Tuneable delivery of nanoparticle bound active plasmin for the treatment of thrombosis

Abstract

Compositions and methods for therapeutic delivery are disclosed. More particularly, the present disclosure relates to nanoparticle compositions that sequester the activity of a target molecule while leaving other domains accessible to bind targeted tissues of interest. Methods for thrombus dissolution include administering a nanoparticle reversibly coupled to a target molecule that can dissolve a blood clot. Compositions and methods for inducing blood clotting are also disclosed. Methods for inducing blood clotting include administering a nanoparticle reversibly coupled to a target molecule that can induce the formation of a blood clot. Methods for sequestering a target molecule are also disclosed. The method includes reversibly coupling a target molecule to a nanoparticle having an affinity ligand that reversibly couples the target molecule, and thus, sequesters the target molecule activity until the target molecule interacts with its substrate resulting in the release of the target molecule.

1 .- 27 . (canceled) 
     
     
         28 . A method of sequestering an enzyme in vivo in a subject in need thereof, the method comprising:
 administering to the subject a nanoparticle that comprises an affinity ligand covalently coupled to an exterior surface of the nanoparticle, wherein the affinity ligand can reversibly couple the enzyme.   
     
     
         29 . The method of  claim 28 , wherein the nanoparticle is selected from the group consisting of a micelle, a liposome, a dendrimer, a biodegradable polymer scaffold, a non-biodegradable polymer scaffold, an inorganic nanoparticle, and combinations thereof. 
     
     
         30 . The method of  claim 28 , wherein the lipid molecule is selected from the group consisting of phosphatidylcholine, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, a phosphoinositide, a phosphingolipid and combinations thereof. 
     
     
         31 . The method of  claim 28 , wherein the affinity ligand is selected from the group consisting of a small molecule, a peptide, a peptidomimetic and combinations thereof. 
     
     
         32 . The method of  claim 31 , wherein the affinity ligand is selected from the group consisting of a benzamidine, bivalirudin, argatroban, melagatran, ximelagatran, dabigatran, amprenavir, atazanavir, darunavir, fosamprenavir, indinavir, lopinavir/ritonavir combination, nelfinavir, ritonavir, saquinavir, tipranavir, and combinations thereof. 
     
     
         33 . The method of  claim 31 , wherein the affinity ligand is selected from the group consisting of a serpin, hirudin, bivalirudin, lepirudin, desirudin, and combinations thereof. 
     
     
         34 . The method of  claim 28 , wherein the nanoparticle further comprises a linker. 
     
     
         35 . The method of  claim 28 , wherein the nanoparticle further comprises cholesterol. 
     
     
         36 . The method of  claim 28 , wherein the enzyme is selected from the group consisting of an oxidoreductase, a transferase, a hydrolase, a lyase, an isomerase, a ligase, and combinations thereof. 
     
     
         37 . The method of  claim 36 , wherein the oxidoreductase is selected from the group consisting of alcohol dehydrogenase (NAD), alcohol dehydrogenase (NADP), homoserine dehydrogenase, aminopropanol oxidoreductase, diacetyl reductase, glycerol dehydrogenase, propanediol-phosphate dehydrogenase, glycerol-3-phosphate dehydrogenase, D-xylulose reductase, L-xylulose reductase, lactate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, HMG-CoA reductase, glucose oxidase, L-gulonolactone oxidase, thiamine oxidase, xanthine oxidase, acetaldehyde dehydrogenase, Glyceraldehyde 3-phosphate dehydrogenase, Pyruvate dehydrogenase, Oxoglutarate dehydrogenase, Biliverdin reductase, Protoporphyrinogen oxidase, monoamine oxidase, dihydrofolate reductase, methylenetetrahydrofolate reductase, sarcosine, oxidase, Dihydrobenzophenanthridine oxidase, NADH dehydrogenase, urate oxidase, nitrite reductase, nitrate reductase, glutathione reductase, thioredoxin reductase, sulfite oxidase, cytochrome C oxidase, coenzyme Q (cytochrome C reductase), catechol oxidase, laccase, cytochrome C peroxidase, catalase, myeloperoxidase, thyroid peroxidase, glutathione peroxidase, 4-hydroxyphenylpyruvate dioxygenase, cytochrome P450 oxidase, aromatase, CYP3D6, CYP2E1, CYP3A4, nitric oxide dioxygenase, nitric oxide synthase, aromatase, CUP2D6, CYP2E1, CYP3A4, phenylalanine hydroxylase, tyrosinase, superoxide dismutase, ceruloplasmin, leucoanthocyanidin reductase, xanthine dehydrogenase, nicotinate dehydrogenase, 4-hydroxy-tetrahydrodipicolinate reductase, Nicotinate dehydrogenase (cytochrome), xanthine oxidase, ribonucleotide reductase, Ribonucleoside-triphosphate reductase, Vitamin K epoxide reductase, Vitamin-K-epoxide reductase (warfarin-sensitive), Vitamin-K-epoxide reductase (warfarin-insensitive), RRM1, RRM2, RRM2B, caffeine dehydrogenase, nitrogenase, nitrogenase (flavodoxin), arsenate reductase (glutaredoxin), glutaredoxin, iodotyrosine deiodinase, Isopenicillin N synthase, Tetrahydrocannabinolic acid synthase, Thyroxine 5-deiodinase, Iodothyronine deiodinase, Deiodinase, and combinations thereof. 
     
