in some embodiments, c is 15.
  • the alkenyl group is a linear alkenyl group or a branched alkenyl group, and in some embodiments, it is a linear alkenyl group.
  • the alkenyl group is a substituted or unsubstituted alkenyl group. H attached to any C in the substituted alkenyl group may be substituted with a substituent which is one or more of -NH 2 , -OH, -COOH, halogen or -Ar. The number of substituents on any C is one or two.
  • -Ar-C n H 2n+1 means -C n H 2n+1 attached to a phenyl or aryl group
  • -Ar-C n H 2n-1 means attached to a phenyl group or an aryl group.
  • -C n H 2n-1 , -Ar-OC n H 2n+1 means -C n H 2n+1 linked to -Ar-O-, -Ar-OC n H 2n-1 means -Ar- O-phase-linked -C n H 2n-1 .
  • -Ar-C c H 2c + 1 refers to a phenyl or -C c H aryl group connected 2c + 1
  • -Ar-C c H 2c-1 refers to a phenyl or -C c H aryl group connected 2c-1
  • -Ar-OC c H 2c+1 means -C c H 2c+1 linked to -Ar-O-
  • -Ar-OC c H 2c-1 means attached to -Ar-O- -C c H 2c-1 .
  • the -Ar- represents a monocyclic, bicyclic, and tricyclic carbocyclic ring system containing a 6-14 membered ring, and at least one ring system is aromatic, wherein each ring system comprises a 3-7 membered ring, Among them, there are two connection points connected to the rest of the molecule.
  • -Ar- may be a phenylene group, a naphthyl group and an anthranylene group. And the -Ar- may be substituted or unsubstituted.
  • -C b H 2b - or -C d H 2d - means an alkylene group, and b has a value of from 1 to 8. In some embodiments, b has a value of 1 to 6. In other embodiments, b is from 1 to 4; in other embodiments, b is from 2 to 4; the alkylene group is a linear or branched alkylene group; and the value of d and b are taken The values may be the same or different, and the values of d are 1 to 8. In some embodiments, d has a value of 1 to 6.
  • d is from 1 to 4; in other embodiments, d is from 2 to 4; the alkylene group is a linear or branched alkylene group; in some embodiments, Linear alkylene.
  • the alkylene group is a substituted or unsubstituted alkylene group.
  • the substitution means that H attached to any C in the alkylene group may be substituted with a substituent which is one or more of -NH 2 , -OH, -COOH or -Ar.
  • the number of substituents on any C is one or two.
  • alkylene groups include, but are not limited to, methylene (-CH 2 -), ethylene (-CH 2 CH 2 -), n-propylene (-CH 2 CH 2 CH 2 -), Propylene (-C(CH 3 ) 2 -), n-butylene (-CH 2 CH 2 CH 2 CH 2 -), isobutylene, sec-butylene, tert-butylene, n-pentylene, 2-Asia Pentyl, 3-pentylene, 2-methyl-2-butylene, 3-methyl-2-butylene, 3-methyl-1-butylene, 2-methyl-1-butylene, positive Hexylene, 2-hexylene, 3-hexylene, 2-methyl-2-pentylene, 3-methyl-2-pentylene, 4-methyl-2-pentylene, 3-methyl -3-pentylene, 2-methyl-3-pentylene, 2,3-dimethyl-2-butylene, 3,3-dimethyl-2-butylene, n
  • -C b H 2b-2 - or -C d H 2d-2 - means an alkenylene group in which at least one position is in an unsaturated state, that is, one CC is a sp 2 double bond.
  • the value of b is 2 to 8.
  • b has a value of 2-6.
  • b is 2 to 4.
  • the value of d and the value of b may be the same or different, and the value of d is 2 to 8.
  • d has a value of 2-6. In other embodiments, d is from 2 to 4.
  • the alkenyl group is a linear alkenylene group or a branched alkenylene group, and in some embodiments, it is a linear alkenylene group.
  • the alkenylene group is a substituted or unsubstituted alkenylene group. H attached to any C in the substituted alkenylene group may be substituted with a substituent which is one or more of -NH 2 , -OH, -COOH or -Ar. The number of substituents on any C is one or two.
  • -C d H 2d -NH 2 -C d H 2d-2 -NH 2 -C d H 2d is the amino (-NH 2) connected - or -C d H 2d -2 -; -C d H 2d -COOH, -C d H 2d-2 -COOH is -C d H 2d - or -C d H 2d-2 - linked to a carboxyl group (-COOH).
  • R 4 is selected from -CH 2 -, Or removing 2 carboxyl groups of hyaluronic acid
  • R 6 is selected from -NH 2 , -(CH 2 ) 4 NH 2 , -(CH) 2 -COOH or -(CH 2 ) 4 NH-CO-(CH 2 ) 14 CH 3 .
  • the platinum complex provided by the invention is prepared by reacting a compound of formula II or a compound of formula III with a platinum compound; the platinum compound is cisplatin, carboplatin, oxaliplatin or nedaplatin, iriplatin, lobaplatin Wait.
  • H attached at any C may be substituted by a substituent
  • substituent is one or more of -NH 2 , -OH, -COOH, halogen or -Ar;
  • R 2 and R 3 includes at least one -COOH.
  • the molar ratio of the compound of formula II or formula III to the platinum compound is not more than 30:1.
  • the molar ratio of the compound of Formula II or Formula III to Pt in the platinum compound is (1-8):1.
  • the molar ratio of the compound of Formula II or Formula III to the Pt in the platinum compound is (1 to 5):1.
  • the molar ratio of the compound of Formula II or Formula III to the Pt in the platinum compound is 1:1.
  • the solvent used in the reaction for preparing the platinum complex is an aqueous medium.
  • the aqueous medium is water or mannitol aqueous solution or sodium chloride aqueous solution or glycerin aqueous solution or sugar-containing aqueous solution or phosphate buffer solution (not limited thereto), and may also contain dimethyl group.
  • the present invention provides an amphiphilic compound of the structure shown in Formula II:
  • H attached at any C may be substituted by a substituent
  • substituent is one or more of -NH 2 , -OH, -COOH, halogen or -Ar;
  • R 2 and R 3 includes at least one -COOH.
  • -C n H 2n+1 or -C a H 2a+1 is an alkyl group, and n has a value of from 1 to 22.
  • n has a value of 8-20.
  • n is 8-18 or n is 12-18; in other embodiments, n is 14-16; in some embodiments, n is 15.
  • the value of a and the value of n may be the same or different, and the value of a is 1 to 22.
  • a has a value of 8-20.
  • a is 8-18 or a is 12-18; in other embodiments, a is 14-16; in some embodiments, a is 15.
  • the alkyl group is a linear alkyl group or a branched alkyl group; in some embodiments, it is a linear alkyl group.
  • the alkyl group is a substituted or unsubstituted alkyl group.
  • the substitution means that H attached to any C in the alkyl group may be substituted with a substituent which is one or more of -NH 2 , -OH, -COOH or -Ar.
  • the number of substituents on any C is one or two.
  • -C n H 2n+1 alkyl includes, but is not limited to, methyl (Me, -CH 3 ), ethyl (Et, -CH 2 CH 3 ), n-propyl (n-Pr, -CH) 2 CH 2 CH 3 ), isopropyl (i-Pr, -CH(CH 3 ) 2 ), n-butyl (n-Bu, -CH 2 CH 2 CH 2 CH 3 ), isobutyl (i-Bu) , -CH 2 CH(CH 3 ) 2 ), sec-butyl (s-Bu, -CH(CH 3 )CH 2 CH 3 ), tert-butyl (t-Bu, -C(CH 3 ) 3 ), positive Pentyl (-CH 2 CH 2 CH 2 CH 2 CH 3 ), 2-pentyl (-CH(CH 3 )CH 2 CH 2 CH 3 ), 3-pentyl (-CH(CH 2 CH 3 ) 2 ) , 2-
  • -C n H 2n-1 or -C a H 2a-1 refers to an alkenyl group in which at least one position is in an unsaturated state, that is, one CC is a sp 2 double bond.
  • the value of n is 2 to 22. In some embodiments, n has a value of 8-20. In other embodiments, n is 8-18 or n is 12-18; in other embodiments, n is 14-16; in some embodiments, n is 15.
  • the value of a and the value of n may be the same or different, and the value of a is 2 to 22. In some embodiments, a has a value of 8-20.
  • a is 8-18 or a is 12-18; in other embodiments, a is 14-16; in some embodiments, a is 15.
  • the alkenyl group is a linear alkenyl group or a branched alkenyl group, and in some embodiments, it is a linear alkenyl group.
  • the alkenyl group is a substituted or unsubstituted alkenyl group. H attached to any C in the substituted alkenyl group may be substituted with a substituent which is one or more of -NH 2 , -OH, -COOH, halogen or -Ar. The number of substituents on any C is one or two.
  • -Ar-C n H 2n+1 means -C n H 2n+1 attached to a phenyl or aryl group
  • -Ar-C n H 2n-1 means attached to a phenyl group or an aryl group.
  • -C n H 2n-1 , -Ar-OC n H 2n+1 means -C n H 2n+1 linked to -Ar-O-, -Ar-OC n H 2n-1 means -Ar- O-phase-linked -C n H 2n-1 .
  • -Ar-C a H 2a+1 means -C a H 2a+1 attached to a phenyl or aryl group
  • -Ar-C a H 2a-1 means -C a H attached to a phenyl or aryl group 2a-1
  • -Ar-OC a H 2a+1 means -C a H 2a+1 linked to -Ar-O-
  • -Ar-OC a H 2a-1 means attached to -Ar-O- -C a H 2a-1 .
  • the -Ar- represents a monocyclic, bicyclic, and tricyclic carbocyclic ring system containing a 6-14 membered ring, and at least one ring system is aromatic, wherein each ring system comprises a 3-7 membered ring, Among them, there are two connection points connected to the rest of the molecule.
  • -Ar- may be a phenylene group, a naphthyl group and an anthranylene group. And the -Ar- may be substituted or unsubstituted.
  • -C m H 2m - means an alkylene group, and m has a value of from 1 to 8. In some embodiments, m has a value of 1 to 6. In other embodiments, m is from 1 to 4; in other embodiments, m is from 2 to 4; the alkylene group is a linear or branched alkylene group; in some embodiments, Linear alkylene.
  • the alkylene group is a substituted or unsubstituted alkylene group.
  • the substitution means that H attached to any C in the alkylene group may be substituted with a substituent which is one or more of -NH 2 , -OH, -COOH or -Ar.
  • the number of substituents on any C is one or two.
  • -C m H 2m -alkylene includes, but is not limited to, methylene (-CH 2 -), ethylene (-CH 2 CH 2 -), n-propylene (-CH 2 CH 2 CH 2 -), isopropylidene (-C(CH 3 ) 2 -), n-butylene (-CH 2 CH 2 CH 2 CH 2 -), isobutylene, sec-butylene, tert-butylene, Ortho Pentyl, 2-pentylene, 3-pentylene, 2-methyl-2-butylene, 3-methyl-2-butylene, 3-methyl-1-butylene, 2-methyl- 1-butylene, n-hexylene, 2-hexylene, 3-hexylene, 2-methyl-2-pentylene, 3-methyl-2-pentylene, 4-methyl-2-pentylene , 3-methyl-3-pentylene, 2-methyl-3-pentylene, 2,3-dimethyl-2-butylene, 3,3-di
  • -C m H 2m-2 - refers to an alkenylene group in which at least one position is in an unsaturated state, that is, one CC is a sp 2 double bond.
