宾州大学/BioNTech全面测试冻干型mRNA-LNP稳定性-行业动态-GMP论坛

lemon 发表于 2022-5-25 09:56:04

宾州大学/BioNTech全面测试冻干型mRNA-LNP稳定性

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<p style="outline: 0px;max-width: 100%;color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;letter-spacing: 0.544px;white-space: normal;background-color: rgb(255, 255, 255);text-align: center;visibility: visible;margin-left: 0px;margin-right: 0px;box-sizing: border-box !important;overflow-wrap: break-word !important;margin-bottom: 0em;"> <span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">2022年5月4号<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">，Norbert Pardi主导，Katalin Karikó作为参与者，共同在<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">Molecular Therapy<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">发表文章：<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(0, 122, 170);">Lyophilization provides long-term stability for a lipid nanoparticle-formulated, nucleoside-modified mRNA vaccine<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">，他们对冻干后的修饰核苷酸mRNA-LNP展开稳定性测试，<span style="color: rgb(0, 0, 0);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">发现室温存储12周和4°存储24周均不会对mRNA-LNP各组分的理化性质产生显著改变。更加重要的是，在小鼠模型中，在室温存储4周或者4°存储24周的<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">冻干剂型luciferase-mRNA-LNP<span style="color: rgb(0, 0, 0);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">依旧可以高水平表达，在室温存储12周或者4°存储24周的<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">冻干剂型编码HA的流感mRNA疫苗<span style="color: rgb(0, 0, 0);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">在小鼠体内的<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">免疫原性<span style="color: rgb(0, 0, 0);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">也没有发生下降。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">这项研究通过比较不同存储温度下冻干剂型mRNA-LNP，各组分理化性质随时间发生的变化，同时测定其在体内的表达活性和免疫原性，为解决修饰核苷酸mRNA-LNP疫苗长期的存储问题奠定良好的数据基础。<h2 data-tool="mdnice编辑器" style="box-sizing: border-box;margin: 1em auto;color: rgb(119, 48, 152);font-weight: bold;line-height: 28px;font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;white-space: normal;background-color: rgb(255, 255, 255);min-height: 32px;border-bottom: 1px solid rgb(119, 48, 152);border-top-color: rgb(119, 48, 152);border-right-color: rgb(119, 48, 152);border-left-color: rgb(119, 48, 152);font-size: 22px;padding-top: 0.5em;padding-bottom: 0.5em;text-align: center;width: 552.5px;display: flex;flex-direction: column;justify-content: center;">为啥需要冻干剂型mRNA疫苗？</h2><span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">许多传统疫苗可以在2°-8°冷藏条件下仓储至少6个月，但是，目前批准的两款mRNA疫苗都需要冷链运输，这对于mRNA疫苗在不同国家和区域的运输配送造成极大的限制。在制药或者食品加工行业。冷冻干燥是一种常用的移除水分，提高产品稳定性，延长保质期的生产工艺。制备冻干剂型mRNA疫苗，解除运输中对于冷链环境的依赖，将会使得mRNA疫苗可以更加便利地触达世界上每一个需要疫苗的国家或者地区。<h2 data-tool="mdnice编辑器" style="box-sizing: border-box;margin: 1em auto;color: rgb(119, 48, 152);font-weight: bold;line-height: 28px;font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;white-space: normal;background-color: rgb(255, 255, 255);min-height: 32px;border-bottom: 1px solid rgb(119, 48, 152);border-top-color: rgb(119, 48, 152);border-right-color: rgb(119, 48, 152);border-left-color: rgb(119, 48, 152);font-size: 22px;padding-top: 0.5em;padding-bottom: 0.5em;text-align: center;width: 552.5px;display: flex;flex-direction: column;justify-content: center;">如何制备冻干剂型mRNA疫苗？</h2><span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">冻干工艺<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">，简单来说，就是把物料里面的水分给去除掉。首先将物料在低温下冻结至共晶点（<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(136, 136, 136);">物料中的水分完全冻结至冰晶时的温度<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">）以下，然后在真空环境下将冰晶升华干燥，接着再进行解吸干燥，除去剩余的结合水，从而获得干燥的产品。