CN118388389A - Endogenous amino acid derived ionizable lipid, and preparation method and application thereof - Google Patents
Endogenous amino acid derived ionizable lipid, and preparation method and application thereof Download PDFInfo
- Publication number
- CN118388389A CN118388389A CN202410477849.1A CN202410477849A CN118388389A CN 118388389 A CN118388389 A CN 118388389A CN 202410477849 A CN202410477849 A CN 202410477849A CN 118388389 A CN118388389 A CN 118388389A
- Authority
- CN
- China
- Prior art keywords
- lipid
- amino acid
- formula
- endogenous amino
- derived
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 150000002632 lipids Chemical class 0.000 title claims abstract description 96
- 150000001413 amino acids Chemical class 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 239000003814 drug Substances 0.000 claims abstract description 43
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 27
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 26
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 26
- -1 lipid compound Chemical class 0.000 claims abstract description 23
- 238000012377 drug delivery Methods 0.000 claims abstract description 8
- 230000002829 reductive effect Effects 0.000 claims abstract description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 5
- 229920001184 polypeptide Polymers 0.000 claims abstract description 4
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 54
- 239000002105 nanoparticle Substances 0.000 claims description 39
- 150000001875 compounds Chemical class 0.000 claims description 36
- 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 description 34
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 33
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- 239000000203 mixture Substances 0.000 claims description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 18
- 238000006243 chemical reaction Methods 0.000 claims description 18
- 235000012000 cholesterol Nutrition 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 18
- 229940079593 drug Drugs 0.000 claims description 16
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 16
- 108020004999 messenger RNA Proteins 0.000 claims description 13
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 12
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 12
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 11
- 108020004414 DNA Proteins 0.000 claims description 9
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 9
- 239000004698 Polyethylene Substances 0.000 claims description 9
- MWRBNPKJOOWZPW-CLFAGFIQSA-N dioleoyl phosphatidylethanolamine Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(COP(O)(=O)OCCN)OC(=O)CCCCCCC\C=C/CCCCCCCC MWRBNPKJOOWZPW-CLFAGFIQSA-N 0.000 claims description 9
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 8
- 239000003054 catalyst Substances 0.000 claims description 8
- 239000012074 organic phase Substances 0.000 claims description 8
- 239000003960 organic solvent Substances 0.000 claims description 8
- 229920000573 polyethylene Polymers 0.000 claims description 8
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 8
- 108090000623 proteins and genes Proteins 0.000 claims description 8
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 7
- 239000003480 eluent Substances 0.000 claims description 7
- 239000000126 substance Substances 0.000 claims description 7
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 6
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- 150000001204 N-oxides Chemical class 0.000 claims description 6
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 6
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 6
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 6
- 230000009471 action Effects 0.000 claims description 6
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 claims description 6
- YGPSJZOEDVAXAB-UHFFFAOYSA-N kynurenine Chemical compound OC(=O)C(N)CC(=O)C1=CC=CC=C1N YGPSJZOEDVAXAB-UHFFFAOYSA-N 0.000 claims description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 230000035484 reaction time Effects 0.000 claims description 6
- 150000003839 salts Chemical class 0.000 claims description 6
- 239000012453 solvate Substances 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- IRUJRVAXEALDLJ-UHFFFAOYSA-N tert-butyl 4-[2-(methylamino)ethyl]piperazine-1-carboxylate Chemical compound CNCCN1CCN(C(=O)OC(C)(C)C)CC1 IRUJRVAXEALDLJ-UHFFFAOYSA-N 0.000 claims description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N ethylene glycol Natural products OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 5
- AOCSUUGBCMTKJH-UHFFFAOYSA-N tert-butyl n-(2-aminoethyl)carbamate Chemical compound CC(C)(C)OC(=O)NCCN AOCSUUGBCMTKJH-UHFFFAOYSA-N 0.000 claims description 5
- 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 claims description 4
- SLKDGVPOSSLUAI-PGUFJCEWSA-N 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine zwitterion Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OCCN)OC(=O)CCCCCCCCCCCCCCC SLKDGVPOSSLUAI-PGUFJCEWSA-N 0.000 claims description 4
- 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 claims 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 claims description 4
- LVNGJLRDBYCPGB-UHFFFAOYSA-N 1,2-distearoylphosphatidylethanolamine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(COP([O-])(=O)OCC[NH3+])OC(=O)CCCCCCCCCCCCCCCCC LVNGJLRDBYCPGB-UHFFFAOYSA-N 0.000 claims description 4
- BIABMEZBCHDPBV-MPQUPPDSSA-N 1,2-palmitoyl-sn-glycero-3-phospho-(1'-sn-glycerol) Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@@H](O)CO)OC(=O)CCCCCCCCCCCCCCC BIABMEZBCHDPBV-MPQUPPDSSA-N 0.000 claims description 4
- BDNKZNFMNDZQMI-UHFFFAOYSA-N 1,3-diisopropylcarbodiimide Chemical compound CC(C)N=C=NC(C)C BDNKZNFMNDZQMI-UHFFFAOYSA-N 0.000 claims description 4
- JQKOHRZNEOQNJE-ZZEZOPTASA-N 2-azaniumylethyl [3-octadecanoyloxy-2-[(z)-octadec-9-enoyl]oxypropyl] phosphate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(COP([O-])(=O)OCC[NH3+])OC(=O)CCCCCCC\C=C/CCCCCCCC JQKOHRZNEOQNJE-ZZEZOPTASA-N 0.000 claims description 4
- 108091028075 Circular RNA Proteins 0.000 claims description 4
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 claims description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 4
- OFBQJSOFQDEBGM-UHFFFAOYSA-N Pentane Chemical compound CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 claims description 4
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 4
- 108020004459 Small interfering RNA Proteins 0.000 claims description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 claims description 4
- 108091046869 Telomeric non-coding RNA Proteins 0.000 claims description 4
- DSNRWDQKZIEDDB-GCMPNPAFSA-N [(2r)-3-[2,3-dihydroxypropoxy(hydroxy)phosphoryl]oxy-2-[(z)-octadec-9-enoyl]oxypropyl] (z)-octadec-9-enoate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCC\C=C/CCCCCCCC DSNRWDQKZIEDDB-GCMPNPAFSA-N 0.000 claims description 4
- MVPPADPHJFYWMZ-UHFFFAOYSA-N chlorobenzene Chemical compound ClC1=CC=CC=C1 MVPPADPHJFYWMZ-UHFFFAOYSA-N 0.000 claims description 4
- JHIVVAPYMSGYDF-UHFFFAOYSA-N cyclohexanone Chemical compound O=C1CCCCC1 JHIVVAPYMSGYDF-UHFFFAOYSA-N 0.000 claims description 4
- BGRWYRAHAFMIBJ-UHFFFAOYSA-N diisopropylcarbodiimide Natural products CC(C)NC(=O)NC(C)C BGRWYRAHAFMIBJ-UHFFFAOYSA-N 0.000 claims description 4
