CN110114058A - 用于递送mrna的改进的基于ice的脂质纳米颗粒制剂 - Google Patents
用于递送mrna的改进的基于ice的脂质纳米颗粒制剂 Download PDFInfo
- Publication number
- CN110114058A CN110114058A CN201780080493.6A CN201780080493A CN110114058A CN 110114058 A CN110114058 A CN 110114058A CN 201780080493 A CN201780080493 A CN 201780080493A CN 110114058 A CN110114058 A CN 110114058A
- Authority
- CN
- China
- Prior art keywords
- mrna
- lipid
- sterol
- composition
- composition according
- 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.)
- Granted
Links
- 150000002632 lipids Chemical class 0.000 title claims abstract description 188
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 155
- 108020004999 messenger RNA Proteins 0.000 title claims description 459
- 238000002360 preparation method Methods 0.000 title abstract description 91
- 239000000203 mixture Substances 0.000 claims abstract description 236
- -1 cation lipid Chemical class 0.000 claims abstract description 219
- 229930182558 Sterol Natural products 0.000 claims abstract description 134
- 235000003702 sterols Nutrition 0.000 claims abstract description 134
- 238000000034 method Methods 0.000 claims abstract description 120
- 150000003432 sterols Chemical class 0.000 claims abstract description 119
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 87
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 83
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 83
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 158
- 235000019441 ethanol Nutrition 0.000 claims description 83
- 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 78
- 108090000623 proteins and genes Proteins 0.000 claims description 75
- 102000004169 proteins and genes Human genes 0.000 claims description 74
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 51
- 230000004048 modification Effects 0.000 claims description 48
- 238000012986 modification Methods 0.000 claims description 48
- 235000012000 cholesterol Nutrition 0.000 claims description 40
- 230000009467 reduction Effects 0.000 claims description 39
- OILXMJHPFNGGTO-UHFFFAOYSA-N (22E)-(24xi)-24-methylcholesta-5,22-dien-3beta-ol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(C)C(C)C)C1(C)CC2 OILXMJHPFNGGTO-UHFFFAOYSA-N 0.000 claims description 34
- OQMZNAMGEHIHNN-UHFFFAOYSA-N 7-Dehydrostigmasterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(CC)C(C)C)CCC33)C)C3=CC=C21 OQMZNAMGEHIHNN-UHFFFAOYSA-N 0.000 claims description 34
- 125000000217 alkyl group Chemical group 0.000 claims description 34
- 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 34
- 125000003729 nucleotide group Chemical group 0.000 claims description 31
- 238000012384 transportation and delivery Methods 0.000 claims description 31
- 239000002773 nucleotide Substances 0.000 claims description 24
- 230000002685 pulmonary effect Effects 0.000 claims description 23
- 125000005647 linker group Chemical group 0.000 claims description 22
- 238000005457 optimization Methods 0.000 claims description 22
- 108020004705 Codon Proteins 0.000 claims description 21
- 230000003647 oxidation Effects 0.000 claims description 20
- 238000007254 oxidation reaction Methods 0.000 claims description 20
- 238000001727 in vivo Methods 0.000 claims description 19
- 238000005507 spraying Methods 0.000 claims description 18
- XSQUKJJJFZCRTK-UHFFFAOYSA-N urea group Chemical group NC(=O)N XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 18
- KZJWDPNRJALLNS-VPUBHVLGSA-N (-)-beta-Sitosterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]([C@H](CC[C@@H](C(C)C)CC)C)CC4)CC3)CC=2)CC1 KZJWDPNRJALLNS-VPUBHVLGSA-N 0.000 claims description 17
- CSVWWLUMXNHWSU-UHFFFAOYSA-N (22E)-(24xi)-24-ethyl-5alpha-cholest-22-en-3beta-ol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(CC)C(C)C)C1(C)CC2 CSVWWLUMXNHWSU-UHFFFAOYSA-N 0.000 claims description 17
- RQOCXCFLRBRBCS-UHFFFAOYSA-N (22E)-cholesta-5,7,22-trien-3beta-ol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CCC(C)C)CCC33)C)C3=CC=C21 RQOCXCFLRBRBCS-UHFFFAOYSA-N 0.000 claims description 17
- KLEXDBGYSOIREE-UHFFFAOYSA-N 24xi-n-propylcholesterol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CCC)C(C)C)C1(C)CC2 KLEXDBGYSOIREE-UHFFFAOYSA-N 0.000 claims description 17
- LPZCCMIISIBREI-MTFRKTCUSA-N Citrostadienol Natural products CC=C(CC[C@@H](C)[C@H]1CC[C@H]2C3=CC[C@H]4[C@H](C)[C@@H](O)CC[C@]4(C)[C@H]3CC[C@]12C)C(C)C LPZCCMIISIBREI-MTFRKTCUSA-N 0.000 claims description 17
- ARVGMISWLZPBCH-UHFFFAOYSA-N Dehydro-beta-sitosterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)CCC(CC)C(C)C)CCC33)C)C3=CC=C21 ARVGMISWLZPBCH-UHFFFAOYSA-N 0.000 claims description 17
- DNVPQKQSNYMLRS-NXVQYWJNSA-N Ergosterol Natural products CC(C)[C@@H](C)C=C[C@H](C)[C@H]1CC[C@H]2C3=CC=C4C[C@@H](O)CC[C@]4(C)[C@@H]3CC[C@]12C DNVPQKQSNYMLRS-NXVQYWJNSA-N 0.000 claims description 17
- HZYXFRGVBOPPNZ-UHFFFAOYSA-N UNPD88870 Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)=CCC(CC)C(C)C)C1(C)CC2 HZYXFRGVBOPPNZ-UHFFFAOYSA-N 0.000 claims description 17
- MJVXAPPOFPTTCA-UHFFFAOYSA-N beta-Sistosterol Natural products CCC(CCC(C)C1CCC2C3CC=C4C(C)C(O)CCC4(C)C3CCC12C)C(C)C MJVXAPPOFPTTCA-UHFFFAOYSA-N 0.000 claims description 17
- 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 17
- DNVPQKQSNYMLRS-SOWFXMKYSA-N ergosterol Chemical compound C1[C@@H](O)CC[C@]2(C)[C@H](CC[C@]3([C@H]([C@H](C)/C=C/[C@@H](C)C(C)C)CC[C@H]33)C)C3=CC=C21 DNVPQKQSNYMLRS-SOWFXMKYSA-N 0.000 claims description 17
- 235000015500 sitosterol Nutrition 0.000 claims description 17
- 229950005143 sitosterol Drugs 0.000 claims description 17
- NLQLSVXGSXCXFE-UHFFFAOYSA-N sitosterol Natural products CC=C(/CCC(C)C1CC2C3=CCC4C(C)C(O)CCC4(C)C3CCC2(C)C1)C(C)C NLQLSVXGSXCXFE-UHFFFAOYSA-N 0.000 claims description 17
- 229940032091 stigmasterol Drugs 0.000 claims description 17
- HCXVJBMSMIARIN-PHZDYDNGSA-N stigmasterol 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)/C=C/[C@@H](CC)C(C)C)[C@@]1(C)CC2 HCXVJBMSMIARIN-PHZDYDNGSA-N 0.000 claims description 17
- 235000016831 stigmasterol Nutrition 0.000 claims description 17
- BFDNMXAIBMJLBB-UHFFFAOYSA-N stigmasterol Natural products CCC(C=CC(C)C1CCCC2C3CC=C4CC(O)CCC4(C)C3CCC12C)C(C)C BFDNMXAIBMJLBB-UHFFFAOYSA-N 0.000 claims description 17
- 125000003368 amide group Chemical group 0.000 claims description 16
- GKBHKNPLNHLYHT-UHFFFAOYSA-N 5beta-Stigmastan Natural products C1CC2CCCCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 GKBHKNPLNHLYHT-UHFFFAOYSA-N 0.000 claims description 15
- SGNBVLSWZMBQTH-FGAXOLDCSA-N Campesterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]([C@H](CC[C@H](C(C)C)C)C)CC4)CC3)CC=2)CC1 SGNBVLSWZMBQTH-FGAXOLDCSA-N 0.000 claims description 15
- BTEISVKTSQLKST-UHFFFAOYSA-N Haliclonasterol Natural products CC(C=CC(C)C(C)(C)C)C1CCC2C3=CC=C4CC(O)CCC4(C)C3CCC12C BTEISVKTSQLKST-UHFFFAOYSA-N 0.000 claims description 15
- 230000015572 biosynthetic process Effects 0.000 claims description 15
- SGNBVLSWZMBQTH-PODYLUTMSA-N campesterol 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](C)C(C)C)[C@@]1(C)CC2 SGNBVLSWZMBQTH-PODYLUTMSA-N 0.000 claims description 15
- 235000000431 campesterol Nutrition 0.000 claims description 15
- 125000003226 pyrazolyl group Chemical group 0.000 claims description 15
- GKBHKNPLNHLYHT-LWQAOISPSA-N stigmastane Chemical compound C1CC2CCCC[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 GKBHKNPLNHLYHT-LWQAOISPSA-N 0.000 claims description 15
- LAGFCVRVICMFFF-UHFFFAOYSA-N stigmastane Natural products CCC(CCC(C)C1CCC2C3CCC4CC(=O)CC(C)C4C3CCC12C)C(C)C LAGFCVRVICMFFF-UHFFFAOYSA-N 0.000 claims description 15
- 125000001425 triazolyl group Chemical group 0.000 claims description 15
- 125000003373 pyrazinyl group Chemical group 0.000 claims description 14
- 125000002098 pyridazinyl group Chemical group 0.000 claims description 14
- KAESVJOAVNADME-UHFFFAOYSA-N 1H-pyrrole Natural products C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 claims description 13
- 125000005587 carbonate group Chemical group 0.000 claims description 13
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 claims description 13
- 229910052799 carbon Inorganic materials 0.000 claims description 12
- 108091033319 polynucleotide Proteins 0.000 claims description 12
- 102000040430 polynucleotide Human genes 0.000 claims description 12
- 239000002157 polynucleotide Substances 0.000 claims description 12
- 238000003786 synthesis reaction Methods 0.000 claims description 12
- 108020004414 DNA Proteins 0.000 claims description 11
- KXDHJXZQYSOELW-UHFFFAOYSA-N Carbamic acid Chemical group NC(O)=O KXDHJXZQYSOELW-UHFFFAOYSA-N 0.000 claims description 10
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 claims description 10
- 125000004076 pyridyl group Chemical group 0.000 claims description 9
- 150000002460 imidazoles Chemical class 0.000 claims description 7
- 101150077194 CAP1 gene Proteins 0.000 claims description 6
- 201000003883 Cystic fibrosis Diseases 0.000 claims description 6
- 101100245221 Mus musculus Prss8 gene Proteins 0.000 claims description 6
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical group C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 claims description 6
- 108020004459 Small interfering RNA Proteins 0.000 claims description 6
- 125000004185 ester group Chemical group 0.000 claims description 6
- 238000006467 substitution reaction Methods 0.000 claims description 6
- 239000001257 hydrogen Substances 0.000 claims description 5
- 229910052739 hydrogen Inorganic materials 0.000 claims description 5
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 4
- 108700011259 MicroRNAs Proteins 0.000 claims description 4
- 239000004020 conductor Substances 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 239000002679 microRNA Substances 0.000 claims description 4
- 239000004202 carbamide Substances 0.000 claims description 3
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical group C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 claims 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Chemical group COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims 1
- 125000003831 tetrazolyl group Chemical group 0.000 claims 1
- 210000004072 lung Anatomy 0.000 description 155
- 102000012605 Cystic Fibrosis Transmembrane Conductance Regulator Human genes 0.000 description 88
- 108010079245 Cystic Fibrosis Transmembrane Conductance Regulator Proteins 0.000 description 88
- 241000699666 Mus <mouse, genus> Species 0.000 description 74
- 239000002502 liposome Substances 0.000 description 69
- 210000001519 tissue Anatomy 0.000 description 68
- 239000002585 base Substances 0.000 description 54
- 241000700159 Rattus Species 0.000 description 47
- 229920001223 polyethylene glycol Polymers 0.000 description 43
- 239000002202 Polyethylene glycol Substances 0.000 description 40
- 238000012545 processing Methods 0.000 description 39
- 150000003838 adenosines Chemical class 0.000 description 37
- 229930183912 Cytidylic acid Natural products 0.000 description 36
- IERHLVCPSMICTF-XVFCMESISA-N cytidine 5'-monophosphate Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(O)=O)O1 IERHLVCPSMICTF-XVFCMESISA-N 0.000 description 36
- IERHLVCPSMICTF-UHFFFAOYSA-N cytidine monophosphate Natural products O=C1N=C(N)C=CN1C1C(O)C(O)C(COP(O)(O)=O)O1 IERHLVCPSMICTF-UHFFFAOYSA-N 0.000 description 36
- 239000000243 solution Substances 0.000 description 36
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 31
- 238000004090 dissolution Methods 0.000 description 29
- 150000001413 amino acids Chemical class 0.000 description 28
- 230000008569 process Effects 0.000 description 28
- 229940048102 triphosphoric acid Drugs 0.000 description 28
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 27
- 229940024606 amino acid Drugs 0.000 description 27
- 241001465754 Metazoa Species 0.000 description 26
- 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 description 25
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 22
- 238000002156 mixing Methods 0.000 description 22
- 239000000443 aerosol Substances 0.000 description 21
- 239000007979 citrate buffer Substances 0.000 description 21
- 201000010099 disease Diseases 0.000 description 21
- 239000000126 substance Substances 0.000 description 20
- 210000004027 cell Anatomy 0.000 description 19
- 238000009472 formulation Methods 0.000 description 19
- 230000003053 immunization Effects 0.000 description 18
- 238000002649 immunization Methods 0.000 description 18
- 229910052757 nitrogen Inorganic materials 0.000 description 18
- 238000004458 analytical method Methods 0.000 description 17
- 238000004043 dyeing Methods 0.000 description 17
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 16
- 238000005086 pumping Methods 0.000 description 16
- 239000011780 sodium chloride Substances 0.000 description 16
- 239000000872 buffer Substances 0.000 description 15
- 150000003839 salts Chemical class 0.000 description 15
- 239000011550 stock solution Substances 0.000 description 15
- 235000005979 Citrus limon Nutrition 0.000 description 14
- 244000131522 Citrus pyriformis Species 0.000 description 14
- 239000002245 particle Substances 0.000 description 13
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 description 12
- 238000009826 distribution Methods 0.000 description 12
- 239000002777 nucleoside Substances 0.000 description 12
- 239000002253 acid Substances 0.000 description 11
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 11
- 210000000424 bronchial epithelial cell Anatomy 0.000 description 11
- 108091034117 Oligonucleotide Proteins 0.000 description 10
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 10
- 238000011532 immunohistochemical staining Methods 0.000 description 10
- 238000012360 testing method Methods 0.000 description 10
- 241000282414 Homo sapiens Species 0.000 description 9
- 210000002966 serum Anatomy 0.000 description 9
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 108090000331 Firefly luciferases Proteins 0.000 description 8
- 229910019142 PO4 Inorganic materials 0.000 description 8
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 8
- 238000007385 chemical modification Methods 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 229930182470 glycoside Natural products 0.000 description 8
- 150000002338 glycosides Chemical class 0.000 description 8
- 239000003921 oil Substances 0.000 description 8
- 239000012188 paraffin wax Substances 0.000 description 8
- 239000008194 pharmaceutical composition Substances 0.000 description 8
- 239000010452 phosphate Substances 0.000 description 8
- 239000007787 solid Substances 0.000 description 8
- 238000012546 transfer Methods 0.000 description 8
- ZDTFMPXQUSBYRL-UUOKFMHZSA-N 2-Aminoadenosine Chemical compound C12=NC(N)=NC(N)=C2N=CN1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O ZDTFMPXQUSBYRL-UUOKFMHZSA-N 0.000 description 7
- 241000699670 Mus sp. Species 0.000 description 7
- 238000005411 Van der Waals force Methods 0.000 description 7
- 238000000889 atomisation Methods 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 239000007789 gas Substances 0.000 description 7
- 238000003306 harvesting Methods 0.000 description 7
- XNGIFLGASWRNHJ-UHFFFAOYSA-L phthalate(2-) Chemical compound [O-]C(=O)C1=CC=CC=C1C([O-])=O XNGIFLGASWRNHJ-UHFFFAOYSA-L 0.000 description 7
- 102000004196 processed proteins & peptides Human genes 0.000 description 7
- 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 description 7
- 229940035893 uracil Drugs 0.000 description 7
- ZAYHVCMSTBRABG-JXOAFFINSA-N 5-methylcytidine Chemical compound O=C1N=C(N)C(C)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 ZAYHVCMSTBRABG-JXOAFFINSA-N 0.000 description 6
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 6
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 description 6
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 description 6
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 6
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 6
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 6
- 125000002252 acyl group Chemical group 0.000 description 6
- 229960005305 adenosine Drugs 0.000 description 6
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 6
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 239000003795 chemical substances by application Substances 0.000 description 6
- 238000009295 crossflow filtration Methods 0.000 description 6
- 230000034994 death Effects 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 229940029575 guanosine Drugs 0.000 description 6
- 239000002071 nanotube Substances 0.000 description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 6
- 229940045145 uridine Drugs 0.000 description 6
- UHDGCWIWMRVCDJ-UHFFFAOYSA-N 1-beta-D-Xylofuranosyl-NH-Cytosine Natural products O=C1N=C(N)C=CN1C1C(O)C(O)C(CO)O1 UHDGCWIWMRVCDJ-UHFFFAOYSA-N 0.000 description 5
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 5
- 108020003589 5' Untranslated Regions Proteins 0.000 description 5
- 241000282472 Canis lupus familiaris Species 0.000 description 5
- UHDGCWIWMRVCDJ-PSQAKQOGSA-N Cytidine Natural products O=C1N=C(N)C=CN1[C@@H]1[C@@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-PSQAKQOGSA-N 0.000 description 5
- 229930010555 Inosine Natural products 0.000 description 5
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 5
- 241000124008 Mammalia Species 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- UHDGCWIWMRVCDJ-ZAKLUEHWSA-N cytidine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-ZAKLUEHWSA-N 0.000 description 5
- 238000007710 freezing Methods 0.000 description 5
- 230000008014 freezing Effects 0.000 description 5
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 5
- 229960003786 inosine Drugs 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 5
- 229920001184 polypeptide Polymers 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 230000028327 secretion Effects 0.000 description 5
- 239000008279 sol Substances 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 235000000346 sugar Nutrition 0.000 description 5
- 231100000419 toxicity Toxicity 0.000 description 5
- 230000001988 toxicity Effects 0.000 description 5
- 108020005345 3' Untranslated Regions Proteins 0.000 description 4
- ZAYHVCMSTBRABG-UHFFFAOYSA-N 5-Methylcytidine Natural products O=C1N=C(N)C(C)=CN1C1C(O)C(O)C(CO)O1 ZAYHVCMSTBRABG-UHFFFAOYSA-N 0.000 description 4
- 108091027075 5S-rRNA precursor Proteins 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- 101710163270 Nuclease Proteins 0.000 description 4
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 4
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 description 4
- 150000001412 amines Chemical class 0.000 description 4
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 4
- 238000010276 construction Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000013265 extended release Methods 0.000 description 4
- 230000006872 improvement Effects 0.000 description 4
- GLGLUQVVDHRLQK-WRBBJXAJSA-N n,n-dimethyl-2,3-bis[(z)-octadec-9-enoxy]propan-1-amine Chemical compound CCCCCCCC\C=C/CCCCCCCCOCC(CN(C)C)OCCCCCCCC\C=C/CCCCCCCC GLGLUQVVDHRLQK-WRBBJXAJSA-N 0.000 description 4
- 230000007935 neutral effect Effects 0.000 description 4
- 150000003833 nucleoside derivatives Chemical class 0.000 description 4
- 125000003835 nucleoside group Chemical group 0.000 description 4
- 229940055577 oleyl alcohol Drugs 0.000 description 4
- XMLQWXUVTXCDDL-UHFFFAOYSA-N oleyl alcohol Natural products CCCCCCC=CCCCCCCCCCCO XMLQWXUVTXCDDL-UHFFFAOYSA-N 0.000 description 4
- 125000001820 oxy group Chemical group [*:1]O[*:2] 0.000 description 4
- 210000002706 plastid Anatomy 0.000 description 4
- 239000001294 propane Substances 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 description 3
- 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 description 3
- LDGWQMRUWMSZIU-LQDDAWAPSA-M 2,3-bis[(z)-octadec-9-enoxy]propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCC\C=C/CCCCCCCCOCC(C[N+](C)(C)C)OCCCCCCCC\C=C/CCCCCCCC LDGWQMRUWMSZIU-LQDDAWAPSA-M 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 3
- WOVKYSAHUYNSMH-UHFFFAOYSA-N BROMODEOXYURIDINE Natural products C1C(O)C(CO)OC1N1C(=O)NC(=O)C(Br)=C1 WOVKYSAHUYNSMH-UHFFFAOYSA-N 0.000 description 3
- 239000004380 Cholic acid Substances 0.000 description 3
- 241000283073 Equus caballus Species 0.000 description 3
- XKMLYUALXHKNFT-UUOKFMHZSA-N Guanosine-5'-triphosphate Chemical compound C1=2NC(N)=NC(=O)C=2N=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XKMLYUALXHKNFT-UUOKFMHZSA-N 0.000 description 3
- 241000880493 Leptailurus serval Species 0.000 description 3
- 241000283973 Oryctolagus cuniculus Species 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 241001494479 Pecora Species 0.000 description 3
- UNBFGVQVQGXXCK-KKUMJFAQSA-N Phe-Ser-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O UNBFGVQVQGXXCK-KKUMJFAQSA-N 0.000 description 3
- 239000004698 Polyethylene Substances 0.000 description 3
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 3
- 108091034057 RNA (poly(A)) Proteins 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 3
- 241000282898 Sus scrofa Species 0.000 description 3
- 241000218636 Thuja Species 0.000 description 3
- 125000000539 amino acid group Chemical group 0.000 description 3
- 239000012062 aqueous buffer Substances 0.000 description 3
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 3
- 108010062796 arginyllysine Proteins 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 3
- 229950004398 broxuridine Drugs 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 210000004899 c-terminal region Anatomy 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 235000019416 cholic acid Nutrition 0.000 description 3
- 229960002471 cholic acid Drugs 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000005034 decoration Methods 0.000 description 3
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000011026 diafiltration Methods 0.000 description 3
- 231100000673 dose–response relationship Toxicity 0.000 description 3
- 235000013601 eggs Nutrition 0.000 description 3
- 210000002919 epithelial cell Anatomy 0.000 description 3
- 229940031098 ethanolamine Drugs 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 229910052731 fluorine Inorganic materials 0.000 description 3
- 239000011737 fluorine Substances 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 108020001507 fusion proteins Proteins 0.000 description 3
- 102000037865 fusion proteins Human genes 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 229910052740 iodine Inorganic materials 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 125000003473 lipid group Chemical group 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 108010045758 lysosomal proteins Proteins 0.000 description 3
- 210000003712 lysosome Anatomy 0.000 description 3
- 230000001868 lysosomic effect Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- QEWYKACRFQMRMB-UHFFFAOYSA-N monofluoroacetic acid Natural products OC(=O)CF QEWYKACRFQMRMB-UHFFFAOYSA-N 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 230000002093 peripheral effect Effects 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- YNPNZTXNASCQKK-UHFFFAOYSA-N phenanthrene Chemical compound C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 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 description 3
- 239000011148 porous material Substances 0.000 description 3
- 239000002719 pyrimidine nucleotide Substances 0.000 description 3
- 150000003230 pyrimidines Chemical class 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000036387 respiratory rate Effects 0.000 description 3
- 239000012465 retentate Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 230000002194 synthesizing effect Effects 0.000 description 3
- 238000012385 systemic delivery Methods 0.000 description 3
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 3
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 238000013519 translation Methods 0.000 description 3
- UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical group N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- RIFDKYBNWNPCQK-IOSLPCCCSA-N (2r,3s,4r,5r)-2-(hydroxymethyl)-5-(6-imino-3-methylpurin-9-yl)oxolane-3,4-diol Chemical compound C1=2N(C)C=NC(=N)C=2N=CN1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O RIFDKYBNWNPCQK-IOSLPCCCSA-N 0.000 description 2
- 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 description 2
- PISWNSOQFZRVJK-XLPZGREQSA-N 1-[(2r,4s,5r)-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-5-methyl-2-sulfanylidenepyrimidin-4-one Chemical compound S=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 PISWNSOQFZRVJK-XLPZGREQSA-N 0.000 description 2
- HPZMWTNATZPBIH-UHFFFAOYSA-N 1-methyladenine Chemical compound CN1C=NC2=NC=NC2=C1N HPZMWTNATZPBIH-UHFFFAOYSA-N 0.000 description 2
- RFLVMTUMFYRZCB-UHFFFAOYSA-N 1-methylguanine Chemical compound O=C1N(C)C(N)=NC2=C1N=CN2 RFLVMTUMFYRZCB-UHFFFAOYSA-N 0.000 description 2
- XQCZBXHVTFVIFE-UHFFFAOYSA-N 2-amino-4-hydroxypyrimidine Chemical compound NC1=NC=CC(O)=N1 XQCZBXHVTFVIFE-UHFFFAOYSA-N 0.000 description 2
- FZWGECJQACGGTI-UHFFFAOYSA-N 2-amino-7-methyl-1,7-dihydro-6H-purin-6-one Chemical compound NC1=NC(O)=C2N(C)C=NC2=N1 FZWGECJQACGGTI-UHFFFAOYSA-N 0.000 description 2
- ZLOIGESWDJYCTF-XVFCMESISA-N 4-thiouridine Chemical group O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=S)C=C1 ZLOIGESWDJYCTF-XVFCMESISA-N 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 2
- OIVLITBTBDPEFK-UHFFFAOYSA-N 5,6-dihydrouracil Chemical compound O=C1CCNC(=O)N1 OIVLITBTBDPEFK-UHFFFAOYSA-N 0.000 description 2
- LQLQRFGHAALLLE-UHFFFAOYSA-N 5-bromouracil Chemical compound BrC1=CNC(=O)NC1=O LQLQRFGHAALLLE-UHFFFAOYSA-N 0.000 description 2
- LRSASMSXMSNRBT-UHFFFAOYSA-N 5-methylcytosine Chemical compound CC1=CNC(=O)N=C1N LRSASMSXMSNRBT-UHFFFAOYSA-N 0.000 description 2
- KDOPAZIWBAHVJB-UHFFFAOYSA-N 5h-pyrrolo[3,2-d]pyrimidine Chemical compound C1=NC=C2NC=CC2=N1 KDOPAZIWBAHVJB-UHFFFAOYSA-N 0.000 description 2
- BXJHWYVXLGLDMZ-UHFFFAOYSA-N 6-O-methylguanine Chemical compound COC1=NC(N)=NC2=C1NC=N2 BXJHWYVXLGLDMZ-UHFFFAOYSA-N 0.000 description 2
- UEHOMUNTZPIBIL-UUOKFMHZSA-N 6-amino-9-[(2r,3r,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-7h-purin-8-one Chemical compound O=C1NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O UEHOMUNTZPIBIL-UUOKFMHZSA-N 0.000 description 2
- HCAJQHYUCKICQH-VPENINKCSA-N 8-Oxo-7,8-dihydro-2'-deoxyguanosine Chemical compound C1=2NC(N)=NC(=O)C=2NC(=O)N1[C@H]1C[C@H](O)[C@@H](CO)O1 HCAJQHYUCKICQH-VPENINKCSA-N 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 2
- 108010085238 Actins Proteins 0.000 description 2
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 2
- OMSKGWFGWCQFBD-KZVJFYERSA-N Ala-Val-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OMSKGWFGWCQFBD-KZVJFYERSA-N 0.000 description 2
- 108091029845 Aminoallyl nucleotide Proteins 0.000 description 2
- 102000053640 Argininosuccinate synthases Human genes 0.000 description 2
- 108700024106 Argininosuccinate synthases Proteins 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 2
- YDNKGFDKKRUKPY-JHOUSYSJSA-N C16 ceramide Natural products CCCCCCCCCCCCCCCC(=O)N[C@@H](CO)[C@H](O)C=CCCCCCCCCCCCCC YDNKGFDKKRUKPY-JHOUSYSJSA-N 0.000 description 2
- 101150014715 CAP2 gene Proteins 0.000 description 2
- 108091079001 CRISPR RNA Proteins 0.000 description 2
- 102000011045 Chloride Channels Human genes 0.000 description 2
- 108010062745 Chloride Channels Proteins 0.000 description 2
- PCDQPRRSZKQHHS-UHFFFAOYSA-N Cytidine 5'-triphosphate Natural products O=C1N=C(N)C=CN1C1C(O)C(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 PCDQPRRSZKQHHS-UHFFFAOYSA-N 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- XULFJDKZVHTRLG-JDVCJPALSA-N DOSPA trifluoroacetate Chemical compound [O-]C(=O)C(F)(F)F.CCCCCCCC\C=C/CCCCCCCCOCC(C[N+](C)(C)CCNC(=O)C(CCCNCCCN)NCCCN)OCCCCCCCC\C=C/CCCCCCCC XULFJDKZVHTRLG-JDVCJPALSA-N 0.000 description 2
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 2
- 102000002322 Egg Proteins Human genes 0.000 description 2
- 108010000912 Egg Proteins Proteins 0.000 description 2
- 108060002716 Exonuclease Proteins 0.000 description 2
- 241000282326 Felis catus Species 0.000 description 2
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 2
- QKCZZAZNMMVICF-DCAQKATOSA-N Gln-Leu-Glu Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O QKCZZAZNMMVICF-DCAQKATOSA-N 0.000 description 2
- OZEQPCDLCDRCGY-SOUVJXGZSA-N Gln-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CCC(=O)N)N)C(=O)O OZEQPCDLCDRCGY-SOUVJXGZSA-N 0.000 description 2
- HUWSBFYAGXCXKC-CIUDSAMLSA-N Glu-Ala-Met Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O HUWSBFYAGXCXKC-CIUDSAMLSA-N 0.000 description 2
- OQXDUSZKISQQSS-GUBZILKMSA-N Glu-Lys-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O OQXDUSZKISQQSS-GUBZILKMSA-N 0.000 description 2
- DTRUBYPMMVPQPD-YUMQZZPRSA-N Gly-Gln-Arg Chemical compound [H]NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O DTRUBYPMMVPQPD-YUMQZZPRSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 108020005004 Guide RNA Proteins 0.000 description 2
- JGFWUKYIQAEYAH-DCAQKATOSA-N His-Ser-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O JGFWUKYIQAEYAH-DCAQKATOSA-N 0.000 description 2
- 102000002265 Human Growth Hormone Human genes 0.000 description 2
- 108010000521 Human Growth Hormone Proteins 0.000 description 2
- 239000000854 Human Growth Hormone Substances 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- QICVAHODWHIWIS-HTFCKZLJSA-N Ile-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N QICVAHODWHIWIS-HTFCKZLJSA-N 0.000 description 2
- BEWFWZRGBDVXRP-PEFMBERDSA-N Ile-Glu-Asn Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O BEWFWZRGBDVXRP-PEFMBERDSA-N 0.000 description 2
- JNLSTRPWUXOORL-MMWGEVLESA-N Ile-Ser-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N JNLSTRPWUXOORL-MMWGEVLESA-N 0.000 description 2
- VSFNAZLYGOOSEY-UHFFFAOYSA-N Imidazolylpropionic acid Natural products OC(=O)CCN1C=CN=C1 VSFNAZLYGOOSEY-UHFFFAOYSA-N 0.000 description 2
- KGCLIYGPQXUNLO-IUCAKERBSA-N Leu-Gly-Glu Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O KGCLIYGPQXUNLO-IUCAKERBSA-N 0.000 description 2
- AOFYPTOHESIBFZ-KKUMJFAQSA-N Leu-His-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O AOFYPTOHESIBFZ-KKUMJFAQSA-N 0.000 description 2
- 101100260872 Mus musculus Tmprss4 gene Proteins 0.000 description 2
- HYVABZIGRDEKCD-UHFFFAOYSA-N N(6)-dimethylallyladenine Chemical compound CC(C)=CCNC1=NC=NC2=C1N=CN2 HYVABZIGRDEKCD-UHFFFAOYSA-N 0.000 description 2
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 2
- CRJGESKKUOMBCT-VQTJNVASSA-N N-acetylsphinganine Chemical compound CCCCCCCCCCCCCCC[C@@H](O)[C@H](CO)NC(C)=O CRJGESKKUOMBCT-VQTJNVASSA-N 0.000 description 2
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- 108091092724 Noncoding DNA Proteins 0.000 description 2
- 108020004485 Nonsense Codon Proteins 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 2
- 244000046052 Phaseolus vulgaris Species 0.000 description 2
- KNYPNEYICHHLQL-ACRUOGEOSA-N Phe-Leu-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 KNYPNEYICHHLQL-ACRUOGEOSA-N 0.000 description 2
- GPLWGAYGROGDEN-BZSNNMDCSA-N Phe-Phe-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O GPLWGAYGROGDEN-BZSNNMDCSA-N 0.000 description 2
- 108010069013 Phenylalanine Hydroxylase Proteins 0.000 description 2
- 102100038223 Phenylalanine-4-hydroxylase Human genes 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 description 2
- 241000288906 Primates Species 0.000 description 2
- 229930185560 Pseudouridine Natural products 0.000 description 2
- PTJWIQPHWPFNBW-UHFFFAOYSA-N Pseudouridine C Natural products OC1C(O)C(CO)OC1C1=CNC(=O)NC1=O PTJWIQPHWPFNBW-UHFFFAOYSA-N 0.000 description 2
- KYQCOXFCLRTKLS-UHFFFAOYSA-N Pyrazine Chemical group C1=CN=CC=N1 KYQCOXFCLRTKLS-UHFFFAOYSA-N 0.000 description 2
- 108091058545 Secretory proteins Proteins 0.000 description 2
- 102000040739 Secretory proteins Human genes 0.000 description 2
- IDQFQFVEWMWRQQ-DLOVCJGASA-N Ser-Ala-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O IDQFQFVEWMWRQQ-DLOVCJGASA-N 0.000 description 2
- PURRNJBBXDDWLX-ZDLURKLDSA-N Ser-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CO)N)O PURRNJBBXDDWLX-ZDLURKLDSA-N 0.000 description 2
- MFQMZDPAZRZAPV-NAKRPEOUSA-N Ser-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CO)N MFQMZDPAZRZAPV-NAKRPEOUSA-N 0.000 description 2
- 102000039471 Small Nuclear RNA Human genes 0.000 description 2
- 108091027967 Small hairpin RNA Proteins 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 108091046869 Telomeric non-coding RNA Proteins 0.000 description 2
- DEGCBBCMYWNJNA-RHYQMDGZSA-N Thr-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)[C@@H](C)O DEGCBBCMYWNJNA-RHYQMDGZSA-N 0.000 description 2
- AHERARIZBPOMNU-KATARQTJSA-N Thr-Ser-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O AHERARIZBPOMNU-KATARQTJSA-N 0.000 description 2
- VFJIWSJKZJTQII-SRVKXCTJSA-N Tyr-Asp-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O VFJIWSJKZJTQII-SRVKXCTJSA-N 0.000 description 2
- PGAVKCOVUIYSFO-XVFCMESISA-N UTP Chemical compound O[C@@H]1[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O[C@H]1N1C(=O)NC(=O)C=C1 PGAVKCOVUIYSFO-XVFCMESISA-N 0.000 description 2
- VXDSPJJQUQDCKH-UKJIMTQDSA-N Val-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N VXDSPJJQUQDCKH-UKJIMTQDSA-N 0.000 description 2
- HIHOWBSBBDRPDW-PTHRTHQKSA-N [(3s,8s,9s,10r,13r,14s,17r)-10,13-dimethyl-17-[(2r)-6-methylheptan-2-yl]-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthren-3-yl] n-[2-(dimethylamino)ethyl]carbamate Chemical compound C1C=C2C[C@@H](OC(=O)NCCN(C)C)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 HIHOWBSBBDRPDW-PTHRTHQKSA-N 0.000 description 2
- 108010005233 alanylglutamic acid Proteins 0.000 description 2
- 108010087924 alanylproline Proteins 0.000 description 2