冷冻干燥的过程可以分为：<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">预冻，初始干燥<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">（<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(136, 136, 136);">升华干燥<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">），<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">二次干燥<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">（<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(136, 136, 136);">解吸干燥<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">）。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">在这篇文章中，研发人员首先用LNP包封luciferase-mRNA和PR8 HA-mRNA。接着，将LNP载体buffer换为 <span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">5mM PH8 Tris <span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">（<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(136, 136, 136);">10%蔗糖和10%麦芽糖<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">）。最后，将冻干mRNA-LNP存储在-80°，-20°，4°，25°，42°进行稳定性测试。<figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">Norbert Pardi制备LNP的4个组分比例</figcaption></figure><figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">mRNA-LNP冻干工艺条件</figcaption></figure><h2 data-tool="mdnice编辑器" style="box-sizing: border-box;margin: 1em auto;color: rgb(119, 48, 152);font-weight: bold;line-height: 28px;font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;white-space: normal;background-color: rgb(255, 255, 255);min-height: 32px;border-bottom: 1px solid rgb(119, 48, 152);border-top-color: rgb(119, 48, 152);border-right-color: rgb(119, 48, 152);border-left-color: rgb(119, 48, 152);font-size: 22px;padding-top: 0.5em;padding-bottom: 0.5em;text-align: center;width: 552.5px;display: flex;flex-direction: column;justify-content: center;">如何评估冻干剂型mRNA疫苗？</h2><h3 data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 1.2em;margin-bottom: 1em;color: rgb(119, 48, 152);font-weight: bold;line-height: 1.6;font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);font-size: 20px;">理化稳定性</h3><span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">将mRNA-LNP冻干处理后，正常的外形特征应该是的<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">白色的，结块状<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">。在不同稳定度下存放一定时间（<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(136, 136, 136);">4周，12周，24周<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">），冻干剂型mRNA-LNP外形依然是统一的结块结构，没有出现皱缩或者碎裂。往冻干的结块状mRNA-LNP加入水后，很快看到<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">乳白色液体，没有任何固体残渣出现<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">。<figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">存放在不同温度下的冻干mRNA-LNP外形特征和加入水重新溶解后的mRNA-LNP后的外形特征。</figcaption></figure><span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">用DLS测试冻干剂型mRNA-LNP的颗粒大小和分散度<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">。冻存在-80° 的mRNA-LNP粒径大小随着存放时间的增加并没有发生改变。在4°以下条件存放的冻干剂型mRNA-LNP在存放24周依然维持稳定的粒径。但是，存放在25°和42度的冻干剂型mRNA-LNP的<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">粒径<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(0, 0, 0);">随着存放时间不断增加<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">。存放在42°的冻干剂型mRNA-LNP在第4周时，粒径达到最大，之后一直维持不变。有意思的是，尽管粒径大小发生会发生变化，但是，存放在不同温度不同存放时间的冻干剂型mRNA的<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">分散系数<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">均小于0.10。<figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">用DLS测试冻干剂型mRNA-LNP的颗粒大小和分散度。</figcaption></figure><span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">利用Ribogreen检测冻干剂型mRNA-LNP包封效率和mRNA含量<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">，除存放在42°的mRNA-LNP外，发现在不同温度下，随着存放时间的增加，基本上没有显著的改变。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">利用毛细管电泳检测mRN完整性<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">。发现在-80°和-20°的存放24周的冻干剂型mRNA-LNP，mRNA完整性没有发生改变。而在0°以上条件下存放的mRNA-LNP，mRNA随着存放温度的增加降解会加剧。冻干剂型mRNA-LNP在4°存放24周，mRNA完整性降低10%-15%；在25°存放24周，mRNA完整性降低30%；在42°存放24周，mRNA完整性降低几乎70%。