Formulation of adenovirus for gene therapy

Abstract

The present invention addresses the need to improve the long-term storage stability (i.e. infectivity) of vector formulations. In particular, it has been demonstrated that for adenovirus, the use of bulking agents, cryoprotectants and lyoprotectants imparts desired properties that allow both lyophilized and liquid adenovirus formulations to be stored at 4° C. for up to 6 months and retain an infectivity between 60-100% of the starting infectivity.

1 - 60 . (canceled)  
     
     
         61 . A pharmaceutical composition in an oral dosage form, wherein the oral dosage form is a tablet, a troche, a capsule, an elixir, a suspension, a syrup, a solution, a wafer, a pill, a microsphere, a nanoparticle, a mouthwash, a dentrifice, or a spray, wherein the composition comprises a viral vector comprising an expression cassette, wherein the expression cassette comprises a promoter active in eukaryotic cells, wherein the promoter is operatively coupled to a nucleic acid encoding a therapeutic gene.  
     
     
         62 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a tablet.  
     
     
         63 . The pharmaceutical composition of  claim 62 , wherein the tablet is an ingestible tablet or a buccal tablet.  
     
     
         64 . The pharmaceutical composition of  claim 62 , wherein the tablet is formulated to dissolve in the mouth or under the tongue.  
     
     
         65 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a troche.  
     
     
         66 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a capsule.  
     
     
         67 . The pharmaceutical composition of  claim 66 , wherein the capsule is a hard-shell capsule or a soft-shell capsule.  
     
     
         68 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is an elixir.  
     
     
         69 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a suspension.  
     
     
         70 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a syrup.  
     
     
         71 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a solution.  
     
     
         72 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a wafer.  
     
     
         73 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a pill.  
     
     
         74 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a microsphere.  
     
     
         75 . The pharmaceutical composition of  claim 74 , wherein the microsphere comprises a polymer that can adhere to a mucosal surface.  
     
     
         76 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a nanoparticle.  
     
     
         77 . The pharmaceutical composition of  claim 76 , wherein the nanoparticle is formulated with a stabilizer and/or a linear poly(ethylene oxide) polymer.  
     
     
         78 . The pharmaceutical composition of  claim 76 , wherein the nanoparticle is formulated for controlled release.  
     
     
         79 . The pharmaceutical composition of  claim 76 , wherein the nanoparticles comprise a biodegradable polycyanoacrylate polymer.  
     
     
         80 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a mouthwash.  
     
     
         81 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a dentrifice.  
     
     
         82 . The pharmaceutical composition of  claim 81 , wherein the dentrifice is a gel, a paste, a powder, or a slurry.  
     
     
         83 . The pharmaceutical composition of  claim 82 , further comprising water, a binder, an abrasive, a flavoring agent, a foaming agent, and/or a humectant.  
     
     
         84 . The pharmaceutical composition of  claim 61 , wherein the oral dosage form is a spray.  
     
     
         85 . The pharmaceutical composition of  claim 61 , further comprising an inert diluent, an assimilable edible carrier, a binder, an excipient, a disintegrating agent, a lubricant, a sweetening agent, a flavoring agent, a liquid carrier, a coating, a preservative, a dye, a solvent, water, an abrasive, a foaming agent, a humectant, and/or an adhesive.  
     
     
         86 . The pharmaceutical composition of  claim 85 , wherein the binder is gum tragacanth, acacia, constarch, or gelatin.  
     
     
         87 . The pharmaceutical composition of  claim 85 , wherein the excipient is dicalcium phosphate, glycerin, or potassium bicarbonate.  
     
     
         88 . The pharmaceutical composition of  claim 85 , wherein the disintegrating agent is corn starch, potato starch, or alginic acid.  
     
     
         89 . The pharmaceutical composition of  claim 85 , wherein the lubricant is magnesium stearate.  
     
     
         90 . The pharmaceutical composition of  claim 85 , wherein the sweetening agent is sucrose, lactose, or saccharin.  
     
     
         91 . The pharmaceutical composition of  claim 85 , wherein the flavoring agent is peppermint, oil of wintergreen, or cherry flavoring.  
     
     
         92 . The pharmaceutical composition of  claim 85 , wherein the oral dosage form is a tablet, pill, or capsule, and wherein the coating is shellac or sugar.  
     
     
         93 . The pharmaceutical composition of  claim 61 , wherein the adenovirus vector is formulated to be incorporated directly into food.  
     
     
         94 . The pharmaceutical composition of  claim 61 , further defined as a sustained release preparation or controlled release preparation.  
     
     
         95 . The pharmaceutical composition of  claim 85 , wherein the solvent comprises sodium borate solution.  
     
     
         96 . The pharmaceutical composition of  claim 61 , wherein the viral vector is an adenoviral vector, a retroviral vector, an adeno-associated virus vector, a vaccinia virus vector, a herpesvirus vector, or an oncolytic virus vector.  
     
