CN115612674A - 一种溶瘤病毒及其用途 - Google Patents
一种溶瘤病毒及其用途 Download PDFInfo
- Publication number
- CN115612674A CN115612674A CN202110798531.XA CN202110798531A CN115612674A CN 115612674 A CN115612674 A CN 115612674A CN 202110798531 A CN202110798531 A CN 202110798531A CN 115612674 A CN115612674 A CN 115612674A
- Authority
- CN
- China
- Prior art keywords
- ala
- gly
- leu
- thr
- pro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 244000309459 oncolytic virus Species 0.000 title claims abstract description 264
- 108091006027 G proteins Proteins 0.000 claims abstract description 153
- 108091000058 GTP-Binding Proteins 0.000 claims abstract description 153
- 102000030782 GTP binding Human genes 0.000 claims abstract description 151
- 238000006467 substitution reaction Methods 0.000 claims abstract description 109
- 101710085938 Matrix protein Proteins 0.000 claims abstract description 46
- 101710127721 Membrane protein Proteins 0.000 claims abstract description 46
- 238000000034 method Methods 0.000 claims abstract description 30
- 241000700605 Viruses Species 0.000 claims abstract description 29
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 24
- 239000013604 expression vector Substances 0.000 claims abstract description 21
- 239000003814 drug Substances 0.000 claims abstract description 9
- 210000004027 cell Anatomy 0.000 claims description 219
- 235000001014 amino acid Nutrition 0.000 claims description 145
- 150000001413 amino acids Chemical group 0.000 claims description 81
- 230000035772 mutation Effects 0.000 claims description 64
- 150000007523 nucleic acids Chemical class 0.000 claims description 40
- 238000002360 preparation method Methods 0.000 claims description 38
- 206010028980 Neoplasm Diseases 0.000 claims description 33
- 108090000623 proteins and genes Proteins 0.000 claims description 30
- 230000002147 killing effect Effects 0.000 claims description 29
- 241000711975 Vesicular stomatitis virus Species 0.000 claims description 27
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims description 20
- 102000039446 nucleic acids Human genes 0.000 claims description 20
- 108020004707 nucleic acids Proteins 0.000 claims description 20
- 239000004474 valine Substances 0.000 claims description 20
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 18
- 125000002987 valine group Chemical group [H]N([H])C([H])(C(*)=O)C([H])(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 16
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 14
- 235000018102 proteins Nutrition 0.000 claims description 14
- 102000004169 proteins and genes Human genes 0.000 claims description 14
- 210000004881 tumor cell Anatomy 0.000 claims description 14
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 13
- 235000003704 aspartic acid Nutrition 0.000 claims description 12
- CKLJMWTZIZZHCS-REOHCLBHSA-N aspartic acid group Chemical group N[C@@H](CC(=O)O)C(=O)O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 12
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 12
- 201000010099 disease Diseases 0.000 claims description 10
- 239000004475 Arginine Substances 0.000 claims description 9
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 9
- 101001065501 Escherichia phage MS2 Lysis protein Proteins 0.000 claims description 8
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 8
- 239000004471 Glycine Substances 0.000 claims description 8
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims description 8
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 8
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims description 8
- 239000004473 Threonine Substances 0.000 claims description 8
- 235000004279 alanine Nutrition 0.000 claims description 8
- 235000013922 glutamic acid Nutrition 0.000 claims description 8
- 239000004220 glutamic acid Substances 0.000 claims description 8
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims description 8
- 229960000310 isoleucine Drugs 0.000 claims description 8
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims description 8
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 8
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 claims description 7
- 101710141454 Nucleoprotein Proteins 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 6
- 125000000341 threoninyl group Chemical group [H]OC([H])(C([H])([H])[H])C([H])(N([H])[H])C(*)=O 0.000 claims description 6
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 5
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 5
- 101710181008 P protein Proteins 0.000 claims description 5
- 101710177166 Phosphoprotein Proteins 0.000 claims description 5
- 239000003937 drug carrier Substances 0.000 claims description 5
- 229930182817 methionine Natural products 0.000 claims description 5
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 4
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims description 4
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 4
- 239000004472 Lysine Substances 0.000 claims description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 4
- 102100034574 P protein Human genes 0.000 claims description 4
- 201000011510 cancer Diseases 0.000 claims description 4
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 230000002265 prevention Effects 0.000 claims description 4
- 230000002062 proliferating effect Effects 0.000 claims description 4
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 claims description 4
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 claims description 3
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 claims description 3
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 claims description 3
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 claims description 3
- 125000001493 tyrosinyl group Chemical group [H]OC1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 claims description 3
- 201000005787 hematologic cancer Diseases 0.000 claims description 2
- 208000037819 metastatic cancer Diseases 0.000 claims description 2
- 208000011575 metastatic malignant neoplasm Diseases 0.000 claims description 2
- 230000002459 sustained effect Effects 0.000 claims description 2
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 claims 1
- 125000003275 alpha amino acid group Chemical group 0.000 abstract description 116
- 229940079593 drug Drugs 0.000 abstract description 5
- 108010050848 glycylleucine Proteins 0.000 description 104
- 108010061238 threonyl-glycine Proteins 0.000 description 96
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 84
- UQJNXZSSGQIPIQ-FBCQKBJTSA-N Gly-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)CN UQJNXZSSGQIPIQ-FBCQKBJTSA-N 0.000 description 83
- 108010047495 alanylglycine Proteins 0.000 description 80
- WQLDNOCHHRISMS-NAKRPEOUSA-N Ala-Pro-Ile Chemical compound [H]N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WQLDNOCHHRISMS-NAKRPEOUSA-N 0.000 description 52
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 50
- RTZCUEHYUQZIDE-WHFBIAKZSA-N Ala-Ser-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RTZCUEHYUQZIDE-WHFBIAKZSA-N 0.000 description 48
- IOFVWPYSRSCWHI-JXUBOQSCSA-N Ala-Thr-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](C)N IOFVWPYSRSCWHI-JXUBOQSCSA-N 0.000 description 48
- JRDYDYXZKFNNRQ-XPUUQOCRSA-N Gly-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN JRDYDYXZKFNNRQ-XPUUQOCRSA-N 0.000 description 48
- WSGXUIQTEZDVHJ-GARJFASQSA-N Leu-Ala-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(O)=O WSGXUIQTEZDVHJ-GARJFASQSA-N 0.000 description 48
- XBBKIIGCUMBKCO-JXUBOQSCSA-N Leu-Ala-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XBBKIIGCUMBKCO-JXUBOQSCSA-N 0.000 description 48
- DPURXCQCHSQPAN-AVGNSLFASA-N Leu-Pro-Pro Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DPURXCQCHSQPAN-AVGNSLFASA-N 0.000 description 48
- SBVPYBFMIGDIDX-SRVKXCTJSA-N Pro-Pro-Pro Chemical compound OC(=O)[C@@H]1CCCN1C(=O)[C@H]1N(C(=O)[C@H]2NCCC2)CCC1 SBVPYBFMIGDIDX-SRVKXCTJSA-N 0.000 description 48
- 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 48
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 48
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 48
- 229940024606 amino acid Drugs 0.000 description 41
- PYTZFYUXZZHOAD-WHFBIAKZSA-N Gly-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)CN PYTZFYUXZZHOAD-WHFBIAKZSA-N 0.000 description 36
- XPJBQTCXPJNIFE-ZETCQYMHSA-N Gly-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)CN XPJBQTCXPJNIFE-ZETCQYMHSA-N 0.000 description 36
- JQFILXICXLDTRR-FBCQKBJTSA-N Gly-Thr-Gly Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)NCC(O)=O JQFILXICXLDTRR-FBCQKBJTSA-N 0.000 description 36
- 108010027668 glycyl-alanyl-valine Proteins 0.000 description 36
- QSDKBRMVXSWAQE-BFHQHQDPSA-N Gly-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN QSDKBRMVXSWAQE-BFHQHQDPSA-N 0.000 description 30
- 108010040030 histidinoalanine Proteins 0.000 description 29
- SOBIAADAMRHGKH-CIUDSAMLSA-N Ala-Leu-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O SOBIAADAMRHGKH-CIUDSAMLSA-N 0.000 description 28
- 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 28
- SXWQMBGNFXAGAT-FJXKBIBVSA-N Met-Gly-Thr Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O SXWQMBGNFXAGAT-FJXKBIBVSA-N 0.000 description 28
- AFFKUNVPPLQUGA-DCAQKATOSA-N Met-Leu-Ala Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O AFFKUNVPPLQUGA-DCAQKATOSA-N 0.000 description 28
- 108010079364 N-glycylalanine Proteins 0.000 description 28
- BRKHVZNDAOMAHX-BIIVOSGPSA-N Ser-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N BRKHVZNDAOMAHX-BIIVOSGPSA-N 0.000 description 28
- MROIJTGJGIDEEJ-RCWTZXSCSA-N Thr-Pro-Pro Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 MROIJTGJGIDEEJ-RCWTZXSCSA-N 0.000 description 28
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 28
- KUFVXLQLDHJVOG-SHGPDSBTSA-N Ala-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](C)N)O KUFVXLQLDHJVOG-SHGPDSBTSA-N 0.000 description 27
