CN113198009A - 一种治疗非洲猪瘟的口服液及其制备方法 - Google Patents
一种治疗非洲猪瘟的口服液及其制备方法 Download PDFInfo
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- CN113198009A CN113198009A CN202110695500.1A CN202110695500A CN113198009A CN 113198009 A CN113198009 A CN 113198009A CN 202110695500 A CN202110695500 A CN 202110695500A CN 113198009 A CN113198009 A CN 113198009A
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- african swine
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Abstract
本发明属于口服液制备技术领域,一种治疗非洲猪瘟的口服液,其特征在于:该方剂的配方组分按重量份数计为:紫丁香40‑50份、炙麻黄15‑20份、黄岑10‑15份、知母8‑12份、当归14‑17份、连翘18‑23份、微球口服疫苗5‑8份,本发明针对猪的常规养殖环境,使用天然草本成分,对猪对人无毒无害,使用微球口服疫苗,实现了活性物质的低损失、高药物生物利用度,并大幅减增加了药物在自首部位的停留时间;原料廉价易得,制作流程简便易行;可以作为各种剂型的兽药或者饲料添加剂,减少了人力成本。
Description
技术领域
本发明属于口服液制备技术领域,特别涉及一种治疗非洲猪瘟的口服液及其制备方法。
背景技术
非洲猪瘟是通过非洲猪瘟病毒传播引发的一种只在猪类间相互感染的烈性传染病,临床症状表现为高烧、大范围的内脏器官、肠道出血和呼吸功能障碍等,具有极高的发病率和死亡率,全球主要的生猪养殖国家都曾爆发过非洲猪瘟疫情,给当地的养殖者带来沉重的经济损失。传统预防方法主要是大面积的扑杀疫情区域的动物阻断其传播途径,目前为止尚未研制出有效的疫苗对其进行治疗和控制。
草药是人类最早应用的抗病防病物质,也是最早的饲料添加剂,具有安全性高,毒副作用小,免疫增强效果好等优点。在生物安全风险剧增的今天,天然草本药物被广泛认可。紫丁香原产于我国华北地区,至今在我国已有1000多年的栽培历史,其味苦、性寒;归胃、肝、胆经。清热,解毒,利湿,退黄。主治急性泻痢,黄疸型肝炎,火眼,疮疡。已有很多研究报道紫丁香中的成分具有很强的抗病毒及止血的作用。但以紫丁香为主方的非洲猪瘟治疗口服液却未见报道。
发明内容
本发明的目的是提供一种治疗非洲猪瘟的口服液及其制备方法,具有成本低,能缓解治疗患非洲猪瘟的病猪的病情的效果。
本发明的上述技术目的是通过以下技术方案得以实现的:一种治疗非洲猪瘟的口服液,其特征在于:该方剂的配方组分按重量份数计为:紫丁香40-50份、炙麻黄15-20份、黄岑 10-15份、知母8-12份、当归14-17份、连翘18-23份、微球口服疫苗5-8份。
本发明的进一步设置为:所述微球口服疫苗通过以下步骤制备:首先将海藻酸钠加入到蛋白溶液中,采用均质匀浆机作用5-8min,使海藻酸钠完全溶解,与蛋白溶液形成均质溶液;然后将均质溶液直接喷雾于含有0.5%CaCl2的灭菌磷酸盐缓冲溶液中,搅拌15-20min,搅拌完成后转入离心机离心3-5min,转移收集液,获得微球,采用灭菌的磷酸盐缓冲溶液冲洗微球3-5次,收集微球;将上述微球分散到壳聚糖溶液中,匀浆2min后离心去上清液后,采用灭菌磷酸盐缓冲溶液清洗微球,并将清洗后的微球分散到等体积的8%的冻干保护剂中,-40℃保存24-36h,冷冻干燥得到微球口服疫苗。
本发明的进一步设置为:所述冻干保护剂含有明胶、乳糖、甘氨酸、甘露醇中的一种或两种以上。
本发明的进一步设置为:该方剂的配方组分按重量份数计优选为:紫丁香45份、炙麻黄 18份、黄岑12份、知母10份、当归13份、连翘20份、微球口服疫苗7份。
本发明的进一步设置为:配方中还包括冻干保护剂。
本发明的进一步设置为:所述冻干保护剂含有白藜芦醇、肉桂醛、槲皮素、穿心莲内酯、黄酮苷中的一种或多种
