| PATENT ASSIGNEE'S COUNTRY | USA |
| UPDATE | 04.00 |
| PATENT NUMBER | This data is not available for free |
| PATENT GRANT DATE | 04.04.00 |
| PATENT TITLE |
Antisense modulation of G-alpha-i1 expression |
| PATENT ABSTRACT |
Antisense compounds, compositions and methods are provided for modulating the expression of G-alpha-i1. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding G-alpha-i1. Methods of using these compounds for modulation of G-alpha-i1 expression and for treatment of diseases associated with expression of G-alpha-i1 are provided. |
| PATENT INVENTORS | This data is not available for free |
| PATENT ASSIGNEE | This data is not available for free |
| PATENT FILE DATE | 09.04.99 |
| PATENT REFERENCES CITED |
Andrea D. Branch, A good antisense molecule is hard to find, TIBS, 47-48, Feb. 1998. Trisha Gura, Antisense Has Growing Pains, Science, pp. 575-577, Oct. 1995. Stanley Crooke, Antisense '97: A roundtable on the state of the industry, Nature Biotechnology, p. 522, Jun. 1997. Stanley Crooke, Antisense Research and Applications, Chapter 1, Basic Principles of Antisense Therapeutics, Springer-Verlag Press, Berlin, Heidelberg, New York, p. 3, Jul. 1998. Hamm, The many faces of G protein signaling, J. Biol. Chem., 1998, 273:669-672. Redmond, Flow-mediated regulation of G-protein expression in cocultured vascular smooth muscle and endothelial cells, Arterioscler. Thromb. Vasc. Biol., 1998, 18:75-83. Schmidt, Alterations in guanine nucleotide regulatory protein expression and activity in human hepatocellular carcinoma, Hepatology, 1997, 26:1189-1194. Svoboda, Cold-induced reduction in Gi alpha proteins in brown adipose tissue. Effects on the cellular hypersensitization to noradrenaline caused by pertussin-toxin treatment, Biochem. J., 1996, 314:761-768. Wise, Interactions of the alpha2A-adrenoceptor with multiple Gi-family G- proteins: studies with pertussis toxin-resistant G-protein mutants [published erratum appears in Biochem J 1997 May 1;323 (Pt 3):864], Biochem. J., 1997, 321:721-728. |
| PATENT CLAIMS |
What is claimed is: 1. An antisense compound 8 to 30 nucleobases in length targeted to a nucleic acid molecule encoding human G-alpha-i1, excluding a region complementary to SEQ ID NO: 15, wherein said antisense compound inhibits the expression of human G-alpha-i1. 2. The antisense compound of claim 1 which is an antisense oligonucleotide. 3. The antisense compound of claim 2 wherein the antisense oligonucleotide has a sequence comprising SEQ ID NO: 8, 11, 12, 13, 17, 18, 19, 20, 21, 22, 23, 24, 27, 29, 30, 32, 33, 34, 37, 39, 40, 41, 42, 43, 45, or 47. 4. The antisense compound of claim 2 wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage. 5. The antisense compound of claim 4 wherein the modified internucleoside linkage is a phosphorothioate linkage. 6. The antisense compound of claim 2 wherein the antisense oligonucleotide comprises at least one modified sugar moiety. 7. The antisense compound of claim 6 wherein the modified sugar moiety is a 2'-O-methoxyethyl sugar moiety. 8. The antisense compound of claim 2 wherein the antisense oligonucleotide comprises at least one modified nucleobase. 9. The antisense compound of claim 8 wherein the modified nucleobase is a 5-methylcytosine. 10. The antisense compound of claim 2 wherein the antisense oligonucleotide is a chimeric oligonucleotide. 11. A method of inhibiting the expression of human G-alpha-i1 in human cells or tissues comprising contacting said cells or tissues in vitro with the antisense compound of claim 1 so that expression of human G-alpha-i1 is inhibited. 12. An antisense compound up to 30 nucleobases in length comprising at least an 8-nucleobase portion of SEQ ID NO: 8, 12, 23, 29, 32, 33, or 39 which inhibits the expression of human G-alpha-i1. 13. The antisense compound of claim 4 which is an antisense oligonucleotide. 14. The antisense compound of claim 13 wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage. 15. The antisense compound of claim 14 wherein the modified internucleoside linkage is a phosphorothioate linkage. 16. The antisense compound of claim 13 wherein the antisense oligonucleotide comprises at least one modified sugar moiety. 17. The antisense compound of claim 16 wherein the modified sugar moiety is a 2'-O-methoxyethyl sugar moiety. 18. The antisense compound of claim 13 wherein the antisense oligonucleotide comprises at least one modified nucleobase. 19. The antisense compound of claim 18 wherein the modified nucleobase is a 5-methylcytosine. 20. The antisense compound of claim 13 wherein the antisense oligonucleotide is a chimeric oligonucleotide. 21. A method of inhibiting the expression of human G-alpha-i1 in human cells or tissues comprising contacting said cells or tissues in vitro with the antisense compound of claim 4 so that expression of human G-alpha-i1 is inhibited. 22. An antisense oligonucleotide up to 30 nucleobases in length comprisi at least an 8-nucleobase portion of SEQ ID NO: 15 which inhibits the expression of human G-alpha-i1, wherein said antisense oligonucleotide comprises phosphorothioate internucleoside linkages, 2'-methoxyethoxy wings and a deoxy gap. -------------------------------------------------------------------------------- |
