| Peptide conjugated, inosine-substituted antisense oligomer compound and method -> Monitor Keywords |
|
|
 
Peptide conjugated, inosine-substituted antisense oligomer compound and methodRelated Patent Categories: Drug, Bio-affecting And Body Treating Compositions, Designated Organic Active Ingredient Containing (doai), O-glycoside, , Nitrogen Containing Hetero Ring, Polynucleotide (e.g., Rna, Dna, Etc.)Peptide conjugated, inosine-substituted antisense oligomer compound and method description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20050288246, Peptide conjugated, inosine-substituted antisense oligomer compound and method. Brief Patent Description - Full Patent Description - Patent Application Claims
[0001] This patent application claims priority to U.S. Provisional Application No. 60/574,048 filed on May 24, 2004, which is incorporated in its entirety herein by reference. FIELD OF THE INVENTION [0002] This invention relates to an antisense oligomer compound (i) conjugated to an arginine rich-peptide effective to enhance the uptake of the oligomer into cells, and (ii) in which strings of G bases are broken by one of more inosine bases, and methods of using such compound. REFERENCES [0003] Devi, G. R. (2002). "Prostate cancer: status of current treatments and emerging antisense-based therapies." Curr Opin Mol Ther 4(2): 138-48. Hudziak, R. M., E. Barofsky, et al. (1996). "Resistance of morpholino phosphorodiamidate oligomers to enzymatic degradation." Antisense Nucleic Acid Drug Dev 6(4): 267-72. [0004] Iversen, P. L. (2001). Phosphoramidite Morpholino Oligomers. Antisense Drug Technology. S. T. Crooke. New York, Marcel Dekker, Inc. Shafer, R. H. and I. Smirnov (2000). "Biological aspects of DNA/RNA quadruplexes." Biopolymers 56(3): 209-27. [0005] Stein, D. A., D. E. Skilling, et al. (2001). "Inhibition of Vesivirus infections in mammalian tissue culture with antisense morpholino oligomers." Antisense Nucleic Acid Drug Dev 11(5): 317-25. [0006] Summerton, J. and D. Weller (1997). "Morpholino antisense oligomers: design, preparation, and properties." Antisense Nucleic Acid Drug Dev 7(3): 187-95. [0007] Vanin, E. F. and T. H. Ji (1981). "Synthesis and application of cleavable photoactivable heterobifunctional reagents." Biochemistry 20(24): 6754-60. [0008] Dapic, V., V. Abdomerovic, et al. (2003). "Biophysical and biological properties of quadruplex oligodeoxyribonucleotides." Nucleic Acids Res 31(8): 2097-107. [0009] Kang, S. H., M. J. Cho, et al. (1998). "Up-regulation of luciferase gene expression with antisense oligonucleotides: implications and applications in functional assay development." Biochemistry 37(18): 6235-9. [0010] Knapp, D. C., J. E. Mata, et al. (2003). "Resistance to chemotherapeutic drugs overcome by c-Myc inhibition in a Lewis lung carcinoma murine model." Anticancer Drugs 14(1): 39-47. [0011] Shafer, R. H. and I. Smirnov (2000). "Biological aspects of DNA/RNA quadruplexes." Biopolymers 56(3): 209-27. [0012] Vanin, E. F. and T. H. Ji (1981). "Synthesis and application of cleavable photoactivable heterobifunctional reagents." Biochemistry 20(24): 6754-60. BACKGROUND OF THE INVENTION [0013] Antisense oligomers offer great potential as pharmaceutical drugs, as evidenced by the number of antisense drugs currently in clinical development, and aided by the fact that a number of potential limitations of antisense oligomers have been successfully addressed over the past several years (Devi, Stein). Novel uncharged oligomer backbones have been developed to improve uptake into cells, and to increase resistance to nuclease degradation (Hudziak, Iversen, Summerton). For some oligomer structures, for example, morpholino based structures, the modified backbone has been found to give enhanced binding affinity to its target nucleic acid (Iversen, Summerton). [0014] More recently, it has been discovered that a variety of arginine-rich peptides can dramatically increase the level of uptake of uncharged oligonucleotides into cells, including mammalian cells (see, for example, co-owned U.S. patent application Ser. No. 60/466,703, filed Apr. 29, 2003, and corresponding U.S. patent application for "Compositions for Enhancing Transport of Molecules into Cells," filed Apr. 29, 2004, both of which are incorporated herein in its entirety). This discovery has the potential to significantly increase the therapeutic potential of a variety of antisense oligomers, including those intended to block expression of selected proteins, those aimed at blocking certain donor or acceptor splice sites in pre-processed mRNA, and those designed to treat viral infection by blocking expression of viral genes or replication of single-stranded viral genomes. [0015] In some antisense applications, the optimal targeting sequence against which the oligomer antisense is directed may include a run of four of more cytosine bases, in which case the oligomer will contain a corresponding string of four or more complementary guanine bases. As an important example, an optimal target sequence for the c-myc protein is a region containing the AUG