Ö¬ÖÊÄÉÃ׿ÅÁ£×÷Ϊ COVID-19 mRNA ÒßÃçµÄÖØÒª×é³É²¿·Ö£¬ÔÚÓÐЧ±£»¤ mRNA ºÍ½« mRNA תÔ˵½Ï¸°ûµÄ¹ý³ÌÖз¢»Ó¹Ø¼ü×÷Óá£ÂãµÄ DNA »ò RNA ÔÚÌåÒºÖÐÈÝÒ×±»ºËËáøѸËÙ½µ½â£¬ÄÑÒÔÔÚ°Ð×éÖ¯ÖлýÀÛ¡£ÇÒÃâÒßϵͳҲÄܹ»Ê¶±ð²¢½µ½âÍâÔ´ÐԵĺËËáÒý·¢ÃâÒß·´Ó¦[1]¡£
»ùÓÚ DNA »ò RNA µÄ»ùÒòÁÆ·¨Óöµ½µÄ×î´óÎÊÌâ±ãÊÇÒ©ÎïµÝËÍ¡£ÎªÁËʵÏÖ°²È«ÓÐЧµÄºËËáµÝËÍ£¬¿Æѧ¼ÒÃÇ¿ª·¢ÁËÖ¬ÖÊÄÉÃ׿ÅÁ££¨Lipid nanoparticle, LNP£©ÒÔ±£»¤ºËËá²»±»½µ½â£¬×î´óÏ޶ȵØÏò°Ðϸ°ûµÝËÍ£¬²¢¼õÉÙºËËá¶ÔÍÑ°Ðϸ°ûµÄ±©Â¶¡£
¡°Ö¬ÖÊÌ塱һ´Ê³öÏÖÓÚÉÏÊÀ¼ÍÁùÊ®Äê´ú£¬µ±Ê±·¢ÏÖ·â±ÕµÄÖ¬ÖÊË«²ãÄÒÅÝÄÜÔÚË®ÖÐ×Ô·¢Ðγɡ£
Ö¬ÖÊÌåÊÇÒ»ÖÖÄÉÃ×ÔØÌ壬ÓÉÒ»¸ö»ò¼¸¸öÖ¬ÖÊË«²ã×é³É£¬´óСÔÚ 20 nM µ½ 1000 nM Ö®¼ä£¬Ç×Ë®ÐÔÒ©Îï¿ÉÒÔ·â±ÕÔÚÖ¬ÖÊÌåµÄË®ÐÔÄÚ²¿ÇøÓò£¬¶øÊèË®ÐÔÒ©Îï¿ÉÒÔ°ü¹üÔÚ֬˫²ãµÄÌþÁ´ÇøÓòÖÐ[1]¡£Òò´Ë£¬Ö¬ÖÊÌå±»¹ã·ºÑо¿ÓÃÓÚÒ©ÎïÊäËÍ¡£
Ö¬ÖÊÌåÄܹ»Îȶ¨ÓÃÓÚÖÎÁƵĻ¯ºÏÎï²¢¿Ë·þϸ°ûºÍ×éÖ¯ÎüÊÕµÄÕÏ°[2][3]£¬¸ÄÉÆ»¯ºÏÎï¶Ô¼²²¡²¿Î»µÄ°ÐÏòÐÔ£¬´Ó¶ø¼õÉÙÔÚ·Ç°ÐÆ÷¹ÙÖеĻýÀÛ[4]¡£¸ù¾Ý¸øҩ;¾¶ºÍ¼²²¡²¿Î»£¬½áºÏ²»Í¬µÝËÍƽ̨µÄÖ¬ÖÊÌå¿ÉÒÔ½øÒ»²½¸ÄÉÆ·â×°»¯ºÏÎïµÄµÝËÍ¡£
ͼ 1. ²»Í¬ÀàÐÍÖ¬ÖÊÌåÒ©ÎïµÝËÍϵͳµÄʾÒâͼ[4]¡£
(A) ´«Í³Ö¬ÖÊÌ壻(B) ¾ÛÒÒ¶þ´¼»¯Ö¬ÖÊÌ壻(C) ÅäÌå°ÐÏòÖ¬ÖÊÌ壻(D) ÖÎÁÆÕï¶ÏÖ¬ÖÊÌå¡£
Ö¬ÖÊÄÉÃ׿ÅÁ£ÊÇÓÉÒ»¸ö (µ¥²ã) »ò¶à¸ö (¶à²ã) Á×֬˫²ã×é³ÉµÄÇòÐÎÄÒÅÝ£¬Í¨³£ÓÉËÄÖֳɷÖ×é³É£ºÑôÀë×ÓÖ¬ÖÊ (Cationic lipids)¡¢¸¨ÖúÖ¬ÖÊ (Helper lipids)¡¢µ¨¹Ì´¼ (Cholesterol) ºÍ¾ÛÒÒ¶þ´¼»¯Ö¬ÖÊ (Pegylated lipids, PEG-lipids) (ͼ 2)¡£
ͼ 2. º¬ÓÐ mRNA µÄÖ¬ÖÊÄÉÃ׿ÅÁ£µÄʾÒâͼ[1]¡£
▐ ÑôÀë×ÓÖ¬ÖÊ
×îÔçµÄÖ¬ÖÊÄÉÃ׿ÅÁ£Ê¹ÓõÄÊÇÑôÀë×ÓÖ¬ÖÊ£¬ÆäÈÝÒ×Óë´ø¸ºµçºÉµÄºËËá½áºÏ¡£µ«ÊÇÕâÖÖ»ùÓÚÑôÀë×ÓÖ¬ÖʵĵÝËÍϵͳÔÚÌåÄÚºÍÌåÍⶼ¾ßÓж¾ÐÔºÍÃâÒßÔÐÔ¡£ÀýÈç DOTAP ºÍ DOTMA ¿ÉÒÔ±»´ø¸ºµçºÉµÄѪÇåµ°°×Öкͣ¬µ¼Ö¶¾ÐԺ͹¦Ð§½µµÍ[1]¡£
×÷ΪһÖÖÌæ´ú·½°¸£¬¿ÉµçÀëµÄÑôÀë×ÓÖ¬ÖÊ£¨Ionizable cationic lipids£©±»¿ª·¢³öÀ´£¬Ëü¼õÉÙÁËÑôÀë×ÓÖ¬ÖÊÄÉÃ׿ÅÁ£ÒýÆðµÄ¶¾ÐÔ£¬Í¬Ê±±£ÁôÁËÓÐЧµÄתȾÌØÐÔ¡£¿ÉµçÀëµÄÑôÀë×ÓÖ¬ÖÊ£¨Èç DLin-KC2-DMA ºÍ D-Lin-MC3-DMA£©µÄ¿ÉµçÀëÊå°·²¿·ÖÔÚËáÐÔ pH ֵϴø¾»ÕýµçºÉ£¬µ«ÔÚѪҺѻ·ÏµÍ³ÖÐ (pH 7.4) ±£³ÖÖÐÐÔ[5]¡£ÕâÖÖ pH Ãô¸ÐÐÔÓÐÀûÓÚºËËáÒ©ÎïÔÚÌåÄڵĴ«µÝ£¬ÒòΪ±£³ÖÖÐÐÔµÄÖ¬ÖÊÄÉÃ׿ÅÁ£ÓëѪϸ°ûÒõÀë×ÓĤµÄÏ໥×÷ÓýÏÉÙ£¬´Ó¶øÌá¸ßÁËÖ¬ÖÊÄÉÃ׿ÅÁ£µÄÉúÎïÏàÈÝÐÔ¡£
Ò»°ãÇé¿öÏ£¬Ï¸°ûͨ¹ýÄÚÍÌ×÷ÓÃÉãÈ¡Ö¬ÖÊÄÉÃ׿ÅÁ£¡£µ±Ö¬ÖÊÄÉÃ׿ÅÁ£½øÈë pH Öµ½ÏµÍµÄÄÚºÌåÖУ¬¿ÉµçÀëµÄÑôÀë×ÓÖ¬Öʱ»ÖÊ×Ó»¯±äµÃ´øÕýµçºÉ£¬Õâ´Ù½øÄÚºÌåĤµÄ²»Îȶ¨£¬Ê¹Ä¤ÆÆÁÑ£¬Ö¬ÖÊÄÉÃ׿ÅÁ£·¢ÉúÄÚºÌåÌÓÒÝ[6]¡£Ä¿Ç°£¬ÓÐÎåÖÖÖ÷ÒªµÄ¿ÉµçÀëÖ¬ÖÊÀàÐ͹㷺ÓÃÓÚ RNA µÝËÍ[7]¡£
ͼ 3. ¿ÉµçÀëÖ¬ÖʺÍÎåÖÖÖ÷Òª½á¹¹Àà±ðµÄ¿ÉµçÀëÖ¬ÖÊÆÆ»µÄÚÌåµÄ»úÖÆ[7]¡£
ÒõÀë×ÓÄÚÌåÁ×Ö¬ºÍÖÊ×Ó»¯µÄ¿ÉµçÀëÖ¬ÖÊÐγɵÄ׶ÐÎÀë×Ó¶Ô¿ÉÒÔÆÆ»µË«²ã½á¹¹£¬´Ù½øÄÚÌåÌÓÒÝ¡£¸ù¾ÝÆä½á¹¹ÌØÐÔ£¬RNA µÝË͵ĿɵçÀëÖ¬ÖÊ¿É·ÖΪ²»±¥ºÍ£¨º¬Óв»±¥ºÍ¼ü£©¡¢¶à⣨º¬ÓÐÁ½¸öÒÔÉÏ⣩¡¢¾ÛºÏ£¨º¬ÓоۺÏÎï»òÊ÷֦״¾ÛºÏÎ¡¢¿ÉÉúÎï½µ½â£¨º¬ÓпÉÉúÎï½µ½â¼ü£©ºÍβ²¿Ö§Á´Ö¬ÖÊ£¨°üº¬·Ö֧β²¿£©¡£
▐ ¾ÛÒÒ¶þ´¼»¯Ö¬ÖÊ
