Claims for Patent: 9,821,114

Summary for Patent: 9,821,114

Title:	Compartmentalized method of nucleic acid delivery and compositions and uses thereof
Abstract:	Provided herein is a method of delivering nucleic acid molecules to a compartmentalized tissue or organ of a subject. Also provided herein are uses and processes for delivering a nucleic acid molecule to the parenchyma of a compartmentalized tissue or organ. The methods and uses can be used in the treatment of diseases and conditions and in industrial, agricultural and veterinary applications. Also provided herein are compositions containing an adenovirus or adeno-associated virus or other recombinant virus formulated for administration to the parenchyma of a tissue or organ.
Inventor(s):	Cabrera Aquino; Jose Gustavo (Mexico City, MX), Segura Pacheco; Blanca Angelica (Mexico City, MX)
Assignee:	Global Bio Therapeutics, Inc. (San Diego, CA)
Application Number:	13/815,206
Patent Claims:	1. A method of delivering a nucleic acid molecule to a subject, comprising: a) compartmentalizing a tissue or organ or portion of the tissue or organ from systemic circulation of the subject, wherein: the tissue or organ is the liver of the subject or a portion of the liver of a subject; the portion has a volume of at least 5 mm.sup.3; and compartmentalization includes clamping the tissue, organ or portion thereof, and effects isolation of the tissue, organ or portion from systemic circulation; b) administering a delivered agent directly to the parenchyma of the compartmentalized tissue or organ or portion of the tissue or organ, wherein: the delivered agent comprises the nucleic acid molecule, wherein, upon administration directly into the parenchyma, the delivered agent enters parenchymal cells of the compartmentalized tissue or organ or portion of the tissue or organ, and is not delivered into systemic circulation; and administration is effected by injection directly into the parenchyma in the compartmentalized tissue or organ or portion thereof, and does not include administration to connective tissue, blood vessels, nerves or ducts; c) maintaining the compartmentalization after administering the delivered agent for a predetermined time to effect cellular uptake of the delivered agent into cells of the parenchyma, wherein the predetermined time is at least 20 minutes up to 60 minutes; and d) after the predetermined time to effect cellular uptake of the delivered agent, restoring communication between the tissue, organ or portion of the tissue or organ and the systemic circulation. 2. The method of claim 1, wherein compartmentalizing the tissue or organ or portion of the tissue or organ is effected prior to or when administering the delivered agent. 3. The method of claim 1, wherein compartmentalizing the tissue or organ or portion of the tissue or organ is effected prior to administering the delivered agent, whereby the delivered agent is administered after compartmentalizing the tissue or organ or portion of the tissue or organ. 4. The method of claim 3, wherein administering the delivered agent is effected no more than 5 minutes after compartmentalizing the tissue or organ or portion of the tissue or organ. 5. The method of claim 1, wherein compartmentalization is achieved by applying pressure to the organ, tissue or portion thereof with a clamp, thereby compressing any arteries, veins, ducts and/or vessels and blocking the blood supply and flow to the arteries, veins, ducts and/or vessels in the tissue or organ or portion of the tissue or organ that is clamped, thereby eliminating the blood supply and flow to the tissue or organ or portion thereof that is clamped. 6. The method of claim 5, wherein all arteries, veins, ducts and/or vessels servicing or traversing the tissue or organ or portion of the tissue or organ are blocked from communication with the compartmentalized organ, tissue or portion thereof. 7. The method of claim 6, wherein the blocking of the arteries, veins, ducts and/or vessels is effected with a device or technique selected from among manual compression, an arterial or venous clamp, an occlusion catheter, a stapling device, a band, a tourniquet and a cable. 8. The method of claim 5, wherein a tissue or organ or portion of a tissue or organ is compartmentalized by clamping the tissue, organ or portion thereof with a parenchymal clamp. 9. The method of claim 8, wherein the clamp is a laparoscopic clamp. 10. The method of claim 1, wherein after compartmentalizing the tissue or organ or portion thereof, further comprising applying suction to reduce or eliminate blood in the tissue or organ or portion thereof. 