Chiral design

Patent metadata

Field	Value
Publication number	US-10160969-B2
Application number	US-201515112146-A
Country	US
Kind code	B2
Filing date	Jan 16, 2015
Priority date	Jan 16, 2014
Publication date	Dec 25, 2018
Grant date	Dec 25, 2018

Abstract

The present invention relates to chirally controlled oligonucleotides of select designs, chirally controlled oligonucleotide compositions, and methods of making and using the same. In some embodiments, a provided chirally controlled oligonucleotide composition provides different cleavage patterns of a nucleic acid polymer than a reference oligonucleotide composition. In some embodiments, a provided chirally controlled oligonucleotide composition provides single site cleavage within a complementary sequence of a nucleic acid polymer.

First claim

The invention claimed is:

1. A chirally controlled oligonucleotide composition comprising oligonucleotides of a particular oligonucleotide type characterized by:
1) a common base sequence and length;
2) a common pattern of backbone linkages; and
3) a common pattern of backbone chiral centers,

which composition is chirally controlled in that it is enriched, relative to a substantially racemic preparation of oligonucleotides having the same base sequence and length, for oligonucleotides of the particular oligonucleotide type; wherein:
the oligonucleotides of the particular oligonucleotide type each comprise three or more chiral, modified phosphate linkages;
the common pattern of backbone chiral centers comprises from 5′ to 3′ Rp(Sp) 2 ;
the common base sequence has at least 17 bases; and
the common pattern of backbone chiral centers comprises at least 50% of backbone chiral centers in the Sp conformation.

2. The composition of claim 1 , wherein each chiral, modified phosphate linkage of the oligonucleotides of the particular oligonucleotide type independently has the structure of formula I:

wherein:
P* is an asymmetric phosphorus atom and is either Rp or Sp;
W is O, S or Se;
each of X, Y and Z is independently —O—, —S—, —N(—L—R 1 )—, or L;
L is a covalent bond or an optionally substituted, linear or branched C1-C10 alkylene, wherein one or more methylene units of L are optionally and independently replaced by an optionally substituted C 1 -C 6 alkylene, C 1 -C 6 alkenylene, —C≡C—, —C(R′) 2 , —Cy—, —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)′, —N(R′)C(O)N(R′)—, —N(R′)C(O)—, —N(R′)C(O)O—, —OC(O)N(R′)—, —S(O)—, —S(O) 2 —, —S(O) 2 N(R′)—, —N(R′)S(O) 2 —, —SC(O)—, —C(O)S—, —OC(O)—, or —C(O)O—;
R 1 is halogen, R, or an optionally substituted C 1 -C 50 aliphatic wherein one or more methylene units are optionally and independently replaced by an optionally substituted C 1 -C 6 alkylene, C 1 -C 6 alkenylene, —C≡C—, —C(R′) 2 —, —Cy—, —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)—, —N(R′)C(O)O—, —OC(O)N(R′)—, —S(O)—, —S(O) 2 —, —S(O) 2 N(R′)—, —N(R′)S(O) 2 —, —SC(O)—, —C(O)S—, —OC(O)—, or —C(O)O—;

each R′ is independently —R, —C(O)R, —CO 2 R, or —SO 2 R, or:
two R′ on the same nitrogen are taken together with their intervening atoms to form an optionally substituted heterocyclic or heteroaryl ring, or
two R′ on the same carbon are taken together with their intervening atoms to form an optionally substituted aryl, carbocyclic, heterocyclic, or heteroaryl ring;

—Cy— is an optionally substituted bivalent ring selected from phenylene, carbocyclylene, arylene, heteroarylene, or heterocyclylene;
each R is independently hydrogen, or an optionally substituted group selected from C 1 -C 6 aliphatic, phenyl, carbocyclyl, aryl, heteroaryl, or heterocyclyl; and each — — independently represents a connection to a nucleoside.

3. The composition of claim 2 , wherein W is O, X is —S—, and Y and Z are —O—.

4. The composition of claim 2 , wherein X is —S— and —L—R 1 is not hydrogen.

5. The composition of claim 2 , wherein each chiral, modified phosphate linkage of the oligonucleotides of the particular oligonucleotide type is a phosphorothioate diester linkage.

6. The composition of claim 5 , wherein the common pattern of backbone chiral centers comprises from 5′ to 3′ SpSpRpSpSp.

7. The composition of claim 5 , wherein the pattern of backbone chiral centers comprises from 5′ to 3′ (Np)t(Rp)n(Sp)m, wherein t is 1, 2, 3, 4, 5, 6, 7 or 8, m is 2, 3, 4, 5, 6, 7 or 8, n is 1, and each Np is independent Rp or Sp.

