Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
1994-03-18
2004-02-10
Priebe, Scott D. (Department: 1632)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S004000, C435S029000, C435S375000, C435S455000, C536S024500
Reexamination Certificate
active
06689560
ABSTRACT:
BACKGROUND OF THE INVENTION
Field of the Invention
The present invention relates, in general, to methods of inducing a therapeutic effect. In particular, the present invention relates to therapeutic uses of Raf protein kinases.
Background Information
Raf serine- and threonine-specific protein kinases are cytosolic enzymes that stimulate cell growth in a variety of cell systems (Rapp, U. R., et al. (1988) In The oncogene handbook; T. Curran, E. P. Reddy, and A. Skalka (ed.) Elsevier Science Publishers; The Netherlands, pp.213-253; Rapp, U. R., et al. (1988) Cold Spring Harbor Sym. Quant. Biol. 53:173-184; Rapp, U.R., et al. (1990) In: Curr. Top. Microbiol. Immunol. Potter and Melchers (eds), Berlin, Springer-Verlag 166:129-139). Three isozymes have been characterized: c-Raf (Raf-1) (Bonner, T. I., et al. (1986)
Nucleic Acids Res.
14:1009-1015 see FIG.
9
). A-Raf (Beck, T. W., et al. (1987)
Nucleic Acids Res.
15:595-609 see FIG.
10
), and B-Raf (Ikawa, S., et al. (1988) Mol. Cell. Biol. 8:2651-2654; Sithanandam, G. et al. (1990) Oncogene 5:1775 see FIG.
11
). These enzymes differ in their expression in various tissues. Raf-1 is expressed in all organs and in all cell lines that have been examined, and A- and B-Raf are expressed in urogenital and brain tissues, respectively (Storm, S. M. (1990) Oncogene 5:345-351).
Raf genes are proto-oncogenes: they can initiate malignant transformation of cells when expressed in specifically altered forms. Genetic changes that lead to oncogenic activation generate a constitutively active protein kinase by removal or interference with an N-terminal negative regulatory domain of the protein (Heidecker, G., et al. (1990) Mol. Cell. Biol. 10:2503-2512; Rapp, U. R., et al. (1987) In Oncogenes and cancer S. A. Aaronson, J. Bishop, T. Sugimura, M. Terada, K. Toyoshima, and P. K. Vogt (ed.) Japan Scientific Press, Tokyo). Microinjection into NIH 3T3 cells of oncogenically activated but not wild-type versions of the Raf-protein prepared with
Escherichia coli
expression vectors results in morphological transformation and stimulates DNA synthesis (Rapp, U. R., et al. (1987) In Oncogenes and cancer; S. A. Aaronson, J. Bishop, T. Sugimura, M. Terada, K. Toyoshima, and P. K. Vogt (ed.). Japan Scientific Press, Tokyo; Smith, M. R., et al. (1990) Mol. Cell. Biol. 10:3828-3833). Thus, activated Raf-1 is an intracellular activator of cell growth. Raf-1 protein serine kinase is a candidate downstream effector of mitogen signal transduction, since Raf oncogenes overcome growth arrest resulting from a block of cellular ras activity due either to a cellular mutation (ras revertant cells) or microinjection of anti-ras antibodies (Rapp, U. R., et al. (1988) In The Oncogene Handbook, T. Curran, E. P. Reddy, and A. Skalka (ed.), Elsevier Science Publishers; The Netherlands, pp.213-253; Smith, M. R., et al. (1986) Nature (London) 320:540-543).
c-Ras function is required for transformation by a variety of membrane-bound oncogenes and for growth stimulation by mitogens contained in serum (Smith, M. R., et al. (1986) Nature (London) 320:540-543). Raf-1 protein merine kinase activity is regulated by mitogens via phosphorylation (Morrison, D. K., et al. (1989) Cell 58:648-657), which also effects subcellular distribution (Olah, Z., et al. (1991) Exp. Brain Res.84:403; Rapp, U. R., et al. (1988) Cold Spring Harbor Sym. Quant. Biol. 53:173-184).