¾ÛÒÒ¶þ´¼»¯Ö¬ÖÊÊÇÁíÒ»ÖÖÖØÒªµÄÖ¬ÖÊÄÉÃ׿ÅÁ£³É·Ö£¬¾ÛÒÒ¶þ´¼Ö¬ÖÊ£¨Èç DMG-PEG 2000 ºÍ DSPE-MPEG-2000£©ÊÇÓÉÇ×Ë®ÐÔ¾ÛÒÒ¶þ´¼Í¨¹ýÁ×ËáÑΡ¢¸ÊÓÍ»òÆäËûÁ¬½ÓÎïÓëÊèË®ÐÔÍé»ùÁ´½áºÏ¶ø³É¡£ ¾ÛÒÒ¶þ´¼»¯Ö¬ÖÊλÓÚÖ¬ÖÊÄÉÃ׿ÅÁ£µÄ±íÃ棬֬ÖʽṹÓòÉîÂñÔÚ¿ÅÁ£ÖУ¬¾ÛÒÒ¶þ´¼½á¹¹Óò´Ó±íÃæÉì³ö¡£
¾ÛÒÒ¶þ´¼»¯Ö¬ÖÊ¿ÉÒÔÑÓ³¤Ö¬ÖÊÌåµÄÑ»·Ê±¼ä£¬ÒòΪ¾ÛÒÒ¶þ´¼ÐγÉÁËÒ»¸ö¿Õ¼äÆÁÕÏ£¬·ÀֹѪ½¬µ°°×µÄ½áºÏ£¬·ñÔò»áµ¼ÖÂËüÃDZ»Íø×´ÄÚƤϸ°û¿ìËÙÇå³ý[8]¡£
´ËÍ⣬¾ÛÒÒ¶þ´¼»¯Ö¬ÖÊ»¹¿ÉÒÔ¿ØÖÆÄÉÃ׿ÅÁ£µÄ´óС¡£ÕâÊÇÒòΪÔÚÖÆÔì¹ý³ÌÖÐµÍ pH ºÍÒÒ´¼»·¾³½«´Ù½øÖ¬ÖÊÄÉÃ׿ÅÁ£¾Û¼¯Èںϣ¬¶ø¾ÛÒÒ¶þ´¼»¯Ö¬ÖʵÄλ×èÆÁÕÏ·ÀÖ¹ÁËÕâÖÖÇé¿öµÄ·¢Éú£¬²¢ÓÐÖúÓÚ²úÉú¾ßÓÐÕ¶à·ÖÉ¢ÐÔºÍСÁ£¾¶µÄ¾ùÔÈ¿ÅÁ£Èº (ͨ³£Îª50¨C100 nM)[8][9]¡£ÍêȫȱÉÙ¾ÛÒÒ¶þ´¼Ö¬ÖʵÄÅä·½»á²úÉú²»Îȶ¨µÄ¡¢¶à·ÖÉ¢µÄÖ¬ÖÊÄÉÃ׿ÅÁ£[9]¡£
ͼ 4. ¾ÛÒÒ¶þ´¼Ö¬Öʺ¬Á¿¶ÔÖ¬ÖÊÄÉÃ׿ÅÁ£ÐÎ̬ºÍ´óСµÄÓ°Ïì[10]¡£
▐ µ¨¹Ì´¼
µ¨¹Ì´¼¾ßÓÐÊèË®ÐԺ͸ÕÐÔ£¬¿ÉÌî³äÖ¬ÖÊÌåĤÄÚÖ¬ÖÊÖ®¼äµÄ¿Õ϶£¬´Ù½øÄÒÅݵÄÎȶ¨ÐÔ¡£µ¨¹Ì´¼ÑÜÉúÎïµÄ·Ö×Ó¼¸ºÎ½á¹¹¿ÉÒÔ½øÒ»²½Ó°ÏìÖ¬ÖÊÄÉÃ׿ÅÁ£µÄµÝËÍЧ¹ûºÍÉúÎï·Ö²¼¡£¸¨ÖúÖ¬ÖʶàΪÁ×Ö¬£¨phospholipids)£¬Èç DSPC ºÍ DOPE£¬Í¨¹ý´Ù½øÓëϸ°ûºÍÄÚÌåĤµÄÈںϣ¬´Ù½øϸ°ûÉãÈ¡ºÍÄÚºÌåÊÍ·Å¡£
ÒÑÖª³¬¹ý 40% ÖÎÁÆ°©Ö¢µÄС·Ö×ÓÒ©ÎïÔÚË®ÖÐÈܽâ¶ÈµÍ£¬¶øÖ¬ÖÊÌå×÷ΪҩÎïµÝËÍϵͳÄܹ»·â×°ÕâЩҩÎï²¢Ìá¸ßÆäË®ÈÜÐÔ£¬½µµÍÁËÒ©Îï¶ÔÕý³£×éÖ¯µÄ¶¾ÐÔ£¬ÑÓ³¤ÁËÒ©ÎïµÄÍ£Áôʱ¼ä¡£Ðí¶àÖ¬ÖÊÄÉÃ׿ÅÁ£Ò©ÎïÖƼÁÒѾ±»¹ã·ºÓ¦ÓÃÓÚÐí¶àÁÙ´²ÊÔÑ飬×÷Ϊ¿¹°©¡¢¿¹Ñס¢¿¹ÉúËØ¡¢¿¹Õæ¾ú¡¢Âé×í¼ÁºÍÆäËûÒ©ÎïºÍ»ùÒòÁÆ·¨µÄµÝËÍϵͳ£¬ÓÈÆäÊÇÔÚµÝËͺËËáÒ©ÎïÁìÓò¡£