- 239000013612 plasmid Substances 0.000 description 26
- OBVSBEYOMDWLRJ-BFHQHQDPSA-N Ala-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N OBVSBEYOMDWLRJ-BFHQHQDPSA-N 0.000 description 25
- AAWLEICNDUHIJM-MBLNEYKQSA-N Ala-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C)N)O AAWLEICNDUHIJM-MBLNEYKQSA-N 0.000 description 25
- VWWKKDNCCLAGRM-GVXVVHGQSA-N (2s)-2-[[2-[[(2s)-2-[[(2s)-2-amino-4-methylpentanoyl]amino]propanoyl]amino]acetyl]amino]-3-methylbutanoic acid Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O VWWKKDNCCLAGRM-GVXVVHGQSA-N 0.000 description 24
- RLMISHABBKUNFO-WHFBIAKZSA-N Ala-Ala-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O RLMISHABBKUNFO-WHFBIAKZSA-N 0.000 description 24
- CXZFXHGJJPVUJE-CIUDSAMLSA-N Ala-Cys-Leu Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(=O)O)N CXZFXHGJJPVUJE-CIUDSAMLSA-N 0.000 description 24
- HOVPGJUNRLMIOZ-CIUDSAMLSA-N Ala-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C)N HOVPGJUNRLMIOZ-CIUDSAMLSA-N 0.000 description 24
- HIIJOGIBQXHFKE-HHKYUTTNSA-N Ala-Thr-Ala-Pro Chemical compound C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O HIIJOGIBQXHFKE-HHKYUTTNSA-N 0.000 description 24
- KTXKIYXZQFWJKB-VZFHVOOUSA-N Ala-Thr-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O KTXKIYXZQFWJKB-VZFHVOOUSA-N 0.000 description 24
- PKNIZMPLMSKROD-BIIVOSGPSA-N Cys-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CS)N PKNIZMPLMSKROD-BIIVOSGPSA-N 0.000 description 24
- PRXCTTWKGJAPMT-ZLUOBGJFSA-N Cys-Ala-Ser Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O PRXCTTWKGJAPMT-ZLUOBGJFSA-N 0.000 description 24
- YMUFWNJHVPQNQD-ZKWXMUAHSA-N Gly-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN YMUFWNJHVPQNQD-ZKWXMUAHSA-N 0.000 description 24
- IDOGEHIWMJMAHT-BYPYZUCNSA-N Gly-Gly-Cys Chemical compound NCC(=O)NCC(=O)N[C@@H](CS)C(O)=O IDOGEHIWMJMAHT-BYPYZUCNSA-N 0.000 description 24
- ADZGCWWDPFDHCY-ZETCQYMHSA-N Gly-His-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 ADZGCWWDPFDHCY-ZETCQYMHSA-N 0.000 description 24
- YIFUFYZELCMPJP-YUMQZZPRSA-N Gly-Leu-Cys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(O)=O YIFUFYZELCMPJP-YUMQZZPRSA-N 0.000 description 24
- CCBIBMKQNXHNIN-ZETCQYMHSA-N Gly-Leu-Gly Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O CCBIBMKQNXHNIN-ZETCQYMHSA-N 0.000 description 24
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 24
- GGLIDLCEPDHEJO-BQBZGAKWSA-N Gly-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)CN GGLIDLCEPDHEJO-BQBZGAKWSA-N 0.000 description 24
- HFPVRZWORNJRRC-UWVGGRQHSA-N Gly-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN HFPVRZWORNJRRC-UWVGGRQHSA-N 0.000 description 24
- TVTZEOHWHUVYCG-KYNKHSRBSA-N Gly-Thr-Thr Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O TVTZEOHWHUVYCG-KYNKHSRBSA-N 0.000 description 24
- CUVBTVWFVIIDOC-YEPSODPASA-N Gly-Thr-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)CN CUVBTVWFVIIDOC-YEPSODPASA-N 0.000 description 24
- GWCJMBNBFYBQCV-XPUUQOCRSA-N Gly-Val-Ala Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O GWCJMBNBFYBQCV-XPUUQOCRSA-N 0.000 description 24
- BAYQNCWLXIDLHX-ONGXEEELSA-N Gly-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN BAYQNCWLXIDLHX-ONGXEEELSA-N 0.000 description 24
- AFMOTCMSEBITOE-YEPSODPASA-N Gly-Val-Thr Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O AFMOTCMSEBITOE-YEPSODPASA-N 0.000 description 24
- ORZGPQXISSXQGW-IHRRRGAJSA-N His-His-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C(C)C)C(O)=O ORZGPQXISSXQGW-IHRRRGAJSA-N 0.000 description 24
- VYUXYMRNGALHEA-DLOVCJGASA-N His-Leu-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O VYUXYMRNGALHEA-DLOVCJGASA-N 0.000 description 24
- LVXFNTIIGOQBMD-SRVKXCTJSA-N His-Leu-Ser Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O LVXFNTIIGOQBMD-SRVKXCTJSA-N 0.000 description 24
- HDOYNXLPTRQLAD-JBDRJPRFSA-N Ile-Ala-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)O)N HDOYNXLPTRQLAD-JBDRJPRFSA-N 0.000 description 24
- WEWCEPOYKANMGZ-MMWGEVLESA-N Ile-Cys-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)N1CCC[C@@H]1C(=O)O)N WEWCEPOYKANMGZ-MMWGEVLESA-N 0.000 description 24
- NZOCIWKZUVUNDW-ZKWXMUAHSA-N Ile-Gly-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O NZOCIWKZUVUNDW-ZKWXMUAHSA-N 0.000 description 24
- KXUKTDGKLAOCQK-LSJOCFKGSA-N Ile-Val-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O KXUKTDGKLAOCQK-LSJOCFKGSA-N 0.000 description 24
- RQZFWBLDTBDEOF-RNJOBUHISA-N Ile-Val-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N RQZFWBLDTBDEOF-RNJOBUHISA-N 0.000 description 24
- UGTHTQWIQKEDEH-BQBZGAKWSA-N L-alanyl-L-prolylglycine zwitterion Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UGTHTQWIQKEDEH-BQBZGAKWSA-N 0.000 description 24
- VWHGTYCRDRBSFI-ZETCQYMHSA-N Leu-Gly-Gly Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)NCC(O)=O VWHGTYCRDRBSFI-ZETCQYMHSA-N 0.000 description 24
- VGPCJSXPPOQPBK-YUMQZZPRSA-N Leu-Gly-Ser Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O VGPCJSXPPOQPBK-YUMQZZPRSA-N 0.000 description 24
- 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 24
- SEMUSFOBZGKBGW-YTFOTSKYSA-N Leu-Ile-Ile 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)CC)C(O)=O SEMUSFOBZGKBGW-YTFOTSKYSA-N 0.000 description 24
- HRTRLSRYZZKPCO-BJDJZHNGSA-N Leu-Ile-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HRTRLSRYZZKPCO-BJDJZHNGSA-N 0.000 description 24
- WMIOEVKKYIMVKI-DCAQKATOSA-N Leu-Pro-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O WMIOEVKKYIMVKI-DCAQKATOSA-N 0.000 description 24
- YUTNOGOMBNYPFH-XUXIUFHCSA-N Leu-Pro-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YUTNOGOMBNYPFH-XUXIUFHCSA-N 0.000 description 24
- 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 24
- CHJKEDSZNSONPS-DCAQKATOSA-N Leu-Pro-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O CHJKEDSZNSONPS-DCAQKATOSA-N 0.000 description 24
- PWPBLZXWFXJFHE-RHYQMDGZSA-N Leu-Pro-Thr Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O PWPBLZXWFXJFHE-RHYQMDGZSA-N 0.000 description 24
- IRMLZWSRWSGTOP-CIUDSAMLSA-N Leu-Ser-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O IRMLZWSRWSGTOP-CIUDSAMLSA-N 0.000 description 24
- ZJZNLRVCZWUONM-JXUBOQSCSA-N Leu-Thr-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O ZJZNLRVCZWUONM-JXUBOQSCSA-N 0.000 description 24
- QGQGAIBGTUJRBR-NAKRPEOUSA-N Met-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCSC QGQGAIBGTUJRBR-NAKRPEOUSA-N 0.000 description 24
- OFNCSQNBSWGGNV-DCAQKATOSA-N Met-Cys-His Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 OFNCSQNBSWGGNV-DCAQKATOSA-N 0.000 description 24
- FWAHLGXNBLWIKB-NAKRPEOUSA-N Met-Ile-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCSC FWAHLGXNBLWIKB-NAKRPEOUSA-N 0.000 description 24
- CQRGINSEMFBACV-WPRPVWTQSA-N Met-Val-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O CQRGINSEMFBACV-WPRPVWTQSA-N 0.000 description 24
- 108010065395 Neuropep-1 Proteins 0.000 description 24
- WSRWHZRUOCACLJ-UWVGGRQHSA-N Pro-Gly-His Chemical compound C([C@@H](C(=O)O)NC(=O)CNC(=O)[C@H]1NCCC1)C1=CN=CN1 WSRWHZRUOCACLJ-UWVGGRQHSA-N 0.000 description 24
- FEVDNIBDCRKMER-IUCAKERBSA-N Pro-Gly-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)CNC(=O)[C@@H]1CCCN1 FEVDNIBDCRKMER-IUCAKERBSA-N 0.000 description 24
- PEYNRYREGPAOAK-LSJOCFKGSA-N Pro-His-Ala Chemical compound C([C@@H](C(=O)N[C@@H](C)C([O-])=O)NC(=O)[C@H]1[NH2+]CCC1)C1=CN=CN1 PEYNRYREGPAOAK-LSJOCFKGSA-N 0.000 description 24
- BFXZQMWKTYWGCF-PYJNHQTQSA-N Pro-His-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BFXZQMWKTYWGCF-PYJNHQTQSA-N 0.000 description 24
- IBGCFJDLCYTKPW-NAKRPEOUSA-N Pro-Ile-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]1CCCN1 IBGCFJDLCYTKPW-NAKRPEOUSA-N 0.000 description 24
- MKGIILKDUGDRRO-FXQIFTODSA-N Pro-Ser-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H]1CCCN1 MKGIILKDUGDRRO-FXQIFTODSA-N 0.000 description 24
- PRKWBYCXBBSLSK-GUBZILKMSA-N Pro-Ser-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O PRKWBYCXBBSLSK-GUBZILKMSA-N 0.000 description 24
- HRNQLKCLPVKZNE-CIUDSAMLSA-N Ser-Ala-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O HRNQLKCLPVKZNE-CIUDSAMLSA-N 0.000 description 24
- RNFKSBPHLTZHLU-WHFBIAKZSA-N Ser-Cys-Gly Chemical compound C([C@@H](C(=O)N[C@@H](CS)C(=O)NCC(=O)O)N)O RNFKSBPHLTZHLU-WHFBIAKZSA-N 0.000 description 24
- UQFYNFTYDHUIMI-WHFBIAKZSA-N Ser-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CO UQFYNFTYDHUIMI-WHFBIAKZSA-N 0.000 description 24
- SFTZTYBXIXLRGQ-JBDRJPRFSA-N Ser-Ile-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O SFTZTYBXIXLRGQ-JBDRJPRFSA-N 0.000 description 24
- IAORETPTUDBBGV-CIUDSAMLSA-N Ser-Leu-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CO)N IAORETPTUDBBGV-CIUDSAMLSA-N 0.000 description 24
- HEUVHBXOVZONPU-BJDJZHNGSA-N Ser-Leu-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HEUVHBXOVZONPU-BJDJZHNGSA-N 0.000 description 24
- FLONGDPORFIVQW-XGEHTFHBSA-N Ser-Pro-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO FLONGDPORFIVQW-XGEHTFHBSA-N 0.000 description 24
- CKDXFSPMIDSMGV-GUBZILKMSA-N Ser-Pro-Val Chemical compound [H]N[C@@H](CO)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O CKDXFSPMIDSMGV-GUBZILKMSA-N 0.000 description 24
- JCLAFVNDBJMLBC-JBDRJPRFSA-N Ser-Ser-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JCLAFVNDBJMLBC-JBDRJPRFSA-N 0.000 description 24
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 24
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 24
- VLMIUSLQONKLDV-HEIBUPTGSA-N Ser-Thr-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VLMIUSLQONKLDV-HEIBUPTGSA-N 0.000 description 24
- PMTWIUBUQRGCSB-FXQIFTODSA-N Ser-Val-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O PMTWIUBUQRGCSB-FXQIFTODSA-N 0.000 description 24
- CAJFZCICSVBOJK-SHGPDSBTSA-N Thr-Ala-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CAJFZCICSVBOJK-SHGPDSBTSA-N 0.000 description 24
- KWQBJOUOSNJDRR-XAVMHZPKSA-N Thr-Cys-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)N1CCC[C@@H]1C(=O)O)N)O KWQBJOUOSNJDRR-XAVMHZPKSA-N 0.000 description 24
