Patent EA-001609-B1 (EA)
Title: Vehicle for the transport of substances into eukaryotic cells and process for the preparation the same
Inventor: BERTLING WOLR
IPC: A61K47/48
Filed: 1997-05-07
Abstract: 1. A process for the preparation of a vehicle for the transport into the eukaryotic cells, such as DNA, RNA, protein, PNA or or pharmaceuticals of lipophilic and lipophobic character, wherein said vehicle comprising at least one capsomere which is formed such that it is suitable for the construction of a capsid or a capsid-like structure and  on its inner side has a structure interacting with the molecular substance such that the molecular substance can be bound or added to the capsomere, comprising the following steps: i) synthesis, purification or isolation of the capsomere and ii) complexation of the molecular substance using the capsomere. 2. The process as claimed in claim 1, where the capsomere is a capsomere of a polyoma virus or derived therefrom. 3. The process as claimed in claim 2, where the capsomere is formed from the VP1 pentamer of the polyoma virus or is derived therefrom. 4. The process as claimed in claim 1, where the capsomere is obtained from "non-enveloped" viruses such as DNA-containing Papovaviridae, in particular polyoma and papilloma viruses, Iridoviridae, Adenoviridae, Parvoviridae or RNA-containing Picornaviridae, in particular polio viruses, Caliciviridae, Reoviridae and Birnaviridae or is derived therefrom. 5. The process as claimed in claim 1, where the capsomere is obtained from the outer and/or inner coat of "enveloped" viruses such as DNA-containing Poxviridae, Herpesviridae, Hepadnaviridae or RNA-containing Retroviridae, Paramyxoviridae, Sendai viruses, Orthomyxoviridae, Bunyaviridae, Arenaviridae, Toroviridae, Togaviridae, Flaviviridae, Rhabdoviridae and Filoviridae or is derived therefrom. 6. The process as claimed in one of the preceding claims, where the interactions of the substance and the capsomere are lipophilic interactions and/or are based on covalent bonds, ionic bonds or hydrogen bridges. 7. The process as claimed in one of the preceding claims, where the structure interacting with the substance comprises bifunctional, preferably heterologous bifunctional, groups. 8. The process as claimed in claim 7, where the bifunctional groups are selected from the substance group consisting of maleimide derivatives, alkyl halides, aryl halides, isocyanates, glutaraldehydes, acrylating reagents and imidoesters. 9. The process as claimed in claim 7 or 8, where the bifunctional groups react with cysteine residues on the capsomere. 10. The process as claimed in one of the preceding claims, where the structure interacting with the substance comprises affinity-increasing groups such as 4-iodoacetamidosalicylic acid and/or p-arsonic acid phenyldiazonium fluroborate and/or derivatives thereof. 11. The process as claimed in one of claims 3 to 10, where the structure interacting with the substance is formed by epitopes of the VP1 pentamer. 12. The process as claimed in one of the preceding claims, where, using at least one further capsomere, it is possible to prepare a capsid-like structure for the transport of the molecular substance into a prespecified type of cells or to a prespecified site in the intracellular region. 13. The process as claimed in one of the preceding claims, where the capsid-like structure comprises at least one VP-2 and/or VP-3 protein. 14. The process as claimed in one of the preceding claims, where after step lit.i [sic] the suitable residues of the capsomere, in particular its cysteine residues, are modified with bifunctional groups. 15. The process as claimed in one of the preceding claims, where the modification is carried out using one or more of the following substances: maleimide derivatives, alkyl halides, aryl halides, isocyanates, glutaraldehydes, acrylating reagents and imidoesters. 16. The use of the vehicle prepared by the process as claimed in one of the preceding claims as a pharmaceutical carrier for the administration of molecules such as DNA, RNA, oligonucleotides, PNA, proteins, peptides and also lower molecular weight lipophilic and lipophobic reagents, colloidal gold and gold-labeled proteins and peptides to eukaryotic cells.