WO2024109612A1 - Procédé de préparation de nanoparticule lipidique pour administration efficace d'un médicament à base d'acide nucléique et utilisation associée - Google Patents
Procédé de préparation de nanoparticule lipidique pour administration efficace d'un médicament à base d'acide nucléique et utilisation associée Download PDFInfo
- Publication number
- WO2024109612A1 WO2024109612A1 PCT/CN2023/131882 CN2023131882W WO2024109612A1 WO 2024109612 A1 WO2024109612 A1 WO 2024109612A1 CN 2023131882 W CN2023131882 W CN 2023131882W WO 2024109612 A1 WO2024109612 A1 WO 2024109612A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- lipid
- nucleic acid
- ionizable
- glyceryl
- lnp
- Prior art date
Links
- 150000002632 lipids Chemical class 0.000 title claims abstract description 144
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 43
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 35
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 35
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 35
- 238000000034 method Methods 0.000 title claims abstract description 26
- 239000003814 drug Substances 0.000 title abstract description 14
- 108020004999 messenger RNA Proteins 0.000 claims abstract description 27
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 26
- 239000002131 composite material Substances 0.000 claims abstract description 19
- 150000003904 phospholipids Chemical class 0.000 claims abstract description 16
- 239000000126 substance Substances 0.000 claims abstract description 15
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 13
- 108091028075 Circular RNA Proteins 0.000 claims abstract description 3
- 108700011259 MicroRNAs Proteins 0.000 claims abstract description 3
- 108020004459 Small interfering RNA Proteins 0.000 claims abstract description 3
- 230000000692 anti-sense effect Effects 0.000 claims abstract description 3
- 239000002679 microRNA Substances 0.000 claims abstract description 3
- 239000013612 plasmid Substances 0.000 claims abstract description 3
- 238000010255 intramuscular injection Methods 0.000 claims description 25
- 239000007927 intramuscular injection Substances 0.000 claims description 25
- 239000007853 buffer solution Substances 0.000 claims description 22
- 238000002360 preparation method Methods 0.000 claims description 22
- ZISVTYVLWSZJAL-UHFFFAOYSA-N 3,6-bis[4-[bis(2-hydroxydodecyl)amino]butyl]piperazine-2,5-dione Chemical compound CCCCCCCCCCC(O)CN(CC(O)CCCCCCCCCC)CCCCC1NC(=O)C(CCCCN(CC(O)CCCCCCCCCC)CC(O)CCCCCCCCCC)NC1=O ZISVTYVLWSZJAL-UHFFFAOYSA-N 0.000 claims description 19
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 18
- 208000007514 Herpes zoster Diseases 0.000 claims description 18
- 239000008346 aqueous phase Substances 0.000 claims description 16
- 239000012074 organic phase Substances 0.000 claims description 16
- -1 [(4-hydroxybutyl)azepinediyl]bis(hexane-6,1-diyl)bis(2-hexyldecanoate) Chemical compound 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 12
- 239000002245 particle Substances 0.000 claims description 11
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims description 9
- 239000000825 pharmaceutical preparation Substances 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- OYXZMSRRJOYLLO-UHFFFAOYSA-N 7alpha-Hydroxycholesterol Natural products OC1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 OYXZMSRRJOYLLO-UHFFFAOYSA-N 0.000 claims description 4
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 claims description 4
- QCNWZROVPSVEJA-UHFFFAOYSA-N Heptadecanedioic acid Chemical compound OC(=O)CCCCCCCCCCCCCCCC(O)=O QCNWZROVPSVEJA-UHFFFAOYSA-N 0.000 claims description 4
- 238000007664 blowing Methods 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- JMOLZNNXZPAGBH-UHFFFAOYSA-M 2-hexyldecanoate Chemical compound CCCCCCCCC(C([O-])=O)CCCCCC JMOLZNNXZPAGBH-UHFFFAOYSA-M 0.000 claims description 3
- 125000000954 2-hydroxyethyl group Chemical group [H]C([*])([H])C([H])([H])O[H] 0.000 claims description 3
- SXIFAEWFOJETOA-UHFFFAOYSA-N 4-hydroxy-butyl Chemical group [CH2]CCCO SXIFAEWFOJETOA-UHFFFAOYSA-N 0.000 claims description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 3
- PSLWZOIUBRXAQW-UHFFFAOYSA-M dimethyl(dioctadecyl)azanium;bromide Chemical compound [Br-].CCCCCCCCCCCCCCCCCC[N+](C)(C)CCCCCCCCCCCCCCCCCC PSLWZOIUBRXAQW-UHFFFAOYSA-M 0.000 claims description 3
- 238000005538 encapsulation Methods 0.000 claims description 3
- 150000002148 esters Chemical class 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 229910052698 phosphorus Inorganic materials 0.000 claims description 3
- 239000011574 phosphorus Substances 0.000 claims description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 3
- QYIXCDOBOSTCEI-QCYZZNICSA-N (5alpha)-cholestan-3beta-ol Chemical compound C([C@@H]1CC2)[C@@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@H](C)CCCC(C)C)[C@@]2(C)CC1 QYIXCDOBOSTCEI-QCYZZNICSA-N 0.000 claims description 2
- PESKGJQREUXSRR-UXIWKSIVSA-N 5alpha-cholestan-3-one Chemical compound C([C@@H]1CC2)C(=O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@H](C)CCCC(C)C)[C@@]2(C)CC1 PESKGJQREUXSRR-UXIWKSIVSA-N 0.000 claims description 2
- PESKGJQREUXSRR-UHFFFAOYSA-N 5beta-cholestanone Natural products C1CC2CC(=O)CCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 PESKGJQREUXSRR-UHFFFAOYSA-N 0.000 claims description 2
- OYXZMSRRJOYLLO-RVOWOUOISA-N 7alpha-hydroxycholesterol Chemical compound C([C@H]1O)=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 OYXZMSRRJOYLLO-RVOWOUOISA-N 0.000 claims description 2
- OYXZMSRRJOYLLO-KGZHIOMZSA-N 7beta-hydroxycholesterol Chemical compound C([C@@H]1O)=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 OYXZMSRRJOYLLO-KGZHIOMZSA-N 0.000 claims description 2
- XURROWVRPGKRCH-UHFFFAOYSA-N CCCCCCCCCCOC(CCCCCN(CCCCCCCC(OC(CCCCCCCC)CCCCCCCC)=O)CCO)=O Chemical compound CCCCCCCCCCOC(CCCCCN(CCCCCCCC(OC(CCCCCCCC)CCCCCCCC)=O)CCO)=O XURROWVRPGKRCH-UHFFFAOYSA-N 0.000 claims description 2
- QYIXCDOBOSTCEI-UHFFFAOYSA-N alpha-cholestanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 QYIXCDOBOSTCEI-UHFFFAOYSA-N 0.000 claims description 2
- 229940076810 beta sitosterol Drugs 0.000 claims description 2
