Patent EA-004062-B1 (EA)
Title: Modulators of the function of receptors of the tnf/ngf receptor family and other proteins
Inventor: WALLACH DAVID
IPC: A61K31/711
Filed: 1999-03-18
Abstract: 1. A DNA sequence encoding a RIP-associated protein (RAP-2), isoforms, fragments or analogs thereof including a sequence on Fig. 1, said RAP-2, isoforms, fragments or analogs thereof capable of binding to RIP or capable of modulating or mediating the intracellular activity of RIP. 2. A DNA sequence according to claim 1 comprising a sequence depicted on Fig. 2. 3. A DNA sequence according to claim 2 encoding RAP-2, isoforms, fragments or analogs thereof comprising at least a moiety of a sequence depicted in Fig.3. 4. A replicable expression vehicle comprising a DNA sequence according to any one of claims 1-3, optionally operatively linked with control sequences, promoters or other DNA sequences allowing expression in the correct orientation. 5. A replicable expression vehicle comprising a DNA sequence according to claim 4 capable of being expressed in a eukaryotic host cell. 6. A replicable expression vehicle according to claim 4 capable of being expressed in a prokaryotic host cell. 7. Transformed eukaryotic or prokaryotic host cells containing a replicable expression vehicle according to any one of claims 4-6. 8. A RAP-2 protein, isoform, fragment, functional analogs or derivatives thereof encoded by a DNA sequence according to any one of claims 1-3, said protein, isoform, fragment, analogs and derivatives thereof being capable of binding to RIP. 9. A RAP-2 protein according to claim 8 capable of modulating or mediating the intracellular activity of RIP in inflammation, cell survival or cell death pathways, in which RIP is involved directly, or indirectly by association with other intracellular modulators or mediators of these pathways. 10. A RAP-2 protein, isoform, fragment, analogs and derivatives thereof according to claim 8, wherein said protein, isoform, analogs, fragments and derivatives have at least part of the amino acid sequence depicted in Fig. 3. 11. A method for producing the RAP-2 protein, isoform, fragment, analogs or derivatives thereof according to any one of claims 8-10 comprising growing the transformed host cells according to claim 6 under conditions suitable for the expression of said protein, isoform, analog, fragment or derivative, effecting post-translational modifications as necessary for obtaining of said protein, fragments, analogs or derivatives and isolating said expressed protein, fragments, analogs or derivatives. 12. Antibodies or active fragments or derivatives thereof, specific for a RAP-2 protein, isoform, fragment, analog or derivative according to any one of claims 8-10. 13. A method for the modulation or mediation of the RIP modulated/mediated intracellular effects on the inflammation, cell death or cell survival pathways in which RIP is involved directly, or indirectly via other modulators/mediators of these pathways, comprising treating said cells with one or more RAP-2 proteins, isoforms, analogs, fragments or derivatives according to any one of claims 8-10 capable of binding to RIP and modulating or mediating said intracellular activity of RIP, wherein said treating of said cells comprises introducing into said cells said one or more proteins, isoforms, analogs, fragments or derivatives in a form suitable for intracellular introduction thereof, or introducing into said cells a DNA sequence encoding said one or more proteins, isoforms, analogs, fragments or derivatives in the form of a suitable vector carrying said sequence, said vector being capable of effecting the insertion of said sequence into said cells in a way that said sequence is expressed in said cells. 14. A method for the modulation of the RIP modulated/mediated effect on cells according to claim 13, wherein said treating of cells comprises introducing into said cells a DNA sequence encoding said RAP-2 protein, isoforms, analogs, fragments or derivatives in the form of a suitable vector carrying said sequence, said vector being capable of effecting the insertion of said sequence into said cells in a way that said sequence is expressed in said cells. 15. A method according to claim 13 or 14, wherein said treating of said cells is effected by transfection of said cells with a recombinant animal virus vector comprising the steps of: (a) constructing a recombinant animal virus vector carrying a sequence encoding a viral surface protein (ligand) that is capable of binding to a specific cell surface receptor on the surface of said cells to be treated and a second sequence encoding a protein selected from the RAP-2 protein, isoforms, analogs, fragments and derivatives according to any one of claims 11-13, that when expressed in said cells is capable of modulating/mediating the activity of RIP ; and (b) infecting said cells with said vector of (a). 