Oligonucleotide Core Carrier Compositions for Delivery of Nucleic Acid-Containing Therapeutic Agents, Methods of Making and Using the Same

• US 20090053169A1
• Filed: 08/19/2008
• Published: 02/26/2009
• Est. Priority Date: 08/20/2007
• Status: Active Grant

• Current Assignee: Pharmain Corporation
• Original Assignee: Pharmain Corporation
• Inventors: Reichstetter, Sandra, Bogdanov, Alexei A. JR., Bolotin, Elijah M., Castillo, Gerardo M.
• Granted Patent: US 8,563,527 B2
• Time in Patent Office: Days
• US Class Current: 424/85.200
• CPC Class Codes: A61K 31/7105 Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 31/711 Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/6455 Polycationic oligopeptides, polypeptides or polyamino acids, e.g. for complexing nucleic acids
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

The present invention relates, in part, to an oligonucleotide-core carrier comprising a carrier, and oligonucleotide groups covalently linked to the carrier. The oligonucleotide groups are capable of dissociably linking load molecules such as therapeutic agents. The oligonucleotide-core carrier may also comprise protective side chains, and targeting molecules.

Claims

1. A oligonucleotide-core carrier composition comprising (i) a carrier;

(ii) an oligonucleotide group capable of binding a load molecule, wherein the oligonucleotide group is covalently linked to the carrier, and each has less than 100 bases; and

(iii) a plurality of polymeric protective side chains, wherein each protective side chain is covalently linked to the carrier or the oligonucleotide of the carrier and each protective side chains has a molecular weight between about 400 and 20,000 Daltons independent of the carrier and oligonucleotide weight.

2. The oligonucleotide-core carrier composition of claim 1, wherein the oligonucleotide comprises various nucleotide monomers selected from a group consisting of 2′

-deoxyribonucleotide, ribonucleotide, 2′

-O-methylribonucleotide, locked ribonucleotide, N-(2-ethylamino)glycine nucleotide and morpholino nudeotide, wherein each nucleotide is covalently linked to another by 3′

-5′

or 2′

-5′

linkage, wherein the linkage comprises of a phosphodiester, phosphorothio, phosphotriester, phosphorodiamidate and a peptide, wherein the base component of nucleotide is anyone of adenine, thymidine, guanine, cytosine, uracil, inosine, thioinosine, thiouridine, xanthosine, pseudouridine, or orotidine.

3. The oligonucleotide-core carrier composition of claim 1, wherein the protective side chain is polyethylene glycol, polypropylene glycol, a co-polymer of polyethylene glycol and polypropylene glycol, polysaccharide, or alkoxy derivatives thereof.

4. The oligonucleotide-core carrier composition of claim 3, wherein the alkoxy derivative is methoxypolyethylene glycol, methoxypolypropylene glycol, a methoxylated co-polymer of polyethylene glycol and polypropyleneglycol, or ethoxylated polysaccharide.

5. The oligonucleotide-core carrier composition of claim 1, wherein the protective side chain is methoxypolyethylene glycol.

6. The oligonucleotide-core carrier composition of claim 1, further comprising a load molecule, wherein the load molecule has oligonucleotide dissodably linked by hydrogen bonding to the oligonucleotide group of the carrier.

7. The oligonucleotide-core carrier composition of claim 6, wherein the load molecule is a therapeutic agent.

8. The oligonucleotide-core carrier composition of claim 7, wherein the therapeutic agent is selected from the group consisting of siRNA, miRNA, antisense DNA, antisense RNA, oligonucleotide aptamers, and oligonucleotide-bonded peptide/proteins;

