WO2018113094A1

WO2018113094A1 - Use of fullerene structure in preparation of drug for treating diabetes and complications thereof

Info

Publication number: WO2018113094A1
Application number: PCT/CN2017/075445
Authority: WO
Inventors: 王春儒; 甄明明; 李雪; ***; 李慧
Original assignee: 北京福纳康生物技术有限公司
Priority date: 2016-12-19
Filing date: 2017-03-02
Publication date: 2018-06-28

Abstract

Disclosed is the use of a fullerene structure in the preparation of a drug for treating diabetes and complications thereof. The fullerene structure comprises at least one effective ingredient selected from the group consisting of an oil-soluble fullerene, an oil-soluble endohedral metallofullerene, a composition of the oil-soluble fullerene and the oil-soluble endohedral metallofullerene, a water-soluble fullerene, a water-soluble endohedral metallofullerene, a composition of the water-soluble fullerene and the water-soluble endohedral metallofullerene, pharmaceutically acceptable esters of the above six ingredients or pharmaceutically acceptable salts of the above six ingredients.

Description

富勒烯结构在制备治疗糖尿病及其并发症的药物中的应用Application of fullerene structure in the preparation of drugs for treating diabetes and its complications

交叉引用cross reference

本发明要求北京福纳康生物技术有限公司和中国科学院化学研究所向中国专利局提交的、申请号为CN201611180065.4、发明名称为“油溶性富勒烯在制备治疗糖尿病并发症的药物中的应用”的中国专利申请的优先权，该申请的全部内容通过引用结合在本发明中。The invention claims that Beijing Funakan Biotechnology Co., Ltd. and the Institute of Chemistry of the Chinese Academy of Sciences submitted to the Chinese Patent Office, the application number is CN201611180065.4, and the invention name is "oil-soluble fullerenes in the preparation of drugs for treating diabetic complications". The priority of the Chinese Patent Application, the entire disclosure of which is incorporated herein by reference.

本发明还要求中国科学院化学研究所和北京福纳康生物技术有限公司向中国专利局提交的、申请号为CN201611179710.0、发明名称为“水溶性富勒烯结构在制备治疗糖尿病的药物中的应用”的中国专利申请的优先权，该申请的全部内容通过引用结合在本发明中。The invention also requires the Chinese Academy of Sciences Institute of Chemistry and Beijing Funakan Biotechnology Co., Ltd. to submit to the Chinese Patent Office, the application number is CN201611179710.0, and the invention name is "water-soluble fullerene structure in the preparation of drugs for treating diabetes. The priority of the Chinese Patent Application, the entire disclosure of which is incorporated herein by reference.

本发明还要求中国科学院化学研究所和北京福纳康生物技术有限公司向中国专利局提交的、申请号为CN 201611180063.5、发明名称为“富勒烯结构在制备增强胰岛素敏感性的药物中的应用”的中国专利申请的优先权，该申请的全部内容通过引用结合在本发明中。The invention also requires the application of the Chinese Academy of Sciences and Beijing Funakan Biotechnology Co., Ltd. to the Chinese Patent Office, the application number is CN 201611180063.5, and the invention name is "fullerene structure in the preparation of drugs for enhancing insulin sensitivity". The priority of the Chinese Patent Application, the entire contents of which is incorporated herein by reference.

技术领域Technical field

本发明属于医药领域，涉及富勒烯结构在制备治疗糖尿病及其并发症的药物中的应用，特别涉及油溶性富勒烯结构或水溶性富勒烯结构在制备治疗糖尿病及其并发症的药物中的应用。The invention belongs to the field of medicine and relates to the application of fullerene structure in preparing medicines for treating diabetes and its complications, in particular to oil-soluble fullerene structure or water-soluble fullerene structure in preparing medicine for treating diabetes and its complications. Application in .

背景技术Background technique

胰岛素是由胰岛β细胞受内源性或外源性物质，如：葡萄糖、乳糖、核糖、精氨酸、胰高血糖素等的刺激而分泌的一种蛋白质激素。胰岛素是机体内唯一降低血糖的激素，其同时能促进糖原、脂肪、蛋白质合成。糖尿病(DM)是一种以高血糖为特征的代谢性疾病，其主要是由于胰岛素分泌缺陷、胰岛素生物活性受损中的至少一种引起的糖类、蛋白质、脂肪代谢紊乱。糖尿病分为一型糖尿病、二型糖尿病和妊娠期糖尿病，一型糖尿病主要发于青少年，是胰岛素依赖型的糖尿病；二型糖尿病属于成人发病型糖尿病，多在35-40岁之后发作，占糖尿病患者的90％以上，其主要特点是胰岛素相对不足或胰岛素抵抗，其危害程度也最大。Insulin is a protein hormone secreted by islet beta cells stimulated by endogenous or exogenous substances such as glucose, lactose, ribose, arginine, glucagon, and the like. Insulin is the only hormone in the body that lowers blood sugar, and it also promotes the synthesis of glycogen, fat and protein. Diabetes (DM) is a metabolic disease characterized by hyperglycemia, which is mainly caused by saccharide, protein, and fat metabolism disorders caused by at least one of insulin secretion deficiency and impaired insulin biological activity. Diabetes is divided into type 1 diabetes, type 2 diabetes, and gestational diabetes. Type 1 diabetes is mainly caused by adolescents. It is insulin-dependent diabetes; type 2 diabetes is adult-onset diabetes, which occurs after 35-40 years old, accounting for more than 90% of diabetic patients. Its main feature is relatively insufficient insulin or insulin resistance, and its degree of harm is also the greatest.

长期血糖增高对人类生命健康安全造成极大的伤害，如：会导致大血管、微血管受损并危及心、脑、肾、周围神经、眼睛、足等，从而引起各种糖尿病并发症，如糖尿病肾病、视网膜病变、糖尿病神经病变、糖尿病足病、糖尿病皮肤病变、动脉粥样硬化及其引起的冠心病、脑卒中、下肢疼痛间歇跛行等等。据世界卫生组织统计，糖尿病并发症多达100多种，是迄今为止已知并发症最多的一种疾病。糖尿病死亡者中，有50％以上是心脑血管所致，10％是肾病变所致，并且因糖尿病截肢的患者是非糖尿病的10～20倍。据临床数据显示，糖尿病发病后的将近10年，就会有30％～40％的糖尿病患者至少发生一种并发症，并且并发症一旦产生，现有的药物治疗很难逆转，因此，糖尿病并发症的预防和治疗是一个巨大的难题，而治疗糖尿病并发症的药物研究也是特别急切且有意义的。Long-term blood sugar increases cause great harm to human life health and safety, such as: it can cause damage to large blood vessels and micro-vessels and endanger the heart, brain, kidney, peripheral nerves, eyes, feet, etc., causing various complications of diabetes, such as diabetes. Nephropathy, retinopathy, diabetic neuropathy, diabetic foot disease, diabetic skin lesions, atherosclerosis and its associated coronary heart disease, stroke, intermittent pain in lower extremities, and so on. According to the World Health Organization, there are more than 100 complications of diabetes, which is the most known complication to date. More than 50% of diabetic deaths are caused by cardiovascular and cerebrovascular diseases, 10% are caused by nephropathy, and patients with diabetes amputation are 10 to 20 times more likely to be non-diabetic. According to clinical data, in the past 10 years after the onset of diabetes, 30% to 40% of diabetic patients have at least one complication, and once the complications are generated, the existing medical treatment is difficult to reverse. Therefore, diabetes is complicated. Prevention and treatment of the disease is a huge problem, and drug research for the treatment of diabetic complications is particularly urgent and meaningful.

胰岛素抵抗(IR)是指胰岛素作用的靶器官对胰岛素作用的敏感性下降，即正常剂量的胰岛素产生低于正常生物学效应的一种状态，其具体表现为：机体组织对胰岛素无法做出相应的应答，胰岛素受体失活，葡萄糖利用的细胞无法识别胰岛素，导致血糖不能降低，胰岛素敏感性下降。1998年7月WHO将胰岛素抵抗综合征定义为：①胰岛素抵抗；②糖耐量异常；③血压≥160/90mmHg；④甘油三酯≥1.7mmol/L；⑤向心性肥胖；⑥体重指数BMI>30kg/m2；⑦腰臀比，男性>0.9，女性>0.85；⑧高尿酸血症；⑨微量白蛋白尿。一个个体存在糖尿病或糖耐量减退或胰岛素抵抗，并同时具有以上9项中的2项以上组合时，可定义为胰岛素抵抗综合征。胰岛素抵抗是2型糖尿病的主要病理特征，与许多的糖尿病并发症相关，如糖尿病肾病，眼病，足病，心血管病等。研究发现，胰岛素抵抗与这些疾病息息相关，改善胰岛素抵抗，增加胰岛素的敏感性对这些疾病的治疗有巨大的意义。Insulin resistance (IR) refers to a decrease in the sensitivity of the target organ of insulin action to insulin action, that is, a state in which a normal dose of insulin produces a lower than normal biological effect, which is manifested by the fact that the body tissue cannot respond to insulin. In response to the inactivation of the insulin receptor, glucose-utilized cells are unable to recognize insulin, resulting in a decrease in blood glucose and a decrease in insulin sensitivity. In July 1998, WHO defined insulin resistance syndrome as: 1 insulin resistance; 2 impaired glucose tolerance; 3 blood pressure ≥ 160/90 mmHg; 4 triglycerides ≥ 1.7 mmol / L; 5 centripetal obesity; 6 body mass index BMI > 30 kg /m2; 7 waist to hip ratio, male > 0.9, female > 0.85; 8 hyperuricemia; 9 microalbuminuria. An individual with diabetes or impaired glucose tolerance or insulin resistance, and at the same time having more than two of the above nine items, can be defined as insulin resistance syndrome. Insulin resistance is the main pathological feature of type 2 diabetes and is associated with many diabetic complications, such as diabetic nephropathy, eye disease, podiatric disease, and cardiovascular disease. The study found that insulin resistance is closely related to these diseases, improving insulin resistance and increasing insulin sensitivity have great significance for the treatment of these diseases.

富勒烯是除石墨、金刚石和无定型碳之外碳元素的另一种同素异形体。这类物质指的是由碳原子组成的笼状结构，其含量最多的分子是C₆₀，然后是C₇₀、C₈₄，其次是含量相对较少的C₇₆、C₇₈、C₈₂等。另外由于富勒烯的碳笼内部为空腔结构，因此其内部空腔可内嵌不同原子、离子或原子簇，被称之为内嵌富勒烯，如La@C₆₀，表示La内嵌在C₆₀的笼状结构中，@表示at，形象的表达了内嵌的含义。Fullerenes are another allotrope of carbon other than graphite, diamond and amorphous carbon. This type of substance refers to a cage structure composed of carbon atoms. The most abundant molecules are C ₆₀ , then C ₇₀ and C ₈₄ , followed by C ₇₆ , C ₇₈ , C _{82 ,} etc. with relatively small contents. In addition, since the inside of the carbon cage of fullerenes is a cavity structure, the internal cavity can embed different atoms, ions or clusters of atoms, which is called embedded fullerene, such as La@C ₆₀ , indicating La embedded. In the cage structure of C ₆₀ , @ denotes at, the image expresses the meaning of embedded.

公开于该背景技术部分的信息仅仅旨在增加对本发明的总体背景的理解，而不应当被视为承认或以任何形式暗示该信息构成已为本领域一般技术人员所公知的现有技术。The information disclosed in this Background section is only intended to provide an understanding of the general background of the invention, and should not be construed as an admission

发明内容Summary of the invention

本发明的目的在于提供一种富勒烯结构在制备治疗糖尿病及其并发症的药物中的应用。本发明的另一目的在于提供一种使用上述富勒烯结构治疗糖尿病及其并发症的药物组合物及方法。本发明中涉及的治疗糖尿病及其并发症的有效成分富勒烯结构能在相对较短的时间内有效的降低血糖，显著增加糖耐量，增加胰岛素的敏感性，降低胰岛素抵抗，从而从根本上治疗糖尿病引起的肝肾等功能损伤、血液粘度增加、伤口难以愈合、心脑血管病等并发症。It is an object of the present invention to provide a use of a fullerene structure for the preparation of a medicament for the treatment of diabetes and its complications. Another object of the present invention is to provide a pharmaceutical composition and method for treating diabetes and its complications using the above fullerene structure. The fullerene structure of the active ingredient for treating diabetes and its complications involved in the present invention can effectively lower blood sugar in a relatively short period of time, significantly increase glucose tolerance, increase insulin sensitivity, and reduce insulin resistance, thereby fundamentally Treatment of diabetes-induced liver and kidney damage, increased blood viscosity, difficult to heal wounds, cardiovascular and cerebrovascular diseases and other complications.

为了实现目的，本发明提供了以下技术方案：In order to achieve the object, the present invention provides the following technical solutions:

一种富勒烯结构在制备治疗糖尿病及其并发症的药物中的应用，其中所述富勒烯结构包括至少一种选自下组的有效成分：油溶性的富勒烯、油溶性的内嵌金属富勒烯、所述油溶性的富勒烯和所述油溶性的内嵌金属富勒烯的组合物、水溶性的富勒烯、水溶性的内嵌金属富勒烯、所述水溶性的富勒烯和所述水溶性的内嵌金属富勒烯的组合物、以上六者的可药用的酯或以上六者的可药用的盐。A use of a fullerene structure for the preparation of a medicament for treating diabetes and a complication thereof, wherein the fullerene structure comprises at least one active ingredient selected from the group consisting of oil-soluble fullerenes, oil-soluble internals a metal-filled fullerene, a composition of the oil-soluble fullerene and the oil-soluble inlaid metal fullerene, a water-soluble fullerene, a water-soluble inlaid metal fullerene, the water-soluble A composition of a sexual fullerene and the water-soluble inlaid metal fullerene, a pharmaceutically acceptable ester of the above six or a pharmaceutically acceptable salt of the above six.

