Platinum complex, liposome thereof and application thereof
阅读说明：本技术 一种铂类配合物及其脂质体和应用 (Platinum complex, liposome thereof and application thereof ) 是由 杨丽 李青岭 杨阔 于 2020-06-17 设计创作，主要内容包括：本发明涉及医药领域,尤其涉及一类铂类配合物及其脂质体和应用。本发明提供了式Ⅰ-a～Ⅰ-d所示结构的铂类配合物：其中,R-(1)-R-(4)、m如权利要求和说明书所述。本发明提供的铂类配合物不仅能与脂质体膜材良好的结合,提高脂质体的包封率和载药量,而且含有羧基,具有pH敏感性,在肿瘤组织易发生去质子化而有利于药物的释放。实验表明,本发明提供的脂质体能够显著降低药物毒副作用并保持疗效。(The invention relates to the field of medicines, in particular to a platinum complex, a liposome thereof and application thereof. The invention provides a platinum complex with a structure shown in formulas I-a-I-d: wherein R is 1 ‑R 4 M is as described in the claims and specification. The platinum complex provided by the invention can be well combined with a liposome membrane material, the entrapment rate and the drug-loading capacity of the liposome are improved, and the platinum complex contains carboxyl, has pH sensitivity, is easy to deprotonate in tumor tissues and is beneficial to the release of drugs. Experiments show that the liposome provided by the invention can obviously reduce the toxic and side effects of the medicine and keep the curative effect.)
1. Platinum complexes of the structures shown in formulas I-a-I-d:
wherein R is1Is selected from-CnH2n-、-CnH2n-2-、-Ar-CnH2n-、Ar-CnH2n-2-、-Ar-O-CnH2n-、Ar-O-CnH2n-2-，n＝0～22；
R2Is selected from-H, -CbH2b-1、-CbH2b+1、-Ar-CbH2b-1、-Ar-CbH2b+1、-Ar-O-CbH2b-1or-Ar-O-CbH2b+1，b＝1～22；
R3Is selected from-NH2、-(CH2)c-NHCO-(CH2)dCH3、-OH、-O-CO-(CH2)cCH3、-CdH2d-OH、-CdH2d-2-OH、-CdH2d-COOH、-CdH2d-2-COOH、-CdH2d-O-CO-(CH2)c-CH3，c＝1～21，d＝1～8；
R4Is selected from-H, -CeH2e-1、-CeH2e+1、-Ar-CeH2e-1、-Ar-CeH2e+1、-Ar-O-CeH2e-1or-Ar-O-CeH2e+1，e＝1～22；
m＝0～10；
Wherein any H attached to C may be substituted with a substituent;
wherein, the substituent is-OH, -NH2-COOH, halogen or-Ar;
ar is a 6-14 membered monocyclic, bicyclic, or tricyclic carbocyclic ring system, and at least one ring system is aromatic; preferably phenylene, naphthyl, anthracenylene.
2. The platinum-based complex according to claim 1, wherein:
m＝0～10；
R1is selected from-CnH2n-or-CnH2n-2，n＝1～10；
R2Is selected from-CbH2b-1or-CbH2b+1，b＝8～20；
R3Is selected from-NH2、-(CH2)c-NHCO-(CH2)dCH3、-OH、-O-CO-(CH2)cCH3、-CdH2d-OH、-CdH2d-COOH、-CdH2d-2-COOH、-CdH2d-O-CO-(CH2)c-CH3、，c＝8～20，d＝1～6。
R4Is selected from-H, -CeH2e-1or-CeH2e+1，e＝8～20。
3. The platinum-based complex according to claim 1, wherein:
m＝0；R1is-CnH2n-or-CnH2n-2-，n＝8～18。
4. The platinum-based complex according to claim 1, wherein:
R3is- (CH)2)2-COOH、-CH2-COOH、-(CH2)c-CO-(CH2)c-CH3，c＝8～18。
5. The platinum-based complex according to claim 1, wherein: m is 1-6; r2is-CbH2b+1，b＝8～18。
6. The platinum-based complex according to claim 1, wherein:
R1is selected from-CH2-、
R2Is selected from-CbH2b+1，b＝12～18。
7. The platinum complex according to claim 1, which has the structural formula (1) to (18):
wherein x is 7-17:
8. a liposome of a platinum complex, which is characterized by comprising the platinum complex of any one of claims 1 to 7 and a lipid membrane material; the ratio of the platinum complex to the lipid membrane material is (99:1) - (1: 99); the ratio is a mass ratio or a molar ratio.
9. The liposome of claim 8, wherein the lipid membrane material comprises a composition of phospholipids or/and cholesterol, wherein the mass portion of the phospholipids in the membrane material is 1-100 parts, and the mass portion of the cholesterol is 0-60 parts.
10. The liposome of claim 9, wherein the phospholipid comprises one or a combination of two or more of lecithin, soybean phospholipid, hydrogenated soybean phospholipid, phosphatidylinositol, phosphatidylglycerol, dipalmitoylphosphatidylcholine, dimyristoylphosphatidylcholine, distearoylphosphatidylcholine, dilauroylphosphatidic acid, dilauroylphosphatidylglycerol, dimyristoylphosphatidylglycerol, palmitoylphosphatidylglycerol, distearoylphosphatidylglycerol, phosphatidylserine, dilauroylphosphatidylethanolamine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine, distearoylphosphatidylethanolamine, pegylated phospholipid, and the like.
11. Use of a platinum complex according to any one of claims 1 to 7 or a liposome according to claim 8 for the manufacture of an anti-cancer medicament.
Technical Field
The invention relates to the field of medicines, and particularly relates to a platinum complex, a liposome thereof and application thereof.
Background
The platinum compound has some unique and excellent physicochemical properties, plays an important role in high and new technology, is known as vitamin of modern industry, so the research and the application of the platinum compound are rapidly developed since the Rosenborg first discovered that the platinum compound has anti-tumor activity in the 60 s of the 20 th century. Cisplatin, carboplatin, nedaplatin, eptaplatin, cycloplatin, oxaliplatin, leplatin, heptaplatin, and the like have been successfully developed at present, and new platinum complexes are continuously synthesized and applied to clinical tests. Although all the platinum anti-tumor drugs have certain anti-tumor activity, the drugs with the curative effect superior to that of cisplatin are only discovered. According to statistics, in the anti-tumor chemotherapy treatment scheme in China, the scheme mainly comprising cisplatin or with cisplatin participating in compatibility accounts for 70-80% of all chemotherapy schemes. The platinum antineoplastic drugs are broad-spectrum anticancer drugs, have the advantages of strong action, high activity, no cross drug resistance, easy compatibility with other antineoplastic drugs and the like, but have serious toxic and side effects (including nephrotoxicity, neurotoxicity, ototoxicity, gastrointestinal toxicity and the like) and drug resistance which greatly limit large clinical dose and long-term use.
Since the non-specificity caused by broad-spectrum antitumor activity is the main reason of toxic and side effects of platinum drugs, finding platinum antitumor drugs with low toxic and side effects on the basis of retaining antitumor activity has become the current main research direction. Therefore, various countries of pharmaceutical workers are engaged in the research on the synthesis of new platinum compounds and targeted drug delivery systems (such as liposomes, nanoparticles, micelles, vesicles, etc.), and it is expected that high-activity and low-toxicity antitumor drugs can be obtained.
At present, various antitumor drugs developed by carrying platinum drugs on carriers have been published. Inhaled lipid-complexed cisplatin (ILC) is a lung-Inhaled cisplatin liposome compound developed by Intel Gene technology Limited in China [ Patent, USA:20140065205A1, 2014 ], is a sustained-release nano-liposome-coated cisplatin novel dosage form capable of being administrated by aerosol inhalation, shows excellent safety and good treatment effect in a stage I lung cancer test and a clinical Ib/IIa osteosarcoma lung metastasis test which are completed in Europe and America, and the lung lesion tumor of a patient is continuously reduced along with the increase of a treatment period. In addition, researchers also prepare antitumor drugs into liposomes, and expect that the purposes of reducing toxicity and improving curative effect are achieved by changing the distribution of the drugs in vivo.
