CN108611395A - 一种抗人乙肝病毒药物的筛选方法 - Google Patents
一种抗人乙肝病毒药物的筛选方法 Download PDFInfo
- Publication number
- CN108611395A CN108611395A CN201611138490.7A CN201611138490A CN108611395A CN 108611395 A CN108611395 A CN 108611395A CN 201611138490 A CN201611138490 A CN 201611138490A CN 108611395 A CN108611395 A CN 108611395A
- Authority
- CN
- China
- Prior art keywords
- peptide fragment
- leu
- seq
- gly
- lys
- 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
- 239000003814 drug Substances 0.000 title claims abstract description 35
- 229940079593 drug Drugs 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 29
- 238000012216 screening Methods 0.000 title claims abstract description 19
- 108010033276 Peptide Fragments Proteins 0.000 claims abstract description 51
- 102000007079 Peptide Fragments Human genes 0.000 claims abstract description 51
- MURGITYSBWUQTI-UHFFFAOYSA-N fluorescin Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC=C(O)C=C2OC2=CC(O)=CC=C21 MURGITYSBWUQTI-UHFFFAOYSA-N 0.000 claims abstract description 34
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 22
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 22
- 108020001507 fusion proteins Proteins 0.000 claims abstract description 13
- 102000037865 fusion proteins Human genes 0.000 claims abstract description 13
- 239000013642 negative control Substances 0.000 claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 8
- 238000005516 engineering process Methods 0.000 claims abstract description 6
- JARGNLJYKBUKSJ-KGZKBUQUSA-N (2r)-2-amino-5-[[(2r)-1-(carboxymethylamino)-3-hydroxy-1-oxopropan-2-yl]amino]-5-oxopentanoic acid;hydrobromide Chemical compound Br.OC(=O)[C@H](N)CCC(=O)N[C@H](CO)C(=O)NCC(O)=O JARGNLJYKBUKSJ-KGZKBUQUSA-N 0.000 claims description 3
- YMHOBZXQZVXHBM-UHFFFAOYSA-N 2,5-dimethoxy-4-bromophenethylamine Chemical compound COC1=CC(CCN)=C(OC)C=C1Br YMHOBZXQZVXHBM-UHFFFAOYSA-N 0.000 claims description 3
- 241000545067 Venus Species 0.000 claims description 3
- 108010044804 gamma-glutamyl-seryl-glycine Proteins 0.000 claims description 3
- 108700026078 glutathione trisulfide Proteins 0.000 claims description 3
- 108010043121 Green Fluorescent Proteins Proteins 0.000 claims description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 claims description 2
- 101000600434 Homo sapiens Putative uncharacterized protein encoded by MIR7-3HG Proteins 0.000 claims description 2
- 102100037401 Putative uncharacterized protein encoded by MIR7-3HG Human genes 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- 239000005090 green fluorescent protein Substances 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 12
- 241001062009 Indigofera Species 0.000 claims 1
- 108090000765 processed proteins & peptides Proteins 0.000 claims 1
- 239000003443 antiviral agent Substances 0.000 abstract description 5
- 230000008569 process Effects 0.000 abstract description 5
- 238000007877 drug screening Methods 0.000 abstract description 3
- 238000012544 monitoring process Methods 0.000 abstract description 2
- 238000011160 research Methods 0.000 abstract description 2
- 238000013537 high throughput screening Methods 0.000 abstract 1
- 150000001413 amino acids Chemical group 0.000 description 24
- RBRNEFJTEHPDSL-ACRUOGEOSA-N Phe-Phe-Lys Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 RBRNEFJTEHPDSL-ACRUOGEOSA-N 0.000 description 12
- 239000000203 mixture Substances 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 11
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 10
- 208000002672 hepatitis B Diseases 0.000 description 10
- LNENWJXDHCFVOF-DCAQKATOSA-N Asp-His-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CC(=O)O)N LNENWJXDHCFVOF-DCAQKATOSA-N 0.000 description 8
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 7
- 108010057821 leucylproline Proteins 0.000 description 7
- XVZCXCTYGHPNEM-IHRRRGAJSA-N (2s)-1-[(2s)-2-[[(2s)-2-amino-4-methylpentanoyl]amino]-4-methylpentanoyl]pyrrolidine-2-carboxylic acid Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(O)=O XVZCXCTYGHPNEM-IHRRRGAJSA-N 0.000 description 6
- YNSUUAOAFCVINY-OSUNSFLBSA-N Arg-Thr-Ile Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YNSUUAOAFCVINY-OSUNSFLBSA-N 0.000 description 6
- QCTOLCVIGRLMQS-HRCADAONSA-N Arg-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O QCTOLCVIGRLMQS-HRCADAONSA-N 0.000 description 6
- MEFGKQUUYZOLHM-GMOBBJLQSA-N Asn-Arg-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MEFGKQUUYZOLHM-GMOBBJLQSA-N 0.000 description 6
- QNNBHTFDFFFHGC-KKUMJFAQSA-N Asn-Tyr-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O QNNBHTFDFFFHGC-KKUMJFAQSA-N 0.000 description 6
- VBVKSAFJPVXMFJ-CIUDSAMLSA-N Asp-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)O)N VBVKSAFJPVXMFJ-CIUDSAMLSA-N 0.000 description 6
- FANQWNCPNFEPGZ-WHFBIAKZSA-N Asp-Asp-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O FANQWNCPNFEPGZ-WHFBIAKZSA-N 0.000 description 6
- WBDWQKRLTVCDSY-WHFBIAKZSA-N Asp-Gly-Asp Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O WBDWQKRLTVCDSY-WHFBIAKZSA-N 0.000 description 6
- NAPULYCVEVVFRB-HEIBUPTGSA-N Cys-Thr-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](N)CS NAPULYCVEVVFRB-HEIBUPTGSA-N 0.000 description 6
- LTXLIIZACMCQTO-GUBZILKMSA-N Gln-His-Asp Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)N)N LTXLIIZACMCQTO-GUBZILKMSA-N 0.000 description 6
- MTAOBYXRYJZRGQ-WDSKDSINSA-N Glu-Gly-Asp Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O MTAOBYXRYJZRGQ-WDSKDSINSA-N 0.000 description 6
- OGNJZUXUTPQVBR-BQBZGAKWSA-N Glu-Gly-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O OGNJZUXUTPQVBR-BQBZGAKWSA-N 0.000 description 6
- IVGJYOOGJLFKQE-AVGNSLFASA-N Glu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N IVGJYOOGJLFKQE-AVGNSLFASA-N 0.000 description 6
- BGVYNAQWHSTTSP-BYULHYEWSA-N Gly-Asn-Ile Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BGVYNAQWHSTTSP-BYULHYEWSA-N 0.000 description 6
- KQDMENMTYNBWMR-WHFBIAKZSA-N Gly-Asp-Ala Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O KQDMENMTYNBWMR-WHFBIAKZSA-N 0.000 description 6
- MHXKHKWHPNETGG-QWRGUYRKSA-N Gly-Lys-Leu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O MHXKHKWHPNETGG-QWRGUYRKSA-N 0.000 description 6
- IGBBXBFSLKRHJB-BZSNNMDCSA-N His-Lys-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CN=CN1 IGBBXBFSLKRHJB-BZSNNMDCSA-N 0.000 description 6
- HGNUKGZQASSBKQ-PCBIJLKTSA-N Ile-Asp-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N HGNUKGZQASSBKQ-PCBIJLKTSA-N 0.000 description 6
