Claims for Patent: 5,994,315

Summary for Patent: 5,994,315

Title:	Low adenosine agent, composition, kit and method for treatment of airway disease
Abstract:	An oligonucleotide which is antisense to a mRNA encoding a polypeptide involved in airway disease(s) contains up to three adenosines per every 21 nucleotide a method of treating airway disease in a subject in need of such treatment comprises topically administering to the subject an antisense oligonucleotide in an amount effective to treat the airway disease, where the antisense oligonucleotide is essentially free of adenosine. Pharmaceutical formulations are also disclosed.
Inventor(s):	Nyce; Jonathan W. (Greenville, NC), Metzger; W. James (Greenville, NC)
Assignee:	East Carolina University (Greenville, NC)
Application Number:	08/474,497
Patent Claims:	1. A pharmaceutical composition, comprising an oligonucleotide (oligo) in aerosol form, which is effective for aleviating bronchoconstriction or lung inflammation when administered to a mammal; wherein the oligo contains no more than 3 adenosines (A) or, if at least 21 nucleotides long, it contains no more than 3 per every 21 nucleouides; and is antisense to the initiation codon, the coding region or the 5' or 3' intron-exon junctions of a gene encoding an adenosine A.sub.1 receptor, adenosine A.sub.2a receptor, adenosine A.sub.2b receptor, or adenosine A.sub.3 receptor, or anti-sense to their respective mRNAs; and a pharmaceutically acceptable carrier. 2. The composition of claim 1, wherein the oligo comprises at least one mononucleotide linking residue selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamnide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 3. The composition of claim 2, wherein all mononucleotide linking residues of the oligo are selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 4. The composition of claim 3, wherein all nucleotide linking residues are phosphorothioates. 5. The composition of claim 1, wherein the oligo is antisense to the initiation codon, the coding region or the 5' or 3' intron-exon junctions of a gene encoding an adenosine A.sub.1 receptor or anti-sense to its mRNA. 6. The composition of claim 5, wherein all nucleotide linking residues are phosphorothioates. 7. The composition of claim 1, wherein the oligo is antisense to the initiation codon, the coding region or the 5' or 3' intron-exon junctions of a gene encoding an adenosine A.sub.2a receptor or anti-sense to its mRNA. 8. The composition of claim 7, wherein all nucleotide linking residues are phosphorothioates. 9. The composition of claim 1, wherein the oligo is antisense to the initiation codon, the coding region or the 5' or 3' intron-exon junctions of a gene encoding an adenosine A.sub.2b receptor or anti-sense to its mRNA. 10. The composition of claim 9, wherein all nucleotide linking residues are phosphorothioates. 11. The composition of claim 1, wherein the oligo is antisense to the initiation codon, the coding region or the 5' or 3' intron-exon junctions of a gene encoding an adenosine A.sub.3 receptor or anti-sense to its mRNA. 12. The composition of claim 11, wherein all nucleotide linking residues are phosphorothioates. 13. The composition of claim 1, wherein the oligo is a DNA. 14. The composition of claim 1, wherein the oligo is an RNA. 15. The composition of claim 1, wherein the oligo comprises about 10 to up to about 60 mononucleotides. 16. The composition of claim 1, wherein the oligo comprises about 18 up to about 21 mononucleotides. 17. The composition of claim 1, wherein the oligo consists of SEQ ID NO.: 7; or SEQ ID NO.: 7; wherein at least one mononucleotide linking residue is selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfaimate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylarnine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 18. The composition of claim 17, wherein all nucleotide linking residues are phosphorothioates. 19. The composition of claim 1, wherein the oligo consists of SEQ ID NO.: 8; or SEQ ID NO.: 8; wherein at least one mononucleotide linking residue is selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonarmide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 20. The composition of claim 19, wherein all nucleotide linking residues are phosphorothioates. 21. The composition of claim 1, wherein the oligo is linked to a molecule selected from the group consisting of transferrin, asialoglycoprotein and streptavidin. 22. The composition of claim 1, wherein the oligo is at least 18 nucleotides long and contains no more than 1 adenosine per every 18 nucleotide. 23. The composition of claim 1, wherein the oligo is an adenosine-free oligo. 24. The composition of claim 1, further comprising an agent selected from the group consisting of antioxidants, surfactants, flavoring agents, volatile oils, buffering agents, dispersants, propellants and preservatives. 25. The composition of claim 1, wherein said pharmaceutical composition is prepared for dispensing from a pressurized aerosol dispenser. 26. The composition of claim 1, wherein said pharmaceutical composition is prepared for dispensing from a metered dose inhaler. 27. The composition of claim 1, wherein the carrier is selected from the group consisting of solid and liquid carriers. 28. The composition of claim 27, wherein the aerosol comprises a solution or suspension of the oligo. 29. The composition of claim 26, wherein said pharmaceutical composition is in the form of a spray. 30. The composition of claim 29, further comprising a buffer and a further agent selected from the group consisting of antioxidants, surfactants, flavoring agents, volatile oils, buffering agents, dispersants, propellants and preservatives. 