CN116392441B - 一种酸性成纤维细胞生长因子滴眼液制剂 - Google Patents
一种酸性成纤维细胞生长因子滴眼液制剂 Download PDFInfo
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- CN116392441B CN116392441B CN202310431075.4A CN202310431075A CN116392441B CN 116392441 B CN116392441 B CN 116392441B CN 202310431075 A CN202310431075 A CN 202310431075A CN 116392441 B CN116392441 B CN 116392441B
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明公开了一种重组人酸性成纤维细胞生长因子制剂，尤其涉及一种酸性成纤维细胞生长因子滴眼剂，本发明的滴眼剂使用选自组氨酸、吐温80和肝素钠的组合作为蛋白保护剂，无需使用人血白蛋白作为蛋白保护剂，不仅降低了成本，而且有效的提高了产品的稳定性，在(25±2)℃、相对湿度(60±5)％的条件下9个月依然保持较高的生物活性。
Description
技术领域
本发明涉及生物医药技术领域，尤其涉及一种稳定的酸性成纤维细胞生长因子制剂。
背景技术
成纤维细胞生长因子(FGFs)是一种多功能强力细胞因子，对促进成纤维细胞的代谢和胶原蛋白的形成发挥着重要功能。纤维细胞生长因子能促进皮肤组织的生长繁殖，它通过与细胞表面特异受体结合，调控皮肤上皮，内皮和基质细胞的分裂、繁殖和生长分化，促进细胞代谢，增强氧化作用；能促进与皮肤损伤有关细胞的迅速生长繁殖，并调节细胞间基质的合成、分泌及分解；能促进角质层细胞的再生，加速皮肤角质层和基质层的修复，促进人体皮肤细胞的生长；能增强皮肤细胞的蛋白质的合成和细胞代谢，具有延缓皮肤细胞衰老、促进表皮细胞的修复和生长作用。
成纤维细胞生长因子滴眼液对来源于中胚层和外胚层的细胞具有促进修复和再生作用。动物试验结果表明，本品对家兔碱烧伤角膜上皮的再生、角膜基质层和内皮层的修复均有促进作用；市售产品包括酸性成纤维细胞生长因子(aFGF或称FGF-1)冻干粉和碱性成纤维细胞生长因子(bFGF或称FGF-2)滴眼液。aFGF冻干粉需在2-8℃避光保存和运输，有效期24个月左右，使用时将包装中所配置的溶媒倒入装有rhaFGF冻干粉的瓶中；bFGF滴眼液可直接使用，但需在2-8℃保存，有效期约为24个月，一旦放置在室温环境需尽快使用。因此对适用于aFGF的稳定剂进行研究，获得稳定的aFGF药物组合物，提高患者使用的依从性和便利性具有重要的意义；本领域迫切需要提供能保持酸性成纤维细胞生长因子长期稳定性的液态制剂。
发明内容
本发明针对现有技术的不足，提供了一种稳定的酸性成纤维细胞生长因子制剂及其制备方法和应用，对该制剂的各成分进行了研究，特别是选择了不同稳定剂组合并对各稳定剂的配比做出比较，经过反复实验研究，得到了稳定的含有酸性成纤维细胞生长因子的制剂。
本发明提供一种人酸性成纤维细胞生长因子滴眼液，所述的滴眼液包含蛋白保护剂、保湿剂、缓冲盐和渗透压调节剂，所述的蛋白保护剂选自组氨酸、吐温80和肝素钠的组合。
本发明提供一种重组人酸性成纤维细胞生长因子滴眼液，每100ml滴眼液包含0.01～0.2g组氨酸、0.01～0.2g吐温80和0.005～0.1g肝素钠；优选的包含0.1～0.2g组氨酸、0.03-0.1g吐温80和0.01～0.05g肝素钠。
本发明提供一种重组人酸性成纤维细胞生长因子滴眼液，每100ml滴眼液包含1～3mg人酸性成纤维细胞生长因子，优选的包含2mg人酸性成纤维细胞生长因子。
本发明提供一种重组人酸性成纤维细胞生长因子滴眼液，所述的保湿剂选自透明质酸钠、硫酸软骨素、聚乙烯醇、聚乙二醇、甘油、羟丙甲纤维素、右旋糖酐、羧甲基纤维素中的一种或多种，每100ml滴眼液包含0.05～1.5g保湿剂，优选的每100ml滴眼液包含0.1g保湿剂。