本发明还提供了一种治疗糖尿病及其并发症的方法，包括向需要治疗糖尿病及其并发症的受试者施用有效量的富勒烯结构，所述富勒烯结构包括至少一种选自下组的有效成分：油溶性的富勒烯、油溶性的内嵌金属富勒烯、所述油溶性的富勒烯和所述油溶性的内嵌金属富勒烯的组合物、水溶性的富勒烯、水溶性的内嵌金属富勒烯、所述水溶性的富勒烯和所述水溶性的内嵌金属富勒烯的组合物、以上六者的可药用的酯或以上六者的可药用的盐。The invention also provides a method of treating diabetes and its complications comprising administering to a subject in need of treatment for diabetes and its complications an effective amount of a fullerene structure comprising at least one selected from the group consisting of Active ingredients of the lower group: oil-soluble fullerenes, oil-soluble inlaid metal fullerenes, oil-soluble fullerenes and oil-soluble inlaid metal fullerenes, water-soluble a fullerene, a water-soluble inlaid metal fullerene, a water-soluble fullerene and the water-soluble inlaid metal fullerene composition, a pharmaceutically acceptable ester of the above six or above A pharmaceutically acceptable salt.

本发明还提供了一种治疗糖尿病及其并发症的药物组合物，包括至少一种选自下组的富勒烯结构：油溶性的富勒烯、油溶性的内嵌金属富勒烯、所述油溶性的富勒烯和所述油溶性的内嵌金属富勒烯的组合物、水溶性的富勒烯、水溶性的内嵌金属富勒烯、所述水溶性的富勒烯和所述水溶性的内嵌金属富勒烯的组合物、以上六者的可药用的酯、以上六者的可药用的盐；所述药物组合物还包括可药用的载体、可药用的稀释剂和可药用的赋形剂中的至少一种。The present invention also provides a pharmaceutical composition for treating diabetes and its complications, comprising at least one fullerene structure selected from the group consisting of oil-soluble fullerenes, oil-soluble inlaid metal fullerenes, and a composition of oil-soluble fullerenes and the oil-soluble inlaid metal fullerenes, water-soluble fullerene a olefin, a water-soluble inlaid metal fullerene, a water-soluble fullerene, and a water-soluble inlaid metal fullerene composition, a pharmaceutically acceptable ester of the above six, and the above six A pharmaceutically acceptable salt; the pharmaceutical composition further comprising at least one of a pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent, and a pharmaceutically acceptable excipient.

上述应用、方法或药物组合物在另一种实施方式中，所述油溶性的富勒烯包括碳笼外表面包覆有油溶液的富勒烯。In another embodiment, the oil-soluble fullerene comprises a fullerene having an outer surface of the carbon cage coated with an oil solution.

上述应用、方法或药物组合物在另一种实施方式中，所述油溶性的内嵌金属富勒烯包括碳笼外表面包覆有油溶液的内嵌金属富勒烯。In another embodiment, the oil-soluble inlaid metal fullerene comprises an inlaid metal fullerene having an outer surface of the carbon cage coated with an oil solution.

上述应用、方法或药物组合物在另一种实施方式中，所述油溶液可以为单一组分的油，也可以为不同油溶液形成的混合油。通常为植物油，如橄榄油，亚麻籽油，葵花籽油，玉米胚油，大豆油等，也包括动物油脂，如角鲨烷等。The above application, method or pharmaceutical composition In another embodiment, the oil solution may be a single component oil or a mixed oil formed of different oil solutions. Usually vegetable oils, such as olive oil, linseed oil, sunflower oil, corn germ oil, soybean oil, etc., also include animal fats such as squalane.

上述应用、方法或药物组合物在另一种实施方式中，所述碳笼外表面包覆有油溶液的富勒烯是通过将原料富勒烯进行油溶性改性获得的，所述碳笼外表面包覆有油溶液的内嵌金属富勒烯是通过将原料内嵌金属富勒烯进行油溶性改性获得的。In another embodiment, the fullerene coated with an oil solution on the outer surface of the carbon cage is obtained by oil-soluble modification of the raw material fullerenes, the carbon cage The inlaid metal fullerene having an outer surface coated with an oil solution is obtained by oil-soluble modification of a metal fullerene embedded in a raw material.

上述应用、方法或药物组合物在另一种实施方式中，所述油溶性改性为将原料富勒烯和原料内嵌金属富勒烯中的至少一种分散于所述油溶液中，得到油溶性改性液；具体分散的手段可以是将原料与油溶液的混合液经球磨或超声后，依次经离心去除沉淀，然后所得上层液过滤，即得。In another embodiment, the oil-soluble modification is to disperse at least one of a raw material fullerene and a raw material inlaid metal fullerene in the oil solution to obtain The oil-soluble modified liquid; the specific dispersion means that the mixture of the raw material and the oil solution may be subjected to ball milling or ultrasonication, and then the precipitate is removed by centrifugation, and then the supernatant liquid is obtained by filtration.

上述应用、方法或药物组合物在另一种实施方式中，油溶性改性液中有效成分的浓度为0.01-100mg/mL，该范围的公开应当被视为是范围内所有数值的公开，可选的有0.01-10mg/mL，10-20mg/mL，20-30mg/mL，30-40mg/mL等。In another embodiment, the concentration of the active ingredient in the oil-soluble modified liquid is 0.01-100 mg/mL, and the disclosure of the range should be regarded as the disclosure of all the values in the range. The selected ones are 0.01-10 mg/mL, 10-20 mg/mL, 20-30 mg/mL, 30-40 mg/mL, and the like.

上述应用、方法或药物组合物在另一种实施方式中，所述油溶性改性的过程中，每1ml油溶液中分散0.05-1000mg富勒烯原料和/或内嵌金属富勒烯原料，该范围的公开应当被视为是范围内所有数值的公开，可选的有0.05-1mg，0.05-10mg，0.05-100mg等。In another embodiment, in the oil-soluble modification process, 0.05-1000 mg of fullerene raw material and/or embedded metal fullerene raw material are dispersed per 1 ml of the oil solution. The disclosure of this range should be considered as a disclosure of all values in the range, optionally 0.05-1 mg, 0.05-10 mg, 0.05-100 mg, and the like.

上述应用、方法或药物组合物在另一种实施方式中，将所述混合液经球磨或超声30min-15h。 In another embodiment, the mixture is subjected to ball milling or sonication for 30 min to 15 h.

上述应用、方法或药物组合物在另一种实施方式中，混合液经球磨或超声后，离心前，还包括将所述混合液置于阴凉干燥避光保存，静置一定的时间的步骤。可选的，一定的时间指2h-24h。In another embodiment, the above-mentioned application, method or pharmaceutical composition, after ball milling or ultrasonication, before centrifugation, further comprises the step of allowing the mixture to be stored in a cool, dry, dark, and allowed to stand for a certain period of time. Optional, a certain time refers to 2h-24h.

上述应用、方法或药物组合物在另一种实施方式中，所述水溶性的富勒烯包括选自下组中的一种或多种：(1)碳笼外表面修饰有亲水基团的富勒烯；(2)碳笼外表面被亲水性生物小分子包裹的富勒烯；(3)被具有生物相容性的载体材料负载的富勒烯；(4)自组装形成的水溶性超分子体系富勒烯。In another embodiment, the water-soluble fullerene comprises one or more selected from the group consisting of: (1) the outer surface of the carbon cage is modified with a hydrophilic group. Fullerene; (2) fullerenes surrounded by hydrophilic biomolecules on the outer surface of the carbon cage; (3) fullerenes supported by a biocompatible carrier material; (4) formed by self-assembly Water-soluble supramolecular system fullerene.

上述应用、方法或药物组合物在另一种实施方式中，所述水溶性的内嵌金属富勒烯包括选自下组中的一种或多种：(1)碳笼外表面修饰有亲水基团的内嵌金属富勒烯；(2)碳笼外表面被亲水性生物小分子包裹的内嵌金属富勒烯；(3)被具有生物相容性的载体材料负载的内嵌金属富勒烯；(4)自组装形成的水溶性超分子体系内嵌金属富勒烯。In another embodiment, the water-soluble inlaid metal fullerene comprises one or more selected from the group consisting of: (1) the outer surface of the carbon cage is modified with a pro Water-incorporated metal fullerenes; (2) inlaid metal fullerenes surrounded by hydrophilic biomolecules on the outer surface of the carbon cage; (3) inlaid by a biocompatible carrier material Metal fullerenes; (4) Self-assembled metal-filled fullerene formed by self-assembly of a water-soluble supramolecular system.

上述应用、方法或药物组合物在另一种实施方式中，所述亲水基团包括羟基、羧基、巯基和氨基中的一种或多种。In another embodiment, the hydrophilic group includes one or more of a hydroxyl group, a carboxyl group, a thiol group, and an amino group.

上述应用、方法或药物组合物在另一种实施方式中，所述水溶性的内嵌金属富勒烯包括水溶性羟基化Gd@C₈₂；所述水溶性的富勒烯包括水溶性羟基化C₆₀或水溶性羟基化C₇₀。In another embodiment, the water-soluble inlaid metal fullerene comprises water-soluble hydroxylated Gd@C ₈₂ ; the water-soluble fullerene comprises water-soluble hydroxylation C ₆₀ or water-soluble hydroxylated C ₇₀ .

上述应用、方法或药物组合物在另一种实施方式中，所述亲水性生物小分子包括氨基酸和肽链中的至少一种。In another embodiment, the hydrophilic bio-small molecule comprises at least one of an amino acid and a peptide chain.

上述应用、方法或药物组合物在另一种实施方式中，所述具有生物相容性的载体材料包括脂质体和细胞膜载体的至少一种。In another embodiment of the above application, method or pharmaceutical composition, the biocompatible carrier material comprises at least one of a liposome and a cell membrane carrier.

上述应用、方法或药物组合物在另一种实施方式中，所述具有生物相容性的载体材料为医学中常用的药物载体，包括脂质体、细胞膜载体中的至少一种。可选的，所述聚合物胶束为聚乙丙交酯聚乙二醇(PEG-PLGA)、聚赖氨酸或壳聚糖；所述蛋白质为白蛋白或转铁蛋白。In another embodiment, the biocompatible carrier material is a pharmaceutical carrier commonly used in medicine, including at least one of a liposome and a cell membrane carrier. Optionally, the polymer micelle is polyglycolide polyethylene glycol (PEG-PLGA), polylysine or chitosan; the protein is albumin or transferrin.

上述应用、方法或药物组合物在另一种实施方式中，所述水溶性的富勒烯是通过对原料富勒烯进行水溶性改性获得的；所述水溶性的内嵌金属富勒烯是通过对原料内嵌金属富勒烯进行水溶性改性获得的。In another embodiment, the water-soluble fullerene is obtained by water-soluble modification of a raw material fullerene; the water-soluble inlaid metal fullerene It is obtained by water-soluble modification of the metal fullerene embedded in the raw material.

上述应用、方法或药物组合物在另一种实施方式中，所述水溶性改性的方法为以下方法中的任一种：(1)表面修饰亲水基团的方法一般在碱的作用下通过固液或者液液反应实现，具体为将原料富勒烯和原料内嵌金属富勒烯中的至少一种与双氧水和碱(碱具体可为氢氧化钠或者氢氧化钾)混合并进行反应，再用乙醇洗涤，然后透析，即可得到与原料相应的水溶性羟基衍生物。如果需要获得水溶性氨基化衍生物，将上述步骤中的氢氧化物替换成氨水即可。(2)物理包覆的方法可以将原料富勒烯和原料内嵌金属富勒烯中的至少一种与聚乙二醇、聚乙烯吡咯烷酮和环糊精中的至少一种混合并进行球磨或超声等就可以得到与原料相应的被包覆的水溶性富勒烯结构，如聚乙二醇包覆的富勒烯和/或聚乙二醇包覆的内嵌金属富勒烯，聚乙烯吡咯烷酮包覆的富勒烯和/或聚乙烯吡咯烷酮包覆的内嵌金属富勒烯。In another embodiment, the method of water-soluble modification is any one of the following methods: (1) a method of surface modifying a hydrophilic group generally in the action of a base The solution is carried out by a solid-liquid or liquid-liquid reaction, in particular, mixing at least one of the raw material fullerenes and the raw material embedded metal fullerenes with hydrogen peroxide and a base (the alkali may specifically be sodium hydroxide or potassium hydroxide) The reaction is further washed with ethanol and then dialyzed to obtain a water-soluble hydroxy derivative corresponding to the starting material. If it is desired to obtain a water-soluble aminated derivative, the hydroxide in the above step may be replaced with aqueous ammonia. (2) Physical coating method at least one of raw material fullerenes and raw material embedded metal fullerenes may be mixed with at least one of polyethylene glycol, polyvinyl pyrrolidone and cyclodextrin and ball-milled or Ultrasonic or the like can obtain a coated water-soluble fullerene structure corresponding to the raw material, such as polyethylene glycol-coated fullerene and/or polyethylene glycol-coated inlaid metal fullerene, polyethylene Pyrrolidone-coated fullerene and/or polyvinylpyrrolidone-coated inlaid metal fullerenes.

