Vectors derived from antibodies for transferring substances into cells

Abstract

The present invention provides novel polypeptides which can effectively penetrate into cells thereby transporting a substance of interest into the cells.

1 . A polypeptide, characterized in that: 
 it is constituted by a unique or repeated peptide motif; and    it comprises an amino acid sequence endowing it with the capacity to penetrate into cells and, if necessary, to transport thereto a substance of interest.    
     
     
         2 . A polypeptide characterized in that: 
 it is constituted by a unique or repeated peptide motif;    it comprises an amino acid sequence constituted by one or more different antibody fragment(s); and    it is capable of penetrating into cells.    
     
     
         3 . A polypeptide according to  claim 1  or  claim 2 , characterized in that it comprises all or a portion of a hypervariable region of an antibody.  
     
     
         4 . A polypeptide according to  claim 1  or  claim 2 , characterized in that it comprises a fragment of a heavy antibody chain.  
     
     
         5 . A polypeptide according to  claim 4 , characterized in that it comprises all or a portion of the CDR3 region of an antibody.  
     
     
         6 . A polypeptide according to  claim 4  or  claim 5 , characterized in that it comprises all or a portion of the CDR2 region of an antibody.  
     
     
         7 . A polypeptide according to  claim 5  or  claim 6 , characterized in that it comprises all or a portion of the CDR3 region and all or a portion of the CDR2 region of an antibody.  
     
     
         8 . A polypeptide according to  claim 7 , characterized in that it essentially consists of a fusion between the CDR3 region of an antibody and the CDR2 region of an antibody.  
     
     
         9 . A polypeptide according to any one of the preceding claims, characterized in that it comprises at most 100 amino acids.  
     
     
         10 . A polypeptide according to  claim 9 , characterized in that it comprises 3 to 60 amino acids, preferably 3 to 30 amino acids.  
     
     
         11 . A polypeptide according to any one of the preceding claims, characterized in that the antibody fragment is a fragment of an antibody capable of penetrating into cells.  
     
     
         12 . A polypeptide according to  claim 11 , characterized in that the antibody fragment is a fragment of a polyreactive antibody.  
     
     
         13 . A polypeptide according to  claim 12 , characterized in that the antibody fragment is a fragment of an anti-DNA antibody.  
     
     
         14 . A polypeptide according to  claim 1  or  claim 2 , characterized in that it comprises a region with a sequence selected from SEQ ID No. 1, 2, 3 and 8, or any functional homologue.  
     
     
         15 . A polypeptide according to any one of the preceding claims, characterized in that it further comprises a region composed of basic amino acids, in particular lysine.  
     
     
         16 . A polypeptide according to  claim 1 , characterized in that the amino acid sequence is capable of being obtained by screening a peptide library for cell penetration.  
     
     
         17 . A polypeptide characterized in that it comprises a polylysine region and a region derived from a penetrating polyreactive antibody, and in that it is capable of penetrating into cells.  
     
     
         18 . A polypeptide according to any one of  claims 1  to  17 , with the capacity of reacting in vitro with anionic macromolecules such as double or single strand RNA, DNA, or with cationic macromolecules such as histones.  
     
     
         19 . A polypeptides according to any one of  claims 1  to  17 , with the capacity of reacting in vitro with heparin and heparin sulphate.  
     
     
         20 . A polypeptide according to  claim 1  or  claim 2 , characterized in that it is also capable of causing a substance to penetrate into a cell.  
     
     
         21 . Use of a polypeptide according to any one of the preceding claims, for preparing a composition intended to transfer substances into cells.  
     
     
         22 . Use of a polypeptide according to any one of  claims 1  to  20  for the preparation of an antiviral composition.  
     
     
         23 . A polypeptide according to any one of  claims 1  to  20 , characterized in that it is coupled to a substance.  
     
     
         24 . A vector for transferring a substance into a cell, characterized in that it comprises a polypeptide according to any one of  claims 1  to  20  to which said substance is coupled.  
     
     
         25 . A vector according to  claim 24 , characterized in that the coupling is a covalent coupling.  
     
     
         26 . A vector according to  claim 25 , characterized in that coupling is effected by a covalent maleimide, succinimide, peptide, disulphide, amine, acid, biotin-streptavidin or p-benzoquinone covalent type bond.  
     
     
         27 . A vector according to  claim 24 , characterized in that said substance is a nucleic acid.  
     
     
         28 . A vector according to  claim 24 , characterized in that said substance is a protein.  
     
     
         29 . A vector according to  claim 24 , characterized in that said substance is a drug.  
     
     
         30 . A vector according to  claim 24 , characterized in that said substance is an antigen.  
     
     
         31 . A eukaryotic cell containing a polypeptide according to any one of  claims 1  to  20 .  
     
     
         32 . A eukaryotic cell containing a vector according to  claim 21 .  
     
     
         33 . A method for transferring a substance into a cell in vitro, ex vivo or in vivo comprising: 
 coupling said substance to a polypeptide as defined in  claim 1 ,  2  or  17 ; and    incubating the cell with the product of said coupling.    
     
     
         34 . A method according to  claim 33 , characterized in that the cell is incubated with the coupling product in the presence of glycerol.  
     
     
         35 . A pharmaceutical composition comprising a vector according to  claim 24  in which the substance is an active principle of a drug, in association with a physiologically acceptable vehicle.  
     
     
         36 . A vaccine comprising a vector according to  claim 24  in which the substance is an antigen, in association with a physiologically acceptable vehicle.  
     
     
         37 . An antiviral composition comprising a polypeptide according to any one of  claims 1  to  20  or an antibody or an antibody fragment according to  claim 13 .  
     
     
         38 . Use of an antibody or antibody fragment according to  claim 13  for the preparation of an antiviral composition.  
     
     
         39 . Use according to  claim 38 , characterized in that the antibodies are monoclonal antibodies.  
     
     
         40 . Use according to  claim 22  or  claim 38 , in combination with an antiviral agent.  
     
     
         41 . Use according to  claim 22  or  claim 38 , characterized in that the virus is the human acquired immunodeficiency virus (HIV), a polio virus, a herpes virus or a cytomegalovirus.  
     
     
         42 . A method for modifying a cell with the aim of improving the resistance of that cell to a virus, comprising bringing said cell into contact with one or more polypeptides according to any one of  claims 1  to  20 , or polyreactive antibodies or antibody fragments having the capacity to bind DNA.  
     
     
         43 . A composition comprising cells incubated ex vivo in the presence of one or more polypeptides according to any one of  claims 1  to  20  or antibodies or antibody fragments as defined in  claim 13 .  
     
     
         44 . A composition according to  claim 43 , characterized in that the cells are human peripheral blood mononuclear cells.