- 150000001299 aldehydes Chemical group 0.000 description 2
- 229910052783 alkali metal Inorganic materials 0.000 description 2
- 238000005267 amalgamation Methods 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 2
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 2
- 108010093581 aspartyl-proline Proteins 0.000 description 2
- 125000004429 atom Chemical group 0.000 description 2
- WGQKYBSKWIADBV-UHFFFAOYSA-N benzylamine Chemical compound NCC1=CC=CC=C1 WGQKYBSKWIADBV-UHFFFAOYSA-N 0.000 description 2
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 2
- DRTQHJPVMGBUCF-PSQAKQOGSA-N beta-L-uridine Natural products O[C@H]1[C@@H](O)[C@H](CO)O[C@@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-PSQAKQOGSA-N 0.000 description 2
- WGDUUQDYDIIBKT-UHFFFAOYSA-N beta-Pseudouridine Natural products OC1OC(CN2C=CC(=O)NC2=O)C(O)C1O WGDUUQDYDIIBKT-UHFFFAOYSA-N 0.000 description 2
- UORVGPXVDQYIDP-UHFFFAOYSA-N borane Chemical compound B UORVGPXVDQYIDP-UHFFFAOYSA-N 0.000 description 2
- 239000001506 calcium phosphate Substances 0.000 description 2
- 229910000389 calcium phosphate Inorganic materials 0.000 description 2
- 235000011010 calcium phosphates Nutrition 0.000 description 2
- 150000001721 carbon Chemical group 0.000 description 2
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
- 150000001732 carboxylic acid derivatives Chemical group 0.000 description 2
- 150000001733 carboxylic acid esters Chemical group 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 125000002091 cationic group Chemical group 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
- 229940106189 ceramide Drugs 0.000 description 2
- ZVEQCJWYRWKARO-UHFFFAOYSA-N ceramide Natural products CCCCCCCCCCCCCCC(O)C(=O)NC(CO)C(O)C=CCCC=C(C)CCCCCCCCC ZVEQCJWYRWKARO-UHFFFAOYSA-N 0.000 description 2
- 125000003636 chemical group Chemical group 0.000 description 2
- 230000003098 cholesteric effect Effects 0.000 description 2
- 238000004040 coloring Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 239000006071 cream Substances 0.000 description 2
- PCDQPRRSZKQHHS-ZAKLUEHWSA-N cytidine-5'-triphosphate Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@@H](O)[C@H](CO[P@](O)(=O)O[P@@](O)(=O)OP(O)(O)=O)O1 PCDQPRRSZKQHHS-ZAKLUEHWSA-N 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- ZCKYOWGFRHAZIQ-UHFFFAOYSA-N dihydrourocanic acid Chemical compound OC(=O)CCC1=CNC=N1 ZCKYOWGFRHAZIQ-UHFFFAOYSA-N 0.000 description 2
- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical class CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 description 2
- 238000002296 dynamic light scattering Methods 0.000 description 2
- 235000014103 egg white Nutrition 0.000 description 2
- 210000000969 egg white Anatomy 0.000 description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 2
- 102000013165 exonuclease Human genes 0.000 description 2
- 229960000961 floxuridine Drugs 0.000 description 2
- 125000003827 glycol group Chemical group 0.000 description 2
- 108010010147 glycylglutamine Proteins 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- 230000002440 hepatic effect Effects 0.000 description 2
- 125000005842 heteroatom Chemical group 0.000 description 2
- 150000002402 hexoses Chemical class 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 125000002883 imidazolyl group Chemical group 0.000 description 2
- 238000002991 immunohistochemical analysis Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 238000007918 intramuscular administration Methods 0.000 description 2
- 239000011630 iodine Substances 0.000 description 2
- 150000002576 ketones Chemical group 0.000 description 2
- 108010034529 leucyl-lysine Proteins 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 210000005229 liver cell Anatomy 0.000 description 2
- 230000033001 locomotion Effects 0.000 description 2
- 239000006210 lotion Substances 0.000 description 2
- 108010017391 lysylvaline Proteins 0.000 description 2
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 2
- 108010056582 methionylglutamic acid Proteins 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- YACKEPLHDIMKIO-UHFFFAOYSA-L methylphosphonate(2-) Chemical compound CP([O-])([O-])=O YACKEPLHDIMKIO-UHFFFAOYSA-L 0.000 description 2
- 238000001471 micro-filtration Methods 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- VVGIYYKRAMHVLU-UHFFFAOYSA-N newbouldiamide Natural products CCCCCCCCCCCCCCCCCCCC(O)C(O)C(O)C(CO)NC(=O)CCCCCCCCCCCCCCCCC VVGIYYKRAMHVLU-UHFFFAOYSA-N 0.000 description 2
- 230000037434 nonsense mutation Effects 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- GJVFBWCTGUSGDD-UHFFFAOYSA-L pentamethonium bromide Chemical compound [Br-].[Br-].C[N+](C)(C)CCCCC[N+](C)(C)C GJVFBWCTGUSGDD-UHFFFAOYSA-L 0.000 description 2
- 108010012581 phenylalanylglutamate Proteins 0.000 description 2
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 2
- 108010051242 phenylalanylserine Proteins 0.000 description 2
- 150000003904 phospholipids Chemical class 0.000 description 2
- PTMHPRAIXMAOOB-UHFFFAOYSA-L phosphoramidate Chemical class NP([O-])([O-])=O PTMHPRAIXMAOOB-UHFFFAOYSA-L 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 235000002378 plant sterols Nutrition 0.000 description 2
- 230000008488 polyadenylation Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 230000004481 post-translational protein modification Effects 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 108010029020 prolylglycine Proteins 0.000 description 2
- 235000013849 propane Nutrition 0.000 description 2
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- PTJWIQPHWPFNBW-GBNDHIKLSA-N pseudouridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1C1=CNC(=O)NC1=O PTJWIQPHWPFNBW-GBNDHIKLSA-N 0.000 description 2
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 2
- 150000003233 pyrroles Chemical group 0.000 description 2
- 239000010499 rapseed oil Substances 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 238000003259 recombinant expression Methods 0.000 description 2
- 210000000664 rectum Anatomy 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 2
- 239000004055 small Interfering RNA Substances 0.000 description 2
- 108091029842 small nuclear ribonucleic acid Proteins 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 230000006641 stabilisation Effects 0.000 description 2
- 238000011105 stabilization Methods 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 2
- 229910052717 sulfur Inorganic materials 0.000 description 2
- 210000000106 sweat gland Anatomy 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 238000009210 therapy by ultrasound Methods 0.000 description 2
- 229940104230 thymidine Drugs 0.000 description 2
- 210000003437 trachea Anatomy 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 2
- DCXXMTOCNZCJGO-UHFFFAOYSA-N tristearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC DCXXMTOCNZCJGO-UHFFFAOYSA-N 0.000 description 2
- 108010051110 tyrosyl-lysine Proteins 0.000 description 2
- DRTQHJPVMGBUCF-UHFFFAOYSA-N uracil arabinoside Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-UHFFFAOYSA-N 0.000 description 2
- PGAVKCOVUIYSFO-UHFFFAOYSA-N uridine-triphosphate Natural products OC1C(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)OC1N1C(=O)NC(=O)C=C1 PGAVKCOVUIYSFO-UHFFFAOYSA-N 0.000 description 2
- IXOXBSCIXZEQEQ-KQYNXXCUSA-N (2r,3r,4s,5r)-2-(2-amino-6-methoxypurin-9-yl)-5-(hydroxymethyl)oxolane-3,4-diol Chemical compound C1=NC=2C(OC)=NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O IXOXBSCIXZEQEQ-KQYNXXCUSA-N 0.000 description 1
- OPCHFPHZPIURNA-MFERNQICSA-N (2s)-2,5-bis(3-aminopropylamino)-n-[2-(dioctadecylamino)acetyl]pentanamide Chemical compound CCCCCCCCCCCCCCCCCCN(CC(=O)NC(=O)[C@H](CCCNCCCN)NCCCN)CCCCCCCCCCCCCCCCCC OPCHFPHZPIURNA-MFERNQICSA-N 0.000 description 1
- DSSYKIVIOFKYAU-XCBNKYQSSA-N (R)-camphor Chemical compound C1C[C@@]2(C)C(=O)C[C@@H]1C2(C)C DSSYKIVIOFKYAU-XCBNKYQSSA-N 0.000 description 1
- IUPSCXDOKZWYRB-UHFFFAOYSA-N 1,2,3$l^{2}-triphosphirene Chemical compound [P]1P=P1 IUPSCXDOKZWYRB-UHFFFAOYSA-N 0.000 description 1
- NBKORJKMMVZAOZ-VPCXQMTMSA-N 1-[(2r,3r,4r,5r)-3,4-dihydroxy-5-(hydroxymethyl)-3-methyloxolan-2-yl]pyrimidine-2,4-dione Chemical compound C[C@@]1(O)[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 NBKORJKMMVZAOZ-VPCXQMTMSA-N 0.000 description 1
- SATCOUWSAZBIJO-UHFFFAOYSA-N 1-methyladenine Natural products N=C1N(C)C=NC2=C1NC=N2 SATCOUWSAZBIJO-UHFFFAOYSA-N 0.000 description 1
- GFYLSDSUCHVORB-IOSLPCCCSA-N 1-methyladenosine Chemical compound C1=NC=2C(=N)N(C)C=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O GFYLSDSUCHVORB-IOSLPCCCSA-N 0.000 description 1
- UTAIYTHAJQNQDW-KQYNXXCUSA-N 1-methylguanosine Chemical compound C1=NC=2C(=O)N(C)C(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O UTAIYTHAJQNQDW-KQYNXXCUSA-N 0.000 description 1
- WJNGQIYEQLPJMN-IOSLPCCCSA-N 1-methylinosine Chemical compound C1=NC=2C(=O)N(C)C=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O WJNGQIYEQLPJMN-IOSLPCCCSA-N 0.000 description 1
- YKBGVTZYEHREMT-KVQBGUIXSA-N 2'-deoxyguanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](CO)O1 YKBGVTZYEHREMT-KVQBGUIXSA-N 0.000 description 1
- CKTSBUTUHBMZGZ-SHYZEUOFSA-N 2'‐deoxycytidine Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 CKTSBUTUHBMZGZ-SHYZEUOFSA-N 0.000 description 1
- ZENKESXKWBIZCV-UHFFFAOYSA-N 2,2,4,4-tetrafluoro-1,3-benzodioxin-6-amine Chemical group O1C(F)(F)OC(F)(F)C2=CC(N)=CC=C21 ZENKESXKWBIZCV-UHFFFAOYSA-N 0.000 description 1
- HLYBTPMYFWWNJN-UHFFFAOYSA-N 2-(2,4-dioxo-1h-pyrimidin-5-yl)-2-hydroxyacetic acid Chemical compound OC(=O)C(O)C1=CNC(=O)NC1=O HLYBTPMYFWWNJN-UHFFFAOYSA-N 0.000 description 1
- HCYQTNUNMRYQLZ-UHFFFAOYSA-N 2-[(6-methyl-2,4-dioxo-1H-pyrimidin-5-yl)amino]acetic acid Chemical compound C(=O)(O)CNC=1C(NC(NC=1C)=O)=O HCYQTNUNMRYQLZ-UHFFFAOYSA-N 0.000 description 1
- PGYFLJKHWJVRMC-ZXRZDOCRSA-N 2-[4-[[(3s,8s,9s,10r,13r,14s,17r)-10,13-dimethyl-17-[(2r)-6-methylheptan-2-yl]-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthren-3-yl]oxy]butoxy]-n,n-dimethyl-3-[(9z,12z)-octadeca-9,12-dienoxy]propan-1-amine Chemical compound C([C@@H]12)C[C@]3(C)[C@@H]([C@H](C)CCCC(C)C)CC[C@H]3[C@@H]1CC=C1[C@]2(C)CC[C@H](OCCCCOC(CN(C)C)COCCCCCCCC\C=C/C\C=C/CCCCC)C1 PGYFLJKHWJVRMC-ZXRZDOCRSA-N 0.000 description 1
- DQVAZKGVGKHQDS-UHFFFAOYSA-N 2-[[1-[2-[(2-amino-4-methylpentanoyl)amino]-4-methylpentanoyl]pyrrolidine-2-carbonyl]amino]-4-methylpentanoic acid Chemical compound CC(C)CC(N)C(=O)NC(CC(C)C)C(=O)N1CCCC1C(=O)NC(CC(C)C)C(O)=O DQVAZKGVGKHQDS-UHFFFAOYSA-N 0.000 description 1
- CZINFFCCOSHTMZ-NYVOMTAGSA-N 2-[bis[(Z)-octadec-9-enoyl]amino]ethyl [(2R)-2,3-dihydroxypropyl] hydrogen phosphate Chemical compound C(CCCCCCC\C=C/CCCCCCCC)(=O)N(CCOP(OC[C@@H](CO)O)(=O)O)C(CCCCCCC\C=C/CCCCCCCC)=O CZINFFCCOSHTMZ-NYVOMTAGSA-N 0.000 description 1
- MWBWWFOAEOYUST-UHFFFAOYSA-N 2-aminopurine Chemical compound NC1=NC=C2N=CNC2=N1 MWBWWFOAEOYUST-UHFFFAOYSA-N 0.000 description 1
- HBJGQJWNMZDFKL-UHFFFAOYSA-N 2-chloro-7h-purin-6-amine Chemical compound NC1=NC(Cl)=NC2=C1NC=N2 HBJGQJWNMZDFKL-UHFFFAOYSA-N 0.000 description 1
- SMADWRYCYBUIKH-UHFFFAOYSA-N 2-methyl-7h-purin-6-amine Chemical compound CC1=NC(N)=C2NC=NC2=N1 SMADWRYCYBUIKH-UHFFFAOYSA-N 0.000 description 1
- VJROPLWGFCORRM-UHFFFAOYSA-N 2-methylbutan-1-amine Chemical compound CCC(C)CN VJROPLWGFCORRM-UHFFFAOYSA-N 0.000 description 1
- 229940080296 2-naphthalenesulfonate Drugs 0.000 description 1
- GJTBSTBJLVYKAU-XVFCMESISA-N 2-thiouridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=S)NC(=O)C=C1 GJTBSTBJLVYKAU-XVFCMESISA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- ALKYHXVLJMQRLQ-UHFFFAOYSA-M 3-carboxynaphthalen-2-olate Chemical compound C1=CC=C2C=C(C([O-])=O)C(O)=CC2=C1 ALKYHXVLJMQRLQ-UHFFFAOYSA-M 0.000 description 1
- 125000004080 3-carboxypropanoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C(O[H])=O 0.000 description 1
- KOLPWZCZXAMXKS-UHFFFAOYSA-N 3-methylcytosine Chemical compound CN1C(N)=CC=NC1=O KOLPWZCZXAMXKS-UHFFFAOYSA-N 0.000 description 1
- XMIIGOLPHOKFCH-UHFFFAOYSA-N 3-phenylpropionic acid Chemical class OC(=O)CCC1=CC=CC=C1 XMIIGOLPHOKFCH-UHFFFAOYSA-N 0.000 description 1
- FEOYKPQEERVCAV-XVFCMESISA-N 4-amino-1-[(2r,3r,4s,5r)-3-azido-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-2-one Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](N=[N+]=[N-])[C@H](O)[C@@H](CO)O1 FEOYKPQEERVCAV-XVFCMESISA-N 0.000 description 1
- LNRZWAHOOLIOLA-MYINAIGISA-N 4-amino-1-[(2s,4s,5r)-2-fluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-2-one Chemical compound O=C1N=C(N)C=CN1[C@]1(F)O[C@H](CO)[C@@H](O)C1 LNRZWAHOOLIOLA-MYINAIGISA-N 0.000 description 1
- MQJSSLBGAQJNER-UHFFFAOYSA-N 5-(methylaminomethyl)-1h-pyrimidine-2,4-dione Chemical compound CNCC1=CNC(=O)NC1=O MQJSSLBGAQJNER-UHFFFAOYSA-N 0.000 description 1
- OZQDLJNDRVBCST-SHUUEZRQSA-N 5-amino-2-[(2r,3r,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-1,2,4-triazin-3-one Chemical compound O=C1N=C(N)C=NN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 OZQDLJNDRVBCST-SHUUEZRQSA-N 0.000 description 1
- UJBCLAXPPIDQEE-UHFFFAOYSA-N 5-prop-1-ynyl-1h-pyrimidine-2,4-dione Chemical compound CC#CC1=CNC(=O)NC1=O UJBCLAXPPIDQEE-UHFFFAOYSA-N 0.000 description 1
- DCPSTSVLRXOYGS-UHFFFAOYSA-N 6-amino-1h-pyrimidine-2-thione Chemical compound NC1=CC=NC(S)=N1 DCPSTSVLRXOYGS-UHFFFAOYSA-N 0.000 description 1
- CKOMXBHMKXXTNW-UHFFFAOYSA-N 6-methyladenine Chemical compound CNC1=NC=NC2=C1N=CN2 CKOMXBHMKXXTNW-UHFFFAOYSA-N 0.000 description 1
- OGHAROSJZRTIOK-KQYNXXCUSA-O 7-methylguanosine Chemical compound C1=2N=C(N)NC(=O)C=2[N+](C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OGHAROSJZRTIOK-KQYNXXCUSA-O 0.000 description 1
- YMVDTXSRLFAIKI-UHFFFAOYSA-N 7h-purine Chemical compound C1=NC=C2NC=NC2=N1.C1=NC=C2NC=NC2=N1 YMVDTXSRLFAIKI-UHFFFAOYSA-N 0.000 description 1
- VKKXEIQIGGPMHT-UHFFFAOYSA-N 7h-purine-2,8-diamine Chemical compound NC1=NC=C2NC(N)=NC2=N1 VKKXEIQIGGPMHT-UHFFFAOYSA-N 0.000 description 1
- MSSXOMSJDRHRMC-UHFFFAOYSA-N 9H-purine-2,6-diamine Chemical compound NC1=NC(N)=C2NC=NC2=N1 MSSXOMSJDRHRMC-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- ZKHQWZAMYRWXGA-KQYNXXCUSA-N Adenosine triphosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-N 0.000 description 1
- SKHCUBQVZJHOFM-NAKRPEOUSA-N Ala-Arg-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O SKHCUBQVZJHOFM-NAKRPEOUSA-N 0.000 description 1
- TTXMOJWKNRJWQJ-FXQIFTODSA-N Ala-Arg-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CCCN=C(N)N TTXMOJWKNRJWQJ-FXQIFTODSA-N 0.000 description 1
- XQGIRPGAVLFKBJ-CIUDSAMLSA-N Ala-Asn-Lys Chemical compound N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)O XQGIRPGAVLFKBJ-CIUDSAMLSA-N 0.000 description 1
- MBWYUTNBYSSUIQ-HERUPUMHSA-N Ala-Asn-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N MBWYUTNBYSSUIQ-HERUPUMHSA-N 0.000 description 1
- WDIYWDJLXOCGRW-ACZMJKKPSA-N Ala-Asp-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O WDIYWDJLXOCGRW-ACZMJKKPSA-N 0.000 description 1
- BLIMFWGRQKRCGT-YUMQZZPRSA-N Ala-Gly-Lys Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN BLIMFWGRQKRCGT-YUMQZZPRSA-N 0.000 description 1
- TZDNWXDLYFIFPT-BJDJZHNGSA-N Ala-Ile-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O TZDNWXDLYFIFPT-BJDJZHNGSA-N 0.000 description 1
- HHRAXZAYZFFRAM-CIUDSAMLSA-N Ala-Leu-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O HHRAXZAYZFFRAM-CIUDSAMLSA-N 0.000 description 1
- AWZKCUCQJNTBAD-SRVKXCTJSA-N Ala-Leu-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN AWZKCUCQJNTBAD-SRVKXCTJSA-N 0.000 description 1
- RAAWHFXHAACDFT-FXQIFTODSA-N Ala-Met-Asn Chemical compound CSCC[C@H](NC(=O)[C@H](C)N)C(=O)N[C@@H](CC(N)=O)C(O)=O RAAWHFXHAACDFT-FXQIFTODSA-N 0.000 description 1
- DHBKYZYFEXXUAK-ONGXEEELSA-N Ala-Phe-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 DHBKYZYFEXXUAK-ONGXEEELSA-N 0.000 description 1
- FEGOCLZUJUFCHP-CIUDSAMLSA-N Ala-Pro-Gln Chemical compound [H]N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O FEGOCLZUJUFCHP-CIUDSAMLSA-N 0.000 description 1
- SYIFFFHSXBNPMC-UWJYBYFXSA-N Ala-Ser-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N SYIFFFHSXBNPMC-UWJYBYFXSA-N 0.000 description 1
- LYILPUNCKACNGF-NAKRPEOUSA-N Ala-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C)N LYILPUNCKACNGF-NAKRPEOUSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 241000272525 Anas platyrhynchos Species 0.000 description 1
- 108091023037 Aptamer Proteins 0.000 description 1
- VWVPYNGMOCSSGK-GUBZILKMSA-N Arg-Arg-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O VWVPYNGMOCSSGK-GUBZILKMSA-N 0.000 description 1
- KJGNDQCYBNBXDA-GUBZILKMSA-N Arg-Arg-Cys Chemical compound C(C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CS)C(=O)O)N)CN=C(N)N KJGNDQCYBNBXDA-GUBZILKMSA-N 0.000 description 1
- MUXONAMCEUBVGA-DCAQKATOSA-N Arg-Arg-Gln Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(N)=O)C(O)=O MUXONAMCEUBVGA-DCAQKATOSA-N 0.000 description 1
- IIABBYGHLYWVOS-FXQIFTODSA-N Arg-Asn-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O IIABBYGHLYWVOS-FXQIFTODSA-N 0.000 description 1
- FEZJJKXNPSEYEV-CIUDSAMLSA-N Arg-Gln-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O FEZJJKXNPSEYEV-CIUDSAMLSA-N 0.000 description 1
- YHQGEARSFILVHL-HJGDQZAQSA-N Arg-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCN=C(N)N)N)O YHQGEARSFILVHL-HJGDQZAQSA-N 0.000 description 1
- JQFJNGVSGOUQDH-XIRDDKMYSA-N Arg-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCN=C(N)N)N)C(O)=O)=CNC2=C1 JQFJNGVSGOUQDH-XIRDDKMYSA-N 0.000 description 1
- AGVNTAUPLWIQEN-ZPFDUUQYSA-N Arg-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N AGVNTAUPLWIQEN-ZPFDUUQYSA-N 0.000 description 1
- FFEUXEAKYRCACT-PEDHHIEDSA-N Arg-Ile-Ile Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCCNC(N)=N)[C@@H](C)CC)C(O)=O FFEUXEAKYRCACT-PEDHHIEDSA-N 0.000 description 1
- GXXWTNKNFFKTJB-NAKRPEOUSA-N Arg-Ile-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O GXXWTNKNFFKTJB-NAKRPEOUSA-N 0.000 description 1
- COXMUHNBYCVVRG-DCAQKATOSA-N Arg-Leu-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O COXMUHNBYCVVRG-DCAQKATOSA-N 0.000 description 1
- RIQBRKVTFBWEDY-RHYQMDGZSA-N Arg-Lys-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O RIQBRKVTFBWEDY-RHYQMDGZSA-N 0.000 description 1
- ICRHGPYYXMWHIE-LPEHRKFASA-N Arg-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O ICRHGPYYXMWHIE-LPEHRKFASA-N 0.000 description 1
- QMQZYILAWUOLPV-JYJNAYRXSA-N Arg-Tyr-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)CC1=CC=C(O)C=C1 QMQZYILAWUOLPV-JYJNAYRXSA-N 0.000 description 1
- XYOVHPDDWCEUDY-CIUDSAMLSA-N Asn-Ala-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O XYOVHPDDWCEUDY-CIUDSAMLSA-N 0.000 description 1
- HAJWYALLJIATCX-FXQIFTODSA-N Asn-Asn-Arg Chemical compound C(C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)N)N)CN=C(N)N HAJWYALLJIATCX-FXQIFTODSA-N 0.000 description 1
- DAPLJWATMAXPPZ-CIUDSAMLSA-N Asn-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(N)=O DAPLJWATMAXPPZ-CIUDSAMLSA-N 0.000 description 1
- KXFCBAHYSLJCCY-ZLUOBGJFSA-N Asn-Asn-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O KXFCBAHYSLJCCY-ZLUOBGJFSA-N 0.000 description 1
- VJTWLBMESLDOMK-WDSKDSINSA-N Asn-Gln-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O VJTWLBMESLDOMK-WDSKDSINSA-N 0.000 description 1
- PPMTUXJSQDNUDE-CIUDSAMLSA-N Asn-Glu-Arg Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N PPMTUXJSQDNUDE-CIUDSAMLSA-N 0.000 description 1
- IICZCLFBILYRCU-WHFBIAKZSA-N Asn-Gly-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IICZCLFBILYRCU-WHFBIAKZSA-N 0.000 description 1
- SPCONPVIDFMDJI-QSFUFRPTSA-N Asn-Ile-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O SPCONPVIDFMDJI-QSFUFRPTSA-N 0.000 description 1
- BXUHCIXDSWRSBS-CIUDSAMLSA-N Asn-Leu-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O BXUHCIXDSWRSBS-CIUDSAMLSA-N 0.000 description 1
- FHETWELNCBMRMG-HJGDQZAQSA-N Asn-Leu-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FHETWELNCBMRMG-HJGDQZAQSA-N 0.000 description 1
- FBODFHMLALOPHP-GUBZILKMSA-N Asn-Lys-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O FBODFHMLALOPHP-GUBZILKMSA-N 0.000 description 1
- JWKDQOORUCYUIW-ZPFDUUQYSA-N Asn-Lys-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JWKDQOORUCYUIW-ZPFDUUQYSA-N 0.000 description 1
- WXVGISRWSYGEDK-KKUMJFAQSA-N Asn-Lys-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(=O)N)N WXVGISRWSYGEDK-KKUMJFAQSA-N 0.000 description 1
- RVHGJNGNKGDCPX-KKUMJFAQSA-N Asn-Phe-Lys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N RVHGJNGNKGDCPX-KKUMJFAQSA-N 0.000 description 1
- DOURAOODTFJRIC-CIUDSAMLSA-N Asn-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)N DOURAOODTFJRIC-CIUDSAMLSA-N 0.000 description 1
- WSOKZUVWBXVJHX-CIUDSAMLSA-N Asp-Arg-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O WSOKZUVWBXVJHX-CIUDSAMLSA-N 0.000 description 1
- TVVYVAUGRHNTGT-UGYAYLCHSA-N Asp-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(O)=O TVVYVAUGRHNTGT-UGYAYLCHSA-N 0.000 description 1
- RYKWOUUZJFSJOH-FXQIFTODSA-N Asp-Gln-Glu Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)O)N RYKWOUUZJFSJOH-FXQIFTODSA-N 0.000 description 1
- RRKCPMGSRIDLNC-AVGNSLFASA-N Asp-Glu-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O RRKCPMGSRIDLNC-AVGNSLFASA-N 0.000 description 1
- WSGVTKZFVJSJOG-RCOVLWMOSA-N Asp-Gly-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O WSGVTKZFVJSJOG-RCOVLWMOSA-N 0.000 description 1
- KLYPOCBLKMPBIQ-GHCJXIJMSA-N Asp-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC(=O)O)N KLYPOCBLKMPBIQ-GHCJXIJMSA-N 0.000 description 1
- RTXQQDVBACBSCW-CFMVVWHZSA-N Asp-Ile-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O RTXQQDVBACBSCW-CFMVVWHZSA-N 0.000 description 1
- JXGJJQJHXHXJQF-CIUDSAMLSA-N Asp-Met-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(O)=O JXGJJQJHXHXJQF-CIUDSAMLSA-N 0.000 description 1
- GPPIDDWYKJPRES-YDHLFZDLSA-N Asp-Phe-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O GPPIDDWYKJPRES-YDHLFZDLSA-N 0.000 description 1
- KESWRFKUZRUTAH-FXQIFTODSA-N Asp-Pro-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O KESWRFKUZRUTAH-FXQIFTODSA-N 0.000 description 1
- CUQDCPXNZPDYFQ-ZLUOBGJFSA-N Asp-Ser-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O CUQDCPXNZPDYFQ-ZLUOBGJFSA-N 0.000 description 1
- QSFHZPQUAAQHAQ-CIUDSAMLSA-N Asp-Ser-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O QSFHZPQUAAQHAQ-CIUDSAMLSA-N 0.000 description 1
- MJJIHRWNWSQTOI-VEVYYDQMSA-N Asp-Thr-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O MJJIHRWNWSQTOI-VEVYYDQMSA-N 0.000 description 1
- 241000972773 Aulopiformes Species 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 description 1
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Natural products CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 1
- MKYMAGPSMUAMQP-ADGNVCHCSA-N C(C)O.P(=O)(O)(O)OC([C@@H](C(O)C(CCCCCCCCCCCCC)=O)O)C(CCCCCCCCCCCCC)=O Chemical compound C(C)O.P(=O)(O)(O)OC([C@@H](C(O)C(CCCCCCCCCCCCC)=O)O)C(CCCCCCCCCCCCC)=O MKYMAGPSMUAMQP-ADGNVCHCSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000282836 Camelus dromedarius Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 108090000565 Capsid Proteins Proteins 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 102100023321 Ceruloplasmin Human genes 0.000 description 1
- 241000723346 Cinnamomum camphora Species 0.000 description 1
- 108020004638 Circular DNA Proteins 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- YZFCGHIBLBDZDA-ZLUOBGJFSA-N Cys-Asp-Ser Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O YZFCGHIBLBDZDA-ZLUOBGJFSA-N 0.000 description 1
- PDRMRVHPAQKTLT-NAKRPEOUSA-N Cys-Ile-Val Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O PDRMRVHPAQKTLT-NAKRPEOUSA-N 0.000 description 1
- WKKKNGNJDGATNS-QEJZJMRPSA-N Cys-Trp-Glu Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(O)=O WKKKNGNJDGATNS-QEJZJMRPSA-N 0.000 description 1
- ODKSFYDXXFIFQN-SCSAIBSYSA-N D-arginine Chemical compound OC(=O)[C@H](N)CCCNC(N)=N ODKSFYDXXFIFQN-SCSAIBSYSA-N 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 description 1
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 1
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 1
- CKTSBUTUHBMZGZ-UHFFFAOYSA-N Deoxycytidine Natural products O=C1N=C(N)C=CN1C1OC(CO)C(O)C1 CKTSBUTUHBMZGZ-UHFFFAOYSA-N 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical class SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 description 1
- SNRUBQQJIBEYMU-UHFFFAOYSA-N Dodecane Natural products CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 241000699694 Gerbillinae Species 0.000 description 1
- INKFLNZBTSNFON-CIUDSAMLSA-N Gln-Ala-Arg Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O INKFLNZBTSNFON-CIUDSAMLSA-N 0.000 description 1
- RGXXLQWXBFNXTG-CIUDSAMLSA-N Gln-Arg-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O RGXXLQWXBFNXTG-CIUDSAMLSA-N 0.000 description 1
- PRBLYKYHAJEABA-SRVKXCTJSA-N Gln-Arg-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O PRBLYKYHAJEABA-SRVKXCTJSA-N 0.000 description 1
- WMOMPXKOKASNBK-PEFMBERDSA-N Gln-Asn-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WMOMPXKOKASNBK-PEFMBERDSA-N 0.000 description 1
- AAOBFSKXAVIORT-GUBZILKMSA-N Gln-Asn-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O AAOBFSKXAVIORT-GUBZILKMSA-N 0.000 description 1
- KZEUVLLVULIPNX-GUBZILKMSA-N Gln-Asp-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)N)N KZEUVLLVULIPNX-GUBZILKMSA-N 0.000 description 1
- JKPGHIQCHIIRMS-AVGNSLFASA-N Gln-Asp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)N)N JKPGHIQCHIIRMS-AVGNSLFASA-N 0.000 description 1
- LVNILKSSFHCSJZ-IHRRRGAJSA-N Gln-Gln-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)N)N LVNILKSSFHCSJZ-IHRRRGAJSA-N 0.000 description 1
- DRDSQGHKTLSNEA-GLLZPBPUSA-N Gln-Glu-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O DRDSQGHKTLSNEA-GLLZPBPUSA-N 0.000 description 1
- JHPFPROFOAJRFN-IHRRRGAJSA-N Gln-Glu-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCC(=O)N)N)O JHPFPROFOAJRFN-IHRRRGAJSA-N 0.000 description 1
- KKCJHBXMYYVWMX-KQXIARHKSA-N Gln-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N KKCJHBXMYYVWMX-KQXIARHKSA-N 0.000 description 1
- XFAUJGNLHIGXET-AVGNSLFASA-N Gln-Leu-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O XFAUJGNLHIGXET-AVGNSLFASA-N 0.000 description 1
- IULKWYSYZSURJK-AVGNSLFASA-N Gln-Leu-Lys Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O IULKWYSYZSURJK-AVGNSLFASA-N 0.000 description 1
- YPMDZWPZFOZYFG-GUBZILKMSA-N Gln-Leu-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YPMDZWPZFOZYFG-GUBZILKMSA-N 0.000 description 1
- QKWBEMCLYTYBNI-GVXVVHGQSA-N Gln-Lys-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCC(N)=O QKWBEMCLYTYBNI-GVXVVHGQSA-N 0.000 description 1
- LUGUNEGJNDEBLU-DCAQKATOSA-N Gln-Met-Arg Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N LUGUNEGJNDEBLU-DCAQKATOSA-N 0.000 description 1
- UESYBOXFJWJVSB-AVGNSLFASA-N Gln-Phe-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O UESYBOXFJWJVSB-AVGNSLFASA-N 0.000 description 1
- DOQUICBEISTQHE-CIUDSAMLSA-N Gln-Pro-Asp Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O DOQUICBEISTQHE-CIUDSAMLSA-N 0.000 description 1
- ZGHMRONFHDVXEF-AVGNSLFASA-N Gln-Ser-Phe Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ZGHMRONFHDVXEF-AVGNSLFASA-N 0.000 description 1
- ININBLZFFVOQIO-JHEQGTHGSA-N Gln-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCC(=O)N)N)O ININBLZFFVOQIO-JHEQGTHGSA-N 0.000 description 1
- XMWNHGKDDIFXQJ-NWLDYVSISA-N Gln-Thr-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O XMWNHGKDDIFXQJ-NWLDYVSISA-N 0.000 description 1
- RBSKVTZUFMIWFU-XEGUGMAKSA-N Gln-Trp-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C)C(O)=O RBSKVTZUFMIWFU-XEGUGMAKSA-N 0.000 description 1
- CVRUVYDNRPSKBM-QEJZJMRPSA-N Gln-Trp-Ser Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)N)N CVRUVYDNRPSKBM-QEJZJMRPSA-N 0.000 description 1
- RLZBLVSJDFHDBL-KBIXCLLPSA-N Glu-Ala-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O RLZBLVSJDFHDBL-KBIXCLLPSA-N 0.000 description 1
- WOMUDRVDJMHTCV-DCAQKATOSA-N Glu-Arg-Arg Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WOMUDRVDJMHTCV-DCAQKATOSA-N 0.000 description 1
- NLKVNZUFDPWPNL-YUMQZZPRSA-N Glu-Arg-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O NLKVNZUFDPWPNL-YUMQZZPRSA-N 0.000 description 1
- KKCUFHUTMKQQCF-SRVKXCTJSA-N Glu-Arg-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O KKCUFHUTMKQQCF-SRVKXCTJSA-N 0.000 description 1
- WOSRKEJQESVHGA-CIUDSAMLSA-N Glu-Arg-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O WOSRKEJQESVHGA-CIUDSAMLSA-N 0.000 description 1
- AFODTOLGSZQDSL-PEFMBERDSA-N Glu-Asn-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCC(=O)O)N AFODTOLGSZQDSL-PEFMBERDSA-N 0.000 description 1
- RJONUNZIMUXUOI-GUBZILKMSA-N Glu-Asn-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCC(=O)O)N RJONUNZIMUXUOI-GUBZILKMSA-N 0.000 description 1
- PXHABOCPJVTGEK-BQBZGAKWSA-N Glu-Gln-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O PXHABOCPJVTGEK-BQBZGAKWSA-N 0.000 description 1
- NKLRYVLERDYDBI-FXQIFTODSA-N Glu-Glu-Asp Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O NKLRYVLERDYDBI-FXQIFTODSA-N 0.000 description 1
- PHONAZGUEGIOEM-GLLZPBPUSA-N Glu-Glu-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PHONAZGUEGIOEM-GLLZPBPUSA-N 0.000 description 1
- PXXGVUVQWQGGIG-YUMQZZPRSA-N Glu-Gly-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N PXXGVUVQWQGGIG-YUMQZZPRSA-N 0.000 description 1
- OGNJZUXUTPQVBR-BQBZGAKWSA-N Glu-Gly-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O OGNJZUXUTPQVBR-BQBZGAKWSA-N 0.000 description 1
- RAUDKMVXNOWDLS-WDSKDSINSA-N Glu-Gly-Ser Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O RAUDKMVXNOWDLS-WDSKDSINSA-N 0.000 description 1
- CXRWMMRLEMVSEH-PEFMBERDSA-N Glu-Ile-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O CXRWMMRLEMVSEH-PEFMBERDSA-N 0.000 description 1
- PJBVXVBTTFZPHJ-GUBZILKMSA-N Glu-Leu-Asp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)O)N PJBVXVBTTFZPHJ-GUBZILKMSA-N 0.000 description 1
- IRXNJYPKBVERCW-DCAQKATOSA-N Glu-Leu-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IRXNJYPKBVERCW-DCAQKATOSA-N 0.000 description 1
- IVGJYOOGJLFKQE-AVGNSLFASA-N Glu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N IVGJYOOGJLFKQE-AVGNSLFASA-N 0.000 description 1
- WNRZUESNGGDCJX-JYJNAYRXSA-N Glu-Leu-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O WNRZUESNGGDCJX-JYJNAYRXSA-N 0.000 description 1
- FBEJIDRSQCGFJI-GUBZILKMSA-N Glu-Leu-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O FBEJIDRSQCGFJI-GUBZILKMSA-N 0.000 description 1
- SJJHXJDSNQJMMW-SRVKXCTJSA-N Glu-Lys-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O SJJHXJDSNQJMMW-SRVKXCTJSA-N 0.000 description 1
- CUPSDFQZTVVTSK-GUBZILKMSA-N Glu-Lys-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCC(O)=O CUPSDFQZTVVTSK-GUBZILKMSA-N 0.000 description 1
- BCYGDJXHAGZNPQ-DCAQKATOSA-N Glu-Lys-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O BCYGDJXHAGZNPQ-DCAQKATOSA-N 0.000 description 1
- ZGEJRLJEAMPEDV-SRVKXCTJSA-N Glu-Lys-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(=O)O)N ZGEJRLJEAMPEDV-SRVKXCTJSA-N 0.000 description 1
- YRMZCZIRHYCNHX-RYUDHWBXSA-N Glu-Phe-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(O)=O YRMZCZIRHYCNHX-RYUDHWBXSA-N 0.000 description 1
- SYWCGQOIIARSIX-SRVKXCTJSA-N Glu-Pro-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O SYWCGQOIIARSIX-SRVKXCTJSA-N 0.000 description 1
- YQAQQKPWFOBSMU-WDCWCFNPSA-N Glu-Thr-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O YQAQQKPWFOBSMU-WDCWCFNPSA-N 0.000 description 1
- YQPFCZVKMUVZIN-AUTRQRHGSA-N Glu-Val-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O YQPFCZVKMUVZIN-AUTRQRHGSA-N 0.000 description 1
- SOYWRINXUSUWEQ-DLOVCJGASA-N Glu-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCC(O)=O SOYWRINXUSUWEQ-DLOVCJGASA-N 0.000 description 1
- RQZGFWKQLPJOEQ-YUMQZZPRSA-N Gly-Arg-Gln Chemical compound C(C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)CN)CN=C(N)N RQZGFWKQLPJOEQ-YUMQZZPRSA-N 0.000 description 1
- MXXXVOYFNVJHMA-IUCAKERBSA-N Gly-Arg-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CN MXXXVOYFNVJHMA-IUCAKERBSA-N 0.000 description 1
- XCLCVBYNGXEVDU-WHFBIAKZSA-N Gly-Asn-Ser Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O XCLCVBYNGXEVDU-WHFBIAKZSA-N 0.000 description 1
- KQDMENMTYNBWMR-WHFBIAKZSA-N Gly-Asp-Ala Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O KQDMENMTYNBWMR-WHFBIAKZSA-N 0.000 description 1
- PEZZSFLFXXFUQD-XPUUQOCRSA-N Gly-Cys-Val Chemical compound [H]NCC(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O PEZZSFLFXXFUQD-XPUUQOCRSA-N 0.000 description 1
- XTQFHTHIAKKCTM-YFKPBYRVSA-N Gly-Glu-Gly Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O XTQFHTHIAKKCTM-YFKPBYRVSA-N 0.000 description 1
- KMSGYZQRXPUKGI-BYPYZUCNSA-N Gly-Gly-Asn Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CC(N)=O KMSGYZQRXPUKGI-BYPYZUCNSA-N 0.000 description 1
- QPTNELDXWKRIFX-YFKPBYRVSA-N Gly-Gly-Gln Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCC(N)=O QPTNELDXWKRIFX-YFKPBYRVSA-N 0.000 description 1
- XMPXVJIDADUOQB-RCOVLWMOSA-N Gly-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C([O-])=O)NC(=O)CNC(=O)C[NH3+] XMPXVJIDADUOQB-RCOVLWMOSA-N 0.000 description 1
- MVORZMQFXBLMHM-QWRGUYRKSA-N Gly-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 MVORZMQFXBLMHM-QWRGUYRKSA-N 0.000 description 1
- SCWYHUQOOFRVHP-MBLNEYKQSA-N Gly-Ile-Thr Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SCWYHUQOOFRVHP-MBLNEYKQSA-N 0.000 description 1
- ULZCYBYDTUMHNF-IUCAKERBSA-N Gly-Leu-Glu Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O ULZCYBYDTUMHNF-IUCAKERBSA-N 0.000 description 1
- PTIIBFKSLCYQBO-NHCYSSNCSA-N Gly-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)CN PTIIBFKSLCYQBO-NHCYSSNCSA-N 0.000 description 1
- MHXKHKWHPNETGG-QWRGUYRKSA-N Gly-Lys-Leu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O MHXKHKWHPNETGG-QWRGUYRKSA-N 0.000 description 1