However, platinum-based antitumor drugs using liposomes as carriers still face many challenges. Cisplatin asFor example, SPI-77 is a cisplatin long-circulating liposome developed by Alza corporation using passive drug loading, and has very good pharmacokinetic properties, such as t in mice1/2About 16h, 66 times more than cisplatin, prolongs its circulation time in blood. Furthermore, compared to cisplatin, its plasma AUC increased sixteenfold, Cmax increased three-fold, platinum volume in the kidney decreased four-fold, and tumor AUC increased 28-fold [ Cancer Chemother Pharmacol, 1999, 43 (1): 1-7 ] however, SPI-77 is indicated in the treatment of inoperable head and neck cancer [ Ann Oncol, 2001, 12 (4): 493496 ], advanced non-small cell lung Cancer [ Br J Cancer, 2006, 95 (7): 822-828 ] and recurrent ovarian cancer sensitive to platinum [ Anticancer Res, 2010, 30 (2): 541-545 ] lack of anti-tumor activity in phase II clinical trials. It is thought that hydrophilic drugs such as cisplatin cannot pass through phospholipid membranes smoothly, and only when liposomes release the entrapped drug in tumor tissues, the drugs can have antitumor activity. Lipoplatin is another cisplatin long-circulating liposome developed by Regulon corporation, which is a liposome prepared by forming a reverse micelle of cisplatin and dipalmitoyl phosphatidylglycerol (DPPG) and then encapsulating the reverse micelle in a lipid membrane. Clinical research shows that Lipoplatin can reduce nephrotoxicity, neurotoxicity, ototoxicity and bone marrow toxicity, and has curative effect similar to that of cisplatin or even superior to that of cisplatin. Its accumulation in solid tumors is 10-200 times higher than in normal tissues [ Anticancer Res, 2005, 25 (4): 3031-. Therefore, how to prepare platinum antineoplastic drugs with high drug loading, high activity and low toxicity remains a great challenge to pharmaceutical workers at present.
Disclosure of Invention
The invention aims to provide a platinum complex and a liposome thereof. The platinum complex and the liposome prepared from the platinum complex have high entrapment rate and drug-loading rate, remarkably reduce toxicity, especially renal toxicity, have simple and controllable preparation process and are beneficial to industrial expanded production.
The invention provides platinum complexes with structures shown in formulas I-a to I-b:
R1is selected from-CnH2n-、-CnH2n-2-、-Ar-CnH2n-、Ar-CnH2n-2-、-Ar-O-CnH2n-、Ar-O-CnH2n-2-，n＝0～22；
Wherein Ar is a 6-14 membered ring monocyclic, bicyclic, or tricyclic carbocyclic ring system, and at least one ring system is aromatic; preferably phenylene, naphthyl, anthracenylene;
R2is selected from-H, -CbH2b-1、-CbH2b+1、-Ar-CbH2b-1、-Ar-CbH2b+1、-Ar-O-CbH2b-1or-Ar-O-CbH2b+1，b＝1～22；
Wherein Ar is a 6-14 membered ring monocyclic, bicyclic, or tricyclic carbocyclic ring system, and at least one ring system is aromatic; preferably phenylene, naphthyl, anthracenylene;
wherein any H attached to C may be substituted with a substituent;
wherein, the substituent is-OH, -NH2-COOH, halogen or-Ar; ar is as described above;
in an embodiment of the present invention, it is,
R1is selected from-CnH2n-or-CnH2n-2N is 1-10; n is preferably 1-4;
R2is selected from-CbH2b-1or-CbH2b+1，b＝8～18；
In an embodiment of the present invention, it is,
R1is selected from-CH2-、
R2Is selected from-CbH2b-1or-CbH2b+1，b＝12～18。
In some embodiments of the present invention, the,
R1is composed of
R2Is- (CH)2)17CH3。
The invention also provides platinum complexes with structures shown in formulas I-c-I-d:
R3is selected from-NH2、-(CH2)c-NHCO-(CH2)dCH3、-OH、-O-CO-(CH2)cCH3、-CdH2d-OH、-CdH2d-2-OH、-CdH2d-COOH、-CdH2d-2-COOH、-CdH2d-O-CO-(CH2)c-CH3、，c＝1～21，d＝1～8；
R4Is selected from-H, -CeH2e-1、-CeH2e+1、-Ar-CeH2e-1、-Ar-CeH2e+1、-Ar-O-CeH2e-1or-Ar-O-CeH2e+1，e＝1～22；
Wherein the content of the first and second substances,
ar is a 6-14 membered monocyclic, bicyclic, or tricyclic carbocyclic ring system, and at least one ring system is aromatic; preferably phenylene, naphthyl, anthracenylene;
m＝0～10；
wherein any H attached to C may be substituted with a substituent;
wherein, the substituent is-OH, -NH2-COOH, halogen or-Ar.
In an embodiment of the present invention, it is,
m＝0～8；
R3is selected from-NH2、-(CH2)c-NHCO-(CH2)dCH3、-OH、-O-CO-(CH2)cCH3、-CdH2d-OH、-CdH2d-COOH、-CdH2d-2-COOH、-CdH2d-O-CO-(CH2)c-CH3，c＝8～20，d＝1～6。
R4Is selected from-H, -CeH2e-1or-CeH2e+1，e＝8～20。
In an embodiment of the present invention, it is,
m＝0～6；
R3selected from-OH, - (CH)2)2-COOH、-(CH2)4-O-CO-(CH2)14CH3or-O-CO- (CH)2)c-CH3；
R4Is selected from-CeH2e+1，e＝8～18；
In some embodiments of the present invention, the,
m＝0；
R3is- (CH)2)2-COOH；
R4is-CeH2e+1，e＝8～18。
In some embodiments of the present invention, the,
m＝1～6；
R3is-O-CO- (CH)2)c-CH3，c＝8～20；
R4is-CeH2e+1，e＝8～18。
The invention also provides an amphiphilic compound with a structure shown in the formula II, and the amphiphilic compound can be used for preparing a platinum complex:
R5is selected from-H, -CfH2f-1、-CfH2f+1、-Ar-CfH2f-1、-Ar-CfH2f+1、-Ar-O-CfH2f-1or-Ar-O-CfH2f+1，f＝1～22；
R6Is selected from-CgH2g-1、-CgH2g-2or-CgH2g-O-，g＝1～8；
R7Is selected from-OH and-ChH2h-1、-ChH2h+1、-Ar-ChH2h-1、-Ar-ChH2h+1、-Ar-O-ChH2h-1or-Ar-O-ChH2h+1，h＝1～22；
Ar is a monocyclic, bicyclic, and tricyclic carbocyclic ring system containing a total of 6-14 members, such as phenylene, naphthyl, anthracenylene, and the like, and at least one ring system is aromatic;
wherein any H attached to C may be substituted with a substituent;
wherein, the substituent is-OH, -NH2-COOH, halogen or-Ar;
R6and R7At least 1-COOH.
Or R6And R7The structure of (A) includes at least 1 carboxylate ion such as-COONa or-COOK.
In the amphiphilic compound provided by the invention,
R5is-CfH2f+1，R6Is composed ofR7is-OH;
or R5is-CfH2f+1，R6Is composed ofR7is-OH;
or R5is-CfH2f+1，R6Is composed ofR7is-ChH2h+1；
R5is-CfH2f+1，R6Is composed ofR7is-ChH2h+1；
f＝8～18；g＝2～6；h＝8～18。
The structural formula of the amphiphilic compound provided by the invention is shown as formulas II-a-II-b:
the amphiphilic compounds provided by the invention also comprise carboxylates of formulas II-a and II-b.
The carboxylic acid salts of the formula II-a and II-b included in the present invention include sodium stearyl fumarate, sodium stearyl maleate, sodium stearyl succinate, sodium lauryl maleate, potassium myristyl maleate, potassium palmityl maleate (but not limited to these salts), and also include organic base salts of these compounds.
The platinum complex provided by the invention is prepared by combining platinum compounds such as cisplatin, carboplatin, lobaplatin, oxaliplatin and nedaplatin (but not limited to the platinum drugs) with coordination bonds formed by carboxyl groups and carbonyl groups of ester bonds in an amphiphilic compound. The present invention has no special requirement on the reaction conditions, and any reaction conditions known in the art can be used for preparing the platinum complex.