- SLUWOCTZVGMURC-BFHQHQDPSA-N Thr-Gly-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O SLUWOCTZVGMURC-BFHQHQDPSA-N 0.000 description 24
- KBBRNEDOYWMIJP-KYNKHSRBSA-N Thr-Gly-Thr Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O KBBRNEDOYWMIJP-KYNKHSRBSA-N 0.000 description 24
- KRGDDWVBBDLPSJ-CUJWVEQBSA-N Thr-His-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(O)=O KRGDDWVBBDLPSJ-CUJWVEQBSA-N 0.000 description 24
- GMXIJHCBTZDAPD-QPHKQPEJSA-N Thr-Ile-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N GMXIJHCBTZDAPD-QPHKQPEJSA-N 0.000 description 24
- GXUWHVZYDAHFSV-FLBSBUHZSA-N Thr-Ile-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GXUWHVZYDAHFSV-FLBSBUHZSA-N 0.000 description 24
- YOOAQCZYZHGUAZ-KATARQTJSA-N Thr-Leu-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YOOAQCZYZHGUAZ-KATARQTJSA-N 0.000 description 24
- QFEYTTHKPSOFLV-OSUNSFLBSA-N Thr-Met-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H]([C@@H](C)O)N QFEYTTHKPSOFLV-OSUNSFLBSA-N 0.000 description 24
- WTMPKZWHRCMMMT-KZVJFYERSA-N Thr-Pro-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O WTMPKZWHRCMMMT-KZVJFYERSA-N 0.000 description 24
- UQCNIMDPYICBTR-KYNKHSRBSA-N Thr-Thr-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O UQCNIMDPYICBTR-KYNKHSRBSA-N 0.000 description 24
- BBPCSGKKPJUYRB-UVOCVTCTSA-N Thr-Thr-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O BBPCSGKKPJUYRB-UVOCVTCTSA-N 0.000 description 24
- 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 24
- 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 24
- FEFZWCSXEMVSPO-LSJOCFKGSA-N Val-His-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](C)C(O)=O FEFZWCSXEMVSPO-LSJOCFKGSA-N 0.000 description 24
- UMPVMAYCLYMYGA-ONGXEEELSA-N Val-Leu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O UMPVMAYCLYMYGA-ONGXEEELSA-N 0.000 description 24
- SSYBNWFXCFNRFN-GUBZILKMSA-N Val-Pro-Ser Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O SSYBNWFXCFNRFN-GUBZILKMSA-N 0.000 description 24
- 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 24
- 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 24
- DVLWZWNAQUBZBC-ZNSHCXBVSA-N Val-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N)O DVLWZWNAQUBZBC-ZNSHCXBVSA-N 0.000 description 24
- JAIZPWVHPQRYOU-ZJDVBMNYSA-N Val-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](C(C)C)N)O JAIZPWVHPQRYOU-ZJDVBMNYSA-N 0.000 description 24
- 108010016616 cysteinylglycine Proteins 0.000 description 24
- 108010090037 glycyl-alanyl-isoleucine Proteins 0.000 description 24
- 108010084264 glycyl-glycyl-cysteine Proteins 0.000 description 24
- 108010026364 glycyl-glycyl-leucine Proteins 0.000 description 24
- 108010033719 glycyl-histidyl-glycine Proteins 0.000 description 24
- 108010085325 histidylproline Proteins 0.000 description 24
- 108010009932 leucyl-alanyl-glycyl-valine Proteins 0.000 description 24
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 24
- 108010029020 prolylglycine Proteins 0.000 description 24
- 108010026333 seryl-proline Proteins 0.000 description 24
- 108010071207 serylmethionine Proteins 0.000 description 24
- 108010015385 valyl-prolyl-proline Proteins 0.000 description 24
- 238000000338 in vitro Methods 0.000 description 17
- 208000015181 infectious disease Diseases 0.000 description 17
- XQLBWXHVZVBNJM-FXQIFTODSA-N Pro-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 XQLBWXHVZVBNJM-FXQIFTODSA-N 0.000 description 16
- URPSJRMWHQTARR-MBLNEYKQSA-N Thr-Ile-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O URPSJRMWHQTARR-MBLNEYKQSA-N 0.000 description 16
- NBTGEURICRTMGL-WHFBIAKZSA-N Ala-Gly-Ser Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O NBTGEURICRTMGL-WHFBIAKZSA-N 0.000 description 15
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 15
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 12
- OTEWWRBKGONZBW-UHFFFAOYSA-N 2-[[2-[[2-[(2-azaniumylacetyl)amino]-4-methylpentanoyl]amino]acetyl]amino]acetate Chemical compound NCC(=O)NC(CC(C)C)C(=O)NCC(=O)NCC(O)=O OTEWWRBKGONZBW-UHFFFAOYSA-N 0.000 description 12
- DCVYRWFAMZFSDA-ZLUOBGJFSA-N Ala-Ser-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O DCVYRWFAMZFSDA-ZLUOBGJFSA-N 0.000 description 12
- UGVQELHRNUDMAA-BYPYZUCNSA-N Gly-Ala-Gly Chemical compound [NH3+]CC(=O)N[C@@H](C)C(=O)NCC([O-])=O UGVQELHRNUDMAA-BYPYZUCNSA-N 0.000 description 12
- VSVZIEVNUYDAFR-YUMQZZPRSA-N Gly-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN VSVZIEVNUYDAFR-YUMQZZPRSA-N 0.000 description 12
- UHPAZODVFFYEEL-QWRGUYRKSA-N Gly-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)CN UHPAZODVFFYEEL-QWRGUYRKSA-N 0.000 description 12
- MIIVFRCYJABHTQ-ONGXEEELSA-N Gly-Leu-Val Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O MIIVFRCYJABHTQ-ONGXEEELSA-N 0.000 description 12
- BMWFDYIYBAFROD-WPRPVWTQSA-N Gly-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN BMWFDYIYBAFROD-WPRPVWTQSA-N 0.000 description 12
- YNMQUIVKEFRCPH-QSFUFRPTSA-N Ile-Ile-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)O)N YNMQUIVKEFRCPH-QSFUFRPTSA-N 0.000 description 12
- CGBYDGAJHSOGFQ-LPEHRKFASA-N Pro-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2 CGBYDGAJHSOGFQ-LPEHRKFASA-N 0.000 description 12
- ZMLRZBWCXPQADC-TUAOUCFPSA-N Pro-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2 ZMLRZBWCXPQADC-TUAOUCFPSA-N 0.000 description 12
- ZUGXSSFMTXKHJS-ZLUOBGJFSA-N Ser-Ala-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O ZUGXSSFMTXKHJS-ZLUOBGJFSA-N 0.000 description 12
- JPIDMRXXNMIVKY-VZFHVOOUSA-N Ser-Ala-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JPIDMRXXNMIVKY-VZFHVOOUSA-N 0.000 description 12
- MSIYNSBKKVMGFO-BHNWBGBOSA-N Thr-Gly-Pro Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N1CCC[C@@H]1C(=O)O)N)O MSIYNSBKKVMGFO-BHNWBGBOSA-N 0.000 description 12
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 12
- 108010020432 prolyl-prolylisoleucine Proteins 0.000 description 12
- 238000001890 transfection Methods 0.000 description 12
- 238000012258 culturing Methods 0.000 description 11
- 238000001514 detection method Methods 0.000 description 11
- 238000003752 polymerase chain reaction Methods 0.000 description 9
- 238000002741 site-directed mutagenesis Methods 0.000 description 9
- PAWIVEIWWYGBAM-YUMQZZPRSA-N Gly-Leu-Ala Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O PAWIVEIWWYGBAM-YUMQZZPRSA-N 0.000 description 8
- CNMOKANDJMLAIF-CIQUZCHMSA-N Ile-Thr-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O CNMOKANDJMLAIF-CIQUZCHMSA-N 0.000 description 8
- RCYUBVHMVUHEBM-RCWTZXSCSA-N Pro-Pro-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O RCYUBVHMVUHEBM-RCWTZXSCSA-N 0.000 description 8
- 108010070944 alanylhistidine Proteins 0.000 description 8
- 238000012163 sequencing technique Methods 0.000 description 8
- 230000007613 environmental effect Effects 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 230000006378 damage Effects 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000011259 mixed solution Substances 0.000 description 6
- 108090000467 Interferon-beta Proteins 0.000 description 5
- 239000012124 Opti-MEM Substances 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000000825 pharmaceutical preparation Substances 0.000 description 5
- 229940127557 pharmaceutical product Drugs 0.000 description 5
- 210000003501 vero cell Anatomy 0.000 description 5
- 230000003612 virological effect Effects 0.000 description 5
- ZFXQNADNEBRERM-BJDJZHNGSA-N Ala-Ala-Pro-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 ZFXQNADNEBRERM-BJDJZHNGSA-N 0.000 description 4
- JBVSSSZFNTXJDX-YTLHQDLWSA-N Ala-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N JBVSSSZFNTXJDX-YTLHQDLWSA-N 0.000 description 4
- PCIFXPRIFWKWLK-YUMQZZPRSA-N Ala-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N PCIFXPRIFWKWLK-YUMQZZPRSA-N 0.000 description 4
- IPZQNYYAYVRKKK-FXQIFTODSA-N Ala-Pro-Ala Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O IPZQNYYAYVRKKK-FXQIFTODSA-N 0.000 description 4
- ADSGHMXEAZJJNF-DCAQKATOSA-N Ala-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C)N ADSGHMXEAZJJNF-DCAQKATOSA-N 0.000 description 4
- BHTBAVZSZCQZPT-GUBZILKMSA-N Ala-Pro-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C)N BHTBAVZSZCQZPT-GUBZILKMSA-N 0.000 description 4
- OLVCTPPSXNRGKV-GUBZILKMSA-N Ala-Pro-Pro Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 OLVCTPPSXNRGKV-GUBZILKMSA-N 0.000 description 4
- XWFWAXPOLRTDFZ-FXQIFTODSA-N Ala-Pro-Ser Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O XWFWAXPOLRTDFZ-FXQIFTODSA-N 0.000 description 4
- REWSWYIDQIELBE-FXQIFTODSA-N Ala-Val-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O REWSWYIDQIELBE-FXQIFTODSA-N 0.000 description 4
- DZLQXIFVQFTFJY-BYPYZUCNSA-N Cys-Gly-Gly Chemical compound SC[C@H](N)C(=O)NCC(=O)NCC(O)=O DZLQXIFVQFTFJY-BYPYZUCNSA-N 0.000 description 4
- SWQALSGKVLYKDT-UHFFFAOYSA-N Gly-Ile-Ala Natural products NCC(=O)NC(C(C)CC)C(=O)NC(C)C(O)=O SWQALSGKVLYKDT-UHFFFAOYSA-N 0.000 description 4
- DBJYVKDPGIFXFO-BQBZGAKWSA-N Gly-Met-Ala Chemical compound [H]NCC(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O DBJYVKDPGIFXFO-BQBZGAKWSA-N 0.000 description 4
- IPIVXQQRZXEUGW-UWJYBYFXSA-N His-Ala-His Chemical compound C([C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 IPIVXQQRZXEUGW-UWJYBYFXSA-N 0.000 description 4
- WSEITRHJRVDTRX-QTKMDUPCSA-N His-Met-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CC1=CN=CN1)N)O WSEITRHJRVDTRX-QTKMDUPCSA-N 0.000 description 4
- ZVKDCQVQTGYBQT-LSJOCFKGSA-N His-Pro-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O ZVKDCQVQTGYBQT-LSJOCFKGSA-N 0.000 description 4
- BRQKGRLDDDQWQJ-MBLNEYKQSA-N His-Thr-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O BRQKGRLDDDQWQJ-MBLNEYKQSA-N 0.000 description 4
- 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 4
- 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 4
