Oligonucleotide compounds for treatment of preeclampsia and other angiogenic disorders

Patent metadata

Field	Value
Publication number	US-9862952-B2
Application number	US-201615089437-A
Country	US
Kind code	B2
Filing date	Apr 1, 2016
Priority date	Apr 3, 2015
Publication date	Jan 9, 2018
Grant date	Jan 9, 2018

Abstract

This disclosure relates to novel targets for angiogenic disorders. Novel oligonucleotides are also provided. Methods of using the novel oligonucleotides for the treatment of angiogenic disorders (e.g., preeclampsia) are also provided.

First claim

What is claimed:

1. A compound, comprising a double stranded (ds) RNA molecule that is between 15 and 35 bases in length having a 5′ end, a 3′ end and complementarity to a target, and comprising a first oligonucleotide and a second oligonucleotide, that binds to an intronic region of an mRNA encoding an sFLT1 protein, wherein:
(1) a portion of the first oligonucleotide is complementary to a portion of the second oligonucleotide;
(2) the second oligonucleotide comprises alternating 2′-methoxy-ribonucleotides and 2′-fluoro-ribonucleotides;
(3) the nucleotides at positions 2 and 14 from the 3′ end of the second oligonucleotide are 2′-methoxy-ribonucleotides;
(4) the nucleotides of the second oligonucleotide are connected via phosphodiester or phosphorothioate linkages; and
(5) the compound comprises a hydrophobic modification,

wherein the compound selectively inhibits expression of the sFLT1 protein in a cell or organism.

2. A compound comprising a dsRNA that binds to an intronic region of an mRNA encoding an sFLT1 protein, said dsRNA is between 15 and 35 bases in length having a 5′ end, a 3′ end and complementarity to a target, and comprising a first oligonucleotide and a second oligonucleotide, wherein:
(1) the first oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4;
(2) a portion of the first oligonucleotide is complementary to a portion of the second oligonucleotide;
(3) the second oligonucleotide comprises alternating 2′-methoxy-ribonucleotides and 2′-fluoro-ribonucleotides;
(4) the nucleotides at positions 2 and 14 from the 3′ end of the second oligonucleotide are 2′-methoxy-ribonucleotides; and
(5) the nucleotides of the second oligonucleotide are connected via phosphodiester or phosphorothioate linkages; and
(6) the compound comprises a hydrophobic modification,

wherein the compound selectively inhibits expression of the sFLT1 protein in a cell or organism.

3. The compound of claim 2 , comprising a dsRNA having a sense strand and an antisense strand, wherein the antisense strand comprises a region of complementarity which is substantially complementary to
SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4, or
contains no more than 3 mismatches with SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4.

4. The dsRNA of claim 3 , wherein said region of complementarity:
is complementary to at least 15, 16, 17 or 18 contiguous nucleotides of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4;
contains no more than 3 mismatches with SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4; or
is fully complementary to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4.

5. The dsRNA of claim 3 , wherein said dsRNA: is blunt-ended and optionally;
comprises at least one single stranded nucleotide overhang; or
comprises naturally occurring nucleotides.

6. The dsRNA of claim 5 , wherein:
said modified nucleotide is chosen from the group consisting of a a terminal nucleotide linked to a cholesteryl derivative, a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.

7. A therapeutic composition that binds to an intronic region of an mRNA encoding an sFLT1 protein, wherein the therapeutic composition selectively reduces expression of the sFLT1 protein, and wherein the therapeutic composition reduces one or more symptoms of preeclampsia (PE), postpartum PE, eclampsia or Hemolysis/Elevated Liver enzymes/Low Platelet count (HELLP) syndrome when administered to a subject in need thereof, comprising a first dsRNA comprising a first sense strand and a first antisense strand and a second dsRNA comprising a second sense strand and a second antisense strand, wherein the first antisense strand comprises a first region of complementarity which is substantially complementary to SEQ ID NO:1 and the second antisense strand comprises a second region of complementarity which is substantially complementary to SEQ ID NO:2.