- NTBOEZICHOSJEE-YUMQZZPRSA-N Gly-Lys-Ser Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O NTBOEZICHOSJEE-YUMQZZPRSA-N 0.000 description 1
- IGOYNRWLWHWAQO-JTQLQIEISA-N Gly-Phe-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 IGOYNRWLWHWAQO-JTQLQIEISA-N 0.000 description 1
- FEUPVVCGQLNXNP-IRXDYDNUSA-N Gly-Phe-Phe Chemical compound C([C@H](NC(=O)CN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 FEUPVVCGQLNXNP-IRXDYDNUSA-N 0.000 description 1
- HUFUVTYGPOUCBN-MBLNEYKQSA-N Gly-Thr-Ile Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HUFUVTYGPOUCBN-MBLNEYKQSA-N 0.000 description 1
- LLWQVJNHMYBLLK-CDMKHQONSA-N Gly-Thr-Phe Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LLWQVJNHMYBLLK-CDMKHQONSA-N 0.000 description 1
- GWNIGUKSRJBIHX-STQMWFEESA-N Gly-Tyr-Arg Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)CN)O GWNIGUKSRJBIHX-STQMWFEESA-N 0.000 description 1
- SBVMXEZQJVUARN-XPUUQOCRSA-N Gly-Val-Ser Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O SBVMXEZQJVUARN-XPUUQOCRSA-N 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- JBJNKUOMNZGQIM-PYJNHQTQSA-N His-Arg-Ile Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JBJNKUOMNZGQIM-PYJNHQTQSA-N 0.000 description 1
- VFBZWZXKCVBTJR-SRVKXCTJSA-N His-Leu-Asp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N VFBZWZXKCVBTJR-SRVKXCTJSA-N 0.000 description 1
- RNMNYMDTESKEAJ-KKUMJFAQSA-N His-Leu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CN=CN1 RNMNYMDTESKEAJ-KKUMJFAQSA-N 0.000 description 1
- TWROVBNEHJSXDG-IHRRRGAJSA-N His-Leu-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O TWROVBNEHJSXDG-IHRRRGAJSA-N 0.000 description 1
- CKRJBQJIGOEKMC-SRVKXCTJSA-N His-Lys-Ser Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O CKRJBQJIGOEKMC-SRVKXCTJSA-N 0.000 description 1
- FHKZHRMERJUXRJ-DCAQKATOSA-N His-Ser-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CN=CN1 FHKZHRMERJUXRJ-DCAQKATOSA-N 0.000 description 1
- ZHHLTWUOWXHVQJ-YUMQZZPRSA-N His-Ser-Gly Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)NCC(=O)O)N ZHHLTWUOWXHVQJ-YUMQZZPRSA-N 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 101000617738 Homo sapiens Survival motor neuron protein Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- WUEIUSDAECDLQO-NAKRPEOUSA-N Ile-Ala-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)O)N WUEIUSDAECDLQO-NAKRPEOUSA-N 0.000 description 1
- YOTNPRLPIPHQSB-XUXIUFHCSA-N Ile-Arg-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N YOTNPRLPIPHQSB-XUXIUFHCSA-N 0.000 description 1
- HDODQNPMSHDXJT-GHCJXIJMSA-N Ile-Asn-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O HDODQNPMSHDXJT-GHCJXIJMSA-N 0.000 description 1
- DMZOUKXXHJQPTL-GRLWGSQLSA-N Ile-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N DMZOUKXXHJQPTL-GRLWGSQLSA-N 0.000 description 1
- YBJWJQQBWRARLT-KBIXCLLPSA-N Ile-Gln-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O YBJWJQQBWRARLT-KBIXCLLPSA-N 0.000 description 1
- PHIXPNQDGGILMP-YVNDNENWSA-N Ile-Glu-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N PHIXPNQDGGILMP-YVNDNENWSA-N 0.000 description 1
- IGJWJGIHUFQANP-LAEOZQHASA-N Ile-Gly-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)O)N IGJWJGIHUFQANP-LAEOZQHASA-N 0.000 description 1
- NYEYYMLUABXDMC-NHCYSSNCSA-N Ile-Gly-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)O)N NYEYYMLUABXDMC-NHCYSSNCSA-N 0.000 description 1
- LWWILHPVAKKLQS-QXEWZRGKSA-N Ile-Gly-Met Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CCSC)C(=O)O)N LWWILHPVAKKLQS-QXEWZRGKSA-N 0.000 description 1
- SJLVSMMIFYTSGY-GRLWGSQLSA-N Ile-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N SJLVSMMIFYTSGY-GRLWGSQLSA-N 0.000 description 1
- DMSVBUWGDLYNLC-IAVJCBSLSA-N Ile-Ile-Phe Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 DMSVBUWGDLYNLC-IAVJCBSLSA-N 0.000 description 1
- TWYOYAKMLHWMOJ-ZPFDUUQYSA-N Ile-Leu-Asn Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O TWYOYAKMLHWMOJ-ZPFDUUQYSA-N 0.000 description 1
- DBXXASNNDTXOLU-MXAVVETBSA-N Ile-Leu-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N DBXXASNNDTXOLU-MXAVVETBSA-N 0.000 description 1
- HPCFRQWLTRDGHT-AJNGGQMLSA-N Ile-Leu-Leu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O HPCFRQWLTRDGHT-AJNGGQMLSA-N 0.000 description 1
- PHRWFSFCNJPWRO-PPCPHDFISA-N Ile-Leu-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N PHRWFSFCNJPWRO-PPCPHDFISA-N 0.000 description 1
- FJWALBCCVIHZBS-QXEWZRGKSA-N Ile-Met-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)O)N FJWALBCCVIHZBS-QXEWZRGKSA-N 0.000 description 1
- FTUZWJVSNZMLPI-RVMXOQNASA-N Ile-Met-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N1CCC[C@@H]1C(=O)O)N FTUZWJVSNZMLPI-RVMXOQNASA-N 0.000 description 1
- NPAYJTAXWXJKLO-NAKRPEOUSA-N Ile-Met-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)O)N NPAYJTAXWXJKLO-NAKRPEOUSA-N 0.000 description 1
- SAVXZJYTTQQQDD-QEWYBTABSA-N Ile-Phe-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N SAVXZJYTTQQQDD-QEWYBTABSA-N 0.000 description 1
- XLXPYSDGMXTTNQ-UHFFFAOYSA-N Ile-Phe-Leu Natural products CCC(C)C(N)C(=O)NC(C(=O)NC(CC(C)C)C(O)=O)CC1=CC=CC=C1 XLXPYSDGMXTTNQ-UHFFFAOYSA-N 0.000 description 1
- VEPIBPGLTLPBDW-URLPEUOOSA-N Ile-Phe-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N VEPIBPGLTLPBDW-URLPEUOOSA-N 0.000 description 1
- ZNOBVZFCHNHKHA-KBIXCLLPSA-N Ile-Ser-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ZNOBVZFCHNHKHA-KBIXCLLPSA-N 0.000 description 1
- AGGIYSLVUKVOPT-HTFCKZLJSA-N Ile-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N AGGIYSLVUKVOPT-HTFCKZLJSA-N 0.000 description 1
- ZDNNDIJTUHQCAM-MXAVVETBSA-N Ile-Ser-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N ZDNNDIJTUHQCAM-MXAVVETBSA-N 0.000 description 1
- WCNWGAUZWWSYDG-SVSWQMSJSA-N Ile-Thr-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)O)N WCNWGAUZWWSYDG-SVSWQMSJSA-N 0.000 description 1
- QHUREMVLLMNUAX-OSUNSFLBSA-N Ile-Thr-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)O)N QHUREMVLLMNUAX-OSUNSFLBSA-N 0.000 description 1
- VBGCPJBKUXRYDA-DSYPUSFNSA-N Ile-Trp-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCCCN)C(=O)O)N VBGCPJBKUXRYDA-DSYPUSFNSA-N 0.000 description 1
- ZGKVPOSSTGHJAF-HJPIBITLSA-N Ile-Tyr-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CO)C(=O)O)N ZGKVPOSSTGHJAF-HJPIBITLSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 108010065920 Insulin Lispro Proteins 0.000 description 1
- 108010066420 Iron-Regulatory Proteins Proteins 0.000 description 1
- 102000018434 Iron-Regulatory Proteins Human genes 0.000 description 1
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 1
- HGCNKOLVKRAVHD-UHFFFAOYSA-N L-Met-L-Phe Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 HGCNKOLVKRAVHD-UHFFFAOYSA-N 0.000 description 1
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 1
- SITWEMZOJNKJCH-UHFFFAOYSA-N L-alanine-L-arginine Natural products CC(N)C(=O)NC(C(O)=O)CCCNC(N)=N SITWEMZOJNKJCH-UHFFFAOYSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- RCFDOSNHHZGBOY-UHFFFAOYSA-N L-isoleucyl-L-alanine Natural products CCC(C)C(N)C(=O)NC(C)C(O)=O RCFDOSNHHZGBOY-UHFFFAOYSA-N 0.000 description 1
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 1
- KFKWRHQBZQICHA-STQMWFEESA-N L-leucyl-L-phenylalanine Natural products CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KFKWRHQBZQICHA-STQMWFEESA-N 0.000 description 1
- TYYLDKGBCJGJGW-UHFFFAOYSA-N L-tryptophan-L-tyrosine Natural products C=1NC2=CC=CC=C2C=1CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 TYYLDKGBCJGJGW-UHFFFAOYSA-N 0.000 description 1
- UBORTCNDUKBEOP-UHFFFAOYSA-N L-xanthosine Natural products OC1C(O)C(CO)OC1N1C(NC(=O)NC2=O)=C2N=C1 UBORTCNDUKBEOP-UHFFFAOYSA-N 0.000 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 1
- 241000270322 Lepidosauria Species 0.000 description 1
- BQSLGJHIAGOZCD-CIUDSAMLSA-N Leu-Ala-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O BQSLGJHIAGOZCD-CIUDSAMLSA-N 0.000 description 1
- YOZCKMXHBYKOMQ-IHRRRGAJSA-N Leu-Arg-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N YOZCKMXHBYKOMQ-IHRRRGAJSA-N 0.000 description 1
- GPXFZVUVPCFTMG-AVGNSLFASA-N Leu-Arg-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(C)C GPXFZVUVPCFTMG-AVGNSLFASA-N 0.000 description 1
- UCOCBWDBHCUPQP-DCAQKATOSA-N Leu-Arg-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O UCOCBWDBHCUPQP-DCAQKATOSA-N 0.000 description 1
- XYUBOFCTGPZFSA-WDSOQIARSA-N Leu-Arg-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 XYUBOFCTGPZFSA-WDSOQIARSA-N 0.000 description 1
- CUXRXAIAVYLVFD-ULQDDVLXSA-N Leu-Arg-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 CUXRXAIAVYLVFD-ULQDDVLXSA-N 0.000 description 1
- IGUOAYLTQJLPPD-DCAQKATOSA-N Leu-Asn-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N IGUOAYLTQJLPPD-DCAQKATOSA-N 0.000 description 1
- WGNOPSQMIQERPK-UHFFFAOYSA-N Leu-Asn-Pro Natural products CC(C)CC(N)C(=O)NC(CC(=O)N)C(=O)N1CCCC1C(=O)O WGNOPSQMIQERPK-UHFFFAOYSA-N 0.000 description 1
- FIJMQLGQLBLBOL-HJGDQZAQSA-N Leu-Asn-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FIJMQLGQLBLBOL-HJGDQZAQSA-N 0.000 description 1
- DLFAACQHIRSQGG-CIUDSAMLSA-N Leu-Asp-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O DLFAACQHIRSQGG-CIUDSAMLSA-N 0.000 description 1
- ILJREDZFPHTUIE-GUBZILKMSA-N Leu-Asp-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ILJREDZFPHTUIE-GUBZILKMSA-N 0.000 description 1
- MYGQXVYRZMKRDB-SRVKXCTJSA-N Leu-Asp-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN MYGQXVYRZMKRDB-SRVKXCTJSA-N 0.000 description 1
- KWURTLAFFDOTEQ-GUBZILKMSA-N Leu-Cys-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N KWURTLAFFDOTEQ-GUBZILKMSA-N 0.000 description 1
- VQPPIMUZCZCOIL-GUBZILKMSA-N Leu-Gln-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O VQPPIMUZCZCOIL-GUBZILKMSA-N 0.000 description 1
- KAFOIVJDVSZUMD-DCAQKATOSA-N Leu-Gln-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O KAFOIVJDVSZUMD-DCAQKATOSA-N 0.000 description 1
- KAFOIVJDVSZUMD-UHFFFAOYSA-N Leu-Gln-Gln Natural products CC(C)CC(N)C(=O)NC(CCC(N)=O)C(=O)NC(CCC(N)=O)C(O)=O KAFOIVJDVSZUMD-UHFFFAOYSA-N 0.000 description 1
- FQZPTCNSNPWHLJ-AVGNSLFASA-N Leu-Gln-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(O)=O FQZPTCNSNPWHLJ-AVGNSLFASA-N 0.000 description 1
- CQGSYZCULZMEDE-UHFFFAOYSA-N Leu-Gln-Pro Natural products CC(C)CC(N)C(=O)NC(CCC(N)=O)C(=O)N1CCCC1C(O)=O CQGSYZCULZMEDE-UHFFFAOYSA-N 0.000 description 1
- WMTOVWLLDGQGCV-GUBZILKMSA-N Leu-Glu-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CS)C(=O)O)N WMTOVWLLDGQGCV-GUBZILKMSA-N 0.000 description 1
- OXRLYTYUXAQTHP-YUMQZZPRSA-N Leu-Gly-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(O)=O OXRLYTYUXAQTHP-YUMQZZPRSA-N 0.000 description 1
- BABSVXFGKFLIGW-UWVGGRQHSA-N Leu-Gly-Arg Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCNC(N)=N BABSVXFGKFLIGW-UWVGGRQHSA-N 0.000 description 1
- FMEICTQWUKNAGC-YUMQZZPRSA-N Leu-Gly-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O FMEICTQWUKNAGC-YUMQZZPRSA-N 0.000 description 1
- HYMLKESRWLZDBR-WEDXCCLWSA-N Leu-Gly-Thr Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HYMLKESRWLZDBR-WEDXCCLWSA-N 0.000 description 1
- VZBIUJURDLFFOE-IHRRRGAJSA-N Leu-His-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VZBIUJURDLFFOE-IHRRRGAJSA-N 0.000 description 1
- OHZIZVWQXJPBJS-IXOXFDKPSA-N Leu-His-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OHZIZVWQXJPBJS-IXOXFDKPSA-N 0.000 description 1
- QLDHBYRUNQZIJQ-DKIMLUQUSA-N Leu-Ile-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QLDHBYRUNQZIJQ-DKIMLUQUSA-N 0.000 description 1
- LIINDKYIGYTDLG-PPCPHDFISA-N Leu-Ile-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LIINDKYIGYTDLG-PPCPHDFISA-N 0.000 description 1
- IAJFFZORSWOZPQ-SRVKXCTJSA-N Leu-Leu-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IAJFFZORSWOZPQ-SRVKXCTJSA-N 0.000 description 1
- YOKVEHGYYQEQOP-QWRGUYRKSA-N Leu-Leu-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YOKVEHGYYQEQOP-QWRGUYRKSA-N 0.000 description 1
- RXGLHDWAZQECBI-SRVKXCTJSA-N Leu-Leu-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O RXGLHDWAZQECBI-SRVKXCTJSA-N 0.000 description 1
- ZRHDPZAAWLXXIR-SRVKXCTJSA-N Leu-Lys-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O ZRHDPZAAWLXXIR-SRVKXCTJSA-N 0.000 description 1
- HVHRPWQEQHIQJF-AVGNSLFASA-N Leu-Lys-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O HVHRPWQEQHIQJF-AVGNSLFASA-N 0.000 description 1
- PKKMDPNFGULLNQ-AVGNSLFASA-N Leu-Met-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O PKKMDPNFGULLNQ-AVGNSLFASA-N 0.000 description 1
- FLNPJLDPGMLWAU-UWVGGRQHSA-N Leu-Met-Gly Chemical compound OC(=O)CNC(=O)[C@H](CCSC)NC(=O)[C@@H](N)CC(C)C FLNPJLDPGMLWAU-UWVGGRQHSA-N 0.000 description 1
- JVTYXRRFZCEPPK-RHYQMDGZSA-N Leu-Met-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)N)O JVTYXRRFZCEPPK-RHYQMDGZSA-N 0.000 description 1
- NJMXCOOEFLMZSR-AVGNSLFASA-N Leu-Met-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C(C)C)C(O)=O NJMXCOOEFLMZSR-AVGNSLFASA-N 0.000 description 1
- QMKFDEUJGYNFMC-AVGNSLFASA-N Leu-Pro-Arg Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O QMKFDEUJGYNFMC-AVGNSLFASA-N 0.000 description 1
- KWLWZYMNUZJKMZ-IHRRRGAJSA-N Leu-Pro-Leu Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O KWLWZYMNUZJKMZ-IHRRRGAJSA-N 0.000 description 1
- JIHDFWWRYHSAQB-GUBZILKMSA-N Leu-Ser-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O JIHDFWWRYHSAQB-GUBZILKMSA-N 0.000 description 1
- RGUXWMDNCPMQFB-YUMQZZPRSA-N Leu-Ser-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RGUXWMDNCPMQFB-YUMQZZPRSA-N 0.000 description 1
- ADJWHHZETYAAAX-SRVKXCTJSA-N Leu-Ser-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N ADJWHHZETYAAAX-SRVKXCTJSA-N 0.000 description 1
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 1
- PPGBXYKMUMHFBF-KATARQTJSA-N Leu-Ser-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PPGBXYKMUMHFBF-KATARQTJSA-N 0.000 description 1
- ZDJQVSIPFLMNOX-RHYQMDGZSA-N Leu-Thr-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N ZDJQVSIPFLMNOX-RHYQMDGZSA-N 0.000 description 1
- GZRABTMNWJXFMH-UVOCVTCTSA-N Leu-Thr-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZRABTMNWJXFMH-UVOCVTCTSA-N 0.000 description 1
- FPFOYSCDUWTZBF-IHPCNDPISA-N Leu-Trp-Leu Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H]([NH3+])CC(C)C)C(=O)N[C@@H](CC(C)C)C([O-])=O)=CNC2=C1 FPFOYSCDUWTZBF-IHPCNDPISA-N 0.000 description 1
- VKVDRTGWLVZJOM-DCAQKATOSA-N Leu-Val-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O VKVDRTGWLVZJOM-DCAQKATOSA-N 0.000 description 1
- QESXLSQLQHHTIX-RHYQMDGZSA-N Leu-Val-Thr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QESXLSQLQHHTIX-RHYQMDGZSA-N 0.000 description 1
- 240000006240 Linum usitatissimum Species 0.000 description 1
- 235000004431 Linum usitatissimum Nutrition 0.000 description 1
- 208000000501 Lipidoses Diseases 0.000 description 1
- 206010024585 Lipidosis Diseases 0.000 description 1
- 239000012097 Lipofectamine 2000 Substances 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- FZIJIFCXUCZHOL-CIUDSAMLSA-N Lys-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN FZIJIFCXUCZHOL-CIUDSAMLSA-N 0.000 description 1
- MPGHETGWWWUHPY-CIUDSAMLSA-N Lys-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN MPGHETGWWWUHPY-CIUDSAMLSA-N 0.000 description 1
- VHFFQUSNFFIZBT-CIUDSAMLSA-N Lys-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCCCN)N VHFFQUSNFFIZBT-CIUDSAMLSA-N 0.000 description 1
- WSXTWLJHTLRFLW-SRVKXCTJSA-N Lys-Ala-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O WSXTWLJHTLRFLW-SRVKXCTJSA-N 0.000 description 1
- VHXMZJGOKIMETG-CQDKDKBSSA-N Lys-Ala-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CCCCN)N VHXMZJGOKIMETG-CQDKDKBSSA-N 0.000 description 1
- HQVDJTYKCMIWJP-YUMQZZPRSA-N Lys-Asn-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O HQVDJTYKCMIWJP-YUMQZZPRSA-N 0.000 description 1
- NCTDKZKNBDZDOL-GARJFASQSA-N Lys-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCCN)N)C(=O)O NCTDKZKNBDZDOL-GARJFASQSA-N 0.000 description 1
- DGWXCIORNLWGGG-CIUDSAMLSA-N Lys-Asn-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O DGWXCIORNLWGGG-CIUDSAMLSA-N 0.000 description 1
- WGCKDDHUFPQSMZ-ZPFDUUQYSA-N Lys-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCCCN WGCKDDHUFPQSMZ-ZPFDUUQYSA-N 0.000 description 1
- IWWMPCPLFXFBAF-SRVKXCTJSA-N Lys-Asp-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O IWWMPCPLFXFBAF-SRVKXCTJSA-N 0.000 description 1
- YFGWNAROEYWGNL-GUBZILKMSA-N Lys-Gln-Asn Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O YFGWNAROEYWGNL-GUBZILKMSA-N 0.000 description 1
- ZXEUFAVXODIPHC-GUBZILKMSA-N Lys-Glu-Asn Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O ZXEUFAVXODIPHC-GUBZILKMSA-N 0.000 description 1
- GQFDWEDHOQRNLC-QWRGUYRKSA-N Lys-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN GQFDWEDHOQRNLC-QWRGUYRKSA-N 0.000 description 1
- ZXFRGTAIIZHNHG-AJNGGQMLSA-N Lys-Ile-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)O)NC(=O)[C@H](CCCCN)N ZXFRGTAIIZHNHG-AJNGGQMLSA-N 0.000 description 1
- QOJDBRUCOXQSSK-AJNGGQMLSA-N Lys-Ile-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(O)=O QOJDBRUCOXQSSK-AJNGGQMLSA-N 0.000 description 1
- SKRGVGLIRUGANF-AVGNSLFASA-N Lys-Leu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SKRGVGLIRUGANF-AVGNSLFASA-N 0.000 description 1
- WVJNGSFKBKOKRV-AJNGGQMLSA-N Lys-Leu-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WVJNGSFKBKOKRV-AJNGGQMLSA-N 0.000 description 1
- AIRZWUMAHCDDHR-KKUMJFAQSA-N Lys-Leu-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O AIRZWUMAHCDDHR-KKUMJFAQSA-N 0.000 description 1
- ORVFEGYUJITPGI-IHRRRGAJSA-N Lys-Leu-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCCCN ORVFEGYUJITPGI-IHRRRGAJSA-N 0.000 description 1
- VSTNAUBHKQPVJX-IHRRRGAJSA-N Lys-Met-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O VSTNAUBHKQPVJX-IHRRRGAJSA-N 0.000 description 1
- ALEVUGKHINJNIF-QEJZJMRPSA-N Lys-Phe-Ala Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=CC=C1 ALEVUGKHINJNIF-QEJZJMRPSA-N 0.000 description 1
- LOGFVTREOLYCPF-RHYQMDGZSA-N Lys-Pro-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-RHYQMDGZSA-N 0.000 description 1
- MIROMRNASYKZNL-ULQDDVLXSA-N Lys-Pro-Tyr Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 MIROMRNASYKZNL-ULQDDVLXSA-N 0.000 description 1
- ZUGVARDEGWMMLK-SRVKXCTJSA-N Lys-Ser-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN ZUGVARDEGWMMLK-SRVKXCTJSA-N 0.000 description 1
- MIFFFXHMAHFACR-KATARQTJSA-N Lys-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CCCCN MIFFFXHMAHFACR-KATARQTJSA-N 0.000 description 1
- RPWTZTBIFGENIA-VOAKCMCISA-N Lys-Thr-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O RPWTZTBIFGENIA-VOAKCMCISA-N 0.000 description 1
- RMOKGALPSPOYKE-KATARQTJSA-N Lys-Thr-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O RMOKGALPSPOYKE-KATARQTJSA-N 0.000 description 1
- VWJFOUBDZIUXGA-AVGNSLFASA-N Lys-Val-Met Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CCCCN)N VWJFOUBDZIUXGA-AVGNSLFASA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-L Malonate Chemical compound [O-]C(=O)CC([O-])=O OFOBLEOULBTSOW-UHFFFAOYSA-L 0.000 description 1
- ACYHZNZHIZWLQF-BQBZGAKWSA-N Met-Asn-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O ACYHZNZHIZWLQF-BQBZGAKWSA-N 0.000 description 1
- PTYVBBNIAQWUFV-DCAQKATOSA-N Met-Cys-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CCSC)N PTYVBBNIAQWUFV-DCAQKATOSA-N 0.000 description 1
- OXHSZBRPUGNMKW-DCAQKATOSA-N Met-Gln-Arg Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O OXHSZBRPUGNMKW-DCAQKATOSA-N 0.000 description 1
- AETNZPKUUYYYEK-CIUDSAMLSA-N Met-Glu-Asn Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O AETNZPKUUYYYEK-CIUDSAMLSA-N 0.000 description 1
- LQMHZERGCQJKAH-STQMWFEESA-N Met-Gly-Phe Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 LQMHZERGCQJKAH-STQMWFEESA-N 0.000 description 1
- ORRNBLTZBBESPN-HJWJTTGWSA-N Met-Ile-Phe Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 ORRNBLTZBBESPN-HJWJTTGWSA-N 0.000 description 1
- CRVSHEPROQHVQT-AVGNSLFASA-N Met-Met-Lys Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)O)N CRVSHEPROQHVQT-AVGNSLFASA-N 0.000 description 1
- LUYURUYVNYGKGM-RCWTZXSCSA-N Met-Pro-Thr Chemical compound CSCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O LUYURUYVNYGKGM-RCWTZXSCSA-N 0.000 description 1
- DBMLDOWSVHMQQN-XGEHTFHBSA-N Met-Ser-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O DBMLDOWSVHMQQN-XGEHTFHBSA-N 0.000 description 1
- ZBLSZPYQQRIHQU-RCWTZXSCSA-N Met-Thr-Val Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O ZBLSZPYQQRIHQU-RCWTZXSCSA-N 0.000 description 1
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 1
- 241000721550 Micromys minutus Species 0.000 description 1
- 241001508687 Mustela erminea Species 0.000 description 1
- 241000282341 Mustela putorius furo Species 0.000 description 1
- SGSSKEDGVONRGC-UHFFFAOYSA-N N(2)-methylguanine Chemical compound O=C1NC(NC)=NC2=C1N=CN2 SGSSKEDGVONRGC-UHFFFAOYSA-N 0.000 description 1
- VQAYFKKCNSOZKM-IOSLPCCCSA-N N(6)-methyladenosine Chemical compound C1=NC=2C(NC)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O VQAYFKKCNSOZKM-IOSLPCCCSA-N 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- XZFYRXDAULDNFX-UHFFFAOYSA-N N-L-cysteinyl-L-phenylalanine Natural products SCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XZFYRXDAULDNFX-UHFFFAOYSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- VQAYFKKCNSOZKM-UHFFFAOYSA-N NSC 29409 Natural products C1=NC=2C(NC)=NC=NC=2N1C1OC(CO)C(O)C1O VQAYFKKCNSOZKM-UHFFFAOYSA-N 0.000 description 1
- BQVUABVGYYSDCJ-UHFFFAOYSA-N Nalpha-L-Leucyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)CC(C)C)C(O)=O)=CNC2=C1 BQVUABVGYYSDCJ-UHFFFAOYSA-N 0.000 description 1
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 1
- 108010089610 Nuclear Proteins Proteins 0.000 description 1
- 102000007999 Nuclear Proteins Human genes 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- REYJJPSVUYRZGE-UHFFFAOYSA-N Octadecylamine Chemical class CCCCCCCCCCCCCCCCCCN REYJJPSVUYRZGE-UHFFFAOYSA-N 0.000 description 1
- 241000276701 Oreochromis mossambicus Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- TZRXHJWUDPFEEY-UHFFFAOYSA-N Pentaerythritol Tetranitrate Chemical compound [O-][N+](=O)OCC(CO[N+]([O-])=O)(CO[N+]([O-])=O)CO[N+]([O-])=O TZRXHJWUDPFEEY-UHFFFAOYSA-N 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108091093037 Peptide nucleic acid Proteins 0.000 description 1
- CGOMLCQJEMWMCE-STQMWFEESA-N Phe-Arg-Gly Chemical compound NC(N)=NCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 CGOMLCQJEMWMCE-STQMWFEESA-N 0.000 description 1
- YYRCPTVAPLQRNC-ULQDDVLXSA-N Phe-Arg-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CC1=CC=CC=C1 YYRCPTVAPLQRNC-ULQDDVLXSA-N 0.000 description 1
- CDNPIRSCAFMMBE-SRVKXCTJSA-N Phe-Asn-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O CDNPIRSCAFMMBE-SRVKXCTJSA-N 0.000 description 1
- UEEVBGHEGJMDDV-AVGNSLFASA-N Phe-Asp-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 UEEVBGHEGJMDDV-AVGNSLFASA-N 0.000 description 1
- WFDAEEUZPZSMOG-SRVKXCTJSA-N Phe-Cys-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(O)=O WFDAEEUZPZSMOG-SRVKXCTJSA-N 0.000 description 1
- YEEFZOKPYOUXMX-KKUMJFAQSA-N Phe-Gln-Met Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O YEEFZOKPYOUXMX-KKUMJFAQSA-N 0.000 description 1
- YZJKNDCEPDDIDA-BZSNNMDCSA-N Phe-His-Lys Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC=CC=1)C1=CN=CN1 YZJKNDCEPDDIDA-BZSNNMDCSA-N 0.000 description 1
- VZFPYFRVHMSSNA-JURCDPSOSA-N Phe-Ile-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=CC=C1 VZFPYFRVHMSSNA-JURCDPSOSA-N 0.000 description 1
- GYEPCBNTTRORKW-PCBIJLKTSA-N Phe-Ile-Asp Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O GYEPCBNTTRORKW-PCBIJLKTSA-N 0.000 description 1
- BWTKUQPNOMMKMA-FIRPJDEBSA-N Phe-Ile-Phe Chemical compound C([C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 BWTKUQPNOMMKMA-FIRPJDEBSA-N 0.000 description 1
- OHIYMVFLQXTZAW-UFYCRDLUSA-N Phe-Met-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O OHIYMVFLQXTZAW-UFYCRDLUSA-N 0.000 description 1
- KAJLHCWRWDSROH-BZSNNMDCSA-N Phe-Phe-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=CC=C1 KAJLHCWRWDSROH-BZSNNMDCSA-N 0.000 description 1
- YMTMNYNEZDAGMW-RNXOBYDBSA-N Phe-Phe-Trp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N[C@@H](CC3=CNC4=CC=CC=C43)C(=O)O)N YMTMNYNEZDAGMW-RNXOBYDBSA-N 0.000 description 1
- GZGPMBKUJDRICD-ULQDDVLXSA-N Phe-Pro-His Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)N)C(=O)N[C@@H](CC3=CN=CN3)C(=O)O GZGPMBKUJDRICD-ULQDDVLXSA-N 0.000 description 1
- AFNJAQVMTIQTCB-DLOVCJGASA-N Phe-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=CC=C1 AFNJAQVMTIQTCB-DLOVCJGASA-N 0.000 description 1
- JXQVYPWVGUOIDV-MXAVVETBSA-N Phe-Ser-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JXQVYPWVGUOIDV-MXAVVETBSA-N 0.000 description 1
- MRWOVVNKSXXLRP-IHPCNDPISA-N Phe-Ser-Trp Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O MRWOVVNKSXXLRP-IHPCNDPISA-N 0.000 description 1
- YFXXRYFWJFQAFW-JHYOHUSXSA-N Phe-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N)O YFXXRYFWJFQAFW-JHYOHUSXSA-N 0.000 description 1
- PCNDJXKNXGMECE-UHFFFAOYSA-N Phenazine Chemical group C1=CC=CC2=NC3=CC=CC=C3N=C21 PCNDJXKNXGMECE-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- ZCXQTRXYZOSGJR-FXQIFTODSA-N Pro-Asp-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O ZCXQTRXYZOSGJR-FXQIFTODSA-N 0.000 description 1
- WGAQWMRJUFQXMF-ZPFDUUQYSA-N Pro-Gln-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WGAQWMRJUFQXMF-ZPFDUUQYSA-N 0.000 description 1
- POQFNPILEQEODH-FXQIFTODSA-N Pro-Ser-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O POQFNPILEQEODH-FXQIFTODSA-N 0.000 description 1
- FNGOXVQBBCMFKV-CIUDSAMLSA-N Pro-Ser-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O FNGOXVQBBCMFKV-CIUDSAMLSA-N 0.000 description 1
- FDMCIBSQRKFSTJ-RHYQMDGZSA-N Pro-Thr-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O FDMCIBSQRKFSTJ-RHYQMDGZSA-N 0.000 description 1
- FZXSYIPVAFVYBH-KKUMJFAQSA-N Pro-Tyr-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O FZXSYIPVAFVYBH-KKUMJFAQSA-N 0.000 description 1
- DYJTXTCEXMCPBF-UFYCRDLUSA-N Pro-Tyr-Phe Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CC3=CC=CC=C3)C(=O)O DYJTXTCEXMCPBF-UFYCRDLUSA-N 0.000 description 1
- KHRLUIPIMIQFGT-AVGNSLFASA-N Pro-Val-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O KHRLUIPIMIQFGT-AVGNSLFASA-N 0.000 description 1
- FIODMZKLZFLYQP-GUBZILKMSA-N Pro-Val-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O FIODMZKLZFLYQP-GUBZILKMSA-N 0.000 description 1
- YDTUEBLEAVANFH-RCWTZXSCSA-N Pro-Val-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 YDTUEBLEAVANFH-RCWTZXSCSA-N 0.000 description 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 102000009609 Pyrophosphatases Human genes 0.000 description 1
- 108010009413 Pyrophosphatases Proteins 0.000 description 1
- 108010065868 RNA polymerase SP6 Proteins 0.000 description 1
- 108091028664 Ribonucleotide Proteins 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 241000220287 Sedum rubrotinctum Species 0.000 description 1
- LVVBAKCGXXUHFO-ZLUOBGJFSA-N Ser-Ala-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(O)=O LVVBAKCGXXUHFO-ZLUOBGJFSA-N 0.000 description 1
- CNIIKZQXBBQHCX-FXQIFTODSA-N Ser-Asp-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O CNIIKZQXBBQHCX-FXQIFTODSA-N 0.000 description 1
- SWIQQMYVHIXPEK-FXQIFTODSA-N Ser-Cys-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O SWIQQMYVHIXPEK-FXQIFTODSA-N 0.000 description 1
- YRBGKVIWMNEVCZ-WDSKDSINSA-N Ser-Glu-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O YRBGKVIWMNEVCZ-WDSKDSINSA-N 0.000 description 1
- LALNXSXEYFUUDD-GUBZILKMSA-N Ser-Glu-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LALNXSXEYFUUDD-GUBZILKMSA-N 0.000 description 1
- GRSLLFZTTLBOQX-CIUDSAMLSA-N Ser-Glu-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CO)N GRSLLFZTTLBOQX-CIUDSAMLSA-N 0.000 description 1
- SFTZWNJFZYOLBD-ZDLURKLDSA-N Ser-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO SFTZWNJFZYOLBD-ZDLURKLDSA-N 0.000 description 1
- BKZYBLLIBOBOOW-GHCJXIJMSA-N Ser-Ile-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O BKZYBLLIBOBOOW-GHCJXIJMSA-N 0.000 description 1
- RIAKPZVSNBBNRE-BJDJZHNGSA-N Ser-Ile-Leu Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O RIAKPZVSNBBNRE-BJDJZHNGSA-N 0.000 description 1
- MOINZPRHJGTCHZ-MMWGEVLESA-N Ser-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N MOINZPRHJGTCHZ-MMWGEVLESA-N 0.000 description 1
- ZOPISOXXPQNOCO-SVSWQMSJSA-N Ser-Ile-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CO)N ZOPISOXXPQNOCO-SVSWQMSJSA-N 0.000 description 1
- FUMGHWDRRFCKEP-CIUDSAMLSA-N Ser-Leu-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O FUMGHWDRRFCKEP-CIUDSAMLSA-N 0.000 description 1
- VZQRNAYURWAEFE-KKUMJFAQSA-N Ser-Leu-Phe Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 VZQRNAYURWAEFE-KKUMJFAQSA-N 0.000 description 1
- IXZHZUGGKLRHJD-DCAQKATOSA-N Ser-Leu-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O IXZHZUGGKLRHJD-DCAQKATOSA-N 0.000 description 1
- PPNPDKGQRFSCAC-CIUDSAMLSA-N Ser-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPNPDKGQRFSCAC-CIUDSAMLSA-N 0.000 description 1
- JLPMFVAIQHCBDC-CIUDSAMLSA-N Ser-Lys-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CO)N JLPMFVAIQHCBDC-CIUDSAMLSA-N 0.000 description 1
- LRWBCWGEUCKDTN-BJDJZHNGSA-N Ser-Lys-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LRWBCWGEUCKDTN-BJDJZHNGSA-N 0.000 description 1
- XUDRHBPSPAPDJP-SRVKXCTJSA-N Ser-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CO XUDRHBPSPAPDJP-SRVKXCTJSA-N 0.000 description 1
- OCWWJBZQXGYQCA-DCAQKATOSA-N Ser-Lys-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(O)=O OCWWJBZQXGYQCA-DCAQKATOSA-N 0.000 description 1
- MQUZANJDFOQOBX-SRVKXCTJSA-N Ser-Phe-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O MQUZANJDFOQOBX-SRVKXCTJSA-N 0.000 description 1
- FKYWFUYPVKLJLP-DCAQKATOSA-N Ser-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO FKYWFUYPVKLJLP-DCAQKATOSA-N 0.000 description 1
- OZPDGESCTGGNAD-CIUDSAMLSA-N Ser-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CO OZPDGESCTGGNAD-CIUDSAMLSA-N 0.000 description 1
- UQGAAZXSCGWMFU-UBHSHLNASA-N Ser-Trp-Asp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CO)N UQGAAZXSCGWMFU-UBHSHLNASA-N 0.000 description 1
- UKKROEYWYIHWBD-ZKWXMUAHSA-N Ser-Val-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O UKKROEYWYIHWBD-ZKWXMUAHSA-N 0.000 description 1
- JGUWRQWULDWNCM-FXQIFTODSA-N Ser-Val-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O JGUWRQWULDWNCM-FXQIFTODSA-N 0.000 description 1
- HSWXBJCBYSWBPT-GUBZILKMSA-N Ser-Val-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)C(O)=O HSWXBJCBYSWBPT-GUBZILKMSA-N 0.000 description 1
- 229910004298 SiO 2 Inorganic materials 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 102100021947 Survival motor neuron protein Human genes 0.000 description 1
- 101710137500 T7 RNA polymerase Proteins 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- ZUXQFMVPAYGPFJ-JXUBOQSCSA-N Thr-Ala-Lys Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN ZUXQFMVPAYGPFJ-JXUBOQSCSA-N 0.000 description 1
- DWYAUVCQDTZIJI-VZFHVOOUSA-N Thr-Ala-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DWYAUVCQDTZIJI-VZFHVOOUSA-N 0.000 description 1
- MQBTXMPQNCGSSZ-OSUNSFLBSA-N Thr-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@@H](C)O)CCCN=C(N)N MQBTXMPQNCGSSZ-OSUNSFLBSA-N 0.000 description 1
- NAXBBCLCEOTAIG-RHYQMDGZSA-N Thr-Arg-Lys Chemical compound NC(N)=NCCC[C@H](NC(=O)[C@@H](N)[C@H](O)C)C(=O)N[C@@H](CCCCN)C(O)=O NAXBBCLCEOTAIG-RHYQMDGZSA-N 0.000 description 1
- GZYNMZQXFRWDFH-YTWAJWBKSA-N Thr-Arg-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@@H]1C(=O)O)N)O GZYNMZQXFRWDFH-YTWAJWBKSA-N 0.000 description 1
- XOTBWOCSLMBGMF-SUSMZKCASA-N Thr-Glu-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XOTBWOCSLMBGMF-SUSMZKCASA-N 0.000 description 1
- DJDSEDOKJTZBAR-ZDLURKLDSA-N Thr-Gly-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O DJDSEDOKJTZBAR-ZDLURKLDSA-N 0.000 description 1
- UYTYTDMCDBPDSC-URLPEUOOSA-N Thr-Ile-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N UYTYTDMCDBPDSC-URLPEUOOSA-N 0.000 description 1
- MEJHFIOYJHTWMK-VOAKCMCISA-N Thr-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)[C@@H](C)O MEJHFIOYJHTWMK-VOAKCMCISA-N 0.000 description 1
- MECLEFZMPPOEAC-VOAKCMCISA-N Thr-Leu-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MECLEFZMPPOEAC-VOAKCMCISA-N 0.000 description 1
- MGJLBZFUXUGMML-VOAKCMCISA-N Thr-Lys-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MGJLBZFUXUGMML-VOAKCMCISA-N 0.000 description 1
- WPSKTVVMQCXPRO-BWBBJGPYSA-N Thr-Ser-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O WPSKTVVMQCXPRO-BWBBJGPYSA-N 0.000 description 1
- COYHRQWNJDJCNA-NUJDXYNKSA-N Thr-Thr-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O COYHRQWNJDJCNA-NUJDXYNKSA-N 0.000 description 1
- BJJRNAVDQGREGC-HOUAVDHOSA-N Thr-Trp-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O BJJRNAVDQGREGC-HOUAVDHOSA-N 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 108090000992 Transferases Proteins 0.000 description 1
- 102000004357 Transferases Human genes 0.000 description 1
- BIJDDZBDSJLWJY-PJODQICGSA-N Trp-Ala-Val Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O BIJDDZBDSJLWJY-PJODQICGSA-N 0.000 description 1
- MVHHTXAUJCIOMZ-WDSOQIARSA-N Trp-Arg-Lys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N MVHHTXAUJCIOMZ-WDSOQIARSA-N 0.000 description 1
- CZWIHKFGHICAJX-BPUTZDHNSA-N Trp-Glu-Glu Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 CZWIHKFGHICAJX-BPUTZDHNSA-N 0.000 description 1
- OJKVFAWXPGCJMF-BPUTZDHNSA-N Trp-Pro-Ser Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)N[C@@H](CO)C(=O)O OJKVFAWXPGCJMF-BPUTZDHNSA-N 0.000 description 1
- DTPWXZXGFAHEKL-NWLDYVSISA-N Trp-Thr-Glu Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O DTPWXZXGFAHEKL-NWLDYVSISA-N 0.000 description 1