- CCQLQKZTXZBXTN-NHCYSSNCSA-N Leu-Gly-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O CCQLQKZTXZBXTN-NHCYSSNCSA-N 0.000 description 4
- KUIDCYNIEJBZBU-AJNGGQMLSA-N Leu-Ile-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O KUIDCYNIEJBZBU-AJNGGQMLSA-N 0.000 description 4
- GNRPTBRHRRZCMA-RWMBFGLXSA-N Leu-Met-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N1CCC[C@@H]1C(=O)O)N GNRPTBRHRRZCMA-RWMBFGLXSA-N 0.000 description 4
- 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 4
- MUCIDQMDOYQYBR-IHRRRGAJSA-N Leu-Pro-His Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N MUCIDQMDOYQYBR-IHRRRGAJSA-N 0.000 description 4
- URHJPNHRQMQGOZ-RHYQMDGZSA-N Leu-Thr-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(O)=O URHJPNHRQMQGOZ-RHYQMDGZSA-N 0.000 description 4
- MYAPQOBHGWJZOM-UWVGGRQHSA-N Met-Gly-Leu Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(C)C MYAPQOBHGWJZOM-UWVGGRQHSA-N 0.000 description 4
- 108010061100 Nucleoproteins Proteins 0.000 description 4
- AFXCXDQNRXTSBD-FJXKBIBVSA-N Pro-Gly-Thr Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O AFXCXDQNRXTSBD-FJXKBIBVSA-N 0.000 description 4
- KDBHVPXBQADZKY-GUBZILKMSA-N Pro-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 KDBHVPXBQADZKY-GUBZILKMSA-N 0.000 description 4
- DCHQYSOGURGJST-FJXKBIBVSA-N Pro-Thr-Gly Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O DCHQYSOGURGJST-FJXKBIBVSA-N 0.000 description 4
- VMLONWHIORGALA-SRVKXCTJSA-N Ser-Leu-Leu Chemical compound CC(C)C[C@@H](C([O-])=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]([NH3+])CO VMLONWHIORGALA-SRVKXCTJSA-N 0.000 description 4
- KCGIREHVWRXNDH-GARJFASQSA-N Ser-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N KCGIREHVWRXNDH-GARJFASQSA-N 0.000 description 4
- 101710137500 T7 RNA polymerase Proteins 0.000 description 4
- QQWNRERCGGZOKG-WEDXCCLWSA-N Thr-Gly-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O QQWNRERCGGZOKG-WEDXCCLWSA-N 0.000 description 4
- RFKVQLIXNVEOMB-WEDXCCLWSA-N Thr-Leu-Gly Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)O)N)O RFKVQLIXNVEOMB-WEDXCCLWSA-N 0.000 description 4
- MXDOAJQRJBMGMO-FJXKBIBVSA-N Thr-Pro-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O MXDOAJQRJBMGMO-FJXKBIBVSA-N 0.000 description 4
- FWTFAZKJORVTIR-VZFHVOOUSA-N Thr-Ser-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O FWTFAZKJORVTIR-VZFHVOOUSA-N 0.000 description 4
- AZSHAZJLOZQYAY-FXQIFTODSA-N Val-Ala-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O AZSHAZJLOZQYAY-FXQIFTODSA-N 0.000 description 4
- SJRUJQFQVLMZFW-WPRPVWTQSA-N Val-Pro-Gly Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O SJRUJQFQVLMZFW-WPRPVWTQSA-N 0.000 description 4
- 108010087924 alanylproline Proteins 0.000 description 4
- 238000012217 deletion Methods 0.000 description 4
- 230000037430 deletion Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 108010057821 leucylproline Proteins 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 108010077112 prolyl-proline Proteins 0.000 description 4
- 108010093296 prolyl-prolyl-alanine Proteins 0.000 description 4
- 108010090894 prolylleucine Proteins 0.000 description 4
- 108010048818 seryl-histidine Proteins 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 238000000246 agarose gel electrophoresis Methods 0.000 description 3
- 125000000539 amino acid group Chemical group 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 238000010276 construction Methods 0.000 description 3
- 208000035475 disorder Diseases 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000009169 immunotherapy Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000010839 reverse transcription Methods 0.000 description 3
- 238000011895 specific detection Methods 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 239000012096 transfection reagent Substances 0.000 description 3
- 239000013598 vector Substances 0.000 description 3
- OZFAFGSSMRRTDW-UHFFFAOYSA-N (2,4-dichlorophenyl) benzenesulfonate Chemical compound ClC1=CC(Cl)=CC=C1OS(=O)(=O)C1=CC=CC=C1 OZFAFGSSMRRTDW-UHFFFAOYSA-N 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 2
- 108020005544 Antisense RNA Proteins 0.000 description 2
- 101100263837 Bovine ephemeral fever virus (strain BB7721) beta gene Proteins 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 2
- 101100316840 Enterobacteria phage P4 Beta gene Proteins 0.000 description 2
- LRQXRHGQEVWGPV-NHCYSSNCSA-N Gly-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)CN LRQXRHGQEVWGPV-NHCYSSNCSA-N 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- SHVFUCSSACPBTF-VGDYDELISA-N Ile-Ser-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N SHVFUCSSACPBTF-VGDYDELISA-N 0.000 description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 2
- 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 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 102000011931 Nucleoproteins Human genes 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 108010089430 Phosphoproteins Proteins 0.000 description 2
- 102000007982 Phosphoproteins Human genes 0.000 description 2
- FMLRRBDLBJLJIK-DCAQKATOSA-N Pro-Leu-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 FMLRRBDLBJLJIK-DCAQKATOSA-N 0.000 description 2
- 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 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 238000007792 addition Methods 0.000 description 2
- 230000002238 attenuated effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000013066 combination product Substances 0.000 description 2
- 229940127555 combination product Drugs 0.000 description 2
- 239000003184 complementary RNA Substances 0.000 description 2
- 230000001086 cytosolic effect Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 238000001361 intraarterial administration Methods 0.000 description 2
- 238000007918 intramuscular administration Methods 0.000 description 2
- 238000007912 intraperitoneal administration Methods 0.000 description 2
- 238000007913 intrathecal administration Methods 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 108010026228 mRNA guanylyltransferase Proteins 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 238000012858 packaging process Methods 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 239000013600 plasmid vector Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 230000000699 topical effect Effects 0.000 description 2
- CWEAKSWWKHGTRJ-BQBZGAKWSA-N Ala-Gly-Met Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O CWEAKSWWKHGTRJ-BQBZGAKWSA-N 0.000 description 1
- 241000711404 Avian avulavirus 1 Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 241000709687 Coxsackievirus Species 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 241001466953 Echovirus Species 0.000 description 1
- 241000991587 Enterovirus C Species 0.000 description 1
- 101710091045 Envelope protein Proteins 0.000 description 1
- 101710204837 Envelope small membrane protein Proteins 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- YPWHUFAAMNHMGS-QSFUFRPTSA-N Ile-Ala-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N YPWHUFAAMNHMGS-QSFUFRPTSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 101150062031 L gene Proteins 0.000 description 1
- 101710145006 Lysis protein Proteins 0.000 description 1
- 241000712079 Measles morbillivirus Species 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 108090001074 Nucleocapsid Proteins Proteins 0.000 description 1
- 238000010806 PrimeScriptTM RT Reagent kit Methods 0.000 description 1
- 101710188315 Protein X Proteins 0.000 description 1
- 241000702263 Reovirus sp. Species 0.000 description 1
- 108700008625 Reporter Genes Proteins 0.000 description 1
- 241000710961 Semliki Forest virus Species 0.000 description 1
- 241000837158 Senecavirus A Species 0.000 description 1
- 241000700584 Simplexvirus Species 0.000 description 1
- 102100021696 Syncytin-1 Human genes 0.000 description 1
- 108010046722 Thrombospondin 1 Proteins 0.000 description 1
- 102100036034 Thrombospondin-1 Human genes 0.000 description 1
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 1
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- KRAHMIJVUPUOTQ-DCAQKATOSA-N Val-Ser-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N KRAHMIJVUPUOTQ-DCAQKATOSA-N 0.000 description 1
- 108010065667 Viral Matrix Proteins Proteins 0.000 description 1
- 108020000999 Viral RNA Proteins 0.000 description 1
- 239000003070 absorption delaying agent Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- -1 coatings Substances 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000002612 dispersion medium Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 125000000291 glutamic acid group Chemical group N[C@@H](CCC(O)=O)C(=O)* 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 208000019691 hematopoietic and lymphoid cell neoplasm Diseases 0.000 description 1
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 1
- 230000005099 host tropism Effects 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 125000000741 isoleucyl group Chemical group [H]N([H])C(C(C([H])([H])[H])C([H])([H])C([H])([H])[H])C(=O)O* 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 210000004216 mammary stem cell Anatomy 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 230000000174 oncolytic effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000010079 rubber tapping Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 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 1
- 230000005909 tumor killing Effects 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- 210000002845 virion Anatomy 0.000 description 1
- 230000001018 virulence Effects 0.000 description 1
- 230000007923 virulence factor Effects 0.000 description 1
- 239000000304 virulence factor Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/76—Viruses; Subviral particles; Bacteriophages
- A61K35/766—Rhabdovirus, e.g. vesicular stomatitis virus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0684—Cells of the urinary tract or kidneys