- VBZOAGIPCULURB-QWRGUYRKSA-N Leu-Gly-His Chemical compound CC(C)C[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N VBZOAGIPCULURB-QWRGUYRKSA-N 0.000 description 6
- XVZCXCTYGHPNEM-UHFFFAOYSA-N Leu-Leu-Pro Natural products CC(C)CC(N)C(=O)NC(CC(C)C)C(=O)N1CCCC1C(O)=O XVZCXCTYGHPNEM-UHFFFAOYSA-N 0.000 description 6
- QESXLSQLQHHTIX-RHYQMDGZSA-N Leu-Val-Thr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QESXLSQLQHHTIX-RHYQMDGZSA-N 0.000 description 6
- SKRGVGLIRUGANF-AVGNSLFASA-N Lys-Leu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SKRGVGLIRUGANF-AVGNSLFASA-N 0.000 description 6
- OIQSIMFSVLLWBX-VOAKCMCISA-N Lys-Leu-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OIQSIMFSVLLWBX-VOAKCMCISA-N 0.000 description 6
- NMELOOXSGDRBRU-YUMQZZPRSA-N Pro-Glu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CCC(=O)O)NC(=O)[C@@H]1CCCN1 NMELOOXSGDRBRU-YUMQZZPRSA-N 0.000 description 6
- 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 6
- IYCBDVBJWDXQRR-FXQIFTODSA-N Ser-Ala-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O IYCBDVBJWDXQRR-FXQIFTODSA-N 0.000 description 6
- RJHJPZQOMKCSTP-CIUDSAMLSA-N Ser-His-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(O)=O RJHJPZQOMKCSTP-CIUDSAMLSA-N 0.000 description 6
- JGUWRQWULDWNCM-FXQIFTODSA-N Ser-Val-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O JGUWRQWULDWNCM-FXQIFTODSA-N 0.000 description 6
- CAGTXGDOIFXLPC-KZVJFYERSA-N Thr-Arg-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CCCN=C(N)N CAGTXGDOIFXLPC-KZVJFYERSA-N 0.000 description 6
- KZSYAEWQMJEGRZ-RHYQMDGZSA-N Thr-Leu-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O KZSYAEWQMJEGRZ-RHYQMDGZSA-N 0.000 description 6
- PELIQFPESHBTMA-WLTAIBSBSA-N Thr-Tyr-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=C(O)C=C1 PELIQFPESHBTMA-WLTAIBSBSA-N 0.000 description 6
- IKUMWSDCGQVGHC-UMPQAUOISA-N Trp-Pro-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CC2=CNC3=CC=CC=C32)N)O IKUMWSDCGQVGHC-UMPQAUOISA-N 0.000 description 6
- KLOZTPOXVVRVAQ-DZKIICNBSA-N Tyr-Val-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 KLOZTPOXVVRVAQ-DZKIICNBSA-N 0.000 description 6
- UDNYEPLJTRDMEJ-RCOVLWMOSA-N Val-Asn-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)O)N UDNYEPLJTRDMEJ-RCOVLWMOSA-N 0.000 description 6
- ROLGIBMFNMZANA-GVXVVHGQSA-N Val-Glu-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](C(C)C)N ROLGIBMFNMZANA-GVXVVHGQSA-N 0.000 description 6
- HPANGHISDXDUQY-ULQDDVLXSA-N Val-Lys-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N HPANGHISDXDUQY-ULQDDVLXSA-N 0.000 description 6
- GTACFKZDQFTVAI-STECZYCISA-N Val-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC1=CC=C(O)C=C1 GTACFKZDQFTVAI-STECZYCISA-N 0.000 description 6
- 108010005233 alanylglutamic acid Proteins 0.000 description 6
- 108010042598 glutamyl-aspartyl-glycine Proteins 0.000 description 6
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 6
- 108010072405 glycyl-aspartyl-glycine Proteins 0.000 description 6
- 108010038983 glycyl-histidyl-lysine Proteins 0.000 description 6
- 108010015792 glycyllysine Proteins 0.000 description 6
- 108010087823 glycyltyrosine Proteins 0.000 description 6
- 108010003700 lysyl aspartic acid Proteins 0.000 description 6
- 108010012988 lysyl-glutamyl-aspartyl-glycine Proteins 0.000 description 6
- 108010051242 phenylalanylserine Proteins 0.000 description 6
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 6
- 108010061238 threonyl-glycine Proteins 0.000 description 6
- 108700004896 tripeptide FEG Proteins 0.000 description 6
- 108091005957 yellow fluorescent proteins Proteins 0.000 description 6
- LZRNYBIJOSKKRJ-XVYDVKMFSA-N Ala-Asp-His Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N LZRNYBIJOSKKRJ-XVYDVKMFSA-N 0.000 description 5
- 102100038567 Properdin Human genes 0.000 description 5
- QUILOGWWLXMSAT-IHRRRGAJSA-N Tyr-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O QUILOGWWLXMSAT-IHRRRGAJSA-N 0.000 description 5
- 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 4
- SHYYAQLDNVHPFT-DLOVCJGASA-N Ala-Asn-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 SHYYAQLDNVHPFT-DLOVCJGASA-N 0.000 description 4
- RTFXPCYMDYBZNQ-SRVKXCTJSA-N Asn-Tyr-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O RTFXPCYMDYBZNQ-SRVKXCTJSA-N 0.000 description 4
- UIKLEGZPIOXFHJ-DLOVCJGASA-N Cys-Phe-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(O)=O UIKLEGZPIOXFHJ-DLOVCJGASA-N 0.000 description 4
- ITZOBNKQDZEOCE-NHCYSSNCSA-N Gly-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)CN ITZOBNKQDZEOCE-NHCYSSNCSA-N 0.000 description 4
- TWTPDFFBLQEBOE-IUCAKERBSA-N Gly-Leu-Gln Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O TWTPDFFBLQEBOE-IUCAKERBSA-N 0.000 description 4
- KBDIBHQICWDGDL-PPCPHDFISA-N Ile-Thr-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N KBDIBHQICWDGDL-PPCPHDFISA-N 0.000 description 4
- QNBVTHNJGCOVFA-AVGNSLFASA-N Leu-Leu-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCC(O)=O QNBVTHNJGCOVFA-AVGNSLFASA-N 0.000 description 4
- QNTJIDXQHWUBKC-BZSNNMDCSA-N Leu-Lys-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QNTJIDXQHWUBKC-BZSNNMDCSA-N 0.000 description 4
- MXMDJEJWERYPMO-XUXIUFHCSA-N Lys-Ile-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MXMDJEJWERYPMO-XUXIUFHCSA-N 0.000 description 4
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 4
- MWQXFDIQXIXPMS-UNQGMJICSA-N Phe-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC1=CC=CC=C1)N)O MWQXFDIQXIXPMS-UNQGMJICSA-N 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 108010050848 glycylleucine Proteins 0.000 description 4
- LSLIRHLIUDVNBN-CIUDSAMLSA-N Ala-Asp-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN LSLIRHLIUDVNBN-CIUDSAMLSA-N 0.000 description 3
- WUQXMTITJLFXAU-JIOCBJNQSA-N Asn-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)N)N)O WUQXMTITJLFXAU-JIOCBJNQSA-N 0.000 description 3
- POTCZYQVVNXUIG-BQBZGAKWSA-N Asp-Gly-Pro Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O POTCZYQVVNXUIG-BQBZGAKWSA-N 0.000 description 3
- BWJZSLQJNBSUPM-FXQIFTODSA-N Asp-Pro-Asn Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O BWJZSLQJNBSUPM-FXQIFTODSA-N 0.000 description 3
- 101710132601 Capsid protein Proteins 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- JNENSVNAUWONEZ-GUBZILKMSA-N Gln-Lys-Asn Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O JNENSVNAUWONEZ-GUBZILKMSA-N 0.000 description 3
- DSPQRJXOIXHOHK-WDSKDSINSA-N Glu-Asp-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O DSPQRJXOIXHOHK-WDSKDSINSA-N 0.000 description 3