     
         38 . The method of  claim 36 , wherein the transferase is selected from the group consisting of Glutathione S-transferase, Catechol-O-methyl transferase, DNA methyltransferase, Histone methyltransferase, Aspartate transcarbamoylase, Ornithine transcarbamoylase, Transketolase, Transaldolase, Acetolactate synthase, 2-Succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylic-acid synthase, Aminolevulinic acid synthase, Choline acetyltransferase, Factor XIII, Gamma glutamyl transpeptidase, Transglutaminase, Hypoxanthine-guanine phosphoribosyltransferase, Thiaminase, Flavin prenyltransferase, Alanine transaminase, Aspartate transaminase, Butyrate kinase, Thiosulfate sulfurtransferase, 3-mercaptopyruvate sulfurtransferase, Thiosulfate—thiol sulfurtransferase, tRNA uracil 4-sulfurtransferase, Thiosulfate—dithiol sulfurtransferase, Biotin synthase, Cysteine desulfurase, Lipoyl synthase, Molybdenum cofactor sulfurtransferase, Thiazole synthase, Molybdopterin synthase sulfurtransferase, Molybdopterin synthase, tRNA-uridine 2-sulfurtransferase, tRNA-5-taurinomethyluridine 2-sulfurtransferase, tRNA-5-methyluridine(54) 2-sulfurtransferase, L-seryl-tRNA(Sec) selenium transferase, O-phospho-L-seryl-tRNA(Sec):L-selenocysteinyl-tRNA synthase, and combinations thereof. 
     
     
         39 . The method of  claim 36 , wherein the hydrolase is selected from the group consisting of hydrolytic enzyme, nuclease, endonuclease, exonuclease, acid hydrolase, phospholipase A, acetylcholinesterase, cholinesterase, lipoprotein lipase, Ubiquitin carboxy-terminal hydrolase L1, phosphatase, alkaline phosphatase, fructose bisphosphatase, phospholipase C, CGMP specific phosphodiesterase type 5, phospholipase D, restriction enzyme Type 1, restriction enzyme Type 2, restriction enzyme Type 3, restriction enzyme Type 4, deoxyribonuclease, RNase H, ribonuclease, amylase, sucrase, chitinase, lysozyme, maltase, lactase, beta-galactosidase, hyaluronidase, Adenosylmethionine hydrolase, S-adenosyl-L-homocysteine hydrolase, Alkenylglycerophosphocholine hydrolase, Alkenylglycerophosphoethanolamine hydrolase, Cholesterol-5,6-oxide hydrolase, Hepoxilin-epoxide hydrolase, Isochorismatase, Leukotriene-A4 hydrolase, Limonene-1,2-epoxide hydrolase, Microsomal epoxide hydrolase, Trans-epoxysuccinate hydrolase, Alanine aminopeptidase, Angiotensin converting enzyme, serine protease, chymotrypsin, trypsin, thrombin, factor X, plasmin, acrosin, factor VII, factor IX, prolyl oligopeptidase, factor XI, elastase, factor XII, proteinase K, tissue plasminogen activator, protein C, separase, pepsin, rennet, renin, trypsinogen, plasmepsin, matrix metalloproteinase, metalloendopeptidase, urease, beta-lactamase, arginase, adenosine deaminase, GTP cyclohydrolase I, nitrilase, helicase, DnaB helicase, RecQ helicase, ATPase, NaKATPase, ATP synthase, kynureninase, haloacetate dehalogenase, phosphoamidase, protein arginine phosphatase, phosphohistidine phosphatase, N-sulfoglucosamine sulfohydrolase, Cyclamate sulfohydrolase, Phosphonoacetaldehyde hydrolase, Phosphonoacetate hydrolase, Phosphonopyruvate hydrolase, Trithionate hydrolase, UDP-sulfoquinovose synthase, 2′-hydroxybiphenyl-2-sulfinate desulfinase, 3-sulfinopropanoyl—CoA desulfinase, Carbon disulfide hydrolase, (CysO sulfur-carrier protein)-S-L-cysteine hydrolase, Carbonyl sulfide hydrolase, S-adenosyl-L-methionine hydrolase (adenosine-forming), and combinations thereof. 
     