  • the value of n is 2 to 8. In some embodiments, n has a value of 2-6. In other embodiments, n is from 2 to 4.
  • the alkenyl group is a linear alkenylene group or a branched alkenylene group, and in some embodiments, it is a linear alkenylene group.
  • the alkenylene group is a substituted or unsubstituted alkenylene group.
  • H attached to any C in the substituted alkenylene group may be substituted with a substituent which is one or more of -NH 2 , -OH, -COOH or -Ar.
  • the number of substituents on any C is one or two.
  • -C m H 2m -NH- or -C m H 2m-2 -NH- means -C m H 2m - or -C m H 2m-2 - linked to -NH-.
  • H attached to any C may be substituted by one or two substituents
  • substituent is -NH 2 or -COOH.
  • R 2 and R 3 includes at least one -COOH.
  • R 1 is -C n H 2n+1 and R 2 is R 3 is -OH;
  • R 1 is -C n H 2n+1 ;
  • R 2 is R 3 is -OH;
  • R 1 is -C n H 2n+1
  • R 2 is R 3 is -C a H 2a+1 ;
  • R 1 is -C n H 2n+1
  • R 2 is R 3 is -C a H 2a+1 ;
  • the structure is:
  • the platinum-based complex provided by the present invention can bind well to the membrane of the liposome, thereby increasing the encapsulation efficiency and the drug loading amount of the liposome. Moreover, after preparing the platinum complex with the compound provided by the invention, the carboxyl group has pH sensitivity, and in a lower pH environment (such as tumor tissue), the carboxyl group tends to be deprotonated, which is favorable for promoting the drug in the tumor tissue. The release of the drug improves the efficacy of the drug and reduces the side effects of the drug.
  • the invention also provides a liposome of a platinum complex, comprising the platinum complex, the lipid membrane and/or the stabilizer provided by the invention; the ratio of the platinum complex and the lipid membrane is (99:1) ⁇ (1:99).
  • the ratio is a mass ratio, a molar ratio or a volume ratio.
  • the lipid membrane material comprises a composition of phospholipid and/or cholesterol, wherein the mass fraction of the phospholipid in the membrane is from 1 part to 100 parts, and the mass part of the cholesterol is from 0 part to 60 parts;
  • the phospholipid is egg yolk lecithin, soybean lecithin, hydrogenated soybean phospholipid, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylglycerol, dilauroyl lecithin, dimyristoyl egg Phospholipid, dipalmitoyl lecithin or distearoyl lecithin, dioleoylphosphatidylcholine, dilauroylphosphatidylglycerol, dimyristoylphosphatidylglycerol, dipalmitoylphosphatidylglycerol or distearoylphospholipid Any one or a combination of two or more of acylglycerol, dioleoylphosphatidylglycerol, phosphoacylglycerol, di-erucylphosphatidylglycerol, and PEGylated phospholipid.
  • the liposome provided by the present invention or also includes a long circulating material.
  • the long circulating materials used in the present invention are MPEG2000-DSPE, MPEG5000-DSPE, MPEG2000-DMPE, MPEG5000-DMPE, MPEG2000-DPPE, MPEG5000-DPPE and the like.
  • the mass fraction of the long circulating material in the liposome provided by the invention is 0-50%.
  • the liposome provided by the present invention may further comprise an antioxidant; the antioxidant used in the present invention is L-cysteine, ascorbic acid, dl-strontium-tocopherol, sodium sulfite, sodium metabisulfite or anhydrous sodium hydrogen sulfite.
  • an antioxidant used in the present invention is L-cysteine, ascorbic acid, dl-strontium-tocopherol, sodium sulfite, sodium metabisulfite or anhydrous sodium hydrogen sulfite.
  • the antioxidant has a mass fraction of not more than 20%.
  • An isotonicity adjusting agent may be further added to the liposome provided by the present invention, and the isotonicity adjusting agent is a composition of any one or more of mannitol, sorbitol, glucose, sucrose or sodium chloride.
  • the invention provides a method for preparing a liposome, which can adopt a film dispersion method, a reverse evaporation method or Double emulsion method, etc.
  • the liposome is prepared by placing a membrane, an antioxidant, a long-circulating material in a round bottom flask, adding an appropriate amount of an organic solvent to dissolve, and adding an aqueous solution of the platinum complex provided by the present invention. , mechanical or ultrasonic homogenization to form W / O emulsion, rotary evaporation to remove organic solvent, forming a film on the wall of the container, injecting hydration medium, so that the membrane is hydrated for 1h ⁇ 2h, through the liposome extruder or evenly A novel platinum-like liposome having a uniform particle size is obtained, and the long-circulating material can also be added after the liposome is formed, stirred, and modified on the surface of the liposome.
  • the liposome is prepared by taking a film, an antioxidant, a platinum-based compound and a long-circulating material provided by the present invention, and placing it in a round bottom flask, adding an appropriate amount of an organic solvent to dissolve, and rotating under reduced pressure.
  • the organic solvent was removed by evaporation, a film was formed on the wall of the container, and a hydration medium was injected to hydrate the film for 1 h to 2 h, and the new platinum-like liposome was obtained by a liposome extruder or homogenization.
  • the liposome is prepared by placing a membrane, an antioxidant, and a long-circulating material in a round bottom flask, adding an appropriate amount of an organic solvent to dissolve, adding the platinum complex provided by the present invention, and performing ultrasonication. Or mechanically homogenizing to form a W/O emulsion, removing some organic solvent by rotary evaporation under reduced pressure, injecting into a hydration medium, mechanically stirring to form W/O/W, and then removing the organic solvent by rotary evaporation under reduced pressure, passing through a liposome extruder or Homogenization gave novel platinum-based liposomes.
  • the present invention also provides a medicament for treating cancer, comprising the liposome and preparation provided by the present invention.
  • the medicament provided by the present invention can be used for the treatment of cancer, such as breast cancer, non-small cell lung cancer, testicular cancer, pancreatic cancer, rectal cancer, liver cancer, ovarian cancer or head and neck cancer.
  • cancer such as breast cancer, non-small cell lung cancer, testicular cancer, pancreatic cancer, rectal cancer, liver cancer, ovarian cancer or head and neck cancer.
  • the pharmaceutical preparations provided by the present invention are injections (suspension or lyophilized powder for injection), emulsions, submicroemulsions and the like.
  • the test shows that the liposome provided by the invention can significantly increase the blood concentration of the platinum compound, prolong the residence time of the drug in the body, and can concentrate the platinum drug in the cancerous organ, thereby not only reducing the dosage and increasing the therapeutic effect. Moreover, it can reduce the toxic side effects of drugs and is suitable for the treatment of various tumor diseases.
  • the present invention provides a compound of the formula II, which has good biocompatibility, and has a carboxyl group and an amide group capable of compounding with a platinum drug, and an unstable coordinate bond formed with the platinum drug can be increased.
  • the rate of hydration of the drug in the body increases the efficacy.
  • the platinum complex prepared by the compound provided by the invention can be well combined with the membrane of the liposome, thereby increasing the encapsulation efficiency and drug loading of the liposome.
  • the carboxyl group has pH sensitivity, and in a lower pH environment (such as tumor tissue), the carboxyl group tends to be deprotonated, which is favorable for promoting the drug in the tumor tissue.
  • the release further enhances the efficacy of the drug and reduces its side effects.
  • the liposome provided by the present invention can significantly reduce the toxic side effects of the drug and improve the drug effect.
  • the reagents or instruments used in the present invention are all commercially available, and are commercially available.
  • the preparation method is as follows:
  • Example 1 After adding 1.5 mmol of fatty amine (such as octadecylamine, hexadecylamine, tetradecylamine or dodecylamine) to 20 mL of chloroform, another 3.3 mmol of an acid anhydride (such as maleic anhydride or succinic anhydride) is dissolved in 20 mL of chloroform. After dissolving, the chloroform solution of the fatty amine is added to the acid anhydride solution, stirred for 30 minutes, filtered under reduced pressure, and the obtained product is washed successively with chloroform, acetone, and dried (molar ratio of 1:2) to obtain the above ( Example 1 - Example 5) Amphiphilic compound.
  • the nuclear magnetic structure was characterized for the amphiphilic compound, and the results are shown in Figures 1-a to 1-e.
  • the preparation method is:
  • the preparation method is as follows:
  • the water-washed material was oven-dried at 50 ° C, taken out, washed three times with a mixed solvent of petroleum ether: acetone: ethyl acetate (100:1:1), centrifuged, taken out, and dried under reduced pressure.
  • the prepared compound was subjected to nuclear magnetic structure characterization, and the results are shown in Fig. 1-g.
  • the preparation method is as follows:
  • the preparation method comprises the following steps: adding an amphiphilic compound to water and stirring and dissolving under certain conditions, adding a platinum compound (molar ratio of the amphiphilic compound to the platinum compound: 1:1-10:1), heating and stirring for 12 hours, the reaction is obtained.
  • the platinum complex is dried to obtain a solid powder of the platinum complex.
  • the infrared detection spectrum is shown in Figure 2-1a to Figure 2-9b.
  • the amphiphilic compound forms a complex with oxaliplatin.
  • organic solvent chloroform or water-saturated ether or dichloromethane, methanol, ethanol or a mixed solvent of at least two solvents
  • mechanical or ultrasonic homogenization to form W / O emulsion
  • rotary evaporation to remove organic solvent
  • Form a film on the wall of the container inject a hydration medium (physiological saline or 5% dextrose or water for injection or mannitol aqueous solution), and hydrate the film at 40 ° C (or 55 ° C) for 1 h to 2 h, through the liposome extruder Or micro-fluid homogenization to obtain platinum complex liposomes, adding long-circulation material and stirring at room temperature for 1 h, which
  • the formulated amount of the membrane and the antioxidant, the long circulating material, and the platinum compound added in the formula amount are placed in a round bottom flask, dissolved in an appropriate amount of chloroform, and the organic solvent is removed by rotary evaporation under reduced pressure to form a film on the wall of the container.
  • the hydration medium water for injection
  • the membrane was hydrated at 40-55 ° C for 1 to 2 h, and homogenized by a liposome extruder to obtain a novel platinum-based liposome.
  • the morphology of the platinum-containing complex liposome prepared by the present invention was observed by an electron microscope, and the particle size and distribution of the liposome were measured by a Zetasizer Nano ZS90 laser particle size analyzer.
  • the encapsulation efficiency of the liposome produced by the present invention was calculated by the method provided by the Chinese Pharmacopoeia. The results are shown in Table 5:
  • the novel platinum-based complex liposome preparation provided by the invention has a particle size of 90-150 nm, an encapsulation efficiency of 92-98%, and a drug loading of 8-30%. Unlike the liposomes reported in the prior literature and patents, the platinum liposome has the characteristics of high encapsulation efficiency, good stability and strong anticancer activity.