31. The composition of claim 29, which is a respirable formulation, wherein the carrier is a respirable carrier, and which may further comprise an agent selected from the group consisting of antioxidants, flavoring agents, volatile oils, buffering agents, dispersants, surfactants, propellants and preservatives. 32. The respirable composition of claim 31, wherein at least one of the mononucleotide linking residues of the oligo is selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 33. The composition of claim 1, wherein the oligo is present in an amount of about 0.1 to about 100% w/w of the composition. 34. The composition of claim 33, wherein the oligo is present in an amount of about 0.1 to about 40% w/w of the composition. 35. The composition of claim 34, wherein the oligo is present in an amount of about 0.1 up to about 20% w/w of the composition. 36. The composition of claim 1, wherein the carrier comprises a hydrophobic carrier. 37. The composition of claim 36, wherein the carrier comprises lipid particles or vesicles. 38. The composition of claim 37, wherein the vesicles comprise liposomes and the particles comprise microcrystals. 39. The composition of claim 37, wherein the vesicles comprise liposomes which comprise the antisense oligo. 40. The composition of claim 37, wherein the particles comprise a lipid selected from the group consisting of N-(1- (2, 3-dioleoxyloxi) propyl)-N, N, N-trimethyl-ammoniummethylsulfate. 41. The composition of claim 1, comprised in a capsule or cartridge. 42. The composition of claim 1, comprising solid or liquid particles of the oligo. 43. The composition of claim 1, comprising a suspension or solution of the oligo. 44. The composition of claim 43, wherein the oligo is suspended or dissolved in a solvent or mixture of solvents. 45. The composition of claim 44, wherein the solvent is chlorofluorocarbons or chlorofluorocarbons with co-solvents, and the pharmaceutical composition further comprises an agent selected from the group consising of surfactants, antioxidants and flavoring agents. 46. The composition of claim 1, further comprising a surfactant. 47. A method of treating an adenosine receptor mediated respiratory disease or condition associated with bronchoconstriction or lung inflammation, comprising administering directly to the respiration of a mammalian subject in need of such treatment the pharmaceutical composition of claim 1, comprising an amount of the oligo effective for alleviating bronchoconstriction and/or lung inflammation. 48. The method of claim 47, wherein the pharmaceutical composition comprises respirable particles comprising the oligo. 49. The method of claim 47, wherein the disease or condition comprises lung inflammation. 50. The method of claim 47, wherein the disease or condition comprises a disease or condition asociated with bronchoconstriction. 51. The method of claim 47, wherein the disease or condition comprises asthma. 52. The method of claim 47, wherein the mammalian subject is non-human. 53. The method of clam 47, wherein the mammalian subject is a human. 54. The method of claim 47, wherein the oligo is administered in an amount of about 0.01 to about 150 mg/kg body weight. 55. The method of claim 54, wherein the oligo is administered in an amount of about 1 to about 100 mg/kg body weight. 56. The method of claim 55, wherein the oligo is administered in an amount of about 10 up to about 50 mg/kg body weight. 57. The method of claim 47, being a prophylactic method. 58. The method of claim 47, being a therapeutic method. 59. The method of claim 47, wherein the pharmaceutical composition further comprises an agent selected from the group consisting of antioxidants, flavoring agents, volatile oils, buffering agents, dispersants, surfactants, propellants and preservatives. 60. The method of claim 47, wherein the oligo is antisense to the coding region or the initiation codon of a gene encoding the adenosine receptor or antisense to an adenosine receptor mRNA; wherein at least one mononucleotide linking residue is selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 61. The method of claim 60, wherein all mononucleotide linking residues of the oligo are selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 62. The method of claim 61, wherein the pharmaceutical composition further comprises a surfactant. 63. The method of claim 47, wherein the oligo is SEQ. ID NO:7; or SEQ. ID NO:7, wherein at least one mononucleotide linking residue is selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 64. The method of claim 63, wherein the oligo is SEQ. ID NO:7, wherein all mononucleotide linking residues are selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 65. The method of claim 64, wherein all mononucleotide linking residues are phosphorothioate residues. 66. The method of claim 63, wherein the oligo is selected from SEQ. ID NO: 8; or SEQ. ID NO: 8, wherein at least one mononucleotide linking residue is selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 67. The method of claim 66, wherein all mononucleotide linking residues are selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 68. The method of claim 67, wherein all mononucleotide linking residues are phosphorothioates. 69. An in vivo method of delivering an oligonucleotide (oligo) to a target adenosine receptor polynucleotide, comprising administering into a mammalian subject's respiration an aerosol of the composition of claim 1, comprising an amount of the adenosine receptor oligo effective to reach the target adenosine receptor polynucleotide. 