Lipopolysaccharide binding protein derivatives

Abstract

Disclosed are novel biologically active lipopolysaccharide binding protein (LBP) derivatives including LBP derivative hybrid proteins which are characterized by the ability to bind to and neutralize LPS and which lack the CD14-mediated immunostimutlatory properties of holo-LBP.

What is claimed is:  
     
         1 . An LBP protein derivative having an ability to bind to LPS and lacking CD14-mediated immunostimulatory properties.  
     
     
         2 . The derivative according to  claim 1  which has a molecular weight of about 25 kD.  
     
     
         3 . The derivative according to  claim 1  consisting of a portion of the amino-terminal half of LBP.  
     
     
         4 . The derivative according to  claim 3  consisting of amino acids 1 through 197 of SEQ ID NO: 1.  
     
     
         5 . The derivative according to  claim 1  which is LBP (1-197) (Cys131).  
     
     
         6 . A pharmaceutical composition comprising an LBP protein derivative according  claim 1  and a pharmaceutically acceptable diluent, adjuvant or carrier.  
     
     
         7 . A DNA sequence encoding an LBP derivative according to  claim 1 .  
     
     
         8 . A DNA vector comprising the DNA sequence according to  claim 7 .  
     
     
         9 . A host cell stably transformed or transfected with a DNA sequence according to  claim 7  in a manner allowing expression in the host cell of the protein encoded thereby.  
     
     
         10 . A method of treating a gram-negative bacterial infection and the sequelae thereof comprising administering an LBP protein derivative according to  claim 1 .  
     
     
         11 . The method of  claim 10  wherein the LBP protein derivative is administered at a dosage of from about 0.1 mg/kg to about 100 mg/kg of body weight.  
     
     
         12 . An LBP derivative hybrid protein having an ability to bind to LPS and lacking CD14-mediated immunostimulatory properties and characterized by the presence of at least a portion of an LPS binding domain of BPI selected from the group consisting of: 
 ASQQGTAALQKELKRIKPDYSDSFKIKH (SEQ ID NO: 17);    SSQISMVPNVGLKFSISNANIKISGKWKAQKRFLK (SEQ ID NO: 18); and    VHVHISKSKVGWLIQLFHKKIESALRNK (SEQ ID NO: 19).    
     
     
         13 . The hybrid protein according to  claim 12  characterized by the presence of at least a portion of an LPS binding domain of LBP selected from the group consisting of: 
 AAQEGLLALQSELLRITLPDFTGDLRIPH (SEQ IS NO: 20);  
 HSALRPVPGQGLSLSISDSSIRVQGRWKVRKSFFK (SEQ ID NO: 21); and  
 VEVDMSGDLGWLLNLFHNQIESKFQKV (SEQ ID NO: 22).  
 
     
     
         14 . The hybrid protein according to  claim 12  consisting of a portion of the amino-terminal half of LBP/and a portion of the amino-terminal half of BPI.  
     
     
         15 . The hybrid protein according to  claim 12  which is LBP(1-43)/BPI(44-199).  
     
     
         16 . The hybrid protein according to  claim 12  which is BPI(1-159)/LBP(158-197).  
     
     
         17 . The hybrid protein according to  claim 12  which is LBP(1-43)/BPI(44-159)/LBP(158-197).  
     
     
         18 . The hybrid protein according to  claim 12  which is BPI(1-137)/LBP(137-197).  
     
     
         19 . The hybrid protein according to  claim 12  which is BPI(1-25)/LBP(26-135)/BPI(137-199).  
     
     
         20 . The hybrid protein according to  claim 12  which is [LBP(1-87)/BPI (88-100)/LBP(101-197)].  
     
     
         21 . The hybrid protein according to  claim 12  which is [LBP(1-146)/BPI (148-161)/LBP(160-197)].  
     
     
         22 . The hybrid protein according to  claim 12  which is [LBP(1-87)/BPI(88-100)/LBP(101-146)/BPI(148-161)/LBP(160-197)].  
     
     
         23 . The hybrid protein according to  claim 12  which is [BPI(1-85)/LBP(86-99)/BPI(100-199)].  
     
     
         24 . The hybrid protein according to  claim 12  which is [BPI(1-147/LBP(147-159)/BPI(162-199)].  
     
     
         25 . The hybrid protein according to  claim 12  which is [BPI(1-85)/LBP(86-99)/BPI(100-147)/LBP(147-159)/BPI(162-199)].  
     
     
         26 . An LBP derivative hybrid protein having an ability to bind LPS and lacking CD-14 mediated immunostimulatory properties which is an LBP/IgG fusion protein.  
     
     
         27 . A pharmaceutical composition comprising an LBP derivative hybrid protein according to  claim 12  or  26  and a pharmaceutically acceptable diluent, adjuvant or carrier.  
     
     
         28 . A DNA sequence encoding an LBP derivative hybrid protein according to  claim 12  or  26 .  
     
     
         29 . A DNA vector comprising the DNA sequence according to  claim 28 .  
     
     
         30 . A host cell stably transformed or transfected with a DNA sequence according to  claim 29  in a manner allowing expression in the host cell of the protein encoded thereby.  
     
     
         31 . A method of treating a gram-negative bacterial infection and the sequelae thereof comprising administering an LBP derivative hybrid protein of  claim 12  or  26 .  
     
     
         32 . The method of  claim 31  wherein the LBP derivative hybrid protein is administered at a dosage of from about 0.1 mg/kg to about 100 mg/kg of body weight.