  • the liposomes prepared by using commercially available cisplatin for injection and the samples of Examples 31, 33 and 36 were used as experimental materials, and the MCF-7, A549 and HEPG2 cells were digested with trypsin, and then centrifuged and collected. The cell suspension was adjusted and the cell count was adjusted to a concentration of 5-10 x 10 4 /mL. After the cell suspension was prepared, gently mix, add 100 ⁇ L per well, and place the inoculated cells in an incubator for 24 h, respectively, adding 1 ⁇ mol/L, 2 ⁇ mol/L, 4 ⁇ mol/L, 8 ⁇ mol/L, 16 ⁇ mol.
  • liver cancer cells H22 with logarithmic growth were adjusted to a single cell concentration of (1-5) x 10 6 cells/mL.
  • the H22 cell suspension was inoculated subcutaneously into the right axilla of the mouse, each 0.2 mL, and a subcutaneous inoculation model of the liver cancer H22 tumor strain was established.
  • Tumor-bearing mice were randomly divided into 5 groups, 10 in each group, respectively, saline group, cisplatin control group for injection, and experimental group (platinum complex lipids provided in Examples 31, 33 and 36 of the present invention, respectively).
  • the plastids were labeled in the group.
  • the rats in each group were injected with normal saline, cisplatin control group and experimental group. They were administered once every three days and twice, and the dose was 2.5 mg/kg.
  • the survival state of the mice was observed daily, and the mice were sacrificed on the 7th day after the administration, the tumor was taken out, the surface was washed with physiological saline, the filter paper was blotted, and the tumor inhibition rate was calculated, and the calculation method was as follows:
  • Tumor inhibition rate (%) (average tumor weight of the control group - mean tumor weight of the test group) ⁇ average tumor weight of the control group ⁇ 100
  • mice were randomly divided into groups, 5 rats in each group, which were blank control group, cisplatin control group for injection, liposomes prepared in Examples 31, 33 and 36, and the groups were labeled with tail vein injection.
  • the liposomes prepared in the physiological saline, the commercially available cisplatin control group, and the examples 31, 33, and 36 were administered once, and the doses were 10 mg/kg and 20 mg/kg, respectively, after administration, daily.
  • the survival state of the mice was observed, the body weight was weighed, and the survival rate of the mice was recorded.
  • mice were randomly divided into 5 groups, 10 in each group, blank control group, respectively, examples Liposomes prepared in 31, 33, and 36, commercially available cisplatin control group for injection, labeling in the group, intravenous injection of physiological saline in each group, and liposome solutions prepared in Examples 31, 33, and 36, cisplatin
  • the injection was administered once, and the dose was 10 mg/kg.
  • the survival state of the mice was observed daily, and the mice were sacrificed on the 7th day after administration, and the urea nitrogen and creatinine values of the mice were measured, and the results are shown in Table 8:
  • the experimental results show that the cisplatin complex liposome prepared by the present invention can significantly reduce the nephrotoxicity of cisplatin compared with the cisplatin injection.

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Abstract

一种铂类配合物、其脂质体及其应用。该铂类配合物含有羧基具有pH敏感性，在较低的pH值环境中(如肿瘤组织)，羧基趋于去质子化，有利于促进药物在肿瘤组织的释放，提高药物的疗效，且能够与脂质体的膜材良好的结合，从而提高脂质体的包封率和载药量。实验表明，该铂类配合物的脂质体能够降低药物的毒副作用并提高药效。

Description

一种化合物及其应用以及一种铂类配合物及其脂质体
本申请要求于2016年01月25日提交中国专利局、申请号为201610049404.9、发明名称为“一种化合物及其应用以及一种铂类配合物及其脂质体”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
本发明涉及医药领域，尤其涉及一种化合物及其应用以及一种铂类配合物及其脂质体。
背景技术
自1965年美国科学家B.Rosenborg偶然发现顺铂具有抗肿瘤活性以来，铂类金属抗癌药物的合成应用和研究得到了迅速的发展，与传统的细胞毒类抗肿瘤药物相比，铂类抗肿瘤药物的作用机制独特。其主要作用靶点为DNA，当铂类抗肿瘤药物进入细胞内部后被水解，形成具有高活性的带正电的水合铂，水合铂与核酸和蛋白质反应，形成DNA链内链间共轭物，从而导致DNA的二级结构发生变化，阻碍DNA的转录与复制，最终导致细胞凋亡。目前相继成功开发了顺式-二氯二胺铂即顺铂(Cisplatin)、顺式-1,1-环丁烷二羧酸二胺铂即卡铂(Carboplatin)、顺式-草酸-(反式-(-)-1,2-环己二胺)合铂即奥沙利铂(Oxaliplatin)、奈达铂(Nedaplatin)、依铂(Eptaplatin)、洛铂(Lobaplatin)等。铂类抗肿瘤药物具有抗癌谱广、活性强、无交叉耐药性等特点。有统计数据表明，在所有的化疗方案中有70％～80％以铂为主或有铂类药物参与配伍。然而，铂类抗癌药物对肿瘤细胞缺乏特异性，静脉注射后，在血液中迅速被消除，快速分布到各个脏器组织，尤其是在肝、肾、肠等组织分布较多，因此产生严重的毒副作用。主要包括肾毒性，消化道毒性(恶心、呕吐)，骨髓毒性，神经毒性和耳毒性等，且存在到达肿瘤部位的药物浓度较低、降低其疗效，长期应用还会使癌细胞产生耐药性、从而降低其化疗作用等缺点。
为了解决上述问题，延长药物在血液中的循环时间，减少药物与蛋 白间的非特异性结合，从而降低其全身毒副作用，提高药效。各国药学研究者已致力于铂类药物靶向给药系统的研究，如：胶束、囊泡、脂质体、纳米粒等。这些药物载体能够有效的将药物包载于其中，并利用载体的性质，实现药物的输送和控制释放，从而提高药物在靶部位的有效聚集，降低非病灶部位的药物浓度，不仅可以提高疗效，还可以降低其全身的毒副作用。
