CN112641940A - 一种兽用灭活疫苗佐剂 - Google Patents
一种兽用灭活疫苗佐剂 Download PDFInfo
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Abstract
本发明公开了一种兽用灭活疫苗佐剂；所述疫苗佐剂采用符合《GB1886.215‑2016食品安全国家标准食品添加剂白油(又名液体石蜡)》标准的1号食品级白油；组分包含：链烷烃和环烷烃，基本不含芳烃；控制碳链区间在C14‑C26之间，碳链≤15组分的含量在5％以下,碳链在C16‑C20区间的含量为40％‑55％之间；基本不含芳烃，正构烷烃含量5‑12％。此外，所述的佐剂的链烷烃：环烷烃＝(60‑85)：(40‑15)％，优选为70％：30％；本发明的佐剂制备的兽用疫苗无明显生物刺激性，而且可提升注射部位白油佐剂的吸收，同时能够明显提高现有同等粘度产品制备疫苗的免疫效力。
Description
技术领域
本发明涉及疫苗佐剂技术领域，具体涉及一种兽用灭活疫苗佐剂，采用食品级白油，可提升现有同等粘度产品制备疫苗的注射部位吸收效果、增强免疫效力。
背景技术
兽用疫苗对于防控动物疾病，促进畜牧业发展具有举足轻重的作用。我国生物制品领域快速推进，疫苗制剂发展呈现多元化趋势，市场还是以灭活疫苗为主。灭活疫苗一般会通过制备佐剂疫苗来获得持续高效的效价。佐剂从广义上来说就是用于明显增强抗原诱导机体产生特异性免疫应答的物质，其中最常见的应用最大的佐剂物质为白油。目前白油产品主要以进口油为主，国内尚缺同等水平的白油产品。国内疫苗佐剂存在诸多安全性问题，油佐剂疫苗在动物免疫后会造成局部或全身性的应激反应，同时，佐剂的免疫效力达不到同等水平，这些都是是亟需解决的技术问题。
国内外针对白油佐剂进行充分研究；有研究发现机体对白油饱和烃吸收率随着碳数的增加而逐步降低，C26以上吸收率大幅度下降。有限的数据表明同分子量的环烷烃的吸收率与正烷烃相近，而支链烷烃的吸收率略低。烷烃代谢通过细胞色素P450系统介导的代谢通路。这个过程，正构烷烃优于异构烷烃和环烷烃，芳烃代谢会生成有生物毒性的中间产物。一般烃类的毒性以芳烃最大，而后按环烯烃，环烷烃，链烃顺序依次减小。随着烷烃分子量的增大，毒性与佐剂活性均会显著降低，C24以上基本无毒性。碳数C16-C20具有良好的活性和持续性，不同碳数烷烃混用会表现出良好的效价并且毒性基本不变。研究发现直链烷烃C6-C13会产生严重的局部炎症；C15-C20具有良好的效价，同时不存在严重副反应。C22-C24基本对机体无刺激性。
食品级白油是经过高度精炼，深度加氢除去硫、氮等杂质，重金属总量小于10ppm，无芳烃和对于碳数小于15以下的碳链严格控制的精制矿物油。经过世界粮农组织、欧洲安全局，中国等机构充分生物学论证可安全使用。在动物疫苗领域里，推广使用食品级白油作为疫苗佐剂，具有保证我国家禽产业和肉质产品健康的重要作用。
国内现有技术的白油佐剂品控要求明显低于市场对食品安全健康的需求，且用于油乳剂灭活疫苗制备时，不能完全避免免疫副反应和保持长时间较高抗体水平。因此有必要基于现有研究成果，对白油佐剂组分和碳链分布进行优化，从而获得安全、高效、稳定的食品级兽用灭活疫苗佐剂用白油
发明内容
本发明旨在针对现有注射用矿物油性能缺陷，提供一种兽用灭活疫苗佐剂，采用特别选择的食品级白油应用于兽用灭活疫苗，可以提升疫苗佐剂用白油生物体内吸收效率，而且能够改善现有注射用矿物油制备的疫苗佐剂的抗体效力。
为实现以上技术目标，本发明采用以下技术方案：
一种兽用灭活疫苗佐剂，采用符合《GB 1886.215-2016食品安全国家标准食品添加剂白油(又名液体石蜡)》标准的1号食品级白油，且所述白油40℃时的运动粘度为6.5-8mm2/s。
进一步的，所述白油的组分中碳数少于C16组分的质量百分比为0％至5％。
更进一步的，所述白油的组分中碳数大于C26组分的质量百分比为0％；
