Methods for treating hepatitis b virus infections using ns5a, ns5b or ns3 inhibitors
A technology of hepatitis B virus and hepatitis C virus, which is applied in the field of treatment of hepatitis B virus infection, and can solve the problem of not being able to clear HBV infection
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Embodiment 1
[0900] Example 1: Antiviral activity
[0901] Another aspect of the present disclosure pertains to methods of inhibiting viral infection comprising the step of treating a sample or subject suspected of being in need of such inhibition with a composition of the present disclosure.
[0902] In the disclosed context, samples suspected of containing viruses include natural or man-made materials, such as living organisms; tissue or cell cultures; biological samples, such as samples of biological material (blood, serum, urine, cerebrospinal fluid, tears, sputum fluid, saliva, tissue samples, etc.); laboratory samples; food, water or air samples; biological product samples such as cell extracts, especially recombinant cells that synthesize the desired glycoprotein; etc. Typically, the sample is suspected of containing an organism that induces viral infection, usually a pathogenic organism such as a tumor virus. Samples can be contained in any medium, including water and organic solven...
Embodiment 2
[0905] Example 2: Cell-Based Anti-HCV Activity Evaluation
[0906] To evaluate the ability of compounds to inhibit NS5A, NS5B or NS3, the following protocol can be used.
[0908] Antiviral potency (EC) was determined using a Renilla luciferase (RLuc)-based HCV replicon reporter gene assay. 50 ). For genotype 1 and 2a JFH-1 assays, stable HCV1a RLuc replicon cells (carrying a bicistronic genotype 1a H77 replicon encoding the RLuc reporter gene), stable HCV 1b RLuc replicon cells ( Dicistronic genotype 1b Con1 replicon encoding RLuc reporter gene) or stable HCV 2a JFH-1Rluc replicon cells (dicistronic genotype 2a JFH-1 replicon encoding RLuc reporter gene; L31 is present in NS5A) dispensed into 384-well plates for EC 50 Determination. For genotype 2a (M31 is present in NS5A) or genotype 2b assays, the reporter gene encoding RLuc-Neo and the NS5A gene encoding the genotype 2a J6 strain or the genotype 2b MD2b-1 strain NS5A gene (both with M31 ...
Embodiment 3
[0919] Example 3. Reduction of hepatitis B surface antigen
[0920] The inhibitors described herein were tested for their ability to reduce hepatitis B surface antigen (HBsAg) based on the following protocol.
[0921] HBV cell line
[0922] HepG2.2.15 cell (Acs etc., Proc Natl Acad Sci USA, 84, (1987), 4641-4) is a kind of constitutive HBV expression cell line, and it in DMEM+Glutamax-I medium (Invitrogen, Carlsbad, CA, USA), the culture medium was supplemented with 10% fetal bovine serum (Invitrogen) and G418 (Invitrogen), the final concentration was 200mg / L, and kept at 37°C in 5% CO 2 middle.
[0923] HBsAg determination
[0924] HepG2.2.15 cells were seeded in duplicate into white 96-well plates at 1.5 × 10 cells / well. Cells were treated with a three-fold serial dilution series of the compound in DMSO. The final DMSO concentration in all wells was 1%, and DMSO was used as a no drug control.
[0925] The HBsAg Chemiluminescent Immunoassay (CLIA) kit (Autobio Diagnosti...
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