CN103191417A - Long-effect blood-glucose-control medicine-controlled-release system loading insulin aggregate, and preparation method thereof - Google Patents

Long-effect blood-glucose-control medicine-controlled-release system loading insulin aggregate, and preparation method thereof Download PDF

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CN103191417A
CN103191417A CN 201210002190 CN201210002190A CN103191417A CN 103191417 A CN103191417 A CN 103191417A CN 201210002190 CN201210002190 CN 201210002190 CN 201210002190 A CN201210002190 A CN 201210002190A CN 103191417 A CN103191417 A CN 103191417A
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insulin
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李建树
罗珺
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四川大学
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Abstract

The invention relates to a long-effect blood-glucose-control medicine-controlled-release system loading insulin aggregate. The system is composed of a substrate, and a polymer layer with pH sensitivity and an insulin aggregate layer superposed or coated on the substrate, wherein the substrate, the polymer layer with pH sensitivity, and the insulin aggregate layer are integrated through an electrostatic effect. Or, the system is composed of a substrate, a polymer layer with pH sensitivity, a protein layer, and an insulin aggregate layer superposed or coated on the substrate, wherein the substrate, the polymer layer with pH sensitivity, the protein layer, and the insulin aggregate layer are integrated through an electrostatic effect. A preparation method of the invention comprises the steps that: 1, a raw material solution or suspension liquid is prepared; 2, a substrate surface is treated or a substrate is prepared; and 3, the coating layer or an encapsulation layer is assembled on the substrate.

Description

载有胰岛素聚集体的长效控制血糖浓度的药物控释体系及 Containing long acting insulin aggregates of blood glucose concentration control system and controlled drug release

制备方法 Preparation

技术领域 FIELD

[0001] 本发明属于生物材料和药剂学交叉的技术领域,特别涉及可储存和控制胰岛素聚集体释放的体系及制备方法。 [0001] The present invention belongs to the technical field of biological and pharmaceutical materials crossing, and more particularly to system and method for preparing a control store and release insulin aggregates.

背景技术 Background technique

[0002] 研究显示,对糖尿模型动物单次皮下注射牛胰岛素聚集体(简称B-SIA) 200 μ g,可以控制其体内血糖浓度正常120天,对糖尿病模型动物单次皮下注射重组人胰岛素聚集体(简称rH-SIA)200yg,可以控制体其内血糖浓度正常140天,但对糖尿病模型动物单次皮下注射胰岛素聚集体400 μ g,则会导致过度低血糖(见Gupta, S., Chattopadhyay, T.,Singh,MP,et al.,Supramolecular insulin assembly II for a sustained treatmentof type I diabetes mellitus.Proc.Natl.Acad.Sc1.USA 2010,107:13246-13251.);对糖尿病模型动物单次皮下注射猪胰岛素聚集体(简称P_SIA)200yg,可以控制其体内血糖浓度正常129天。 [0002] Studies show, diabetes model animals of a single subcutaneous injection of bovine insulin aggregates (referred to as B-SIA) 200 μ g, can control their blood sugar levels were normal 120 days, diabetic model animals a single subcutaneous injection of recombinant human insulin aggregates body (referred to as rH-SIA) 200yg, blood glucose concentration within the normal control body may be 140 days, but for a single subcutaneous injection diabetic model animal insulin accumulation body 400 μ g will cause excessive hypoglycemia (see Gupta, S., Chattopadhyay , T., Singh, MP, et al, Supramolecular insulin assembly II for a sustained treatmentof type I diabetes mellitus.Proc.Natl.Acad.Sc1.USA 2010,107:.. 13246-13251); diabetic model animals single subcutaneous porcine insulin aggregates (referred P_SIA) 200yg, can control their blood sugar levels were normal 129 days. 研究还显示,由于各种原因(例如缺乏智能响应性的控制释放)导致的胰岛素剂量控制不当会极大增加各种癌症的发病率和致死率(见Smith,U.,Gale,EAM,Does diabetes therapy influence the risk of cancer ? Diabetologia 2009,52:1699-1708.)。 The study also showed that insulin doses due to various reasons (such as lack of intelligent response of the controlled release) resulting from improper control will greatly increase morbidity and mortality rates of various cancers (see Smith, U., Gale, EAM, Does diabetes therapy influence the risk of cancer Diabetologia 2009,52:? 1699-1708).. 病例报道显示,长期直接使用胰岛素的部位有可能产生胰岛素的沉积而导致淀粉样变形病(见Dische, FE,ffernstedt, C.,ffestermark, GT,et al.,Insulin asan amyloid-fibril protein at sites of repeated insulin injection in a diabeticpatient.Diabetologia 1988,31:158-161.)。 Case reports indicate that long-term use of insulin directly to the site of deposition of the insulin may occur resulting in deformation of amyloid disease (see Dische, FE, ffernstedt, C., ffestermark, GT, et al., Insulin asan amyloid-fibril protein at sites of repeated insulin injection in a diabeticpatient.Diabetologia 1988,31: 158-161).. 因而,非常有必要构建能负载并控制胰岛素聚集体释放的智能响应性系统。 Accordingly, it is necessary to construct a great load and the intelligent control system responsive insulin release aggregate.

发明内容 SUMMARY

[0003] 本发明的目的在于克服现有技术的不足,提供载有胰岛素聚集体的长效控制血糖浓度的药物控释体系及制备方法,所述药物控释体系不仅可延长糖尿病模型动物体内血糖浓度正常的时间,而且可智能响应性控制所载胰岛素的释放。 [0003] The object of the present invention is to overcome the disadvantages of the prior art, and provide a method for preparing a pharmaceutical controlled release system containing the long-acting insulin aggregates of blood glucose concentration control, the system is not only prolong the drug release in vivo glucose diabetic model animals the concentration of the normal time, but also the release of insulin in response to control intelligence contained.

[0004] 本发明所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,有下述两种类型。 [0004] The present invention contains a drug release depot system control of blood glucose concentrations of insulin aggregates, there are the following two types.

[0005] 1、第一种类型:基底为非胰岛素聚集体 [0005] 1, the first type: non-insulin-aggregate substrate

[0006] 此种类型的药物控释体系由基底及重叠或包覆在基底上的具有pH敏感性的聚合物层和胰岛素聚集体层组成,基底、具有pH敏感性的聚合物层、胰岛素聚集体层相互之间通过静电作用结合成一体;或由基底及重叠或包覆在基底上的具有PH敏感性的聚合物层、蛋白质层和胰岛素聚集体层组成,基底、具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层相互之间通过静电作用结合成一体;所述基底为平板、微球或医疗器械,所述平板或微球由有机玻璃、石英、云母中的一种制作。 [0006] This type of controlled drug delivery systems by the substrate and overlapping or covering polymeric layer on the substrate layer and the aggregation of insulin having a pH sensitive composition, the substrate, having a pH-sensitive polymer layer, aggregation of insulin between the layers bonded to each other integrally by electrostatic interaction; or overlapping or by a base and coated with a polymer layer on a substrate having a sensitive PH, the protein layer and insulin aggregate layers, the substrate, having a PH sensitive polymerization layer, a protein layer, between insulin aggregate layer electrostatically bonded to each other integrally; said substrate is a tablet, microspheres or medical device, the microspheres are made of a plate or organic glass, quartz, mica prepared .

[0007] 上述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,具有pH敏感性的聚合物层和胰岛素聚集体层在基底上的包覆顺序可以是具有pH敏感性的聚合物层、胰岛素聚集体层的所有组合方式,或具有PH敏感性的聚合物层、蛋白质层和胰岛素聚集体层在基底上的包覆顺序可以是具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层的所有组合方式。 [0007] The long-term control of blood glucose concentration of the drug release system containing the above-described aggregates of insulin, the pH-sensitive polymer layer and the insulin aggregates cladding layer sequentially on the substrate may be a pH-sensitive polymer layer, all the insulin aggregated layer combinations, or with a PH sensitive polymer layer, the protein layer covering the aggregate and the insulin sequence layer on the substrate may be a polymer having a PH sensitive layer, a protein layer, insulin All combinations aggregate layer.

[0008] 2、第二种类型:基底为胰岛素聚集体 [0008] 2, the second type: the substrate is insulin aggregates

[0009] 此种载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,由基底及包覆在基底上的具有PH敏感性的聚合物层和蛋白质层组成,基底、具有pH敏感性的聚合物层、蛋白质层相互之间通过静电作用结合成一体;或由基底及包覆在基底上的具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层组成,基底、具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层相互之间通过静电作用结合成一体;所述基底为胰岛素聚集体颗粒。 [0009] Such aggregates containing long acting insulin to control blood glucose concentration of drug release system, the coated substrate and the polymer layer on the substrate and a protein layer having a PH sensitive composition, a substrate having a pH-sensitive polymer layer, between the protein layers are integrally bonded by electrostatic interaction; or a coated substrate and a polymer layer on the substrate having a PH sensitive, protein layer, insulin aggregate layers, the substrate having a PH sensitive of the polymer layer, a protein layer, insulin aggregate layer integrally joined to each other by electrostatic interaction; the substrate is insulin aggregate particle.

[0010] 上述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,具有pH敏感性的聚合物层和蛋白质层在基底上的包覆顺序可以是具有PH敏感性的聚合物层、蛋白质层的所有组合方式,或具有PH敏感性的聚合物层、蛋白质层和胰岛素聚集体层在基底上的包覆顺序可以是具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层的所有组合方式。 [0010] The long-term control of blood glucose concentration of the drug release system containing the above-described aggregates of insulin, the sequence having a coating pH-sensitive polymer layer and a protein layer on the substrate may be a PH sensitive polymer layer, All combinations of the protein layer, or a polymer having a PH sensitive layer, the protein layer and the aggregation of insulin cladding layer sequentially on a substrate may be a PH sensitive polymer layer, a protein layer, a layer of insulin aggregates All combinations.

[0011] 上述两种载有胰岛素聚集体的长效控制血糖浓度的药物控释体系中,所述胰岛素聚集体为牛胰岛素聚集体、重组人胰岛素聚集体、猪胰岛素聚集体中的一种。 [0011] The two types of carrier systems for controlled drug release insulin aggregates of long-term control of blood glucose concentration, the insulin aggregate as a bovine insulin aggregates, aggregates of recombinant human insulin, porcine insulin aggregates. 所述具有pH敏感性的聚合物为聚甲基丙烯酸-N,N-二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯、聚甲基丙烯酸酯、壳聚糖中的一种,分子量为I〜lOOOkda。 The pH-sensitive polymer is polymethacrylic acid -N, N- dimethylaminoethyl acrylate, polymethacrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate, polymethacrylate a chitosan having a molecular weight of I~lOOOkda. 所述蛋白质为葡萄糖氧化酶或过氧化氢酶。 The protein is a glucose oxidase or catalase.