11. The method of claim 1, wherein the predetermined time to effect cellular uptake is at least 30 minutes. 12. The method of claim 11, wherein restoring communication with the systemic circulation comprises removing the device or technique used to compartmentalize the tissue, organ or portion of the tissue or organ. 13. The method of claim 11, the method steps (a)-(d) are repeated a plurality of times. 14. The method of claim 13, wherein in a subsequent iteration of the method, the delivered agent is administered to the same compartmentalized locus or a different compartmentalized locus. 15. The method of claim 11, wherein the predetermined time is up to 45 minutes after administering the delivered agent. 16. The method of claim 11, wherein the predetermined time is less than 60 minutes. 17. The method of claim 11, wherein the predetermined time is 25 minutes to 60 minutes. 18. The method of claim 1, wherein the predetermined time is sufficient for the administered delivered agent to enter the parenchymal cells, whereby no more than 5% of the delivered agent is exposed to the systemic circulation. 19. The method of claim 18, wherein the predetermined time is up to 50 minutes after administering the delivered agent. 20. The method of claim 19, wherein the predetermined time is at least 30 minutes after administering the delivered agent. 21. The method of claim 1, wherein the tissue or organ or portion of the tissue or organ is a lobe of the liver or portion thereof. 22. The method of claim 1, wherein the portion of the liver is an area of that has a length ranging from about 0.5 cm to 25 cm, a height of 0.5 cm to 20 cm and a depth from 0.5 cm to 15 cm. 23. The method of claim 1, wherein: the predetermined time before restoring communication with the systemic circulation is between about 20 minutes and 50 minutes. 24. The method of claim 1, wherein a portion of the tissue or organ is compartmentalized. 25. The method of claim 1, wherein a portion of the liver is compartmentalized and the portion is a lobe, segment or a portion of a lobe or segment of the liver. 26. The method of claim 25, wherein the lobe or portion of a lobe is selected from among the right lobe, the left lobe, the quadrate lobe and the caudate lobe or is a portion thereof. 27. The method of claim 1, wherein the delivered agent is selected from among a non-viral vector, a virus, a virus-like particle, a minicircle, a nanoparticle and a whole cell that comprise the nucleic acid molecule. 28. The method of claim 27, wherein the delivered agent is a nanoparticle and the nanoparticle is targeted or radiolabeled. 29. The method of claim 27, wherein the delivered agent is a virus and the virus is selected from among an adenovirus, an adeno-associated virus (AAV), a retrovirus, vaccinia virus and herpes simplex virus. 30. The method of claim 29, wherein the retrovirus is a lentivirus. 31. The method of claim 29, wherein the virus is an adenovirus. 32. The method of claim 31, wherein the adenovirus comprises a deletion in an E1, E2a, E2b, E3, or E4 coding region. 33. The method of claim 31, wherein the adenovirus is adenovirus type 2 or adenovirus type 5. 34. The method of claim 33, wherein the adenovirus comprises a deletion in an E1, E2a, E2b, E3, or E4 coding region. 35. The method of claim 29, wherein the virus comprises the nucleic acid molecule and the nucleic acid molecule is heterologous to the virus genome. 36. The method of claim 1, wherein the nucleic acid molecule encodes a polypeptide, whereby the polypeptide is expressed. 37. The method of claim 36, wherein the encoded polypeptide is selected from the among an enzyme, a hormone, a coagulation or clotting factor, a cytokine, a growth factor or active portion thereof, an antibody or antigen binding portions of antibodies, an angiogenesis modulator, an immunomodulator, a pain modulator, a receptor or active portion thereof, a transport protein, a regulatory protein, an antigen and an allergen. 