| PATENT DESCRIPTION |
FIELD OF THE INVENTION The present invention provides compositions and methods for modulating the expression of G-alpha-i1. In particular, this invention relates to antisense compounds, particularly oligonucleotides, specifically hybridizable with nucleic acids encoding human G-alpha-i1. Such oligonucleotides have been shown to modulate the expression of G-alpha-i1. BACKGROUND OF THE INVENTION A vast majority of biologically active molecules including growth factors, cytokines, neurotransmitters and hormones transduce signals via specific cell-surface receptors. Some of these receptors are then coupled to heterotrimeric GTP-binding proteins (G proteins) which, upon activation, relay signals to a variety of cellular effectors including at least four phospholipase C (PLC) variants and adenylyl cyclases. G proteins mediate external signals by forming heterotrimers consisting of an alpha, beta and gamma subunit. Several isoforms of each subunit have been identified and therefore, through subunit heterogeneity, G proteins effectively integrate multiple signaling cascades. The alpha subunits of G proteins contain the GTP binding site and intrinsic catalytic GTPase activity. Based on sequence similarity and function, these subunits have been classified into four major groups; Gs, which stimulate adenylyl cyclases; Gi, which inhibit adenylyl cyclases; Gq, which activate PLC isoforms and G12/13, which mediate pathways associated with cell growth and differentiation (Hamm, J. Biol. Chem., 1998, 273, 669-672). G-alpha-i1 is a member of the Gi subfamily of G proteins which is involved in hormonal inhibition of adenylyl cyclase and in the regulation of plasma membrane enzymes. Due to the presence of all three members of this group in most cell types, the majority of studies designed to elucidate functional specificity have involved mutations, inhibitors or antibodies. Comparison studies of modified forms of the pertussis toxin-insensitive form of G-alpha-i1 (pertussis toxin is normally an inhibitor of Gi function) and the wild type protein in assays designed to investigate the interactions of adrenoceptors and Gi proteins demonstrated that more agonist was required to stimulate the mutant protein than the wild type. These studies showed that the affinity of the wild type Gi protein for the receptor was greater than that of the mutant (Wise et al., Biochem. J., 1997, 321, 721-728). Antibodies to G-alpha-i1 were used to measure the reduction of protein in brown adipose tissue when sensitized with noradrenaline. The results showed a 50% decrease in protein implicating the Gi protein in the desensitation process (Svoboda et al., Biochem. J., 1996, 314, 761-768). The expression of G-alpha-i1 has been shown to be altered in some tumors. In human hepatocellular carcinoma (HCC), the expression and functional activity of G-alpha-i1 was increased in 80% of the tumors examined. These results indicate that the regulation of the adenylate cyclase system in these cells may contribute to the formation or progression of the carcinoma (Schmidt et al., Hepatology, 1997, 26, 1189-1194). The expression of G-alpha-i1 has also been shown to be regulated by physiological shear stresses such as flow. Using a transcapillary coculture system, it was shown that G-alpha-i1 expression was decreased by high-flow conditions in endothelial and vascular smooth muscle cells. These results demonstrate that Gi proteins can mediate flow-induced responses of vessel wall dynamics which may contribute to pathological conditions such as atherosclerosis and reperfusion injury (Redmond et al., Arterioscler. Thromb. Vasc. Biol., 1998, 18, 75-83). Currently, there are no known therapeutic agents which effectively inhibit the synthesis of G-alpha-i1. Consequently there remains a long felt need for additional agents capable of effectively inhibiting G-alpha-i1 function and antisense oligonucleotides may provide a promising new pharmaceutical tool for the effective and specific modulation of G-alpha-i1 expression. SUMMARY OF THE INVENTION The present invention is directed to antisense compounds, particularly oligonucleotides, which are targeted to a nucleic acid encoding G-alpha-i1, and which modulate the expression of G-alpha-i1. Pharmaceutical and other compositions comprising the antisense compounds of the invention are also provided. Further provided are methods of modulating the expression of G-alpha-i1 in cells or tissues comprising contacting said cells or tissues with one or more of the antisense compounds or compositions of the invention. Further provided are methods of treating an animal, particularly a human, suspected of having or being prone to a disease or condition associated with expression of G-alpha-i1 by administering a therapeutically or prophylactically effective amount of one or more of the antisense compounds or compositions of the invention. |
| PATENT EXAMPLES | This data is not available for free |
| PATENT PHOTOCOPY | Available on request |
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