Pharmacological vitreolysis

Abstract

A method of treating or preventing a disorder, or a complication of a disorder, of an eye of a subject comprising contacting a vitreous and/or aqueous humor with a composition comprising a truncated form of plasmin comprising a catalytic domain of plasmin (TPCD). TPCDs include, but are not limited to, miniplasmin, microplasmin and derivatives and variants thereof. The methods of the invention can be used to reduce the viscosity of the vitreous, liquefy the vitreous, induce posterior vitreous detachment, reduce hemorrhagic blood from the eye, clear or reduce materials toxic to the eye, clear or reduce intraocular foreign substances from the eye, increase diffusion of a composition administered to an eye, reduce extraretinal neovascularization and any combinations thereof. The method can be used in the absence of, or as an adjunct to, vitrectomy.

1 . A method of treating a vitreoretinal disease or disorder, or of treating a complication of a vitreoretinal disease or disorder, of an eye of a subject, comprising contacting the aqueous humor in the eye of the subject with an effective amount of a microplasmin and with an effective amount of a second agent, thereby treating the vitreoretinal disease or disorder, or the complication of the vitreoretinal disease or disorder, of the eye of the subject. 
     
     
         2 . The method according to  claim 1  wherein said aqueous humor is contacted with the second agent prior to, at the same time as, or after being contacted with said microplasmin. 
     
     
         3 . The method according to  claim 1  wherein the second agent is a protein other than said microplasmin, a chemical, or another substance useful in treating or preventing an eye disorder or a complication of an eye disorder. 
     
     
         4 . The method according to  claim 1  wherein said second agent is selected from the group consisting of hyaluronidase, chondroitinase ABC, chondroitinase AC, chondroitinase B, chondroitin 4-sulfatase, chondroitin 6-sulfatase, β-glucuronidase, collagenase, dispase, RGD containing peptides, echistatin, falvoridin, anti-integrin antibody, P2Y receptor antagonists, urea, hydroxyurea, thiourea, anti-angiogenic agents, VEGF inhibitors, PlGF inhibitors, and any combinations thereof. 
     
     
         5 . The method according to  claim 4  wherein said VEGF inhibitor is an anti-VEGF antibody, a VEGF-aptamer, or a soluble VEGF receptor. 
     
     
         6 . The method according to  claim 4  wherein said PlGF inhibitor is an anti-PlGF antibody, a PlGF-aptamer, or a soluble VEGF receptor. 
     
     
         7 . The method according to  claim 1  wherein said vitreoretinal disease or disorder is selected from the group consisting of retinal detachment, retinal tear, retinal neovascularization, vitreous hemorrhage, diabetic vitreous hemorrhage, proliferative diabetic retinopathy, non-proliferative diabetic retinopathy, macular pucker, macular exudates, fibrin deposition, retinal vein occlusion, retinal artery occlusion, subretinal hemorrhage, amblyopia, endophthalmitis, retinopathy of prematurity, glaucoma, retinitis pigmentosa, and any combination thereof. 
     
     
         8 . The method according to  claim 1  wherein said vitreoretinal disease or disorder is macular edema. 
     
     
         9 . The method according to  claim 8  wherein said macular edema is cystoid macular edema. 
     
     
         10 . The method according to  claim 1  wherein said vitreoretinal disease or disorder is age-related macular degeneration. 
     
     
         11 . The method according to  claim 1  wherein said vitreoretinal disease or disorder is macular hole. 
     
     
         12 . The method according to  claim 11  wherein said macular hole is selected from the group consisting of impending macular hole, full-thickness macular hole, idiopathic macular hole and traumatic macular hole. 
     
     
         13 . The method according to  claim 1  wherein said vitreoretinal disease or disorder is vitreomacular traction. 
     
     
         14 . The method according to  claim 1  wherein said vitreoretinal disease or disorder is incomplete posterior vitreous detachment. 
     
     
         15 . The method according to  claim 1  wherein said vitreoretinal disease or disorder is associated with vitreous contraction. 
     
     
         16 . The method according to  claim 1  wherein said microplasmin is selected from the group consisting of recombinant microplasmin, stabilized microplasmin, and stabilized, recombinant microplasmin. 
     
     
         17 . The method according to  claim 16  wherein said microplasmin is stabilized by contacting with a stabilizing agent. 
     
     
         18 . The method according to  claim 17  wherein said stabilizing agent is selected from the group consisting of tranexamic acid, hexanoic acid, lysine, serine, threonine, methionine, glutamine, alanine, glycine, isoleucine, valine, alanine aspartic acid, polyhydric alcohol, a pharmaceutically acceptable carbohydrate, glucosamine, thiamine, niacinamide, an acidic buffer, a salt, and any combination thereof. 
     
     
         19 . The method according to  claim 18  wherein said acidic buffer is comprising acetic acid, benzoic acid, carboxylic acid, citric acid, hydrochloric acid, lactic acid, malic acid or tartaric acid. 
     
     
         20 . The method according to  claim 18  wherein said salt is calcium chloride, magnesium chloride, potassium chloride or sodium chloride. 
     
     
         21 . The method according to  claim 16  wherein said microplasmin is purified in the presence of a stabilizing agent. 
     
     
         22 . The method according to  claim 21  wherein said stabilizing agent is selected from the group consisting of tranexamic acid, hexanoic acid, lysine, serine, threonine, methionine, glutamine, alanine, glycine, isoleucine, valine, alanine aspartic acid, polyhydric alcohol, a pharmaceutically acceptable carbohydrate, glucosamine, thiamine, niacinamide, an acidic buffer, a salt, and any combination thereof. 
     
     
         23 . The method according to  claim 22  wherein said acidic buffer is comprising acetic acid, benzoic acid, carboxylic acid, citric acid, hydrochloric acid, lactic acid, malic acid or tartaric acid. 
     
     
         24 . The method according to  claim 22  wherein said salt is calcium chloride, magnesium chloride, potassium chloride or sodium chloride. 
     
     
         25 . The method according to  claim 1  wherein the step of contacting the aqueous humor with the microplasmin comprises injecting the microplasmin into the vitreous or the aqueous humor. 
     
     
         26 . The method according to  claim 25  wherein the microplasmin is a human microplasmin. 
     
     
         27 . The method according to  claim 1  wherein the subject is a human. 
     
     
         28 . The method according to  claim 1  wherein the method is performed in the absence of mechanical vitrectomy. 
     
     
         29 . The method according to  claim 1  wherein the method is performed as an adjunct to mechanical vitrectomy. 
     
     
         30 . The method according to  claim 1  wherein the effective amount of the microplasmin is in the range of 0.005 mg to 0.2 mg per eye. 
     
     
         31 . The method according to  claim 1  wherein the microplasmin consists of a double chain polypeptide of amino acids 543 to 791 of SEQ ID NO:10 wherein the peptide bond between Arg561 and Val 562 is cleaved by a plasminogen activator. 
     
     
         32 . The method according to  claim 1  wherein the microplasmin is an activated microplasminogen wherein said microplasminogen is encoded by SEQ ID NO:3. 
     
     
         33 . The method according to  claim 1  wherein the microplasmin is produced by recombinant expression in a yeast. 
     
     
         34 . The method according to  claim 33  wherein said yeast is  Pichia pastoris.