Daclatasvir (BMS-790052)

Catalog No.S1482 Synonyms: EBP883

For research use only.

Daclatasvir (BMS-790052, EBP883) is a highly selective inhibitor of HCV NS5A with EC50 of 9-50 pM, for a broad range of HCV replicon genotypes and the JFH-1 genotype 2a infectious virus in cell culture. Phase 3.

Daclatasvir (BMS-790052) Chemical Structure

CAS No. 1009119-64-5

Selleck's Daclatasvir (BMS-790052) has been cited by 48 publications

Purity & Quality Control

Choose Selective HCV Protease Inhibitors

Other HCV Protease Products

Biological Activity

Description Daclatasvir (BMS-790052, EBP883) is a highly selective inhibitor of HCV NS5A with EC50 of 9-50 pM, for a broad range of HCV replicon genotypes and the JFH-1 genotype 2a infectious virus in cell culture. Phase 3.
Features First-in-class, highly selective inhibitor of hepatitis C virus (HCV) NS5A with picomolar EC50 values.
Targets
HCV NS5A [1]
9 pM-50 pM(EC50)
In vitro

BMS-790052 is one of the most potent inhibitors of HCV replication reported so far. The mean EC50 valuses of BMS-790052 are 50 and 9 pM for HCV genotype 1a and 1b replicons, respectively. BMS-790052 displays a therapeutic index (CC50/EC50) of at least 105 and is inactive towards a panel of 10 RNA and DNA viruses, with EC50 higher than 10 μM. This confirms BMS-790052's specificity for HCV. [1] In Huh7 cells harboring the HCV genotype 1b replicons, BMS-790052 blocks both transient and stable HCV genome replication, with EC50 values raging from 1-15 pM. BMS-790052 (100 pM or 1 nM) has been shown to alter the subcellular localization and biochemical fractionation of NS5A. [2] BMS-790052 inhibits hybrid replicons containing HCV genotype-4 NS5A genes with EC50 of 7-13 pM. Residue 30 of NS5A is an important site for BMS-790052-mediated resistance in the hybrid replicons. [3]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
human CEM cells M{jRdmN6fG:2b4jpZ4l1gSCjc4PhfS=> NXzV[2hHOyCmYYnz M1\JXGN6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJGNGVSClZXzsd{Bi\nSncjCzJIRigXNuIFPDOVA:QS54IN88US=> NVnV[25RRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkWxOVQ4OTRpPkK1NVU1PzF2PD;hQi=>
Huh7 NHf1dmxCdnSrdnnyZYwh[XO|YYm= M1q4OVMh\GG7cx?= NF3BSpBCdnSrdnnyZYwh[WO2aY\peJkh[WejaX7zeEBJS1ZiZ3Xuc5R6eGViMXKgbY5n\WO2ZXSgbY4hcHWvYX6gTJVpPyClZXzsd{Bi\nSncjCzJIRigXNiYomgZ4VtdC2kYYPl[EBz\XCuaXPvckBie3OjeTygSWM2OD1yLkCwNFAxO87:TR?= M2i0[FxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ3MUS4NVAxLz5{NUG0PFExODxxYU6=
Huh-luc/neo-ET replicon MV7BcpRqfmm{YXygZZN{[Xl? MXe0PEBpenN? NHjHNFNCdnSrdnnyZYwh[WO2aY\peJkh[WejaX7zeEBJ\XCjdHn0bZMhSyC4aYL1d{Bo\W6xdInw[UAy[iCqYYLic5JqdmdiTmO1RUBNOjiYIH31eIFvfCCpZX7lJIlvKEi3aD3seYMwdmWxLVXUJJJmeGyrY3;uJINmdGy|IHHmeIVzKDR6IHjyd{BjgSC2cnHud4lmdnRicnXwcIlkd25ibYX0ZY51NWKjc3XkJIx2[2moZYLhd4Uh[XO|YYmsJGVEPTB;MD6wNFAxODUQvF2= NYrXSpF1RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS1Olg{OTNpPkK0OVY5OzF|PD;hQi=>
HuH7 M{K0WGFvfGm4aYLhcEBie3OjeR?= M3;CTWFvfGm4aYLhcEBi[3Srdnn0fUBi\2GrboP0JGhEXiCpZX7veJlx\SBzYjDpcoZm[3SnZDDpckBpfW2jbjDIeWg4KGOnbHzzJIF{e2W|c3XkJIF{KHKnZIXjeIlwdiCrbjD2bZJidCCUTlGgdoVxdGmlYYTpc44tKEWFNUC9NE4xODByMEVOwG0> MXG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPjB7OUWzNkc,OjZyOUm1N|I9N2F-
HuH7 NEizcpZCdnSrdnnyZYwh[XO|YYm= NI[yNpNCdnSrdnnyZYwh[WO2aY\peJkh[WejaX7zeEBJS1ZzYjDpcoZm[3SnZDDpckBpfW2jbjDIeWg4KGOnbHzzJIF{e2W|c3XkJIF{KGmwaHnibZRqd25ib3[geolz[WxicnXwcIlk[XSrb36gZpkhWlRvUFPSJIFv[Wy7c3nzMEBGSzVyPUCuNFAxODB4Nt88US=> NIPISIM9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkWwO|k3OSd-MkK1NFc6PjF:L3G+