8. The composition of claim 7 , wherein Np is Sp.

9. The composition of claim 7 , wherein t is greater than 5.

10. The composition of claim 9 , wherein the pattern of backbone chiral centers comprises (Sp) 2 Rp(Sp) 2 .

11. The composition of claim 5 , wherein the oligonucleotides of the particular oligonucleotide type each comprise one or more phosphate diester linkages.

12. The composition of claim 5 , wherein the nucleobases of the oligonucleotides of the particular oligonucleotide type are independently selected from adenine, thymine, cytosine, guanine, uracil and 5-methylcytosine.

13. The composition of claim 5 , wherein the oligonucleotides of the particular oligonucleotide type each comprise a modified sugar.

14. The composition of claim 13 , wherein the modified sugar comprises a 2′-modification.

15. The composition of claim 14 , wherein the 2′-modification is 2′ —OR 1 , wherein R 1 is optionally substituted C 1 -C 6 aliphatic.

16. The composition of claim 14 , wherein the 2′-modification is 2′ —OMe.

17. The composition of claim 14 , wherein the 2′-modification is 2′ —OCH 2 CH 2 OMe.

18. The composition of claim 5 , wherein the oligonucleotides of the particular oligonucleotide type each comprise a locked nucleic acid sugar.

19. The composition of claim 13 , wherein the modified sugar comprises a bivalent substituent —L—, wherein —L— is —O—CH 2 — between C 2 and C 4 of the sugar, wherein the —CH 2 — is optionally substituted.

20. The composition of claim 13 , wherein the modified sugar comprises a bivalent substituent —L—, wherein —L— is —O—CH 2 (Et)— between C 2 and C 4 of the sugar.

21. The composition of claim 1 , wherein the oligonucleotides of the particular oligonucleotide type each comprise at least one chiral, modified phosphate linkage having the structure of formula I:

wherein:
P* is an asymmetric phosphorus atom and is either Rp or Sp;
W is O, S or Se;
each of X, Y and Z is independently —O—, —S—, —N(—L—R 1 ), or L;
L is a covalent bond or an optionally substituted, linear or branched C 1 -C 10 alkylene, wherein one or more methylene units of L are optionally and independently replaced by an optionally substituted C 1 -C 6 alkylene, C 1 -C 6 alkenylene, —C≡C—, —C(R′) 2 —, —Cy—, —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)—, —N(R′)C(O)O—, —OC(O)N(R′)—, —S(O)—, —S(O) 2 —, —S(O) 2 N(R′)—, —N(R′S(O) 2 —, —SC(O)—, —C(O)S—, —OC(O)—, or —C(O)O—;
R 1 is halogen, R, or an optionally substituted C 1 -C 50 aliphatic wherein one or more methylene units are optionally and independently replaced by an optionally substituted C 1 -C 6 alkylene, C 1 -C 6 alkenylene, —C≡C—, —C(R′) 2 —, —Cy—, —O—, —S—, —S—S—, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)—, —N(R′)C(O)O—, —OC(O)N(R′)—, —S(O)—, —S(O) 2 —, —S(O) 2 N(R′)—, —N(R′)S(O) 2 —, —SC(O)—, —C(O)S—, —OC(O)—, or —C(O)O—;
each R′ is independently —R, —C(O)R, —CO 2 R, or —SO 2 R, or:
two R′ on the same nitrogen are taken together with their intervening atoms to form an optionally substituted heterocyclic or heteroaryl ring, or
two R′ on the same carbon are taken together with their intervening atoms to form an optionally substituted aryl, carbocyclic, heterocyclic, or heteroaryl ring;

—Cy— is an optionally substituted bivalent ring selected from phenylene, carbocyclylene, arylene, heteroarylene, or heterocyclylene;
each R is independently hydrogen, or an optionally substituted group selected from C 1 -C 6 aliphatic, phenyl, carbocyclyl, aryl, heteroaryl, or heterocyclyl; and

Assignees

• Wave Life Sciences Ltd

Inventors

• Meena
• Butler David
• Iwamoto Naoki
• Svrzikapa Nenad
• Verdine Gregory L
• Zlatev Ivan

Classifications

• C12N2310/31

  of the backbone · CPC title

• C12N15/87

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title

• C12N15/113 Primary

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N2310/315

  Phosphorothioates · CPC title

• C12N2320/30

  Special therapeutic applications · CPC title