Polymerase inhibitors and related compositions and methods

Patent metadata

Field	Value
Publication number	US-11524946-B2
Application number	US-201916712098-A
Country	US
Kind code	B2
Filing date	Dec 12, 2019
Priority date	Dec 12, 2018
Publication date	Dec 13, 2022
Grant date	Dec 13, 2022

Abstract

The present disclosure includes compositions and methods for improved DNA amplification reactions. In particular, the present disclosure provides compositions and methods for hot-start PCR applications using DNA polymerase inhibitors that minimize non-specific DNA amplification by inactivating DNA polymerase at lower temperatures.

First claim

What is claimed is:

1. A compound of formula (I):

or a salt thereof,
wherein:
A is selected from aryl, heteroaryl, and heterocyclyl, each of which may be optionally substituted with 1, 2, or 3 substituents;
R 1 is C 6 -C 20 alkyl;
R 2 is selected from hydrogen and —COOH; and
n is 1 or 2.

2. The compound of claim 1 , or a salt thereof, wherein A is phenyl that is unsubstituted or substituted with 1, 2, or 3 substituents independently selected from C 1 -C 4 alkyl, C 1 -C 4 alkoxy, and halo.

3. The compound of claim 1 , or a salt thereof, wherein A is a 5- or 6-membered monocyclic heteroaryl that is unsubstituted or substituted with 1, 2, or 3 substituents independently selected from C 1 -C 4 alkyl, C 1 -C 4 alkoxy, and halo.

4. The compound of claim 1 , or a salt thereof, wherein A is a bicyclic heterocyclyl group that is unsubstituted.

5. The compound of claim 1 , or a salt thereof, wherein R 1 is C 8 -C 14 alkyl.

6. The compound of claim 1 , wherein the compound is selected from:

or a salt thereof.

7. A composition comprising a compound claim 1 or a salt thereof, and a thermostable DNA polymerase.

8. The composition of claim 7 , wherein the thermostable DNA polymerase is selected from the group consisting of: Taq, Tca, Tfu, Tbr, Tth, Tih, Tfi, Tli, Tfl, Pfu, Pwo, KOD, Tma, Tne, Bst, Pho, Sac, Sso, ES4, or a mutant, variant or derivative thereof.

9. The composition of claim 7 , wherein the compound is bound to the DNA polymerase and inhibits the activity of the DNA polymerase.

10. The composition of claim 7 , further comprising one or more nucleic acid amplification reagents selected from the group consisting of: deoxynucleotide triphosphates, buffer, a magnesium salt, an oligonucleotide primer, and a nucleic acid template.

11. A composition comprising a compound of claim 1 or a salt thereof, and one or more nucleic acid amplification reagents selected from the group consisting of: a polymerase, deoxynucleotide triphosphates, buffer, a magnesium salt, an oligonucleotide primer, and a nucleic acid template.

Assignees

• Promega Corp

Inventors

• Zhou Wenhui
• Knoche Kimberly K
• Storts Douglas R
• Zhou Min
• Meisenheimer Poncho

Classifications

• C07D307/80

  Radicals substituted by oxygen atoms · CPC title

• C07D307/56 Primary

  with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms · CPC title

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• C07C271/22

  to carbon atoms of hydrocarbon radicals substituted by carboxyl groups · CPC title

• C12Y207/07007

  DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase · CPC title