上述应用、方法或药物组合物在另一种实施方式中，称取50～300mg的C₆₀或C₇₀或Gd@C₈₂固体，5～30ml 20～40％的双氧水，2～20ml 1M-3M的碱溶液，在50～100℃的条件下混合，至相应C₆₀或C₇₀或Gd@C₈₂固体全部溶解。在此描述中，表现的是各物质之间的比例关系，实际应用中并不受50～300mg、5～30ml和2～20ml具体反应规模的限制，可按照比例进行扩大。In another embodiment, the above application, method or pharmaceutical composition is weighed 50 to 300 mg of C ₆₀ or C ₇₀ or Gd@C ₈₂ solid, 5 to 30 ml of 20 to 40% hydrogen peroxide, and 2 to 20 ml of 1 M to 3 M. The alkali solution is mixed at 50 to 100 ° C until the corresponding C ₆₀ or C ₇₀ or Gd@C ₈₂ solid is completely dissolved. In this description, the proportional relationship between the substances is expressed. In practice, it is not limited by the specific reaction scale of 50-300 mg, 5-30 ml and 2-20 ml, and can be expanded according to the ratio.

上述应用、方法或药物组合物在另一种实施方式中，所述原料富勒烯包括一种或多种通式为C_2m的由碳原子组成的笼状结构，30≤m≤60，例如；C₆₀，C₇₀，C₈₄等。In another embodiment, the raw material fullerene comprises one or more cage structures consisting of carbon atoms of the formula C _2m , 30 ≤ m ≤ 60, for example ; C ₆₀ , C ₇₀ , C _{84 ,} etc.

上述应用、方法或药物组合物在另一种实施方式中，所述内嵌金属富勒烯原料包括M@C_2n、M₂@C_2n、MA@C_2n、M₃N@C_2n、M₂C₂@C_2n、M₂S@C_2n、M₂O@C_2n和M_xA_3-xN@C_2n中的一种或多种，其中：M、A均代表金属元素且M、A均选自镧系金属元素、Sc、和Y中的任意一种，30≤n≤60；0≤x≤3。N代表氮元素，C代表碳元素，S代表硫元素，镧系金属元素包括La、Ce、Pr、Nd、Pm、Sm、Eu、Gd、Tb、Dy、Ho、Er、Tm、Yb和Lu。例如：Gd@C₈₂。In another embodiment, the inlaid metal fullerene raw material comprises M@C _2n , M ₂ @C _2n , MA@C _2n , M ₃ N@C _2n , M _{One or more of 2} C ₂ @C _2n , M ₂ S@C _2n , M ₂ O@C _2n and M _x A _3-x N@C _2n , wherein: M and A both represent metal elements and M And A is selected from any one of a lanthanide metal element, Sc, and Y, 30 ≤ n ≤ 60; 0 ≤ x ≤ 3. N represents a nitrogen element, C represents a carbon element, S represents a sulfur element, and lanthanide metal elements include La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu. For example: Gd@C ₈₂ .

上述应用、方法或药物组合物在另一种实施方式中，所述糖尿病为一型糖尿病或二型糖尿病。In another embodiment of the above application, method or pharmaceutical composition, the diabetes is type 1 diabetes or type 2 diabetes.

上述应用、方法或药物组合物在另一种实施方式中，所述糖尿病并发症为一型糖尿病引起的并发症或二型糖尿病引起的并发症。In another embodiment of the above application, method or pharmaceutical composition, the diabetic complication is a complication caused by type 1 diabetes or a complication caused by type 2 diabetes.

上述应用、方法或药物组合物在另一种实施方式中，所述糖尿病并发症包括糖尿病性心脑血管病，可选的为微血管和/或大血管病变引起的心脑血管疾病，如：动脉粥样硬化；糖尿病肾病；糖尿病皮肤病，如：糖尿病溃疡，伤口愈合困难；糖尿病引起的高粘稠血症；糖尿病神经病变，如：中风；糖尿病眼部并发症，如：视网膜病变；糖尿病病足。In another embodiment, the diabetic complication includes diabetic cardio-cerebral vascular disease, optionally cardiovascular and cerebrovascular diseases caused by microvascular and/or macrovascular diseases, such as: arteries Atherosclerosis; diabetic nephropathy; diabetic skin diseases such as: diabetic ulcers, Difficulty in wound healing; hyperviscosity caused by diabetes; diabetic neuropathy, such as: stroke; diabetic eye complications, such as: retinopathy; diabetic foot.

上述应用、方法或药物组合物在另一种实施方式中，所述治疗糖尿病并发症包括：1)使糖尿病引起的血糖升高趋于正常；2)使胰岛的大小和胰岛细胞的数量趋于正常；3)调节胰岛素分泌，减轻胰岛素抵抗，增加胰岛素敏感性，使糖耐量和胰岛素耐量趋于正常；4)治疗糖尿病引起的冠心病；5)使糖尿病肾病引起的肝肾指标失常趋于正常(肝肾指标包括：谷丙转氨酶，谷草转氨酶，血肌酐，尿素氮)，使尿蛋白明显降低；6)使伤口愈合加快；7)降低糖尿病引起的血液粘度增加；8)降低血脂(血脂指标包括：总胆固醇，甘油三酯，高低密度脂)；9)治疗糖尿病病足；10)治疗糖尿病性视网膜病变、与糖尿病相关的葡萄膜炎和糖尿病性白内障；11)使糖化血红蛋白显著降低；12)使糖基化产物(AGE)降低；13)调节机体免疫及炎症；14)改善血液流变异常，抑制血小板高聚集性；15)预防及改善神经功能不全。In another embodiment, the treating diabetes complications include: 1) making the blood glucose elevation caused by diabetes tend to normal; 2) concentrating the size of the islets and the number of islet cells Normal; 3) regulate insulin secretion, reduce insulin resistance, increase insulin sensitivity, make glucose tolerance and insulin tolerance tend to normal; 4) treat coronary heart disease caused by diabetes; 5) make liver and kidney index abnormalities caused by diabetic nephropathy tend to normal (Hepatic and kidney indicators include: alanine aminotransferase, aspartate aminotransferase, serum creatinine, urea nitrogen), which significantly reduces urine protein; 6) accelerate wound healing; 7) reduce blood viscosity increase caused by diabetes; 8) lower blood lipids (blood lipid index) Including: total cholesterol, triglycerides, high and low density lipids; 9) treatment of diabetic foot; 10) treatment of diabetic retinopathy, diabetes-related uveitis and diabetic cataract; 11) significantly reduced glycated hemoglobin; 12) reduce the glycosylation product (AGE); 13) regulate the body's immunity and inflammation; 14) improve blood rheology abnormalities, inhibit platelet aggregation; 15) Prevention and improvement of neurological dysfunction.

上述应用中的药物或上述药物组合物在另一种实施方式中，该药物或药物组合物可以是片剂、丸剂、散剂、锭剂、小药囊、扁囊剂、酏剂、悬浮剂、乳剂、溶液剂、糖浆剂、气溶胶、软膏、软和硬明胶胶囊、栓剂、无菌注射溶液或无菌包装粉针剂的制剂。本发明中将有效成分制备成药物或药物组合物的方法可采用本领域普通技术人员公知的方法来制备，使其在施用于受试者后速释、缓释或延迟释放有效成分，例如：有效成分可以与载体混合，用载体稀释或者包封在载体中。In another embodiment, the drug or the pharmaceutical composition may be a tablet, a pill, a powder, a lozenge, a sachet, a cachet, an elixir, a suspending agent, Formulations of emulsions, solutions, syrups, aerosols, ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions or sterile packaging powders. The method of preparing an active ingredient into a pharmaceutical or pharmaceutical composition in the present invention can be prepared by a method known to those skilled in the art to provide an immediate release, sustained release or delayed release of the active ingredient after administration to a subject, for example: The active ingredient can be mixed with the carrier, diluted with the carrier or enclosed in a carrier.

上述应用中的药物或上述药物组合物在另一种实施方式中，适宜作为载体、赋形剂和稀释剂的一些实例包括乳糖、右旋糖、蔗糖、山梨醇、甘露醇、淀粉、树脂、***胶、磷酸钙、海藻酸盐、西黄蓍胶、明胶、硅酸钙、微晶纤维素、聚乙烯吡咯烷酮、纤维素、水糖浆(water syrup)、甲基纤维素、尼泊金甲酯和丙酯、滑石粉、硬脂酸镁和液状石蜡。The drug or the above pharmaceutical composition in the above application. In another embodiment, some examples suitable as carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starch, resins, Acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, methylcellulose, methylparaben And propyl ester, talc, magnesium stearate and liquid paraffin.

上述应用中的药物或上述药物组合物在另一种实施方式中，该药物或药物组合物还可以另外包括润滑剂、润湿剂、乳化和悬浮剂、防腐剂、甜味剂或矫味剂等助剂。The medicament or the above pharmaceutical composition in the above application. In another embodiment, the medicament or pharmaceutical composition may additionally comprise a lubricant, a wetting agent, an emulsifying and suspending agent, a preservative, a sweetener or a flavoring agent. And other additives.

上述方法在另一种实施方式中，所述受试者为人或动物，动物可以为哺乳动物，如小鼠、豚鼠、大鼠、狗、兔子、猴子等。 In another embodiment, the subject is a human or an animal, and the animal can be a mammal such as a mouse, a guinea pig, a rat, a dog, a rabbit, a monkey, or the like.

上述方法在另一种实施方式中，所述有效成分的施用剂量为1mg/kg/d-1000mg/kg/d，可选的为1-100mg/kg/d，10mg/kg/d-100mg/kg/d，1-20mg/kg/d，1-10mg/kg/d，施用疗程可以为5天-30天，根据病情可短期服用或长期服用；有效成分的施用方式可以为口服、注射(如：静脉注射)或腹腔给药。注射后有效成分进入体内直接经血液循环发挥作用，无需渗透，所用的药剂量小，疗效高；口服摄入，经消化***过滤吸收，副作用更小，疗效显著。In another embodiment, the active ingredient is administered at a dose of from 1 mg/kg/d to 1000 mg/kg/d, optionally from 1 to 100 mg/kg/d, and from 10 mg/kg/d to 100 mg/ Kg / d, 1-20mg / kg / d, 1-10mg / kg / d, the course of application can be 5 days - 30 days, depending on the condition can be taken short-term or long-term use; the active ingredient can be administered orally, injection ( Such as: intravenous injection or intraperitoneal administration. After the injection, the active ingredient enters the body and directly acts through the blood circulation without permeation. The amount of the medicament used is small and the curative effect is high; the oral intake is filtered and absorbed by the digestive system, and the side effects are smaller and the curative effect is remarkable.

上述应用中的药物或上述药物组合物在另一种实施方式中，当所述药物或所述药物组合物以液体形式存在时，有效成分在所述药物或所述药物组合物中的浓度为0.01-100mg/mL，可选的为0.01-10mg/mL，0.01-20mg/mL，0.01-30mg/mL，0.01-40mg/mL；当所述药物或所述药物组合物以固体形式存在时，有效成分在所述药物或所述药物组合物中的浓度为0.01-50mg/g，可选的为0.01-10mg/g，0.01-20mg/g，0.01-30mg/g，0.01-40mg/g。The drug or the above pharmaceutical composition in the above application. In another embodiment, when the drug or the pharmaceutical composition is present in a liquid form, the concentration of the active ingredient in the drug or the pharmaceutical composition is 0.01-100 mg/mL, optionally 0.01-10 mg/mL, 0.01-20 mg/mL, 0.01-30 mg/mL, 0.01-40 mg/mL; when the drug or the pharmaceutical composition is present in a solid form, The concentration of the active ingredient in the drug or the pharmaceutical composition is from 0.01 to 50 mg/g, alternatively from 0.01 to 10 mg/g, from 0.01 to 20 mg/g, from 0.01 to 30 mg/g, from 0.01 to 40 mg/g.

上述应用中的药物或上述药物组合物在另一种实施方式中，所述水溶性的富勒烯和/或水溶性的内嵌金属富勒烯在制剂中的浓度为0.01-100mg/mL；所述油溶性的富勒烯和/或油溶性的内嵌金属富勒烯在制剂中的浓度为500ppm-10000ppm(mg/kg)。In another embodiment, the water-soluble fullerene and/or water-soluble inlaid metal fullerene is in the formulation at a concentration of 0.01-100 mg/mL; The oil-soluble fullerene and/or oil-soluble inlaid metal fullerene is present in the formulation at a concentration of from 500 ppm to 10,000 ppm (mg/kg).

本发明所用的术语“治疗”包括其通常被接受的含义，该含义包括阻止、预防、抑制、改善以及减缓、停止或逆转所产生症状或预期病变的发展。照此，本发明涵盖治疗性和预防性的施用。As used herein, the term "treatment" includes its generally accepted meaning, which includes preventing, preventing, inhibiting, ameliorating, and slowing, halting, or reversing the development of a symptom or a desired condition. As such, the invention encompasses both therapeutic and prophylactic administration.

本发明所用的术语“有效成分”、“有效成分富勒烯结构”或“富勒烯结构”均指的是油溶性的富勒烯、油溶性的内嵌金属富勒烯、所述油溶性的富勒烯和所述油溶性的内嵌金属富勒烯的组合物、水溶性的富勒烯、水溶性的内嵌金属富勒烯、所述水溶性的富勒烯和所述水溶性的内嵌金属富勒烯的组合物、以上六者的可药用的酯或以上六者的可药用的盐中的至少一种。The term "active ingredient", "active ingredient fullerene structure" or "fullerene structure" as used herein, refers to oil-soluble fullerenes, oil-soluble inlaid metal fullerenes, and oil solubility. a composition of fullerene and the oil-soluble inlaid metal fullerene, a water-soluble fullerene, a water-soluble inlaid metal fullerene, the water-soluble fullerene, and the water-soluble The composition of the metal fullerene embedded, the pharmaceutically acceptable ester of the above six or at least one of the pharmaceutically acceptable salts of the above six.