     
         40 . The method of  claim 36 , wherein the lyase is selected from the group consisting of Ornithine decarboxylase, Uridine monophosphate synthetase, Aromatic-L-amino-acid decarboxylase, RubisCO, Fructose-bisphosphate aldolase, Carbonic anhydrase, Tryptophan synthase, Phenylalanine ammonia-lyase, Cystathionine gamma-lyase, Cystathionine beta-lyase, Leukotriene C4 synthase, Dichloromethane dehalogenase, Halohydrin dehalogenase, Adenylate cyclase, Guanylate cyclase, and combinations thereof. 
     
     
         41 . The method of  claim 36 , wherein the isomerase is selected from the group consisting of Amino-acid racemase, Serine racemase, Mandelate racemase, UDP-glucose 4-epimerase, Methylmalonyl CoA epimerase, FKBP (FKBP1A), FKBP1B, FKBP2, FKBP3, FKBP4, FKBP5, FKBP6, FKBP7, FKBP8, FKBP9, FKBP10, FKBPL, Cyclophilin, Prolyl isomerase, 2-chloro-4-carboxymethylenebut-2-en-1,4-olide isomerase, Beta-carotene isomerase, Farnesol 2-isomerase, Furylfuramide isomerase, Linoleate isomerase, Maleate isomerase, Maleylacetoacetate isomerase, Maleylpyruvate isomerase, Photoisomerase, Prolycopene isomerase, Prolyl isomerase, Retinal isomerase, Retinol isomerase, Zeta-carotene isomerase, Enoyl CoA isomerase, Protein disulfide isomerase, Phosphoglucomutase, Muconate cycloisomerase, 3-carboxy-cis,cis-muconate cycloisomerase, Tetrahydroxypteridine cycloisomerase, Inositol-3-phosphate synthase, Carboxy-cis,cis-muconate cyclase, Chalcone isomerase, Chloromuconate cycloisomerase, (+)-bornyl diphosphate synthase, Cycloeucalenol cycloisomerase, Alpha-pinene-oxide decyclase, Dichloromuconate cycloisomerase, Copalyl diphosphate synthase, Ent-copalyl diphosphate synthase, Syn-copalyl-diphosphate synthase, Terpentedienyl-diphosphate synthase, Halimadienyl-diphosphate synthase, (S)-beta-macrocarpene synthase, Lycopene epsilon-cyclase, Lycopene beta-cyclase, Prosolanapyrone-III cycloisomerase, D-ribose pyranase, Steroid Delta Isomerase, Topoisomerase, and combinations thereof. 
     