38. The method of claim 36, wherein the encoded polypeptide is selected from among adenosine deaminase, cystic fibrosis transmembrane conductance regulator (CTFR), galsulfase, laronidase, N-acetylgalactosamine 6-sulfatase, phenylalanine ammonia lyase, acid alpha glucosidase, imiglucerase, alglucosidase alpha, thyrotropin, growth hormone, insulin, thyroid hormone, erythropoietin (EPO), interleukin-1 (IL-1), IL-2, IL-3, IL-4, IL-5, IL-7, interferon-.alpha. (IFN-.alpha.), IFN-.beta., IFN-.gamma., tumor necrosis factor (TNF), IL-12, IL-18, fms-related tyrosine kinase 3 (flt3), neuropilin-2 (NP2), bone morphogenic protein (BMPs), epidermal growth factor (EGF), erythropoietin (EPO), fibroblast growth factor (FGF), granulocyte colony stimulating factor (G-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), platelet derived growth factor (PDGF), transforming growth factor .alpha. or .beta., vascular endothelial growth factor (VEGF), epidermal growth factor receptor (EGFR), fibroblast growth factor receptor (FGFR), FGFR antagonist (sFGFR) transforming growth factor receptor (TGFR), vascular endothelial growth factor receptor (VEGFR), plasminogen activator, urokinase, Factor VIII, Factor IX, von Willebrand factor, growth hormone, metalloproteinase thrombospondin motifs 1 (METH-1), METH-2, tryptophanyl-tRNA synthetase (TrpRS) fragments, proliferin-related protein, prolactin fragment, pigment epithelium-derived factor (PEDF), vasostatin, angiostatin, endostatin, kininostatin, fibrinogen-E fragment, thrombospondin, tumstatin, canstatin, restin, soluble fms-like tyrosine kinase-1 (sFlt-1), soluble vascular endothelial growth factor receptors (sFlk), soluble Neuropilin 1 (sNRP1), Interferon gamma-induced protein 10 (IP-10), Platelet factor 4 (PF-4), Gro-beta, soluble Ephrin type-B receptor 4 (sEphB4), sephrinB2, IGF-1, herpes simplex virus thymidine kinase (HSV-TK), carboxypeptidase G2 (CPG2), carboxylesterase (CA), cytosine deaminase (CD), cytochrome P450 (cyt-450), deoxycytidine kinase (dCK), nitroreductase (NR), purine nucleoside phosphorylase (PNP), thymidine phosphorylase (TP), varicella zoster virus thymidine kinase (VZV-TK), xanthine-guanine phosphoribosyl transferase (XGPRT), Aspartylglucosaminidase, .alpha.-Galactosidase A, Palmitoyl Protein Thioesterase, Tripeptidyl Peptidase, Lysosomal transmembrane protein, cysteine transporter, Acid ceramidase, acid .alpha.-L-fucosidase, protective protein/cathepsin A, acid .beta.-glucosidase or glucocerebrosidase, acid .beta.-galactosidase, iduronate-2-sulfatase, .alpha.-L-Iduronidase, galactocerebrosidase, acid .alpha.-mannosidase, acid .beta.-mannosidase, arylsulfatase B, arylsulfatase A, N-Acetylgalactosamine-6-sulfate sulfatase, N-Acetlylglucosamine-1-phosphotransferase, Acid sphingomyelinase, NPC-1, .beta.-Hexosaminidase B, Heparan N-sulfatase, .alpha.-N-Acetylglucosaminidase (NaGlu), Acetyl-CoA:.alpha.glucosamininde N-acetyltransferase, N-Acetylglucosamine-6-sulfate sulfatase, .beta.-Glucuronidase, acid lipase, neprilysin, the insulin-degrading enzyme insulysin, thimet oligopeptidase, calbindin D28, parvalbumin, HIF1-alpha, SIRT-2, SMN-1, SMN-2, GDNF, ciliary neurotrophic factor (CNF), low density lipoprotein receptor (LDLR), lipoprotein lipase (LPL), Alpha-1-Antitrypsin (AAT), UDP-glucuronyl-transferase (UGT), UGT1A1, glucose-6 phosphatase, phosphoenolpyruvate-carboxykinase, galactose-1 phosphate uridyl transferase, phenylalanine hydroxylase, branched chain alpha-ketoacid dehydrogenase, fumarylacetoacetate hydrolase, methylmalonyl-CoA mutase, ornithine transcarbamylase, argininosuccinic acid synthetase, adenosine deaminase, hyposanthine guanine phosphoribosyl transferase, biotinidase, beta-glucocerebrosidase, beta-gluronidase, porphobilinogen deaminase (PBDG) and p53. 39. The method of claim 36, wherein the nucleic acid molecule is a therapeutic nucleic acid molecule that encodes a therapeutic product. 40. The method of claim 39, wherein the disease or condition is selected from among an arthritis, chronic pain, HIV-related AIDS, atherosclerosis, restenosis, inherited enzyme deficiency, inherited immune deficiency, cancer, a retrovirus infection, hemophilia, diabetes, a muscular dystrophy, a cardiovascular disorder, cystic fibrosis, a neurodegenerative disorder, trauma, pain, sickle cell anemia, autoimmune disease, inflammatory disease, and hypertension. 41. The method of claim 39, wherein the nucleic acid molecule encode a protein selected from among a Factor VIII for the treatment of hemophilia A; a Factor IX for the treatment of hemophilia B; an insulin gene for treatment of type I diabetes mellitus; an alpha-1-antitrypsin (AAT) for the treatment of alpha-1-antitrypsin (AAT) deficiency; a hemochromatosis protein (HFE) for treatment of hemochromatosis; a copper-transporting ATPase 2 for treatment of Wilson's disease; UDP glucuronosyltransferase 1A1 (UGT1A1) for the treatment of Crigler-Najjar syndrome type I; ornithine transcarbamylase (OTC) for the treatment of ornithine transcarbamylase deficiency, type II; low density lipoprotein receptor (LDLR) for the treatment of familial hypercholesterolemia; fibrinogen alpha (FGA), beta (FGB) or gamma (FGB) for the treatment of afibrinogenemia; glucose-6-phosphate-.alpha. for the treatment of glycogen storage disease (GSD) type Ia; G6PT for the treatment of GSD type Ib; acid-.alpha.