Trinucleotide mRNA cap analogs

Patent metadata

Field	Value
Publication number	US-11866754-B2
Application number	US-201615768206-A
Country	US
Kind code	B2
Filing date	Oct 17, 2016
Priority date	Oct 16, 2015
Publication date	Jan 9, 2024
Grant date	Jan 9, 2024

Abstract

The present disclosure relates to cap analogs, which can result in high levels of capping efficiency and transcription and improved translation efficiencies. The present disclosure also relates to methods useful for preparing cap analogs and using mRNA species containing such analogs, as well as kits containing the novel cap analogs.

First claim

What is claimed is:

1. A cap analog of formula (I):

or a stereoisomer, tautomer or salt thereof, wherein

is

ring B 1 is

 in which
R 1 is C 1 alkyl;
each of R a and R b is H; and
R c is H;
ring B 2 and ring B 3 are each independently selected from

wherein
X 1 is N or N + (R 5 );
R 5 is C 1 -C 6 alkyl, C 2 -C 6 alkenyl, or C 2 -C 6 alkynyl, each of which is optionally substituted with one or more substituents selected from the group consisting of C 6 -C 10 aryl, C 6 -C 10 aryloxyl, 5- to 10-membered heteroaryl, and 5- to 10-membered heteroaryloxyl, each being optionally substituted with one or more of halo and cyano;
each of R d and R e independently is H, C 1 -C 6 alkyl, or an amine protecting group, or R d and R e , together with the nitrogen atom to which they attach, form a 4 to 12-membered heterocycloalkyl, —N═CH—Ru, or —N═N—Ru, wherein R B is phenyl and each of the 4 to 12-membered heterocycloalkyl and R B is optionally substituted with one or more substituents selected from OH, halo, oxo, C 1 -C 6 alkyl, COOH, C(O)O—C 1 -C 6 alkyl, cyano, C 1 -C 6 alkoxyl, amino, mono-C 1 -C 6 alkylamino, and di-C 1 -C 6 alkylamino;
R f is H, NH 2 , or C 1 -C 6 alkyl; or R f and one of R d and R e , together with the two nitrogen atoms to which they attach and the carbon atom connecting the two nitrogen atoms form a 5- or 6-membered heterocycle which is optionally substituted with one or more of OH, halo, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, and C 2 -C 6 alkynyl;
R g is H or methyl;
R h is H or methyl;
X 2 is O;
Y 2 is (OP(O)R 4 ) m in which m is 1;
R 2 is locked nucleic acid (LNA);
R 2′ is OR 3 ;
R 3 is H;
each of R 4 and R 4′ is OH;
each of R 20 , R 21 , R 22 , and R 23 is —Q 3 -T 3 , in which Q 3 is a bond, and T 3 is H;
each of R 27 and R 28 is OR 29 ; and
each R 29 is H.

2. The cap analog of claim 1 , wherein the cap analog has a residence time of about 10 seconds or longer when binding with the eukaryotic initiation factor 4E (eIF4E) characterized by surface plasmon resonance (SPR).

3. A kit for capping an RNA transcript comprising a cap analog of claim 1 , and an RNA polymerase.

4. A method for synthesizing an RNA molecule whose 5′ end comprises a cap analog of claim 1 in vitro, the method comprising reacting unmodified or modified ATP, unmodified or modified CTP, unmodified or modified UTP, unmodified or modified GTP, the cap analog or a stereoisomer or salt thereof, and a polynucleotide template; in the presence an RNA polymerase; under a condition conducive to transcription by the RNA polymerase of the polynucleotide template into one or more RNA copies; whereby at least some of the RNA copies incorporate the cap analog or a stereoisomer or salt thereof to make the RNA molecule.

5. The cap analog of claim 1 , wherein ring Ba is

6. The cap analog of claim 1 , wherein ring B 2 is

7. The cap analog of claim 1 , wherein ring B 2 is

8. The cap analog of claim 1 , wherein ring B 2 is

9. The cap analog of claim 1 , wherein ring B 2 is

10. The cap analog of claim 1 , wherein ring B 2 is

11. The cap analog of claim 1 , wherein ring B 3 is

12. The cap analog of claim 1 , wherein ring B 3 is

13. The cap analog of claim 1 , wherein ring B 3 is

14. The cap analog of claim 1 , wherein ring B 3 is

15. The cap analog of claim 1 , wherein ring B 3 is

16. The cap analog of claim 1 , wherein ring B 3 is

17. The cap analog of claim 1 , wherein the cap analog has the following structure:

18. The cap analog of claim 5 , wherein ring B 3 is

Assignees

• Modernatx Inc

Inventors

• Butora Gabor
• Stanton Matthew
• Miracco Edward John

Classifications

• C12P19/34 Primary

  Polynucleotides, e.g. nucleic acids, oligoribonucleotides · CPC title

• A61K31/7125

  Nucleic acids or oligonucleotides having modified internucleoside linkage, i.e. other than 3'-5' phosphodiesters · CPC title

• C07H21/02 Primary

  with ribosyl as saccharide radical · CPC title

• C12N9/1247

  DNA-directed RNA polymerase (2.7.7.6) · CPC title

• C12N15/1096

  cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR · CPC title