CN103948914A - Method for preparing drug-loadable low-molecular-weight water-soluble chitosan nano-particle - Google Patents
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- 239000002105 nanoparticle Substances 0.000 title claims abstract description 36
- 238000000034 method Methods 0.000 title claims abstract description 7
- 239000007864 aqueous solution Substances 0.000 claims abstract description 27
- 239000000243 solution Substances 0.000 claims abstract description 26
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims abstract description 22
- 229940098773 bovine serum albumin Drugs 0.000 claims abstract description 22
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- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 4
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- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
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Abstract
一种可载药的低分子量水溶性壳聚糖纳米粒子的制备方法，该方法包括如下步骤：a、将水溶性壳聚糖溶于去离子水中配置浓度2-10mg/ml的水溶性壳聚糖溶液；b、依次加入浓度0.1-0.3mg/ml的牛血清蛋白水溶液和浓度0.1-0.3mg/ml的三聚磷酸钠水溶液，室温搅拌条件下，自发生成载药纳米粒子；其中，其中，水溶性壳聚糖溶液、牛血清蛋白水溶液和三聚磷酸钠水溶液的体积比为1：（0.1-0.3）：（0.1-0.3）。本发明在中性条件下获得水溶性壳聚糖纳米颗粒，对牛血清蛋白等药物存在缓释效果，工艺简单可行、所得的纳米粒子平均粒径小。A preparation method of drug-loadable low molecular weight water-soluble chitosan nanoparticles, the method comprising the following steps: a, dissolving water-soluble chitosan in deionized water to prepare water-soluble chitosan with a concentration of 2-10mg/ml sugar solution; b, sequentially add bovine serum albumin aqueous solution with a concentration of 0.1-0.3 mg/ml and sodium tripolyphosphate aqueous solution with a concentration of 0.1-0.3 mg/ml, and spontaneously generate drug-loaded nanoparticles under room temperature stirring conditions; wherein, wherein, The volume ratio of the water-soluble chitosan solution, bovine serum albumin solution and sodium tripolyphosphate solution is 1: (0.1-0.3): (0.1-0.3). The invention obtains water-soluble chitosan nanoparticles under neutral conditions, has slow-release effect on drugs such as bovine serum albumin, has simple and feasible technology, and has small average particle diameter of the obtained nanoparticles.
Description
the
技术领域 technical field
本发明涉及一种可载药的低分子量水溶性壳聚糖纳米粒子的制备方法。 The invention relates to a preparation method of low-molecular-weight water-soluble chitosan nanoparticles that can be loaded with drugs.
背景技术 Background technique
壳聚糖是自然界中唯一的天然的碱性多糖，其基本单位为葡萄糖胺，不溶于水、碱性溶液或普通有机溶剂，但可溶于盐酸、硝酸、甲酸、乙酸等稀酸，壳聚糖以其优良特性，例如生物相容性、降解性、无毒性、吸附性、来源丰富等，已在医药、食品、化工、农业、环保等领域得到广泛应用，并于1991年被日本、美国、欧洲等地的医学界和营养食品研究机构认定为在碳水化合物、蛋白质、脂肪、维生素、矿物质之后的第六大生命要素。 Chitosan is the only natural alkaline polysaccharide in nature, its basic unit is glucosamine, insoluble in water, alkaline solution or common organic solvent, but soluble in hydrochloric acid, nitric acid, formic acid, acetic acid and other dilute acids, chitosan With its excellent properties, such as biocompatibility, degradability, non-toxicity, adsorption, and rich sources, sugar has been widely used in the fields of medicine, food, chemical industry, agriculture, and environmental protection. It was approved by Japan, the United States in 1991 Medical circles and nutritional food research institutions in Europe, Europe and other places have identified it as the sixth major life element after carbohydrates, proteins, fats, vitamins and minerals.