现有技术公开了多种载体担载药物的方式开发铂类制剂的方法。如Uchino等人利用将顺铂与富含羧基的聚乙二醇-b-聚谷氨酸复合制备了载药量为30wt.％的胶束(NC-6004)已经进入II期临床研究【British Journal of Cancer(2005)93,678-687】,然而由于其所得的胶束是聚氨基酸侧链间的交联作用而形成，此方法所得的复合物经冻干后的冻干粉会很难复溶，且该制剂疗效无提高。Access Pharmaceuticals公司将常用的聚合物HPMA与顺铂相结合制备了AP5280，AP5280的肿瘤聚集量是顺铂注射剂的19倍且显著的降低了顺铂的肾毒性【J.Drug Target 10(2002)549-556】。这些研究表明铂类化合物可以通过环境敏感的化学键与大分子结合，从而有效的降低铂类药物的毒性，提高疗效。而且不同的环境敏感的化学键对铂类抗癌药物的药效至关重要，因其能够改变铂类化合物在体内的水合速度，铂类化合物在体内的水合速度越快，抗肿瘤效果越强，但与此同时其毒性也越大。Poulomi Sengupta等人将顺铂与胆固醇-马来酸酐通过monocarboxylato和O→Pt键结合产生的药效要明显优于顺铂与大分子通过稳定的dicarboxylato键结合形成的复合物，但由此也可能产生较大的毒性。为此，研究者将铂类抗肿瘤药物制备成脂质体以期改变药物的体内分布，减轻不良反应，拓展应用范围。
然而铂类抗肿瘤药物制备脂质体面临很大的困难。以顺铂为例，将其制备成脂质体面临两大问题，首先顺铂在水中的溶解度4℃时约为1mg/mL。其次，顺铂只能通过被动载药的方式制备脂质体，无法参照已经上市的阿霉素脂质体Doxil等进行pH梯度主动载药。上述两个原因通常造成顺铂脂质体的载药量较低。由ALAZA公司研究开发的顺铂长循环脂质体(SPI-077)，是一种PEG化修饰的脂质体，其主要通过加热的方 式增加顺铂的溶解度，SPI-077的制备过程采用水化温度为65℃的高温使顺铂的溶解度提高约8mg/mL，但是该方法制备的顺铂长循环脂质体的载药量依然很低，SPI-077只能通过脂肪酶降解脂质双分子后，药物才能释放出来。因此从脂质体中释放出来并进入肿瘤细胞外液的游离药物很少，导致抗肿瘤活性降低【Advanced Drug Delivery Reviews,2013,65(13-14):1667–1685】。
可见，现有的技术仍不能很好的解决铂类化合物存在的上述问题，无法在提高载药量的同时维持药物良好的活性且又能降低其毒副作用。
发明内容
有鉴于此，本发明要解决的技术问题在于提供一种化合物及其应用以及一种铂类配合物及其脂质体。本发明提供的化合物能够用于铂类配合物的制备，用其制备的铂类配合物制备而得的脂质体既显著提高了抗肿瘤活性，又显著降低了其毒性，特别是肾毒性，而且所制备的脂质体铂类配合物的包封率和载药量均较高，其制备工艺简单，具有产业化应用的价值。
本发明提供了式I-a～I-b所示结构的铂类配合物：
Figure PCTCN2017071114-appb-000001
R4选自-CbH2b-、-CbH2b-2-、-Ar-CbH2b-、-Ar-CbH2b-2-、-Ar-O-CbH2b-、-Ar-O-CbH2b-2-或除去2个羧基的透明质酸,b＝0～22；
R5选自-H、-CcH2c+1、-CcH2c-1、-Ar-CcH2c+1、-Ar-CcH2c-1、-Ar-O-CcH2c+1、-Ar-O-CcH2c-1或除去1个氨基的壳聚糖,c＝1～22；
其中，任意C上连接的H可以被取代基所取代；
其中，所述取代基为-NH2、-OH、-COOH、卤素或-Ar中的一种或几种。
在本发明的实施例中，
R4选自-CbH2b-、-CbH2b-2-或除去2个羧基的透明质酸,b＝1～10；
R5选自-CcH2c+1、-CcH2c-1或除去1个氨基的壳聚糖,c＝8～18；
在本发明的实施例中，
R4选自-CH2-、
Figure PCTCN2017071114-appb-000002
Figure PCTCN2017071114-appb-000003
或除去2个羧基的透明质酸；
R5选自-CcH2c+1，c＝12～18。
本发明还提供了式I-c～I-d所示结构的铂类配合物：
Figure PCTCN2017071114-appb-000004
R1选自-H、-CnH2n+1、-CnH2n-1、-Ar-CnH2n+1、-Ar-CnH2n-1、-Ar-O-CnH2n+1或-Ar-O-CnH2n-1,n＝1～22；
R6选自-NH2、-NH-CO-(CH2)eCH3、-(CH2)dNH-CO-(CH2)eCH3，-CdH2d-NH2、-CdH2d-2-NH2、-CdH2d-COOH、-CdH2d-2-COOH，d＝1～8,e＝1～21；
x＝0～10；
其中，任意C上连接的H可以被取代基所取代；
其中，所述取代基为-NH2、-OH、-COOH、卤素或-Ar中的一种或几种。
在本发明的实施例中，
x＝0～10；
R1选自-H、-CnH2n+1或-CnH2n-1,n＝8～20；
R6选自-NH2、-NH-CO-(CH2)eCH3、-CdH2d-NH2、-CdH2d-2-COOH或-(CH2)dNH-CO-(CH2)eCH3；d＝1～6,e＝8～20。
在本发明的实施例中，
x＝0～6；
R1为-CnH2n+1，n＝8～18；
R6选自-NH2、-(CH2)2-COOH、-NH-CO-(CH2)eCH3或-(CH2)4NH-CO-(CH2)14CH3。
在一些实施例中，x＝0；R1为-CnH2n+1，n＝8～18；R6为-(CH2)2-COOH、-(CH2)4NH2或-(CH2)4NH-CO-(CH2)14CH3。
x＝1～6；R1为-CnH2n+1，n＝8～18；R6为-NH2。
本发明提供了式II所示结构的两亲性化合物：
Figure PCTCN2017071114-appb-000005
R1选自-H、-CnH2n+1、-CnH2m-1、-Ar-CnH2n+1、-Ar-CnH2n-1、-Ar-O-CnH2n+1或-Ar-O-CnH2n-1,n＝1～22；
R2选自-CmH2m-、-CmH2m-2-、-CmH2m-NH-或-CmH2m-2-NH-,m＝1～8；
R3选自-OH、-CaH2a+1、-CaH2a-1、-Ar-CaH2a+1、-Ar-CaH2a-1、-Ar-O-CaH2a+1或-Ar-O-CaH2a-1,a＝1～22；
其中，任意C上连接的H可以被取代基所取代；
其中，所述取代基为-NH2、-OH、-COOH卤素或-Ar中的一种或几种；
所述R2和R3的结构中至少包括1个-COOH。
本发明提供的两亲性化合物中，
R1为-CnH2n+1，R2为
Figure PCTCN2017071114-appb-000006
R3为-OH；
或R1为-CnH2n+1，R2为
Figure PCTCN2017071114-appb-000007
R3为-OH；
或R1为-CnH2n+1，R2为
Figure PCTCN2017071114-appb-000008
R3为-CaH2a+1；
或R1为-CnH2n+1，R2为
Figure PCTCN2017071114-appb-000009
R3为-CaH2a+1；
n＝8～18；m＝2～6；a＝8～18。
本发明提供的两亲性化合物的结构如式II-a～式II-b所示：
Figure PCTCN2017071114-appb-000010
本发明提供的铂类配合物是由顺铂、卡铂、奥沙利铂、奈达铂或洛铂(但不仅限于此铂类药物)等铂类化合物与一种两亲性化合物中的羧基与酰胺基团形成的配位键结合而制备。本发明对反应条件没有特殊的要求，本领域公知的可用于制备铂类配合物的反应条件均可。
本发明式I-a、式I-b铂类配合物中使用的两亲性化合物可以由二元酸(酸酐)中的一个羧基与脂肪胺发生酰胺化反应而制备(但不仅限于此，所有可生成该结构的合成反应均可)。本发明式I-c、式I-d铂类配合物中的两亲性化合物是由脂肪酰氯与氨基酸(如谷氨酸、天冬氨酸、赖氨酸，但不仅限于此，有类似结构的化合物均可)发生酰胺化反应制备而得。本发明对反应条件没有特殊的要求，本领域公知的可用于胺与酸，胺与酰氯反应制备酰胺的反应条件均可。
本发明还提供了一种铂类配合物的脂质体，包括本发明提供的铂类配合物、脂质膜材和/或稳定剂；所述铂类配合物、脂质膜材的比例为(99：1)～(1：99)。
所述比例为质量比、摩尔比或体积比。
本发明提供的脂质体中，膜材包括磷脂和/或胆固醇的组合物，膜材中磷脂的质量份为1份～100份，胆固醇的质量份为0份～60份。
本发明提供的脂质体中，所述磷脂为蛋黄卵磷脂、大豆卵磷脂、氢化大豆磷脂、磷脂酰丝氨酸、磷脂酰肌醇、磷脂酰乙醇胺、磷脂酰甘油、二月桂酰卵磷脂、二肉豆蔻酰基卵磷脂、二棕榈酰基卵磷脂或二硬脂酰基卵磷脂、二油酰基磷脂酰胆碱、二月桂酰磷脂酰甘油、二肉豆蔻酰基磷脂酰甘油、二棕榈酰基磷脂酰甘油或二硬脂酰基磷脂酰甘油、二油酰基磷脂酰甘油、磷酸酰甘油、二芥酰磷脂酰甘油、PEG化磷脂等中的任一种或两者以上的组合物。
本发明提供了一种治疗癌症的药物，包括本发明提供的脂质体。
本发明提供的铂类配合物及其脂质体，还可以加入适宜的辅料和附加剂利用本领域公知的技术制备成现有不同剂型(dosage form)，如注射液、注射用冻干粉针，脂质体混悬液、片剂、凝胶剂、植入剂等，供静脉注射、局部注射、植入、吸入、口服等途径给药。
本发明提供了式II结构的化合物，该化合物具有良好的生物相容性，其具有的羧基与酰胺基团能够与铂类药物配合，其与铂类药物形成的不稳定的配位键能够增加药物在体内的水合速度从而提高药效。同时，以本发明提供的化合物制得的铂类配合物能够与脂质体的膜材良好的结合，从而提高脂质体的包封率和载药量。并且，以本发明提供的化合物制备铂类配合物后，含有羧基具有pH敏感性，在较低的pH值环境中(如肿瘤组织)，羧基趋于去质子化，有利于促进药物在肿瘤组织的释放，提高药物的疗效。试验表明，本发明提供的脂质体能够降低药物的毒副作用并提高药效。
附图说明
图1-a示实施例1两亲性化合物的核磁图谱；
图1-b示实施例2两亲性化合物的核磁图谱；
图1-c示实施例3两亲性化合物的核磁图谱；
图1-d-1示实施例4两亲性化合物的红外图谱；
图1-d-2示实施例4两亲性化合物的核磁图谱；
图1-e示实施例5两亲性化合物的核磁图谱；
图1-f示实施例9两亲性化合物的核磁图谱；
图1-g示实施例10两亲性化合物的核磁图谱；
图1-h示实施例11两亲性化合物的核磁图谱；
图2-1a示实施例12铂类配合物的红外光谱检测图谱；
图2-1b示实施例13铂类配合物的红外光谱检测图谱；
图2-2a示实施例14铂类配合物的红外光谱检测图谱；
图2-2b示实施例15铂类配合物的红外光谱检测图谱；
图2-3a示实施例16铂类配合物的红外光谱检测图谱；
图2-3b示实施例17铂类配合物的红外光谱检测图谱；
图2-4a示实施例18铂类配合物的红外光谱检测图谱；
图2-4b示实施例19铂类配合物的红外光谱检测图谱；
图2-5a示实施例20铂类配合物的红外光谱检测图谱；
图2-5b示实施例21铂类配合物的红外光谱检测图谱；
图2-6a示实施例22铂类配合物的红外光谱检测图谱；
图2-7a示实施例23铂类配合物的红外光谱检测图谱；
图2-7b示实施例24铂类配合物的红外光谱检测图谱；
图2-8a示实施例25铂类配合物的红外光谱检测图谱；