wherein the peptide/protein is selected from a group consisting of peptide aptamer, glucagon-like-peptide, glucagon-like-peptide derivative, exenatide, leptin, leptin fragment, alpha-melanocyte stimulating hormone, adeponectin, Gastric inhibitory polypeptide(GIP), Epidermal Growth Factor (EGF) receptor ligand, EGF, Tranforming Growth Factor alpha (TGF-alpha), Betacellulin, Gastrin/Cholecystokinin receptor ligand, Gastrin, Cholecystokinin, lysostaphin, interferon, interferon gamma, interferon beta, interferon alpha, interleukin-1, interleukin-2, interleukin-4, interleukin-6, interleukin-8, interleukin-10, interleukin-12, tumor necrosis factor, tumor necrosis factor alpha, tumor necrosis factor beta, insulin, insulin-like growth factor, growth hormone, nerve growth factor, brain-derived neurotrophic factor, enzymes, endostatin, angiostatin, trombospondin, urokinase, streptokinase, blood clotting factor VII, blood clotting factor VIII, granulucyte-macrophage colony-stimulating factor (GM-CSF), granulucyte colony-stimulating factor (G-CSF), thrombopoetin, calcitonin, parathyroid hormone (PTH) and its fragments, erythropoietin, atrial natriuretic factor, monoclonal antibodies, monoclonal antibody fragments, somatostatin, protease inhibitors, adrenocorticotropin, gonadotropin releasing hormone, oxytocin, leutinizing-hormone-releasing-hormone, follicle stimulating hormone, glucocerebrosidase, thrombopoietin, and filgrastin.

9. The oligonucleotide-core carrier composition of claim 7, wherein the therapeutic agent is siRNA, miRNA, antisense DNA, antisense RNA, oligonucleotide aptamers, and oligonucleotide-bonded peptide/proteins;

wherein the peptide/protein is selected from a group consisting of Glucagon-like-peptide, Leptin, Leptin fragment, Peptide YY, Epidermal Growth Factor (EGF) receptor ligand, EGF, Tranforming Growth Factor alpha (TGF-alpha), Betacellulin, Gastrin/Cholecystokinin receptor ligand, Gastrin, Cholecystokinin, interferon, factor VIII, terlipressin, Gastric inhibitory polypeptide(GIP), and vasoactive intestinal peptide (VIP).

10. The oligonucleotide carrier composition of claim 7, wherein the therapeutic agent is siRNA against anyone from the group consisting of Ghrelin, NPY, Cathepsin, Myostatin, TSLP, IL-4, IL-8, L-12, IL-13, STAT-6, MIP-1 alpha, RANTES, CCR1, CCR3, INF-gamma, TNF-alpha, ICXCR1, MCP-1, and CCR2, CXCR3, CXCR4, CXCR5, CCR4, CCR5, CCR6, CCR7, CCR8, gp120, gp41, p17, p24, RT, HIV proteases, Fas (CD95), FAS-L, FADD, Caspase-8, IL-1, IL-6, Bak, Bax, Bid, Bcl-2, Bcl-XL, HLA-G, IGF-1, EGF, FGF, VEGF, VEGFR, IGFR, EGFR, FGFR, HER2, TGF-beta, Caspase 3, CEACAM6, HPV-E6, HPV-E7, H-Ras gene, P100a gene, CREB, BRAF gene, ATF2, N-myc gene, Cox1, Cox2, GluR2, DAT, VEGFR1, TGF-b-RII, IL-1-beta, Facipain-1-2 malaria protein, viral Capsid protein, NS5A, NP influenza protein, PA influenza protein, and HBV.

11. The oligonucleotide-core carrier composition of claim 1, further comprising a targeting molecule covalently linked to the protective side chains.

12. The oligonucleotide-core carrier composition of claim 11, wherein the targeting molecule is selected from a group consisting of an antibody, fragment of an antibody, chimeric antibody, lectins, receptor ligands, proteins, enzymes, peptides, saccharides, quasi substrates of enzymes, cell-surface-binding compounds, and extracellular-matrix-binding compounds.

13. The biocompatible oligonucleotide-core carrier composition of claim 11, further comprising a load molecule, wherein the load molecule has oligonucleotide dissociably linked by hydrogen bonding to the oligonucleotide group of the carrier.