[0012] 本发明所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系的制备方法,有以下两种: [0012] The method of preparing a pharmaceutical insulin aggregates long acting controlled release systems of the present invention, control of blood glucose concentration of the carrier, there are the following two:

[0013] 1、第一种方法 [0013] 1, a first method

[0014] 第一种方法用于制备上述第一种类型的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,工艺步骤如下: Controlled Drug Delivery System [0014] A first method for the preparation of long-term control of blood glucose concentration of the first type contains aggregates of insulin, the following process steps:

[0015] (I)配制原料溶液或悬浊液 [0015] (I) starting material prepared solution or suspension

[0016] 原料溶液或悬浊液为具有pH敏感性的聚合物溶液和胰岛素聚集体悬浊液,或具有pH敏感性的聚合物溶液、胰岛素聚集体悬浊液和蛋白质溶液; [0016] The starting material is a solution or suspension having a pH-sensitive polymer solution and insulin aggregates suspension, or with a pH-sensitive polymer solution, suspension and insulin aggregate protein solution;

[0017] 所述具有pH敏感性的聚合物为聚甲基丙烯酸-N,N- 二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯、聚甲基丙烯酸酯、壳聚糖中的一种,分子量为I〜IOOOkda ;用pH值为I〜7的盐酸水溶液、乙酸溶液或去离子水将具有pH敏感性的聚合物配制成浓度为0.015〜100mg/mL的溶液,若具有pH敏感性的聚合物为聚甲基丙烯酸-N,N-二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯或聚甲基丙烯酸酯,则再加入NaCl,NaCl的加入量以其在混合液中的浓度达到0.01〜5mol/L为限; [0017] The pH-sensitive polymer is polymethacrylic acid -N, N- dimethylaminoethyl acrylate, polymethacrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate, poly A acrylate, a chitosan, a molecular weight of I~IOOOkda; I~7 with the pH of the aqueous solution of hydrochloric acid, acetic acid solution or deionized water having a pH-sensitive polymer formulation at a concentration of 0.015~100mg / mL solution, if the pH-sensitive polymer is polymethacrylic acid -N, N- dimethylaminoethyl acrylate, polymethacrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate or poly methacrylate, was added the NaCl, NaCl is added in an amount such that its concentration in the mixture reaches 0.01~5mol / L is limited;

[0018] 所述胰岛素聚集体为牛胰岛素聚集体、重组人胰岛素聚集体、猪胰岛素聚集体中的一种;用PH值为3〜8的盐酸水溶液、乙酸水溶液、磷酸水溶液或去离子水将胰岛素聚集体配制成浓度为0.015〜100mg/mL的悬池液; [0018] The insulin is bovine insulin aggregates aggregates aggregates of recombinant human insulin, porcine insulin aggregates of one; with a PH value 3~8 aqueous hydrochloric acid, aqueous acetic acid, aqueous phosphoric acid solution or deionized water insulin aggregates were formulated as a liquid suspension cell 0.015~100mg / mL of;

[0019] 所述蛋白质为葡萄糖氧化酶或过氧化氢酶;用pH值为3〜8的盐酸水溶液、乙酸水溶液、磷酸水溶液或去离子水将蛋白质配制成浓度为0.015〜100mg/mL的溶液; [0019] The protein is a glucose oxidase or catalase; 3~8 with aqueous hydrochloric acid a pH, aqueous acetic acid, aqueous phosphoric acid solution or deionized water to a protein concentration of formulated solution 0.015~100mg mL /;

[0020] (2)处理基底表面使其带电荷;[0021] (3)组装重叠层或包覆层 [0020] (2) treating the substrate surface so as charged; [0021] (3) assembling the cladding layer or overlapping layers

[0022] 将具有pH敏感性的聚合物溶液、胰岛素聚集体悬浊液和/或蛋白质溶液用旋涂、浸涂或喷涂的方法按照所述药物控释体系的结构重叠或包覆在基底上和在前的重叠或包覆物上,在制备每一层重叠或包覆物时,静置O-lOOmin,并将形成所述重叠或包覆物的多余溶液或悬浊液用去离子水冲洗、离心或透析的方法去掉并干燥,即得到载有胰岛素聚集体的长效控制血糖浓度的药物控释体系; [0022] The pH-sensitive polymer solution, suspension aggregates of insulin and / or protein solution by spin-coating, dip coating or spray coating methods or overlapping structure according to the drug release system on the substrate and overlapping the front or wrap, in the preparation of each layer overlap or wrap, allowed to stand O-lOOmin, excess solution or suspension is formed and the deionized water of the wrap overlapping or washing, centrifugation or dialysis method of removing and dried to obtain aggregates containing the long-acting insulin to control blood sugar levels controlled drug delivery system;

[0023] 所述基底为平板、微球或医疗器械,所述平板或微球由有机玻璃、石英、云母中的一种制作。 [0023] The substrate is a tablet, microspheres or medical device, the plate or microspheres of glass, quartz, mica method of making.

[0024] 2、第二种方法 [0024] 2, a second method

[0025] 第二种方法用于制备上述第二种类型的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,工艺步骤如下: Controlled Drug Delivery System [0025] The second method for producing the above long-term control of blood glucose concentration of the second type contains aggregates of insulin, the following process steps:

[0026] (I)配制原料溶液或悬浊液 [0026] (I) starting material prepared solution or suspension

[0027] 原料溶液或悬浊液为具有pH敏感性的聚合物溶液和蛋白质溶液,或具有pH敏感性的聚合物溶液、蛋白质溶液和胰岛素聚集体悬浊液; [0027] The starting material is a solution or suspension having a pH-sensitive polymer solution and a protein solution, or a pH-sensitive polymer solution, the protein solution and the suspension of insulin aggregates;

[0028] 所述具有pH敏感性的聚合物为聚甲基丙烯酸-N,N- 二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯、聚甲基丙烯酸酯、壳聚糖中的一种,分子量为I〜IOOOkda ;用pH值为I〜7的盐酸水溶液、乙酸溶液或去离子水将具有pH敏感性的聚合物配制成浓度为0.015〜100mg/mL的溶液,若具有pH敏感性的聚合物为聚甲基丙烯酸-N,N-二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯或聚甲基丙烯酸酯,则再加入NaCl,NaCl的加入量以其在混合液中的浓度达到0.01〜5mol/L为限; [0028] The pH-sensitive polymer is polymethacrylic acid -N, N- dimethylaminoethyl acrylate, polymethacrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate, poly A acrylate, a chitosan, a molecular weight of I~IOOOkda; I~7 with the pH of the aqueous solution of hydrochloric acid, acetic acid solution or deionized water having a pH-sensitive polymer formulation at a concentration of 0.015~100mg / mL solution, if the pH-sensitive polymer is polymethacrylic acid -N, N- dimethylaminoethyl acrylate, polymethacrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate or poly methacrylate, was added the NaCl, NaCl is added in an amount such that its concentration in the mixture reaches 0.01~5mol / L is limited;

[0029] 所述胰岛素聚集体为牛胰岛素聚集体、重组人胰岛素聚集体、猪胰岛素聚集体中的一种;用PH值为3〜8的盐酸水溶液、乙酸水溶液、磷酸水溶液或去离子水将胰岛素聚集体配制成浓度为0.015〜100mg/mL的悬池液; [0029] The insulin is bovine insulin aggregates aggregates aggregates of recombinant human insulin, porcine insulin aggregates of one; with a PH value 3~8 aqueous hydrochloric acid, aqueous acetic acid, aqueous phosphoric acid solution or deionized water insulin aggregates were formulated as a liquid suspension cell 0.015~100mg / mL of;

[0030] 所述蛋白质为葡萄糖氧化酶或过氧化氢酶;用pH值为3〜8的盐酸水溶液、乙酸水溶液、磷酸水溶液或去离子水将蛋白质配制成浓度为0.015〜100mg/mL的溶液; [0030] The protein is a glucose oxidase or catalase; 3~8 with aqueous hydrochloric acid a pH, aqueous acetic acid, aqueous phosphoric acid solution or deionized water to a protein concentration of formulated solution 0.015~100mg mL /;

[0031] (2)制备胰岛素聚集体基底 [0031] (2) Preparation of insulin aggregates substrate

[0032] 将胰岛素聚集体用pH 2〜5的乙酸或盐酸溶液配制成浓度为I〜100mg/mL悬浊液,然后用NaOH溶液调节聚集体悬浊液的pH值至6〜10,然后加入NaCl,NaCl的加入量以混合液中NaCl的浓度达到0.5〜5mol/L,在室温、常压下搅拌混合I〜20h,继后离心分离,获带负电荷的颗粒状胰岛素聚集体基底; [0032] The aggregates of insulin formulated with acetic acid or hydrochloric acid solution pH to a concentration of 2 ~ 5 I~100mg / mL suspension, and then adjust the aggregate suspension to pH 6~10 with NaOH solution, followed by addition of NaCl, NaCl is added in an amount to achieve a concentration of NaCl mixture 0.5~5mol / L, stirring and mixing at room temperature, atmospheric pressure I~20h, following centrifugation, is eligible for a negatively charged insulin aggregate particulate substrate;

[0033] (3)组装包覆层 [0033] (3) assembling the cladding layer

[0034] 将具有pH敏感性的聚合物溶液和蛋白质溶液采用与基底混合和与在前的包覆物混合的方法按照所述药物控释体系的结构包覆在基底上和在前的包覆物上,或将具有PH敏感性的聚合物溶液、蛋白质溶液和胰岛素聚集体悬浊液采用与基底混合和与在前的包覆物混合的方法按照所述药物控释体系的结构包覆在基底上和在前的包覆物上,在制备每一层包覆物时,搅拌至少lOmin,并将形成所述包覆物的多余溶液或悬浊液用去离子水冲洗、离心或透析的方法去掉,即得到载有胰岛素聚集体的长效控制血糖浓度的药物控释体系。 [0034] The pH-sensitive polymer solution and protein solution using the method of mixing the base and mixing with previous wrap and coated on a substrate coated front structure according to the drug release system on the object, or a polymer solution having a PH sensitive protein solution and using a suspension of insulin aggregates and mixing method of mixing the substrate with the previous wrap of the coated structure according to the drug release system the upper substrate and the front coating was coated in the preparation of each layer was stirred for lOmin at least, is formed and the excess coating solution or suspension was rinsed with deionized water, centrifugation or dialysis removing method, to obtain aggregates containing the long-acting insulin to control blood glucose concentration of drug release system.