HuH-Lcu-Neo NVHpXVJ[TnWwY4Tpc44h[XO|YYm= NESybpg1QCCqcoO= NGrZeotKdmirYnn0bY9vKG:oIF7TOWEhcW5iSFPWJIdmdm:2eYDlJFFjKGmwZnXjeIVlKGmwIFj1TE1N[3VvTnXvJINmdGy|IHHzd4V{e2WmIHHzJJJm\HWldHnvckBqdiC4aYLhcEBz\XCuaXPheIlwdiCrbnP1ZoF1\WRiZn;yJFQ5KGi{czDifUBtfWOrZnXyZZNmKHKncH;yeIVzKGenbnWgZZN{[XluIFXDOVA:OC5yMECwNFgy|ryP MYK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8{OjJyMke4Nkc,OzJ{MEK3PFI9N2F-
Huh-5.2 NWHzNlZ3SW62aY\pdoFtKGG|c3H5 NHSxPWw1KGSjeYO= MX\BcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDI[ZBifGm2aYOgR{B3cXK3czDn[Y5wfHmyZTCxZkBqdm[nY4Tl[EBqdiCqdX3hckBJfWhvNT6yJINmdGy|IHHzd4V{e2WmIHHzJIRm[3KnYYPlJIlvKEiFVjDy[ZBtcWOxbjDSUmEhemWybHnjZZRqd25iYX\0[ZIhPCCmYYnzJIJ6KGy3Y3nm[ZJie2ViYYPzZZktKEWFNUC9NE4xODByMEpOwG0> NWDSN5IzRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS5NFA5OTFpPkK0PVAxQDFzPD;hQi=>
HuH7 M1[2bmFvfGm4aYLhcEBie3OjeR?= M2qxbVMh\GG7cx?= NGnMVoRCdnSrdnnyZYwh[WO2aY\peJkh[WejaX7zeEBJS1ZiZ3Xuc5R6eGViMXKgR49vOSCrbn\lZ5Rm\CCrbjDoeY1idiCKdVi3JINmdGy|IHHzd4V{e2WmIHHzJIlvcGmkaYTpc44hd2ZidnnyZYwhWk6DIILldIxq[2G2aX;uJIFnfGW{IEOg[IF6eyCkeTDy[Y5qdGyjIHz1Z4ln\XKjc3WgdoVxd3K2ZYKg[4Vv\SCjc4PhfUwhTUN3ME2wMlAxODByOd88US=> MVO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDN{MEmzN{c,OjR|MkC5N|M9N2F-
HuH7 NVj6S4JSSW62aY\pdoFtKGG|c3H5 NF;VcWo4OiCqcoO= NVTxVGxnSW62aY\pdoFtKGGldHn2bZR6KGGpYXnud5QhUEOYMXKgbY5n\WO2ZXSgbY4hcHWvYX6gTJVJPyClZXzsd{Bie3Onc4Pl[EBieyCrbnjpZol1cW:wIH;mJJZqemGuIILldIxq[2G2aX;uJIFnfGW{IEeyJIhzeyCkeTDGVmVVKGG|c3H5MEBGSzVyPUCuNFAxODB7zszN MUC8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDV{MUK5PUc,OjR3MkGyPVk9N2F-
HuH7 MWLBcpRqfmm{YXygZZN{[Xl? M3z3SGFvfGm4aYLhcEBi[3Srdnn0fUBi\2GrboP0JGhEXiCpZX7veJlx\SBzYjDDc44yKGmwZnXjeIVlKGmwIHj1cYFvKEi3SEegZ4VtdHNiYYPz[ZN{\WRiYYOgbY5pcWKrdHnvckBw\iC4aYLhcEBz\XCuaXPheIlwdixiRVO1NF0xNjByMECwPe69VQ>? NHr5NFI9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NkC3O|Q6Oyd-Mk[wO|c1QTN:L3G+
Huh7.5/J6/JFH1/EMCVIRES/hRlucNeo MX3GeY5kfGmxbjDhd5NigQ>? MWC3NkBpenN? NEDMfopKdmirYnn0bY9vKG:oIF7TOWEhcW5iSFPWJIdmdm:2eYDlJFJiKGmwZnXjeIVlKGmwIHj1cYFvKEi3aEeuOU9LPi:MRlixM2VOS1[LUlXTM4hTdHWlTnXvJINmdGy|IHHzd4V{e2WmIHHzJIlvcGmkaYTpc44hd2ZicnXwcIlkd25ibHX2[Yx{KGmwY4XiZZRm\CCob4KgO|IhcHK|IHL5JIx2[2moZYLhd4UhemWyb4L0[ZIh\2WwZTDhd5NigSxiSVO1NF0xNjByMECwPe69VQ>? MnK4QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOzF5MUC0O|koRjNzN{GwOFc6RC:jPh?=
HuH7 replicon MkXkSpVv[3Srb36gZZN{[Xl? M2DJZ|Ih\GG7cx?= NVq2eVhDUW6qaXLpeIlwdiCxZjDOV|VCKGmwIFjDWkBo\W6xdInw[UAy[iCrbn\lZ5Rm\CCrbjDoeY1idiCKdVi3JJJmeGyrY3;uJINmdGy|IHHzd4V{e2WmIHHzJJJm\HWldHnvckBqdiC|dXLn[Y5wdWmlII\pdoFtKFKQQTDy[ZBtcWOjdHnvckB1emWjdHXkJIZweiB{IHThfZMh\m:ubH;3[YQh[nliY3;tdI92dmRid3HzbI92fCCjbnSgd5Vje2WzdXXueEBkd22yb4Xu[EBld3OrbnegcYVie3W{ZXSgZYZ1\XJiMTDkZZkh[nliU1XBVEBz\XCxcoTldkBo\W6nLDDFR|UxRTBwMECwNFA6|ryP MU[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8{ODd5Mk[wO{c,OzB5N{K2NFc9N2F-
HuH7 M2TvNGFvfGm4aYLhcEBie3OjeR?= MXfBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3YhOWJiaX7m[YN1\WRiaX6gbJVu[W5iSIXIO{Bk\WyuczDifUBqdiC4aYTyc{Bz\XCuaXPvckBie3OjeTygSWM2OD1yLkCwNFAy|ryP MXe8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzR4NkKzN{c,OjN2Nk[yN|M9N2F-
Huh-luc/neo-ET replicon NX7oSGVDSW62aY\pdoFtKGG|c3H5 M{jxOlQ5KGi{cx?= Mme3RY51cX[rcnHsJIFkfGm4aYT5JIFo[Wmwc4SgTIVx[XSrdHnzJGMhfmm{dYOg[4Vvd3S7cHWgNYIhcW5iSIXoMYx2[y:wZX:tSXQhemWybHnjc44h[2WubIOgZYZ1\XJiNEigbJJ{KGK7IILldIxq[2:wLXLhd4VlKGy3Y3nm[ZJie2ViYYPzZZktKEWFNUC9NE4xODByMd88US=> NX[xdVBnRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS1Olg{OTNpPkK0OVY5OzF|PD;hQi=>