-glucosidase for the treatment of GSD type II (Pompe); .alpha.-L-iduronidase for the treatment of mucopolysaccharidosis (MPSI); sulfamidase for the treatment of MPS IIIA; .alpha.-N-acetylglucosaminidase (NaGlu) for the treatment of MPS IIIB; .beta.-glucuronidase for the treatment of MPS VII; .alpha.-galactosidase A for the treatment of Fabry disease; glucocerebrosidase for the treatment of Gaucher's disease; acid sphingomyelinase for the treatment of Niemann-Pick syndrome; phenylalanine hydroxylase for the treatment of phenylketonuria; TIMP antagonist or anti-HSC molecules for the treatment of liver fibrosis; anti-ROS molecules for the treatment of liver ischemia reperfusion injury; amyloid-beta degrading enzyme neprilysin, the insulin-degrading enzyme insulysin, or thimet oligopeptidase for the treatment of Alzheimer's disease; insulin growth factor-1 (IGF-1), calbindin D28, parvalbumin, HIF1-alpha, SIRT-2, VEGF, SMN-1, SMN-2, GDNF or ciliary neurotrophic factor (CNF) for the treatment of Amyotrophic Lateral Sclerosis (ALS); galactose-1 phosphate uridyl transferase for the treatment of galactosemia; branched chain alpha-ketoacid dehydrogenase for the treatment of maple syrup urine disease; fumarylacetoacetate hydrolase for the treatment of tyrosinemia type 1; methylmalonyl-CoA mutase for the treatment of methylmalonic acidemia; argininosuccinic acid synthetase for the treatment of citrullinemia; hyposanthine guanine phosphoribosyl transferase for the treatment of Gout and Lesch Nyan syndrome; beta-gluronidase for the treatment of Sly syndrome; peroxisome membrane protein 70 kDa for the treatment of Zellweger syndrome, enfuvirtide for the treatment of Human immunodeficiency virus (HIV) infection; adenosine deaminase (ADA) for the treatment of combined immunodeficiency disease (SCID); CFTR for the treatment of cystic fibrosis; porphobilinogen deaminase (PBDG) for the treatment of acute intermittent porphyria; interferon-beta for the treatment of multiple sclerosis; lipoprotein lipase for the treatment of lipoprotein lipase deficiency (LPLD), p53 for the treatment of cancer; and glutamic acid decarboxylase (GAD) for the treatment of Parkinson's Disease. 42. The method of claim 36, wherein the nucleic acid molecule encodes a polypeptide that increases muscle production in an animal, increases hair production in an animal, increases wool production in an animal, increases growth of an animal, or is involved in nutrient synthesis or utilization. 43. The method of claim 42, wherein the encoded polypeptide is selected from among: a polypeptide that increases muscle production in an animal that is a myostatin inhibitor; a polypeptide that increases growth in an animal that is a growth hormone, IGF-1, a growth hormone releasing factor or chicken Ski; and a polypeptide that is involved in nutrient synthesis or utilization that is a serine transacetylase and o-acetylserine sulphydrylase. 44. The method of claim 43, wherein the myostatin inhibitor is follistatin. 45. The method of claim 1, wherein the nucleic acid molecule is selected from among a DNA molecule, a RNA molecule, and an aptamer. 46. The method of claim 45, wherein the nucleic acid molecule is selected from among a microRNA, a small interfering RNA, a ribozyme and an antisense nucleic acid. 47. The method of claim 1, comprising imaging the tissue or organ to identify the parenchymal tissue prior to administration of the delivered agent. 48. The method of claim 47, wherein the imaging is magnetic resonance imaging (MRI), sonography (ultrasound), or computed tomography (ct). 49. The method of claim 1, wherein the delivered agent is formulated with lipids, polymer reagents or other agents to facilitate entry into the parenchymal cells. 50. The method of claim 1, wherein the delivered agent is delivered in the presence of a physical method to facilitate entry into parenchymal cells. 51. The method of claim 50, wherein the physical method is selected from among electroporation, sonoporation, hydrodynamic pressure, ultrasound and a gene gun. 52. The method of claim 1, further comprising administering an agent that promotes cellular uptake of the delivered agent, wherein the agent is administered prior to, simultaneously or subsequent to administration of the delivered agent. 53. The method of claim 52, wherein the agent is a transcriptional enhancer of a virus-specific cell surface receptor or of the therapeutic transgene. 