[0035] 本发明所述方法中,牛胰岛素聚集体、重组人胰岛素聚集体的制备方法见下述文献:[0036] IΛGupta, S.,Chattopadhyay, T.,Singh, MP,et al.,Supramolecular insulinassembly II for a sustained treatment of type I diabetes mellitus.Proc.Natl.Acad.Sc1.USA 2010,107:13246-13251.[0037] 本发明所述方法中,猪胰岛素聚集体的制备方法如下: [0035] The method of the invention, bovine insulin preparation see aggregates, aggregates of recombinant human insulin following documents: [0036] IΛGupta, S., Chattopadhyay, T., Singh, MP, et al, Supramolecular. insulinassembly II for a sustained treatment of type I diabetes mellitus.Proc.Natl.Acad.Sc1.USA 2010,107:. 13246-13251 [0037] the method of the present invention, porcine insulin aggregates prepared as follows:

[0038] 将猪胰岛素用溶液配制成浓度在0.01mg/mL-400mg/mL之间的溶液,将该溶液置于反应容器中在搅拌下于常压、5〜60°C反应I〜300h,反应时间届满后,得猪胰岛素聚集体悬浊液,然后将猪胰岛素聚集体悬浊液冷冻干燥即得到猪胰岛素聚集体;所述溶液为盐酸水溶液,或醋酸水溶液,或盐酸和醋酸的混合水溶液,或为磷酸盐缓冲液,或为柠檬酸缓冲液,或为氢氧化钠水溶液,它们的pH值在I〜14之间。 [0038] The porcine insulin was formulated with a concentration of the solution between 0.01mg / mL-400mg / mL, and the solution was placed in a reaction vessel with stirring under a normal pressure, 5~60 ° C in the reaction I~300h, after the expiration of the reaction time, to obtain a suspension of aggregates porcine insulin, porcine insulin was then freeze-dried suspension aggregates i.e. porcine insulin aggregates obtained; the mixed aqueous solution is an aqueous hydrochloric acid or aqueous acetic acid, hydrochloric acid and acetic acid, or or phosphate buffer, or citrate buffer solution, or aqueous sodium hydroxide solution, the pH thereof between I~14.

[0039] 本发明具有以下有益效果: [0039] The present invention has the following advantages:

[0040] 1、试验表明,将本发明所述药物控释体系埋植于糖尿病模型动物皮下,可以控制其体内正常血糖浓度达295天(见实施例7),与现有技术相比,大大延长了单次用药控制体内正常血糖浓度的时间。 [0040] 1, tests showed that the drug release system of the present invention is subcutaneously implanted in an animal model of diabetes, which can be controlled euglycemia in vivo concentration of 295 days (see Example 7), compared to the prior art, greatly extending the control normal blood glucose concentration in vivo single dose time.

[0041] 2、本发明所述药物控释体系具有葡萄糖浓度响应性能,是一种智能响应性的胰岛素控制释放体系。 [0041] 2, the present invention is a pharmaceutical controlled release system having glucose concentration response performance, is an intelligent control responsive insulin delivery system.

[0042] 3、本发明所述药物控释体系中的聚合物和蛋白质均为无毒性、生物相容性好的物质,因而其生物相容性好,对机体无毒副作用。 [0042] 3, the present invention is a pharmaceutical controlled release systems of polymers and proteins are non-toxic, biocompatible material good, and therefore good biocompatibility, toxic side effects on the body.

[0043] 4、本发明所述方法操作简单,便于工业化生产。 [0043] 4, the method of the present invention is simple, easy industrial production.

附图说明 BRIEF DESCRIPTION

[0044]图1是本发明所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系的一种结构示意图,图中,1:基底;2:具有pH敏感性的聚合物层;3:胰岛素聚集体层;4:葡萄糖氧化酶层;5:过氧化氢酶层。 [0044] FIG. 1 is the carrier of the present invention there is a system schematic view of long-term drug release control of blood glucose concentrations of insulin aggregates, the drawings, 1: a substrate; 2: pH-sensitive polymer layer; 3: insulin aggregate layer; 4: glucose oxidase layer; 5: catalase layer.

[0045]图2是实施例1所制备的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系在葡萄糖溶液中智能响应性的激光扫描共聚焦显微镜(CLSM)图片,其中,图2(a)为在浓度Og/L葡萄糖溶液中智能关闭的激光扫描共聚焦显微镜(CLSM)图片,图2(b)为在浓度5g/L葡萄糖溶液中智能打开的激光扫描共聚焦显微镜(CLSM)图片。 [0045] FIG. 2 is a long-term control of blood glucose concentration of the drug release system containing insulin aggregates prepared in Example 1 in the glucose solution responsive smart laser scanning confocal microscope (the CLSM) images, wherein FIG. 2 (a) is in a concentration Og / L glucose solution smart off laser scanning confocal microscope (the CLSM) images, FIG. 2 (b) is at a concentration of 5g / L glucose solution smart open laser scanning confocal microscope (the CLSM) image.

[0046] 图3是将本发明所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系埋植在糖尿病模型动物体内,其体内血糖浓度的变化曲线。 [0046] FIG. 3 is a carrier of the present invention, the aggregate long-acting insulin to control blood glucose concentration of drug release system implanted in vivo animal models of diabetes, blood glucose concentration in the body changes its curve.

具体实施方式 detailed description

[0047] 下面通过实施例对本发明所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系及其制备方法和应用作进一步说明。 [0047] The following examples contain insulin aggregates of the present invention, long-term control of blood glucose concentration of the drug release system and a preparation method and application further illustrated.

[0048] 实施例1 [0048] Example 1

[0049] 本实施例的工艺步骤如下: [0049] The process steps of the present embodiment is as follows:

[0050] (I)制备猪胰岛素聚集体 [0050] (I) Preparation of porcine insulin aggregates

[0051] 将猪胰岛素用pH值为6.5的磷酸盐缓冲溶液配制成浓度为10mg/mL的溶液,将该溶液置于反应容器中在搅拌下于常压、35°C反应26h,反应时间届满后,得猪胰岛素聚集体悬浊液,然后将猪胰岛素聚集体悬浊液在_80°C冷冻干燥15h即得到猪胰岛素聚集体(在下述描述中简称P-SIA); [0051] The porcine insulin with a pH 6.5 phosphate buffer solution formulated at a concentration of 10mg / mL of solution which was placed in a reaction vessel with stirring at atmospheric pressure, 35 ° C 26h is the reaction, the expiry of the reaction time after give aggregates suspension porcine insulin, porcine insulin and the aggregate suspension at _80 ° C to obtain freeze-dried 15h porcine insulin aggregates (referred to in the following description P-SIA);

[0052] (2)配制原料溶液或悬浊液 [0052] (2) Preparation of raw material solution or suspension

[0053] 将分子量为120kda的星型聚合物:聚甲基丙烯酸-N,N-二甲氨基乙酯(在下述描述中简称Star-PDMAEMA)用pH值为5的盐酸水溶液配制成浓度为0.5mg/mL的溶液,然后加入NaCl,NaCl的加入量以其在混合液中的浓度达到0.2mol/L为限; [0053] The molecular weight of the star polymer 120kda: aqueous hydrochloric acid polymethacrylic acid -N, N- dimethylaminoethyl methacrylate (referred to in the following description Star-PDMAEMA) formulated with a pH of 5 at a concentration of 0.5 concentration mg / mL solution of NaCl was then added, the addition amount of NaCl in the mixture reaches its 0.2mol / L is limited;

[0054] 将步骤(I)制备的P-SIA用去离子水配制成浓度为2mg/mL的P-SIA悬浊液; [0054] P-SIA prepared in step (I) with deionized water to a concentration of 2mg / mL suspension of the P-SIA;

[0055] 将葡萄糖氧化酶(在下述描述中简称G0D)粉末用pH值为6的盐酸水溶液配制成浓度为3mg/mL的GOD溶液;将过氧化氢酶(在下述描述中简称CAT)粉末用去离子水配制成浓度为2mg/mL的CAT溶液; [0055] The aqueous hydrochloric acid solution of glucose oxidase (referred to in the following description G0d) powder with a pH of 6, were formulated as a solution GOD 3mg / mL; and the catalase (referred to in the following description CAT) powder deionized water were formulated as CAT solution 2mg / mL of;

[0056] (3)处理基底 [0056] (3) treating the substrate

[0057] 基底为有机玻璃片,其长度为2cm、宽度为2cm、厚度为2mm。 [0057] The substrate is a PMMA sheet having a length of 2cm, width of 2cm, thickness 2mm. 首先用去离子水清洗3次,然后在体积比为1:1的去离子水与异丙醇的混合液中浸泡15分钟,再放入体积比5:1:1的去离子水/质量浓度30% H2O2/质量浓度29%氨水的混合溶液中,在70°C水浴条件下浸泡10分钟,继后用去离子水冲洗后干燥,即获得表面带负电荷的有机玻璃片。 First, washed three times with deionized water, and then in a volume ratio of 1: a mixture of isopropanol and deionized water in a soak for 15 minutes, then add a volume ratio of 5: 1: 1 deionized water / concentration 30% H2O2 / 29% by mass concentration of a mixed solution of ammonia water, for 10 minutes at 70 ° C for cooling, and the following rinsing with deionized water and dried to obtain a PMMA sheet surface i.e. negatively charged.

[0058] (4)组装覆盖层 [0058] (4) assembling the cover layer

[0059] 本实施例制备的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系中,Star-PDMAEMA层、P-SIA层、GOD层、CAT层在基底上的重叠顺序为:{(Star-PDMAEMA/P-SIA) 2+ (Star-PDMAEMA/CAT)!+ (Star-PDMAEMA/GOD) J 2+Star-PDMAEMA,其结构见图1。 Long-term control of blood glucose concentration of the drug release system containing insulin aggregates [0059] Preparation Example of the present embodiment, Star-PDMAEMA layer, P-SIA layer, a layer of GOD, overlapping order CAT layer on the substrate is: { (Star-PDMAEMA / P-SIA) 2+ (Star-PDMAEMA / CAT)! + (Star-PDMAEMA / GOD) J 2 + Star-PDMAEMA, the structure shown in Figure 1.