壳聚糖具有生物相容性、无毒性和生物粘附性，可使药物在局部保持高浓度，且可打开上皮粘膜的紧密连接，增强药物在粘膜的渗透作用。壳聚糖纳米粒子比微米粒子具有更多优越性，可使大分子顺利通过上皮组织，促进药物的渗透吸收，适当的表面修饰使它对特定器官或病灶具有靶向作用。尤其适用于口服或粘膜等更为方便的给药途径，可延长药物在体内的循环时间，有效地提高药物的生物利用度，减少副作用。 Chitosan has biocompatibility, non-toxicity and bioadhesion, which can keep the drug at a high concentration locally, and can open the tight junction of the epithelial mucous membrane, and enhance the penetration of the drug in the mucous membrane. Chitosan nanoparticles have more advantages than micro-particles, which can make macromolecules pass through epithelial tissue smoothly, promote the penetration and absorption of drugs, and proper surface modification makes it have a targeting effect on specific organs or lesions. It is especially suitable for more convenient routes of administration such as oral or mucous membranes, which can prolong the circulation time of drugs in the body, effectively improve the bioavailability of drugs, and reduce side effects.
由于常用的壳聚糖分子量大、粘度高，无法在生理pH条件下溶解，限制了壳聚糖作为疫苗载体的应用。而低分子量的壳聚糖及其纳米粒的细胞毒性较低，纳米粒子可显著增加A549细胞的摄取作用。 Due to the large molecular weight and high viscosity of commonly used chitosan, it cannot be dissolved under physiological pH conditions, which limits the application of chitosan as a vaccine carrier. However, low molecular weight chitosan and its nanoparticles have low cytotoxicity, and the nanoparticles can significantly increase the uptake by A549 cells.
目前在制备纳米粒子过程中必须将乙酸作为增溶剂，而乙酸环境对生物活性药物产生了严重的影响。这些缺点主要与壳聚糖的N-乙酰化度、分子量等物化性质有关。 At present, acetic acid must be used as a solubilizer in the process of preparing nanoparticles, and the acetic acid environment has a serious impact on bioactive drugs. These disadvantages are mainly related to physicochemical properties such as N-acetylation degree and molecular weight of chitosan.
发明内容 Contents of the invention
本发明的目的是提供一种工艺简单可行、所得的纳米粒子平均粒径小的可载药的低分子量水溶性壳聚糖纳米粒子的制备方法， The object of the present invention is to provide a kind of process simple and feasible, the preparation method of the low molecular weight water-soluble chitosan nano-particle that can be loaded with medicine that the nano-particle average particle diameter of gained is little,
本发明提供的一种可载药的低分子量水溶性壳聚糖纳米粒子的制备方法，该方法包括如下步骤： A kind of preparation method of the low molecular weight water-soluble chitosan nano-particle that can be loaded with medicine provided by the invention, the method comprises the steps:
a、将水溶性壳聚糖溶于去离子水中配置浓度2-10mg/ml的水溶性壳聚糖溶液； a, dissolving water-soluble chitosan in deionized water to configure a water-soluble chitosan solution with a concentration of 2-10mg/ml;
b、依次加入浓度0.1-0.3mg/ml的牛血清蛋白水溶液和浓度0.1-0.3mg/ml的三聚磷酸钠水溶液，室温搅拌条件下，自发生成载药纳米粒子； b. Add bovine serum albumin aqueous solution with a concentration of 0.1-0.3 mg/ml and sodium tripolyphosphate aqueous solution with a concentration of 0.1-0.3 mg/ml in sequence, and spontaneously generate drug-loaded nanoparticles under stirring conditions at room temperature;
其中，水溶性壳聚糖溶液、牛血清蛋白水溶液和三聚磷酸钠水溶液的体积比为1：（0.1-0.3）：（0.1-0.3）。 Wherein, the volume ratio of the water-soluble chitosan solution, bovine serum albumin solution and sodium tripolyphosphate solution is 1:(0.1-0.3):(0.1-0.3).