本发明所用的术语“有效量”指有效成分经单次或多次施用于患者而给所诊断或所治疗的患者提供预期效应的量或剂量。有效量可由所参与的诊断医师作为本领域技术人员通过已知技术以及在类似情形下所得的观察结果而确定。在确定所施用有效成分的有效量或剂量时，所参与的诊断医师应考虑多种因素，所述因素包括但不限于：哺乳动物的种属；体积、年龄及一般健康；所涉及的具体疾病；该疾病的涉入程度或严重程度；个体患者的响应；所施用的具体化合物；给药模式；所施用制剂的生物利用度性质；所选择的给药方案；伴随药物疗法的使用；以及其它相关的情形。The term "effective amount" as used herein, refers to an amount or dose of the active ingredient that is administered to a patient in a single or multiple administrations to provide a desired effect to the patient being diagnosed or treated. The effective amount can be determined by the participating diagnostician as a result of known techniques by those skilled in the art and in similar circumstances. In determining the effective amount or dose of the active ingredient to be administered, the participating diagnostician should consider a variety of factors including, but not limited to, the mammalian species; volume, age, and general health. The specific disease involved; the extent or severity of the disease; the response of the individual patient; the specific compound administered; the mode of administration; the bioavailability properties of the administered formulation; the chosen dosing regimen; Use of drug therapy; and other related situations.

本发明所用的术语“原料富勒烯”是指没有经过水溶性改性或油溶性改性的富勒烯，即富勒烯本体。The term "raw material fullerene" as used in the present invention means a fullerene which is not subjected to water-soluble modification or oil-soluble modification, that is, a fullerene body.

本发明所用的术语“原料内嵌金属富勒烯”是指没有经过水溶性改性或油溶性改性的内嵌金属富勒烯，即内嵌金属富勒烯本体。The term "inlaid metal fullerene in the raw material" as used in the present invention means an inlaid metal fullerene which is not subjected to water-soluble modification or oil-soluble modification, that is, an inlaid metal fullerene body.

为了方便计量，本发明中所有关于水溶性的富勒烯、水溶性的金属富勒烯、油溶性的富勒烯或油溶性的金属富勒烯的具体含量、浓度等定量的限定均是以其对应的富勒烯本体或内嵌金属富勒烯本体的具体含量、浓度等来衡量的，例如：水溶性的富勒烯在制剂中的浓度为0.01-100mg/mL是指可检测到水溶性的富勒烯中的富勒烯本体碳笼在制剂中的浓度为0.01-100mg/mL；又例如：碳笼外表面包覆有油溶液的富勒烯的含量为100μM是指油溶液内的富勒烯本体碳笼的含量是100μM。其中：金属富勒烯和内嵌金属富勒烯可以通过电感耦合等离子体发射光谱仪(ICP)进行定量测定。In order to facilitate the metering, all the specific contents, concentrations, etc. of the water-soluble fullerene, the water-soluble metal fullerene, the oil-soluble fullerene or the oil-soluble metal fullerene in the present invention are quantitatively limited. The specific content and concentration of the corresponding fullerene body or the embedded metal fullerene body, for example, the concentration of the water-soluble fullerene in the preparation is 0.01-100 mg/mL, which means that water solubility can be detected. The concentration of the fullerene bulk carbon cage in the fullerene is 0.01-100 mg/mL in the preparation; for example, the content of fullerene coated with the oil solution on the outer surface of the carbon cage is 100 μM means that the oil solution is The content of the fullerene bulk carbon cage was 100 μM. Among them: metal fullerenes and embedded metal fullerenes can be quantitatively determined by inductively coupled plasma optical emission spectrometry (ICP).

与现有技术相比，本发明的有益效果为：Compared with the prior art, the beneficial effects of the present invention are:

1、本发明中有效成分油溶性富勒烯结构或水溶性富勒烯结构均保持了完整的碳笼结构，油溶性富勒烯结构具有脂溶性特性，其进入机体内通过消化吸收进入血液循环或通过直接的器官渗透进入各个器官中；水溶性富勒烯结构在生物体内随着血液循环输送至各个器官中发挥作用。有效成分进入胰腺，改善胰腺以及胰腺中胰岛的微环境，修正胰岛的结构，减轻胰腺的功能性损伤，减小胰岛β细胞的损伤，利于胰腺正常分泌胰岛素，并能减轻胰岛素抵抗，从而达到降低血糖的作用；另外有效成分进入肝、肾等器官，减少糖、蛋白质以及脂质的氧化反应，减少糖基化终产物，从而减轻各种相应的糖尿病并发症。1. In the present invention, the oil-soluble fullerene structure or the water-soluble fullerene structure of the active ingredient maintains a complete carbon cage structure, and the oil-soluble fullerene structure has fat-soluble characteristics, which enter the body and pass through digestion and absorption into the blood circulation. Or through direct organ penetration into various organs; water-soluble fullerene structure plays a role in the body to the various organs with blood circulation. The active ingredient enters the pancreas, improves the microenvironment of pancreatic islets in the pancreas and pancreas, corrects the structure of the islets, reduces the functional damage of the pancreas, reduces the damage of islet β cells, facilitates the normal secretion of insulin from the pancreas, and reduces insulin resistance, thereby reducing The role of blood sugar; the active ingredient enters the liver, kidney and other organs, reducing the oxidation of sugar, protein and lipids, reducing the glycation end products, thereby alleviating various complications of diabetes.

2、有效成分油溶性富勒烯结构或水溶性富勒烯结构均具有良好的清除自由基的效果并能改善机体的氧化还原水平，而且有效成分可以被快速代谢，对体内器官不会有毒性，具有良好的生物相容性。2, the active ingredient oil-soluble fullerene structure or water-soluble fullerene structure has a good effect of scavenging free radicals and can improve the body's redox level, and the active ingredients can be quickly metabolized, no toxicity to internal organs , has good biocompatibility.

3、现有技术中糖尿病的治疗都需要药物长期作用，而本发明中有效成分富勒烯结构能在相对较短的时间内有效的降低血糖，经过服用5-30天后，血糖值可显著降低，糖耐量可显著增加，从而从根本上治疗糖尿病及其引起的肝肾损伤、心血管病、伤口愈合缓慢等并发症。本发明中油溶性富勒烯结构可以配合降血糖药物使用，降血糖的同时能够减轻副作用，改善并发症状况。3. The treatment of diabetes in the prior art requires long-term effects of the drug, and the active ingredient in the present invention The fullerene structure can effectively lower blood sugar in a relatively short period of time. After taking 5-30 days, the blood sugar level can be significantly reduced, and the glucose tolerance can be significantly increased, thereby fundamentally treating diabetes and its caused liver and kidney damage, Complications such as cardiovascular disease and slow wound healing. In the present invention, the oil-soluble fullerene structure can be used in combination with a hypoglycemic agent, which can reduce side effects and improve complications while lowering blood sugar.

4、本发明中油溶性富勒烯结构所选用的进行包覆的油溶液本身就可以选择健康无副作用的植物油或动物油，本身就有营养的作用，如橄榄油是一种具有天然保健，美容功效的植物油，能够促进骨骼和神经***的发育，能够美容，抗衰老，并且能够预防心血管疾病等。4. The oil solution coated with the oil-soluble fullerene structure of the present invention can select a vegetable oil or animal oil which has no side effects and has a nutritive effect. For example, olive oil is a natural health care and cosmetic effect. Vegetable oil, which promotes the development of bones and nervous system, can be cosmetic, anti-aging, and can prevent cardiovascular diseases.

附图说明DRAWINGS

图1为实施例2中Gd@C₈₂(OH)_n材料水溶液的照片。Figure 1 is a photograph of an aqueous solution of Gd@C ₈₂ (OH) _n material in Example 2.

图2为实施例3中小鼠给药2周后血清中超氧化物歧化酶和过氧化氢酶的变化图。Fig. 2 is a graph showing changes in serum superoxide dismutase and catalase after administration of mice for 2 weeks in Example 3.

图3为实施例3中小鼠给药2周后血清中丙二醛的含量图。Fig. 3 is a graph showing the content of malondialdehyde in serum after administration of mice for 2 weeks in Example 3.

图4为实施例3中小鼠给药2周后肝损伤指标谷丙转氨酶和谷草转氨酶的检测。Fig. 4 is a graph showing the detection of alanine aminotransferase and aspartate aminotransferase in liver injury indexes after administration of mice in Example 3 for 2 weeks.

图5为实施例3中小鼠给药2周后肾损伤指标测试尿素氮和血肌酐的检测。Fig. 5 is a graph showing the detection of urea nitrogen and serum creatinine in the kidney injury index after administration of the mice in Example 3 for 2 weeks.

图6为实施例3中小鼠给药2周后尿蛋白的变化。Figure 6 is a graph showing changes in urinary protein after administration of mice for 2 weeks in Example 3.

图7为实施例4中Gd@C₈₂(OH)_n材料在生物体内24h的代谢分布。Figure 7 is a graph showing the metabolic distribution of Gd@C ₈₂ (OH) _n material in vivo for 24 h in vivo.

图8为实施例5中Gd@C₈₂(OH)n材料的电子自旋核磁共振(ESR)图，其中，平滑线表示空白对照，非平滑线表示加入Gd@C₈₂(OH)n。Figure 8 is an electron spin nuclear magnetic resonance (ESR) image of the Gd@C ₈₂ (OH)n material in Example 5, wherein the smooth line indicates a blank control and the non-smooth line indicates the addition of Gd@C ₈₂ (OH)n.

图9为实施例6中治疗过程中空腹血糖的变化。Figure 9 is a graph showing changes in fasting blood glucose during the treatment in Example 6.

图10为实施例6中治疗两周后的糖耐受曲线。Figure 10 is a graph showing the glucose tolerance curve after two weeks of treatment in Example 6.

图11为实施例6中治疗两周后的糖耐受实验中120分钟内的曲线下面积柱状图。Figure 11 is a bar graph of the area under the curve within 120 minutes in the glucose tolerance test after two weeks of treatment in Example 6.

图12为实施例6中治疗两周后小鼠体内过氧化氢酶的含量变化。Figure 12 is a graph showing changes in the content of catalase in mice after two weeks of treatment in Example 6.

图13为实施例6中治疗两周后小鼠体内丙二醛的含量变化。Figure 13 is a graph showing changes in the content of malondialdehyde in mice after two weeks of treatment in Example 6.

图14为实施例7中胰岛素耐受实验中的血糖变化曲线。Figure 14 is a graph showing changes in blood glucose in the insulin resistance test in Example 7.

图15为实施例7中给药2周后小鼠的甘油三酯和总胆固醇的含量。 Figure 15 is a graph showing the contents of triglyceride and total cholesterol in mice after administration for 2 weeks in Example 7.

图16为实施例7中给药2周后小鼠脂肪细胞中葡萄糖转运蛋白-4的含量。Figure 16 is a graph showing the content of glucose transporter-4 in mouse adipocytes after administration for 2 weeks in Example 7.

具体实施方式detailed description

下面结合附图，对本发明的具体实施方式进行详细描述，但应当理解本发明的保护范围并不受具体实施方式的限制。The specific embodiments of the present invention are described in detail below with reference to the accompanying drawings, but it is understood that the scope of the present invention is not limited by the specific embodiments.

下述实施例中所使用的实验方法如无特殊说明，均为常规方法。下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。以下实施例所用原料Gd@C₈₂固体粉末购买于厦门福纳新材料科技有限公司，分子量1141，纯度为99.1％。以下实施例所用原料C₆₀固体粉末购买于厦门福纳新材料科技有限公司，分子量720，纯度99％。以下实施例所用原料C₇₀固体粉末购买于厦门福纳新材料科技有限公司，分子量840，纯度99％。The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents and the like used in the following examples are commercially available unless otherwise specified. The raw material Gd@C ₈₂ solid powder used in the following examples was purchased from Xiamen Funa New Material Technology Co., Ltd., with a molecular weight of 1141 and a purity of 99.1%. The raw material C ₆₀ solid powder used in the following examples was purchased from Xiamen Funa New Material Technology Co., Ltd., with a molecular weight of 720 and a purity of 99%. The raw material C ₇₀ solid powder used in the following examples was purchased from Xiamen Funa New Material Technology Co., Ltd., with a molecular weight of 840 and a purity of 99%.

实施例1、油溶性富勒烯或油溶性内嵌金属富勒烯的制备Example 1. Preparation of oil-soluble fullerenes or oil-soluble inlaid metal fullerenes

量取20ml橄榄油，称取20mg C₆₀或20mg C₇₀或20mg Gd@C₈₂，混合搅拌均匀，得到混合液；然后将混合液置于球磨机中球磨10h，球磨结束后将混合液取出，阴凉干燥避光保存，静置4h后离心去除沉淀，然后所得上层液过滤，即得。本申请中将橄榄油与C₆₀按上述方法制备的油溶性富勒烯简称C₆₀-橄榄油，将橄榄油与C₇₀按上述方法制备的油溶性富勒烯简称C₇₀-橄榄油，将橄榄油与Gd@C₈₂按上述方法制备的油溶性富勒烯简称Gd@C₈₂-橄榄油。其中该油溶性改性液中经过改性的原料富勒烯或原料内嵌金属富勒烯的含量为0.8mg/ml。Take 20ml of olive oil, weigh 20mg C ₆₀ or 20mg C ₇₀ or 20mg Gd@C ₈₂ , mix and mix evenly to get a mixture; then put the mixture in a ball mill for 10h ball milling, remove the mixture after ball milling, cool Store in the dark and avoid it. After standing for 4 hours, remove the precipitate by centrifugation, and then filter the obtained supernatant to obtain. The present application will olive oil soluble and C ₆₀ fullerene prepared by the above method referred C ₆₀ - Olive oil, olive oil and oil-soluble C ₇₀ fullerene prepared by the above method referred C ₇₀ - olive oil, the Olive oil and Gd@C ₈₂ oil-soluble fullerenes prepared as described above are abbreviated as Gd@C ₈₂ - olive oil. The content of the modified fullerene or the raw material inlaid metal fullerene in the oil-soluble modified liquid is 0.8 mg/ml.