- STVZJERGLQHEKB-UHFFFAOYSA-N ethylene glycol dimethacrylate Chemical compound CC(=C)C(=O)OCCOC(=O)C(C)=C STVZJERGLQHEKB-UHFFFAOYSA-N 0.000 claims description 4
- 230000002068 genetic effect Effects 0.000 claims description 4
- 108091070501 miRNA Proteins 0.000 claims description 4
- 239000002679 microRNA Substances 0.000 claims description 4
- 239000013612 plasmid Substances 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 102000004169 proteins and genes Human genes 0.000 claims description 4
- 238000010898 silica gel chromatography Methods 0.000 claims description 4
- 239000004055 small Interfering RNA Substances 0.000 claims description 4
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims description 3
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 3
- 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 claims description 3
- 150000003384 small molecules Chemical class 0.000 claims description 3
- OCJBOOLMMGQPQU-UHFFFAOYSA-N 1,4-dichlorobenzene Chemical compound ClC1=CC=C(Cl)C=C1 OCJBOOLMMGQPQU-UHFFFAOYSA-N 0.000 claims description 2
- FHQVHHIBKUMWTI-ZCXUNETKSA-N 1-palmitoyl-2-oleoyl phosphatidylethanolamine Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCCN)OC(=O)CCCCCCC\C=C/CCCCCCCC FHQVHHIBKUMWTI-ZCXUNETKSA-N 0.000 claims description 2
- SYBYTAAJFKOIEJ-UHFFFAOYSA-N 3-Methylbutan-2-one Chemical compound CC(C)C(C)=O SYBYTAAJFKOIEJ-UHFFFAOYSA-N 0.000 claims description 2
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 claims description 2
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 claims description 2
- 239000007821 HATU Substances 0.000 claims description 2
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 claims description 2
- UIHCLUNTQKBZGK-UHFFFAOYSA-N Methyl isobutyl ketone Natural products CCC(C)C(C)=O UIHCLUNTQKBZGK-UHFFFAOYSA-N 0.000 claims description 2
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 2
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 2
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 2
- GOOHAUXETOMSMM-UHFFFAOYSA-N Propylene oxide Chemical compound CC1CO1 GOOHAUXETOMSMM-UHFFFAOYSA-N 0.000 claims description 2
- 239000012317 TBTU Substances 0.000 claims description 2
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims description 2
- CLZISMQKJZCZDN-UHFFFAOYSA-N [benzotriazol-1-yloxy(dimethylamino)methylidene]-dimethylazanium Chemical compound C1=CC=C2N(OC(N(C)C)=[N+](C)C)N=NC2=C1 CLZISMQKJZCZDN-UHFFFAOYSA-N 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 125000000217 alkyl group Chemical group 0.000 claims description 2
- 238000007112 amidation reaction Methods 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- WZWSOGGTVQXXSN-UHFFFAOYSA-N cyclohexanone;toluene Chemical compound CC1=CC=CC=C1.O=C1CCCCC1 WZWSOGGTVQXXSN-UHFFFAOYSA-N 0.000 claims description 2
- 229940117389 dichlorobenzene Drugs 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 238000005886 esterification reaction Methods 0.000 claims description 2
- 238000001704 evaporation Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 claims description 2
- 150000004676 glycans Chemical class 0.000 claims description 2
- 229940116336 glycol dimethacrylate Drugs 0.000 claims description 2
- 229910052739 hydrogen Inorganic materials 0.000 claims description 2
- 239000001257 hydrogen Substances 0.000 claims description 2
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 2
- TVMXDCGIABBOFY-UHFFFAOYSA-N octane Chemical compound CCCCCCCC TVMXDCGIABBOFY-UHFFFAOYSA-N 0.000 claims description 2
- 229910052760 oxygen Inorganic materials 0.000 claims description 2
- 150000008104 phosphatidylethanolamines Chemical class 0.000 claims description 2
- 239000005017 polysaccharide Substances 0.000 claims description 2
- 229920001282 polysaccharide Polymers 0.000 claims description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 2
- 229910052717 sulfur Inorganic materials 0.000 claims description 2
- 239000008096 xylene Substances 0.000 claims description 2
- 239000012295 chemical reaction liquid Substances 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000001415 gene therapy Methods 0.000 abstract description 5
- 238000001727 in vivo Methods 0.000 abstract description 4
- 230000001988 toxicity Effects 0.000 abstract description 3
- 231100000419 toxicity Toxicity 0.000 abstract description 3
- 108090000790 Enzymes Proteins 0.000 abstract description 2
- 102000004190 Enzymes Human genes 0.000 abstract description 2
- 239000008346 aqueous phase Substances 0.000 abstract description 2
- 238000006731 degradation reaction Methods 0.000 abstract description 2
- 230000002708 enhancing effect Effects 0.000 abstract description 2
- 230000002209 hydrophobic effect Effects 0.000 abstract description 2
- 230000002434 immunopotentiative effect Effects 0.000 abstract description 2
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 239000002502 liposome Substances 0.000 abstract description 2
- 230000001766 physiological effect Effects 0.000 abstract description 2
- 150000003862 amino acid derivatives Chemical class 0.000 abstract 1
- 230000015556 catabolic process Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 23
- 101150068408 lnp1 gene Proteins 0.000 description 9
- 238000001890 transfection Methods 0.000 description 9
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 6
- 239000011541 reaction mixture Substances 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 5
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 5
- 238000004440 column chromatography Methods 0.000 description 5
- 239000000741 silica gel Substances 0.000 description 5
- 229910002027 silica gel Inorganic materials 0.000 description 5
- MYMSJFSOOQERIO-UHFFFAOYSA-N 1-bromodecane Chemical compound CCCCCCCCCCBr MYMSJFSOOQERIO-UHFFFAOYSA-N 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 229940126214 compound 3 Drugs 0.000 description 4
- 229940125898 compound 5 Drugs 0.000 description 4
- 238000005538 encapsulation Methods 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 230000008685 targeting Effects 0.000 description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 229940125904 compound 1 Drugs 0.000 description 3
- 229940125782 compound 2 Drugs 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000010992 reflux Methods 0.000 description 3
- 238000004809 thin layer chromatography Methods 0.000 description 3
- 239000003981 vehicle Substances 0.000 description 3
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- 102000007330 LDL Lipoproteins Human genes 0.000 description 2
- 108010007622 LDL Lipoproteins Proteins 0.000 description 2
- 108700001237 Nucleic Acid-Based Vaccines Proteins 0.000 description 2
- 102000004338 Transferrin Human genes 0.000 description 2
- 108090000901 Transferrin Proteins 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- 230000036571 hydration Effects 0.000 description 2
- 238000006703 hydration reaction Methods 0.000 description 2
- 230000002163 immunogen Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000001000 micrograph Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 229940023146 nucleic acid vaccine Drugs 0.000 description 2
- 239000012044 organic layer Substances 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000002390 rotary evaporation Methods 0.000 description 2