- C12N5/0686—Kidney cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2510/00—Genetically modified cells
- C12N2510/02—Cells for production
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/20011—Rhabdoviridae
- C12N2760/20211—Vesiculovirus, e.g. vesicular stomatitis Indiana virus
- C12N2760/20221—Viruses as such, e.g. new isolates, mutants or their genomic sequences
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/20011—Rhabdoviridae
- C12N2760/20211—Vesiculovirus, e.g. vesicular stomatitis Indiana virus
- C12N2760/20222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/20011—Rhabdoviridae
- C12N2760/20211—Vesiculovirus, e.g. vesicular stomatitis Indiana virus
- C12N2760/20232—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/20011—Rhabdoviridae
- C12N2760/20211—Vesiculovirus, e.g. vesicular stomatitis Indiana virus
- C12N2760/20233—Use of viral protein as therapeutic agent other than vaccine, e.g. apoptosis inducing or anti-inflammatory
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/20011—Rhabdoviridae
- C12N2760/20211—Vesiculovirus, e.g. vesicular stomatitis Indiana virus
- C12N2760/20241—Use of virus, viral particle or viral elements as a vector
- C12N2760/20243—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/20011—Rhabdoviridae
- C12N2760/20211—Vesiculovirus, e.g. vesicular stomatitis Indiana virus
- C12N2760/20251—Methods of production or purification of viral material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/20011—Rhabdoviridae
- C12N2760/20211—Vesiculovirus, e.g. vesicular stomatitis Indiana virus
- C12N2760/20251—Methods of production or purification of viral material
- C12N2760/20252—Methods of production or purification of viral material relating to complementing cells and packaging systems for producing virus or viral particles
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2800/00—Nucleic acids vectors
- C12N2800/10—Plasmid DNA
- C12N2800/106—Plasmid DNA for vertebrates
- C12N2800/107—Plasmid DNA for vertebrates for mammalian
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Immunology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Urology & Nephrology (AREA)
- Oncology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Cell Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本申请涉及生物医药的技术领域，具体公开了一种溶瘤病毒及其用途。该溶瘤病毒包括M蛋白和G蛋白；所述M蛋白与SEQ ID NO 1所示的氨基酸序列相比，包含在第51位、第221位和第226位的氨基酸取代；所述G蛋白与SEQ ID NO 2所示的氨基酸序列相比，包含至少一个氨基酸取代；以及该溶瘤病毒的表达载体、生产该溶瘤病毒的病毒生产细胞、包括该溶瘤病毒的药物组合物；以及上述溶瘤病毒、溶瘤病毒表达载体、病毒生产细胞和/或药物组合物的制备方法和用途。本申请提供的溶瘤病毒具有较好的安全性和治愈率。
Description
技术领域
本申请涉及生物医药的技术领域，更具体地说，它涉及一种溶瘤病毒及其用途。
背景技术
溶瘤病毒是一类具有复制能力的肿瘤杀伤性病毒，目前已被大众接受作为肿瘤免疫治疗的重要分支。溶瘤病毒能够特异性地靶向感染肿瘤细胞，例如利用肿瘤细胞中抑癌基因的失活或缺陷，从而选择性地感染肿瘤细胞；溶瘤病毒感染肿瘤细胞后，会在肿瘤细胞内大量复制并最终摧毁肿瘤细胞，从而杀死肿瘤细胞。同时，溶瘤病毒还可以提供提高宿主自身抗癌反应所必需的免疫刺激信号，从而吸引更多的免疫细胞来继续杀死残余的肿瘤细胞。
虽然溶瘤病毒在肿瘤免疫治疗方面具有较好的应用前景，但野生型溶瘤病毒往往会引起机体神经系统炎症等问题，在利用野生型病毒感染肿瘤细胞的过程中，还存在较大的致病风险。因此，为进一步推进溶瘤病毒的临床应用，需要对野生型溶瘤病毒进行改造，以获得减毒后的溶瘤病毒。将减毒后的溶瘤病毒用于临床应用，从而降低溶瘤病毒的致病风险，提高溶瘤病毒的安全性。
然而，在对溶瘤病毒改造的过程中，若仅是对野生型溶瘤病毒随机进行基因修饰，虽然能够降低其毒性，但改造后的溶瘤病毒可能治愈率效果不佳，甚至改造后的溶瘤病毒无法进行包装，不利于推进溶瘤病毒的临床应用。因此，提供一种安全性能良好，同时治愈率高的改造后的溶瘤病毒,对肿瘤免疫治疗领域具有重要的科研价值和应用意义。
发明内容
本申请提供一种溶瘤病毒及其用途，本申请提供的溶瘤病毒具有良好的安全性和治愈率。
第一方面，本申请提供一种溶瘤病毒，采用如下的技术方案：
在本申请中，所述溶瘤病毒包括M蛋白和G蛋白；所述M蛋白与SEQ ID NO 1所示的氨基酸序列相比，包含在第51位、第221位和第226位的氨基酸取代；所述G蛋白与SEQ ID NO2所示的氨基酸序列相比，包含至少一个氨基酸取代。
在某些实施方式中，所述M蛋白的氨基酸取代包含在第51位的甲硫氨酸突变为精氨酸(M51R)；和/或，在第221位的缬氨酸突变为苯丙氨酸(V221F)；和/或，在第226位的丝氨酸突变为精氨酸(S226R)。
在某些实施方式中，所述M蛋白的氨基酸取代包含在第51位的甲硫氨酸突变为精氨酸(M51R)。
在某些实施方式中，所述M蛋白的氨基酸取代包含在第221位的缬氨酸突变为苯丙氨酸(V221F)。
在某些实施方式中，所述M蛋白的氨基酸取代包含在第226位的丝氨酸突变为精氨酸(S226R)。
在某些实施方式中，所述M蛋白具有M51R和V221F的氨基酸取代。
在某些实施方式中，所述M蛋白具有M51R和S226R的氨基酸取代。
在某些实施方式中，所述M蛋白具有M51R、V221F和S226R的氨基酸取代。
在某些实施方式中，所述M蛋白具有V221F和S226R的氨基酸取代。
在某些实施方式中，所述M蛋白包含如SEQ ID NO 5所示的氨基酸序列。
在某些实施方式中，所述M蛋白包含如SEQ ID NO 3所示的氨基酸序列。
在某些实施方式中，所述M蛋白包含如SEQ ID NO 4所示的氨基酸序列。
在某些实施方式中，所述G蛋白与SEQ ID NO 2所示的氨基酸序列相比，包含以下一个或多个位点的氨基酸取代：第438位、第453位、第471位和第487位。
在某些实施方式中，所述G蛋白与SEQ ID NO 2所示的氨基酸序列相比，包含以下一个或多个位点的氨基酸取代：第53位、第141位、第172位、第217位、第232位、第331位、第371位、第436位、第438位、第453位、第471位和第487位。
在某些实施方式中，所述G蛋白的氨基酸取代包含在第53位的缬氨酸突变为异亮氨酸(V53I)；和/或，在第141位的丙氨酸突变为缬氨酸(A141V)；和/或，在第172位的天冬氨酸突变为酪氨酸(D172Y)；和/或，在第217位的赖氨酸突变为谷氨酸(K217E)；和/或，在第232位的天冬氨酸突变为甘氨酸(D232G)；和/或，在第331位的缬氨酸突变为丙氨酸(V331A)；和/或，在第371位的缬氨酸突变为谷氨酸(V371E)；和/或，在第436位的甘氨酸突变为天冬氨酸(G436D)；和/或，在第438位的苏氨酸突变为丝氨酸(T438S)；和/或，在第453位的苯丙氨酸突变为亮氨酸(F453L)；和/或，在第471位的苏氨酸突变为异亮氨酸(T471I)；和/或，在第487位的酪氨酸突变为组氨酸(Y487H)。
在某些实施方式中，所述G蛋白的氨基酸取代包含在第53位的缬氨酸突变为异亮氨酸(V53I)。
在某些实施方式中，所述G蛋白的氨基酸取代包含在第141位的丙氨酸突变为缬氨酸(A141V)；
在某些实施方式中，所述G蛋白的氨基酸取代包含在第172位的天冬氨酸突变为酪氨酸(D172Y)；
在某些实施方式中，所述G蛋白的氨基酸取代包含在第217位的赖氨酸突变为谷氨酸(K217E)
在某些实施方式中，所述G蛋白的氨基酸取代包含在第232位的天冬氨酸突变为甘氨酸(D232G)；
在某些实施方式中，所述G蛋白的氨基酸取代包含在第331位的缬氨酸突变为丙氨酸(V331A)；
在某些实施方式中，所述G蛋白的氨基酸取代包含在第371位的缬氨酸突变为谷氨酸(V371E)；
在某些实施方式中，所述G蛋白的氨基酸取代包含在第436位的甘氨酸突变为天冬氨酸(G436D)；
在某些实施方式中，所述G蛋白的氨基酸取代包含在第438位的苏氨酸突变为丝氨酸(T438S)；
在某些实施方式中，所述G蛋白的氨基酸取代包含在第453位的苯丙氨酸突变为亮氨酸(F453L)；
在某些实施方式中，所述G蛋白的氨基酸取代包含在第471位的苏氨酸突变为异亮氨酸(T471I)；
在某些实施方式中，所述G蛋白的氨基酸取代包含在第487位的酪氨酸突变为组氨酸(Y487H)。
在某些实施方式中，所述G蛋白的氨基酸取代包含T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I和A141V。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V和D172Y。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y和K217E。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E和D232G。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G和V331A。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A和V371E。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E和G436D。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D和T438S。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S和F453L。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L和T471I。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含V331A、V371E、G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含V371E、G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白的氨基酸取代为V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 17所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 6所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 7所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 8所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 9所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 10所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 11所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 12所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 13所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 14所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 15所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 16所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 18所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 19所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 20所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 21所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 22所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 23所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 24所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 25所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 26所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 27所示的氨基酸序列。
在某些实施方式中，所述G蛋白包含如SEQ ID NO 28所示的氨基酸序列。
在某些实施方式中，所述溶瘤病毒是以棒状病毒为基础，进行定点突变后获得的。
在某些实施方式中，所述溶瘤病毒是以Vesicular Stomatitis Virus(简称“VSV”)病毒为基础，进行定点突变后获得的。
在某些实施方式中，所述溶瘤病毒是以VSV病毒Indiana MuddSummer亚型为基础，进行定点突变后获得的。
在某些实施方式中，所述溶瘤病毒包含或表达外源目的蛋白。
在某些实施方式中，所述溶瘤病毒包含核酸分子，所述核酸分子包含编码所述具有氨基酸取代的M蛋白的核酸序列和编码所述具有氨基酸取代的G蛋白的核酸序列。
在某些实施方式中，所述核酸分子包含编码所述外源目的蛋白的核酸序列。
在某些实施方式中，所述核酸分子中所述编码外源目的蛋白的核酸序列位于所述编码所述具有氨基酸取代的M蛋白的核酸序列和所述编码所述具有氨基酸取代的G蛋白的核酸序列之间。
在某些实施方式中，所述核酸分子编码所述溶瘤病毒的N蛋白(核蛋白N)、L蛋白(大聚合酶蛋白L)和P蛋白(磷酸蛋白P)。
第二方面，本申请提供一种溶瘤病毒表达载体，采用如下的技术方案：
一种溶瘤病毒表达载体，所述溶瘤病毒表达载体能够产生本申请所述的任一项溶瘤病毒。
第三方面，本申请提供一种病毒生产细胞，采用如下的技术方案：
一种病毒生产细胞，所述病毒生产细胞能够产生本申请所述的任一项溶瘤病毒。
第四方面，本申请提供一种药物组合物，采用如下的技术方案：
一种药物组合物，所述药物组合物包括本申请所述的任一项溶瘤病毒，以及任选地药学上可接受的载剂。
第五方面，本申请提供上述溶瘤病毒、溶瘤病毒表达载体、病毒生产细胞和/或药物组合物的制备方法。
第六方面，本申请提供上述溶瘤病毒、溶瘤病毒表达载体、病毒生产细胞和/或药物组合物在制备用于预防和/或治疗疾病和/或病症的药物中的用途。
在某些实施方式中，所述溶瘤病毒、溶瘤病毒表达载体、病毒生产细胞和/或药物组合物用于缓慢持续杀伤异常增生性细胞的方法中。
在某些实施方式中，所述疾病和/或病症包括：异常增生性细胞，选自肿瘤细胞或肿瘤组织的相关细胞；优选的，所述肿瘤细胞是癌细胞；更优选的，所述癌细胞是转移性癌细胞。
在某些实施方式中，所述肿瘤包括实体瘤和/或血液瘤。
综上所述，本申请具有以下有益效果：
本申请提供的溶瘤病毒均对LLC细胞、MC38细胞以及Hela细胞具有较好的侵染能力，对LLC细胞、MC38细胞以及4T1细胞均具有较好的体外杀伤能力，在LLC细胞、MC38细胞以及Hela细胞内均不容易清除。并且，本申请提供的溶瘤病毒均对MEF细胞具有较差的侵染能力、体外杀伤能力以及容易清除度。因此，本申请提供的溶瘤病毒能够较好的用于肿瘤、癌症等细胞的侵染和杀伤，且在肿瘤、癌症等细胞内不易清除，进一步提高了溶瘤病毒对肿瘤、癌症等细胞的治愈率；同时，上述提供的溶瘤病毒不会损伤正常细胞，且在正常细胞内时更加容易清除，进一步保证了正常细胞的安全性。
附图说明
本申请所涉及的发明的具体特征如所附权利要求书所显示。通过参考下文中详细描述的示例性实施方式和附图能够更好地理解本申请所涉及发明的特点和优势。对附图简要说明如下：
图1为本申请制备的溶瘤病毒以及野生型溶瘤病毒对LLC细胞的侵染能力的检测结果。
图2为本申请制备的溶瘤病毒以及野生型溶瘤病毒对MC38细胞的侵染能力的检测结果。
图3为本申请制备的溶瘤病毒以及野生型溶瘤病毒对Hela细胞的侵染能力的检测结果。
图4为本申请制备的溶瘤病毒以及野生型溶瘤病毒对MEF细胞的侵染能力的检测结果。
图5为本申请制备的溶瘤病毒以及野生型溶瘤病毒对LLC细胞的体外杀伤能力的检测结果。
图6为本申请制备的溶瘤病毒以及野生型溶瘤病毒对MC38细胞的体外杀伤能力的检测结果。
图7为本申请制备的溶瘤病毒以及野生型溶瘤病毒对4T1细胞的体外杀伤能力的检测结果。
图8为本申请制备的溶瘤病毒以及野生型溶瘤病毒对MEF细胞的体外杀伤能力的检测结果。
图9为本申请制备的溶瘤病毒以及野生型溶瘤病毒在LLC细胞内的清除难易度的检测结果。
图10为本申请制备的溶瘤病毒以及野生型溶瘤病毒在MC38细胞的清除难易度的检测结果。
图11为本申请制备的溶瘤病毒以及野生型溶瘤病毒在Hela细胞的清除难易度的检测结果。
图12为本申请制备的溶瘤病毒以及野生型溶瘤病毒在MEF细胞的清除难易度的检测结果。
以上附图中，横坐标0表示野生型溶瘤病毒；横坐标1-26分别表示制备例1-26制备的溶瘤病毒；
纵坐标Log10 TCID50表示利用Karber法计算得出的TCID50值，Log10 TCID50的值越大，表示溶瘤病毒对该细胞的侵染能力越好；Log10 TCID50的值越小，表示溶瘤病毒对该细胞的侵染能力越差；
纵坐标OD570表示细胞的OD值，OD570的值越大，表示溶瘤病毒对该细胞的杀伤能力越差；OD570的值越小，表示溶瘤病毒对该细胞的杀伤能力越好。
纵坐标IFN-β水平表示IFN-β基因的表达情况，IFN-β水平的值越大，表示溶瘤病毒在该细胞内越容易清除；IFN-β水平的值越小，表示溶瘤病毒在该细胞内越不容易清除。
本领域技术人员能够从下文的详细描述中容易地洞察到本申请的其它方面和优势。下文的详细描述中仅显示和描述了本申请的示例性实施方式。如本领域技术人员将认识到的，本申请的内容使得本领域技术人员能够对所公开的具体实施方式进行改动而不脱离本申请所涉及发明的精神和范围。相应地，本申请的附图和说明书中的描述仅仅是示例性的，而非为限制性的。
具体实施方式
以下由特定的具体实施例说明本申请发明的实施方式，本领域技术人员可由本说明书所公开的内容容易地了解本申请发明的其他优点及效果。
术语定义
在本申请中，术语“溶瘤病毒”通常指能够在肿瘤细胞中复制并杀死肿瘤细胞的病毒。溶瘤病毒包括但不限于：VSV病毒、痘病毒、单纯疱疹病毒、麻疹病毒、塞姆利基森林病毒、脊髓灰质炎病毒、呼肠孤病毒、塞内卡谷病毒、埃可型肠道病毒、柯萨奇病毒、新城疫病毒和马拉巴病毒。在某些实施方案中，所述溶瘤病毒被改造以提高对肿瘤细胞的选择性。在某些实施方案中，所述溶瘤病毒被改造以降低其免疫原性。在一些实施方案中，本申请所述溶瘤病毒为VSV病毒。在一些实施方案中，VSV病毒为VSV病毒Indiana MuddSummer亚型株的突变体。在一些实施方案中，可以对VSV病毒的M蛋白和G蛋白进行定点基因突变。
在某些实施方式中，本申请所述溶瘤病毒可以是经过基因水平改造的溶瘤病毒，如经过一个或多个基因的修饰来改造，从而提高其肿瘤选择性和/或在分裂细胞中优先复制。所述基因水平的改造可以为修饰参与DNA复制、核酸代谢、宿主向性、表面附着、毒力、裂解和扩散过程的基因，也可以为整合外源基因的改造。所述外源基因可以包括外源免疫调节基因、外源筛选基因、外源报告基因等。所述经过改造的溶瘤病毒也可以是经过氨基酸水平改造的溶瘤病毒，如一个或多个氨基酸的插入、缺失、置换。