[0060] 按照上述结构,首先在室温、常压下将步骤⑵配制的Star-PDMAEMA溶液150 μ L涂覆于经过表面处理的有机玻璃片表面,吸附30min,然后用去离子水清洗表面,并用氮气吹干;再将步骤⑵配制的P-SIA悬浊液200yL涂覆于Star-PDMAEMA层表面,吸附40min,然后用去离子水清洗表面,并用氮气吹干;重复上述操作,得到2个Star-PDMAEMA/P-SI双分子层。 [0060] According to the above configuration, Star-PDMAEMA solution of 150 μ L is first applied at room temperature, the pressure in the step of formulating ⑵ PMMA sheet surface after the surface treatment, adsorption 30min, and then surface washed with deionized water, and dried dry nitrogen; ⑵ step then formulated P-SIA suspension 200yL Star-PDMAEMA applied to the surface layer, the adsorption 40min, and then surface washed with deionized water, and blown dry with nitrogen; above operation is repeated to give 2 Star -PDMAEMA / P-SI bilayer. 在室温、常压下将步骤⑵配制的Star-PDMAEMA 150yL溶液涂覆于P-SIA层表面,吸附30min,然后用去离子水清洗表面,并用氮气吹干,再将步骤(2)配制的CAT溶液150 μ L涂覆于Star-PDMAEMA层表面,吸附40min,然后用去离子水清洗表面,并用氮气吹干,得到I个Star-PDMAEMA/CAT双分子层。 Star-PDMAEMA 150yL solution was coated at room temperature and atmospheric pressure in the step of formulating ⑵ P-SIA surface layer, the adsorption 30min, and then surface washed with deionized water, and blown dry with nitrogen, then Step (2) Preparation of CAT solution of 150 μ L Star-PDMAEMA applied to the surface layer, the adsorption 40min, and then surface washed with deionized water, and blown dry with nitrogen to afford the I Star-PDMAEMA / CAT bilayer. 在室温、常压下将步骤⑵配制的Star-PDMAEMA溶液150 μ L涂覆于CAT层表面,吸附30min,然后用去离子水清洗表面,并用氮气吹干,再将步骤(2)配制的GOD溶液200 μ L涂覆于Star-PDMAEMA层表面,吸附40min,然后用去离子水清洗表面,并用氮气吹干;重复上述操作,得到2个Star-PDMAEMA/GOD双分子层。 At room temperature, and pressure of step ⑵ formulated Star-PDMAEMA was applied to the surface of 150 μ L CAT layer, adsorption 30min, and then surface washed with deionized water, and blown dry with nitrogen, then Step (2) Preparation of GOD solution of 200 μ L Star-PDMAEMA applied to the surface layer, the adsorption 40min, and then surface washed with deionized water, and blown dry with nitrogen; above operation is repeated to give 2 Star-PDMAEMA / GOD bilayer. 至此,在基底上形成了下述覆盖顺序的“(Star-PDMAEMA/P-SIA)2+(Star-PDMAEMA/CAT) J(Star-PDMAEMAAiOD)2”的10层覆盖层。 Thus, on the substrate is formed "(Star-PDMAEMA / P-SIA) 2+ (Star-PDMAEMA / CAT) J (Star-PDMAEMAAiOD) 2" 10 of the following layer covers the covering layer in order. 重复上述操作,得到基底上形成了下述覆盖顺序的“ {(Star-PDMAEMA/P-SIA) 2+ (Star-PDMAEMA/CAT)汴(Star-PDMAEMA/GOD) 2} 2 ”的20 层覆盖层。 Repeating the above operation, the obtained formed "{(Star-PDMAEMA / P-SIA) 2+ (Star-PDMAEMA / CAT) Bian (Star-PDMAEMA / GOD) 2} 2" 20 in order to cover the following layer covers the substrate Floor. 最后,在室温、常压下将150 μ L Star-PDMAEMA溶液涂覆于GOD层上,然后用去离子水清洗表面,并用氮气吹干,即得到图1所述结构的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系。 Finally, at room temperature, normal pressure Star-PDMAEMA solution was coated on a 150 μ L GOD layer and then surface washed with deionized water, and blown dry with nitrogen, to obtain a structure of the carrier of FIG insulin aggregates long-term control of blood glucose concentration of drug release system.

[0061] 在室温、常压下,将本实施例所制备的载有胰岛素聚集体的药物控释体系分别放入浓度Og/L葡萄糖溶液中和5g/L的葡萄糖溶液中,其激光扫描共聚焦显微镜(CLSM)图片见图2,从图2可以看出,在Og/L葡萄糖溶液中本体系处于关闭状态,而在5g/L的葡萄糖溶液中本体系处于开放状态。 Drug release system of insulin aggregates containing [0061] at room temperature and atmospheric pressure, the Production Example of the present embodiment are placed in the concentration Og / L glucose solution and 5g / L glucose solution, which is a laser scanning confocal confocal microscopy (the CLSM) image shown in Figure 2, can be seen from Figure 2, in Og / L glucose solution in the present system is turned off, while 5g / L glucose solution in the present system in an open state. [0062] 实施例2 [0062] Example 2

[0063] 本实施例的工艺步骤如下: [0063] The process steps of the present embodiment is as follows:

[0064] (I)制备牛胰岛素聚集体 [0064] (I) Preparation of bovine insulin aggregates

[0065] 将牛胰岛素用pH值为5.9的盐酸水溶液配制成浓度为0.01mg/mL的溶液,将该溶液置于反应容器中在搅拌下于常压、25°C反应43h,反应时间届满后,得牛胰岛素聚集体悬浊液,然后将牛胰岛素聚集体悬浊液在_60°C冷冻干燥18h即得到牛胰岛素聚集体(在下述描述中简称B-SIA); [0065] with bovine insulin a pH 5.9 aqueous hydrochloric acid formulated at a concentration of 0.01mg / mL was added and the solution was placed in a reaction vessel with stirring under a normal pressure, 25 ° C 43h the reaction, the expiry of the reaction time to obtain a suspension of aggregates of bovine insulin, bovine insulin and the aggregate suspension at _60 ° C to obtain freeze-dried 18h ​​bovine insulin aggregates (referred to as B-SIA in the following description);

[0066] (2)配制原料溶液 [0066] (2) Preparation of raw material solution

[0067] 将分子量为170kda的星型聚合物:聚甲基丙烯酸_N,N-二甲氨基乙酯(在下述描述中简称Star-PDMAEMA)用pH值为6.2的盐酸水溶液配制成浓度为10mg/mL的溶液,然后加入NaCl,NaCl的加入量以其在混合液中的浓度达到0.lmol/L为限; [0067] The molecular weight of the star polymer 170kda: polymethacrylates _N, N- dimethylaminoethyl methacrylate (referred to in the following description Star-PDMAEMA) is formulated in an aqueous solution of 6.2 pH hydrochloric acid with a concentration of 10mg / mL was then added NaCl, NaCl is added in an amount such that its concentration in the mixture reaches 0.lmol / L is limited;

[0068] 将葡萄糖氧化酶(在下述描述中简称GOD)粉末用pH值为6的盐酸水溶液配制成浓度为10mg/mL的GOD溶液;将过氧化氢酶(在下述描述中简称CAT)粉末用pH值为6的盐酸水溶液配制成浓度为10mg/mL的CAT溶液; [0068] The aqueous hydrochloric acid solution of glucose oxidase (GOD referred to in the following description) powder with a pH of 6 is formulated to a concentration of GOD solution 10mg / mL; and the catalase (referred to in the following description CAT) powder pH of aqueous hydrochloric acid at a concentration of 6 formulated 10mg / mL solution of the CAT;

[0069] (3)制备胰岛素聚集体基底 [0069] (3) insulin aggregates prepared substrate

[0070] 将步骤(I)制备的B-SIA用pH值为3的盐酸溶液配制成浓度为100mg/mL的B-SIA悬浊液,然后用浓度lmol/L的NaOH溶液调节B-SIA悬浊液的pH值至6 (牛胰岛素等电点以上),再加入NaCl,NaCl的加入量以混合液中NaCl的浓度达到0.5mol/L为限,在室温、常压下搅拌混合lh,继后以1000转/分的转速离心5min,获带负电荷的颗粒状B-SIA基底。 [0070] The B-SIA prepared in step (I) with pH 3 hydrochloric acid solution formulated at a concentration of 100mg / mL suspension of B-SIA, and then adjusted with a concentration of lmol / L NaOH solution B-SIA suspension turbid solution to pH 6 (above the isoelectric point of bovine insulin), NaCl was added, the amount of NaCl was added at a concentration of NaCl in the mixture reaches 0.5mol / L limited to, stirring and mixing at room temperature for lh, atmospheric pressure, following the after at 1000 r / min centrifuged 5min, eligible negatively charged particulate substrate B-SIA.

[0071] (4)组装包覆层 [0071] (4) assembling the cladding layer

[0072] 本实施例制备的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系中,Star-PDMAEMA层、GOD层、CAT层在颗粒状B-SIA基底上的包覆顺序为:(Star-PDMAEMA/GOD) 2+ (Star-PDMAEMA/CAT) J (Star-PDMAEMA/GOD) 2+Star_PDMAEMA。 Long-term control of blood glucose concentration of the drug release system containing insulin aggregates [0072] Preparation Example of the present embodiment, Star-PDMAEMA layer, GOD layer, CAT layers on the particulate coated substrate B-SIA order: (Star-PDMAEMA / GOD) 2+ (Star-PDMAEMA / CAT) J (Star-PDMAEMA / GOD) 2 + Star_PDMAEMA.

[0073] 按照上述结构,在室温、常压下将步骤(3)制备的颗粒状B-SIA基底20mg与步骤(2)配制的Star-PDMAEMA溶液5mL通过搅拌的方式混合,搅拌时间20min,搅拌时间届满后,以1000转/分的转速离心5min,然后将所获沉淀用浓度ImM的HCl与浓度0.5M的NaCl混合液洗涤、离心2次,获包覆有Star-PDMAEMA层的微粒;在室温、常压下将包覆有Star-PDMAEMA层的微粒与步骤(2)配制的GOD溶液3mL通过搅拌的方式混合,搅拌时间20min,搅拌时间届满后,以1000转/分的转速离心5min,然后将所获沉淀用浓度ImM的HCl与浓度0.5M的NaCl混合液洗涤、离心2次,获依次包覆有Star_PDMAEMA、GOD层的微粒,即I个Star-PDMAEMA/GOD双分子层。 [0073] According to the above configuration, Star-PDMAEMA 5mL solution at room temperature, the pressure in step (3) B-SIA particulate substrate prepared in 20mg and step (2) is prepared by way of mixing with stirring, mixing time 20min, stirring after the time expires, at 1000 revolutions / minute centrifuged 5min, then the mixture was washed with NaCl concentrations of 0.5M HCl and the precipitate obtained with concentrations of ImM, centrifugation twice, was coated with a fine layer of Star-PDMAEMA; in at room temperature, covered with fine particles at atmospheric pressure and a step Star-PDMAEMA layer (2) GOD solution was prepared by mixing stirring 3mL way, mixing time 20min, stirring time after the expiration of, at 1000 rev / min centrifuged 5min, the resulting mixture was then precipitated with 0.5M NaCl HCl concentration and the concentration of ImM washed twice by centrifugation, was sequentially coated with Star_PDMAEMA, microparticles GOD layer, i.e., the I Star-PDMAEMA / GOD bilayer. 重复上述操作,得到2个Star-PDMAEMA/GOD双分子层。 Above operation is repeated to give 2 Star-PDMAEMA / GOD bilayer.