一种治疗非洲猪瘟的口服液其制备方法,其特征在于:通过以下步骤制备:
S1:将紫丁香、炙麻黄、黄岑、知母、连翘粉碎为过100-120目筛药粉,加4-8倍60-80%乙醇溶液回流提取2-3次,每次提取1.5-3h,回收乙醇至无醇味,得到紫丁香、炙麻黄、黄岑、知母、连翘提取液;
S2:将紫丁香、黄岑、知母、连翘药渣与当归混合,加10-15倍水浸泡1-2h,加热提取2-3次,每次提取1.5-2.5h,过滤后将滤液与步骤S1得到的提取液合并,过滤后进行浓缩;
S3:在步骤2得到的浓缩液中加0.02-0.04%山梨酸和微球口服疫苗,加适量蒸馏水配制成每1ml含原料药1g的药液,得到非洲猪瘟治疗口服液。。
本发明的有益效果是:。
1、本发明的以紫丁香为主方的非洲猪瘟治疗口服液,选用的都是常见原料,因此能合理控制生产成本。特别是紫丁香,在我国拥有广泛的种植,因此材料问题已经解决,且研究结果紫丁香安全无毒,以其为原料制成的非洲猪瘟治疗口服液,能提高生猪对非洲猪瘟病毒的免疫力和抵抗力,缓解病猪病情,本发明宜制作成各种剂型的兽药或者饲料添加剂,实用范围广泛。
2、本发明中采用冻干微球口服疫苗采用冻干技术,冷冻干燥后的物质,其中的营养成分损失较少,物理化学和生物性状基本不变,保存期较长,而冷冻个干燥可能会导致蛋白质变形,本发明中添加的冻干保护剂在冻干和贮存阶段保护微生物或蛋白质的生物活性,降低抗原活性的损失、变化程度、加稀释剂后能够迅速溶解,最大程度的恢复了原有的性质,能够保证微球口服疫苗的最大活性,同时保护剂还能够保护抗原免受胃内酸性环境及组织内水解酶的降解和破坏。
3、本发明中微球口服液中的海藻酸钠形成的凝胶具有较强的pH敏感性,即在酸性环境下收缩,在碱性环境下溶胀,这样,微球进入胃肠道就免受胃酸和各种蛋白酶类的影响,从而顺利到达肠部位后释药,实现靶向给药。此外,由于海藻酸钠本身的粘附性,微球进入肠道后可以黏附在肠黏膜上,在吸收部位就会长时间停留,增加了药物的生物利用度,增强免疫的保护作用。
4、本发明经临床试验证明,以紫丁香为主方的非洲猪瘟治疗口服液能明显增强生猪的病毒免疫力和抵抗力,缓解病猪病情。
5、本发明针对猪的常规养殖环境,使用天然草本成分,对猪对人无毒无害,使用微球口服疫苗,实现了活性物质的低损失、高药物生物利用度,并大幅减增加了药物在自首部位的停留时间;原料廉价易得,制作流程简便易行;可以作为各种剂型的兽药或者饲料添加剂,减少了人力成本。
具体实施方式
下面将对实施例中的技术方案进行清楚、完整地描述。显然,所描述的实施例仅仅是本发明的一部分实施例,而不是全部的实施例。基于本发明的实施例,本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1:
一种治疗非洲猪瘟的口服液其制备方法,通过以下步骤制备:
S1:将紫丁香、炙麻黄、黄岑、知母、连翘粉碎为过100目筛药粉,加4倍60%乙醇溶液回流提取3次,每次提取1.5,回收乙醇至无醇味,得到紫丁香、炙麻黄、黄岑、知母、连翘提取液;
S2:将紫丁香、黄岑、知母、连翘药渣与当归混合,加10倍水浸泡1,加热提取3次,每次提取1.5,过滤后将滤液与步骤S1得到的提取液合并,过滤后进行浓缩;
S3:在步骤2得到的浓缩液中加0.02山梨酸,加适量蒸馏水配制成每1ml含原料药1g 的药液,得到非洲猪瘟治疗口服液。
实施例2:
一种治疗非洲猪瘟的口服液其制备方法,通过以下步骤制备:
S1:将紫丁香、炙麻黄、黄岑、知母、连翘粉碎为过110目筛药粉,加5倍70%乙醇溶液回流提取3次,每次提取2h,回收乙醇至无醇味,得到紫丁香、炙麻黄、黄岑、知母、连翘提取液;
S2:将紫丁香、黄岑、知母、连翘药渣与当归混合,加12倍水浸泡2h,加热提取3次,每次提取2,过滤后将滤液与步骤S1得到的提取液合并,过滤后进行浓缩;
S3:在步骤2得到的浓缩液中加0.04%山梨酸和微球口服疫苗,加适量蒸馏水配制成每 1ml含原料药1g的药液,得到非洲猪瘟治疗口服液。
实施例3:
一种治疗非洲猪瘟的口服液其制备方法,通过以下步骤制备:
S1:将紫丁香、炙麻黄、黄岑、知母、连翘粉碎为过120目筛药粉,加8倍80%乙醇溶液回流提取2次,每次提取3h,回收乙醇至无醇味,得到紫丁香、炙麻黄、黄岑、知母、连翘提取液;