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000010409 thin film Substances 0.000 description 2
- 239000012581 transferrin Substances 0.000 description 2
- 229960004799 tryptophan Drugs 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- PBLNBZIONSLZBU-HNHCFKFXSA-N 1-bromododecane Chemical group CCCCCCCCCCC[13CH2]Br PBLNBZIONSLZBU-HNHCFKFXSA-N 0.000 description 1
- VMKOFRJSULQZRM-UHFFFAOYSA-N 1-bromooctane Chemical compound CCCCCCCCBr VMKOFRJSULQZRM-UHFFFAOYSA-N 0.000 description 1
- KOFZTCSTGIWCQG-UHFFFAOYSA-N 1-bromotetradecane Chemical group CCCCCCCCCCCCCCBr KOFZTCSTGIWCQG-UHFFFAOYSA-N 0.000 description 1
- 108091023037 Aptamer Proteins 0.000 description 1
- 108010002913 Asialoglycoproteins Proteins 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- NFVNYBJCJGKVQK-ZDUSSCGKSA-N N-[(Tert-butoxy)carbonyl]-L-tryptophan Chemical compound C1=CC=C2C(C[C@H](NC(=O)OC(C)(C)C)C(O)=O)=CNC2=C1 NFVNYBJCJGKVQK-ZDUSSCGKSA-N 0.000 description 1
- LFZAGIJXANFPFN-UHFFFAOYSA-N N-[3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-thiophen-2-ylpropyl]acetamide Chemical compound C(C)(C)C1=NN=C(N1C1CCN(CC1)CCC(C=1SC=CC=1)NC(C)=O)C LFZAGIJXANFPFN-UHFFFAOYSA-N 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical group 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 239000010408 film Substances 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 238000001476 gene delivery Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000002132 lysosomal effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- QTYWBJZOZDYCGB-UHFFFAOYSA-L potassium;sodium;2-carboxybenzoate;hydroxide Chemical compound [OH-].[Na+].[K+].OC(=O)C1=CC=CC=C1C([O-])=O QTYWBJZOZDYCGB-UHFFFAOYSA-L 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 125000005629 sialic acid group Chemical group 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the technical field of biological medicine, and particularly relates to endogenous amino acid derived ionizable lipid, and a preparation method and application thereof. The invention provides an endogenous amino acid derivative ionizable lipid compound shown in a formula (I), which is based on endogenous amino acid, so that toxicity is greatly reduced, and safety is remarkably improved. The nucleic acid drug delivery device comprises a hydrophilic amino acid head part and a hydrophobic fatty chain tail part, has obvious amphipathy, can form a liposome structure in an aqueous phase, and can effectively deliver the nucleic acid drug into cells. The structure of the polypeptide contains an amide bond, and the bond can be rapidly hydrolyzed by enzymes in vivo, is easy to metabolize and remove, and has biodegradability. Amino acids produced during degradation play a role in immunopotentiating or enhancing/inhibiting other physiological activities. This property makes this novel lipid compound have great potential in the field of drug delivery and is expected to become an important carrier for future gene therapy and drug delivery.Formula (I).
Description
Technical Field
The invention belongs to the technical field of biological medicine, and particularly relates to endogenous amino acid derived ionizable lipid, and a preparation method and application thereof.
Background
The disclosure of this background section is only intended to increase the understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art.
Gene therapy (GENE THERAPY) is a therapeutic method for correcting or compensating diseases caused by defective or abnormal genes by introducing exogenous genes into target cells. Nucleic acid vaccine (nucleic ACID VACCINE), also called genetic vaccine (GENETICVACCINE), refers to a nucleic acid sequence (such as DNA, mRNA, etc.) containing encoded immunogenic protein or polypeptide is introduced into a host, expressed by the host cell, and the host cell is induced to generate immune response to the immunogen, so as to achieve the purpose of preventing and treating diseases. Among them, ensuring the smooth introduction of foreign genes is an extremely important part of gene therapy and immunization with genetic vaccines. Among the many methods of gene delivery, methods of developing suitable lipid nanoparticles (i.e., LNP, lipid Nanoparticle) to encapsulate nucleic acids, target them to target cells, and deliver nucleic acids of specific genes into cells are increasingly being used by scientists.
One obvious difference between nucleic acid drugs and common chemical drugs is that nucleic acids carry a large number of phosphates, thus being negatively charged and of large molecular weight. In order to enable better encapsulation by LNP, various lipid compounds such as ionizable lipids have been developed.
Ionizable lipids typically contain one or more ionizable amine groups in the molecular structure, the apparent pKa of which is a key attribute for nucleic acid drug delivery in vivo. The chemical space of the ionizable lipid structure and the great difference of the ionizable lipid structure in cell uptake and endosomal escape exist, so that the development of the ionizable lipid with different chemical spaces, biodegradability and high transfection efficiency has great significance for promoting the deep development and clinical transformation of nucleic acid medicaments.
Disclosure of Invention
In order to overcome the problems, the invention provides an endogenous amino acid derived ionizable lipid, a preparation method and application thereof.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
In a first aspect of the present invention there is provided an endogenous amino acid derived ionizable lipid or an N-oxide, solvate, pharmaceutically acceptable salt or stereoisomer thereof, having the structural formula (i):
Wherein R 1 and R 2 are the same or different from each other and are each independently selected from substituted or unsubstituted linear or branched C 8~C20 alkyl, or substituted or unsubstituted linear or branched C 8~C20 alkenyl, or substituted or unsubstituted linear or branched C 8~C20 alkynyl, wherein R 3、R4 can be H;
R 3 and R 4 are the same or different from each other and are each independently selected from hydrogen, substituted or unsubstituted linear or branched C 8~C20 alkyl, or substituted or unsubstituted linear or branched C 8~C20 alkenyl, or substituted or unsubstituted linear or branched C 8~C20 alkynyl;
x is O, NH or S;
m is a positive integer of 1 to 3;
L 1 is A group.
In one or more embodiments, L 1 isWhen the amino acid is a tryptophan, the endogenous amino acid is tryptophan.
In one or more embodiments, L 1 isWhen the group is, the endogenous amino acid is kynurenine.
In one or more embodiments, the endogenous amino acid-derived ionizable lipid is selected from one of the following compounds:
in a second aspect of the present invention, there is provided a method for preparing an endogenous amino acid-derived ionizable lipid as described above, comprising:
In an organic solvent, under the action of a catalyst, a compound shown in a formula (II) and a compound shown in a formula (III) are subjected to esterification reaction or amidation reaction to obtain a compound shown in a formula (I);
In the compounds of the formula (II) and the compounds of the formula (III), R 1、R2、R3、R4、X、L1 and m have the same meanings as in the compounds of the formula (I).