14. An oligonucleotide-core carrier composition, comprising:

(i) a carrier; and

(ii) an oligonucleotide group with a covalently linked protective side chain, wherein the oligonucleotide group has a first and second end;

the first end is covalently linked to the carrier, the second end is covalently linked to the said protective side chain;

the oligonucleotide group has 3-100 bases, and;

the protective side chain has a molecular weight between about 460 and 20,000 Daltons independent of the carrier and oligonucleotide group weights.

15. The oligonucleotide-core carrier composition of claim 14, wherein the protective side chain is polyethylene glycol, polypropylene glycol, a co-polymer of polyethylene glycol and polypropylene glycol, polysaccharide, or alkoxy derivatives thereof.

16. The oligonucleotide-core carrier composition of claim 15, wherein the alkoxy derivative is methoxypolyethylene glycol, methoxypolypropylene glycol, a methoxylated co-polymer polyethylene glycol and polypropyleneglycol, or ethoxylated polysaccharide.

17. The oligonucleotide-core carrier composition of claim 16, wherein the alkoxy derivative is is methoxypolyethylene glycol.

18. The oligonucleotide-core carrier composition of claim 14, further comprising a load molecule, wherein the load molecule has oligonucleotide dissociably linked by hydrogen bonding to the oligonucleotide group of the carrier.

19. The oligonucleotide carrier composition of claim 18, wherein the load molecule is a therapeutic agent.

20. The oligonucleotide carrier composition of claim 19, wherein the therapeutic agent is selected from the group consisting of siRNA, miRNA, antisense DNA, antisense RNA, oligonucleotide aptamers, and oligonucleotide-bonded peptide/proteins;

wherein the peptide/protein is selected from a group consisting of peptide aptamer, glucagon-like-peptide, glucagon-like-peptide derivative, exenatide, leptin, leptin fragment, alpha-melanocyte stimulating hormone, adeponectin, Gastric inhibitory polypeptide(GIP), Epidermal Growth Factor (EGF) receptor ligand, EGF, Tranforming Growth Factor alpha (TGF-alpha), Betacellulin, Gastrin/Cholecystokinin receptor ligand, Gastrin, Cholecystokinin, lysostaphin, interferon, interferon gamma, interferon beta, interferon alpha, interleukin-1, interleukin-2, interleukin-4, interleukin-6, interleukin-8, interieukin-10, interleukin-12, tumor necrosis factor, tumor necrosis factor alpha, tumor necrosis factor beta, insulin, insulin-like growth factor, growth hormone, nerve growth factor, brain-derived neurotrophic factor, enzymes, endostatin, angiostatin, trombospondin, urokinase, streptokinase, blood clotting factor VII, blood clotting factor VIII, granulucyte-macrophage colony-stimulating factor (GM-CSF), granulucyte colony-stimulating factor (G-CSF), thrombopoetin, calcitonin, parathyroid hormone (PTH) and its fragments, erythropoietin, atrial natriuretic factor, monoclonal antibodies, monoclonal antibody fragments, somatostatin, protease inhibitors, adrenocorticotropin, gonadotropin releasing hormone, oxytocin, leutinizing-hormone-releasing-hormone, follicle stimulating hormone, glucocerebrosidase, thrombopoietin, and filgrastin.

21. The oligonucleotide carrier composition of claim 19, wherein the therapeutic agent is siRNA against anyone from the group consisting of Ghrelin, NPY, Cathepsin, Myostatin, TSLP, IL-4, IL-8, L-12, IL-13, STAT-6, MIP-1 alpha, RANTES, CCR1, CCR3, INF-gamma, TNF-alpha, ICXCR1, MCP-1, and CCR2, CXCR3, CXCR4, CXCR5, CCR4, CCR5, CCR6, CCR7, CCR8, gp120, gp41, p17, p24, RT, HIV proteases, Fas (CD95), FAS-L, FADD, Caspase-8, IL-1, IL-6, Bak, Bax, Bid, Bcl-2, Bcl-XL, HLA-G, IGF-1, EGF, FGF, VEGF, VEGFR, IGFR, EGFR, FGFR, HER2, TGF-beta, Caspase 3, CEACAM6, HPV-E6, HPV-E7, H-Ras gene, P100a gene, CREB, BRAF gene, ATF2, N-myc gene, Cox1, Cox2, GluR2, DAT, VEGFR1, TGF-b-RII, IL-1-beta, Facipain-1-2 malaria protein, viral Capsid protein, NS5A, NP influenza protein, PA influenza protein, and HBV.