更进一步的，所述白油的组分中碳数在C16-C20区间的组分质量百分比为40％-55％；
更进一步的，所述白油的组分中正构烷烃质量百分比为5-12％。
更进一步的，所述白油的组分中链烷烃/环烷烃质量百分比为(65-80)％比(35-20)％；最好近似等于70％比30％，基本不含芳烃。
在以上技术方案中，所述组分含量测定采用气相色谱方法测定，链烷烃/环烷烃比例通过石化行业标准SH/T 0725-2002方法测得。在以上技术方案中，用于白油制备的原料均属于石化行业常规有机物或催化剂；可以利用石化炼制行业的一般技术方法获得。所述白油可依照本领域通用白油精制提炼方法制备，本发明通过化学或物理手段，分子级层面对白油组分进行针对性优化精制，优选特定组分，通过系统生物学体系评价(小白鼠白油的急性毒性测试、禽类疫苗关于安全性的吸收，关于效价的峰值和持续性测试等)，获得安全性更加良好、效价明显提升的稳定兽用灭活疫苗佐剂专用白油。
根据兽用生物制品行业中《中华人民共和国兽药典》、《兽医生物制品制造及检验规程》对白油佐剂的质量要求，对本发明白油与现有市场主流的进口竞品和样品进行了性能对比评估，以相同条件下制备的新城疫、禽流感二联灭活油乳剂疫苗为对象，考察了生物安全性、动力粘度、稳定性和效力效果。结果表明，相对于主流市场竞品及市售白油佐剂，本发明提供的食品级注射用白油具有相对更低的乳液粘度，更佳的吸收速率，无明显应激反应，更高的抗体效价，整体性能上优于现在兽用生物制品企业商业化生产使用的进口白油。
具体实施方式
以下将对本发明的具体实施方式进行详细描述。为了避免过多不必要的细节，在以下实施例中对属于公知的结构或功能将不进行详细描述。
以下实施例中所使用的近似性语言可用于定量表述，表明在不改变基本功能的情况下可允许数量有一定的变动。因此，用“大约”、“左右”等语言所修正的数值不限于该准确数值本身。在一些实施例中，“大约”表示允许其修正的数值在正负百分之十(10％)的范围内变化，比如，“大约100”表示的可以是90到110之间的任何数值。此外，在“大约第一数值到第二数值”的表述中，大约同时修正第一和第二数值两个数值。在某些情况下，近似性语言可能与测量仪器的精度有关。
除有定义外，以下实施例中所用的技术和科学术语具有与本发明所属领域技术人员普遍理解的相同含义。
以下实施例中所用的试验试剂耗材，如无特殊说明，均为常规生化试剂；所述实验方法，如无特殊说明，均为常规方法；以下实施例中的定量试验，均设置三次重复实验，结果取平均值：以下实施例中的％，如无特别说明，均为质量百分含量。
下面的例子说明了本发明中白油佐剂的特性。
在这项研究工作中，研究了各种佐剂用油的运动粘度、链烷烃/环烷烃比例、分子量，颜色等常规数据通过石化行业标准方法测得。研究了各种佐剂用油的组分，这些成分的特征是由气相色谱法测定。
测试的各种油数据如下:
表1:用于生物试验的佐剂专用白油、市场竞品白油及对照样
表中，CX-G代表本发明疫苗佐剂所专用白油；JP代表市场竞品；CX-D代表筛选过程的对照样品。
采用上述白油样品以相同条件制备新城疫、禽流感二联灭活油乳剂疫苗，具体配方如下表2：
依据《中华人民共和国兽药典》和《兽医生物制品制造及检验规程》的要求，对上述白油以相同的工艺、同批次的抗原和实验动物、在同一时间制备的新流二联灭活疫苗进行了检验评价。结果显示所制疫苗外观均为乳白色，无菌、甲醛含量、硫汞含量均通过。剂型油包水，稳定性合格。具体如下表3：
对本发明提供的专用白油(CX-GⅠ)及其他白油佐剂制备的疫苗进行生物学评价。具体情况如下：
小白鼠模型的安全性评估
取18-22g SPF级别ICR鼠，每组10只。分别注射上述白油佐剂制备的疫苗和生理盐水。ICR鼠采用腹腔注射，每只鼠注射0.5mL样品。
注射后1天，2天和3天，分别观察并记录小鼠的背毛、精神和增重情况，注射后3天，剖解查看腹腔有无病变及注射物的吸收情况。
具体情况见下表4