     
         97 . The pharmaceutical composition of  claim 96 , wherein the viral vector is an adenoviral vector.  
     
     
         98 . The pharmaceutical composition of  claim 97 , wherein the adenoviral vector is a replication-deficient adenoviral vector.  
     
     
         99 . The pharmaceutical composition of  claim 98 , wherein the replication-deficient adenoviral vector is lacking at least a portion of the E1 region.  
     
     
         100 . The pharmaceutical composition of  claim 96 , wherein the oncolytic virus is a reovirus, OYNX-015, or CN706.  
     
     
         101 . The pharmaceutical composition of  claim 61 , wherein the promoter is a CMV IE, SV40 early, or RSV LTR.  
     
     
         102 . The pharmaceutical composition of  claim 61 , wherein the viral vector is comprised within a liposome.  
     
     
         103 . The pharmaceutical composition of  claim 61 , wherein the therapeutic gene is a tumor suppressor gene, a gene encoding an inducer of apoptosis, a gene encoding an enzyme, a gene encoding a hormone, a gene encoding an interleukin, or a gene encoding a cytokine.  
     
     
         104 . The pharmaceutical composition of  claim 103 , wherein the tumor suppressor gene is p53, CDK4 or other cyclin-dependent kinase; p16 INK4 , p16 B ,  p21WAF1, CIP1, SDI1 , p27 KIP1  or other CDK-inhibitory protein; C-CAM, RB, APC, DCC, NF-1, NF-2, WT-1, MEN-I, MEN-II, zac1, p73, BRCA1, VHL, FCC, MMAC1, MCC, p16, p21, p57, p27, and BRCA2.  
     
     
         105 . The pharmaceutical composition of  claim 103 , wherein the inducer of apoptosis is Bax, Bak, Bcl-X s , Bik, Bid, Harakiri, Ad E1B, Bad, or an ICE-CED2 protease.  
     
     
         106 . The pharmaceutical composition of  claim 103 , wherein the enzyme is cytosine deaminase, hypoxanthine-guanine phosphoribosyltransferase, galactose-1-phosphate uridyltransferase, phenylalanine hydroxylase, glucocerebrosidase, sphingomyelinase, α-L-iduronidase, glucose-6-phosphate dehydrogenase, HSV thymidine kinase, or human thymidine kinase.  
     
     
         107 . The pharmaceutical composition of  claim 103 , wherein the hormone is growth hormone, prolactin, placental lactogen, luteinizing hormone, follicle-stimulating hormone, chorionic gonadotropin, thyroid-stimulating hormone, leptin, adrenocorticotropin (ACTH), angiotensin I, angiotensin II, β-endorphin, β-melanocyte stimulating hormone, cholecystokinin, endothelin I, galanin, gastric inhibitory peptide, glucagon, insulin, a lipotropin, a neurophysin, somatostatin, calcitonin, calcitonin gene related peptide, β-calcitonin gene related peptide, hypercalcemia of malignancy factor, parathyroid hormone-related protein, glucagon-like peptide, pancreastatin, pancreatic peptide, peptide YY, PHM, secretin, vasotocin, enkephalinamide, metorphinamide, alpha melanocyte stimulating hormone, atrial natriuretic factor, amylin, amyoid P component (SAP-1), corticotropin releaseing hormone (CRH), growth hormone releasing factor (GHRH), luteinizing hormone releasing hormone (LHRH), neuropeptide Y, substance K (neurokinin A), substance P, or thryrotropin releasing hormone (TRH).  
     
     
         108 . The pharmaceutical composition of  claim 103 , wherein the interleukin or cytokine is IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, GM-CSF, or G-CSF.  
     
     
         109 . A pharmaceutical composition in an nasal dosage form comprising a viral vector comprising an expression cassette, wherein the expression cassette comprises a promoter active in eukaryotic cells, wherein the promoter is operatively coupled to a nucleic acid encoding a therapeutic gene.  
     
     
         110 . The pharmaceutical composition of  claim 109 , wherein the nasal dosage form is an intranasal aerosol spray.  
     
     
         111 . The pharmaceutical composition of  claim 110 , wherein the composition is further defined as being comprised in a liposome.  
     
     
         112 . The pharmaceutical composition of  claim 110 , wherein the intranasal aerosol spray comprises microspheres.  
     
     
         113 . The pharmaceutical composition of  claim 112 , wherein the microspheres are bioadhesive microspheres.  
     
     
         114 . The pharmaceutical composition of  claim 113 , wherein the microspheres comprise starch, gelatin, dextran, collagen, an adsorption enhancer, and/or albumin.  
     
     
         115 . The pharmaceutical composition of  claim 114 , wherein the absorption enhancer is a surfactant.  
     