HuH7 M1HKOmZ2dmO2aX;uJIF{e2G7 MlrhTY5pcWKrdHnvckBw\iCQU{XBJIlvKEiFVjDn[Y5wfHmyZT2xZkBqdm[nY4Tl[EBqdiCqdX3hckBJfUh5IHPlcIx{KGK7IHz1Z4ln\XKjc3WgdoVxd3K2ZYKg[4Vv\SCjc4PhfUwhTUN3ME2wMlAxODB{M988US=> MXW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPTR3M{ixNEc,OjV2NUO4NVA9N2F-
HuH7.5/Con1/SG-Neo(I)-hRluc2aUb MVHGeY5kfGmxbjDhd5NigQ>? M1vu[lczKGi{cx?= NEf6R5hKdmirYnn0bY9vKG:oIF7TOWEhcW5iSFPWJIdmdm:2eYDlJFFjKGmwZnXjeIVlKGmwIHj1cYFvKEi3SEeuOU9Ed25zL2PHMW5mdyiLKT3oVox2[zKjVXKgZ4VtdHNiYYPz[ZN{\WRiYYOgbY5pcWKrdHnvckBw\iC{ZYDsbYNwdiCuZY\lcJMhcW6ldXLheIVlKG[xcjC3NkBpenNiYomgcJVkcW[ncnHz[UBz\XCxcoTldkBo\W6nIHHzd4F6NCCLQ{WwQVAvODByMEKz{txO NELvOIc9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9|MUexNFQ4QSd-M{G3NVA1Pzl:L3G+
HuH7 NFziPXlCdnSrdnnyZYwh[XO|YYm= MVfBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3Yh\2Wwb4T5dIUhOWJiSl\IMVEhcW6oZXP0[YQhcW5iaIXtZY4hUHWKNzDj[YxteyCjc4Pld5Nm\CCjczDpcohq[mm2aX;uJI9nKH[rcnHsJJJmeGyrY3H0bY9vNCCHQ{WwQVAvODByMEK4{txO M2rDNFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ4MEe3OFk{Lz5{NkC3O|Q6OzxxYU6=
HuH7 NH3ifY5CdnSrdnnyZYwh[XO|YYm= M3X6cWFvfGm4aYLhcEBi[3Srdnn0fUBi\2GrboP0JGhEXiBzYjDpcoZm[3SnZDDpckBpfW2jbjDIeWg4KGOnbHzzJIJ6KGmwII\peJJwKHKncHzpZ49vKGG|c3H5MEBGSzlyPUCuNFAxODQQvF2= NGSxSW09[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{S2OlI{Oyd-MkO0OlYzOzN:L3G+
HuH-Lcu-Neo MXHGeY5kfGmxbjDhd5NigQ>? MUG0PEBpenN? M37WemlvcGmkaYTpc44hd2ZiTmO1RUBqdiCKQ2[g[4Vvd3S7cHWgNYEhcW6oZXP0[YQhcW5iSIXIMWxkfS2QZX:gZ4VtdHNiYYPz[ZN{\WRiYYOgdoVlfWO2aX;uJIlvKH[rcnHsJJJmeGyrY3H0bY9vKGmwY4XiZZRm\CCob4KgOFghcHK|IHL5JIx2[2moZYLhd4UhemWyb4L0[ZIh\2WwZTDhd5NigSxiRVO1NF0xNjByMECzNlTPxE1? MkDZQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOzJ{MEK3PFIoRjN{MkCyO|gzRC:jPh?=
HuH7 M2jpWGFvfGm4aYLhcEBie3OjeR?= MWrBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3Yh\2Wwb4T5dIUhOWJiZYjwdoV{e2mwZzDOV|VCKEx|MW[gcZV1[W62IHnu[oVkfGWmIHnuJIh2dWGwIFj1TFch[2WubIOgZZN{\XO|ZXSgZZMhemWmdXP0bY9vKGmwII\pdoFtKFKQQTDy[ZBtcWOjdHnvckwhTUN3ME2wMlAxODB|Nd88US=> MlS0QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjZyOUm1N|IoRjJ4MEm5OVMzRC:jPh?=
Huh-luc/neo-ET replicon NIm1RoxCdnSrdnnyZYwh[XO|YYm= MV:0PEBpenN? Mnq4RY51cX[rcnHsJIFkfGm4aYT5JIFo[Wmwc4SgTIVx[XSrdHnzJGMhfmm{dYOg[4Vvd3S7cHWgNYEhcW5iSIXoMYx2[y:wZX:tSXQhemWybHnjc44h[2WubIOgZYZ1\XJiNEigbJJ{KGK7IILldIxq[2:wLXLhd4VlKGy3Y3nm[ZJie2ViYYPzZZktKEWFNUC9NE4xODByM{m4{txO NFHlb|E9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEW2PFMyOyd-MkS1Olg{OTN:L3G+
HuH7 M2rUb2FvfGm4aYLhcEBie3OjeR?= MYjBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDD3bYxlKHS7cHWgTGNXKGenbn;0fZBmKDGkIHnu[oVkfGWmIHnuJIh2dWGwIFj1TFch[2WubIOgZZN{\XO|ZXSgZZMhemWmdXP0bY9vKGmwII\pdoFtKFKQQTDy[ZBtcWOjdHnvckwhTUN3ME2wMlAxODB2ON88US=> MWK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPjB7OUWzNkc,OjZyOUm1N|I9N2F-
W11.8 NFrQXW5CdnSrdnnyZYwh[XO|YYm= NGn5S4M1KGSjeYO= NULSNlVVSW62aY\pdoFtKGGldHn2bZR6KGGpYXnud5QhUGWyYYTpeIl{KENidnnyeZMh\2Wwb4T5dIUhOWFiaX7m[YN1\WRiaX6gW|EyNjhiY3XscJMh[XO|ZYPz[YQh[XNiZHXjdoVie2ViaX6gUnM2SSCneIDy[ZN{cW:wIHnuJJJmeGyrY3;uJINmdGxiYX\0[ZIhPCCmYYnzJIJ6KGy3bXnu[ZNk\W6lZTDiZZNm\CCHTFnTRUwhTUN3ME2wMlAxODB3zszN NV3O[3g{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS5NFA5OTFpPkK0PVAxQDFzPD;hQi=>
HuH7 NWXxZoM2SW62aY\pdoFtKGG|c3H5 MnmwN{Bl[Xm| MnTNRY51cX[rcnHsJIFkfGm4aYT5JIFo[Wmwc4SgTGNXKGenbn;0fZBmKDGjIFi3O{Bqdm[nY4Tl[EBqdiCqdX3hckBJfUh5IHPlcIx{KGG|c3Xzd4VlKGG|IHnubIljcXSrb36gc4Yhfmm{YXygVm5CKHKncHzpZ4F1cW:wIHHmeIVzKDNiZHH5d{BjgSC{ZX7pcIxiKGy3Y3nm[ZJie2VicnXwc5J1\XJiZ3Xu[UBie3OjeTygSWM2OD1yLkCwNFA2|ryP NV7jPYVKRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkSzNlA6OzNpPkK0N|IxQTN|PD;hQi=>