The amphiphilic compound used in the platinum complex of formula I-a or formula I-b of the present invention can be prepared by esterification of one carboxyl group of dicarboxylic acid (such as fumaric acid, maleic acid, succinic acid, etc.) or anhydride (such as maleic anhydride, succinic anhydride, etc.) with fatty alcohol (but is not limited thereto, and all synthetic reactions that can generate the structure are possible). The amphiphilic compound in the platinum complexes shown in the formulas I-c and I-d is prepared by carrying out alcoholysis reaction on fatty acyl chloride and hydroxy acid (such as lactic acid, citric acid, salicylic acid, tartaric acid and the like, but not limited to the above compounds, compounds with similar structures can also be used). The reaction conditions of the present invention are not particularly limited, and any reaction conditions known in the art can be used for the preparation of esters by reacting an alcohol with an acid or an acid chloride.
The invention also provides a liposome of the platinum complex, which comprises the platinum complex and a lipid membrane material; the ratio of the platinum complex to the liposome membrane material is 99: 1-1: 99, and the preferable ratio is 49: 51-5: 95.
The ratio is a mass ratio, a molar ratio or a volume ratio.
In the liposome provided by the invention, the lipid membrane material comprises 1-100 parts by mass of phospholipid or/and 0-60 parts by mass of cholesterol.
In the liposome provided by the invention, the phospholipid includes one or more of lecithin, soybean phospholipid, hydrogenated soybean phospholipid, phosphatidylinositol, phosphatidylglycerol, dipalmitoylphosphatidylcholine, dimyristoylphosphatidylcholine, distearoylphosphatidylcholine, dilauroylphosphatidic acid, dilauroylphosphatidylglycerol, dimyristoylphosphatidylglycerol, palmitoylphosphatidylglycerol, distearoylphosphatidylglycerol, phosphatidylserine, dilauroylphosphatidylethanolamine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine, distearoylphosphatidylethanolamine and pegylated phospholipid.
The platinum complex provided by the invention can be added with proper auxiliary materials and additives, and prepared into different dosage forms by utilizing the known technology in the field, such as injection, freeze-dried powder injection for injection, liposome suspension, tablets, gel, implant and the like, and is used for intravenous injection, local injection, implantation, inhalation and other routes of administration.
The invention provides a medicament for treating cancer, which comprises the liposome provided by the invention.
The invention provides a compound with a structure shown in a formula II, which has good biocompatibility, carboxyl and carbonyl groups of ester bonds of the compound can be matched with platinum drugs, and unstable coordination bonds formed between the compound and the platinum drugs can increase the hydration speed of the drugs in vivo so as to improve the drug effect. Meanwhile, the platinum complex prepared from the compound provided by the invention can be well combined with a membrane material of the liposome, so that the entrapment rate and the drug-loading capacity of the liposome are improved. Moreover, after the platinum complex is prepared from the compound provided by the invention, the complex contains carboxyl, has pH sensitivity, and the carboxyl tends to be deprotonated in a lower pH value environment (such as tumor tissues), so that the release of a medicament in the tumor tissues is promoted, and the curative effect of the medicament is improved. The liposome provided by the invention can obviously reduce the toxic and side effects of the medicine and improve the medicine effect.
Drawings
FIG. 1-a shows a mass spectrum of the amphiphilic compound of example 1;
FIG. 1-b shows the mass spectrum of the amphiphilic compound of example 2;
FIG. 1-c shows the mass spectrum of the amphiphilic compound of example 3;
FIG. 1-d shows the mass spectrum of the amphiphilic compound of example 4;
FIG. 1-e shows a mass spectrum of the amphiphilic compound of example 5;
FIG. 2-a shows the nuclear magnetic spectrum of the amphiphilic compound of example 1;
FIG. 2-b shows the nuclear magnetic spectrum of the amphiphilic compound of example 2;
FIG. 2-c shows the nuclear magnetic spectrum of the amphiphilic compound of example 3;
FIG. 2-d shows the nuclear magnetic spectrum of the amphiphilic compound of example 4;
FIG. 3-a shows an infrared spectrum of the amphiphilic compound of example 1;
FIG. 3-b shows an infrared spectrum of the amphiphilic compound of example 2;
FIG. 3-c shows an infrared spectrum of the amphiphilic compound of example 4;
FIG. 3-d shows an infrared spectrum of the amphiphilic compound of example 5;
figure 4-a shows the XRD pattern of the amphiphilic compound of example 1;
figure 4-b shows the XRD pattern of the amphiphilic compound of example 2;
figure 4-c shows the XRD pattern of the amphiphilic compound of example 3;
figure 4-d shows the XRD pattern of the amphiphilic compound of example 4;
figure 4-e shows the XRD pattern of the amphiphilic compound of example 5;
FIG. 5 shows an electron microscopy of liposomes prepared according to example 24;
FIG. 6 is a graph showing the particle size distribution of liposomes obtained in example 24;
FIG. 7 shows tumor-bearing mouse survival rates (7a-7 d);
FIG. 8 shows tumor-bearing mouse kidney tissue sections;
FIG. 9 shows survival of mice in preliminary safety assessments;
FIG. 10 shows the rate of change of body weight of mice in the preliminary safety evaluation.
Detailed Description
The invention provides a compound, application thereof, a platinum complex and a liposome thereof, and a person skilled in the art can realize the compound by appropriately improving process parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to those skilled in the art are deemed to be included within the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The invention provides a platinum complex with a structure shown in formulas I-a-I-d:
R1is selected from-CnH2n-、-CnH2n-2-、-Ar-CnH2n-、Ar-CnH2n-2-、-Ar-O-CnH2n-、Ar-O-CnH2n-2-，n＝0～22；
R2Is selected from-H, -CbH2b-1、-CbH2b+1、-Ar-CbH2b-1、-Ar-CbH2b+1、-Ar-O-CbH2b-1or-Ar-O-CbH2b+1，b＝1～22；
R3Is selected from-NH2、-(CH2)c-NHCO-(CH2)dCH3、-OH、-O-CO-(CH2)cCH3、-CdH2d-OH、-CdH2d-2-OH、-CdH2d-COOH、-CdH2d-2-COOH、-CdH2d-O-CO-(CH2)c-CH3、，c＝1～21，d＝1～8；
R4Is selected from-H, -CeH2e-1、-CeH2e+1、-Ar-CeH2e-1、-Ar-CeH2e+1、-Ar-O-CeH2e-1or-Ar-O-CeH2e+1，e＝1～22；
m＝0～10；
Wherein any H attached to C may be substituted with a substituent;
wherein, the substituent is-OH, -NH2-COOH, halogen or-Ar.