- SJJHXJDSNQJMMW-SRVKXCTJSA-N Glu-Lys-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O SJJHXJDSNQJMMW-SRVKXCTJSA-N 0.000 description 3
- OJNZVYSGVYLQIN-BQBZGAKWSA-N Gly-Met-Asp Chemical compound [H]NCC(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(O)=O OJNZVYSGVYLQIN-BQBZGAKWSA-N 0.000 description 3
- 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 3
- NGRPGJGKJMUGDM-XVKPBYJWSA-N Gly-Val-Gln Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O NGRPGJGKJMUGDM-XVKPBYJWSA-N 0.000 description 3
- UZZXGLOJRZKYEL-DJFWLOJKSA-N His-Asn-Ile Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UZZXGLOJRZKYEL-DJFWLOJKSA-N 0.000 description 3
- KFVUBLZRFSVDGO-BYULHYEWSA-N Ile-Gly-Asp Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(O)=O KFVUBLZRFSVDGO-BYULHYEWSA-N 0.000 description 3
- 102100034349 Integrase Human genes 0.000 description 3
- AMSSKPUHBUQBOQ-SRVKXCTJSA-N Leu-Ser-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N AMSSKPUHBUQBOQ-SRVKXCTJSA-N 0.000 description 3
- QQUJSUFWEDZQQY-AVGNSLFASA-N Lys-Gln-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CCCCN QQUJSUFWEDZQQY-AVGNSLFASA-N 0.000 description 3
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 3
- NSGZILIDHCIZAM-KKUMJFAQSA-N Tyr-Leu-Ser Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N NSGZILIDHCIZAM-KKUMJFAQSA-N 0.000 description 3
- 108010092854 aspartyllysine Proteins 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 108010053725 prolylvaline Proteins 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 108010051110 tyrosyl-lysine Proteins 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- LMFXXZPPZDCPTA-ZKWXMUAHSA-N Ala-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N LMFXXZPPZDCPTA-ZKWXMUAHSA-N 0.000 description 2
- XVBDDUPJVQXDSI-PEFMBERDSA-N Asn-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N XVBDDUPJVQXDSI-PEFMBERDSA-N 0.000 description 2
- BEHQTVDBCLSCBY-CFMVVWHZSA-N Asn-Tyr-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BEHQTVDBCLSCBY-CFMVVWHZSA-N 0.000 description 2
- PDECQIHABNQRHN-GUBZILKMSA-N Asp-Glu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(O)=O PDECQIHABNQRHN-GUBZILKMSA-N 0.000 description 2
- 108091036055 CccDNA Proteins 0.000 description 2
- CHRCKSPMGYDLIA-SRVKXCTJSA-N Cys-Phe-Ser Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O CHRCKSPMGYDLIA-SRVKXCTJSA-N 0.000 description 2
- KUBFPYIMAGXGBT-ACZMJKKPSA-N Gln-Ser-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O KUBFPYIMAGXGBT-ACZMJKKPSA-N 0.000 description 2
- LUJVWKKYHSLULQ-ZKWXMUAHSA-N Gly-Ile-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)CN LUJVWKKYHSLULQ-ZKWXMUAHSA-N 0.000 description 2
- 241000700721 Hepatitis B virus Species 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- LAGPXKYZCCTSGQ-JYJNAYRXSA-N Leu-Glu-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LAGPXKYZCCTSGQ-JYJNAYRXSA-N 0.000 description 2
- UCDHVOALNXENLC-KBPBESRZSA-N Leu-Gly-Tyr Chemical compound CC(C)C[C@H]([NH3+])C(=O)NCC(=O)N[C@H](C([O-])=O)CC1=CC=C(O)C=C1 UCDHVOALNXENLC-KBPBESRZSA-N 0.000 description 2
- BGZCJDGBBUUBHA-KKUMJFAQSA-N Leu-Lys-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O BGZCJDGBBUUBHA-KKUMJFAQSA-N 0.000 description 2
- RNYLNYTYMXACRI-VFAJRCTISA-N Leu-Thr-Trp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O RNYLNYTYMXACRI-VFAJRCTISA-N 0.000 description 2
- GQUDMNDPQTXZRV-DCAQKATOSA-N Lys-Arg-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O GQUDMNDPQTXZRV-DCAQKATOSA-N 0.000 description 2
- MDDUIRLQCYVRDO-NHCYSSNCSA-N Lys-Val-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN MDDUIRLQCYVRDO-NHCYSSNCSA-N 0.000 description 2
- QAVZUKIPOMBLMC-AVGNSLFASA-N Met-Val-Leu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC(C)C QAVZUKIPOMBLMC-AVGNSLFASA-N 0.000 description 2
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 2
- WFHRXJOZEXUKLV-IRXDYDNUSA-N Phe-Gly-Tyr Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 WFHRXJOZEXUKLV-IRXDYDNUSA-N 0.000 description 2
- ZUQACJLOHYRVPJ-DKIMLUQUSA-N Phe-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=CC=C1 ZUQACJLOHYRVPJ-DKIMLUQUSA-N 0.000 description 2
- OBVCYFIHIIYIQF-CIUDSAMLSA-N Pro-Asn-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O OBVCYFIHIIYIQF-CIUDSAMLSA-N 0.000 description 2
- PPNPDKGQRFSCAC-CIUDSAMLSA-N Ser-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPNPDKGQRFSCAC-CIUDSAMLSA-N 0.000 description 2
- LLSLRQOEAFCZLW-NRPADANISA-N Ser-Val-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O LLSLRQOEAFCZLW-NRPADANISA-N 0.000 description 2
- NJEMRSFGDNECGF-GCJQMDKQSA-N Thr-Ala-Asp Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O NJEMRSFGDNECGF-GCJQMDKQSA-N 0.000 description 2
- 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 2
- VTMGKRABARCZAX-OSUNSFLBSA-N Thr-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)[C@@H](C)O VTMGKRABARCZAX-OSUNSFLBSA-N 0.000 description 2
- VFOHXOLPLACADK-GVXVVHGQSA-N Val-Gln-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](C(C)C)N VFOHXOLPLACADK-GVXVVHGQSA-N 0.000 description 2
- 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 2
- 108010013835 arginine glutamate Proteins 0.000 description 2
- 108010068380 arginylarginine Proteins 0.000 description 2
- 108010038633 aspartylglutamate Proteins 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 108010045126 glycyl-tyrosyl-glycine Proteins 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 230000005571 horizontal transmission Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 210000005229 liver cell Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002777 nucleoside Substances 0.000 description 2
- 150000003833 nucleoside derivatives Chemical class 0.000 description 2
- 230000005180 public health Effects 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 2
- 108010038745 tryptophylglycine Proteins 0.000 description 2
- 230000005570 vertical transmission Effects 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 206010059193 Acute hepatitis B Diseases 0.000 description 1
- 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 1
- IETUUAHKCHOQHP-KZVJFYERSA-N Ala-Thr-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@H](C)N)[C@@H](C)O)C(O)=O IETUUAHKCHOQHP-KZVJFYERSA-N 0.000 description 1
- XPSGESXVBSQZPL-SRVKXCTJSA-N Arg-Arg-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O XPSGESXVBSQZPL-SRVKXCTJSA-N 0.000 description 1
- UISQLSIBJKEJSS-GUBZILKMSA-N Arg-Arg-Ser Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(O)=O UISQLSIBJKEJSS-GUBZILKMSA-N 0.000 description 1
