Mutated structural protein of a parvovirus

Abstract

The present invention is related to a structural protein of a parvovirus with an amino acid insertion at the insertion site I-453, a library comprising the protein, a multimeric structure comprising the protein, a nucleic acid encoding the protein, a vector, virus or cell comprising the nucleic acid, a process for the preparation of the protein, a medicament comprising the protein, nucleic acid or multimeric structure as well as methods and uses involving the protein, nucleic acid or multimeric structure.

1 - 51 . (canceled) 
     
     
         52 . A structural protein of a parvovirus which comprises an amino acid insertion of one or more amino acids into I-453. 
     
     
         53 . The structural protein of  claim 52 , wherein the amino acid insertion is directly C-terminally of amino acid G 453  in the sequence of AAV-2 or the corresponding amino acid of any other parvovirus. 
     
     
         54 . The structural protein of  claim 52 , wherein the insertion is located on the surface of the capsid formed by the structural proteins. 
     
     
         55 . The structural protein of  claim 52 , wherein the structural protein with the insertion is capable of particle formation. 
     
     
         56 . The structural protein of  claim 52 , wherein the parvovirus is an adeno-associated virus. 
     
     
         57 . The structural protein of  claim 56 , wherein the parvovirus is selected from the group consisting of AAV-1, AAV-2, AAV-3b, AAV-4, AAV-5, AAV-6, AAV-7, AAV-8, AAV-9, AAV-10, AAV-11, AAV-12, b-AAV, feline panleukopenia virus (FPV), canine parvovirus (CPV), B19, goose parvovirus (GPV) and minute virus of mice (MVM), especially FPV, CPV and B19. 
     
     
         58 . The structural protein of  claim 52 , wherein the amino acid insertion has a length of about 4 to about 30 amino acids. 
     
     
         59 . The structural protein of  claim 52 , wherein the amino acid insertion is selected from the group consisting of an epitope, a B-cell epitope and a tolerogen-derived epitope. 
     
     
         60 . The structural protein of  claim 52 , wherein the insertion is a part of a protein selected from the group consisting of a tumor antigen, a misfolded protein, a serum protein, a membrane protein, a viral receptor, a TNF-family member and an interleukin. 
     
     
         61 . The structural protein of  claim 59 , wherein the tolerogen-derived epitope is derived from a protein from the group consisting of CETP, CD20, acetylcholine receptors, IL13R, EGFR, IgE, Melan A, HMW MAA, CA125, Her2/NEU, CCR5, L1 cell adhesion molecule, VEGF, EGFR, CD20, TNF-α, IL-6, IL9, IL-13, IL-17, and β-amyloid. 
     
     
         62 . The structural protein of  claim 61 , wherein the tolerogen-derived epitope is selected from the group consisting of VNLTWSRASG, EFCINHRGYWVCGD, EDGQVMDVDLS, EKQRNGTLT, TYQCRVTHPHLPRALMR, RHSTTQPRKTKGSG, DSNPRGVSAYLSR, TITCLVVDLAPSK, KTKGSGFFVF, THPHLPRALMRS, GETYQCRVTHPHLPRALMRSTTK, LPRALMRS, INHRGYWV, CDAGSVRTNAPD AKAVSNLTESRSESLQS, SLTGDEFKKVLET, REAVAYRFEED, INPEIITLDG, DISVTGAPVITATYL, DISVTGAPVITA, PKTVSNLTESSSESVQS, SLMGDEFKAVLET, QHSVAYTFEED, INPEIITRDG, DISLTGDPVITASYL, DISLTGDPVITA, DQSIDFEIDSA, KNVSEDLPLPTFSPTLLGDS, KNVSEDLPLPT, CDSGRVRTDAPD, FPEHLLVDFLQSLS, DAEFRHDSG, HYAAAQWDFGNTMCQL, YAAQWDFGNTMCQ, RSQKEGLHYT, SSRTPSDKPVAHVVANPQAE, SRTPSDKPVAHVVANP, SSRTPSDKP, NADGNVDYHMNSVP, DGNVDYHMNSV, RSFKEFLQSSLRALRQ; FKEFLQSSLRA, and QMWAPQWGPD. 
     
     
         63 . The structural protein of  claim 52 , wherein the amino acid insertion brings about an increase in the transducing activity of the parvovirus. 
     
     
         64 . The structural protein of  claim 63 , wherein the amino acid insertion mediates binding of the structural protein to a cell membrane receptor. 
     
     
         65 . The structural protein of  claim 52 , wherein the amino acid insertion brings about an alteration in a chromatographic property of the structural protein. 
     
     
         66 . The structural protein of  claim 52 , wherein the parvovirus structural protein comprises one or more further mutation(s) at a site different to I-453 independently selected from a point mutation, an internal deletion, an N-terminal deletion, an insertion and a substitution. 
     
     
         67 . The structural protein of  claim 66 , wherein the further mutation is an insertion of a B-cell epitope or a cytotoxic T-cell epitope. 
     
     
         68 . The structural protein of  claim 66 , wherein the further mutation reduces the transducing activity of the particle for a given target cell by at least 50%. 
     
     
         69 . The structural protein of  claim 68 , wherein the further mutation is a mutation inactivating the HSPG binding site located in the proximity of I-587. 
     
     
         70 . The structural protein of  claim 66 , wherein the further mutation reduces the ability to induce a B-cell response against a parvovirus-specific epitope and/or mimotope. 
     
     
         71 . A nucleic acid encoding a structural protein of  claim 52 . 
     
     
         72 . A vector comprising a nucleic acid of  claim 71 . 
     
     
         73 . A method for altering the tropism of a parvovirus, the method comprising the steps of:
 a) co-expressing parvoviral helper and vector functions, wherein the helper function expresses a parvoviral structural protein according to  claim 52  under conditions that enable parvovirus formation, and   b) isolating the parvovirus.   
     
     
         74 . A method for vaccinating a mammal, the method comprising the vaccination of a mammal, with a structural protein according to  claim 52 . 
     
     
         75 . A method for breaking immune tolerance or treating an infectious disease, the method comprising the vaccination of a mammal with a structural protein according to  claim 52 . 
     
     
         76 . Medicament comprising at least one parvovirus structural protein according to  claim 52  or a nucleic acid according to  claim 71 . 
     
     
         77 . Method of treating and/or preventing a disease, the method comprising the step of administering to a subject need thereof a therapeutically effective amount of the medicament of  claim 76 , wherein the disease is selected from the group consisting of
 an allergic disease or asthma whereas at insertion site I-453 the structural protein of a parvovirus comprises an anti-idiotypic epi-/mimotope of an anti-IgE antibody or an IgE epi-/mimotope;   Alzheimer's disease whereas at insertion site I-453 the structural protein of a parvovirus comprises a β-amyloid epitope or mimotope;   atherosclerosis whereas at insertion site I-453 the structural protein of a parvovirus comprises a CETP epitope or mimotope;   a tumor disease whereas at insertion site I-453 the structural protein of a parvovirus comprises a tumor antigen;   an autoimmune disease or a chronic inflammatory disease, whereas at insertion site I-453 the structural protein of a parvovirus comprises an epitope or mimotope of a cytokine; and   an infectious disease whereas at insertion site I-453 the structural protein of a parvovirus comprises an epitope or mimotope of a viral receptor.