54. The method of claim 53, wherein the agent is a histone deacetylase (HDAC) inhibitor. 55. The method of claim 1, wherein the amount of delivered agent that is administered is 100-fold or less than the amount of the same delivered agent administered intravenously for the same purpose. 56. The method of claim 1, wherein: the delivered agent is an adenovirus or adeno-associated virus comprising a heterologous nucleic acid molecule in its genome; and the amount of delivered agent administered is between about 10 to 1.times.10.sup.12 viral particles. 57. The method of claim 56, wherein: the amount of delivered agent administered is up to about 1.times.10.sup.10 to about 1.times.10.sup.11 viral particles. 58. The method of claim 56, wherein: the amount of delivered agent administered is up to about 1.times.10.sup.7 viral particles. 59. The method of claim 1, wherein the delivered agent is administered to more than one locus in the compartmentalized tissue, organ or portion of the tissue or organ. 60. The method of claim 1, wherein prior to restoring communication to the tissue, organ or portion of the tissue or organ with the systemic circulation, further comprising removing from the parenchyma of the tissue or organ or portion of the tissue or organ any delivered agent that is extracellular. 61. The method of claim 1, wherein the subject is selected from among a mouse, rat, cow, pig, sheep, goat, horse and human. 62. The method of claim 1, wherein the subject is a human child under the age of 18 or is a human fetus. 63. The method of claim 1, wherein the subject has been diagnosed with a genetic deficiency. 64. The method of claim 1 the method steps (a)-(d) are performed via laparoscopy. 65. The method of claim 1, wherein the predetermined time is no more than 30 minutes after administering the delivered agent. 66. The method of claim 1, wherein the portion has a volume of at least 10 mm.sup.3. 67. The method of claim 1, wherein the predetermined time is 20 minutes to 30 minutes. 68. The method of claim 1, wherein the nucleic acid encodes insulin. 69. The method of claim 1, wherein the nucleic acid encodes a coagulation or clotting factor. 70. The method of claim 1, wherein the nucleic acid encodes factor VIII. 71. The method of claim 1, wherein the predetermined time is 30 to 60 minutes after compartmentalization. 72. The method of claim 1, wherein the predetermined time is at least 25 minutes after compartmentalization. 73. A method of delivering a nucleic acid molecule to a subject, comprising: a) compartmentalizing a tissue or organ or portion of the tissue or organ to isolate it from systemic circulation of the subject, wherein: the portion has a volume of at least 5 mm.sup.3; and compartmentalization is effected by clamping the tissue or organ or portion of the tissue or organ to effect isolation of the tissue, organ or portion from systemic circulation; b) administering a delivered agent directly by injection into the parenchyma of the compartmentalized tissue or organ or portion of the tissue or organ, wherein: the delivered agent comprises the nucleic acid molecule, whereby the delivered agent directly enters parenchymal cells of the compartmentalized tissue or organ or portion of the tissue or organ; and the parenchyma in the compartmentalized tissue or organ or portion of the tissue or organ does not include connective tissue, blood vessels, nerves or ducts; and c) maintaining the compartmentalization for a predetermined time to effect cellular uptake of the delivered agent in to the cells of the parenchyma, wherein the predetermined time is more than 15 minutes up to about 1 hour; and d) after the predetermined time to effect cellular uptake of the delivered agent, restoring communication between the tissue, organ or portion of the tissue or organ and the systemic circulation. 74. The method of claim 73, wherein the predetermined time is at least 30 minutes. 75. The method of claim 73, wherein the predetermined time is at least 25 minutes after compartmentalization.

Details for Patent 9,821,114

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genzyme Corporation	CEREZYME	imiglucerase	For Injection	020367	05/23/1994		2032-02-07
Genzyme Corporation	CEREZYME	imiglucerase	For Injection	020367	09/22/1999		2032-02-07
Microbix Biosystems Inc.	KINLYTIC	urokinase	For Injection	021846	01/16/1978		2032-02-07
Biomarin Pharmaceutical Inc.	ALDURAZYME	laronidase	Injection	125058	04/30/2003		2032-02-07
Biomarin Pharmaceutical Inc.	NAGLAZYME	galsulfase	Injection	125117	05/31/2005		2032-02-07