实施例2、水溶性富勒烯或水溶性内嵌金属富勒烯的制备Example 2. Preparation of water-soluble fullerene or water-soluble inlaid metal fullerene

称取100mg C₆₀或100mg C₇₀或100mg Gd@C₈₂固体，分别加入7ml 30％的双氧水和3ml 10％的氢氧化钠溶液，在50℃的条件下反应至固体全部溶解，然后用乙醇洗涤，反应后使用透析袋透析除去小分子，使用电导率仪监测直至透析完成，超滤离心得到羟基化的水溶性富勒烯或羟基化的水溶性内嵌金属富勒烯。将10％氢氧化钠溶液替换成一定量的30％氨水，可得到氨基化的水溶性富勒烯或氨基化的水溶性内嵌金属富勒烯。Weigh 100mg C ₆₀ or 100mg C ₇₀ or 100mg Gd@C ₈₂ solid, add 7ml 30% hydrogen peroxide and 3ml 10% sodium hydroxide solution, react at 50 ° C until the solid is completely dissolved, then wash with ethanol After the reaction, the small molecule was removed by dialysis using a dialysis bag, and monitoring was performed using a conductivity meter until the completion of dialysis, and the hydroxylated water-soluble fullerene or the hydroxylated water-soluble inlaid metal fullerene was obtained by ultrafiltration centrifugation. An aminated water-soluble fullerene or an aminated water-soluble inlaid metal fullerene can be obtained by replacing a 10% sodium hydroxide solution with a certain amount of 30% aqueous ammonia.

以上，透析后所得的羟基化水溶性空心富勒烯或羟基化水溶性内嵌金属富勒烯中含有较多液体，超滤离心是将其浓缩，但无论是否进行浓缩，原料金属富勒烯或原料内嵌金属富勒烯的水溶性改性已经完成，是否进行浓缩不影响其使用，使用时只要将水溶性富勒烯或水溶性内嵌金属富勒烯调整至合适的浓度即可。Above, the hydroxylated water-soluble hollow fullerene or the hydroxylated water-soluble inlaid metal fullerene obtained after dialysis contains more liquid, and is concentrated by ultrafiltration centrifugation, but whether or not concentrated, raw materials The water-soluble modification of metal fullerenes or metal-filled fullerene in the raw material has been completed, and whether or not the concentration is carried out does not affect the use thereof, and the water-soluble fullerene or the water-soluble inlaid metal fullerene is adjusted to a suitable use. The concentration can be.

图1即为200μM的Gd@C₈₂(OH)_n水溶液照片，可以看出其澄清透明，水溶性好。Fig. 1 is a photograph of a 200 μM aqueous solution of Gd@C ₈₂ (OH) _n , which can be seen to be clear and transparent, and has good water solubility.

实施例3、油溶性富勒烯对糖尿病并发症的治疗Example 3: Treatment of diabetic complications with oil-soluble fullerenes

(1)STZ-高脂诱导的糖尿病小鼠模型的建立(1) Establishment of STZ-high fat-induced diabetes mouse model

本发明采用成熟的高脂喂养STZ(链脲佐菌素)诱导的二型糖尿病模型来研究油溶性富勒烯对糖尿病并发症的治疗作用。The invention adopts a mature high-fat fed STZ (streptozotocin)-induced type 2 diabetes model to study the therapeutic effect of oil-soluble fullerenes on diabetic complications.

购买7-8周的雄性ICR小鼠24只，购自北京大学实验动物中心，随机分为4组，每组6只；其中1组中的6只雄性ICR小鼠作为无糖尿病的健康小鼠对照组(即空白组)，另外18只用于形成糖尿病模型。糖尿病模型的形成方式如下：首先用高脂饲料喂养小鼠4周，然后禁食12h后，快速腹腔注射STZ的柠檬酸溶液，STZ的使用剂量为60mg/kg/d(即按照小鼠体重的每1kg注射60mg的STZ计算)，每天1次，连续注射3天。稳定一周后，检测血糖，空腹血糖>11mmol/L即为成功的2型糖尿病模型，模型在一定时间内显示出了糖尿病肾病、糖尿病肝病、糖尿病血管疾病、伤口愈合缓慢等并发症的症状。Twenty-four male ICR mice purchased from 7-8 weeks were purchased from Peking University Experimental Animal Center and randomly divided into 4 groups, 6 in each group. 6 male ICR mice in 1 group were used as healthy mice without diabetes. The control group (ie, the blank group) and the other 18 were used to form a diabetes model. The diabetes model was formed as follows: First, the mice were fed with high-fat diet for 4 weeks, then fasted for 12 hours, and then quickly injected intraperitoneally with STZ citrate solution. The dose of STZ was 60 mg/kg/d (ie, according to the weight of the mice). Calculated by injecting 60 mg of STZ per 1 kg), once a day for 3 consecutive days. After one week of stable blood glucose test, fasting blood glucose >11mmol/L is a successful type 2 diabetes model. The model shows symptoms of complications such as diabetic nephropathy, diabetic liver disease, diabetic vascular disease, and slow wound healing in a certain period of time.

(2)实验方法(2) Experimental method

实验小鼠分为4组，每组6只，将步骤(1)中6只无糖尿病的健康小鼠对照组(即空白组)作为A组，其给药治疗为施用与D组所用药物等体积的生理盐水；随机取步骤(1)中形成的糖尿病模型小鼠6只作为模型组(简称为B组)，其给药治疗为施用与D组所用药物等体积生理盐水；随机取步骤(1)中形成的糖尿病模型小鼠6只作为橄榄油组(简称为C组)，并施以与D组所用药物等体积的纯橄榄油；随机取步骤(1)中形成的糖尿病模型小鼠6只并施以实施例1制备的C₆₀-橄榄油作为实验组(简称D组)。ABCD组均采用灌胃的方式，每天给药1次，连续2周，D组中C₆₀-橄榄油的给药剂量为20mg/kg/d。The experimental mice were divided into 4 groups, 6 in each group, and 6 healthy mice in the step (1) with no diabetes (ie, the blank group) were used as the group A, and the administration treatment was the administration of the drugs used in the group D and the like. Volume of physiological saline; 6 diabetic model mice formed in step (1) were randomly selected as a model group (abbreviated as group B), and the administration treatment was to apply the same volume of physiological saline as the drug used in group D; 1) Diabetic model mice formed in 6 rats as olive oil group (referred to as group C), and applied with the same volume of pure olive oil as the drug used in group D; randomized diabetic mice formed in step (1) Six of the C ₆₀ - olive oils prepared in Example 1 were applied as an experimental group (referred to as Group D). The ABCD group was administered by intragastric administration once a day for 2 weeks. The dose of C ₆₀ - olive oil in group D was 20 mg/kg/d.

(3)效果测试(3) Effect test

通过上述方法给药2周后测试小鼠体内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和丙二醛(MDA)的含量表征体内的氧化还原水平，通过测试谷丙转氨酶(ALT)，谷草转氨酶(AST)，血肌酐(Cr)和尿素氮(BUN)的含量表征肝肾功能，通过尿蛋白的含量表征糖尿病肾病的程度。Superoxide dismutase (SOD), peroxidation in mice after 2 weeks of administration by the above method Hydrogenase (CAT) and malondialdehyde (MDA) levels characterize redox levels in vivo by testing alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum creatinine (Cr) and urea nitrogen (BUN) levels Liver and kidney function, the extent of diabetic nephropathy is characterized by the amount of urinary protein.

通过实验过程中的观察以及血压结果表明油溶性富勒烯确实对改善糖尿病并发症有好的的作用，具体实验方法和结果如下所示。Observations during the experiment and blood pressure results indicate that oil-soluble fullerenes do have a good effect on improving diabetic complications. The specific experimental methods and results are shown below.

1)体内氧化还原水平测试1) In vivo redox level test

检测指标：SOD和CAT可以体现体内氧化还原的水平；SOD是生物体内重要的抗氧化酶，广泛分布于各种生物体内，其具有特殊的生理活性，是生物体内清除自由基的首要物质；CAT是一种酶类清除剂，它可促使H₂O₂分解为分子氧和水，清除体内的过氧化氢，从而使细胞免于遭受H₂O₂的毒害。MDA为自由基代谢产物，产生更多的自由基时，其含量会增加。Detection indicators: SOD and CAT can reflect the level of redox in the body; SOD is an important antioxidant enzyme in the body, widely distributed in various organisms, it has special physiological activity, is the primary substance in the body to scavenge free radicals; CAT It is an enzyme scavenger that decomposes H ₂ O ₂ into molecular oxygen and water, and removes hydrogen peroxide from the body, thereby protecting cells from H ₂ O ₂ poisoning. MDA is a free radical metabolite, and its content increases when more free radicals are produced.

实验方法：给药两周后的A-D组小鼠通过眼球取血，血液样品室温静置1h后，3500rpm 15min离心后吸取上层血清，利用分光光度法，测定550nm处的吸光度计算得到小鼠体内超氧化物歧化酶(SOD)含量，测定405nm处的吸光度计算得到过氧化氢酶(CAT)的含量，测定532nm处的吸光度计算得到丙二醛(MDA)的含量。以上三种物质的检测均可通过市面可获得的相应试剂盒进行检测。Experimental method: After two weeks of administration, mice in the AD group were bled by eyeballs. The blood samples were allowed to stand at room temperature for 1 h, centrifuged at 3500 rpm for 15 min, and the supernatant serum was aspirated. The absorbance at 550 nm was determined by spectrophotometry. The content of catalase (SOD) was determined by measuring the absorbance at 405 nm, and the content of catalase (MDA) was calculated by measuring the absorbance at 532 nm. The detection of the above three substances can be detected by the corresponding kits available in the market.

实验结果：图2表明患有糖尿病的小鼠体内的超氧化物歧化酶和过氧化氢酶明显低于健康小鼠(参见模型组和空白组的对比)，而经过C₆₀-橄榄油治疗后的小鼠体内超氧化物歧化酶和过氧化氢酶趋于正常水平，如实验组所示。图3表明患有糖尿病的小鼠体内的丙二醛明显高于健康小鼠(参见模型组和空白组的对比)，而经过C₆₀-橄榄油治疗后的小鼠体内丙二醛含量降低，说明合成自由基减少。这三个指标的变化说明了小鼠体内的氧化还原水平得到了调节，相应的氧化应激以及其带来的并发症也会减弱。EXPERIMENTAL RESULTS: Figure 2 shows that superoxide dismutase and catalase are significantly lower in mice with diabetes than in healthy mice (see comparison between model group and blank group), after C ₆₀ - olive oil treatment Superoxide dismutase and catalase in mice tend to normal levels, as shown in the experimental group. Figure 3 shows that malondialdehyde is significantly higher in mice with diabetes than in healthy mice (see comparison between model group and blank group), whereas mice treated with C ₆₀ -olive oil have lower malondialdehyde content. Explain that synthetic free radicals decrease. Changes in these three indicators indicate that the level of redox in the mouse is regulated, and the corresponding oxidative stress and its complications are also reduced.

2)肝肾功能测试2) Liver and kidney function test

检测指标：谷丙转氨酶和谷草转氨酶是表征肝功能的指标，尿素氮和血肌酐是表征肾功能的指标。Detection indicators: alanine aminotransferase and aspartate aminotransferase are indicators of liver function, and urea nitrogen and serum creatinine are indicators of renal function.

实验方法：给药两周后的A-D组小鼠通过眼球取血，血液样品室温静置1h后，3500rpm 15min离心后吸取上层血清，利用自动血生化仪对血清进行测试，检测其中的谷丙转氨酶(ALT)，谷草转氨酶(AST)，血肌酐(Cr) 和尿素氮(BUN)的含量。Experimental method: After two weeks of administration, the mice in the AD group were bled by eyeballs. The blood samples were allowed to stand at room temperature for 1 h, centrifuged at 3500 rpm for 15 min, and the supernatant serum was aspirated. The serum was tested by an automatic blood biochemistry instrument to detect the alanine aminotransferase. (ALT), aspartate aminotransferase (AST), serum creatinine (Cr) And the content of urea nitrogen (BUN).

实验结果：图4表明患有糖尿病的小鼠的谷丙转氨酶和谷草转氨酶的指标远远高于正常小鼠的值(参见模型组和空白组的对比)，但是经过C₆₀-橄榄油治疗后的小鼠谷丙转氨酶和谷草转氨酶的值都趋于正常，而橄榄油组与模型组没有明显差异，这说明油溶性富勒烯能够改善糖尿病引起的肝功能损伤。图5表明患有糖尿病小鼠的尿素氮和血肌酐的指标都明显高于正常小鼠的值(参见模型组和空白组的对比)，但是经过C₆₀-橄榄油治疗后的小鼠尿素氮和血肌酐的值都趋于正常，而橄榄油组与模型组没有明显差异，这说明油溶性富勒烯能够改善糖尿病引起的肾功能损伤。Experimental results: Figure 4 shows that the index of alanine aminotransferase and aspartate aminotransferase in mice with diabetes is much higher than that of normal mice (see comparison between model group and blank group), but after C ₆₀ - olive oil treatment The values of alanine aminotransferase and aspartate aminotransferase in mice were normal, and there was no significant difference between the olive oil group and the model group, indicating that oil-soluble fullerenes can improve liver function damage caused by diabetes. Figure 5 shows that both urea nitrogen and serum creatinine in diabetic mice are significantly higher than those in normal mice (see comparison between model group and blank group), but urea nitrogen in mice after C ₆₀ - olive oil treatment The serum and creatinine values were normal, and there was no significant difference between the olive oil group and the model group, indicating that oil-soluble fullerenes can improve renal function damage caused by diabetes.