HuH7 NVfKbZZoSW62aY\pdoFtKGG|c3H5 M4G4UVczKGi{cx?= NF\hXYRCdnSrdnnyZYwh[WO2aY\peJkh[WejaX7zeEBJS1ZzYTDpcoZm[3SnZDDpckBpfW2jbjDIeWg4KGOnbHzzJIF{e2W|c3XkJIF{KGmwaHnibZRqd25ib3[geolz[WxicnXwcIlk[XSrb36gZYZ1\XJiN{KgbJJ{KGK7IF\SSXQh[XO|YYmsJGVEPTB;MD6wNFAxPc7:TR?= M4TDdFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NUKxNlk6Lz5{NEWyNVI6QTxxYU6=
HuH7 MnzMRY51cX[rcnHsJIF{e2G7 MWjBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3Yh\2Wwb4T5dIUhPWFiaX7m[YN1\WRiaX6gbJVu[W5iSIXIO{Bk\WyuczDifUBtfWOrZnXyZZNmKHKncH;yeIVzKGenbnWgZZN{[XluIFXDOVA:OC5yMECwOVHPxE1? MVm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPTR3M{ixNUc,OjV2NUO4NVE9N2F-
HuH7 M{fCWWFvfGm4aYLhcEBie3OjeR?= MYjBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3Yh\2Wwb4T5dIUhOWJiaX7m[YN1\WRiaX6gbJVu[W5iSIXIO{Bk\WyuczDifUBtfWOrZnXyZZNmKHKncH;yeIVzKGenbnWgZZN{[XluIFXDOVA:OC5yMECwO|PPxE1? MWe8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPTR3M{ixNUc,OjV2NUO4NVE9N2F-
HuH-Lcu-Neo NFPCOmlHfW6ldHnvckBie3OjeR?= NV;kUGNwPDhiaILz NYrUNlJMUW6qaXLpeIlwdiCxZjDOV|VCKGmwIIDheIlmdnRvZHXybZZm\CCKQ2[g[4Vvd3S7cHWgN4EhcW6oZXP0[YQhcW5iSIXIMWxkfS2QZX:gZ4VtdHNiYYPz[ZN{\WRiYYOgdoVlfWO2aX;uJIlvKH[rcnHsJJJmeGyrY3H0bY9vKGmwY4XiZZRm\CCob4KgOFghcHK|IHL5JIx2[2moZYLhd4UhemWyb4L0[ZIh\2WwZTDhd5NigSxiRVO1NF0xNjByMEGxOFXPxE1? NX3udIR7RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxM{KyNFI4QDJpPkOyNlAzPzh{PD;hQi=>
Huh-luc/neo-ET replicon NH2x[IFCdnSrdnnyZYwh[XO|YYm= M{PHZVQ5KGi{cx?= MWXBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDI[ZBifGm2aYOgR{B3cXK3czDn[Y5wfHmyZTCxZkBp[XKkb4LpcochVlN3QTDMN|FXKG23dHHueEBo\W6nIHnuJGh2cC2udXOvcoVwNUWWIILldIxq[2:wIHPlcIx{KGGodHXyJFQ5KGi{czDifUB1emGwc3nlcpQhemWybHnjc44hdXW2YX70MYJie2WmIHz1Z4ln\XKjc3WgZZN{[XluIFXDOVA:OC5yMECxNlU6|ryP M1ToN|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NU[4N|E{Lz5{NEW2PFMyOzxxYU6=
HuH7 MVnGeY5kfGmxbjDhd5NigQ>? NXrHUWZYUW6qaXLpeIlwdiCxZjDOV|VCKGmwIFjDWkBo\W6xdInw[U0y[SCrbn\lZ5Rm\CCrbjDoeY1idiCKdVi3JINmdGy|IHL5JIx2[2moZYLhd4UhemWyb4L0[ZIh\2WwZTDhd5NigSxiRVO1NF0xNjByMEG0{txO M{ftVVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ3NEWzPFExLz5{NUS1N|gyODxxYU6=
HuH7 M17DU2FvfGm4aYLhcEBie3OjeR?= NF3mXm9CdnSrdnnyZYwh[WO2aY\peJkh[WejaX7zeEBJS1ZiZ3Xuc5R6eGViMXGgbY5n\WO2ZXSgbY4hcHWvYX6gTJVJPyClZXzsd{BjgSCudXPp[oVz[XOnIILldI9zfGW{IHflcoUh[XO|YYmsJGVEPTB;MD6wNFAyPM7:TR?= NHrqcYs9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NUS1N|gyOSd-MkW0OVM5OTF:L3G+
HuH7 MofURY51cX[rcnHsJIF{e2G7 MlX3RY51cX[rcnHsJIFkfGm4aYT5JIFo[Wmwc4SgTGNXKGenbn;0fZBmKDGkIHX4dJJme3OrbnegUnM2SSC\OUPIJI12fGGwdDDpcoZm[3SnZDDpckBpfW2jbjDIeWg4KGOnbHzzJIF{e2W|c3XkJIF{KHKnZIXjeIlwdiCrbjD2bZJidCCUTlGgdoVxdGmlYYTpc44tKEWFNUC9NE4xODBzON88US=> MYW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPjB7OUWzNkc,OjZyOUm1N|I9N2F-
Huh-luc/neo-ET replicon MlnLRY51cX[rcnHsJIF{e2G7 NYHpUHo4PDhiaILz M361emFvfGm4aYLhcEBi[3Srdnn0fUBi\2GrboP0JGhmeGG2aYTpd{BEKH[rcoXzJIdmdm:2eYDlJFFjKGijcnLvdolv\yCQU{XBJHk6O0hibYX0ZY51KGenbnWgbY4hUHWqLXz1Z{9v\W9vRWSgdoVxdGmlb36gZ4VtdHNiYX\0[ZIhPDhiaILzJIJ6KHS{YX7zbYVvfCC{ZYDsbYNwdiCvdYThcpQu[mG|ZXSgcJVkcW[ncnHz[UBie3OjeTygSWM2OD1yLkCwNFMyPjMQvF2= MkXRQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR3NkizNVMoRjJ2NU[4N|E{RC:jPh?=
HuH7 M4nieWFvfGm4aYLhcEBie3OjeR?= M1To[lczKGi{cx?= MUDBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3Yy[SCrbn\lZ5Rm\CCrbjDoeY1idiCKdVi3JINmdGy|IHHzd4V{e2WmIHHzJIlvcGmkaYTpc44hd2ZidnnyZYwhemWybHnjZZRqd25iYX\0[ZIhPzJiaILzJIJ6KE[URWSgZZN{[XluIFXDPVA:OC5yMECzPO69VQ>? M1vuN|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NUKxNlk6Lz5{NEWyNVI6QTxxYU6=