8. The therapeutic composition of claim 7 , wherein each dsRNA comprises at least one modified nucleotide.

9. The composition of claim 7 , wherein each dsRNA comprises a 5′ end, a 3′ end and complementarity to a target, wherein:
(1) the oligonucleotide comprises alternating 2′-methoxy-ribonucleotides and 2′-fluoro-ribonucleotides;
(2) the nucleotides at positions 2 and 14 from the 5′ end are not 2′-methoxy-ribonucleotides;
(3) the nucleotides are connected via phosphodiester or phosphorothioate linkages; and
(4) the nucleotides at positions 1-6 from the 3′ end, or positions 1-7 from the 3′ end, are connected to adjacent nucleotides via phosphorothioate linkages.

10. A pharmaceutical composition comprising:
a first dsRNA comprising a first sense strand and a first antisense strand, wherein the first antisense strand comprises a region of complementarity which is substantially complementary to SEQ ID NO:1, and wherein the first antisense strand selectively targets one or both of an intronic region of sFLT-i13 short and an intronic region of sFLT-i13 long;
a second dsRNA comprising a second sense strand and a second antisense strand, wherein the second antisense strand comprises a region of complementarity which is substantially complementary to SEQ ID NO:2, and wherein the second antisense strand selectively targets an intronic region of sFLT-i15a; and
a pharmaceutically acceptable carrier.

11. A method of treating or managing PE, postpartum PE, eclampsia or HELLP syndrome comprising administering to a subject in need of such treatment or management a therapeutically effective amount of the pharmaceutical composition of claim 10 .

12. The method of claim 11 , wherein the pharmaceutical composition is administered intravenously or subcutaneously.

13. A method of treating one or more symptoms of PE, postpartum PE, eclampsia or HELLP syndrome in a subject in need thereof, comprising administering to the subject the therapeutic compound of claim 7 .

14. The dsRNA of claim 2 , wherein:
the hydrophobic molecule comprises an omega-3 fatty acid; or
the hydrophobic molecule comprises a molecule selected from the group consisting of: docosanoic acid (DCA), docosahexaenoic acid (DHA), lysophosphatidylcholine esterified DHA (g2-DHA) and eicosapentaenoic acid (EPA).

15. The dsRNA of claim 14 , wherein the hydrophobic molecule is DHA or g2-DHA.

16. A compound comprising a dsRNA having a sense strand and an antisense strand, wherein the antisense strand comprises a region of complementarity which is substantially complementary to
(SEQ ID NO:1),
(SEQ ID NO:2),
(SEQ ID NO:3),
(SEQ ID NO:4), or
contains no more than 3 mismatches with SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4,
wherein said dsRNA comprises at least one 2′-O-methyl modified nucleotide, at least one 2′-fluoro modified nucleotide, at least one nucleotide comprising a 5′phosphorothioate group and wherein said dsRNA comprises a hydrophobic modification.

17. A compound comprising a dsRNA having a sense strand and an antisense strand, wherein the antisense strand comprises a region of complementarity which is substantially complementary to
(SEQ ID NO:1),
(SEQ ID NO:2),
(SEQ ID NO:3),
(SEQ ID NO:4), or
contains no more than 3 mismatches with SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or SEQ ID NO:4,
wherein said dsRNA comprises at least one 2′-O-methyl modified nucleotide, at

Assignees

• Univ Massachusetts
• Beth Israel Deaconess Medical Ct Inc
• Beth Israel Deaconess Medical Ct Inc

Inventors

• Khvorova Anastasia
• Moore Melissa
• Turanov Anton A
• Karumanchi Ananth

Classifications

• C12N2310/322

  2'-R Modification · CPC title

• C12N2310/344

  Position-specific modifications, e.g. on every purine, at the 3'-end · CPC title

• C12N2310/321

  2'-O-R Modification · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• A61K31/713

  Double-stranded nucleic acids or oligonucleotides · CPC title