16. A method for modulating RIP modulated/mediated effect on cells comprising treating said cells with antibodies or active fragments or derivatives thereof, according to claim 12, said treating being by application of a suitable composition containing said antibodies, active fragments or derivatives thereof to said cells, wherein when the RAP-2 protein or portions thereof of said cells are exposed on the extracellular surface, said composition is formulated for extracellular application, and when said RAP-2 proteins are intracellular said composition is formulated for intracellular application. 17. A method for modulating the RIP modulated/mediated effect on cells comprising treating said cells with an oligonucleotide sequence encoding an antisense sequence for at least part of the DNA sequence encoding a RAP-2 protein according to any one of claims 1-3, said oligonucleotide sequence being capable of blocking the expression of the RAP-2 protein. 18. A method according to claim 17, wherein said oligonucleotide sequence is introduced to said cells via a virus of claim 15. 19. A method for treating tumor cells or HIV-infected cells or other diseased cells comprising: (a) constructing a recombinant animal virus vector carrying a sequence encoding a viral surface protein capable of binding to a specific tumor cell surface receptor or HIV-infected cell surface receptor or receptor carried by other diseased cells and a sequence encoding a protein selected from the RAP-2 protein, isoform, analogs, fragments and derivatives according to any one of claims 8-10, that when expressed in said tumor, HIV-infected or other diseased cell is capable of enhancing the RIP modulated/mediated direct or indirect killing of said cell; and (b) infecting said tumor or HIV-infected cells or other diseased cells with said vector of (a). 20. A method for modulating the RIP effect on cells comprising applying the ribozyme procedure in which a vector encoding a ribozyme sequence capable of interacting with a cellular mRNA sequence encoding a RAP-2 protein according to any one of claims 8-10, is introduced into said cells in a form that permits expression of said ribozyme sequence in said cells, and wherein when said ribozyme sequence is expressed in said cells it interacts with said cellular mRNA sequence and cleaves said mRNA sequence resulting in the inhibition of expression of said RAP-2 protein in said cells. 21. A method according to any one of claims 13-20, wherein said protein is at least one of the RAP-2 isoforms, analogs, fragments and derivatives thereof. 22. A pharmaceutical composition for the modulation of the RIP effect on cells comprising as active ingredient at least one RAP-2 protein, according to any one of claims 8-10, its biologically active fragments, analogs, derivatives or mixtures thereof. 23. A pharmaceutical composition for modulating the RIP effect on cells comprising as active ingredient a recombinant animal virus vector encoding a protein capable of binding a cell surface receptor and encoding at least one RAP-2 protein, isoform, active fragments or analogs, according to any one of claims 8-10. 24. A pharmaceutical composition for modulating the RIP effect on cells comprising as active ingredient an oligonucleotide sequence encoding an anti-sense sequence of the RAP-2 protein DNA sequence according to any one of claims 1-3. 25. A method of modulating processes modulated/mediated by RIP directly or indirectly comprising treating cells with one or more RAP-2 proteins, isoforms, analogs, fragments or derivatives according to any one of claims 8-10 capable of binding to RIP, wherein said treating of said cells comprises introducing into said cells said one or more proteins, isoforms, analogs, fragments or derivatives in a form suitable for intracellular introduction thereof, or introducing into said cells a DNA sequence encoding said one or more proteins, isoforms, analogs, fragments or derivatives in the form of a suitable vector carrying said sequence, said vector being capable of effecting the insertion of said sequence into said cells in a way that said sequence is expressed in said cells. 26. A method of modulating processes that are mediated/modulated by RIP directly or indirectly and which include the inhibition of NF-KB and activation of JNK and p38 kinase comprising treating cells with one or more RAP-2 proteins, isoforms, analogs, fragments or derivatives according to any one of claims 8-10, wherein said treating of cells comprises introducing into said cells said one or more proteins, isoforms, analogs, fragments or derivatives in a form suitable for intracellular introduction thereof, or introducing into said cells a DNA sequence encoding said one or more proteins, isoforms, analogs, fragments or derivatives in the form of a suitable vector carrying said sequence, said vector being capable of effecting the insertion of said sequence into said cells in a way that said sequence is expressed in said cells. 27. A fragment according to any one of claims 8-10 being a peptide.