HuH7 NUnGVGRKSW62aY\pdoFtKGG|c3H5 MX;BcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3Yh\2Wwb4T5dIUhPGFiaX7m[YN1\WRiaX6gbJVu[W5iSIXIO{Bk\WyuczDifUBtfWOrZnXyZZNmKHKncH;yeIVzKGenbnWgZZN{[XluIFXDOVA:OC5yMEC0Ne69VQ>? M{LkVVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ3NEWzPFEyLz5{NUS1N|gyOTxxYU6=
HuH-Lcu-Neo M2DZU2Z2dmO2aX;uJIF{e2G7 MYi0PEBpenN? MVTJcohq[mm2aX;uJI9nKE6VNVGgbY4hUEOYIHflco91gXCnIEPhJINwdiCrbn\lZ5Rm\CCrbjDIeWguVGO3LV7lc{Bk\WyuczDhd5Nme3OnZDDhd{Bz\WS3Y4Tpc44hcW5idnnyZYwhemWybHnjZZRqd25iaX7jeYJifGWmIH\vdkA1QCCqcoOgZpkhdHWlaX\ldoF{\SC{ZYDvdpRmeiCpZX7lJIF{e2G7LDDFR|UxRTBwMECwOFEyPc7:TR?= NIX6UJU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9|MkKwNlc5Oid-M{KyNFI4QDJ:L3G+
HuH7 NV7XfJh4SW62aY\pdoFtKGG|c3H5 NGDSSZVCdnSrdnnyZYwh[WO2aY\peJkh[WejaX7zeEBJS1ZiZ3Xuc5R6eGViNnGgbY5n\WO2ZXSgbY4hcHWvYX6gTJVJPyClZXzsd{BjgSCudXPp[oVz[XOnIILldI9zfGW{IHflcoUh[XO|YYmsJGVEPTB;MD6wNFA1Pc7:TR?= NIfUb3k9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NUS1N|gyOSd-MkW0OVM5OTF:L3G+
HuH7 MWjBcpRqfmm{YXygZZN{[Xl? M1zXcmFvfGm4aYLhcEBi[3Srdnn0fUBi\2GrboP0JGhEXiCpZX7veJlx\SB|YTDpcoZm[3SnZDDpckBpfW2jbjDIeWg4KGOnbHzzJIJ6KGy3Y3nm[ZJie2VicnXwc5J1\XJiZ3Xu[UBie3OjeTygSWM2OD1yLkCwNFY4|ryP NE\Db2Y9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NUS1N|gyOSd-MkW0OVM5OTF:L3G+
HuH7 MmX5RY51cX[rcnHsJIF{e2G7 M4jkfWFvfGm4aYLhcEBi[3Srdnn0fUBi\2GrboP0JGhEXiCpZX7veJlx\SB{YTDpcoZm[3SnZDDpckBpfW2jbjDIeWg4KGOnbHzzJIJ6KGy3Y3nm[ZJie2VicnXwc5J1\XJiZ3Xu[UBie3OjeTygSWM2OD1yLkCwNFY4|ryP M4ewWFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ3NEWzPFEyLz5{NUS1N|gyOTxxYU6=
HuH-Lcu-Neo M2rF[2Z2dmO2aX;uJIF{e2G7 NIHqWpE1QCCqcoO= M1S4WGlvcGmkaYTpc44hd2ZiTmO1RUBqdiCyYYTp[Y51NWSncnn2[YQhUEOYIHflco91gXCnIELhJIlv\mWldHXkJIlvKEi3SD3MZ5UuVmWxIHPlcIx{KGG|c3Xzd4VlKGG|IILl[JVkfGmxbjDpckB3cXKjbDDy[ZBtcWOjdHnvckBqdmO3YnH0[YQh\m:{IES4JIhzeyCkeTDseYNq\mW{YYPlJJJmeG:{dHXyJIdmdmViYYPzZZktKEWFNUC9NE4xPDl2zszN MWC8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8{OjJyMke4Nkc,OzJ{MEK3PFI9N2F-
HuH-Lcu-Neo NYruXFJHTnWwY4Tpc44h[XO|YYm= NI[xeFM1QCCqcoO= NGfoUGtKdmirYnn0bY9vKG:oIF7TOWEhcW5iSFPWJIdmdm:2eYDlJFJiKGOxbjDpcoZm[3SnZDDpckBJfUhvTHP1MW5mdyClZXzsd{Bie3Onc4Pl[EBieyC{ZXT1Z5Rqd25iaX6geolz[WxicnXwcIlk[XSrb36gbY5kfWKjdHXkJIZweiB2ODDodpMh[nlibIXjbYZmemG|ZTDy[ZBwenSncjDn[Y5mKGG|c3H5MEBGSzVyPUCuNFUzQDYQvF2= MW[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8{OjJyMke4Nkc,OzJ{MEK3PFI9N2F-
CEM MnPPR5l1d3SxeHnjbZR6KGG|c3H5 M{LhR|Mh\GG7cx?= MX7DfZRwfG:6aXPpeJkh[WejaX7zeEBpfW2jbjDDSW0h[2WubIOgZYZ1\XJiMzDkZZl{NCCFQ{WwQVkvPs7:TR?= NYj6ZlRxRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK1NFc6PjFpPkKyOVA4QTZzPD;hQi=>
CEM NHzGSJhEgXSxdH;4bYNqfHliYYPzZZk> Ml\hR5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gR2VOKGOnbHzzMEBESzVyPUmuOu69VQ>? NULQUmVKRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK3NFQ5QDdpPkKyO|A1QDh5PD;hQi=>
Vero NFGwfYxEgXSxdH;4bYNqfHliYYPzZZk> NH3yV2JEgXSxdH;4bYNqfHliYXfhbY5{fCCjZoLpZ4FvKGe{ZXXuJI1wdmuneTDW[ZJwKGOnbHzzMEBESzVyPUmuOu69VQ>? NVTHbFVmRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkO0OlYzOzNpPkKzOFY3OjN|PD;hQi=>
CEM MV7DfZRwfG:6aXPpeJkh[XO|YYm= NWfxUXRWS3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hS0WPIHPlcIx{NCCFQ{WwQVEx|ryP NUjHVXNlRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMk[wPVk2OzJpPkK2NFk6PTN{PD;hQi=>