     
         116 . The pharmaceutical composition of  claim 109 , wherein the nasal dosage form is an aerosol.  
     
     
         117 . The pharmaceutical composition of  claim 109 , wherein the nasal dosage form is a microparticle resin.  
     
     
         118 . The pharmaceutical composition of  claim 117 , wherein the microparticle resin is fractionated sodium polystyrene sulfonate powder or styrene-divinylbenzene copolymer.  
     
     
         119 . The pharmaceutical composition of  claim 109 , wherein the composition is formulated to be delivered via transmucosal delivery.  
     
     
         120 . The pharmaceutical composition of  claim 119 , further comprising an absorption enhancer.  
     
     
         121 . The pharmaceutical composition of  claim 120 , wherein the absorption enhancer is a lysophosphatidyl-glycerol compound.  
     
     
         122 . The pharmaceutical composition of  claim 119 , wherein the composition further comprises a polytetrafluoroethylene support matrix.  
     
     
         123 . The pharmaceutical composition of  claim 109 , wherein the viral vector is an adenoviral vector, a retroviral vector, an adeno-associated virus vector, a vaccinia virus vector, a herpesvirus vector, or an oncolytic virus vector.  
     
     
         124 . The pharmaceutical composition of  claim 123 , wherein the viral vector is an adenoviral vector.  
     
     
         125 . The pharmaceutical composition of  claim 124 , wherein the adenoviral vector is a replication-deficient adenoviral vector.  
     
     
         126 . The pharmaceutical composition of  claim 125 , wherein the replication-deficient adenoviral vector is lacking at least a portion of the E1 region.  
     
     
         127 . The pharmaceutical composition of  claim 123 , wherein the oncolytic virus is a reovirus, OYNX-015, or CN706.  
     
     
         128 . The pharmaceutical composition of  claim 109 , wherein the promoter is a CMV IE, SV40 early, or RSV LTR.  
     
     
         129 . The pharmaceutical composition of  claim 109 , wherein the viral vector is comprised within a liposome.  
     
     
         130 . The pharmaceutical composition of  claim 109 , wherein the therapeutic gene is a tumor suppressor gene, a gene that encodes an inducer of apoptosis, a gene encoding an enzyme, a gene encoding a hormone, a gene encoding an interleukin, or a gene encoding a cytokine.  
     
     
         131 . The pharmaceutical composition of  claim 130 , wherein the tumor suppressor gene is p53, CDK4 or other cyclin-dependent kinase; p16 INK4 , p16 B ,  p21WAF1, CIP1, SDI1 , p27 KIP1  or other CDK-inhibitory protein; C-CAM, RB, APC, DCC, NF-1, NF-2, WT-1, MEN-I, MEN-II, zacl, p73, BRCA1, VHL, FCC, MMAC1, MCC, p16, p21, p57, p27, and BRCA2.  
     
     
         132 . The pharmaceutical composition of  claim 130 , wherein the inducer of apoptosis is Bax, Bak, Bcl-X s , Bik, Bid, Harakiri, Ad E1B, Bad, or an ICE-CED2 protease.  
     
     
         133 . The pharmaceutical composition of  claim 130 , wherein the enzyme is cytosine deaminase, hypoxanthine-guanine phosphoribosyltransferase, galactose-1-phosphate uridyltransferase, phenylalanine hydroxylase, glucocerebrosidase, sphingomyelinase, α-L-iduronidase, glucose-6-phosphate dehydrogenase, HSV thymidine kinase, or human thymidine kinase.  
     
     
         134 . The pharmaceutical composition of  claim 130 , wherein the hormone is growth hormone, prolactin, placental lactogen, luteinizing hormone, follicle-stimulating hormone, chorionic gonadotropin, thyroid-stimulating hormone, leptin, adrenocorticotropin (ACTH), angiotensin I, angiotensin II, β-endorphin, β-melanocyte stimulating hormone, cholecystokinin, endothelin I, galanin, gastric inhibitory peptide, glucagon, insulin, a lipotropin, a neurophysin, somatostatin, calcitonin, calcitonin gene related peptide, β-calcitonin gene related peptide, hypercalcemia of malignancy factor, parathyroid hormone-related protein, glucagon-like peptide, pancreastatin, pancreatic peptide, peptide YY, PHM, secretin, vasotocin, enkephalinamide, metorphinamide, alpha melanocyte stimulating hormone, atrial natriuretic factor, amylin, amyoid P component (SAP-1), corticotropin releaseing hormone (CRH), growth hormone releasing factor (GHRH), luteinizing hormone releasing hormone (LHRH), neuropeptide Y, substance K (neurokinin A), substance P, or thryrotropin releasing hormone (TRH).  
     
     
         135 . The pharmaceutical composition of  claim 130 , wherein the interleukin or cytokine is IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, GM-CSF, or G-CSF.