注：被毛分为4个等级：非常严重凌乱++++；严重凌乱+++；凌乱++；良好+；精神分为：沉郁+++；略活跃++；活跃+；剖检分为，良好，一般，较差(以腹腔腹水和脏器病变为依据)
按照该方法进行安全性评估，对结果进行综合评定。结果如表4所示，结果显示白油制备疫苗的急性毒性与白油组分中小于C16组分含量和分子量有正相关。很明显，只有组分中少于C16组分的质量百分比＜5％，分子量＞249的矿物油，才适合制备可注射的疫苗佐剂成分。
SPF鸡的安全性评估
剔除上述不可用样品后，然后评价不同白油佐剂用于新城疫和禽流感(H9亚型)灭活疫苗的安全性。取制备的疫苗，以0.5ml/只的剂量经颈背部皮下各免疫7日龄SPF鸡15只，同时设SPF空白对照组6只。于免疫后14、21和28日，从各免疫组分别随机取5只，SPF对照组随机取2只鸡作为对照。肉眼观察并记录注射部位是否出现硬结、脓肿和溃烂等现象发生；剖检观察记录注射部位的疫苗吸收情况以及是否出现硬结、脓肿和溃烂等现象；将结果记录于表5。
注：以“+”表示局部变化的严重程度。“-”表示局部无未吸收疫苗，无局部反应；“+”表示局部有极少量未吸收疫苗，基本无局部反应；“++”表示局部有较少未吸收疫苗，局部反应轻微；“+++”表示局部有极多未吸收疫苗，局部反应明显；“#”表示疫苗基本未吸收，局部有严重不良反应。
按照该方法对白油佐剂用于新城疫和禽流感(H9亚型)灭活疫苗的安全性方面关于局部反应和吸收的评估，对结果进行综合评定。结果如表5所示，结果显示白油制备疫苗的安全性(吸收和局部反应)与白油组分中＞C26组分含量和链烷烃与环烷烃的质量百分比，正构烷烃质量百分比相关。很明显，只有组分中大于C26组分的质量百分比＜0％，链烷烃与环烷烃的质量百分比为(65-85)％:(35-15)％，正构烷烃质量百分比为5-12％的矿物油，才适合制备可注射的疫苗佐剂成分。
SPF鸡的效力评估
剔除上述不可用样品后，然后评价不同白油佐剂用于新城疫和禽流感(H9亚型)灭活疫苗的效力。取上述制备的疫苗，以0.25ml/只的剂量经颈背部皮下注射7日龄SPF鸡10只，另设不免疫对照鸡5只，分别于注射后10、14、21、28和60日采集每只鸡的血液，分离血清，测定ND和H9 HI抗体效价。通过分析抗体效价，分析比较白油制备疫苗的抗体产生速度、抗体峰值。将结果记录于表6。
按照该方法对白油佐剂用于新城疫和禽流感(H9亚型)灭活疫苗的效力方面关于起效、峰值、持续性进行评估，对结果进行综合评定。结果如表6所示，结果显示白油制备疫苗的效力与白油组分中C16-C20组分含量直接相关，和各碳数组分之间偏差相关。很明显，白油的组分中碳数在C16-C20区间的组分质量百分比为40％-55％的白油，含量越高效价越好。
因此本发明最优选的配方具有如下特征：
符合《GB 1886.215-2016食品安全国家标准食品添加剂白油(又名液体石蜡)》标准的1号食品级白油，且所述白油40℃时的运动粘度为6.85mm2/s；白油的组分中碳数少于C16组分的质量百分比为2.1％；组分中碳数大于C26组分的质量百分比为0％；组分中碳数在C16-C20区间的组分质量百分比为55％；组分中正构烷烃质量百分比为12％。组分中链烷烃/环烷烃质量百分比为70比30，基本不含芳烃。本发明的佐剂制备的兽用疫苗无明显生物刺激性，而且可提升注射部位白油佐剂的吸收，同时能够明显提高现有同等粘度产品制备疫苗的免疫效力。
以上对本发明的实施例进行了详细说明，但所述内容仅为本发明的较佳实施例，并不用以限制本发明。凡在本发明的申请范围内所做的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。
Claims (7)
1.一种兽用灭活疫苗佐剂，其特征在于，采用符合《GB 1886.215-2016食品安全国家标准食品添加剂白油(又名液体石蜡)》标准的1号食品级白油，且40℃时的运动粘度为6.5-8mm2/s。
2.根据权利要求1所述的兽用灭活疫苗佐剂，其特征在于，所述的白油中碳数少于C16组分的质量百分比为0％至5％。
