CN108309941B - 一种舍雷肽酶外用粉剂及其制备方法 - Google Patents
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Abstract
本发明公开一种舍雷肽酶外用粉剂，包括如下质量百分含量的组份：舍雷肽酶2~30%，二氧化硅颗粒9~89%，二氧化钛颗粒9~89%。本发明所述的舍雷肽酶外用粉剂，通过纳米二氧化硅和二氧化钛为载体混合吸附促伤口愈合的舍雷肽酶。二氧化硅颗粒和二氧化钛颗粒与舍雷肽酶通过酶固定化技术吸附在一起，保证了舍雷肽酶分散均匀性的同时实现对舍雷肽酶的保护防止因光照发生降解，确保酶活性基本不会降低，延长药效时间，减少了一次性用药量，降低成本和副作用发生机率，也增加了用药时促伤口愈合，配合提高抗生素抗菌的作用。
Description
技术领域
本发明属于医药技术领域，具体涉及一种二氧化硅和二氧化钛为纳米载体混合吸附的促伤口愈合的舍雷肽酶外用粉剂。
背景技术
创伤是生活中一种极为常见的伤病，比如：割伤、刺伤、挫伤等。随着人们生产活动的多样化，不仅各种意外造成的创伤的几率不断增加，创伤的程度也有很大差别，严重的创伤可引起全身反应甚至危机生命。一般创伤的出现都伴随着感染和发炎的症状，大量使用抗生素等药物虽然会暂时缓解症状，但不能解决根本的创伤修复问题，久而久之身体产生的耐药性会让病情容易复发。因此能让伤口快速愈合才能从根本上达到治疗的目标。
舍雷肽酶早期被人们发现具有蛋白溶解能力，可抑制致炎物质，多被用作化痰去脓药。同时舍雷肽酶也具有促进细胞的新生，促进伤口的愈合，与抗生素一同使用时有协同作用提高抗生素抗菌作用。所以是创伤药的良好选择。
目前市场上的舍雷肽酶大部分是片剂或者肠溶片剂，起效慢，生物利用度较低。外敷在伤口上能够大大提高舍雷肽酶的药效作用，但是暴露在空气中的舍雷肽酶很容易失活，所以普通的外用制剂在使用时易受到光线、紫外线、氧化作用等干扰，影响疗效。
发明内容
针对上述问题，本发明提出一种舍雷肽酶外用粉剂，其有效成分不易光解失活，并且载药量高，生物利用度高，药效好，安全可控。
为实现上述目的，本发明采用如下的技术方案：
一种舍雷肽酶外用粉剂，包括如下质量百分含量的组份：舍雷肽酶2~30%，二氧化硅颗粒，9~89%，二氧化钛颗粒，9~89%。
所述二氧化硅颗粒为尺寸为1~100 nm的二氧化硅纳米颗粒，优选尺寸为10~80 nm的二氧化硅纳米颗粒。
所述二氧化钛颗粒为尺寸为1~100 nm的二氧化钛纳米颗粒，优选尺寸为10~80 nm的二氧化钛纳米颗粒。
优选的，所述舍雷肽酶外用粉剂，包括如下质量百分含量的组份：舍雷肽酶10~15%，二氧化硅颗粒，35~50%，二氧化钛颗粒，35~50%。
最优的，所述舍雷肽酶外用粉剂，包括如下质量百分含量的组份：舍雷肽酶15%，二氧化硅颗粒，40%，二氧化钛颗粒，45%。
本发明第二目的在于提供一种上述舍雷肽酶外用粉剂的制备方法，包括如下步骤：
（1）将二氧化硅颗粒和二氧化钛颗粒分别加入水中搅拌形成混悬液；
（2）将舍雷肽酶加入至上述混悬液中，搅拌至酶溶解，形成酶混悬液；
（3）将步骤（2）得到的酶混悬液离心后除去上清液；
（4）将步骤（3）离心后的沉淀物，干燥成粉，即得舍雷肽酶粉剂。
进一步的，步骤（3）所述的离心在低温条件下进行，所述低温为-5~5℃。
步骤（4）所述的干燥方法有机械脱水、加热干燥、冷冻干燥、化学除湿等。
本发明利用二氧化钛的优良避光作用，防止制剂中舍雷肽酶产生光解反应，失去活性，纳米级的二氧化硅具有很大的表面积、吸附能力及化学惰性，能够提高载药量并提供相对稳定的环境。二者结合可实现舍雷肽酶体外避光应用。此外，二氧化硅可在二氧化钛表面沉积形成一层无机物包膜，降低二氧化钛的化学活性，防止其发生团聚而影响舍雷肽酶的酶活和药效的发挥，克服了单独以二氧化钛为载体对舍雷肽酶酶活的限制，使得制得的舍雷肽酶外用粉剂表现出高抗光解性，长时间保持高酶活和稳定性。
本发明通过酶固定化吸附制备出一种具有生物利用度高，药效好，安全可控等优点的舍雷肽酶外用粉剂。该舍雷肽酶外用粉剂相对于口服片剂和一般外用制剂有较大的优势。其中舍雷肽酶单位制剂用量为10~300mg/g冻干粉末。
附图说明
图1是实施例1所制备的舍雷肽酶粉剂与实施例2和实施例3所制备的粉剂酶活随时间变化图。
具体实施方式
实施例1
将15g尺寸为80nm的纳米二氧化硅颗粒和20g尺寸为10nm的纳米二氧化钛颗粒加入50ml纯水中搅拌10~30分钟，然后向混悬液中加入15g舍雷肽酶继续搅拌15~30分钟，接着于-5℃低温离心去上清，沉淀物干燥成粉，得到舍雷肽酶粉剂。