In one or more embodiments, when X is nitrogen, m=1, and r 1、R2 is independently selected from C 10~14 alkyl, a method of preparing a compound represented by formula (iii) includes the steps of: in acetonitrile, under the action of potassium carbonate, N-tert-butoxycarbonyl-1, 2-ethylenediamine and bromoalkane react to obtain an intermediate a; reacting a dichloromethane solution of the intermediate a under the action of trifluoroacetic acid to obtain an intermediate b, namely a compound shown in a formula (III);
the specific process is as follows:
Preferably, the volume ratio of the mole amount of the N-t-butoxycarbonyl-1, 2-ethylenediamine to the acetonitrile is 0.01-1 mole/L; the molar ratio of the potassium carbonate to the N-tert-butoxycarbonyl-1, 2-ethylenediamine is 1-3:1; the mol ratio of the N-tert-butyloxycarbonyl-1, 2-ethylenediamine to the bromoalkane is 1:1-1.5; the reaction temperature of the N-tert-butoxycarbonyl-1, 2-ethylenediamine and bromoalkane is 60-100 ℃ and the reaction time is 60-80 h; the molar ratio of the intermediate a to the trifluoroacetic acid is 1-3:1; the reaction temperature of the intermediate a for producing the intermediate b is ice bath, and the reaction time is 2-8 h.
In one or more embodiments, the organic solvent is selected from one or more of methanol, ethanol, isopropanol, benzene, toluene, xylene, pentane, hexane, octane, cyclohexane, cyclohexanone, toluene cyclohexanone, chlorobenzene, dichlorobenzene, methylene chloride, diethyl ether, propylene oxide, acetone, methyl butanone, methyl isobutyl ketone, acetonitrile, pyridine, phenol, styrene, perchloroethylene, trichloroethylene, dimethyl sulfoxide, ethylene glycol ether, N-dimethylformamide, or triethanolamine;
preferably, when the solvent is methanol, dichloromethane, acetonitrile, ethyl acetate, dimethyl sulfoxide; the volume ratio of the compound shown in the formula (II) to the organic solvent is 0.01-10 mol/L.
In one or more embodiments, the catalyst is selected from one or more of N-hydroxysuccinimide (NHS), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI), 1-Hydroxybenzotriazole (HOBT), 2- (7-azobenzotriazole) -N, N '-tetramethylurea Hexafluorophosphate (HATU), O-benzotriazol-tetramethylurea Hexafluorophosphate (HBTU), 4-Dimethylaminopyridine (DMAP), N-Diisopropylethylamine (DIPEA), or O-benzotriazol-N-, N' -tetramethylurea tetrafluoroboric acid (TBTU);
preferably, when the catalyst is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI), 1-Hydroxybenzotriazole (HOBT) and N, N-Diisopropylethylamine (DIPEA), the molar ratio of the catalyst to the compound represented by formula (II) is 1-1.5:1.
In one or more embodiments, the molar ratio of the compound of formula (II) to the compound of formula (III) is from 1:1 to 1.5; preferably 1:1.2.
In one or more embodiments, the reaction temperature is room temperature and the reaction time is 10 to 30 hours.
In one or more embodiments, the method for post-treating a reaction solution obtained by reacting a compound represented by formula (II) with a compound represented by formula (III) comprises the steps of:
adding saturated sodium chloride solution and saturated sodium bicarbonate solution into the reaction solution, extracting with dichloromethane, collecting an organic phase, drying the organic phase by anhydrous sodium sulfate, filtering, evaporating under reduced pressure, and separating by silica gel column chromatography to obtain lipid; the eluent used for the silica gel column chromatography is a mixture of dichloromethane and methanol, DCM/meoh=200:0 to 10:1.
In a third aspect of the invention there is provided the use of an endogenous amino acid derived ionizable lipid of the first aspect or an N-oxide, solvate, pharmaceutically acceptable salt or stereoisomer thereof as a drug delivery vehicle.
In a fourth aspect of the invention there is provided a lipid nanoparticle composition comprising an endogenous amino acid-derived ionizable lipid of the first aspect, a helper lipid, cholesterol, a PEG lipid, and a entrapped drug.
In one or more embodiments, the auxiliary lipid is selected from a variety of sources including distearoyl phosphatidylcholine (DSPC), dioleoyl phosphatidylcholine (DOPC), dipalmitoyl phosphatidylcholine (DPPC), dioleoyl phosphatidylglycerol (DOPG), dipalmitoyl phosphatidylglycerol (DPPG), dioleoyl phosphatidylethanolamine (DOPE), palmitoyl phosphatidylcholine (POPC), 1-palmitoyl-2-oleoyl phosphatidylethanolamine (POPE), dioleoyl phosphatidylethanolamine-maleimide (DOPE-MAL), dipalmitoyl phosphatidylethanolamine (DPPE), dipyristoyl phosphatidylcholine (DMPE), distearoyl phosphatidylethanolamine (DSPE), and 1-stearoyl-2-oleoyl phosphatidylethanolamine (SOPE), preferably dioleoyl phosphatidylethanolamine (DOPE).
In one or more embodiments, the cholesterol is selected from one or both of cholesterol or 10α -hydroxycholesterol; preferably, the cholesterol is cholesterol or a mixture of cholesterol and 10 alpha-hydroxycholesterol (mass ratio 1:1).
In one or more embodiments, the PEG lipid is selected from polyethylene glycol-dimyristoyl glycerol (PEG-DMG), polyethylene glycol-distearoyl phosphatidylethanolamine (PEG-DSPE), polyethylene glycol-dimethacrylate (PEG-DMA); polyethylene glycol-dimyristoylglycerol (PEG-DMG) is preferred.
In one or more embodiments, the encapsulated drug comprises one or more of a biological drug or a chemical drug; the biological medicine comprises one or more of nucleic acid medicine, protein medicine, polypeptide medicine or polysaccharide medicine; preferably, the nucleic acid drug comprises one or more of messenger RNA (mRNA), small interfering RNA (siRNA), micro RNA (microRNA), circular RNA (circRNA), long non-coding RNA (lncRNA), plasmid DNA (plasmid DNA) and small circle DNA (mini circle DNA); the chemical medicine comprises one or more of small molecule medicine, fluorescein or developer; further preferably, the nucleic acid agent is mRNA.
In one or more embodiments, the lipid nanoparticle composition has a diameter of 1nm to 1000nm; preferably 100 to 200nm.
In one or more embodiments, the endogenous amino acid derived ionizable lipid, helper lipid, cholesterol, PEG lipid of the first aspect is in a molar ratio of 15-45:20-35:10-40:0.5-5;
The mass ratio of the endogenous amino acid derived ionizable lipid to the encapsulated drug of the first aspect is 1-100:1.