- HTGJDTPQYFMKNC-VFAJRCTISA-N Trp-Thr-Leu Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C(O)=O)[C@@H](C)O)=CNC2=C1 HTGJDTPQYFMKNC-VFAJRCTISA-N 0.000 description 1
- 102000004243 Tubulin Human genes 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- MICSYKFECRFCTJ-IHRRRGAJSA-N Tyr-Arg-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O MICSYKFECRFCTJ-IHRRRGAJSA-N 0.000 description 1
- AYHSJESDFKREAR-KKUMJFAQSA-N Tyr-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AYHSJESDFKREAR-KKUMJFAQSA-N 0.000 description 1
- UXUFNBVCPAWACG-SIUGBPQLSA-N Tyr-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CC=C(C=C1)O)N UXUFNBVCPAWACG-SIUGBPQLSA-N 0.000 description 1
- FNWGDMZVYBVAGJ-XEGUGMAKSA-N Tyr-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC1=CC=C(C=C1)O)N FNWGDMZVYBVAGJ-XEGUGMAKSA-N 0.000 description 1
- BSCBBPKDVOZICB-KKUMJFAQSA-N Tyr-Leu-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O BSCBBPKDVOZICB-KKUMJFAQSA-N 0.000 description 1
- DWAMXBFJNZIHMC-KBPBESRZSA-N Tyr-Leu-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O DWAMXBFJNZIHMC-KBPBESRZSA-N 0.000 description 1
- ZOBLBMGJKVJVEV-BZSNNMDCSA-N Tyr-Lys-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)O)N)O ZOBLBMGJKVJVEV-BZSNNMDCSA-N 0.000 description 1
- PHKQVWWHRYUCJL-HJOGWXRNSA-N Tyr-Phe-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O PHKQVWWHRYUCJL-HJOGWXRNSA-N 0.000 description 1
- LVFZXRQQQDTBQH-IRIUXVKKSA-N Tyr-Thr-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O LVFZXRQQQDTBQH-IRIUXVKKSA-N 0.000 description 1
- MJUTYRIMFIICKL-JYJNAYRXSA-N Tyr-Val-Arg Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MJUTYRIMFIICKL-JYJNAYRXSA-N 0.000 description 1
- ASQFIHTXXMFENG-XPUUQOCRSA-N Val-Ala-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O ASQFIHTXXMFENG-XPUUQOCRSA-N 0.000 description 1
- JIODCDXKCJRMEH-NHCYSSNCSA-N Val-Arg-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N JIODCDXKCJRMEH-NHCYSSNCSA-N 0.000 description 1
- JLFKWDAZBRYCGX-ZKWXMUAHSA-N Val-Asn-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N JLFKWDAZBRYCGX-ZKWXMUAHSA-N 0.000 description 1
- QHDXUYOYTPWCSK-RCOVLWMOSA-N Val-Asp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)NCC(=O)O)N QHDXUYOYTPWCSK-RCOVLWMOSA-N 0.000 description 1
- YODDULVCGFQRFZ-ZKWXMUAHSA-N Val-Asp-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O YODDULVCGFQRFZ-ZKWXMUAHSA-N 0.000 description 1
- ZEVNVXYRZRIRCH-GVXVVHGQSA-N Val-Gln-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N ZEVNVXYRZRIRCH-GVXVVHGQSA-N 0.000 description 1
- AGKDVLSDNSTLFA-UMNHJUIQSA-N Val-Gln-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N AGKDVLSDNSTLFA-UMNHJUIQSA-N 0.000 description 1
- PMDOQZFYGWZSTK-LSJOCFKGSA-N Val-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)C(C)C PMDOQZFYGWZSTK-LSJOCFKGSA-N 0.000 description 1
- URIRWLJVWHYLET-ONGXEEELSA-N Val-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)C(C)C URIRWLJVWHYLET-ONGXEEELSA-N 0.000 description 1
- ZIGZPYJXIWLQFC-QTKMDUPCSA-N Val-His-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](C(C)C)N)O ZIGZPYJXIWLQFC-QTKMDUPCSA-N 0.000 description 1
- APEBUJBRGCMMHP-HJWJTTGWSA-N Val-Ile-Phe Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 APEBUJBRGCMMHP-HJWJTTGWSA-N 0.000 description 1
- JZWZACGUZVCQPS-RNJOBUHISA-N Val-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N JZWZACGUZVCQPS-RNJOBUHISA-N 0.000 description 1
- APQIVBCUIUDSMB-OSUNSFLBSA-N Val-Ile-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](C(C)C)N APQIVBCUIUDSMB-OSUNSFLBSA-N 0.000 description 1
- FEXILLGKGGTLRI-NHCYSSNCSA-N Val-Leu-Asn Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N FEXILLGKGGTLRI-NHCYSSNCSA-N 0.000 description 1
- BTWMICVCQLKKNR-DCAQKATOSA-N Val-Leu-Ser Chemical compound CC(C)[C@H]([NH3+])C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C([O-])=O BTWMICVCQLKKNR-DCAQKATOSA-N 0.000 description 1
- SYSWVVCYSXBVJG-RHYQMDGZSA-N Val-Leu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)N)O SYSWVVCYSXBVJG-RHYQMDGZSA-N 0.000 description 1
- ZRSZTKTVPNSUNA-IHRRRGAJSA-N Val-Lys-Leu Chemical compound CC(C)C[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)C(C)C)C(O)=O ZRSZTKTVPNSUNA-IHRRRGAJSA-N 0.000 description 1
- YMTOEGGOCHVGEH-IHRRRGAJSA-N Val-Lys-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O YMTOEGGOCHVGEH-IHRRRGAJSA-N 0.000 description 1
- MHHAWNPHDLCPLF-ULQDDVLXSA-N Val-Phe-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC1=CC=CC=C1 MHHAWNPHDLCPLF-ULQDDVLXSA-N 0.000 description 1
- VHIZXDZMTDVFGX-DCAQKATOSA-N Val-Ser-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N VHIZXDZMTDVFGX-DCAQKATOSA-N 0.000 description 1
- CEKSLIVSNNGOKH-KZVJFYERSA-N Val-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](C(C)C)N)O CEKSLIVSNNGOKH-KZVJFYERSA-N 0.000 description 1
- GVNLOVJNNDZUHS-RHYQMDGZSA-N Val-Thr-Lys Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(O)=O GVNLOVJNNDZUHS-RHYQMDGZSA-N 0.000 description 1
- PDDJTOSAVNRJRH-UNQGMJICSA-N Val-Thr-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](C(C)C)N)O PDDJTOSAVNRJRH-UNQGMJICSA-N 0.000 description 1
- XNLUVJPMPAZHCY-JYJNAYRXSA-N Val-Val-Phe Chemical compound CC(C)[C@H]([NH3+])C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C([O-])=O)CC1=CC=CC=C1 XNLUVJPMPAZHCY-JYJNAYRXSA-N 0.000 description 1
- 241001416177 Vicugna pacos Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- UBORTCNDUKBEOP-HAVMAKPUSA-N Xanthosine Natural products O[C@@H]1[C@H](O)[C@H](CO)O[C@H]1N1C(NC(=O)NC2=O)=C2N=C1 UBORTCNDUKBEOP-HAVMAKPUSA-N 0.000 description 1
- HCAJCMUKLZSPFT-KWXKLSQISA-N [3-(dimethylamino)-2-[(9z,12z)-octadeca-9,12-dienoyl]oxypropyl] (9z,12z)-octadeca-9,12-dienoate Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(=O)OCC(CN(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC HCAJCMUKLZSPFT-KWXKLSQISA-N 0.000 description 1
- HMNZFMSWFCAGGW-XPWSMXQVSA-N [3-[hydroxy(2-hydroxyethoxy)phosphoryl]oxy-2-[(e)-octadec-9-enoyl]oxypropyl] (e)-octadec-9-enoate Chemical compound CCCCCCCC\C=C\CCCCCCCC(=O)OCC(COP(O)(=O)OCCO)OC(=O)CCCCCCC\C=C\CCCCCCCC HMNZFMSWFCAGGW-XPWSMXQVSA-N 0.000 description 1
- BAQODHZVQKWPMS-UHFFFAOYSA-N [Si](O)(O)(O)O.[P].[Ca] Chemical compound [Si](O)(O)(O)O.[P].[Ca] BAQODHZVQKWPMS-UHFFFAOYSA-N 0.000 description 1
- YCZSHICNESTART-ZOQUXTDFSA-N [[(2r,3r,4r,5r)-5-(4-amino-2-oxopyrimidin-1-yl)-3-hydroxy-4-methoxyoxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound CO[C@@H]1[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O[C@H]1N1C(=O)N=C(N)C=C1 YCZSHICNESTART-ZOQUXTDFSA-N 0.000 description 1
- WNVZQYHBHSLUHJ-XVFCMESISA-N [[(2r,3s,4r,5r)-4-amino-5-(4-amino-2-oxopyrimidin-1-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound N[C@@H]1[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O[C@H]1N1C(=O)N=C(N)C=C1 WNVZQYHBHSLUHJ-XVFCMESISA-N 0.000 description 1
- NCKFQXVRKKNRBB-SHUUEZRQSA-N [[(2r,3s,4r,5r)-5-(3,5-dioxo-1,2,4-triazin-2-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound O[C@@H]1[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O[C@H]1N1C(=O)NC(=O)C=N1 NCKFQXVRKKNRBB-SHUUEZRQSA-N 0.000 description 1
- YIJVOACVHQZMKI-JXOAFFINSA-N [[(2r,3s,4r,5r)-5-(4-amino-5-methyl-2-oxopyrimidin-1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound O=C1N=C(N)C(C)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 YIJVOACVHQZMKI-JXOAFFINSA-N 0.000 description 1
- PQISXOFEOCLOCT-UUOKFMHZSA-N [[(2r,3s,4r,5r)-5-(6-amino-8-azidopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound [N-]=[N+]=NC1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O PQISXOFEOCLOCT-UUOKFMHZSA-N 0.000 description 1
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- UDMBCSSLTHHNCD-KQYNXXCUSA-N adenosine 5'-monophosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H]1O UDMBCSSLTHHNCD-KQYNXXCUSA-N 0.000 description 1
- 229960001456 adenosine triphosphate Drugs 0.000 description 1
- WNLRTRBMVRJNCN-UHFFFAOYSA-L adipate(2-) Chemical compound [O-]C(=O)CCCCC([O-])=O WNLRTRBMVRJNCN-UHFFFAOYSA-L 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 1
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 108010070783 alanyltyrosine Proteins 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001447 alkali salts Chemical class 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 125000003302 alkenyloxy group Chemical group 0.000 description 1
- 150000001350 alkyl halides Chemical class 0.000 description 1
- 125000001118 alkylidene group Chemical group 0.000 description 1
- 208000030961 allergic reaction Diseases 0.000 description 1
- AEMOLEFTQBMNLQ-BKBMJHBISA-M alpha-D-galacturonate Chemical compound O[C@H]1O[C@H](C([O-])=O)[C@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-BKBMJHBISA-M 0.000 description 1
- AWUCVROLDVIAJX-UHFFFAOYSA-N alpha-glycerophosphate Natural products OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000009435 amidation Effects 0.000 description 1
- 238000007112 amidation reaction Methods 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- SWLVFNYSXGMGBS-UHFFFAOYSA-N ammonium bromide Chemical compound [NH4+].[Br-] SWLVFNYSXGMGBS-UHFFFAOYSA-N 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 108010008355 arginyl-glutamine Proteins 0.000 description 1
- 229940072107 ascorbate Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 229940009098 aspartate Drugs 0.000 description 1
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- 108010028263 bacteriophage T3 RNA polymerase Proteins 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 229940077388 benzenesulfonate Drugs 0.000 description 1
- SRSXLGNVWSONIS-UHFFFAOYSA-M benzenesulfonate Chemical compound [O-]S(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-M 0.000 description 1
- 229940050390 benzoate Drugs 0.000 description 1
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 229910000085 borane Inorganic materials 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical class OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 210000000621 bronchi Anatomy 0.000 description 1
- 150000007516 brønsted-lowry acids Chemical class 0.000 description 1
- 150000007528 brønsted-lowry bases Chemical class 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229960000846 camphor Drugs 0.000 description 1
- 229930008380 camphor Natural products 0.000 description 1
- 229950005953 camsilate Drugs 0.000 description 1
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000011203 carbon fibre reinforced carbon Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 150000007942 carboxylates Chemical class 0.000 description 1
- 241001233037 catfish Species 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 150000005829 chemical entities Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- XTEGARKTQYYJKE-UHFFFAOYSA-N chloric acid Chemical compound OCl(=O)=O XTEGARKTQYYJKE-UHFFFAOYSA-N 0.000 description 1
- 229940005991 chloric acid Drugs 0.000 description 1
- 229940099352 cholate Drugs 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- BALGDZWGNCXXES-UHFFFAOYSA-N cyclopentane;propanoic acid Chemical compound CCC(O)=O.C1CCCC1 BALGDZWGNCXXES-UHFFFAOYSA-N 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 210000004292 cytoskeleton Anatomy 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 125000002704 decyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000011036 discontinuous diafiltration Methods 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- VFNGKCDDZUSWLR-UHFFFAOYSA-L disulfate(2-) Chemical compound [O-]S(=O)(=O)OS([O-])(=O)=O VFNGKCDDZUSWLR-UHFFFAOYSA-L 0.000 description 1
- 125000003438 dodecyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 229940043264 dodecyl sulfate Drugs 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 210000001755 duct epithelial cell Anatomy 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 210000003038 endothelium Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229950007655 esilate Drugs 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000012065 filter cake Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 235000019688 fish Nutrition 0.000 description 1
- 229960002949 fluorouracil Drugs 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 150000004675 formic acid derivatives Chemical class 0.000 description 1
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 description 1
- 238000004508 fractional distillation Methods 0.000 description 1
- VZCYOOQTPOCHFL-OWOJBTEDSA-L fumarate(2-) Chemical compound [O-]C(=O)\C=C\C([O-])=O VZCYOOQTPOCHFL-OWOJBTEDSA-L 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 150000002243 furanoses Chemical group 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 1
- 238000003209 gene knockout Methods 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 108060003196 globin Proteins 0.000 description 1
- 102000018146 globin Human genes 0.000 description 1
- 229960001731 gluceptate Drugs 0.000 description 1
- KWMLJOLKUYYJFJ-VFUOTHLCSA-N glucoheptonic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O)C(O)=O KWMLJOLKUYYJFJ-VFUOTHLCSA-N 0.000 description 1
- 229940050410 gluconate Drugs 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 108010079547 glutamylmethionine Proteins 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- 108010038983 glycyl-histidyl-lysine Proteins 0.000 description 1
- 108010077435 glycyl-phenylalanyl-glycine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- ZRALSGWEFCBTJO-UHFFFAOYSA-N guanidine group Chemical group NC(=N)N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 1
- 150000002357 guanidines Chemical class 0.000 description 1
- 150000004820 halides Chemical class 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- MNWFXJYAOYHMED-UHFFFAOYSA-M heptanoate Chemical compound CCCCCCC([O-])=O MNWFXJYAOYHMED-UHFFFAOYSA-M 0.000 description 1
- IPCSVZSSVZVIGE-UHFFFAOYSA-M hexadecanoate Chemical compound CCCCCCCCCCCCCCCC([O-])=O IPCSVZSSVZVIGE-UHFFFAOYSA-M 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 125000001183 hydrocarbyl group Chemical group 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 238000013115 immunohistochemical detection Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000006202 intradermal dosage form Substances 0.000 description 1
- 238000007913 intrathecal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- SUMDYPCJJOFFON-UHFFFAOYSA-N isethionic acid Chemical class OCCS(O)(=O)=O SUMDYPCJJOFFON-UHFFFAOYSA-N 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 108010053037 kyotorphin Proteins 0.000 description 1
- 230000006651 lactation Effects 0.000 description 1
- 229940099584 lactobionate Drugs 0.000 description 1
- JYTUSYBCFIZPBE-AMTLMPIISA-M lactobionate Chemical compound [O-]C(=O)[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O JYTUSYBCFIZPBE-AMTLMPIISA-M 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 108010077158 leucinyl-arginyl-tryptophan Proteins 0.000 description 1
- 108010083708 leucyl-aspartyl-valine Proteins 0.000 description 1
- 108010044056 leucyl-phenylalanine Proteins 0.000 description 1
- 108010000761 leucylarginine Proteins 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- SMEROWZSTRWXGI-HVATVPOCSA-N lithocholic acid 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](CCC(O)=O)C)[C@@]2(C)CC1 SMEROWZSTRWXGI-HVATVPOCSA-N 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 230000002132 lysosomal effect Effects 0.000 description 1
- 108010003700 lysyl aspartic acid Proteins 0.000 description 1
- 108010054155 lysyllysine Proteins 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 229920001427 mPEG Polymers 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 229940049920 malate Drugs 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-L malate(2-) Chemical compound [O-]C(=O)C(O)CC([O-])=O BJEPYKJPYRNKOW-UHFFFAOYSA-L 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 108010068488 methionylphenylalanine Proteins 0.000 description 1
- OYQVKYFWJZYYOA-UHFFFAOYSA-N methylamino dihydrogen phosphate Chemical compound CNOP(O)(O)=O OYQVKYFWJZYYOA-UHFFFAOYSA-N 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- XBCXJKGHPABGSD-UHFFFAOYSA-N methyluracil Natural products CN1C=CC(=O)NC1=O XBCXJKGHPABGSD-UHFFFAOYSA-N 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 210000000107 myocyte Anatomy 0.000 description 1
- NFQBIAXADRDUGK-KWXKLSQISA-N n,n-dimethyl-2,3-bis[(9z,12z)-octadeca-9,12-dienoxy]propan-1-amine Chemical compound CCCCC\C=C/C\C=C/CCCCCCCCOCC(CN(C)C)OCCCCCCCC\C=C/C\C=C/CCCCC NFQBIAXADRDUGK-KWXKLSQISA-N 0.000 description 1
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000002088 nanocapsule Substances 0.000 description 1
- KVBGVZZKJNLNJU-UHFFFAOYSA-M naphthalene-2-sulfonate Chemical compound C1=CC=CC2=CC(S(=O)(=O)[O-])=CC=C21 KVBGVZZKJNLNJU-UHFFFAOYSA-M 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 229940049964 oleate Drugs 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-M oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC([O-])=O ZQPPMHVWECSIRJ-KTKRTIGZSA-M 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 150000003901 oxalic acid esters Chemical class 0.000 description 1
- 235000010603 pastilles Nutrition 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- JRKICGRDRMAZLK-UHFFFAOYSA-L peroxydisulfate Chemical compound [O-]S(=O)(=O)OOS([O-])(=O)=O JRKICGRDRMAZLK-UHFFFAOYSA-L 0.000 description 1
- 108010084572 phenylalanyl-valine Proteins 0.000 description 1
- 108010018625 phenylalanylarginine Proteins 0.000 description 1
- 150000004713 phosphodiesters Chemical group 0.000 description 1
- UEZVMMHDMIWARA-UHFFFAOYSA-M phosphonate Chemical compound [O-]P(=O)=O UEZVMMHDMIWARA-UHFFFAOYSA-M 0.000 description 1
- 229940075930 picrate Drugs 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-M picrate anion Chemical compound [O-]C1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-M 0.000 description 1
- IUGYQRQAERSCNH-UHFFFAOYSA-M pivalate Chemical compound CC(C)(C)C([O-])=O IUGYQRQAERSCNH-UHFFFAOYSA-M 0.000 description 1
- 229950010765 pivalate Drugs 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 229920001515 polyalkylene glycol Polymers 0.000 description 1
- 229920000151 polyglycol Polymers 0.000 description 1
- 239000010695 polyglycol Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- 108010090894 prolylleucine Proteins 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 239000003725 proteoliposome Substances 0.000 description 1
- 229950010131 puromycin Drugs 0.000 description 1
- PBMFSQRYOILNGV-UHFFFAOYSA-N pyridazine Chemical group C1=CC=NN=C1 PBMFSQRYOILNGV-UHFFFAOYSA-N 0.000 description 1
- 125000001453 quaternary ammonium group Chemical group 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000003161 ribonuclease inhibitor Substances 0.000 description 1
- 239000002336 ribonucleotide Substances 0.000 description 1
- 125000002652 ribonucleotide group Chemical group 0.000 description 1
- 108020004418 ribosomal RNA Proteins 0.000 description 1
- 210000003705 ribosome Anatomy 0.000 description 1
- DWRXFEITVBNRMK-JXOAFFINSA-N ribothymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 DWRXFEITVBNRMK-JXOAFFINSA-N 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 210000003079 salivary gland Anatomy 0.000 description 1
- 235000019515 salmon Nutrition 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 238000005549 size reduction Methods 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- AWUCVROLDVIAJX-GSVOUGTGSA-N sn-glycerol 3-phosphate Chemical compound OC[C@@H](O)COP(O)(O)=O AWUCVROLDVIAJX-GSVOUGTGSA-N 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 239000001384 succinic acid Substances 0.000 description 1
- 150000003460 sulfonic acids Chemical class 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000013077 target material Substances 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 229940095064 tartrate Drugs 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 150000003580 thiophosphoric acid esters Chemical class 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 108010071097 threonyl-lysyl-proline Proteins 0.000 description 1
- 108010072986 threonyl-seryl-lysine Proteins 0.000 description 1
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 229950004288 tosilate Drugs 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 108091005703 transmembrane proteins Proteins 0.000 description 1
- 102000035160 transmembrane proteins Human genes 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
- 108010036387 trimethionine Proteins 0.000 description 1
- 108010044292 tryptophyltyrosine Proteins 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- ZDPHROOEEOARMN-UHFFFAOYSA-N undecanoic acid Chemical compound CCCCCCCCCCC(O)=O ZDPHROOEEOARMN-UHFFFAOYSA-N 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
- 229940070710 valerate Drugs 0.000 description 1
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N valeric acid Chemical compound CCCCC(O)=O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 210000004885 white matter Anatomy 0.000 description 1
- WCNMEQDMUYVWMJ-JPZHCBQBSA-N wybutoxosine Chemical compound C1=NC=2C(=O)N3C(CC([C@H](NC(=O)OC)C(=O)OC)OO)=C(C)N=C3N(C)C=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O WCNMEQDMUYVWMJ-JPZHCBQBSA-N 0.000 description 1
- UBORTCNDUKBEOP-UUOKFMHZSA-N xanthosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(NC(=O)NC2=O)=C2N=C1 UBORTCNDUKBEOP-UUOKFMHZSA-N 0.000 description 1
Classifications
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/554—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- 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
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6905—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion
- A61K47/6911—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion the form being a liposome
-
- 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
- A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
-
- 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
- A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
- A61K48/0025—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid
- A61K48/0033—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid the non-active part being non-polymeric
-
- 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
- A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
- A61K48/0025—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid
- A61K48/0041—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid the non-active part being polymeric
-
- 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
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
-
- 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
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
- A61K48/0066—Manipulation of the nucleic acid to modify its expression pattern, e.g. enhance its duration of expression, achieved by the presence of particular introns in the delivered nucleic 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
- A61K48/0075—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the delivery route, e.g. oral, subcutaneous
-
- 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
- A61K9/007—Pulmonary tract; Aromatherapy
- A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
- A61K9/0078—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy for inhalation via a nebulizer such as a jet nebulizer, ultrasonic nebulizer, e.g. in the form of aqueous drug solutions or dispersions
-
- 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
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
- A61K9/1272—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids
-
- 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
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/12—Mucolytics
Abstract
本发明尤其提供了配制含核酸的纳米颗粒的组合物和方法,所述纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质。在一些实施例中,本发明提供了其中所述脂质纳米颗粒进一步包含辅助脂质和PEG改性脂质的组合物和方法。所得制剂包含高核酸囊封百分比。
Description
相关申请
本申请要求2016年11月10日提交的美国临时申请第62/420,421号、2016年11月10日提交的美国临时申请第62/420,428号、2016年11月11日提交的美国临时申请第62/421,021号、2016年11月11日提交的美国临时申请第62/421,007号、2017年2月27日提交的美国临时申请第62/464,327号和2017年2月27日提交的美国临时申请第62/464,330号的优先权,其公开内容以引用的方式并入本文中。
序列表
本说明书参考了序列表(在2017年11月10日以电子方式作为.txt文件提交,命名为“MRT-1247WO_SL”)。.txt文件于2017年11月10日生成,并且大小为57,411字节。序列表的全部内容以引用的方式并入本文。
背景技术
基于核酸的技术对于各种治疗应用越来越重要,所述治疗应用包括但不限于信使RNA疗法。递送核酸的努力包括产生被配制成保护核酸在体内递送时免受降解的组合物。核酸的一种类型的递送媒剂为脂质纳米颗粒。将脂质纳米颗粒成功地用作递送媒剂所考虑的重要参数包括脂质纳米颗粒形成、脂质组分的物理特性、核酸囊封效率、体内核酸释放潜力和脂质纳米颗粒毒性。
发明内容
本发明提供了一种独特的解决方案,其中基于固醇的阳离子脂质、辅助脂质和PEG改性脂质形成RNA的脂质纳米颗粒制剂。本发明的三种脂质组分系统在配制后显示出高RNA囊封效率和成功的有效体内递送,特别是肺部递送。此类制剂系统提供更低脂质负载(与其它常规的四脂质组分系统相比)和更高耐受性/更低毒性的额外优点,因为阳离子脂质(胆固醇衍生脂质)的代谢产物是胆固醇。
本发明部分地基于以下出人意料的发现:基于基于固醇的阳离子脂质的三脂质组分系统在体内,特别是在肺中递送mRNA并产生编码的蛋白质或肽方面出乎意料地有效。实际上,在本发明之前,阳离子脂质已被广泛地作为用于囊封核酸(包括mRNA)以用于体内递送的脂质体的重要组分而研究。由于mRNA的独特脆性长结构和复杂的体内翻译过程,脂质体中使用的阳离子脂质通常起两种作用。首先,阳离子脂质促进在囊封、循环和内吞期间与带负电mRNA的相互作用,从而捕获和保护mRNA。然后,一旦在细胞溶质内部,阳离子脂质便需要能够释放mRNA,以使得mRNA可以被翻译以产生编码的蛋白质或肽。一些阳离子脂质(特别是称为可滴定阳离子脂质)在递送mRNA中尤其有效。出人意料地,本发明人发现,包含本文所述的基于固醇的阳离子脂质的脂质体可具有更高的mRNA囊封百分比,并且可在体内递送各种mRNA中更有效。具体来说,包含本文所述的基于固醇的阳离子脂质的脂质体对于肺部递送mRNA来说可能是极其有效的,并且对于经由喷雾递送mRNA来说出人意料地成功。因此,本发明人已表明,三脂质组分系统可以独特地用于递送mRNA以用于在体内,特别是在肺部系统中高效和持续生产蛋白质或肽(例如,治疗蛋白)。因此,本发明允许改进的mRNA疗法,其可显著减少mRNA和相关的脂质的所需量、施用频率和可能的副作用,从而为各种疾病提供更安全、更有效和患者友好的mRNA疗法。
在一个方面,本发明提供了在体内递送核酸的方法,其包含通过肺部递送向需要递送的个体施用包含核酸和囊封核酸的脂质纳米颗粒的组合物,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质。
在另一方面,本发明提供了在体内递送核酸的方法,其包括通过肺部递送向需要递送的个体施用组合物,其包含核酸,其中核酸编码囊性纤维化跨膜传导调节(CFTR)蛋白,和囊封所述核酸的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质。
在一些实施例中,肺部递送包含喷雾。在一些实施例中,所述个体是需要递送的个体。
在一些实施例中,脂质纳米颗粒具有至少70%(例如至少75%、至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%)的核酸囊封百分比。
在一些实施例中,三种不同的脂质组分包含辅助脂质和PEG改性脂质中的一种或两种。
在一些实施例中,基于固醇的阳离子脂质具有根据式(A)的结构,
B-L1-S(式A)或其质子化形式,其中B是碱性官能团,其中质子化形式具有不超过约8.0的pKa;L1是任选经取代的连接基团,其为C1-C20亚烷基或2至20元亚杂烷基;且S是固醇。
在一些实施例中,B是任选经取代的5元或6元含氮杂芳基。在一些实施例中,B是选自吡咯基、咪唑基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。在一些实施例中,B是选自吡咯基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。在一些实施例中,B是选自吡咯基、吡唑基、三唑基、四唑基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
在一些实施例中,L1是任选经取代的连接基团,其为C1-C20亚烷基。在一些实施例中,L1是任选经取代的连接基团,其为2至20元亚杂烷基。在一些实施例中,L1是非肽的2至20元亚杂烷基。在一些实施例中,L1包含为酯基、酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。在一些实施例中,L1包含为酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。在一些实施例中,L1包含作为酰胺基、碳酸酯基或脲基的部分。在一些实施例中,L1为-X1-C(X3)-X2、-(C1-C19亚烷基)-X1-C(X3)-X2、-X1-C(X3)-X2(C1-C19亚烷基)-、-(C1-C19亚烷基)-X1-、-X1-(C1-C19亚烷基)-,其中每个X1和X2独立地为共价键、-O-、-S-或-NH-;X3独立地为=O、=S或=NH;且其中所述C1-C19亚烷基任选经取代。在一些实施例中,L1不包含具有结构-N(R')2的取代基,或其带正电形式,其中每个R'独立地为氢或任选经取代的C1-C20烷基。
在一些实施例中,S是动物固醇,或其氧化或还原形式。在一些实施例中,S是植物固醇,或其氧化或还原形式。在一些实施例中,S是合成固醇,或其氧化或还原形式。在一些实施例中,S是选自胆固醇、胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。在一些实施例中,S是选自胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。在一些实施例中,S是选自以下的固醇:
其中R是任选经取代的C1-C20烷基。在一些实施例中,S是选自以下的固醇:
其中R是任选经取代的C1-C20烷基。
在一些实施例中,基于固醇的阳离子脂质包含咪唑胆固醇酯(ICE)。在一些实施例中,基于固醇的阳离子脂质不包含咪唑胆固醇酯(ICE)。
在一些实施例中,核酸选自DNA、siRNA、微RNA和/或mRNA。在一些实施例中,核酸是编码蛋白质或肽的mRNA。在一些实施例中,编码蛋白质或肽的mRNA是密码子优化的。在一些实施例中,mRNA包含一种或多种经修饰的核苷酸。在一些实施例中,mRNA包含对所述mRNA的5'非翻译区的修饰。在一些实施例中,对5'非翻译区的所述修饰包含将Cap1结构包括在内。在一些实施例中,mRNA包含对所述mRNA的3'非翻译区的修饰。在一些实施例中,3'非翻译区的所述修饰包含将poly A尾包括在内。
在一些实施例中,脂质纳米颗粒具有小于约100nm、95nm、90nm、85nm、80nm、75nm、70nm、65nm、60nm、55nm、50nm、45nm或40nm的尺寸。
在另一方面,本发明提供配制用于喷雾的组合物,其包含核酸和囊封核酸的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质,且另外,其中脂质纳米颗粒具有至少70%(例如至少75%、至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%)的核酸囊封百分比。
在另一方面,本发明提供组合物,其包含核酸和囊封核酸的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质,且另外,其中脂质纳米颗粒具有至少70%(例如至少75%、至少80%、至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%)的核酸囊封百分比。
在一些实施例中,脂质纳米颗粒具有至少85%的核酸囊封百分比。在一些实施例中,脂质纳米颗粒具有至少90%的核酸囊封百分比。
在另一方面,本发明提供组合物,其包含编码囊性纤维化跨膜传导调节(CFTR)蛋白的mRNA和囊封mRNA的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同脂质组分是基于固醇的阳离子脂质,且另外,其中脂质纳米颗粒具有至少80%的mRNA囊封百分比。
在一些实施例中,脂质纳米颗粒具有至少85%或至少90%的mRNA囊封百分比。
在另一方面,本发明提供组合物,其包含核酸和囊封核酸的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质,且另外,其中基于固醇的阳离子脂质占总脂质的不超过70%(例如不超过68%、不超过67%、不超过66%、不超过65%、不超过60%、不超过55%、不超过50%、不超过45%或不超过40%)。
在另一方面,本发明提供组合物,其包含编码囊性纤维化跨膜传导调节(CFTR)蛋白的mRNA和囊封mRNA的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质,且另外,其中基于固醇的阳离子脂质占总脂质的不超过70%(例如不超过68%、不超过67%、不超过66%、不超过65%、不超过60%、不超过55%、不超过50%、不超过45%或不超过40%)。
在一些实施例中,基于固醇的阳离子脂质占总脂质的不超过65%。在一些实施例中,基于固醇的阳离子脂质占总脂质的不超过60%。在一些实施例中,三种不同的脂质组分包含辅助脂质和PEG改性脂质中的一种或两种。
在一些实施例中,基于固醇的阳离子脂质具有根据式(A)的结构,
B-L1-S(式A)或其质子化形式,其中B是碱性官能团,其中质子化形式具有不超过约8.0的pKa;L1是任选经取代的连接基团,其为C1-C20亚烷基或2至20元亚杂烷基;且S是固醇。
在一些实施例中,B是任选经取代的5元或6元含氮杂芳基。在一些实施例中,B是选自吡咯基、咪唑基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。在一些实施例中,B是选自吡咯基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。在一些实施例中,B是选自吡咯基、吡唑基、三唑基、四唑基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
在一些实施例中,L1是任选经取代的连接基团,其为C1-C20亚烷基。在一些实施例中,L1是任选经取代的连接基团,其为2至20元亚杂烷基。在一些实施例中,L1是非肽的2至20元亚杂烷基。在一些实施例中,L1包含为酯基、酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。在一些实施例中,L1包含为酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。在一些实施例中,L1包含作为酰胺基、碳酸酯基或脲基的部分。在一些实施例中,L1为-X1-C(X3)-X2、-(C1-C19亚烷基)-X1-C(X3)-X2、-X1-C(X3)-X2(C1-C19亚烷基)-、-(C1-C19亚烷基)-X1-、-X1-(C1-C19亚烷基)-,其中每个X1和X2独立地为共价键、-O-、-S-或-NH-;X3独立地为=O、=S或=NH;且其中所述C1-C19亚烷基任选经取代。在一些实施例中,L1不包含具有结构-N(R')2的取代基,或其带正电形式,其中每个R'独立地为氢或任选经取代的C1-C20烷基。
在一些实施例中,S是动物固醇,或其氧化或还原形式。在一些实施例中,S是植物固醇,或其氧化或还原形式。在一些实施例中,S是合成固醇,或其氧化或还原形式。在一些实施例中,S是选自胆固醇、胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。在一些实施例中,S是选自胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。在一些实施例中,S是选自以下的固醇:
其中R是任选经取代的C1-C20烷基。在一些实施例中,S是选自以下的固醇:
其中R是任选经取代的C1-C20烷基。
在一些实施例中,基于固醇的阳离子脂质包含咪唑胆固醇酯(ICE)。在一些实施例中,基于固醇的阳离子脂质不包含咪唑胆固醇酯(ICE)。
在一些实施例中,核酸选自DNA、siRNA、微RNA和/或mRNA。在一些实施例中,核酸是编码蛋白质或肽的mRNA。在一些实施例中,编码蛋白质或肽的mRNA是密码子优化的。在一些实施例中,mRNA包含一种或多种经修饰的核苷酸。在一些实施例中,mRNA包含对所述mRNA的5'非翻译区的修饰。在一些实施例中,对5'非翻译区的所述修饰包含将Cap1结构包括在内。在一些实施例中,mRNA包含对所述mRNA的3'非翻译区的修饰。在一些实施例中,3'非翻译区的所述修饰包含将poly A尾包括在内。
在一些实施例中,脂质纳米颗粒具有小于约100nm、95nm、90nm、85nm、80nm、75nm、70nm、65nm、60nm、55nm、50nm、45nm或40nm的尺寸。
在一些实施例中,编码CFTR蛋白的mRNA包含SEQ ID NO:1。在一些实施例中,编码CFTR蛋白的mRNA包含与SEQ ID NO:1至少70%、75%、80%、85%、90%或95%一致的多核苷酸序列。
在一些实施例中,脂质纳米颗粒通过将mRNA溶液和脂质溶液混合到20%乙醇中而形成。在一些实施例中,脂质纳米颗粒通过切向流过滤进一步纯化。
在另一方面,本发明提供了在体内递送mRNA的方法,其包含向个体施用组合物。在一些实施例中,所述个体是需要递送的个体。在一些实施例中,组合物是静脉内施用的。在一些实施例中,组合物通过肺部递送施用。在一些实施例中,肺部递送包含喷雾。
在本申请中,除非另有说明,否则使用“或”意指“和/或”。如本公开中所使用,术语“包含(comprise)”和所述术语的变化形式,例如“包含(comprising)”和“包含(comprises)”并不打算排除其它添加剂、组分、整数或步骤。如本申请中所使用,术语“约”和“大约”用作等效物。这两个术语均旨在涵盖相关领域的普通技术人员所理解的任何正常波动。
在下述详细描述、附图和权利要求书中,本发明的其它特征、目的和优点是显而易见的。然而,应理解,详细描述、附图和权利要求书虽然指出了本发明的实施例,但仅以说明性方式而非限制性方式给出。在本发明的范围内的各种变化和修改对于所属领域的技术人员将变得显而易见。
附图说明
附图仅用于说明性目的,而不是限制。
图1描绘在用囊封密码子优化的hCFTR mRNA的基于ICE的LNP处理之后24小时,小鼠肺中的人囊性纤维化跨膜传导调节蛋白(hCFTR)的示例性免疫组织化学检测,B=分别在10×和20×放大率下的未处理的小鼠肺。C、D=分别在10×和20×放大率下的CO-hCFTRmRNA ICE LNP处理的小鼠肺。
图2描绘在用囊封密码子优化的hCFTR mRNA的基于ICE的LNP处理之后24小时,小鼠肺中的人类CFTR蛋白的示例性免疫组织化学检测。A、B=分别在10×和20×放大率下的未处理的小鼠肺。C、D=分别在10×和20×放大率下的CO-hCFTR mRNA ICE LNP处理的小鼠肺。
图3描绘在用囊封密码子优化的hCFTR mRNA的雾化的基于ICE的脂质纳米颗粒(LNP)(5%PEG)处理之后24小时,大鼠肺中的人囊性纤维化跨膜传导调节蛋白(hCFTR)的示例性免疫组织化学检测。
图4描绘在经由气管内施用用囊封密码子优化的hCFTR mRNA的基于ICE的LNP(5%PEG)处理之后24小时,大鼠肺中的hCFTR蛋白的示例性免疫组织化学检测。
图5描绘在用囊封密码子优化的(CO)hCFTR mRNA的雾化的基于ICE的LNP(3%PEG)处理后24小时,大鼠肺中的人CFTR蛋白的示例性免疫组织化学检测。
图6描绘在经由气管内(微型喷雾器)施用用囊封CO-hCFTR mRNA的基于ICE的LNP(3%PEG)处理后24小时,大鼠肺中的人CFTR蛋白的示例性免疫组织化学检测。
图7描绘在用囊封CO-hCFTR mRNA的雾化的基于ICE的LNP(N/P=2)处理后24小时,CFTR KO(基因敲除)小鼠肺中的人CFTR蛋白的示例性免疫组织化学检测。
图8描绘在用囊封CO-hCFTR mRNA的雾化的基于ICE的LNP(N/P=4)处理后24小时,CFTR KO小鼠肺中的人CFTR蛋白的示例性免疫组织化学检测。
图9描绘在用囊封各种mRNA构筑体,包括CFTR(CO-hCFTR mRNA)、STOP(不能翻译为蛋白质的无义突变CO-hCFTR mRNA)和FFL(萤火虫荧光素酶mRNA)的雾化的基于ICE的LNP处理后24小时,野生型小鼠肺中的人CFTR蛋白的示例性免疫组织化学检测。
图10描绘在单次暴露于囊封CO-hCFTR mRNA的雾化的基于ICE的LNP后的各个时间点,野生型小鼠肺中的人CFTR蛋白的示例性免疫组织化学检测。
图11描绘冷冻肺切片中CO-hCFTR mRNA的拷贝/微克总RNA的比率的示例图,所述冷冻肺切片来自用对照(缓冲液,空ICE LNP)或载有CO-hCFTR mRNA的ICE LNP处理的大鼠。
图12描绘用对照(缓冲液,空ICE LNP)或载有CO-hCFTR mRNA的ICE LNP处理的大鼠的冷冻肺切片中CO-hCFTR的拷贝相比于CFTR mRNA的内源性水平的倍数增加的示例图。
图13描绘在单次暴露于各种剂量的囊封CO-hCFTR mRNA的雾化的基于ICE的LNP之后,大鼠肺中的人CFTR蛋白的示例性免疫组织化学检测。
图14A和14B描绘相对于对照,用支链PEI(图14A)或ICE(图14B)配制的各种剂量的hCFTR mRNA对大鼠的呼吸速率的影响的示例图。
图15描绘显示通过hCFTR mRNA诱导的氯离子通道活性的剂量反应的示例图。
具体实施方式
定义
为了使本发明更容易理解,首先在下文定义了某些术语。下述术语和其它术语的另外定义阐述在整个本说明书中。本文引用的出版物和其它参考材料描述本发明的背景且提供关于其实践的其它细节,以引用的方式并入本文中。
烷基:如本文中所用,“烷基”是指具有1至15个碳原子的直链或支链饱和烃基的基团(“C1-15烷基”)。在一些实施例中,烷基具有1至3个碳原子(“C1-3烷基”)。C1-3烷基的实例包括甲基(C1)、乙基(C2)、正丙基(C3)和异丙基(C3)。在一些实施例中,烷基具有8至12个碳原子(“C8-12烷基”)。C8-12烷基的实例包括但不限于正辛基(C8)、正壬基(C9)、正癸基(C10)、正十一烷基(C11)、正十二烷基(C12)等。前缀“n-”(正)是指未分支的烷基。例如,n-C8烷基是指-(CH2)7CH3,n-C10烷基是指-(CH2)9CH3等。
氨基酸:如本文所用,术语“氨基酸”在其最广泛的意义上是指可并入多肽链内的任何化合物和/或物质。在一些实施例中,氨基酸具有一般结构H2N-C(H)(R)-COHO。在一些实施例中,氨基酸是天然存在的氨基酸。在一些实施例中,氨基酸是合成氨基酸;在一些实施例中,氨基酸是d-氨基酸;在一些实施例中,氨基酸是l-氨基酸。“标准氨基酸”是指天然存在的肽中常见的二十种标准l-氨基酸中的任一种。“非标准氨基酸”是指除标准氨基酸外的任何氨基酸,无论其为以合成方式制备的或获自天然来源的。如本文所用,“合成氨基酸”涵盖化学修饰的氨基酸,包括但不限于盐、氨基酸衍生物(例如酰胺)和/或取代。氨基酸,包括肽中的羧基和/或氨基末端氨基酸,可通过甲基化、酰胺化、乙酰化、保护基和/或用其它化学基团取代来修饰,所述其它化学基团可改变肽的循环半衰期,而不会不利地影响其活性。氨基酸可参与二硫键。氨基酸可包含一个或翻译后修饰,例如与一个或多个化学实体(例如,甲基、乙酸基、乙酰基、磷酸基、甲酰基部分、类异戊二烯基、硫酸基、聚乙二醇部分、脂质部分、碳水化合物部分、生物素部分等)结合。术语“氨基酸”可与“氨基酸残基”互换使用,并且可指游离氨基酸和/或肽的氨基酸残基。从使用所述术语的上下文中显而易见它是指游离氨基酸还是肽的残基。
动物:如本文所用,术语“动物”是指动物界的任何成员。在一些实施例中,“动物”是指处于任何发育阶段的人类。在一些实施例中,“动物”指处于任何发育阶段的非人动物。在某些实施例中,非人动物是哺乳动物(例如啮齿类动物、小鼠、大鼠、兔、猴、犬、猫、绵羊、牛、灵长类动物和/或猪)。在一些实施例中,动物包括但不限于哺乳动物、鸟类、爬行动物、两栖动物、鱼类、昆虫和/或蠕虫。在一些实施例中,动物可以是转基因动物、基因工程改造的动物和/或克隆体。
大约或约:如本文所用,当应用于一个或多个所关注的值时,术语“大约”或“约”是指与所述参考值相似的值。在某些实施例中,术语“大约”或“约”是指在所述参考值的任一方向上(大于或小于)落入25%、20%、19%、18%、17%、16%、15%、14%、13%、12%、11%、10%、9%、8%、7%、6%、5%、4%、3%、2%、1%或更小内的一系列值,除非另有说明或从上下文中另外显而易见(此类数字将超过可能值的100%的情况除外)。
递送:如本文所用,术语“递送”涵盖局部递送和全身递送两者。例如,mRNA的递送涵盖其中将mRNA递送至靶组织并且编码的蛋白质在靶组织内表达且保留的情况(也称为“局部分布”或“局部递送”),以及其中将mRNA递送至靶组织并且编码的蛋白质表达且分泌到患者的循环系统(例如血清)内,并且全身分布且被其它组织吸收的情况(也称为“全身分布”或“全身递送”)。
囊封:如本文所用,术语“囊封”或语法等效物是指在纳米颗粒内限制mRNA分子的过程。
表达:如本文所用,核酸序列的“表达”是指将mRNA翻译成肽、多肽或蛋白质,将多个多肽(例如抗体的重链或轻链)组装成完整蛋白质(例如抗体)和/或对多肽或完全组装的蛋白质(例如抗体)进行翻译后修饰。在本申请中,术语“表达”和“产生”以及语法等效物可互换使用。
功能性:如本文所用,“功能性”生物分子为呈显示其特性化特性和/或活性的形式的生物分子。
半衰期:如本文所用,术语“半衰期”为一定数量,例如核酸或蛋白质浓度或活性降至如在时间段开始时测量的其值的一半所需的时间。
改善、增加或减少:如本文所用,术语“改善”、“增加”或“减少”或语法等效物指示相对于基线测量的值,例如在本文所述的治疗开始之前在相同个体中的测量,或在不存在本文所述的治疗的情况下在对照个体(或多个对照个体)中的测量。“对照个体”是罹患与正接受治疗的个体相同形式的疾病的个体,其与正接受治疗的个体的年龄大约相同。