In the present invention, -CbH2b+1or-CeH2e+1Is alkyl, and the value of b is 1-22. In some embodiments, b is 8-20. In other embodiments, b is 8-18 or b is 12-18; in other embodiments, b is 14-16; in some embodiments, b is 15. The value of e and the value of b can be the same or different and are independent, and the value of e is 1-22. In some embodiments, e is 8-20. In other embodiments, e is 8-18 or e is 12-18; in other embodiments, e is 14-16; in some embodiments, e is 15. The alkyl is a straight-chain alkyl or branched-chain alkyl; in some embodiments, it is a straight chain alkyl group. The alkyl group is a substituted or unsubstituted alkyl group. The substituent means that H connected to any C in the alkyl can be substituted by a substituent, and the substituent of the substituted alkyl is-NH2One or more of-OH, -COOH, halogen or-Ar. The number of the substituents on any C is 1 or two. In the present invention, -CbH2b+1Alkyl groups include, but are not limited to, methyl (Me, -CH)3) Ethyl (Et, -CH)2CH3) N-propyl (n-Pr, -CH)2CH2CH3) Isopropyl (i-Pr, -CH (CH)3)2) N-butyl (n-Bu, -CH)2CH2CH2CH3) Isobutyl (i-Bu, -CH)2CH(CH3)2) Sec-butyl (s-Bu, -CH (CH)3)CH2CH3) Tert-butyl (t-Bu, -C (CH)3)3) N-pentyl (-CH)2CH2CH2CH2CH3) 2-pentyl (-CH (CH)3)CH2CH2CH3) 3-pentyl (-CH (CH)2CH3)2) 2-methylbutyl (-CH)2CH(CH3)CH2CH3) 2-methyl-2-butyl (-C (CH)3)2CH2CH3) 3-methylbutyl (-CH)2CH2CH(CH3)2) 3-methyl-2-butyl (-CH (CH)3)CH(CH3)2) N-hexyl (-CH)2CH2CH2CH2CH2CH3) 2-hexyl (-CH (CH)3)CH2CH2CH2CH3) 3-hexyl (-CH (CH)2CH3)CH2CH2CH3) 2-methylpentyl (-CH)2CH(CH3)CH2CH2CH3) 2-methyl-2 pentyl (-CH (CH)3)2CH2CH2CH3) 2-methyl-3 pentyl (-CH (CH)2CH3)CH(CH3)2) 3-methylpentyl (-CH)2CH2CH(CH3)CH2CH3) 3-methyl-2-pentyl (-CH (CH)3)CH(CH3)CH2CH3) 3-methyl-3-pentyl (-CH (CH)3)(CH2CH3)2) 4-methyl-2 pentyl (-CH (CH)3)CH2CH(CH3)2) 4-first of allCyclopentyl (-CH)2CH2CH2CH(CH3)2) 2, 2-dimethylbutyl (-CH)2C(CH3)2CH2CH3) 3, 3-dimethyl-2-butyl (-CH (CH)3)C(CH3)3) 3, 3-dimethylbutyl (-CH)2CH2C(CH3)3) 2, 3-dimethylbutyl (-CH)2CH(CH3)CH(CH3)2) 2, 3-dimethyl-2-butyl (-C (CH)3)2CH(CH3)2) N-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecyl, n-hexadecyl, n-heptadecyl, n-octadecyl.
In the present invention, -CbH2b-1or-CeH2e-1Refers to alkenyl groups in which at least one position is unsaturated, i.e. one C-C is sp2A double bond. The value of b is 2-22. In some embodiments, b is 8-20. In other embodiments, b is 8-18 or b is 12-18; in other embodiments, b is 14-16; in some embodiments, b is 15. The value of e and the value of b can be the same or different and are independent, and the value of e is 2-22. In some embodiments, e is 8-20. In other embodiments, e is 8-18 or e is 12-18; in other embodiments, e is 14-16; in some embodiments, e 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. Any H connected to C in the substituted alkenyl can be substituted by a substituent, and the substituent of the substituted alkenyl is-NH2One or more of-OH, -COOH, halogen or-Ar. The number of the substituents on any C is 1 or two. The alkenyl group includes the orientation of the groups "trans", "n" or "E" and "Z", wherein specific examples include, but are not limited to, vinyl (-CH. CH)2) Allyl, allyl (-CH)2CH＝CH2) Butenyl and 4-methylbutenyl, 2-pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenylAlkenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl.
In the present invention, -Ar-CbH2b+1Means a group-C attached to a phenyl or aryl groupbH2b+1，-Ar-CbH2b-1Means a group-C attached to a phenyl or aryl groupbH2b-1，-Ar-O-CbH2b+1Refers to-C attached to-Ar-O-bH2b+1，-Ar-O-CbH2b-1Refers to-C attached to-Ar-O-bH2b-1。-Ar-CeH2e+1Means a group-C attached to a phenyl or aryl groupeH2e+1，-Ar-CeH2e-1Means a group-C attached to a phenyl or aryl groupeH2e-1，-Ar-O-CeH2e+1Refers to-C attached to-Ar-O-eH2e+1，-Ar-O-CeH2e-1Refers to-C attached to-Ar-O-eH2e-1. the-Ar-represents monocyclic, bicyclic, and tricyclic carbon ring systems containing a total of 6-14 membered rings, and at least one ring system is aromatic, wherein each ring system comprises 3-7 membered rings, wherein there are two points of attachment to the rest of the molecule. Ar-can be phenylene, naphthyl or anthrylene. And said-Ar-may be substituted or unsubstituted.
In the present invention, -CnH2n-or-CdH2d-The value of n is 1-8. In some embodiments, n is between 1 and 6. In other embodiments, n is 1-4; in other embodiments, n is 2-4; the alkylene group is a linear or branched alkylene group; the value of d can be the same as or different from that of n and is independent, and the value of d is 1-8. In some embodiments, d is between 1 and 6. In other embodiments, d is 1-4; in other embodiments, d is 2 to 4; the alkylene group is a linear or branched alkylene group; in some embodiments, it is a linear alkylene group. The alkylene group is a substituted or unsubstituted alkylene group. The substituent means that H connected with any C in the alkylene can be substituted by substituent, and the substituent of the substituted alkyl is-NH2One or more of-OH, -COOH or-Ar. The number of the substituents on any C is 1 or two. In the present invention, alkylene includes, but is not limited to, methylene (-CH)2-, ethylene (-CH)2CH2-, n-propylene (-CH)2CH2CH2-, isopropylidene (-C (CH)3)2-, n-butylidene (-CH)2CH2CH2CH2-), isobutylene, sec-butylene, tert-butylene, n-pentylene, 2-pentylene, 3-pentylene, 2-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, 3-dimethyl-2-butylene, 3, 3-dimethyl-2-butylene, n-heptylene, n-octylene, carboxyl-substituted propylene, carboxyl-substituted n-butylene, carboxyl-substituted n-pentylene, amino-substituted propylene, amino-substituted n-butylene, amino-substituted n-pentylene.
In the present invention, -CnH2n-2-or-CdH2d-2-denotes alkenylene in which at least one position is unsaturated, i.e. one C-C is sp2A double bond. The value of n is 2-8. In some embodiments, n is 2-6. In other embodiments, n is 2-4. The value of d can be the same as or different from that of n and is independent, and the value of d is 2-8. In some embodiments, d is between 2 and 6. In other embodiments, d is 2-4. The alkenyl group is a linear alkenylene group or a branched alkenylene group, and in some embodiments, is a linear alkenylene group. The alkenylene group is a substituted or unsubstituted alkenylene group. Any H connected to C in the substituted alkenylene can be substituted by a substituent, and the substituent of the substituted alkenylene is-NH2One or more of-OH, -COOH or-Ar. The number of the substituents on any C is 1 or two. The alkenylene group includes the positioning of the groups "trans", "n" or "E" and "Z", and specific examples thereof include, but are not limited to, ethenylene (-CH-), propenylene (-CH-), allylene (-CH-)2CH-CH-), butenylene and 4-methylbutenylene, 2-pentenylene, hexenylene, heptenylene, octenylene.
In some embodiments of the present invention, the,
R1is selected from-CnH2n-or-CnH2n-2，n＝1～10；
R2Is selected from-CbH2b-1or-CbH2b+1，b＝8～20；
R3Selected from-OH, -O-CO- (CH)2)cCH3、-CdH2d-OH、-CdH2d-COOH、-CdH2d-2-COOH、-CdH2d-O-CO-(CH2)c-CH3、，c＝8～20，d＝1～6。
R4Is selected from-H, -CeH2e-1or-CeH2e+1，e＝8～20。
In some embodiments of the present invention, the,
m＝0～6；
R1is selected from-CH2-、
R2Is selected from-CbH2b+1，b＝12～18；
R3Is- (CH)2)2-COOH、-CH2-COOH、-(CH2)c-CO-(CH2)c-CH3，c＝8～18；
R4is-CeH2e-1or-CeH2e+1，e＝12～18。
In some embodiments, R1Is composed ofR2Is- (CH)2)17CH3。
In some embodiments, m is 0; r3Is- (CH)2)2-COOH；R4is-CeH2e+1，e＝8～18。
The platinum complex provided by the invention is represented by the formula (1) to (18), wherein x is 7-17:
in the structural formulas (1) to (18) of the platinum complex provided by the invention, x is 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17.
The platinum complex provided by the invention is prepared by reacting a compound shown in a formula II or a compound shown in a formula III with a platinum compound; the platinum compound is cisplatin, carboplatin, oxaliplatin or nedaplatin, eptaplatin, lobaplatin, etc.
