Modified Latency Associated Protein Construct

Abstract

The present invention provides a fusion protein comprising a latency associated peptide (LAP), a pharmaceutically active agent and an amino acid sequence comprising a dimerisation domain, wherein the LAP and the pharmaceutically active agent are connected by an amino acid sequence comprising a proleolytic cleavage site. Also provided are nucleic acids enclosing such fusion proteins, process for their preparation, pharmaceutical compositions, kits and uses thereof in medicine.

1 - 30 . (canceled) 
     
     
         31 . A protein dimer comprising a pair of fusion proteins, each fusion protein comprising a latency associated peptide (LAP) which is the precursor domain of TGFβ-1, -2, -3, -4, or -5, a pharmaceutically active agent and an amino acid sequence comprising a dimerisation domain comprising an antibody fragment polypeptide, wherein the LAP and the pharmaceutically active agent are connected by an amino acid sequence comprising a proteolytic cleavage site, and the dimerisation domain is linked to the N-terminus of the LAP, and wherein the fusion proteins are associated at the dimerisation domain in each fusion protein and form a closed shell around the pharmaceutically active agent, wherein the pharmaceutically active agent is an antibody or antibody fragment. 
     
     
         32 . The protein dimer as claimed in  claim 31 , wherein the antibody fragment polypeptide is an Fc region polypeptide, an immunoglobulin hinge polypeptide, a CH3 domain polypeptide, a CH4 domain polypeptide, a CH1 domain polypeptide, or a CL domain polypeptide. 
     
     
         33 . The protein dimer as claimed in  claim 31 , wherein the antibody fragment polypeptide is an Fc region polypeptide. 
     
     
         34 . The protein dimer as claimed in  claim 33 , wherein the Fc region polypeptide is derived from an IgG or IgA antibody. 
     
     
         35 . The protein dimer as claimed in  claim 31 , wherein the dimerisation domain is linked to the latency associated peptide by a linker sequence. 
     
     
         36 . The protein dimer as claimed in  claim 31 , wherein the proteolytic cleavage site is a matrix metalloproteinase or an aggrecanase cleavage site. 
     
     
         37 . The protein dimer as claimed in  claim 31 , wherein the antibody fragment is an scFv, Fab, Fab′, F(ab′) 2 , Fv, dsFv diabody, or Fd fragment. 
     
     
         38 . The protein dimer as claimed in  claim 31 , wherein the antibody or antibody fragment thereof is an anti-TNF antibody, anti-interleukin antibody, anti-interferon antibody, anti-cytokine antibody, or fragment thereof. 
     
     
         39 . The protein dimer as claimed in  claim 31 , wherein the antibody or antibody fragment thereof is an antibody against the HER2/neu receptor. 
     
     
         40 . The protein dimer as claimed in  claim 31 , wherein the antibody fragment is a trastuzumab scFv. 
     
     
         41 . A pharmaceutical composition comprising a protein dimer of  claim 31 . 
     
     
         42 . A nucleic acid construct encoding a fusion protein as defined in  claim 31  comprising a nucleic acid sequence encoding a pharmaceutically active agent, a nucleic acid sequence encoding a LAP, and a nucleic acid sequence encoding a dimerisation domain composed of an antibody fragment polypeptide, wherein the pharmaceutically active agent is an antibody or an antibody fragment. 
     
     
         43 . The nucleic acid construct as claimed in  claim 42 , wherein the dimerisation domain polypeptide is an Fc region polypeptide derived from an IgG or IgA antibody. 
     
     
         44 . The nucleic acid construct as claimed in  claim 42 , wherein the nucleic acid construct further comprises a nucleic acid sequence encoding a proteolytic cleavage site. 
     
     
         45 . The nucleic acid construct as claimed in  claim 42 , which is in the form of a vector. 
     
     
         46 . A fusion protein encoded by the nucleic acid construct of  claim 42 . 
     
     
         47 . A cell comprising a nucleic acid construct of  claim 42 . 
     
     
         48 . A pharmaceutical composition comprising a nucleic acid construct as claimed in  claim 42 . 
     
     
         49 . A method for the treatment of an inflammatory condition or cancer comprising the administration to a subject of a composition comprising a protein dimer of  claim 31 . 
     
     
         50 . A method for the treatment of an inflammatory condition or cancer comprising the administration to a subject of a composition comprising a nucleic acid construct of  claim 42 . 
     
     
         51 . A kit comprising a protein dimer as claimed in  claim 31  as an administration vehicle. 
     
     
         52 . A process for preparing the protein dimer as claimed in  claim 31 , comprising producing the fusion proteins recombinantly by expression in a host cell, purifying the expressed fusion proteins, and associating the dimerisation domain of the fusion proteins at the N-terminus of the LAP to form a closed shell around the pharmaceutically active agent. 
     
     
         53 . A process for preparing a nucleic acid construct of  claim 42 , comprising ligating together nucleic acid sequences encoding a latency associated peptide, a dimerisation domain, a proteolytic cleavage sequence, and a pharmaceutically active agent, optionally including a linker sequence on either side of the proteolytic cleavage site.