杂质:如本文所用,术语“杂质”是指受限量的液体、气体或固体内的物质,其不同于目标材料或化合物的化学组成。杂质也被称为污染物。
体外:如本文所用,术语“体外”是指在人造环境中,例如在试管或反应容器中,在细胞培养物中等,而不是在多细胞生物体内发生的事件。
体内:如本文使用的,术语“体内”是指在多细胞生物内,例如人类和非人动物内发生的事件。在基于细胞的系统的情况下,所述术语可用于指在活细胞内(与例如体外系统相反)发生的事件。
分离的:如本文所用,术语“分离的”是指如下的物质和/或实体:(1)已与最初产生时与其结合的至少一些组分分离(无论在自然界中和/或在实验环境中),和/或(2)通过人力产生、制备和/或制造。分离的物质和/或实体可与约10%、约20%、约30%、约40%、约50%、约60%、约70%、约80%、约90%、约91%、约92%、约93%、约94%、约95%、约96%、约97%、约98%、约99%或超过约99%的最初与其结合其它组分分离。在一些实施例中,分离的试剂的纯度为约80%、约85%、约90%、约91%、约92%、约93%、约94%、约95%、约96%、约97%、约98%、约99%或超过约99%。如本文所用,如果物质基本上不含其它组分,则其为“纯的”。如本文所用,分离的物质和/或实体的纯度百分比的计算不应包括赋形剂(例如缓冲液、溶剂、水等)。
局部分布或递送:如本文所用,术语“局部分布”、“局部递送”或语法等效物是指组织特异性递送或分布。通常,局部分布或递送需要由mRNA编码的蛋白质或肽在细胞内或伴随有限分泌被翻译和表达,其避免进入患者的循环系统。
信使RNA(mRNA):如本文所用,术语“信使RNA(mRNA)”是指编码至少一种肽、多肽或蛋白质的多核苷酸。如本文所用的mRNA涵盖经修饰的RNA和未修饰的RNA两者。mRNA可含有一个或多个编码和非编码区。mRNA可从天然来源纯化,使用重组表达系统产生且任选地纯化,化学合成等。适当时,例如在化学合成的分子的情况下,mRNA可包含核苷类似物,如具有化学修饰的碱基或糖、主链修饰等的类似物。除非另外指明,否则mRNA序列以5'至3'方向呈现。在一些实施例中,mRNA是或包含天然核苷(例如腺苷、鸟苷、胞苷、尿苷);核苷类似物(例如2-氨基腺苷、2-硫代胸苷、肌苷、吡咯并嘧啶、3-甲基腺苷、5-甲基胞苷、C-5丙炔基-胞苷、C-5丙炔基-尿苷、2-氨基腺苷、C5-溴尿苷、C5-氟尿苷、C5-碘尿苷、C5-丙炔基-尿苷、C5-丙炔基-胞苷、C5-甲基胞苷、2-氨基腺苷、7-脱氮腺苷、7-脱氮鸟苷、8-氧代腺苷、8-氧代鸟苷、O(6)-甲基鸟嘌呤和2-硫代胞苷);化学修饰的碱基;生物学修饰的碱基(例如甲基化碱基);插入碱基;经修饰的糖(例如2'-氟核糖、核糖、2'-脱氧核糖、阿拉伯糖和己糖);和/或经修饰的磷酸基(例如硫代磷酸酯和5'-N-亚磷酰胺键)。
在一些实施例中,mRNA包含一种或多种非标准核苷酸残基。非标准核苷酸残基可包括例如5-甲基-胞苷(“5mC”)、假尿苷(“__U”)和/或2-硫代-尿苷(“2sU”)。关于此类残基和其并入至mRNA中的论述,参见例如美国专利第8,278,036号或WO2011012316。mRNA可以是RNA,其被定义为其中25%的U残基是2-硫代-尿苷且25%的C残基是5-甲基胞苷。使用RNA的教示公开于美国专利公开案US2012/0195936和国际公开案WO 2011/012316中,其均以全文引用方式并入本文。非标准核苷酸残基的存在可使得mRNA比具有相同序列但仅含有标准残基的对照mRNA更稳定和/或免疫原性更小。在另外的实施例中,mRNA可包含选自以下的一种或多种非标准核苷酸残基:异胞嘧啶、假异胞嘧啶、5-溴尿嘧啶、5-丙炔基尿嘧啶、6-氨基嘌呤、2-氨基嘌呤、肌苷、二氨基嘌呤和2-氯-6-氨基嘌呤胞嘧啶,以及这些修饰和其它核碱基修饰的组合。某些实施例可进一步包括对呋喃糖环或核碱基的另外修饰。另外修饰可包括例如糖修饰或取代(例如2'-O-烷基修饰、锁核酸(LNA)中的一个或多个)。在一些实施例中,RNA可与另外的多核苷酸和/或肽多核苷酸(PNA)复合或杂交。在糖修饰是2'-O-烷基修饰的实施例中,此类修饰可包括但不限于2'-脱氧-2'-氟修饰、2'-O-甲基修饰、2'-O-甲氧基乙基修饰和2'-脱氧修饰。在某些实施例中,这些修饰中的任一个可单独或组合地存在于0-100%的核苷酸-例如大于0%、1%、10%、25%、50%、75%、85%、90%、95%或100%的成分核苷酸中。
核酸:如本文所用,术语“核酸”在其最广泛意义上是指并入或可并入多核苷酸链中的任何化合物和/或物质。在一些实施例中,核酸是经由磷酸二酯键并入或可并入多核苷酸链中的化合物和/或物质。在一些实施例中,“核酸”是指个别核酸残基(例如核苷酸和/或核苷)。在一些实施例中,“核酸”是指包含个别核酸残基的多核苷酸链。在一些实施例中,“核酸”涵盖RNA以及单链和/或双链DNA和/或cDNA。此外,术语“核酸”、“DNA”、“RNA”和/或类似术语包括核酸类似物,即具有除磷酸二酯主链以外的类似物。例如,所谓的“肽核酸”,其在所属领域中已知并且在主链中具有肽键而不是磷酸二酯键,被视为在本发明的范围内。术语“编码氨基酸序列的核苷酸序列”包括所有互为简并形式和/或编码相同氨基酸序列的核苷酸序列。编码蛋白质的核苷酸序列和/或RNA可包括内含子。核酸可从天然来源纯化,使用重组表达系统产生且任选地纯化,化学合成等。适当时,例如在化学合成的分子的情况下,核酸可包含核苷类似物,例如具有化学修饰的碱基或糖、主链修饰等的类似物。除非另外指明,否则核酸序列以5'至3'方向呈现。在一些实施例中,核酸是或包含天然核苷(例如腺苷、胸苷、鸟苷、胞苷、尿苷、脱氧腺苷、脱氧胸苷、脱氧鸟苷和脱氧胞苷);核苷类似物(例如2-氨基腺苷、2-硫代胸苷、肌苷、吡咯并嘧啶、3-甲基腺苷、5-甲基胞苷、C-5丙炔基-胞苷、C-5丙炔基-尿苷、2-氨基腺苷、C5-溴尿苷、C5-氟尿苷、C5-碘尿苷、C5-丙炔基-尿苷、C5-丙炔基-胞苷、C5-甲基胞苷、2-氨基腺苷、7-脱氮腺苷、7-脱氮鸟苷、8-氧代腺苷、8-氧代鸟苷、O(6)-甲基鸟嘌呤和2-硫代胞苷);化学修饰的碱基;生物学修饰的碱基(例如甲基化碱基);插入碱基;经修饰的糖(例如2'-氟核糖、核糖、2'-脱氧核糖、阿拉伯糖和己糖);和/或经修饰的磷酸基(例如硫代磷酸酯和5'-N-亚磷酰胺键)。在一些实施例中,本发明特别涉及“未修饰的核酸”,其意指未经化学修饰以便促进或实现递送的核酸(例如,多核苷酸和残基,包括核苷酸和/或核苷)。
患者:如本文所用,术语“患者”或“个体”是指可例如出于实验、诊断、预防、美容和/或治疗目的向其施用所提供的组合物的任何生物体。典型的患者包括动物(例如哺乳动物,如小鼠、大鼠、兔、非人灵长类动物和/或人类)。在一些实施例中,患者是人类。人类包括出生前和出生后形式。
药学上可接受的:如本文所用,术语“药学上可接受的”是指在合理医学判断范围内适用于与人类和动物的组织接触而无过度毒性、刺激、过敏反应或其它问题或并发症,与合理效益/风险比相称的物质。
药学上可接受的盐:药学上可接受的盐是所属领域众所周知的。例如,S.M.Berge等人在《药物科学杂志(J.Pharmaceutical Sciences)》(1977)66:1-19中详细描述了药学上可接受的盐。本发明化合物的药学上可接受的盐包括源自合适的无机和有机酸和碱的那些盐。药学上可接受的无毒性酸加成盐的实例是氨基与无机酸(如盐酸、氢溴酸、磷酸、硫酸和过氯酸)或有机酸(如乙酸、草酸、顺丁烯二酸、酒石酸、柠檬酸、丁二酸或丙二酸)形成的盐,或通过使用所属领域中所用的其它方法(如离子交换)形成的盐。其它药学上可接受的盐包括己二酸盐、藻酸盐、抗坏血酸盐、天冬氨酸盐、苯磺酸盐、苯甲酸盐、硫酸氢盐、硼酸盐、丁酸盐、樟脑酸盐、樟脑磺酸盐、柠檬酸盐、环戊烷丙酸盐、二葡糖酸盐、十二烷基硫酸盐、乙磺酸盐、甲酸盐、富马酸盐、葡庚糖酸盐、甘油磷酸盐、葡糖酸盐、半硫酸盐、庚酸盐、己酸盐、氢碘酸盐、2-羟基-乙磺酸盐、乳糖酸盐、乳酸盐、月桂酸盐、月桂基硫酸盐、苹果酸盐、马来酸盐、丙二酸盐、甲磺酸盐、2-萘磺酸盐、烟酸盐、硝酸盐、油酸盐、草酸盐、棕榈酸盐、双羟萘酸盐、果胶酸盐、过硫酸盐、3-苯基丙酸盐、磷酸盐、苦味酸盐、特戊酸盐、丙酸盐、硬脂酸盐、丁二酸盐、硫酸盐、酒石酸盐、硫氰酸盐、对甲苯磺酸盐、十一烷酸盐、戊酸盐等。源自适当碱的盐包括碱金属盐、碱土金属盐、铵盐和N+(C1-4烷基)4盐。代表性的碱金属盐或碱土金属盐包括钠、锂、钾、钙、镁等。另外的药学上可接受的盐包括(适当时)无毒铵、季铵和使用抗衡离子形成的胺阳离子,例如卤化物、氢氧化物、羧酸盐、硫酸盐、磷酸盐、硝酸盐、磺酸盐和芳基磺酸盐。另外的药学上可接受的盐包括使用适当的亲电子试剂(例如卤代烷)的胺季铵化形成的盐,以形成季铵化的烷基化氨基盐。
个体:如本文所用,术语“个体”是指人类或任何非人动物(例如小鼠、大鼠、兔、犬、猫、牛、猪、绵羊、马或灵长类动物)。人类包括出生前和出生后形式。在许多实施例中,个体是人类。个体可为患者,其是指呈现给医疗服务提供者以便诊断或治疗疾病的人类。术语“个体(subject)”在本文中与“个体(individual)”或“患者”可互换使用。个体可能罹患或易患疾病或病症(例如,囊性纤维化),但可能或可能不显示疾病或病症的症状。
基本上:如本文中所用,术语“基本上”是指展现全部或接近全部范围或程度的所关注的特征或特性的定性条件。生物学领域的普通技术人员应理解,生物学和化学现象很少(如果有的话)达到完全和/或进行到完全或者实现或避免绝对结果。术语“基本上”因此在本文中用于捕获许多生物学现象和化学现象中所固有的潜在完整性缺乏。
全身分布或递送:如本文所用,术语“全身分布”、“全身递送”或语法等效物是指影响整个身体或整个生物体的递送或分布机制或方法。通常,全身分布或递送是经由身体的循环系统,例如血流来完成。与“局部分布或递送”的定义相比。
靶组织:如本文所用,术语“靶组织”是指受待治疗的疾病影响的任何组织。在一些实施例中,靶组织包括展示疾病相关病理学、症状或特征的那些组织。
转移媒剂:在一些实施例中,转移媒剂是脂质体囊泡,或促进核酸转移到靶细胞和组织的其它方式。合适的转移媒剂包括但不限于脂质体、纳米脂质体、含神经酰胺的纳米脂质体、蛋白脂质体、纳米颗粒、磷硅酸钙纳米颗粒、磷酸钙纳米颗粒、二氧化硅纳米颗粒、纳米晶微粒、半导体纳米颗粒、聚(D-精氨酸)、纳米树状体、基于淀粉的递送系统、胶束、乳液、泡囊、质体、病毒、磷酸钙核苷酸、适体、肽和其它矢量标签。还涵盖使用生物纳米胶囊和其它病毒衣壳蛋白装配作为合适的转移媒剂。(《人类基因治疗(Hum.Gene Ther.)2008年9月;19(9):887-95)。
治疗有效量:如本文所用,术语治疗剂的“治疗有效量”意指当施用于患有或易患疾病、病症和/或病况的个体时,足以治疗、诊断、预防和/或延迟疾病、病症和/或病况的症状发作的量。所属领域的普通技术人员应了解,治疗有效量通常经由包含至少一个单位剂量的给药方案施用。
治疗:如本文使用的,术语“治疗(treat/treatment/treating)”是指用于部分或完全减轻、改善、缓解、抑制、预防、延迟特定疾病、病症和/或病况的一种或多种症状或特征的发作、减轻其严重性和/或减少其发生率的任何方法。治疗可施用于未表现出疾病征兆和/或仅表现出疾病早期征兆的个体,用于降低与疾病相关的病理发展的风险。
产率:如本文所用,术语“产率”是指与作为起始物质的总mRNA相比在囊封后回收的mRNA的百分比。在一些实施例中,术语“回收”可与术语“产率”互换使用。
本发明尤其提供使用并有如本文所述的基于固醇的阳离子脂质、辅助脂质和PEG或PEG改性脂质的改进的脂质体在体内递送mRNA的方法和组合物。
脂质纳米颗粒
根据本发明,本文所述的合适的组合物包含核酸和囊封核酸的脂质纳米颗粒,其中所述脂质纳米颗粒包含不同的脂质组分。在一些实施例中,存在不超过三种不同的脂质组分且示例性的不同的脂质组分包括基于固醇的阳离子脂质、辅助脂质(例如非阳离子脂质)和PEG改性脂质。
在一些实施例中,脂质纳米颗粒具有至少约50%、约55%、约60%、约65%、约70%、约75%、约80%、约85%、约90%或约95%的核酸囊封百分比。
在一些实施例中,组合物中囊封核酸的脂质纳米颗粒的百分比为至少约50%、约55%、约60%、约65%、约70%、约75%、约80%、约85%、约90%或约95%。
基于固醇的阳离子脂质
如本文所用,短语“基于固醇的脂质”或“基于固醇的阳离子脂质”是指包含类固醇部分的阳离子脂质。如本文所用,短语“阳离子脂质”是指在所选pH(例如生理pH)下具有净正电荷的多种脂质物质中的任一种。
在实施例中,基于固醇的阳离子脂质具有根据式(A)的结构,
B-L1-S(式A),或其质子化形式。B是碱性官能团(例如,其中质子化形式具有不超过约8.0的pKa)。
L1是任选经取代的连接基团(例如,C1-C20亚烷基或2至20元亚杂烷基)。
S是固醇。
在一些实施例中,B是碱性官能团,其中质子化形式具有不超过约9.0、约8.5、约8.0、约7.5或约7.0的pKa。在实施例中,B是碱性官能团,其中质子化形式具有不超过约8.0或约7.5的pKa。
在一些实施例中,B是碱性官能团,其中质子化形式具有约4.0至约9.0、约4.0至约8.0、约4.5至约8.0、约5.0至约8.0、约5.5至约8.0或约6.0至约8.0的pKa。
在一些实施例中,B是任选经取代的5元或6元含氮杂芳基。
在一些实施例中,B是选自吡咯基、咪唑基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。对于本文所述的任何任选经取代的基团(例如,具有0、1、2、3或4个独立取代基的基团),示例性取代基包括但不限于:卤素(例如-F、-Cl、-Br或-I)、C1-20烷基(例如,C1-6烷基)、C1-20卤烷基(例如,C1-6卤烷基)、-CN、-OH、-O(C1-20烷基)(例如-O(C1-6烷基))、-O(C1-20卤烷基)(例如-O(C1-6卤烷基))、-NH2、-NH(C1-6烷基)和-NH(C1-6烷基)2。
在一些实施例中,B是选自吡咯基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
在一些实施例中,B是选自吡咯基、吡唑基、三唑基、四唑基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
在一些实施例中,L1是连接基团,其为任选经取代的C1-C20亚烷基(例如任选经取代的C1-C10亚烷基或任选经取代的C1-C5亚烷基)。如本文所用,术语“亚烷基”表示饱和二价直链或支链烃基并且由亚甲基、亚乙基、亚异丙基等例示。
在一些实施例中,L1是连接基团,其为任选经取代的2至20元亚杂烷基(例如,任选经取代的2至12元亚杂烷基或任选经取代的2至6元亚杂烷基)。如本文所用,术语“亚杂烷基”是指二价杂烷基。术语“杂烷基”是指具有所陈述数目的原子的稳定直链或支链烃基,其中原子选自碳和一到三个选自由O、N、Si和S组成的群组的杂原子,且其中氮和硫原子可任选地被氧化并且氮杂原子可任选地被季铵化。杂原子O、N和S可置于杂烷基的任何内部位置处。
在一些实施例中,L1是作为任选经取代的2至20元亚杂烷基的连接基团,并且其中L1是非肽的(即非肽连接基团L1是不包含氨基酸残基的连接基团)。
在一些实施例中,L1是连接基团,其包含为酯基、酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。
在一些实施例中,L1是连接基团,其包含为酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。
在一些实施例中,L1是连接基团,其包含为酰胺基、碳酸酯基或脲基的部分。
在一些实施例中,L1是由下式表示的连接基团:
-X1-C(X3)-X2、-(C1-C19亚烷基)-X1-C(X3)-X2、-X1-C(X3)-X2(C1-C19亚烷基)-、-(C1-C19亚烷基)-X1-、-X1-(C1-C19亚烷基)-。
每个X1和X2独立地为共价键、-O-、-S-或-NH-。
X3独立地是=O、=S或=NH。
C1-C19亚烷基(例如,C1-C5亚烷基或C1-C10亚烷基)独立地任选经取代。
在一些实施例中,L1未经取代。
在一些实施例中,L1不包含具有结构-N(R')2的取代基,或其带正电形式,其中每个R'独立地为氢或任选经取代的C1-C20烷基。
在一些实施例中,S是动物固醇,或其氧化或还原形式。在实施例中,S是动物固醇。在实施例中,S是动物固醇的氧化形式。在实施例中,S是动物固醇的还原形式。
在一些实施例中,S是植物固醇,或其氧化或还原形式。在实施例中,S是植物固醇。在实施例中,S是植物固醇的氧化形式。在实施例中,S是植物固醇的还原形式。
在一些实施例中,S是合成固醇(例如,非天然存在的),或其氧化或还原形式。在实施例中,S是合成固醇。在实施例中,S是合成固醇的氧化形式。在实施例中,S是合成固醇的还原形式。
在一些实施例中,S是如本文所述的固醇的氧化形式。
在一些实施例中,固醇的氧化形式是其中母体固醇已经修饰以包括另外的含氧基团的形式。在实施例中,与母体固醇相比,固醇的氧化形式包括一个或多个(例如1、2、3或4个)另外的羟基和/或含羰基基团(例如,酮、醛、羧酸或羧酯部分)。
在一些实施例中,固醇的氧化形式是其中母体固醇已经修饰以包括不饱和碳-碳键(例如碳-碳双键)的形式。在实施例中,与母体固醇相比,固醇的氧化形式包括一个或多个(例如1、2或3个)另外的碳-碳双键。
在一些实施例中,S是如本文所述的固醇的还原形式。
在一些实施例中,固醇的还原形式是其中母体固醇已经修饰以包括较少含氧基团的形式。在实施例中,与母体体醇相比,固醇的还原形式包括减少数目(例如少1、2、3或4个部分)的羟基和/或含羰基基团(例如,酮、醛、羧酸或羧酯部分)。
在一些实施例中,固醇的还原形式是其中母体固醇已经修饰以包括较少不饱和碳-碳键(例如碳-碳双键)的形式。在实施例中,与母体固醇相比,固醇的还原形式包括减少数目(例如少1、2或3个)的碳-碳双键。
在一些实施例中,S是以下固醇:胆固醇、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇或谷固醇,或其任何氧化或还原形式。
在一些实施例中,S是选自胆固醇、胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。
在一些实施例中,S是选自胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。
在一些实施例中,S是以下固醇:胆固醇、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇或谷固醇。
在一些实施例中,S是以下固醇的氧化形式:胆固醇、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇或谷固醇。
在一些实施例中,S是以下固醇的还原形式:胆固醇、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇或谷固醇。
在一些实施例中,S是以下固醇:胆固醇、胆固醇的氧化形式或胆固醇的还原形式。
在一些实施例中,S是以下固醇:烷基石胆酸盐、烷基石胆酸盐的氧化形式或烷基石胆酸盐的还原形式。
在一些实施例中,S是以下固醇:豆固醇、豆固醇的氧化形式或豆固醇的还原形式。
在一些实施例中,S是以下固醇:豆固烷醇、豆固烷醇的氧化形式或豆固烷醇的还原形式。
在一些实施例中,S是以下固醇:菜油固醇、菜油固醇的氧化形式或菜油固醇的还原形式。
在一些实施例中,S是以下固醇:麦角固醇、麦角固醇的氧化形式或麦角固醇的还原形式。
在一些实施例中,S是以下固醇:谷固醇、谷固醇的氧化形式或谷固醇的还原形式。
在一些实施例中,S是以下固醇:胆酸或其烷基酯,或其氧化形式,或其还原形式。
在一些实施例中,S是以下固醇:胆酸或其烷基酯,或其氧化形式,或其还原形式。
在一些实施例中,S是选自以下的固醇:
(胆酸),其中R是任选经取代的C1-C20烷基,且R'是H或任选经取代的C1-C20烷基。
在一些实施例中,S是以下固醇:
在一些实施例中,S是选自以下的固醇:
其中R是任选
经取代的C1-C20烷基,且R'是H或任选经取代的C1-C20烷基。
在一些实施例中,基于固醇的阳离子脂质包含咪唑胆固醇酯(ICE)。
在一些实施例中,基于固醇的阳离子脂质不包含唑胆固醇酯(ICE)。
在一些实施例中,基于固醇的阳离子脂质是基于胆固醇的阳离子脂质。此类基于胆固醇的阳离子脂质可单独使用或与如本文所述的其它脂质组合使用。合适的基于胆固醇的阳离子脂质包括例如DC-Chol(N,N-二甲基-N-乙基甲酰胺基胆固醇)、1,4-双(3-N-油基氨基-丙基)哌嗪(Gao等人《生物化学与生物物理研究通讯(Biochem.Biophys.Res.Comm.)》179,280(1991);Wolf等人《生物技术(BioTechniques)》23,139(1997);美国专利第5,744,335号)。
在实施例中,基于固醇的阳离子脂质是含二烷基氨基、咪唑和胍盐的基于固醇的阳离子脂质。例如,某些实施例涉及包含一种或多种基于固醇的阳离子脂质的组合物,所述阳离子脂质包含咪唑,例如咪唑胆固醇酯或“ICE”脂质3-(1H-咪唑-4-基)丙酸(3S,10R,13R,17R)-10,13-二甲基-17-((R)-6-甲基庚-2-基)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊[a]菲-3-酯,如下文结构(I)所示。在某些实施例中,用于递送编码功能蛋白的RNA(例如mRNA)的脂质纳米颗粒可包含一种或多种基于咪唑的阳离子脂质,例如咪唑胆固醇酯或“ICE”脂质3-(1H-咪唑-4-基)丙酸(3S,10R,13R,17R)-10,13-二甲基-17-((R)-6-甲基庚-2-基)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊[a]菲-3-酯,如下文结构(I)所示。
结构(I)的示例性水解反应提供于方案1中。
方案1
不希望受特定理论束缚,相信基于咪唑的阳离子脂质ICE的融合性与由咪唑基团促进的内体破坏有关,所述咪唑基团具有相对于传统阳离子脂质更低的pKa。内体破坏继而促进渗透溶胀和脂质体膜破坏,然后为其中装载或囊封的多核苷酸内容物转染或细胞内释放至靶细胞中。
基于咪唑的阳离子脂质的特征还在于其相对于其它阳离子脂质降低的毒性。在一些实施例中,包含共混药物组合物的脂质纳米颗粒中的一种或多种包含基于咪唑的阳离子脂质,如ICE,以降低此类共混药物组合物中其它更聚毒性的阳离子脂质的相对浓度。基于咪唑的阳离子脂质(例如,ICE)可用作包含共混制剂的一种或多种脂质纳米颗粒中的唯一阳离子脂质,或者可与传统阳离子脂质(例如,DOPE、DC-Chol)、非阳离子脂质、PEG改性脂质和/或辅助脂质组合。阳离子脂质可包含存在于脂质纳米颗粒中的总脂质的约1%至约90%、约2%至约90%、约5%至约90%、约10%至约90%、约15%至约90%、约20%至约90%、约30%至约90%、约40%至约90%、约50%至约90%、约60%至约90%、约70%至约90%、约80%至约90%、约1%至约80%、约1%至约70%、约1%至约60%、约1%至约50%、约1%至约40%、约1%至约30%、约1%至约20%、约1%至约15%、约1%至约10%、约2%至约70%、约5%至约50%、约10%至约40%,或优选地存在于脂质纳米颗粒中的总脂质的约20%至约70%的摩尔比。
不同的脂质组分的比率
在其中脂质纳米颗粒包含三种且不超过三种不同的脂质组分的实施例中,总脂质含量的比率(即,脂质组分(1):脂质组分(2):脂质组分(3))可表示为x:y:z,其中
(y+z)=100-x。
在一些实施例中,“x”、“y”和“z”中的每一个表示三种不同的脂质组分的摩尔百分比,并且比率是摩尔比。
在一些实施例中,“x”、“y”和“z”中的每一个表示三种不同的脂质组分的重量百分比,并且比率是重量比。
在一些实施例中,由变量“x”表示的脂质组分(1)是基于固醇的阳离子脂质。
在一些实施例中,由变量“y”表示的脂质组分(2)是辅助脂质。
在一些实施例中,由变量“z”表示的脂质组分(3)是PEG脂质。
在一些实施例中,表示脂质组分(1)(例如,基于固醇的阳离子脂质)的摩尔百分比的变量“x”为至少约10%、约20%、约30%、约40%、约50%、约55%、约60%、约65%、约70%、约75%、约80%、约85%、约90%或约95%。
在一些实施例中,表示脂质组分(1)(例如,基于固醇的阳离子脂质)的摩尔百分比的变量“x”为不超过约95%、约90%、约85%、约80%、约75%、约70%、约65%、约60%、约55%、约50%、约40%、约30%、约20%或约10%。在一些实施例中,变量“x”为不超过约65%、约60%、约55%、约50%或约40%。
在一些实施例中,表示脂质组分(1)(例如,基于固醇的阳离子脂质)的摩尔百分比的变量“x”为:至少约50%但小于约95%;至少约50%但小于约90%;至少约50%但小于约85%;至少约50%但小于约80%;至少约50%但小于约75%;至少约50%但小于约70%;至少约50%但小于约65%;或至少约50%但小于约60%。
在一些实施例中,表示脂质组分(1)(例如,基于固醇的阳离子脂质)的重量百分比的变量“x”为至少约10%、约20%、约30%、约40%、约50%、约55%、约60%、约65%、约70%、约75%、约80%、约85%、约90%或约95%。
在一些实施例中,表示脂质组分(1)(例如,基于固醇的阳离子脂质)的重量百分比的变量“x”为不超过约95%、约90%、约85%、约80%、约75%、约70%、约65%、约60%、约55%、约50%、约40%、约30%、约20%或约10%。
在一些实施例中,表示脂质组分(1)(例如,基于固醇的阳离子脂质)的重量百分比的变量“x”为:至少约50%但小于约95%;至少约50%但小于约90%;至少约50%但小于约85%;至少约50%但小于约80%;至少约50%但小于约75%;至少约50%但小于约70%;至少约50%但小于约65%;或至少约50%但小于约60%。
在一些实施例中,表示脂质组分(3)(例如,PEG改性脂质)的摩尔百分比的变量“z”为不超过约1%、2%、3%、4%、5%、6%、7%、8%、9%、10%、15%、20%或25%。在一些实施例中,表示脂质组分(3)(例如,PEG改性脂质)的摩尔百分比的变量“z”为约1%、2%、3%、4%、5%、6%、7%、8%、9%、10%。在实施例中,表示脂质组分(3)(例如,PEG改性脂质)的摩尔百分比的变量“z”为约1%至约10%、约2%至约10%、约3%至约10%、约4%至约10%、约5%至约10%、约6%至约10%、约7%至约10%、约8%至约10%、约9%至约10%、约1%至约9%、约1%至约8%、约1%至约7.5%、约1%至约7%、约1%至约6%、约1%至约5%、约1%至约4%、约1%至约3%、约1%至约2%、约2.5%至约10%、约2.5%至约7.5%、约2.5%至约5%、约5%至约7.5%或约5%至约10%。
在一些实施例中,表示脂质组分(3)(例如,PEG改性脂质)的重量百分比的变量“z”为不超过约1%、2%、3%、4%、5%、6%、7%、8%、9%、10%、15%、20%或25%。在一些实施例中,表示脂质组分(3)(例如,PEG脂质)的重量百分比的变量“z”为约1%、2%、3%、4%、5%、6%、7%、8%、9%、10%。在一些实施例中,表示脂质组分(3)(例如,PEG改性脂质)的重量百分比的变量“z”为约1%至约10%、约2%至约10%、约3%至约10%、约4%至约10%、约5%至约10%、约6%至约10%、约7%至约10%、约8%至约10%、约9%至约10%、约1%至约9%、约1%至约8%、约1%至约7.5%、约1%至约7%、约1%至约6%、约1%至约5%、约1%至约4%、约1%至约3%、约1%至约2%、约2.5%至约10%、约2.5%至约7.5%、约2.5%至约5%、约5%至约7.5%或约5%至约10%。
对于具有三种且仅三种不同的脂质组分的组合物,变量“x”、“y”和“z”可呈任何组合,只要三个变量的总和为总脂质含量的100%。
其它脂质组分
本发明也涵盖单独或优选地与包含转移媒剂(例如脂质纳米颗粒)的其它脂质制剂组合在一起使用聚乙二醇(PEG)改性磷脂和衍生的脂质,例如衍生的神经酰胺(PEG-CER),包括N-辛酰基-鞘氨醇-1-[丁二酰基(甲氧基聚乙二醇)-2000](C8PEG-2000神经酰胺)。考虑的PEG改性脂质包括但不限于长度高达5kDa的聚乙二醇链,其与具有C6-C20长度的烷基链的脂质共价连接。此类组分的添加可防止复杂的聚集,并且还可提供用于增加循环寿命和增加脂质-核酸组合物向靶组织的递送的方法(Klibanov等人(1990)《FEBS快报(FEBS Letters)》,268(1):235-237),或其可选择为在体内快速更换制剂(参见美国专利第5,885,613号)。特别有用的可交换脂质是具有较短酰基链(例如,C14或C18)的PEG-神经酰胺。
本发明的PEG改性磷脂和衍生的脂质可包含存在于脂质体转移媒剂中的总脂质的约0%至约20%、约0.5%至约20%、约1%至约20%、约2%至约20%、约3%至约20%、约4%至约20%、约5%至约20%、约10%至约20%、约15%至约20%、约0%至约15%、约0%至约10%、约0%至约5%、约0%至约4%、约0%至约3%、约0%至约220%、约0%至约1%、约5%至约15%、约4%至约10%或约2%的摩尔比。
本发明还涵盖非阳离子脂质的用途。如本文所用,短语“非阳离子脂质”是指任何中性、两性离子或阴离子脂质。如本文所用,短语“阴离子脂质”是指在所选pH(例如生理pH)下具有净负电荷的多种脂质物质中的任一种。非阳离子脂质包括但不限于二硬脂酰基磷脂酰胆碱(DSPC)、二油酰基磷脂酰胆碱(DOPC)、二棕榈酰基磷脂酰胆碱(DPPC)、二油酰基磷脂酰甘油(DOPG)、二棕榈酰磷脂酰甘油(DPPG)、二油酰基磷脂酰乙醇胺(DOPE)、棕榈酰油酰基磷脂酰胆碱(POPC)、棕榈酰油酰基-磷脂酰乙醇胺(POPE)、二油酰基-磷脂酰乙醇胺4-(N-马来酰亚胺基甲基)-环己烷-1-甲酸酯(DOPE-mal)、二棕榈酰基磷脂酰乙醇胺(DPPE)、二肉豆蔻酰基磷脂酰乙醇胺(DMPE)、二硬脂酰基-磷脂酰基-乙醇胺(DSPE)、16-O-单甲基PE,16-O-二甲基PE,18-1-反式PE,1-硬脂酰基-2-油酰基-磷脂酰乙醇胺(SOPE)、胆固醇或其混合物。此类非阳离子脂质可单独使用,但优选与其它赋形剂,例如阳离子脂质组合使用。
在一些实施例中,并非基于固醇的阳离子脂质的阳离子脂质可用于本发明的脂质体组合物中。阳离子脂质包括但不限于N-[1-(2,3-二油基氧基)丙基]-N,N,N-三甲基氯化铵(DOTMA)、5-羧基精胺基甘氨酸双十八烷基酰胺(DOGS)、2,3-二油基氧基-N-[2(精胺-甲酰胺基)乙基]-N,N-二甲基-1-丙铵盐(DOSPA)、1,2-二油酰基-3-二甲基铵-丙烷(DODAP)、1,2-二油酰基-3-三甲基铵-丙烷(DOTAP)、1,2-二硬脂酰氧基-N,N-二甲基-3-氨基丙烷(DSDMA)、1,2-二油基氧基-N,N-二甲基-3-氨基丙烷(DODMA)、1,2-二亚油醇基氧基-N,N-二甲基-3-氨基丙烷(DLinDMA)、1,2-二亚麻醇基氧基-N,N-二甲基-3-氨基丙烷(DLenDMA)、N-二油基-N,N-二甲基氯化铵(DODAC)、N,N-二硬脂基-N,N-二甲基溴化铵(DDAB)、N-(1,2-二肉豆蔻基氧基丙-3-基)-N,N-二甲基-N-羟乙基溴化铵(DMRIE)、3-二甲氨基-2-(胆甾-5-烯-3-β-氧基丁-4-氧基)-1-(顺式,顺式-9,12-十八碳二烯氧基)丙烷(CLinDMA)、2-[5'-(胆甾-5-烯-3-β-氧基)-3'-氧杂戊氧基)-3-二甲基1-1-(顺式,顺式-9',1-2'-十八碳二烯氧基)丙烷(CpLinDMA)、N,N-二甲基-3,4-二油基氧基苯甲胺(DMOBA)、1,2-N,N'-二油基氨甲酰基-3-二甲基氨基丙烷(DOcarbDAP)、2,3-二亚油酰氧基-N,N-二甲基丙胺(DLinDAP)、1,2-N,N'-二亚油醇基氨甲酰基-3-二甲基氨基丙烷(DLincarbDAP)、1,2-二亚油酰基氨甲酰基-3-二甲基氨基丙烷或(DLinCDAP)、2,2-亚油醇基-4-二甲氨基甲基-[1,3]-二氧杂环戊烷(DLin-K-DMA)、2,2-亚油醇基-4-二甲氨基乙基-[1,3]-二氧杂环戊烷(DLin-K-XTC2-DMA),或其混合物。
另外的阳离子脂质包括2-((2,3-双((9Z,12Z)-十八-9,12-二烯-1-基氧基)丙基)二硫烷基)-N,N-二甲基乙胺(进一步描述于2012年6月8日提交的美国临时申请第PCT/US2012/041663号中,其全部教示内容以全文引用的方式并入本文中)、可裂解脂质,例如一种或多种包含可裂解二硫键(S-S)官能团的阳离子脂质,如美国国际申请第PCT/US2012/041663号中所进一步描述。另外,若干试剂可商购获得以增强转染功效。合适的实例包括LIPOFECTIN(DOTMA:DOPE)(Invitrogen,Carlsbad,Calif.)、LIPOFECTAMINE(DOSPA:DOPE)(Invitrogen)、LIPOFECTAMINE2000(Invitrogen)、FUGENE、TRANSFECTAM(DOGS)和EFFECTENE。
囊封mRNA的脂质体的形成
用于本发明的脂质体转移媒剂可通过所属领域目前已知的各种技术制备。多层囊泡(MLV)可根据常规技术制备,例如通过将所选择的脂质沉积在合适的容器或器皿的内壁上,其通过将脂质溶解在适当溶剂中,且接着蒸发溶剂以在器皿的内部上留下薄膜或通过喷雾干燥。然后可用涡旋运动将水相添加至器皿中,这导致MLV的形成。然后可通过多层囊泡的均质化、超声处理或挤出来形成单层囊泡(ULV)。另外,可通过清洁剂去除技术形成单层囊泡。
在本发明的某些实施例中,本发明组合物包含转移媒剂,其中治疗性RNA(例如编码CFTR的mRNA)在转移媒剂(例如脂质体)的两个表面上结合,且囊封在相同转移媒剂内。例如,在本发明组合物的制备过程中,阳离子脂质体转移媒剂可通过与此类治疗性mRNA的静电相互作用而与核酸(例如mRNA)结合。
在一些实施例中,本发明的组合物和方法包含囊封在脂质体中的mRNA。在一些实施例中,一种或多种mRNA物种可囊封在相同的脂质体中。在一些实施例中,一种或多种mRNA物种可囊封在不同的脂质体中。在一些实施例中,mRNA囊封在一种或多种脂质体中,所述脂质体在其脂质组成、脂质组分的摩尔比、尺寸、电荷(ζ电位)、靶向配体和/或其组合方面不同。在一些实施例中,一种或多种脂质体可具有基于固醇的阳离子脂质、中性脂质、PEG改性脂质和/或其组合的不同组成。在一些实施例中,一种或多种脂质体可具有用于产生脂质体的基于固醇的阳离子脂质、中性脂质和PEG改性脂质的不同摩尔比。
将所需mRNA并入脂质体内的过程通常称为“加载”。示例性方法在Lasic等人,《欧洲生物化学学会联合会快报(FEBS Lett.)》,312:255-258,1992中描述,所述参考文献以引用的方式并入本文。并有脂质体的核酸可完全或部分地位于脂质体的内部空间中、脂质体的双层膜内、或与脂质体膜的外表面结合。将核酸并入脂质体中在本文中也称为“囊封”,其中核酸完全包含在脂质体的内部空间内。将mRNA并入转移媒剂(例如脂质体)中的目的经常是保护核酸免受环境侵害,所述环境可能含有降解核酸的酶或化学物质和/或促使核酸快速排泄的系统或受体。因此,在一些实施例中,合适的递送媒剂能够增强其中包含的mRNA的稳定性和/或促进mRNA递送至靶细胞或组织。
任何所需脂质可以任何适合于囊封mRNA的比率混合。在一些实施例中,合适的脂质溶液含有所需脂质的混合物,所需脂质包括基于固醇的阳离子脂质、非阳离子脂质和/或PEG改性脂质。在一些实施例中,合适的脂质溶液含有所需脂质的混合物,所需脂质包括一种或多种基于固醇的阳离子脂质、一种或多种辅助脂质(例如非阳离子脂质)和一种或多种PEG改性脂质。
在实例部分中描述了基于固醇的阳离子脂质、非阳离子脂质和PEG改性脂质的示例性组合。例如,合适的脂质溶液可含有ICE、DOPE和DMG-PEG2K。包含脂质混合物的基于固醇的阳离子脂质、非阳离子脂质和/或PEG改性脂质,以及这些脂质彼此的相对摩尔比的选择是基于所选脂质的特征和待囊封的mRNA的性质和特征。另外的考虑因素包括例如烷基链的饱和度,以及所选脂质的尺寸、电荷、pH、pKa、融合性和毒性。因此,可相应地调节摩尔比。
在一些实施例中,用于将mRNA囊封在脂质纳米颗粒中的方法包含混合mRNA溶液和脂质溶液,其中在混合之前将mRNA溶液和/或脂质溶液加热至高于环境温度的预定温度,以形成囊封mRNA的脂质纳米颗粒(参见2015年7月2日提交的名称为“信使RNA的囊封(Encapsulation of messenger RNA)”的美国专利申请第14/790,562号和2014年7月2日提交的其临时美国专利申请第62/020,163号,所述专利申请的公开内容全文并入本文)。
在一些实施例中,用于将mRNA囊封在脂质纳米颗粒中的方法包含将预形成的脂质纳米颗粒与mRNA组合(参见2016年11月10日提交的美国临时申请第62/420,413号和2017年11月1日提交的美国临时申请第62/580,155号,所述临时申请的公开内容以引用的方式并入本文)。在一些实施例中,将预形成的脂质纳米颗粒与mRNA组合产生脂质纳米颗粒,其显示出改善的细胞内mRNA递送功效。在一些实施例中,将预形成的脂质纳米颗粒与mRNA组合导致脂质纳米颗粒中囊封的mRNA的极高囊封效率(即,在90-95%的范围内)。在一些实施例中,利用泵系统实现将预形成的脂质纳米颗粒与mRNA组合,所述泵系统在整个过程中维持脂质/mRNA(N/P)比恒定,并且还提供了便捷的按比例扩大。
根据本发明的合适的脂质体可以各种尺寸制备。在一些实施例中,提供的脂质体可制备得小于先前已知的囊封mRNA的脂质体。在一些实施例中,脂质体的尺寸减小与更有效的mRNA递送相关。适当脂质体尺寸的选择可考虑到靶细胞或组织的位点,以及在一定程度上制备脂质体所用于的应用。
在一些实施例中,选择适当尺寸的脂质体以促进由mRNA编码的蛋白质或肽的全身分布。在一些实施例中,可能需要限制mRNA向某些细胞或组织的递送。例如,为了靶向肝细胞,脂质体的尺寸可设定为使得其尺寸小于肝脏中内衬肝血窦的内皮层的开窗;在这种情况下,脂质体可容易地穿透这种内皮开窗以到达靶肝细胞。
或者或另外,脂质体的尺寸可设定为使得脂质体的尺寸具有足够的直径,以限制或明确地避免分布到某些细胞或组织内。例如,脂质体的尺寸可设定为使得其尺寸大于内衬肝血窦的内皮层的开窗,从而限制脂质体向肝细胞的分布。
在一些实施例中,脂质体的尺寸由脂质体颗粒的最大直径的长度确定。在一些实施例中,合适的脂质体的尺寸不超过约250nm(例如不超过约225nm、200nm、175nm、150nm、125nm、100nm、75nm或50nm)。在一些实施例中,合适的脂质体的尺寸在约10-250nm范围内(例如在约10-225nm、10-200nm、10-175nm、10-150nm、10-125nm、10-100nm、10-75nm或10-50nm范围内)。在一些实施例中,合适的脂质体的尺寸在约100-250nm范围内(例如在约100-225nm、100-200nm、100-175nm、100-150nm范围内)。在一些实施例中,合适的脂质体的尺寸在约10-100nm范围内(例如在约10-90nm、10-80nm、10-70nm、10-60nm或10-50nm范围内)。在特定实施例中,合适的脂质体具有小于约100nm的尺寸。在一些实施例中,组合物中的大多数纳米颗粒,即大于约50%、55%、60%、65%、70%、75%、80%、85%、90%、95%、96%、97%、98%或99%的纯化的纳米颗粒的尺寸小于约100nm(例如小于约95nm、约90nm、约85nm、约80nm、约75nm、约70nm、约65nm、约60nm、约55nm、约50nm、约45nm或约40nm)。在一些实施例中,基本上所有的纯化纳米颗粒的尺寸小于100nm(例如小于约95nm、约90nm、约85nm、约80nm、约75nm、约70nm、约65nm、约60nm、约55nm、约50nm、约45nm或约40nm)。
所属领域已知的各种替代方法可用于测定脂质体群体的大小。一种这样的尺寸测定方法在美国专利第4,737,323号中描述,所述专利以引用的方式并入本文。通过浴或探针超声处理对脂质体悬浮液进行超声处理产生渐进的尺寸减小,降至直径小于约0.05微米的小ULV。均质化是依赖于剪切能将大脂质体断裂成较小脂质体的另一种方法。在典型的均质化程序中,MLV通过标准乳液均质器再循环,直到观察到选择的脂质体尺寸,通常在约0.1至0.5微米之间。脂质体的尺寸可通过准电光散射(QELS)进行测定,如以引用的方式并入本文的Bloomfield,《生物物理与生物工程年鉴(Ann.Rev.Biophys.Bioeng.)》,10:421-150(1981)中所述。平均脂质体直径可通过所形成的脂质体的超声处理来减小。间歇性超声处理循环可与QELS评价交替,以指导有效的脂质体合成。
纯化
通常,在配制之后,含有mRNA的脂质纳米颗粒经纯化和/或浓缩。可以使用各种纯化方法。在一些实施例中,使用切向流过滤来纯化脂质纳米颗粒。也称为交叉流过滤的切向流过滤(TFF)是一种类型的过滤,其中待过滤材料沿切线方向传递跨越过滤器,而非通过过滤器。在TFF中,非所需渗透物穿过过滤器,而所需渗余物沿着过滤器传递且在下游收集。重要的是,应注意,所需材料通常包含在TFF中的渗余物中,其与传统的死端过滤中通常遇到的情况相反。
取决于待过滤的材料,TFF通常用于微过滤或超过滤。微过滤通常被定义为过滤器具有0.05μm与1.0μm之间的孔尺寸的情况,而超过滤通常包括孔尺寸小于0.05μm的过滤器。孔尺寸还确定标称分子量限值(NMWL),其也称为特定过滤器的分子量截止(MWCO),其中微过滤膜通常具有大于1,000千道尔顿(kDa)的NMWL且超过滤过滤器具有1kDa与1,000kDa之间的NMWL。
切向流过滤的主要优点是在传统的“死端”过滤期间可能聚集在过滤器中且阻塞过滤器的不可渗透颗粒(有时称为“滤饼”)替代地沿着过滤器的表面携带。此优点允许在需要连续操作的工业过程中广泛使用切向流过滤,这是因为停机时间显著减小,因为过滤器通常不需要被移除和清洁。
切向流过滤可用于包括浓缩和渗滤等的若干目的。浓缩是借以从溶液中去除溶剂,同时保留溶质分子的过程。为了有效地浓缩样品,使用具有基本上低于待保留的溶质分子的分子量的NMWL或MWCO的膜。一般来说,技术人员可选择具有低于靶分子的分子量三至六倍的NMWL或MWCO的过滤器。
渗滤是分馏过程,其中小的非所需颗粒穿过过滤器,同时将较大的所需纳米颗粒维持在渗余物中,而不改变那些纳米颗粒的在溶液中的浓度。渗滤通常用于从溶液中去除盐或反应缓冲液。渗滤可以是连续的或不连续的。在连续渗滤中,以与产生滤液相同的速率将渗滤溶液添加到样品进料中。在不连续渗滤中,首先稀释溶液并且接着浓缩回起始浓度。可以重复不连续渗滤,直到达到纳米颗粒的所需浓度。
纯化和/或浓缩的脂质纳米颗粒可以在所需缓冲液(例如PBS)中配制。
因此,本发明提供包含本文所述的纯化纳米颗粒的组合物。在一些实施例中,组合物中的大多数纯化纳米颗粒,即超过约50%、55%、60%、65%、70%、75%、80%、85%、90%、95%、96%、97%、98%或99%的纯化纳米颗粒的尺寸小于约100nm(例如小于约95nm、约90nm、约85nm、约80nm、约75nm、约70nm、约65nm、约60nm、约55nm、约50nm、约45nm或约40nm)。在一些实施例中,基本上所有的纯化纳米颗粒的尺寸小于100nm(例如小于约95nm、约90nm、约85nm、约80nm、约75nm、约70nm、约65nm、约60nm、约55nm、约50nm、约45nm或约40nm)。
在一些实施例中,由本发明提供的组合物中超过约70%、75%、80%、85%、90%、95%、96%、97%、98%、99%的纯化纳米颗粒的尺寸在约40-90nm(例如约40-85nm、约40-80nm、约40-75nm、约40-70nm、约40-65nm或约40-60nm)范围内。在一些实施例中,基本上所有的纯化纳米颗粒的尺寸在约40-90nm(例如约40-85nm、约40-80nm、约40-75nm、约40-70nm、约40-65nm或约40-60nm)范围内。
在一些实施例中,由本发明提供的组合物中的纳米颗粒的分散性或分子尺寸非均质性的量度(PDI)小于约0.16(例如小于约0.15、0.14、0.13、0.12、0.11、0.10、0.09或0.08)。
在一些实施例中,由本发明提供的组合物中超过约75%、80%、85%、90%、95%、96%、97%、98%或99%的纯化脂质纳米颗粒囊封每个单独的颗粒内的mRNA。在一些实施例中,组合物中的基本上所有的纯化脂质纳米颗粒在每个单独的颗粒内囊封mRNA。
在一些实施例中,根据本发明的组合物含有至少约1mg、5mg、10mg、100mg、500mg或1000mg的囊封mRNA。在一些实施例中,根据本发明的过程导致回收超过约60%、65%、70%、75%、80%、85%、90%、95%、96%、97%、98%或99%的mRNA。
核酸和mRNA
本发明可用于递送任何类型的核酸。核酸可选自包含(但不限于)DNA、信使RNA(mRNA)、小核RNA(snRNA)、向导RNA(gRNA)、CRISPR RNA(crRNA)、长非编码RNA(lncRNA)、微小RNA(miRNA)、小干扰RNA(siRNA)和短发夹RNA(shRNA)的群组。
本发明可用于递送任何mRNA。mRNA通常被认为是将信息从DNA携带至核糖体的RNA类型。通常,在真核生物体中,mRNA加工包括在N端(5')末端上添加“帽”,并且在C端(3')末端上添加“尾”。典型的帽是7-甲基鸟苷帽,其为通过5'-5'-三磷酸键连接至第一转录核苷酸的鸟苷。帽的存在对于提供针对大部分真核细胞中发现的核酸酶的抗性是重要的。尾通常是聚腺苷酸化事件,其中聚腺苷酸部分添加至mRNA分子的3'端。此“尾”的存在用以保护mRNA免于核酸外切酶降解。信使RNA通常由核糖体翻译成构成蛋白质或肽的一系列氨基酸。
将任何能够翻译成一种或多种肽(例如抗原)或肽片段的mRNA考虑为在本发明的范围内。在一些实施例中,mRNA编码一种或多种天然存在的肽。在一些实施例中,mRNA编码一种或多种修饰的或非天然的肽。
在一些实施例中,mRNA编码细胞内蛋白或肽。在一些实施例中,mRNA编码胞质蛋白或肽。在一些实施例中,mRNA编码与肌动蛋白细胞骨架相关的蛋白质或肽。在一些实施例中,mRNA编码与质膜相关的蛋白质或肽。在一些具体实施例中,mRNA编码跨膜蛋白或肽。在一些具体实施例中,mRNA编码离子通道蛋白或肽。在一些实施例中,mRNA编码核周蛋白或肽。在一些实施例中,mRNA编码核蛋白或肽。在一些具体实施例中,mRNA编码转录因子。在一些实施例中,mRNA编码伴随蛋白或肽。在一些实施例中,mRNA编码胞内酶(例如编码与脲循环或溶酶体贮积代谢障碍相关的酶的mRNA)。在一些实施例中,mRNA编码涉及细胞代谢、DNA修复、转录和/或翻译的蛋白质或肽。在一些实施例中,mRNA编码细胞外蛋白或肽。在一些实施例中,mRNA编码与细胞外基质相关的蛋白质或肽。在一些实施例中,mRNA编码分泌蛋白或肽。在具体实施例中,本发明的组合物和方法中使用的mRNA可用于表达功能蛋白或酶,所述功能蛋白或酶由一种或多种靶细胞排泄或分泌到周围细胞外液中(例如编码激素和/或神经传递质的mRNA)。
在一些实施例中,本发明的组合物和方法实现了编码分泌蛋白的mRNA的递送。在一些实施例中,本发明的组合物和方法实现了编码表1中列出的一种或多种分泌蛋白的mRNA的递送;因此,本发明的组合物可包含编码表1中列出的蛋白质(或其同源物)的mRNA连同本文所述的其它组分,并且本发明的方法可包含制备和/或施用包含编码表1中列出的蛋白质(或其同源物)的mRNA连同本文所述的其它组分的组合物。
表1.分泌蛋白
在一些实施例中,本发明的组合物和方法实现了一种或多种mRNA的递送,所述mRNA编码表2中列出的一种或多种另外的示例性蛋白;因此,本发明的组合物可包含编码表2中列出的蛋白质(或其同源物)的mRNA连同本文所述的其它组分,并且本发明的方法可包含制备和/或施用包含编码选自表2中列出的蛋白质的蛋白质(或其同源物)的mRNA连同本文所述的其它组分的组合物。
表2.其它示例性蛋白质
表1和表2中阐述的Uniprot ID是指所列蛋白质的人类型式,并且各自的序列可从Uniprot数据库获得。所列蛋白质的序列通常也可用于各种动物,包括各种哺乳动物和兽医或工业感兴趣的动物。因此,在一些实施例中,本发明的组合物和方法实现了一种或多种mRNA的递送,所述mRNA编码一种或多种蛋白质,所述蛋白质选自表1或表2中列出的分泌蛋白的哺乳动物同源物或来自兽医或工业感兴趣的动物的同源物;因此,本发明的组合物可包含编码蛋白质的mRNA连同本文所述的其它组分,所述蛋白质选自表1或表2中列出的蛋白质的哺乳动物同源物或来自兽医或工业感兴趣的动物的同源物,且本发明的方法可包含制备和/或施用包含编码蛋白质的mRNA连同本文所述的其它组分的组合物,所述蛋白质选自表1或表2中列出的蛋白质的哺乳动物同源物或来自兽医或工业感兴趣的动物的同源物。在一些实施例中,哺乳动物同源物选自小鼠、大鼠、仓鼠、沙鼠、马、猪、牛、美洲驼、羊驼、貂、狗、猫、雪貂、绵羊、山羊或骆驼同源物。在一些实施例中,兽医或工业感兴趣的动物选自上文列出的哺乳动物和/或鸡、鸭、火鸡、鲑鱼、鲶鱼或罗非鱼。
在实施例中,本发明的组合物和方法实现了编码选自表3的溶酶体蛋白的mRNA的递送。在一些实施例中,本发明的组合物和方法实现了编码表3中列出的一种或多种溶酶体和/或相关蛋白质的mRNA的递送;因此,本发明的组合物可包含编码表3中列出的蛋白质(或其同源物)的mRNA连同本文所述的其它组分,并且本发明的方法可包含制备和/或施用包含编码表3中列出的蛋白质(或其同源物)的mRNA连同本文所述的其它组分的组合物。
表3.溶酶体和相关蛋白
关于溶酶体蛋白的信息可获自Lubke等人,“溶酶体蛋白质组学(Proteomics ofthe Lysosome)”,生物化学与生物物理学报(Biochim Biophys Acta.)(2009)1793:625-635。在一些实施例中,表3中列出的并且在本发明的组合物和方法中由mRNA编码的蛋白是人蛋白。所列蛋白的序列也可用于各种动物,包括如上所述的各种哺乳动物和兽医或工业感兴趣的动物。
在一些实施例中,本发明的组合物和方法实现了编码如表4中所列的治疗蛋白(例如胞质、跨膜或分泌)的mRNA的递送。在一些实施例中,本发明的组合物和方法实现了编码适用于治疗表4中列出的疾病或病症(即适应症)的治疗蛋白的mRNA的递送;因此,本发明的组合物可包含编码表4中列出或未列出的治疗蛋白(或其同源物,如下文所论述)的mRNA连同本文所述的其它用于治疗表4中列出的疾病或病症(即适应症)的组分,并且本发明的方法可包含制备和/或施用包含编码此类蛋白(或其同源物,如下文所论述)的mRNA连同本文所述的其它用于治疗表4中列出的疾病或病症的组分的组合物。
表4.示例性适应症和相关蛋白
在一些实施例中,本发明用于预防、治疗和/或治愈受疾病或病症影响的个体,所述疾病或病症在表1、2、3或4中列出或与表中列出的蛋白质相关。在一些实施例中,mRNA编码囊性纤维化跨膜传导调节因子(CFTR)、精氨基琥珀酸合成酶(ASS1)、因子IX、生存运动神经元1(SMN1)或苯丙氨酸羟化酶(PAH)中的一种或多种。
在一些实施例中,编码表1、2、3或4中列出的蛋白质中的任一种的mRNA是密码子优化的。在一个实施例中,编码CFTR的mRNA是密码子优化的。
表5.人CFTR
示例性密码子优化的人囊性纤维化跨膜传导调节因子(CFTR)mRNA
构筑体设计:
X-编码序列-Y
5'和3'UTR序列:
X(5'UTR序列)=
GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAGACACGGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGCGGAUUCCCCGUGCCAAGAGUGACUCACCGUCCUUGACACG(SEQID NO:5)
Y(3'UTR序列)=
CGGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCA UCAAGCU(SEQ ID NO:6)
或
GGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAUCAAAGCU(SEQ ID NO:7)
在一个实施例中,密码子优化的人CFTR mRNA序列包括SEQ ID NO:1。
在一个实施例中,全长密码子优化的人CFTR mRNA序列为:
GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAGACACCGGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGCGGAUUCCCCGUGCCAAGAGUGACUCACCGUCCUUGACACGAUGCAACGCUCUCCUCUUGAAAAGGCCUCGGUGGUGUCCAAGCUCUUCUUCUCGUGGACUAGACCCAUCCUGAGAAAGGGGUACAGACAGCGCUUGGAGCUGUCCGAUAUCUAUCAAAUCCCUUCCGUGGACUCCGCGGACAACCUGUCCGAGAAGCUCGAGAGAGAAUGGGACAGAGAACUCGCCUCAAAGAAGAACCCGAAGCUGAUUAAUGCGCUUAGGCGGUGCUUUUUCUGGCGGUUCAUGUUCUACGGCAUCUUCCUCUACCUGGGAGAGGUCACCAAGGCCGUGCAGCCCCUGUUGCUGGGACGGAUUAUUGCCUCCUACGACCCCGACAACAAGGAAGAAAGAAGCAUCGCUAUCUACUUGGGCAUCGGUCUGUGCCUGCUUUUCAUCGUCCGGACCCUCUUGUUGCAUCCUGCUAUUUUCGGCCUGCAUCACAUUGGCAUGCAGAUGAGAAUUGCCAUGUUUUCCCUGAUCUACAAGAAAACUCUGAAGCUCUCGAGCCGCGUGCUUGACAAGAUUUCCAUCGGCCAGCUCGUGUCCCUGCUCUCCAACAAUCUGAACAAGUUCGACGAGGGCCUCGCCCUGGCCCACUUCGUGUGGAUCGCCCCUCUGCAAGUGGCGCUUCUGAUGGGCCUGAUCUGGGAGCUGCUGCAAGCCUCGGCAUUCUGUGGGCUUGGAUUCCUGAUCGUGCUGGCACUGUUCCAGGCCGGACUGGGGCGGAUGAUGAUGAAGUACAGGGACCAGAGAGCCGGAAAGAUUUCCGAACGGCUGGUGAUCACUUCGGAAAUGAUCGAAAACAUCCAGUCAGUGAAGGCCUACUGCUGGGAAGAGGCCAUGGAAAAGAUGAUUGAAAACCUCCGGCAAACCGAGCUGAAGCUGACCCGCAAGGCCGCUUACGUGCGCUAUUUCAACUCGUCCGCUUUCUUCUUCUCCGGGUUCUUCGUGGUGUUUCUCUCCGUGCUCCCCUACGCCCUGAUUAAGGGAAUCAUCCUCAGGAAGAUCUUCACCACCAUUUCCUUCUGUAUCGUGCUCCGCAUGGCCGUGACCCGGCAGUUCCCAUGGGCCGUGCAGACUUGGUACGACUCCCUGGGAGCCAUUAACAAGAUCCAGGACUUCCUUCAAAAGCAGGAGUACAAGACCCUCGAGUACAACCUGACUACUACCGAGGUCGUGAUGGAAAACGUCACCGCCUUUUGGGAGGAGGGAUUUGGCGAACUGUUCGAGAAGGCCAAGCAGAACAACAACAACCGCAAGACCUCGAACGGUGACGACUCCCUCUUCUUUUCAAACUUCAGCCUGCUCGGGACGCCCGUGCUGAAGGACAUUAACUUCAAGAUCGAAAGAGGACAGCUCCUGGCGGUGGCCGGAUCGACCGGAGCCGGAAAGACUUCCCUGCUGAUGGUGAUCAUGGGAGAGCUUGAACCUAGCGAGGGAAAGAUCAAGCACUCCGGCCGCAUCAGCUUCUGUAGCCAGUUUUCCUGGAUCAUGCCCGGAACCAUUAAGGAAAACAUCAUCUUCGGCGUGUCCUACGAUGAAUACCGCUACCGGUCCGUGAUCAAAGCCUGCCAGCUGGAAGAGGAUAUUUCAAAGUUCGCGGAGAAAGAUAACAUCGUGCUGGGCGAAGGGGGUAUUACCUUGUCGGGGGGCCAGCGGGCUAGAAUCUCGCUGGCCAGAGCCGUGUAUAAGGACGCCGACCUGUAUCUCCUGGACUCCCCCUUCGGAUACCUGGACGUCCUGACCGAAAAGGAGAUCUUCGAAUCGUGCGUGUGCAAGCUGAUGGCUAACAAGACUCGCAUCCUCGUGACCUCCAAAAUGGAGCACCUGAAGAAGGCAGACAAGAUUCUGAUUCUGCAUGAGGGGUCCUCCUACUUUUACGGCACCUUCUCGGAGUUGCAGAACUUGCAGCCCGACUUCUCAUCGAAGCUGAUGGGUUGCGACAGCUUCGACCAGUUCUCCGCCGAAAGAAGGAACUCGAUCCUGACGGAAACCUUGCACCGCUUCUCUUUGGAAGGCGACGCCCCUGUGUCAUGGACCGAGACUAAGAAGCAGAGCUUCAAGCAGACCGGGGAAUUCGGCGAAAAGAGGAAGAACAGCAUCUUGAACCCCAUUAACUCCAUCCGCAAGUUCUCAAUCGUGCAAAAGACGCCACUGCAGAUGAACGGCAUUGAGGAGGACUCCGACGAACCCCUUGAGAGGCGCCUGUCCCUGGUGCCGGACAGCGAGCAGGGAGAAGCCAUCCUGCCUCGGAUUUCCGUGAUCUCCACUGGUCCGACGCUCCAAGCCCGGCGGCGGCAGUCCGUGCUGAACCUGAUGACCCACAGCGUGAACCAGGGCCAAAACAUUCACCGCAAGACUACCGCAUCCACCCGGAAAGUGUCCCUGGCACCUCAAGCGAAUCUUACCGAGCUCGACAUCUACUCCCGGAGACUGUCGCAGGAAACCGGGCUCGAAAUUUCCGAAGAAAUCAACGAGGAGGAUCUGAAAGAGUGCUUCUUCGACGAUAUGGAGUCGAUACCCGCCGUGACGACUUGGAACACUUAUCUGCGGUACAUCACUGUGCACAAGUCAUUGAUCUUCGUGCUGAUUUGGUGCCUGGUGAUUUUCCUGGCCGAGGUCGCGGCCUCACUGGUGGUGCUCUGGCUGUUGGGAAACACGCCUCUGCAAGACAAGGGAAACUCCACGCACUCGAGAAACAACAGCUAUGCCGUGAUUAUCACUUCCACCUCCUCUUAUUACGUGUUCUACAUCUACGUCGGAGUGGCGGAUACCCUGCUCGCGAUGGGUUUCUUCAGAGGACUGCCGCUGGUCCACACCUUGAUCACCGUCAGCAAGAUUCUUCACCACAAGAUGUUGCAUAGCGUGCUGCAGGCCCCCAUGUCCACCCUCAACACUCUGAAGGCCGGAGGCAUUCUGAACAGAUUCUCCAAGGACAUCGCUAUCCUGGACGAUCUCCUGCCGCUUACCAUCUUUGACUUCAUCCAGCUGCUGCUGAUCGUGAUUGGAGCAAUCGCAGUGGUGGCGGUGCUGCAGCCUUACAUUUUCGUGGCCACUGUGCCGGUCAUUGUGGCGUUCAUCAUGCUGCGGGCCUACUUCCUCCAAACCAGCCAGCAGCUGAAGCAACUGGAAUCCGAGGGACGAUCCCCCAUCUUCACUCACCUUGUGACGUCGUUGAAGGGACUGUGGACCCUCCGGGCUUUCGGACGGCAGCCCUACUUCGAAACCCUCUUCCACAAGGCCCUGAACCUCCACACCGCCAAUUGGUUCCUGUACCUGUCCACCCUGCGGUGGUUCCAGAUGCGCAUCGAGAUGAUUUUCGUCAUCUUCUUCAUCGCGGUCACAUUCAUCAGCAUCCUGACUACCGGAGAGGGAGAGGGACGGGUCGGAAUAAUCCUGACCCUCGCCAUGAACAUUAUGAGCACCCUGCAGUGGGCAGUGAACAGCUCGAUCGACGUGGACAGCCUGAUGCGAAGCGUCAGCCGCGUGUUCAAGUUCAUCGACAUGCCUACUGAGGGAAAACCCACUAAGUCCACUAAGCCCUACAAAAAUGGCCAGCUGAGCAAGGUCAUGAUCAUCGAAAACUCCCACGUGAAGAAGGACGAUAUUUGGCCCUCCGGAGGUCAAAUGACCGUGAAGGACCUGACCGCAAAGUACACCGAGGGAGGAAACGCCAUUCUCGAAAACAUCAGCUUCUCCAUUUCGCCGGGACAGCGGGUCGGCCUUCUCGGGCGGACCGGUUCCGGGAAGUCAACUCUGCUGUCGGCUUUCCUCCGGCUGCUGAAUACCGAGGGGGAAAUCCAAAUUGACGGCGUGUCUUGGGAUUCCAUUACUCUGCAGCAGUGGCGGAAGGCCUUCGGCGUGAUCCCCCAGAAGGUGUUCAUCUUCUCGGGUACCUUCCGGAAGAACCUGGAUCCUUACGAGCAGUGGAGCGACCAAGAAAUCUGGAAGGUCGCCGACGAGGUCGGCCUGCGCUCCGUGAUUGAACAAUUUCCUGGAAAGCUGGACUUCGUGCUCGUCGACGGGGGAUGUGUCCUGUCGCACGGACAUAAGCAGCUCAUGUGCCUCGCACGGUCCGUGCUCUCCAAGGCCAAGAUUCUGCUGCUGGACGAACCUUCGGCCCACCUGGAUCCGGUCACCUACCAGAUCAUCAGGAGGACCCUGAAGCAGGCCUUUGCCGAUUGCACCGUGAUUCUCUGCGAGCACCGCAUCGAGGCCAUGCUGGAGUGCCAGCAGUUCCUGGUCAUCGAGGAGAACAAGGUCCGCCAAUACGACUCCAUUCAAAAGCUCCUCAACGAGCGGUCGCUGUUCAGACAAGCUAUUUCACCGUCCGAUAGAGUGAAGCUCUUCCCGCAUCGGAACAGCUCAAAGUGCAAAUCGAAGCCGCAGAUCGCAGCCUUGAAGGAAGAGACUGAGGAAGAGGUGCAGGACACCCGGCUUUAACGGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAUCAAGCU(SEQ ID NO:8)
在一个实施例中,另一个全长密码子优化的人CFTR mRNA序列为:
GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAGACACCGGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGCGGAUUCCCCGUGCCAAGAGUGACUCACCGUCCUUGACACGAUGCAACGCUCUCCUCUUGAAAAGGCCUCGGUGGUGUCCAAGCUCUUCUUCUCGUGGACUAGACCCAUCCUGAGAAAGGGGUACAGACAGCGCUUGGAGCUGUCCGAUAUCUAUCAAAUCCCUUCCGUGGACUCCGCGGACAACCUGUCCGAGAAGCUCGAGAGAGAAUGGGACAGAGAACUCGCCUCAAAGAAGAACCCGAAGCUGAUUAAUGCGCUUAGGCGGUGCUUUUUCUGGCGGUUCAUGUUCUACGGCAUCUUCCUCUACCUGGGAGAGGUCACCAAGGCCGUGCAGCCCCUGUUGCUGGGACGGAUUAUUGCCUCCUACGACCCCGACAACAAGGAAGAAAGAAGCAUCGCUAUCUACUUGGGCAUCGGUCUGUGCCUGCUUUUCAUCGUCCGGACCCUCUUGUUGCAUCCUGCUAUUUUCGGCCUGCAUCACAUUGGCAUGCAGAUGAGAAUUGCCAUGUUUUCCCUGAUCUACAAGAAAACUCUGAAGCUCUCGAGCCGCGUGCUUGACAAGAUUUCCAUCGGCCAGCUCGUGUCCCUGCUCUCCAACAAUCUGAACAAGUUCGACGAGGGCCUCGCCCUGGCCCACUUCGUGUGGAUCGCCCCUCUGCAAGUGGCGCUUCUGAUGGGCCUGAUCUGGGAGCUGCUGCAAGCCUCGGCAUUCUGUGGGCUUGGAUUCCUGAUCGUGCUGGCACUGUUCCAGGCCGGACUGGGGCGGAUGAUGAUGAAGUACAGGGACCAGAGAGCCGGAAAGAUUUCCGAACGGCUGGUGAUCACUUCGGAAAUGAUCGAAAACAUCCAGUCAGUGAAGGCCUACUGCUGGGAAGAGGCCAUGGAAAAGAUGAUUGAAAACCUCCGGCAAACCGAGCUGAAGCUGACCCGCAAGGCCGCUUACGUGCGCUAUUUCAACUCGUCCGCUUUCUUCUUCUCCGGGUUCUUCGUGGUGUUUCUCUCCGUGCUCCCCUACGCCCUGAUUAAGGGAAUCAUCCUCAGGAAGAUCUUCACCACCAUUUCCUUCUGUAUCGUGCUCCGCAUGGCCGUGACCCGGCAGUUCCCAUGGGCCGUGCAGACUUGGUACGACUCCCUGGGAGCCAUUAACAAGAUCCAGGACUUCCUUCAAAAGCAGGAGUACAAGACCCUCGAGUACAACCUGACUACUACCGAGGUCGUGAUGGAAAACGUCACCGCCUUUUGGGAGGAGGGAUUUGGCGAACUGUUCGAGAAGGCCAAGCAGAACAACAACAACCGCAAGACCUCGAACGGUGACGACUCCCUCUUCUUUUCAAACUUCAGCCUGCUCGGGACGCCCGUGCUGAAGGACAUUAACUUCAAGAUCGAAAGAGGACAGCUCCUGGCGGUGGCCGGAUCGACCGGAGCCGGAAAGACUUCCCUGCUGAUGGUGAUCAUGGGAGAGCUUGAACCUAGCGAGGGAAAGAUCAAGCACUCCGGCCGCAUCAGCUUCUGUAGCCAGUUUUCCUGGAUCAUGCCCGGAACCAUUAAGGAAAACAUCAUCUUCGGCGUGUCCUACGAUGAAUACCGCUACCGGUCCGUGAUCAAAGCCUGCCAGCUGGAAGAGGAUAUUUCAAAGUUCGCGGAGAAAGAUAACAUCGUGCUGGGCGAAGGGGGUAUUACCUUGUCGGGGGGCCAGCGGGCUAGAAUCUCGCUGGCCAGAGCCGUGUAUAAGGACGCCGACCUGUAUCUCCUGGACUCCCCCUUCGGAUACCUGGACGUCCUGACCGAAAAGGAGAUCUUCGAAUCGUGCGUGUGCAAGCUGAUGGCUAACAAGACUCGCAUCCUCGUGACCUCCAAAAUGGAGCACCUGAAGAAGGCAGACAAGAUUCUGAUUCUGCAUGAGGGGUCCUCCUACUUUUACGGCACCUUCUCGGAGUUGCAGAACUUGCAGCCCGACUUCUCAUCGAAGCUGAUGGGUUGCGACAGCUUCGACCAGUUCUCCGCCGAAAGAAGGAACUCGAUCCUGACGGAAACCUUGCACCGCUUCUCUUUGGAAGGCGACGCCCCUGUGUCAUGGACCGAGACUAAGAAGCAGAGCUUCAAGCAGACCGGGGAAUUCGGCGAAAAGAGGAAGAACAGCAUCUUGAACCCCAUUAACUCCAUCCGCAAGUUCUCAAUCGUGCAAAAGACGCCACUGCAGAUGAACGGCAUUGAGGAGGACUCCGACGAACCCCUUGAGAGGCGCCUGUCCCUGGUGCCGGACAGCGAGCAGGGAGAAGCCAUCCUGCCUCGGAUUUCCGUGAUCUCCACUGGUCCGACGCUCCAAGCCCGGCGGCGGCAGUCCGUGCUGAACCUGAUGACCCACAGCGUGAACCAGGGCCAAAACAUUCACCGCAAGACUACCGCAUCCACCCGGAAAGUGUCCCUGGCACCUCAAGCGAAUCUUACCGAGCUCGACAUCUACUCCCGGAGACUGUCGCAGGAAACCGGGCUCGAAAUUUCCGAAGAAAUCAACGAGGAGGAUCUGAAAGAGUGCUUCUUCGACGAUAUGGAGUCGAUACCCGCCGUGACGACUUGGAACACUUAUCUGCGGUACAUCACUGUGCACAAGUCAUUGAUCUUCGUGCUGAUUUGGUGCCUGGUGAUUUUCCUGGCCGAGGUCGCGGCCUCACUGGUGGUGCUCUGGCUGUUGGGAAACACGCCUCUGCAAGACAAGGGAAACUCCACGCACUCGAGAAACAACAGCUAUGCCGUGAUUAUCACUUCCACCUCCUCUUAUUACGUGUUCUACAUCUACGUCGGAGUGGCGGAUACCCUGCUCGCGAUGGGUUUCUUCAGAGGACUGCCGCUGGUCCACACCUUGAUCACCGUCAGCAAGAUUCUUCACCACAAGAUGUUGCAUAGCGUGCUGCAGGCCCCCAUGUCCACCCUCAACACUCUGAAGGCCGGAGGCAUUCUGAACAGAUUCUCCAAGGACAUCGCUAUCCUGGACGAUCUCCUGCCGCUUACCAUCUUUGACUUCAUCCAGCUGCUGCUGAUCGUGAUUGGAGCAAUCGCAGUGGUGGCGGUGCUGCAGCCUUACAUUUUCGUGGCCACUGUGCCGGUCAUUGUGGCGUUCAUCAUGCUGCGGGCCUACUUCCUCCAAACCAGCCAGCAGCUGAAGCAACUGGAAUCCGAGGGACGAUCCCCCAUCUUCACUCACCUUGUGACGUCGUUGAAGGGACUGUGGACCCUCCGGGCUUUCGGACGGCAGCCCUACUUCGAAACCCUCUUCCACAAGGCCCUGAACCUCCACACCGCCAAUUGGUUCCUGUACCUGUCCACCCUGCGGUGGUUCCAGAUGCGCAUCGAGAUGAUUUUCGUCAUCUUCUUCAUCGCGGUCACAUUCAUCAGCAUCCUGACUACCGGAGAGGGAGAGGGACGGGUCGGAAUAAUCCUGACCCUCGCCAUGAACAUUAUGAGCACCCUGCAGUGGGCAGUGAACAGCUCGAUCGACGUGGACAGCCUGAUGCGAAGCGUCAGCCGCGUGUUCAAGUUCAUCGACAUGCCUACUGAGGGAAAACCCACUAAGUCCACUAAGCCCUACAAAAAUGGCCAGCUGAGCAAGGUCAUGAUCAUCGAAAACUCCCACGUGAAGAAGGACGAUAUUUGGCCCUCCGGAGGUCAAAUGACCGUGAAGGACCUGACCGCAAAGUACACCGAGGGAGGAAACGCCAUUCUCGAAAACAUCAGCUUCUCCAUUUCGCCGGGACAGCGGGUCGGCCUUCUCGGGCGGACCGGUUCCGGGAAGUCAACUCUGCUGUCGGCUUUCCUCCGGCUGCUGAAUACCGAGGGGGAAAUCCAAAUUGACGGCGUGUCUUGGGAUUCCAUUACUCUGCAGCAGUGGCGGAAGGCCUUCGGCGUGAUCCCCCAGAAGGUGUUCAUCUUCUCGGGUACCUUCCGGAAGAACCUGGAUCCUUACGAGCAGUGGAGCGACCAAGAAAUCUGGAAGGUCGCCGACGAGGUCGGCCUGCGCUCCGUGAUUGAACAAUUUCCUGGAAAGCUGGACUUCGUGCUCGUCGACGGGGGAUGUGUCCUGUCGCACGGACAUAAGCAGCUCAUGUGCCUCGCACGGUCCGUGCUCUCCAAGGCCAAGAUUCUGCUGCUGGACGAACCUUCGGCCCACCUGGAUCCGGUCACCUACCAGAUCAUCAGGAGGACCCUGAAGCAGGCCUUUGCCGAUUGCACCGUGAUUCUCUGCGAGCACCGCAUCGAGGCCAUGCUGGAGUGCCAGCAGUUCCUGGUCAUCGAGGAGAACAAGGUCCGCCAAUACGACUCCAUUCAAAAGCUCCUCAACGAGCGGUCGCUGUUCAGACAAGCUAUUUCACCGUCCGAUAGAGUGAAGCUCUUCCCGCAUCGGAACAGCUCAAAGUGCAAAUCGAAGCCGCAGAUCGCAGCCUUGAAGGAAGAGACUGAGGAAGAGGUGCAGGACACCCGGCUUUAAGGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAUCAAAGCU(SEQ ID NO:9)
在一个实施例中,密码子优化的人CFTR mRNA序列包括SEQ ID NO:2。