In one or more embodiments, the targeting molecule can be modified on the lipid nanoparticle composition to provide targeting functions to target a particular cell, tissue or organ. The targeting molecule may be in the whole lipid nanoparticle composition or may be located only on the surface of the lipid nanoparticle composition. The targeting molecule may be a protein, peptide, glycoprotein, lipid, small molecule, nucleic acid, etc., examples of which include, but are not limited to, antibodies, antibody fragments, low Density Lipoproteins (LDL), transferrin (transferrin), asialoglycoprotein (asialycoprotein), receptor ligands, sialic acids, aptamers, etc.
In one or more embodiments, the lipid nanoparticle composition can be prepared using a variety of methods known in the art, including, but not limited to, thin film hydration, extrusion, nano-precipitation, and microfluidic methods, among others. Preferred methods of preparation include, but are not limited to, the following: film hydration method: the lipid is dissolved in an organic solvent to form a thin film, which is then hydrated by the addition of water or a buffer solution to form nanoparticles. Nano precipitation method: the lipid and nucleic acid drug are dissolved in an organic solvent and an aqueous phase, respectively, and then the two phases are mixed to form nanoparticles by precipitation. Microfluidic method: the lipid and the nucleic acid solution are mixed in a microfluidic manner by utilizing a microfluidic technology, and the nano-particles are prepared through the control of a microchannel. Further preferred, the lipid nanoparticle composition is prepared by microfluidic methods.
In a fifth aspect of the present invention, there is provided the use of the lipid nanoparticle composition of the fourth aspect for the preparation of a genetic medicament comprising an active ingredient and a delivery vehicle, the active ingredient being a nucleic acid medicament, the delivery vehicle being the lipid nanoparticle composition of the fourth aspect.
The invention has the beneficial effects that:
(1) The invention provides an endogenous amino acid derived ionizable lipid compound, which is based on endogenous amino acid, greatly reduces toxicity and remarkably improves safety. The compound comprises a hydrophilic amino acid head part and a hydrophobic fatty chain tail part, has obvious amphipathy, can form a liposome structure in a water phase, and can effectively deliver nucleic acid medicines into cells. In addition, the secondary compound has an amide bond in the structure, and the bond can be rapidly hydrolyzed by enzymes in vivo, is easy to metabolize and remove, and has biodegradability. Amino acids produced by this degradation process play a role in immunopotentiating or enhancing/inhibiting other physiological activities. This property makes this novel lipid compound have great potential in the field of drug delivery and is expected to become an important carrier for future gene therapy and drug delivery.
(2) The preparation method of the endogenous amino acid ionizable lipid compound provided by the invention is based on the simple reaction of common carboxylic acid functional groups and organic amine compounds, and has the advantages of easily obtained raw materials, scientific synthetic route, simple operation steps, mild reaction conditions and high product separation efficiency.
(3) In the lipid nanoparticle composition, the structure and proportion of ionizable lipids have a significant impact on the lysosomal escape capacity of the nucleic acid; the ratio of helper lipid to ionizable lipid can also have an effect; the choice of different phospholipids and the proportion of ionizable lipids also affect the transfection efficiency of nucleic acids. In order to achieve efficient transfection of nucleic acids, the invention defines that the molar ratio of endogenous amino acid ionizable lipid, auxiliary lipid, cholesterol and PEG lipid is 15-45:20-35:10-40:0.5-5; the mass ratio of the endogenous amino acid ionizable lipid to the drug is 1-100:1.
(4) The endogenous amino acid derived ionizable lipid provided by the invention can effectively deliver nucleic acid drugs, can realize efficient transfection of nucleic acids in vivo and in vitro, and has the advantages of greatly reducing toxicity and remarkably improving safety based on endogenous amino acids, so that the endogenous amino acid derived ionizable lipid has wide application prospects in delivery of nucleic acid drugs.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention.
FIG. 1 a shows the synthetic routes of the compounds 1 to 3 in examples 1 to 3 according to the invention, b shows the synthetic route in example 4, c shows the synthetic route of the compound 5;
FIG. 2 is a graph showing particle diameters (a) and Zeta potentials (b) of lipid nanoparticles LNP-1 to 5;
FIG. 3 shows the encapsulation efficiency of lipid nanoparticle LNP-3;
FIG. 4 is a transmission electron microscope image of lipid nanoparticle LNP-3
FIG. 5 shows the results of evaluation of transfection efficiency of lipid nanoparticles in the present invention.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present invention. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.
In order to enable those skilled in the art to more clearly understand the technical scheme of the present invention, the technical scheme of the present invention will be described in detail with reference to specific embodiments.
In FIG. 1, a is the synthetic route for the compounds 1 to 3 in examples 1 to 3, b is the synthetic route in example 4, and c is the synthesis of the compound 5 in example 5.
EXAMPLE 1 Synthesis of Compound 1
(1) Synthesis of intermediate N1, N1-didecyl-1, 2-ethylenediamine (intermediate b (Z11 b)):
Into a 250mL round bottom flask equipped with a magneton, t-butoxycarbonyl-1, 2-ethylenediamine (10 mmol), 1-bromodecane (22 mmol), potassium carbonate (20 mmol) and acetonitrile (60 mL) were added and heated under reflux at 80℃for 72h. After the reaction is finished, the reaction solution is filtered by suction, the solid is removed, the solvent is removed by rotary evaporation under reduced pressure, and the product is separated and purified by column chromatography (200-300 meshes of silica gel, eluent: methanol/dichloromethane volume ratio=1:20), thus obtaining intermediate a (Z11 a), and the yield is 75%.
Trifluoroacetic acid (2.5 mL) was added to a solution of intermediate a (Z11 a) (300.65 mg,7 mmol) in glacial dichloromethane (6 mL) and stirred for 6 hours. To the reaction mixture was added saturated sodium bicarbonate solution (5 mL). The organic layer was separated, washed with saturated sodium bicarbonate (3×10 mL) and saturated sodium chloride solution (3×10 mL), and the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporator to give intermediate b (Z11 b) in 97% yield.
(2) Intermediate b (Z11 b) (691 mg,2.27 mmol) was weighed and dissolved in 20mL of dichloromethane. N-Boc-L-tryptophan (268 mg,2.72 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI) (803 mg,2.73 mmol), 1-Hydroxybenzotriazole (HOBT) (268 mg,2.72 mmol), N-Diisopropylethylamine (DIPEA) (351 mg,2.72 mmol) were added to the solution and stirred at room temperature for 17h. The reaction was monitored by thin layer chromatography. The reaction mixture was washed with saturated brine (3×10 mL), and the organic phase was dried over anhydrous sodium sulfate. After concentration on a rotary evaporator, purification by column chromatography (column: 200-300 mesh silica gel; eluent: DCM/meoh=100:0-10:1) afforded intermediate c (936 mg, 65.73% yield).
Trifluoroacetic acid (2.5 mL) was added to a solution of intermediate c (936 mg,1.49 mmol) in ice dichloromethane (6 mL) and stirred for 6 hours. To the reaction mixture was added saturated sodium bicarbonate solution (5 mL). The organic layer was separated, washed with saturated sodium bicarbonate (3×10 mL) and saturated sodium chloride solution (3×10 mL), and the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporator to give compound 1 in 95% yield.
