Ob protein compositions and methods

Abstract

The present invention provides methods and compositions for, for example, effecting weight loss and treating excess weight by administering OB protein, or a derivative thereof, in a form for constant supply, for example, continuous administration, at a dosage of less than or equal to about 1 mg protein/kg body weight/day. Compositions and methods used for production of recombinant murine and human OB protein are also provided. Compositions and methods for preparing recombinant murine methionyl OB protein and recombinant human methionyl OB protein, including DNA sequences, vectors, host cells, methods of fermentation, and methods of purification are provided herein.

1 . A method of treating excess weight in a mammal by continuous administration of 1 mg protein/kg body weight/day or less of an OB protein selected from the group consisting of:
 (a) recombinant methionyl murine OB protein (SEQ. ID. No. 2);   (b) recombinant methionyl human OB protein (SEQ ID No. 1);   (c) the protein of (a) or (b) lacking the methionyl residue at position −1;   (d) the protein of (a), (b) or (c) lacking a glutamine at position 28; and   (e) a chemically modified derivative of (a), (b), (c) or (d).   
     
     
         2 . A method of  claim 1  wherein the chemically modified derivative is a pegylated derivative. 
     
     
         3 . A method of  claim 2  wherein the pegylated derivative is N-terminally pegylated. 
     
     
         4 . A method of  claim 1  wherein said continuous administration is accomplished by osmotic pump. 
     
     
         5 . A DNA sequence according to SEQ ID No. 1. 
     
     
         6 . A vector containing a DNA sequence according to  claim 5 . 
     
     
         7 . A vector of  claim 6  wherein said vector is pCFM1656. 
     
     
         8 . A DNA sequence according to SEQ ID No. 3. 
     
     
         9 . A vector containing a DNA sequence according to  claim 8 . 
     
     
         10 . A vector according to  claim 9  wherein said vector is pCFM1656. 
     
     
         11 . A method of refolding partially purified OB protein in a solution obtained from inclusion bodies, said partially purified OB protein selected from the group consisting of:
 (a) recombinant methionyl murine OB protein (SEQ. ID. No. 2);   (b) recombinant methionyl human OB protein (SEQ ID No. 1);   (c) the protein of (a) or (b) lacking the methionyl residue at position −1;   wherein said refolding is accomplished using N-lauroyl sarcosine.   
     
     
         12 . A method of  claim 11  wherein said sarcosine is used at a concentration of 0.5%-2.0% weight per volume of solution.