     
         42 . The method of  claim 36 , wherein the ligase is selected from the group consisting of 6-carboxytetrahydropterin synthase, Phenylalanyl-tRNA synthetase beta chain, Acetate—CoA ligase, Medium-chain acyl—CoA ligase, Long-chain-fatty-acid—CoA ligase, Succinate—CoA ligase (GDP-forming), Succinate—CoA ligase (ADP-forming), Glutarate—CoA ligase, Cholate—CoA ligase, Oxalate—CoA ligase, Malate—CoA ligase, Acid—CoA ligase (GDP-forming), Biotin—CoA ligase, 4-coumarate—CoA ligase, Acetate—CoA ligase (ADP-forming), 6-carboxyhexanoate—CoA ligase, Arachidonate—CoA ligase, Acetoacetate—CoA ligase, Propionate—CoA ligase, Citrate—CoA ligase, Long-chain-fatty-acid-luciferin-component ligase, Long-chain-fatty-acid-(acyl-carrier-protein) ligase, (citrate (pro-3S)-lyase) ligase, Dicarboxylate—CoA ligase, Phytanate—CoA ligase, Benzoate—CoA ligase, o-succinylbenzoate—CoA ligase, 4-hydroxybenzoate—CoA ligase, 3-alpha,7-alpha-dihydroxy-5-beta-cholestanate—CoA ligase, Phenylacetate—CoA ligase, 2-furoate—CoA ligase, Anthranilate—CoA ligase, 4-chlorobenzoate—CoA ligase, Trans-feruloyl-CoA synthase, ACP-SH:acetate ligase, 3-hydroxypropionyl-CoA synthase, 3-hydroxybenzoate—CoA ligase, (2,2,3-trimethyl-5-oxocyclopent-3-enyl)acetyl-CoA synthase, (butirosin acyl-carrier protein)-L-glutamate ligase, 4-hydroxybutyrate-CoA ligase, 3-((3aS,4S,7aS)-7a-methyl-1,5-dioxo-octahydro-1H-inden-4-yl)propanoate-CoA ligase, 3-oxocholest-4-en-26-oate-CoA ligase, 2-hydroxy-7-methoxy-5-methyl-1-naphthoate-CoA ligase, 3-(methylthio)propionyl-CoA ligase, E1 ubiquitin-activating enzyme, L-allo-isoleucine-holo-CmaA peptidyl-carrier protein ligase, Medium-chain-fatty-acid-(acyl-carrier-protein) ligase, Carnitine-CoA ligase, Long-chain fatty acid adenylyltransferase FadD28, 4-hydroxybenzoate adenylyltransferase FadD22, 4-hydroxyphenylalkanoate adenylyltransferase FadD29, L-proline-L-prolyl-carrier protein ligase, D-alanine-D-alanyl-carrier protein ligase, E1 SAMP-activating enzyme, Glutamine synthetase, Argininosuccinate synthetase, CTP synthase, Pyruvate carboxylase, Acetyl-CoA carboxylase, DNA ligase, and combinations thereof. 
     
     
         43 . The method of  claim 28 , wherein the enzyme is a coagulation factor. 
     
     
         44 . The method of  claim 43 , wherein the coagulation factor is selected from the group consisting of an antifibrinolytic, a blood coagulation factor, fibrinogen, prothrombin, thrombin, tissue factor, tissue factor thromboplastin, factor IV, factor VI, factor XII, factor XI (plasma thromboplastin antecedent (PTA), antihemophilic factor C), factor XIa, factor XII, factor XIIa, factor X (Stuart-Prower factor, Stuart factor), prothrombin, thrombin, a thrombin like enzyme, batroboxin (reptilase), tissue plasminogen activator, plasminogen, protein C, protein S, protein Z, proaccelerin, labile factor, Ac-globulin, collagen, vitamin K, chitosan, von Willebrand factor, stable factor, proconvertin, serum prothrombin conversion accelerator (SPCA), antihemophilic factor A, antihemophilic factor (AHF), antihemophilic globulin (AHG), antihemophilic factor B, Christmas factor, plasma thromboplastin component (PTC), Prekallikrein, Kallikrein, High-molecular-weight kininogen (HMWK) (Fitzgerald factor), Antithrombin III, Heparin cofactor II, Protein Z-related protease inhibitor (ZPI), α2-Antiplasmin, α2-Macroglobulin, Urokinase, Plasminogen activator inhibitor-1 (PAI-1), Plasminogen activator inhibitor-2 (PAI-2), Cancer procoagulant, and combinations thereof. 
     
     
         45 . The method of  claim 28 , wherein the subject has or is suspected of having a bleeding disorder. 
     
     
         46 . The method of  claim 39 , wherein the bleeding disorder is selected from the group consisting of coronary thrombosis, cerebrovascular thrombosis, deep vein thrombosis, pulmonary thrombosis, hemophilia, and Von Willebrand disease.