1H NMR(400MHz,Chloroform-d)δ8.15(s,1H),7.61(dd,J=61.4,6.2Hz,2H),7.38-7.06(m,4H),3.70(dd,J=9.0,4.2Hz,1H),3.49-3.23(m,3H),2.90(dd,J=14.4,9.0Hz,1H),2.41(dt,J=42.7,6.8Hz,6H),1.25(d,J=3.5Hz,32H),0.88(t,J=6.7Hz,6H).
EXAMPLE 2 Synthesis of Compound 2
(1) N1, N1, -Didodecyl ethane-1, 2-diamine (intermediate b (Z12 b)) and intermediate a (Z12 a) and were synthesized as described in example 1, except that 1-bromododecane was replaced with 1-bromodecane (22 mol); other steps and conditions were consistent with example 1. The single step yield of intermediate a (Z12 a) was 78%.
(2) The intermediate b (Z11 b) in example 1 was replaced with Z12b in this example, and the other conditions were exactly the same as in example 1, to synthesize compound 2, and finally yield 896mg of compound 2 was obtained in 61%.
1H NMR(400MHz,Chloroform-d)δ8.15(s,1H),7.61(dd,J=61.4,6.2Hz,2H),7.38-7.06(m,4H),3.70(dd,J=9.0,4.2Hz,1H),3.49-3.23(m,3H),2.90(dd,J=14.4,9.0Hz,1H),2.41(dt,J=42.7,6.8Hz,6H),1.25(d,J=3.5Hz,42H),0.88(t,J=6.7Hz,6H).
Example 3
N1, N1, -Bitetradecylethane-1, 2-diamine (intermediate b (Z13 b)) and intermediate a (Z13 a) were synthesized as described in example 1, except that 1-bromotetradecane was replaced by 1-bromodecane (22 mol); other steps and conditions were consistent with example 1. The single step yield of intermediate a (Z13 a) was 79%.
(2) The intermediate b (Z11 b) in example 1 was replaced with Z13b in this example, and the other conditions were exactly the same as those in example 1, to thereby synthesize compound 3, and finally yield 931mg of compound 3 was obtained in 59%.
1H NMR(400MHz,Chloroform-d)δ8.15(s,1H),7.61(dd,J=61.4,6.2Hz,2H),7.38-7.06(m,4H),3.70(dd,J=9.0,4.2Hz,1H),3.49-3.23(m,3H),2.90(dd,J=14.4,9.0Hz,1H),2.41(dt,J=42.7,6.8Hz,6H),1.25(d,J=3.5Hz,50H),0.88(t,J=6.7Hz,6H).
Example 4
Synthesis of intermediate N, N, -didodecyl-ethanolamine (intermediate d)
To an acetonitrile solution (18 mL) of ethanolamine (3.6 mmol) were added 1-bromodecane (8 mmol), potassium carbonate (16 mmol) and potassium iodide (0.4 mmol). The reaction mixture was stirred at 80℃under reflux for 48 hours. The reaction was monitored by thin layer chromatography. The reaction mixture was cooled to room temperature, filtered to remove potassium carbonate and potassium iodide, and concentrated under reduced pressure. After concentrating to dryness, purification by column chromatography (200-300 mesh silica gel, eluent: dichloromethane/methanol=50:1-20:1) afforded N, -didodecyl-ethanolamine, intermediate d (1.14 g, 92% yield).
Intermediate d (410 mg,1.2 mmol) was weighed and dissolved in 5mL DMSO. Kynurenine (213 mg,1 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI) (230 mg,1.2 mmol), 1-Hydroxybenzotriazole (HOBT) (162 mg,1.2 mmol) 9, triethylamine (607 mg,6 mmol) were added to the solution and stirred at room temperature for 17h. The reaction was monitored by thin layer chromatography. The reaction mixture was washed with saturated brine (3×10 mL), and the organic phase was dried over anhydrous sodium sulfate. After concentration by rotary evaporator, the mixture was purified by column chromatography (column: 200-300 mesh silica gel; eluent: PE/ea=100:0-3:1) to give compound 4 (339 mg, yield 64%).
1H NMR(400MHz,Chloroform-d)δ7.52-7.61(m,1H),7.45(dd,J=7.9,1.4Hz,1H),7.22(td,J=7.6,1.2Hz,1H),6.86(dd,J=8.2,1.1Hz,1H),4.35(t,J=6.9Hz,2H),3.99(t,J=7.5Hz,1H),2.87(t,J=6.9Hz,2H),2.7-2.8(m,4H),2.73(s,2H),1.37(p,J=2.7Hz,4H),1.22(ddt,J=11.0,8.9,6.4Hz,16H),-3.77(s,18H),0.78-0.86(m,7H).
Example 5
Compound 3 (10 mmol), 1-bromooctane (22 mmol), potassium carbonate (20 mmol) and acetonitrile (60 ml) were weighed and heated under reflux at 80℃for 48h. After the reaction, the reaction solution is filtered by suction, the solid is removed, the solvent is removed by rotary evaporation under reduced pressure, and the product is separated and purified by column chromatography (200-300 meshes of silica gel, eluent: methanol/dichloromethane volume ratio=1:20), thus obtaining the compound 5 with the yield of 75%.
1H NMR(400MHz,Chloroform-d)δ8.15(s,1H),7.61(dd,J=61.4,6.2Hz,2H),7.38-7.06(m,3H),3.70(dd,J=9.0,4.2Hz,1H),3.49-3.23(m,3H),3.15(dd,J=10.1,9.0Hz,1H),,2.90(dd,J=14.4,9.0Hz,1H),2.45(m,6.8Hz,10H),1.27(d,J=3.6Hz,72H),0.88(m,12H).
Experimental example 1
Lipid nanoparticle in vitro delivery mRNA performance test
Preparation of lipid nanoparticles: the prepared amino acid ionizable lipid (compounds 1-5), cholesterol, DMG-PEG and DOPE are dissolved in ethanol according to the molar ratio of 15:25:1:20 to prepare a lipid ethanol solution (wherein the molar concentration of the amino acid ionizable lipid is 10 mg/mL). mRNA of Green Fluorescent Protein (GFP) was dissolved in potassium hydrogen phthalate-sodium hydroxide buffer at pH=4 to give mRNA solution (10 ng/. Mu.L). Lipid ethanol solution and mRNA solution were prepared into amino acid ionizable lipids using NanoAssemblr microfluidic device (Precision NanoSystems company): rapidly mixing the mRNA in a weight ratio of 10:1 to prepare a solution containing lipid nanoparticles; after ethanol was removed by dialysis of the solution containing lipid nanoparticles, lipid nanoparticles LNP-1 to 5 (LNP-1 (Compound 1), LNP-2 (Compound 2), LNP-3 (Compound 3), LNP-4 (Compound 4), and LNP-5 (Compound 5), respectively) were obtained.