- LGJMUZUPVCAVPU-UHFFFAOYSA-N beta-Sitostanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 LGJMUZUPVCAVPU-UHFFFAOYSA-N 0.000 claims description 2
- NJKOMDUNNDKEAI-UHFFFAOYSA-N beta-sitosterol Natural products CCC(CCC(C)C1CCC2(C)C3CC=C4CC(O)CCC4C3CCC12C)C(C)C NJKOMDUNNDKEAI-UHFFFAOYSA-N 0.000 claims description 2
- GGCLNOIGPMGLDB-GYKMGIIDSA-N cholest-5-en-3-one Chemical compound C1C=C2CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 GGCLNOIGPMGLDB-GYKMGIIDSA-N 0.000 claims description 2
- NYOXRYYXRWJDKP-UHFFFAOYSA-N cholestenone Natural products C1CC2=CC(=O)CCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 NYOXRYYXRWJDKP-UHFFFAOYSA-N 0.000 claims description 2
- XSWSEQPWKOWORN-UHFFFAOYSA-N dodecan-2-ol Chemical compound CCCCCCCCCCC(C)O XSWSEQPWKOWORN-UHFFFAOYSA-N 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- KZJWDPNRJALLNS-VJSFXXLFSA-N sitosterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CC[C@@H](CC)C(C)C)[C@@]1(C)CC2 KZJWDPNRJALLNS-VJSFXXLFSA-N 0.000 claims description 2
- 229950005143 sitosterol Drugs 0.000 claims description 2
- KDLSKKYZXNKGTK-LQDDAWAPSA-M trimethyl-[(z)-2-[(z)-octadec-9-enoyl]-4-oxohenicos-12-enyl]azanium;chloride Chemical compound [Cl-].CCCCCCCC\C=C/CCCCCCCC(=O)CC(C[N+](C)(C)C)C(=O)CCCCCCC\C=C/CCCCCCCC KDLSKKYZXNKGTK-LQDDAWAPSA-M 0.000 claims description 2
- XWSJVWPWQMAZQM-UHFFFAOYSA-N heptadecan-9-yl octanoate Chemical compound CCCCCCCCC(CCCCCCCC)OC(=O)CCCCCCC XWSJVWPWQMAZQM-UHFFFAOYSA-N 0.000 claims 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N methyl pentane Natural products CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims 2
- 150000004702 methyl esters Chemical class 0.000 claims 1
- 230000034217 membrane fusion Effects 0.000 abstract description 6
- 230000028996 humoral immune response Effects 0.000 abstract description 3
- 125000000217 alkyl group Chemical group 0.000 abstract description 2
- 230000003213 activating effect Effects 0.000 abstract 1
- 238000007918 intramuscular administration Methods 0.000 abstract 1
- 230000021633 leukocyte mediated immunity Effects 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 42
- 102100031726 Endoplasmic reticulum junction formation protein lunapark Human genes 0.000 description 39
- 230000000052 comparative effect Effects 0.000 description 23
- 210000004027 cell Anatomy 0.000 description 21
- 241000700198 Cavia Species 0.000 description 17
- 210000002966 serum Anatomy 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 12
- 210000000952 spleen Anatomy 0.000 description 12
- 238000011740 C57BL/6 mouse Methods 0.000 description 10
- 241000699666 Mus <mouse, genus> Species 0.000 description 9
- 230000029918 bioluminescence Effects 0.000 description 9
- 238000005415 bioluminescence Methods 0.000 description 9
- 230000003053 immunization Effects 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 229940031626 subunit vaccine Drugs 0.000 description 9
- 102100037850 Interferon gamma Human genes 0.000 description 8
- 108010074328 Interferon-gamma Proteins 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 8
- 239000000427 antigen Substances 0.000 description 8
- 102000036639 antigens Human genes 0.000 description 8
- 108091007433 antigens Proteins 0.000 description 8
- 238000002649 immunization Methods 0.000 description 8
- 108700021021 mRNA Vaccine Proteins 0.000 description 8
- 229940126582 mRNA vaccine Drugs 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- 238000012795 verification Methods 0.000 description 8
- 238000001727 in vivo Methods 0.000 description 7
- 229920001223 polyethylene glycol Polymers 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 238000011725 BALB/c mouse Methods 0.000 description 6
- 238000002965 ELISA Methods 0.000 description 6
- 229940031567 attenuated vaccine Drugs 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 210000001165 lymph node Anatomy 0.000 description 6
- 229960005486 vaccine Drugs 0.000 description 6
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 210000003743 erythrocyte Anatomy 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 210000004989 spleen cell Anatomy 0.000 description 5
- 241000700199 Cavia porcellus Species 0.000 description 4
- 108090000331 Firefly luciferases Proteins 0.000 description 4
- 108010067902 Peptide Library Proteins 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- 230000007969 cellular immunity Effects 0.000 description 4
- 230000022811 deglycosylation Effects 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 4
- 230000004727 humoral immunity Effects 0.000 description 4
- 238000011503 in vivo imaging Methods 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 4
- 239000001509 sodium citrate Substances 0.000 description 4
- 210000004988 splenocyte Anatomy 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- NRJAVPSFFCBXDT-HUESYALOSA-N 1,2-distearoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCCCC NRJAVPSFFCBXDT-HUESYALOSA-N 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- DGNMJYUPWDTKJB-ZDSKVHJSSA-N bis[(z)-non-2-enyl] 9-[4-(dimethylamino)butanoyloxy]heptadecanedioate Chemical compound CCCCCC\C=C/COC(=O)CCCCCCCC(OC(=O)CCCN(C)C)CCCCCCCC(=O)OC\C=C/CCCCCC DGNMJYUPWDTKJB-ZDSKVHJSSA-N 0.000 description 3
- 230000020411 cell activation Effects 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 238000006206 glycosylation reaction Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 230000003902 lesion Effects 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 230000003248 secreting effect Effects 0.000 description 3
- HBZBAMXERPYTFS-SECBINFHSA-N (4S)-2-(6,7-dihydro-5H-pyrrolo[3,2-f][1,3]benzothiazol-2-yl)-4,5-dihydro-1,3-thiazole-4-carboxylic acid Chemical compound OC(=O)[C@H]1CSC(=N1)c1nc2cc3CCNc3cc2s1 HBZBAMXERPYTFS-SECBINFHSA-N 0.000 description 2
- 208000025721 COVID-19 Diseases 0.000 description 2
- 101000609762 Gallus gallus Ovalbumin Proteins 0.000 description 2
- 102100022297 Integrin alpha-X Human genes 0.000 description 2
- 108010002350 Interleukin-2 Proteins 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 101100233916 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) KAR5 gene Proteins 0.000 description 2
- 229920004890 Triton X-100 Polymers 0.000 description 2