图2-9a示实施例26铂类配合物的红外光谱检测图谱；
图2-9b示实施例27铂类配合物的红外光谱检测图谱；
图3示实施例31制得脂质体的电镜检测图；
图4示实施例31制得脂质体的粒径分布图；
图5示小鼠给药后体重变化率(10mg/kg)；
图6示小鼠存活率。
具体实施方式
本发明提供了一种化合物及其应用以及一种铂类配合物及其脂质体，本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本 文的方法和应用进行改动或适当变更与组合，来实现和应用本发明技术。本发明提供了式I-a～I-d所示结构的铂类配合物：
Figure PCTCN2017071114-appb-000011
R1选自-H、-CnH2n+1、-CnH2n-1、-Ar-CnH2n+1、-Ar-CnH2n-1、-Ar-O-CnH2n+1或-Ar-O-CnH2n-1；n＝1-22；
R4选自-CbH2b-、-CbH2b-2-、-Ar-CbH2b-、-Ar-CbH2b-2-、-Ar-O-CbH2b-、-Ar-O-CbH2b-2-或除去2个羧基的透明质酸,b＝0～22；
R5选自-H、-CcH2c+1、-CcH2c-1、-Ar-CcH2c+1、-Ar-CcH2c-1、-Ar-O-CcH2c+1、-Ar-O-CcH2c-1或除去1个氨基的壳聚糖,c＝0～21；
R6选自-NH2、-CdH2d-NH2、-CdH2d-2-NH2、-CdH2d-COOH、-CdH2d-2-COOH、-NH-CO-(CH2)eCH3或-(CH2)dNH-CO-(CH2)eCH3，d＝1～8,e＝1～21；
x＝0～10；
其中，任意C上连接的H可以被取代基所取代；
其中，所述取代基为-NH2、-OH、-COOH、卤素或-Ar中的一种或几种。
在本发明中，-CnH2n+1或-CcH2c+1为烷基，n的取值为1～22。一些实施例中，n的取值为8～20。在另一些实施例中，n为8～18或n为12～18； 在另一些实施例中，n为14～16；在一些实施例中，n为15。c的取值与n的取值可以相同也可以不同，独立的，c的取值为1～22。一些实施例中，c的取值为8～20。在另一些实施例中，c为8～18或c为12～18；在另一些实施例中，c为14～16；在一些实施例中，c为15。所述烷基为直链烷基或支链烷基；在一些实施例中，其为直链烷基。所述烷基为取代或未取代的烷基。所述取代指烷基中任意C上连接的H可以被取代基取代，所述取代烷基的取代基为-NH2、-OH、-COOH、卤素或-Ar中的一种或几种。任意C上的取代基个数为1个或两个。本发明中，-CnH2n+1烷基包括但不限于，甲基(Me，-CH3)，乙基(Et，-CH2CH3)，正丙基(n-Pr，-CH2CH2CH3)，异丙基(i-Pr，-CH(CH3)2)，正丁基(n-Bu，-CH2CH2CH2CH3)，异丁基(i-Bu，-CH2CH(CH3)2)，仲丁基(s-Bu，-CH(CH3)CH2CH3)，叔丁基(t-Bu，-C(CH3)3)，正戊基(-CH2CH2CH2CH2CH3)，2-戊基(-CH(CH3)CH2CH2CH3)，3-戊基(-CH(CH2CH3)2)，2-甲基-2-丁基(-C(CH3)2CH2CH3)，3-甲基-2-丁基(-CH(CH3)CH(CH3)2)，3-甲基-1-丁基(-CH2CH2CH(CH3)2)，2-甲基-1-丁基(-CH2CH(CH3)CH2CH3)，正己基(-CH2CH2CH2CH2CH2CH3)，2-己基(-CH(CH3)CH2CH2CH2CH3)，3-己基(-CH(CH2CH3)(CH2CH2CH3))，2-甲基-2-戊基(-C(CH3)2CH2CH2CH3)，3-甲基-2-戊基(-CH(CH3)CH(CH3)CH2CH3)，4-甲基-2-戊基(-CH(CH3)CH2CH(CH3)2)，3-甲基-3-戊基(-C(CH3)(CH2CH3)2)，2-甲基-3-戊基(-CH(CH2CH3)CH(CH3)2)，2,3-二甲基-2-丁基(-C(CH3)2CH(CH3)2)，3,3-二甲基-2-丁基(-CH(CH3)C(CH3)3)，正庚基，正辛基、正壬基、正癸基、正十一烷基、正十二烷基、正十三烷基、正十四烷基、正十五烷基、正十六烷基、正十七烷基、正十八烷基。
在本发明中，-CnH2n-1或-CcH2c-1指烯基，其中至少一个位置为不饱和状态，即一个C-C为sp2双键。n的取值为2～22。一些实施例中，n的取值为8～20。在另一些实施例中，n为8～18或n为12～18；在另一些实施例中，n为14～16；在一些实施例中，n为15。c的取值与n的取值可以相同也可以不同，独立的，c的取值为2～22。一些实施例中，c的取值为8～20。在另一些实施例中，c为8～18或c为12～18；在另一些实施例中， c为14～16；在一些实施例中，c为15。所述烯基为直链烯基或支链烯基，在一些实施例中，其为直链烯基。所述烯基为取代或未取代的烯基。所述取代烯基中任意C上连接的H可以被取代基取代，所述取代烯基的取代基为-NH2、-OH、-COOH、卤素或-Ar中的一种或几种。任意C上的取代基个数为1个或两个。所述烯基包括基团有“反”“正”或"E""Z"的定位，其中具体的实例包括，但并不限于，乙烯基(-CH＝CH2)，丙烯基，烯丙基(-CH2CH＝CH2)，丁烯基和4-甲基丁烯基、2-戊烯基、己烯基、庚烯基、辛烯基、壬烯基、癸烯基、十一烯基、十二烯基、十三烯基、十四烯基、十五烯基、十六烯基、十七烯基、十八烯基。
在本发明中，-Ar-CnH2n+1指与苯基或芳基相连接的-CnH2n+1，-Ar-CnH2n-1指与苯基或芳基相连接的-CnH2n-1，-Ar-O-CnH2n+1指与-Ar-O-相连接的-CnH2n+1，-Ar-O-CnH2n-1指与-Ar-O-相连接的-CnH2n-1。-Ar-CcH2c+1指与苯基或芳基相连接的-CcH2c+1，-Ar-CcH2c-1指与苯基或芳基相连接的-CcH2c-1，-Ar-O-CcH2c+1指与-Ar-O-相连接的-CcH2c+1，-Ar-O-CcH2c-1指与-Ar-O-相连接的-CcH2c-1。所述-Ar-表示共含有6-14元环的单环，双环，和三环的碳环体系，并且，至少一个环体系是芳香族的，其中每一个环体系包含3-7元环，其中，具有两个连接点与分子其余部分相连。-Ar-可以为亚苯基，萘基和亚蒽基。并且所述-Ar-可以是取代或非取代的。
在本发明中，-CbH2b-或-CdH2d-指亚烷基，b的取值为1～8。一些实施例中，b的取值为1～6。在另一些实施例中，b为1～4；在另一些实施例中，b为2～4；所述亚烷基为直链或支链的亚烷基；d的取值与b的取值可以相同也可以不同，独立的，d的取值为1～8。一些实施例中，d的取值为1～6。在另一些实施例中，d为1～4；在另一些实施例中，d为2～4；所述亚烷基为直链或支链的亚烷基；在一些实施例中，其为直链的亚烷基。所述亚烷基为取代或未取代的亚烷基。所述取代指亚烷基中任意C上连接的H可以被取代基取代，所述取代烷基的取代基为-NH2、-OH、-COOH或-Ar中的一种或几种。任意C上的取代基个数为1个或两个。本发明中，亚烷基包括但不限于，亚甲基(-CH2-)，亚乙基(-CH2CH2-)，正亚丙基(-CH2CH2CH2-)，异亚丙基(-C(CH3)2-)，正亚丁基(-CH2CH2CH2CH2-)，异 亚丁基，仲亚丁基，叔亚丁基，正亚戊基，2-亚戊基，3-亚戊基，2-甲基-2-亚丁基，3-甲基-2-亚丁基，3-甲基-1-亚丁基，2-甲基-1-亚丁基，正亚己基，2-亚己基，3-亚己基，2-甲基-2-亚戊基，3-甲基-2-亚戊基，4-甲基-2-亚戊基，3-甲基-3-亚戊基，2-甲基-3-亚戊基，2,3-二甲基-2-亚丁基，3,3-二甲基-2-亚丁基，正亚庚基，正亚辛基，羧基取代的亚丙基、羧基取代的亚正丁基、羧基取代的亚正戊基、氨基取代的亚丙基、氨基取代的亚正丁基、氨基取代的亚正戊基。
在本发明中，-CbH2b-2-或-CdH2d-2-指亚烯基，其中至少一个位置为不饱和状态，即一个C-C为sp2双键。b的取值为2～8。一些实施例中，b的取值为2～6。在另一些实施例中，b为2～4。d的取值与b的取值可以相同也可以不同，独立的，d的取值为2～8。一些实施例中，d的取值为2～6。在另一些实施例中，d为2～4。所述烯基为直链亚烯基或支链亚烯基，在一些实施例中，其为直链亚烯基。所述亚烯基为取代或未取代的亚烯基。所述取代亚烯基中任意C上连接的H可以被取代基取代，所述取代亚烯基的取代基为-NH2、-OH、-COOH或-Ar中的一种或几种。任意C上的取代基个数为1个或两个。所述亚烯基包括基团有“反”“正”或"E""Z"的定位，其中具体的实例包括，但并不限于，亚乙烯基(-CH＝CH-)，亚丙烯基，亚烯丙基(-CH2CH＝CH-)，亚丁烯基和4-甲基亚丁烯基、2-亚戊烯基、亚己烯基、亚庚烯基、亚辛烯基。
在本发明的实施例中，-CdH2d-NH2、-CdH2d-2-NH2为与氨基(-NH2)相连接的-CdH2d-或-CdH2d-2-；-CdH2d-COOH、-CdH2d-2-COOH为与羧基(-COOH)相连接的-CdH2d-或-CdH2d-2-。
在一些实施例中，
R1选自-H、-CnH2n+1或-CnH2n-1,n＝8～18；
R4选自-CbH2b-、-CbH2b-2-或除去2个羧基的透明质酸,b＝1～10；
R5选自-CcH2c+1、-CcH2c-1或除去1个氨基的壳聚糖,c＝8～18；
R6选自-NH2、-NH-CO-(CH2)eCH3、-CdH2d-NH2、-CdH2d-COOH或-(CH2)dNH-CO-(CH2)eCH3；d＝2～6,e＝8～18。
在一些实施例中，
x＝0～6；
R1为-CnH2n+1，n＝8～18；
R4选自-CH2-、
Figure PCTCN2017071114-appb-000012
Figure PCTCN2017071114-appb-000013
或除去2个羧基的透明质酸；
R5选自-CcH2c+1，c＝12～18；
R6选自-NH2、-(CH2)4NH2、-(CH)2-COOH或-(CH2)4NH-CO-(CH2)14CH3。
在一些实施例中，x＝0；R1为-CnH2n+1，n＝8～18；R6为-CH2-COOH、-(CH2)4NH2、-(CH2)2-COOH或-(CH2)4NH-CO-(CH2)14CH3。
x＝1～6；R1为-CnH2n+1，n＝8～18；R6为-NH2。
本发明提供的铂类配合物如式(1)～(22)，其中y＝7～17：
Figure PCTCN2017071114-appb-000014
Figure PCTCN2017071114-appb-000015
Figure PCTCN2017071114-appb-000016
本发明提供的铂类配合物结构式(1)～(22)中y＝7、8、9、10、11、12、13、14、15、16或17。
本发明提供的铂类配合物由式II化合物或式III化合物与铂类化合物反应制得；所述铂类化合物为顺铂、卡铂、奥沙利铂或奈达铂、依铂、洛铂等。