Characterization of the lipid nanoparticles LNP-1 to 5 obtained was performed, and FIG. 2 is a graph showing particle diameters (a) and Zeta potentials (b) of the lipid nanoparticles LNP-1 to 5. FIG. 3 shows the encapsulation efficiency of the lipid nanoparticle LNP-3, and FIG. 4 shows a transmission electron microscope image of the lipid nanoparticle LNP-3.
As can be seen from FIG. 2, the particle size of the lipid nanoparticles LNP-1 to 5 is within 200nm, and the potential is slightly negatively charged. From FIG. 3, it can be seen that the encapsulation efficiency of the lipid nanoparticle LNP-3 is good (90.+ -. 1.06%). From FIG. 4 it can be seen that the lipid nanoparticle LNP-3 is spheroidized.
Evaluation of transfection efficiency of lipid nanoparticles LNP-1 to 5 in Hep3B cell line:
Hep3B cells in logarithmic growth phase were inoculated into 24-well plates containing DMEM medium at a density of 1×10 4/well, cultured overnight at 37 ℃, fresh medium containing lipid nanoparticle LNP-1-5 (containing 300ng mRNA) was added to each well, the Control group was added with the same volume of PBS, and after incubation for 12 hours at 37 ℃, the transfection efficiency was measured by flow cytometry and was high, and the result is shown in FIG. 5.
As can be seen from fig. 5, although the difference in particle diameters of the lipid nanoparticles LNP-1 to 5 is small, there is a large difference in delivery efficiency. The highest transfection efficiency of LNP-3 is 89+ -2.1%, and the transfection efficiencies of LNP-1, LNP-2, LNP-4, LNP-5 are 83.9+ -1.6%, 79.6+ -1.35%, 59.1+ -3.1%, 71.7+ -2.3%, respectively.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. An endogenous amino acid-derived ionizable lipid or an N-oxide, solvate, pharmaceutically acceptable salt or stereoisomer thereof, characterized in that the endogenous amino acid-derived ionizable lipid has a structural formula represented by formula (i):
Wherein R 1 and R 2 are the same or different from each other and are each independently selected from substituted or unsubstituted linear or branched C 8~C20 alkyl, or substituted or unsubstituted linear or branched C 8~C20 alkenyl, or substituted or unsubstituted linear or branched C 8~C20 alkynyl, wherein R 3、R4 can be H;
R 3 and R 4 are the same or different from each other and are each independently selected from hydrogen, substituted or unsubstituted linear or branched C 8~C20 alkyl, or substituted or unsubstituted linear or branched C 8~C20 alkenyl, or substituted or unsubstituted linear or branched C 8~C20 alkynyl;
x is O, NH or S;
m is a positive integer of 1 to 3;
L 1 is A group.
2. An endogenous amino acid-derived ionizable lipid or an N-oxide, solvate, pharmaceutically acceptable salt or stereoisomer thereof according to claim 1, wherein when L 1 isWhen the amino acid is a group, the endogenous amino acid is tryptophan;
when L 1 is When the group is, the endogenous amino acid is kynurenine.
3. The endogenous amino acid-derived ionizable lipid or an N-oxide, solvate, pharmaceutically acceptable salt or stereoisomer thereof according to claim 1, wherein the endogenous amino acid-derived ionizable lipid is selected from one of the following compounds:
4. a method of preparing an endogenous amino acid-derived ionizable lipid according to claim 1, comprising:
In an organic solvent, under the action of a catalyst, a compound shown in a formula (II) and a compound shown in a formula (III) are subjected to esterification reaction or amidation reaction to obtain a compound shown in a formula (I);
In the compounds of the formula (II) and the compounds of the formula (III), R 1、R2、R3、R4、X、L1 and m have the same meanings as in the compounds of the formula (I).
5. The process according to claim 4, wherein when X is nitrogen and m=1 and r 1、R2 is independently selected from C 10~14 alkyl, the process for preparing the compound of formula (iii) comprises the steps of: in acetonitrile, under the action of potassium carbonate, N-tert-butoxycarbonyl-1, 2-ethylenediamine and bromoalkane react to obtain an intermediate a; reacting a dichloromethane solution of the intermediate a under the action of trifluoroacetic acid to obtain an intermediate b, namely a compound shown in a formula (III);
the specific process is as follows:
Preferably, the volume ratio of the mole amount of the N-t-butoxycarbonyl-1, 2-ethylenediamine to the acetonitrile is 0.01-1 mole/L; the molar ratio of the potassium carbonate to the N-tert-butoxycarbonyl-1, 2-ethylenediamine is 1-3:1; the mol ratio of the N-tert-butyloxycarbonyl-1, 2-ethylenediamine to the bromoalkane is 1:1-1.5; the reaction temperature of the N-tert-butoxycarbonyl-1, 2-ethylenediamine and bromoalkane is 60-100 ℃ and the reaction time is 60-80 h; the molar ratio of the intermediate a to the trifluoroacetic acid is 1-3:1; the reaction temperature of the intermediate a for producing the intermediate b is ice bath, and the reaction time is 2-8 h.
6. The process according to claim 4, wherein the organic solvent is one or more selected from the group consisting of methanol, ethanol, isopropanol, benzene, toluene, xylene, pentane, hexane, octane, cyclohexane, cyclohexanone, toluene cyclohexanone, chlorobenzene, dichlorobenzene, methylene chloride, diethyl ether, propylene oxide, acetone, methyl butanone, methyl isobutyl ketone, acetonitrile, pyridine, phenol, styrene, perchloroethylene, trichloroethylene, ethylene glycol ether, dimethyl sulfoxide, N-dimethylformamide and triethanolamine; preferably, when the solvent is methanol, dichloromethane, acetonitrile, ethyl acetate, dimethyl sulfoxide; the volume ratio of the compound shown in the formula (II) to the organic solvent is 0.01-10 mol/L;
Or, the catalyst is selected from one or more of N-hydroxysuccinimide (NHS), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI), 1-Hydroxybenzotriazole (HOBT), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea Hexafluorophosphate (HATU), O-benzotriazol-tetramethylurea Hexafluorophosphate (HBTU), 4-Dimethylaminopyridine (DMAP), N, N-Diisopropylethylamine (DIPEA) or O-benzotriazol-N-, N, N ', N' -tetramethylurea tetrafluoroboric acid (TBTU); preferably, when the catalyst is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI), 1-Hydroxybenzotriazole (HOBT) and N, N-Diisopropylethylamine (DIPEA), the molar ratio of the catalyst to the compound represented by formula (II) is 1-1.5:1;
or the mol ratio of the compound shown in the formula (II) to the compound shown in the formula (III) is 1:1-1.5; preferably 1:1.2.
