Stable formulations for cns delivery of arylsulfatase a

Abstract

The present invention provides, among oilier things, compositions and methods for CNS delivery of lysosomal enzymes (e.g., recombinant human arylsulfatase A (rhASA)) for effective treatment of lysosomal storage diseases (e.g. Metachromatic Leukodystrophy Disease). In some embodiments, the present invention includes a stable formulation for intrathecal administration comprising an ASA protein and a poloxamer, wherein less than 3% of the ASA protein exists in aggregated form.

1 - 47 . (canceled) 
     
     
         48 . A stable formulation for intrathecal administration comprising an arylsulfatase A (ASA) protein and a poloxamer, wherein the ASA protein is present at a concentration of at least approximately 0.1 mg/ml, and wherein less than 5% of the arylsulfatase A (ASA) protein exists in aggregated form. 
     
     
         49 . The stable formulation of  claim 48 , wherein the poloxamer is selected from the group consisting of Poloxamers 101, 105, 108, 122, 123, 124, 181, 182, 183, 184, 185, 188, 212, 215, 217, 231, 234, 235, 237, 238, 282, 284, 288, 331, 333, 334, 335, 338, 401, 402, 403, 407, and combination thereof. 
     
     
         50 . The stable formulation of  claim 49 , wherein the poloxamer is poloxamer 188. 
     
     
         51 . The stable formulation of  claim 48 , wherein the poloxamer is present at a concentration ranging from approximately 0.05-0.50%, ranging from approximately 0.05-0.20%, of approximately 0.15%, or of approximately 0.1%. 
     
     
         52 . The stable formulation of  claim 48 , wherein less than 4%, less than 3%, less than 2%, or less than 1% of the arylsulfatase A (ASA) protein exists in aggregated form. 
     
     
         53 . The stable formulation of  claim 48 , wherein the ASA protein is present at a concentration selected from about 1 mg/ml, 10 mg/ml, 30 mg/ml, 50 mg/ml, or 100 mg/ml. 
     
     
         54 . The stable formulation of  claim 48 , wherein the ASA protein comprises an amino acid sequence having at least 80% identity to SEQ ID NO:1 or the ASA protein comprises the amino acid sequence of SEQ ID NO:1. 
     
     
         55 . The stable formulation of  claim 48 , wherein the ASA protein is produced from a human cell line or from CHO cells. 
     
     
         56 . The stable formulation of  claim 48 , wherein the formulation further comprises salt. 
     
     
         57 . The stable formulation of  claim 56 , wherein the salt is NaCl and is present at a concentration ranging from approximately 0-300 mM, ranging from approximately 80-160 mM, or of approximately 154 mM. 
     
     
         58 . The stable formulation of  claim 48 , wherein the stable formulation further comprises a buffering agent selected from the group consisting of phosphate, acetate, histidine, succinate, citrate, Tris, and combinations thereof. 
     
     
         59 . The stable formulation of  claim 58 , wherein the buffering agent is phosphate. 
     
     
         60 . The stable formulation of  claim 48 , wherein the formulation has a pH of approximately 3-8.0, approximately 6.0-6.5, or approximately 6.0. 
     
     
         61 . The stable formulation of  claim 48 , wherein the formulation is a liquid formulation or is formulated as lyophilized dry powder. 
     
     
         62 . The stable formulation of  claim 48 , wherein the formulation further comprises a stabilizing agent selected from the group consisting of sucrose, glucose, mannitol, sorbitol, polyethylene glycol (PEG), histidine, arginine, lysine, phospholipids, trehalose and combination thereof. 
     
     
         63 . A stable formulation for intrathecal administration comprising
 an arylsulfatase A (ASA) protein at a concentration ranging from approximately 0.1-100 mg/ml, a poloxamer at a concentration ranging from approximately 0.05-0.50%, NaCl at a concentration ranging from approximately 0-300 mM, and a pH of 3-8.0, wherein less than 5% of the ASA protein exists in aggregated form.   
     
     
         64 . A container comprising a single dosage form of a stable formulation according to  claim 48 . 
     
     
         65 . The container of  claim 64 , wherein the container is selected from an ampule, a vial, a cartridge, a reservoir, a lyo-ject, or a pre-filled syringe. 
     
     
         66 . The container of  claim 64 , wherein the stable formulation is present in a volume of less than about 50.0 ml or less than about 5.0 ml. 
     
     
         67 . A method of treating metachromatic leukodystrophy (MLD) disease comprising a step of
 administering intrathecally to a subject in need of treatment a formulation comprising an arylsulfatase A (ASA) protein at a concentration ranging from approximately 0.1-100 mg/ml, a poloxamer at a concentration ranging from approximately 0.05-0.50%, NaCl at a concentration ranging from approximately 0-300 mM, and a pH of 3-8.0, wherein less than 5% of the ASA protein exists in aggregated form.