Figure PCTCN2017071114-appb-000017
R1选自-H、-CnH2n+1、-CnH2m-1、-Ar-CnH2n+1、-Ar-CnH2n-1、-Ar-O-CnH2n+1或-Ar-O-CnH2n-1,n＝1～22；
R2选自-CmH2m-、-CmH2m-2-、-CmH2m-NH-或-CmH2m-2-NH-,m＝1～8；
R3选自-OH、-CaH2a+1、-CaH2a-1、-Ar-CaH2a+1、-Ar-CaH2a-1、-Ar-O-CaH2a+1或-Ar-O-CaH2a-1,a＝1～22；
R4选自-CbH2b-、-CbH2b-2-、-Ar-CbH2b-、-Ar-CbH2b-2-、-Ar-O-CbH2b-、-Ar-O-CbH2b-2-或除去2个羧基的透明质酸,b＝0～22；
R5选自-H、-CcH2c+1、-CcH2c-1、-Ar-CcH2c+1、-Ar-CcH2c-1、-Ar-O-CcH2c+1、-Ar-O-CcH2c-1或除去1个氨基的壳聚糖,c＝0～22；
其中，任意C上连接的H可以被取代基所取代；
其中，所述取代基为-NH2、-OH、-COOH、卤素或-Ar中的一种或几种；
所述R2和R3的结构中至少包括1个-COOH。
在本发明提供的方法中，式II或式III化合物与铂类化合物中Pt的摩尔比不大于30:1。
在一些实施例中，式II或式III化合物与铂类化合物中Pt的摩尔比为(1～8)：1。
在另一些实施例中，式II或式III化合物与铂类化合物中Pt的摩尔比为(1～5)：1。
在另一些实施例中，式II或式III化合物与铂类化合物中Pt的摩尔比为1：1。
制备铂类配合物的反应中采用的溶剂为水性介质。
在本发明提供的实施例中，水性介质为水或甘露醇水溶液或氯化钠水溶液或甘油水溶液或含糖的水溶液或磷酸盐缓冲液等(不仅限于此些)、也可以是含有二甲基甲酰胺(DMF)、二甲基亚砜(DMSO)的上述水溶液。
本发明提供了式II所示结构的两亲性化合物：
Figure PCTCN2017071114-appb-000018
R1选自-H、-CnH2n+1、-CnH2n-1、-Ar-CnH2n+1、-Ar-CnH2n-1、-Ar-O-CnH2n+1或-Ar-O-CnH2n-1,n＝1～22；
R2选自-CmH2m-、-CmH2m-2-、-CmH2m-NH-或-CmH2m-2-NH-,m＝1～8；
R3选自-OH、-CaH2a+1、-CaH2a-1、-Ar-CaH2a+1、-Ar-CaH2a-1、-Ar-O-CaH2a+1或-Ar-O-CaH2a-1,a＝1～22；
其中，任意C上连接的H可以被取代基所取代；
其中，所述取代基为-NH2、-OH、-COOH，卤素或-Ar中的一种或几种；
所述R2和R3的结构中至少包括1个-COOH。
在本发明中，-CnH2n+1或-CaH2a+1为烷基，n的取值为1～22。一些实施例中，n的取值为8～20。在另一些实施例中，n为8～18或n为12～18；在另一些实施例中，n为14～16；在一些实施例中，n为15。a的取值与n的取值可以相同也可以不同，独立的，a的取值为1～22。一些实施例中，a的取值为8～20。在另一些实施例中，a为8～18或a为12～18；在另一些实施例中，a为14～16；在一些实施例中，a为15。所述烷基为直链烷基或支链烷基；在一些实施例中，其为直链烷基。所述烷基为取代或未取代的烷基。所述取代指烷基中任意C上连接的H可以被取代基取代，所述取代烷基的取代基为-NH2、-OH、-COOH或-Ar中的一种或几种。任意C上的取代基个数为1个或两个。本发明中，-CnH2n+1烷基包括但不限于，甲基(Me，-CH3)，乙基(Et，-CH2CH3)，正丙基(n-Pr，-CH2CH2CH3)，异丙基(i-Pr，-CH(CH3)2)，正丁基(n-Bu，-CH2CH2CH2CH3)，异丁基(i-Bu，-CH2CH(CH3)2)，仲丁基(s-Bu，-CH(CH3)CH2CH3)，叔丁基(t-Bu，-C(CH3)3)，正戊基(-CH2CH2CH2CH2CH3)，2-戊基(-CH(CH3)CH2CH2CH3)，3-戊基(-CH(CH2CH3)2)，2-甲基-2-丁基(-C(CH3)2CH2CH3)，3-甲基-2-丁基 (-CH(CH3)CH(CH3)2)，3-甲基-1-丁基(-CH2CH2CH(CH3)2)，2-甲基-1-丁基(-CH2CH(CH3)CH2CH3)，正己基(-CH2CH2CH2CH2CH2CH3)，2-己基(-CH(CH3)CH2CH2CH2CH3)，3-己基(-CH(CH2CH3)(CH2CH2CH3))，2-甲基-2-戊基(-C(CH3)2CH2CH2CH3)，3-甲基-2-戊基(-CH(CH3)CH(CH3)CH2CH3)，4-甲基-2-戊基(-CH(CH3)CH2CH(CH3)2)，3-甲基-3-戊基(-C(CH3)(CH2CH3)2)，2-甲基-3-戊基(-CH(CH2CH3)CH(CH3)2)，2,3-二甲基-2-丁基(-C(CH3)2CH(CH3)2)，3,3-二甲基-2-丁基(-CH(CH3)C(CH3)3)，正庚基，正辛基、正壬基、正癸基、正十一烷基、正十二烷基、正十三烷基、正十四烷基、正十五烷基、正十六烷基、正十七烷基、正十八烷基。
在本发明中，-CnH2n-1或-CaH2a-1指烯基，其中至少一个位置为不饱和状态，即一个C-C为sp2双键。n的取值为2～22。一些实施例中，n的取值为8～20。在另一些实施例中，n为8～18或n为12～18；在另一些实施例中，n为14～16；在一些实施例中，n为15。a的取值与n的取值可以相同也可以不同，独立的，a的取值为2～22。一些实施例中，a的取值为8～20。在另一些实施例中，a为8～18或a为12～18；在另一些实施例中，a为14～16；在一些实施例中，a为15。所述烯基为直链烯基或支链烯基，在一些实施例中，其为直链烯基。所述烯基为取代或未取代的烯基。所述取代烯基中任意C上连接的H可以被取代基取代，所述取代烯基的取代基为-NH2、-OH、-COOH、卤素或-Ar中的一种或几种。任意C上的取代基个数为1个或两个。所述烯基包括基团有“反”“正”或"E""Z"的定位，其中具体的实例包括，但并不限于，乙烯基(-CH＝CH2)，丙烯基，烯丙基(-CH2CH＝CH2)，丁烯基和4-甲基丁烯基、2-戊烯基、己烯基、庚烯基、辛烯基、壬烯基、癸烯基、十一烯基、十二烯基、十三烯基、十四烯基、十五烯基、十六烯基、十七烯基、十八烯基。
在本发明中，-Ar-CnH2n+1指与苯基或芳基相连接的-CnH2n+1，-Ar-CnH2n-1指与苯基或芳基相连接的-CnH2n-1，-Ar-O-CnH2n+1指与-Ar-O-相连接的-CnH2n+1，-Ar-O-CnH2n-1指与-Ar-O-相连接的-CnH2n-1。-Ar-CaH2a+1指与苯基或芳基相连接的-CaH2a+1，-Ar-CaH2a-1指与苯基或芳基相连接的 -CaH2a-1，-Ar-O-CaH2a+1指与-Ar-O-相连接的-CaH2a+1，-Ar-O-CaH2a-1指与-Ar-O-相连接的-CaH2a-1。所述-Ar-表示共含有6-14元环的单环，双环，和三环的碳环体系，并且，至少一个环体系是芳香族的，其中每一个环体系包含3-7元环，其中，具有两个连接点与分子其余部分相连。-Ar-可以为亚苯基，萘基和亚蒽基。并且所述-Ar-可以是取代或非取代的。
在本发明中，-CmH2m-指亚烷基，m的取值为1～8。一些实施例中，m的取值为1～6。在另一些实施例中，m为1～4；在另一些实施例中，m为2～4；所述亚烷基为直链或支链的亚烷基；在一些实施例中，其为直链的亚烷基。所述亚烷基为取代或未取代的亚烷基。所述取代指亚烷基中任意C上连接的H可以被取代基取代，所述取代烷基的取代基为-NH2、-OH、-COOH或-Ar中的一种或几种。任意C上的取代基个数为1个或两个。本发明中，-CmH2m-亚烷基包括但不限于，亚甲基(-CH2-)，亚乙基(-CH2CH2-)，正亚丙基(-CH2CH2CH2-)，异亚丙基(-C(CH3)2-)，正亚丁基(-CH2CH2CH2CH2-)，异亚丁基，仲亚丁基，叔亚丁基，正亚戊基，2-亚戊基，3-亚戊基，2-甲基-2-亚丁基，3-甲基-2-亚丁基，3-甲基-1-亚丁基，2-甲基-1-亚丁基，正亚己基，2-亚己基，3-亚己基，2-甲基-2-亚戊基，3-甲基-2-亚戊基，4-甲基-2-亚戊基，3-甲基-3-亚戊基，2-甲基-3-亚戊基，2,3-二甲基-2-亚丁基，3,3-二甲基-2-亚丁基，正亚庚基，正亚辛基，羧基取代的亚丙基、羧基取代的亚正丁基、羧基取代的亚正戊基、氨基取代的亚丙基、氨基取代的亚正丁基、氨基取代的亚正戊基。
在本发明中，-CmH2m-2-指亚烯基，其中至少一个位置为不饱和状态，即一个C-C为sp2双键。n的取值为2～8。一些实施例中，n的取值为2～6。在另一些实施例中，n为2～4。所述烯基为直链亚烯基或支链亚烯基，在一些实施例中，其为直链亚烯基。所述亚烯基为取代或未取代的亚烯基。所述取代亚烯基中任意C上连接的H可以被取代基取代，所述取代亚烯基的取代基为-NH2、-OH、-COOH或-Ar中的一种或几种。任意C上的取代基个数为1个或两个。所述亚烯基包括基团有“反”“正”或"E""Z"的定位，其中具体的实例包括，但并不限于，亚乙烯基(-CH＝CH-)，亚丙烯基，亚烯丙基(-CH2CH＝CH-)，亚丁烯基和4-甲基亚丁烯基、2-亚戊烯基、 亚己烯基、亚庚烯基、亚辛烯基。
在本发明中，-CmH2m-NH-或-CmH2m-2-NH-指与-NH-相连接的-CmH2m-或-CmH2m-2-。
在本发明的实施例中，两亲性化合物中：
R1选自-CnH2n+1、-CnH2n-1，n＝8～18；
R2选自-CmH2m-、-CmH2m-1-NH-，m＝2～6；
R3选自-OH、-CaH2a+1、-CaH2a-1，a＝8～18。
其中，任意C上连接的H可以被一个或两个取代基所取代；
其中，所述取代基为-NH2或-COOH。
所述R2和R3的结构中至少包括1个-COOH。
在本发明的实施例中，
R1为-CnH2n+1，R2为
Figure PCTCN2017071114-appb-000019
R3为-OH；
或R1为-CnH2n+1；，R2为
Figure PCTCN2017071114-appb-000020
R3为-OH；
或R1为-CnH2n+1，R2为
Figure PCTCN2017071114-appb-000021
R3为-CaH2a+1；
或R1为-CnH2n+1，R2为
Figure PCTCN2017071114-appb-000022
R3为-CaH2a+1；
在本发明的实施例中，其结构为：
Figure PCTCN2017071114-appb-000023
本发明提供的铂类配合物能够与脂质体的膜材良好的结合，从而提高脂质体的包封率和载药量。并且，以本发明提供的化合物制备铂类配合物后，含有羧基具有pH敏感性，在较低的pH值环境中(如肿瘤组织)，羧基趋于去质子化，有利于促进药物在肿瘤组织的释放，提高药物的疗效、降低药物的毒副作用。