Or the reaction temperature is room temperature, and the reaction time is 10-30 hours;
or, the post-treatment method of the reaction liquid obtained by reacting the compound shown in the formula (II) and the compound shown in the formula (III) comprises the following steps:
adding saturated sodium chloride solution and saturated sodium bicarbonate solution into the reaction solution, extracting with dichloromethane, collecting an organic phase, drying the organic phase by anhydrous sodium sulfate, filtering, evaporating under reduced pressure, and separating by silica gel column chromatography to obtain lipid; the eluent used for the silica gel column chromatography is a mixture of dichloromethane and methanol, DCM/meoh=200:0 to 10:1.
7. Use of an endogenous amino acid-derived ionizable lipid of claim 1, or an N-oxide, solvate, pharmaceutically acceptable salt or stereoisomer thereof, as a drug delivery vehicle.
8. A lipid nanoparticle composition comprising the endogenous amino acid-derived ionizable lipid of claim 1, a helper lipid, cholesterol, a PEG lipid, and a entrapped drug.
9. The lipid nanoparticle composition of claim 8, wherein the helper lipid is selected from a variety of sources, including distearoyl phosphatidylcholine (DSPC), dioleoyl phosphatidylcholine (DOPC), dipalmitoyl phosphatidylcholine (DPPC), dioleoyl phosphatidylglycerol (DOPG), dipalmitoyl phosphatidylglycerol (DPPG), dioleoyl phosphatidylethanolamine (DOPE), palmitoyl phosphatidylcholine (POPC), 1-palmitoyl-2-oleoyl phosphatidylethanolamine (POPE), dioleoyl phosphatidylethanolamine-maleimide (DOPE-MAL), dipalmitoyl phosphatidylethanolamine (DPPE), dipyrimidinylphosphatidylcholine (DMPE), distearoyl phosphatidylethanolamine (DSPE), and 1-stearoyl-2-oleoyl phosphatidylethanolamine (SOPE), preferably dioleoyl phosphatidylethanolamine (DOPE);
or, the cholesterol is selected from one or two of cholesterol or 10 alpha-hydroxy cholesterol; preferably, the cholesterol is cholesterol or a mixture of cholesterol and 10 alpha-hydroxy cholesterol (mass ratio is 1:1);
or, the PEG lipid is selected from polyethylene glycol-dimyristoyl glycerol (PEG-DMG), polyethylene glycol-distearoyl phosphatidylethanolamine (PEG-DSPE), polyethylene glycol-dimethacrylate (PEG-DMA); preferably polyethylene glycol-dimyristoyl glycerol (PEG-DMG);
Or, the encapsulated drug comprises one or more of biological drugs or chemical drugs; the biological medicine comprises one or more of nucleic acid medicine, protein medicine, polypeptide medicine or polysaccharide medicine; preferably, the nucleic acid drug comprises one or more of messenger RNA (mRNA), small interfering RNA (siRNA), micro RNA (microRNA), circular RNA (circRNA), long non-coding RNA (lncRNA), plasmid DNA (plasmid DNA) and small circle DNA (mini circle DNA); the chemical medicine comprises one or more of small molecule medicine, fluorescein or developer; further preferred, the nucleic acid agent is mRNA;
Or, the lipid nanoparticle composition has a diameter of 1nm to 1000nm; preferably 100 to 200nm;
or, the molar ratio of the endogenous amino acid derived ionizable lipid, the auxiliary lipid, the cholesterol and the PEG lipid is 15-45:20-35:10-40:0.5-5;
the endogenous amino acid-derived ionizable lipid of claim 1 in a mass ratio of 1-100:1 to the encapsulated drug.
10. Use of the lipid nanoparticle composition of claim 9 in the preparation of a genetic medicament comprising an active ingredient which is a nucleic acid medicament and a delivery vehicle which is the lipid nanoparticle composition of claim 9.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202410477849.1A CN118388389A (en) | 2024-04-19 | 2024-04-19 | Endogenous amino acid derived ionizable lipid, and preparation method and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202410477849.1A CN118388389A (en) | 2024-04-19 | 2024-04-19 | Endogenous amino acid derived ionizable lipid, and preparation method and application thereof |
Publications (1)
Publication Number | Publication Date |
---|---|
CN118388389A true CN118388389A (en) | 2024-07-26 |
Family
ID=91984080
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202410477849.1A Pending CN118388389A (en) | 2024-04-19 | 2024-04-19 | Endogenous amino acid derived ionizable lipid, and preparation method and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN118388389A (en) |
-
2024
- 2024-04-19 CN CN202410477849.1A patent/CN118388389A/en active Pending
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2022166213A1 (en) | Ionizable lipid molecule, preparation method therefor, and application thereof in preparation of lipid nanoparticle | |
EP4282855A1 (en) | Ionizable lipid molecule, preparation method therefor, and application thereof in preparation of lipid nanoparticle | |
CA2800401C (en) | Biodegradable lipids for the delivery of active agents | |
CN114773217B (en) | Cationic lipid compounds and compositions for delivery of nucleic acids and uses | |
WO2023134325A1 (en) | Lipid compound, composition containing same, and use | |
WO2008137470A1 (en) | Multi-chain lipophilic polyamines | |
CA3215389C (en) | Ionizable lipids and compositions for nucleic acid delivery | |
CN116199646B (en) | Tris-based ionizable lipid, and preparation method and application thereof | |
KR20220058578A (en) | Asialoglycoprotein Receptor Mediated Delivery of Therapeutically Active Conjugates | |
WO2023236976A1 (en) | Lipid compound and preparation method therefor, and use thereof | |
CN114716355A (en) | Lipid compound, composition containing same and application | |
WO2022235935A2 (en) | Ionizable cationic lipids for rna delivery | |
CN116969850A (en) | Carbonate-carboxylate type lipid compound, lipid carrier based on same, nucleic acid lipid nanoparticle composition and pharmaceutical preparation | |
CN115286674A (en) | Cationic liposome and preparation method thereof | |
CN117623978A (en) | Biodegradable amino acid derived ionizable lipid, and preparation method and application thereof | |
JPWO2016027699A1 (en) | Cationic lipids for nucleic acid delivery | |
CN118388389A (en) | Endogenous amino acid derived ionizable lipid, and preparation method and application thereof | |
EP3456714B1 (en) | Biodegradable compound, lipid particle, composition containing lipid particle and kit | |
CN114539083B (en) | Lipid nanoparticles and their use in nucleic acid delivery | |
CN114181200A (en) | Cationic liposome with high gene transfection efficiency and preparation and application thereof | |
CN116143707B (en) | Base ionizable lipid and preparation method and application thereof | |
US20030065033A1 (en) | Lipid derivatives of polythiourea | |
CN118084714B (en) | Lipid compound, lipid nanoparticle containing lipid compound and application of lipid compound | |
CN118125932A (en) | Application of ionizable lipid molecules and nanoparticle composition thereof in nucleic acid delivery | |
CN117534585A (en) | Novel ionizable cationic lipid compound, and preparation method and application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination |