CN106860862A - 一种抗感染性疾病的组合物和雾化喷剂及其制备方法 - Google Patents
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Abstract
本发明涉及一种制备抗感染性疾病的组合物的方法，其包括以下步骤：S1：从处于所述感染性疾病的恢复期的无传染性康复者抽取血浆；S2：从所述血浆中提取蛋白质，得到蛋白粗提物；S3：将所述蛋白粗提物冻干，得到的粉末即所述抗感染性疾病的组合物。本发明还涉及上述方法制备的抗感染性疾病的组合物，还涉及由上述组合物制备的抗感染性疾病的雾化喷剂。本发明所制备的组合物和雾化喷剂中含有针对相应的感染性疾病的抗体，可用于治疗和预防该感染性疾病。
Description
技术领域
本发明涉及感染性疾病治疗领域，更特别地，涉及一种抗感染性疾病的组合物和雾化喷剂及其制备方法。
背景技术
被动免疫疗法是把已经获得免疫性的动物的血清注射到未经免疫的机体所产生的短时期的免疫,机体中的抗体不是自己产生而是从外界获得的。人工被动免疫是注射含有特异性抗体的免疫血清或纯化免疫球蛋白抗体，或注射细胞因子等细胞免疫制剂，使机体即刻获得特异性抗感染免疫能力，及时发挥作用。
采集恢复期患者血浆/血清/全血，用于治疗危重患者属于被动免疫，在多种疾病上得到应用，如以呼吸道传播为主的急性传染病严重急性呼吸综合征(SARS)疫情蔓延期间，香港及亚洲多地区医院报道使用从恢复期SARS患者采集的血浆，成功治疗SARS重症患者。埃博拉病毒性疾病(EVD)爆发流行期间，从恢复期患者采集全血给少数EVD患者治疗取得了希望性进展，有报道称在应用全血或血浆治疗EVD可降低10％的死亡率。
然被动免疫疗法一般都是采用静脉输注的方式进行，这也是此方法的局限性，输注前要进行血液配型，限制了此方法的推广。如果能够通过其他途径给药，将促进被动免疫疗法的推广。
雾化喷剂是将生物有效分子雾化后通过口腔经咽、喉、气管、到肺泡，主要由肺泡吸收，人体的呼吸系统主要由鼻、咽、喉、气管、支气管、细支气管、终末细支气管、呼吸细支气管、肺泡管及肺泡囊组成。从气管到肺泡，气道的管径越来越小，但数量却大大增加。呼吸道表面覆盖假复层纤毛柱状上皮，纤毛细胞间夹有杯状细胞，柱状细胞游离面可见纤毛。
生物有效分子在肺部主要的吸收部位是肺泡。正常成人大约有几亿个肺泡，总表面积约为200m2，与小肠粘膜的微绒毛总面积大致相当。肺泡是半球状的囊泡，由单层扁平上皮细胞构成，厚度仅0.1-0.5μm，相邻肺泡之间的薄层结缔组织为肺泡隔。肺泡隔内有丰富的毛细血管及胶原纤维和弹性纤维。肺泡壁和毛细血管壁间的厚度仅约1μm，是气体交换和生物有效分子吸收的良好场所。由于肺泡吸收表面积大、毛细血管网丰富、转运距离极小，因此肺部给药吸收迅速。另外，肺部吸收相对胃肠道吸收来说，对生物有效分子的代谢作用小，可避免肝脏的首过效应影响。对于静脉回输，或者肌注来说，通过雾化经肺部吸收可避免输血过程的配型限制，扩大使用范围。
发明内容
为解决以上问题，本发明提供了一种制备抗感染性疾病的组合物的方法，其包括以下步骤：
S1：从处于所述感染性疾病的恢复期的无传染性康复者抽取血浆；
S2：从所述血浆中提取蛋白质，得到蛋白粗提物；
S3：将所述蛋白粗提物冻干，得到的粉末即所述抗感染性疾病的组合物。
优选地，所述感染性疾病例如非呼吸道传播的HIV、HP等等导致的感染性疾病，以及经过呼吸道传播的流感病毒，冠状病毒，赛卡病毒，禽流感病毒等等导致的感染性疾病。
优选地，S2中通过饱和硫酸铵沉淀法从所述血浆中提取蛋白质。
优选地，S3中，在冻干之前，向所述蛋白粗提物中加入蛋白保护剂，然后再进行冻干。
优选地，所述蛋白保护剂是抗坏血酸，工作浓度为0.03mg/ml。
本发明还提供了一种抗感染性疾病的组合物，其由上述方法制备得到。
本发明还提供了一种抗感染性疾病的雾化喷剂，其通过对上述抗感染性疾病的组合物进行杀灭细菌和病毒处理后溶解于药学可接受的溶剂中，得到所述通用肿瘤疫苗雾化喷剂。
优选地，所述药学可接受的溶剂为无菌水。
优选地，通过使用钴60照射来杀灭所述抗感染性疾病的组合物中的细菌和病毒。
从恢复期患者血浆中提取的蛋白含有相关疾病的抗体，以及可直接用于感染患者的治疗，及未感染患者的预防保健。
通过雾化途径经呼吸道至肺部，喷剂通过呼吸道由口腔、咽、喉、气管、支气管、肺部等部分完成吸收过程。主要经过口腔黏膜及肺部吸收，人口腔的舌下粘膜上皮未被角质化，最有利于生物有效分子的全身吸收；其次是齿龈和硬腭，一般来说，人口腔粘膜对生物分子的通透性：舌下粘膜>颊粘膜>牙龈。口腔粘膜吸收没有肝脏的首过效应，有效成分直接由颈内丰富的静脉进入循环系统，迅速发挥作用。起效所需时间短。对生物有效分子的依从性好，吸收过程和吸收效果易于控制；粘膜不易损伤，易于修复；避免胃肠道的消化酶的消化作用；可发挥局部或全身作用。
肺部具有巨大的表面积，毛细血管丰富，能够快速高效的吸收分子量较大的蛋白多肽类物质，有效吸收率高，肺泡囊壁由单层上皮细胞构成，这些细胞紧靠毛细血管网，气血屏障较小，蛋白和多肽类通过空气血液途径交换的距离短，速度快；肺部酶分布集中而且生物活性低，蛋白质和多肽类物质通过肺部吸收仍可保持生物活性；肺部吸收生物有效分子可以避免胃肠道对生物有效分子的不良影响及能避开肝脏的首关效应，提高生物利用度。对于静脉回输，或者肌注来说，通过雾化经肺部吸收可避免输血过程的配型限制，扩大使用范围。
具体实施方式
以下对本发明的原理和特征进行描述，所举实例只用于解释本发明，并非用于限定本发明的范围。
实施例1.抗H1N1型流感的雾化喷剂
本实施例中，通过以下方法来制备抗H1N1型流感的雾化喷剂：
1)从患H1N1型流感康复后六个月内的康复者体内提取血浆；
2)取血浆，用生理盐水稀释，边搅拌边缓慢逐滴加入饱和硫酸铵溶液至硫酸铵终浓度50％,4℃静置6h或过夜,4000r/min离心15-30min,分离沉淀，将沉淀溶于生理盐水,超滤除盐浓缩，得到包含针对H1N1型流感病毒的抗体的蛋白粗提物；
3)在蛋白粗提物中加入0.03mg/ml抗坏血酸作为蛋白保护剂，而后制成冻干粉；
4)10℃下经钴60照射进行病毒灭活处理，照射剂量为15-30kGy，将灭活后的冻干粉分装保存；
5)用双蒸水溶解分装后的冻干粉，制成喷剂溶液，用雾化喷剂装置进行雾化吸收。
所得的雾化喷剂通过蛋白免疫印迹检测证实，里面确实含有针对H1N1型流感病毒的抗体。
实施例2.抗HPV感染的雾化喷剂
