IP Library › Granted Patent US 12,460,244
Granted Patent B2
US 12,460,244 · App. 17/269,995 · Granted Nov 4, 2025

Inhibitors of RNA-guided nuclease activity and uses thereof

Inventors: Amit Choudhary (Boston, MA); Kurt Cox (Cambridge, MA)
Assignees: The Broad Institute, Inc.; The Brigham and Women's Hospital, Inc.
C12Q1/44A61K31/397C12N9/22G01N2333/922G01N2500/00
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Quick Facts
Patent No.
US 12,460,244
App. No.
17/269,995
Granted
Nov 4, 2025
Kind
B2
Abstract

Compositions and methods are provided for the inhibition of the function of RNA guided endonucleases, including the identification and use of such inhibitors. Methods of identifying inhibitor compounds of RNA guided nucleases, assays for detecting nuclease activity and compounds for use therein are also provided.

Claims (24)

1 . A method of inhibiting the nuclease activity of an RNA-guided endonuclease, the comprising contacting an RNA-guided endonuclease-guide complex with a compound of Formula I:

wherein:

R 1 is hydrogen;

R 2 is selected from hydrogen,

 and aryl optionally substituted with one or more R 10 ;

R 3 is hydrogen or C 1 -C 3 alkyl;

R 4 is hydrogen or C 1 -C 3 alkyl;

R 5 is C 1 -C 3 alkyl or C 3 -C 8 cycloalkyl each optionally substituted with one or more R 11 ;

each R 10 is independently selected from halogen, C 1 -C 3 alkyl, haloalkyl, and alkoxy;

each R 11 is independently selected from halogen, C 1 -C 3 alkyl, haloalkyl, and alkoxy; and

X is C 1 -C 3 alkylalcohol.

2 . The method of claim 1 , wherein the compound of Formula I:

3 . The method of claim 1 , wherein the compound of Formula I:

4 . The method of claim 1 , wherein the compound of Formula I is:

5 . The method of claim 1 , wherein the compound of Formula I is:

6 . The method of claim 1 , wherein the method is an in vitro method or an in vivo method.

7 . The method of claim 1 , wherein the method is performed in a cell.

8 . The method of claim 7 , wherein the cell is a prokaryotic cell or a eukaryotic cell.

9 . The method of claim 8 , wherein the prokaryotic cell is a bacterium.

10 . The method of claim 8 , wherein the eukaryotic cell is a human cell, a mammalian cell, an insect cell, a plant cell, or a yeast cell.

11 . The method of claim 7 , wherein the cell is in an organism.

12 . The method of claim 11 , wherein the organism is a human, mammal, vertebrate, invertebrate, insect, or plant.

13 . The method of claim 1 , wherein the RNA-guided nuclease is Cas9; optionally wherein the RNA-guided nuclease is SpCas9 or SaCas9.

14 . The method of any one of claim 1 , wherein the RNA-guided nuclease is Cas12a; optionally wherein the RNA-guided nuclease is FnCas12a.

Assignments (3)
CONFIRMATORY LICENSE Recorded Nov 22, 2023
From: BROAD INSTITUTE, INC.
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 065664/0633 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 19, 2021
From: CHOUDHARY, AMIT
To: THE BRIGHAM AND WOMEN'S HOSPITAL, INC.
Reel/Frame 055345/0770 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 19, 2021
From: COX, KURT
To: THE BROAD INSTITUTE, INC.
Reel/Frame 055345/0793 →
Continuity (3)
Provisional Application 62773694 · Nov 30, 2018
Provisional Application 62765352 · Aug 20, 2018
Related Publication 20210355522A1 · Nov 18, 2021
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