start site of the c-myc RNA that includes a run of four cytosine bases. Anti-sense oligomers directed against the start-codon region of c-myc have a number of important therapeutic applications, including the treatment of cancer, polycistic kidney disease (see, for example, co-owned U.S. Pat. No. 6,875,747, which is incorporated herein in its entirety), coronary-vessel restenosis (see, for example, co-owned PCT patent application WO00/44897, published Aug. 3, 2000, which is incorporated herein in its entirety), and cancer therapy (see, for example, co-owned U.S. patent application US-2003-0087861-A1, published May 8, 2003, which is incorporated herein by reference in its entirety.) [0016] Surprisingly, it has now been found that conjugating an arginine-rich peptide to antisense compounds having runs of four or more cytosine bases, in an effort to enhance the cellular uptake of the oligomer, severely compromises the antisense activity of the compound, as well as the ability to purify the compound. Although the basis of this problem is not understood, it appears to involve an interaction between the positively charged peptide and the oligomer compound in a way that promotes the formation of G-quartets in the oligomer, thus reducing the solubility and/or ability of the compound to bind to its target nucleic acid. It would therefore be useful to enhance the cellular uptake of such antisense oligomer compounds, by conjugating the compound with an arginine-rich peptide, without degrading the antisense activity of the compound with respect to its intracellular target. [0017] In particular, it would be useful to enhance the cellular uptake of the above c-myc antisense compound without loss of antisense activity, for purposes of enhancing the compound's therapeutic activity in the treatment of cancer, polycistic kidney disease or coronary-vessel restenosis. SUMMARY OF THE INVENTION [0018] The method includes, in one aspect, an improvement in a method for enhancing the cellular uptake of a substantially uncharged oligonucleotide analog compound, by forming a conjugate of the compound and an arginine-rich peptide effective to enhance the uptake of the compound into target cells, where the compound includes a string of bases that are complementary to four or more contiguous cytosine bases in a target nucleic acid region to which the compound is intended to bind. The improvement includes substituting an inosine base for at least one guanine base in the string of bases in the compound so as to limit the number of contiguous guanine bases in the string to three or fewer, preferably two or fewer. [0019] The improvement may be effective to enhance the water solubility of the conjugate during a purification step involving conjugate binding to and release from an cationic ion exchange resin, relative to the same conjugate in the absence of the inosine substitution. Where the target nucleic acid region includes the start codon in an mRNA, the improvement may be effective to enhance the ability of the conjugate to block translation of the protein encoded by the mRNA, relative to the same conjugate in the absence of the inosine substitution. Where the target nucleic acid region includes a donor or acceptor splice site in an preprocessed mRNA, the improvement may be effective to enhance the ability of the conjugate to mask mRNA splicing at the target region, relative to the same conjugate in the absence of the inosine substitution. [0020] Where the target nucleic acid region includes a virally-encoded cis-acting element involved in viral replication, the improvement may be effective to enhance the ability of the conjugate to block viral replication, relative to the same conjugate in the absence of the inosine substitution. Continue reading about Peptide conjugated, inosine-substituted antisense oligomer compound and method... Full patent description for Peptide conjugated, inosine-substituted antisense oligomer compound and method Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Peptide conjugated, inosine-substituted antisense oligomer compound and method patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. Each week you receive an email with patent applications related to your keywords. Start now! - Receive info on patent apps like Peptide conjugated, inosine-substituted antisense oligomer compound and method or other areas of interest. ### Previous Patent Application: Oligonucleotides comprising a c5-modified pyrimidine Next Patent Application: Peptides acting as both glp-1 receptor agonists and glucagon receptor antagonists and their pharmacological methods of use Industry Class: Drug, bio-affecting and body treating compositions ### FreshPatents.com Support Thank you for viewing the Peptide conjugated, inosine-substituted antisense oligomer compound and method patent info. IP-related news and info Results in 0.13133 seconds Other interesting Feshpatents.com categories: Software: Finance , AI , Databases , Development , Document , Navigation , Error 174 |
 * Protect your Inventions * US Patent Office filing 
PATENT INFO |
|
