Antisense oligomers targeting PCSK9
The present invention relates to oligomeric compounds and conjugates thereof that target Proprotein Convertase Subtilisin/Kexin type 9 (PCSK9) PCSK9 mRNA in a cell, leading to reduced expression of PCSK9. Reduction of PCSK9 expression is beneficial for a range of medical disorders, such as hypercholesterolemia and related disorders.
1. An antisense oligonucleotide (ASO) comprising 10 to 16 nucleotides from the sequence aatgctacaaaaccca (SEQ ID NO:26) wherein:
(i) the ASO is a gapmer 10 to 25 contiguous nucleotides in length;
(ii) the ASO comprises at least one nucleotide analogue;
(iii) the ASO targets mRNA encoding PCSK9; and,
(iv) the ASO shows less kidney toxicity than an ASO of SEQ ID NO:1 as determined by a KIM-1 expression assay.
2. The ASO of claim 1 , wherein the at least one nucleotide analogue comprises an LNA unit.
3. The ASO of claim 2 , wherein the LNA unit is oxy-LNA, thio-LNA, amino-5 LNA, 5′-methyl-LNA, ENA, cET, cMOE or a combination thereof.
4. The ASO of claim 1 , wherein all the internucleoside linkages are phosphorothioate, and at least one of the phosphorothioate internucleoside linkages comprises a chiral center in the R conformation or in the S conformation.
5. An ASO conjugate comprising and ASO of claim 1 and at least one non-nucleotide or non-polynucleotide moiety covalently attached to the ASO directly or via a linker positioned between the contiguous ASO sequence and the non-nucleotide or non-polynucleotide moiety.
6. The ASO conjugate of claim 5 , wherein the non-nucleotide or non-polynucleotide moiety is a liver targeting moiety that is attached to the 5′-end or to the 3′-end of the ASO.
7. The ASO conjugate of claim 6 , wherein the liver targeting moiety comprises at least one asialoglycoprotein receptor targeting conjugate moiety.
8. The ASO conjugate of claim 7 , wherein the asialoglycoprotein receptor targeting conjugate moiety comprises a monovalent, divalent, trivalent, or tetravalent GalNAc cluster.
9. A pharmaceutical composition comprising a compound comprising an ASO of claim 1 and a pharmaceutically acceptable diluent, carrier, salt, or adjuvant.
10. A method of treating a disorder selected from the group consisting of atherosclerosis, hyperlipidemia, hypercholesterolemia, HDL/LDL cholesterol imbalance, coronary artery disease (CAD), or coronary heart disease (CHD) in a subject in need thereof, the method comprising administering comprising administering an effective amount of a compound comprising an ASO of claim 1 to the subject.
11. The method of claim 10 , wherein the dyslipidemia is familial hyperlipidemia (FCHL) or acquired hyperlipidemia.
12. The method of claim 10 , wherein the hypercholesterolemia is familiar hypercholesterolemia or statin resistant hypercholesterolemia.
13. The method of claim 10 , further comprising the administration of a therapeutic agent selected from the group consisting of a statin, a bile sequestering resin, nicotinic acid, a fibric acid derivative, probucol, neomycin, dextrothyroxine, a plant stanol ester, a cholesterol absorption inhibitor, implitapide, an inhibitor of bile acid transporters, a regulator of hepatic CYP7a, an estrogen replacement therapeutic, and an anti-inflammatory.
14. The method of claim 13 , wherein the statin is selected from the group consisting of lovastatin, cerivastatin, pravastatin, atorvastatin, simvastatin, rosuvastatin, and fluvastatin.
15. The method of claim 10 , wherein the antisense oligonucleotide conjugate is administered intravenously or subcutaneously.
16. The method of claim 10 , wherein the antisense oligonucleotide conjugate is administered as a single dose or as multiple doses.
17. An in vitro method of reducing expression levels and/or activity of PCSK9 in a cell comprising administering an effective amount of a compound comprising an ASO of claim 1 to the cell.
18. A method of reducing expression levels and/or activity of PCSK9 in a subject in need thereof comprising administering an effective amount of a compound comprising an ASO of claim 1 to the subject.
19. A method of reducing cholesterol levels in a subject in need thereof comprising administering to said subject an effective amount of a compound comprising an ASO of claim 1 .
20. A method of manufacturing a compound comprising an ASO of claim 1 , the method comprising chemically synthesizing the compound using sequential synthesis.
21. The method of claim 20 , wherein the sequential synthesis is solid phase oligonucleotide synthesis.