在一个实施例中,全长密码子优化的人CFTR mRNA序列为:GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAGACACCGGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGCGGAUUCCCCGUGCCAAGAGUGACUCACCGUCCUUGACACGAUGCAGCGGUCCCCGCUCGAAAAGGCCAGUGUCGUGUCCAAACUCUUCUUCUCAUGGACUCGGCCUAUCCUUAGAAAGGGGUAUCGGCAGAGGCUUGAGUUGUCUGACAUCUACCAGAUCCCCUCGGUAGAUUCGGCGGAUAACCUCUCGGAGAAGCUCGAACGGGAAUGGGACCGCGAACUCGCGUCUAAGAAAAACCCGAAGCUCAUCAACGCACUGAGAAGGUGCUUCUUCUGGCGGUUCAUGUUCUACGGUAUCUUCUUGUAUCUCGGGGAGGUCACAAAAGCAGUCCAACCCCUGUUGUUGGGUCGCAUUAUCGCCUCGUACGACCCCGAUAACAAAGAAGAACGGAGCAUCGCGAUCUACCUCGGGAUCGGACUGUGUUUGCUUUUCAUCGUCAGAACACUUUUGUUGCAUCCAGCAAUCUUCGGCCUCCAUCACAUCGGUAUGCAGAUGCGAAUCGCUAUGUUUAGCUUGAUCUACAAAAAGACACUGAAACUCUCGUCGCGGGUGUUGGAUAAGAUUUCCAUCGGUCAGUUGGUGUCCCUGCUUAGUAAUAACCUCAACAAAUUCGAUGAGGGACUGGCGCUGGCACAUUUCGUGUGGAUUGCCCCGUUGCAAGUCGCCCUUUUGAUGGGCCUUAUUUGGGAGCUGUUGCAGGCAUCUGCCUUUUGUGGCCUGGGAUUUCUGAUUGUGUUGGCAUUGUUUCAGGCUGGGCUUGGGCGGAUGAUGAUGAAGUAUCGCGACCAGAGAGCGGGUAAAAUCUCGGAAAGACUCGUCAUCACUUCGGAAAUGAUCGAAAACAUCCAGUCGGUCAAAGCCUAUUGCUGGGAAGAAGCUAUGGAGAAGAUGAUUGAAAACCUCCGCCAAACUGAGCUGAAACUGACCCGCAAGGCGGCGUAUGUCCGGUAUUUCAAUUCGUCAGCGUUCUUCUUUUCCGGGUUCUUCGUUGUCUUUCUCUCGGUUUUGCCUUAUGCCUUGAUUAAGGGGAUUAUCCUCCGCAAGAUUUUCACCACGAUUUCGUUCUGCAUUGUAUUGCGCAUGGCAGUGACACGGCAAUUUCCGUGGGCCGUGCAGACAUGGUAUGACUCGCUUGGAGCGAUCAACAAAAUCCAAGACUUCUUGCAAAAGCAAGAGUACAAGACCCUGGAGUACAAUCUUACUACUACGGAGGUAGUAAUGGAGAAUGUGACGGCUUUUUGGGAAGAGGGUUUUGGAGAACUGUUUGAGAAAGCAAAGCAGAAUAACAACAACCGCAAGACCUCAAAUGGGGACGAUUCCCUGUUUUUCUCGAACUUCUCCCUGCUCGGAACACCCGUGUUGAAGGACAUCAAUUUCAAGAUUGAGAGGGGACAGCUUCUCGCGGUAGCGGGAAGCACUGGUGCGGGAAAAACUAGCCUCUUGAUGGUGAUUAUGGGGGAGCUUGAGCCCAGCGAGGGGAAGAUUAAACACUCCGGGCGUAUCUCAUUCUGUAGCCAGUUUUCAUGGAUCAUGCCCGGAACCAUUAAAGAGAACAUCAUUUUCGGAGUAUCCUAUGAUGAGUACCGAUACAGAUCGGUCAUUAAGGCGUGCCAGUUGGAAGAGGACAUUUCUAAGUUCGCCGAGAAGGAUAACAUCGUCUUGGGAGAAGGGGGUAUUACAUUGUCGGGAGGGCAGCGAGCGCGGAUCAGCCUCGCGAGAGCGGUAUACAAAGAUGCAGAUUUGUAUCUGCUUGAUUCACCGUUUGGAUACCUCGACGUAUUGACAGAAAAAGAAAUCUUCGAGUCGUGCGUGUGUAAACUUAUGGCUAAUAAGACGAGAAUCCUGGUGACAUCAAAAAUGGAACACCUUAAGAAGGCGGACAAGAUCCUGAUCCUCCACGAAGGAUCGUCCUACUUUUACGGCACUUUCUCAGAGUUGCAAAACUUGCAGCCGGACUUCUCAAGCAAACUCAUGGGGUGUGACUCAUUCGACCAGUUCAGCGCGGAACGGCGGAACUCGAUCUUGACGGAAACGCUGCACCGAUUCUCGCUUGAGGGUGAUGCCCCGGUAUCGUGGACCGAGACAAAGAAGCAGUCGUUUAAGCAGACAGGAGAAUUUGGUGAGAAAAGAAAGAACAGUAUCUUGAAUCCUAUUAACUCAAUUCGCAAGUUCUCAAUCGUCCAGAAAACUCCACUGCAGAUGAAUGGAAUUGAAGAGGAUUCGGACGAACCCCUGGAGCGCAGGCUUAGCCUCGUGCCGGAUUCAGAGCAAGGGGAGGCCAUUCUUCCCCGGAUUUCGGUGAUUUCAACCGGACCUACACUUCAGGCGAGGCGAAGGCAAUCCGUGCUCAACCUCAUGACGCAUUCGGUAAACCAGGGGCAAAACAUUCACCGCAAAACGACGGCCUCAACGAGAAAAGUGUCACUUGCACCCCAGGCGAAUUUGACUGAACUCGACAUCUACAGCCGUAGGCUUUCGCAAGAAACCGGACUUGAGAUCAGCGAAGAAAUCAAUGAAGAAGAUUUGAAAGAGUGUUUCUUUGAUGACAUGGAAUCAAUCCCAGCGGUGACAACGUGGAACACAUACUUGCGUUACAUCACGGUGCACAAGUCCUUGAUUUUCGUCCUCAUCUGGUGUCUCGUGAUCUUUCUCGCUGAGGUCGCAGCGUCACUUGUGGUCCUCUGGCUGCUUGGUAAUACGCCCUUGCAAGACAAAGGCAAUUCUACACACUCAAGAAACAAUUCCUAUGCCGUGAUUAUCACUUCUACAAGCUCGUAUUACGUGUUUUACAUCUACGUAGGAGUGGCCGACACUCUGCUCGCGAUGGGUUUCUUCCGAGGACUCCCACUCGUUCACACGCUUAUCACUGUCUCCAAGAUUCUCCACCAUAAGAUGCUUCAUAGCGUACUGCAGGCUCCCAUGUCCACCUUGAAUACGCUCAAGGCGGGAGGUAUUUUGAAUCGCUUCUCAAAAGAUAUUGCAAUUUUGGAUGACCUUCUGCCCCUGACGAUCUUCGACUUCAUCCAGUUGUUGCUGAUCGUGAUUGGGGCUAUUGCAGUAGUCGCUGUCCUCCAGCCUUACAUUUUUGUCGCGACCGUUCCGGUGAUCGUGGCGUUUAUCAUGCUGCGGGCCUAUUUCUUGCAGACGUCACAGCAGCUUAAGCAACUGGAGUCUGAAGGGAGGUCGCCUAUCUUUACGCAUCUUGUGACCAGUUUGAAGGGAUUGUGGACGUUGCGCGCCUUUGGCAGGCAGCCCUACUUUGAAACACUGUUCCACAAAGCGCUGAAUCUCCAUACGGCAAAUUGGUUUUUGUAUUUGAGUACCCUCCGAUGGUUUCAGAUGCGCAUUGAGAUGAUUUUUGUGAUCUUCUUUAUCGCGGUGACUUUUAUCUCCAUCUUGACCACGGGAGAGGGCGAGGGACGGGUCGGUAUUAUCCUGACACUCGCCAUGAACAUUAUGAGCACUUUGCAGUGGGCAGUGAACAGCUCGAUUGAUGUGGAUAGCCUGAUGAGGUCCGUUUCGAGGGUCUUUAAGUUCAUCGACAUGCCGACGGAGGGAAAGCCCACAAAAAGUACGAAACCCUAUAAGAAUGGGCAAUUGAGUAAGGUAAUGAUCAUCGAGAACAGUCACGUGAAGAAGGAUGACAUCUGGCCUAGCGGGGGUCAGAUGACCGUGAAGGACCUGACGGCAAAAUACACCGAGGGAGGGAACGCAAUCCUUGAAAACAUCUCGUUCAGCAUUAGCCCCGGUCAGCGUGUGGGGUUGCUCGGGAGGACCGGGUCAGGAAAAUCGACGUUGCUGUCGGCCUUCUUGAGACUUCUGAAUACAGAGGGUGAGAUCCAGAUCGACGGCGUUUCGUGGGAUAGCAUCACCUUGCAGCAGUGGCGGAAAGCGUUUGGAGUAAUCCCCCAAAAGGUCUUUAUCUUUAGCGGAACCUUCCGAAAGAAUCUCGAUCCUUAUGAACAGUGGUCAGAUCAAGAGAUUUGGAAAGUCGCGGACGAGGUUGGCCUUCGGAGUGUAAUCGAGCAGUUUCCGGGAAAACUCGACUUUGUCCUUGUAGAUGGGGGAUGCGUCCUGUCGCAUGGGCACAAGCAGCUCAUGUGCCUGGCGCGAUCCGUCCUCUCUAAAGCGAAAAUUCUUCUCUUGGAUGAACCUUCGGCCCAUCUGGACCCGGUAACGUAUCAGAUCAUCAGAAGGACACUUAAGCAGGCGUUUGCCGACUGCACGGUGAUUCUCUGUGAGCAUCGUAUCGAGGCCAUGCUCGAAUGCCAGCAAUUUCUUGUCAUCGAAGAGAAUAAGGUCCGCCAGUACGACUCCAUCCAGAAGCUGCUUAAUGAGAGAUCAUUGUUCCGGCAGGCGAUUUCACCAUCCGAUAGGGUGAAACUUUUUCCACACAGAAAUUCGUCGAAGUGCAAGUCCAAACCGCAGAUCGCGGCCUUGAAAGAAGAGACUGAAGAAGAAGUUCAAGACACGCGUCUUUAACGGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAUCAAGCU(SEQ ID NO:10)3]
在一个实施例中,另一个全长密码子优化的人CFTR mRNA序列为:
GGACAGAUCGCCUGGAGACGCCAUCCACGCUGUUUUGACCUCCAUAGAAGACACCGGGACCGAUCCAGCCUCCGCGGCCGGGAACGGUGCAUUGGAACGCGGAUUCCCCGUGCCAAGAGUGACUCACCGUCCUUGACACGAUGCAGCGGUCCCCGCUCGAAAAGGCCAGUGUCGUGUCCAAACUCUUCUUCUCAUGGACUCGGCCUAUCCUUAGAAAGGGGUAUCGGCAGAGGCUUGAGUUGUCUGACAUCUACCAGAUCCCCUCGGUAGAUUCGGCGGAUAACCUCUCGGAGAAGCUCGAACGGGAAUGGGACCGCGAACUCGCGUCUAAGAAAAACCCGAAGCUCAUCAACGCACUGAGAAGGUGCUUCUUCUGGCGGUUCAUGUUCUACGGUAUCUUCUUGUAUCUCGGGGAGGUCACAAAAGCAGUCCAACCCCUGUUGUUGGGUCGCAUUAUCGCCUCGUACGACCCCGAUAACAAAGAAGAACGGAGCAUCGCGAUCUACCUCGGGAUCGGACUGUGUUUGCUUUUCAUCGUCAGAACACUUUUGUUGCAUCCAGCAAUCUUCGGCCUCCAUCACAUCGGUAUGCAGAUGCGAAUCGCUAUGUUUAGCUUGAUCUACAAAAAGACACUGAAACUCUCGUCGCGGGUGUUGGAUAAGAUUUCCAUCGGUCAGUUGGUGUCCCUGCUUAGUAAUAACCUCAACAAAUUCGAUGAGGGACUGGCGCUGGCACAUUUCGUGUGGAUUGCCCCGUUGCAAGUCGCCCUUUUGAUGGGCCUUAUUUGGGAGCUGUUGCAGGCAUCUGCCUUUUGUGGCCUGGGAUUUCUGAUUGUGUUGGCAUUGUUUCAGGCUGGGCUUGGGCGGAUGAUGAUGAAGUAUCGCGACCAGAGAGCGGGUAAAAUCUCGGAAAGACUCGUCAUCACUUCGGAAAUGAUCGAAAACAUCCAGUCGGUCAAAGCCUAUUGCUGGGAAGAAGCUAUGGAGAAGAUGAUUGAAAACCUCCGCCAAACUGAGCUGAAACUGACCCGCAAGGCGGCGUAUGUCCGGUAUUUCAAUUCGUCAGCGUUCUUCUUUUCCGGGUUCUUCGUUGUCUUUCUCUCGGUUUUGCCUUAUGCCUUGAUUAAGGGGAUUAUCCUCCGCAAGAUUUUCACCACGAUUUCGUUCUGCAUUGUAUUGCGCAUGGCAGUGACACGGCAAUUUCCGUGGGCCGUGCAGACAUGGUAUGACUCGCUUGGAGCGAUCAACAAAAUCCAAGACUUCUUGCAAAAGCAAGAGUACAAGACCCUGGAGUACAAUCUUACUACUACGGAGGUAGUAAUGGAGAAUGUGACGGCUUUUUGGGAAGAGGGUUUUGGAGAACUGUUUGAGAAAGCAAAGCAGAAUAACAACAACCGCAAGACCUCAAAUGGGGACGAUUCCCUGUUUUUCUCGAACUUCUCCCUGCUCGGAACACCCGUGUUGAAGGACAUCAAUUUCAAGAUUGAGAGGGGACAGCUUCUCGCGGUAGCGGGAAGCACUGGUGCGGGAAAAACUAGCCUCUUGAUGGUGAUUAUGGGGGAGCUUGAGCCCAGCGAGGGGAAGAUUAAACACUCCGGGCGUAUCUCAUUCUGUAGCCAGUUUUCAUGGAUCAUGCCCGGAACCAUUAAAGAGAACAUCAUUUUCGGAGUAUCCUAUGAUGAGUACCGAUACAGAUCGGUCAUUAAGGCGUGCCAGUUGGAAGAGGACAUUUCUAAGUUCGCCGAGAAGGAUAACAUCGUCUUGGGAGAAGGGGGUAUUACAUUGUCGGGAGGGCAGCGAGCGCGGAUCAGCCUCGCGAGAGCGGUAUACAAAGAUGCAGAUUUGUAUCUGCUUGAUUCACCGUUUGGAUACCUCGACGUAUUGACAGAAAAAGAAAUCUUCGAGUCGUGCGUGUGUAAACUUAUGGCUAAUAAGACGAGAAUCCUGGUGACAUCAAAAAUGGAACACCUUAAGAAGGCGGACAAGAUCCUGAUCCUCCACGAAGGAUCGUCCUACUUUUACGGCACUUUCUCAGAGUUGCAAAACUUGCAGCCGGACUUCUCAAGCAAACUCAUGGGGUGUGACUCAUUCGACCAGUUCAGCGCGGAACGGCGGAACUCGAUCUUGACGGAAACGCUGCACCGAUUCUCGCUUGAGGGUGAUGCCCCGGUAUCGUGGACCGAGACAAAGAAGCAGUCGUUUAAGCAGACAGGAGAAUUUGGUGAGAAAAGAAAGAACAGUAUCUUGAAUCCUAUUAACUCAAUUCGCAAGUUCUCAAUCGUCCAGAAAACUCCACUGCAGAUGAAUGGAAUUGAAGAGGAUUCGGACGAACCCCUGGAGCGCAGGCUUAGCCUCGUGCCGGAUUCAGAGCAAGGGGAGGCCAUUCUUCCCCGGAUUUCGGUGAUUUCAACCGGACCUACACUUCAGGCGAGGCGAAGGCAAUCCGUGCUCAACCUCAUGACGCAUUCGGUAAACCAGGGGCAAAACAUUCACCGCAAAACGACGGCCUCAACGAGAAAAGUGUCACUUGCACCCCAGGCGAAUUUGACUGAACUCGACAUCUACAGCCGUAGGCUUUCGCAAGAAACCGGACUUGAGAUCAGCGAAGAAAUCAAUGAAGAAGAUUUGAAAGAGUGUUUCUUUGAUGACAUGGAAUCAAUCCCAGCGGUGACAACGUGGAACACAUACUUGCGUUACAUCACGGUGCACAAGUCCUUGAUUUUCGUCCUCAUCUGGUGUCUCGUGAUCUUUCUCGCUGAGGUCGCAGCGUCACUUGUGGUCCUCUGGCUGCUUGGUAAUACGCCCUUGCAAGACAAAGGCAAUUCUACACACUCAAGAAACAAUUCCUAUGCCGUGAUUAUCACUUCUACAAGCUCGUAUUACGUGUUUUACAUCUACGUAGGAGUGGCCGACACUCUGCUCGCGAUGGGUUUCUUCCGAGGACUCCCACUCGUUCACACGCUUAUCACUGUCUCCAAGAUUCUCCACCAUAAGAUGCUUCAUAGCGUACUGCAGGCUCCCAUGUCCACCUUGAAUACGCUCAAGGCGGGAGGUAUUUUGAAUCGCUUCUCAAAAGAUAUUGCAAUUUUGGAUGACCUUCUGCCCCUGACGAUCUUCGACUUCAUCCAGUUGUUGCUGAUCGUGAUUGGGGCUAUUGCAGUAGUCGCUGUCCUCCAGCCUUACAUUUUUGUCGCGACCGUUCCGGUGAUCGUGGCGUUUAUCAUGCUGCGGGCCUAUUUCUUGCAGACGUCACAGCAGCUUAAGCAACUGGAGUCUGAAGGGAGGUCGCCUAUCUUUACGCAUCUUGUGACCAGUUUGAAGGGAUUGUGGACGUUGCGCGCCUUUGGCAGGCAGCCCUACUUUGAAACACUGUUCCACAAAGCGCUGAAUCUCCAUACGGCAAAUUGGUUUUUGUAUUUGAGUACCCUCCGAUGGUUUCAGAUGCGCAUUGAGAUGAUUUUUGUGAUCUUCUUUAUCGCGGUGACUUUUAUCUCCAUCUUGACCACGGGAGAGGGCGAGGGACGGGUCGGUAUUAUCCUGACACUCGCCAUGAACAUUAUGAGCACUUUGCAGUGGGCAGUGAACAGCUCGAUUGAUGUGGAUAGCCUGAUGAGGUCCGUUUCGAGGGUCUUUAAGUUCAUCGACAUGCCGACGGAGGGAAAGCCCACAAAAAGUACGAAACCCUAUAAGAAUGGGCAAUUGAGUAAGGUAAUGAUCAUCGAGAACAGUCACGUGAAGAAGGAUGACAUCUGGCCUAGCGGGGGUCAGAUGACCGUGAAGGACCUGACGGCAAAAUACACCGAGGGAGGGAACGCAAUCCUUGAAAACAUCUCGUUCAGCAUUAGCCCCGGUCAGCGUGUGGGGUUGCUCGGGAGGACCGGGUCAGGAAAAUCGACGUUGCUGUCGGCCUUCUUGAGACUUCUGAAUACAGAGGGUGAGAUCCAGAUCGACGGCGUUUCGUGGGAUAGCAUCACCUUGCAGCAGUGGCGGAAAGCGUUUGGAGUAAUCCCCCAAAAGGUCUUUAUCUUUAGCGGAACCUUCCGAAAGAAUCUCGAUCCUUAUGAACAGUGGUCAGAUCAAGAGAUUUGGAAAGUCGCGGACGAGGUUGGCCUUCGGAGUGUAAUCGAGCAGUUUCCGGGAAAACUCGACUUUGUCCUUGUAGAUGGGGGAUGCGUCCUGUCGCAUGGGCACAAGCAGCUCAUGUGCCUGGCGCGAUCCGUCCUCUCUAAAGCGAAAAUUCUUCUCUUGGAUGAACCUUCGGCCCAUCUGGACCCGGUAACGUAUCAGAUCAUCAGAAGGACACUUAAGCAGGCGUUUGCCGACUGCACGGUGAUUCUCUGUGAGCAUCGUAUCGAGGCCAUGCUCGAAUGCCAGCAAUUUCUUGUCAUCGAAGAGAAUAAGGUCCGCCAGUACGACUCCAUCCAGAAGCUGCUUAAUGAGAGAUCAUUGUUCCGGCAGGCGAUUUCACCAUCCGAUAGGGUGAAACUUUUUCCACACAGAAAUUCGUCGAAGUGCAAGUCCAAACCGCAGAUCGCGGCCUUGAAAGAAGAGACUGAAGAAGAAGUUCAAGACACGCGUCUUUAAGGGUGGCAUCCCUGUGACCCCUCCCCAGUGCCUCUCCUGGCCCUGGAAGUUGCCACUCCAGUGCCCACCAGCCUUGUCCUAAUAAAAUUAAGUUGCAUCAAAGCU(SEQ ID NO:11)
SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10和SEQ ID NO:11包括为密码子优化的hCFTR编码mRNA制定框架的5'和3'非翻译区。
在一些实施例中,合适的mRNA序列可为人CFTR(hCFTR)蛋白的同源物或类似物的mRNA序列。例如,hCFTR蛋白的同源物或类似物可为经修饰的hCFTR蛋白,与野生型或天然存在的hCFTR蛋白相比,其含有一个或多个氨基酸取代、缺失和/或插入,同时保留基本的hCFTR蛋白活性。在一些实施例中,适合于本发明的mRNA编码与SEQ ID NO:4具有至少50%、55%、60%、65%、70%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或更大同源性的氨基酸序列。在一些实施例中,适合于本发明的mRNA编码与hCFTR蛋白基本上相同的蛋白质。在一些实施例中,适合于本发明的mRNA编码与SEQ ID NO:4具有至少50%、55%、60%、65%、70%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或更大一致性的氨基酸序列。在一些实施例中,适合于本发明的mRNA编码hCFTR蛋白的片段或部分。在一些实施例中,适合于本发明的mRNA编码hCFTR蛋白的片段或部分,其中所述蛋白的片段或部分仍保持与野生型蛋白类似的CFTR活性。在一些实施例中,适合于本发明的mRNA具有与SEQ ID NO:1、SEQ ID NO:8或SEQ ID NO:9具有至少50%、55%、60%、65%、70%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或更大一致性的核苷酸序列。在一些实施例中,适合于本发明的mRNA具有与SEQ ID NO:2、SEQ ID NO:10或SEQ ID NO:11具有至少50%、55%、60%、65%、70%、75%、80%、85%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或更大一致性的核苷酸序列。在一些实施例中,适合于本发明的mRNA包含与SEQ ID NO:1相同的核苷酸序列。在一些实施例中,适合于本发明的mRNA包含与SEQ ID NO:8相同的核苷酸序列。在一些实施例中,适合于本发明的mRNA包含与SEQ ID NO:9相同的核苷酸序列。在一些实施例中,合适的mRNA编码融合蛋白,所述融合蛋白包含与另一种蛋白质融合的hCFTR蛋白的全长、片段或部分(例如,N端或C端融合物)在一些实施例中,与编码hCFTR蛋白的全长、片段或部分的mRNA融合的蛋白质编码信号序列或细胞靶向序列。
可根据各种已知方法中的任一种合成根据本发明的mRNA。例如,可经由体外转录(IVT)合成根据本发明的mRNA。简言之,IVT通常用含有启动子的线性或环状DNA模板、核糖核苷酸三磷酸库、可包括DTT和镁离子的缓冲系统、以及适当的RNA聚合酶(例如T3、T7或SP6RNA聚合酶)、DNA酶I、焦磷酸酶和/或RNA酶抑制剂执行。确切条件将根据具体应用而改变。
非编码区
在一些实施例中,mRNA包括5'和/或3'非翻译区。在一些实施例中,5'非翻译区包括影响mRNA的稳定性或翻译的一种或多种元件,例如铁应答元件。在一些实施例中,5'非翻译区的长度可为约50至500个核苷酸。
在一些实施例中,3'非翻译区包括聚腺苷酸化信号、影响mRNA在细胞中的定位稳定性的蛋白质的结合位点、或miRNA的一个或多个结合位点中的一种或多种。在一些实施例中,3'非翻译区的长度可为50至500个核苷酸或更长。
示例性的3'和/或5'UTR序列可源自稳定的mRNA分子(例如珠蛋白、肌动蛋白、GAPDH、微管蛋白、组蛋白或柠檬酸循环酶),以增加感觉mRNA分子的稳定性。例如,5'UTR序列可包括CMV立即早期1(IE1)基因的部分序列或其片段,以改善核酸酶抗性和/或改善多核苷酸的半衰期。还考虑了将编码人生长激素(hGH)或其片段的序列包括在多核苷酸(例如mRNA)的3'端或非翻译区,以进一步稳定多核苷酸。一般地,这些修饰相对于其未修饰的对应物改善了多核苷酸的稳定性和/或药代动力学特性(例如半衰期),并且包括例如为改善此类多核苷酸对体内核酸酶消化的抗性而进行的修饰。
根据各种实施例,任何尺寸mRNA可通过提供的脂质体来囊封。在一些实施例中,提供的脂质体可囊封长度超过约0.5kb、1kb、1.5kb、2kb、2.5kb、3kb、3.5kb、4kb、4.5kb或5kb的mRNA。
通常,mRNA合成包括在N端(5')末端上添加“帽”,和在C端(3')末端上添加“尾”。帽的存在对于提供针对大部分真核细胞中发现的核酸酶的抗性是重要的。“尾”的存在用以保护mRNA免于核酸外切酶降解。
因此,在一些实施例中,mRNA(例如编码CFTR的mRNA)包括5'帽结构。5'帽通常如下添加:首先,RNA末端磷酸酶从5'核苷酸中去除一个末端磷酸基,留下两个末端磷酸盐;然后经由鸟苷酸转移酶将鸟苷三磷酸(GTP)加入末端磷酸盐中,产生5'5'5三磷酸键;且接着通过甲基转移酶将鸟嘌呤的7-氮甲基化。帽结构的实例包括但不限于m7G(5')ppp(5'(A,G(5')ppp(5')A和G(5')ppp(5')G。额外的帽结构描述于公布的美国申请第US 2016/0032356号和2017年2月27日提交的美国临时申请62/464,327中,其以引用的方式并入本文中。
在一些实施例中,帽是Cap0结构。Cap0结构缺少与碱基1和2连接的核糖的2'-O-甲基残基。在一些实施例中,帽是Cap1结构。Cap1结构在碱基2处具有2'-O-甲基残基。在一些实施例中,帽是Cap2结构。Cap2结构具有与碱基2和3两者连接的2'-O-甲基残基。
在一些实施例中,mRNA(例如编码CFTR的mRNA)包括3'尾结构。通常,尾结构包括poly(A)和/或poly(C)尾。mRNA的3'末端上的poly-A或poly-C尾通常分别包括至少50个腺苷或胞嘧啶核苷酸、至少150个腺苷或胞嘧啶核苷酸、至少200个腺苷或胞嘧啶核苷酸、至少250个腺苷或胞嘧啶核苷酸、至少300个腺苷或胞嘧啶核苷酸、至少350个腺苷或胞嘧啶核苷酸、至少400个腺苷或胞嘧啶核苷酸、至少450个腺苷或胞嘧啶核苷酸、至少500个腺苷或胞嘧啶核苷酸、至少550个腺苷或胞嘧啶核苷酸、至少600个腺苷或胞嘧啶核苷酸、至少650个腺苷酸或胞嘧啶核苷酸、至少700个腺苷或胞嘧啶核苷酸、至少750个腺苷或胞嘧啶核苷酸、至少800个腺苷或胞嘧啶核苷酸、至少850个腺苷或胞嘧啶核苷酸、至少900个腺苷或胞嘧啶核苷酸、至少950个腺苷或胞嘧啶核苷酸或至少1kb个腺苷或胞嘧啶核苷酸。在一些实施例中,poly-A或poly-C尾可分别为约10至800个腺苷或胞嘧啶核苷酸(例如约10至200个腺苷或胞嘧啶核苷酸、约10至300个腺苷或胞嘧啶核苷酸、约10至400个腺苷或胞嘧啶核苷酸、约10至500个腺苷或胞嘧啶核苷酸、约10至550个腺苷或胞嘧啶核苷酸、约10至600个腺苷或胞嘧啶核苷酸、约50至600个腺苷或胞嘧啶核苷酸、约100至600个腺苷或胞嘧啶核苷酸、约150至600个腺苷或胞嘧啶核苷酸、约200至600个腺苷或胞嘧啶核苷酸、约250至600个腺苷或胞嘧啶核苷酸、约300至600个腺苷或胞嘧啶核苷酸、约350至600个腺苷或胞嘧啶核苷酸、约400至600个腺苷或胞嘧啶核苷酸、约450至600个腺苷或胞嘧啶核苷酸、约500至600个腺苷或胞嘧啶核苷酸、约10至150个腺苷或胞嘧啶核苷酸、约10至100个腺苷或胞嘧啶核苷酸、约20至70个腺苷或胞嘧啶核苷酸或约20至60个腺苷或胞嘧啶核苷酸)。在一些实施例中,尾结构包括具有本文所述的各种长度的poly(A)和poly(C)尾的组合。在一些实施例中,尾结构包括至少50%、55%、65%、70%、75%、80%、85%、90%、92%、94%、95%、96%、97%、98%或99%的腺苷核苷酸。在一些实施例中,尾结构包括至少50%、55%、65%、70%、75%、80%、85%、90%、92%、94%、95%、96%、97%、98%或99%的胞嘧啶核苷酸。
经修饰的mRNA
在一些实施例中,根据本发明的mRNA可合成为未经修饰或经修饰的mRNA。通常,修饰mRNA以增强稳定性。mRNA的修饰可包括例如mRNA的核苷酸的修饰。因此,根据本发明的经修饰的mRNA可包括例如主链修饰、糖修饰或碱基修饰。在一些实施例中,mRNA可由天然存在的核苷酸和/或核苷酸类似物(经修饰的核苷酸)合成,包括但不限于嘌呤(腺嘌呤(A)、鸟嘌呤(G))或嘧啶(胸腺嘧啶(T)、胞嘧啶(C)、尿嘧啶(U)),以及作为嘌呤和嘧啶的经修饰的核苷酸类似物或衍生物,例如1-甲基-腺嘌呤、2-甲基-腺嘌呤、2-甲硫基-N-6-异戊烯基-腺嘌呤、N6-甲基-腺嘌呤、N6-异戊烯基-腺嘌呤、2-硫代-胞嘧啶、3-甲基-胞嘧啶、4-乙酰基-胞嘧啶、5-甲基-胞嘧啶、2,6-二氨基嘌呤、1-甲基-鸟嘌呤、2-甲基-鸟嘌呤、2,2-二甲基-鸟嘌呤、7-甲基-鸟嘌呤、肌苷、1-甲基肌苷、假尿嘧啶(5-尿嘧啶)、二氢尿嘧啶、2-硫代尿嘧啶、4-硫代尿嘧啶、5-羧甲基氨基甲基-2-硫代尿嘧啶、5-(羧基羟甲基)-尿嘧啶、5-氟尿嘧啶、5-溴尿嘧啶、5-羧甲基氨基甲基-尿嘧啶、5-甲基-2-硫代尿嘧啶、5-甲基尿嘧啶、N-尿嘧啶-5-氧乙酸甲酯、5-甲基氨基甲基-尿嘧啶、5-甲氧基氨基甲基-2-硫代尿嘧啶、5'-甲氧基羰基甲基-尿嘧啶、5-甲氧基-尿嘧啶、尿嘧啶-5-氧乙酸甲酯、尿嘧啶-5-氧乙酸(v)、1-甲基-假尿嘧啶、Q核苷、β-D-甘露糖基-Q核苷、怀丁苷(wybutoxosine)和氨基磷酸酯、硫代磷酸酯、肽核苷酸、甲基膦酸酯、7-脱氮鸟苷、5-甲基胞嘧啶和肌苷。这些类似物的制备是所属领域的技术人员已知的,例如根据美国专利第4,373,071号、美国专利第4,401,796号、美国专利第4,415,732号、美国专利第4,458,066号、美国专利第4,500,707号、美国专利第4,668,777号、美国专利第4,973,679号、美国专利第5,047,524号、美国专利第5,132,418号、美国专利第5,153,319号、美国专利第5,262,530号和第5,700,642号,其公开内容以全文引用的方式并入。
在一些实施例中,mRNA(例如编码CFTR的mRNA)可含有RNA主链修饰。通常,主链修饰是其中包含在RNA中的核苷酸主链的磷酸酯经化学修饰的修饰。示例性的主链修饰通常包括但不限于来自由以下组成的群组的修饰:甲基膦酸酯、甲基氨基磷酸酯、氨基磷酸酯、硫代磷酸酯(例如胞苷5'-O-(1-硫代磷酸酯))、硼烷磷酸酯、带正电的胍基团等,其意指通过其它阴离子、阳离子或中性基团置换磷酸二酯键。
在一些实施例中,mRNA(例如编码CFTR的mRNA)可含有糖修饰。典型的糖修饰是其所含有的核苷酸的糖的化学修饰,包括但不限于选自由以下组成的群组的糖修饰:2'-脱氧-2'-氟-寡核糖核苷酸(2'-氟-2'-脱氧胞苷5'-三磷酸、2'-氟-2'-脱氧尿苷5'-三磷酸)、2'-脱氧-2'-脱胺-寡核糖核苷酸(2'-氨基-2'-脱氧胞苷5'-三磷酸、2'-氨基-2'-脱氧尿苷5'-三磷酸)、2'-O-烷基寡核糖核苷酸、2'-脱氧-2'-C-烷基寡核糖核苷酸(2'-O-甲基胞苷5'-三磷酸、2'-甲基尿苷5'-三磷酸)、2'-C-烷基寡核糖核苷酸和其异构体(2'-阿糖胞苷5'-三磷酸、2'-阿糖尿苷5'-三磷酸)或叠氮三磷酸(2'-叠氮基-2'-脱氧胞苷5'-三磷酸、2'-叠氮基-2'-脱氧尿苷5'-三磷酸)或4'-硫基取代的核糖核苷酸。
在一些实施例中,mRNA(例如编码CFTR的mRNA)可含有核苷酸碱基的修饰(碱基修饰)。含有碱基修饰的经修饰的核苷酸也称为碱基修饰的核苷酸。此类碱基修饰的核苷酸的例子包括但不限于2-氨基-6-氯嘌呤核苷5'-三磷酸、2-氨基腺苷5'-三磷酸、2-硫代胞苷5'-三磷酸、2-硫代尿苷5'-三磷酸、4-硫代尿苷5'-三磷酸、5-氨基烯丙基胞苷5'-三磷酸、5-氨基烯丙基尿苷5'-三磷酸、5-溴胞苷5'-三磷酸、5-溴尿苷5'-三磷酸、5-碘胞苷5'-三磷酸、5-碘尿苷5'-三磷酸、5-甲基胞苷5'-三磷酸、5-甲基尿苷5'-三磷酸、6-氮杂胞苷5'-三磷酸、6-氮杂尿苷5'-三磷酸、6-氯嘌呤核苷5'-三磷酸、7-脱氮腺苷5'-三磷酸、7-脱氮鸟苷5'-三磷酸、8-氮杂腺苷5'-三磷酸、8-叠氮基腺苷5'-三磷酸、苯并咪唑核苷5'-三磷酸、N1-甲基腺苷5'-三磷酸、N1-甲基鸟苷5'-三磷酸、N6-甲基腺苷5'-三磷酸、O6-甲基鸟苷5'-三磷酸、假尿苷5'-三磷酸、嘌呤霉素5'-三磷酸或黄苷5'-三磷酸。
药物组合物
为了促进mRNA在体内的表达,可将例如脂质体的递送媒剂与一种或多种另外的核酸、载体、靶向配体或稳定剂组合,或在药理学组合物中配制,在所述药理学组合物中其与合适的赋形剂混合。用于配制和施用药物的技术可在“雷明顿药物科学(Remington'sPharmaceutical Sciences)”,Mack Publishing Co.,Easton,Pa.,最新版中找到。
所提供的脂质体囊封的或结合的mRNA,以及含有其的组合物,可根据当前的医学实践施用和给药,考虑到个体的临床状况、施用部位和施用方法、施用的时间安排、个体的年龄、性别、体重以及与具有所属领域的一般技术的临床医生相关的其它因素。用于本文目的的“有效量”可通过如实验临床研究、药理学、临床和医学领域的普通技术人员已知的此类相关考虑因素来确定。在一些实施例中,所施用的量有效地实现如所属领域的技术人员选择作为疾病进展、消退或改善的适当量度的症状和其它指标的至少一些稳定、改善或消除。例如,合适的量和给药方案是引起至少瞬时蛋白质(例如酶)产生的那种。
合适的施用途径包括例如经口、经直肠、经阴道、经粘膜、经肺(包括气管内或吸入)、或肠道施用;肠胃外递送,包括皮内、经皮(局部)、肌肉内、皮下、髓内注射,以及鞘内、直接心室内、静脉内、腹膜内和/或鼻内施用。
替代地或另外,本发明的脂质体囊封的mRNA和组合物可以局部而不是全身方式施用,例如,经由将药物组合物直接注射到靶向组织内,优选以持续释放制剂。取决于待靶向的组织,局部递送可以各种方式受到影响。例如,可吸入含有本发明的组合物的气溶胶(用于鼻、气管或支气管递送);例如,可将本发明的组合物注射到损伤、疾病表现或疼痛部位内;组合物可在用于经口、气管或食道应用的锭剂中提供;可以用于施用于胃或肠的液体、片剂或胶囊形式供应,可以用于直肠或阴道应用的栓剂形式供应;或者甚至可通过使用乳膏、滴剂或甚至注射剂递送到眼。含有与治疗分子或配体复合的所提供组合物的制剂甚至可以手术方式施用,例如与可允许组合物从植入部位扩散到周围细胞的聚合物或其它结构或物质结合。或者,其可以手术方式施用,而无需使用聚合物或载体。
在一些实施例中,提供的脂质体和/或组合物被配制成使其适合于其中包含的mRNA的延长释放。此类延长释放组合物可以延长的给药间隔方便地施用于个体。例如,在一个实施例中,本发明的组合物每天两次、每天或每隔一天施用于个体。在一个优选实施例中,本发明的组合物每周两次、每周一次、每十天一次、每两周一次、每三周一次、或更优选地每四周一次、每月一次、每六周一次、每八周一次、每隔一个月一次、每三个月一次、每四个月一次、每六个月一次、每八个月一次、每九个月一次或每年一次施用于个体。还考虑了配制用于贮存施用(例如肌肉内、皮下、玻璃体内)的组合物和脂质体,以在长时间内递送或释放mRNA。优选地,所采用的延长释放手段与对mRNA进行的修饰组合,以增强稳定性。
本文还考虑了包含本文公开的一种或多种脂质体的冻干药物组合物和使用此类组合物的相关方法,如例如在2011年6月8日提交的美国临时申请第61/494,882号中所公开的,所述美国临时申请的教示内容以全文引用的方式并入本文中。例如,根据本发明的冻干药物组合物可在施用前复原或可在体内复原。例如,冻干的药物组合物可配制成适当的剂型(例如皮内剂型,如圆盘、棒或膜)并施用以使得剂型通过个体的体液在体内随时间推移而再水合。
提供的脂质体和组合物可施用于任何所需组织。在一些实施例中,通过提供的脂质体或组合物递送的mRNA在其中施用脂质体和/或组合物的组织中表达。在一些实施例中,递送的mRNA在与其中施用脂质体和/或组合物的组织不同的组织中表达。递送的mRNA可在其中递送和/或表达的示例性组织包括但不限于肺、肝、肾、心脏、脾、血清、脑、骨骼肌、淋巴结、皮肤和/或脑脊髓液。
在一些实施例中,靶组织是肺。在一些实施例中,靶组织是右肺或左肺的上(即上部)叶。在一些实施例中,靶组织是右肺或左肺的下(即下部)叶。在一些实施例中,靶组织是右肺的中叶。
在一些实施例中,靶组织是右肺的尖段或左肺的尖后段。在一些实施例中,靶组织是右肺的后段。在一些实施例中,靶组织是右肺或左肺的前段。在一些实施例中,靶组织是右肺或左肺的上段。在一些实施例中,靶组织是右肺或左肺的外侧底段。在一些实施例中,靶组织是右肺的前底段。在一些实施例中,靶组织是左肺的前内侧底段。在一些实施例中,靶组织是右肺的外侧段。在一些实施例中,靶组织是右肺的内侧段。在一些实施例中,靶组织是左肺的上舌段。在一些实施例中,靶组织是左肺的下舌段。在一些实施例中,靶组织是右肺或左肺的后底段。在一些实施例中,靶组织是右肺的内侧底段。
在特定实施例中,靶组织是肺中的上皮细胞。在一些实施例中,靶组织是肺中的平滑肌细胞。在一些实施例中,靶组织是胰管上皮细胞。在一些实施例中,靶组织是胆管上皮细胞。在一些实施例中,靶组织是唾液腺上皮细胞。在一些实施例中,靶组织是肾上皮细胞。在一些实施例中,靶组织是汗腺的汗腺分泌旋管中的β-S细胞。在一些实施例中,靶组织是生殖道上皮细胞。
根据各种实施例,可调整递送的mRNA的表达时机,以适应特定的医学需要。在一些实施例中,由递送的mRNA编码的蛋白质的表达在单次施用所提供的脂质体或组合物后在血清或靶组织中可检测1、2、3、6、12、18、24、30、36、42、48、54、60、66和/或72小时。在一些实施例中,由递送的mRNA编码的蛋白质的表达在单次施用所提供的脂质体或组合物后在血清或靶组织中可检测1天、2天、3天、4天、5天、6天和/或7天。在一些实施例中,由mRNA编码的蛋白质的表达在单次施用所提供的脂质体或组合物后在血清或靶组织中可检测1周、2周、3周和/或4周。在一些实施例中,由mRNA编码的蛋白质的表达在单次施用所提供的脂质体或组合物后的一个月或更长时间后可检测。
在一些实施例中,在递送到肺(例如通过喷雾)的如本文所提供的制剂中的mRNA(例如编码CFTR蛋白)(例如囊封在由3种不同脂质组分组成的脂质纳米颗粒中,其中一种是基于固醇的阳离子脂质)在肺组织中表达至少7天、至少14天、至少21天、或至少28天。在一些实施例中,在递送到肺(例如通过喷雾)的如本文所提供的制剂中的mRNA(例如编码CFTR蛋白)(例如囊封在由3种不同脂质组分组成的脂质纳米颗粒中,其中一种是基于固醇的阳离子脂质)在肺组织中表达至多7天、至多14天、至多21天、或至多28天。在一些实施例中,在递送到肺(例如通过喷雾)的如本文所提供的制剂中的mRNA(例如编码CFTR蛋白)(例如囊封在由3种不同脂质组分组成的脂质纳米颗粒中,其中一种是基于固醇的阳离子脂质)所编码的蛋白质(例如CFTR)在肺组织中表达至少7天、至少14天、至少21天、或至少28天。在一些实施例中,在递送到肺(例如通过喷雾)的如本文所提供的制剂中的mRNA(例如编码CFTR蛋白)(例如囊封在由3种不同脂质组分组成的脂质纳米颗粒中,其中一种是基于固醇的阳离子脂质)所编码的蛋白质(例如CFTR)在肺组织中表达至多7天、至多14天、至多21天、或至多28天。
本发明可用于在各种剂量下递送mRNA。在一些实施例中,mRNA以介于约0.1-5.0mg/kg体重,例如约0.1-4.5、0.1-4.0、0.1-3.5、0.1-3.0、0.1-2.5、0.1-2.0、0.1-1.5、0.1-1.0、0.1-0.5、0.1-0.3、0.3-5.0、0.3-4.5、0.3-4.0、0.3-3.5、0.3-3.0、0.3-2.5、0.3-2.0、0.3-1.5、0.3-1.0、0.3-0.5、0.5-5.0、0.5-4.5、0.5-4.0、0.5-3.5、0.5-3.0、0.5-2.5、0.5-2.0、0.5-1.5或0.5-1.0mg/kg体重范围内的剂量施用。在一些实施例中,mRNA以等于或小于约5.0、4.5、4.0、3.5、3.0、2.5、2.0、1.5、1.0、0.8、0.6、0.5、0.4、0.3、0.2或0.1mg/kg体重的剂量施用。
实例
虽然已根据某些实施例特异性地描述了本发明的某些化合物、组合物和方法,但以下实例仅用于说明本发明,且不打算对其进行限制。
脂质材料
本文所述的制剂包括采用阳离子脂质、辅助脂质和PEGy改性脂质的改变比率的多组分脂质混合物,其被设计成囊封各种基于核酸的材料。阳离子脂质可包括(但非排他地)基于固醇的阳离子脂质、ICE(咪唑胆固醇酯)、1,2-二油酰基-3-二甲基铵-丙烷(DODAP)、1,2-二油酰基-3-三甲基铵-丙烷(DOTAP)、1,2-二油基氧基-3-二甲基氨基丙烷(DODMA)、1,2-二-O-十八烯基-3-三甲基铵丙烷(DOTMA)、C12-200、DLinSSDMA、Target 24等。辅助脂质可包括(但非排他地)DSPC(1,2-二硬脂酰基-sn-甘油-3-磷酸胆碱)、DPPC(1,2-二棕榈酰基-sn-甘油-3-磷酸胆碱)、DOPE(1,2-二油基-sn-甘油-3-磷酸乙醇胺)、DPPE(1,2-二棕榈酰基-sn-甘油-3-磷酸乙醇胺)、DMPE(1,2-二肉豆蔻酰基-sn-甘油-3-磷酸乙醇胺)、DOPG(2-二油酰基-sn-甘油-3-磷酸-(1'-rac-甘油))、胆固醇等。PEG改性脂质可包括(但非排他地)共价连接至具有C6-C20长度的烷基链的脂质、长度高达5kDa的聚(乙)二醇链。
N/P比是阳离子型载体的氮(胺)基团的摩尔比:mRNA的磷酸酯基团的摩尔比。
信使RNA材料
本文所述的制剂包括信使RNA(mRNA)。mRNA可(但非排他地)编码人囊性纤维化跨膜传导调节因子(CFTR)和萤火虫荧光素酶(FFL)。本文公开的示例性mRNA编码序列包括SEQID NO:1、SEQ ID NO:2和SEQ ID NO:3。本文公开的示例性的5'UTR mRNA序列是SEQ ID NO:5。本文公开的示例性的3'UTR mRNA序列是SEQ ID NO:6和SEQ ID NO:7。本文公开的示例性全长mRNA序列(即编码序列加5'UTR和3'UTR)包括SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10和SEQ ID NO:11。
包含不同阳离子脂质的示例性LNP制剂
在一些实施例中,包含基于固醇的阳离子脂质、辅助脂质和PEG或PEG改性脂质的新型脂质组合物的一种特定应用是mRNA,如CFTR mRNA的肺部递送。由ICE脂质、DOPE脂质和DMG-PEG脂质(摩尔比为60:35:5)构成的脂质纳米颗粒制剂表现出针对mRNA,例如CFTRmRNA的高百分比囊封值(>80%),如通过mRNA的基于荧光的检测所测定。如表5中所示,这种特定的制剂组合物是优异的,因为具有DOPE和DMG-PEG 2K的常规pH可滴定阳离子脂质不显示此类高百分比囊封。
pH可滴定阳离子脂质 | mRNA囊封% |
ICE | 90 |
DODMA | 51 |
DODAP | 49 |
C12-200 | 39 |
DLinSSDMA | 57 |
Target 24 | 59 |
表5.基于固醇的阳离子脂质纳米颗粒制剂和其它基于阳离子脂质的纳米颗粒制剂的mRNA的mRNA百分比。所有脂质制剂都用阳离子脂质组合物DOPE:DMG-PEG 2K(摩尔比60:35:5)制备。
所有这些示例性制剂都制备为5mg mRNALNP制备物。
制剂实例#1(ICE脂质):
将ICE、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至15mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有60mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒,这是由静电引力和范德华力(van derwall force)驱动的自组装过程。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。最终浓度:0.5mg/mL CFTR mRNA(囊封)。mRNA囊封%:90%。尺寸:60nm。PDI:0.14。
制剂实例#2(DODMA脂质):
将DODMA、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至15mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有60mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒,这是由静电引力和范德华力驱动的自组装过程。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。最终浓度:0.5mg/mLCFTR mRNA(囊封)。mRNA囊封%:51%。尺寸:62nm。PDI:0.18。
制剂实例#3(DODAP脂质):
将DODAP、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至15mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有60mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒,这是由静电引力和范德华力驱动的自组装过程。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。最终浓度:0.5mg/mLCFTR mRNA(囊封)。mRNA囊封%:49%。尺寸:78nm。PDI:0.19。
制剂实例#4(C12-200脂质):
将C12-200、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至15mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有60mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒,这是由静电引力和范德华力驱动的自组装过程。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。最终浓度:0.5mg/mLCFTR mRNA(囊封)。mRNA囊封%:39%。尺寸:98nm。PDI:0.22。
制剂实例#5(DLin-SS-DMA脂质):
将DLin-SS-DMA、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至15mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有60mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒,这是由静电引力和范德华力驱动的自组装过程。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。最终浓度:0.5mg/mLCFTR mRNA(囊封)。mRNA囊封%:57%。
制剂实例#6(Target 24脂质):
将Target 24(3-(5-(双(2-羟基十二烷基)氨基)戊-2-基)-6-(5-((2-羟基十二烷基)(2-羟基十一烷基)氨基)戊-2-基)-1,4-二噁烷-2,5-二酮)、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至15mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH4.5),以具有60mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒,这是由静电引力和范德华力驱动的自组装过程。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。最终浓度:0.5mg/mLCFTR mRNA(囊封)。mRNA囊封%:59%。尺寸:92nm。PDI:0.24。
示例性ICE配制方案
A.制剂实例-1mg规模,N/P=4
将ICE、DOPE、胆固醇和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至3mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5)。将脂质溶液快速注入mRNA水溶液中且振荡,以得到20%乙醇中的最终悬浮液。将所得的纳米颗粒悬浮液过滤、渗滤、浓缩并冷冻储存,直到进一步使用。在0.5mg/mL CFTR mRNA(囊封)下测定制剂的最终浓度。制剂具有约89%mRNA囊封(89%的脂质纳米颗粒含有mRNA),并且脂质纳米颗粒具有约67nm的平均尺寸与0.19的多分散性指数(PDI)。
B.制剂实例-15mg规模,N/P=2
将ICE、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至45mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有180mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒,这是由静电引力和范德华力驱动的自组装过程。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。在0.5mg/mL CFTR mRNA(囊封)下测定制剂的最终浓度。脂质纳米颗粒具有约70nm的平均尺寸与0.17的PDI。
C.制剂实例-15mg规模,N/P=4
将ICE、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至45mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有180mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。在0.5mg/mL CFTR mRNA(囊封)下测定制剂的最终浓度。制剂具有约85%mRNA囊封,并且脂质纳米颗粒具有约65nm的平均尺寸与0.13的PDI。
D.制剂实例-30mg规模,N/P=4
将ICE、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至90mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有360mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。使用TFF系统对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。在0.5mg/mL CFTR mRNA(囊封)下测定制剂的最终浓度。制剂具有约86%mRNA囊封,并且脂质纳米颗粒具有约89nm的平均尺寸与0.12的PDI。
E.制剂实例-30mg规模,3%PEG,N/P=4
将ICE、DOPE和DMG-PEG2K(摩尔比为60:37:3)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至90mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有360mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。使用TFF系统对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。在0.5mg/mL CFTR mRNA(囊封)下测定制剂的最终浓度。脂质纳米颗粒具有约94nm的平均尺寸与0.15的PDI。
F.制剂实例-60mg规模,N/P=4
将ICE、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至180mL最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有720mL的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。使用TFF系统对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。在0.5mg/mL CFTR STOP mRNA(囊封)下测定制剂的最终浓度。制剂具有约97%mRNA囊封,并且脂质纳米颗粒具有约67nm的平均尺寸与0.12的PDI。
G.制剂实例-3.5克规模,N/P=4
将ICE、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至42L最终体积。分开地,由1mg/mL储备溶液制备CFTR mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有42L的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。使用TFF系统对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。在0.5mg/mL CFTR mRNA(囊封)下测定制剂的最终浓度。制剂具有约91%mRNA囊封,并且脂质纳米颗粒具有约50nm的平均尺寸与0.17的PDI。
H.制剂实例-200mg规模,N/P=4
将ICE、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至600mL最终体积。分开地,由1mg/mL储备溶液制备FFL mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有2.4L的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。使用TFF系统对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。在0.5mg/mL CFTR mRNA(囊封)下测定制剂的最终浓度。制剂具有约95%mRNA囊封,并且脂质纳米颗粒具有约59nm的平均尺寸与0.15的PDI。
I.制剂实例-200mg规模,N/P=4
将ICE、DOPE和DMG-PEG2K(摩尔比为60:35:5)的10mg/mL乙醇溶液的等分试样混合,并且用乙醇稀释至600mL最终体积。分开地,由1mg/mL储备溶液制备hCFTR-STOP mRNA的水性缓冲溶液(10mM柠檬酸盐/150mM NaCl,pH 4.5),以具有2.4L的最终体积。
在此过程中,将脂质溶解于乙醇中并且使用泵系统混合溶解于柠檬酸盐缓冲液中的mRNA。两种流的瞬时混合导致形成脂质纳米颗粒。所得制剂混合物在含有20%乙醇的柠檬酸盐缓冲液中。使用TFF系统对制剂进行缓冲液交换,并且将所得制剂调节至所需mRNA浓度并冷冻储存直到进一步使用。在0.5mg/mL CFTR STOP mRNA(囊封)下测定制剂的最终浓度。制剂具有约88%mRNA囊封,并且脂质纳米颗粒具有约57nm的平均尺寸与0.18的PDI。
肺部递送ICE LNP
进行若干示例性研究以展示来自囊封密码子优化的hCFTR mRNA的基于ICE的LNP的成功的mRNA递送和后续人CFTR蛋白产生。使用抗人CFTR抗体对所有肺切片进行免疫组织化学分析,以特异性检测经处理的动物肺内的人CFTR蛋白。
实例1.在气管内施用后大鼠肺中hCFTR的体内表达
图1显示在肺部递送通过上文所述的方法制备的hCFTR mRNA脂质纳米颗粒(制剂实例#1)后24小时,小鼠肺中的hCFTR蛋白的示例性免疫组织化学检测。
经由微型喷雾器(气管内气溶胶)向小鼠施用含有hCFTR mRNA的LNP制剂。使用ICE脂质作为阳离子脂质而制得LNP制剂。通过免疫组织化学染色对来自这些小鼠的固定肺组织分析hCFTR蛋白的存在。
在整个肺,包括支气管上皮细胞和肺泡区两者中检测到蛋白质。相比于未处理的对照小鼠的肺中不具有棕色染色,在所有mRNA脂质纳米颗粒测试品组中观察到阳性(棕色)染色。图A描绘了在10×放大率下的未处理的小鼠肺。图B描绘了在20×放大率下的未处理的小鼠肺。图C描绘了在10×放大率下的密码子优化的hCFTR(CO-hCFTR)mRNAICE LNP处理的小鼠肺。图D描绘了在20×放大率下的CO-hCFTR mRNA ICE LNP处理的小鼠肺。
实例2.在喷雾施用后大鼠肺中hCFTR的体内表达
图2显示在肺部递送通过上文所述的方法制备的hCFTR mRNA脂质纳米颗粒(制剂实例#1)后24小时,小鼠肺中的hCFTR蛋白的示例性免疫组织化学检测。
经由喷雾向小鼠施用含有hCFTR mRNA的LNP制剂。使用ICE脂质作为阳离子脂质而制得LNP制剂。通过免疫组织化学染色对来自这些小鼠的固定肺组织分析hCFTR蛋白的存在。
在整个肺,包括支气管上皮细胞和肺泡区两者中检测到蛋白质。相比于未处理的对照小鼠的肺中不具有棕色染色,在所有mRNA脂质纳米颗粒测试品组中观察到阳性(棕色)染色。图A描绘了在10×放大率下的未处理的小鼠肺。图B描绘了在20×放大率下的未处理的小鼠肺。图C描绘了在10×放大率下的密码子优化的hCFTR(CO-hCFTR)mRNAICE LNP处理的小鼠肺。图D描绘了在20×放大率下的CO-hCFTR mRNA ICE LNP处理的小鼠肺。
实例3.大鼠肺中hCFTR的体内表达(10mg,5%PEG)
图3显示在肺部递送使用基于ICE的脂质纳米颗粒通过上文所述的方法制备的密码子优化的hCFTR(CO-hCFTR)mRNA脂质纳米颗粒之后24小时,大鼠肺中的hCFTR蛋白的示例性免疫组织化学分析。
在一些代表性研究中,将大鼠置于允许大鼠完全活动的气溶胶室中。如下产生气溶胶:通过喷洒10mg(如根据囊封的mRNA所测量)囊封于ICE LNP(含有5%[mol]PEG改性脂质)中的CO-hCFTR mRNA,所述ICE LNP填充含有大鼠的室。大鼠在给定的时段内暴露,在此期间,通过正常呼吸将气溶胶自由吸入肺中。暴露后,将大鼠移出并放回其笼中。接着在暴露后24小时处死大鼠,收获肺部(将一部分快速冷冻并且将独立部分固定在10%NBF中,并包埋于石蜡中)进行分析。通过免疫组织化学染色对来自这些大鼠的固定肺组织分析hCFTR蛋白的存在。
在整个肺,包括支气管上皮细胞和肺泡区中检测到蛋白质,如图3中所示。在mRNA脂质纳米颗粒测试品组中观察到阳性(棕色)染色。图A描绘了低放大率下的CO-hCFTR mRNAICE LNP处理的大鼠肺。图B描绘了10×放大率下的CO-hCFTR mRNA ICE LNP处理的大鼠肺。图C描绘了20×放大率下的CO-hCFTR mRNA ICE LNP处理的大鼠肺。
实例4.大鼠肺中hCFTR的体内表达(50pg mRNA,5%PEG)
图4显示在肺部递送使用基于ICE的脂质纳米颗粒通过上文所述的方法制备的CO-hCFTR mRNA脂质纳米颗粒之后24小时,大鼠肺中的hCFTR蛋白的示例性免疫组织化学分析。
在一些代表性研究中,经由微型喷雾器设备使用直接滴注使大鼠暴露于CO-hCFTRmRNA ICE LNP。在向大鼠气管内施用50pg CO-hCFTR mRNA囊封的ICE LNP(含有5%[mol]PEG改性脂质)期间经由微型喷雾器设备产生气溶胶。接着在暴露后24小时处死大鼠,收获肺部(将一部分快速冷冻并且将独立部分固定在10%NBF中,并包埋于石蜡中)进行分析。通过免疫组织化学染色对来自这些大鼠的固定肺组织分析hCFTR蛋白的存在。
在整个肺,包括支气管上皮细胞和肺泡区中检测到蛋白质,如图4中所示。在mRNA脂质纳米颗粒测试品组中观察到阳性(棕色)染色。图A描绘了低放大率下的CO-hCFTR mRNAICE LNP处理的大鼠肺。图B描绘了10×放大率下的CO-hCFTR mRNA ICE LNP处理的大鼠肺。图C描绘了20×放大率下的CO-hCFTR mRNA ICE LNP处理的大鼠肺。
实例5.大鼠肺中hCFTR的体内表达(10mg,3%PEG)
图5显示在肺部递送使用基于ICE的脂质纳米颗粒通过上文所述的方法制备的CO-hCFTR mRNA脂质纳米颗粒之后24小时,大鼠肺中的hCFTR蛋白的示例性免疫组织化学分析。
在一些代表性研究中,将大鼠置于允许大鼠完全活动的气溶胶室中。如下产生气溶胶:通过喷洒10mg囊封于ICE LNP(含有3%[mol]PEG改性脂质)中的CO-hCFTR mRNA,所述ICE LNP填充含有大鼠的室。大鼠在给定的时段内暴露,在此期间,通过正常呼吸将气溶胶自由吸入肺中。暴露后,将大鼠移出并放回其笼中。接着在暴露后24小时处死大鼠,收获肺部(将一部分快速冷冻并且将独立部分固定在10%NBF中,并包埋于石蜡中)进行分析。通过免疫组织化学染色对来自这些大鼠的固定肺组织分析hCFTR蛋白的存在。
在整个肺,包括支气管上皮细胞和肺泡区中检测到蛋白质,如图5中所示。在mRNA脂质纳米颗粒测试品组中观察到阳性(棕色)染色。图A描绘了低放大率下的CO-hCFTR mRNAICE LNP处理的大鼠肺。图B描绘了10×放大率下的CO-hCFTR mRNA ICE LNP处理的大鼠肺。图C描绘了20×放大率下的CO-hCFTR mRNA ICE LNP处理的大鼠肺。
实例6.大鼠肺中hCFTR的体内表达(50pg mRNA,3%PEG)
图6显示在肺部递送使用基于ICE的脂质纳米颗粒通过上文所述的方法制备的CO-hCFTR mRNA脂质纳米颗粒之后24小时,大鼠肺中的hCFTR蛋白的示例性免疫组织化学分析。
在一些代表性研究中,经由微型喷雾器设备使用直接滴注使大鼠暴露于CO-hCFTRmRNA ICE LNP。在向大鼠气管内施用50pg CO-hCFTR mRNA囊封的ICE LNP(含有3%[mol]PEG改性脂质)期间经由微型喷雾器设备产生气溶胶。接着在暴露后24小时处死大鼠,收获肺部(将一部分快速冷冻并且将独立部分固定在10%NBF中,并包埋于石蜡中)进行分析。通过免疫组织化学染色对来自这些大鼠的固定肺组织分析hCFTR蛋白的存在。
在整个肺,包括支气管上皮细胞和肺泡区中检测到蛋白质,如图6中所示。在mRNA脂质纳米颗粒测试品组中观察到阳性(棕色)染色。图A描绘了低放大率下的CO-hCFTR mRNAICE LNP处理的大鼠肺。图B描绘了10×放大率下的CO-hCFTR mRNA ICE LNP处理的大鼠肺。图C描绘了20×放大率下的CO-hCFTR mRNA ICE LNP处理的大鼠肺。
实例7.小鼠肺中hCFTR的体内表达(10mg,N/P=2)
图7显示在肺部递送使用基于ICE的脂质纳米颗粒通过上文所述的方法制备的CO-hCFTR mRNA脂质纳米颗粒之后24小时,CFTR KO小鼠肺中的hCFTR蛋白的示例性免疫组织化学分析。
在一些代表性研究中,将小鼠置于允许小鼠完全活动的气溶胶室中。如下产生气溶胶:通过喷洒10mg囊封于ICE LNP(N/P=2)中的CO-hCFTR mRNA,所述ICE LNP填充含有小鼠的室。小鼠在给定的时段内暴露,在此期间,通过正常呼吸将气溶胶自由吸入肺中。暴露后,将小鼠移出并放回其笼中。接着在暴露后24小时处死小鼠,收获肺部(将一部分快速冷冻并且将独立部分固定在10%NBF中,并包埋于石蜡中)进行分析。通过免疫组织化学染色对来自这些小鼠的固定肺组织分析hCFTR蛋白的存在。
在整个肺,包括支气管上皮细胞和肺泡区中检测到蛋白质,如图7中所示。在mRNA脂质纳米颗粒测试品组中观察到阳性(棕色)染色。图A描绘了低放大率下的CO-hCFTR mRNAICE LNP处理的小鼠肺。图B描绘了10×放大率下的CO-hCFTR mRNA ICE LNP处理的小鼠肺。图C描绘了20×放大率下的CO-hCFTR mRNA ICE LNP处理的小鼠肺。
实例8.小鼠肺中hCFTR的体内表达(10mg,N/P=4)
图8显示在肺部递送使用基于ICE的脂质纳米颗粒通过上文所述的方法制备的CO-hCFTR mRNA脂质纳米颗粒之后24小时,CFTR KO小鼠肺中的hCFTR蛋白的示例性免疫组织化学分析。
在一些代表性研究中,将小鼠置于允许小鼠完全活动的气溶胶室中。如下产生气溶胶:通过喷洒10mg囊封于ICE LNP(N/P=4)中的CO-hCFTR mRNA,所述ICE LNP填充含有小鼠的室。小鼠在给定的时段内暴露,在此期间,通过正常呼吸将气溶胶自由吸入肺中。暴露后,将小鼠移出并放回其笼中。接着在暴露后24小时处死小鼠,收获肺部(将一部分快速冷冻并且将独立部分固定在10%NBF中,并包埋于石蜡中)进行分析。通过免疫组织化学染色对来自这些小鼠的固定肺组织分析hCFTR蛋白的存在。
在整个肺,包括支气管上皮细胞和肺泡区中检测到蛋白质,如图8中所示。在mRNA脂质纳米颗粒测试品组中观察到阳性(棕色)染色。图A描绘了低放大率下的CO-hCFTRmRNAICE LNP处理的小鼠肺。图B描绘了10×放大率下的CO-hCFTR mRNAICE LNP处理的小鼠肺。图C描绘了20×放大率下的CO-hCFTR mRNA ICE LNP处理的小鼠肺。在实例5(N/P=2)和实例6(N/P=4)中均观察到hCFTR蛋白的广泛分布。
实例9.具有不同制剂的小鼠肺中hCFTR的体内表达
图9显示在肺部递送使用基于ICE的脂质纳米颗粒通过上文所述的方法制备的CO-hCFTR mRNA脂质纳米颗粒之后24小时,野生型小鼠肺中的hCFTR蛋白的示例性免疫组织化学分析。
在一些代表性研究中,将几组野生型小鼠分离并置于个别气溶胶室中。每组用不同的制剂处理。制剂由囊封以下mRNA构筑体的基于ICE的LNP(N/P=4)组成:1)CO-hCFTRmRNA ICE LNP;2)CO-hCFTR“STOP”mRNA(不能翻译为蛋白质的无义突变CO-hCFTR mRNA)ICELNP;3)FFL(萤火虫荧光素酶)mRNAICE LNP;和4)缓冲液(媒剂)。
在一些实施例中,将小鼠置于允许小鼠完全活动的气溶胶室中。如下产生气溶胶:通过囊封于ICE LNP(N/P=4)中的CO-hCFTR mRNA的喷雾器,所述ICE LNP填充含有小鼠的室。使小鼠在给定的时段内暴露于总共50mg(显示于图7中)气溶胶化CO-hCFTR mRNA(如根据囊封的mRNA所测量),在此期间,通过正常呼吸将气溶胶自由吸入肺中。暴露后,将小鼠移出并放回其笼中。接着在暴露后24小时处死小鼠,收获肺部(将一部分快速冷冻并且将独立部分固定在10%NBF中,并包埋于石蜡中)进行分析。通过免疫组织化学染色对来自这些小鼠的固定肺组织分析hCFTR蛋白的存在。
在图9中,仅在CO-hCFTR mRNA LNP测试品组中观察到阳性(棕色)染色。第一图描绘了CO-hCFTR mRNAICE LNP处理的小鼠肺,其显示在整个肺,包括支气管上皮细胞和肺泡区两者中观察到的人CFTR蛋白的广泛分布。第二面板描绘了未观察到阳性(棕色)染色的CO-hCFTR“STOP”mRNAICE LNP处理的小鼠肺。第三图描绘了未观察到阳性(棕色)染色的FFLmRNA ICE LNP处理的小鼠肺。第四图描绘了未观察到阳性(棕色)染色的缓冲液处理的小鼠肺。
实例10.在不同时间点的小鼠肺中hCFTR的体内表达
图10显示在肺部递送使用基于ICE的脂质纳米颗粒通过上文所述的方法制备的CO-hCFTR mRNA脂质纳米颗粒之后的预定时间点,野生型小鼠肺中的hCFTR蛋白的示例性免疫组织化学分析。
在一些代表性研究中,在CFTR KO小鼠中进行CO-hCFTR mRNA囊封的ICE LNP处理后的hCFTR蛋白的药代动力学行为。在一些实施例中,将小鼠置于允许小鼠完全活动的气溶胶室中。如下产生气溶胶:通过喷洒囊封于ICE LNP(N/P=4)中的CO-hCFTR mRNA,所述ICELNP填充含有小鼠的室。使小鼠在给定的时段内暴露于10mg气溶胶化CO-hCFTR mRNA,在此期间,通过正常呼吸将气溶胶自由吸入肺中。暴露后,将小鼠移出并放回其笼中。在气溶胶暴露之后的预定的时间点处死经处理小鼠的所选群体。施用后处死小鼠的时间范围在30分钟至一周范围内,确切地说,30分钟、2小时、4小时、6小时、24小时、48小时、72小时和1周,如图10中所示。收获小鼠肺(将一部分快速冷冻并且将独立部分固定在10%NBF中,并包埋于石蜡中)进行分析。通过免疫组织化学染色对来自这些小鼠的固定肺组织分析hCFTR蛋白的存在。
如图10中所示,在施用后超过30分钟处死的hCFTR mRNA处理的小鼠组中,在整个肺,包括支气管上皮细胞和肺泡区两者中检测到蛋白质或阳性(棕色)染色。在hCFTR mRNA递送之后24小时、48小时和72小时观察到最阳性或棕色的染色。在生理盐水处理的对照组中未观察到阳性(棕色)染色。
实例11.在不同剂量水平下大鼠肺中hCFTR的体内表达
在一些代表性研究中,将大鼠置于允许大鼠完全活动的气溶胶室中。如下产生气溶胶:通过喷洒囊封于ICE LNP中的CO-hCFTR mRNA,所述ICE LNP填充含有大鼠的室。大鼠在给定的时段内暴露,在此期间,通过正常呼吸将气溶胶自由吸入肺中。几个组以以下的不同剂量水平给药:1)缓冲液;2)空ICE LNP(约8.0mg/kg等效物);3)ICE LNP(0.5mg/kg);4)ICE LNP(2.1mg/kg);5)ICE LNP(4.1mg/kg);和6)ICE LNP(6.2mg/kg)。
施用后,将大鼠移出并放回其笼中。此外,将每组大鼠分成在施用后的不同预定时间点处死的所选群体。施用后处死大鼠的时间范围为24小时至28天,确切地说,24小时、7天、14天和28天。
对每组中的所有经处理大鼠的快速冷冻肺进行定量PCR,其测量CO-hCFTR mRNA的拷贝/总RNA。结果表示于图11中,显示了CO-hCFTR mRNA的拷贝/微克总RNA的比率。跨施用的不同剂量和在每个剂量水平的所选处死时间点分析结果。CO-hCFTR mRNA的背景水平由对照组的(缓冲液和空LNP)值指示。随着剂量水平增加,大鼠冷冻肺切片中的CO-hCFTRmRNA的拷贝/总RNA增加。在相同剂量水平下,随着施用后的处死时间增加,大鼠冷冻肺切片中的CO-hCFTR mRNA的拷贝/总RNA减小。
图12显示外源性CO-hCFTR mRNA的拷贝/内源性CFTR mRNA的拷贝水平的比较。评估大鼠冷冻肺切片中CO-hCFTR mRNA的拷贝/内源性CFTR mRNA的拷贝的倍数增加或比率,并且作为剂量和处死时间的函数标绘。还使用定量PCR获得数据。CO-hCFTR mRNA的拷贝相比于CFTR mRNA的内源性水平的倍数增加随着剂量水平增加而增加。在相同mRNA剂量下,CO-hCFTR mRNA的拷贝相比于CFTR mRNA的内源性水平的倍数增加随着施用后的处死时间增加而减小。
还进行了大鼠肺的免疫组织化学分析,并且显示于图13中。通过免疫组织化学染色对来自这些大鼠的固定肺组织分析hCFTR蛋白的存在。观察到hCFTR蛋白的剂量依赖性增加,如通过在整个肺,包括支气管上皮细胞和肺泡区两者中的阳性(棕色)染色所确定。观察到hCFTR蛋白的减少,如通过单次施用剂量之后的时间增加超过24小时的阳性(棕色)染色所确定。然而,在hCFTR mRNAICE LNP单次暴露后28天观察到可检测水平的hCFTR蛋白。在生理盐水处理的对照组或空LNP组中未观察到阳性(棕色)染色。
实例12.hCFTR制剂对呼吸的体内作用
在一些代表性研究中,大鼠经由吸入暴露于单剂量的hCFTR mRNA。hCFTR mRNA用如上所述的ICE或用支链PEI(bPEI)配制。如图14A中所示,相对于所研究大鼠中的生理盐水对照,用bPEI配制的各种剂量的hCFTR mRNA引起不良呼吸速率增加。当处死大鼠时,还发现肺的重量也增加。相比之下,如图14B中所示,相对于所研究大鼠中的缓冲液对照,用ICE配制的各种剂量的hCFTR mRNA未引起呼吸速率增加。当处死大鼠时也没有观察到大鼠肺重量的增加。
实例13.在不同剂量水平下hCFTR的体外表达和活性
在一些代表性实验中,根据标准程序使用脂染胺将hCFTR mRNA转染到培养的费舍尔大鼠(Fischer rat)甲状腺细胞中。如图15中所示,存在由hCFTR mRNA诱导的氯离子通道活性的剂量反应。这指示hCFTR mRNA在转染细胞中产生活性CFTR蛋白。
等效物
所属领域的技术人员将认识到,或者使用不超出常规的实验就能够确定本文所述的发明的具体实施例的许多等效物。本发明的范围不打算限于以上说明书,而是如以下权利要求书所阐述:
序列表
<110> 川斯勒佰尔公司
<120> 用于递送MRNA的改进的基于ICE的脂质纳米颗粒制剂
<130> MRT-1247WO
<150> 62/420,421
<151> 2016-11-10
<150> 62/420,428
<151> 2016-11-10
<150> 62/421,021
<151> 2016-11-11
<150> 62/421,007
<151> 2016-11-11
<150> 62/464,327
<151> 2017-02-27
<150> 62/464,330
<151> 2017-02-27
<160> 11
<170> PatentIn version 3.5
<210> 1
<211> 4443
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 1
augcaacgcu cuccucuuga aaaggccucg guggugucca agcucuucuu cucguggacu 60
agacccaucc ugagaaaggg guacagacag cgcuuggagc uguccgauau cuaucaaauc 120
ccuuccgugg acuccgcgga caaccugucc gagaagcucg agagagaaug ggacagagaa 180
cucgccucaa agaagaaccc gaagcugauu aaugcgcuua ggcggugcuu uuucuggcgg 240
uucauguucu acggcaucuu ccucuaccug ggagagguca ccaaggccgu gcagccccug 300
uugcugggac ggauuauugc cuccuacgac cccgacaaca aggaagaaag aagcaucgcu 360
aucuacuugg gcaucggucu gugccugcuu uucaucgucc ggacccucuu guugcauccu 420
gcuauuuucg gccugcauca cauuggcaug cagaugagaa uugccauguu uucccugauc 480
uacaagaaaa cucugaagcu cucgagccgc gugcuugaca agauuuccau cggccagcuc 540
gugucccugc ucuccaacaa ucugaacaag uucgacgagg gccucgcccu ggcccacuuc 600
guguggaucg ccccucugca aguggcgcuu cugaugggcc ugaucuggga gcugcugcaa 660
gccucggcau ucugugggcu uggauuccug aucgugcugg cacuguucca ggccggacug 720
gggcggauga ugaugaagua cagggaccag agagccggaa agauuuccga acggcuggug 780
aucacuucgg aaaugaucga aaacauccag ucagugaagg ccuacugcug ggaagaggcc 840
auggaaaaga ugauugaaaa ccuccggcaa accgagcuga agcugacccg caaggccgcu 900
uacgugcgcu auuucaacuc guccgcuuuc uucuucuccg gguucuucgu gguguuucuc 960
uccgugcucc ccuacgcccu gauuaaggga aucauccuca ggaagaucuu caccaccauu 1020
uccuucugua ucgugcuccg cauggccgug acccggcagu ucccaugggc cgugcagacu 1080
ugguacgacu cccugggagc cauuaacaag auccaggacu uccuucaaaa gcaggaguac 1140
aagacccucg aguacaaccu gacuacuacc gaggucguga uggaaaacgu caccgccuuu 1200
ugggaggagg gauuuggcga acuguucgag aaggccaagc agaacaacaa caaccgcaag 1260
accucgaacg gugacgacuc ccucuucuuu ucaaacuuca gccugcucgg gacgcccgug 1320
cugaaggaca uuaacuucaa gaucgaaaga ggacagcucc uggcgguggc cggaucgacc 1380
ggagccggaa agacuucccu gcugauggug aucaugggag agcuugaacc uagcgaggga 1440
aagaucaagc acuccggccg caucagcuuc uguagccagu uuuccuggau caugcccgga 1500
accauuaagg aaaacaucau cuucggcgug uccuacgaug aauaccgcua ccgguccgug 1560
aucaaagccu gccagcugga agaggauauu ucaaaguucg cggagaaaga uaacaucgug 1620
cugggcgaag gggguauuac cuugucgggg ggccagcggg cuagaaucuc gcuggccaga 1680
gccguguaua aggacgccga ccuguaucuc cuggacuccc ccuucggaua ccuggacguc 1740
cugaccgaaa aggagaucuu cgaaucgugc gugugcaagc ugauggcuaa caagacucgc 1800
auccucguga ccuccaaaau ggagcaccug aagaaggcag acaagauucu gauucugcau 1860
gagggguccu ccuacuuuua cggcaccuuc ucggaguugc agaacuugca gcccgacuuc 1920
ucaucgaagc ugauggguug cgacagcuuc gaccaguucu ccgccgaaag aaggaacucg 1980
auccugacgg aaaccuugca ccgcuucucu uuggaaggcg acgccccugu gucauggacc 2040
gagacuaaga agcagagcuu caagcagacc ggggaauucg gcgaaaagag gaagaacagc 2100
aucuugaacc ccauuaacuc cauccgcaag uucucaaucg ugcaaaagac gccacugcag 2160
augaacggca uugaggagga cuccgacgaa ccccuugaga ggcgccuguc ccuggugccg 2220
gacagcgagc agggagaagc cauccugccu cggauuuccg ugaucuccac ugguccgacg 2280
cuccaagccc ggcggcggca guccgugcug aaccugauga cccacagcgu gaaccagggc 2340
caaaacauuc accgcaagac uaccgcaucc acccggaaag ugucccuggc accucaagcg 2400
