Nucleic acid prodrugs and methods of use thereof

Patent metadata

Field	Value
Publication number	US-9744183-B2
Application number	US-201013381323-A
Country	US
Kind code	B2
Filing date	Jul 6, 2010
Priority date	Jul 6, 2009
Publication date	Aug 29, 2017
Grant date	Aug 29, 2017

Abstract

Described herein are nucleic acid prodrugs and nucleic acid prodrugs comprising chiral phosphorous moieties. Also described herein are methods of making and using nucleic acid prodrugs and nucleic acid prodrugs comprising chiral phosphorous moieties.

First claim

What is claimed is:

1. An oligonucleotide composition comprising a plurality of oligonucleotides of the following structure:

wherein:
each X-phosphonate independently has an Rp or Sp configuration;
R 1 is —OH, —SH, —NR d R d , —N 3 , halogen, hydrogen, alkyl, alkenyl, alkynyl, alkyl-Y 1 —, alkenyl-Y 1 —, alkynyl-Y 1 —, aryl-Y 1 —, heteroaryl-Y 1 —, —P(O)(R e ) 2 , —HP(O)(R e ), —OR a or —SR c ;
Y 1 is O, NR d , S, or Se;
R a is a blocking group;
R c is a blocking group;
each instance of R d is independently hydrogen, alkyl, alkenyl, alkynyl, aryl, acyl, substituted silyl, carbamate, —P(O)(R e ) 2 , or —HP(O)(R e );
each instance of R e is independently hydrogen, alkyl, aryl, alkenyl, alkynyl, alkyl-Y 2 —, alkenyl-Y 2 —, alkynyl-Y 2 —, aryl-Y 2 —, or heteroaryl-Y 2 —, or a cation which is Na +1 , Li +1 , or K +1 ;
Y 2 is O, S, or NR d wherein R d is hydrogen, alkyl, alkenyl, alkynyl, aryl, acyl, substituted silyl, or carbamate;
each instance of R 2 is independently hydrogen, —OH, —SH, —NR d R d , —N 3 , halogen, alkyl, alkenyl, alkynyl, alkyl-Y 1 —, alkenyl-Y 1 —, alkynyl-Y 1 —, aryl-Y 1 —, heteroaryl-Y 1 —, —OR b , or —SR c , wherein R b is a blocking group;
each instance of Ba is independently a blocked or unblocked adenine, cytosine, guanine, thymine, uracil or modified nucleobase, wherein each modified nucleobase is independently adenine, cytosine, guanine, thymine or uracil, modified by one or more modifications by which:
(1) a nucleobase is modified by one or more groups independently selected from acyl, halogen, amino, azide, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heterocyclyl, heteroaryl, carboxyl, hydroxyl, biotin, avidin, streptavidin, substituted silyl, and combinations thereof;
(2) one or more atoms of a nucleobase are independently replaced with a different atom selected from carbon, nitrogen or sulfur;
(3) one or more double bonds in a nucleobase are independently hydrogenated; or
(4) one or more aryl or heteroaryl rings are independently inserted into a nucleobase; or is selected from:
uracil, thymine, adenine, cytosine, and guanine having their respective amino groups protected by acyl groups, 2-fluorouracil, 2-fluorocytosine, 5-bromouracil, 5-iodouracil, 2,6-diaminopurine, azacytosine, pseudoisocytosine, pseudouracil, 8-substituted purines, xanthine, hypoxanthine,

wherein:
R 8 is a linear or branched alkyl, aryl, aralkyl, or aryloxylalkyl group having 1 to 15 carbon atoms, and
each of R 9 and R 10 represents a linear or branched alkyl group having 1 to 4 carbon atoms;