70. The method of claim 69, wherein the aerosol comprises respirable oligo particles. 71. The method of claim 69, wherein the oligo is oligos which are antisense to an intron-exon junction of an adenosine receptor gene selected from the group consisting of adenosine A1, A2a, A2b and A3 receptors or antisense to the corresponding adenosine receptor mRNA; or oligos which are antisense to an intron-exon junction of an adenosine receptor gene selected from the group consisting of adenosine A1, A2a, A2b and A3 receptors or antisense to the corresponding adenosine receptor mRNA, wherein the oligo comprises at least one mononucleotide linking residue selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 72. The method of claim 69, wherein the oligo is delivered to alleviate a disease or condition associated with bronchoconstriction. 73. The method of claim 72, wherein the disease or condition comprises asthma. 74. The method of claim 69, wherein the mammalian subject is a human. 75. The method of claim 69, wherein the mammalian subject is an non-human mammal. 76. The method of claim 69, wherein the oligo is administered in an amount of about 0.01 to about 150 mg/kg body weight. 77. The method of claim 76, wherein the oligo is administered in an amount of about 1 to about 100 mg/kg body weight. 78. The method of claim 77, wherein the oligo is administered in an amount of about 1 up to about 50 mg/kg body weight. 79. The method of claim 69, being a prophylactic method. 80. The method of claim 69, being a therapeutic method. 81. The method of claim 69, wherein the composition further comprises an agent selected from the group consisting of antioxidants, flavoring agents, volatile oils, buffering agents, dispersants, surfactants, propellants and preservatives. 82. The method of claim 81, wherein the composition further comprises a surfactant. 83. The method of claim 69, wherein the oligo is SEQ. ID NO: 7, and at least one mononucleotide linking residue is selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfarnate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene (methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 84. The method of claim 83, wherein all mononucleotide linking residues are selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene (methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 85. The method of claim 84, wherein all mononucleotide linking residues are phosphorothioates. 86. The method of claim 69, wherein the oligo is SEQ. ID NO: 8, wherein at least one mononucleotide linking residue is selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene (methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 87. The method of claim 86, wherein all mononucleotide linking residues are selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene (methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 88. The method of claim 87, wherein all mononucleotide linking residues are phosphorothioates. 89. The method of claim 69, wherein the oligo is oligos which are antisense to the coding region of a gene encoding an adenosine receptor selected from the group consisting of adenosine A1, A2a, A2b and A3 receptors, or antisense to the corresponding adenosine receptor mRNA; or oligos which are antisense to the coding region of a gene encoding an adenosine receptor selected from the group consisting of adenosine A1, A2a, A2b and A3 receptors, or antisense to the corresponding adenosine receptor mRNA, wherein at least one mononucleotide linking residue is selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene (methyimino), methyleneoxy (methylimino) and phosphoramidate residues. 90. A method of alleviating an airway disease or condition associated with adenosine receptor mediated bronchoconstriction or lung inflammation, comprising conducting the method of claim 89, by administering a bronchoconstriction or lung inflammation reducing amount of the oligo. 91. The method of claim 90, wherein the amount of oligo administered is effective to reduce either the level of the mRNA encoding the polypeptide or of the mRNA-encoded polypeptide, or to effect beneficial changes in the growth or characteristics of lung cells. 92. The method of claim 69, wherein the composition is dispensed from a pressurized aerosol dispenser. 93. The method of claim 69, wherein the composition is dispensed from a metered dose inhaler. 94. The method of claim 90, wherein the airway disease is associated with bronchoconstriction. 95. The method of claim 90, wherein the airway disease is associated with inflammation. 96. The method of claim 90, wherein the airway disease is associated with asthma. 97. The method of claim 90, wherein the subject is a mammal. 98. The method of claim 97, wherein the mammal is a non-human mammal. 99. The method of claim 97, wherein the mammal is a human. 100. The method of claim 90, wherein the agent is administered in an amount of about 0.01 to about 150 mg/kg body weight. 101. The method of claim 90, which is a prophylactic method. 102. The method of claim 90, which is a therapeutic method. 103. The method of claim 47, wherein the disease or condition comprises a disease or condition associated with lung inflammation. 104. The method of claim 71, wherein all the mononucleotide linking residues are selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene(methyirmino), methyleneoxy (methylimino) and phosphoramidate residues. 105. The method of claim 69, wherein the oligo is delivered to alleviate a disease or condition associated with lung inflammation.

Details for Patent 5,994,315

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2039-02-26