本发明还提供了一种铂类配合物的脂质体，包括本发明提供的铂类配合物、脂质膜材和/或稳定剂；所述铂类配合物、脂质膜材的比例为(99：1)～(1：99)。
所述比例为质量比、摩尔比或体积比。
本发明提供的脂质体中，脂质膜材包括磷脂和/或胆固醇的组合物，膜材中磷脂的质量份为1份～100份，胆固醇的质量份为0份～60份；
在本发明的实施例中，磷脂为蛋黄卵磷脂、大豆卵磷脂、氢化大豆磷脂、磷脂酰丝氨酸、磷脂酰肌醇、磷脂酰乙醇胺、磷脂酰甘油、二月桂酰卵磷脂、二肉豆蔻酰基卵磷脂、二棕榈酰基卵磷脂或二硬脂酰基卵磷脂、二油酰基磷脂酰胆碱、二月桂酰磷脂酰甘油、二肉豆蔻酰基磷脂酰甘油、二棕榈酰基磷脂酰甘油或二硬脂酰基磷脂酰甘油、二油酰基磷脂酰甘油、磷酸酰甘油、二芥酰磷脂酰甘油、PEG化磷脂等中的任一种或两者以上的组合物。
本发明提供的脂质体中，或也还包括长循环材料，本发明采用的长循环材料为MPEG2000-DSPE，MPEG5000-DSPE、MPEG2000-DMPE，MPEG5000-DMPE，MPEG2000-DPPE，MPEG5000-DPPE等。
本发明提供的脂质体中长循环材料的质量分数为0～50％。
本发明提供的脂质体中还可包括抗氧化剂；本发明采用的抗氧化剂为L-半胱氨酸、抗坏血酸、dl-ɑ-生育酚、亚硫酸钠、焦亚硫酸钠或无水亚硫酸氢钠中任一种或两者以上的组合物。
本发明提供的脂质体中，抗氧化剂的质量分数不高于20％。
本发明提供的脂质体中，还可加入等渗调节剂，所述等渗调节剂为甘露醇、山梨醇、葡萄糖、蔗糖或氯化钠中任一种或两者以上的组合物。
本发明提供脂质体的制备方法可以采用薄膜分散法、逆向蒸发法或 复乳法等。
在一些实施例中，脂质体的制备方法为：取膜材、抗氧化剂、长循环材料置于圆底烧瓶中，加入适量的有机溶剂溶解，加入本发明提供的铂类配合物的水性溶液，机械或超声均化形成W/O乳剂，减压旋转蒸发除去有机溶剂，在容器壁上形成薄膜，注入水化介质，使得膜材水化1h～2h，过脂质体挤压器或匀化得到粒径均一新型铂类脂质体，长循环材料也可以在脂质体形成后加入，搅拌，修饰于脂质体表面。
在另一些实施例中，脂质体的制备方法为：取膜材、抗氧化剂与本发明提供的铂类配合和长循环材料置于圆底烧瓶中，加入适量的有机溶剂溶解，减压旋转蒸发除去有机溶剂，在容器壁上形成薄膜，注入水化介质，使得膜材水化1h～2h，过脂质体挤压器或匀化得到新型铂类脂质体。
在另一些实施例中，脂质体的制备方法为：取膜材、抗氧化剂和长循环材料置于圆底烧瓶中，加入适量的有机溶剂溶解，加入本发明提供的铂类配合物，超声或机械均化形成W/O乳剂，减压旋转蒸发除去部分有机溶剂，注入水化介质，机械搅拌形成W/O/W后，减压旋转蒸发除去有机溶剂，过脂质体挤压器或匀化得到新型铂类脂质体。
本发明还提供了一种治疗癌症的药物，包括本发明提供的脂质体及制剂。
本发明提供的药物可用于治疗的癌症，如乳腺癌、非小细胞肺癌、睾丸癌、胰腺癌、直肠癌、肝癌，卵巢癌或头颈部癌。
本发明提供的药物的剂型为注射剂(混悬型或注射用冻干粉末)、乳剂、亚微乳剂等。
试验表明，本发明提供的脂质体能够显著提高铂类化合物的血药浓度，延长药物在体内的滞留时间，并且可以使铂类药物浓集于癌变器官，不仅可以减少用药剂量，增加疗效，而且可以降低药物的毒副作用，适用于多种肿瘤疾病的治疗。
本发明提供了式II结构的化合物，该化合物具有良好的生物相容性，其具有的羧基与酰胺基团能够与铂类药物配合，其与铂类药物形成的不稳定的配位键能够增加药物在体内的水合速度从而提高药效。同时，以本发 明提供的化合物制得的铂类配合物能够与脂质体的膜材良好的结合，从而提高脂质体的包封率和载药量。并且，以本发明提供的化合物制备铂类配合物后，含有羧基具有pH敏感性，在较低的pH值环境中(如肿瘤组织)，羧基趋于去质子化，有利于促进药物在肿瘤组织的释放，更进一步提高药物的疗效、降低其毒副作用。实验表明，本发明提供的脂质体能够显著降低药物的毒副作用并提高药效。
本发明采用的试剂或仪器皆为普通市售品，皆可于市场购得。
下面结合实施例，进一步阐述本发明：
实施例1～8两亲性化合物及其制备：
Figure PCTCN2017071114-appb-000024
其制备方法为：
1.实施例1～5
取1.5mmol的脂肪胺(如十八胺、十六胺、十四胺或十二胺等)加入20mL氯仿溶解后，另取3.3mmol酸酐(如马来酸酐或丁二酸酐)溶于20mL氯仿中，溶解后将脂肪胺的氯仿溶液加至酸酐溶液中，搅拌30min，减压过滤，将得到的产物用氯仿，丙酮依次洗涤后,干燥(投料摩尔比为1:2)，即得上述(实施例1-实施例5)两亲性化合物。对制得两亲性化合物进行核磁结构表征，结果如图1-a～1-e。
2.实施例6～8
取1.5mmol的十八胺20mL二氯甲烷溶解后，另取3.3mmol酸(如醋酸酐、丙酸酐、甲基丁二酸酐)20mL二氯甲烷溶解,将脂肪胺的二氯甲烷溶液加至酸酐溶液中，搅拌30min，减压过滤，将得到的产物用二氯甲烷，丙酮依次洗涤后,干燥(投料摩尔比为1:2)，即得上述(实施例6～实施例8)两亲性化合物。
实施例9两亲性化合物及其制备：
Figure PCTCN2017071114-appb-000025
制备方法为：
由棕榈酰氯与谷氨酸反应合成，具体的反应条件如下：
将10mmol的谷氨酸混悬于10ml DMF中，置于45℃油浴中搅拌，滴加12mmol棕榈酰氯，待到谷氨酸全部溶解，加入200ul三乙胺搅拌1-2h，加入100ml蒸馏水，用1M HCl调节pH至1-2，减压抽滤，用水洗涤。将水洗后的材料于50℃烘箱烘干，取出，用石油醚：丙酮：乙酸乙酯(100：1：1)的混合溶剂加热洗涤三次，离心，取出，减压干燥。
对制得化合物进行核磁结构表征，结果如图1-f。
实施例10两亲性化合物及其制备：
Figure PCTCN2017071114-appb-000026
其制备方法为：
将10mmol的赖氨酸混悬于10ml DMF中，置于45℃油浴中搅拌，滴加24mmol棕榈酰氯，待到赖氨酸全部溶解，加入200ul三乙胺搅拌1-2h，加入100ml蒸馏水，用1M HCl调节pH至1-2，减压抽滤，用水洗涤。
将水洗后的材料于50℃烘箱烘干，取出，用石油醚：丙酮：乙酸乙酯(100：1：1)的混合溶剂加热洗涤三次，离心，取出，减压干燥。对制得化合物进行核磁结构表征，结果如图1-g。
实施例11两亲性化合物及其制备：
Figure PCTCN2017071114-appb-000027
其制备方法为：
将10mmol的赖氨酸混悬于10ml DMF中，置于45℃油浴中搅拌，滴加12mmol棕榈酰氯，待到赖氨酸全部溶解，加入200ul三乙胺搅拌1-2h，加入100ml蒸馏水，用1M HCl调节pH至1-2，减压抽虑，用水洗涤。将水洗后的材料于50℃烘箱烘干，取出，用石油醚：丙酮：乙酸乙酯(100：1：1)的混合溶剂加热洗涤三次，离心，取出沉淀，减压干燥。
对制得化合物进行核磁结构表征，结果如图1-h。
实施例12～27铂类配合物的制备
表1实施例12～27
Figure PCTCN2017071114-appb-000028
Figure PCTCN2017071114-appb-000029
Figure PCTCN2017071114-appb-000030
制备方法：将两亲性化合物加入水中在一定条件下使其搅拌溶解，加入铂类化合物(两亲性化合物与铂类化合物的摩尔比1:1-10：1)，加热搅拌12h，反应得到铂类配合物，经干燥得到铂类配合物的固体粉末。其红外检测光谱如图2-1a～图2-9b。
顺铂与两亲性化合物形成铂类配合物后，红外光谱图发生变化，原两亲性化合物中羧基的νOH峰消失(3076cm-1),酯键中的νC＝O由1720cm-1偏移至1630cm-1-1640cm-1左右,酰胺键中的νC＝O由1643cm-1偏移至1530cm-1-1570cm-1左右并且出现顺铂的νC-N在1420-1440cm-1左右。
两亲性化合物与奥沙利铂形成配合物，原两亲性化合物中羧基的νOH峰消失(3076cm-1，920cm-1)，酯键中的νC＝O由1720cm-1偏移至1650cm-1-1700cm-1左右，酰胺键中的νC＝O由1643cm-1偏移至1530 cm-1-1570cm-1左右并且出现奥沙利铂的环己二胺νs(N-H)在3220cm-1及环己二胺ν(C-N)在1100-1200cm-1。
实施例28～37逆向蒸发法制备脂质体
表2实施例28～37
Figure PCTCN2017071114-appb-000031
取配方量的膜材、抗氧化剂置于圆底烧瓶中，加入适量的有机溶剂溶解(有机溶剂选用：氯仿或水饱和乙醚或二氯甲烷、甲醇、乙醇或其中至少两种溶剂的混合溶剂)，溶解，加入处方量的铂类配合物水溶液，(有机溶剂：水相体积比＝3:1～6:1)，机械或超声均化形成W/O乳剂，减压旋转蒸发除去有机溶剂，在容器壁上形成薄膜，注入水化介质(生理盐水或5％葡萄糖或注射用水或甘露醇水溶液)，40℃(或55℃)使得膜材水化1h～2h，过脂质体挤压器或微射流匀化得到铂类配合物脂质体，加入长循环材料搅拌室温搅拌1h，即得。
实施例38薄膜分散法制备脂质体
表3实施例38
Figure PCTCN2017071114-appb-000032
取配方量的膜材和抗氧化剂、长循环材料、加入配方量的铂类配合物置于圆底烧瓶中，加入适量的氯仿溶解，减压旋转蒸发除去有机溶剂，在容器壁上形成薄膜，注入水化介质(注射用水)，40～55℃使得膜材水化1～2h，过脂质体挤压器匀化得到新型铂类脂质体。
实施例39复乳法制备脂质体
表4实施例39
Figure PCTCN2017071114-appb-000033
取配方量的膜材、抗氧化剂置于圆底烧瓶中，加入适量的乙醚，溶解，加入配方量的铂类化合物水溶液(有机相与水相体积比3:1)，超声或机械匀化形成W/O乳剂，减压旋转蒸发除去部分有机溶剂，注入注射 用水，机械搅拌形成W/O/W后，减压旋转蒸发除去有机溶剂，过脂质体挤压器、微射流匀化匀化分散得到新型铂类脂质体。
实施例40脂质体质量考察
用电镜观察本发明制得的含铂类配合物脂质体的形态，用Zetasizer Nano ZS90激光粒度测定仪测定脂质体的粒径及分布。以《中国药典》提供的方法计算本发明制得脂质体的包封率。结果如表5：
表5脂质体粒度及包封率考察结果
实施例序号 平均粒径(nm) 包封率(％)
28 130.1 94.1
29 132.2 92.7
30 112.6 92.8
31 101.9 93.1
32 113.7 92.7
33 98.45 96.3
34 108.7 95.3
35 112.1 94.4
36 102.6 97.2
37 106.1 92.9
图3、图4为实施例31制得的脂质体的形态和粒度图谱，其他实施例制得脂质体的检测情况与此相似。
结果表明，本发明提供的新型铂类配合物脂质体制剂的粒径在90～150nm之间，包封率为92～98％之间，载药量在8-30％之间。与现有文献及专利已报道的脂质体不同，该铂类脂质体具有包封率高，稳定性好及抗癌活性强的特点。