R5Is selected from-H, -CfH2f+1、-CfH2f-1、-Ar-CfH2f+1、-Ar-CfH2f-1、-Ar-O-CfH2f+1or-Ar-OCfH2f-1,f＝1～22；
R6Is selected from-CgH2g-、-CgH2g-2-、-CgH2g-O-or-CgH2g-2-O-,g＝1～8；
R7Is selected from-OH and-ChH2h+1、-ChH2h-1、-Ar-ChH2h+1、-Ar-ChH2h-1、-Ar-O-ChH2h+1or-Ar-OChH2h-1,h＝1～22；
R8Is selected from-CiH2i-、-CiH2i-2-、-Ar-CiH2i-、-Ar-CiH2i-2-、-Ar-O-CiH2i-、-Ar-O-CiH2i-2-,i＝0～22；
R9Is selected from-H, -CpH2p+1、-CpH2p-1、-Ar-CpH2p+1、-Ar-CpH2p-1、-Ar-O-CpH2p+1、-Ar-O-CpH2p-1,p＝0～22；
Wherein any H attached to C may be substituted with a substituent;
wherein, the substituent is-OH, -NH2-COOH, halogen or-Ar.
R6And R7At least 1-COOH.
In the method provided by the invention, the molar ratio of the compound shown in the formula II or the formula III to Pt in the platinum compound is not more than 30: 1.
In some embodiments, the molar ratio of the compound of formula II or III to Pt in the platinum-based compound is (1-8): 1.
in other embodiments, the molar ratio of the compound of formula II or III to Pt in the platinum-based compound is (1-5): 1.
in other embodiments, the molar ratio of the compound of formula II or formula III to Pt in the platinum-based compound is 2: 1.
in other embodiments, the molar ratio of the compound of formula II or formula III to Pt in the platinum-based compound is 1: 1.
the solvent adopted in the reaction for preparing the platinum complex is an aqueous medium.
In the examples provided by the present invention, the aqueous medium is water, an aqueous mannitol solution, an aqueous sodium chloride solution, an aqueous glycerol solution, an aqueous sugar solution, a phosphate buffer solution, or the like (without limitation), and the aqueous medium may be the above-mentioned aqueous solution containing Dimethylformamide (DMF), Dimethylsulfoxide (DMSO), or ethanol.
The invention provides an amphiphilic compound having a structure represented by formula II:
R5is selected from-H, -CfH2f+1、-CfH2f-1、-Ar-CfH2f+1、-Ar-CfH2f-1、-Ar-O-CfH2f+1or-Ar-OCfH2f-1,f＝1～22；
R6Is selected from-CgH2g-、-CgH2g-2-、-CgH2g-O-or-CgH 2g-2-O-, g ═ 1 to 8;
R7is selected from-OH and-ChH2h+1、-ChH2h-1、-Ar-ChH2h+1、-Ar-ChH2h-1、-Ar-O-ChH2h+1or-Ar-OChH2h-1,h＝1～22；
Wherein any H attached to C may be substituted with a substituent;
wherein, the substituent is-OH, -NH2-COOH, halogen or-Ar;
R6and R7At least 1-COOH.
In the present invention, -CfH2f+1or-ChH2h+1Is an alkyl group, and f is 1-22. In some embodiments, f is 8-20. In other embodiments, f is 8-18 or f is 12-18; in other embodiments, f is 14-16; in some embodiments, f is 15. The value of h and the value of f can be the same or different and are independent, and the value of h is 1-22. In some embodiments, h is 8-20. In other embodiments, h is 8-18 or h is 12-18; in other embodiments, h is 14-16; in some embodiments, h is 15. The alkyl is a straight-chain alkyl or branched-chain alkyl; in some embodiments, it is a straight chain alkyl group. The alkyl group is a substituted or unsubstituted alkyl group. The substituent means that H connected to any C in the alkyl group may be substituted by a substituentAnd the substituent of the substituted alkyl is-NH2One or more of-OH, -COOH, halogen or-Ar. The number of the substituents on any C is 1 or two. In the present invention, -CbH2b+1Alkyl groups include, but are not limited to, methyl (Me, -CH)3) Ethyl (Et, -CH)2CH3) N-propyl (n-Pr, -CH)2CH2CH3) Isopropyl (i-Pr, -CH (CH)3)2) N-butyl (n-Bu, -CH)2CH2CH2CH3) Isobutyl (i-Bu, -CH)2CH(CH3)2) Sec-butyl (s-Bu, -CH (CH)3)CH2CH3) Tert-butyl (t-Bu, -C (CH)3)3) N-pentyl (-CH)2CH2CH2CH2CH3) 2-pentyl (-CH (CH)3)CH2CH2CH3) 3-pentyl (-CH (CH)2CH3)2) 2-methylbutyl (-CH)2CH(CH3)CH2CH3) 2-methyl-2-butyl (-C (CH)3)2CH2CH3) 3-methylbutyl (-CH)2CH2CH(CH3)2) 3-methyl-2-butyl (-CH (CH)3)CH(CH3)2) N-hexyl (-CH)2CH2CH2CH2CH2CH3) 2-hexyl (-CH (CH)3)CH2CH2CH2CH3) 3-hexyl (-CH (CH)2CH3)CH2CH2CH3) 2-methylpentyl (-CH)2CH(CH3)CH2CH2CH3) 2-methyl-2 pentyl (-CH (CH)3)2CH2CH2CH3) 2-methyl-3 pentyl (-CH (CH)2CH3)CH(CH3)2) 3-methylpentyl (-CH)2CH2CH(CH3)CH2CH3) 3-methyl-2-pentyl (-CH (CH)3)CH(CH3)CH2CH3) 3-methyl-3-pentyl (-CH (CH)3)(CH2CH3)2) 4-methyl-2 pentyl (-CH (CH)3)CH2CH(CH3)2) 4-methylpentyl (-CH)2CH2CH2CH(CH3)2) 2, 2-dimethylbutyl (-CH)2C(CH3)2CH2CH3) 3, 3-dimethyl-2-butyl (-CH (CH)3)C(CH3)3) 3, 3-dimethylbutyl (-CH)2CH2C(CH3)3) 2, 3-dimethylbutyl (-CH)2CH(CH3)CH(CH3)2) 2, 3-dimethyl-2-butyl (-C (CH)3)2CH(CH3)2) N-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecyl, n-hexadecyl, n-heptadecyl, n-octadecyl.
In the present invention, -CfH2f-1or-ChH2h-1Refers to alkenyl groups in which at least one position is unsaturated, i.e. one C-C is sp2A double bond. The value of f is 2-22. In some embodiments, f is 8-20. In other embodiments, f is 8-18 or f is 12-18; in other embodiments, f is 14-16; in some embodiments, f is 15. The value of h and the value of f can be the same or different and are independent, and the value of h is 2-22. In some embodiments, h is 8-20. In other embodiments, h is 8-18 or h is 12-18; in other embodiments, h is 14-16; in some embodiments, h 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 connected with any C in the substituted alkenyl can be substituted by substituent, and the substituent of the substituted alkenyl is one or more of-NH 2, -OH, -COOH, halogen or-Ar. The number of the substituents on any C is 1 or two. The alkenyl group includes the orientation of the groups "trans", "n" or "E" and "Z", wherein specific examples include, but are not limited to, vinyl (-CH. CH)2) Allyl, allyl (-CH)2CH＝CH2) Butenyl and 4-methylbutenyl, 2-pentenyl, hexenyl, heptenyl, octenylAlkenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl.
