Methods of preventing aggregation of various substances upon rehydration or thawing and compositions obtained thereby

Abstract

The present invention encompasses methods to prevent aggregation of a wide variety of substances during freezing/thawing and/or dehydrating/rehydrating. The substances thus obtained and compositions comprising the substances are also encompassed by the invention.

1 . A method of reducing aggregation during dehydration and rehydration of substances comprising the steps of 
 adding to a solution or suspension of the substances an amount of trehalose sufficient to prevent aggregation upon rehydration; and    dehydrating the solution or suspension.    
     
     
         2 . The method according to  claim 1  wherein the substances are selected from the group consisting of therapeutic, prophylactic and diagnostic.  
     
     
         3 . The method according to  claim 2  wherein the substances are therapeutic and are biological modifiers.  
     
     
         4 . The method according to  claim 3  wherein the biological modifier is selected from the group consisting of proteins and peptides, steroid hormones, oligosaccharides, nucleic acids and small molecules.  
     
     
         5 . The method according to  claim 4  wherein the proteins are selected from the group consisting of growth hormones, growth factors, insulin, monoclonal antibodies, interleukins and interferons.  
     
     
         6 . The method according to  claim 5  wherein the substance is human growth hormone.  
     
     
         7 . The method according to  claim 4  wherein the steroid hormones are selected from the group consisting of estrogen, progesterone and testosterone.  
     
     
         8 . The method according to  claim 2  wherein the substances are prophylactic substances and are aluminum based adjuvants.  
     
     
         9 . The method according to  claim 8  further comprising the step of incorporating the adjuvants into vaccines.  
     
     
         10 . The method according to  claim 9  wherein the vaccines are diphtheria/tetanus/pertussis (DTP) or inactivated poliovaccine.  
     
     
         11 . The method according to  claim 10  wherein the vaccine is DTP.  
     
     
         12 . The method according to  claim 2  wherein the substance is diagnostic and is selected from the group consisting of colloidal gold, polystyrene latex, fixed erythrocytes and monoclonal antibodies.  
     
     
         13 . The method according to  claim 12  wherein the substance is red blood cells, further comprising the step of fixing the red blood cells prior to adding trehalose.  
     
     
         14 . The method according to  claim 13  wherein the fixing is by glutaraldehyde.  
     
     
         15 . The method according to  claim 1  wherein the trehalose is added in an amount to obtain a final concentration of from about 1% to 50% (w/v).  
     
     
         16 . The method according to  claim 1  wherein the trehalose is added in an amount to obtain a final concentration of from about 5% to 25% (w/v).  
     
     
         17 . The method according to  claim 1  wherein the dehydration step occurs by lyophilization, drying at ambient conditions or drying under reduced vapor pressure.  
     
     
         18 . The method according to  claim 17  wherein the drying at reduced vapor pressure occurs at room temperature or at a temperature elevated above room temperature but below a temperature at which degradation or chemical change of the substance occurs.  
     
     
         19 . The method according to  claim 1  further comprising the step of rehydrating the substance to obtain a solution or suspension of substantially nonaggregated substance.  
     
     
         20 . A method of reducing aggregation of substances in solution or suspension during freezing comprising the steps of: 
 adding to the solution or suspension of the substance an amount of trehalose sufficient to prevent aggregation during freezing; and    freezing the solution or suspension.    
     
     
         21 . The method according to  claim 18  wherein the substances are selected from the group consisting of therapeutic, prophylactic and diagnostic.  
     
     
         22 . The method according to  claim 21  wherein the substance is therapeutic and is a biological response modifier.  
     
     
         23 . The method according to  claim 22  wherein the biological modifier is selected from the group consisting of proteins and peptides, steroid hormones, oligosaccharides, nucleic acids and small molecules.  
     
     
         24 . The method according to  claim 23  wherein the proteins are selected from the group consisting of growth hormones, growth factors, insulin, monoclonal antibodies, interleukins and interferons.  
     
     
         25 . The method according to  claim 24  wherein the substance is human growth hormone.  
     
     
         26 . The method according to  claim 23  wherein the steroid hormones are selected from the group consisting of estrogen, progesterone and testosterone.  
     
     
         27 . The method according to  claim 21  wherein the substances are prophylactic substances and are aluminum based adjuvants.  
     
     
         28 . The method according to  claim 27  further comprising the step of incorporating the adjuvants into vaccines.  
     
     
         29 . The method according to  claim 28  wherein the vaccines are diphtheria/tetanus/pertussis (DTP) or diphtheria/tetanus/pertussis/inactivated poliovaccine (DTP/IPV).  
     
     
         30 . The method according to  claim 29  wherein the vaccine is DTP/IPV.  
     
     
         31 . The method according to  claim 20  wherein the substance is diagnostic and is selected from the group consisting of colloidal gold, polystyrene latex, fixed erythrocytes and monoclonal antibodies.  
     
     
         32 . The method according to  claim 20  wherein the trehalose is added in an amount to attain of from about 1% to 50% (w/v).  
     
     
         33 . The method according to  claim 32  wherein the trehalose is added in an amount to attain of from about 5% to 25% (w/v).  
     
     
         34 . The method according to  claim 20  further comprising the step of thawing the solution or suspension to obtain a solution or suspension of substantially nonaggregated substance.  
     
     
         35 . An aqueous composition comprising a substance and an amount of trehalose sufficient to prevent substantial aggregation of the substance upon freezing and thawing or dehydrating and rehydrating.  
     
     
         36 . A frozen composition comprising a substance and an amount of trehalose sufficient to prevent substantial aggregation of the substance upon thawing.  
     
     
         37 . A dehydrated composition comprising a substance and an amount of trehalose sufficient to prevent aggregation of the substance upon rehydration.  
     
     
         38 . A composition obtained by the method according to  claim 1 .  
     
     
         39 . A composition obtained by the method according to  claim 13 .  
     
     
         40 . A composition obtained by the method according to  claim 19 .  
     
     
         41 . A composition obtained by the method according to  claim 20 .  
     
     
         42 . A composition obtained by the method according to  claim 34.