实施例41脂质体的药效
1、癌细胞存活率及IC50
以市售注射用顺铂为参比和实施例31、33、36制得的脂质体为实验 材料，采用胰酶消化对数期MCF-7，A549，HEPG2细胞，终止后离心收集，制成细胞悬液，细胞计数调整其浓度至5-10×104/mL。将细胞悬液制备好后，轻轻混匀，每孔加入100μL,将接种好的细胞放入培养箱中培养24h，分别加入1μmol/L,2μmol/L,4μmol/L,8μmol/L,16μmol/L,32μmol/L现有顺铂注射剂及实施例20～31制得的脂质体溶液，采用5％CO2，37℃孵育48小时。每孔加入10μLMTT溶液(5mg/mL，0.5％MTT)，继续培养4h后终止培养，每孔加入150μL二甲基亚砜，置摇床上低速振荡10min，使结晶物充分溶解。在酶联免疫检测仪OD 570nm处测量各孔的吸光值。通过公式计算各制剂的IC50值结果见表6。
表6顺铂及实施例31、33和36制得的脂质体对不同肿瘤细胞的IC50(μM)
实施例序号 MCF-7 A549 HEPG2
注射用顺铂 16.9 8.92 7.19
实施例31 3.02 1.67 1.26
实施例33 4.11 2.19 3.28
实施例36 4.22 3.97 1.19
结果显示，本发明提供的铂类配合物的脂质体能够显著(p<0.01)降低对肿瘤细胞的IC50值，药效明显提高。
2、对肿瘤的抑制率
以生理盐水、注射用顺铂、实施例31、33、36制得的脂质体为实验材料，取SPF级KM小鼠，SPF级，均为雄性，6-8周龄，体重18～22g。取对数增长的肝癌细胞H22，将单细胞浓度调至为(1-5)×106cell/mL。将H22细胞悬液接种于小鼠的右侧腋窝皮下，每只0.2mL，建立肝癌H22瘤株腋下接种模型。将荷瘤小鼠随机分为5组，每组10只，分别为生理盐水组，注射用顺铂对照组，实验组(分别给予本发明实施例31、33、36提供的铂类配合物脂质体)，组内标号，各组尾静脉注射生理盐水，顺铂对照组，实验组，每隔三天给药一次，给药两次，给药剂量分别为2.5mg/kg。给药后，每日观察小鼠的存活状态，给药后第7天处死小鼠，取出肿瘤，生理盐水洗净表面，滤纸吸干，称重，计算抑瘤率，计算方法如下：
肿瘤抑制率(％)＝(对照组的平均瘤重-受试组的平均瘤重)÷对照组的平均瘤重×100
实验结果显示，给药组的肿瘤生长均比对照组的肿瘤生长缓慢，实验组与顺铂对照组相比较，能够显著的抑制肿瘤的生长，同时可以显著的延长荷瘤小鼠的生存时间。肿瘤体积及相关参数分别见表7：
表7荷肝癌H22的KM小鼠尾静脉注射2.5mg/kg顺铂对照组和实验组的肿瘤体积及肿瘤抑制率
实施例序号 肿瘤重量(g) 抑制率(％)
生理盐水 1.220  
顺铂注射剂 0.661 45.8
实施例31 0.351 71.2
实施例33 0.338 72.3
实施例36 0.403 67.0
结果表明，与顺铂注射液相比，本发明提供的脂质体能够更显著的抑制肿瘤生长，其效果与顺铂注射液相比具有显著差异(p<0.01)。
实施例42脂质体安全性
1、存活状态、体重
将KM小鼠随机分为组，每组5只，分别为空白对照组、注射用顺铂对照组、实施例31、33、36制得的脂质体,组内标号，各组尾静脉注射分别为生理盐水、市售顺铂对照组，实施例31、33、36制得的脂质体,给药1次，给药剂量分别为10mg/kg、20mg/kg，给药后，每日观察小鼠的存活状态，称量其体重，记录小鼠存活率。
结果见图5和图6，结果表明与顺铂对照组相比，实施例31、33、36制得的脂质体小鼠体重下降明显减轻，存活率显著提高，表明顺铂配合物脂质体系统毒性显著降低。
实施例43脂质体安全性
2、肾毒性
将KM小鼠随机分为5组，每组10只，空白对照组、分别为实施例 31、33、36制得的脂质体,市售注射用顺铂对照组，组内标号，各组尾静脉注射生理盐水、实施例31、33、36制得的脂质体溶液,顺铂注射液，给药1次，给药剂量分别为10mg/kg。给药后，每日观察小鼠的存活状态，给药后第7天处死小鼠，并分别测定小鼠尿素氮和肌酐值，结果见表8：
表8小鼠尿素氮水平和肌酐水平
  肌酐值(μmol·L-1) 尿素氮(mmol·L-1)
生理盐水 69.66±16.38 8.77±1.68
注射用顺铂 134.96±26.24 13.49±0.85
实施例31 74.25±21.22 8.65±0.90
实施例33 77.82±27.47 7.95±1.23
实施例36 68.82±15.47 8.85±1.19
实验结果表明与顺铂注射剂相比，本发明所制备的顺铂配合物脂质体能够显著降低顺铂的肾毒性。
以上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。

Claims (13)

  1. 式I-a～I-d所示结构的铂类配合物：
    Figure PCTCN2017071114-appb-100001
    R1选自-H、-CnH2n+1、-CnH2n-1、-Ar-CnH2n+1、-Ar-CnH2n-1、-Ar-O-CnH2n+1或-Ar-O-CnH2n-1,n＝1～22；
    R4选自-CbH2b-、-CbH2b-2-、-Ar-CbH2b-、-Ar-CbH2b-2-、-Ar-O-CbH2b-、-Ar-O-CbH2b-2-或除去2个羧基的透明质酸,b＝0～22；
    R5选自-H、-CcH2c+1、-CcH2c-1、-Ar-CcH2c+1、-Ar-CcH2c-1、-Ar-O-CcH2c+1、-Ar-O-CcH2c-1或除去1个氨基的壳聚糖,c＝1～22；
    R6选自-NH2、-NH-CO-(CH2)eCH3、-CdH2d-NH2、-CdH2d-2-NH2、-CdH2d-COOH、-CdH2d-2-COOH或-CdH2d-NH-CO-(CH2)eCH3，d＝1～8,e＝1～21；
    x＝0～10；
    其中，任意C上连接的H可以被取代基所取代；
    其中，所述取代基为-NH2、-OH、-COOH、卤素或-Ar中的一种或几种。
  2. 根据权利要求1所述的铂类配合物，其特征在于，
    x＝0～10；
    R1选自-H、-CnH2n+1或-CnH2n-1,n＝8～20；
    R4选自-CbH2b-、-CbH2b-2-或除去2个羧基的透明质酸,b＝1～10；
    R5选自-CcH2c+1、-CcH2c-1或除去1个氨基的壳聚糖,c＝8～20；
    R6选自-NH2、-NH-CO-(CH2)eCH3、-CdH2d-NH2、-CdH2d-COOH或-(CH2)dNH-CO-(CH2)eCH3；d＝1～6,e＝8～20。
  3. 根据权利要求1所述的铂类配合物，其特征在于，
    x＝0；R1为-CnH2n+1，n＝8～18；R6为-(CH2)2-COOH、-CH2-COOH、-(CH2)4NH2、或-(CH2)4NH-CO-(CH2)14CH3。
  4. 根据权利要求1所述的铂类配合物，其特征在于，x＝1～6；R1为-CnH2n+1，n＝8～18，R6为-NH2。
  5. 根据权利要求1所述的铂类配合物，其特征在于，
    R4选自-CH2-、
    Figure PCTCN2017071114-appb-100002
    Figure PCTCN2017071114-appb-100003
    或除2个羧基的透明质酸；
    R5选自-CcH2c+1，c＝12～18。
  6. 根据权利要求1所述的铂类配合物，其特征在于，结构如式(1)～(22)：其中y＝7～17：
    Figure PCTCN2017071114-appb-100004
    Figure PCTCN2017071114-appb-100005
    Figure PCTCN2017071114-appb-100006
  7. 式II所示结构的两亲性化合物：
    Figure PCTCN2017071114-appb-100007
    R1选自-H、-CnH2n+1、-CnH2m-1、-Ar-CnH2n+1、-Ar-CnH2n-1、-Ar-O-CnH2n+1或-Ar-O-CnH2n-1,n＝1～22；
    R2选自-CmH2m-、-CmH2m-2-、-CmH2m-NH-或-CmH2m-2-NH-,m＝1～8；
    R3选自-OH、-CaH2a+1、-CaH2a-1、-Ar-CaH2a+1、-Ar-CaH2a-1、-Ar-O-CaH2a+1或-Ar-O-CaH2a-1,a＝1～22；
    其中，任意C上连接的H可以被取代基所取代；
    其中，所述取代基为-NH2、-OH、-COOH、卤素或-Ar中的一种或几种；
    所述R2和R3的结构中至少包括1个-COOH。
  8. 根据权利要求7所述的两亲性化合物，其特征在于，
    R1为-CnH2n+1，R2为
    Figure PCTCN2017071114-appb-100008
    R3为-OH；
    或R1为-CnH2n+1，R2为
    Figure PCTCN2017071114-appb-100009
    R3为-OH；
    或R1为-CnH2n+1，R2为
    Figure PCTCN2017071114-appb-100010
    R3为-CaH2a+1；
    或R1为-CnH2n+1，R2为
    Figure PCTCN2017071114-appb-100011
    R3为-CaH2a+1；
    n＝8～18；m＝2～6；a＝8～18。
  9. 根据权利要求7所述的两亲性化合物，其特征在于，其结构如式II-a～式II-b所示：
    Figure PCTCN2017071114-appb-100012
  10. 一种铂类配合物的脂质体，其特征在于，包括：权利要求1～6任一项所述的铂类配合物、脂质膜材和/或稳定剂；所述铂类配合物、脂质膜材比例为(99：1)～(1：99)；所述比例为质量比、摩尔比或体积比。
  11. 根据权利要求10所述的脂质体，其特征在于，所述脂质膜材包括磷脂和/或胆固醇的组合物，膜材中磷脂的质量份为1份～100份，胆固醇的质量份为0份～60份。
  12. 根据权利要求10所述的脂质体，其特征在于，所述磷脂为蛋黄卵磷脂、大豆卵磷脂、氢化大豆磷脂、磷脂酰丝氨酸、磷脂酰肌醇、磷脂酰乙醇胺、磷脂酰甘油、二月桂酰卵磷脂、二肉豆蔻酰基卵磷脂、二棕榈酰基卵磷脂或二硬脂酰基卵磷脂、二油酰基磷脂酰胆碱、二月桂酰磷脂酰甘油、二肉豆蔻酰基磷脂酰甘油、二棕榈酰基磷脂酰甘油或二硬脂酰基磷脂酰甘油、二油酰基磷脂酰甘油、磷酸酰甘油、二芥酰磷脂酰甘油、PEG化磷脂等中的任一种或两者以上的组合物。
  13. 一种治疗癌症的药物，其特征在于，包括权利要求10～12任一项所述的脂质体。
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