In the present invention, -Ar-CfH2f+1Means a group-C attached to a phenyl or aryl groupfH2f+1，-Ar-CfH2f-1Means a group-C attached to a phenyl or aryl groupfH2f-1，-Ar-O-CfH2f+1Refers to-C attached to-Ar-O-fH2f+1，-Ar-O-CfH2f-1Refers to-C attached to-Ar-O-fH2f-1。-Ar-ChH2h+1Means a group-C attached to a phenyl or aryl grouphH2h+1，-Ar-ChH2h-1Means a group-C attached to a phenyl or aryl grouphH2h-1，-Ar-O-ChH2h+1Refers to-C attached to-Ar-O-hH2h+1，-Ar-O-ChH2h-1Refers to-C attached to-Ar-O-hH2h-1. the-Ar-represents monocyclic, bicyclic, and tricyclic carbon ring systems containing a total of 6-14 membered rings, and at least one ring system is aromatic, wherein each ring system comprises 3-7 membered rings, wherein there are two points of attachment to the rest of the molecule. Ar-can be phenylene, naphthyl or anthrylene. And said-Ar-may be substituted or unsubstituted.
In the present invention, -CgH2g-is an alkylene group, g is 1-8. In some embodiments, g is between 1 and 6. In other embodiments, g is 1-4; in other embodiments, g is 2-4; the alkylene group is a linear or branched alkylene group; in some embodiments, it is a linear alkylene group. The alkylene group is a substituted or unsubstituted alkylene group. The substituent means that H connected with any C in the alkylene can be substituted by substituent, and the substituent of the substituted alkyl is-NH2One or more of-OH, -COOH or-Ar. The number of the substituents on any C is 1 or two. In the present invention, alkylene includes, but is not limited to, methylene (-CH)2-, ethylene (-CH)2CH2-, n-propylene (-CH)2CH2CH2-, isopropylidene (-C (CH)3)2-, n-butylidene (-CH)2CH2CH2CH2-), isobutylene, sec-butylene, tert-butylene, n-pentylene, 2-pentylene, 3-pentylene, 2-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, 3-dimethyl-2-butylene, 3, 3-dimethyl-2-butylene, n-heptylene, n-octylene, carboxyl-substituted propylene, carboxyl-substituted n-butylene, carboxyl-substituted n-pentylene, amino-substituted propylene, amino-substituted n-butylene, amino-substituted n-pentylene.
In the present invention, -CgH2g-2-denotes alkenylene in which at least one position is unsaturated, i.e. one C-C is sp2A double bond. The value of g is 2-8. In some embodiments, g is between 2 and 6. In other embodiments, g is 2-4. The alkenyl group is a linear alkenylene group or a branched alkenylene group, and in some embodiments, is a linear alkenylene group. The alkenylene group is a substituted or unsubstituted alkenylene group. Any H connected to C in the substituted alkenylene can be substituted by a substituent, and the substituent of the substituted alkenylene is-NH2One or more of-OH, -COOH or-Ar. The number of the substituents on any C is 1 or two. The alkenylene group includes the positioning of the groups "trans", "n" or "E", "Z", and specific examples thereof include, but are not limited to, ethenylene (-CH-), propenylene (-CH-), allylene (-CH-)2CH ═ CH-), butenylene and 4-methylbutenylene, 2-pentenylene, hexenylene, heptenylene, octenylene.
In the present invention, -CgH2g-O-or-CgH2g-2-O-means a group-C linked to-O-)gH2g-or-CgH2g-2-。
In an embodiment of the invention, in the amphiphilic compound:
R1is selected from-CfH2f+1、-CfH2f-1，f＝8～18；
R2Is selected from-CgH2g-、-CgH2g-1-NH-，g＝2～6；
R3Is selected from-OH and-ChH2h+1、-ChH2h-1，h＝8～18。
Wherein any H attached to C may be substituted with one or two substituents;
wherein the substituent is-NH2or-COOH.
The R is2And R3At least 1-COOH.
In an embodiment of the present invention, it is,
R5is-CfH2f+1，R6Is composed ofR7is-OH;
or R5is-CfH2f+1，R6Is composed ofR7is-OH;
or R5is-CfH2f+1，R6Is composed ofR7is-ChH2h+1；
Or R5is-CfH2f+1，R6Is composed ofR7is-ChH2h+1；
f＝8～18；g＝2～6；h＝8～18。
In the embodiment of the invention, the structure is as follows:
the platinum complex prepared by the compound provided by the invention can be well combined with a membrane material of a liposome, so that the entrapment rate and the drug-loading capacity of the liposome are improved. Moreover, after the compound provided by the invention is used for preparing the platinum complex, the complex contains carboxyl, has pH sensitivity, and the carboxyl tends to be deprotonated in a lower pH value environment (such as tumor tissues), so that the release of the medicament in the tumor tissues is promoted, the curative effect of the medicament is improved, and the toxic or side effect of the medicament is reduced.
The invention also provides a liposome of the platinum complex, which comprises the platinum complex and a lipid membrane material; the ratio of the platinum complex to the liposome membrane material is 99: 1-1: 99.
The ratio is a mass ratio, a molar ratio or a volume ratio.
In the liposome provided by the invention, the lipid membrane material comprises 1-100 parts by mass of phospholipid or/and 0-60 parts by mass of cholesterol.
In the liposome provided by the invention, the phospholipid includes one or more of lecithin, soybean phospholipid, hydrogenated soybean phospholipid, phosphatidylinositol, phosphatidylglycerol, dipalmitoylphosphatidylcholine, dimyristoylphosphatidylcholine, distearoylphosphatidylcholine, dilauroylphosphatidic acid, dilauroylphosphatidylglycerol, dimyristoylphosphatidylglycerol, palmitoylphosphatidylglycerol, distearoylphosphatidylglycerol, phosphatidylserine, dilauroylphosphatidylethanolamine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine, distearoylphosphatidylethanolamine and pegylated phospholipid.
The liposome provided by the invention optionally also comprises long-circulating material, and the long-circulating material adopted by the invention is MPEG2000-DSPE，MPEG5000-DSPE、MPEG2000-DMPE，MPEG5000-DMPE，MPEG2000-DPPE，MPEG5000DPPE and the like.
The mass fraction of the long-circulating material in the liposome is 0-50%.
Furthermore, the mass fraction of the long-circulating material in the liposome provided by the invention is 6-35%.
The liposome provided by the invention can also comprise an antioxidant; the antioxidant adopted by the invention is any one or a composition of more than two of L-cysteine, ascorbic acid, dl-alpha-tocopherol, sodium sulfite, sodium metabisulfite or anhydrous sodium bisulfite.
In the liposome provided by the invention, the mass fraction of the antioxidant is not higher than 20%.
The liposome provided by the invention can also be added with an isoosmotic adjusting agent, wherein the isoosmotic adjusting agent is a composition of one or more than two of mannitol, sorbitol, glucose, sucrose or sodium chloride.
In some embodiments, the liposome is prepared by: quickly injecting a proper amount of solution containing a membrane material, an antioxidant or/and a long-circulating material and containing an organic solvent into the aqueous solution of the platinum complex, stirring for 30min at 40-50 ℃, and homogenizing by a liposome extruder to obtain the novel platinum liposome. In addition, the long circulating material may be added after liposome formation to modify the liposome surface.
In other embodiments, the liposomes are prepared by: adding a proper amount of organic solvent to dissolve a membrane material, an antioxidant and a long-circulating material in a round-bottom flask, adding an aqueous solution of the platinum complex provided by the invention, homogenizing mechanically or ultrasonically to form a W/O emulsion, removing the organic solvent by reduced pressure rotary evaporation, forming a thin film on the wall of the container, injecting a hydration medium, hydrating the membrane material for 1-2 h at 40-55 ℃, and homogenizing by a liposome extruder to obtain the novel platinum liposome.
In other embodiments, the liposomes are prepared by: adding a proper amount of organic solvent into a round-bottom flask for dissolving, performing reduced pressure rotary evaporation to remove the organic solvent, forming a thin film on the wall of the container, injecting a hydration medium to hydrate the film for 1-2 h, and homogenizing by a liposome extruder or microjet to obtain the novel platinum liposome.
In other embodiments, the liposomes are prepared by: dissolving a membrane material, an antioxidant and a long-circulating material in a round-bottom flask by adding a proper amount of organic solvent, adding the platinum compound provided by the invention, ultrasonically or mechanically homogenizing to form a W/O emulsion, removing part of the organic solvent by reduced pressure rotary evaporation, injecting a hydration medium, mechanically stirring to form W/O/W, removing the organic solvent by reduced pressure rotary evaporation, and homogenizing and dispersing by a liposome extruder and microjet to obtain the novel platinum liposome.