本发明提供一种重组人酸性成纤维细胞生长因子滴眼液，所述的缓冲盐选自磷酸盐缓冲体系、组氨酸盐缓冲体系、醋酸盐缓冲体系、柠檬酸盐缓冲体系、Tris缓冲体系、HEPES缓冲体系，优选的所述缓冲盐的pH值为6.0-7.0，浓度为0.01-0.05mol/L，优选的浓度0.02mol/L。
本发明提供一种重组人酸性成纤维细胞生长因子滴眼液，所述的渗透压调节剂选自氯化钠、硼酸、葡萄糖中的一种或多种，优选的每100ml滴眼液包含0.5～0.9g渗透压调节剂。
本发明提供一种重组人酸性成纤维细胞生长因子滴眼液，所述的滴眼液还包含抑菌剂。
本发明提供一种重组人酸性成纤维细胞生长因子滴眼液，所述的抑菌剂选自羟苯乙酯、苯扎氯铵、苯扎溴铵、聚季铵盐、硝酸苯汞、硫柳汞、苯扎氯铵、三氯叔丁醇、羟苯酯类、山梨醇中的一种或多种，优选的每100ml滴眼液包含0.01～0.1g抑菌剂。
本发明提供一种重组人酸性成纤维细胞生长因子滴眼液，所述的滴眼剂进一步包含抗氧化剂，所述的抗氧化剂选自硫代硫酸钠、亚硫酸钠、维生素E、抗坏血酸钠、甲硫氨酸中的一种或多种，优选的每100ml滴眼液包含0.01～0.1g抗氧化剂。
本发明提供一种重组人酸性成纤维细胞生长因子滴眼液，所述的滴眼剂进一步包含增稠剂；优选的所述增稠剂选自选自聚乙烯醇、羧甲基纤维素、甲基纤维素、羧甲基纤维素钠、聚丙烯酸钠、聚乙烯吡咯烷酮中的一种或多种；优选的，所述每100ml滴眼液包含0.5～2g增稠剂。
本发明具有以下有益效果：
1.本发明的滴眼液选用组氨酸、吐温80和肝素钠的组合作为蛋白保护剂，有效的提高了产品的稳定性，在(25±2)℃、相对湿度(60±5)％的条件下9个月依然保持较高的生物活性。
2.本发明所使用的组合蛋白保护剂无需使用价格很高的人血白蛋白，在保证产品品质的前提下，有效的降低了生产成本。
3.通过筛选，本发明发现多种优选的抑菌剂和抗氧化剂的组合，有助于协同增加了产品的稳定性。
具体实施方式
为便于更好地理解本发明，通过以下实例加以说明，这些实例属于本发明的保护范围，但不限制本发明的保护范围。
实施例1：本实施例提供一种重组人酸性成纤维细胞生长因子制剂Ⅰ，其配方和制备方法如下：
制备100ml滴眼液：按上述配比，将保湿剂透明质酸钠，蛋白保护剂组氨酸、吐温80、肝素钠，增稠剂聚乙烯醇，防腐剂三氯叔丁醇，抗氧化剂甲硫氨酸置于80ml磷酸盐缓冲液(0.02mol/L的)中；加入0.9g氯化钠，完全溶解后加入人酸性成纤维细胞生长因子。混合均匀后，加入pH值为6.5的浓度为0.02mol/L的磷酸盐缓冲液定容至所需体积，用0.22μm的微孔滤膜过滤除菌，滤液分装至滴眼液塑料瓶中，即制成本发明滴眼液成品。
实施例2：本实施例的制备方法如实施例1，具体的配方如下：
实施例3：本实施例的制备方法如实施例1，具体的配方如下：
实施例4：本实施例的制备方法如实施例1，具体的配方如下：
对比例1：本对比例的制备方法参照实施例1，具体的配方如下：
对比例2：本对比例的制备方法参照实施例1，具体的配方如下：
对比例3：本对比例的制备方法参照实施例1，具体的配方如下：
对比例4：本对比例的制备方法参照实施例1，具体的配方如下：
对比例5：本对比例的制备方法参照实施例1，具体的配方如下：
效果例：加速稳定性试验
将实施例1-4和对比例1-4制备得到的滴眼液置于(25±2)℃、相对湿度(60±5)％的条件下9个月后，然后进行活性测试，活性测试方法如下：
(1)标准品溶液制备：取重组人酸性成纤维细胞生长因子标准品，在无菌条件下按说明书复溶后，稀释至每1ml含40IU。在96孔细胞培养板中，做4倍系列稀释，做8个稀释度，每个稀释度做2个孔。
(2)供试品溶液制备：稀释方法和处理方法如标准品样品。
(3)将上述样品立即进行下述步骤(4)的检测。