- NABSCJGZKWSNHX-RCWTZXSCSA-N Arg-Arg-Thr Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H]([C@H](O)C)C(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N NABSCJGZKWSNHX-RCWTZXSCSA-N 0.000 description 1
- YFBGNGASPGRWEM-DCAQKATOSA-N Arg-Asp-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N YFBGNGASPGRWEM-DCAQKATOSA-N 0.000 description 1
- FEZJJKXNPSEYEV-CIUDSAMLSA-N Arg-Gln-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O FEZJJKXNPSEYEV-CIUDSAMLSA-N 0.000 description 1
- HQIZDMIGUJOSNI-IUCAKERBSA-N Arg-Gly-Arg Chemical compound N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O HQIZDMIGUJOSNI-IUCAKERBSA-N 0.000 description 1
- QEHMMRSQJMOYNO-DCAQKATOSA-N Arg-His-Asn Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N QEHMMRSQJMOYNO-DCAQKATOSA-N 0.000 description 1
- NUHQMYUWLUSRJX-BIIVOSGPSA-N Asn-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)N)N NUHQMYUWLUSRJX-BIIVOSGPSA-N 0.000 description 1
- OLGCWMNDJTWQAG-GUBZILKMSA-N Asn-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(N)=O OLGCWMNDJTWQAG-GUBZILKMSA-N 0.000 description 1
- HDHZCEDPLTVHFZ-GUBZILKMSA-N Asn-Leu-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O HDHZCEDPLTVHFZ-GUBZILKMSA-N 0.000 description 1
- OMMIEVATLAGRCK-BYPYZUCNSA-N Asp-Gly-Gly Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)NCC(O)=O OMMIEVATLAGRCK-BYPYZUCNSA-N 0.000 description 1
- SNDBKTFJWVEVPO-WHFBIAKZSA-N Asp-Gly-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SNDBKTFJWVEVPO-WHFBIAKZSA-N 0.000 description 1
- SWTQDYFZVOJVLL-KKUMJFAQSA-N Asp-His-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC(=O)O)N)O SWTQDYFZVOJVLL-KKUMJFAQSA-N 0.000 description 1
- KPSHWSWFPUDEGF-FXQIFTODSA-N Asp-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC(O)=O KPSHWSWFPUDEGF-FXQIFTODSA-N 0.000 description 1
- XZKJEOMFLDVXJG-KATARQTJSA-N Cys-Leu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)N)O XZKJEOMFLDVXJG-KATARQTJSA-N 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- CITDWMLWXNUQKD-FXQIFTODSA-N Gln-Gln-Asn Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N CITDWMLWXNUQKD-FXQIFTODSA-N 0.000 description 1
- HYPVLWGNBIYTNA-GUBZILKMSA-N Gln-Leu-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O HYPVLWGNBIYTNA-GUBZILKMSA-N 0.000 description 1
- NYCVMJGIJYQWDO-CIUDSAMLSA-N Gln-Ser-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NYCVMJGIJYQWDO-CIUDSAMLSA-N 0.000 description 1
- KPNWAJMEMRCLAL-GUBZILKMSA-N Gln-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N KPNWAJMEMRCLAL-GUBZILKMSA-N 0.000 description 1
- ITYRYNUZHPNCIK-GUBZILKMSA-N Glu-Ala-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O ITYRYNUZHPNCIK-GUBZILKMSA-N 0.000 description 1
- COSBSYQVPSODFX-GUBZILKMSA-N Glu-His-Cys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCC(=O)O)N COSBSYQVPSODFX-GUBZILKMSA-N 0.000 description 1
- MWMJCGBSIORNCD-AVGNSLFASA-N Glu-Leu-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O MWMJCGBSIORNCD-AVGNSLFASA-N 0.000 description 1
- YRMZCZIRHYCNHX-RYUDHWBXSA-N Glu-Phe-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(O)=O YRMZCZIRHYCNHX-RYUDHWBXSA-N 0.000 description 1
- DAHLWSFUXOHMIA-FXQIFTODSA-N Glu-Ser-Gln Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O DAHLWSFUXOHMIA-FXQIFTODSA-N 0.000 description 1
- HMJULNMJWOZNFI-XHNCKOQMSA-N Glu-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)O)N)C(=O)O HMJULNMJWOZNFI-XHNCKOQMSA-N 0.000 description 1
- LWYUQLZOIORFFJ-XKBZYTNZSA-N Glu-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O LWYUQLZOIORFFJ-XKBZYTNZSA-N 0.000 description 1
- DLISPGXMKZTWQG-IFFSRLJSSA-N Glu-Thr-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O DLISPGXMKZTWQG-IFFSRLJSSA-N 0.000 description 1
- LLZXNUUIBOALNY-QWRGUYRKSA-N Gly-Leu-Lys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN LLZXNUUIBOALNY-QWRGUYRKSA-N 0.000 description 1
- ZLCLYFGMKFCDCN-XPUUQOCRSA-N Gly-Ser-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CO)NC(=O)CN)C(O)=O ZLCLYFGMKFCDCN-XPUUQOCRSA-N 0.000 description 1
- COZMNNJEGNPDED-HOCLYGCPSA-N Gly-Val-Trp Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O COZMNNJEGNPDED-HOCLYGCPSA-N 0.000 description 1
- ORERHHPZDDEMSC-VGDYDELISA-N His-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N ORERHHPZDDEMSC-VGDYDELISA-N 0.000 description 1
- WYSJPCTWSBJFCO-AVGNSLFASA-N His-Met-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CC1=CN=CN1)N WYSJPCTWSBJFCO-AVGNSLFASA-N 0.000 description 1
- 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 1
- QTUSJASXLGLJSR-OSUNSFLBSA-N Ile-Arg-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N QTUSJASXLGLJSR-OSUNSFLBSA-N 0.000 description 1
- NPROWIBAWYMPAZ-GUDRVLHUSA-N Ile-Asp-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N NPROWIBAWYMPAZ-GUDRVLHUSA-N 0.000 description 1
- KIMHKBDJQQYLHU-PEFMBERDSA-N Ile-Glu-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N KIMHKBDJQQYLHU-PEFMBERDSA-N 0.000 description 1
- JXMSHKFPDIUYGS-SIUGBPQLSA-N Ile-Glu-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N JXMSHKFPDIUYGS-SIUGBPQLSA-N 0.000 description 1
- NUKXXNFEUZGPRO-BJDJZHNGSA-N Ile-Leu-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)O)N NUKXXNFEUZGPRO-BJDJZHNGSA-N 0.000 description 1
- GVKKVHNRTUFCCE-BJDJZHNGSA-N Ile-Leu-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)O)N GVKKVHNRTUFCCE-BJDJZHNGSA-N 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- 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 1
- FOBUGKUBUJOWAD-IHPCNDPISA-N Leu-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 FOBUGKUBUJOWAD-IHPCNDPISA-N 0.000 description 1
- JVTYXRRFZCEPPK-RHYQMDGZSA-N Leu-Met-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)N)O JVTYXRRFZCEPPK-RHYQMDGZSA-N 0.000 description 1
- SQUFDMCWMFOEBA-KKUMJFAQSA-N Leu-Ser-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 SQUFDMCWMFOEBA-KKUMJFAQSA-N 0.000 description 1
- BTEMNFBEAAOGBR-BZSNNMDCSA-N Leu-Tyr-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BTEMNFBEAAOGBR-BZSNNMDCSA-N 0.000 description 1
- YEIYAQQKADPIBJ-GARJFASQSA-N Lys-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCCCN)N)C(=O)O YEIYAQQKADPIBJ-GARJFASQSA-N 0.000 description 1
- VHGIWFGJIHTASW-FXQIFTODSA-N Met-Ala-Asp Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O VHGIWFGJIHTASW-FXQIFTODSA-N 0.000 description 1
- GODBLDDYHFTUAH-CIUDSAMLSA-N Met-Asp-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCC(O)=O GODBLDDYHFTUAH-CIUDSAMLSA-N 0.000 description 1
- VYXIKLFLGRTANT-HRCADAONSA-N Met-Tyr-Pro Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N2CCC[C@@H]2C(=O)O)N VYXIKLFLGRTANT-HRCADAONSA-N 0.000 description 1
- 108010079364 N-glycylalanine Proteins 0.000 description 1
