Anti-RAS intracellular binding proteins and use thereof

Details Anti-RAS intracellular binding proteins and use thereof Anti-RAS intracellular binding proteins and use thereof

1995-12-28

2000-12-12

Chambers, Jasemine

Drug, bio-affecting and body treating compositions

Designated organic active ingredient containing

Carbohydrate doai

4353201, 435325, 435455, 435 691, 435 7, 424 9321, 536 231, 536 234, 536 235, 536 2353, A01N 4304, A61K 3170, C12N 1563, C12N 1500

Patent

active

061599476

DESCRIPTION:

BRIEF SUMMARY
The present invention relates to nucleic acid sequences, to vectors containing them and to their therapeutic uses, in particular in gene therapy. More especially, the present invention relates to nucleic acid sequences comprising a gene coding for an intra-cellular binding protein (IBP) and to their use in gene therapy, where appropriate incorporated in suitable expression vectors.
Gene therapy consists in correcting a deficiency or abnormality (mutation, aberrant expression, and the like) by introduction of genetic information into the affected cell or organ. This genetic information may be introduced either in vitro into a cell extracted from the organ, the modified cell then being reintroduced into the body, or directly in vivo into the appropriate tissue. In this connection, different techniques of transfection and of gene transfer have been described in the literature (see Roemer and Friedman, Eur. J. Biochem. 208 (1992) 211). To date, the approaches proposed in the prior art for gene therapy consist in transferring genes coding for active polypeptides involved in genetic disorders (hormones, growth factors, and the like), antisense genes, or antigenic peptides for the production of vaccines. The present invention relates to a new approach to gene therapy, consisting in transferring and expressing in a target cell (or tissue) an intracellular peptide capable of interacting with cell components and thus of interfering with cell functions. The present invention is based more especially on the demonstration that it is possible to express in vivo modified antibodies which remain in the intracellular compartment and which can control certain cell functions. The invention is also based on the demonstration that it is possible to clone DNA sequences coding for such intracellular antibodies into vectors, in particular viral vectors, for use in gene therapy.
The use of antibodies in human therapy makes it possible, in general, to target and neutralize circulating biological complexes and/or those which are localized at the cell surface, by bringing about a cascade of events conducted by the immune system which leads to their removal. However, in many cases, including cancers or diseases due, for example, to viruses, this approach is fruitless since the antigen responsible for the deregulation of the affected cells is inaccessible to the injected antibodies. The present invention affords a new, especially advantageous therapeutic approach, consisting in causing antibodies or therapeutic agents whose binding to their epitope decreases and/or abolishes the deregulation to be produced continuously and intracellularly.
The possibility of recombinant expression of antibodies has already been described in the literature. Thus, Patent EP 88,994 describes the in vitro expression and purification of heavy or light-chain variable regions of antibodies. Likewise, U.S. Pat. No. 4,946,778 describes the in vitro expression of DNA sequences coding for modified antibodies composed of heavy- and light-chain variable regions of an antibody linked via a linker. However, the antibodies described in this patent are inactive, and generally synthesized in the form of insoluble inclusion bodies. The antibodies must hence be purified and then subjected to chemical treatments (denaturation, renaturations, and the like) in order to recover an activity. The present invention demonstrates the possibility of using such DNA sequences for the expression of active intracellular antibodies directly in vivo. The present invention thus demonstrates the possibility of using such DNA sequences coding for intracellular antibodies, under the control of regions permitting their expression in mammalian cells, for gene therapy, in particular in man. This new approach hence makes it possible to target cell components which are not accessible by traditional vaccination methods. Furthermore, this approach does not involve the development of an immune response, but acts intracellularly.
A first subject of the invention hence lies in a nucleic acid sequence

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