The invention provides a medicament for treating cancer, which comprises the liposome provided by the invention.
The medicine provided by the invention can be used for treating cancers, 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 dosage form of the medicine provided by the invention is injection (suspension type or freeze-dried powder for injection), emulsion, submicron emulsion and the like.
Experiments show that the liposome provided by the invention can obviously improve the blood concentration of the platinum compound, prolong the residence time of the medicament in vivo, and concentrate the platinum medicament in a cancerous organ, so that the dosage of the medicament can be reduced, the curative effect can be improved, the toxic and side effects of the medicament can be reduced, and the liposome is suitable for treating various tumor diseases.
The invention provides a compound with a structure shown in a formula II, which has good biocompatibility, carboxyl and carbonyl groups of ester bonds of the compound can be matched with platinum drugs, and unstable coordination bonds formed between the compound and the platinum drugs can increase the hydration speed of the drugs in vivo so as to improve the drug effect. Meanwhile, the platinum complex prepared from the compound provided by the invention can be well combined with a membrane material of the liposome, so that the entrapment rate and the drug-loading capacity of the liposome are improved. Moreover, after the platinum complex is prepared from the compound provided by the invention, the compound contains carboxyl, has pH sensitivity, and the carboxyl tends to be deprotonated in a low pH value environment (such as tumor tissues), so that the compound is favorable for promoting the release of a medicament in the tumor tissues. The liposome provided by the invention can reduce the toxic and side effects of the medicine, especially the renal toxicity, and keep a certain curative effect.
The reagents or instruments adopted by the invention are all common products sold on the market and can be purchased on the market.
Example 1 amphiphilic compound and preparation thereof:
the preparation method comprises the following steps:
1. examples 1 to 2
Adding 1.4mol of maleic anhydride, 1.0mol of octadecanol and 1000mL of cyclohexane into a reaction vessel, dropwise adding 13.50g of triethylamine at room temperature, heating to 60 ℃, reacting for 4 hours, and distilling under reduced pressure until the mixture is dry; then adding 900mL of acetone into the residue, heating to 56 ℃, stirring for 30min, cooling to 40 ℃, stirring for 1h, then distilling under reduced pressure to obtain an oily liquid, cooling to room temperature, adding a proper amount of water to precipitate a solid, stirring for 1h, performing suction filtration, washing with water and petroleum ether for several times respectively, and performing vacuum drying on the obtained solid for 20h at 60 ℃, thus obtaining the amphiphilic compound in the embodiment 1-2. The obtained amphiphilic compound was subjected to mass spectrometry, and the results are shown in FIGS. 1-a to 1-b.
Or adding 1.2mol of anhydride (maleic anhydride or succinic anhydride), 1.0mol of aliphatic alcohol (hexadecanol or octadecanol) and 1000mL of cyclohexane into a reaction vessel, dropwise adding 8.1g of triethylamine at room temperature, heating to 50 ℃ for reaction for 4 hours, and then distilling under reduced pressure to remove the cyclohexane to obtain a solid; and then adding 800mL of absolute ethyl alcohol into the obtained solid, heating to 60 ℃ for dissolution, stirring for 30min, cooling to 40 ℃, stirring for 1h, then distilling under reduced pressure to obtain oily liquid, cooling to room temperature, adding a proper amount of water to separate out the solid, stirring for 1h, performing suction filtration, washing with water and petroleum ether for several times respectively, and performing vacuum drying on the obtained solid for 20h at 60 ℃ to obtain the amphiphilic compound in the embodiment 1-2. The obtained amphiphilic compound was characterized by nuclear magnetism, infrared and XRD, and the results are shown in FIGS. 2-a-2-b, 3-a-3-b and 4-a-4-b.
2. Examples 3 to 6
Adding 1.4mol of anhydride (maleic anhydride or succinic anhydride), 1.0mol of aliphatic alcohol (hexadecanol or octadecanol) and 1000mL of cyclohexane into a reaction vessel, dropwise adding 13.50g of triethylamine at room temperature, heating to 60 ℃, reacting for 4 hours, and distilling under reduced pressure until the mixture is dry; and then adding 900mL of acetone into the residue, heating to 56 ℃, stirring for 30min, cooling to 40 ℃, stirring for 1h, then adding a proper amount of water, cooling to room temperature, stirring for 1h, separating out a solid, performing suction filtration, washing with acetone, and performing vacuum drying on the obtained solid for 20h at 60 ℃ to obtain the amphiphilic compound in the embodiment 3-6. Mass spectrum characterization is performed on the prepared amphiphilic compounds of examples 3-5, the results are shown in figures 1-c-1-e, nuclear magnetic characterization is performed on examples 3-4, the results are shown in figures 2-c-2-d, and infrared spectrum characterization is performed on examples 4-5, the results are shown in figures 3-c-3-d.
Or adding 1.2mol of anhydride (maleic anhydride or succinic anhydride), 1.0mol of aliphatic alcohol (hexadecanol or octadecanol) and 1000mL of cyclohexane into a reaction vessel, dropwise adding 8.1g of triethylamine at room temperature, heating to 50 ℃ for reaction for 4 hours, and then distilling under reduced pressure to remove the cyclohexane to obtain a solid; and then adding 800mL of absolute ethyl alcohol into the obtained solid, heating to 60 ℃ for dissolution, stirring for 30min, cooling to 40 ℃, preserving heat for 1h, then adding a proper amount of water until a small amount of solid is separated out, stirring for 1h, cooling to room temperature, separating out the solid, performing suction filtration, washing with ethanol, and performing vacuum drying on the obtained solid for 20h at 60 ℃ to obtain the amphiphilic compound in the embodiment 3-5. XRD characterization is carried out on the prepared amphiphilic compound in the embodiment 3-5, and the result is shown in figures 4-c-4-e.
Examples 7 to 20 preparation of platinum Complex
TABLE 1 examples 7 to 20
The preparation method comprises the following steps: adding an amphiphilic compound into water, stirring and dissolving under a certain condition, adding a platinum compound (the molar ratio of the amphiphilic compound to the platinum compound is 1:1-10:1), heating and stirring for 4 hours, reacting to obtain a platinum complex, and drying to obtain solid powder of the platinum complex.
Examples 21-31 reverse Evaporation method for preparing liposomes
TABLE 2 examples 21 to 31
Placing the film material and the antioxidant in a formula amount into a round-bottom flask, adding a proper amount of organic solvent (the organic solvent is chloroform or water saturated ether or a mixed solvent of at least two solvents of dichloromethane, methanol and ethanol), dissolving, adding a formula amount of platinum complex aqueous solution (the volume ratio of the organic solvent to the water phase is 3: 1-6: 1), homogenizing mechanically or ultrasonically to form W/O emulsion, removing the organic solvent by reduced pressure rotary evaporation, forming a film on the wall of the container, injecting hydration medium (normal saline, 5% glucose, 5% sucrose, 7% sucrose, water for injection or 3% mannitol aqueous solution), hydrating the film material for 1-2 h at 40 (or 55 ℃), homogenizing by a liposome extruder or a microjet to obtain platinum complex liposome, adding long-circulating material, stirring at room temperature for 1h, and obtaining the platinum complex liposome.
Examples 32 to 34 preparation of liposomes by ethanol injection
TABLE 3 examples 32 to 34
Putting the platinum complex with the formula amount into a round-bottom flask, adding a proper amount of water and 7% of sucrose, dissolving, quickly injecting an ethanol solution of the membrane material with the formula amount, stirring for 30min at 40-50 ℃, homogenizing by a liposome extruder, and adding a proper amount of water and 7% of sucrose to dilute to a certain concentration to obtain the novel platinum liposome.
Example 35 preparation of liposomes by film Dispersion
Table 4 example 35
Placing the film material, the antioxidant, the long-circulating material and the platinum complex in the formula amount into a round-bottom flask, adding a proper amount of chloroform for dissolving, performing reduced pressure rotary evaporation to remove the organic solvent, forming a thin film on the wall of the container, injecting a hydration medium (water for injection), hydrating the film material for 1-2 hours at 40-55 ℃, and homogenizing by a liposome extruder to obtain the novel platinum liposome.