22. The oligonucleotide-core carrier composition of claim 14, further comprising a targeting molecule covalently linked to the protective side chains.

23. The oligonucleotide-core carrier composition of claim 22, wherein the targeting molecule is selected from the group consisting of an antibody, fragment of an antibody, chimeric antibody, enzymes, peptides, quasi substrates of enzymes, lectins, receptor, and saccharide ligands of lectins.

24. The oligonucleotide-core carrier composition of claim 22, further comprising a load molecule, wherein the load molecule has oligonucleotide dissodably linked by hydrogen bonding to the oligonucleotide group of the carrier.

25. The oligonucleotide-core carrier composition of claim 14, further comprising of a second oligonucleotide group covalently linked to the carrier but not to protective chains, and the said oligonucleotide has 3 to 100 bases.

26. The oligonucleotide-core carrier composition of claim 25, further comprising a load molecule, wherein the load molecule has oligonucleotide dissodably linked by hydrogen bonding to the oligonucleotide group of the carrier.

27. The oligonucleotide-core carrier composition of claim 25, further comprising a targeting molecule covalently linked to the protective side chain.

28. The oligonucleotide-core carrier composition of claim 27, further comprising a load molecule, wherein the load molecule has oligonucleotide dissociably linked by hydrogen bonding to the oligonucleotide group of the carrier.

29. A biocompatible oligonucleotide-core carrier composition comprising:

(i) a carrier wherein the carrier is a polymeric carrier selected from a group consisting of polylysine, polyaspartic acid, polyglutamic acid, polyserine, polythreonine, polycysteine, polyglycerol, polyethyleneimine, polyallyamine, chitosan, natural polysaccharide, aminated polysaccharides, aminated oligosaccharides, polyamidoamine, polyacrylic acids, polyalcohols, sulfonated polysaccharides, sulfonated oligosaccharides, carboxylated polysaccharides, carboxylated oligosaccharides, aminocarboxylated polysaccharides, aminocarboxylated oligosaccharides, carboxymethylated polysaccharides, and carboxymethylated oligosaccharides; and

(ii) an oligonucleotide group with 3 to 100 bases covalently linked to the carrier.

30. The oligonucleotide-core carrier composition of claim 29, further comprising a load molecule, wherein the load molecule has oligonucleotide dissodably linked by hydrogen bonding to the oligonucleotide group of the carrier.

31. The oligonucleotide carrier composition of claim 30, wherein the load molecule is a therapeutic agent.

32. The oligonucleotide-core carrier composition of claim 29, further comprising a targeting molecule covalently linked to the carrier or oligonucleotide groups.

33. The oligonucleotide-core carrier composition of claim 32, wherein the targeting molecule is selected from the group consisting of an antibody, fragment of an antibody, chimeric antibody, enzymes, peptides, receptor, quasi substrates of enzymes, lectins, and saccharide ligands of lectins.

34. The oligonucleotide-core carrier composition of claim 32, further comprising a load molecule, wherein the load molecule has oligonucleotide dissociably linked by hydrogen bonding to the oligonucleotide group of the carrier.

35. The oligonucleotide carrier composition of claim 34, wherein the load molecule is a therapeutic agent.

36. A method of increasing the blood circulation half-life of peripherally administered oligonucleotides by injecting to a patient a formulation comprising anyone of the compositions in above claims.