实施例2（对比实施例）
将15g尺寸为89nm的纳米二氧化硅颗粒加入50ml纯水中搅拌10~30分钟，然后向混悬液中加入15g舍雷肽酶继续搅拌15~30分钟，接着于-5℃低温离心去上清，沉淀物干燥成粉，得到舍雷肽酶粉剂。
实施例3（对比实施例）
将20g尺寸为10nm的纳米二氧化钛颗粒加入50ml纯水中搅拌10~30分钟，然后向混悬液中加入15g舍雷肽酶继续搅拌15~30分钟，接着于-5℃低温离心去上清，沉淀物干燥成粉，得到舍雷肽酶粉剂。
实施例4
配制舍雷肽酶标准浓度溶液，按梯度稀释后通过紫外分光光度计扫描后计算出浓度与紫外吸收的标准曲线方程。
将实施例1-3中的离心后的上清液进行紫外检测，计算舍雷肽酶的浓度，确认舍雷肽酶基本被纳米二氧化硅和/或纳米二氧化钛所吸附。
将实施例1-3所制备得到的舍雷肽酶粉剂放置在常温光照下，分别在0、2、4、8、12、24h测其酶活。酶活随时间变化如图1。
可以看出，24小时后，实施例1所制备的固定化后的舍雷肽酶的酶活依然保持原先的91%。
将实验用家兔经耳缘静脉麻醉。将其背部毛剪短，用10％硫化钠脱毛。常规消毒后，在其脱毛部位做2cm×2 cm面积大小的标记，用镊子将皮肤提起，用手术剪按照标记的位置和大小剪下皮肤全层，形成全层皮肤缺损的创伤模型，止血后备用，一半用作实验组，一半用作空白组。将实施例1制备的舍雷肽酶粉剂均匀洒在实验组伤口上，每天上药一次；空白组不做任何处理。伤口愈合率见表1。
表1：
| 时间/天 | 4 | 8 | 12 | 16 | 20 | 24 |
| 空白 | 10.80% | 29.35% | 43.18% | 56.45% | 68.05% | 77.42% |
| 舍雷肽酶粉剂 | 18.27% | 45.70% | 68.37% | 87.75% | 95.55% | 98.00% |
实施例5
将40g尺寸为50nm的纳米二氧化硅颗粒和45g尺寸为50nm的纳米二氧化钛颗粒加入60ml纯水中搅拌10~30分钟，然后向混悬液中加入15g舍雷肽酶继续搅拌15~30分钟，接着于0℃低温离心去上清，沉淀物干燥成粉，得到舍雷肽酶粉剂。
实施例6
将55g尺寸为10nm的纳米二氧化硅颗粒和35g尺寸为80nm的纳米二氧化钛颗粒加入60ml纯水中搅拌10~30分钟，然后向混悬液中加入10g舍雷肽酶继续搅拌15~30分钟，接着于5℃低温离心去上清，沉淀物干燥成粉，得到舍雷肽酶粉剂。
实施例7
将3.5g尺寸为1nm的纳米二氧化硅颗粒和5g尺寸为1nm的纳米二氧化钛颗粒加入40ml纯水中搅拌10~30分钟，然后向混悬液中加入1.5g舍雷肽酶继续搅拌15~30分钟，接着于2℃低温离心去上清，沉淀物干燥成粉，得到舍雷肽酶粉剂。
实施例8
将9g尺寸为30nm的纳米二氧化硅颗粒和89g尺寸为100nm的纳米二氧化钛颗粒加入80ml纯水中搅拌10~30分钟，然后向混悬液中加入2g舍雷肽酶继续搅拌15~30分钟，接着于-2℃低温离心去上清，沉淀物通过机械脱水干燥成粉，得到舍雷肽酶粉剂。
实施例9
将89g尺寸为100nm的纳米二氧化硅颗粒和9g尺寸为70nm的纳米二氧化钛颗粒加入80ml纯水中搅拌10~30分钟，然后向混悬液中加入2g舍雷肽酶继续搅拌15~30分钟，接着于-5℃低温离心去上清，沉淀物加热干燥成粉，得到舍雷肽酶粉剂。
实施例10
将50g尺寸为60nm的纳米二氧化硅颗粒和20g尺寸为40nm的纳米二氧化钛颗粒加入60ml纯水中搅拌10~30分钟，然后向混悬液中加入30g舍雷肽酶继续搅拌15~30分钟，接着于-5℃低温离心去上清，沉淀物化学祛湿干燥成粉，得到舍雷肽酶粉剂。
实施例11
将15g尺寸为6nm的纳米二氧化硅颗粒和55g尺寸为80nm的纳米二氧化钛颗粒加入60ml纯水中搅拌10~30分钟，然后向混悬液中加入8g舍雷肽酶继续搅拌15~30分钟，接着于-5℃低温离心去上清，沉淀物冷冻干燥成粉，得到舍雷肽酶粉剂。
Claims (1)
1.一种舍雷肽酶外用粉剂，其特征在于，所述的舍雷肽酶外用粉剂是由如下步骤制备得到的：
（1）将15g尺寸为80nm纳米二氧化硅颗粒和20g尺寸为10nm的纳米二氧化钛颗粒分别加入50mL纯水中搅拌10~30分钟形成混悬液；
（2）将15g舍雷肽酶加入至上述混悬液中，搅拌15~30分钟至酶溶解，形成酶混悬液；
（3）将步骤（2）得到的酶混悬液于-5℃离心后除去上清液；
（4）将步骤（3）离心后的沉淀物，干燥成粉，即得。
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