3)尿蛋白检测3) urine protein detection

实验方法：A-D组小鼠给药两周后，将小鼠放于代谢笼中饲养24h，期间收集小鼠的尿液并测量尿液体积，用Elisa试剂盒测试其中尿蛋白的浓度，将所测尿蛋白浓度乘以尿液体积得到24h尿蛋白的排出量。Experimental method: Two weeks after administration of mice in the AD group, the mice were placed in a metabolic cage for 24 hours. The urine of the mice was collected and the volume of urine was measured. The concentration of urinary protein was measured by Elisa kit. The urine protein concentration was multiplied by the urine volume to obtain the 24 hour urine protein discharge.

实验结果：尿蛋白是糖尿病肾病的一个重要指标，图6表明患有糖尿病的小鼠的24h内尿蛋白排出量远远高于正常小鼠的值(参见模型组和空白组的对比)，但是经过C₆₀-橄榄油治疗后的小鼠的尿蛋白排出量显著降低，而橄榄油组与模型组没有明显差异，这说明油溶性富勒烯显著缓解了糖尿病肾病的发展。Experimental results: Urine protein is an important indicator of diabetic nephropathy. Figure 6 shows that the urine output of mice with diabetes is much higher than that of normal mice within 24 hours (see comparison between model group and blank group), but The urine protein excretion of mice treated with C ₆₀ - olive oil was significantly reduced, while the olive oil group was not significantly different from the model group, indicating that oil-soluble fullerenes significantly ameliorated the development of diabetic nephropathy.

4)血管方面表征4) vascular characterization

检测指标：血浆粘度是影响全血粘度的重要因素之一，血浆粘度升高会引起全血粘度增高。Detection index: Plasma viscosity is one of the important factors affecting the viscosity of whole blood. Increased plasma viscosity will cause the whole blood viscosity to increase.

实验方法：给药两周后的A-D组小鼠通过眼球取全血，通过血液流变检测仪在常规条件下检测血液流变性质，Experimental method: A-D mice after two weeks of administration were subjected to whole blood through the eyeball, and the rheological properties of the blood were examined under a normal condition by a blood rheometer.

实验结果：空白组血浆粘度大约为1.5mPa/s，模型组粘度增加为2.0mPa/s，而实验组的粘度明显降低为1.7mPa/s。说明油溶性富勒烯组合物能够减轻糖尿病引起的血液粘度增加等血管疾病。Experimental results: the plasma viscosity of the blank group was about 1.5 mPa/s, the viscosity of the model group was increased to 2.0 mPa/s, and the viscosity of the experimental group was significantly reduced to 1.7 mPa/s. The oil-soluble fullerene composition can alleviate vascular diseases such as an increase in blood viscosity caused by diabetes.

另外通过剪尾巴测量，观察小鼠的状态，可以发现，经过油溶性富勒烯组合物治疗后的小鼠明显伤口愈合加快，说明油溶性富勒烯组合物能够改善糖尿病引起的伤口难以愈合易溃疡等并发症。In addition, by measuring the state of the mouse by shearing the tail, it can be found that the wound healing of the mice after the oil-soluble fullerene composition is accelerated, indicating that the oil-soluble fullerene composition can improve the wounds caused by diabetes and is difficult to heal. Complications such as ulcers.

本发明以C₆₀-橄榄油为实施例对高脂喂养STZ诱导的糖尿病小鼠进行治疗，发现油溶性富勒烯结构对糖尿病并发症有好的缓解作用。The invention uses C ₆₀ - olive oil as an example to treat STZ-induced diabetic mice with high fat feeding, and finds that the oil-soluble fullerene structure has a good alleviating effect on diabetic complications.

实施例4、水溶性内嵌金属富勒烯Gd@C₈₂(OH)_n的体内代谢Example 4: In vivo metabolism of water-soluble inlaid metal fullerene Gd@C ₈₂ (OH) _n

取200μL浓度为1mM的按照实施例2中的方法制备的Gd@C₈₂(OH)_n，将其通过腹腔注射进入C57小鼠体内，24h正常饲养后解剖，取小鼠器官心，肝，脾，肺，肾，胰腺称重，并用65％的硝酸消解在120℃的条件下过夜，稀释50倍后通过ICP测定钆离子浓度。如图7中，可以看出水溶性内嵌金属富勒烯Gd@C₈₂(OH)_n在胰腺富集远多于心、肾、肺和脾。200 μL of Gd@C ₈₂ (OH) _n prepared according to the method of Example 2 was taken at a concentration of 1 mM, and it was intraperitoneally injected into C57 mice. After 24 hours of normal feeding, the mice were dissected and the organs, liver and spleen of the mice were taken. The lungs, kidneys, and pancreas were weighed and digested with 65% nitric acid overnight at 120 ° C. After 50-fold dilution, the cesium ion concentration was measured by ICP. As shown in Figure 7, it can be seen that the water-soluble inlaid metal fullerene Gd@C ₈₂ (OH) _n is enriched in the pancreas much more than the heart, kidney, lung and spleen.

实施例5、水溶性内嵌金属富勒烯Gd@C₈₂(OH)_n清除自由基能力检测Example 5: Water-soluble inlaid metal fullerene Gd@C ₈₂ (OH) _n scavenging free radical ability detection

本发明通过电子自旋共振波谱(ESR)来检测水溶性内嵌金属富勒烯Gd@C₈₂(OH)_n清除自由基的能力。The present invention detects the ability of water-soluble inlaid metal fullerene Gd@C ₈₂ (OH) _{n to} scavenge free radicals by electron spin resonance spectroscopy (ESR).

检测方法：采用紫外诱导产生羟基自由基的方法，将50μL质量浓度为39％的双氧水、50μLPBS缓冲溶液(pH＝7.4)和微量(0.133mM)的二甲基吡啶N-氧化物(DMPO，自由基捕获剂)溶液混合，对照组直接照射280nm的紫外4min，而实验组立即加入200μM的按照实施例2中方法制备的Gd@C₈₂(OH)_n的水溶液10μL后用280nm的紫外光照射4min，检测自由基信号。如图8所示，平滑线为没有加入按照实施例2方法所得的Gd@C₈₂(OH)_n的空白对照，非平滑线为加入Gd@C₈₂(OH)_n的实验组样品，与空白对照相比，加入Gd@C₈₂(OH)_n样品的信号明显降低，说明加入Gd@C₈₂(OH)_n样品的实验组存在的自由基较少，Gd@C₈₂(OH)n有强的清除自由基能力。Detection method: using UV-induced hydroxyl radical generation method, 50 μL of 39% hydrogen peroxide solution, 50 μL PBS buffer solution (pH=7.4) and trace amount (0.133 mM) of lutidine N-oxide (DMPO, free The solution was mixed with the solution, and the control group was directly irradiated with ultraviolet light at 280 nm for 4 min, and the experimental group was immediately added with 10 μL of an aqueous solution of Gd@C ₈₂ (OH) _n prepared according to the method of Example 2, and then irradiated with ultraviolet light of 280 nm for 4 minutes. , detecting free radical signals. As shown, a smooth line is not added according to the method of Example 2. The resulting Gd @ C ₈₂ blank, non-smooth line (OH) _n samples of experimental groups (OH) _n is Gd @ C ₈₂ was added, and the blank 8 Compared with the control, the signal of Gd@C ₈₂ (OH) _n sample was significantly lower, indicating that the experimental group added with Gd@C ₈₂ (OH) _n sample had less free radicals and Gd@C ₈₂ (OH)n was strong. The ability to scavenge free radicals.

实施例6、水溶性内嵌金属富勒烯Gd@C₈₂(OH)_n对糖尿病的治疗Example 6. Treatment of diabetes by water-soluble inlaid metal fullerene Gd@C ₈₂ (OH) _n

(1)实验方法(1) Experimental method

实验动物为db/db糖尿病小鼠，购自南京动物模式中心，引自美国杰克逊实验室。此小鼠是广泛应用的2型糖尿病动物模型，其为瘦素受体基因缺陷导致肥胖后发展为糖尿病的小鼠模型，瘦素受体(leptinreceptor，Lepr)的自发性突变会引起多食、消渴、多尿等症状。db/db小鼠在10～14天出现高胰岛素血症，3～4周明显肥胖，4～8周出现高糖血症。The experimental animals were db/db diabetic mice purchased from the Nanjing Animal Model Center and cited from the Jackson Laboratory in the United States. This mouse is a widely used animal model of type 2 diabetes, which is a mouse model in which the leptin receptor gene defect leads to the development of diabetes after obesity. The spontaneous mutation of the leptin receptor (Lepr) causes polyphagia, Diabetes, polyuria and other symptoms. Db/db mice developed hyperinsulinemia on 10 to 14 days, obesity in 3 to 4 weeks, and hyperglycemia in 4 to 8 weeks.

实验动物分为3组，每组6只。取6只db/m无糖尿病的小鼠作为空白组(简称为A组)，其给药治疗为施用与C组所用药物等体积的生理盐水；随机取6只db/db小鼠作为模型组(简称为B组)，其给药治疗为施用与C组所用药物等体积的生理盐水；随机取6只db/db小鼠作为Gd@C₈₂(OH)_n实验组 (简称为C组)，其给药治疗为施用按照实施例2的方法制备的Gd@C₈₂(OH)_n。A-C组均采用腹腔给药的方式，各组小鼠的周龄进入第10周开始给药，每天给药1次，连续给药两周，C组中Gd@C₈₂(OH)_n的给药剂量为10mg/kg/d。The experimental animals were divided into 3 groups of 6 each. Six db/m non-diabetic mice were used as a blank group (abbreviated as group A), and the administration was performed by administering an equal volume of physiological saline with the drug used in group C; 6 db/db mice were randomly selected as the model group. (referred to as group B), the administration of the drug was the same volume of physiological saline as that used in group C; 6 db/db mice were randomly selected as Gd@C ₈₂ (OH) _n experimental group (referred to as group C) The administration treatment was Gd@C ₈₂ (OH) _n prepared by the method of Example 2. The AC group was administered by intraperitoneal administration. The mice of each group entered the 10th week and started to administer once a day for two weeks. In group C, Gd@C ₈₂ (OH) _n was given. The dose is 10 mg/kg/d.

(2)实验结果(2) Experimental results

本发明通过db/db糖尿病小鼠血糖的变化和糖耐受实验来证明水溶性的内嵌金属富勒烯Gd@C₈₂(OH)_n对糖尿病的治疗作用，通过血生化证明其改善体内氧化还原水平。The present invention demonstrates the therapeutic effect of water-soluble inlaid metal fullerene Gd@C ₈₂ (OH) _n on diabetes by db/db diabetes mellitus changes in blood glucose and glucose tolerance test, and improves blood oxidation in vivo by blood biochemistry. Restore level.

1)空腹血糖1) Fasting blood sugar

第1次给药前测空腹血糖，计为0d，之后在第5天、第9天和第14天给药前测空腹血糖，每次测空腹血糖前需禁食3h。结果如图9所示，图9为空腹血糖变化图，从图9中可以看出模型组中只注射生理盐水进行治疗的糖尿病小鼠在第14天时的空腹血糖要远高于实验组注射Gd@C₈₂(OH)_n进行治疗的糖尿病小鼠，也远高于空白组中无糖尿病的小鼠的空腹血糖，而实验组注射Gd@C₈₂(OH)_n进行治疗的糖尿病小鼠的空腹血糖趋于正常。Fasting blood glucose was measured before the first administration, which was counted as 0d, and then fasting blood glucose was measured before administration on the 5th, 9th, and 14th days, and fasting was required for 3 hours before the fasting blood glucose was measured. The results are shown in Fig. 9. Fig. 9 is a graph showing changes in fasting blood glucose. It can be seen from Fig. 9 that the fasting blood glucose of the diabetic mice treated with saline only in the model group is much higher than that of the experimental group. The diabetic mice treated with @C ₈₂ (OH) _n were also much higher than the fasting blood glucose of the non-diabetic mice in the blank group, while the experimental group was fasted to the diabetic mice treated with Gd@C ₈₂ (OH) _n . Blood sugar tends to be normal.

2)糖耐受实验2) Sugar tolerance test

给药2周结束后进行糖耐受实验。禁食12h后测血糖值计为0min的血糖值，然后灌胃葡萄糖溶液，剂量为2g/kg小鼠体重，然后分别在15，30，60，120min时测血糖，作图得到糖耐受曲线，并且计算了曲线下面积(AUC)来表征小鼠的糖耐受能力。结果如图10和图11所示，实验组注射Gd@C₈₂(OH)_n进行治疗的糖尿病小鼠对血糖的控制能力强于模型组只注射生理盐水进行治疗的糖尿病小鼠，且实验组的AUC明显小于模型组，说明Gd@C₈₂(OH)_n能够较好地治疗糖尿病，增加糖耐量，增强小鼠对血糖的调节控制能力。Glucose tolerance test was performed after 2 weeks of administration. After fasting for 12 hours, the blood glucose level was measured as 0 min blood glucose, then the glucose solution was administered at a dose of 2 g/kg of mouse body weight, and then blood glucose was measured at 15, 30, 60, and 120 min, respectively, and the glucose tolerance curve was obtained. And the area under the curve (AUC) was calculated to characterize the glucose tolerance of the mice. Results As shown in Fig. 10 and Fig. 11, the diabetic mice treated with Gd@C ₈₂ (OH) _n in the experimental group had better control of blood glucose than the diabetic mice treated with normal saline in the model group, and the experimental group The AUC was significantly smaller than that of the model group, indicating that Gd@C ₈₂ (OH) _n can better treat diabetes, increase glucose tolerance, and enhance the ability of mice to regulate and control blood sugar.

