Camelidae antibodies against imminoglobulin e and use thereof for the treatment of allergic disorders

Abstract

The invention relates to a method suitable for administering protein therapeutic molecules orally, sublingually, topically, intravenously, subcutaneously, nasally, vaginally, rectally or by inhalation so as to avoid inactivation, by using VHH polypeptides derived from Camelidae antibodies. The invention further relates to the said therapeutic molecules. The invention further a method for delivering therapeutic molecules to the interior of cells. The invention furthe relates to anti-IgE therapeutic molecules.

1 . A method for delivering an anti-target compound to a subject for the treatment of a disorder without being inactivated by administering thereto a polypeptide construct comprising one or more single domain antibodies directed against said target.  
     
     
         2 . A method according to  claim 1  wherein said target is located in the gut system, and said polypeptide construct is delivered orally.  
     
     
         3 . A method according to  claim 1  wherein said target is located in vaginal and/or rectal tract, and said polypeptide construct is delivered to the vaginal and/or rectal tract.  
     
     
         4 . A method according to  claim 1  wherein said target is located in nose, upper respiratory tract and/or lung, and said polypeptide construct is delivered to nose, upper respiratory tract and/or lung.  
     
     
         5 . A method according to  claim 1  wherein said target is located in intestinal mucosa, and said polypeptide construct is delivered orally.  
     
     
         6 . A method according to  claim 1  wherein said target is located in the tissues beneath the tongue, and said polypeptide construct is delivered to the tissues beneath the tongue.  
     
     
         7 . A method according to  claim 1  wherein said target is located in the skin, and said polypeptide construct is delivered topically.  
     
     
         8 . A method according to  claim 1  wherein said target is in, or accessible via the blood, and said polypeptide construct is delivered orally, to the vaginal and/or rectal tract, nasally, by inhalation though the mouth or nose, to the tissues beneath the tongue, or topically.  
     
     
         9 . A polypeptide construct comprising at least one single domain antibody directed against a target, wherein the single domain antibody corresponds to a sequence represented by any of SEQ ID NOs: 12 to 70.  
     
     
         10 .- 14 . (canceled)  
     
     
         15 . A method according to  claim 1 , wherein said target is TNF-alpha and the disorder is inflammation.  
     
     
         16 . A method according to  claim 15 , wherein the single domain antibody corresponds to a sequence represented by any of SEQ ID NOs: 12 to 14.  
     
     
         17 . A method according to  claim 1 , wherein said target is CEA and the disorder is colon cancer.  
     
     
         18 . A method according to  claim 1 , wherein said target is EGFR and the disorder is any of head, neck, lung and colon cancer.  
     
     
         19 . A method according to  claim 18 , wherein the single domain antibody corresponds to a sequence represented by any of SEQ ID NOs: 23 to 44.  
     
     
         20 . A method according to  claim 1 , wherein said target is antigen of  Helicobacter pylori  and the disorder is any of indigestion and gastritis.  
     
     
         21 . A method according to  claim 1 , wherein said target is antigen of  Mycobacterium tuberculosis  and the disorder is tuberculosis.  
     
     
         22 . A method according to  claim 1 , wherein said target is antigen of influenza virus and the disorder is flu.  
     
     
         23 . A method according to  claim 1 , wherein said target is antigen of MMP and the disorder is cancer.  
     
     
         24 . A method according to  claim 23 , wherein the single domain antibody corresponds to a sequence represented by any of SEQ ID NOs: 15 to 22.  
     
     
         25 . A method according to  claim 1 , wherein said target is an antigen of IFN-gamma and the disorder is any of cancer, transplant rejection, auto immune disorder.  
     
     
         26 . A method according to  claim 25 , wherein the single domain antibody corresponds to a sequence represented by any of SEQ ID NOs: 45 to 70.  
     
     
         27 . A method according to  claim 1 , wherein said target is any of an antigen of  Helicobacter pylori , an antigen of  Mycobacterium tuberculosis , and an antigen of influenza virus.  
     
     
         28 . A polypeptide construct comprising (a) at least one single domain antibody directed against an internalising cellular receptor, and (b) at least one single domain antibody directed against a therapeutic target, or at least one therapeutic polypeptide or agent.  
     
     
         29 . (canceled)  
     
     
         30 . A polypeptide construct according to  claim 28  wherein said internalising cellular receptor is Epidermal Growth Factor receptor.  
     
     
         31 . A polypeptide construct according to  claim 30  wherein a single domain antibody directed against an internalising cellular receptor corresponds to a sequence represented by SEQ ID NO: 23 to 44.  
     
     
         32 . A polypeptide construct according to  claim 28  wherein said internalising cellular receptor is any of LDL receptor, FGF2 receptor, ErbB2 receptor, transferrin receptor, PDGF receptor, VEGF receptor, or PsmAr.  
     
     
         33 . A polypeptide construct according to  claim 28  wherein a single domain antibody directed against a therapeutic target, is directed against PDK1.  
     
     
         34 . (canceled)  
     
     
         35 . A polypeptide construct according to  claim 28  wherein a single domain antibody directed against a therapeutic target is directed against any of GSK1, Bad, caspase and Forkhead.  
     
     
         36 . (canceled)  
     
     
         37 . Method for delivering an anti-target therapeutic compound to the interior of a cell comprising administering to a subject a polypeptide construct according to  claim 28 .  
     
     
         38 . (canceled)  
     
     
         39 . A method according to  claim 37  wherein said cell is located in the gut system, and said polypeptide construct is delivered orally.  
     
     
         40 . A method according to  claim 37  wherein said cell is located in vaginal and/or rectal tract, and said polypeptide construct is delivered to the vaginal and/or rectal tract.  
     
     
         41 . A method according to  claim 37  wherein said cell is located in nose, upper respiratory tract and/or lung, and said polypeptide construct is delivered to nose, upper respiratory tract and/or lung.  
     
     
         42 . A method according to  claim 37  wherein said cell is located in intestinal mucosa, and said polypeptide construct is delivered orally.  
     
     
         43 . A method according to  claim 37  wherein said cell is located in the tissues beneath the tongue, and said polypeptide construct is delivered to the tissues beneath the tongue.  
     
     
         44 . A method according to  claim 37  wherein said cell is located in the skin, and said polypeptide construct is delivered topically.  
     
     
         45 . A method according to  claim 37  wherein said cell is in, or accessible via the blood, and said polypeptide construct is delivered orally, to the vaginal and/or rectal tract, nasally, by inhalation though the mouth or nose, to the tissues beneath the tongue, or topically.  
     
     
         46 . A polypeptide construct according to any of claims  9  or  28 , or as used in a method according to  claim 1 , wherein the single domain antibodies are humanized  Camelidae  VHHs an homologous sequence a functional portion or a functional portion of an homologous sequence of the full length single domain antibody or wherein the polypeptide construct is an homologous sequence a functional portion, or a functional portion of an homologous sequence of the full length polypeptide construct.  
     
     
         47 .- 49 . (canceled)  
     
     
         50 . A nucleic acid encoding a polypeptide construct according to any of claims  9 ,  28 , or  46 .  
     
     
         51 . A composition comprising a polypeptide construct according to any of claims  9 ,  28  or  46 , together with a pharmaceutical carrier.  
     
     
         52 . A method according to  claim 1 , wherein said target is antigen of IgE and the disorder is allergic response.  
     
     
         53 . A method according to  claim 52 , wherein a single domain antibody corresponds to a sequence represented by any of SEQ ID NOs: 1 to 11.  
     
     
         54 . A polypeptide construct comprising at least one single domain antibody directed against IgE.  
     
     
         55 . A polypeptide construct according to  claim 54  wherein at least one single domain antibody is a  Camelidae  VHH.  
     
     
         56 . A polypeptide construct according to  claim 54  wherein at least one single domain antibody corresponds to a sequence represented by any of SEQ ID NOs: 1 to 11.  
     
     
         57 . A polypeptide construct according to  claim 54 , wherein the number of anti-IgE single domain antibodies is at least two.  
     
     
         58 . A polypeptide construct according to  claim 57 , wherein at least one single domain antibody is a humanized  Camelidae  VHH.  
     
     
         59 . A polypeptide construct according to  claim 54 , wherein a single domain antibody is an homologous sequence, a functional portion, or a functional portion of an homologous sequence of the full length single domain antibody.  
     
     
         60 . A polypeptide construct according to  claim 54 , wherein the polypeptide construct is an homologous sequence, a functional portion, or a functional portion of an homologous sequence of the full length polypeptide construct.  
     
     
         61 . A nucleic acid encoding a polypeptide construct according to  claim 54 .  
     
     
         62 . A method for treating and/or preventing and/or alleviating disorders relating to inflammatory processes comprising administering to a subject in need of such treatment a polypeptide construct according to  claim 54 .  
     
     
         63 . A composition comprising a polypeptide construct according to  claim 54  together with a pharmaceutical carrier.