- 239000013504 Triton X-100 Substances 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 230000009545 invasion Effects 0.000 description 2
- 231100000053 low toxicity Toxicity 0.000 description 2
- 230000002132 lysosomal effect Effects 0.000 description 2
- 238000012837 microfluidics method Methods 0.000 description 2
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 2
- 239000002574 poison Substances 0.000 description 2
- 231100000614 poison Toxicity 0.000 description 2
- 206010040882 skin lesion Diseases 0.000 description 2
- 231100000444 skin lesion Toxicity 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- LVNGJLRDBYCPGB-LDLOPFEMSA-N (R)-1,2-distearoylphosphatidylethanolamine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[NH3+])OC(=O)CCCCCCCCCCCCCCCCC LVNGJLRDBYCPGB-LDLOPFEMSA-N 0.000 description 1
- QFMZQPDHXULLKC-UHFFFAOYSA-N 1,2-bis(diphenylphosphino)ethane Chemical compound C=1C=CC=CC=1P(C=1C=CC=CC=1)CCP(C=1C=CC=CC=1)C1=CC=CC=C1 QFMZQPDHXULLKC-UHFFFAOYSA-N 0.000 description 1
- KILNVBDSWZSGLL-KXQOOQHDSA-N 1,2-dihexadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCC KILNVBDSWZSGLL-KXQOOQHDSA-N 0.000 description 1
- SNKAWJBJQDLSFF-NVKMUCNASA-N 1,2-dioleoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC SNKAWJBJQDLSFF-NVKMUCNASA-N 0.000 description 1
- KSXTUUUQYQYKCR-LQDDAWAPSA-M 2,3-bis[[(z)-octadec-9-enoyl]oxy]propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCC\C=C/CCCCCCCC(=O)OCC(C[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC KSXTUUUQYQYKCR-LQDDAWAPSA-M 0.000 description 1
- LRFJOIPOPUJUMI-KWXKLSQISA-N 2-[2,2-bis[(9z,12z)-octadeca-9,12-dienyl]-1,3-dioxolan-4-yl]-n,n-dimethylethanamine Chemical compound CCCCC\C=C/C\C=C/CCCCCCCCC1(CCCCCCCC\C=C/C\C=C/CCCCC)OCC(CCN(C)C)O1 LRFJOIPOPUJUMI-KWXKLSQISA-N 0.000 description 1
- UEDWDABYRNGIDC-UHFFFAOYSA-N 8-[2-hydroxyethyl-(8-nonoxy-8-oxooctyl)amino]octanoic acid Chemical compound OCCN(CCCCCCCC(=O)O)CCCCCCCC(=O)OCCCCCCCCC UEDWDABYRNGIDC-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 229940022962 COVID-19 vaccine Drugs 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 206010013700 Drug hypersensitivity Diseases 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- 241001272567 Hominoidea Species 0.000 description 1
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 1
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 1
- VKEQBMCRQDSRET-UHFFFAOYSA-N Methylone Chemical compound CNC(C)C(=O)C1=CC=C2OCOC2=C1 VKEQBMCRQDSRET-UHFFFAOYSA-N 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 241000282579 Pan Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 102000002067 Protein Subunits Human genes 0.000 description 1
- 108010001267 Protein Subunits Proteins 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 1
- OBETXYAYXDNJHR-UHFFFAOYSA-N alpha-ethylcaproic acid Natural products CCCCC(CC)C(O)=O OBETXYAYXDNJHR-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000000604 cryogenic transmission electron microscopy Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- ABCVHPIKBGRCJA-UHFFFAOYSA-N nonyl 8-[(8-heptadecan-9-yloxy-8-oxooctyl)-(2-hydroxyethyl)amino]octanoate Chemical compound OCCN(CCCCCCCC(=O)OC(CCCCCCCC)CCCCCCCC)CCCCCCCC(=O)OCCCCCCCCC ABCVHPIKBGRCJA-UHFFFAOYSA-N 0.000 description 1
- 210000004279 orbit Anatomy 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5192—Processes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
- A61K39/245—Herpetoviridae, e.g. herpes simplex virus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/5123—Organic compounds, e.g. fats, sugars
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
- A61P31/22—Antivirals for DNA viruses for herpes viruses
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/53—DNA (RNA) vaccination
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
- A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/16011—Herpesviridae
- C12N2710/16711—Varicellovirus, e.g. human herpesvirus 3, Varicella Zoster, pseudorabies
- C12N2710/16734—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Definitions
- the technical field to which the present invention belongs is the field of medical technology, and specifically relates to a lipid nanoparticle (LNP) capable of efficiently delivering nucleic acid drugs, and a preparation method and application thereof.
- LNP lipid nanoparticle
- mRNA messenger RNA
- mRNA vaccines have the advantages of high safety, short R&D cycle, high production efficiency, ability to encode multiple proteins, and no need for adjuvants.
- mRNA itself is difficult to enter cells, and mRNA itself has poor stability and is easily degraded by nucleases in the body. Therefore, a suitable delivery vehicle is needed to assist mRNA in entering target cells to work.
- Lipid nanoparticles are currently the only clinically approved mRNA vaccine carriers, which are composed of ionizable lipids, auxiliary phospholipids, cholesterol and pegylated lipids. LNP can effectively deliver mRNA into cells after intramuscular injection, activate cellular immunity and humoral immune responses, and produce effective immune protection.
- the delivery efficiency of LNP is the key to the efficacy of mRNA vaccines, and the low lysosomal escape efficiency of LNP after entering the cell is the key to limiting the delivery efficiency of LNP. Therefore, it is urgent to develop new carriers that can increase the lysosomal escape efficiency of LNP and improve the delivery efficiency of LNP.
- the present invention provides a lipid nanoparticle (hereinafter referred to as LNP), comprising a carrier and an encapsulated nucleic acid, wherein the carrier comprises a composite ionizable lipid, an auxiliary phospholipid, a cholesterol-like substance and a pegylated lipid; the nucleic acid comprises but is not limited to one or more of mRNA, circular RNA, siRNA, microRNA, antisense nucleic acid and a plasmid.
- LNP lipid nanoparticle
- the complex ionizable lipid accounts for 20 mol% to 70 mol% of the total lipids in the LNP, such as 30 mol%, 40 mol%, 50 mol%, 60 mol%.
- the complex ionizable lipid comprises a first ionizable lipid and a second ionizable lipid.
- the molar ratio of the first ionizable lipid to the second ionizable lipid is 1:99 to 99:1, preferably 2:8 to 8:2, such as 3:7, 4:6, 5:5, 6:4, 7:3, and more preferably 6:4.
