US 12,337,009 B2
Nucleotide derivatives and methods of use thereof
Jingyue Ju, Englewwod Cliffs, NJ (US); Xin Chen, Changsha (CN); Xiaoxu Li, New York, NY (US); Zengmin Li, Flushing, NY (US); Min-Kang Hsieh, Radford, VA (US); Minchen Chien, Tenafly, NJ (US); Shundi Shi, Ozone Park, NY (US); Jianyi Ren, Rosemead, CA (US); Cheng Guo, Brooklyn, NY (US); Shiv Kumar, Belle Mead, NJ (US); James Russo, New York, NY (US); Chuanjuan Tao, Fort Lee, NJ (US); Steffen Jockusch, New York, NY (US); and Sergey Kalachikov, Bronx, NY (US)
Assigned to THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK, New York, NY (US)
Filed by The Trustees of Columbia University in the City of New York, New York, NY (US)
Filed on Mar. 7, 2022, as Appl. No. 17/688,752.
Application 17/688,752 is a division of application No. 16/303,715, granted, now 11,266,673, issued on Mar. 8, 2022, previously published as PCT/US2017/033939, filed on May 23, 2017.
Claims priority of provisional application 62/477,945, filed on Mar. 28, 2017.
Claims priority of provisional application 62/365,321, filed on Jul. 21, 2016.
Claims priority of provisional application 62/340,419, filed on May 23, 2016.
Prior Publication US 2023/0028321 A1, Jan. 26, 2023
Int. Cl. C07H 19/20 (2006.01); A61K 31/7052 (2006.01); C07H 21/04 (2006.01); C12Q 1/6869 (2018.01)
CPC A61K 31/7052 (2013.01) [C07H 21/04 (2013.01); C07H 19/20 (2013.01); C12Q 1/6869 (2013.01)] 20 Claims
OG exemplary drawing
 
1. A thermophilic nucleic acid polymerase complex comprising a thermophilic nucleic acid polymerase bound to a nucleotide analogue having the formula:

OG Complex Work Unit Chemistry
wherein
the symbol “----” is a non-covalent bond;
B is a base or analogue thereof;
L1 is a substituted or unsubstituted methylene, and wherein if L1 is a substituted methylene, L1 is substituted with a substituted or unsubstituted C1-C6 alkylene, substituted or unsubstituted 2 to 6 membered heteroalkylene, substituted or unsubstituted C3-C6 cycloalkylene, substituted or unsubstituted 3 to 6 membered heterocycloalkylene, substituted or unsubstituted phenyl, or substituted or unsubstituted 5 to 6 membered heteroarylene;
L2 is a cleavable linker or a non-cleavable linker;
R3 is triphosphate;
R4A is hydrogen, —CF3, —CCl3, —CBr3, —CI3, —CHF2, —CHCl2, —CHBr2, —CHI2, —CH2F, —CH2Cl, —CH2Br, —CH2I, —CN, substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl;
R4B is hydrogen, —CF3, —CCl3, —CBr3, —CI3, —CHF2, —CHCl2, —CHBr2, —CHI2, —CH2F, —CH2Cl, —CH2Br, —CH2I, —CN, —X—R6, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
wherein R6 is —CF3, —CCl3, —CBr3, —CI3, —CHF2, —CHCl2, —CHBr2, —CHI2, —CH2F, —CH2Cl, —CH2Br, —CH2I, —CN, substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl; and
wherein X is a bond, O, NR6A, or S;
wherein R6A is —OH, —CF3, —CCl3, —CBr3, —CI3, —CHF2, —CHCl2, —CHBr2, —CHI2, —CH2F, —CH2Cl, —CH2Br, —CH2I, —CN, substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl;
R5 is a detectable label or a biotin moiety;
R7 is hydrogen;
R12 is a streptavidin moiety;
L4 is an orthogonally cleavable linker; and
R13 is a fluorescent dye.