本实施例中，通过以下方法来制备抗HPV感染的雾化喷剂：
1)从患HPV感染康复后六个月内的康复者体内提取血浆；
2)取血浆，用生理盐水稀释，边搅拌边缓慢逐滴加入饱和硫酸铵溶液至硫酸铵终浓度50％,4℃静置6h或过夜,4000r/min离心15-30min,分离沉淀，将沉淀溶于生理盐水,超滤除盐浓缩，得到包含针对HPV的抗体的蛋白粗提物；
3)在蛋白粗提物中加入0.03mg/ml抗坏血酸作为蛋白保护剂，而后制成冻干粉；
4)10℃下经钴60照射进行病毒灭活处理，照射剂量为15-30kGy，将灭活后的冻干粉分装保存；
5)用双蒸水溶解分装后的冻干粉，制成喷剂溶液，用雾化喷剂装置进行雾化吸收。
所得的雾化喷剂通过蛋白免疫印迹检测证实，里面确实含有针对HPV的抗体。
实施例3.抗禽流感病毒感染的雾化喷剂
本实施例中，通过以下方法来制备抗禽流感病毒感染的雾化喷剂：
1)从患禽流感病毒感染康复后六个月内的康复者体内提取血浆；
2)取血浆，用生理盐水稀释，边搅拌边缓慢逐滴加入饱和硫酸铵溶液至硫酸铵终浓度50％,4℃静置6h或过夜,4000r/min离心15-30min,分离沉淀，将沉淀溶于生理盐水,超滤除盐浓缩，得到包含针对禽流感病毒的抗体的蛋白粗提物；
3)在蛋白粗提物中加入0.03mg/ml抗坏血酸作为蛋白保护剂，而后制成冻干粉；
4)10℃下经钴60照射进行病毒灭活处理，照射剂量为15-30kGy，将灭活后的冻干粉分装保存；
5)用双蒸水溶解分装后的冻干粉，制成喷剂溶液，用雾化喷剂装置进行雾化吸收。
所得的雾化喷剂通过蛋白免疫印迹检测证实，里面确实含有针对禽流感病毒的抗体。
以上所述仅为本发明的较佳实施例，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (9)
1.一种制备抗感染性疾病的组合物的方法，其特征在于，包括以下步骤：
S1：从处于所述感染性疾病的恢复期的无传染性康复者抽取血浆；
S2：从所述血浆中提取蛋白质，得到蛋白粗提物；
S3：将所述蛋白粗提物冻干，得到的粉末即所述抗感染性疾病的组合物。
2.根据权利要求1所述的方法，其特征在于，所述感染性疾病包括以下病原体导致的感染性疾病：HIV、HP、流感病毒、冠状病毒、赛卡病、禽流感病毒。
3.根据权利要求1所述的方法，其特征在于，S2中通过饱和硫酸铵沉淀法从所述血浆中提取蛋白质。
4.根据权利要求1-3中任一项所述的方法，其特征在于，S3中，在冻干之前，向所述蛋白粗提物中加入蛋白保护剂，然后再进行冻干。
5.根据权利要求4所述的方法，其特征在于，所述蛋白保护剂是抗坏血酸。
6.一种抗感染性疾病的组合物，其特征在于，由权利要求1-5中任一项所述的方法制备得到。
7.一种抗感染性疾病的雾化喷剂，其特征在于，通过对权利要求6所述的通用肿瘤疫苗组合物进行杀灭细菌和病毒处理后溶解于药学可接受的溶剂中，得到所述通用肿瘤疫苗雾化喷剂。
8.根据权利要求7所述的雾化喷剂，其特征在于，所述药学可接受的溶剂为无菌水。
9.根据权利要求7或8所述的雾化喷剂，其特征在于，通过使用钴60照射来杀灭所述抗感染性疾病的组合物中的细菌和病毒。
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