(4)活性测定：取Balb/c 3T3细胞株用完全培养液于37℃、5％CO2条件下培养，控制细胞浓度为1.0×105～5.0×105个细胞/ml，传代后24～36h用于生物学活性测定。弃去培养瓶中的培养液，消化并收集细胞，用完全培养液配制成5.0×104～1.3×105个细胞/ml的细胞悬液，接种于96孔细胞培养板中，每孔100μL。在37℃、5％CO2条件下培养24h。之后换为维持培养液，置37℃、5％CO2条件下培养28～32h。制备的细胞培养板弃去维持液，加入标准溶液和供试品，每孔100μL。置37℃、5％CO2条件下培养64～72h。培养结束，每孔加入MTT溶液20μL，置37℃、5％CO2条件下培养5～6h。向每孔中加入裂解液100μL，混匀后放入酶标仪，以630nm为参比波长，于波长570nm处测定吸光度，记录测定结果。
结果计算：试验数据采用计算机程序或直线回归计算法进行处理，并按下式计算结果：
供试品生物学活性(IU/ml)＝Pr×Ds×Es/(Dr×Er)
相对活性(％)＝供试品生物学活性(加速试验后)/供试品生物学活性(稳定性0点)
式中Pr为标准品生物学活性，单位IU/ml；Ds为供试品预稀释倍数；Dr为标准品预稀释倍数；Es为供试品相当于标准品半效量稀释倍数；Er为标准品半效稀释倍数；加速试验后的相对活性(％)检测数据参见表1：
以上数据表明组氨酸、吐温80、肝素钠的组合作为蛋白保护剂的组合有利于提高人酸性成纤维细胞生长因子加速试验条件下的稳定性；对于抗氧化剂和防腐剂对于稳定性的影响，实施例1中三氯叔丁醇和甲硫氨酸的组合是最优的实施方式，其他防腐剂和抗氧化剂的效果稍差一些。
以上内容不能认定本发明具体实施只局限于这些说明，对于本发明所属技术领域的普通技术人员来说，在不脱离本发明构思前提下，还可以做出若干简单推演或替换，都应当视为属于本发明由所提交的权利要求书确定的专利保护范围。
Claims (8)
1.一种重组人酸性成纤维细胞生长因子滴眼液，所述的滴眼液包含蛋白保护剂、保湿剂、磷酸盐缓冲体系、渗透压调节剂、抑菌剂和抗氧化剂，其特征在于所述的蛋白保护剂选自组氨酸、吐温80和肝素钠的组合，且每100ml滴眼液包含0.1～0.2g组氨酸、0.03-0.1g吐温80和0.01～0.05g肝素钠；
所述抑菌剂和抗氧化剂分别选自三氯叔丁醇和甲硫氨酸；
或所述抑菌剂和抗氧化剂分别选自三氯叔丁醇和硫代硫酸钠；
或所述抑菌剂和抗氧化剂分别选自对羟基苯甲酸甲酯和甲硫氨酸；
每100ml滴眼液包含0.01～0.1g抑菌剂；每100ml滴眼液包含0.01～0.1g抗氧化剂；
所述磷酸盐缓冲体系的pH值为6.0-7.0；所述的渗透压调节剂选自氯化钠，每100ml滴眼液包含0.3～0.9g渗透压调节剂。
2.根据权利要求1所述的重组人酸性成纤维细胞生长因子滴眼液，其特征在于每100ml滴眼液包含1～3mg人酸性成纤维细胞生长因子。
3.根据权利要求2所述的重组人酸性成纤维细胞生长因子滴眼液，其特征在于每100ml滴眼液包含2mg人酸性成纤维细胞生长因子。
4.根据权利要求1所述的重组人酸性成纤维细胞生长因子滴眼液，其特征在于所述的保湿剂选自透明质酸钠、硫酸软骨素、聚乙烯醇、聚乙二醇、甘油、羟丙甲纤维素、右旋糖酐、羧甲基纤维素中的一种或多种，每100ml滴眼液包含0.01～1.5g保湿剂。
5.根据权利要求4所述的重组人酸性成纤维细胞生长因子滴眼液，其特征在于每100ml滴眼液包含0.1g保湿剂。
6.根据权利要求1所述的重组人酸性成纤维细胞生长因子滴眼液，其特征在于所述的缓冲盐选自磷酸盐缓冲体系，所述缓冲盐的pH值为6.0-7.0，浓度为0.01-0.05mol/L。
7.根据权利要求1所述的重组人酸性成纤维细胞生长因子滴眼液，其特征在于所述的缓冲盐的浓度为0.02mol/L。
8.根据权利要求1所述的根据权利要求1所述的重组人酸性成纤维细胞生长因子滴眼液，其特征在于所述的滴眼剂进一步包含增稠剂；所述增稠剂选自聚乙烯醇、羧甲基纤维素、甲基纤维素、羧甲基纤维素钠、聚丙烯酸钠、聚乙烯吡咯烷酮中的一种或多种，每100ml滴眼液包含0.5～2g增稠剂。