[0074] 在室温、常压下将依次包覆有Star-PDMAEMA、GOD层、Star-PDMAEMA、GOD层的微粒与步骤(2)配制的Star-PDMAEMA溶液5mL通过搅拌的方式混合,搅拌时间20min,搅拌时间届满后,以1000转/分的转速离心5min,然后将所获沉淀用浓度ImM的HCl与浓度 [0074] The coated successively at room temperature and atmospheric pressure with a Star-PDMAEMA, GOD layer, Star-PDMAEMA, fine particles and the step of GOD layer (2) Star-PDMAEMA prepared by mixing a solution of 5mL of stirring manner, mixing time 20min after stirring time expires at 1000 r / min centrifuged 5min, then the precipitate was obtained by concentration of HCl with a concentration of ImM

0.5M的NaCl混合液洗涤、离心2次;在室温、常压下将前述微粒与步骤(2)配制的CAT溶液ImL通过搅拌的方式混合,搅拌时间20min,搅拌时间届满后,以1000转/分的转速离心5min,然后将所获沉淀用浓度ImM的HCl与浓度0.5M的NaCl混合液洗涤、离心2次,获I个Star-PDMAEMA/CAT双分子层。 The mixture was washed with 0.5M NaCl, and centrifuged for 2 times; ImL the CAT solution and the fine particles in step (2) is prepared by mixing at room temperature and stirred manner, pressure, mixing time 20min, stirring time after the expiration of, at 1000 revolutions / min centrifuged 5min, then the resulting precipitate mixture was washed with 0.5M NaCl HCl concentration and the concentration of ImM, centrifuging twice the I eligible Star-PDMAEMA / CAT bilayer. 至此,在基底上形成了下述包覆顺序的“(Star-PDMAEMA/GOD) 2+ (Star-PDMAEMA/CAT)! ”的6层包覆层。 Thus, the following are formed on the substrate coated sequence "(Star-PDMAEMA / GOD) 2+ (Star-PDMAEMA / CAT)!" Cladding layer 6. 重复上述操作,在基底上形成了下述包覆顺序的“(Star-PDMAEMA/GOD) J(Star-PDMAEMAZCAT)1+(Star-PDMAEMA/GOD)2” 的IO 层包覆层。 Repeating the above operations, it is formed on the substrate coated with the following sequence "(Star-PDMAEMA / GOD) J (Star-PDMAEMAZCAT) 1+ (Star-PDMAEMA / GOD) 2" IO layer cladding layer. 最后,在室温、常压下上述10层包覆层的微粒与步骤(2)配制的Star-PDMAEMA溶液2mL通过搅拌的方式混合,搅拌时间20min,搅拌时间届满后,以1000转/分的转速离心5min,然后将所获沉淀用浓度ImM的HCl与浓度0.5M的NaCl混合液洗涤、离心2次,即得到本实施例所述结构的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,将所制得的微球状药物控释体系在-60°C冻干干燥17h。 Finally, the above-described fine particles and the step of cladding layer 10 (2) Star-PDMAEMA formulated 2mL of mixed at room temperature, normal pressure by way of stirring, mixing time 20min, stirring time after the expiration of, at 1000 rev / min speed centrifuge 5min, then washed with the resulting concentration of ImM HCl concentration of 0.5M NaCl and precipitated mixture was centrifuged twice, i.e., to obtain the present configuration of the carrier according to a long-acting insulin to control blood glucose aggregate concentration of drug control embodiment release system, the prepared microspheres the controlled release medicament system at -60 ° C freeze-dried 17h.

[0075] 实施例3 [0075] Example 3

[0076] 本实施例的工艺步骤如下: [0076] The process steps of the present embodiment is as follows:

[0077] (I)制备重组人胰岛素聚集体 [0077] (I) Preparation of recombinant human insulin aggregates

[0078] 将重组人胰岛素用pH值为4.9的醋酸水溶液配制成浓度为400mg/mL的溶液,将该溶液置于反应容器中在搅拌下于常压、5°C反应48h,反应时间届满后,得重组人胰岛素聚集体悬浊液,然后将重组人胰岛素聚集体悬浊液在_80°C冷冻干燥9h即得到重组人胰岛素聚集体(在下述描述中简称rH-SIA); [0078] The use of recombinant human insulin a pH 4.9 aqueous acetic acid is formulated as a solution at a concentration of 400mg mL /, the solution was placed in a reaction vessel with stirring at atmospheric pressure, 5 ° C the reaction 48h, after expiry of the reaction time to obtain a suspension of aggregates of recombinant human insulin, recombinant human insulin and the aggregate suspension at _80 ° C to obtain freeze-dried recombinant human insulin 9h aggregates (referred to in the following description rH-SIA);

[0079] (2)配制原料溶液 [0079] (2) Preparation of raw material solution

[0080] 将分子量为IOkda的壳聚糖用pH值为6.2的乙酸水溶液配制成浓度为lmg/mL的壳聚糖溶液;将葡萄糖氧化酶(在下述描述中简称G0D)粉末用pH值为5的盐酸水溶液配制成浓度为3mg/mL的GOD溶液。 [0080] The molecular weight of chitosan IOkda with a pH 6.2 aqueous acetic acid formulated at a concentration of lmg / mL chitosan solution; glucose oxidase (referred to in the following description G0d) powder with a pH of 5 formulated aqueous hydrochloric acid solution to a concentration of GOD 3mg / mL of.

[0081] (3)制备胰岛素聚集体基底 [0081] (3) insulin aggregates prepared substrate

[0082] 将步骤(I)制备的rH-SIA用pH值为5的盐酸溶液配制成浓度为lmg/mL的rH_SIA悬浊液,然后用浓度0.1N的NaOH溶液调节rH-SIA悬浊液的pH值至10,然后加入NaCl,NaCl的加入量以混合液中NaCl的浓度达到5mol/L为限,在室温、常压下搅拌混合20h,继后以3000转/分的转速离心15min,获带负电荷的颗粒状rH-SIA基底。 [0082] rH-SIA prepared in step (I) with a solution of hydrochloric acid a pH of 5 is formulated at a concentration of lmg / mL in rH_SIA suspension, then the suspension is adjusted rH-SIA concentration of 0.1N NaOH solution to pH 10, followed by addition of NaCl, the amount of NaCl was added at a concentration of NaCl in the mixture reaches 5mol / L is limited, stirred at room temperature for 20 h mixing pressure, followed by the 3000 r / min centrifuged 15min, eligible negatively charged particulate rH-SIA substrate.

[0083] (4)组装包覆层 [0083] (4) assembling the cladding layer

[0084] 本实施例制备的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系中,壳聚糖层、GOD层在颗粒状rH-SIA基底上的包覆顺序为:(壳聚糖/G0D) 1(|。 Long-term control of blood glucose concentration of the drug release system containing insulin aggregates [0084] Preparation Example of the present embodiment, the chitosan layer, cladding layer sequentially GOD rH-SIA on the particulate substrate is chitosan :( sugar / G0D) 1 (|.

[0085] 按照上述结构,在室温、常压下将步骤(3)制备的颗粒状rH-SIA基底IOmg与步骤(2)配制的壳聚糖溶液5mL通过搅拌的方式混合,搅拌时间60min,搅拌时间届满后,以3000转/分的转速离心5min,然后将所获沉淀用浓度2mM的HCl与浓度0.7M的NaCl混合液洗涤、离心2次,获包覆有壳聚糖层的微粒;在室温、常压下将包覆有壳聚糖层的微粒与步骤(2)配制的GOD溶液ImL通过搅拌的方式混合,搅拌时间IOmin,搅拌时间届满后,以3000转/分的转速离心5min,然后将所获沉淀用浓度2mM的HCl与浓度0.7M的NaCl混合液洗涤、离心2次,获依次包覆有壳聚糖层、GOD层的微粒,即I个壳聚糖/GOD双分子层。 [0085] According to the above structure, at room temperature, the pressure in step (3) a particulate rH-SIA IOmg substrate prepared in Step (2) 5mL chitosan solution prepared by way of mixing with stirring, mixing time 60min, stirring after the time expires, at 3000 rev / min centrifuged 5min, then the resulting precipitate was washed with a mixture of NaCl with a concentration of 2mM HCl to a concentration of 0.7M, centrifuged twice coated fine particles obtained chitosan layer; in at room temperature, at atmospheric pressure and a step of chitosan particles coated with layer (2) GOD solution was prepared by mixing ImL stirring way and the stirring time IOmin, stirring time after the expiration, centrifuged at 3000 rev / min 5min, the concentration obtained was then washed with 0.7M concentration of 2mM HCl with a mixture of NaCl precipitation, centrifugation twice, was sequentially coated with a layer of chitosan, GOD fine particle layer, i.e., the I chitosan / GOD bilayer . 重复上述操作,在基底上形成了下述包覆顺序的“(壳聚糖/G0D)1(l”的20层包覆层,即本实施例所述结构的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,将所制得的微球状药物控释体系在_80°C冻干干燥9h。 Repeating the above operations, it is formed on the substrate coated with the following sequence "(chitosan / G0D) 1 (l" cladding layer 20, i.e., the carrier of the present embodiment has a structure of a long-acting insulin aggregates control of blood glucose concentration of the drug release system, the prepared microspheres the controlled release medicament system at _80 ° C freeze-dried 9h.

[0086] 实施例4 [0086] Example 4

[0087] 本实施例的工艺步骤如下: [0087] The process steps of the present embodiment is as follows:

[0088] (I)制备重组人胰岛素聚集体 [0088] (I) Preparation of recombinant human insulin aggregates

[0089] 将重组人胰岛素用pH值为6的柠檬酸水溶液配制成浓度为200mg/mL的溶液,将该溶液置于反应容器中在搅拌下于常压、30°C反应30h,反应时间届满后,得重组人胰岛素聚集体悬浊液,然后将重组人胰岛素聚集体悬浊液在-70°C冷冻干燥12h即得到重组人胰岛素聚集体(在下述描述中简称rH-SIA); [0089] Recombinant human insulin with aqueous citric acid pH of 6 is formulated at a concentration of 200mg / mL of solution which was placed in a reaction vessel with stirring at atmospheric pressure, 30 ° C 30h the reaction, the expiry of the reaction time after give aggregates of recombinant human insulin suspension, human insulin and the aggregate suspension at -70 ° C freeze-dried to obtain 12h recombinant human insulin aggregates (referred to in the following description rH-SIA);

[0090] (2)配制原料溶液 [0090] (2) Preparation of raw material solution

[0091] 将分子量为Ikda的线型聚合物:聚丙烯酸二甲氨基乙酯(在下述描述中简称PDMAEA)用pH值为5的盐酸水溶液配制成浓度为0.5mg/mL的溶液,然后加入NaCl,NaCl的加入量以其在混合液中的浓度达到0.6mol/L为限; [0091] The molecular weight of the linear polymer Ikda: polyacrylic acid, dimethylaminoethyl methacrylate (referred to in the following description PDMAEA) aqueous hydrochloric acid with a pH of 5 was formulated at a concentration of 0.5mg / mL was then added NaCl , NaCl is added in an amount such that its concentration in the mixture reaches 0.6mol / L is limited;

[0092] 将将步骤(I)制备的rH-SIA用去离子水配制成浓度为lmg/mL的rH_SIA悬浊液; [0092] The rH-SIA prepared in the step (I) with deionized water to a concentration of lmg / mL suspension of rH_SIA;

[0093] 将葡萄糖氧化酶(在下述描述中简称G0D)粉末用pH值为6的盐酸水溶液配制成浓度为2mg/mL的GOD溶液; [0093] Glucose oxidase (referred to in the following description G0d) aqueous hydrochloric acid with a pH value of 6 powders were formulated as GOD solution 2mg / mL of;

[0094] (3)处理基底 [0094] (3) treating the substrate

[0095] 基底为云母片,其长度为2cm、宽度为2cm、厚度为2mm,处理方法与实施例1相同。 [0095] The substrate is a mica sheet, a length of 2cm, width of 2cm, a thickness of 2mm, the same processing method as in Example 1.