3)体内氧化还原水平测试3) In vivo redox level test

CAT是一种酶类清除剂，它可促使H₂O₂分解为分子氧和水，清除体内的过氧化氢，从而使细胞免于遭受H₂O₂的毒害。MDA为自由基代谢产物，产生更多的自由基时，其含量会增加。CAT is an enzyme scavenger that decomposes H ₂ O ₂ into molecular oxygen and water, removing hydrogen peroxide from the body, thereby protecting cells from H ₂ O ₂ poisoning. MDA is a free radical metabolite, and its content increases when more free radicals are produced.

给药两周后的各组小鼠通过眼球取血，将血液样品在室温下静置1h后，3500rpm离心15min，吸取上层血清，利用分光光度法，测定405nm处的吸光度计算得到过氧化氢酶(CAT)的含量，测定532nm处的吸光度计算得到丙二醛(MDA)的含量，以上2种物质的检测均可通过市面可获得的相应试剂盒进行检测。如图12所示Gd@C₈₂(OH)n治疗后的小鼠体内过氧化氢酶增加，如图13所示Gd@C₈₂(OH)n治疗后的小鼠体内丙二醛减少，说明自由基减少，这两个指标都说明水溶性内嵌金属富勒烯治疗糖尿病不仅降低血糖，更重要的是会改善小鼠体内的氧化还原水平，减少并发症的发生。Two weeks after the administration, the mice in each group were bled by the eyeball, and the blood sample was allowed to stand at room temperature for 1 hour, centrifuged at 3500 rpm for 15 min, and the upper serum was aspirated, and the absorbance at 405 nm was measured by spectrophotometry to obtain catalase. The content of (CAT) is determined by measuring the absorbance at 532 nm to obtain the content of malondialdehyde (MDA), and the detection of the above two substances can be detected by a commercially available corresponding kit. As shown in FIG _{Gd @ C 82 (OH) 12} n mice after treatment catalase increases as Gd @ C ₈₂ (OH) n to reduce the in vivo treatment of mice MDA shown in FIG. 13, described Free radical reduction, both indicators indicate that water-soluble inlaid metal fullerene treatment of diabetes not only reduces blood sugar, but more importantly, it improves the redox level in mice and reduces the occurrence of complications.

上述实施例对基因缺陷db/db糖尿病小鼠进行了治疗，结果表明水溶性的内嵌金属富勒烯Gd@C₈₂(OH)_n具有降血糖、增加糖耐量的功能，并且还能够改善小鼠体内的氧化还原水平，从而能从根本上治疗糖尿病引起的肝肾损伤、心血管病等并发症。The above examples were used to treat genetically deficient db/db diabetic mice. The results showed that the water-soluble inlaid metal fullerene Gd@C ₈₂ (OH) _n has the functions of lowering blood sugar, increasing glucose tolerance, and also improving small The level of redox in the mouse can fundamentally treat complications such as liver and kidney damage and cardiovascular disease caused by diabetes.

实施例7、水溶性富勒烯或油溶性富勒烯对胰岛素抵抗的治疗Example 7. Treatment of insulin resistance by water-soluble fullerene or oil-soluble fullerene

(1)实验方法(1) Experimental method

实验动物为db/db糖尿病小鼠，购自南京动物模式中心，引自美国杰克逊实验室。此小鼠为瘦素受体基因缺陷导致肥胖后发展成2型糖尿病的小鼠模型，db/db小鼠在10～14天出现高胰岛素血症，3～4周明显肥胖，4～8周出现高糖血症。其具有2型糖尿病的特点，会产生典型的胰岛素抵抗。The experimental animals were db/db diabetic mice purchased from the Nanjing Animal Model Center and cited from the Jackson Laboratory in the United States. This mouse is a mouse model of leptin receptor gene deficiency leading to development of type 2 diabetes after obesity. db/db mice develop hyperinsulinemia on 10 to 14 days, and obesity in 3 to 4 weeks, 4 to 8 weeks. Hyperglycemia occurs. It has the characteristic of type 2 diabetes and produces typical insulin resistance.

实验动物分为4组，每组6只。取6只db/m无糖尿病的小鼠作为空白组(简称为A组)，其给药治疗为施用与C组药物同体积的生理盐水；随机取6只db/db小鼠作为模型组(简称为B组)，其给药治疗为施用与C组药物同体积的生理盐水；随机取6只db/db小鼠作为C₇₀(OH)_n实验组(简称为C组)，其给药治疗为施用按照实施例2的方法制备的C₇₀(OH)_n；随机取6只db/db小鼠作为C₆₀-橄榄油实验组(简称为D组)，其给药治疗为施用按照实施例1的方法制备的C₆₀-橄榄油；ABC组均采用腹腔给药的方式，D组采用灌胃给药的方式，各组小鼠的周龄进入第10周开始给药，每天给药1次，连续给药两周，C组和D组的给药剂量为10mg/kg/d。The experimental animals were divided into 4 groups of 6 each. Six db/m non-diabetic mice were used as the blank group (abbreviated as group A), and the administration was performed by administering the same volume of physiological saline as the group C drug; 6 db/db mice were randomly taken as the model group ( Referred to as group B), the administration of the drug is the same volume of physiological saline as that of group C; 6 db/db mice are randomly taken as the C ₇₀ (OH) _n experimental group (referred to as group C), and the drug is administered. The treatment was to apply C ₇₀ (OH) _n prepared according to the method of Example 2; 6 db/db mice were randomly taken as the C ₆₀ - olive oil experimental group (abbreviated as group D), and the administration treatment was performed according to the administration. The C ₆₀ - olive oil prepared by the method of Example 1; the ABC group was administered by intraperitoneal administration, and the D group was administered by intragastric administration. The mice of each group entered the 10th week and started to be administered daily. One time, continuous administration for two weeks, and the doses of groups C and D were 10 mg/kg/d.

(2)实验结果(2) Experimental results

1)胰岛素耐受实验1) Insulin tolerance test

在给药2周后进行胰岛素耐受实验。禁食4h后测量血糖，计为0分钟的血糖值，然后灌胃给胰岛素，剂量为1U/kg小鼠体重，然后灌胃后的15，30，60，120min时分别测量血糖，作图得到胰岛素耐受曲线。结果如图14所示，注射入胰岛素后，空白组中无糖尿病的小鼠的血糖在前15min内下降最快；而模型组中具有糖尿病但只接受生理盐水治疗的db/db糖尿病小鼠在前15min 内血糖几乎没有下降，整个120min的过程中下降的平均速度也很缓慢；与模型组相比，C₇₀(OH)_n实验组和C₆₀-橄榄油实验组的db/db糖尿病小鼠血糖都有所降低，且血糖降低的平均速度也明显快于模型组，尤其是前15min，因此证明施用C₇₀(OH)_n和C₆₀-橄榄油后，使患有糖尿病的小鼠打入胰岛素后可以降血糖，并且对胰岛素的敏感性增加，对胰岛素的抵抗减弱。The insulin resistance test was performed 2 weeks after the administration. After fasting for 4 hours, the blood glucose was measured, and the blood glucose level was counted as 0 minutes. Then, the insulin was administered to the patient at a dose of 1 U/kg of the mouse body weight, and then the blood glucose was measured at 15, 30, 60, and 120 minutes after the gavage. Insulin tolerance curve. The results are shown in Figure 14. After injection of insulin, the blood glucose of the non-diabetic mice in the blank group decreased the fastest in the first 15 min; while the db/db diabetic mice in the model group who had diabetes but received only saline treatment were There was almost no drop in blood glucose during the first 15 minutes, and the average rate of decline during the entire 120-min period was also slow; compared with the model group, db/db diabetic mice in the C ₇₀ (OH) _n experimental group and the C ₆₀ -olive oil experimental group Blood sugar was reduced, and the average rate of blood glucose reduction was also significantly faster than the model group, especially the first 15 min, thus demonstrating that mice with diabetes were injected after administration of C ₇₀ (OH) _n and C ₆₀ - olive oil. After insulin can lower blood sugar, and the sensitivity to insulin increases, and the resistance to insulin is weakened.

2)甘油三酯和胆固醇的含量检测2) Detection of triglyceride and cholesterol

有研究报道，高甘油三酯和高胆固醇可作为胰岛素抵抗的表现。Studies have reported that high triglycerides and high cholesterol can be used as a manifestation of insulin resistance.

给药2周后，小鼠通过眼球取血，室温静置1h后，3500rpm离心15min，吸取上层血清，通过自动血生化仪测定甘油三酯TG、总胆固醇TC。结果如图15所述，模型组的甘油三酯和总胆固醇相对于空白组显著上升，而施用C₇₀(OH)_n和C₆₀-橄榄油后的小鼠的甘油三酯和胆固醇的含量相对于模型组则明显的降低且趋于正常。表明C₇₀(OH)_n和C₆₀-橄榄油可使小鼠对胰岛素的敏感性增加，对胰岛素的抵抗减弱。Two weeks after the administration, the mice were bled by eyeballs, allowed to stand at room temperature for 1 hour, centrifuged at 3500 rpm for 15 min, and the supernatant serum was aspirated, and triglyceride TG and total cholesterol TC were measured by an automatic blood biochemistry analyzer. As a result, as shown in Fig. 15, the triglyceride and total cholesterol of the model group were significantly increased relative to the blank group, whereas the contents of the triglyceride and cholesterol of the mice after administration of _C70 (OH) _n and _C60 -olive oil were relatively In the model group, it is obviously reduced and tends to be normal. It is shown that C ₇₀ (OH) _n and C ₆₀ - olive oil can increase the sensitivity of mice to insulin and the resistance to insulin is weakened.

3)葡萄糖转运蛋白-4的相对含量检测3) Detection of relative content of glucose transporter-4

大量研究已证明2型糖尿病的主要病理生理特征是胰岛素抵抗，其主要病理改变之一是指脂肪细胞和骨骼肌细胞内胰岛素信号通路受损致葡萄糖转运蛋白GLUT4转位障碍。即胰岛素抵抗会引起葡萄糖转运蛋白-4含量降低。A large number of studies have proved that the main pathophysiological feature of type 2 diabetes is insulin resistance. One of the main pathological changes is that the insulin signaling pathway in adipocytes and skeletal muscle cells is impaired and the glucose transporter GLUT4 translocation disorder. That is, insulin resistance causes a decrease in the content of glucose transporter-4.

给药2周后，取小鼠肩胛处棕色脂肪冻存，研成组织匀浆，通过Weratern Blot的方法测试脂肪中葡萄糖转运蛋白-4的含量。如图17所示，模型组的葡萄糖转运蛋白-4相对于空白组显著下降，而施用C₇₀(OH)_n和C₆₀-橄榄油后的小鼠的脂肪细胞中的葡萄糖转运蛋白-4的含量明显增加，表明C₇₀(OH)_n和C₆₀-橄榄油可使小鼠对胰岛素的敏感性增加，对胰岛素的抵抗减弱。Two weeks after the administration, the brown fat in the shoulder of the mouse was frozen, and the tissue homogenate was ground, and the content of glucose transporter-4 in the fat was measured by the method of Weratern Blot. As shown in Figure 17, the glucose transporter-4 of the model group was significantly decreased relative to the blank group, whereas the glucose transporter-4 in the adipocytes of mice after administration of _C70 (OH) _n and _C60 -olive oil The content increased significantly, indicating that C ₇₀ (OH) _n and C ₆₀ - olive oil increased the sensitivity of mice to insulin and decreased resistance to insulin.

4)胰岛素敏感指数4) Insulin sensitivity index

给药2周后，禁食12h测定小鼠的空腹血糖。并取血2mL低温离心，分离血清，以放射免疫竞争法测定血清胰岛素水平(单纯性肥胖：基础与临床【M】.北京：北京科学技术出版社，1998:2)，计算胰岛素敏感指数ISI，公式为ISI＝ln(1/(空腹血糖*空腹胰岛素))。结果如下表，施用C₇₀(OH)_n和C₆₀-橄榄油后，胰岛素敏感指数增加，也说明C₇₀(OH)_n和C₆₀-橄榄油后能够使小鼠对胰岛素的敏感性增加，对胰岛素的抵抗减弱。Two weeks after the administration, the fasting blood glucose of the mice was measured by fasting for 12 hours. Blood samples were taken from 2 mL of low temperature, serum was separated, and serum insulin levels were determined by radioimmunoassay (simple obesity: basic and clinical [M]. Beijing: Beijing Science and Technology Press, 1998: 2), and the insulin sensitivity index ISI was calculated. The formula is ISI=ln (1/(fasting blood glucose* fasting insulin)). The results are shown in the table below. After administration of C ₇₀ (OH) _n and C ₆₀ - olive oil, the insulin sensitivity index increased, indicating that C ₇₀ (OH) _n and C ₆₀ - olive oil can increase the sensitivity of mice to insulin. The resistance to insulin is weakened.

本发明以C₆₀-橄榄油组合物和C₇₀(OH)_n为例对db/db小鼠进行了治疗，发现水溶性富勒烯和油溶性富勒烯能使小鼠对胰岛素的敏感性增加，对胰岛素的抵抗减弱。The present invention treats db/db mice with C ₆₀ - olive oil composition and C ₇₀ (OH) _n as an example, and found that water-soluble fullerenes and oil-soluble fullerenes can make mice sensitive to insulin. Increased, the resistance to insulin is weakened.

