HBV RNAse H purification and enzyme inhibitors

Patent metadata

Field	Value
Publication number	US-10329542-B2
Application number	US-201715670008-A
Country	US
Kind code	B2
Filing date	Aug 7, 2017
Priority date	Nov 27, 2012
Publication date	Jun 25, 2019
Grant date	Jun 25, 2019

Abstract

Provided herein are methods for the obtention of an active HBV RNaseH preparation and its use in screening methods to identify potential inhibitors of the enzyme for possible use as therapeutic agents. Also provided are methods of treatment using agents identified according to the screen.

First claim

What is claimed:

1. A method of inhibiting a hepatitis B virus (HBV) RNAseH comprising contacting said enzyme with a compound having the formula:

wherein:
R 11 is hydrogen, —OR 17 , hydroxy, or halo, wherein R 17 is C 1 -C 8 acyl, C 1 -C 8 alkyl, or a substituted version of either of these groups;
R 12 is hydrogen, hydroxy or halo;
R 13 and R 14 are each independently hydrogen, hydroxy, nitroso, C 1 -C 12 alkyl, C 1 -C 12 alkenyl, C 1 -C 12 alkynyl, C 1 -C 12 aryl, C 1 -C 12 aralkyl, C 1 -C 12 amido, a substituted version of any of these groups, or R 13 is taken together with R 14 as provided below;
R 15 is hydrogen, C 1 -C 12 alkyl, C 1 -C 12 alkenyl, C 1 -C 12 alkynyl, C 1 -C 12 aryl, C 1 -C 12 aralkyl,

or a substituted version of any of these groups; and
R 16 is hydrogen, unsubstituted C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, or hydroxyl;
provided that when R 13 and R 14 are taken together as further defined by FORMULA IV:

R 18 and R 19 are each independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 alkenyl, C 1 -C 6 alkynyl.

2. The method of claim 1 , wherein the compound is of FORMULA III and R 11 is hydroxy.

3. The method of claim 1 , wherein the compound is of FORMULA III and at least one of R 12 , R 13 , R 14 or R 15 is C 1 -C 12 -alkyl.

4. The method of claim 1 , wherein the compound is of FORMULA IV and R 18 is C 1 -C 6 alkenyl.

5. The method of claim 1 , further comprising contacting said enzyme with a second inhibitor of RNAse enzyme activity.

6. The method of claim 5 , wherein said second inhibitor is a nucleoside analog.

7. The method of claim 1 , further comprising contacting said enzyme with said compound a second time.

8. The method of claim 1 , wherein said enzyme is located in a cell.

9. The method of claim 8 , wherein said cell is located in vitro.

10. The method of claim 8 , wherein said cell is located in a living subject.

11. The method of claim 10 , wherein said subject is a mammal infected with HBV.

12. The method of claim 11 , wherein said compound is administered intravenously, intra-arterially, orally, or subcutaneously.

13. The method of claim 11 , wherein said subject is further administered a second inhibitor of RNAse enzyme activity.

14. The method of claim 13 , wherein said second inhibitor is a nucleoside analog.

15. The method of claim 13 , wherein said second inhibitor is administered to said subject before or after said compound.

16. The method of claim 13 , wherein said second inhibitor is administered to said subject at the same time as said compound.

17. The method of claim 10 , wherein said subject has previously received a first-line HBV therapy.

18. The method of claim 17 , wherein said HBV has developed resistance to said first-line HBV therapy.

19. The method of claim 11 , wherein said subject is administered interferon or pegylated interferon.

20. The method of claim 1 , wherein the compound is further defined as:

or a pharmaceutically acceptable salt thereof.

Assignees

• Univ Saint Louis

Inventors

• Tavis John
• Hu Yuan

Classifications

• C12N9/12 Primary

  transferring phosphorus containing groups, e.g. kinases (2.7) · CPC title

• C12Q1/485

  involving kinase · CPC title

• C12N9/22

  Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} · CPC title

• A61K31/519

  ortho- or peri-condensed with heterocyclic rings · CPC title

• C12Q1/706

  for hepatitis · CPC title