- 241001597008 Nomeidae Species 0.000 description 1
- XWBJLKDCHJVKAK-KKUMJFAQSA-N Phe-Arg-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N XWBJLKDCHJVKAK-KKUMJFAQSA-N 0.000 description 1
- RFEXGCASCQGGHZ-STQMWFEESA-N Phe-Gly-Arg Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O RFEXGCASCQGGHZ-STQMWFEESA-N 0.000 description 1
- LNLNHXIQPGKRJQ-SRVKXCTJSA-N Pro-Arg-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H]1CCCN1 LNLNHXIQPGKRJQ-SRVKXCTJSA-N 0.000 description 1
- XYHMFGGWNOFUOU-QXEWZRGKSA-N Pro-Ile-Gly Chemical compound OC(=O)CNC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]1CCCN1 XYHMFGGWNOFUOU-QXEWZRGKSA-N 0.000 description 1
- 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 1
- GMJDSFYVTAMIBF-FXQIFTODSA-N Pro-Ser-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O GMJDSFYVTAMIBF-FXQIFTODSA-N 0.000 description 1
- SNGZLPOXVRTNMB-LPEHRKFASA-N Pro-Ser-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CO)C(=O)N2CCC[C@@H]2C(=O)O SNGZLPOXVRTNMB-LPEHRKFASA-N 0.000 description 1
- 208000034189 Sclerosis Diseases 0.000 description 1
- 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 1
- QEDMOZUJTGEIBF-FXQIFTODSA-N Ser-Arg-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O QEDMOZUJTGEIBF-FXQIFTODSA-N 0.000 description 1
- FMDHKPRACUXATF-ACZMJKKPSA-N Ser-Gln-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O FMDHKPRACUXATF-ACZMJKKPSA-N 0.000 description 1
- XUDRHBPSPAPDJP-SRVKXCTJSA-N Ser-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CO XUDRHBPSPAPDJP-SRVKXCTJSA-N 0.000 description 1
- UPLYXVPQLJVWMM-KKUMJFAQSA-N Ser-Phe-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O UPLYXVPQLJVWMM-KKUMJFAQSA-N 0.000 description 1
- NUEHQDHDLDXCRU-GUBZILKMSA-N Ser-Pro-Arg Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O NUEHQDHDLDXCRU-GUBZILKMSA-N 0.000 description 1
- QPPYAWVLAVXISR-DCAQKATOSA-N Ser-Pro-His Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CO)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O QPPYAWVLAVXISR-DCAQKATOSA-N 0.000 description 1
- SZRNDHWMVSFPSP-XKBZYTNZSA-N Ser-Thr-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CO)N)O SZRNDHWMVSFPSP-XKBZYTNZSA-N 0.000 description 1
- VEVYMLNYMULSMS-AVGNSLFASA-N Ser-Tyr-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O VEVYMLNYMULSMS-AVGNSLFASA-N 0.000 description 1
- QNJZOAHSYPXTAB-VEVYYDQMSA-N Thr-Asn-Met Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O QNJZOAHSYPXTAB-VEVYYDQMSA-N 0.000 description 1
- NCXVJIQMWSGRHY-KXNHARMFSA-N Thr-Leu-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N)O NCXVJIQMWSGRHY-KXNHARMFSA-N 0.000 description 1
- DZIKVMCFXIIETR-JSGCOSHPSA-N Trp-Gly-Glu Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O DZIKVMCFXIIETR-JSGCOSHPSA-N 0.000 description 1
- BURPTJBFWIOHEY-UWJYBYFXSA-N Tyr-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 BURPTJBFWIOHEY-UWJYBYFXSA-N 0.000 description 1
- CRWOSTCODDFEKZ-HRCADAONSA-N Tyr-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)C(=O)O CRWOSTCODDFEKZ-HRCADAONSA-N 0.000 description 1
- HZZKQZDUIKVFDZ-AVGNSLFASA-N Tyr-Gln-Ser Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N)O HZZKQZDUIKVFDZ-AVGNSLFASA-N 0.000 description 1
- AEOFMCAKYIQQFY-YDHLFZDLSA-N Tyr-Val-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AEOFMCAKYIQQFY-YDHLFZDLSA-N 0.000 description 1
- XXROXFHCMVXETG-UWVGGRQHSA-N Val-Gly-Val Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O XXROXFHCMVXETG-UWVGGRQHSA-N 0.000 description 1
- QZKVWWIUSQGWMY-IHRRRGAJSA-N Val-Ser-Phe Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 QZKVWWIUSQGWMY-IHRRRGAJSA-N 0.000 description 1
- RTJPAGFXOWEBAI-SRVKXCTJSA-N Val-Val-Arg Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N RTJPAGFXOWEBAI-SRVKXCTJSA-N 0.000 description 1
- LLJLBRRXKZTTRD-GUBZILKMSA-N Val-Val-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N LLJLBRRXKZTTRD-GUBZILKMSA-N 0.000 description 1
- 108010003533 Viral Envelope Proteins Proteins 0.000 description 1
- 208000037628 acute hepatitis B virus infection Diseases 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 208000016350 chronic hepatitis B virus infection Diseases 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000007771 core particle Substances 0.000 description 1
- 108010082025 cyan fluorescent protein Proteins 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000002532 enzyme inhibitor Substances 0.000 description 1
- 229940125532 enzyme inhibitor Drugs 0.000 description 1
- 108091006047 fluorescent proteins Proteins 0.000 description 1
- 102000034287 fluorescent proteins Human genes 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 108010013768 glutamyl-aspartyl-proline Proteins 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- SPSXSWRZQFPVTJ-ZQQKUFEYSA-N hepatitis b vaccine Chemical compound C([C@H](NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](CC1N=CN=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)OC(=O)CNC(=O)CNC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@@H](N)CCCNC(N)=N)C1=CC=CC=C1 SPSXSWRZQFPVTJ-ZQQKUFEYSA-N 0.000 description 1
- 229940124736 hepatitis-B vaccine Drugs 0.000 description 1
- 108010028295 histidylhistidine Proteins 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 108020004084 membrane receptors Proteins 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 108010005942 methionylglycine Proteins 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108010093296 prolyl-prolyl-alanine Proteins 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- 108010004914 prolylarginine Proteins 0.000 description 1
- 108010015796 prolylisoleucine Proteins 0.000 description 1
- 230000001177 retroviral effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 239000003419 rna directed dna polymerase inhibitor Substances 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
- G01N2500/10—Screening for compounds of potential therapeutic value involving cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Hematology (AREA)
- General Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Toxicology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一种抗人乙肝病毒药物的筛选方法。具体为将SEQ ID NO：1所示的HBc分别与荧光蛋白的肽段a和肽段b相连，构建得融合蛋白A和B，所述肽段a和肽段b连接共同组成荧光蛋白的全序列；将融合蛋白A和B通过克隆技术共转化同一受体细胞，用待检测药物处理上述转化的受体细胞，同时以未用药物处理的上述转化的受体细胞作为阴性对照；观察细胞中荧光蛋白的信号，如果无荧光蛋白信号或荧光信号低于阴性对照，说明待检测药物可以抗人乙肝病毒，如果荧光信号与阴性对照信号强度相当，说明待检测药物不可以抗人乙肝病毒。本研究通过设计并构建HBc与荧光蛋白的融合蛋白，利用荧光蛋白信号监测HBc组装的过程，简化药物筛选过程中的读出操作，提供了一种用于HBV抗病毒药物的高通量筛选方法。
Description
技术领域
本发明涉及抗病毒药物筛选技术领域，具体地，涉及一种抗人乙肝病毒药物的筛选方法。
背景技术