ÀýÈ磬×îÔç»ñÅúµÄÖ¬ÖÊÌåÒ©Îï Doxil£¬Ò»ÖÖ¿¹Ö×ÁöÒ©ÎﰢùËصÄÖ¬ÖÊÄÉÃ×Á£ÖƼÁ£¬Ê¹ÓÃÄÉÃ׿ÅÁ£ÑÓ³¤ÔÚÈËѪ½¬ÖеÄÑ»·Ê±¼ä£¬Í¬Ê±½µµÍ°¢Ã¹ËصÄÐÄÔ඾ÐÔ[11]¡£
Æä´Î£¬ºËËáÒ©Îï Patisiran£¬Ò»ÖÖÔÚÖ¬ÖÊÄÉÃ׿ÅÁ£¸ºÔØµÄ siRNA Ò©Î¿É¼õÉÙ¸ÎÔàÖÐת¼××´ÏÙËص°°×µÄÐγɣ¬×î½ü»ñµÃ FDA Åú×¼ÓÃÓÚÖÎÁÆÒÅ´«ÐÔת¼××´ÏÙËص°°×½éµ¼µÄµí·ÛÑù±äÐÔ¡£ËüÊÇ×îÔç»ñÅúÖ¬ÖÊÄÉÃ׿ÅÁ£ÖƼÁºËËáÒ©Î±»ÊÓ×÷ºËËáÁÆ·¨·¢Õ¹µÄÖØÒªÀï³Ì±®¡£
´ËÍ⣬֬ÖÊÄÉÃ׿ÅÁ£µÄ³É¹¦Ó¦ÓÃÊÇ Pfizer/BioNTech ºÍ Moderna »ñÅúÉÏÊеÄÁ½ÖÖ COVID-19 mRNA ÒßÃçµÄµÝËÍÔØÌå¡£Á½ÖÖ mRNA ÒßÃçµÄÖ¬ÖÊÄÉÃ׿ÅÁ£µÄ×é³É·Ç³£ÏàËÆ£¨Í¼ 5A£©¡£
ͼ 5. COVID-19 mRNA ÒßÃçÖ¬ÖÊÄÉÃ׿ÅÁ£[11]¡£
A£ºCOVID-19 mRNA ÒßÃçÖ¬ÖÊÄÉÃ׿ÅÁ£µÄÖ¬Öʳɣ»B£ºCOVID-19 mRNA ÒßÃçÖ¬ÖÊÄÉÃ׿ÅÁ£µÄÖ¬ÖʳɷֵĽṹ¡£
²»¹ý£¬Î´¾ÐÞÊεÄÖ¬ÖÊÄÉÃ׿ÅÁ£Í¬Ñù¾ßÓоÖÏÞÐÔ¡£Ö¬ÖÊÄÉÃ׿ÅÁ£»áÔÚ¸ÎÔà»ý¾Û£¬È±·¦¸ÎÔàÒÔÍâÆ÷¹Ù°ÐÏòÑ¡ÔñÐÔ¡£
һƪÌâΪ"Lung-selective mRNA delivery of synthetic lipid nanoparticles for the treatment of pulmonary lymphangioleiomyomatosis" µÄÎÄÕÂÖУ¬Ñо¿ÍŶÓͨ¹ýÎÄ¿âɸѡ·¢ÏÖ N ϵÁÐÖ¬ÖÊ£¨Î²²¿º¬ÓÐõ£°·¼ü£©ÖƱ¸µÄÄÉÃ׿ÅÁ£Äܹ»Ñ¡ÔñÐԵؽ« Cre mRNA µÝË͵½Ð¡Êó·ÎÖУ¬¶øÇÒÖ»Òªµ÷Õû N ϵÁÐÖ¬ÖʵÄÍ·²¿½á¹¹¾Í¿ÉÒÔʵÏÖ°ÐÏò²»Í¬µÄ·Îϸ°ûÀàÐÍ£¨Í¼ 4£©¡£