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">利用UHPLC检测<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">冻干剂型mRNA-LNP，发现不同温度下存放24周，<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">脂质体各组分<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">依然非常稳定，而且各组分比例保持不变。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">测定冻干剂型mRNA-LNP的包封效率，mRNA浓度，mRNA完整性。</figcaption></figure><span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">研发人员比较不同温度下存放4周后，<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">冻干剂型mRNA-LNP和非冻干剂型mRNA-LNP在理化性质方面的差异<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">，发现非冻干mRNA-LNP存放在0°以上的温度条件下，存放4周后，<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">粒径会增大，分散系数均会大于0.1<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">。冻干和非冻干mRNA-LNP的包封效率和mRNA浓度并未出现明显变化。冻干mRNA-LNP在mRNA完整度上要好于非冻干mRNA-LNP。总体来看，通过冻干处理mRNA-LNP，对于室温和4°存放mRNA-LNP有一定的优势。<figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">冻干剂mRNA-LNP和非冻干的湿剂mRNA-LNP不同存放条件下的稳定性比较。</figcaption></figure><h3 data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 1.2em;margin-bottom: 1em;color: rgb(119, 48, 152);font-weight: bold;line-height: 1.6;font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);font-size: 20px;">体内表达活性</h3><span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">将在不同温度下存放不同时间的冻干Luc-mRNA-LNP通过IM/ID注射到小鼠体内，观察表达的荧光强度<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">。与冻存在-80°未冻干处理的Luc-mRNA-LNP相比，冻干处理会导致Luc-mRNA表达的荧光强度发生轻微下降。将<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">在4°存贮至少24周或者在室温下存放至少4周<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">的冻干型Luc-mRNA-LNP注射到小鼠体内依然会稳定表达出荧光蛋白。存放在42°的冻干型Luc-mRNA-LNP，注射到小鼠体内表达的荧光蛋白与其它温度下存放的相比会发生明显下降，存放至24周时在小鼠体内已经检测不到荧光强度。<figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">不同温度下存放不同时间的冻干型Luc-mRNA-LNP注射到小鼠体内表达出的荧光强度。</figcaption></figure><span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">将在不同温度下存放不同时间的冻干PR8 HA-mRNA-LNP流感疫苗通过IM/ID注射到小鼠体内<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">，通过血凝素抑制试验(HAI)来评估冻干存放的流感mRNA疫苗的免疫原性变化。结果发现，冻干PR8 HA-mRNA-LNP流感疫苗在<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">室温存放12周或者4°存放至少24周<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">均不会影响其在小鼠体内的免疫原性。<figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">不同温度下存放不同时间的冻干型PR8HA-mRNA-LNP流感疫苗免疫原性的变化。</figcaption></figure><h2 data-tool="mdnice编辑器" style="box-sizing: border-box;margin: 1em auto;color: rgb(119, 48, 152);font-weight: bold;line-height: 28px;font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;white-space: normal;background-color: rgb(255, 255, 255);min-height: 32px;border-bottom: 1px solid rgb(119, 48, 152);border-top-color: rgb(119, 48, 152);border-right-color: rgb(119, 48, 152);border-left-color: rgb(119, 48, 152);font-size: 22px;padding-top: 0.5em;padding-bottom: 0.5em;text-align: center;width: 552.5px;display: flex;flex-direction: column;justify-content: center;">冻干剂型mRNA疫苗研究进展</h2><span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">Arbutus<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">开发过一款治疗 ZE-BOV siRNA-LNP，包含靶向VP24,VP35,L聚合酶蛋白的siRNA。在NPHs模型中，证实其具有完全的预防保护效力。在2014年I期临床试验中，Arbutus公司宣称<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">冻干剂型 siRNA-LNP<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">和湿剂siRNA-LNP具有相同的效果。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">2020年，董一洲等制备<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">冻干剂型luciferase-mRNA-LNP<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">，重新用水溶解后，在小鼠体内可以展现出正常的荧光强度。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">2021年，Seok-Hyun Kim制备<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">冻干剂型<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">编码Spike蛋白的<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">新冠候选liposome-mRNA疫苗<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">，在抗原设计上，使用2P（稳定融合前构象）和3Q(突变furin切割位点)。