Oligomeric compounds for the modulation of HIF-1A expression
View Patent ↗Oligonucleotides directed against the hypoxia-inducible factor-1α (HIF-1α) gene are provided for modulating the expression of HIF-1α. The compositions comprise oligonucleotides, particularly antisense oligonucleotides, targeted to nucleic acids encoding the HIF-1α. Methods of using these compounds for modulation of HIF-1α expression and for the treatment of diseases associated with the hypoxia-inducible factor-1α are provided. Examples of diseases are cancer and pre-eclampsia. The oligonucleotides may be composed of deoxyribonucleosides, a nucleic acid analog, or Locked Nucleic Acid (LNA) or a combination thereof.
1. A compound of 16 to 50 nucleotides or nucleotide analogues, comprising SEQ ID NO: 3, wherein adjacent nucleotides or nucleotide analogues are covalently linked by an internucleotide linkage, wherein the compound comprises at least one nucleotide analogue, and wherein the compound modulates HIF-1α expression.
2. The compound according to claim 1 which inhibits the expression of HIF-1α in a cell or tissue.
3. The compound according to claim 1 , which is an antisense oligonucleotide.
4. The compound according to claim 3 , wherein the nucleotide analogue is a Locked Nucleic Acid (LNA) unit.
5. The compound according to claim 4 , wherein the LNA unit has the structure:
wherein Z and Z* are independently selected from the group consisting of an internucleotide linkage, a terminal group and a protecting group,
B constitutes a natural or non-natural nucleobase,
X and Y are independently selected from —O—, —S—, —N(H)—, —N(R)—, —CH 2 —, —CH— (if part of a double bond), —CH 2 —O—, —CH 2 —S—, —CH 2 —N(H)—, —CH 2 —N(R)—, —CH 2 —CH 2 —, and
—CH 2 —CH— (if part of a double bond) in which R H is selected from hydrogen and C 1-4 -alkyl.
6. The compound according to claim 4 , wherein the LNA unit has a structure of any of the following formulas:
wherein X is independently selected from —O—, —S—, —NH—, and N(R H ) in which R H is selected from hydrogen and C 1-4 -alkyl,
Z and Z* are independently selected from the group consisting of an internucleotide linkage, a terminal group and a protecting group; and
B constitutes a natural or non-natural nucleobase.
7. The compound according to claim 5 , wherein the internucleotide linkage is selected from the group consisting of —O—P(O) 2 —O—, —O—P(O,S)—O—, —O—P(S) 2 —O—, —S—P(O) 2 —O—, —S—P(O,S)—O—, —S—P(S) 2 —O—, —O—P(O) 2 —S—, —O—P(O,S)—S—, —S—P(O) 2 —S—, —O—PO(R H )—O—, O—PO(OCH 3 )—O—, —O—PO(NR H )—O—, —O—PO(OCH 2 CH 2 S—R)—O—, —O—PO(BH 3 )—O—, —O—PO(NHR H )—O—, —O—P(O) 2 —HR H —, —NR H —P(O) 2 —O—, and —NR″—CO—O—, where R H is selected form hydrogen and C 1-4 -alkyl.
8. The compound of claim 7 , wherein at least one internucleotide linkage is a phosphorothioate linkage.
9. The compound according to claim 5 , wherein at least one nucleobase is a modified nucleobase independently selected from the group consisting of 5-methylcytosine, isocytosine, pseudoisocytosine, 5-bromouracil, 5-propynyluracil, 6-aminopurine, 2-aminopurine, inosine, diaminopurine, and 2-chloro-6-aminopurine.
10. The compound according to claim 4 , wherein each of the LNA units is independently selected from β-D-oxy-LNA, β-D-thio-LNA, β-D-amino-LNA, α-L-oxy-LNA, α-L-thio-LNA and α-L-amino-LNA.
11. The compound of claim 3 , wherein the antisense oligonucleotide is a chimeric oligonucleotide.
12. The compound of claim 11 , wherein the chimeric oligonucleotide is a gapmer, a headmer, or a tailmer.
13. The compound according to claim 4 , which comprises at least four LNA units.
14. A composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier or diluent.
15. A pharmaceutical composition comprising the compound of claim 1 in the form of a pharmaceutically acceptable salt.
16. A pharmaceutical composition comprising the compound of claim 1 , which is in the form of a conjugate.
17. A pharmaceutical composition comprising the compound of claim 1 , which is in the form of a pro-drug.
18. A compound represented by the formula: T s G s G s T s g s a s g s g s c s t s g s t s C s C s G s A (SEQ ID NO. 3) wherein uppercase letters denote oxy-LNA nucleotides, lowercase letters denote deoxynucleotides and the subscript “s” denotes a phosphorothioate linkage.
19. A compound selected from:
(SEQ ID NO. 3)
T s G s G s T s g s a s g s g s c s t s g s t s Me C s Me C s G s A,
(SEQ ID NO. 96)
t s g s g s t s g s a s g s g s c s t s g s t s c s c s g s a,
(SEQ ID NO. 97)
TGGT s g s a s g s g s c s t s g s t s Me C Me CGA,
(SEQ ID NO. 99)
TGGTGAGG s c s t s g s t s c s c s g s a
(SEQ ID NO. 100)
t s g s g s t s g s a s g s g s Me CTGT Me C Me CGA
(SEQ ID NO. 105)
T s G s G s t s g s a s g s g s c s t s g s t s Me C s Me C s G s A,
(SEQ ID NO. 106)
T s G s G s t s g s a s g s g s c s t s g s t s Me C s Me C s G s a,
(SEQ ID NO. 107)
T s G s G s T s g s a s g s g s c s t s g s t s Me C s Me C s G s a,
(SEQ ID NO. 108)
T s G s G s T s G s A s g s g s c s t s g s t s c s c s g s a,
(SEQ ID NO. 109)
TGGt s g s a s g s g s c s t s g s t s Me C Me CGA,
(SEQ ID NO. 110)
TGGt s g s a s g s g s c s t s g s t s Me C Me CGa,
(SEQ ID NO. 111)
TGGTg s a s g s g s c s t s g s t s Me C Me CGa,
and
(SEQ ID NO. 112)
TGGTGAg s g s c s t s g s t s c s c s g s a,
wherein uppercase letters denote oxy-LNA nucleotides, lowercase letters denote deoxynucleotides, the subscript “s” denotes a phosphorothioate linkage, and wherein Me C denotes an oxy LNA monomer containing a 5-methyl cytosine base.