[0096] (4)组装覆盖层 [0096] (4) assembling the cover layer

[0097] 本实施例制备的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系中,PDMAEA 层、rH-SIA 层、GOD 层在基底上的重叠顺序为:(PDMAEA/rH_SIA/PDMAEA/GOD) 2。 Long-term control of blood glucose concentration of the drug controlled release systems [0097] The present contains insulin aggregates prepared in Example, PDMAEA layer, rH-SIA layer, overlapping order GOD layer on the substrate is: (PDMAEA / rH_SIA / PDMAEA / GOD) 2.

[0098] 按照上述结构,在室温、常压下将步骤(2)配制的PDMAEA溶液100 μ L旋涂于经过表面处理的云母片表面,吸附60min,然后透析处理多余的PDMAEA溶液;将步骤(2)配制的rH-SIA悬浊液100 μ L涂覆于PDMAEA层表面,吸附5min,然后用去离子水清洗表面,并用氮气吹干;将步骤⑵配制的PDMAEA溶液100 μ L旋涂于rH-SIA层表面,吸附20min,然后透析处理多余的PDMAEA溶液;将步骤(2)配制的GOD溶液200 μ L旋涂于PDMAEA层表面,吸附lOmin,然后离心处理多余GOD溶液,得到I个PDMAEA/rH_SIA/PDMAEA/GOD四分子层。 [0098] According to the above structure, at room temperature, the pressure in step (2) was prepared PDMAEA 100 μ L was spin-coated mica surface through surface treatment, adsorption 60min, then excess PDMAEA dialysis treatment solution; in step ( 2) suspension of formulated rH-SIA 100 μ L PDMAEA applied to the surface layer, the adsorption 5min, then the surface rinsed with deionized water, and blown dry with nitrogen; ⑵ formulated PDMAEA step 100 μ L was spin-coated rH -SIA surface layer, the adsorption 20min, then excess PDMAEA dialysis treatment solution; step (2) preparation of GOD was 200 μ L PDMAEA spin-coated layer surface, adsorption lOmin, then the excess GOD solution was centrifuged to obtain the I PDMAEA / rH_SIA / PDMAEA / GOD four molecular layers. 重复上述操作,得到基底上形成了下述覆盖顺序的“(PDMAEA/rH-SIA/PDMAEA/GOD)2”的8层覆盖层,即本实施例所述结构的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系。 Above operation is repeated to give eight layers forming a covering layer covering the following order "(PDMAEA / rH-SIA / PDMAEA / GOD) 2" on the substrate, i.e., the structure of a carrier aggregate long-acting insulin present embodiment control of blood glucose concentration of drug release system.

[0099] 实施例5 [0099] Example 5

[0100] 本实施例的工艺步骤如下: [0100] Process steps of the present embodiment is as follows:

[0101] (I)制备猪胰岛素聚集体 [0101] (I) Preparation of porcine insulin aggregates

[0102] 猪胰岛素聚集体(在下述描述中简称P-SIA)的制备方法与实施例1相同。 Preparation Method [0102] porcine insulin aggregates (referred to in the following description P-SIA) in the same manner as in Example 1.

[0103] (2)配制原料溶液 [0103] (2) Preparation of raw material solution

[0104] 将分子量为IOOOkda的线型聚合物:聚丙烯酸二甲氨基乙酯(在下述描述中简称PDMAEA)用pH值为5.2的盐酸水溶液配制成浓度1.5mg/mL的溶液,然后加入NaCl,NaCl的加入量以其在混合液中的浓度达到0.3mol/L为限; [0104] The molecular weight of the linear polymer IOOOkda: aqueous hydrochloric acid dimethylaminoethyl ester of polyacrylic acid (referred to in the following description PDMAEA) formulated with a pH of 5.2 to a concentration of 1.5mg / mL was then added NaCl, NaCl is added in an amount such that its concentration in the mixture reaches 0.3mol / L is limited;

[0105] 将将步骤(I)制备的P-SIA用去离子水配制成浓度为2mg/mL的P-SIA悬浊液; [0105] The step (I) P-SIA prepared with deionized water to a concentration of 2mg / mL suspension of the P-SIA;

[0106] 将葡萄糖氧化酶(在下述描述中简称GOD)粉末用pH值为6的盐酸水溶液配制成浓度为2mg/mL的GOD溶液; [0106] Glucose oxidase (GOD referred to in the following description) aqueous hydrochloric acid with a pH value of 6 powders were formulated as GOD solution 2mg / mL of;

[0107] (3)处理基底 [0107] (3) treating the substrate

[0108] 基底为聚氨酯材质的导管表面,处理方法同实施例1。 [0108] substrate is a surface of the polyurethane catheter material, the same treatment as in Example 1.

[0109] (4)组装覆盖层 [0109] (4) assembling the cover layer

[0110] 本实施例制备的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系中,PDMAEA 层、P-SIA 层、GOD 层在基底上的重叠顺序为:PDMAEA/P_SIA/PDMAEA/GOD。 [0110] Drug release system according to the present embodiment of the carrier prepared in Example aggregate long-acting insulin to control blood glucose concentrations, PDMAEA layer, P-SIA layer, overlapping order GOD layer on the substrate is: PDMAEA / P_SIA / PDMAEA / GOD. [0111] 按照上述结构,在室温、常压下将步骤(2)配制的PDMAEA溶液150 μ L旋涂于经过表面处理的导管表面,吸附20min,然后透析处理多余的PDMAEA溶液;将步骤(2)配制的P-SIA悬浊液100 μ L旋涂于PDMAEA层表面,吸附15min,然后用去离子水清洗表面,并用氮气吹干;将步骤⑵配制的PDMAEA溶液200 μ L旋涂于P-SIA层表面,吸附lOmin,然后透析处理多余的PDMAEA溶液;将步骤(2)配制的GOD溶液200 μ L旋涂于PDMAEA层表面,吸附lOmin,然后离心处理多余GOD溶液。 [0111] According to the above structure, at room temperature, the pressure in step (2) was prepared PDMAEA 150 μ L was spin-coated catheter surface after surface treatment, adsorption 20min, then excess PDMAEA dialysis treatment solution; Step (2 ) suspension formulated P-SIA 100 μ L PDMAEA spin-coated on the surface layer, the adsorption 15min, and then surface washed with deionized water, and blown dry with nitrogen; ⑵ formulated PDMAEA step 200 μ L was spin-coated P- SIA surface layer, the adsorption lOmin, then the excess PDMAEA dialysis treatment solution; step (2) preparation of GOD was 200 μ L PDMAEA spin-coated layer surface, adsorption lOmin, then centrifuged GOD excess solution. 得到基底上形成了下述覆盖顺序的“PDMAEA/P-SIA/PDMAEA/GOD”的4层覆盖层,即本实施例所述结构的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系。 Formed cover the following sequence "PDMAEA / P-SIA / PDMAEA / GOD" 4-layer covers the substrate layer obtained, i.e., long-term control of blood glucose concentration of the drug release carrier according to the present embodiment of the structure of the insulin aggregates in Example system.

[0112] 实施例6 [0112] Example 6

[0113] 本实施例的工艺步骤如下: [0113] Process steps of the present embodiment is as follows:

[0114] (I)制备猪胰岛素聚集体 [0114] (I) Preparation of porcine insulin aggregates

[0115] 猪胰岛素聚集体(在下述描述中简称P-SIA)的制备方法与实施例1相同。 Preparation Method [0115] porcine insulin aggregates (referred to in the following description P-SIA) in the same manner as in Example 1.

[0116] (2)配制原料溶液 [0116] (2) Preparation of raw material solution

[0117] 将分子量为25kda的壳聚糖用用pH值为5.2乙酸水溶液配制成浓度为2mg/mL的壳聚糖溶液; [0117] The molecular weight of 25kda chitosan with acetic acid aqueous solution with a pH of 5.2 were formulated as chitosan solution 2mg / mL of;

[0118] 将步骤(I )制备的P-SIA用pH值为5的盐酸溶液配制成浓度lmg/mL的P-SIA悬浊液; [0118] The step (I) P-SIA prepared hydrochloric acid solution with a pH value of 5 is formulated at a concentration lmg / mL suspension of the P-SIA;

[0119] 将葡萄糖氧化酶(在下述描述中简称GOD)粉末溶解于pH为5的盐酸溶液中,配成浓度为lmg/mL的溶液。 [0119] Glucose oxidase (referred to in the following description GOD) powder was dissolved in hydrochloric acid solution to pH 5, formulated at a concentration of lmg / mL solution.

[0120] (3)制备胰岛素聚集体基底 [0120] (3) insulin aggregates prepared substrate

[0121] 将步骤(I)制备的P-SIA用pH值为5的盐酸溶液配制成浓度为25mg/mL的P-SIA悬浊液,然后用浓度2N的NaOH溶液调节B-SIA悬浊液的pH值至8,然后加入NaCl,NaCl的加入量以混合液中NaCl的浓度达到4N为限,在室温、常压下搅拌混合8h,继后以3000转/分的转速离心5min,获带负电荷的颗粒状P-SIA基底。 [0121] The step (I) P-SIA prepared solution of hydrochloric acid with a pH value of 5 is formulated at a concentration of 25mg / mL suspension of the P-SIA, then adjust the suspension B-SIA with 2N NaOH solution of concentration to pH 8, followed by addition of NaCl, NaCl is added in an amount to achieve a concentration of NaCl mixture 4N limited to, stirring and mixing at room temperature for 8h, atmospheric pressure, followed by the 3000 r / min centrifuged 5min, obtained with negatively charged P-SIA particulate substrate.

[0122] (4)组装包覆层 [0122] (4) assembling the cladding layer

[0123] 本实施例制备的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系中,壳聚糖层、P-SIA层、GOD层在颗粒状P-SIA基底上的包覆顺序为:(壳聚糖/GOD/壳聚糖/P-SIA)4。 Long-term control of blood glucose concentration of the drug release system containing insulin aggregates [0123] Preparation Example of the present embodiment, the chitosan layer, P-SIA layer, on the order of GOD layer coated particulate substrate P-SIA chitosan is :( / GOD / chitosan / P-SIA) 4.