在本申请中，术语“M蛋白”通常指VSV病毒基质蛋白。M蛋白是VSV病毒重要的毒力因子，也是VSV病毒中已知可干扰小鼠天然免疫应答的蛋白。术语“M蛋白”还包括其同源物、直系同源物、变体、功能活性片段等。在本申请中，野生型VSV病毒Indiana MuddSummer亚型M蛋白可以包含如SEQ ID NO 1所示的氨基酸序列。在本申请中，所述溶瘤病毒的M蛋白可以包含如SEQ ID NO 3-5所示的氨基酸序列。
在本申请中，术语“G蛋白”通常指水VSV病毒的糖蛋白，也称包膜蛋白。术语“G蛋白”还包括其同源物、直系同源物、变体、功能活性片段等。在本申请中，野生型VSV病毒Indiana MuddSummer亚型G蛋白可以包含如SEQ ID NO 2所示的氨基酸序列。在本申请中，所述溶瘤病毒的糖蛋白G可以包含如SEQ ID NO 6-28所示的氨基酸序列。
在本申请中，术语“L蛋白”通常指VSV病毒RNA聚合酶蛋白。VSV病毒的L基因编码RNA poly E蛋白。术语“L蛋白”还包括其同源物，直系同源物、变体、功能活性片段等。
在本申请中，术语“N蛋白”通常指VSV病毒的核衣壳蛋白。术语“N蛋白”还包括其同源物，直系同源物、变体、功能活性片段等。
在本申请中，蛋白突变位点的表述通常由“氨基酸+氨基酸位数+(突变后的氨基酸)”来表述。在本申请中，所述突变可以包括但不限于氨基酸的增加、替换、缺失和/或删除。例如，术语“M51R”通常指第51位甲硫氨酸M突变为精氨酸R。
在本申请中，术语“氨基酸取代”通常是指用另一个氨基酸残基替代存在于亲本序列中的一个氨基酸残基。亲本序列中的氨基酸可以是例如经由化学肽合成或通过本领域已知的重组方法来取代。因此，“在xx位置处取代”通常是指用替代性氨基酸残基取代存在于位置xx处的氨基酸。在本申请中，所述氨基酸取代可以包括氨基酸突变。
在本申请中，术语“突变”通常是指改变野生型分子的核苷酸或氨基酸序列。氨基酸变化可以包括氨基酸的置换、删除、缺失、插入、添加、截短、或蛋白质的加工或切割。
在本申请中，术语“核酸分子”通常指任何长度的核苷酸。在本申请中，术语“核酸分子”可以编码所述溶瘤病毒包含的蛋白。在本申请中，所述核酸分子可以包含DNA和/或RNA。在某些情形下，所述RNA可以包含单链RNA(ssRNA)，也可以包含双链RNA(dsRNA)，单链RNA可以包含正义RNA，也可以包含反义RNA(anti-sense RNA),也可以包含双义RNA。
在本申请中，术语“表达载体”通常指一种核酸载体。在适当的条件下，其通常能够表达目标基因和/或目标蛋白。在本申请的某些实施方式中，所述表达载体包括用于表达病毒(例如，溶瘤病毒)的一种或多种组分的核酸分子。例如，所述表达载体可包括至少一个病毒基因组元件，并可以被包装入病毒或被包装为病毒粒子。
在本申请中，术语“病毒生产细胞”通常指可以或已经含有包括本申请所述的核酸分子或表达载体，或能够表达本申请所述溶瘤病毒的细胞、细胞系或细胞培养物。所述细胞可以包括单个宿主细胞的子代。所述细胞可以通过使用本申请所述的表达载体体外转染而得到。
在本申请中，术语“药物组合物”通常指以允许活性成分的生物学活性有效的形式存在的制剂，并且其不含有对所述制剂待施用的受试者有不可接受的毒性的另外成分。在某些实施方案中，这些制剂可以包含药物的活性组分以及药学上可接受的载剂。在某些实施方式中，所述药物产品包含肠胃外、经皮、腔内、动脉内、鞘内和/或鼻内施用或直接注射到组织中的药物产品。所述药物产品可以通过不同方式给药，例如静脉内、腹膜内、皮下、肌肉内、局部或真皮内施用。
在本申请中，术语“预防”通常是指通过预先采取某些措施而防止疾病或其一种或多种症状的产生和发作，复发，和/或扩散。在本申请中，术语“治疗”通常指消除或改善疾病，或与疾病相关的一种或多种症状。在某些实施方案中，治疗通常指向患有这种疾病的患者施用一种或多种药物使得疾病消除或缓解。在某些实施方案中，“治疗”可以是在特定疾病的症状发作后，在其他药物存在或不存在的情况下施用所述药物组合和/或药物产品。例如，使用本申请所述药物组合和/或药物产品防止肿瘤的产生，发展，复发和/或转移。
在本申请中，术语“肿瘤”通常是指任何新的病理性的组织增生。肿瘤可能是良性的，也可能是恶性的。在本申请中，所述肿瘤可以是实体瘤和/或血液瘤。用于研究时，可通过本领域技术人员熟知的方法从易于获得的资源中将这些组织分离出来。
发明详述
本申请提供了一种溶瘤病毒，所述溶瘤病毒是以野生型VSV病毒为基础，具体以VSV病毒印第安纳株，VSV病毒Indiana MuddSummer亚型株为基础，通过对其M蛋白和/或G蛋白的氨基酸序列上的位点进行突变获得的。其M蛋白的氨基酸序列如SEQ ID NO 1所示；其G蛋白的氨基酸序列如SEQ ID NO 2所示。在本申请中，所述M蛋白可以被改造，所述G蛋白也可以被改造。
本申请对所述VSV病毒进行了如下修饰，以获得溶瘤病毒：
所述溶瘤病毒包括M蛋白和G蛋白；所述M蛋白与SEQ ID NO 1所示的氨基酸序列相比，在第51位、第221位和第226位包含氨基酸取代；所述G蛋白与SEQ ID NO 2所示的氨基酸序列相比，包含至少一个氨基酸取代。
所述M蛋白的氨基酸取代包含在第51位的甲硫氨酸突变为精氨酸(M51R)；和/或，在第221位的缬氨酸突变为苯丙氨酸(V221F)；和/或，在第226位的丝氨酸突变为精氨酸(S226R)；例如，所述M蛋白包含如SEQ ID NO 5所示的氨基酸序列。在本申请中，所述M蛋白还可以包含其他位置的氨基酸取代。
在本申请中，所述G蛋白的氨基酸取代可以包含在第53位的缬氨酸突变为异亮氨酸(V53I)；和/或，在第141位的丙氨酸突变为缬氨酸(A141V)；和/或，在第172位的天冬氨酸突变为酪氨酸(D172Y)；和/或，在第217位的赖氨酸突变为谷氨酸(K217E)；和/或，在第232位的天冬氨酸突变为甘氨酸(D232G)；和/或，在第331位的缬氨酸突变为丙氨酸(V331A)；和/或，在第371位的缬氨酸突变为谷氨酸(V371E)；和/或，在第436位的甘氨酸突变为天冬氨酸(G436D)；和/或，在第438位的苏氨酸突变为丝氨酸(T438S)；和/或，在第453位的苯丙氨酸突变为亮氨酸(F453L)；和/或，在第471位的苏氨酸突变为异亮氨酸(T471I)；和/或，在第487位的酪氨酸突变为组氨酸(Y487H)；所述G蛋白可以包含如SEQ ID NO 17所示的氨基酸序列。
在本申请中，所述G蛋白可以包含第53位和第141位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位和第172位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位、第172位和第217位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位、第172位、第217位和第232位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位、第172位、第217位、第232位和第331位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位、第172位、第217位、第232位、第331位和第371位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位、第172位、第217位、第232位、第331位、第371位和第436位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位、第172位、第217位、第232位、第331位、第371位、第436位和第438位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位、第172位、第217位、第232位、第331位、第371位、第436位、第438位和第453位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位、第172位、第217位、第232位、第331位、第371位、第436位、第438位、第453位和第471位的氨基酸突变。
在本申请中，所述G蛋白可以包含第53位、第141位、第172位、第217位、第232位、第331位、第371位、第436位、第438位、第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第141位、第172位、第217位、第232位、第331位、第371位、第436位、第438位、第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第172位、第217位、第232位、第331位、第371位、第436位、第438位、第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第217位、第232位、第331位、第371位、第436位、第438位、第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第232位、第331位、第371位、第436位、第438位、第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第331位、第371位、第436位、第438位、第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第371位、第436位、第438位、第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第436位、第438位、第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第436位、第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第453位、第471位和第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第471位、第487位的氨基酸突变。
在本申请中，所述G蛋白可以包含第487位的氨基酸突变。
在本申请中，所述G蛋白还可以包含其他位置的氨基酸取代。
在某些实施方式中，所述G蛋白至少包含在保守区的一个或多个氨基酸取代。例如，所述保守区可以包含G蛋白的第437-461位氨基酸。在某些实施方式中，所述G蛋白至少包含在胞质域的截短区的一个或多个氨基酸取代。例如，所述胞质域的截短区可以包含G蛋白的第483-511位氨基酸。
在本申请中，所述G蛋白可以至少包含在第438、第453位、第471位和第487位的氨基酸取代。
所述溶瘤病毒还可以包含核酸分子和外源目的蛋白。该核酸分子可以包含编码所述具有氨基酸取代的M蛋白的核酸序列和编码所述具有氨基酸取代的G蛋白的核酸序列；该核酸分子还可以包含编码所述外源目的蛋白的核酸序列。进一步地，该核酸分子中所述编码外源目的蛋白的核酸序列位于所述编码所述具有氨基酸取代的M蛋白的核酸序列和所述编码所述具有氨基酸取代的G蛋白的核酸序列之间。再进一步地，该核酸分子编码所述溶瘤病毒的N蛋白(核蛋白N)、L蛋白(大聚合酶蛋白L)和P蛋白(磷酸蛋白P)。
在本申请中，可以通过病毒包装过程和病毒拯救过程获得本申请所述的溶瘤病毒。具体过程可以包括用表达T7 RNA聚合酶的痘病毒vTF7-3感染接种BSR-T7细胞，使用分别克隆VSV N、VSV P、VSV L基因的表达质粒和骨架质粒进行lipofectamine转染，获得目的溶瘤病毒。
本申请还提供了一种溶瘤病毒表达载体、一种病毒生产细胞以及一种药物组合物。
所述溶瘤病毒表达载体可以包含编码所述溶瘤病毒的M蛋白和G蛋白的核酸序列；所述溶瘤病毒表达载体还可以包含编码所述溶瘤病毒的N蛋白、P蛋白和L蛋白的核酸序列。
所述病毒生产细胞能够产生上述的溶瘤病毒；所述病毒生产细胞可以包含BSR-T7细胞。
所述药物组合物包括上述的溶瘤病毒，以及任选地药学上可接受的载剂。
在某些实施方案中，所述药物组合物可以包括一种或多种(药学上有效的)佐剂、稳定剂、赋形剂、稀释剂、增溶剂、表面活性剂、乳化剂和/或防腐剂的合适的制剂。药物组合物的可接受成分在所用剂量和浓度下优选对接受者无毒。本申请的药物组合物包括但不限于液体、冷冻和冻干组合物。
在某些实施方案中，所述药学上可接受的载剂可以包括与药物给药相容的任何和所有的溶剂、分散介质、包衣、等渗剂和吸收延迟剂，通常安全、无毒。
在某些实施方案中，所述药物组合物可以包含肠胃外、经皮、腔内、动脉内、鞘内和/或鼻内施用或直接注射到组织中。例如，所述药物组合物可以通过输注或注射施用于患者或者受试者。在某些实施方案中，所述药物组合物的施用可以通过不同的方式进行，例如静脉内、腹膜内、皮下、肌肉内、局部或真皮内施用。在某些实施方案中，所述药物组合物可以不间断施用。所述不间断(或连续)施用可以通过患者佩戴的小泵系统来实现，以测量流入患者体内的治疗剂，如WO2015/036583所述。
此外，本申请还提供了上述溶瘤病毒的制备方法，该制备方法可以包括溶瘤病毒表达载体、病毒生产细胞和/或药物组合物的制备方法。任何适于生产溶瘤病毒的方法都可用于产生本申请的溶瘤病毒。例如，可以向细胞中加入表达T7 RNA聚合酶的痘病毒进行转染，加入表达所述溶瘤病毒N蛋白、L蛋白和P蛋白的质粒以及骨架质粒进行转染，通过病毒拯救过程获得本申请的溶瘤病毒。
本申请还提供了上述溶瘤病毒、溶瘤病毒表达载体、病毒生产细胞和/或药物组合物在制备用于预防和/或治疗疾病和/或病症的药物中的用途。
本申请提供的溶瘤病毒均对LLC细胞、MC38细胞以及Hela细胞具有较好的侵染能力，尤其是编号为JBS104-JBS126的溶瘤病毒，在编号为JBS103的溶瘤病毒的基础上，对溶瘤病毒的G蛋白上的氨基酸进行定点突变，从而进一步提高了溶瘤病毒对LLC细胞、MC38细胞以及Hela细胞的侵染能力。同时，上述制备的溶瘤病毒对正常细胞MEF细胞的侵染能力均较差，说明本申请制备的溶瘤病毒能够较好的用于肿瘤、癌症等细胞的侵染，同时不会损伤正常细胞，具有广泛的应用前景。
本申请提供的溶瘤病毒均对LLC细胞、MC38细胞以及4T1细胞具有较好的体外杀伤能力，尤其是编号为JBS104-JBS126的溶瘤病毒，在编号为JBS103的溶瘤病毒的基础上，对溶瘤病毒的G蛋白上的氨基酸进行定点突变，从而进一步提高了溶瘤病毒对LLC细胞、MC38细胞以及4T1细胞的体外杀伤能力。同时，上述制备的溶瘤病毒对MEF细胞几乎无影响，说明本申请制备的溶瘤病毒能够较好的用于肿瘤、癌症等非正常细胞的损伤和杀死，同时不会损伤正常细胞。
编号为JBS101和JBS102的溶瘤病毒在LLC细胞、MC38细胞以及Hela细胞内均不容易清除。相对而言，编号为JBS103的溶瘤病毒在LLC细胞、MC38细胞以及Hela细胞内更容易清除。而本申请编号为JBS104-JBS126的溶瘤病毒，在编号为JBS103的溶瘤病毒的基础上，进一步对溶瘤病毒的G蛋白上的氨基酸进行定点突变，从而使得溶瘤病毒在LLC细胞、MC38细胞以及Hela细胞内更加不容易清除，进一步保证溶瘤病毒能够在LLC细胞、MC38细胞以及Hela细胞内更好的发挥侵染和杀伤能力；同时，本申请提供的溶瘤病毒在MEF细胞内更加容易清除，进一步保证了MEF细胞的安全性，从而提高了溶瘤病毒的安全性。
不欲被任何理论所限，下文中的实施例仅仅是为了阐释本申请发明的各个技术方案，而不用于限制本申请发明的范围。
制备例
制备例1-3
制备例1-3分别提供了一种溶瘤病毒，其不同之处主要在于野生型溶瘤病毒M蛋白上氨基酸定点突变的位点。各制备例对应的溶瘤病毒的构建方法具体如下：
(1)构建质粒
以pRV-core质粒(BioVector NTCC质粒载体菌种细胞基因保藏中心)为模板，利用PCR技术引入如表1所示的突变位点。将pRV-core质粒分别与载有各突变位点的引物进行PCR；然后将PCR产物进行1％琼脂糖凝胶电泳；再利用凝胶回收试剂盒进行割胶回收，从而获得具有M蛋白不同突变位点的质粒，获得构建好的质粒pRV-core Mut。
表1制备例1-3中溶瘤病毒M蛋白的突变情况表
(2)病毒拯救
利用磷酸钙转染试剂盒(Thermo Fisher Scientific)，通过细胞转染技术，将构建好的质粒pRV-core Mut转染至BSR-T7细胞(购自ATCC，美国菌种保藏中心，又称美国模式菌种收集中心)。
按照pRV-core Mut、pP、pN、pL的质量比例为10:5:4:1，将四种质粒混合，质粒总量为5μg；使用200μl opti-MEM培养基(Thermo Fisher Scientific)稀释质粒，并加入7.5μl转染试剂Plus Reagent(Life Technologies)，获得转染质粒预混液；其中，pP(载有棒状病毒磷酸蛋白基因的质粒)、pN(载有棒状病毒核蛋白基因的质粒)、pL(载有棒状病毒聚合酶蛋白基因的质粒)；pN、pP、pL三种质粒对应的母体载体均为pCAGGS(购自ATCC)；
利用200μl opti-MEM培养基稀释Lipofectamine LTX 10μl(Thermo FisherScientific)，获得LTX混合液；
按照Lipofectamine LTX使用说明书所述的方法进行质粒转染，6h后利用PBS清洗BSR-T7细胞两次，进一步接种于10％胎牛血清的DMEM培养基(Thermo Fisher Scientific)中培养3天；
将培养BSR-T7细胞获得的的细胞上清转移到Vero细胞(Thermo FisherScientific)中，并将Vero细胞置于37℃的环境条件下培养3天，荧光显微镜观察细胞中绿色荧光确定病毒拯救的情况；进一步将拯救的突变棒状病毒库，通过Vero细胞传代，在建立的噬菌斑筛选系统中挑取单克隆病毒株。
(3)M蛋白基因测序。利用Trizol试剂盒抽提病毒基因组RNA，用随机引物进行逆转录反应，用针对M蛋白基因序列设计的引物对逆转录出来的cDNA进行PCR；
引物序列为：
5'-AAAAAAGTAACAGATATCAC-3'；
5'-ACATTTTTCCAGTTTCCTTTTTGG-3'。
产物经1％琼脂糖凝胶电泳后回收，并送往测序公司进行测序。测序结果如表1所示。
制备例4-26
制备例4-26分别提供了一种溶瘤病毒，其不同之处主要在于野生型溶瘤病毒M蛋白和G蛋白上氨基酸定点突变的位点，其中M蛋白的突变位点与制备例3中相应的突变位点相同。各制备例对应的溶瘤病毒的构建方法同制备例1-3中所述的构建方法，不同之处在于：
步骤(1)的构建质粒中利用PCR技术引入如表2所示的突变位点；
步骤(3)为G蛋白基因测序。利用Trizol试剂盒抽提病毒基因组RNA，用随机引物进行逆转录反应，用针对G蛋白基因序列设计的引物对逆转录出来的cDNA进行PCR；
引物序列为：
5’-CCATGGCCTGTTGCTCCACCAGCTT-3’；
5’-AAGCTTTCAGCAGTGGCTCACAGCAG-3’。
产物经1％琼脂糖凝胶电泳后回收，并送往测序公司进行测序。测序结果如表2所示。
表2制备例4-26中溶瘤病毒G蛋白的突变情况表
制备例27
本制备例提供了利用上述实施例1-26制备的溶瘤病毒的包装过程，具体包括以下步骤：
1)利用表达T7 RNA聚合酶的痘病毒vTF7-3(BioVector NTCC质粒载体菌种细胞基因保藏中心)感染接种BSR-T7细胞(购自ATCC)
具体步骤如下：将BSR-T7细胞铺在6孔板上，控制每孔细胞量达到3×105个；铺板14-16h后加入表达T7 RNA聚合酶的痘病毒vTF7-3，进行痘病毒vTF7-3对BSR-T7细胞的感染；感染6h后，使用DPBS缓冲液(Thermo Fisher Scientific)漂洗一次BSR-T7细胞，进行转染。
2)转染过程
具体包括以下步骤：按照pRV-core Mut、pP、pN、pL的质量比例为10:5:4:1，将四种质粒混合，质粒总量为5μg；使用200μl opti-MEM培养基(Thermo Fisher Scientific)稀释质粒，并加入7.5μl转染试剂Plus Reagent(Life Technologies)，获得转染质粒预混液；其中，pP(载有棒状病毒磷酸蛋白基因的质粒)、pN(载有棒状病毒核蛋白基因的质粒)、pL(载有棒状病毒聚合酶蛋白基因的质粒)；pN、pP、pL三种质粒对应的母体载体均为pCAGGS(购自ATCC)；
利用200μl opti-MEM培养基稀释Lipofectamine LTX 10μl(Thermo FisherScientific)，获得LTX混合液；
将LTX混合液200μl与转染质粒预混液200μl混合，室温孵育15min，获得LTX-DNA混合液；
将步骤1)的6孔板中的DPBS缓冲液换成Opti-MEM培养基，将LTX-DNA混合液逐滴加入培养BSR-T7细胞的6孔板中，轻轻摇动6孔板，使LTX-DNA混合液均匀分布在6孔板内；转染6-8h后，吸去转染试剂，加入3ml新鲜的完全培养基(Thermo Fisher Scientific)；72h后收获BSR-T7细胞的细胞上清，使用0.22μm滤器进行过滤，获得制备例1-26各自对应的溶瘤病毒滴。
实施例
实施例1
本实施例分别利用制备例1-26制得的溶瘤病毒以及野生型溶瘤病毒对不同细胞的侵染能力进行检测。检测方法为TCID50检测法，即在不同细胞的培养液中，分别加入制备例1-26以及野生型溶瘤病毒各200pfu，检测各溶瘤病毒产生的半数组织培养感染剂量(Tissue culture infective dose，TCID50)。所检测的细胞包括：LLC细胞(鼠源非小细胞肺癌细胞)、MC38细胞(小鼠结肠癌细胞)、Hela细胞(人宫颈癌细胞)、MEF细胞(人成纤维细胞)。具体检测方法为：
(1)分别在6孔培养板中加入Vero(LLC/MC38/Hela/MEF)细胞悬液3mL，使细胞量达到4×105个/孔，每种Vero细胞共6个孔，MEF细胞设置2个孔，作为对照；将6孔培养板置于37℃、5％CO2的环境条件下培养16h；
(2)向6孔培养板的每个孔中加入制备例制得的溶瘤病毒200pfu，在24h收获MEF细胞以及每种Vero细胞的上清100μl；将收获的分别加入96孔培养板的孔中，使各种细胞的细胞量达到1×104个/ml，将96孔培养板置于37℃、5％CO2的环境条件下培养16h；
(3)在1.5ml EP管中将步骤(2)收获的上清作连续10倍的稀释，从10-1～10-11，共11个滴度；将稀释好的上清接种到96孔培养板中，每一稀释度接种一列共8孔，每孔接种100μl；
(4)48h后观察每孔细胞的荧光情况，有荧光则记为此孔被感染；按Karber法计算TCID50。
检测结果如图1-图4所示，其中，横坐标0表示野生型溶瘤病毒；横坐标1-26分别表示制备例1-26制备的溶瘤病毒；纵坐标Log 10TCID50表示利用Karber法计算得出的TCID50值，Log 10TCID50的值越大，表示溶瘤病毒对该细胞的侵染能力越好；Log 10TCID50的值越小，表示溶瘤病毒对该细胞的侵染能力越差。
图1为本申请制备的溶瘤病毒以及野生型溶瘤病毒对LLC细胞的侵染能力的检测结果。
