Treatment of microbial infections

Abstract

The present invention is directed to improved microbial antigen vaccines, pharmaceutical compositions, immunogenic compositions and antibodies and their use in the treatment of microbial infections, particularly those of bacterial origin, including Staphylococcal origin. Ideally, the present invention is directed to a recombinant staphylococcal MSCRAMM or MSCRAMM-like proteins, or fragment thereof, with reduced binding to its host ligand, for use in therapy.

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         25 . A method of inducing an immune response in an individual, the method comprising administering to the individual a recombinant Staphylococcal fibrinogen binding protein, or fibrinogen binding-like protein, or fragment thereof, the protein or fragment thereof comprising at least part of the fibrinogen binding region, and having at least one nucleotide or amino acid residue substitution, insertion, deletion or addition in the fibrinogen binding region that results in a recombinant fibrinogen binding protein with reduced ability or lacking the ability to non-covalently bind fibrinogen. 
     
     
         26 . A method of treating a patient having a microbial infection caused by Staphylococci, the method comprising treating said patient with a recombinant Staphylococcal fibrinogen binding protein or fibrinogen binding-like protein, or fragment thereof, or vaccine, the protein or fragment thereof or vaccine comprising at least part of the fibrinogen binding region, and having at least one nucleotide or amino acid residue substitution, insertion, deletion or addition in the fibrinogen binding region that results in a recombinant fibrinogen binding protein with reduced ability or lacking the ability to non-covalently bind fibrinogen, such that said microbial infection is treated. 
     
     
         27 . A nucleic acid construct, expression vector, or host cell expressing a recombinant Staphylococcal fibrinogen binding protein or fibrinogen binding-like protein, or fragment thereof, the protein or fragment thereof comprising at least part of the fibrinogen binding region, and having at least one nucleotide or amino acid residue substitution, insertion, deletion or addition in the fibrinogen binding region that results in a recombinant fibrinogen binding protein with reduced ability, or lacking the ability, to non-covalently bind fibrinogen. 
     
     
         28 . A vaccine comprising a recombinant Staphylococcal fibrinogen binding protein or fibrinogen binding-like protein, or fragment thereof, the protein or fragment thereof comprising at least part of the fibrinogen binding region, and having at least one nucleotide or amino acid residue substitution, insertion, deletion or addition in the fibrinogen binding region that results in a recombinant fibrinogen binding protein with reduced ability, or lacking the ability, to non-covalently bind fibrinogen. 
     
     
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         30 . An immunogenic pharmaceutical composition comprising a recombinant Staphylococcal fibrinogen binding protein, or fibrinogen binding-like protein, or fragment thereof, the protein or fragment thereof comprising at least part of the fibrinogen binding region, and having at least one nucleotide or amino acid residue substitution, insertion, deletion or addition in the fibrinogen binding region that results in a recombinant fibrinogen binding protein with reduced ability, or lacking the ability, to non-covalently bind fibrinogen, and a pharmaceutically acceptable adjuvant. 
     
     
         31 . The immunogenic pharmaceutical composition of  claim 30  wherein the recombinant Staphylococcal fibrinogen binding protein is ClfA. 
     
     
         32 . The immunogenic pharmaceutical composition of  claim 30  wherein the recombinant Staphylococcal fibrinogen binding protein has the amino acid sequence according to any of SEQ ID Nos. 1 to 3, or a fragment thereof, wherein one or both of residues P 336  and Y 338  are each substituted with either serine or alanine, or comprises the amino acid sequence according to any of SEQ ID NOs. 4 to 14. 
     
     
         33 . The immunogenic pharmaceutical composition of  claim 30  wherein the recombinant Staphylococcal fibrinogen binding protein is rClfAP 336 S Y 338 A or rClfAP 336  A Y 338 S or a fragment thereof. 
     
     
         34 . The method of  claim 26 , wherein the non-covalent binding that takes place during dock, lock and latching of the recombinant Staphylococcal fibrinogen binding protein or fibrinogen binding-like protein to fibrinogen is reduced or prevented. 
     