EXAMPLE 36 multiple emulsion method for preparing liposomes
TABLE 5 example 36
Placing the membrane material and the antioxidant in a formula amount into a round-bottom flask, adding a proper amount of diethyl ether, dissolving, adding a platinum compound aqueous solution in a formula amount (the volume ratio of an organic phase to a water phase is 3:1), ultrasonically or mechanically homogenizing to form a W/O emulsion, performing reduced pressure rotary evaporation to remove part of the organic solvent, injecting a 7% sucrose aqueous solution, mechanically stirring to form W/O/W, performing reduced pressure rotary evaporation to remove the organic solvent, and homogenizing and dispersing by a liposome extruder and a microjet to obtain the novel platinum liposome.
Example 37 Liposome quality examination
The morphology of the platinum complex-containing liposome prepared by the invention is observed by an electron microscope, and the particle size and the distribution of the liposome are measured by a Zetasizer NanoZS90 laser particle size analyzer. The encapsulation efficiency of the liposome prepared by the invention is calculated by a method provided in Chinese pharmacopoeia. The results are shown in Table 6:
TABLE 6 examination of liposome particle size and encapsulation efficiency
Example number
Average particle diameter (nm)
Encapsulation efficiency (%)
21
123.4
91.3
22
119.6
95.1
23
102.8
94.5
24
107.7
94.4
25
106.5
93.6
26
117.3
91.2
27
102.2
98.1
28
96.7
92.5
29
109.4
97.2
30
115.8
92.6
31
114.1
94.9
32
96.4
90.1
33
105.3
92.5
34
124.8
90.6
FIGS. 5 and 6 show the morphology and size distribution of the liposomes obtained in example 24, which were examined similarly to the liposomes obtained in other examples.
The result shows that the particle size of the novel platinum complex liposome preparation is between 90 and 150nm, and the encapsulation rate is between 90 and 98 percent. Different from the liposome reported in the prior literature and patents, the platinum liposome has the characteristics of high entrapment rate, good stability and strong anticancer activity.
EXAMPLE 38 preparation of reference liposomes
According to a literature method (CN106995465B), a platinum complex a (structure shown as a) is prepared from cisplatin and an amphiphilic compound b (structure shown as b) as a platinum complex containing amido bonds, and a liposome is prepared according to example 31 in the patent (CN106995465B), wherein the dosage of the platinum complex a is equivalent to 12mg of cisplatin, the dosage of a membrane material is 20mg of cholesterol, 200mg of soybean lecithin, the dosage of an antioxidant VE is 5mg, a hydration medium is water for injection, and the preparation method is a reverse evaporation method. The prepared liposome has the particle size of 101.7nm and the entrapment rate of 92.76%.
EXAMPLE 39 pharmaceutical Effect of liposomes
1. Inhibition rate of tumor
With normal saline, cisplatin for injection, the liposomes prepared in examples 32 and 33, and the reference liposome (the liposome prepared in example 38) as experimental materials, SPF-level KM mice, male, 6-8 weeks old, and 18-22 g in body weight were selected. The cell concentration was adjusted to (1-5). times.106 cells/mL using logarithmically growing sarcoma S180 cells. S180 cell suspension is inoculated to the right side of the mouse, the axilla is subcutaneous, each 0.2mL, and an S180 sarcoma strain axilla inoculation model is established. Tumor-bearing mice were randomly divided into 7 groups of 7 mice each, namely a physiological saline group, a cisplatin injection control group, an experimental group (to which the platinum complex liposome and the reference liposome provided in examples 32 and 33 of the present invention were administered, respectively), an intra-group label, a caudal vein of each group, a cisplatin control group, and an experimental group, and were administered once every two days, four times, once every three days, and four times at doses of 5mg/kg and 10mg/kg, respectively. After administration, the survival state of the mice was observed every day, the mice were sacrificed on the 20 th day after administration, tumors were taken out, the surfaces were washed with physiological saline, blotted with filter paper, weighed, and the tumor inhibition rate was calculated as follows:
tumor inhibition ratio (%) (average tumor weight in control group-average tumor weight in test group) ÷ average tumor weight in control group × 100
The experimental results show that the tumor growth of the administration group is slower than that of the control group, and compared with the cisplatin control group, the experimental group can obviously inhibit the tumor growth and can obviously prolong the survival time of tumor-bearing mice. The tumor weights and the related parameters are shown in Table 8, respectively, and the survival rates are shown in Table 9.
TABLE 8 Holosarcoma S180 tumor weight and tumor inhibition rate of 5mg/kg and 10mg/kg experimental material injected into KM mouse tail vein
Example number
Average tumor weight (g)
Inhibition ratio (%)
Physiological saline
4.5520
—
Example 32(5mg/kg)
1.7021
62.61
Example 32(10mg/kg)
0.6931
84.77
Example 33(5mg/kg)
1.7236
62.14
Example 33(10mg/kg)
0.7373
83.80
Reference liposome (5mg/kg)
1.7429
61.71
Reference liposome (10mg/kg)
0.7562
83.39
The results of KM mouse survival in Holland sarcoma S180 are shown in FIG. 7(7a-7d) and Table 9.
TABLE 9 Holoma S180 KM mice with 20-day survival rates
Example number
Survival rate (%)
Physiological saline
85.71
Cisplatin control group (5mg/kg)
0
Cisplatin control group (10mg/kg)
0
Example 32(5mg/kg)
85.71
Example 32(10mg/kg)
85.71
Example 33(5mg/kg)
85.71
Example 33(10mg/kg)
85.71
Reference liposome (5mg/kg)
71.43
Reference liposome (10mg/kg)
71.43
The results show that the tumor inhibition rate of the liposome prepared in example 32 is equivalent to that of a reference liposome with equal dose, and the survival rate is obviously higher than that of the reference liposome, which indicates that the liposome provided by the invention not only can effectively inhibit tumor growth and improve the survival rate of mice, but also obviously reduces the toxicity of cisplatin and improves the survival state of the mice, so that the toxicity of the platinum complex containing ester bonds is lower than that of the platinum complex containing amido bonds.
The results of KM mouse kidney tissue sections of he sarcoma S180 are shown in fig. 8.
The results show that the kidney of the CDDP-S group mice can see glomerular atrophy, renal vesicle expansion and obvious renal injury, but the liposome provided by the invention has no obvious renal injury, so that the liposome provided by the invention can obviously reduce the cisplatin nephrotoxicity.
EXAMPLE 40 Liposome safety
1. Survival State, body weight
KM mice were randomly grouped into groups of 10 mice each, labeled intra-group, fasted for 12h before administration, allowed to drink water freely, and treated with saline, a commercially available cisplatin control group, and the liposomes obtained in example 33 at a dose of 10mg/kg, respectively. Mice and body weight changes were observed and recorded for 14 consecutive days after dosing.
The results are shown in FIGS. 9 and 10. The results show that compared with the cisplatin control group, the liposome mouse prepared in the example 33 has the advantages of obviously reduced weight loss, good survival state and obviously improved survival rate, and the toxicity of the cisplatin complex liposome is obviously reduced.
2.LD50
KM mice were randomly divided into 14 dose groups of 10 mice each. After 1 day of group feeding, each group of mice was observed for normality, and the mice were weighed before administration. The dose of the intravenous injection of example 33 and the reference liposome was calculated for each mouse based on body weight, the group distance was 1.3, and the concentration of each dose group was 26, 33.8, 43.94, 57.12, 74.26, 96.54, 125.50 mg/kg. Within 7 days after dosing, the number of mice sacrificed in each group was recorded and during the experiment, the animals in each group were fed a free diet. The death number of the animal is input into LD50Value software, calculating LD50。
The results show that LD of the liposome prepared in example 3350LD value of 68.01mg/kg, reference liposome (Ex. 38)50The value was 45.90mg/kg, so the liposomes prepared in example 33 were safer than the reference liposomes. Further, it was confirmed that the platinum-based complex having an ester bond is less toxic than the platinum-based complex having an amide bond, and it is considered that the former is less toxic to metabolites in vivo than the latter, and therefore is more safe.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.