Lyophilized formulation of tat-nr2b9c with acetylation scavenger

Abstract

The present invention provides lyophilized formulations of active agents, particularly of TAT-NR2B9c including histidine, trehalose and lysine. TAT-NR2B9c has shown promise for treating stroke, aneurysm, subarachnoid hemorrhage and other neurological or neurotraumatic conditions. Such formulations are stable at ambient temperature thus facilitating maintenance of supplies of such a formulation in ambulances for administration at the scene of illness or accident or in transit to a hospital.

1 . A prelyophilized formulation comprising a peptide, which has the amino acid sequence of SEQ ID NO:6 or SEQ ID NO:7 or differs therefrom by up to 5 substitutions, deletions or insertions, histidine, trehalose and an acetylation scavenger at a pH of 6-7. 
     
     
         2 . The prelyophilized formulation of  claim 1 , wherein the peptide is TAT-NR2B9c (SEQ ID NO:6). 
     
     
         3 . The prelyophilized formulation of  claim 1 , wherein the acetylation scavenger is lysine. 
     
     
         4 . The prelyophilized formulation of  claim 3 , wherein the TAT-NR2B9c is at a concentration of 70-120 mg/ml, the histidine is at a concentration of 15-100 mM, and the trehalose is at a concentration of 80-160 mM and the lysine is at a concentration of 300-900 mM. 
     
     
         5 . The prelyophilized formulation of  claim 1 , wherein the TAT-NR2B9c is at a concentration of 70-120 mg/ml, the histidine is at a concentration of 20-100 mM, the trehalose is at a concentration of 100-140 mM and the lysine is at a concentration of 500-800 mM. 
     
     
         6 . The prelyophilized formulation of  claim 5 , wherein the Tat-NR2B9c is at a concentration of 70-120 mg/ml, the concentration of histidine 20-50 mM, the concentration of trehalose is 100-140 mM and the concentration of lysine is 700-750 mM. 
     
     
         7 . The prelyophilized formulation of  claim 6 , wherein the concentration of histidine is 20 mM and the concentration of trehalose is 100-200 mM, preferably 120 mM, the concentration of TAT-NR2B9c is 90 mg/ml and the concentration of lysine is 720 mM. 
     
     
         8 . A lyophilized formulation prepared by lyophilizing the prelyophilized formulation of  claim 1 . 
     
     
         9 . A reconstituted formulation prepared by combining the lyophilized formulation of  claim 8  with an aqueous solution. 
     
     
         10 . The reconstituted formulation of  claim 9 , wherein the aqueous solution is water or normal saline. 
     
     
         11 . The reconstituted formulation of  claim 10 , wherein the volume of the reconstituted formulation is 3-6 times the volume of the prelyophilized formulation. 
     
     
         12 . A reconstituted formulation comprising TAT-NR2B9c at concentration of 15-25 mg/ml, histidine at a concentration of 4-20 mM and trehalose at a concentration of 20-30 mM and lysine at a concentration of 50-900 mM at pH 6-7. 
     
     
         13 . A method of preparing a formulation, comprising
 storing a lyophilized formulation sample according to  claim 4  for at least a week at ambient temperature; and   reconstituting the lyophilized formulation.   
     
     
         14 . The method of  claim 13 , wherein the storage includes periods in which the temperature exceeds 37° C. 
     
     
         15 . The method of  claim 13 , further comprising administering the reconstituted formulation, optionally after further dilution in normal saline or water, to a patient. 
     
     
         16 . The method of  claim 13 , wherein the patient has stroke or traumatic injury to the CNS. 
     
     
         17 . The method of  claim 16 , wherein the lyophilized sample is stored in an ambulance. 
     
     
         18 . The method of  claim 13 , wherein the patient has a subarachnoid hemorrhage. 
     
     
         19 . The method of  claim 13 , wherein the patient is undergoing endovascular repair for an aneurysm.