在上述中，牛血清蛋白水溶液的浓度优选为0.2mg/ml，三聚磷酸钠水溶液的浓度优选为0.2mg/ml。水溶性壳聚糖溶液、牛血清蛋白水溶液和三聚磷酸钠水溶液的体积比优选为1：0.2：0.2。 Among the above, the concentration of the bovine serum albumin aqueous solution is preferably 0.2 mg/ml, and the concentration of the sodium tripolyphosphate aqueous solution is preferably 0.2 mg/ml. The volume ratio of the water-soluble chitosan solution, bovine serum albumin solution and sodium tripolyphosphate solution is preferably 1:0.2:0.2.
本发明具有的有益效果：在中性条件下获得水溶性壳聚糖纳米颗粒，对牛血清蛋白等药物存在缓释效果，因此，该壳聚糖颗粒有望成为一些药物的缓释载体，在医药研究中用于制备缓释药物制剂。 The present invention has beneficial effects: water-soluble chitosan nanoparticles can be obtained under neutral conditions, and there is a slow-release effect on drugs such as bovine serum albumin. Therefore, the chitosan particles are expected to become slow-release carriers for some drugs. Used in the preparation of sustained-release pharmaceutical preparations in research.
具体实施方式 Detailed ways
下述实施例中，水溶性壳聚糖的分子量为6.3 kDa、脱乙酰度为90.7%；配制牛血清蛋白水溶液的牛血清蛋白的分子量为68kDa。 In the following examples, the molecular weight of water-soluble chitosan is 6.3 kDa, and the degree of deacetylation is 90.7%; the molecular weight of the bovine serum albumin for preparing the bovine serum albumin aqueous solution is 68kDa.
实施例1 Example 1
将水溶性壳聚糖溶于去离子水中配成5 ml的2mg/ml的水溶性壳聚糖溶液。在室温和磁力搅拌条件下，依次加入浓度0.2mg/ml的牛血清蛋白水溶液1ml和浓度0.2mg/ml的三聚磷酸钠水溶液1ml，室温搅拌条件下，自发生成载药纳米粒子。所得的载药纳米粒子的平均粒径为135.5±10.2 nm。 Water-soluble chitosan was dissolved in deionized water to be made into 5 ml of 2mg/ml water-soluble chitosan solution. Under the condition of room temperature and magnetic stirring, 1 ml of bovine serum albumin aqueous solution with a concentration of 0.2 mg/ml and 1 ml of sodium tripolyphosphate aqueous solution with a concentration of 0.2 mg/ml were sequentially added, and the drug-loaded nanoparticles were spontaneously generated under the condition of stirring at room temperature. The average particle size of the obtained drug-loaded nanoparticles was 135.5±10.2 nm.
实施例2 Example 2
将水溶性壳聚糖溶于去离子水中配成5 ml的2.8mg/ml的水溶性壳聚糖溶液。在室温和磁力搅拌条件下，依次加入浓度0.2mg/ml的牛血清蛋白水溶液1ml和浓度0.2mg/ml的三聚磷酸钠水溶液1ml，室温搅拌条件下，自发生成载药纳米粒子。所得的载药纳米粒子的平均粒径为99.7±9.4 nm。 Water-soluble chitosan was dissolved in deionized water to be made into 5 ml of 2.8mg/ml water-soluble chitosan solution. Under the condition of room temperature and magnetic stirring, 1 ml of bovine serum albumin aqueous solution with a concentration of 0.2 mg/ml and 1 ml of sodium tripolyphosphate aqueous solution with a concentration of 0.2 mg/ml were sequentially added, and the drug-loaded nanoparticles were spontaneously generated under the condition of stirring at room temperature. The average particle size of the obtained drug-loaded nanoparticles was 99.7±9.4 nm.
实施例3 Example 3
将水溶性壳聚糖溶于去离子水中配成5 ml的3.6mg/ml的水溶性壳聚糖溶液。在室温和磁力搅拌条件下，依次加入浓度0.2mg/ml的牛血清蛋白水溶液1ml和浓度0.2mg/ml的三聚磷酸钠水溶液1ml，室温搅拌条件下，自发生成载药纳米粒子。所得的载药纳米粒子的平均粒径为110.2±11.3 nm。 Water-soluble chitosan was dissolved in deionized water to be made into 5 ml of 3.6mg/ml water-soluble chitosan solution. Under the condition of room temperature and magnetic stirring, 1 ml of bovine serum albumin aqueous solution with a concentration of 0.2 mg/ml and 1 ml of sodium tripolyphosphate aqueous solution with a concentration of 0.2 mg/ml were sequentially added, and the drug-loaded nanoparticles were spontaneously generated under the condition of stirring at room temperature. The average particle size of the obtained drug-loaded nanoparticles was 110.2±11.3 nm.
实施例4 Example 4
将水溶性壳聚糖溶于去离子水中配成5 ml的4mg/ml的水溶性壳聚糖溶液。在室温和磁力搅拌条件下，依次加入浓度0.2mg/ml的牛血清蛋白水溶液1ml和浓度0.2mg/ml的三聚磷酸钠水溶液1ml，室温搅拌条件下，自发生成载药纳米粒子。所得的载药纳米粒子的平均粒径为55.1±8.7 nm。 Water-soluble chitosan was dissolved in deionized water to be made into 5 ml of 4mg/ml water-soluble chitosan solution. Under the condition of room temperature and magnetic stirring, 1ml of bovine serum albumin aqueous solution with a concentration of 0.2mg/ml and 1ml of sodium tripolyphosphate aqueous solution with a concentration of 0.2mg/ml were sequentially added, and the drug-loaded nanoparticles were spontaneously generated under the condition of stirring at room temperature. The average particle size of the obtained drug-loaded nanoparticles was 55.1±8.7 nm.
实施例5 Example 5
将水溶性壳聚糖溶于去离子水中配成5 ml的6mg/ml的水溶性壳聚糖溶液。在室温和磁力搅拌条件下，依次加入浓度0.2mg/ml的牛血清蛋白水溶液1ml和浓度0.2mg/ml的三聚磷酸钠水溶液1ml，室温搅拌条件下，自发生成载药纳米粒子。所得的载药纳米粒子的平均粒径为127.3±10.7nm。 Water-soluble chitosan was dissolved in deionized water to be made into 5 ml of 6mg/ml water-soluble chitosan solution. Under the condition of room temperature and magnetic stirring, 1 ml of bovine serum albumin aqueous solution with a concentration of 0.2 mg/ml and 1 ml of sodium tripolyphosphate aqueous solution with a concentration of 0.2 mg/ml were sequentially added, and the drug-loaded nanoparticles were spontaneously generated under the condition of stirring at room temperature. The obtained drug-loaded nanoparticles have an average particle diameter of 127.3±10.7nm.
实施例6 Example 6
将水溶性壳聚糖溶于去离子水中配成5 ml的7mg/ml的水溶性壳聚糖溶液。在室温和磁力搅拌条件下，依次加入浓度0.2mg/ml的牛血清蛋白水溶液1ml和浓度0.2mg/ml的三聚磷酸钠水溶液1ml，室温搅拌条件下，自发生成载药纳米粒子。所得的载药纳米粒子的平均粒径为124.7±9.9nm。 Water-soluble chitosan was dissolved in deionized water to be made into 5 ml of 7mg/ml water-soluble chitosan solution. Under the condition of room temperature and magnetic stirring, 1 ml of bovine serum albumin aqueous solution with a concentration of 0.2 mg/ml and 1 ml of sodium tripolyphosphate aqueous solution with a concentration of 0.2 mg/ml were sequentially added, and the drug-loaded nanoparticles were spontaneously generated under the condition of stirring at room temperature. The average particle diameter of the obtained drug-loaded nanoparticles is 124.7±9.9nm.