人乙型肝炎病毒感染是世界范围内的重要公共健康问题。急性乙肝病毒感染后，仍有8％左右发展为慢性乙肝感染，持续性HBV感染将导致肝硬化，甚至肝癌。我国是乙肝大国，乙肝病毒携带者接近1.3亿人，约占总人口的9％。尽管随着乙肝疫苗的大范围普及，新乙肝感染率得到有效控制，但乙肝携带人口基数大，防治乙肝成为我国公共健康问题的重中之重。乙肝传播途径主要通过垂直传播与水平传播。垂直传播是指母婴传播；水平传播主要通过血液传播。
尽管目前市场上有很多抗HBV药物，主要通过使用干扰素或者核苷类似物进行抗病毒治疗。其中核苷类似物通过抑制HBV复制过程中的逆转录酶活性从而抑制HBV产生。逆转录酶抑制剂虽然可以使患者控制乙肝病毒水平，但是随之产生的耐药性、巨额的医药费用、药物严重的副作用等问题也不容小觑。由于HBV逆转录酶体外表达并不产生酶活性，HBV逆转录酶抑制剂的母环结构主要通过对人免疫缺陷病毒(HIV)的逆转录酶化合物改造修饰。针对HBV本身进行从头抗病毒药物研究成为一种迫切需求。想要获得从头抗病毒药物，必须先建立一种合适的药物筛选方法。
发明内容
本发明的目的是为了克服现有技术的上述不足，提供一种抗人乙肝病毒药物的筛选方法。该方法筛选的药物能抑制HBV核心蛋白(HBc)的组装，可以从根本上达到抑制乙肝病毒的作用。
为了实现上述目的，本发明是通过以下技术方案予以实现的：
一种抗人乙肝病毒药物的筛选方法，包括如下步骤：将SEQ ID NO：1所示的HBc分别与荧光蛋白的肽段a和肽段b相连，构建得融合蛋白A和B，所述肽段a和肽段b连接共同组成荧光蛋白的全序列；将融合蛋白A和B通过克隆技术共转化同一受体细胞，用待检测药物处理上述转化的受体细胞，同时以未用药物处理的上述转化的受体细胞作为阴性对照；观察细胞中荧光蛋白的信号，如果无荧光蛋白信号或荧光信号低于阴性对照，说明待检测药物可以抗人乙肝病毒，如果荧光信号与阴性对照信号强度相当，说明待检测药物不可以抗人乙肝病毒。
在HBV病毒生活周期中，病毒包膜蛋白与肝脏细胞膜受体结合，病毒包膜与宿主细胞膜融合，病毒核心颗粒进入胞内并介导病毒基因组进入胞核，通过利用人肝脏细胞内聚合酶将dsDNA基因组补全并转化为cccDNA，cccDNA进一步转录出3.5kb，2.4kb，2.1kb及0.8kb的mRNA，以前基因组RNA(pgRNA)3.5kb为模板逆转录出基因组DNA并作为编码病毒核心蛋白(HBc)和聚合酶蛋白的模板；HBc多聚化包装乙肝DNA基因组形成包装核心颗粒；乙肝表面抗原HBsAg合成后在粗面内质网中多聚化，并转运至高尔基体前腔以包装核心颗粒，装配好的HBV颗粒与亚病毒颗粒转运至高尔基体进行HBsAg的糖基化修饰，最后分泌出宿主细胞而完成生活周期。病毒核心(HBc)颗粒的组装是HBV复制产生过程不可缺少的一步，如果HBc的组装受到抑制，HBV的基因组将不能完成包装从而阻断HBV的复制(见图1)。如何抑制HBc的组装，将是研发抗HBV药物的一个重要靶点。
优选地，所述荧光蛋白为黄色荧光蛋白(yellow fluorescent protein，YFP)，蓝绿色荧光蛋白(cyan fluorescent protein，CFP)或Venus荧光蛋白(Venus fluorescentprotein,VFP)。
更优选地，所述肽段a的氨基酸序列如SEQ IDNO：2所示，肽段b的氨基酸序列如SEQIDNO：3所示；或，肽段a的氨基酸序列如SEQ ID NO：4所示，肽段b的氨基酸序列如SEQ IDNO：5所示；或，肽段a的氨基酸序列如SEQ ID NO：6所示，肽段b的氨基酸序列如SEQ ID NO：7所示；或，肽段a的氨基酸序列如SEQ ID NO：8所示，肽段b的氨基酸序列如SEQ ID NO：9所示；或，肽段a的氨基酸序列如SEQ ID NO：10所示，肽段b的氨基酸序列如SEQ ID NO：11所示；或，肽段a的氨基酸序列如SEQ ID NO：12所示，肽段b的氨基酸序列如SEQ ID NO：13所示。
SEQ ID NO：1所示的HBc分别与荧光蛋白的肽段a和肽段b相连时，使用linker序列连接，所述Linker序列为(GGSG)n；(GGSGG)n；(GSGSG)n；(GSGGG)n；(GGGGS)n；(GGGSG)n；(GSSSG)n；RPACKIPNDLKQKVMNH；RSIAT。
所述受体细胞为HEK293T细胞、HepG2或Huh7。
与现有技术相比，本发明具有如下有益效果：
本发明通过设计并构建HBc与荧光蛋白的融合蛋白，利用荧光蛋白信号监测HBc组装的过程，简化药物筛选过程中的读出操作，提供了一种用于HBV抗病毒药物的高通量筛选方法。
附图说明
图1为HBV病毒生活周期中核心蛋白的组装流程图。
图2为实施例1各处理的荧光信号检测结果。
图3为表1化合物的荧光信号检测结果。
具体实施方式
下面结合说明书附图和具体实施例对本发明作出进一步地详细阐述，所述实施例只用于解释本发明，并非用于限定本发明的范围。下述实施例中所使用的试验方法如无特殊说明，均为常规方法；所使用的材料、试剂等，如无特殊说明，为可从商业途径得到的试剂和材料。
实施例1
一种抗人乙肝病毒药物的筛选方法，包括如下步骤：
(1)将SEQ ID NO：1所示的HBc通过Linker序列分别与VFP-N1、VFP-C1相连，得融合蛋白HBc-VFP-N1、HBc-VFP-C1，所述VFP-N1为VFP蛋白1-154位氨基酸组成的肽段，VFP-C1为VFP蛋白155-239位氨基酸组成的肽段。其中，Linker序列包括但不限于(GGSG)n；(GGSGG)n；(GSGSG)n；(GSGGG)n；(GGGGS)n；(GGGSG)n；(GSSSG)n；RPACKIPNDLKQKVMNH；RSIAT等。
(2)在6孔板的HEK293T细胞中，通过克隆技术共转染HBc-VFP-N1 0.5ug和HBc-VFP-C1 0.5ug，或共转染HBc-VFP-N1 0.5ug和VFP-C1 0.5ug，或共转染VFP-N1 0.5ug和HBc-VFP-C1 0.5ug，或共转染VFP-N1 0.5ng和VFP-C1 0.5ng。转染48h后观察VFP的表达(Ex488nm/Em 512nm)结果见图2，该实验说明了，只有当HBc-VFP-1N和HBc-VFP-1C共同存在时，才会产生强荧光信号。HBc的组装使得VFP-N1与VFP-C1互相接近产生完整的VFP蛋白结构，进而产生荧光信号。
(3)基于上述研究，在6孔板的HEK293T细胞中，通过克隆技术共转染HBc-VFP-N10.5ug和HBc-VFP-C1 0.5ug，然后用待检测药物处理上述转化的受体细胞，同时以未用药物处理的上述转化的受体细胞作为阴性对照；观察细胞中荧光蛋白的信号，如果无荧光蛋白信号或荧光信号低于阴性对照，说明待检测药物可以抗人乙肝病毒，如果荧光信号与阴性对照信号强度相当，说明待检测药物不可以抗人乙肝病毒。
实施例2
一种抗人乙肝病毒药物的筛选方法，基本步骤同实施例1，不同之处在于，将SEQID NO：1所示的HBc通过Linker序列分别与VFP-N2、VFP-C2相连，得融合蛋白HBc-VFP-N2、HBc-VFP-C2，所述VFP-N2为VFP蛋白1-172位氨基酸组成的肽段，VFP-C1为VFP蛋白173-239位氨基酸组成的肽段。
实施例3
一种抗人乙肝病毒药物的筛选方法，基本步骤同实施例1，不同之处在于，将SEQID NO：1所示的HBc通过Linker序列分别与YFP-N1、YFP-C1相连，得融合蛋白HBc-YFP-N1、HBc-YFP-C1，所述YFP-N1为YFP蛋白1-154位氨基酸组成的肽段，YFP-C1为YFP蛋白155-240位氨基酸组成的肽段。
实施例4
一种抗人乙肝病毒药物的筛选方法，基本步骤同实施例1，不同之处在于，将SEQID NO：1所示的HBc通过Linker序列分别与YFP-N2、YFP-C2相连，得融合蛋白HBc-YFP-N2、HBc-YFP-C2，所述YFP-N2为YFP蛋白1-172位氨基酸组成的肽段，YFP-C2为YFP蛋白173-240位氨基酸组成的肽段。
实施例5
一种抗人乙肝病毒药物的筛选方法，基本步骤同实施例1，不同之处在于，将SEQID NO：1所示的HBc通过Linker序列分别与CFP-N1、CFP-C1相连，得融合蛋白HBc-CFP-N1、HBc-CFP-C1，所述CFP-N1为CFP蛋白1-154位氨基酸组成的肽段，CFP-C1为CFP蛋白155-239位氨基酸组成的肽段。
实施例6
一种抗人乙肝病毒药物的筛选方法，基本步骤同实施例1，不同之处在于，将SEQID NO：1所示的HBc通过Linker序列分别与CFP-N2、CFP-C2相连，得融合蛋白HBc-CFP-N2、HBc-CFP-C2，所述CFP-N2为CFP蛋白1-172位氨基酸组成的肽段，CFP-C1为CFP蛋白173-239位氨基酸组成的肽段。
应用例1
使用本发明的方法检测表1中的化合物是否为抗乙肝病毒药物，结果见图3。
表1手性芳杂胺类衍生物的编号及结构
SEQUENCE LISTING
<110> 中山大学
<120> 一种抗人乙肝病毒药物的筛选方法
<130>
<160> 13
<170> PatentIn version 3.3
<210> 1
<211> 192
<212> PRT
<213> HBcAg蛋白序列
<400> 1
Met Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Asp Ile Asp Pro Tyr Lys
1 5 10 15
Glu Phe Gly Ala Thr Val Glu Leu Leu Ser Phe Leu Pro Ser Asp Phe
20 25 30
Phe Pro Ser Val Arg Asp Leu Leu Asp Thr Ala Ser Ala Leu Tyr Arg
35 40 45
Glu Ala Leu Glu Ser Pro Glu His Cys Ser Pro His His Thr Ala Leu
50 55 60
Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu Met Thr Leu Ala Thr Trp
65 70 75 80
Val Gly Val Asn Leu Glu Asp Pro Ala Ser Arg Asp Leu Val Val Ser
85 90 95
Tyr Val Asn Thr Asn Met Gly Leu Lys Phe Arg Gln Leu Leu Trp Phe
100 105 110
His Ile Ser Cys Leu Thr Phe Gly Arg Glu Thr Val Ile Glu Tyr Leu
115 120 125
Val Ser Phe Gly Val Trp Ile Arg Thr Pro Pro Ala Tyr Arg Pro Ser
130 135 140
Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu Thr Cys Val Val Arg Arg
145 150 155 160
Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro Ser Pro Arg Arg Arg Arg
165 170 175
Ser Gln Ser Pro Arg Arg Arg Arg Ser Gln Ser Arg Glu Ser Gln Cys
180 185 190
<210> 2
<211> 154
<212> PRT
<213> YFP-N1 蛋白序列
<400> 2
Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu
1 5 10 15
Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly
20 25 30
Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile
35 40 45
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr
50 55 60
Phe Gly Tyr Gly Leu Gln Cys Phe Ala Arg Tyr Pro Asp His Met Lys
65 70 75 80
Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
85 90 95
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu
100 105 110
Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
115 120 125
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr
130 135 140
Asn Tyr Asn Ser His Asn Val Tyr Ile Met
145 150
<210> 3
<211> 86
<212> PRT
<213> YFP-C1 蛋白序列
<400> 3
Ala Asp Lys Gln Lys Asn Gly Ile Cys Lys Val Asn Phe Lys Ile Arg
1 5 10 15
His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln
20 25 30
Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr
35 40 45
Leu Ser Tyr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp
50 55 60
His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly
65 70 75 80
Met Asp Glu Leu Tyr Lys
85
<210> 4
<211> 172
<212> PRT
<213> YFP-N2 蛋白序列
<400> 4
Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu
1 5 10 15
Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly
20 25 30
Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile
35 40 45
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr
50 55 60
Phe Gly Tyr Gly Leu Gln Cys Phe Ala Arg Tyr Pro Asp His Met Lys
65 70 75 80
Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
85 90 95
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu
100 105 110
Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
115 120 125
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr
130 135 140
Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn
145 150 155 160
Gly Ile Cys Lys Val Asn Phe Lys Ile Arg His Asn
165 170
<210> 5
<211> 68
<212> PRT
<213> YFP-C2 蛋白序列
<400> 5
Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr
1 5 10 15
Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser
20 25 30
Tyr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met
35 40 45
Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp
50 55 60
Glu Leu Tyr Lys
65
<210> 6
<211> 154
<212> PRT
<213> CFP-N1 蛋白序列
<400> 6
Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu
1 5 10 15
Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly
20 25 30
Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile
35 40 45
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr
50 55 60
Leu Thr Trp Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys
65 70 75 80
Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
85 90 95
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu
100 105 110
Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
115 120 125
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr
130 135 140
Asn Tyr Ile Ser His Asn Val Tyr Ile Thr
145 150
<210> 7
<211> 85
<212> PRT
<213> CFP-C1 蛋白序列
<400> 7
Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile Arg His
1 5 10 15
Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn
20 25 30
Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu
35 40 45
Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His
50 55 60
Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met
65 70 75 80
Asp Glu Leu Tyr Lys
85
<210> 8
<211> 172
<212> PRT
<213> CFP-N2 蛋白序列
<400> 8
Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu
1 5 10 15
Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly
20 25 30
Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile
35 40 45
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr
50 55 60
Leu Thr Trp Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys
65 70 75 80
Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
85 90 95
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu
100 105 110
Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
115 120 125
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr
130 135 140
Asn Tyr Ile Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn
145 150 155 160
Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn Ile
165 170
<210> 9
<211> 67
<212> PRT
<213> CFP-C2 蛋白序列
<400> 9
Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro
1 5 10 15
Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr
20 25 30
Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val
35 40 45
Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu
50 55 60
Leu Tyr Lys
65
<210> 10
<211> 154
<212> PRT
<213> VFP-N1 蛋白序列
<400> 10
Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu
1 5 10 15
Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly
20 25 30
Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Leu Ile
35 40 45
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr
50 55 60
Leu Gly Tyr Gly Leu Gln Cys Phe Ala Arg Tyr Pro Asp His Met Lys
65 70 75 80
Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
85 90 95
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu
100 105 110
Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
115 120 125
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr
130 135 140
Asn Tyr Asn Ser His Asn Val Tyr Ile Thr
145 150
<210> 11
<211> 85
<212> PRT
<213> VFP-C1 蛋白序列
<400> 11
Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile Arg His
1 5 10 15
Asn Ile Glu Asp Gly Gly Val Gln Leu Ala Asp His Tyr Gln Gln Asn
20 25 30
Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu
35 40 45
Ser Tyr Gln Ser Lys Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His
50 55 60
Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met
65 70 75 80
Asp Glu Leu Tyr Lys
85
<210> 12
<211> 172
<212> PRT
<213> VFP-N2 蛋白序列
<400> 12
Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu
1 5 10 15
Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly
20 25 30
Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Leu Ile
35 40 45
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr
50 55 60
Leu Gly Tyr Gly Leu Gln Cys Phe Ala Arg Tyr Pro Asp His Met Lys
65 70 75 80
Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
85 90 95
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu
100 105 110
Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
115 120 125
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr
130 135 140
Asn Tyr Asn Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn
145 150 155 160
Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn Ile
165 170
<210> 13
<211> 67
<212> PRT
<213> VFP-C2 蛋白序列
<400> 13
Glu Asp Gly Gly Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro
1 5 10 15
Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Tyr
20 25 30
Gln Ser Lys Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val
35 40 45
Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu
50 55 60
Leu Tyr Lys
65
Claims (5)
1.一种抗人乙肝病毒药物的筛选方法，其特征在于，包括如下步骤：将SEQ ID NO：1所示的HBc分别与荧光蛋白的肽段a和肽段b相连，构建得融合蛋白A和B，所述肽段a和肽段b连接共同组成荧光蛋白的全序列；将融合蛋白A和B通过克隆技术共转化同一受体细胞，用待检测药物处理上述转化的受体细胞，同时以未用药物处理的上述转化的受体细胞作为阴性对照；观察细胞中荧光蛋白的信号，如果无荧光蛋白信号或荧光信号低于阴性对照，说明待检测药物可以抗人乙肝病毒，如果荧光信号与阴性对照信号强度相当，说明待检测药物不可以抗人乙肝病毒。
2.根据权利要求1所述的筛选方法，其特征在在于，所述荧光蛋白为黄色荧光蛋白、蓝绿色荧光蛋白或Venus荧光蛋白。
3.根据权利要求1所述的筛选方法，其特征在在于，所述肽段a的氨基酸序列如SEQ IDNO：2所示，肽段b的氨基酸序列如SEQ ID NO：3所示；或，肽段a的氨基酸序列如SEQ ID NO：4所示，肽段b的氨基酸序列如SEQ ID NO：5所示；或，肽段a的氨基酸序列如SEQ ID NO：6所示，肽段b的氨基酸序列如SEQ ID NO：7所示；或，肽段a的氨基酸序列如SEQ ID NO：8所示，肽段b的氨基酸序列如SEQ ID NO：9所示；或，肽段a的氨基酸序列如SEQ ID NO：10所示，肽段b的氨基酸序列如SEQ ID NO：11所示；或，肽段a的氨基酸序列如SEQ ID NO：12所示，肽段b的氨基酸序列如SEQ ID NO：13所示。
4.根据权利要求1所述的筛选方法，其特征在在于，SEQ ID NO：1所示的HBc分别与荧光蛋白的肽段a和肽段b相连时，使用linker序列连接，所述Linker序列为(GGSG)n；(GGSGG)n；(GSGSG)n；(GSGGG)n；(GGGGS)n；(GGGSG)n；(GSSSG)n；RPACKIPNDLKQKVMNH；RSIAT。
5.根据权利要求1所述的筛选方法，其特征在在于，所述受体细胞为HEK293T细胞、HepG2或Huh7。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611138490.7A CN108611395A (zh) | 2016-12-12 | 2016-12-12 | 一种抗人乙肝病毒药物的筛选方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611138490.7A CN108611395A (zh) | 2016-12-12 | 2016-12-12 | 一种抗人乙肝病毒药物的筛选方法 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108611395A true CN108611395A (zh) | 2018-10-02 |
Family
ID=63643643