aaucuuaccg agcucgacau cuacucccgg agacugucgc aggaaaccgg gcucgaaauu 2460
uccgaagaaa ucaacgagga ggaucugaaa gagugcuucu ucgacgauau ggagucgaua 2520
cccgccguga cgacuuggaa cacuuaucug cgguacauca cugugcacaa gucauugauc 2580
uucgugcuga uuuggugccu ggugauuuuc cuggccgagg ucgcggccuc acugguggug 2640
cucuggcugu ugggaaacac gccucugcaa gacaagggaa acuccacgca cucgagaaac 2700
aacagcuaug ccgugauuau cacuuccacc uccucuuauu acguguucua caucuacguc 2760
ggaguggcgg auacccugcu cgcgaugggu uucuucagag gacugccgcu gguccacacc 2820
uugaucaccg ucagcaagau ucuucaccac aagauguugc auagcgugcu gcaggccccc 2880
auguccaccc ucaacacucu gaaggccgga ggcauucuga acagauucuc caaggacauc 2940
gcuauccugg acgaucuccu gccgcuuacc aucuuugacu ucauccagcu gcugcugauc 3000
gugauuggag caaucgcagu gguggcggug cugcagccuu acauuuucgu ggccacugug 3060
ccggucauug uggcguucau caugcugcgg gccuacuucc uccaaaccag ccagcagcug 3120
aagcaacugg aauccgaggg acgauccccc aucuucacuc accuugugac gucguugaag 3180
ggacugugga cccuccgggc uuucggacgg cagcccuacu ucgaaacccu cuuccacaag 3240
gcccugaacc uccacaccgc caauugguuc cuguaccugu ccacccugcg gugguuccag 3300
augcgcaucg agaugauuuu cgucaucuuc uucaucgcgg ucacauucau cagcauccug 3360
acuaccggag agggagaggg acgggucgga auaauccuga cccucgccau gaacauuaug 3420
agcacccugc agugggcagu gaacagcucg aucgacgugg acagccugau gcgaagcguc 3480
agccgcgugu ucaaguucau cgacaugccu acugagggaa aacccacuaa guccacuaag 3540
cccuacaaaa auggccagcu gagcaagguc augaucaucg aaaacuccca cgugaagaag 3600
gacgauauuu ggcccuccgg aggucaaaug accgugaagg accugaccgc aaaguacacc 3660
gagggaggaa acgccauucu cgaaaacauc agcuucucca uuucgccggg acagcggguc 3720
ggccuucucg ggcggaccgg uuccgggaag ucaacucugc ugucggcuuu ccuccggcug 3780
cugaauaccg agggggaaau ccaaauugac ggcgugucuu gggauuccau uacucugcag 3840
caguggcgga aggccuucgg cgugaucccc cagaaggugu ucaucuucuc ggguaccuuc 3900
cggaagaacc uggauccuua cgagcagugg agcgaccaag aaaucuggaa ggucgccgac 3960
gaggucggcc ugcgcuccgu gauugaacaa uuuccuggaa agcuggacuu cgugcucguc 4020
gacgggggau guguccuguc gcacggacau aagcagcuca ugugccucgc acgguccgug 4080
cucuccaagg ccaagauucu gcugcuggac gaaccuucgg cccaccugga uccggucacc 4140
uaccagauca ucaggaggac ccugaagcag gccuuugccg auugcaccgu gauucucugc 4200
gagcaccgca ucgaggccau gcuggagugc cagcaguucc uggucaucga ggagaacaag 4260
guccgccaau acgacuccau ucaaaagcuc cucaacgagc ggucgcuguu cagacaagcu 4320
auuucaccgu ccgauagagu gaagcucuuc ccgcaucgga acagcucaaa gugcaaaucg 4380
aagccgcaga ucgcagccuu gaaggaagag acugaggaag aggugcagga cacccggcuu 4440
uaa 4443
<210> 2
<211> 4443
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 2
augcagcggu ccccgcucga aaaggccagu gucgugucca aacucuucuu cucauggacu 60
cggccuaucc uuagaaaggg guaucggcag aggcuugagu ugucugacau cuaccagauc 120
cccucgguag auucggcgga uaaccucucg gagaagcucg aacgggaaug ggaccgcgaa 180
cucgcgucua agaaaaaccc gaagcucauc aacgcacuga gaaggugcuu cuucuggcgg 240
uucauguucu acgguaucuu cuuguaucuc ggggagguca caaaagcagu ccaaccccug 300
uuguuggguc gcauuaucgc cucguacgac cccgauaaca aagaagaacg gagcaucgcg 360
aucuaccucg ggaucggacu guguuugcuu uucaucguca gaacacuuuu guugcaucca 420
gcaaucuucg gccuccauca caucgguaug cagaugcgaa ucgcuauguu uagcuugauc 480
uacaaaaaga cacugaaacu cucgucgcgg guguuggaua agauuuccau cggucaguug 540
gugucccugc uuaguaauaa ccucaacaaa uucgaugagg gacuggcgcu ggcacauuuc 600
guguggauug ccccguugca agucgcccuu uugaugggcc uuauuuggga gcuguugcag 660
gcaucugccu uuuguggccu gggauuucug auuguguugg cauuguuuca ggcugggcuu 720
gggcggauga ugaugaagua ucgcgaccag agagcgggua aaaucucgga aagacucguc 780
aucacuucgg aaaugaucga aaacauccag ucggucaaag ccuauugcug ggaagaagcu 840
auggagaaga ugauugaaaa ccuccgccaa acugagcuga aacugacccg caaggcggcg 900
uauguccggu auuucaauuc gucagcguuc uucuuuuccg gguucuucgu ugucuuucuc 960
ucgguuuugc cuuaugccuu gauuaagggg auuauccucc gcaagauuuu caccacgauu 1020
ucguucugca uuguauugcg cauggcagug acacggcaau uuccgugggc cgugcagaca 1080
ugguaugacu cgcuuggagc gaucaacaaa auccaagacu ucuugcaaaa gcaagaguac 1140
aagacccugg aguacaaucu uacuacuacg gagguaguaa uggagaaugu gacggcuuuu 1200
ugggaagagg guuuuggaga acuguuugag aaagcaaagc agaauaacaa caaccgcaag 1260
accucaaaug gggacgauuc ccuguuuuuc ucgaacuucu cccugcucgg aacacccgug 1320
uugaaggaca ucaauuucaa gauugagagg ggacagcuuc ucgcgguagc gggaagcacu 1380
ggugcgggaa aaacuagccu cuugauggug auuauggggg agcuugagcc cagcgagggg 1440
aagauuaaac acuccgggcg uaucucauuc uguagccagu uuucauggau caugcccgga 1500
accauuaaag agaacaucau uuucggagua uccuaugaug aguaccgaua cagaucgguc 1560
auuaaggcgu gccaguugga agaggacauu ucuaaguucg ccgagaagga uaacaucguc 1620
uugggagaag gggguauuac auugucggga gggcagcgag cgcggaucag ccucgcgaga 1680
gcgguauaca aagaugcaga uuuguaucug cuugauucac cguuuggaua ccucgacgua 1740
uugacagaaa aagaaaucuu cgagucgugc guguguaaac uuauggcuaa uaagacgaga 1800
auccugguga caucaaaaau ggaacaccuu aagaaggcgg acaagauccu gauccuccac 1860
gaaggaucgu ccuacuuuua cggcacuuuc ucagaguugc aaaacuugca gccggacuuc 1920
ucaagcaaac ucauggggug ugacucauuc gaccaguuca gcgcggaacg gcggaacucg 1980
aucuugacgg aaacgcugca ccgauucucg cuugagggug augccccggu aucguggacc 2040
gagacaaaga agcagucguu uaagcagaca ggagaauuug gugagaaaag aaagaacagu 2100
aucuugaauc cuauuaacuc aauucgcaag uucucaaucg uccagaaaac uccacugcag 2160
augaauggaa uugaagagga uucggacgaa ccccuggagc gcaggcuuag ccucgugccg 2220
gauucagagc aaggggaggc cauucuuccc cggauuucgg ugauuucaac cggaccuaca 2280
cuucaggcga ggcgaaggca auccgugcuc aaccucauga cgcauucggu aaaccagggg 2340
caaaacauuc accgcaaaac gacggccuca acgagaaaag ugucacuugc accccaggcg 2400
aauuugacug aacucgacau cuacagccgu aggcuuucgc aagaaaccgg acuugagauc 2460
agcgaagaaa ucaaugaaga agauuugaaa gaguguuucu uugaugacau ggaaucaauc 2520
ccagcgguga caacguggaa cacauacuug cguuacauca cggugcacaa guccuugauu 2580
uucguccuca ucuggugucu cgugaucuuu cucgcugagg ucgcagcguc acuugugguc 2640
cucuggcugc uugguaauac gcccuugcaa gacaaaggca auucuacaca cucaagaaac 2700
aauuccuaug ccgugauuau cacuucuaca agcucguauu acguguuuua caucuacgua 2760
ggaguggccg acacucugcu cgcgaugggu uucuuccgag gacucccacu cguucacacg 2820
cuuaucacug ucuccaagau ucuccaccau aagaugcuuc auagcguacu gcaggcuccc 2880
auguccaccu ugaauacgcu caaggcggga gguauuuuga aucgcuucuc aaaagauauu 2940
gcaauuuugg augaccuucu gccccugacg aucuucgacu ucauccaguu guugcugauc 3000
gugauugggg cuauugcagu agucgcuguc cuccagccuu acauuuuugu cgcgaccguu 3060
ccggugaucg uggcguuuau caugcugcgg gccuauuucu ugcagacguc acagcagcuu 3120
aagcaacugg agucugaagg gaggucgccu aucuuuacgc aucuugugac caguuugaag 3180
ggauugugga cguugcgcgc cuuuggcagg cagcccuacu uugaaacacu guuccacaaa 3240
gcgcugaauc uccauacggc aaauugguuu uuguauuuga guacccuccg augguuucag 3300
augcgcauug agaugauuuu ugugaucuuc uuuaucgcgg ugacuuuuau cuccaucuug 3360
accacgggag agggcgaggg acgggucggu auuauccuga cacucgccau gaacauuaug 3420
agcacuuugc agugggcagu gaacagcucg auugaugugg auagccugau gagguccguu 3480
ucgagggucu uuaaguucau cgacaugccg acggagggaa agcccacaaa aaguacgaaa 3540
cccuauaaga augggcaauu gaguaaggua augaucaucg agaacaguca cgugaagaag 3600
gaugacaucu ggccuagcgg gggucagaug accgugaagg accugacggc aaaauacacc 3660
gagggaggga acgcaauccu ugaaaacauc ucguucagca uuagccccgg ucagcgugug 3720
ggguugcucg ggaggaccgg gucaggaaaa ucgacguugc ugucggccuu cuugagacuu 3780
cugaauacag agggugagau ccagaucgac ggcguuucgu gggauagcau caccuugcag 3840
caguggcgga aagcguuugg aguaaucccc caaaaggucu uuaucuuuag cggaaccuuc 3900
cgaaagaauc ucgauccuua ugaacagugg ucagaucaag agauuuggaa agucgcggac 3960
gagguuggcc uucggagugu aaucgagcag uuuccgggaa aacucgacuu uguccuugua 4020
gaugggggau gcguccuguc gcaugggcac aagcagcuca ugugccuggc gcgauccguc 4080
cucucuaaag cgaaaauucu ucucuuggau gaaccuucgg cccaucugga cccgguaacg 4140
uaucagauca ucagaaggac acuuaagcag gcguuugccg acugcacggu gauucucugu 4200
gagcaucgua ucgaggccau gcucgaaugc cagcaauuuc uugucaucga agagaauaag 4260
guccgccagu acgacuccau ccagaagcug cuuaaugaga gaucauuguu ccggcaggcg 4320
auuucaccau ccgauagggu gaaacuuuuu ccacacagaa auucgucgaa gugcaagucc 4380
aaaccgcaga ucgcggccuu gaaagaagag acugaagaag aaguucaaga cacgcgucuu 4440
uaa 4443
<210> 3
<211> 4443
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 3
augcagcggu ccuagcucga aaaggccagu gucgugucca aacucuucuu cucauggacu 60
cggccuaucc uuagaaaggg guaucggcag aggcuugagu ugucugacau cuaccaguga 120
cccucgguag auucggcgga uaaccucucg gagaagcucg aacgggaaug ggaccgcgaa 180
cucgcgucua agaaaaaccc gaagcucauc aacgcacuga gaaggugcuu cuucuggcgg 240
uucauguucu acgguaucuu cuuguaucuc ggggagguca caaaagcagu ccaaccccug 300
uuguuggguc gcauuaucgc cucguacgac cccgauaaca aagaagaacg gagcaucgcg 360
aucuaccucg ggaucggacu guguuugcuu uucaucguca gaacacuuuu guugcaucca 420
gcaaucuucg gccuccauca caucgguaug cagaugcgaa ucgcuauguu uagcuugauc 480
uacaaaaaga cacugaaacu cucgucgcgg guguuggaua agauuuccau cggucaguug 540
gugucccugc uuaguaauaa ccucaacaaa uucgaugagg gacuggcgcu ggcacauuuc 600
guguggauug ccccguugca agucgcccuu uugaugggcc uuauuuggga gcuguugcag 660
gcaucugccu uuuguggccu gggauuucug auuguguugg cauuguuuca ggcugggcuu 720
gggcggauga ugaugaagua ucgcgaccag agagcgggua aaaucucgga aagacucguc 780
aucacuucgg aaaugaucga aaacauccag ucggucaaag ccuauugcug ggaagaagcu 840
auggagaaga ugauugaaaa ccuccgccaa acugagcuga aacugacccg caaggcggcg 900
uauguccggu auuucaauuc gucagcguuc uucuuuuccg gguucuucgu ugucuuucuc 960
ucgguuuugc cuuaugccuu gauuaagggg auuauccucc gcaagauuuu caccacgauu 1020
ucguucugca uuguauugcg cauggcagug acacggcaau uuccgugggc cgugcagaca 1080
ugguaugacu cgcuuggagc gaucaacaaa auccaagacu ucuugcaaaa gcaagaguac 1140
aagacccugg aguacaaucu uacuacuacg gagguaguaa uggagaaugu gacggcuuuu 1200
ugggaagagg guuuuggaga acuguuugag aaagcaaagc agaauaacaa caaccgcaag 1260
accucaaaug gggacgauuc ccuguuuuuc ucgaacuucu cccugcucgg aacacccgug 1320
uugaaggaca ucaauuucaa gauugagagg ggacagcuuc ucgcgguagc gggaagcacu 1380
ggugcgggaa aaacuagccu cuugauggug auuauggggg agcuugagcc cagcgagggg 1440
aagauuaaac acuccgggcg uaucucauuc uguagccagu uuucauggau caugcccgga 1500
accauuaaag agaacaucau uuucggagua uccuaugaug aguaccgaua cagaucgguc 1560
auuaaggcgu gccaguugga agaggacauu ucuaaguucg ccgagaagga uaacaucguc 1620
uugggagaag gggguauuac auugucggga gggcagcgag cgcggaucag ccucgcgaga 1680
gcgguauaca aagaugcaga uuuguaucug cuugauucac cguuuggaua ccucgacgua 1740
uugacagaaa aagaaaucuu cgagucgugc guguguaaac uuauggcuaa uaagacgaga 1800
auccugguga caucaaaaau ggaacaccuu aagaaggcgg acaagauccu gauccuccac 1860
gaaggaucgu ccuacuuuua cggcacuuuc ucagaguugc aaaacuugca gccggacuuc 1920
ucaagcaaac ucauggggug ugacucauuc gaccaguuca gcgcggaacg gcggaacucg 1980
aucuugacgg aaacgcugca ccgauucucg cuugagggug augccccggu aucguggacc 2040
gagacaaaga agcagucguu uaagcagaca ggagaauuug gugagaaaag aaagaacagu 2100
aucuugaauc cuauuaacuc aauucgcaag uucucaaucg uccagaaaac uccacugcag 2160
augaauggaa uugaagagga uucggacgaa ccccuggagc gcaggcuuag ccucgugccg 2220
gauucagagc aaggggaggc cauucuuccc cggauuucgg ugauuucaac cggaccuaca 2280
cuucaggcga ggcgaaggca auccgugcuc aaccucauga cgcauucggu aaaccagggg 2340
caaaacauuc accgcaaaac gacggccuca acgagaaaag ugucacuugc accccaggcg 2400
aauuugacug aacucgacau cuacagccgu aggcuuucgc aagaaaccgg acuugagauc 2460
agcgaagaaa ucaaugaaga agauuugaaa gaguguuucu uugaugacau ggaaucaauc 2520
ccagcgguga caacguggaa cacauacuug cguuacauca cggugcacaa guccuugauu 2580
uucguccuca ucuggugucu cgugaucuuu cucgcugagg ucgcagcguc acuugugguc 2640
cucuggcugc uugguaauac gcccuugcaa gacaaaggca auucuacaca cucaagaaac 2700
aauuccuaug ccgugauuau cacuucuaca agcucguauu acguguuuua caucuacgua 2760
ggaguggccg acacucugcu cgcgaugggu uucuuccgag gacucccacu cguucacacg 2820
cuuaucacug ucuccaagau ucuccaccau aagaugcuuc auagcguacu gcaggcuccc 2880
auguccaccu ugaauacgcu caaggcggga gguauuuuga aucgcuucuc aaaagauauu 2940
gcaauuuugg augaccuucu gccccugacg aucuucgacu ucauccaguu guugcugauc 3000
gugauugggg cuauugcagu agucgcuguc cuccagccuu acauuuuugu cgcgaccguu 3060
ccggugaucg uggcguuuau caugcugcgg gccuauuucu ugcagacguc acagcagcuu 3120
aagcaacugg agucugaagg gaggucgccu aucuuuacgc aucuugugac caguuugaag 3180
ggauugugga cguugcgcgc cuuuggcagg cagcccuacu uugaaacacu guuccacaaa 3240
gcgcugaauc uccauacggc aaauugguuu uuguauuuga guacccuccg augguuucag 3300
augcgcauug agaugauuuu ugugaucuuc uuuaucgcgg ugacuuuuau cuccaucuug 3360
accacgggag agggcgaggg acgggucggu auuauccuga cacucgccau gaacauuaug 3420
agcacuuugc agugggcagu gaacagcucg auugaugugg auagccugau gagguccguu 3480
ucgagggucu uuaaguucau cgacaugccg acggagggaa agcccacaaa aaguacgaaa 3540
cccuauaaga augggcaauu gaguaaggua augaucaucg agaacaguca cgugaagaag 3600
gaugacaucu ggccuagcgg gggucagaug accgugaagg accugacggc aaaauacacc 3660
gagggaggga acgcaauccu ugaaaacauc ucguucagca uuagccccgg ucagcgugug 3720
ggguugcucg ggaggaccgg gucaggaaaa ucgacguugc ugucggccuu cuugagacuu 3780
cugaauacag agggugagau ccagaucgac ggcguuucgu gggauagcau caccuugcag 3840
caguggcgga aagcguuugg aguaaucccc caaaaggucu uuaucuuuag cggaaccuuc 3900
cgaaagaauc ucgauccuua ugaacagugg ucagaucaag agauuuggaa agucgcggac 3960
gagguuggcc uucggagugu aaucgagcag uuuccgggaa aacucgacuu uguccuugua 4020
gaugggggau gcguccuguc gcaugggcac aagcagcuca ugugccuggc gcgauccguc 4080
cucucuaaag cgaaaauucu ucucuuggau gaaccuucgg cccaucugga cccgguaacg 4140
uaucagauca ucagaaggac acuuaagcag gcguuugccg acugcacggu gauucucugu 4200
gagcaucgua ucgaggccau gcucgaaugc cagcaauuuc uugucaucga agagaauaag 4260
guccgccagu acgacuccau ccagaagcug cuuaaugaga gaucauuguu ccggcaggcg 4320
auuucaccau ccgauagggu gaaacuuuuu ccacacagaa auucgucgaa gugcaagucc 4380
aaaccgcaga ucgcggccuu gaaagaagag acugaagaag aaguucaaga cacgcgucuu 4440
uaa 4443
<210> 4
<211> 1480
<212> PRT
<213> 智人
<400> 4
Met Gln Arg Ser Pro Leu Glu Lys Ala Ser Val Val Ser Lys Leu Phe
1 5 10 15
Phe Ser Trp Thr Arg Pro Ile Leu Arg Lys Gly Tyr Arg Gln Arg Leu
20 25 30
Glu Leu Ser Asp Ile Tyr Gln Ile Pro Ser Val Asp Ser Ala Asp Asn
35 40 45
Leu Ser Glu Lys Leu Glu Arg Glu Trp Asp Arg Glu Leu Ala Ser Lys
50 55 60
Lys Asn Pro Lys Leu Ile Asn Ala Leu Arg Arg Cys Phe Phe Trp Arg
65 70 75 80
Phe Met Phe Tyr Gly Ile Phe Leu Tyr Leu Gly Glu Val Thr Lys Ala
85 90 95
Val Gln Pro Leu Leu Leu Gly Arg Ile Ile Ala Ser Tyr Asp Pro Asp
100 105 110
Asn Lys Glu Glu Arg Ser Ile Ala Ile Tyr Leu Gly Ile Gly Leu Cys
115 120 125
Leu Leu Phe Ile Val Arg Thr Leu Leu Leu His Pro Ala Ile Phe Gly
130 135 140
Leu His His Ile Gly Met Gln Met Arg Ile Ala Met Phe Ser Leu Ile
145 150 155 160
Tyr Lys Lys Thr Leu Lys Leu Ser Ser Arg Val Leu Asp Lys Ile Ser
165 170 175
Ile Gly Gln Leu Val Ser Leu Leu Ser Asn Asn Leu Asn Lys Phe Asp
180 185 190
Glu Gly Leu Ala Leu Ala His Phe Val Trp Ile Ala Pro Leu Gln Val
195 200 205
Ala Leu Leu Met Gly Leu Ile Trp Glu Leu Leu Gln Ala Ser Ala Phe
210 215 220
Cys Gly Leu Gly Phe Leu Ile Val Leu Ala Leu Phe Gln Ala Gly Leu
225 230 235 240
Gly Arg Met Met Met Lys Tyr Arg Asp Gln Arg Ala Gly Lys Ile Ser
245 250 255
Glu Arg Leu Val Ile Thr Ser Glu Met Ile Glu Asn Ile Gln Ser Val
260 265 270
Lys Ala Tyr Cys Trp Glu Glu Ala Met Glu Lys Met Ile Glu Asn Leu
275 280 285
Arg Gln Thr Glu Leu Lys Leu Thr Arg Lys Ala Ala Tyr Val Arg Tyr
290 295 300
Phe Asn Ser Ser Ala Phe Phe Phe Ser Gly Phe Phe Val Val Phe Leu
305 310 315 320
Ser Val Leu Pro Tyr Ala Leu Ile Lys Gly Ile Ile Leu Arg Lys Ile
325 330 335
Phe Thr Thr Ile Ser Phe Cys Ile Val Leu Arg Met Ala Val Thr Arg
340 345 350
Gln Phe Pro Trp Ala Val Gln Thr Trp Tyr Asp Ser Leu Gly Ala Ile
355 360 365
Asn Lys Ile Gln Asp Phe Leu Gln Lys Gln Glu Tyr Lys Thr Leu Glu
370 375 380
Tyr Asn Leu Thr Thr Thr Glu Val Val Met Glu Asn Val Thr Ala Phe
385 390 395 400
Trp Glu Glu Gly Phe Gly Glu Leu Phe Glu Lys Ala Lys Gln Asn Asn
405 410 415
Asn Asn Arg Lys Thr Ser Asn Gly Asp Asp Ser Leu Phe Phe Ser Asn
420 425 430
Phe Ser Leu Leu Gly Thr Pro Val Leu Lys Asp Ile Asn Phe Lys Ile
435 440 445
Glu Arg Gly Gln Leu Leu Ala Val Ala Gly Ser Thr Gly Ala Gly Lys
450 455 460
Thr Ser Leu Leu Met Val Ile Met Gly Glu Leu Glu Pro Ser Glu Gly
465 470 475 480
Lys Ile Lys His Ser Gly Arg Ile Ser Phe Cys Ser Gln Phe Ser Trp
485 490 495
Ile Met Pro Gly Thr Ile Lys Glu Asn Ile Ile Phe Gly Val Ser Tyr
500 505 510
Asp Glu Tyr Arg Tyr Arg Ser Val Ile Lys Ala Cys Gln Leu Glu Glu
515 520 525
Asp Ile Ser Lys Phe Ala Glu Lys Asp Asn Ile Val Leu Gly Glu Gly
530 535 540
Gly Ile Thr Leu Ser Gly Gly Gln Arg Ala Arg Ile Ser Leu Ala Arg
545 550 555 560
Ala Val Tyr Lys Asp Ala Asp Leu Tyr Leu Leu Asp Ser Pro Phe Gly
565 570 575
Tyr Leu Asp Val Leu Thr Glu Lys Glu Ile Phe Glu Ser Cys Val Cys
580 585 590
Lys Leu Met Ala Asn Lys Thr Arg Ile Leu Val Thr Ser Lys Met Glu
595 600 605
His Leu Lys Lys Ala Asp Lys Ile Leu Ile Leu His Glu Gly Ser Ser
610 615 620
Tyr Phe Tyr Gly Thr Phe Ser Glu Leu Gln Asn Leu Gln Pro Asp Phe
625 630 635 640
Ser Ser Lys Leu Met Gly Cys Asp Ser Phe Asp Gln Phe Ser Ala Glu
645 650 655
Arg Arg Asn Ser Ile Leu Thr Glu Thr Leu His Arg Phe Ser Leu Glu
660 665 670
Gly Asp Ala Pro Val Ser Trp Thr Glu Thr Lys Lys Gln Ser Phe Lys
675 680 685
Gln Thr Gly Glu Phe Gly Glu Lys Arg Lys Asn Ser Ile Leu Asn Pro
690 695 700
Ile Asn Ser Ile Arg Lys Phe Ser Ile Val Gln Lys Thr Pro Leu Gln
705 710 715 720
Met Asn Gly Ile Glu Glu Asp Ser Asp Glu Pro Leu Glu Arg Arg Leu
725 730 735
Ser Leu Val Pro Asp Ser Glu Gln Gly Glu Ala Ile Leu Pro Arg Ile
740 745 750
Ser Val Ile Ser Thr Gly Pro Thr Leu Gln Ala Arg Arg Arg Gln Ser
755 760 765
Val Leu Asn Leu Met Thr His Ser Val Asn Gln Gly Gln Asn Ile His
770 775 780
Arg Lys Thr Thr Ala Ser Thr Arg Lys Val Ser Leu Ala Pro Gln Ala
785 790 795 800
Asn Leu Thr Glu Leu Asp Ile Tyr Ser Arg Arg Leu Ser Gln Glu Thr
805 810 815
Gly Leu Glu Ile Ser Glu Glu Ile Asn Glu Glu Asp Leu Lys Glu Cys
820 825 830
Phe Phe Asp Asp Met Glu Ser Ile Pro Ala Val Thr Thr Trp Asn Thr
835 840 845
Tyr Leu Arg Tyr Ile Thr Val His Lys Ser Leu Ile Phe Val Leu Ile
850 855 860
Trp Cys Leu Val Ile Phe Leu Ala Glu Val Ala Ala Ser Leu Val Val
865 870 875 880
Leu Trp Leu Leu Gly Asn Thr Pro Leu Gln Asp Lys Gly Asn Ser Thr
885 890 895
His Ser Arg Asn Asn Ser Tyr Ala Val Ile Ile Thr Ser Thr Ser Ser
900 905 910
Tyr Tyr Val Phe Tyr Ile Tyr Val Gly Val Ala Asp Thr Leu Leu Ala
915 920 925
Met Gly Phe Phe Arg Gly Leu Pro Leu Val His Thr Leu Ile Thr Val
930 935 940
Ser Lys Ile Leu His His Lys Met Leu His Ser Val Leu Gln Ala Pro
945 950 955 960
Met Ser Thr Leu Asn Thr Leu Lys Ala Gly Gly Ile Leu Asn Arg Phe
965 970 975
Ser Lys Asp Ile Ala Ile Leu Asp Asp Leu Leu Pro Leu Thr Ile Phe
980 985 990
Asp Phe Ile Gln Leu Leu Leu Ile Val Ile Gly Ala Ile Ala Val Val
995 1000 1005
Ala Val Leu Gln Pro Tyr Ile Phe Val Ala Thr Val Pro Val Ile
1010 1015 1020
Val Ala Phe Ile Met Leu Arg Ala Tyr Phe Leu Gln Thr Ser Gln
1025 1030 1035
Gln Leu Lys Gln Leu Glu Ser Glu Gly Arg Ser Pro Ile Phe Thr
1040 1045 1050
His Leu Val Thr Ser Leu Lys Gly Leu Trp Thr Leu Arg Ala Phe
1055 1060 1065
Gly Arg Gln Pro Tyr Phe Glu Thr Leu Phe His Lys Ala Leu Asn
1070 1075 1080
Leu His Thr Ala Asn Trp Phe Leu Tyr Leu Ser Thr Leu Arg Trp
1085 1090 1095
Phe Gln Met Arg Ile Glu Met Ile Phe Val Ile Phe Phe Ile Ala
1100 1105 1110
Val Thr Phe Ile Ser Ile Leu Thr Thr Gly Glu Gly Glu Gly Arg
1115 1120 1125
Val Gly Ile Ile Leu Thr Leu Ala Met Asn Ile Met Ser Thr Leu
1130 1135 1140
Gln Trp Ala Val Asn Ser Ser Ile Asp Val Asp Ser Leu Met Arg
1145 1150 1155
Ser Val Ser Arg Val Phe Lys Phe Ile Asp Met Pro Thr Glu Gly
1160 1165 1170
Lys Pro Thr Lys Ser Thr Lys Pro Tyr Lys Asn Gly Gln Leu Ser
1175 1180 1185
Lys Val Met Ile Ile Glu Asn Ser His Val Lys Lys Asp Asp Ile
1190 1195 1200
Trp Pro Ser Gly Gly Gln Met Thr Val Lys Asp Leu Thr Ala Lys
1205 1210 1215
Tyr Thr Glu Gly Gly Asn Ala Ile Leu Glu Asn Ile Ser Phe Ser
1220 1225 1230
Ile Ser Pro Gly Gln Arg Val Gly Leu Leu Gly Arg Thr Gly Ser
1235 1240 1245
Gly Lys Ser Thr Leu Leu Ser Ala Phe Leu Arg Leu Leu Asn Thr
1250 1255 1260
Glu Gly Glu Ile Gln Ile Asp Gly Val Ser Trp Asp Ser Ile Thr
1265 1270 1275
Leu Gln Gln Trp Arg Lys Ala Phe Gly Val Ile Pro Gln Lys Val
1280 1285 1290
Phe Ile Phe Ser Gly Thr Phe Arg Lys Asn Leu Asp Pro Tyr Glu
1295 1300 1305
Gln Trp Ser Asp Gln Glu Ile Trp Lys Val Ala Asp Glu Val Gly
1310 1315 1320
Leu Arg Ser Val Ile Glu Gln Phe Pro Gly Lys Leu Asp Phe Val
1325 1330 1335
Leu Val Asp Gly Gly Cys Val Leu Ser His Gly His Lys Gln Leu
1340 1345 1350
Met Cys Leu Ala Arg Ser Val Leu Ser Lys Ala Lys Ile Leu Leu
1355 1360 1365
Leu Asp Glu Pro Ser Ala His Leu Asp Pro Val Thr Tyr Gln Ile
1370 1375 1380
Ile Arg Arg Thr Leu Lys Gln Ala Phe Ala Asp Cys Thr Val Ile
1385 1390 1395
Leu Cys Glu His Arg Ile Glu Ala Met Leu Glu Cys Gln Gln Phe
1400 1405 1410
Leu Val Ile Glu Glu Asn Lys Val Arg Gln Tyr Asp Ser Ile Gln
1415 1420 1425
Lys Leu Leu Asn Glu Arg Ser Leu Phe Arg Gln Ala Ile Ser Pro
1430 1435 1440
Ser Asp Arg Val Lys Leu Phe Pro His Arg Asn Ser Ser Lys Cys
1445 1450 1455
Lys Ser Lys Pro Gln Ile Ala Ala Leu Lys Glu Glu Thr Glu Glu
1460 1465 1470
Glu Val Gln Asp Thr Arg Leu
1475 1480
<210> 5
<211> 140
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 5
ggacagaucg ccuggagacg ccauccacgc uguuuugacc uccauagaag acaccgggac 60
cgauccagcc uccgcggccg ggaacggugc auuggaacgc ggauuccccg ugccaagagu 120
gacucaccgu ccuugacacg 140
<210> 6
<211> 105
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 6
cggguggcau cccugugacc ccuccccagu gccucuccug gcccuggaag uugccacucc 60
agugcccacc agccuugucc uaauaaaauu aaguugcauc aagcu 105
<210> 7
<211> 105
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 7
ggguggcauc ccugugaccc cuccccagug ccucuccugg cccuggaagu ugccacucca 60
gugcccacca gccuuguccu aauaaaauua aguugcauca aagcu 105
<210> 8
<211> 4688
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 8
ggacagaucg ccuggagacg ccauccacgc uguuuugacc uccauagaag acaccgggac 60
cgauccagcc uccgcggccg ggaacggugc auuggaacgc ggauuccccg ugccaagagu 120
gacucaccgu ccuugacacg augcaacgcu cuccucuuga aaaggccucg guggugucca 180
agcucuucuu cucguggacu agacccaucc ugagaaaggg guacagacag cgcuuggagc 240
uguccgauau cuaucaaauc ccuuccgugg acuccgcgga caaccugucc gagaagcucg 300
agagagaaug ggacagagaa cucgccucaa agaagaaccc gaagcugauu aaugcgcuua 360
ggcggugcuu uuucuggcgg uucauguucu acggcaucuu ccucuaccug ggagagguca 420
ccaaggccgu gcagccccug uugcugggac ggauuauugc cuccuacgac cccgacaaca 480
aggaagaaag aagcaucgcu aucuacuugg gcaucggucu gugccugcuu uucaucgucc 540
ggacccucuu guugcauccu gcuauuuucg gccugcauca cauuggcaug cagaugagaa 600
uugccauguu uucccugauc uacaagaaaa cucugaagcu cucgagccgc gugcuugaca 660
agauuuccau cggccagcuc gugucccugc ucuccaacaa ucugaacaag uucgacgagg 720
gccucgcccu ggcccacuuc guguggaucg ccccucugca aguggcgcuu cugaugggcc 780
ugaucuggga gcugcugcaa gccucggcau ucugugggcu uggauuccug aucgugcugg 840
cacuguucca ggccggacug gggcggauga ugaugaagua cagggaccag agagccggaa 900
agauuuccga acggcuggug aucacuucgg aaaugaucga aaacauccag ucagugaagg 960
ccuacugcug ggaagaggcc auggaaaaga ugauugaaaa ccuccggcaa accgagcuga 1020
agcugacccg caaggccgcu uacgugcgcu auuucaacuc guccgcuuuc uucuucuccg 1080
gguucuucgu gguguuucuc uccgugcucc ccuacgcccu gauuaaggga aucauccuca 1140
ggaagaucuu caccaccauu uccuucugua ucgugcuccg cauggccgug acccggcagu 1200
ucccaugggc cgugcagacu ugguacgacu cccugggagc cauuaacaag auccaggacu 1260
uccuucaaaa gcaggaguac aagacccucg aguacaaccu gacuacuacc gaggucguga 1320
uggaaaacgu caccgccuuu ugggaggagg gauuuggcga acuguucgag aaggccaagc 1380
agaacaacaa caaccgcaag accucgaacg gugacgacuc ccucuucuuu ucaaacuuca 1440
gccugcucgg gacgcccgug cugaaggaca uuaacuucaa gaucgaaaga ggacagcucc 1500
uggcgguggc cggaucgacc ggagccggaa agacuucccu gcugauggug aucaugggag 1560
agcuugaacc uagcgaggga aagaucaagc acuccggccg caucagcuuc uguagccagu 1620
uuuccuggau caugcccgga accauuaagg aaaacaucau cuucggcgug uccuacgaug 1680
aauaccgcua ccgguccgug aucaaagccu gccagcugga agaggauauu ucaaaguucg 1740
cggagaaaga uaacaucgug cugggcgaag gggguauuac cuugucgggg ggccagcggg 1800
cuagaaucuc gcuggccaga gccguguaua aggacgccga ccuguaucuc cuggacuccc 1860
ccuucggaua ccuggacguc cugaccgaaa aggagaucuu cgaaucgugc gugugcaagc 1920
ugauggcuaa caagacucgc auccucguga ccuccaaaau ggagcaccug aagaaggcag 1980
acaagauucu gauucugcau gagggguccu ccuacuuuua cggcaccuuc ucggaguugc 2040
agaacuugca gcccgacuuc ucaucgaagc ugauggguug cgacagcuuc gaccaguucu 2100
ccgccgaaag aaggaacucg auccugacgg aaaccuugca ccgcuucucu uuggaaggcg 2160
acgccccugu gucauggacc gagacuaaga agcagagcuu caagcagacc ggggaauucg 2220
gcgaaaagag gaagaacagc aucuugaacc ccauuaacuc cauccgcaag uucucaaucg 2280
ugcaaaagac gccacugcag augaacggca uugaggagga cuccgacgaa ccccuugaga 2340
ggcgccuguc ccuggugccg gacagcgagc agggagaagc cauccugccu cggauuuccg 2400
ugaucuccac ugguccgacg cuccaagccc ggcggcggca guccgugcug aaccugauga 2460
cccacagcgu gaaccagggc caaaacauuc accgcaagac uaccgcaucc acccggaaag 2520
ugucccuggc accucaagcg aaucuuaccg agcucgacau cuacucccgg agacugucgc 2580
aggaaaccgg gcucgaaauu uccgaagaaa ucaacgagga ggaucugaaa gagugcuucu 2640
ucgacgauau ggagucgaua cccgccguga cgacuuggaa cacuuaucug cgguacauca 2700
cugugcacaa gucauugauc uucgugcuga uuuggugccu ggugauuuuc cuggccgagg 2760
ucgcggccuc acugguggug cucuggcugu ugggaaacac gccucugcaa gacaagggaa 2820
acuccacgca cucgagaaac aacagcuaug ccgugauuau cacuuccacc uccucuuauu 2880
acguguucua caucuacguc ggaguggcgg auacccugcu cgcgaugggu uucuucagag 2940
gacugccgcu gguccacacc uugaucaccg ucagcaagau ucuucaccac aagauguugc 3000
auagcgugcu gcaggccccc auguccaccc ucaacacucu gaaggccgga ggcauucuga 3060
acagauucuc caaggacauc gcuauccugg acgaucuccu gccgcuuacc aucuuugacu 3120
ucauccagcu gcugcugauc gugauuggag caaucgcagu gguggcggug cugcagccuu 3180
acauuuucgu ggccacugug ccggucauug uggcguucau caugcugcgg gccuacuucc 3240
uccaaaccag ccagcagcug aagcaacugg aauccgaggg acgauccccc aucuucacuc 3300
accuugugac gucguugaag ggacugugga cccuccgggc uuucggacgg cagcccuacu 3360
ucgaaacccu cuuccacaag gcccugaacc uccacaccgc caauugguuc cuguaccugu 3420
ccacccugcg gugguuccag augcgcaucg agaugauuuu cgucaucuuc uucaucgcgg 3480
ucacauucau cagcauccug acuaccggag agggagaggg acgggucgga auaauccuga 3540
cccucgccau gaacauuaug agcacccugc agugggcagu gaacagcucg aucgacgugg 3600
acagccugau gcgaagcguc agccgcgugu ucaaguucau cgacaugccu acugagggaa 3660
aacccacuaa guccacuaag cccuacaaaa auggccagcu gagcaagguc augaucaucg 3720
aaaacuccca cgugaagaag gacgauauuu ggcccuccgg aggucaaaug accgugaagg 3780
accugaccgc aaaguacacc gagggaggaa acgccauucu cgaaaacauc agcuucucca 3840
uuucgccggg acagcggguc ggccuucucg ggcggaccgg uuccgggaag ucaacucugc 3900
ugucggcuuu ccuccggcug cugaauaccg agggggaaau ccaaauugac ggcgugucuu 3960
gggauuccau uacucugcag caguggcgga aggccuucgg cgugaucccc cagaaggugu 4020
ucaucuucuc ggguaccuuc cggaagaacc uggauccuua cgagcagugg agcgaccaag 4080
aaaucuggaa ggucgccgac gaggucggcc ugcgcuccgu gauugaacaa uuuccuggaa 4140
agcuggacuu cgugcucguc gacgggggau guguccuguc gcacggacau aagcagcuca 4200
ugugccucgc acgguccgug cucuccaagg ccaagauucu gcugcuggac gaaccuucgg 4260
cccaccugga uccggucacc uaccagauca ucaggaggac ccugaagcag gccuuugccg 4320
auugcaccgu gauucucugc gagcaccgca ucgaggccau gcuggagugc cagcaguucc 4380
uggucaucga ggagaacaag guccgccaau acgacuccau ucaaaagcuc cucaacgagc 4440
ggucgcuguu cagacaagcu auuucaccgu ccgauagagu gaagcucuuc ccgcaucgga 4500
acagcucaaa gugcaaaucg aagccgcaga ucgcagccuu gaaggaagag acugaggaag 4560
aggugcagga cacccggcuu uaacgggugg caucccugug accccucccc agugccucuc 4620
cuggcccugg aaguugccac uccagugccc accagccuug uccuaauaaa auuaaguugc 4680
aucaagcu 4688
<210> 9
<211> 4688
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 9
ggacagaucg ccuggagacg ccauccacgc uguuuugacc uccauagaag acaccgggac 60
cgauccagcc uccgcggccg ggaacggugc auuggaacgc ggauuccccg ugccaagagu 120
gacucaccgu ccuugacacg augcaacgcu cuccucuuga aaaggccucg guggugucca 180
agcucuucuu cucguggacu agacccaucc ugagaaaggg guacagacag cgcuuggagc 240
uguccgauau cuaucaaauc ccuuccgugg acuccgcgga caaccugucc gagaagcucg 300
agagagaaug ggacagagaa cucgccucaa agaagaaccc gaagcugauu aaugcgcuua 360
ggcggugcuu uuucuggcgg uucauguucu acggcaucuu ccucuaccug ggagagguca 420
ccaaggccgu gcagccccug uugcugggac ggauuauugc cuccuacgac cccgacaaca 480
aggaagaaag aagcaucgcu aucuacuugg gcaucggucu gugccugcuu uucaucgucc 540
ggacccucuu guugcauccu gcuauuuucg gccugcauca cauuggcaug cagaugagaa 600
uugccauguu uucccugauc uacaagaaaa cucugaagcu cucgagccgc gugcuugaca 660
agauuuccau cggccagcuc gugucccugc ucuccaacaa ucugaacaag uucgacgagg 720
gccucgcccu ggcccacuuc guguggaucg ccccucugca aguggcgcuu cugaugggcc 780
ugaucuggga gcugcugcaa gccucggcau ucugugggcu uggauuccug aucgugcugg 840
cacuguucca ggccggacug gggcggauga ugaugaagua cagggaccag agagccggaa 900
agauuuccga acggcuggug aucacuucgg aaaugaucga aaacauccag ucagugaagg 960
ccuacugcug ggaagaggcc auggaaaaga ugauugaaaa ccuccggcaa accgagcuga 1020
agcugacccg caaggccgcu uacgugcgcu auuucaacuc guccgcuuuc uucuucuccg 1080
gguucuucgu gguguuucuc uccgugcucc ccuacgcccu gauuaaggga aucauccuca 1140
ggaagaucuu caccaccauu uccuucugua ucgugcuccg cauggccgug acccggcagu 1200
ucccaugggc cgugcagacu ugguacgacu cccugggagc cauuaacaag auccaggacu 1260
uccuucaaaa gcaggaguac aagacccucg aguacaaccu gacuacuacc gaggucguga 1320
uggaaaacgu caccgccuuu ugggaggagg gauuuggcga acuguucgag aaggccaagc 1380
agaacaacaa caaccgcaag accucgaacg gugacgacuc ccucuucuuu ucaaacuuca 1440
gccugcucgg gacgcccgug cugaaggaca uuaacuucaa gaucgaaaga ggacagcucc 1500
uggcgguggc cggaucgacc ggagccggaa agacuucccu gcugauggug aucaugggag 1560
agcuugaacc uagcgaggga aagaucaagc acuccggccg caucagcuuc uguagccagu 1620
uuuccuggau caugcccgga accauuaagg aaaacaucau cuucggcgug uccuacgaug 1680
aauaccgcua ccgguccgug aucaaagccu gccagcugga agaggauauu ucaaaguucg 1740
cggagaaaga uaacaucgug cugggcgaag gggguauuac cuugucgggg ggccagcggg 1800
cuagaaucuc gcuggccaga gccguguaua aggacgccga ccuguaucuc cuggacuccc 1860
ccuucggaua ccuggacguc cugaccgaaa aggagaucuu cgaaucgugc gugugcaagc 1920
ugauggcuaa caagacucgc auccucguga ccuccaaaau ggagcaccug aagaaggcag 1980
acaagauucu gauucugcau gagggguccu ccuacuuuua cggcaccuuc ucggaguugc 2040
agaacuugca gcccgacuuc ucaucgaagc ugauggguug cgacagcuuc gaccaguucu 2100
ccgccgaaag aaggaacucg auccugacgg aaaccuugca ccgcuucucu uuggaaggcg 2160
acgccccugu gucauggacc gagacuaaga agcagagcuu caagcagacc ggggaauucg 2220
gcgaaaagag gaagaacagc aucuugaacc ccauuaacuc cauccgcaag uucucaaucg 2280
ugcaaaagac gccacugcag augaacggca uugaggagga cuccgacgaa ccccuugaga 2340
ggcgccuguc ccuggugccg gacagcgagc agggagaagc cauccugccu cggauuuccg 2400
ugaucuccac ugguccgacg cuccaagccc ggcggcggca guccgugcug aaccugauga 2460
cccacagcgu gaaccagggc caaaacauuc accgcaagac uaccgcaucc acccggaaag 2520
ugucccuggc accucaagcg aaucuuaccg agcucgacau cuacucccgg agacugucgc 2580
aggaaaccgg gcucgaaauu uccgaagaaa ucaacgagga ggaucugaaa gagugcuucu 2640
ucgacgauau ggagucgaua cccgccguga cgacuuggaa cacuuaucug cgguacauca 2700
cugugcacaa gucauugauc uucgugcuga uuuggugccu ggugauuuuc cuggccgagg 2760
ucgcggccuc acugguggug cucuggcugu ugggaaacac gccucugcaa gacaagggaa 2820
acuccacgca cucgagaaac aacagcuaug ccgugauuau cacuuccacc uccucuuauu 2880
acguguucua caucuacguc ggaguggcgg auacccugcu cgcgaugggu uucuucagag 2940
gacugccgcu gguccacacc uugaucaccg ucagcaagau ucuucaccac aagauguugc 3000
auagcgugcu gcaggccccc auguccaccc ucaacacucu gaaggccgga ggcauucuga 3060
acagauucuc caaggacauc gcuauccugg acgaucuccu gccgcuuacc aucuuugacu 3120
ucauccagcu gcugcugauc gugauuggag caaucgcagu gguggcggug cugcagccuu 3180
acauuuucgu ggccacugug ccggucauug uggcguucau caugcugcgg gccuacuucc 3240
uccaaaccag ccagcagcug aagcaacugg aauccgaggg acgauccccc aucuucacuc 3300
accuugugac gucguugaag ggacugugga cccuccgggc uuucggacgg cagcccuacu 3360
ucgaaacccu cuuccacaag gcccugaacc uccacaccgc caauugguuc cuguaccugu 3420
ccacccugcg gugguuccag augcgcaucg agaugauuuu cgucaucuuc uucaucgcgg 3480
ucacauucau cagcauccug acuaccggag agggagaggg acgggucgga auaauccuga 3540
cccucgccau gaacauuaug agcacccugc agugggcagu gaacagcucg aucgacgugg 3600
acagccugau gcgaagcguc agccgcgugu ucaaguucau cgacaugccu acugagggaa 3660
aacccacuaa guccacuaag cccuacaaaa auggccagcu gagcaagguc augaucaucg 3720
aaaacuccca cgugaagaag gacgauauuu ggcccuccgg aggucaaaug accgugaagg 3780
accugaccgc aaaguacacc gagggaggaa acgccauucu cgaaaacauc agcuucucca 3840
uuucgccggg acagcggguc ggccuucucg ggcggaccgg uuccgggaag ucaacucugc 3900
ugucggcuuu ccuccggcug cugaauaccg agggggaaau ccaaauugac ggcgugucuu 3960
gggauuccau uacucugcag caguggcgga aggccuucgg cgugaucccc cagaaggugu 4020
ucaucuucuc ggguaccuuc cggaagaacc uggauccuua cgagcagugg agcgaccaag 4080
aaaucuggaa ggucgccgac gaggucggcc ugcgcuccgu gauugaacaa uuuccuggaa 4140
agcuggacuu cgugcucguc gacgggggau guguccuguc gcacggacau aagcagcuca 4200
ugugccucgc acgguccgug cucuccaagg ccaagauucu gcugcuggac gaaccuucgg 4260
cccaccugga uccggucacc uaccagauca ucaggaggac ccugaagcag gccuuugccg 4320
auugcaccgu gauucucugc gagcaccgca ucgaggccau gcuggagugc cagcaguucc 4380
uggucaucga ggagaacaag guccgccaau acgacuccau ucaaaagcuc cucaacgagc 4440
ggucgcuguu cagacaagcu auuucaccgu ccgauagagu gaagcucuuc ccgcaucgga 4500
acagcucaaa gugcaaaucg aagccgcaga ucgcagccuu gaaggaagag acugaggaag 4560
aggugcagga cacccggcuu uaaggguggc aucccuguga ccccucccca gugccucucc 4620
uggcccugga aguugccacu ccagugccca ccagccuugu ccuaauaaaa uuaaguugca 4680
ucaaagcu 4688
<210> 10
<211> 4688
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 10
ggacagaucg ccuggagacg ccauccacgc uguuuugacc uccauagaag acaccgggac 60
cgauccagcc uccgcggccg ggaacggugc auuggaacgc ggauuccccg ugccaagagu 120
gacucaccgu ccuugacacg augcagcggu ccccgcucga aaaggccagu gucgugucca 180
aacucuucuu cucauggacu cggccuaucc uuagaaaggg guaucggcag aggcuugagu 240
ugucugacau cuaccagauc cccucgguag auucggcgga uaaccucucg gagaagcucg 300
aacgggaaug ggaccgcgaa cucgcgucua agaaaaaccc gaagcucauc aacgcacuga 360
gaaggugcuu cuucuggcgg uucauguucu acgguaucuu cuuguaucuc ggggagguca 420
caaaagcagu ccaaccccug uuguuggguc gcauuaucgc cucguacgac cccgauaaca 480
aagaagaacg gagcaucgcg aucuaccucg ggaucggacu guguuugcuu uucaucguca 540
gaacacuuuu guugcaucca gcaaucuucg gccuccauca caucgguaug cagaugcgaa 600
ucgcuauguu uagcuugauc uacaaaaaga cacugaaacu cucgucgcgg guguuggaua 660
agauuuccau cggucaguug gugucccugc uuaguaauaa ccucaacaaa uucgaugagg 720
gacuggcgcu ggcacauuuc guguggauug ccccguugca agucgcccuu uugaugggcc 780
uuauuuggga gcuguugcag gcaucugccu uuuguggccu gggauuucug auuguguugg 840
cauuguuuca ggcugggcuu gggcggauga ugaugaagua ucgcgaccag agagcgggua 900
aaaucucgga aagacucguc aucacuucgg aaaugaucga aaacauccag ucggucaaag 960
ccuauugcug ggaagaagcu auggagaaga ugauugaaaa ccuccgccaa acugagcuga 1020
aacugacccg caaggcggcg uauguccggu auuucaauuc gucagcguuc uucuuuuccg 1080
gguucuucgu ugucuuucuc ucgguuuugc cuuaugccuu gauuaagggg auuauccucc 1140
gcaagauuuu caccacgauu ucguucugca uuguauugcg cauggcagug acacggcaau 1200
uuccgugggc cgugcagaca ugguaugacu cgcuuggagc gaucaacaaa auccaagacu 1260
ucuugcaaaa gcaagaguac aagacccugg aguacaaucu uacuacuacg gagguaguaa 1320
uggagaaugu gacggcuuuu ugggaagagg guuuuggaga acuguuugag aaagcaaagc 1380