- the first ionizable lipid and the second ionizable lipid are different and are independently selected from 8-[(2-hydroxyethyl)(6-oxo-6-decyloxyhexyl)amino]octanoic acid (heptadecan-9-yl) ester (SM-102), [(4-hydroxybutyl)azepinediyl]bis(hexane-6,1-diyl)bis(2-hexyldecanoate) (ALC-0315), 4(N,N-dimethylamino)butyric acid (dilinoleyl) methyl ester (DLin MC3DMA), 3,6-bis ⁇ 4-[bis(2-hydroxydodecyl)amino]butyl ⁇ piperazine-2,5-dione (cKK-E12), 9-(4-(dimethylamino)butyryloxy)heptadecanedioic
- the complex ionizable lipid includes 3,6-bis ⁇ 4-[bis(2-hydroxydodecyl)amino]butyl ⁇ piperazine-2,5-dione (cKK-E12) and 9-(4-(dimethylamino)butyryloxy)heptadecanedioic acid di((Z)-non-2-en-1-yl) ester (L319), and the molar ratio thereof is 1:99 to 99:1, preferably 2:8 to 8:2, for example 3:7, 4:6, 5:5, 6:4, 7:3.
- the auxiliary phospholipid accounts for 2 mol% to 20 mol% of the total lipids in the LNP, for example 4 mol%, 8 mol%, 10 mol%, 12 mol%, 4 mol%, 16 mol%, 18 mol%.
- the auxiliary phospholipid includes but is not limited to 1,2-distearoyl-sn-glyceryl-3-phosphatidylcholine (DSPC), 1,2-dioleoyl-sn-glyceryl-3-phosphatidylcholine (DOPC), 1,2-dipalmitoyl-sn-glyceryl-3-phosphatidylcholine (DPPC), 2-oleoyl-1-palmitoyl-sn-glyceryl-3-phosphatidylcholine (POPC), 1,2-dioleoyl-sn-glyceryl-3-phosphatidylethanolamine (DOPE), 2-oleoyl-1-palmitoyl-sn-glyceryl-3-phosphatidylethanolamine (POPE), 1,2-distearoyl-sn-glyceryl-3-phosphatidylethanolamine (DSPE), 1,2-dipalmitoyl-sn-glyceryl-3-phosphatidylcholine (DS
- the cholesterol-like substance accounts for 10 mol% to 60 mol% of the total lipids in the LNP, for example, 20 mol%, 30 mol%, 40 mol%, 50 mol%.
- the cholesterol-like substance is selected from cholesterol and its derivatives.
- the cholesterol and its derivatives include but are not limited to one or more of cholesterol, ⁇ -sitosterol, cholestanol, cholestanone, cholestenone, 7 ⁇ -hydroxycholesterol, 7 ⁇ -hydroxycholesterol, preferably cholesterol.
- the PEGylated lipid accounts for 0.3 mol% to 30 mol% of the total lipid in the LNP, preferably 0.5 mol% to 2.5 mol%, such as 1.0 mol%, 1.5 mol%, 2.0 mol%.
- the PEGylated lipids include but are not limited to one or more of 1,2-dimyristoyl-rac-glyceryl-3-methoxypolyethylene glycol (DMG-PEG), 1,2-distearoyl-rac-glyceryl-3-methoxypolyethylene glycol (DSG-PEG), 1,2-dipalmitoyl-rac-glyceryl-3-methoxypolyethylene glycol (DPG-PEG), and 1,2-distearoyl-sn-glyceryl-3-phosphatidylethanolamine-methoxypolyethylene glycol (DSPE-PEG), preferably DMG-PEG.
- DMG-PEG 1,2-dimyristoyl-rac-glyceryl-3-methoxypolyethylene glycol
- DSG-PEG 1,2-distearoyl-rac-glyceryl-3-methoxypolyethylene glycol
- DPG-PEG 1,2-dipalmitoyl-rac-glyceryl-3-me
- total lipids refers to the sum of complex ionizable lipids, auxiliary phospholipids, cholesterol substances and PEGylated lipids.
- the nucleic acid selected by the present invention includes at least one of CpG-FAM, mFLuc (mRNA encoding firefly luciferase), mOVA (mRNA encoding chicken ovalbumin), and mVZV (mRNA encoding herpes zoster gE protein).
- the encapsulation rate of the nucleic acid is 30% to 99%, preferably 70%-90%, such as 70%, 75%, 80%, 85%, 90%.
- the molar ratio of the nitrogen element contained in the complex ionizable lipid to the phosphorus element contained in the nucleic acid is (1-50):1; preferably (5-10):1, and exemplarily 5.67:1.
- the hydrated particle size of the lipid nanoparticles is less than 200 nm, such as 80 to 180 nm.
- the polydispersity coefficient of the lipid nanoparticles is less than 0.2, for example, 0.119, 0.102, or 0.093.
- the molar ratio of the complex ionizable lipid: helper phospholipid: cholesterol-like substance: polyethylene glycol lipid is 50:10:38.5:1.5.
- the composite ionizable lipid comprises a first ionizable lipid and a second ionizable lipid, the two being Examples are 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, or 2:8.
- the nucleic acid is CpG-FAM, mFLuc, mOVA or mVZV.
- the present invention also provides a method for preparing the lipid nanoparticles, which comprises preparing the lipid nanoparticles by blending an aqueous phase containing nucleic acids and a lipid organic phase through a blending method or a microfluidics method.
- the lipid organic phase is prepared as follows: the composite ionizable lipid, auxiliary phospholipid, cholesterol-like substance and pegylated lipid are dissolved in an organic solvent in proportion to obtain an organic phase.
- the organic solvent is an organic solvent that can dissolve the above substances, such as anhydrous ethanol.
- the preparation method of the aqueous phase containing nucleic acid is as follows: dissolving the nucleic acid in a buffer solution to obtain an aqueous phase.
- the buffer solution can be a buffer solution with a pH of 4 to 6, for example, a sodium citrate buffer solution with a pH of 4 or 5.5.
- the blending method includes the following steps: using a pipette to absorb the aqueous phase containing nucleic acids, quickly adding it to the lipid organic phase, and then quickly blowing the mixed liquid for more than 30 times, blowing it evenly and letting it stand at room temperature for 10 minutes, and dialyzing the resulting product after standing, for example, dialyzing it in a PBS buffer solution with a volume greater than 1000 times for more than 6 hours to obtain the lipid nanoparticles.
- the microfluidic method includes the following steps: fixing syringes containing an aqueous phase and a lipid organic phase on a syringe pump respectively, injecting the aqueous phase: lipid organic phase into a microfluidic chip at a volume ratio of 3:1 and mixing thoroughly, collecting the mixed solution after mixing and standing at room temperature for 10 minutes, and dialyzing the resulting product with a PBS buffer solution after standing, for example, dialyzing in a PBS buffer solution with a volume greater than 1000 times for more than 6 hours to obtain the lipid nanoparticles.
- the present invention also provides the use of the lipid nanoparticles in the preparation of pharmaceutical preparations or biological preparations.
- the biologic is an injectable biologic.
- the biologic is a vaccine, preferably an mRNA vaccine.
- the administration method of the biological preparation is intramuscular injection.
- the biological agent is used to prevent herpes zoster.
- the present invention also provides a pharmaceutical preparation or a biological preparation having the meaning as described above.