图2为本申请制备的溶瘤病毒以及野生型溶瘤病毒对MC38细胞的侵染能力的检测结果。
图3为本申请制备的溶瘤病毒以及野生型溶瘤病毒对Hela细胞的侵染能力的检测结果。
图4为本申请制备的溶瘤病毒以及野生型溶瘤病毒对MEF细胞的侵染能力的检测结果。
通过上述附图可知，本申请制备例1-26制备的溶瘤病毒均对LLC细胞、MC38细胞以及Hela细胞具有较好的侵染能力，尤其是制备例4-26，在制备例3的基础上进一步对溶瘤病毒的G蛋白上的氨基酸进行定点突变，从而进一步提高了溶瘤病毒对LLC细胞、MC38细胞以及Hela细胞的侵染能力。同时，上述制备的溶瘤病毒均对MEF细胞的侵染能力较差，说明本申请制备的溶瘤病毒能够较好的用于肿瘤、癌症等细胞的侵染，同时不会损伤正常细胞，具有广泛的应用前景。
实施例2
本实施例分别利用制备例1-26制得的溶瘤病毒以及野生型溶瘤病毒对不同细胞进行体外杀伤试验检测。检测方法为MTT检测法，即在不同细胞的培养液中，分别加入制备例1-26以及野生型溶瘤病毒各200pfu，24h后进行利用MTT检测法检测细胞活性。所检测的细胞包括：LLC细胞、MC38细胞、MEF细胞、4T1细胞(小鼠乳腺细胞)。具体检测方法为：
(1)分别在96孔培养板中加入Vero(LLC/MC38/4T1/MEF)细胞悬液100μl，使细胞量达到1×104个/孔，将96孔培养板置于37℃、5％CO2的环境条件下培养16h；
(2)将制备例制得的溶瘤病毒稀释到MOI(感染复数)分别为0.001、0.01、0.1、1.0，将每一稀释梯度的溶瘤病毒分别接种到步骤(1)的96孔培养板上，每一稀释梯度接种4个孔，每孔100μl，将96孔培养板置于37℃、5％CO2的环境条件下培养40h；
(3)将步骤(2)的96孔培养板中的细胞上清取出，并向96孔培养板中加入新鲜培养基和MTT溶液，加入量为20μL/孔，将96孔培养板置于37℃、5％CO2的环境条件下培养4h；
(4)将96孔培养板于室温下离心5min，设置转速为2500rpm/min，用1mL一次性无菌注射器轻轻吸掉上清；然后向96孔培养板的每个孔中加入DMSO，加入量为100ul/孔，37℃的环境条件下放置10min。使用多功能酶标仪，震荡2min，在570nm或490nm波长下测定96孔培养板上各孔的OD值。
检测结果如图5-图8所示，其中，横坐标0表示野生型溶瘤病毒；横坐标1-26分别表示制备例1-26制备的溶瘤病毒；纵坐标OD570表示细胞的OD值，OD570的值越大，表示溶瘤病毒对该细胞的杀伤能力越差；OD570的值越小，表示溶瘤病毒对该细胞的杀伤能力越好。
图5为本申请制备的溶瘤病毒以及野生型溶瘤病毒对LLC细胞的体外杀伤能力的检测结果。
图6为本申请制备的溶瘤病毒以及野生型溶瘤病毒对MC38细胞的体外杀伤能力的检测结果。
图7为本申请制备的溶瘤病毒以及野生型溶瘤病毒对4T1细胞的体外杀伤能力的检测结果。
图8为本申请制备的溶瘤病毒以及野生型溶瘤病毒对MEF细胞的体外杀伤能力的检测结果。
通过上述附图可知，本申请制备例1-26制备的溶瘤病毒均对LLC细胞、MC38细胞以及4T1细胞具有较好的体外杀伤能力，尤其是制备例4-26，在制备例3的基础上进一步对溶瘤病毒的G蛋白上的氨基酸进行定点突变，从而进一步提高了溶瘤病毒对LLC细胞、MC38细胞以及4T1细胞的体外杀伤能力。同时，上述制备的溶瘤病毒对MEF细胞几乎无影响，说明本申请制备的溶瘤病毒能够较好的用于肿瘤、癌症等非正常细胞的损伤和杀死，同时不会损伤正常细胞。
虽然野生型溶瘤病毒对LLC细胞、MC38细胞以及4T1细胞已具有较好的体外杀伤能力，但其在损伤和杀死上述细胞的同时，还会很大程度地损伤和杀死MEF细胞，限制了野生型溶瘤病毒的临床应用。因此，本申请对野生型溶瘤病毒的改造，在保证溶瘤病毒对正常细胞的安全性的同时，还保证了溶瘤病毒对肿瘤、癌症细胞的杀伤能力，具有广泛的临床应用前景。
实施例3
本实施例对制备例1-26制得的溶瘤病毒以及野生型溶瘤病毒在不同细胞内的清除难易度进行检测。检测指标为基因IFN-β的表达情况。基因IFN-β是细胞产生的具有广泛的抗病毒、抗肿瘤和免疫调节作用的可溶性糖蛋白的基因，通过基因IFN-β的表达情况可判断细胞对溶瘤病毒的清除能力：当基因IFN-β的表达较高时，表明溶瘤病毒在细胞内容易清除；当基因IFN-β的表达较低时，表示溶瘤病毒在细胞内不容易清除。所检测的细胞包括：LLC细胞、MC38细胞、Hela细胞、MEF细胞。具体检测方法为：
(1)分别在96孔培养板中加入Vero(LLC/MC38/Hela/MEF)细胞以及MEF细胞悬液100μl，使细胞量达到1×104个/孔，将96孔培养板置于37℃、5％CO2的环境条件下培养16h；
(2)将制备例制得的溶瘤病毒稀释到MOI(感染复数)分别为0.001、0.01、0.1、1.0，将每一稀释梯度的溶瘤病毒分别接种到步骤(1)的96孔培养板上，每一稀释梯度接种4个孔，每孔100μl，将96孔培养板置于37℃、5％CO2的环境条件下培养40h；
(3)将步骤(2)培养获得的各组细胞破碎，并利用TRIzol(Invitrogen)从各细胞中提取总RNA，利用PrimeScript RT Reagent Kit with DNA Eraser(Takara)反转录试剂盒逆转录成cDNA，并用LightCycler 480SYBR Green I Master(Roche)染料进行染色，在LightCycler 480定量PCR仪上检测各个基因的Ct值。用ΔΔCt法计算目的基因IFN-β的相对表达量。
检测结果如图9-图12所示，其中，横坐标0表示野生型溶瘤病毒；横坐标1-26分别表示制备例1-26制备的溶瘤病毒；纵坐标IFN-β水平表示IFN-β基因的表达情况，IFN-β水平的值越大，表示溶瘤病毒在该细胞内越容易清除；IFN-β水平的值越小，表示溶瘤病毒在该细胞内越不容易清除。
图9为本申请制备的溶瘤病毒以及野生型溶瘤病毒在LLC细胞内的清除难易度的检测结果。
图10为本申请制备的溶瘤病毒以及野生型溶瘤病毒在MC38细胞的清除难易度的检测结果。
图11为本申请制备的溶瘤病毒以及野生型溶瘤病毒在Hela细胞的清除难易度的检测结果。
图12为本申请制备的溶瘤病毒以及野生型溶瘤病毒在MEF细胞的清除难易度的检测结果。
通过上述附图可知，本申请制备例1-2在LLC细胞、MC38细胞以及Hela细胞内均不容易清除。相对制备例1和制备例2而言，制备例3在LLC细胞、MC38细胞以及Hela细胞内更容易清除。本申请制备例4-26，在制备例3的基础上进一步对溶瘤病毒的G蛋白上的氨基酸进行定点突变，从而使得溶瘤病毒在LLC细胞、MC38细胞以及Hela细胞内更加不容易清除的程度，进一步保证溶瘤病毒能够在LLC细胞、MC38细胞以及Hela细胞内更好的发挥侵染和杀伤的能力；同时，使得溶瘤病毒在MEF细胞内更加容易清除，进一步保证了MEF细胞的安全性，从而提高了溶瘤病毒的安全性。
前述详细说明是以解释和举例的方式提供的，并非要限制所附权利要求的范围。目前本申请所列举的实施方式的多种变化对本领域普通技术人员来说是显而易见的，且保留在所附的权利要求和其等同方式的范围内。
序列表
<110> 上海荣瑞医药科技有限公司
<120> 一种溶瘤病毒及其用途
<160> 28
<170> SIPOSequenceListing 1.0
<210> 1
<211> 229
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Met Ser Ser Leu Leu Leu Ile Leu Gly Leu Leu Gly Leu Gly Leu Leu
1 5 10 15
Ser Leu Leu Leu Gly Ile Ala Pro Pro Pro Thr Gly Gly Ala Thr Ala
20 25 30
Met Gly Thr Ala Pro Ser Ala Pro Ile Ala Leu Ser Thr Pro Gly Val
35 40 45
Ala Gly Met Ala Thr His Ala Pro Ala Gly Leu Ala Thr Gly Leu Pro
50 55 60
Pro Pro Thr Val Leu Met Thr Val Ala Ser Ala Ala Pro Pro Ala Thr
65 70 75 80
Thr Ser Ala Val Ala Ala Ala Val Ser His Thr Ala His Met Thr Ile
85 90 95
Gly Met Ala Gly Leu Ala Pro Pro Thr Leu Ile Leu Ala Pro Leu Gly
100 105 110
Ser Ser Ala Leu Leu Ala Thr Pro Ala Val Leu Ala Ala Gly Gly Gly
115 120 125
Pro Gly Thr His Ala His Cys Gly Gly Ala Ala Thr Leu Pro His Ala
130 135 140
Met Gly Leu Thr Pro Pro Met Leu Ala Val Pro Gly His Pro Ala Ala
145 150 155 160
Pro Pro Ala Ile Gly Leu Thr Leu Gly Thr Ile Gly Leu Thr Met Thr
165 170 175
Ile Thr Ala Ala Gly Ser Leu Gly Ala Ala Pro Met Ile Thr Ala His
180 185 190
Pro Ala Ser Ser Leu Pro Ser Ala Pro Ala Gly Leu Ala Leu Met Pro
195 200 205
Gly Leu Ile Val Gly Leu Leu Ala Ser Gly Ala Thr Val Leu Ala Ser
210 215 220
Val Ser His Pro Leu
225
<210> 2
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 2
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 3
<211> 229
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Met Ser Ser Leu Leu Leu Ile Leu Gly Leu Leu Gly Leu Gly Leu Leu
1 5 10 15
Ser Leu Leu Leu Gly Ile Ala Pro Pro Pro Thr Gly Gly Ala Thr Ser
20 25 30
Met Gly Thr Ala Pro Ser Ala Pro Ile Ala Leu Ser Thr Pro Gly Val
35 40 45
Ala Gly Ala Ala Thr Thr Ala Pro Ala Gly Leu Ala Thr Gly Leu Pro
50 55 60
Pro Pro Thr Val Leu Met Thr Val Ala Ser Ala Ala Pro Pro Ala Thr
65 70 75 80
Thr Ser Ala Val Ala Ala Ala Val Ser His Thr Ala His Met Thr Ile
85 90 95
Gly Met Ala Gly Leu Ala Pro Pro Thr Leu Ile Leu Ala Pro Leu Gly
100 105 110
Ser Ser Ala Leu Leu Ala Thr Pro Ala Val Leu Ala Ala Gly Gly Gly
115 120 125
Pro Gly Thr His Ala His Cys Gly Gly Ala Ala Thr Leu Pro His Ala
130 135 140
Met Gly Leu Thr Pro Pro Met Leu Ala Val Pro Gly His Pro Ala Ala
145 150 155 160
Pro Pro Ala Ile Gly Leu Thr Leu Gly Thr Ile Gly Leu Thr Met Thr
165 170 175
Ile Thr Ala Ala Gly Ser Leu Gly Ala Ala Pro Met Ile Thr Ala His
180 185 190
Pro Ala Ser Ser Leu Pro Ser Ala Pro Ala Gly Leu Ala Leu Met Pro
195 200 205
Gly Leu Ile Val Gly Leu Leu Ala Ser Gly Ala Thr Val Leu Ala Ser
210 215 220
Ile Ser His Pro Leu
225
<210> 4
<211> 229
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 4
Met Ser Ser Leu Leu Leu Ile Leu Gly Leu Leu Gly Leu Gly Leu Leu
1 5 10 15
Ser Leu Leu Leu Gly Ile Ala Pro Pro Pro Thr Gly Gly Ala Thr Ser
20 25 30
Met Gly Thr Ala Pro Ser Ala Pro Ile Ala Leu Ser Thr Pro Gly Val
35 40 45
Ala Gly Ala Ala Thr Thr Ala Pro Ala Gly Leu Ala Thr Gly Leu Pro
50 55 60
Pro Pro Thr Val Leu Met Thr Val Ala Ser Ala Ala Pro Pro Ala Thr
65 70 75 80
Thr Ser Ala Val Ala Ala Ala Val Ser His Thr Ala His Met Thr Ile
85 90 95
Gly Met Ala Gly Leu Ala Pro Pro Thr Leu Ile Leu Ala Pro Leu Gly
100 105 110
Ser Ser Ala Leu Leu Ala Thr Pro Ala Val Leu Ala Ala Gly Gly Gly
115 120 125
Pro Gly Thr His Ala His Cys Gly Gly Ala Ala Thr Leu Pro His Ala
130 135 140
Met Gly Leu Thr Pro Pro Met Leu Ala Val Pro Gly His Pro Ala Ala
145 150 155 160
Pro Pro Ala Ile Gly Leu Thr Leu Gly Thr Ile Gly Leu Thr Met Thr
165 170 175
Ile Thr Ala Ala Gly Ser Leu Gly Ala Ala Pro Met Ile Thr Ala His
180 185 190
Pro Ala Ser Ser Leu Pro Ser Ala Pro Ala Gly Leu Ala Leu Met Pro
195 200 205
Gly Leu Ile Val Gly Leu Leu Ala Ser Gly Ala Thr Pro Leu Ala Ser
210 215 220
Ile Ser His Pro Leu
225
<210> 5
<211> 229
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 5
Met Ser Ser Leu Leu Leu Ile Leu Gly Leu Leu Gly Leu Gly Leu Leu
1 5 10 15
Ser Leu Leu Leu Gly Ile Ala Pro Pro Pro Thr Gly Gly Ala Thr Ser
20 25 30
Met Gly Thr Ala Pro Ser Ala Pro Ile Ala Leu Ser Thr Pro Gly Val
35 40 45
Ala Gly Ala Ala Thr Thr Ala Pro Ala Gly Leu Ala Thr Gly Leu Pro
50 55 60
Pro Pro Thr Val Leu Met Thr Val Ala Ser Ala Ala Pro Pro Ala Thr
65 70 75 80
Thr Ser Ala Val Ala Ala Ala Val Ser His Thr Ala His Met Thr Ile
85 90 95
Gly Met Ala Gly Leu Ala Pro Pro Thr Leu Ile Leu Ala Pro Leu Gly
100 105 110
Ser Ser Ala Leu Leu Ala Thr Pro Ala Val Leu Ala Ala Gly Gly Gly
115 120 125
Pro Gly Thr His Ala His Cys Gly Gly Ala Ala Thr Leu Pro His Ala
130 135 140
Met Gly Leu Thr Pro Pro Met Leu Ala Val Pro Gly His Pro Ala Ala
145 150 155 160
Pro Pro Ala Ile Gly Leu Thr Leu Gly Thr Ile Gly Leu Thr Met Thr
165 170 175
Ile Thr Ala Ala Gly Ser Leu Gly Ala Ala Pro Met Ile Thr Ala His
180 185 190
Pro Ala Ser Ser Leu Pro Ser Ala Pro Ala Gly Leu Ala Leu Met Pro
195 200 205
Gly Leu Ile Val Gly Leu Leu Ala Ser Gly Ala Thr Pro Leu Ala Ser
210 215 220
Ile Ala His Pro Leu
225
<210> 6
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 7
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 7
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 8
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 8
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 9
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 9
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 10
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 10
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 11
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 11
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 12
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 13
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 14
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 15
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 16
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile Thr Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 17
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Ile Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 18
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Val Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 19
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Thr Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 20
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Gly Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 21
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 21
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Gly Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 22
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Ala Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 23
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 23
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Gly Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 24
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Ala Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 25
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 25