实施例7 Example 7
将水溶性壳聚糖溶于去离子水中配成5 ml的8mg/ml的水溶性壳聚糖溶液。在室温和磁力搅拌条件下，依次加入浓度0.2mg/ml的牛血清蛋白水溶液1ml和浓度0.2mg/ml的三聚磷酸钠水溶液1ml，室温搅拌条件下，自发生成载药纳米粒子。所得的载药纳米粒子的平均粒径为111.7±8.7nm。 Water-soluble chitosan was dissolved in deionized water to be made into 5 ml of 8mg/ml water-soluble chitosan solution. Under the condition of room temperature and magnetic stirring, 1 ml of bovine serum albumin aqueous solution with a concentration of 0.2 mg/ml and 1 ml of sodium tripolyphosphate aqueous solution with a concentration of 0.2 mg/ml were sequentially added, and the drug-loaded nanoparticles were spontaneously generated under the condition of stirring at room temperature. The average particle diameter of the obtained drug-loaded nanoparticles is 111.7±8.7nm.
实施例8 Example 8
将水溶性壳聚糖溶于去离子水中配成5 ml的10mg/ml的水溶性壳聚糖溶液。在室温和磁力搅拌条件下，依次加入浓度0.2mg/ml的牛血清蛋白水溶液1ml和浓度0.2mg/ml的三聚磷酸钠水溶液1ml，室温搅拌条件下，自发生成载药纳米粒子。所得的载药纳米粒子的平均粒径为135.5±10.2nm。 Water-soluble chitosan was dissolved in deionized water to be made into 5 ml of 10mg/ml water-soluble chitosan solution. Under the condition of room temperature and magnetic stirring, 1 ml of bovine serum albumin aqueous solution with a concentration of 0.2 mg/ml and 1 ml of sodium tripolyphosphate aqueous solution with a concentration of 0.2 mg/ml were sequentially added, and the drug-loaded nanoparticles were spontaneously generated under the condition of stirring at room temperature. The average particle diameter of the obtained drug-loaded nanoparticles is 135.5±10.2nm.
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| CN104906073A (en) * | 2015-06-10 | 2015-09-16 | 青岛大学附属医院 | Preparation method of chitosan quaternary ammonium salt hyaluronic acid nanogel coated with basic fibroblast growth factors |
| CN104958251A (en) * | 2015-06-10 | 2015-10-07 | 杨甫进 | Preparation method of hyaluronic acid nanogel |
| CN111420067A (en) * | 2020-03-09 | 2020-07-17 | 西南交通大学 | A kind of composite microsphere nanocarrier and its preparation method and application |
| CN112245392A (en) * | 2020-10-25 | 2021-01-22 | 台州职业技术学院 | Preparation and application of low-molecular-weight chitosan modified fatty acid drug-loaded vesicle |
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| CN1686560A (en) * | 2005-04-08 | 2005-10-26 | 武汉大学 | Chitin tetra ammonium salt nano-particle, its preparation method and use |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104906073A (en) * | 2015-06-10 | 2015-09-16 | 青岛大学附属医院 | Preparation method of chitosan quaternary ammonium salt hyaluronic acid nanogel coated with basic fibroblast growth factors |
| CN104958251A (en) * | 2015-06-10 | 2015-10-07 | 杨甫进 | Preparation method of hyaluronic acid nanogel |
| CN104958251B (en) * | 2015-06-10 | 2018-05-29 | 青岛市中心医院 | A kind of preparation method of hyaluronic acid nanometer gel |
| CN104906073B (en) * | 2015-06-10 | 2018-08-10 | 青岛大学附属医院 | A kind of preparation method for the chitosan quaternary ammonium salt hyaluronic acid nanometer gel containing basic fibroblast growth factor |
| CN111420067A (en) * | 2020-03-09 | 2020-07-17 | 西南交通大学 | A kind of composite microsphere nanocarrier and its preparation method and application |
| CN112245392A (en) * | 2020-10-25 | 2021-01-22 | 台州职业技术学院 | Preparation and application of low-molecular-weight chitosan modified fatty acid drug-loaded vesicle |
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