ÔÚ 306-N16B LNP ÖУ¬33.6% µÄ·ÎÄÚƤϸ°û±»×ªÈ¾£¬1.5% µÄÉÏƤϸ°ûºÍ 1.9% µÄ¾ÞÊÉϸ°û±»×ªÈ¾¡£¶ø 113-N16B LNP ÓÅÏȽ« Cre mRNA ´«µÝ¸øÄÚƤϸ°û£¨Õ¼ÄÚƤϸ°û×ÜÊýµÄ 69.6%£©£¬µ«Ò²´«µÝ¸ø¾ÞÊÉϸ°û (18.9%) ºÍÉÏƤϸ°û (7.3%) ¡£ÕâÒ»Ñо¿ÓÐÖúÓÚ½â¾öÖ¬ÖÊÄÉÃ׿ÅÁ£Ïò¸ÎÔàÒÔÍâÆ÷¹Ù£¨Èç·ÎºÍÉö£©µÄÓÐЧµÝË͵ÄÎÊÌâ¡£
ͼ 6. ͨ¹ýµ÷ÕûÖ¬ÖʵÄÍ·²¿½á¹¹µÄÍ·²¿½á¹¹£¬Ö¬ÖÊÄÉÃ׿ÅÁ£¿ÉÒÔ°ÐÏò²»Í¬ÀàÐ͵ķÎϸ°û[12]¡£
(A) ½« Cre mRNA ´«µÝµ½ tdTomato ת»ùÒò Ai14 СÊ󸹲¿µÄʾÒâͼ¡£(B, C) IVIS ³ÉÏñϵͳÅÄÉãµÄ Ai14 СÊó²»Í¬Æ÷¹ÙÖÐ tdTomato Ó«¹âµÄͼÏñ£¬¹²¾Û½¹ÏÔ΢¾µÅÄÉãµÄ·Î×éÖ¯µÄÃâÒßÓ«¹âͼÏñ£¬ÒÔ¼°Í¨¹ýÁ÷ʽϸ°ûÊõ¶Ô 306-N16B (B) ºÍ 113-N16B (C) LNPs ¶¨Á¿·ÖÎö tdTomato ÑôÐÔϸ°ûÔڷβ¿Ìض¨Ï¸°ûÀàÐÍÖеİٷֱȡ£
Ö¬ÖÊÄÉÃ׿ÅÁ£Äܹ»½«²»Í¬Ò©ÎmRNA¡¢siRNAºÍС·Ö×ÓÒ©ÎïµÈ£©°ü·â²¢¿É¿ØµØÊäË͵½ÌåÄÚµÄÌض¨²¿Î»£¬ÕâʹµÃËüÃǶÔÖÎÁƶàÖÖ¼²²¡·Ç³£ÓÐÓá£Ëæ×ÅÏà¹Ø¼¼ÊõµÄ·¢Õ¹ÏàÐÅÖ¬ÖÊÄÉÃ׿ÅÁ£»á°ïÖú¸ü¶àÒ©ÎïʵÏÖ¸ßЧµÝËÍ¡£
MCEµÄËùÓвúÆ·½öÓÃ×÷¿ÆѧÑо¿»òÒ©Ö¤É걨£¬ÎÒÃDz»ÎªÈκθöÈËÓÃ;Ìṩ²úÆ·ºÍ·þÎñ¡£
²Î¿¼ÎÄÏ×£º
[1] Hajj, K.,et al. Tools for translation: non-viral materials for therapeutic mRNA delivery. Nat Rev Mater 2, 17056 (2017).
[2] Ding BS, et al. Advanced drug delivery systems that target the vascular endothelium. Mol Interv. 2006 Apr;6(2):98-112.
[3] Hua S, et al. The use of lipid-based nanocarriers for targeted pain therapies. Front Pharmacol. 2013 Nov 21;4:143.
[4] Robson AL, et al. Advantages and Limitations of Current Imaging Techniques for Characterizing Liposome Morphology. Front Pharmacol. 2018 Feb 6;9:80.
[5] Kulkarni JA, et al. The current landscape of nucleic acid therapeutics [published correction appears in Nat Nanotechnol. 2021 Jul;16(7):841]. Nat Nanotechnol. 2021;16(6):630-643.
[6] Hou X, et al. Lipid nanoparticles for mRNA delivery. Nat Rev Mater. 2021;6(12):1078-1094.
[7] Han X, et al. An ionizable lipid toolbox for RNA delivery. Nat Commun. 2021 Dec 13;12(1):7233.
[8] Samaridou E, et al. Lipid nanoparticles for nucleic acid delivery: Current perspectives. Adv Drug Deliv Rev. 2020;154-155:37-63.
[9] Hald Albertsen C,et al. The role of lipid components in lipid nanoparticles for vaccines and gene therapy. Adv Drug Deliv Rev. 2022;188:114416.
[10] Kulkarni JA , et al . On the role of helper lipids in lipid nanoparticle formulations of siRNA. Nanoscale. 2019;11(45):21733-21739.
[11] Tenchov R, et al. Lipid Nanoparticles©¤From Liposomes to mRNA Vaccine Delivery, a Landscape of Research Diversity and Advancement. ACS Nano. 2021;15(11):16982-17015.
[12] Qiu M, et al. Lung-selective mRNA delivery of synthetic lipid nanoparticles for the treatment of pulmonary lymphangioleiomyomatosis. Proc Natl Acad Sci U S A. 2022;119(8):e2116271119. |