S2:将紫丁香、黄岑、知母、连翘药渣与当归混合,加15倍水浸泡2h,加热提取3次,每次提取2.5h,过滤后将滤液与步骤S1得到的提取液合并,过滤后进行浓缩;
S3:在步骤2得到的浓缩液中加0.04%山梨酸和微球口服疫苗,加适量蒸馏水配制成每 1ml含原料药1g的药液,得到非洲猪瘟治疗口服液。
将实施例1-3中所得到的的口服液利用BCA法测定4小时后pH条件下的蛋白含量,得到蛋白释放量数据如表1所示。
表1不同pH条件下微球蛋白释放量(mg)
pH=2 | pH=7.5 | pH=10 | |
实施例1 | 0 | 0 | 0 |
实施例2 | 0.04 | 0.15 | 0.25 |
实施例3 | 0.05 | 0.17 | 0.23 |
由表1可以看出,口服微球疫苗均在偏碱性条件下释放,酸性条件下几乎不释放,说明说明海藻酸钠-壳聚糖微球可以耐酸,在偏碱条件下释放,为动物口服免疫提供了依据。
将实施例2-3中的微球口服疫苗分别存放在-40℃与-20℃条件下,在1、2、3个月之后取样检测,发现微球仍为白色粉末,未变色,微球表面干燥,并无潮解现象。
将实施例1-3中的口服液给猪灌胃,在第10、15、20天后采血,同时收集粪便,采用ELISA 法分别检测血清IgG抗体和IgA抗体,得到表2数据:
表2IgG与IgA水平检测表
通过表2可以看出,在口服液中添加的微球口服疫苗能够让猪产生相应的抗体,因此,本发明形成的口服液能够刺激机体产生良好的细胞免疫应答,使机体的体液免疫保护提高。
本发明针对猪的常规养殖环境,使用天然草本成分,对猪对人无毒无害,使用微球口服疫苗,实现了活性物质的低损失、高药物生物利用度,并大幅减增加了药物在自首部位的停留时间;原料廉价易得,制作流程简便易行;可以作为各种剂型的兽药或者饲料添加剂,减少了人力成本。
Claims (7)
1.一种治疗非洲猪瘟的口服液,其特征在于:该方剂的配方组分按重量份数计为:紫丁香40-50份、炙麻黄15-20份、黄岑10-15份、知母8-12份、当归14-17份、连翘18-23份、微球口服疫苗5-8份。
2.根据权利要求1所述的一种治疗非洲猪瘟的口服液,其特征在于:所述微球口服疫苗通过以下步骤制备:首先将海藻酸钠加入到蛋白溶液中,采用均质匀浆机作用5-8min,使海藻酸钠完全溶解,与蛋白溶液形成均质溶液;然后将均质溶液直接喷雾于含有0.5%CaCl2的灭菌磷酸盐缓冲溶液中,搅拌15-20min,搅拌完成后转入离心机离心3-5min,转移收集液,获得微球,采用灭菌的磷酸盐缓冲溶液冲洗微球3-5次,收集微球;将上述微球分散到壳聚糖溶液中,匀浆2min后离心去上清液后,采用灭菌磷酸盐缓冲溶液清洗微球,并将清洗后的微球分散到等体积的8%的冻干保护剂中,-40℃保存24-36h,冷冻干燥得到微球口服疫苗。
3.根据权利要求2所述的一种治疗非洲猪瘟的口服液,其特征在于:所述冻干保护剂含有明胶、乳糖、甘氨酸、甘露醇中的一种或两种以上。
4.根据权利要求1所述的一种治疗非洲猪瘟的口服液,其特征在于:该方剂的配方组分按重量份数计优选为:紫丁香45份、炙麻黄18份、黄岑12份、知母10份、当归13份、连翘20份、微球口服疫苗7份。
5.根据权利要求1所述的一种治疗非洲猪瘟的口服液,其特征在于:配方中还包括冻干保护剂。
6.根据权利要求5所述的一种治疗非洲猪瘟的口服液,其特征在于:所述冻干保护剂含有白藜芦醇、肉桂醛、槲皮素、穿心莲内酯、黄酮苷中的一种或多种。
7.一种治疗非洲猪瘟的口服液其制备方法,其特征在于:通过以下步骤制备:
S1:将紫丁香、炙麻黄、黄岑、知母、连翘粉碎为过100-120目筛药粉,加4-8倍60-80%乙醇溶液回流提取2-3次,每次提取1.5-3h,回收乙醇至无醇味,得到紫丁香、炙麻黄、黄岑、知母、连翘提取液;
S2:将紫丁香、黄岑、知母、连翘药渣与当归混合,加10-15倍水浸泡1-2h,加热提取2-3次,每次提取1.5-2.5h,过滤后将滤液与步骤S1得到的提取液合并,过滤后进行浓缩;
S3:在步骤2得到的浓缩液中加0.02-0.04%山梨酸和微球口服疫苗,加适量蒸馏水配制成每1ml含原料药1g的药液,得到非洲猪瘟治疗口服液。
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