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611138490.7A Pending CN108611395A (zh) | 2016-12-12 | 2016-12-12 | 一种抗人乙肝病毒药物的筛选方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108611395A (zh) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109666698A (zh) * | 2018-12-29 | 2019-04-23 | 杭州科兴生物科技有限公司 | 一种基于Tim-3/Galectin-9阻断功能及其生物效应的药物快速筛选方法 |
| CN109666699A (zh) * | 2018-12-29 | 2019-04-23 | 杭州科兴生物科技有限公司 | 一种基于lag-3/mhc ii阻断功能及其生物效应的药物快速筛选方法 |
| CN114751990A (zh) * | 2022-04-01 | 2022-07-15 | 中国科学技术大学 | 酰胺高丝氨酸内酯受体-增强绿色荧光融合蛋白及其制备方法与应用 |
| CN114875052A (zh) * | 2022-06-27 | 2022-08-09 | 齐鲁工业大学 | 一种能够检测透明质酸的融合蛋白及检测方法 |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1015584A1 (en) * | 1997-09-19 | 2000-07-05 | Monsanto Company | Dna vaccination against cholesterol ester transfer protein in the treatment of atherosclerosis |
| EP1165797A2 (de) * | 1999-02-05 | 2002-01-02 | Eberhard Hildt | Partikel zur gentherapie |
| CN104693309A (zh) * | 2015-02-12 | 2015-06-10 | 重庆医科大学 | 重组人乙型肝炎病毒核心蛋白融合蛋白 |
| CN104710534A (zh) * | 2015-02-12 | 2015-06-17 | 重庆医科大学 | 重组HBc融合蛋白 |
| WO2015193484A1 (en) * | 2014-06-20 | 2015-12-23 | Drexel University | Tagged hepadnavirus e antigen and its use in screening antiviral substances |
| CN105367662A (zh) * | 2014-08-29 | 2016-03-02 | 汤正好 | 一种hbv相关的融合蛋白、其制备方法及其应用 |
| CN105380954A (zh) * | 2015-11-03 | 2016-03-09 | 中山大学 | 单宁酸作为hiv-1潜伏感染激活剂的应用 |
| CN106188310A (zh) * | 2016-07-18 | 2016-12-07 | 重庆医科大学 | 分裂的海肾荧光素酶和hbc的融合蛋白及其应用 |
-
2016
- 2016-12-12 CN CN201611138490.7A patent/CN108611395A/zh active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1015584A1 (en) * | 1997-09-19 | 2000-07-05 | Monsanto Company | Dna vaccination against cholesterol ester transfer protein in the treatment of atherosclerosis |
| EP1165797A2 (de) * | 1999-02-05 | 2002-01-02 | Eberhard Hildt | Partikel zur gentherapie |
| WO2015193484A1 (en) * | 2014-06-20 | 2015-12-23 | Drexel University | Tagged hepadnavirus e antigen and its use in screening antiviral substances |
| CN106716128A (zh) * | 2014-06-20 | 2017-05-24 | 豪夫迈·罗氏有限公司 | 加标签的嗜肝DNA病毒e抗原及其在筛选抗病毒物质中的用途 |
| CN105367662A (zh) * | 2014-08-29 | 2016-03-02 | 汤正好 | 一种hbv相关的融合蛋白、其制备方法及其应用 |
| CN104693309A (zh) * | 2015-02-12 | 2015-06-10 | 重庆医科大学 | 重组人乙型肝炎病毒核心蛋白融合蛋白 |
| CN104710534A (zh) * | 2015-02-12 | 2015-06-17 | 重庆医科大学 | 重组HBc融合蛋白 |
| CN105380954A (zh) * | 2015-11-03 | 2016-03-09 | 中山大学 | 单宁酸作为hiv-1潜伏感染激活剂的应用 |
| CN106188310A (zh) * | 2016-07-18 | 2016-12-07 | 重庆医科大学 | 分裂的海肾荧光素酶和hbc的融合蛋白及其应用 |
Non-Patent Citations (3)
| Title |
|---|
| IZZAT FAHIMUDDIN BIN MOHAMED SUFFIAN 等: "Mutation of arginine residues to avoid non-specific cellular uptakes for hepatitis B virus core particles", 《J NANOBIOTECHNOLOGY》 * |
| 刘蕊 等: "M2e-HBC融合蛋白的真核表达", 《陕西医学杂志》 * |
| 骆清铭等: "《生物分子光子学研究前沿》", 31 October 2014, 上海交通大学出版社 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109666698A (zh) * | 2018-12-29 | 2019-04-23 | 杭州科兴生物科技有限公司 | 一种基于Tim-3/Galectin-9阻断功能及其生物效应的药物快速筛选方法 |
| CN109666699A (zh) * | 2018-12-29 | 2019-04-23 | 杭州科兴生物科技有限公司 | 一种基于lag-3/mhc ii阻断功能及其生物效应的药物快速筛选方法 |
| CN109666699B (zh) * | 2018-12-29 | 2023-02-28 | 杭州科兴生物科技有限公司 | 一种基于lag-3/mhc ii阻断功能及其生物效应的药物快速筛选方法 |
| CN109666698B (zh) * | 2018-12-29 | 2023-03-07 | 杭州科兴生物科技有限公司 | 一种基于Tim-3/Galectin-9阻断功能及其生物效应的药物快速筛选方法 |
| CN114751990A (zh) * | 2022-04-01 | 2022-07-15 | 中国科学技术大学 | 酰胺高丝氨酸内酯受体-增强绿色荧光融合蛋白及其制备方法与应用 |
| CN114751990B (zh) * | 2022-04-01 | 2023-10-20 | 中国科学技术大学 | 酰胺高丝氨酸内酯受体-增强绿色荧光融合蛋白及其制备方法与应用 |
| CN114875052A (zh) * | 2022-06-27 | 2022-08-09 | 齐鲁工业大学 | 一种能够检测透明质酸的融合蛋白及检测方法 |
| CN114875052B (zh) * | 2022-06-27 | 2023-11-14 | 齐鲁工业大学 | 一种能够检测透明质酸的融合蛋白及检测方法 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Volz et al. | Modified vaccinia virus Ankara: history, value in basic research, and current perspectives for vaccine development | |
| CN108611395A (zh) | 一种抗人乙肝病毒药物的筛选方法 | |
| Testut et al. | A new hepadnavirus endemic in arctic ground squirrels in Alaska | |
| Freitas et al. | Envelope proteins derived from naturally integrated hepatitis B virus DNA support assembly and release of infectious hepatitis delta virus particles | |
| Bruss | Hepatitis B virus morphogenesis | |
| Prange | Host factors involved in hepatitis B virus maturation, assembly, and egress | |
| Jaoudé et al. | Role of the antigenic loop of the hepatitis B virus envelope proteins in infectivity of hepatitis delta virus | |
| Blum et al. | Naturally occurring missense mutation in the polymerase gene terminating hepatitis B virus replication | |
| Ito et al. | Impairment of hepatitis B virus virion secretion by single-amino-acid substitutions in the small envelope protein and rescue by a novel glycosylation site | |
| Kuroki et al. | A cell surface protein that binds avian hepatitis B virus particles | |
| FI107620B (fi) | Hepatiitti-C-virukseen hybridisoituvia polynukleotideja, niitä sisältäviä välinepakkauksia ja niiden käyttö | |
| Le Pogam et al. | Exposure of RNA templates and encapsidation of spliced viral RNA are influenced by the arginine-rich domain of human hepatitis B virus core antigen (HBcAg 165-173) | |
| Rodriguez-Frias et al. | Quasispecies structure, cornerstone of hepatitis B virus infection: mass sequencing approach | |
| Walewski et al. | Mutation Master: profiles of substitutions in hepatitis C virus RNA of the core, alternate reading frame, and NS2 coding regions | |
| Chouteau et al. | A short N-proximal region in the large envelope protein harbors a determinant that contributes to the species specificity of human hepatitis B virus | |
| Le Pogam et al. | Influence of a putative intermolecular interaction between core and the pre-S1 domain of the large envelope protein on hepatitis B virus secretion | |
| Koenig et al. | Selective infection of human CD4+ cells by simian immunodeficiency virus: productive infection associated with envelope glycoprotein-induced fusion. | |
| Tan et al. | Phage display creates innovative applications to combat hepatitis B virus | |
| Tyagi et al. | Self-association and mapping of the interaction domain of hepatitis E virus ORF3 protein | |
| JPH06172393A (ja) | 宿主炭水化物含量が減少したＨＢｓＡｇエスケープ変異体ワクチン | |
| JPH07503615A (ja) | 肝炎療剤 | |
| CN102112601B (zh) | 新的融合蛋白及其用于制备针对丙型肝炎的疫苗的用途 | |
| Vyas et al. | Hepatitis B virus: Biology, pathogenesis, epidemiology, clinical description, and diagnosis | |
| CN105017410B (zh) | 一种b区部分缺失型重组人凝血因子ⅷ | |
| Pai et al. | Emergence of a novel mutation in the FLLA region of hepatitis B virus during lamivudine therapy |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181002 |
|
| RJ01 | Rejection of invention patent application after publication |