该款冻干剂疫苗在小鼠中可以触发强烈的免疫反应。<figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">Liposome-mRNA新冠疫苗</figcaption></figure><span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">2022年，AAHI开发出一款<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">冻干剂型自复制mRNA疫苗<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">，使用NLC包封samRNA-Spike，可在室温下存放6-8个月，依然具有良好的免疫原性。<figure data-tool="mdnice编辑器" style="box-sizing: border-box;margin-top: 10px;margin-bottom: 10px;color: rgba(0, 0, 0, 0.8);font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;font-size: 14px;text-align: start;white-space: normal;background-color: rgb(255, 255, 255);display: flex;flex-direction: column;justify-content: center;align-items: center;"><figcaption style="box-sizing: border-box;margin-top: 5px;text-align: center;color: rgb(136, 136, 136);">NLC/RNA混合系统安全存放时间以及冻干样品</figcaption></figure><h2 data-tool="mdnice编辑器" style="box-sizing: border-box;margin: 1em auto;color: rgb(119, 48, 152);font-weight: bold;line-height: 28px;font-family: Optima-Regular, Optima, PingFangSC-Regular, PingFangTC-Regular, PingFang SC, Cambria, Cochin, Georgia, Times, Times New Roman, serif;white-space: normal;background-color: rgb(255, 255, 255);min-height: 32px;border-bottom: 1px solid rgb(119, 48, 152);border-top-color: rgb(119, 48, 152);border-right-color: rgb(119, 48, 152);border-left-color: rgb(119, 48, 152);font-size: 22px;padding-top: 0.5em;padding-bottom: 0.5em;text-align: center;width: 552.5px;display: flex;flex-direction: column;justify-content: center;">总结</h2><span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">mRNA-LNP存放过程中触发的不稳定性主要涉及<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(0, 0, 0);">物理不稳定性和化学不稳定性<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">。<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">物理不稳定性<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">指的是纳米颗粒的聚集或者融合，表现为粒径或者分散系数的增加。<span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);color: rgb(123, 12, 0);">化学不稳定性<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">指mRNA或者脂质成分的降解。存放的外界参数，比如温度，溶解Buffer成分/ PH也可能影响到mRNA-LNP稳定性。研发人员是希望通过冻干处理，可以提升mRNA-LNP在非冷链存放时的稳定性，因此冻干工艺过程中的冻干Buffer，循环次数，温度对于mRNA-LNP维持长期的稳定性非常重要。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <span style="font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">颗粒大小<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">影响药物代谢，体内分布，药效和安全性，是非常重要的稳定性参数。冻干剂型mRNA-LNP在低于4°条件下至少可以存放24周，在室温下可以存放4-12周；在42°存放条件下，从0-4周，粒径逐渐增加到做大，从4周-24周，粒径维系不变。在这项存放条件下，最大粒径依然是小于150nm。有报道显示，在NPHs动物模型中，150nm粒径的LNP依然可以触发强烈的免疫反应。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">在这项研究中，可以看到即便在室温下存放24周，对于冻干剂型mRNA来说，包封效率和mRNA浓度基本上没有发生显著改变。尽管目前没有关于mRNA完整度限制以及疫苗有效率的mRNA完整度阈值的公开标准，但是，防止mRNA降解对于其表达活性肯定是非常重要的。mRNA降解程度与温度是呈正相关的。对于冻干剂型mRNA来说，在4°存放24周，mRNA完整度下降10-15%；在室温存放24周，mRNA完整度下降接近30%；在42°存放24周，mRNA完整性降低几乎70%。有意思的是，尽管在42度存放4周，Luc-mRNA-LNP完整性下降70%，但是，注射到体内，Luc-mRNA-LNP表达的荧光强度并没有完全丧失。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">LNP各个脂质组分影响纳米颗粒形成，稳定性以及生物学活性，因此维系各组脂质组分的稳定性对于包封药物活性来说是很重要的。在这项研究中，在所有测试条件，4个脂质组分维系其恒定的比例和完整度。值得注意的是，有一些阳离子脂质对于温度和PH是比较敏感的。<span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);"> <span style="color: rgb(34, 34, 34);font-family: -apple-system, BlinkMacSystemFont, Helvetica Neue, PingFang SC, Hiragino Sans GB, Microsoft YaHei UI, Microsoft YaHei, Arial, sans-serif;font-size: 15px;letter-spacing: 0.5px;background-color: rgb(255, 255, 255);">总的来说，这项关于冻干剂型mRNA在不同温度不同存放时间下稳定性和体内表达活性的测定数据是非常鼓舞人心的。通过冻干工艺处理后的mRNA-LNP的关键理化参数并未发生改变，而且在体内可以正常表达蛋白，具备正常的免疫原性。通过冻干工艺提升mRNA稳定性，对于下一代mRNA疫苗研发来说是非常有竞争力的优势。
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独飞侠 发表于 2022-5-25 10:19:44

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宾州大学/BioNTech全面测试冻干型mRNA-LNP稳定性