[0124] 按照上述结构,在室温、常压下将步骤(3)制备的颗粒状P-SIA基底IOmg与步骤(2)配制的壳聚糖溶液5mL通过搅拌的方式混合,搅拌时间60min,搅拌时间届满后,以3000转/分的转速离心5min,然后将所获沉淀用浓度2mM的HCl与浓度0.7M的NaCl混合液洗涤、离心2次,获包覆有壳聚糖层的微粒;在室温、常压下将包覆有壳聚糖层的微粒与步骤 [0124] According to the above structure, at room temperature, the pressure 5mL chitosan solution in step (3) P-SIA particulate substrate prepared in Step IOmg (2) prepared by way of mixing with stirring, mixing time 60min, stirring after the time expires, at 3000 rev / min centrifuged 5min, then the resulting precipitate was washed with a mixture of NaCl with a concentration of 2mM HCl to a concentration of 0.7M, centrifuged twice coated fine particles obtained chitosan layer; in at room temperature, at atmospheric pressure and a step of chitosan particles coated with layer

(2)配制的GOD溶液3mL通过搅拌的方式混合,搅拌时间IOmin,搅拌时间届满后,以3000转/分的转速离心5min,然后将所获沉淀用浓度2mM的HCl与浓度0.7M的NaCl混合液洗涤、离心2次,获依次包覆有壳聚糖层、GOD层的微粒;在室温、常压下将包覆有壳聚糖层、GOD层的微粒与步骤(2)配制的壳聚糖溶液5mL通过搅拌的方式混合,搅拌时间20min,搅拌时间届满后,以3000转/分的转速离心5min,然后将所获沉淀用浓度2mM的HCl与浓度0.7M的NaCl混合液洗涤、离心2次,获依次包覆有壳聚糖层、GOD层、壳聚糖层的微粒;在室温、常压下将包覆有壳聚糖层、GOD层、壳聚糖层的微粒与步骤(2)配制的P-SIA悬浊液2mL通过搅拌的方式混合,搅拌时间15min,搅拌时间届满后,以3000转/分的转速离心5min,然后将所获沉淀用浓度2mM的HCl与浓度0.7M的NaCl混合液洗涤、离心2次,得到I个壳聚糖/GOD/壳聚糖/P-SIA四分子 (2) Preparation of GOD was mixed by stirring 3mL manner, stirring time IOmin, stirring time after the expiration of, at 3000 rpm / min centrifuged 5min, then the resulting precipitate was mixed with a concentration of 2mM HCl to NaCl concentration of 0.7M washing solution, centrifugation twice, was sequentially coated with a layer of chitosan, GOD fine particle layer; the layer is coated with chitosan at room temperature, atmospheric pressure, fine particles and the step of GOD layer (2) prepared chitosan 5mL stirred sugar solution by way of the mixing, mixing time 20min, stirring time after the expiration of, at 3000 rpm / min centrifuged 5min, then the resulting precipitate was washed with a mixture of NaCl with a concentration of 2mM HCl to a concentration of 0.7M, centrifuged for 2 times, was sequentially coated with a chitosan layer, a layer of GOD, chitosan microparticles layer; the layer is coated with chitosan at room temperature, atmospheric pressure, of GOD layer, a layer of chitosan microparticles of step (2 ) P-SIA formulated 2mL suspension by way of mixing with stirring, the stirring time and centrifuged 15min, stirring time after the expiration of, at 3000 rev / min 5min, then the precipitate was obtained by concentration of 2mM HCl to a concentration of 0.7M mixture was washed with NaCl, centrifuged twice, to obtain a chitosan I / GOD / chitosan / P-SIA tetrad . 重复上述操作,在基底上形成了下述包覆顺序的“(壳聚糖/GOD/壳聚糖/P-SIA)/'的16层包覆层,即本实施例所述结构的载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,将所制得的微球状药物控释体系在_80°C冻干干燥10h。 Repeating the above operation, the coating formed the following sequence "(chitosan / of GOD / chitosan / P-SIA) on a substrate / apos cladding layer 16, i.e., the carrier of the structure according to the present embodiment has long-term control of blood glucose concentration of the drug release system of insulin aggregates the resulting microspheres in controlled drug delivery system was dried lyophilized _80 ° C 10h.

[0125] 实施例7 [0125] Example 7

[0126] 动物实验采用健康8周SD大鼠,体重210〜240g。 [0126] Animal experiments using healthy 8 weeks SD rats weighing 210~240g. SD大鼠共20只,随机分为两组,每组10只,其中一组为试验组,一组为健康对照组。 20 SD rats were randomly divided into two groups of 10 each, one set of the test group, a group of healthy controls.

[0127] 将试验组大鼠禁食12h后腹腔注射用柠檬酸缓冲液以4mg/mL的浓度冰浴溶解的链脲佐菌素,按空腹体重50mg/kg注射构建糖尿病大鼠模型,注射一周后空腹血糖值大于16.7mmol/L( = 300mg/dl)的大鼠为患糖尿病的大鼠。 [0127] A test group of rats fasted for 12h after intraperitoneal injection of citrate buffer at 4mg / mL, a concentration of dissolved ice bath streptozotocin, by fasting weight 50mg / kg injected construct diabetic rats, injected one week fasting blood glucose level greater than 16.7mmol / L (= 300mg / dl) in rats diabetic rat infestation. 在患糖尿病大鼠皮下埋植实施例1制备的载有猪胰岛素聚集体的药物控释体系,所有大鼠全天自由取水和饲料。 In diabetic rat subcutaneous implants containing porcine insulin aggregates prepared in Example 1 drug release system embodiment, all rats day free water and feed.

[0128] 监测大鼠血糖的方法是将大鼠尾静脉取出的血用一触式血糖监测系统(OneTouch® UltraEasy™, LifeScan, Inc.Milpitas, CA.)测出。 [0128] The method of monitoring of blood glucose is removed the tail vein of rats using a blood glucose monitoring system touch (OneTouch® UltraEasy ™, LifeScan, Inc.Milpitas, CA.) measured.

[0129] 试验结果见图3,从图3可以看出,使用本发明所述药物控释体系,单次用药可以控制患糖尿病大鼠体内的正常血糖浓度达295天。 [0129] The results shown in Figure 3, it can be seen from Figure 3, the present invention is the use of a controlled release medicament system, a single dose may control normal blood glucose levels in diabetic rats suffering 295 days.

Claims (9)

