US 11,987,599 B2
Solid support for synthesizing nucleic acid sequences and methods for making and using
Serge L. Beaucage, Silver Spring, MD (US); and Andrzej M. Grajkowski, Kensington, MD (US)
Assigned to United States of America, as represented by the Secretary, Department of Health and Human Services, Silver Spring, MD (US)
Appl. No. 18/003,404
Filed by United States of America, as represented by the Secretary, Department of Health and Human Services, Silver Spring, MD (US)
PCT Filed Jun. 28, 2021, PCT No. PCT/US2021/039403
§ 371(c)(1), (2) Date Dec. 27, 2022,
PCT Pub. No. WO2022/005988, PCT Pub. Date Jan. 6, 2022.
Claims priority of provisional application 63/046,413, filed on Jun. 30, 2020.
Prior Publication US 2023/0271998 A1, Aug. 31, 2023
Int. Cl. C07H 21/04 (2006.01); C07F 9/09 (2006.01); C07F 9/655 (2006.01); C07H 21/00 (2006.01); C07H 21/02 (2006.01); C12N 15/10 (2006.01); C12N 15/113 (2010.01); C40B 50/18 (2006.01)
CPC C07H 21/04 (2013.01) [C07F 9/093 (2013.01); C07F 9/098 (2013.01); C07F 9/65515 (2013.01); C07H 21/00 (2013.01); C07H 21/02 (2013.01); C12N 15/1068 (2013.01); C12N 15/113 (2013.01); C40B 50/18 (2013.01)] 33 Claims
 
1. A solid support according to Formula I

OG Complex Work Unit Chemistry
wherein:
CPG is controlled pore glass;
m is 2, 3, 4, 5 or 6;
x is 1, 2, 3, 4 or 5;
y is from 2 to 12;
n is 3, 4, 5, 6, 7, 8, 9 or 10;
R2 is H,

OG Complex Work Unit Chemistry
p is 2, 3, 4, 5, 6, 7, 8, 9 or 10;
R3 is H or

OG Complex Work Unit Chemistry
R4 is H or OR6;
R is PG or a nucleic acid sequence;
R6 is pixyl, TBDMS, TBDPS, TMS, TES, or TIPS;
T is 1, 2, 3, or 4;
BP is a nucleic acid base where an exocyclic amine, if present, is protected;
PG is a protecting group; and
each R1 independently is a thermolabile phosphate protecting group having a structure

OG Complex Work Unit Chemistry
wherein:
X is O or S;
R7 is H, Ra, ORa, SRa, or N(Rb)2;
Ra is Rd;
Rb is H, Rd or two Rbs together with the nitrogen to which they are attached, form a 3- to 7-membered heterocyclyl;
Z is O, S, N(Rc), C(Rc)2 or C(Rc)2C(Rc)2;
each Rc independently is H or Rd, or one Rd in combination with the C═X moiety and one Ra or Rb from R7 together form a 3- to 7-membered cycloaliphatic or heterocyclyl ring;
Rd is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or aralkyl;
each R8 independently is H or Rd, or one R8 together with Z forms an aryl ring;
each R9 independently is H or Rd, or one R9 and one R8 together with the atoms to which they are attached, forms a moiety having a formula

OG Complex Work Unit Chemistry
wherein r is 0 to 6; and
each R10 independently is H, C1-6alkyl, NO2, —N(C1-6alkyl)2, —OC1-6alkyl, —SC1-6alkyl, —CN, or halogen, provided that the aromatic ring substituted with R10 is one carbon removed from the phosphate oxygen of Formula I.