PBMC NULIVIpES3m2b4TvfIlkcXS7IHHzd4F6 NYGzbpJoS3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hWEKPQzDj[YxteyxiQ1O1NF0yQc7:TR?= M3m0[FxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{N{C0PFg4Lz5{MkewOFg5PzxxYU6=
Vero NWnKeodkS3m2b4TvfIlkcXS7IHHzd4F6 M3q3PFMh\GG7cx?= NHntSpFEgXSxdH;4bYNqfHliYXfhbY5{fCCjZoLpZ4FvKGe{ZXXuJI1wdmuneTDW[ZJwKGOnbHzzJIFnfGW{IEOg[IF6eyxiQ1O1NF0zOc7:TR?= NVPPfWtYRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK1NFc6PjFpPkKyOVA4QTZzPD;hQi=>
Vero M4XPbWN6fG:2b4jpZ4l1gSCjc4PhfS=> NGHLdHlEgXSxdH;4bYNqfHliYXfhbY5{fCCjZoLpZ4FvKGe{ZXXuJI1wdmuneTDW[ZJwKGOnbHzzMEBESzVyPUKx{txO NVvHZ4tJRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK3NFQ5QDdpPkKyO|A1QDh5PD;hQi=>
CEM NEnNOHhEgXSxdH;4bYNqfHliYYPzZZk> NWrrSlJ5S3m2b4TvfIlkcXS7IHHnZYlve3RiaIXtZY4hS0WPIHPlcIx{NCCFQ{WwQVIy|ryP Mn7GQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjN2Nk[yN|MoRjJ|NE[2NlM{RC:jPh?=
Vero M{Tnb2N6fG:2b4jpZ4l1gSCjc4PhfS=> NGTNdHNEgXSxdH;4bYNqfHliYXfhbY5{fCCjZoLpZ4FvKGe{ZXXuJI1wdmuneTDW[ZJwKGOnbHzzMEBESzVyPUKx{txO NITHc3c9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NkC5PVU{Oid-Mk[wPVk2OzJ:L3G+
Huh7.5.1 NIDEdXlCdnSrdnnyZYwh[XO|YYm= NV;KSnZrOTByIIDNJJRwKDFidV2= MVzBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3Yh\2Wwb4T5dIUhOWJiTluvVlJCViCrbn\lZ5Rm\CCrbjDoeY1idiCKdXi3MlUvOSClZXzsd{BmgHC{ZYPzbY5oKE6VNVGgUFMyXiCvdYThcpQh[XO|ZYPz[YQh[XNicnXkeYN1cW:wIHnuJJZqenW|IILldIxq[2G2aX;uJIF1KDFyMDDwUUB1dyBzIIXNJIJ6KGy3Y3nm[ZJie2VicnXwc5J1\XJiZ3Xu[UBie3OjeR?= NVn0SlQ{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMk[xN|Q2PTFpPkK2NVM1PTVzPD;hQi=>
Huh7.5.1 NV3YUFR4SW62aY\pdoFtKGG|c3H5 MXuxNFAheE1idH:gNUB2VQ>? M{D4Z2FvfGm4aYLhcEBi[3Srdnn0fUBi\2GrboP0JGhEXiCpZX7veJlx\SBzYjDOT{9TOkGQIHnu[oVkfGWmIHnuJIh2dWGwIFj1bFcvPS5zIHPlcIx{KGW6cILld5NqdmdiTmO1RUB[QTOKIH31eIFvfCCjc4Pld5Nm\CCjczDy[YR2[3Srb36gbY4hfmm{dYOgdoVxdGmlYYTpc44h[XRiMUCwJJBOKHSxIEGgeW0h[nlibIXjbYZmemG|ZTDy[ZBwenSncjDn[Y5mKGG|c3H5 MkjiQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjZzM{S1OVEoRjJ4MUO0OVUyRC:jPh?=
GS4.3 MU\BcpRqfmm{YXygZZN{[Xl? MmHRNE4yPSC3TR?= MYG2JIRigXN? MnjFRY51cX[rcnHsJIFkfGm4aYT5JIFo[Wmwc4SgTGNXKGmwZnXjeIVlKGmwIHj1cYFvKEeVND6zJINmdGy|IHHzd4V{e2WmIHHzJIlvcGmkaYTpc44hd2ZiTmOzM|RCKGyndnXsd{BifCByLkG1JJVOKHS{ZXH0[YQhf2m2aDDmdoV{cCCvZXTpZUBkd262YXnubY5oKGOxbYDveY5lKGW4ZYL5JFIh\GG7czDt[YF{fXKnZDDh[pRmeiB4IHThfZMh[nliV3XzeIVzdiCkbH;0JI1mfGixZB?= M1rBVlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ7MkOyOVgzLz5{OUKzNlU5OjxxYU6=
GS4.3 M1;DfWFvfGm4aYLhcEBie3OjeR?= MorlNE4yPSC3TR?= NH;mOYY3KGSjeYO= MVXBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3YhcW6oZXP0[YQhcW5iaIXtZY4hT1N2LkOgZ4VtdHNiYYPz[ZN{\WRiYYOgbY5pcWKrdHnvckBw\iC4aYLhcEBkd3KnIIDyc5RmcW5ibHX2[Yx{KGG2IECuNVUhfU1idILlZZRm\CC5aYToJIZz\XOqIH3l[IliKGOxboThbY5qdmdiY3;tdI92dmRiZY\ldpkhOiCmYYnzJI1m[XO3cnXkJIFnfGW{IE[g[IF6eyCkeTDX[ZN1\XKwIHLsc5QhdWW2aH;k M2T2e|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ7MkOyOVgzLz5{OUKzNlU5OjxxYU6=
HuH7 NGr1e4ZCdnSrdnnyZYwh[XO|YYm= M2rDbFMh\GG7cx?= MUTBcpRqfmm{YXygZYN1cX[rdImgZYdicW6|dDDIR3Yh\2Wwb4T5dIUhOWJiaX7m[YN1\WRiaX6gbJVu[W5iSIXIO{Bk\WyuczDheEA,RSBzMDD0bY1meyCjboTpeolz[WxiRVO1NEBi\nSncjCzJIRigXNiYomgcJVkcW[ncnHz[UBz\XCxcoTldkBie3OjeR?= NHK4doQ9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{OESzNFQ{Pyd-Mki0N|A1Ozd:L3G+
HuH7 NFq3R3FCdnSrdnnyZYwh[XO|YYm= M4HkdVMh\GG7cx?= NXPORlQ6SW62aY\pdoFtKGGldHn2bZR6KGGpYXnud5QhUEOYIHflco91gXCnIEHiJIlv\mWldHXkJIlvKGi3bXHuJGh2UDdiY3XscJMh[29vdILlZZRm\CC5aYToJIF{fW6jcILleolzKGGodHXyJFMh\GG7czDifUBtfWOrZnXyZZNmKHKncH;yeIVzKGG|c3H5 NUjXSJU6RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMki0N|A1OzdpPkK4OFMxPDN5PD;hQi=>