3.根据权利要求2所述的兽用灭活疫苗佐剂，其特征在于，所述的白油中碳数大于C26组分的质量百分比为0％。
4.根据权利要求3所述的兽用灭活疫苗佐剂，其特征在于，所述的白油中碳数在C16-C20区间的组分的质量百分比为40％-55％。
5.根据权利要求4所述的兽用灭活疫苗佐剂，其特征在于，所述的白油组分中正构烷烃质量百分比为5-12％。
6.根据权利要求5所述的兽用灭活疫苗佐剂，其特征在于，所述的白油中链烷烃与环烷烃的质量百分比为(65-85)％:(35-15)％。
7.根据权利要求5所述的兽用灭活疫苗佐剂，其特征在于，所述的白油中链烷烃与环烷烃的质量百分比为70％:30％。
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| CN115317602A (zh) * | 2022-08-31 | 2022-11-11 | 中崇信诺生物科技泰州有限公司 | 一种海诺达白油佐剂的应用及其制备的动物用灭活疫苗 |
| CN118178643A (zh) * | 2024-04-28 | 2024-06-14 | 亚培烯科技(上海)有限公司 | 饱和α烯烃低聚物作为佐剂在动物疫苗中的应用 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105396132A (zh) * | 2015-12-29 | 2016-03-16 | 天津佐沐生物科技有限公司 | 一种减轻灭活疫苗副反应的动物疫苗佐剂专用白油 |
| CN105497892A (zh) * | 2015-12-29 | 2016-04-20 | 天津佐沐生物科技有限公司 | 一种低倾点的灭活疫苗佐剂专用白油 |
| CN105535962A (zh) * | 2015-12-29 | 2016-05-04 | 天津佐沐生物科技有限公司 | 一种具有易吸收低残留功效的灭活疫苗佐剂专用白油 |
-
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- 2020-12-31 CN CN202011635264.6A patent/CN112641940A/zh active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105396132A (zh) * | 2015-12-29 | 2016-03-16 | 天津佐沐生物科技有限公司 | 一种减轻灭活疫苗副反应的动物疫苗佐剂专用白油 |
| CN105497892A (zh) * | 2015-12-29 | 2016-04-20 | 天津佐沐生物科技有限公司 | 一种低倾点的灭活疫苗佐剂专用白油 |
| CN105535962A (zh) * | 2015-12-29 | 2016-05-04 | 天津佐沐生物科技有限公司 | 一种具有易吸收低残留功效的灭活疫苗佐剂专用白油 |
Non-Patent Citations (1)
| Title |
|---|
| 中华人民共和国国家卫生和计划生育委员会: "食品安全国家标准 食品添加剂白油（又名液体石蜡）", 《中华人民共和国国家标准GB 1886.215-2016》》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115317602A (zh) * | 2022-08-31 | 2022-11-11 | 中崇信诺生物科技泰州有限公司 | 一种海诺达白油佐剂的应用及其制备的动物用灭活疫苗 |
| CN118178643A (zh) * | 2024-04-28 | 2024-06-14 | 亚培烯科技(上海)有限公司 | 饱和α烯烃低聚物作为佐剂在动物疫苗中的应用 |
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