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0408146A2 (en) * | 1989-07-13 | 1991-01-16 | Merck & Co. Inc. | Method for the purification of therapeutically active recombinant acidic fibroblast growth factor |
| CN102357243A (zh) * | 2011-11-02 | 2012-02-22 | 珠海亿胜生物制药有限公司 | 重组牛碱性成纤维细胞生长因子滴眼液 |
| CN109010803A (zh) * | 2018-10-26 | 2018-12-18 | 安徽安科生物工程（集团）股份有限公司 | 一种重组人干扰素α2b滴眼液及其制备方法 |
| CN110063936A (zh) * | 2019-04-28 | 2019-07-30 | 安徽鑫华坤生物工程有限公司 | 一种重组人酸性成纤维细胞生长因子液体制剂 |
| WO2019214551A1 (zh) * | 2018-05-08 | 2019-11-14 | 华博生物医药技术（上海）有限公司 | 一种重组人血管内皮生长因子受体-抗体融合蛋白的药物组合制剂 |
| CN111315886A (zh) * | 2017-06-23 | 2020-06-19 | 珠海亿胜生物制药有限公司 | 重组人碱性成纤维细胞生长因子(rh-bFGF)以及包含rh-bFGF的药物组合物 |
| WO2021263134A1 (en) * | 2020-06-26 | 2021-12-30 | Trefoil Therapeutics, Inc. | Recombinant modified fibroblast growth factors and therapeutic uses thereof |
-
2023
- 2023-04-21 CN CN202310431075.4A patent/CN116392441B/zh active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0408146A2 (en) * | 1989-07-13 | 1991-01-16 | Merck & Co. Inc. | Method for the purification of therapeutically active recombinant acidic fibroblast growth factor |
| CN102357243A (zh) * | 2011-11-02 | 2012-02-22 | 珠海亿胜生物制药有限公司 | 重组牛碱性成纤维细胞生长因子滴眼液 |
| CN111315886A (zh) * | 2017-06-23 | 2020-06-19 | 珠海亿胜生物制药有限公司 | 重组人碱性成纤维细胞生长因子(rh-bFGF)以及包含rh-bFGF的药物组合物 |
| WO2019214551A1 (zh) * | 2018-05-08 | 2019-11-14 | 华博生物医药技术（上海）有限公司 | 一种重组人血管内皮生长因子受体-抗体融合蛋白的药物组合制剂 |
| CN109010803A (zh) * | 2018-10-26 | 2018-12-18 | 安徽安科生物工程（集团）股份有限公司 | 一种重组人干扰素α2b滴眼液及其制备方法 |
| CN110063936A (zh) * | 2019-04-28 | 2019-07-30 | 安徽鑫华坤生物工程有限公司 | 一种重组人酸性成纤维细胞生长因子液体制剂 |
| WO2021263134A1 (en) * | 2020-06-26 | 2021-12-30 | Trefoil Therapeutics, Inc. | Recombinant modified fibroblast growth factors and therapeutic uses thereof |
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