Protocol (from reference)

Kinase Assay:[4]
  • FRET assay for HCV NS5A inhibitors:

    The peptide (Ac-Asp-Glu-Asp [EDANS]-Glu-Glu-Abu-[COO] Ala-Ser-Lys [DABCYL]-NH2) contains a fluorescence donor {EDANS, 5-[(2-aminoethyl)amino]naphthalene-1-sulfonic acid} near one end of the peptide and an acceptor {DABCYL, 4-[(4-dimethylamino)phenyl]azo)benzoic acid} near the other end. Intermolecular resonance energy transfer between the donor and the acceptor quenches the fluorescence of the peptide, but as the NS3 protease cleaves the peptide, the products are released from resonance energy transfer quenching. The fluorescence of the donor increases over time as more substrate is cleaved by the NS3 protease. The assay reagent is: 5× luciferase cell culture lysis reagent diluted to 1× with dH2O, NaCl (150 mM), the FRET peptide (20 μM). HCV-Huh-7 cells are placed in a 96-well plate, and allowed to attach overnight (1×104 cells per well). The next day, BMS-790052 is added to the wells and the plate is incubated for 72 hours. The plate is then rinsed with PBS and used for the FRET assay by the addition of 30 μL of the FRET peptide assay reagent (described above) per well. Signals are obtained using the Cytofluor 4000 instrument, which has been set to 340 nm (excitation)/490 nm (emission) automatic mode, for 20 cycles or less, with the plate being read in the kinetic mode. Following FRET, 40 μL of luciferase substrate is added to each well and the luciferase is measured.