- the present invention also provides a method for preventing and/or treating a disease, comprising administering a preventively and/or therapeutically effective amount of the above-mentioned pharmaceutical preparation or biological product to a subject; for example, the disease is herpes zoster.
- the term "effective amount" can be determined according to the method mastered by a physician with clinical practice qualifications in the field to determine the amount of the pharmaceutical preparation or biological product of the present invention that is sufficient to achieve the intended application (including but not limited to the treatment of diseases defined above). Determining the effective dose for prevention and/or treatment is within the capabilities of clinicians or researchers in the field and may be changed due to the following factors: the intended application (in vitro or in vivo), or the subject and disease condition to be treated, such as the subject's weight and age, general health, severity of the disease condition, mode of administration, and other factors that affect the efficacy, such as a history of drug allergies.
- the specific dosage will vary depending on the following factors: the specific biological product selected, the dosage regimen based on, whether it is co-administered with other drugs, the timing of administration, the tissue to which it is administered, and the physical delivery system carried.
- the "subject" of the present invention refers to a specific human or other warm-blooded mammal.
- the human as “subject” of the present invention includes adults, infants, and children, and other warm-blooded mammals include but are not limited to non-human primates, such as chimpanzees, other apes or monkeys, and other zoo animals, domestic mammals or laboratory animals, such as cats, pigs, dogs, cows, sheep, mice, rats, and guinea pigs.
- the LNP prepared by the present invention has uniform particle size, good dispersibility, high encapsulation efficiency, good biocompatibility, low toxicity and side effects, and can be highly
- the LNP preparation method of the present invention has simple operation steps and is easy to mass produce.
- the present invention combines a first ionizable lipid and a second ionizable lipid to form a composite ionizable lipid, which has an ionizable lipid with high efficiency in entering cells and an ionizable lipid with a long alkyl chain and high efficiency in membrane fusion ability.
- the composite ionizable lipid of the present invention can significantly improve the mRNA expression efficiency of the commercial four-component LNP through synergistic effect.
- the present invention uses composite ionizable lipids to prepare five-component LNPs, which have a higher activation efficiency on antigen-presenting cells in draining lymph nodes and spleen than commercial four-component LNPs after intramuscular injection.
- the five-component LNP prepared by the composite ionizable lipids of the present invention can efficiently deliver the herpes zoster mRNA vaccine. After intramuscular injection, it significantly activates the cellular immunity and humoral immune response in the body, produces a large number of antigen-specific binding antibodies in the blood, and increases the antigen-specific T cell content in the spleen, which has an excellent protective effect. It is expected to achieve a breakthrough in the field of herpes zoster mRNA vaccines and has great application prospects.
- FIG. 1 Chemical structures of the first ionizable lipid and the second ionizable lipid used in the examples of the present invention.
- FIG. 2 Hydration particle size distribution curve of LNP obtained in Example 1B of the present invention.
- FIG3 Cryo-TEM image of the five-component LNP obtained in Example 1A of the present invention.
- FIG. 4 shows the bioluminescence intensity of DC2.4 cells transfected with LNPs loaded with mFluc in Example 3 of the present invention.
- FIG5 The membrane fusion ability of the five-component LNP was detected using mouse red blood cells in Example 4 of the present invention.
- Example 5 of the present invention in vivo imaging of small animals was used to verify the bioluminescence intensity of the five-component LNP loaded with mFLuc at the injection site of mice; (a) In vivo imaging image and statistical results of bioluminescence intensity at the injection site 4 hours after intramuscular injection; (b) In vivo imaging image and statistical results of bioluminescence intensity at the injection site 24 hours after intramuscular injection.
- Figure 7 Flow chart of animal experiment in Example 6 of the present invention.
- FIG8 Activation efficiency of different LNPs loaded with mOVA on DC cells in draining lymph nodes and spleen in Example 6 of the present invention.
- Figure 9 Flow chart of animal experiment in Example 7 of the present invention.
- Figure 10A Antigen-specific Total IgG antibody titers in the serum of mice on days 14, 28, and 35 after the first intramuscular injection of LNPs loaded with mVZV in Example 7 of the present invention.
- Figure 11 The number of antigen-specific splenocytes secreting IFN- ⁇ in the spleen of mice on the 42nd day after the first intramuscular injection of LNPs loaded with mVZV in Example 7 of the present invention (a) ELISpot spot number statistics; (b) optical photograph of the ELISpot well.
- Figure 12 The number of antigen-specific CD4 + T cells secreting IFN- ⁇ in the spleen of mice on day 42 after the first intramuscular injection of LNPs loaded with mVZV in Example 7 of the present invention.
- Figure 13 Concentrations of IL-2 and IFN- ⁇ secreted by antigen-specific spleen cells on day 42 after the first intramuscular injection of LNPs encapsulating mVZV in mice in Example 7 of the present invention.
- Figure 14 Body weight monitoring curve of mice during immunization in Example 8 of the present invention.
- FIG. 15 H&E staining of tissue and organ sections on day 42 after the first intramuscular injection of LNPs loaded with mVZV in mice in Example 8 of the present invention.
- Figure 16 Flow chart of animal experiment in Example 9 of the present invention.
- Figure 17 Total serum levels at 2, 4 and 6 weeks after the first intramuscular injection of LNPs loaded with mVZV in mice in Example 9 of the present invention Antibody titers of IgG, IgG1, IgG2a, and IgG2c.
- Figure 18 Total IgG antibody titer in the serum of mice during 8 months after the first intramuscular injection of LNPs loaded with mVZV in Example 9 of the present invention.
- Figure 19 The number of specific splenocytes secreting IFN- ⁇ in the spleen of mice on the 42nd day after the first intramuscular injection of LNPs loaded with mVZV in Example 9 of the present invention (a) optical photograph of ELISpot wells; (b) statistical graph of the number of ELISpot spots.
- Figure 20 Body weight monitoring curve of mice during immunization process in Example 9 of the present invention.
- Figure 21 Schematic diagram of the deglycosylation site in Example 10 of the present invention.
- Figure 22 Total IgG, IgG1, IgG2a and IgG2c antibody titers in the serum of mice at 2, 4 and 6 weeks after the first intramuscular injection of LNPs loaded with mVZV in Example 10 of the present invention.
- Figure 23 Flow chart of animal experiments in Example 11 of the present invention.
- Figure 24 A diagram of the skin changes on the back of a guinea pig after infection with a poison in Example 11 of the present invention.
- Figure 25 Guinea pig skin lesion scoring table in Example 11 of the present invention.
- Figure 26 Skin lesion scores on the back of guinea pigs after poison challenge in Example 11 of the present invention.
- Figure 27 H&E staining of the skin tissue of the guinea pig's back 7 days after infection in Example 11 of the present invention.
- Figure 28 Skin viral load on the back of guinea pigs 7 days after infection in Example 11 of the present invention.
- Figure 29 Body weight monitoring curve of guinea pigs during immunization in Example 11 of the present invention.
- MC in the above figures represents the product of Comparative Example 1, i.e., a four-component LNP with MC3 as the ionizable lipid;
- cKK-E12 represents the product of Comparative Example 2, i.e., a four-component LNP with cKK-E12 as the ionizable lipid;
- L319 represents the product of Comparative Example 3, i.e., a four-component LNP with L319 as the ionizable lipid.
- the composite ionizable lipid includes a first ionizable lipid 3,6-bis ⁇ 4-[bis(2-hydroxydodecyl)amino]butyl ⁇ piperazine-2,5-dione (abbreviated as cKK-E12), and a second ionizable lipid 9-(4-(dimethylamino)butyryloxy)heptadecanedioic acid di((Z)-non-2-en-1-yl) ester (abbreviated as L319);
- the auxiliary phospholipid is 1,2-distearoyl-sn-glyceryl-3-phosphocholine, abbreviated as DSPC;
- the polyethylene glycol lipid is 1,2-dimyristoyl-sn-glyceryl-3-methoxypolyethylene glycol 2000, abbreviated as DMG-PEG2000.
- the structural formulas of the first ionizable lipid and the second ionizable lipid
- Five-component lipid nanoparticles (referred to as five-component LNPs) were prepared as follows:
- the preparation method of the five-component lipid nanoparticles of this embodiment is basically the same as that of Example 1A, wherein the nucleic acid drug is mRNA encoding firefly luciferase (mFLuc), and the molar ratio of cKK-E12 and L319 is 2:8, 3:7, 4:6, 5:5, 6:4, 7:3 or 8:2.
- the nucleic acid drug is mRNA encoding firefly luciferase (mFLuc)
- molar ratio of cKK-E12 and L319 is 2:8, 3:7, 4:6, 5:5, 6:4, 7:3 or 8:2.
- the preparation method of the five-component lipid nanoparticles of this example is basically the same as that of Example 1A, except that the nucleic acid drug is mRNA encoding chicken ovalbumin (mOVA).
- the nucleic acid drug is mRNA encoding chicken ovalbumin (mOVA).
- the preparation method of the five-component lipid nanoparticles of this embodiment is basically the same as that of Example 1A, except that the nucleic acid drug is mRNA encoding the herpes zoster virus gE protein (mVZV).
- the nucleic acid drug is mRNA encoding the herpes zoster virus gE protein (mVZV).
- Comparative Example 1 the composite ionizable lipid was replaced with MC3 (i.e., 4(N,N-dimethylamino)butyric acid (dilinoleyl) methyl ester), i.e., a four-component LNP with MC3 as the ionizable lipid;
- MC3 i.e., 4(N,N-dimethylamino)butyric acid (dilinoleyl) methyl ester
- Comparative Example 2 the composite ionizable lipid was replaced with cKK-E12 (i.e., the molar ratio of cKK-E12 to L319 was 1:0), i.e., a four-component LNP with cKK-E12 as the ionizable lipid;
- the hydrated particle size and polydispersity index of the five-component LNP of Example 1A are 140 nm and 0.09, respectively, the hydrated particle size and polydispersity index of the five-component LNP of Example 1C are 110 nm and 0.1, respectively, and the hydrated particle size and polydispersity index of the five-component LNP of Example 1D are 130 nm and 0.1, respectively.
- the hydrated particle sizes of the five-component lipid nanoparticles prepared in Examples 1A to 1D are all less than 200 nm, and the polydispersity coefficient is less than 0.2, indicating that the preparation method of the present invention can form stable lipid nanoparticles with uniform particle size.
- the five-component LNPs loaded with mFLuc prepared in Example 1B were used, and the four-component LNPs of Comparative Examples 1 to 3 were selected as a control group to screen different ratios of cKK-E12 and L319 as ionizable lipids to act synergistically.
- the experimental steps are as follows: DC2.4 cells were incubated overnight in a 96-well plate at 20,000 cells per well. When the cell confluence reached 50%, LNPs of Example 1B and Comparative Examples 1 to 3 were added respectively, and cultured in a saturated humidity incubator at 37°C and 5% CO2 concentration for 24 hours. An Untreated cells group (referring to cells without any treatment after plating) and a Free mRNA group (referring to mRNA without a carrier being added directly to the cell culture medium) were also set up. The culture medium was removed, cell lysate and firefly luciferase substrate were added, and after shaking for 10 minutes, the bioluminescence value was read using an ELISA reader. The test results are shown in Figure 4.
- Example 1A The mFLuc-encapsulated five-component LNP prepared in Example 1A was used, and the four-component LNP of Comparative Examples 1 to 3 was selected as a control group to detect the membrane fusion ability of the five-component LNP.
- mice erythrocytes are placed in a transparent 96-well plate, sodium citrate buffer solution of pH 5.5 and PBS buffer solution of pH 7.4 are added respectively, and then LNPs of Example 1 and Comparative Examples 1 to 3 are added correspondingly and incubated at 37°C for 1 hour.
- a negative control group i.e., only buffer solution and erythrocytes are added to the well plate, recorded as Untreated cells
- a positive control group i.e., buffer solution, erythrocytes and 0.5% Triton X-100 are added to the well plate, recorded as 0.5% Triton X-100 are incubated at 37°C for 1 hour.
- Example 1A The mFLuc-encapsulated five-component LNP prepared in Example 1A was used, and the four-component LNP of Comparative Examples 1 to 3 were selected as a control group to detect the mRNA expression efficiency of the five-component LNP of the present invention in vivo.
- mice Female C57BL/6 mice aged 6-8 weeks were selected, and 2 ⁇ g of LNP of Example 1A and Comparative Examples 1 to 3 were administered to each mouse in each group by intramuscular injection. 200 ⁇ L of 200 mg/mL firefly luciferase substrate was injected intraperitoneally into each mouse 4 hours and 24 hours after administration, respectively. After 15 minutes, the bioluminescence intensity in the mice was detected using small animal in vivo imaging.
- the five-component LNP of the present invention has a high mRNA expression efficiency in vivo.
- Example 1C The mOVA-encapsulated five-component LNP prepared in Example 1C was used, wherein the four-component LNP of Comparative Example 1 was selected as a control group, and the DC cell activation efficiency of the five-component LNP of the present invention in C57BL/6 mice was detected.
- mice 8-week-old female C57BL/6 mice were selected, with 4 mice in each experimental group. Each group of mice was immunized by intramuscular injection on day 0 and day 7, and each mouse in each group was administered 10 ⁇ g of LNP of Example 1C and Comparative Example 1, respectively. A PBS group was also set up in which only PBS buffer solution was injected; on day 8, the mice were killed, and the draining lymph nodes and spleen at the injection site were removed in vitro. For the experimental process, see Figure 7.
- lymph node cells and spleen cells were processed separately, a single cell suspension was obtained, and the number of activated DC cells (CD11c + CD80 + , CD11c + CD86 + ) in the spleen and draining lymph nodes was detected by flow cytometry, see Figure 8.
