RNA interference mediated inhibition of prolyl hydroxylase domain 2 (PHD2) gene expression using short interfering nucleic acid (siNA)
View Patent ↗The present invention relates to compounds, compositions, and methods for the study, diagnosis, and treatment of traits, diseases and conditions that respond to the modulation of PHD2 gene expression and/or activity, and/or modulate a beta-catenin gene expression pathway. Specifically, the invention relates to double-stranded nucleic acid molecules including small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsR-NA), micro-RNA (miRNA), and short hair-pin RNA (shRNA) molecules that are capable of mediating or that mediate RNA interference (RNAi) against PHD2 gene expression.
1. An isolated double-stranded short interfering nucleic acid (siNA) molecule that inhibits expression of prolyl hydroxylase domain 2 (PHD2), comprising
5′-B c A uu GAA ccc AAA uuu GA u TT B-3′
(SEQ ID NO: 347)
and
5′-AUC AAA uuu GGG uuc AA u GUU -3′;
(SEQ ID NO: 348)
5′-B G c AA u AA cu G uuu GG u A uu TT B-3′
(SEQ ID NO: 312)
and
5′-AAUAcc AAA c AG uu A uu G c UU -3′;
(SEQ ID NO: 313)
5′-B G u GA c A u G u A u A u A uu A uc TT B-3′
(SEQ ID NO: 470)
and
5′-GAU AA u A u A u A c A u G uc A c UU -3′;
(SEQ ID NO: 471)
5′-B A u G cu A c AAGG u A c G c AA u TT B-3′
(SEQ ID NO: 405)
and
5′-AUU G c G u A ccuu G u AG c A u UU -3′;
(SEQ ID NO: 406)
5′-B AAGG u A c G c AA u AA cu G uu TT B-3′
(SEQ ID NO: 500)
and
5′-AAC AG uu A uu G c G u A ccuu UU -3′;
(SEQ ID NO: 501)
5′-Bcuu G uuu G u GG u A cuuc A u TT B-3′
(SEQ ID NO: 1055)
and
5′-AUG AAG u A cc A c A A A c AAGUU -3′;
(SEQ ID NO: 1056)
5′-B GAAAGG u G uuc AAG u A cc ATT B-3′
(SEQ ID NO: 1057)
and
5′-UGGu A cuu GAA c A ccuuuc UU -3′;
(SEQ ID NO: 1058)
or
5′-B ccu G u A ucu A cu A ccu GAATT B-3′
(SEQ ID NO: 1059)
and
5′-UUC AGG u AG u AGA u A c AGGUU -3′;
(SEQ ID NO: 1060)
wherein A, C, G, and U are ribose A, C, G or U; a, g, c and u are 2′-deoxy-2′-fluoro A, G, C or U; A, U, C and G are 2′-O-methyl (2′-OMe) A, U, C, or G; A, U, C, and G are deoxy A, U, C, or G; B is inverted abasic; and T is thymidine.
2. A composition comprising one or more siNA molecules of claim 1 and a pharmaceutically acceptable carrier or diluent.
3. A composition comprising:
(a) one or more siNA molecules of claim 1 ;
(b) a cationic lipid;
(c) cholesterol;
(d) DSPC; and
(e) PEG-DMG.
4. A composition comprising:
(a) one or more siNA molecules of claim 1 ;
(b) (13Z,16Z)—N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine;
(c) cholesterol;
(d) DSPC; and
(e) PEG-DMG.
5. The composition according to claim 4 , wherein the (13Z,16Z)—N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine, cholesterol, DSPC, and PEG-DMG have a molar ratio of 50:30:10:2 respectively.
6. A composition comprising the composition according to claim 5 and a pharmaceutically acceptable carrier or diluent.
7. A method of treating a human subject suffering from anemia, which comprises administering to said subject an effective amount of one or more siNA molecules of claim 1 , thereby treating the subject.
8. A kit comprising one or more siNA molecules of claim 1 .
9. A method for inhibiting the expression of prolyl hydroxylase domain 2 (PHD2) in a cell, comprising contacting the cell with a sufficient amount of one or more siNA molecules of claim 1 , thereby inhibiting expression of PHD2 in the cell.
10. A method for upregulating the expression of erythropoietin in a subject, comprising administering to the subject a sufficient amount of one or more siNA molecules of claim 1 , thereby upregulating the expression of erythropoietin in the subject.
11. The method of claim 10 , wherein the subject is a human subject.