Cell Research:[1]
  • Cell lines: HCV replicon cells (Huh7)
  • Concentrations: 0.1 pM - 50 μM, dissolved in DMSO (the final concentration of DMSO is 0.5%)
  • Incubation Time: 72 hours
  • Method: BMS-790052 is added to 96-well plates containing HCV replicon cells seeded approximately 12 hours before in 200 µL media.The cell plates are tested for replication activity and cytotoxicity after 72 hours of incubation. Cytotoxicity is measured with CellTiter-Blue, after which the media and dye are removed, plates are inverted and the remaining liquid is blotted with paper towels. Replication activity of the HCV genotype 1a cell lines is quantified using Renilla luciferase. 1× Renilla luciferase lysis buffer (30 µL) is added to each well and plates are incubated with gentle shaking for 15 min. Renilla luciferase substrate (40 µL) is then added and the signals are detected using a Top Count luminometer set for light emission quantification. One hundred per cent activity is calculated for each cell line for the DMSO-only wells; percentage activity is calculated for each concentration of the inhibitor by dividing the average value for wells containing compound by the average value for wells containing DMSO.

Solubility (25°C)

In vitro

Chemical Information

Molecular Weight 738.88
Formula

C40H50N8O6

CAS No. 1009119-64-5
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles CC(C)C(C(=O)N1CCCC1C2=NC=C(N2)C3=CC=C(C=C3)C4=CC=C(C=C4)C5=CN=C(N5)C6CCCN6C(=O)C(C(C)C)NC(=O)OC)NC(=O)OC

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Clinical Trial Information

NCT Number Recruitment Interventions Conditions Sponsor/Collaborators Start Date Phases
NCT04852614 Recruiting Diagnostic Test: Pharmacokinetic test Hepatitis C Virus Infection Ain Shams University December 1 2020 --
NCT04773756 Completed Drug: Sofosbuvir 400 MG/ Daclatasvir 60mg Covid19 Alexandria University November 1 2020 Phase 4
NCT03208322 Unknown status Other: Non-Interventional Hepatitis C Bristol-Myers Squibb November 30 2018 --
NCT03487848 Terminated Drug: Daclatasvir|Drug: Sofosbuvir Hepatitis C|Chronic Hepatitis Bristol-Myers Squibb June 25 2018 Phase 2
NCT03500562 Terminated -- Chronic Hepatitis C Bristol-Myers Squibb May 8 2018 --

(data from https://clinicaltrials.gov, updated on 2022-01-17)

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
If you have any other enquiries, please leave a message.

* Indicates a Required Field

Please enter your name.
Please enter your email. Please enter a valid email address.
Please write something to us.
Tags: buy Daclatasvir (BMS-790052) | Daclatasvir (BMS-790052) supplier | purchase Daclatasvir (BMS-790052) | Daclatasvir (BMS-790052) cost | Daclatasvir (BMS-790052) manufacturer | order Daclatasvir (BMS-790052) | Daclatasvir (BMS-790052) distributor