- the mRNA-loaded five-component LNP of the present invention has a high DC cell activation efficiency in C57BL/6 mice in the draining lymph nodes and spleen of mice.
- Example 7 Verification of the immune efficacy of five-component LNP in C57BL/6 mice
- the five-component LNPs encapsulating mVZV prepared in Example 1D were used, wherein the four-component LNPs of Comparative Example 1, the attenuated VZV virus vaccine (hereinafter referred to as the attenuated vaccine) and the herpes zoster recombinant gE protein subunit vaccine plus AS01 adjuvant (hereinafter referred to as the subunit vaccine) were selected as the control groups of this experiment to detect the immune efficacy of the five-component LNPs of the present invention in C57BL/6 mice.
- the attenuated vaccine the attenuated VZV virus vaccine
- AS01 adjuvant hereinafter referred to as the subunit vaccine
- mice 8-week-old female C57BL/6 mice were selected, with 4 mice in each experimental group. Each group of mice was immunized by intramuscular injection at week 0 and week 3. Each mouse in each group was administered 10 ⁇ g of LNP, attenuated vaccine and subunit vaccine of Example 1D and Comparative Example 1, respectively. A PBS group was also set up to be injected with only PBS buffer solution. Blood was collected from the mice at week 2, 4 and 5, and serum was separated. The mice were killed at week 6. The experimental process is shown in Figure 9. The ELISA was used to measure the 14th, 28th and 35th day of the mice. The test results of the antibody titers of herpes zoster-specific IgG, IgG1, and IgG2c in serum are shown in FIG. 10A to FIG. 10C .
- the number of herpes zoster-specific T cells (CD3 + CD4 + CD45 + IFN- ⁇ + ) was detected by flow cytometry, and the test results are shown in Figure 12.
- Splenocytes were then placed in a 96-well plate with 1 million cells per well, stimulated with the herpes zoster gE peptide library for 48 hours, centrifuged to obtain the cell supernatant, and the amount of IL-2 and IFN- ⁇ secreted by spleen cells in the cell supernatant was measured by ELISA, and the test results are shown in Figure 13.
- the mRNA-loaded five-component LNP of the present invention has the ability to efficiently activate specific cellular immunity and humoral immunity in C57BL/6 mice.
- Example 8 Verification of the toxicity of five-component LNP in vivo
- the five-component LNPs loaded with mVZV prepared in Example 1D were used, and the four-component LNPs, attenuated vaccine and subunit vaccine of Comparative Example 1 were selected as the control groups of this experiment to detect the vaccine efficacy of the five-component LNPs in vivo.
- mice 8-week-old female C57BL/6 mice were selected, with 4 mice in each experimental group. Each group of mice was immunized by intramuscular injection at week 0 and week 3. Each mouse in each group was administered 10 ⁇ g of the five-component LNP in Example 1D and the four-component LNP, attenuated vaccine and subunit vaccine in Comparative Example 1, respectively.
- a PBS group was set up in which only PBS buffer solution was injected. The weight of the mice was monitored during the immunization period. See Figure 14 for the experimental process. The mice were killed in week 6, and the heart, liver, spleen, lung, kidney, brain and injection site muscles of the mice were embedded in paraffin for H&E staining sections. See Figure 15 for the test results.
- the five-component LNPs loaded with mVZV prepared in Example 1D were used, wherein the four-component LNPs of Comparative Example 1, the attenuated vaccine and the subunit vaccine were selected as the control groups of this experiment to detect the vaccine efficacy of the five-component LNPs in BALB/c mice.
- mice 8-week-old female BALB/c mice were selected, with 3 mice in each experimental group, and the mice were immunized by intramuscular injection at week 0 and week 3, respectively.
- Each mouse was administered 10 ⁇ g of the five-component LNP in Example 1D and the four-component LNP, attenuated vaccine and subunit vaccine in Comparative Example 1, respectively.
- Orbital blood was drawn from the mice at weeks 2, 4 and 6, the serum was separated, and the mice were killed at week 6. See Figure 16 for the experimental process.
- ELISA was used to measure the antibody titers of Total IgG, IgG1, IgG2a and IgG2c specific to herpes zoster in the serum of mice at 2, 4 and 6 weeks, see Figure 17.
- ELISA was used to measure the changes in the titers of Total IgG specific to herpes zoster in the serum of mice within 8 months after immunization, see Figure 18.
- the spleen of the mice was processed to obtain a single cell suspension, and 300,000 cells were placed in each well of the ELISpot plate and stimulated with the herpes zoster gE peptide library for 24 hours, and then the number of spleen cells that secreted IFN- ⁇ specifically for herpes zoster was detected by color development, see Figure 19 (a) ELISpot well optical photograph, (b) ELISpot spot number statistical chart. The weight of the mice was monitored during the immunization period, see Figure 20.
- Example 10 Verification of the effect of five-component LNPs loaded with mVZV of different deglycosylation designs on humoral immunity
- mVZV in Example 1D was used to perform different deglycosylation site designs, including: two nitrogen-glycosylation site mutations (abbreviated as mMUTE-N), six oxygen-glycosylation site mutations (abbreviated as mMUTE-O), and a combination of two glycosylation site mutations (abbreviated as mMUTE-N+O), see Figure 21.
- mMUTE-N nitrogen-glycosylation site mutations
- mMUTE-O oxygen-glycosylation site mutations
- mMUTE-N+O a combination of two glycosylation site mutations
- Example 9 Referring to the immune experiment process and experimental steps in Example 9, specifically including: selecting 8-week-old female BALB/c mice, 6 mice in each experimental group, immunizing the mice by intramuscular injection at week 0 and week 3, and administering 10 ⁇ g of the five-component LNP in Example 1D and the five-component LNP containing mMUTE-N, the five-component LNP containing mMUTE-O, and the five-component LNP containing mMUTE-N+O prepared above, respectively, which are recorded as VZV, mMUTE-N, mMUTE-O, and mMUTE-N+O, respectively, and collecting blood from the eye sockets of the mice at weeks 2, 4, and 6 to separate the serum.
- Example 11 Verification of the five-component LNP in guinea pigs against VZV virus invasion
- the five-component LNPs containing mVZV prepared in Example 1D were used, and subunit vaccines and PBS were selected as the control group of this experiment to detect the efficacy of the five-component LNPs in guinea pigs against the virus.
- the experimental process is shown in Figure 23. The experimental steps are specifically as follows: guinea pigs weighing 200-250g were selected, and the guinea pigs were immunized by intramuscular injection on days 0 and 14. The guinea pigs were administered 10 ⁇ g of the five-component LNPs, subunit vaccines and PBS in Example 1D, respectively. The guinea pigs were scratched on the back on days 21 and 22 for the challenge experiment.