前述对本发明的具体示例性实施方案的描述是为了说明和例证的目的。这些描述并非想将本发明限定为所公开的精确形式，并且很显然，根据上述教导，可以进行很多改变和变化。对示例性实施例进行选择和描述的目的在于解释本发明的特定原理及其实际应用，从而使得本领域的技术人员能够实现并利用本发明的各种不同的示例性实施方案以及各种不同的选择和改变。本发明的范围意在由权利要求书及其等同形式所限定。 The foregoing description of the specific exemplary embodiments of the present invention has The description is not intended to limit the invention to the precise forms disclosed. The embodiments were chosen and described in order to explain the particular embodiments of the invention Choose and change. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

一种富勒烯结构在制备治疗糖尿病及其并发症的药物中的应用，其特征在于：所述富勒烯结构包括至少一种选自下组的有效成分：油溶性的富勒烯、油溶性的内嵌金属富勒烯、所述油溶性的富勒烯和所述油溶性的内嵌金属富勒烯的组合物、水溶性的富勒烯、水溶性的内嵌金属富勒烯、所述水溶性的富勒烯和所述水溶性的内嵌金属富勒烯的组合物、以上六者的可药用的酯或以上六者的可药用的盐。Use of a fullerene structure for the preparation of a medicament for treating diabetes and its complications, characterized in that the fullerene structure comprises at least one active ingredient selected from the group consisting of oil-soluble fullerenes and oils a soluble inlaid metal fullerene, a composition of the oil-soluble fullerene and the oil-soluble inlaid metal fullerene, a water-soluble fullerene, a water-soluble inlaid metal fullerene, A composition of the water-soluble fullerene and the water-soluble inlaid metal fullerene, a pharmaceutically acceptable ester of the above six or a pharmaceutically acceptable salt of the above six.
一种治疗糖尿病及其并发症的药物组合物，其特征在于：包括至少一种选自下组的富勒烯结构：油溶性的富勒烯、油溶性的内嵌金属富勒烯、所述油溶性的富勒烯和所述油溶性的内嵌金属富勒烯的组合物、水溶性的富勒烯、水溶性的内嵌金属富勒烯、所述水溶性的富勒烯和所述水溶性的内嵌金属富勒烯的组合物、以上六者的可药用的酯、以上六者的可药用的盐；所述药物组合物还包括可药用的载体、可药用的稀释剂和可药用的赋形剂中的至少一种。A pharmaceutical composition for treating diabetes and its complications, comprising: at least one fullerene structure selected from the group consisting of oil-soluble fullerenes, oil-soluble inlaid metal fullerenes, said a composition of an oil-soluble fullerene and the oil-soluble inlaid metal fullerene, a water-soluble fullerene, a water-soluble inlaid metal fullerene, the water-soluble fullerene, and the a water-soluble composition of a metal fullerene, a pharmaceutically acceptable ester of the above six, a pharmaceutically acceptable salt of the above six; the pharmaceutical composition further comprising a pharmaceutically acceptable carrier, pharmaceutically acceptable At least one of a diluent and a pharmaceutically acceptable excipient.
一种治疗糖尿病及其并发症的方法，包括向需要治疗糖尿病及其并发症的受试者施用有效量的富勒烯结构，所述富勒烯结构包括至少一种选自下组的有效成分：油溶性的富勒烯、油溶性的内嵌金属富勒烯、所述油溶性的富勒烯和所述油溶性的内嵌金属富勒烯的组合物、水溶性的富勒烯、水溶性的内嵌金属富勒烯、所述水溶性的富勒烯和所述水溶性的内嵌金属富勒烯的组合物、以上六者的可药用的酯或以上六者的可药用的盐。A method of treating diabetes and its complications, comprising administering to a subject in need of treatment for diabetes and a complication thereof an effective amount of a fullerene structure comprising at least one active ingredient selected from the group consisting of : oil-soluble fullerene, oil-soluble inlaid metal fullerene, oil-soluble fullerene and oil-soluble inlaid metal fullerene composition, water-soluble fullerene, water-soluble a medicinal embedded fullerene, a water-soluble fullerene and a water-soluble inlaid metal fullerene composition, a pharmaceutically acceptable ester of the above six or a medicinal of the above six Salt.
根据权利要求1所述的应用或权利要求2所述的药物组合物或权利要求3所述的方法，其特征在于：所述油溶性的富勒烯包括碳笼外表面包覆有油溶液的富勒烯，所述油溶性的内嵌金属富勒烯包括碳笼外表面包覆有油溶液的内嵌金属富勒烯。The pharmaceutical composition according to claim 1 or the method according to claim 2 or the method according to claim 3, wherein the oil-soluble fullerene comprises an outer surface of the carbon cage coated with an oil solution Fullerene, the oil-soluble inlaid metal fullerene comprises an inlaid metal fullerene having an outer surface of a carbon cage coated with an oil solution.
根据权利要求4所述的应用或权利要求4所述的药物组合物或权利要求4所述的方法，其特征在于：所述油溶液包括橄榄油、亚麻籽油、葵花籽油、玉米胚油、角鲨烷中的至少一种。The pharmaceutical composition according to claim 4 or the method according to claim 4 or the method according to claim 4, wherein the oil solution comprises olive oil, linseed oil, sunflower oil, corn germ oil At least one of squalane.
根据权利要求1所述的应用或权利要求2所述的药物组合物或权利要求3所述的方法，其特征在于：所述水溶性的富勒烯包括选自下组的至少一种富勒烯：1)碳笼外表面修饰有亲水基团的富勒烯；(2)碳笼外表面被亲水性生物小分子包裹的富勒烯；(3)被具有生物相容性的载体材料负载的富勒烯；(4)自组装形成的水溶性超分子体系富勒烯。The pharmaceutical composition according to claim 1 or the pharmaceutical composition according to claim 2 or the method according to claim 3, wherein the water-soluble fullerene comprises at least one fuller selected from the group consisting of Aene: 1) fullerene modified with a hydrophilic group on the outer surface of the carbon cage; (2) the outer surface of the carbon cage is hydrophilic a fullerene coated with a small molecule of a biological organism; (3) fullerenes supported by a biocompatible carrier material; (4) a water-soluble supramolecular system fullerene formed by self-assembly.
根据权利要求1所述的应用或权利要求2所述的药物组合物或权利要求3所述的方法，其特征在于：所述水溶性的内嵌金属富勒烯包括选自下组的至少一种内嵌金属富勒烯：(1)碳笼外表面修饰有亲水基团的内嵌金属富勒烯；(2)碳笼外表面被亲水性生物小分子包裹的内嵌金属富勒烯；(3)被具有生物相容性的载体材料负载的内嵌金属富勒烯；(4)自组装形成的水溶性超分子体系内嵌金属富勒烯。The pharmaceutical composition according to claim 1 or the pharmaceutical composition according to claim 2 or the method according to claim 3, wherein the water-soluble inlaid metal fullerene comprises at least one selected from the group consisting of Inlaid metal fullerenes: (1) inlaid metal fullerenes modified with a hydrophilic group on the outer surface of the carbon cage; (2) inlaid metal fullerene surrounded by hydrophilic biological small molecules on the outer surface of the carbon cage (3) embedded metal fullerenes supported by a biocompatible carrier material; (4) a metal-filled fullerene embedded in a water-soluble supramolecular system formed by self-assembly.
根据权利要求6-7所述的应用或权利要求6-7所述的药物组合物或权利要求6-7所述的方法，其特征在于：所述亲水基团包括羟基、羧基、巯基和氨基中的一种或多种。The use according to claims 6-7 or the pharmaceutical composition according to claims 6-7 or the method according to claims 6-7, wherein the hydrophilic group comprises a hydroxyl group, a carboxyl group, a thiol group and One or more of the amino groups.
根据权利要求1所述的应用或权利要求2所述的药物组合物或权利要求3所述的方法，其特征在于：所述油溶性的富勒烯是通过将原料富勒烯进行油溶性改性获得的；所述油溶性的内嵌金属富勒烯是通过将原料内嵌金属富勒烯进行油溶性改性获得的；所述水溶性的富勒烯是通过将原料富勒烯进行水溶性改性获得的；所述水溶性的内嵌金属富勒烯是通过将原料内嵌金属富勒烯进行水溶性改性获得的。The pharmaceutical composition according to claim 1 or the pharmaceutical composition according to claim 2 or the method according to claim 3, wherein the oil-soluble fullerene is modified by oil-soluble fullerene Obtained; the oil-soluble inlaid metal fullerene is obtained by oil-soluble modification of the raw material inlaid with metal fullerenes; the water-soluble fullerene is water-soluble by the fullerene Obtained by the modification; the water-soluble inlaid metal fullerene is obtained by water-soluble modification by embedding metal fullerenes in the raw material.
根据权利要求1所述的应用或权利要求2所述的药物组合物或权利要求3所述的方法，其特征在于：所述原料富勒烯包括一种或多种通式为C_2m的由碳原子组成的笼状结构，30≤m≤60；所述内嵌金属富勒烯原料包括M@C_2n、M₂@C_2n、MA@C_2n、M₃N@C_2n、M₂C₂@C_2n、M₂S@C_2n、M₂O@C_2n和M_xA_3-xN@C2n中的一种或多种，其中：M、A均代表金属元素且M、A均选自Sc、Y和镧系金属元素中的任意一种，30≤n≤60；0≤x≤3；N代表氮元素，C代表碳元素，S代表硫元素，镧系金属元素包括La、Ce、Pr、Nd、Pm、Sm、Eu、Gd、Tb、Dy、Ho、Er、Tm、Yb和Lu。The pharmaceutical composition according to claim 1 or the pharmaceutical composition according to claim 2 or the method according to claim 3, wherein the raw fullerene comprises one or more of the formula C _2m a cage structure composed of carbon atoms, 30 ≤ m ≤ 60; the embedded metal fullerene raw material includes M@C _2n , M ₂ @C _2n , MA@C _2n , M ₃ N@C _2n , M ₂ C _{2 one or more of} @C _2n , M ₂ S@C _2n , M ₂ O@C _2n and M _x A _3-x N@C2n, wherein: M and A both represent metal elements and M and A are both Any one selected from the group consisting of Sc, Y and lanthanide metal elements, 30 ≤ n ≤ 60; 0 ≤ x ≤ 3; N represents a nitrogen element, C represents a carbon element, S represents a sulfur element, and a lanthanide metal element includes La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu.
根据权利要求1所述的应用或权利要求2所述的药物组合物或权利要求3所述的方法，其特征在于：所述糖尿病为一型糖尿病或二型糖尿病。The pharmaceutical composition according to claim 1 or the pharmaceutical composition according to claim 2 or the method according to claim 3, wherein the diabetes is type 1 diabetes or type 2 diabetes.
根据权利要求1所述的应用或权利要求2所述的药物组合物或权利要求3所述的方法，其特征在于：所述糖尿病并发症包括糖尿病性心脑血管病，可选的为微血管和/或大血管病变引起的心脑血管疾病，如：动脉粥样硬化；糖尿病肾病；糖尿病皮肤病，如：糖尿病溃疡，伤口愈合困难；糖尿病引起的高粘稠血症；糖尿病神经病变，如：中风；糖尿病眼部并发症，如：视网膜病变；糖尿病病足。The pharmaceutical composition according to claim 1 or the method according to claim 2 or the method according to claim 3, wherein the diabetic complications include diabetic cardio-cerebral vascular disease, optionally microvessels and / or cardiovascular disease caused by macrovascular disease, such as: atherosclerosis Diabetic nephropathy; diabetic skin disease, such as: diabetic ulcer, difficulty in wound healing; hyperviscosity caused by diabetes; diabetic neuropathy, such as: stroke; diabetic eye complications, such as: retinopathy; diabetic foot.
根据权利要求1所述的应用或权利要求2所述的药物组合物或权利要求3所述的方法，其特征在于：所述治疗糖尿病并发症包括：1)使糖尿病引起的血糖升高趋于正常；2)使胰岛的大小和胰岛细胞的数量趋于正常；3)调节胰岛素分泌，减轻胰岛素抵抗，增加胰岛素敏感性，使糖耐量和胰岛素耐量趋于正常；4)治疗糖尿病引起的冠心病；5)使糖尿病肾病引起的肝肾指标失常趋于正常(肝肾指标包括：谷丙转氨酶，谷草转氨酶，血肌酐，尿素氮)，使尿蛋白明显降低；6)使伤口愈合加快；7)降低糖尿病引起的血液粘度增加；8)降低血脂(血脂指标包括：总胆固醇，甘油三酯，高低密度脂)；9)治疗糖尿病病足；10)治疗糖尿病性视网膜病变、与糖尿病相关的葡萄膜炎和糖尿病性白内障；11)使糖化血红蛋白显著降低；12)使糖基化产物(AGE)降低；13)调节机体免疫及炎症；14)改善血液流变异常，抑制血小板高聚集性；15)预防及改善神经功能不全。 The pharmaceutical composition according to claim 1 or the method according to claim 2 or the method according to claim 3, wherein the treating diabetic complications comprises: 1) stimulating blood sugar elevation caused by diabetes Normal; 2) make the size of islets and the number of islet cells tend to be normal; 3) regulate insulin secretion, reduce insulin resistance, increase insulin sensitivity, make glucose tolerance and insulin tolerance tend to normal; 4) treat coronary heart disease caused by diabetes 5) The liver and kidney indicators caused by diabetic nephropathy tend to be normal (hepatic and renal indicators include: alanine aminotransferase, aspartate aminotransferase, serum creatinine, urea nitrogen), so that the urine protein is significantly reduced; 6) accelerate wound healing; 7) Reduce blood viscosity increase caused by diabetes; 8) Reduce blood lipids (blood lipid indicators include: total cholesterol, triglycerides, high and low density lipids); 9) treat diabetic foot; 10) treat diabetic retinopathy, diabetes-related grapes Membrane inflammation and diabetic cataract; 11) Significantly reduce glycated hemoglobin; 12) Reduce glycosylation products (AGE); 13) Regulate immune and inflammation; 14) Change Abnormal blood rheology, high inhibition of platelet aggregation; 15) prevent and improve neurological dysfunction.