corrin or porphyrin, each optionally modified by one or more groups independently selected from acyl, halogen, amino, azide, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heterocyclyl, heteroaryl, carboxyl, hydroxyl, biotin, avidin, streptavidin, substituted silyl, and combinations thereof;
phenanthrene, pyrene, stillbene, isoxanthine, isozanthopterin, terphenyl, terthiophene, benzoterthiophene, coumarin, lumazine, tethered stillbene, benzo-uracil, or naphtho-uracil, each optionally modified by one or more groups independently selected from acyl, halogen, amino, azide, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heterocyclyl, heteroaryl, carboxyl, hydroxyl, biotin, avidin, streptavidin, substituted silyl, and combinations thereof;
3-nitropyrrole, 5-bromouracil, 5-iodouracil, or 2,6-diaminopurine, or a nucleobase of 4-acetylcytidine; 5-(carboxyhydroxylmethyl)uridine; 2′-O-methylcytidine; 5-carboxymethylaminomethyl-2-thiouridine; 5-carboxymethylaminomethyluridine; dihydrouridine; 2′-O-methylpseudouridine; beta,D-galactosylqueosine; 2′-O-methylguanosine; N6-isopentenyladenosine; 1-methyladenosine; 1-methylpseudouridine; 1-methylguanosine; 1-methylinosine; 2,2-dimethylguanosine; 2-methyladenosine; 2-methylguanosine; N7-methylguanosine; 3-methyl-cytidine; 5-methylcytidine; N6-methyladenosine; 7-methylguanosine; 5-methylaminoethyluridine; 5-methoxyaminomethyl-2-thiouridine; beta,D-mannosylqueosine; 5-methoxycarbonylmethyluridine; 5-methoxyuridine; 2-methylthio-N6-isopentenyladenosine; N-((9-beta,D-ribofuranosyl-2-methylthiopurine-6-yl)carbamoyl)threonine; N-((9-beta,D-ribofuranosylpurine-6-yl)-N-methylcarbamoyl)threonine; uridine-5-oxyacetic acid methylester; uridine-5-oxyacetic acid (v); pseudouridine; queosine; 2-thiocytidine; 5-methyl-2-thiouridine; 2-thiouridine; 4-thiouridine; 5-methyluridine; 2′-O-methyl-5-methyluridine; or 2′-O-methyluridine; and
heteroaryl or heterocyclyl optionally substituted with one or more groups independently selected from acyl, halogen, amino, azide, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heterocyclyl, heteroaryl, carboxyl, hydroxyl, biotin, avidin, streptavidin, substituted silyl, and combinations thereof;
each instance of X is H or —OCH 2 CH 2 S—S(O) 2 R 10 , —OCH 2 CH 2 S—SCH 2 CH 2 OH, —OCH 2 CH 2 CO 2 H,

wherein at least 25% of X moieties present in the oligonucleotide are not H;
wherein R 10 is an alkyl group having 1 to 4 carbon atoms; R 11 is alkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl; and R 12 is hydrogen or alkyl;
R 3 is hydrogen, a blocking group, a linking moiety connected to a solid support or a linking moiety connected to a nucleic acid;
n is an integer of 10 to about 200; and
wherein the composition is stereodefined in that each X-phosphonate moiety is more than 98% diastereomerically pure within the composition.

2. The composition of claim 1 wherein the diastereomeric purity is determined by 31 P NMR spectroscopy or reverse-phase HPLC.

3. The composition of claim 1 wherein each X-phosphonate moiety has an R P configuration.

4. The composition of claim 1 wherein each X-phosphonate moiety has an S P configuration.

5. The composition of claim 1 , wherein at least 90% of the X moieties of the oligonucleotide are independently selected from —OCH 2 CH 2 S—S(O) 2 R 10 , —OCH 2 CH 2 S—SCH 2 CH 2 OH, —OCH 2 CH 2 CO 2 H,

wherein R 10 is an alkyl group having 1 to 4 carbon atoms; R 11 is alkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl; and R 12 is hydrogen or alkyl.

6. The composition of claim 1 , wherein each instance of X is independently selected from —OCH 2 CH 2 S—S(O) 2 R 10 , —OCH 2 CH 2 S—SCH 2 CH 2 OH, —OCH 2 CH 2 CO 2 H,

wherein R 10 is an alkyl group having 1 to 4 carbon atoms; R 11 is alkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl; and R 12 is hydrogen or alkyl.

7. The composition of claim 1 , wherein R 10 is methyl.

8. The composition of claim 1 , wherein R 11 is methyl.

9. The composition of claim 1 , wherein R 12 is methyl.

10. The composition of claim 1 , wherein n is an integer of 15 to about 200.

11. The composition of claim 1 , wherein n is an integer of 2

Assignees

• Verdine Gregory L
• Meena
• Iwamoto Naoki
• Wave Life Sciences Ltd

Inventors

• Verdine Gregory L
• Meena
• Iwamoto Naoki

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P3/02

  Nutrients, e.g. vitamins, minerals · CPC title

• A61P1/18

  for pancreatic disorders, e.g. pancreatic enzymes · CPC title

• A61K31/70 Primary

  Carbohydrates; Sugars; Derivatives thereof (sorbitol A61K31/047) · CPC title