agaauaacaa caaccgcaag accucaaaug gggacgauuc ccuguuuuuc ucgaacuucu 1440
cccugcucgg aacacccgug uugaaggaca ucaauuucaa gauugagagg ggacagcuuc 1500
ucgcgguagc gggaagcacu ggugcgggaa aaacuagccu cuugauggug auuauggggg 1560
agcuugagcc cagcgagggg aagauuaaac acuccgggcg uaucucauuc uguagccagu 1620
uuucauggau caugcccgga accauuaaag agaacaucau uuucggagua uccuaugaug 1680
aguaccgaua cagaucgguc auuaaggcgu gccaguugga agaggacauu ucuaaguucg 1740
ccgagaagga uaacaucguc uugggagaag gggguauuac auugucggga gggcagcgag 1800
cgcggaucag ccucgcgaga gcgguauaca aagaugcaga uuuguaucug cuugauucac 1860
cguuuggaua ccucgacgua uugacagaaa aagaaaucuu cgagucgugc guguguaaac 1920
uuauggcuaa uaagacgaga auccugguga caucaaaaau ggaacaccuu aagaaggcgg 1980
acaagauccu gauccuccac gaaggaucgu ccuacuuuua cggcacuuuc ucagaguugc 2040
aaaacuugca gccggacuuc ucaagcaaac ucauggggug ugacucauuc gaccaguuca 2100
gcgcggaacg gcggaacucg aucuugacgg aaacgcugca ccgauucucg cuugagggug 2160
augccccggu aucguggacc gagacaaaga agcagucguu uaagcagaca ggagaauuug 2220
gugagaaaag aaagaacagu aucuugaauc cuauuaacuc aauucgcaag uucucaaucg 2280
uccagaaaac uccacugcag augaauggaa uugaagagga uucggacgaa ccccuggagc 2340
gcaggcuuag ccucgugccg gauucagagc aaggggaggc cauucuuccc cggauuucgg 2400
ugauuucaac cggaccuaca cuucaggcga ggcgaaggca auccgugcuc aaccucauga 2460
cgcauucggu aaaccagggg caaaacauuc accgcaaaac gacggccuca acgagaaaag 2520
ugucacuugc accccaggcg aauuugacug aacucgacau cuacagccgu aggcuuucgc 2580
aagaaaccgg acuugagauc agcgaagaaa ucaaugaaga agauuugaaa gaguguuucu 2640
uugaugacau ggaaucaauc ccagcgguga caacguggaa cacauacuug cguuacauca 2700
cggugcacaa guccuugauu uucguccuca ucuggugucu cgugaucuuu cucgcugagg 2760
ucgcagcguc acuugugguc cucuggcugc uugguaauac gcccuugcaa gacaaaggca 2820
auucuacaca cucaagaaac aauuccuaug ccgugauuau cacuucuaca agcucguauu 2880
acguguuuua caucuacgua ggaguggccg acacucugcu cgcgaugggu uucuuccgag 2940
gacucccacu cguucacacg cuuaucacug ucuccaagau ucuccaccau aagaugcuuc 3000
auagcguacu gcaggcuccc auguccaccu ugaauacgcu caaggcggga gguauuuuga 3060
aucgcuucuc aaaagauauu gcaauuuugg augaccuucu gccccugacg aucuucgacu 3120
ucauccaguu guugcugauc gugauugggg cuauugcagu agucgcuguc cuccagccuu 3180
acauuuuugu cgcgaccguu ccggugaucg uggcguuuau caugcugcgg gccuauuucu 3240
ugcagacguc acagcagcuu aagcaacugg agucugaagg gaggucgccu aucuuuacgc 3300
aucuugugac caguuugaag ggauugugga cguugcgcgc cuuuggcagg cagcccuacu 3360
uugaaacacu guuccacaaa gcgcugaauc uccauacggc aaauugguuu uuguauuuga 3420
guacccuccg augguuucag augcgcauug agaugauuuu ugugaucuuc uuuaucgcgg 3480
ugacuuuuau cuccaucuug accacgggag agggcgaggg acgggucggu auuauccuga 3540
cacucgccau gaacauuaug agcacuuugc agugggcagu gaacagcucg auugaugugg 3600
auagccugau gagguccguu ucgagggucu uuaaguucau cgacaugccg acggagggaa 3660
agcccacaaa aaguacgaaa cccuauaaga augggcaauu gaguaaggua augaucaucg 3720
agaacaguca cgugaagaag gaugacaucu ggccuagcgg gggucagaug accgugaagg 3780
accugacggc aaaauacacc gagggaggga acgcaauccu ugaaaacauc ucguucagca 3840
uuagccccgg ucagcgugug ggguugcucg ggaggaccgg gucaggaaaa ucgacguugc 3900
ugucggccuu cuugagacuu cugaauacag agggugagau ccagaucgac ggcguuucgu 3960
gggauagcau caccuugcag caguggcgga aagcguuugg aguaaucccc caaaaggucu 4020
uuaucuuuag cggaaccuuc cgaaagaauc ucgauccuua ugaacagugg ucagaucaag 4080
agauuuggaa agucgcggac gagguuggcc uucggagugu aaucgagcag uuuccgggaa 4140
aacucgacuu uguccuugua gaugggggau gcguccuguc gcaugggcac aagcagcuca 4200
ugugccuggc gcgauccguc cucucuaaag cgaaaauucu ucucuuggau gaaccuucgg 4260
cccaucugga cccgguaacg uaucagauca ucagaaggac acuuaagcag gcguuugccg 4320
acugcacggu gauucucugu gagcaucgua ucgaggccau gcucgaaugc cagcaauuuc 4380
uugucaucga agagaauaag guccgccagu acgacuccau ccagaagcug cuuaaugaga 4440
gaucauuguu ccggcaggcg auuucaccau ccgauagggu gaaacuuuuu ccacacagaa 4500
auucgucgaa gugcaagucc aaaccgcaga ucgcggccuu gaaagaagag acugaagaag 4560
aaguucaaga cacgcgucuu uaacgggugg caucccugug accccucccc agugccucuc 4620
cuggcccugg aaguugccac uccagugccc accagccuug uccuaauaaa auuaaguugc 4680
aucaagcu 4688
<210> 11
<211> 4688
<212> RNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸
<400> 11
ggacagaucg ccuggagacg ccauccacgc uguuuugacc uccauagaag acaccgggac 60
cgauccagcc uccgcggccg ggaacggugc auuggaacgc ggauuccccg ugccaagagu 120
gacucaccgu ccuugacacg augcagcggu ccccgcucga aaaggccagu gucgugucca 180
aacucuucuu cucauggacu cggccuaucc uuagaaaggg guaucggcag aggcuugagu 240
ugucugacau cuaccagauc cccucgguag auucggcgga uaaccucucg gagaagcucg 300
aacgggaaug ggaccgcgaa cucgcgucua agaaaaaccc gaagcucauc aacgcacuga 360
gaaggugcuu cuucuggcgg uucauguucu acgguaucuu cuuguaucuc ggggagguca 420
caaaagcagu ccaaccccug uuguuggguc gcauuaucgc cucguacgac cccgauaaca 480
aagaagaacg gagcaucgcg aucuaccucg ggaucggacu guguuugcuu uucaucguca 540
gaacacuuuu guugcaucca gcaaucuucg gccuccauca caucgguaug cagaugcgaa 600
ucgcuauguu uagcuugauc uacaaaaaga cacugaaacu cucgucgcgg guguuggaua 660
agauuuccau cggucaguug gugucccugc uuaguaauaa ccucaacaaa uucgaugagg 720
gacuggcgcu ggcacauuuc guguggauug ccccguugca agucgcccuu uugaugggcc 780
uuauuuggga gcuguugcag gcaucugccu uuuguggccu gggauuucug auuguguugg 840
cauuguuuca ggcugggcuu gggcggauga ugaugaagua ucgcgaccag agagcgggua 900
aaaucucgga aagacucguc aucacuucgg aaaugaucga aaacauccag ucggucaaag 960
ccuauugcug ggaagaagcu auggagaaga ugauugaaaa ccuccgccaa acugagcuga 1020
aacugacccg caaggcggcg uauguccggu auuucaauuc gucagcguuc uucuuuuccg 1080
gguucuucgu ugucuuucuc ucgguuuugc cuuaugccuu gauuaagggg auuauccucc 1140
gcaagauuuu caccacgauu ucguucugca uuguauugcg cauggcagug acacggcaau 1200
uuccgugggc cgugcagaca ugguaugacu cgcuuggagc gaucaacaaa auccaagacu 1260
ucuugcaaaa gcaagaguac aagacccugg aguacaaucu uacuacuacg gagguaguaa 1320
uggagaaugu gacggcuuuu ugggaagagg guuuuggaga acuguuugag aaagcaaagc 1380
agaauaacaa caaccgcaag accucaaaug gggacgauuc ccuguuuuuc ucgaacuucu 1440
cccugcucgg aacacccgug uugaaggaca ucaauuucaa gauugagagg ggacagcuuc 1500
ucgcgguagc gggaagcacu ggugcgggaa aaacuagccu cuugauggug auuauggggg 1560
agcuugagcc cagcgagggg aagauuaaac acuccgggcg uaucucauuc uguagccagu 1620
uuucauggau caugcccgga accauuaaag agaacaucau uuucggagua uccuaugaug 1680
aguaccgaua cagaucgguc auuaaggcgu gccaguugga agaggacauu ucuaaguucg 1740
ccgagaagga uaacaucguc uugggagaag gggguauuac auugucggga gggcagcgag 1800
cgcggaucag ccucgcgaga gcgguauaca aagaugcaga uuuguaucug cuugauucac 1860
cguuuggaua ccucgacgua uugacagaaa aagaaaucuu cgagucgugc guguguaaac 1920
uuauggcuaa uaagacgaga auccugguga caucaaaaau ggaacaccuu aagaaggcgg 1980
acaagauccu gauccuccac gaaggaucgu ccuacuuuua cggcacuuuc ucagaguugc 2040
aaaacuugca gccggacuuc ucaagcaaac ucauggggug ugacucauuc gaccaguuca 2100
gcgcggaacg gcggaacucg aucuugacgg aaacgcugca ccgauucucg cuugagggug 2160
augccccggu aucguggacc gagacaaaga agcagucguu uaagcagaca ggagaauuug 2220
gugagaaaag aaagaacagu aucuugaauc cuauuaacuc aauucgcaag uucucaaucg 2280
uccagaaaac uccacugcag augaauggaa uugaagagga uucggacgaa ccccuggagc 2340
gcaggcuuag ccucgugccg gauucagagc aaggggaggc cauucuuccc cggauuucgg 2400
ugauuucaac cggaccuaca cuucaggcga ggcgaaggca auccgugcuc aaccucauga 2460
cgcauucggu aaaccagggg caaaacauuc accgcaaaac gacggccuca acgagaaaag 2520
ugucacuugc accccaggcg aauuugacug aacucgacau cuacagccgu aggcuuucgc 2580
aagaaaccgg acuugagauc agcgaagaaa ucaaugaaga agauuugaaa gaguguuucu 2640
uugaugacau ggaaucaauc ccagcgguga caacguggaa cacauacuug cguuacauca 2700
cggugcacaa guccuugauu uucguccuca ucuggugucu cgugaucuuu cucgcugagg 2760
ucgcagcguc acuugugguc cucuggcugc uugguaauac gcccuugcaa gacaaaggca 2820
auucuacaca cucaagaaac aauuccuaug ccgugauuau cacuucuaca agcucguauu 2880
acguguuuua caucuacgua ggaguggccg acacucugcu cgcgaugggu uucuuccgag 2940
gacucccacu cguucacacg cuuaucacug ucuccaagau ucuccaccau aagaugcuuc 3000
auagcguacu gcaggcuccc auguccaccu ugaauacgcu caaggcggga gguauuuuga 3060
aucgcuucuc aaaagauauu gcaauuuugg augaccuucu gccccugacg aucuucgacu 3120
ucauccaguu guugcugauc gugauugggg cuauugcagu agucgcuguc cuccagccuu 3180
acauuuuugu cgcgaccguu ccggugaucg uggcguuuau caugcugcgg gccuauuucu 3240
ugcagacguc acagcagcuu aagcaacugg agucugaagg gaggucgccu aucuuuacgc 3300
aucuugugac caguuugaag ggauugugga cguugcgcgc cuuuggcagg cagcccuacu 3360
uugaaacacu guuccacaaa gcgcugaauc uccauacggc aaauugguuu uuguauuuga 3420
guacccuccg augguuucag augcgcauug agaugauuuu ugugaucuuc uuuaucgcgg 3480
ugacuuuuau cuccaucuug accacgggag agggcgaggg acgggucggu auuauccuga 3540
cacucgccau gaacauuaug agcacuuugc agugggcagu gaacagcucg auugaugugg 3600
auagccugau gagguccguu ucgagggucu uuaaguucau cgacaugccg acggagggaa 3660
agcccacaaa aaguacgaaa cccuauaaga augggcaauu gaguaaggua augaucaucg 3720
agaacaguca cgugaagaag gaugacaucu ggccuagcgg gggucagaug accgugaagg 3780
accugacggc aaaauacacc gagggaggga acgcaauccu ugaaaacauc ucguucagca 3840
uuagccccgg ucagcgugug ggguugcucg ggaggaccgg gucaggaaaa ucgacguugc 3900
ugucggccuu cuugagacuu cugaauacag agggugagau ccagaucgac ggcguuucgu 3960
gggauagcau caccuugcag caguggcgga aagcguuugg aguaaucccc caaaaggucu 4020
uuaucuuuag cggaaccuuc cgaaagaauc ucgauccuua ugaacagugg ucagaucaag 4080
agauuuggaa agucgcggac gagguuggcc uucggagugu aaucgagcag uuuccgggaa 4140
aacucgacuu uguccuugua gaugggggau gcguccuguc gcaugggcac aagcagcuca 4200
ugugccuggc gcgauccguc cucucuaaag cgaaaauucu ucucuuggau gaaccuucgg 4260
cccaucugga cccgguaacg uaucagauca ucagaaggac acuuaagcag gcguuugccg 4320
acugcacggu gauucucugu gagcaucgua ucgaggccau gcucgaaugc cagcaauuuc 4380
uugucaucga agagaauaag guccgccagu acgacuccau ccagaagcug cuuaaugaga 4440
gaucauuguu ccggcaggcg auuucaccau ccgauagggu gaaacuuuuu ccacacagaa 4500
auucgucgaa gugcaagucc aaaccgcaga ucgcggccuu gaaagaagag acugaagaag 4560
aaguucaaga cacgcgucuu uaaggguggc aucccuguga ccccucccca gugccucucc 4620
uggcccugga aguugccacu ccagugccca ccagccuugu ccuaauaaaa uuaaguugca 4680
ucaaagcu 4688
Claims (88)
1.一种在体内递送核酸的方法,其包含通过肺部递送向需要递送的个体施用包含以下各者的组合物:
核酸;和
囊封所述核酸的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质。
2.一种在体内递送核酸的方法,其包含通过肺部递送向需要递送的个体施用包含以下各者的组合物:
核酸,其中所述核酸编码囊性纤维化跨膜传导调节(CFTR)蛋白;和
囊封所述核酸的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质。
3.根据权利要求1或2所述的方法,其中所述肺部递送包括喷雾。
4.根据权利要求1或2所述的方法,其中所述个体是需要递送的个体。
5.根据权利要求1或2所述的方法,其中所述脂质纳米颗粒具有至少70%的核酸囊封百分比。
6.根据前述权利要求中任一项所述的方法,其中所述三种不同的脂质组分包含辅助脂质和PEG改性脂质中的一种或两种。
7.根据前述权利要求中任一项所述的方法,其中所述基于固醇的阳离子脂质具有根据式(A)的结构,
B-L1-S (式A),
或其质子化形式,其中
B是碱性官能团,其中所述质子化形式具有不超过约8.0的pKa;
L1是任选经取代的连接基团,其为C1-C20亚烷基或2至20元亚杂烷基;且
S是固醇。
8.根据权利要求7所述的方法,其中B是任选经取代的5元或6元含氮杂芳基。
9.根据权利要求8所述的方法,其中B是选自吡咯基、咪唑基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
10.根据权利要求9所述的方法,其中B是选自吡咯基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
11.根据权利要求10所述的方法,其中B是选自吡咯基、吡唑基、三唑基、四唑基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
12.根据权利要求7至11中任一项所述的方法,其中L1是任选经取代的连接基团,其为C1-C20亚烷基。
13.根据权利要求7至11中任一项所述的方法,其中L1是任选经取代的连接基团,其为2至20元亚杂烷基。
14.根据权利要求13所述的方法,其中L1是非肽的2至20元亚杂烷基。
15.根据权利要求13或14所述的方法,其中L1包含为酯基、酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。
16.根据权利要求15所述的方法,其中L1包含为酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。
17.根据权利要求16所述的方法,其中L1包含为酰胺基、碳酸酯基或脲基的部分。
18.根据权利要求15所述的方法,其中L1为-X1-C(X3)-X2、-(C1-C19亚烷基)-X1-C(X3)-X2、-X1-C(X3)-X2(C1-C19亚烷基)-、-(C1-C19亚烷基)-X1-、-X1-(C1-C19亚烷基)-,
其中
每个X1和X2独立地为共价键、-O-、-S-或-NH-;
X3独立地为=O、=S或=NH;且
其中所述C1-C19亚烷基任选经取代。
19.根据权利要求7至18中任一项所述的方法,其中L1不包含具有结构-N(R')2的取代基,或其带正电形式,其中每个R'独立地为氢或任选经取代的C1-C20烷基。
20.根据权利要求7至19中任一项所述的方法,其中S是动物固醇,或其氧化或还原形式。
21.根据权利要求7至19中任一项所述的方法,其中S是植物固醇,或其氧化或还原形式。
22.根据权利要求7至19中任一项所述的方法,其中S是合成固醇,或其氧化或还原形式。
23.根据权利要求7至19中任一项所述的方法,其中S是选自胆固醇、胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。
24.根据权利要求23所述的方法,其中S是选自胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。
25.根据权利要求24所述的方法,其中S是选自以下的固醇:
其中R是任选经取代的C1-C20烷基。
26.根据权利要求24所述的方法,其中S是选自以下的固醇:
其中R是任选经取代的C1-C20烷基。
27.根据前述权利要求中任一项所述的方法,其中所述基于固醇的阳离子脂质包含咪唑胆固醇酯(ICE)。
28.根据权利要求1至26中任一项所述的方法,其中所述基于固醇的阳离子脂质不包含咪唑胆固醇酯(ICE)。
29.根据前述权利要求中任一项所述的方法,其中所述核酸选自DNA、siRNA、微RNA和/或mRNA。
30.根据前述权利要求中任一项所述的方法,其中所述核酸是编码蛋白质或肽的mRNA。
31.根据权利要求30所述的方法,其中所述编码蛋白质或肽的mRNA是密码子优化的。
32.根据权利要求30所述的方法,其中所述mRNA包含一种或多种经修饰的核苷酸。
33.根据权利要求30所述的方法,其中所述mRNA包含对所述mRNA的5'非翻译区的修饰。
34.根据权利要求33所述的方法,其中所述5'非翻译区的所述修饰包含将Cap1结构包括在内。
35.根据权利要求30所述的方法,其中所述mRNA包含对所述mRNA的3'非翻译区的修饰。
36.根据权利要求35所述的方法,其中所述3'非翻译区的所述修饰包含将poly A尾包括在内。
37.根据前述权利要求中任一项所述的方法,其中所述脂质纳米颗粒具有小于约100nm、95nm、90nm、85nm、80nm、75nm、70nm、65nm、60nm、55nm、50nm、45nm或40nm的尺寸。
38.一种配制用于喷雾的组合物,其包含:
核酸;和
囊封所述核酸的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质,且另外,其中所述脂质纳米颗粒具有至少70%的核酸囊封百分比。
39.一种组合物,其包含:
核酸;和
囊封所述核酸的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质,且
另外,其中所述脂质纳米颗粒具有至少70%的核酸囊封百分比。
40.根据权利要求38或39所述的组合物,其中所述脂质纳米颗粒具有至少85%的核酸囊封百分比。
41.根据权利要求38或39所述的组合物,其中所述脂质纳米颗粒具有至少90%的核酸囊封百分比。
42.一种组合物,其包含:
编码囊性纤维化跨膜传导调节(CFTR)蛋白的mRNA;和
囊封所述mRNA的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质,且
另外,其中所述脂质纳米颗粒具有至少80%的mRNA囊封百分比。
43.根据权利要求42所述的组合物,其中所述脂质纳米颗粒具有至少85%或至少90%的mRNA囊封百分比。
44.一种组合物,其包含:
核酸;和
囊封所述核酸的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质,且
另外,其中所述基于固醇的阳离子脂质占所述总脂质的不超过70%。
45.一种组合物,其包含:
编码囊性纤维化跨膜传导调节(CFTR)蛋白的mRNA;和
囊封所述mRNA的脂质纳米颗粒,其中每个单独的脂质纳米颗粒包含不超过三种不同的脂质组分,一种不同的脂质组分是基于固醇的阳离子脂质,且
另外,其中所述基于固醇的阳离子脂质占所述总脂质的不超过70%。
46.根据权利要求44或45所述的组合物,其中所述基于固醇的阳离子脂质占所述总脂质的不超过65%。
47.根据权利要求44或45所述的组合物,其中所述基于固醇的阳离子脂质占所述总脂质的不超过60%。
48.根据权利要求38至47中任一项所述的组合物,其中所述三种不同的脂质组分包含辅助脂质和PEG改性脂质中的一种或两种。
49.根据权利要求38至48中任一项所述的组合物,其中所述基于固醇的阳离子脂质具有根据式(A)的结构,
B-L1-S (式A),
或其质子化形式,其中
B是碱性官能团,其中所述质子化形式具有不超过约8.0的pKa;
L1是任选经取代的连接基团,其为C1-C20亚烷基或2至20元亚杂烷基;且
S是固醇。
50.根据权利要求49所述的组合物,其中B是任选经取代的5元或6元含氮杂芳基。
51.根据权利要求50所述的组合物,其中B是选自吡咯基、咪唑基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
52.根据权利要求51所述的组合物,其中B是选自吡咯基、吡唑基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
53.根据权利要求52所述的组合物,其中B是选自吡咯基、吡唑基、三唑基、四唑基、嘧啶基、吡嗪基和哒嗪基的基团,其中的每一个任选经取代。
54.根据权利要求49至53中任一项所述的组合物,其中L1是任选经取代的连接基团,其为C1-C20亚烷基。
55.根据权利要求49至53中任一项所述的组合物,其中L1是任选经取代的连接基团,其为2至20元亚杂烷基。
56.根据权利要求55所述的组合物,其中L1是非肽的2至20元亚杂烷基。
57.根据权利要求55或56所述的组合物,其中L1包含为酯基、酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。
58.根据权利要求57所述的组合物,其中L1包含为酰胺基、氨基甲酸酯基、碳酸酯基或脲基的部分。
59.根据权利要求58所述的组合物,其中L1包含为酰胺基、碳酸酯基或脲基的部分。
60.根据权利要求57所述的组合物,其中L1为-X1-C(X3)-X2、-(C1-C19亚烷基)-X1-C(X3)-X2、-X1-C(X3)-X2(C1-C19亚烷基)-、-(C1-C19亚烷基)-X1-、-X1-(C1-C19亚烷基)-,
其中
每个X1和X2独立地为共价键、-O-、-S-或-NH-;
X3独立地为=O、=S或=NH;且
其中所述C1-C19亚烷基任选经取代。
61.根据权利要求49至60中任一项所述的组合物,其中L1不包含具有结构-N(R')2的取代基,或其带正电形式,其中每个R'独立地为氢或任选经取代的C1-C20烷基。
62.根据权利要求49至61中任一项所述的组合物,其中S是动物固醇,或其氧化或还原形式。
63.根据权利要求49至61中任一项所述的组合物,其中S是植物固醇,或其氧化或还原形式。
64.根据权利要求49至61中任一项所述的组合物,其中S是合成固醇,或其氧化或还原形式。
65.根据权利要求49至61中任一项所述的组合物,其中S是选自胆固醇、胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。
66.根据权利要求65所述的组合物,其中S是选自胆固醇的氧化形式、胆固醇的还原形式、烷基石胆酸盐、豆固醇、豆固烷醇、菜油固醇、麦角固醇和谷固醇的固醇。
67.根据权利要求66所述的组合物,其中S是选自以下的固醇:
其中R是任选经取代的C1-C20烷基。
68.根据权利要求66所述的组合物,其中S是选自以下的固醇:
其中R是任选经取代的C1-C20烷基。
69.根据权利要求38至68中任一项所述的组合物,其中所述基于固醇的阳离子脂质包含咪唑胆固醇酯(ICE)。
70.根据权利要求38至68中任一项所述的组合物,其中所述基于固醇的阳离子脂质不包含咪唑胆固醇酯(ICE)。
71.根据权利要求38至41、44或46至70中任一项所述的组合物,其中所述核酸选自DNA、siRNA、微RNA和/或mRNA。
72.根据权利要求38至41或42至70中任一项所述的组合物,其中所述核酸是编码蛋白质或肽的mRNA。
73.根据权利要求42、43或72中任一项所述的组合物,其中所述编码蛋白质或肽的mRNA是密码子优化的。
74.根据权利要求42、43或72中任一项所述的组合物,其中所述mRNA包含一种或多种经修饰的核苷酸。
75.根据权利要求42、43或72中任一项所述的组合物,其中所述mRNA包含对所述mRNA的5'非翻译区的修饰。
76.根据权利要求75所述的组合物,其中所述5’非翻译区的所述修饰包含将Cap1结构包括在内。
77.根据权利要求42、43或72中任一项所述的组合物,其中所述mRNA包含对所述mRNA的3'非翻译区的修饰。
78.根据权利要求77所述的组合物,其中所述3'非翻译区的所述修饰包含将poly A尾包括在内。
79.根据权利要求38至78中任一项所述的组合物,其中所述脂质纳米颗粒具有小于约100nm、95nm、90nm、85nm、80nm、75nm、70nm、65nm、60nm、55nm、50nm、45nm或40nm的尺寸。
80.根据权利要求42、43或45至79中任一项所述的组合物,其中所述编码CFTR蛋白的mRNA包含SEQ ID NO:1。
81.根据权利要求42、43或45至80中任一项所述的组合物,其中所述编码CFTR蛋白的mRNA包含与SEQ ID NO:1至少70%、75%、80%、85%、90%或95%一致的多核苷酸序列。
82.根据权利要求38至81中任一项所述的组合物,其中所述脂质纳米颗粒通过将mRNA溶液和脂质溶液混合到20%乙醇中而形成。
83.根据权利要求38至82中任一项所述的组合物,其中所述脂质纳米颗粒通过切向流过滤进一步纯化。
84.一种在体内递送mRNA的方法,其包含向个体施用根据权利要求42、43或45至83中任一项所述的组合物。
85.根据权利要求84所述的方法,其中所述个体是需要递送的个体。
86.根据权利要求84或85所述的方法,其中所述组合物是静脉内施用的。
87.根据权利要求84或85所述的方法,其中所述组合物通过肺部递送施用。
88.根据权利要求87所述的方法,其中所述肺部递送包含喷雾。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310516878.XA CN116549671A (zh) | 2016-11-10 | 2017-11-10 | 用于递送mrna的改进的基于ice的脂质纳米颗粒制剂 |
Applications Claiming Priority (13)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201662420428P | 2016-11-10 | 2016-11-10 | |
US201662420421P | 2016-11-10 | 2016-11-10 | |
US62/420,421 | 2016-11-10 | ||
US62/420,428 | 2016-11-10 | ||
US201662421021P | 2016-11-11 | 2016-11-11 | |
US201662421007P | 2016-11-11 | 2016-11-11 | |
US62/421,021 | 2016-11-11 | ||
US62/421,007 | 2016-11-11 | ||
US201762464330P | 2017-02-27 | 2017-02-27 | |
US201762464327P | 2017-02-27 | 2017-02-27 | |
US62/464,330 | 2017-02-27 | ||
US62/464,327 | 2017-02-27 | ||
PCT/US2017/061100 WO2018089790A1 (en) | 2016-11-10 | 2017-11-10 | Improved ice-based lipid nanoparticle formulation for delivery of mrna |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310516878.XA Division CN116549671A (zh) | 2016-11-10 | 2017-11-10 | 用于递送mrna的改进的基于ice的脂质纳米颗粒制剂 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN110114058A true CN110114058A (zh) | 2019-08-09 |
CN110114058B CN110114058B (zh) | 2023-05-26 |
Family
ID=62065903
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310516878.XA Pending CN116549671A (zh) | 2016-11-10 | 2017-11-10 | 用于递送mrna的改进的基于ice的脂质纳米颗粒制剂 |
CN201780080493.6A Active CN110114058B (zh) | 2016-11-10 | 2017-11-10 | 用于递送mrna的改进的基于ice的脂质纳米颗粒制剂 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310516878.XA Pending CN116549671A (zh) | 2016-11-10 | 2017-11-10 | 用于递送mrna的改进的基于ice的脂质纳米颗粒制剂 |
Country Status (10)
Country | Link |
---|---|
US (4) | US10471153B2 (zh) |
EP (1) | EP3538068A1 (zh) |
KR (1) | KR20190120160A (zh) |
CN (2) | CN116549671A (zh) |
AU (2) | AU2017357748B2 (zh) |
CA (1) | CA3043033A1 (zh) |
IL (2) | IL266501B1 (zh) |
MA (1) | MA46756A (zh) |
MX (1) | MX2019005470A (zh) |
WO (1) | WO2018089790A1 (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113637708A (zh) * | 2021-08-09 | 2021-11-12 | 中国科学院过程工程研究所 | 一种CRISPR-cas9基因编辑系统递送载体及其制备方法和应用 |
CN115487306A (zh) * | 2022-11-18 | 2022-12-20 | 深圳市华元生物技术股份有限公司 | 一种药物递送载体及其制备方法、应用、糖尿病治疗药物 |
Families Citing this family (68)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
RS57314B1 (sr) | 2009-12-01 | 2018-08-31 | Translate Bio Inc | Isporuka irnk za povećanje ekspresije proteina i enzima u humanim genetskim oboljenjima |
PL2590626T3 (pl) | 2010-07-06 | 2016-04-29 | Glaxosmithkline Biologicals Sa | Liposomy z lipidami o korzystnej wartości pka do dostarczania rna |
JP5940064B2 (ja) | 2010-07-06 | 2016-06-29 | ノバルティス アーゲー | 低用量のrnaを用いた大型哺乳動物の免疫化 |
SI3243526T1 (sl) | 2010-07-06 | 2020-02-28 | Glaxosmithkline Biologicals S.A. | Dostava RNA za sprožitev večih imunskih poti |
HRP20230185T8 (hr) | 2010-08-31 | 2023-07-21 | Glaxosmithkline Biologicals Sa | Pegilirani liposomi za isporuku rna koja kodira imunogen |
EP4098325A1 (en) | 2010-10-11 | 2022-12-07 | GlaxoSmithKline Biologicals S.A. | Antigen delivery platforms |
EP4074693A1 (en) | 2011-06-08 | 2022-10-19 | Translate Bio, Inc. | Cleavable lipids |
WO2013006838A1 (en) | 2011-07-06 | 2013-01-10 | Novartis Ag | Immunogenic combination compositions and uses thereof |
EP3283125B1 (en) | 2015-04-17 | 2021-12-29 | CureVac Real Estate GmbH | Lyophilization of rna |
SG10201910431RA (en) | 2015-05-20 | 2020-01-30 | Curevac Ag | Dry powder composition comprising long-chain rna |
CN107530448A (zh) | 2015-05-20 | 2018-01-02 | 库瑞瓦格股份公司 | 包含长链rna的干粉组合物 |
MA45053A (fr) | 2016-05-18 | 2019-03-27 | Modernatx Inc | Polynucléotides codant pour un régulateur de conductance transmembranaire de fibrose kystique pour le traitement de la fibrose kystique |
WO2018089790A1 (en) | 2016-11-10 | 2018-05-17 | Translate Bio, Inc. | Improved ice-based lipid nanoparticle formulation for delivery of mrna |
CA3054062A1 (en) * | 2017-02-27 | 2018-08-30 | Translate Bio, Inc. | Novel codon-optimized cftr mrna |
WO2018213476A1 (en) * | 2017-05-16 | 2018-11-22 | Translate Bio, Inc. | Treatment of cystic fibrosis by delivery of codon-optimized mrna encoding cftr |
WO2018232357A1 (en) | 2017-06-15 | 2018-12-20 | Modernatx, Inc. | Rna formulations |
JP7275111B2 (ja) | 2017-08-31 | 2023-05-17 | モデルナティエックス インコーポレイテッド | 脂質ナノ粒子の生成方法 |
CN108676867B (zh) * | 2018-06-06 | 2020-06-09 | 青岛泱深生物医药有限公司 | 诊治子痫前期的vwce基因及其应用 |
BR112020025268A2 (pt) * | 2018-06-28 | 2021-03-09 | The University Of North Carolina At Chapel Hill | Cassetes de expressão e genes cln5 otimizados e seu uso |
US11357726B2 (en) | 2018-08-29 | 2022-06-14 | Translate Bio, Inc. | Process of preparing mRNA-loaded lipid nanoparticles |
KR20210068093A (ko) | 2018-09-28 | 2021-06-08 | 넛크래커 테라퓨틱스 인코포레이티드 | 핵산 전달을 위한 지질화 양이온성 펩타이드 화합물을 포함하는 지질 나노입자 제형 |
AU2019359299B2 (en) | 2018-10-09 | 2022-04-21 | The University Of British Columbia | Compositions and systems comprising transfection-competent vesicles free of organic-solvents and detergents and methods related thereto |
US20210378962A1 (en) | 2018-10-19 | 2021-12-09 | Translate Bio, Inc. | Pumpless encapsulation of messenger rna |
AU2019376660A1 (en) | 2018-11-09 | 2021-06-03 | Translate Bio, Inc. | 2,5-dioxopiperazine lipids with intercalated ester, thioester, disulfide and anhydride moieities |
WO2020097511A2 (en) | 2018-11-09 | 2020-05-14 | Translate Bio, Inc. | Messenger rna therapy for treatment of ocular diseases |
CA3120647A1 (en) | 2018-11-21 | 2020-05-28 | Translate Bio, Inc. | Treatment of cystic fibrosis by delivery of nebulized mrna encoding cftr |
CA3125588A1 (en) | 2019-01-07 | 2020-07-16 | Translate Bio, Inc. | Composition and methods for treatment of primary ciliary dyskinesia |
EP3996683A1 (en) | 2019-07-08 | 2022-05-18 | Translate Bio, Inc. | Improved mrna-loaded lipid nanoparticles and processes of making the same |
JP2022542562A (ja) | 2019-07-23 | 2022-10-05 | トランスレイト バイオ, インコーポレイテッド | mRNA担持脂質ナノ粒子の安定性組成物および作製のプロセス |
WO2021021988A1 (en) * | 2019-07-30 | 2021-02-04 | Translate Bio, Inc. | Treatment of cystic fibrosis by delivery of nebulized mrna encoding cftr |
CN110674130A (zh) | 2019-08-30 | 2020-01-10 | 深圳鸿智云创科技有限公司 | 数据传输方法 |
EP4076393A2 (en) | 2019-12-20 | 2022-10-26 | Translate Bio, Inc. | Rectal delivery of messenger rna |
WO2021127641A1 (en) | 2019-12-20 | 2021-06-24 | Translate Bio, Inc. | Improved process of preparing mrna-loaded lipid nanoparticles |
US20230062603A1 (en) * | 2020-01-07 | 2023-03-02 | Translate Bio Ma, Inc. | Formulations for delivery of oligonucleotides to lung cells |
WO2021142245A1 (en) | 2020-01-10 | 2021-07-15 | Translate Bio, Inc. | Compounds, pharmaceutical compositions and methods for modulating expression of muc5b in lung cells and tissues |
US20210324369A1 (en) | 2020-02-10 | 2021-10-21 | Translate Bio, Inc. | Methods and compositions for messenger rna purification |
CA3171495A1 (en) | 2020-02-18 | 2021-08-26 | Translate Bio, Inc. | Improved processes for in vitro transcription of messenger rna |
CN111239416B (zh) * | 2020-02-18 | 2020-11-20 | 无锡市妇幼保健院 | 一种与妊娠期肝内胆汁淤积症辅助诊断相关的血清/血浆蛋白质分子标志物及其应用 |
AU2021226578A1 (en) | 2020-02-25 | 2022-10-20 | Translate Bio, Inc. | Improved processes of preparing mRNA-loaded lipid nanoparticles |
WO2021226463A1 (en) | 2020-05-07 | 2021-11-11 | Translate Bio, Inc. | Composition and methods for treatment of primary ciliary dyskinesia |
IL297962A (en) | 2020-05-07 | 2023-01-01 | Translate Bio Inc | Optimized nucleotide sequences encoding sars-cov-2 antigens |
US20210353556A1 (en) | 2020-05-15 | 2021-11-18 | Translate Bio, Inc. | Lipid Nanoparticle Formulations for mRNA Delivery |
CN112190691B (zh) * | 2020-09-23 | 2022-02-18 | 南方医科大学 | Lifr蛋白作为非酒精性脂肪肝生物标记物及治疗靶点的应用 |
AU2021358777A1 (en) | 2020-10-06 | 2023-06-15 | Translate Bio, Inc. | Improved process and formulation of lipid nanoparticles |
WO2022081548A1 (en) | 2020-10-12 | 2022-04-21 | Translate Bio, Inc. | Improved process of preparing ice-based lipid nanoparticles |
CA3198411A1 (en) | 2020-10-12 | 2022-04-21 | Translate Bio, Inc. | Improved process of preparing mrna-loaded lipid nanoparticles |
TW202233232A (zh) | 2020-11-06 | 2022-09-01 | 法商賽諾菲公司 | 遞送mRNA疫苗的脂質奈米顆粒 |
WO2022099194A1 (en) * | 2020-11-09 | 2022-05-12 | Translate Bio, Inc. | Improved compositions for delivery of codon-optimized mrna |
KR102285906B1 (ko) * | 2021-02-18 | 2021-08-04 | 주식회사 엔에이피 | 수생태계 교란어종 관리를 위한 큰입배스 불임 유도용 조성물 |
EP4313115A1 (en) | 2021-03-25 | 2024-02-07 | Translate Bio, Inc. | Optimized nucleotide sequences encoding the extracellular domain of human ace2 protein or a portion thereof |
CN117203340A (zh) | 2021-04-16 | 2023-12-08 | 基因泰克公司 | 优化的tlr7配体及其用途 |
WO2022225918A1 (en) | 2021-04-19 | 2022-10-27 | Translate Bio, Inc. | Improved compositions for delivery of mrna |
CN115300609A (zh) * | 2021-05-08 | 2022-11-08 | 西湖大学 | 缺氧诱导脂滴相关蛋白(hilpda)在预防或治疗高血脂相关疾病中的应用 |
EP4355308A1 (en) | 2021-06-18 | 2024-04-24 | Sanofi | Multivalent influenza vaccines |
EP4362920A1 (en) | 2021-07-01 | 2024-05-08 | Translate Bio, Inc. | Compositions for delivery of mrna |
CA3230031A1 (en) | 2021-09-03 | 2023-03-09 | Patrick Baumhof | Novel lipid nanoparticles for delivery of nucleic acids |
WO2023073228A1 (en) | 2021-10-29 | 2023-05-04 | CureVac SE | Improved circular rna for expressing therapeutic proteins |
US20230302112A1 (en) | 2021-11-05 | 2023-09-28 | Sanofi | Respiratory synctial virus rna vaccine |
WO2023086893A1 (en) | 2021-11-10 | 2023-05-19 | Translate Bio, Inc. | Composition and methods for treatment of primary ciliary dyskinesia |
US20230310571A1 (en) | 2021-11-30 | 2023-10-05 | Sanofi Pasteur Inc. | Human metapneumovirus vaccines |
WO2023111262A1 (en) | 2021-12-17 | 2023-06-22 | Sanofi | Lyme disease rna vaccine |
WO2023144330A1 (en) | 2022-01-28 | 2023-08-03 | CureVac SE | Nucleic acid encoded transcription factor inhibitors |
WO2023214082A2 (en) | 2022-05-06 | 2023-11-09 | Sanofi | Signal sequences for nucleic acid vaccines |
WO2023227608A1 (en) | 2022-05-25 | 2023-11-30 | Glaxosmithkline Biologicals Sa | Nucleic acid based vaccine encoding an escherichia coli fimh antigenic polypeptide |
US20240043826A1 (en) | 2022-07-29 | 2024-02-08 | Sanofi | Methods for ethanol-free mrna purification |
WO2024028492A1 (en) | 2022-08-04 | 2024-02-08 | Sanofi | Quantitative assessment of rna encapsulation |
WO2024044108A1 (en) | 2022-08-22 | 2024-02-29 | The Henry M. Jackson Foundation For The Advancement Of Military Medicine, Inc. | Vaccines against coronaviruses |
WO2024057237A1 (en) | 2022-09-16 | 2024-03-21 | Pfizer Inc. | Lipid nanoparticles |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0814777A1 (en) * | 1995-01-23 | 1998-01-07 | University Of Pittsburgh | Stable lipid-comprising drug delivery complexes and methods for their production |
US6375980B1 (en) * | 1998-01-08 | 2002-04-23 | Research Development Foundation | Stabilization of lipid:DNA formulations during nebulization |
CN105142676A (zh) * | 2013-03-14 | 2015-12-09 | 夏尔人类遗传性治疗公司 | Cftr mrna组合物以及相关方法和用途 |
US20160151409A1 (en) * | 2013-03-15 | 2016-06-02 | Shire Human Genetic Therapies, Inc. | Synergistic enhancement of the delivery of nucleic acids via blended formulations |
Family Cites Families (24)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5132418A (en) | 1980-02-29 | 1992-07-21 | University Patents, Inc. | Process for preparing polynucleotides |
US4500707A (en) | 1980-02-29 | 1985-02-19 | University Patents, Inc. | Nucleosides useful in the preparation of polynucleotides |
US4458066A (en) | 1980-02-29 | 1984-07-03 | University Patents, Inc. | Process for preparing polynucleotides |
US4415732A (en) | 1981-03-27 | 1983-11-15 | University Patents, Inc. | Phosphoramidite compounds and processes |
US4668777A (en) | 1981-03-27 | 1987-05-26 | University Patents, Inc. | Phosphoramidite nucleoside compounds |
US4973679A (en) | 1981-03-27 | 1990-11-27 | University Patents, Inc. | Process for oligonucleo tide synthesis using phosphormidite intermediates |
US4373071A (en) | 1981-04-30 | 1983-02-08 | City Of Hope Research Institute | Solid-phase synthesis of polynucleotides |
US4401796A (en) | 1981-04-30 | 1983-08-30 | City Of Hope Research Institute | Solid-phase synthesis of polynucleotides |
US4737323A (en) | 1986-02-13 | 1988-04-12 | Liposome Technology, Inc. | Liposome extrusion method |
US5153319A (en) | 1986-03-31 | 1992-10-06 | University Patents, Inc. | Process for preparing polynucleotides |
US5047524A (en) | 1988-12-21 | 1991-09-10 | Applied Biosystems, Inc. | Automated system for polynucleotide synthesis and purification |
US5262530A (en) | 1988-12-21 | 1993-11-16 | Applied Biosystems, Inc. | Automated system for polynucleotide synthesis and purification |
US5885613A (en) | 1994-09-30 | 1999-03-23 | The University Of British Columbia | Bilayer stabilizing components and their use in forming programmable fusogenic liposomes |
US5700642A (en) | 1995-05-22 | 1997-12-23 | Sri International | Oligonucleotide sizing using immobilized cleavable primers |
US5744335A (en) | 1995-09-19 | 1998-04-28 | Mirus Corporation | Process of transfecting a cell with a polynucleotide mixed with an amphipathic compound and a DNA-binding protein |
LT2578685T (lt) | 2005-08-23 | 2019-06-10 | The Trustees Of The University Of Pennsylvania | Rnr, apimančios modifikuotus nukleozidus ir jų panaudojimo būdai |
WO2009030254A1 (en) | 2007-09-04 | 2009-03-12 | Curevac Gmbh | Complexes of rna and cationic peptides for transfection and for immunostimulation |
AU2010278309B2 (en) | 2009-07-31 | 2013-10-31 | Ethris Gmbh | RNA with a combination of unmodified and modified nucleotides for protein expression |
RS57314B1 (sr) * | 2009-12-01 | 2018-08-31 | Translate Bio Inc | Isporuka irnk za povećanje ekspresije proteina i enzima u humanim genetskim oboljenjima |
JP6184945B2 (ja) * | 2011-06-08 | 2017-08-23 | シャイアー ヒューマン ジェネティック セラピーズ インコーポレイテッド | mRNA送達のための脂質ナノ粒子組成物および方法 |
EP2828290B1 (en) * | 2012-03-23 | 2018-08-15 | The United States of America, represented by the Secretary, Department of Health and Human Services | Anti-mesothelin chimeric antigen receptors |
EP2971102B8 (en) | 2013-03-14 | 2018-08-15 | Translate Bio, Inc. | Quantitative assessment for cap efficiency of messenger rna |
WO2018089790A1 (en) * | 2016-11-10 | 2018-05-17 | Translate Bio, Inc. | Improved ice-based lipid nanoparticle formulation for delivery of mrna |
WO2019207060A1 (en) | 2018-04-25 | 2019-10-31 | Ethris Gmbh | Lipid-based formulations for the delivery of rna |
-
2017
- 2017-11-10 WO PCT/US2017/061100 patent/WO2018089790A1/en unknown
- 2017-11-10 IL IL266501A patent/IL266501B1/en unknown
- 2017-11-10 CN CN202310516878.XA patent/CN116549671A/zh active Pending
- 2017-11-10 KR KR1020197016564A patent/KR20190120160A/ko not_active IP Right Cessation
- 2017-11-10 MA MA046756A patent/MA46756A/fr unknown
- 2017-11-10 CN CN201780080493.6A patent/CN110114058B/zh active Active
- 2017-11-10 US US15/809,605 patent/US10471153B2/en active Active
- 2017-11-10 MX MX2019005470A patent/MX2019005470A/es unknown
- 2017-11-10 CA CA3043033A patent/CA3043033A1/en active Pending
- 2017-11-10 IL IL310050A patent/IL310050A/en unknown
- 2017-11-10 AU AU2017357748A patent/AU2017357748B2/en active Active
- 2017-11-10 EP EP17804762.7A patent/EP3538068A1/en active Pending
-
2019
- 2019-10-11 US US16/599,928 patent/US11013812B2/en active Active
-
2020
- 2020-01-27 US US16/773,638 patent/US10940207B2/en active Active
-
2021
- 2021-04-23 US US17/239,131 patent/US20210353761A1/en active Pending
-
2024
- 2024-02-07 AU AU2024200738A patent/AU2024200738A1/en not_active Abandoned
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0814777A1 (en) * | 1995-01-23 | 1998-01-07 | University Of Pittsburgh | Stable lipid-comprising drug delivery complexes and methods for their production |
US6375980B1 (en) * | 1998-01-08 | 2002-04-23 | Research Development Foundation | Stabilization of lipid:DNA formulations during nebulization |
CN105142676A (zh) * | 2013-03-14 | 2015-12-09 | 夏尔人类遗传性治疗公司 | Cftr mrna组合物以及相关方法和用途 |
US20160151409A1 (en) * | 2013-03-15 | 2016-06-02 | Shire Human Genetic Therapies, Inc. | Synergistic enhancement of the delivery of nucleic acids via blended formulations |
Non-Patent Citations (2)
Title |
---|
RUIZ F E等: "A Clinical Inflammatory Syndrome Attributable to Aerosolized Lipid–DNA Administration in Cystic Fibrosis", 《HUMAN GENE THERAPY》 * |
RUIZ F E等: "A Clinical Inflammatory Syndrome Attributable to Aerosolized Lipid–DNA Administration in Cystic Fibrosis", 《HUMAN GENE THERAPY》, vol. 12, no. 7, 6 July 2004 (2004-07-06) * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113637708A (zh) * | 2021-08-09 | 2021-11-12 | 中国科学院过程工程研究所 | 一种CRISPR-cas9基因编辑系统递送载体及其制备方法和应用 |
CN115487306A (zh) * | 2022-11-18 | 2022-12-20 | 深圳市华元生物技术股份有限公司 | 一种药物递送载体及其制备方法、应用、糖尿病治疗药物 |
Also Published As
Publication number | Publication date |
---|---|
IL310050A (en) | 2024-03-01 |
US11013812B2 (en) | 2021-05-25 |
EP3538068A1 (en) | 2019-09-18 |
MA46756A (fr) | 2019-09-18 |
US20200155691A1 (en) | 2020-05-21 |
US10471153B2 (en) | 2019-11-12 |
WO2018089790A1 (en) | 2018-05-17 |
US10940207B2 (en) | 2021-03-09 |
CA3043033A1 (en) | 2018-05-17 |
CN110114058B (zh) | 2023-05-26 |
KR20190120160A (ko) | 2019-10-23 |
MX2019005470A (es) | 2019-11-21 |
IL266501A (en) | 2019-07-31 |
IL266501B1 (en) | 2024-02-01 |
US20180125989A1 (en) | 2018-05-10 |
US20210353761A1 (en) | 2021-11-18 |
AU2017357748B2 (en) | 2023-11-09 |
AU2024200738A1 (en) | 2024-02-29 |
AU2017357748A1 (en) | 2019-06-06 |
CN116549671A (zh) | 2023-08-08 |
US20200038515A1 (en) | 2020-02-06 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN110114058A (zh) | 用于递送mrna的改进的基于ice的脂质纳米颗粒制剂 | |
JP6913699B2 (ja) | 非肺標的細胞へのmRNAの経肺送達 | |
EP3060257B1 (en) | Lipid formulations for delivery of messenger rna | |
JP2023041951A (ja) | mRNAのCNS送達及びその使用方法 | |
CN106413811A (zh) | 精氨基琥珀酸合成酶缺乏症的mrna疗法 | |
JP6506749B2 (ja) | フェニルケトン尿症のためのmRNA療法 | |
CN109072223A (zh) | 多聚体编码核酸及其用途 | |
JP2019533707A (ja) | Mrna担持脂質ナノ粒子を調製する改善されたプロセス | |
AU2021232818B2 (en) | Mrna therapy for pompe disease | |
CN109312313A (zh) | 用于治疗鸟氨酸转氨甲酰酶缺乏症的信使rna疗法 | |
US20220193247A1 (en) | Novel Codon-Optimized CFTR MRNA | |
CN104334161A (zh) | 可电离的阳离子脂质 | |
JP2022532213A (ja) | Mrna担持脂質ナノ粒子を調製する改善されたプロセス | |
JP2000516209A (ja) | カチオン性両親媒性物質/dna複合体 | |
CN104487055A (zh) | 脂质衍生的中性纳米颗粒 | |
CN106795142A (zh) | 用于递送核酸的立体化学富集组合物 | |
JP2023524071A (ja) | 嚢胞性線維症を治療するための核酸及び方法 | |
JP2024505449A (ja) | 生分解性特徴を含むナノ材料 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |