CN102284059A - Photosensitizer, as well as preparation method and application thereof - Google Patents

Abstract

The invention relates to a photosensitizer, as well and a preparation method and application thereof. The photosensitizer is a liposome preparation consisting of phthalocyanine zinc complex, which can be dispersed uniformly and exist stably for a long time in an aqueous solution, so that the photosensitizer can be used as an injection medicament conveniently, has higher target performance, and can better plays the drug action. Meanwhile, the preparation has the effects of killing and inhibiting various bacteria widely, and strong killing effect to various cancer cells, has the advantages of uniform shape, high entrapment rate and low permeability, and is in accordance with various requirements of pharmacopoeia for the liposome preparation.

Description

Translated fromChinese

一种光敏剂及其制备方法和应用A kind of photosensitizer and its preparation method and application

技术领域technical field

本发明涉及生物制药领域，更具体涉及由酞菁锌配合物组成的光敏剂。The invention relates to the field of biopharmaceuticals, in particular to a photosensitizer composed of zinc phthalocyanine complex.

背景技术Background technique

脂质体是由类脂分子层形成的，内部为水相的闭合囊泡，粒径从几十纳米到几十微米。脂质体具有类似细胞结构，有生物膜的特性和功能，它可以包裹水溶性和脂溶性药物，水溶性药物包裹于内部水相中，脂溶性药物插入于脂质双分子层，脂质体作为药物载体可以控制药物的释放，提高药物的靶向性，降低药物毒副作用、提高生物利用度、并具有长效缓释作用。1998年第一个脂质体制剂，即含益康唑的脂质体凝胶“Pevary Lipogel”在瑞士CILAG制药公司注册，并在瑞士、意大利、比利时和挪威等国上市销售，脂质体与生物膜亲和力强，能提高药物的粘膜透过性，如将青霉素脂质体制成眼用制剂时，局部透过角膜的能力是单纯药物的4倍，实验证明，只用原剂量的1/10即可具有角膜透过能力。此外，还有研究表明，脂质体能与细菌膜融合，直接使药物进入细胞浆，从而显著增加药物的抗菌能力。某些特殊的细胞如绿脓假单胞菌能产生磷脂酶，促使脂质体在细菌的感染部位释放高浓度药物。近年来脂质体局部给药的研究开展广泛，脂质体对创伤或病变皮肤、粘膜起治疗和保护作用，既有利于药物与患处接触，在局部发挥药效，又可避免因全身吸收而产生的不良反应。Liposomes are formed by a layer of lipid molecules, and the interior is a closed vesicle in the water phase, with a particle size ranging from tens of nanometers to tens of microns. Liposome has a similar cell structure and has the characteristics and functions of biological membranes. It can wrap water-soluble and fat-soluble drugs. Water-soluble drugs are wrapped in the internal water phase, and fat-soluble drugs are inserted into lipid bilayers. Liposomes As a drug carrier, it can control the release of drugs, improve the targeting of drugs, reduce the toxic and side effects of drugs, improve bioavailability, and have a long-term sustained release effect. In 1998, the first liposome preparation, the liposome gel "Pevary Lipogel" containing econazole, was registered at CILAG Pharmaceutical Company in Switzerland, and was marketed in Switzerland, Italy, Belgium, Norway and other countries. Liposome and Strong biomembrane affinity can improve the mucosal permeability of drugs. For example, when penicillin liposomes are made into ophthalmic preparations, the local penetrating ability of the cornea is 4 times that of simple drugs. Experiments have proved that only 1/10 of the original dose is used. It can have corneal penetration ability. In addition, studies have shown that liposomes can fuse with bacterial membranes and directly allow drugs to enter the cytoplasm, thereby significantly increasing the antibacterial ability of drugs. Certain special cells such as Pseudomonas aeruginosa can produce phospholipase, prompting liposomes to release high concentrations of drugs at the site of bacterial infection. In recent years, the research on liposome topical administration has been carried out extensively. Liposome has a therapeutic and protective effect on wounded or diseased skin and mucous membranes. Adverse reactions produced.

α-(8-喹啉氧基)单取代酞菁锌（简称：F7）已获得中国专利，专利号为200810070879.1，结构如图1所示。α-(8-quinolineoxy) monosubstituted zinc phthalocyanine (abbreviation: F7) has obtained a Chinese patent, the patent number is 200810070879.1, and the structure is shown in Figure 1.

F7表现出很高的光动力活性，其对胃癌BGC-823细胞、肝癌BEL7402细胞和肝癌HepG2细胞的半致死浓度IC₅₀(细胞死亡50％的浓度)分别为 0.02、0.024、0.08mmol/L，且F7对这几种肿瘤细胞的暗毒性很低。在光照时，其光灭活癌细胞能力很强，这是作为理想光敏剂必备的条件。F7对正常HELF细胞无论是光毒性和暗毒性都很低，而且都低于其对癌细胞相应的毒性。理想光敏剂应该对癌细胞具有高的光毒性和低的暗毒性，且对正常细胞具有较低的光毒性和暗毒性，这样才能在充分发挥光敏剂的光动力活性的同时又不损伤正常细胞，减少毒副作用。因此，以上研究结果表明F7是性能优良的光敏剂，有望开发成为光动力治疗用光敏剂。BEL7402癌细胞对F7的摄取率明显高于正常细胞。F7 showed high photodynamic activity, and its semi-lethal concentration_IC50 (50% cell death concentration) for gastric cancer BGC-823 cells, liver cancer BEL7402 cells and liver cancer HepG2 cells were 0.02, 0.024, 0.08mmol/L, respectively, And the dark toxicity of F7 to these kinds of tumor cells is very low. When exposed to light, it has a strong ability to photoinactivate cancer cells, which is a necessary condition for being an ideal photosensitizer. Both phototoxicity and dark toxicity of F7 to normal HELF cells are very low, and both are lower than its corresponding toxicity to cancer cells. An ideal photosensitizer should have high phototoxicity and low dark toxicity to cancer cells, and low phototoxicity and dark toxicity to normal cells, so that the photodynamic activity of the photosensitizer can be fully exerted without damaging normal cells. , reduce toxic side effects. Therefore, the above research results show that F7 is a photosensitizer with excellent performance, and it is expected to be developed as a photosensitizer for photodynamic therapy. The uptake rate of BEL7402 cancer cells to F7 was significantly higher than that of normal cells.

F7的水溶性极差，几乎不溶于水。这就给给药途径带来了极大困难，特别是静脉注射给药，该缺陷大大限制了F7在抗癌方面的使用范围。F7 has extremely poor water solubility and is almost insoluble in water. This brings great difficulties to the route of administration, especially intravenous administration, and this defect greatly limits the scope of use of F7 in anticancer.

发明内容Contents of the invention

为了克服以上问题，本发明提供了一种光敏剂及其制备方法和应用，该光敏剂是由酞菁锌配合物组成的脂质体制剂，能在水溶液中均匀分散且稳定存在，因而可以方便的作为注射药物使用。In order to overcome the above problems, the present invention provides a photosensitizer and its preparation method and application. The photosensitizer is a liposome preparation composed of a phthalocyanine zinc complex, which can be uniformly dispersed in an aqueous solution and exists stably, so it can be conveniently used as an injectable drug.

本发明是通过如下技术方案实施的：The present invention is implemented through the following technical solutions:

一种光敏剂是由酞菁锌配合物组成的脂质体制剂。A photosensitizer is a liposomal formulation composed of a zinc phthalocyanine complex.

所述光敏剂的原料包括酞菁锌配合物、磷脂、亲脂性化合物、稳定剂、有机溶剂和水。The raw materials of the photosensitizer include zinc phthalocyanine complex, phospholipid, lipophilic compound, stabilizer, organic solvent and water.

所述酞菁锌配合物的摩尔浓度为1.48-104.00mmol/L，磷脂的浓度为0.1-10g/L，磷脂与亲脂性化合物质量比为10:1-1:2，稳定剂的质量浓度为0.01%-2%，有机溶剂体积与水的体积比为1:20-10:1。The molar concentration of the zinc phthalocyanine complex is 1.48-104.00mmol/L, the concentration of the phospholipid is 0.1-10g/L, the mass ratio of the phospholipid to the lipophilic compound is 10:1-1:2, and the mass concentration of the stabilizer is 0.01%-2%, the volume ratio of organic solvent to water is 1:20-10:1.

所述酞菁锌配合物选自α-（8-喹啉氧基）单取代酞菁锌、α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌、β-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌、双α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌、α-喹啉-低聚乙二醇酞菁锌和β-喹啉-低聚乙二醇酞菁锌和二-α-喹啉-低聚乙二醇酞菁锌。The zinc phthalocyanine complex is selected from α-(8-quinolineoxy) monosubstituted zinc phthalocyanine, α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6- Dioxa-1-octyloxy)zinc phthalocyanine, β-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)phthalocyanine Zinc cyanine, bis-α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)zinc phthalocyanine, α-quinoline-oligomeric Zinc ethylene glycol phthalocyanine and β-quinoline-oligoethylene glycol phthalocyanine zinc and di-α-quinoline-oligoethylene glycol phthalocyanine zinc.

所述磷脂选自二硬脂酰磷脂酰胆碱、二棕榈酰磷脂胆酰、二肉豆蔻酰磷脂酰胆碱、卵磷脂、大豆磷脂、二肉豆蔻酰磷脂酰甘油、2,2-二羟甲基丙酸、磷脂酰甘油、叠氮磷酸二苯酯、饱和卵磷脂、二硬脂酰磷脂酰甘油、氢化大豆卵磷脂、磷脂酰丝氨酸和鞘磷脂。The phospholipids are selected from the group consisting of distearoylphosphatidylcholine, dipalmitoylphosphatidylcholine, dimyristoylphosphatidylcholine, lecithin, soybean lecithin, dimyristoylphosphatidylglycerol, 2,2-dihydroxy Methylpropionic acid, phosphatidylglycerol, diphenylphosphoryl azide, saturated lecithin, distearoylphosphatidylglycerol, hydrogenated soy lecithin, phosphatidylserine, and sphingomyelin.

所述的亲脂性化合物选自C₁₂-C₁₈直连饱和或不饱和脂肪酸、单甘脂、生育酚和胆固醇。The lipophilic compound is selected from C₁₂ -C₁₈ straight saturated or unsaturated fatty acids, monoglycerides, tocopherol and cholesterol.

所述稳定剂选自甘露醇、葡萄糖、聚乙二醇、泊洛沙姆、吐温、司盘和右旋糖苷。The stabilizer is selected from the group consisting of mannitol, glucose, polyethylene glycol, poloxamer, Tween, span and dextran.

所述有机溶剂选自THF、乙醇、乙醚、氯仿和三氯甲烷。The organic solvent is selected from THF, ethanol, ether, chloroform and chloroform.

所述制备方法包括以下步骤：按所述比例将磷脂，胆固醇，酞菁锌配合物和稳定剂用有机溶剂溶解后，将所得溶液注入到蒸馏水中，其有机溶剂与蒸馏水体积比为1:20-10:1，边加边搅拌，加完后在温度10-70℃条件下减压蒸发除去有机溶剂，得到单取代酞箐锌脂质体制剂，即所述的光敏剂。The preparation method comprises the following steps: after dissolving the phospholipid, cholesterol, zinc phthalocyanine complex and stabilizer in an organic solvent according to the stated ratio, injecting the obtained solution into distilled water, and the volume ratio of the organic solvent to the distilled water is 1:20 -10:1, stirring while adding, after the addition is completed, the organic solvent is evaporated under reduced pressure at a temperature of 10-70°C to obtain a monosubstituted zinc phthalocyanine liposome preparation, that is, the photosensitizer.

该制备方法得到的脂质体的粒径为50nm-5微米。The particle size of the liposome obtained by the preparation method is 50nm-5 microns.

所述应用是该光敏剂作为一种光动力治疗药物使用。The application is that the photosensitizer is used as a photodynamic therapy drug.

本发明的脂质体制剂的制备方法选自薄膜法、逆向蒸发法、熔融法、注入法、冷冻干燥法、表面活性剂法、复乳法和钙融合法。但优选注入法，即本发明所述的方法。The preparation method of the liposome preparation of the present invention is selected from thin film method, reverse evaporation method, melting method, injection method, freeze-drying method, surfactant method, double emulsion method and calcium fusion method. However, the injection method is preferred, ie the method described in the present invention.

由α-（8-喹啉氧基）单取代酞箐锌组成的脂质体，简称：W7。Liposome composed of α-(8-quinolinyloxy) monosubstituted zinc phthalocyanine, referred to as: W7.

本发明的优点为：The advantages of the present invention are:

1）本发明制备的酞菁锌配合物组成的脂质体制剂是一种第三代光敏剂，因此其可以在水溶液中均匀分散且较长时间稳定存在，且具有较高的靶向性，能更好的发挥药效，因而可以方便的作为注射药物使用，更好的发挥药效。1) The liposome preparation composed of the phthalocyanine zinc complex prepared in the present invention is a third-generation photosensitizer, so it can be uniformly dispersed in aqueous solution and exist stably for a long time, and has high targeting, The medicine effect can be better exerted, so it can be conveniently used as an injection medicine, and the medicine effect can be better exerted.

2）本发明制备的脂质体制剂，可作为临床治疗的药物制剂，用于抗癌及杀菌的治疗。该制剂使用方便，疗效显著，安全可靠，对各类细菌具有广泛的灭杀和抑制作用，对各种癌细胞具有很强的杀伤作用。2) The liposome preparation prepared by the present invention can be used as a pharmaceutical preparation for clinical treatment, for anticancer and bactericidal treatment. The preparation is convenient to use, has remarkable curative effect, is safe and reliable, has extensive killing and inhibiting effects on various bacteria, and has strong killing effects on various cancer cells.

3）本发明制备的脂质体制剂的质量安全可靠，根据中华人民共和国药典2005年版二部，对脂质体的质量进行检查。结果表明，该方法制备的脂质体平均粒径为50nm-5微米，分散系数为0.01-0.60。形态均一，包封率高，渗透率低，符合药典对脂质体制剂的各项要求。3) The quality of the liposome preparation prepared by the present invention is safe and reliable, and the quality of the liposome is checked according to the second part of the Pharmacopoeia of the People's Republic of China in 2005. The results show that the average particle size of the liposome prepared by the method is 50nm-5 microns, and the dispersion coefficient is 0.01-0.60. Uniform shape, high encapsulation rate, low permeability, in line with the requirements of the Pharmacopoeia for liposome preparations.

附图说明Description of drawings

图1为α-(8-喹啉氧基)单取代酞菁锌的分子结构式；Fig. 1 is the molecular structural formula of α-(8-quinoline oxygen group) monosubstituted zinc phthalocyanine;

图2为α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌；Fig. 2 is α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)zinc phthalocyanine;

图3为β-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌；Fig. 3 is β-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)zinc phthalocyanine;

图4为双α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌；Fig. 4 is double α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)zinc phthalocyanine;

图5为α-喹啉-低聚乙二醇酞菁锌，其中n=2、3、4、5、6、8、9、28；Fig. 5 is α-quinoline-oligoethylene glycol phthalocyanine zinc, wherein n=2, 3, 4, 5, 6, 8, 9, 28;

图6为β-喹啉-低聚乙二醇酞菁锌，其中n=2、3、4、5、6、8、9、28；Fig. 6 is β-quinoline-oligoethylene glycol phthalocyanine zinc, wherein n=2, 3, 4, 5, 6, 8, 9, 28;

图7为二-α-喹啉-低聚乙二醇酞菁锌，其中n=2、3、4、5、6、8、9、28；Fig. 7 is two-alpha-quinoline-oligoethylene glycol phthalocyanine zinc, wherein n=2, 3, 4, 5, 6, 8, 9, 28;

图8为W7的UV-Vis光谱图；Figure 8 is the UV-Vis spectrum of W7;

图9为W7对胃癌BGC823细胞的光毒暗毒性测试；Figure 9 shows the phototoxicity and dark toxicity test of W7 on gastric cancer BGC823 cells;

图10为W7对肝癌HepG2细胞的光毒暗毒性测试；Figure 10 shows the phototoxicity and dark toxicity test of W7 to liver cancer HepG2 cells;

图11为W7对正常细胞HepG2细胞的光毒暗毒性。Figure 11 shows the phototoxicity and dark toxicity of W7 to normal cells HepG2 cells.

具体实施方式Detailed ways

一种光敏剂的原料包括酞菁锌配合物、磷脂、亲脂性化合物、稳定剂和水。A raw material for a photosensitizer includes a zinc phthalocyanine complex, a phospholipid, a lipophilic compound, a stabilizer and water.

所述酞菁锌配合物的摩尔浓度为1.48-104.00微mol/L，磷脂的浓度为0.1-10g/L，磷脂与亲脂性化合物质量比为10:1-1:2，稳定剂的质量浓度为0.01%-2%，有机溶剂体积与水的体积比为1:20-10:1。The molar concentration of the zinc phthalocyanine complex is 1.48-104.00 micromol/L, the concentration of the phospholipid is 0.1-10g/L, the mass ratio of the phospholipid to the lipophilic compound is 10:1-1:2, and the mass concentration of the stabilizer is 0.01%-2%, the volume ratio of organic solvent to water is 1:20-10:1.

所述磷脂选自二硬脂酰磷脂酰胆碱、二棕榈酰磷脂胆酰、二肉豆蔻酰磷脂酰胆碱、卵磷脂、大豆磷脂、二肉豆蔻酰磷脂酰甘油、2,2-二羟甲基丙酸、磷脂酰甘油、叠氮磷酸二苯酯、饱和卵磷脂、二硬脂酰磷脂酰甘油、氢化大豆卵磷脂、磷脂酰丝氨酸和鞘磷脂。The phospholipids are selected from the group consisting of distearoylphosphatidylcholine, dipalmitoylphosphatidylcholine, dimyristoylphosphatidylcholine, lecithin, soybean lecithin, dimyristoylphosphatidylglycerol, 2,2-dihydroxy Methylpropionic acid, phosphatidylglycerol, diphenylphosphoryl azide, saturated lecithin, distearoylphosphatidylglycerol, hydrogenated soy lecithin, phosphatidylserine, and sphingomyelin.

所述的亲脂性化合物选自C₁₂-C₁₈直连饱和或不饱和脂肪酸、单甘脂、生育酚和胆固醇。The lipophilic compound is selected from C₁₂ -C₁₈ straight saturated or unsaturated fatty acids, monoglycerides, tocopherol and cholesterol.

所述稳定剂选自甘露醇、葡萄糖、聚乙二醇、泊洛沙姆、吐温、司盘和右旋糖苷。The stabilizer is selected from the group consisting of mannitol, glucose, polyethylene glycol, poloxamer, Tween, span and dextran.

所述有机溶剂选自THF、乙醇、乙醚、氯仿和三氯甲烷。The organic solvent is selected from THF, ethanol, ether, chloroform and chloroform.

所述制备方法包括以下步骤：按所述比例将磷脂，胆固醇，酞菁锌配合物和稳定剂用有机溶剂溶解后，将所得溶液注入到蒸馏水中，其有机溶剂与蒸馏水体积比为1:20-10:1，边加边搅拌，加完后在温度10-70℃条件下减压蒸发除去THF，得到酞菁锌配合物脂质体制剂，即所述的光敏剂。The preparation method comprises the following steps: after dissolving the phospholipid, cholesterol, zinc phthalocyanine complex and stabilizer in an organic solvent according to the stated ratio, injecting the obtained solution into distilled water, and the volume ratio of the organic solvent to the distilled water is 1:20 -10:1, stirring while adding, after the addition, evaporate and remove THF under reduced pressure at a temperature of 10-70°C to obtain a zinc phthalocyanine complex liposome preparation, that is, the photosensitizer.

该制备方法得到的脂质体的粒径为50nm-5微米。The particle size of the liposome obtained by the preparation method is 50nm-5 microns.

所述应用是该光敏剂作为一种光动力治疗药物使用。The application is that the photosensitizer is used as a photodynamic therapy drug.

实施例1Example 1

一种光敏剂的原料包括α-（8-喹啉氧基）单取代酞菁锌、磷脂、亲脂性化合物、稳定剂、有机溶剂和水。A raw material of a photosensitizer includes α-(8-quinolinyloxy) monosubstituted zinc phthalocyanine, phospholipid, lipophilic compound, stabilizer, organic solvent and water.

所述α-（8-喹啉氧基）单取代酞箐锌的摩尔浓度为1.48微mol/L，磷脂的浓度为0.1g/L，磷脂与亲脂性化合物质量比为10:1，稳定剂的质量浓度为0.01%，有机溶剂体积与水的体积比为1:20-10:1。The molar concentration of the α-(8-quinolineoxy) monosubstituted zinc phthalocyanine is 1.48 micromol/L, the concentration of phospholipid is 0.1 g/L, the mass ratio of phospholipid to lipophilic compound is 10:1, and the stabilizer The mass concentration is 0.01%, and the volume ratio of organic solvent to water is 1:20-10:1.

所述磷脂选自二硬脂酰磷脂酰胆碱和二棕榈酰磷脂胆酰。The phospholipids are selected from distearoylphosphatidylcholine and dipalmitoylphosphatidylcholine.

所述的亲脂性化合物选自生育酚和胆固醇。Said lipophilic compound is selected from tocopherol and cholesterol.

所述稳定剂选自司盘和右旋糖苷。The stabilizer is selected from Span and dextran.

所述有机溶剂选自THF。The organic solvent is selected from THF.

所述制备方法包括以下步骤：按所述比例将磷脂，胆固醇，α-（8-喹啉氧基）单取代酞箐锌和稳定剂用有机溶剂溶解后，将所得溶液注入到蒸馏水中，其有机溶剂与蒸馏水体积比为1:20，边加边搅拌，加完后在温度10-70℃条件下减压蒸发除去有机溶剂，得到α-（8-喹啉氧基）单取代酞箐锌脂质体制剂，即所述的第三代光敏剂。The preparation method comprises the following steps: after dissolving phospholipid, cholesterol, α-(8-quinolineoxy) monosubstituted zinc phthalocyanine and stabilizer in an organic solvent according to the stated ratio, injecting the obtained solution into distilled water, and The volume ratio of organic solvent to distilled water is 1:20. Stir while adding. After the addition, evaporate under reduced pressure at a temperature of 10-70°C to remove the organic solvent to obtain α-(8-quinolineoxy) monosubstituted zinc phthalocyanine Liposome preparation, that is, the third-generation photosensitizer.

实施例2Example 2

一种第三代光敏剂的原料包括α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌、磷脂、亲脂性化合物、稳定剂、有机溶剂和水，有机溶剂体积与水的体积比为10:1。A third-generation photosensitizer raw material includes α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)zinc phthalocyanine, phospholipid , lipophilic compound, stabilizer, organic solvent and water, the volume ratio of organic solvent volume to water is 10:1.

所述α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌的摩尔浓度为104.00微mol/L，磷脂的浓度为10g/L，磷脂与亲脂性化合物质量比为1:2，稳定剂的质量浓度为2%。The molar concentration of the α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)zinc phthalocyanine is 104.00 micromol/L, The concentration of phospholipids is 10g/L, the mass ratio of phospholipids to lipophilic compounds is 1:2, and the mass concentration of stabilizers is 2%.

所述磷脂为磷脂酰甘油。The phospholipid is phosphatidylglycerol.

所述的亲脂性化合物为C₁₂-C₁₈直连饱和脂肪酸。The lipophilic compound is C₁₂ -C₁₈ straight-linked saturated fatty acid.

所述稳定剂为甘露醇。The stabilizer is mannitol.

所述有机溶剂选自氯仿。The organic solvent is selected from chloroform.

所述制备方法包括以下步骤：按所述比例将磷脂，胆固醇，α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌和稳定剂用有机溶剂溶解后，将所得溶液注入到蒸馏水中，其有机溶剂与蒸馏水体积比为10:1，边加边搅拌，加完后在温度10-70℃条件下减压蒸发除去有机溶剂，得到α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌脂质体制剂，即所述的第三代光敏剂。The preparation method comprises the following steps: phospholipid, cholesterol, α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy Base) after dissolving zinc phthalocyanine and stabilizer with organic solvent, inject the obtained solution into distilled water, the volume ratio of organic solvent to distilled water is 10:1, stir while adding, after adding, under the condition of temperature 10-70°C The organic solvent was removed by evaporation under reduced pressure to obtain α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy) phthalocyanine zinc liposome preparation , the third generation photosensitizer.

实施例3Example 3

一种第三代光敏剂的原料包括双α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌、磷脂、亲脂性化合物、稳定剂、有机溶剂和水，有机溶剂体积与水的体积比为1: 1。The raw material of a third-generation photosensitizer includes bis-α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)zinc phthalocyanine, Phospholipid, lipophilic compound, stabilizer, organic solvent and water, the volume ratio of organic solvent volume and water is 1: 1.

所述双α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌的摩尔浓度为50mmol/L，磷脂的浓度为1g/L，磷脂与亲脂性化合物质量比为1:1，稳定剂的质量浓度为1%。The molar concentration of the two α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)zinc phthalocyanine is 50mmol/L, and the phospholipid The concentration is 1g/L, the mass ratio of phospholipid to lipophilic compound is 1:1, and the mass concentration of stabilizer is 1%.

所述磷脂为叠氮磷酸二苯酯。The phospholipid is diphenylphosphoryl azide.

所述的亲脂性化合物为单甘脂。The lipophilic compound is monoglyceride.

所述稳定剂为聚乙二醇。Described stabilizing agent is polyethylene glycol.

所述有机溶剂选自氯仿和三氯甲烷。The organic solvent is selected from chloroform and chloroform.

所述制备方法包括以下步骤：按所述比例将磷脂，胆固醇，双α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌和稳定剂用有机溶剂溶解后，将所得溶液注入到蒸馏水中，其有机溶剂与蒸馏水体积比为1:1，边加边搅拌，加完后在温度10-70℃条件下减压蒸发除去有机溶剂，得到双α-(8-(4，6-二甲基-2-嘧啶氧基)-3，6-二氧杂-1-辛氧基)酞菁锌脂质体制剂，即所述的第三代光敏剂。The preparation method comprises the following steps: mixing phospholipid, cholesterol, bis-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyl After dissolving the oxy)zinc phthalocyanine and the stabilizer in an organic solvent, inject the resulting solution into distilled water, the volume ratio of the organic solvent to the distilled water is 1:1, stir while adding, and keep the temperature at 10-70°C after adding The organic solvent was evaporated under reduced pressure to obtain bis-α-(8-(4,6-dimethyl-2-pyrimidinyloxy)-3,6-dioxa-1-octyloxy)zinc phthalocyanine lipid Body preparation, that is, the third generation photosensitizer.

性能测试Performance Testing

W7的光降解速率常数为0.0039 min^-1，相同条件下F7的光降解速率常数为0.014min^-1。说明W7比F7更稳定。The photodegradation rate constant of W7 was 0.0039 min^-1 , and the photodegradation rate constant of F7 was 0.014 min^-1 under the same conditions. It shows that W7 is more stable than F7.

W7光敏化产生¹O₂速率常数为0.0114 min^-1，相同条件下F7光敏化产生¹O₂速率常数为0.0035 min^-1。说明W7产生¹O₂的能力强于F7。The rate constant of¹ O₂ produced by photosensitization of W7 was 0.0114 min^-1 , and the rate constant of¹ O₂ produced by photosensitization of F7 was 0.0035 min^-1 under the same conditions. It shows that the ability of W7 to produce¹ O₂ is stronger than that of F7.

W7光敏氧化酪氨酸的速率常数为0.024 min^-1，相同条件下F7光敏化酪氨酸的速率常数为0.008 min^-1。说明W7光敏氧化酪氨酸的能力强于F7。The rate constant of W7 photosensitized tyrosine oxidation was 0.024 min^-1 , and the rate constant of F7 photosensitized tyrosine was 0.008 min^-1 under the same conditions. It shows that the ability of W7 to photosensitively oxidize tyrosine is stronger than that of F7.

由上述结果可知在光化学性质方面，相对于F7，W7表现出作为光敏药物的明显优势。From the above results, it can be seen that in terms of photochemical properties, compared with F7, W7 has obvious advantages as a photosensitive drug.

同时，由图3，图4可知，W7对胃癌BGC823细胞、肝癌HepG2细胞具有很强的杀伤作用，其IC50值都很低（分别为1.42μmol/L、1.46μmol/L）。At the same time, it can be seen from Figure 3 and Figure 4 that W7 has a strong killing effect on gastric cancer BGC823 cells and liver cancer HepG2 cells, and its IC50 values are very low (1.42 μmol/L and 1.46 μmol/L, respectively).

由图5可知，W7对正常细胞HepG2细胞的光毒性暗毒性都比较低。在较高浓度范围内W7对正常细胞HepG2细胞都几乎没有光毒性和暗毒性。It can be seen from Figure 5 that the phototoxicity and dark toxicity of W7 to normal cells HepG2 cells are relatively low. In a higher concentration range, W7 has almost no phototoxicity and dark toxicity to normal cells HepG2 cells.

因此，W7具有很好的选择性杀伤癌细胞。Therefore, W7 has good selective killing of cancer cells.

以上所述仅为本发明的较佳实施例，凡依本发明申请专利范围所做的均等变化与修饰，皆应属本发明的涵盖范围。The above descriptions are only preferred embodiments of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention shall fall within the scope of the present invention.

Claims (10)

1. photosensitizer is characterized in that: the Liposomal formulation that described photosensitizer is made up of phthalocyanine Zn complex, its particle diameter are 50 nanometers-5 micron.

2. photosensitizer according to claim 1 is characterized in that: the raw material of described photosensitizer comprises phthalocyanine Zn complex, phospholipid, lipophilic compound, stabilizing agent, organic solvent and water.

3. a kind of photosensitizer according to claim 1 and 2, it is characterized in that: the molar concentration of described phthalocyanine Zn complex is the little mol/L of 1.48-104.00, the concentration of phospholipid is 0.1-10g/L, phospholipid and lipophilic compound mass ratio are 10:1-1:2, the mass concentration of stabilizing agent is 0.01%-2%, and the volume ratio of organic solvent volume and water is 1:20-10:1.

4. photosensitizer according to claim 2, it is characterized in that: described phthalocyanine Zn complex is selected from α-(8-quinoline oxy) mono-substituted zinc phthalocyanine, α-(8-(4,6-dimethyl-2-2-pyrimidinyl oxy)-3,6-two oxa-s-1-octyloxy) Phthalocyanine Zinc, β-(8-(4,6-dimethyl-2-2-pyrimidinyl oxy)-3,6-two oxa-s-1-octyloxy) Phthalocyanine Zinc, two α-(8-(4,6-dimethyl-2-2-pyrimidinyl oxy)-3,6-two oxa-s-1-octyloxy) Phthalocyanine Zinc, α-quinoline-low Polyethylene Glycol Phthalocyanine Zinc and β-quinoline-low Polyethylene Glycol Phthalocyanine Zinc and two-α-quinoline-low Polyethylene Glycol Phthalocyanine Zinc.

5. a kind of photosensitizer according to claim 2, it is characterized in that: described phospholipid is selected from distearoyl phosphatidylcholine, two palmityl phospholipid gallbladder acyls, dimyristoyl phosphatidyl choline, lecithin, soybean phospholipid, GLYCEROL,DIMYRISTOYL PHOSPHATIDYL, 2,2-dihydromethyl propionic acid, phosphatidyl glycerol, diphenyl phosphate azide, saturated lecithin, distearyl phosphatidyl glycerol, hydrogenated soy phosphatidyl choline, Phosphatidylserine and sphingomyelins.

6. a kind of photosensitizer according to claim 2 is characterized in that: described lipophilic compound is selected from C₁₂-C₁₈Direct-connected saturated or unsaturated fatty acid, monoglyceride, tocopherol and cholesterol.

7. a kind of photosensitizer according to claim 2 is characterized in that: described stabilizing agent is selected from mannitol, glucose, Polyethylene Glycol, poloxamer, tween, span and dextran.

8. a kind of photosensitizer according to claim 2 is characterized in that: described organic solvent is selected from THF, ethanol, ether, chloroform and chloroform.

9. preparation method as claim 1 or 3 described photosensitizer, it is characterized in that: described preparation method may further comprise the steps: in described ratio with phospholipid, cholesterol, phthalocyanine Zn complex and stabilizing agent with organic solvent dissolution after, gained solution is injected in the distilled water, and its organic solvent and distilled water volume ratio are 1:20-10:1, and the limit edged stirs, add back reduction vaporization under temperature 10-70 ℃ condition and remove organic solvent, obtain described photosensitizer.

10. the application of the photosensitizer of a photosensitizer as claimed in claim 1 or preparation method as claimed in claim 9 preparation, it is characterized in that: described application is that this photosensitizer uses as a kind of optical dynamic therapy medicine.

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