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Ser Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 26
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Leu Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 27
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Ile Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
<210> 28
<211> 511
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Met Leu Cys Leu Leu Thr Leu Ala Pro Leu Pro Ile Gly Val Ala Cys
1 5 10 15
Leu Pro Thr Ile Val Pro Pro His Ala Gly Leu Gly Ala Thr Leu Ala
20 25 30
Val Pro Ser Ala Thr His Thr Cys Pro Ser Ser Ser Ala Leu Ala Thr
35 40 45
His Ala Ala Leu Val Gly Thr Ala Leu Gly Val Leu Met Pro Leu Ser
50 55 60
His Leu Ala Ile Gly Ala Ala Gly Thr Met Cys His Ala Ser Leu Thr
65 70 75 80
Val Thr Thr Cys Ala Pro Ala Thr Thr Gly Pro Leu Thr Ile Thr His
85 90 95
Ser Ile Ala Ser Pro Thr Pro Ser Val Gly Gly Cys Leu Gly Ser Ile
100 105 110
Gly Gly Thr Leu Gly Gly Thr Thr Leu Ala Pro Gly Pro Pro Pro Gly
115 120 125
Ser Cys Gly Thr Ala Thr Val Thr Ala Ala Gly Ala Ala Ile Val Gly
130 135 140
Val Thr Pro His His Val Leu Val Ala Gly Thr Thr Gly Gly Thr Val
145 150 155 160
Ala Ser Gly Pro Ile Ala Gly Leu Cys Ser Ala Ala Ile Cys Pro Thr
165 170 175
Val His Ala Ser Thr Thr Thr His Ser Ala Thr Leu Val Leu Gly Leu
180 185 190
Cys Ala Ser Ala Leu Ile Ser Met Ala Ile Thr Pro Pro Ser Gly Ala
195 200 205
Gly Gly Leu Ser Ser Leu Gly Leu Leu Gly Thr Gly Pro Ala Ser Ala
210 215 220
Thr Pro Ala Thr Gly Thr Gly Ala Leu Ala Cys Leu Met Gly Thr Cys
225 230 235 240
Leu His Thr Gly Val Ala Leu Pro Ser Gly Val Thr Pro Gly Met Ala
245 250 255
Ala Leu Ala Leu Pro Ala Ala Ala Ala Pro Pro Gly Cys Pro Gly Gly
260 265 270
Ser Ser Ile Ser Ala Pro Ser Gly Thr Ser Val Ala Val Ser Leu Ile
275 280 285
Gly Ala Val Gly Ala Ile Leu Ala Thr Ser Leu Cys Gly Gly Thr Thr
290 295 300
Ser Leu Ile Ala Ala Gly Leu Pro Ile Ser Pro Val Ala Leu Ser Thr
305 310 315 320
Leu Ala Pro Leu Ala Pro Gly Thr Gly Pro Val Pro Thr Ile Ile Ala
325 330 335
Gly Thr Leu Leu Thr Pro Gly Thr Ala Thr Ile Ala Val Ala Ile Ala
340 345 350
Ala Pro Ile Leu Ser Ala Met Val Gly Met Ile Ser Gly Thr Thr Thr
355 360 365
Gly Ala Val Leu Thr Ala Ala Thr Ala Pro Thr Gly Ala Val Gly Ile
370 375 380
Gly Pro Ala Gly Val Leu Ala Thr Ser Ser Gly Thr Leu Pro Pro Leu
385 390 395 400
Thr Met Ile Gly His Gly Met Leu Ala Ser Ala Leu His Leu Ser Ser
405 410 415
Leu Ala Gly Val Pro Gly His Pro His Ile Gly Ala Ala Ala Ser Gly
420 425 430
Leu Pro Ala Gly Gly Thr Leu Pro Pro Gly Ala Thr Gly Leu Ser Leu
435 440 445
Ala Pro Ile Gly Pro Val Gly Gly Thr Pro Ser Ser Thr Leu Ser Ser
450 455 460
Ile Ala Ser Pro Pro Pro Thr Ile Gly Leu Ile Ile Gly Leu Pro Leu
465 470 475 480
Val Leu Ala Val Gly Ile His Leu Cys Ile Leu Leu Leu His Thr Leu
485 490 495
Leu Ala Gly Ile Thr Thr Ala Ile Gly Met Ala Ala Leu Gly Leu
500 505 510
Claims (26)
1.溶瘤病毒，其特征在于，所述溶瘤病毒包括M蛋白和G蛋白；所述M蛋白与SEQ ID NO 1所示的氨基酸序列相比，包含在第51位、第221位和第226位的氨基酸取代；所述G蛋白与SEQID NO 2所示的氨基酸序列相比，包含至少一个氨基酸取代。
2.根据权利要求1所述的溶瘤病毒，其特征在于：所述M蛋白的氨基酸取代包含在第51位的甲硫氨酸突变为精氨酸（M51R）；和/或，在第221位的缬氨酸突变为苯丙氨酸（V221F）；和/或，在第226位的丝氨酸突变为精氨酸（S226R）。
3.根据权利要求1-2中任一项所述的溶瘤病毒，其特征在于：所述M蛋白包含如SEQ IDNO 5所示的氨基酸序列。
4.根据权利要求1-3中任一项所述的溶瘤病毒，其特征在于：所述G蛋白与SEQ ID NO 2所示的氨基酸序列相比，包含以下一个或多个位点的氨基酸取代：第438位、第453位、第471位和第487位。
5.根据权利要求1-3中任一项所述的溶瘤病毒，其特征在于：所述G蛋白与SEQ ID NO 2所示的氨基酸序列相比，包含以下一个或多个位点的氨基酸取代：第53位、第141位、第172位、第217位、第232位、第331位、第371位、第436位、第438位、第453位、第471位和第487位。
6.根据权利要求1-5中任一项所述的溶瘤病毒，其特征在于：所述G蛋白的氨基酸取代包含在第53位的缬氨酸突变为异亮氨酸（V53I）；和/或，在第141位的丙氨酸突变为缬氨酸（A141V）；和/或，在第172位的天冬氨酸突变为酪氨酸（D172Y）；和/或，在第217位的赖氨酸突变为谷氨酸（K217E）；和/或，在第232位的天冬氨酸突变为甘氨酸（D232G）；和/或，在第331位的缬氨酸突变为丙氨酸（V331A）；和/或，在第371位的缬氨酸突变为谷氨酸（V371E）；和/或，在第436位的甘氨酸突变为天冬氨酸（G436D）；和/或，在第438位的苏氨酸突变为丝氨酸（T438S）；和/或，在第453位的苯丙氨酸突变为亮氨酸（F453L）；和/或，在第471位的苏氨酸突变为异亮氨酸（T471I）；和/或，在第487位的酪氨酸突变为组氨酸（Y487H）。
7.根据权利要求1-6中任一项所述的溶瘤病毒，其特征在于：所述G蛋白的氨基酸取代选自下组中的任一项：
1）所述G蛋白的氨基酸取代包含V53I；
2）所述G蛋白的氨基酸取代包含V53I和A141V；
3）所述G蛋白的氨基酸取代包含V53I、A141V和D172Y；
4）所述G蛋白的氨基酸取代包含V53I、A141V、D172Y和K217E；
5）所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E和D232G；
6）所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G和V331A；
7）所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A和V371E；
8）所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E和G436D；
9）所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D和T438S；
10）所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S和F453L；
11）所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L和T471I；
12）所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H；
13）所述G蛋白的氨基酸取代包含A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H；
14）所述G蛋白的氨基酸取代包含D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H；
15）所述G蛋白的氨基酸取代包含K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H；
16）所述G蛋白的氨基酸取代包含D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H；
17）所述G蛋白的氨基酸取代包含V331A、V371E、G436D、T438S、F453L、T471I和Y487H；
18）所述G蛋白的氨基酸取代包含V371E、G436D、T438S、F453L、T471I和Y487H；
19）所述G蛋白的氨基酸取代包含G436D、T438S、F453L、T471I和Y487H；
20）所述G蛋白的氨基酸取代包含T438S、F453L、T471I和Y487H；
21）所述G蛋白的氨基酸取代包含F453L、T471I和Y487H；
22）所述G蛋白的氨基酸取代包含T471I和Y487H；
23）所述G蛋白的氨基酸取代包含Y487H。
8.根据权利要求1-7中任一项所述的溶瘤病毒，其特征在于：所述G蛋白的氨基酸取代包含V53I、A141V、D172Y、K217E、D232G、V331A、V371E、G436D、T438S、F453L、T471I和Y487H。
9.根据权利要求8任一项所述的溶瘤病毒，其特征在于：所述G蛋白包含如SEQ ID NO17所示的氨基酸序列。
10.根据权利要求1-9中任一项所述的溶瘤病毒，其特征在于：所述溶瘤病毒包含棒状病毒。
11.根据权利要求1-10中任一项所述的溶瘤病毒，其特征在于：所述溶瘤病毒包含水疱性口炎病毒（Vesicular Stomatitis Virus，简称“VSV”）。
12.根据权利要求1-11中任一项所述的溶瘤病毒，其特征在于：所述溶瘤病毒包含VSV病毒Indiana MuddSummer亚型。
13.根据权利要求1-12中任一项所述的溶瘤病毒，其特征在于：所述溶瘤病毒包含或表达外源目的蛋白。
14.根据权利要求1-13中任一项所述的溶瘤病毒，其特征在于：所述溶瘤病毒包含核酸分子，所述核酸分子包含编码所述具有氨基酸取代的M蛋白的核酸序列和编码所述具有氨基酸取代的G蛋白的核酸序列。
15.根据权利要求14所述的溶瘤病毒，其特征在于：所述核酸分子包含编码所述外源目的蛋白的核酸序列。
16.根据权利要求15所述的溶瘤病毒，其特征在于：所述核酸分子中所述编码外源目的蛋白的核酸序列位于所述编码所述具有L蛋白的核酸序列和所述编码所述具有氨基酸取代的G蛋白的核酸序列之间。
17.根据权利要求16所述的溶瘤病毒，其特征在于：所述核酸分子编码所述溶瘤病毒的N蛋白、L蛋白和P蛋白。
18.溶瘤病毒表达载体，其特征在于，所述溶瘤病毒表达载体能够表达权利要求1-17中任一项所述的溶瘤病毒。
19.病毒生产细胞，其特征在于：所述病毒生产细胞能够产生权利要求1-17中任一项所述的溶瘤病毒。
20.药物组合物，其特征在于：所述药物组合物包括权利要求1-17中任一项所述的溶瘤病毒，以及任选地药学上可接受的载剂。
21.权利要求1-17中任一项所述的溶瘤病毒、权利要求18所述的溶瘤病毒表达载体、权利要求19所述的病毒生产细胞和/或权利要求20所述的药物组合物的制备方法。
22.权利要求1-17中任一项所述的溶瘤病毒、权利要求18所述的溶瘤病毒表达载体、权利要求19所述的病毒生产细胞和/或权利要求20所述的药物组合物在制备用于预防和/或治疗疾病和/或病症的药物中的用途。
23.根据权利要求22所述的用途，其特征在于：所述溶瘤病毒、所述溶瘤病毒表达载体、所述病毒生产细胞和/或所述药物组合物用于持续杀伤异常增生性细胞的方法中。
24.根据权利要求23所述的用途，其特征在于：所述异常增生性细胞选自肿瘤细胞或肿瘤组织的相关细胞。
25.根据权利要求24所述的用途，其特征在于：所述肿瘤细胞为癌细胞或转移性癌细胞。
26.根据权利要求22-24任一项所述的用途，其中所述的用途包括：实体瘤和/或血液瘤。
Priority Applications (8)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110798531.XA CN115612674A (zh) | 2021-07-14 | 2021-07-14 | 一种溶瘤病毒及其用途 |
| JP2024501981A JP2024525781A (ja) | 2021-07-14 | 2022-07-08 | 腫瘍溶解性ウイルス及びその使用 |
| KR1020247002721A KR20240026493A (ko) | 2021-07-14 | 2022-07-08 | 종양용해 바이러스 및 그 용도 |
| PCT/CN2022/104525 WO2023284635A1 (zh) | 2021-07-14 | 2022-07-08 | 一种溶瘤病毒及其用途 |
| CA3225820A CA3225820A1 (en) | 2021-07-14 | 2022-07-08 | Oncolytic virus and use thereof |
| AU2022312026A AU2022312026A1 (en) | 2021-07-14 | 2022-07-08 | Oncolytic virus and use thereof |
| EP22841271.4A EP4361257A1 (en) | 2021-07-14 | 2022-07-08 | Oncolytic virus and use thereof |
| US18/412,784 US20240226209A1 (en) | 2021-07-14 | 2024-01-15 | Oncolytic virus and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110798531.XA CN115612674A (zh) | 2021-07-14 | 2021-07-14 | 一种溶瘤病毒及其用途 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115612674A true CN115612674A (zh) | 2023-01-17 |
Family
ID=84854426
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110798531.XA Pending CN115612674A (zh) | 2021-07-14 | 2021-07-14 | 一种溶瘤病毒及其用途 |
Country Status (8)
| Country | Link |
|---|---|
| US (1) | US20240226209A1 (zh) |
| EP (1) | EP4361257A1 (zh) |
| JP (1) | JP2024525781A (zh) |
| KR (1) | KR20240026493A (zh) |
| CN (1) | CN115612674A (zh) |
| AU (1) | AU2022312026A1 (zh) |
| CA (1) | CA3225820A1 (zh) |
| WO (1) | WO2023284635A1 (zh) |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2319424T3 (es) * | 2003-03-27 | 2009-05-07 | Ottawa Health Research Institute | Virus mutantes de la estomatitis vesivular y sus usos. |
| TWI688401B (zh) | 2013-09-13 | 2020-03-21 | 美商安進公司 | 用於治療骨髓性白血病的表觀遺傳因子與靶向ｃｄ３３及ｃｄ３之雙特異性化合物的組合 |
| CN111320677A (zh) * | 2018-12-14 | 2020-06-23 | 苏州奥特铭医药科技有限公司 | 溶瘤病毒疫苗和过继性免疫细胞联合疗法 |
| CN111939262B (zh) * | 2019-05-17 | 2021-09-17 | 睿丰康生物医药科技(浙江)有限公司 | 一种治疗肿瘤或癌症的药物组合物和其应用 |
| CN111467489B (zh) * | 2020-05-12 | 2022-06-03 | 上海荣瑞医药科技有限公司 | 一种治疗肿瘤的药物 |
| CN111286493B (zh) * | 2020-05-12 | 2020-10-27 | 上海荣瑞医药科技有限公司 | 一种溶瘤病毒疫苗及其与免疫细胞联合治疗肿瘤的药物 |
-
2021
- 2021-07-14 CN CN202110798531.XA patent/CN115612674A/zh active Pending
-
2022
- 2022-07-08 WO PCT/CN2022/104525 patent/WO2023284635A1/zh active Application Filing
- 2022-07-08 JP JP2024501981A patent/JP2024525781A/ja active Pending
- 2022-07-08 AU AU2022312026A patent/AU2022312026A1/en active Pending
- 2022-07-08 CA CA3225820A patent/CA3225820A1/en active Pending
- 2022-07-08 KR KR1020247002721A patent/KR20240026493A/ko active Search and Examination
- 2022-07-08 EP EP22841271.4A patent/EP4361257A1/en active Pending
-
2024
- 2024-01-15 US US18/412,784 patent/US20240226209A1/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| KR20240026493A (ko) | 2024-02-28 |
| WO2023284635A1 (zh) | 2023-01-19 |
| US20240226209A1 (en) | 2024-07-11 |
| CA3225820A1 (en) | 2023-01-19 |
| EP4361257A1 (en) | 2024-05-01 |
| JP2024525781A (ja) | 2024-07-12 |
| AU2022312026A1 (en) | 2024-02-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2023143023A1 (zh) | 溶瘤病毒及其应用 | |
| JP7468958B2 (ja) | 腫瘍溶解性ウイルスワクチン及びそれと免疫細胞の併用による腫瘍治療薬物 | |
| CN111467489B (zh) | 一种治疗肿瘤的药物 | |
| CN110996980B (zh) | 一种用于治疗肿瘤的病毒 | |
| JP7336791B2 (ja) | 腫瘍を治療するためのコクサッキーウイルスｂ群 | |
| US11707496B2 (en) | Echovirus for treatment of tumors | |
| CN115612674A (zh) | 一种溶瘤病毒及其用途 | |
| CN112996802A (zh) | 用于制备抗肿瘤沙粒病毒的方法以及沙粒病毒突变体 | |
| WO2019183802A1 (zh) | 一种溶瘤棒状病毒减毒株及其在肿瘤治疗中的应用 | |
| CN113939304B (zh) | M1病毒变异体及其应用 | |
| CN114213505A (zh) | 一种适用于特异感染u87-mg细胞的腺相关病毒突变体 | |
| WO2024012277A1 (zh) | 重组溶瘤病毒及其应用 | |
| CN112538466A (zh) | 一种柯萨奇病毒及其在制备抗肝癌药物中的应用 |
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 |