- Example 1D The body weight of guinea pigs was monitored during the immunization period, see Figure 29. It was verified that the five-component LNP encapsulated with mVZV in Example 1D can effectively protect guinea pigs from VZV virus invasion.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Nanotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Virology (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Crystallography & Structural Chemistry (AREA)
- Biomedical Technology (AREA)
- Optics & Photonics (AREA)
- Biophysics (AREA)
- Communicable Diseases (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Oncology (AREA)
- Medical Informatics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- General Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Manufacturing & Machinery (AREA)
- General Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- Dermatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Preparation (AREA)
Abstract
L'invention concerne une nanoparticule lipidique pour administrer efficacement un médicament à base d'acide nucléique, un procédé de préparation et une utilisation associée. La nanoparticule lipidique comprend un vecteur et un acide nucléique encapsulé. Le vecteur comprend un lipide ionisable composite, un phospholipide auxiliaire, une substance de cholestérol et un lipide pégylé. L'acide nucléique comprend un ou plusieurs éléments parmi l'ARNm, l'ARN circulaire, l'ARNsi, le miARN, un acide nucléique antisens et un plasmide. Le lipide ionisable composite comprend un lipide ionisable ayant la capacité d'entrer efficacement dans des cellules, et un lipide ionisable avec une longue chaîne alkyle et la capacité de fusion membranaire efficace. Le lipide ionisable composite peut améliorer significativement l'efficacité d'expression d'ARNm de LNPs à quatre composants commerciaux au moyen d'un synergisme. La nanoparticule lipidique facilite l'expression efficace du médicament à base d'acide nucléique piégé dans celle-ci après administration intramusculaire, activant ainsi efficacement des réponses immunitaires humorales et à médiation cellulaire.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211477861.X | 2022-11-23 | ||
CN202211477861.XA CN118059061A (zh) | 2022-11-23 | 2022-11-23 | 一种用于高效递送核酸药物的脂质纳米颗粒的制备方法和应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2024109612A1 true WO2024109612A1 (fr) | 2024-05-30 |
Family
ID=91096100
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2023/131882 WO2024109612A1 (fr) | 2022-11-23 | 2023-11-15 | Procédé de préparation de nanoparticule lipidique pour administration efficace d'un médicament à base d'acide nucléique et utilisation associée |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN118059061A (fr) |
WO (1) | WO2024109612A1 (fr) |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111467321A (zh) * | 2020-03-26 | 2020-07-31 | 深圳市新合生物医疗科技有限公司 | 一种mRNA核酸类药物胞内递送系统、制备方法和应用 |
CN114099699A (zh) * | 2021-11-22 | 2022-03-01 | 华南理工大学 | 一种纳米递送系统及其制备方法和应用 |
CN114306279A (zh) * | 2021-12-30 | 2022-04-12 | 复旦大学 | 基于科罗索酸或其类似物的脂质纳米颗粒系统及其制备方法和应用 |
WO2022207862A2 (fr) * | 2021-03-31 | 2022-10-06 | Curevac Ag | Seringues contenant des compositions pharmaceutiques comprenant de l'arn |
CN116763756A (zh) * | 2022-10-11 | 2023-09-19 | 中国科学院化学研究所 | 一种脂质纳米颗粒及其制备方法和应用 |
-
2022
- 2022-11-23 CN CN202211477861.XA patent/CN118059061A/zh active Pending
-
2023
- 2023-11-15 WO PCT/CN2023/131882 patent/WO2024109612A1/fr unknown
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111467321A (zh) * | 2020-03-26 | 2020-07-31 | 深圳市新合生物医疗科技有限公司 | 一种mRNA核酸类药物胞内递送系统、制备方法和应用 |
WO2022207862A2 (fr) * | 2021-03-31 | 2022-10-06 | Curevac Ag | Seringues contenant des compositions pharmaceutiques comprenant de l'arn |
CN114099699A (zh) * | 2021-11-22 | 2022-03-01 | 华南理工大学 | 一种纳米递送系统及其制备方法和应用 |
CN114306279A (zh) * | 2021-12-30 | 2022-04-12 | 复旦大学 | 基于科罗索酸或其类似物的脂质纳米颗粒系统及其制备方法和应用 |
CN116763756A (zh) * | 2022-10-11 | 2023-09-19 | 中国科学院化学研究所 | 一种脂质纳米颗粒及其制备方法和应用 |
Also Published As
Publication number | Publication date |
---|---|
CN118059061A (zh) | 2024-05-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP4800485B2 (ja) | ポリヌクレオチドベースのワクチンに対する免疫応答を増強するためのアジュバント組成物および方法 | |
US20200016274A1 (en) | Messenger rna vaccines and uses thereof | |
CN110974954A (zh) | 一种用于增强核酸疫苗免疫效果的脂质纳米颗粒及其制备方法 | |
CN107980004A (zh) | 用于医治疾病的外泌体的用途 | |
US20230226169A1 (en) | Multilamellar rna nanoparticle vaccine against sars-cov-2 | |
WO2024078193A1 (fr) | Nanoparticule lipidique, son procédé de préparation et son utilisation | |
CN116133640A (zh) | 脂质纳米颗粒 | |
CN115103687A (zh) | Hpv疫苗 | |
WO2023147092A9 (fr) | Vaccin anti-coronavirus | |
CN115252582A (zh) | 红细胞膜杂合pH脂质体包被溶瘤病毒制剂的制备及应用 | |
EP3866769A1 (fr) | Nanotransporteurs pour une thérapie d'inflammation pulmonaire | |
Zhou et al. | Single-injection subunit vaccine for rabies prevention using lentinan as adjuvant | |
Austin et al. | Split-dose administration enhances immune responses elicited by a mRNA/lipid nanoparticle vaccine expressing respiratory syncytial virus F protein | |
WO2024109612A1 (fr) | Procédé de préparation de nanoparticule lipidique pour administration efficace d'un médicament à base d'acide nucléique et utilisation associée | |
JP2023181377A (ja) | 免疫賦活剤 | |
US20040005302A1 (en) | Encapsulated cells to elicit immune responses | |
Oh et al. | Preclinical safety assessment of the suction-assisted intradermal injection of the SARS-CoV-2 DNA vaccine candidate pGO-1002 in white rabbit | |
Colombani et al. | Harnessing the Potential of Biomaterials for COVID-19 Therapeutic Strategies | |
CN118146318B (zh) | 一种腺相关病毒衣壳蛋白及其用途 | |
US20050232906A1 (en) | Adjuvant preparation for the induction of specific immunity | |
US20230302112A1 (en) | Respiratory synctial virus rna vaccine | |
WO2024108955A1 (fr) | Vaccin contre l'hépatite b | |
WO2023169500A1 (fr) | Vaccin à arnm pour le codage de la protéine s du sars-cov-2 | |
WO2023183945A1 (fr) | Compositions et procédés pour vaccins adjuvantés | |
CN118574637A (zh) | Rna分子 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 23893702 Country of ref document: EP Kind code of ref document: A1 |