Polymerase inhibitors and related compositions and methods
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.
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.