     
         35 . The method of  claim 26  wherein the recombinant Staphylococcal fibrinogen binding protein is selected from the group consisting of:
 FbI, SdrF, SdrG, fibrinogen binding protein clumping factor A (ClfA), fibrinogen binding protein clumping factor B (ClfB), IsdA, FnBPA, and FnBPB. 
 
     
     
         36 . The method of  claim 35 , wherein the recombinant Staphylococcal fibrinogen binding protein comprises the fibrinogen binding region, the minimal fibrinogen binding region or a fragment thereof. 
     
     
         37 . The method of  claim 35  wherein the recombinant Staphylococcal fibrinogen binding protein comprises:
 subregions N123, spanning amino acid residues 40 to 559 of the fibrinogen binding region (Region A); 
 subregions N23, spanning amino acid residues 221 to 559 of the fibrinogen binding region of ClfA (Region A); or 
 amino acid residues 221 to 531 of the fibrinogen binding region (Region A). 
 
     
     
         38 . The method of  claim 35  wherein the recombinant Staphylococcal fibrinogen binding protein has the amino acid sequence according to any of SEQ ID NOs. 1 to 3 or a fragment thereof wherein one or both of residues P 336  and Y 338  are each substituted with either serine or alanine. 
     
     
         39 . The method of  claim 35  wherein the recombinant Staphylococcal fibrinogen binding protein is rClfAP 336 S Y 338 A or rClfAP 336  A Y 338 S or a fragment thereof. 
     
     
         40 . The method of  claim 26  wherein the recombinant Staphylococcal fibrinogen binding protein is ClfA. 
     
     
         41 . The method of  claim 26  wherein the recombinant Staphylococcal fibrinogen binding protein is ClfA and one or more of amino acid residues Ala254, Tyr256, Pro336, Tyr338,Ile387, Lys389, Glu526 and Val527 are each independently substituted with either Ala or Ser. 
     
     
         42 . The method of  claim 26  wherein the nucleotide or amino acid substitution, insertion, deletion, or addition reduces the non-covalent interaction with fibrinogen by preventing or reducing fibrinogen binding to the hydrophobic pocket separating Region A subregions N2 and N3 of the fibrinogen binding protein. 
     
     
         43 . The method of  claim 26  wherein the recombinant Staphylococcal recombinant fibrinogen binding protein comprises the fibrinogen binding region without the latching peptide amino acid residues. 
     
     
         44 . The method of  claim 26  wherein the recombinant fibrinogen binding protein comprises the amino acid sequence according to any of SEQ ID Nos. 4 to 14. 
     
     
         45 . The nucleic acid construct, expression vector or host cell of  claim 27  wherein the recombinant staphylococcal fibrinogen binding protein is ClfA. 
     
     
         46 . The nucleic acid construct, expression vector or host cell of  claim 27  wherein the recombinant Staphylococcal fibrinogen binding protein has the amino acid sequence according to any of SEQ ID NOs. 1 to 3, or a fragment thereof, wherein one or both of residues P 336  and Y 338  are each substituted with either serine or alanine, or comprises the amino acid sequence according to any of SEQ ID NOs. 4 to 14. 
     
     
         47 . The nucleic acid construct of  claim 27  wherein the recombinant Staphylococcal fibrinogen binding protein is rClfAP 336 S Y 338 A or rClfAP 336 A Y 338 S or a fragment thereof. 
     
     
         48 . The vaccine of  claim 28  wherein the recombinant staphylococcal fibrinogen binding protein is ClfA. 
     
     
         49 . The vaccine of  claim 28  wherein the recombinant Staphylococcal fibrinogen binding protein has the amino acid sequence according to any of SEQ ID Nos. 1 to 3, or a fragment thereof, wherein one or both of residues P 336  and Y 338  are each substituted with either serine or alanine. 
     
     
         50 . The vaccine of  claim 28  wherein the recombinant Staphylococcal fibrinogen binding protein is rClfAP 336 S Y 338 A or rClfAP 336 A Y 338 S or a fragment thereof.