  1. 1.一种载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,其特征在于由基底及重叠或包覆在基底上的具有PH敏感性的聚合物层和胰岛素聚集体层组成,基底、具有pH敏感性的聚合物层、胰岛素聚集体层相互之间通过静电作用结合成一体; 或由基底及重叠或包覆在基底上的具有PH敏感性的聚合物层、蛋白质层和胰岛素聚集体层组成,基底、具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层相互之间通过静电作用结合成一体; 所述基底为平板、微球或医疗器械,所述平板或微球由有机玻璃、石英、云母中的一种制作。 A controlled drug delivery system depot carrying the control of blood glucose concentrations of insulin aggregates, characterized by the overlapping or covering the substrate and a polymer layer on the substrate and insulin sensitivity PH aggregate layer having a composition, a substrate, having a pH-sensitive polymer layer, between insulin aggregate layer integrally bonded to each other by electrostatic action; or overlapping or by a base and coated on a substrate having a pH sensitive polymer layer, the protein layer and insulin aggregate layers, the substrate, having a PH sensitive polymer layer, the protein layer, between insulin aggregate layer integrally bonded to each other by electrostatic action; the substrate is a tablet, microspheres or medical device, the plate or micro ball made of glass, quartz, mica kind.
  2. 2.根据权利要求1所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,其特征在于具有PH敏感性的聚合物层和胰岛素聚集体层在基底上的包覆顺序可以是具有pH敏感性的聚合物层、胰岛素聚集体层的所有组合方式,或具有PH敏感性的聚合物层、蛋白质层和胰岛素聚集体层在基底上的包覆顺序可以是具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层的所有组合方式。 The drug release system of the long-term control of blood glucose concentration of insulin contained in an aggregate as claimed in claim, characterized in that the coating sequence having a PH sensitive polymer layer and the insulin aggregate layer on the substrate may be pH-sensitive polymer layer, the layer of aggregated all insulin precursor combinations, or with a pH sensitive polymer layer, the protein layer covering the aggregate, and insulin in order on the substrate layer may be a polymeric pH sensitive layer, a protein layer, all combinations of insulin aggregate layer.
  3. 3.一种载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,其特征在于由基底及包覆在基底上的具有PH敏感性的聚合物层和蛋白质层组成,基底、具有pH敏感性的聚合物层、蛋白质层相互之间通过静电作用结合成一体; 或由基底及包覆在基底上的具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层组成,基底、具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层相互之间通过静电作用结合成一体; 所述基底为胰岛素聚集体颗粒。 An aggregate containing insulin control of blood glucose concentration of the drug depot release system, wherein the substrate is coated on a substrate and a polymer layer and a protein layer having a PH sensitive composition, a substrate having a pH sensitive polymer layer between the protein layers are integrally bonded by electrostatic interaction; or a coated substrate and a polymer layer on the substrate having a PH sensitive, protein layer, insulin aggregate layers, the substrate, having PH-sensitive polymer layer, a protein layer, insulin aggregate layer integrally joined to each other by electrostatic interaction; the substrate is insulin aggregate particle.
  4. 4.根据权利要求3所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,其特征在于具有PH敏感性的聚合物层和蛋白质层在基底上的包覆顺序可以是具有pH敏感性的聚合物层、蛋白质层的所有组合方式,或具有PH敏感性的聚合物层、蛋白质层和胰岛素聚集体层在基底上的包覆顺序可以是具有PH敏感性的聚合物层、蛋白质层、胰岛素聚集体层的所有组合方式。 The drug release system of the long-term control of blood glucose concentration of insulin contained 3 aggregates as claimed in claim, characterized in that the coating sequence having a PH sensitive polymer layer and a protein layer on the substrate may have a pH sensitive polymer layer, all combinations of the protein layer, or a polymer having a PH sensitive layers, cladding layers and an insulin protein sequence of the aggregate layer on the substrate may be a polymer layer having a PH sensitive protein layer, all combinations of insulin aggregate layer.
  5. 5.根据权利要求1至4中任一权利要求所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,其特征在于胰岛素聚集体为牛胰岛素聚集体、重组人胰岛素聚集体、猪胰岛素聚集体中的一种。 5. aggregates bovine insulin, human recombinant insulin aggregate according to any one of 1 to 4 claims containing the long-acting control of blood glucose concentration of the drug release system of insulin aggregates, wherein the insulin aggregates, one kind of porcine insulin aggregated body.
  6. 6.根据权利要求1至4中任一权利要求所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,其特征在于具有pH敏感性的聚合物为聚甲基丙烯酸-N,N-二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯、聚甲基丙烯酸酯、壳聚糖中的一种,分子量为I〜lOOOkda。 1 to 4 according to any one of claims depot carrying the blood glucose concentration control drug release system of insulin aggregates, wherein the pH-sensitive polymer is polymethacrylic acid -N, N- dimethylaminoethyl methacrylate, poly-acrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate, polymethyl methacrylate, a chitosan, a molecular weight of I~lOOOkda.
  7. 7.根据权利要求1至4中任一权利要求所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系,其特征在于蛋白质为葡萄糖氧化酶或过氧化氢酶。 1 to 4 according to any one of claims depot carrying the blood glucose concentration control drug release system of insulin aggregates, wherein the protein is a glucose oxidase or catalase.
  8. 8.—种权利要求1或2所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系的制备方法,其特征在于工艺步骤如下: (I)配制原料溶液或悬浊液原料溶液或悬浊液为具有PH敏感性的聚合物溶液和胰岛素聚集体悬浊液,或具有pH敏感性的聚合物溶液、胰岛素聚集体悬浊液和蛋白质溶液;所述具有pH敏感性的聚合物为聚甲基丙烯酸-N,N-二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯、聚甲基丙烯酸酯、壳聚糖中的一种,分子量为I〜IOOOkda ;用pH值为I〜7的盐酸水溶液、乙酸溶液或去离子水将具有pH敏感性的聚合物配制成浓度为0.015〜lOOmg/mL的溶液,若具有pH敏感性的聚合物为聚甲基丙烯酸-N,N-二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯或聚甲基丙烯酸酯,则再加入NaCl,NaCl的加入量以其在混合液中的浓度达到0. The method of preparing a long-term control of blood glucose concentration of the drug release system containing 1 or 2 kinds of insulin aggregates 8.- claims, characterized in that the process steps are as follows: (I) a solution or suspension prepared raw material solution a polymer solution or suspension and a suspension having aggregates pH insulin sensitivity, or with a pH-sensitive polymer solution, suspension and insulin aggregate protein solution; the pH-sensitive polymer as polymethacrylic acid -N, N- dimethylaminoethyl acrylate, polymethacrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate, polymethyl methacrylate, a chitosan, molecular weight I~IOOOkda; I~7 with a pH of aqueous hydrochloric acid, acetic acid solution or deionized water having a pH-sensitive polymer formulation at a concentration of 0.015~lOOmg / mL solution, if the pH-sensitive polymeric was as polymethacrylic acid -N, N- dimethylaminoethyl acrylate, polymethacrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate or polymethyl methacrylate, is then added NaCl, NaCl is amount of its concentration in the mixture reaches zero. 01〜5mol/L为限; 所述胰岛素聚集体为牛胰岛素聚集体、重组人胰岛素聚集体、猪胰岛素聚集体中的一种;用PH值为3〜8的盐酸水溶液、乙酸水溶液、磷酸水溶液或去离子水将胰岛素聚集体配制成浓度为0.015〜100mg/mL的悬池液; 所述蛋白质为葡萄糖氧化酶或过氧化氢酶;用PH值为3〜8的盐酸水溶液、乙酸水溶液、磷酸水溶液或去离子水将蛋白质配制成浓度为0.015〜100mg/mL的溶液; (2)处理基底表面使其带电荷; (3)组装重叠层或包覆层将具有PH敏感性的聚合物溶液、胰岛素聚集体悬浊液和/或蛋白质溶液用旋涂、浸涂或喷涂的方法按照所述药物控释体系的结构重叠或包覆在基底上和在前的重叠或包覆物上,在制备每一层重叠或包覆物时,静置O-lOOmin,并将形成所述重叠或包覆物的多余溶液或悬浊液用去离子水冲洗、离心或透析的方法去掉并干燥,即得到载有胰 01~5mol / L is limited; the insulin is bovine insulin aggregates aggregates aggregates of recombinant human insulin, porcine insulin aggregates of one; with a PH value 3~8 aqueous hydrochloric acid, aqueous acetic acid, aqueous phosphoric acid deionized water or insulin aggregates were formulated as a liquid suspension cell 0.015~100mg / mL; and the protein is a glucose oxidase or catalase; with a PH value 3~8 aqueous hydrochloric acid, aqueous acetic acid, phosphoric acid aqueous solution or deionized water to a protein concentration of formulated 0.015~100mg / mL of solution; (2) treating the substrate surface so as charged; (3) assembling the cladding layer or overlapping layers having a PH sensitive polymer solution, insulin suspension aggregates and / or protein solution by spin-coating, dipping or spraying method or coated on the substrate to overlap on the front and overlap or wrap structure according to the drug release system, in the preparation of when each layer overlap or wrap, allowed to stand O-lOOmin, and forming the wrap overlapping or excess solution or suspension rinsed with deionized water, centrifugation or dialysis method of removing and dried to obtain carrying pancreas 素聚集体的长效控制血糖浓度的药物控释体系; 所述基底为平板、微球或医疗器械,所述平板或微球由有机玻璃、石英、云母中的一种制作。 Long-term control of blood glucose element aggregate concentration of drug controlled release systems; said substrate is a tablet, microspheres or medical device, the plate or microspheres of glass, quartz, mica method of making.
  9. 9.一种权利要求3或4所述载有胰岛素聚集体的长效控制血糖浓度的药物控释体系的制备方法,其特征在于工艺步骤如下: (1)配制原料溶液或悬浊液原料溶液或悬浊液为具有PH敏感性的聚合物溶液和蛋白质溶液,或具有pH敏感性的聚合物溶液、蛋白质溶液和胰岛素聚集体悬浊液; 所述具有PH敏感性的聚合物为聚甲基丙烯酸-N,N-二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯、聚甲基丙烯酸酯、壳聚糖中的一种,分子量为I〜IOOOkda ;用pH值为I〜7的盐酸水溶液、乙酸溶液或去离子水将具有pH敏感性的聚合物配制成浓度为0.015〜lOOmg/mL的溶液,若具有pH敏感性的聚合物为聚甲基丙烯酸-N,N-二甲氨基乙酯、聚甲基丙烯酰胺丙基二甲基胺、聚丙烯酸二甲氨基乙酯或聚甲基丙烯酸酯,则再加入NaCl,NaCl的加入量以其在混合液中的浓度达到0.01〜5mol/L The method of preparing a long-term control of blood glucose concentration of the drug release system or carrier 34 An insulin aggregates claims, characterized in that the process steps are as follows: (1) Preparation of raw material solution a solution or suspension pH-sensitive or suspension with a polymer solution and a protein solution, or a pH-sensitive polymer solution, the protein solution and a suspension of insulin aggregates; pH sensitive polymer having poly methyl acrylic -N, N- dimethylaminoethyl acrylate, polymethacrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate, polymethyl methacrylate, a chitosan, a molecular weight of I~ IOOOkda; I~7 with the pH of the aqueous solution of hydrochloric acid, acetic acid solution or deionized water having a pH-sensitive polymer formulation at a concentration of 0.015~lOOmg / mL solution, if having a pH-sensitive polymer is poly a methacrylic acid -N, N- dimethylaminoethyl acrylate, polymethacrylamide dimethylamine, polyacrylic acid, dimethylaminoethyl methacrylate or polymethyl methacrylate, NaCl is added, its addition amount of NaCl concentration in the mixture reaches 0.01~5mol / L 为限; 所述胰岛素聚集体为牛胰岛素聚集体、重组人胰岛素聚集体、猪胰岛素聚集体中的一种;用PH值为3〜8的盐酸水溶液、乙酸水溶液、磷酸水溶液或去离子水将胰岛素聚集体配制成浓度为0.015〜100mg/mL的悬池液; 所述蛋白质为葡萄糖氧化酶或过氧化氢酶;用PH值为3〜8的盐酸水溶液、乙酸水溶液、磷酸水溶液或去离子水将蛋白质配制成浓度为0.015〜100mg/mL的溶液; (2)制备胰岛素聚集体基底将胰岛素聚集体用pH 2〜5的乙酸或盐酸溶液配制成浓度为I〜100mg/mL悬池液,然后用NaOH溶液调节聚集体悬浊液的pH值至6〜10,然后加入NaCl,NaCl的加入量以混合液中NaCl的浓度达到0.5〜5mol/L,在室温、常压下搅拌混合I〜20h,继后离心分离,获带负电荷的颗粒状胰岛素聚集体基底; (3)组装包覆层将具有PH敏感性的聚合物溶液和蛋白质溶液采用与基底混合和与在前的包覆物 Limited; the insulin is bovine insulin aggregates aggregates aggregates of recombinant human insulin, porcine insulin aggregates of one; with a PH value 3~8 aqueous hydrochloric acid, aqueous acetic acid, aqueous phosphoric acid solution or deionized water insulin aggregates were formulated as a liquid suspension cell 0.015~100mg / mL; and the protein is a glucose oxidase or catalase; with a PH value 3~8 aqueous hydrochloric acid, aqueous acetic acid, aqueous phosphoric acid solution or deionized water the protein was formulated at a concentration of 0.015~100mg / mL of solution; (2) preparing an insulin basal insulin aggregates aggregates pH 2~5 with acetic acid or hydrochloric acid solutions were prepared at a concentration of I~100mg / mL cell suspension was then aggregates adjusted with NaOH solution to pH 6~10 suspension, followed by addition of NaCl, NaCl is added in an amount to achieve a concentration of NaCl mixture 0.5~5mol / L, mixing I~20h stirred at room temperature, atmospheric pressure , following centrifugation, is eligible for a negatively charged insulin aggregate particulate substrate; (3) assembling the cladding layer having a PH sensitive protein solution and polymer solution mixed with the substrate and with the use of previous wrap 混合的方法按照所述药物控释体系的结构包覆在基底上和在前的包覆物上,或将具有PH敏感性的聚合物溶液、蛋白质溶液和胰岛素聚集体悬浊液采用与基底混合和与在前的包覆物混合的方法按照所述药物控释体系的结构包覆在基底上和在前的包覆物上,在制备每一层包覆物时,搅拌至少lOmin,并将形成所述包覆物的多余溶液或悬浊液用去离子水冲洗、离心或透析的方法去掉,即得到载有胰岛素聚集体的长效控制血糖浓度的药物控释体系。 The method of mixing the coating on the substrate and covering the front of the controlled drug release composition in accordance with the structure of the system, or a polymer solution having a PH sensitive, protein aggregates, and insulin solution is mixed with the substrate using a suspension and was mixed with the former method of coating structure according to the drug release system is coated on the upper substrate and the front coating was coated in the preparation of each layer was stirred for lOmin at least, and excess solution or suspension forming the coating was rinsed with deionized water, centrifugation or dialysis method for removing, i.e., to obtain aggregates containing the long-acting insulin to control blood glucose concentration of drug release system.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4364385A (en) * 1981-03-13 1982-12-21 Lossef Steven V Insulin delivery device
WO1984004932A1 (en) * 1983-06-10 1984-12-20 Washington Res Found Biochemical detection and/or treatment process
US4764364A (en) * 1986-02-25 1988-08-16 S R I International Method of preparing bioerodible polymers having pH sensitivity in the acid range and resulting product
US4855132A (en) * 1986-02-25 1989-08-08 S R I International Method of preparing bioerodible polymers having pH sensitivity in the acid range and resulting product
CN101951957A (en) * 2008-01-04 2011-01-19 百达尔公司 Insulin formulations for insulin release as a function of tissue glucose levels

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4364385A (en) * 1981-03-13 1982-12-21 Lossef Steven V Insulin delivery device
WO1984004932A1 (en) * 1983-06-10 1984-12-20 Washington Res Found Biochemical detection and/or treatment process
US4764364A (en) * 1986-02-25 1988-08-16 S R I International Method of preparing bioerodible polymers having pH sensitivity in the acid range and resulting product
US4855132A (en) * 1986-02-25 1989-08-08 S R I International Method of preparing bioerodible polymers having pH sensitivity in the acid range and resulting product
CN101951957A (en) * 2008-01-04 2011-01-19 百达尔公司 Insulin formulations for insulin release as a function of tissue glucose levels

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