Raf-1 activating growth factors include platelet-derived growth factor (PDGF) (Morrison, D. K., et al. (1988) Proc. Natl. Acad. Sci. USA 85:8855-8859) colony-stimulating factor 1 (Baccarini, M., et al. (1990) EMBO J. 9:3649-3657), insulin (Blackshear, P. J., et al. (1990) J. Biol. Chem. 265:12131-12134; Kovacina, K. S., et al. (1990) J. Biol. Chem. 265:12115-12118), epidermal growth factor (EGF) (Morrison, D. K., et al. (1988) Proc. Natl. Acad. Sci. USA 85:8855-8859), interleukin 2 (Turner, B. C. et al. (1991) Proc. Natl. Acad. Sci. USA 88:1227), and interleukin 3 and granulocyte-macrophage colony-stimulating factor (Carroll, M. P., et al (1990) J. Biol. Chem. 265:19812-19817). Upon mitogen treatment of cells, the transiently activated Raf-1 protein serine kinase translocates to the perinuclear area and the nucleus (Olah, Z., et al. (1991) Exp. Brain Res. 84:403; Rapp, U. R., et al. (1988) Cold Spring Harbor Sym. Quant. Biol. 53:173-184). Cells containing activated Raf are altered in their pattern of gene expression (Heidecker, G., et al. (1989) In Genes and signal transduction in multistage carcinogenesis, N. Colburn (ed.), Marcel Dekker, Inc., New York. pp. 339-374), and Raf oncogenes activate transcription from Ap-1/PEA3-dependent promoters in transient transfection assays (Jamal, S., et al. (1990) Science 344:463-466; Kaibuchi, K., et al. (1989) J. Biol. Chem. 264:20855-20858; Wasylyk, C., at al. (1989) Mol. Cell. Biol. 9:2247-2250).
There are at least two independent pathways for Raf-1 activation by extracellular mitogens: one involving protein kinase C (PKC) and a second initiated by protein tyrosine kinases (Blackshear, P. J., et al. (1990) J. Biol. Chem. 265:12131-12134; Kovacina, K. S., et al. (1990) J. Biol. Chem. 265:12115-12118; Morrison, D. K., et al. (1988) Proc. Natl. Acad. Sci. USA 85:8855-8859; Siegel, J. N., et al.(1990) J. Biol. Chem. 265:18472-18480; Turner, B. C. et al. (1991) Proc. Natl. Acad. Sci. USA 88:1227). In either case, activation involves Raf-1 protein phosphorylation. Raf-1 phosphorylation may be a consequence of a kinase cascade amplified by autophosphorylation or may be caused entirely by autophosphorylation initiated by binding of a putative activating ligand to the Raf-1 regulatory domain, analogous to PKC activation by diacylglycerol (Nishizuka, Y. (1986) Science 233:305-312).
SUMMARY OF THE INVENTION
It is a general object of this invention to provide a construct comprising a DNA segment comprising a Raf gene in an antisense orientation downstream of a promoter.
It is a specific object of this invention to provide a method of inhibiting Raf expression comprising expressing an antisense Raf gene in a cell such that said Raf expression is inhibited.
It is a further object of the invention to provide a method of inhibiting Raf kinase activity comprising replacing a serine or threonine amino acid within the Raf gene for an amino acid not susceptible to phosphorylation.
Further objects and advantages of the present invention will be clear from the description that follows.
REFERENCES:
patent: 4740463 (1988-04-01), Weinberg et al.
Orkin et al. , “Report and Recommendations of the panel to assess the NIH invesment in research on gene therapy”, issued by the National Institutes of Health, Dec. 1995.*
Ellis et al. (1990) “Phosphorylation of GAP and GAP-Associated Proteins by Transforming and Nitrogenic Tyrosine Kinases” Nature 343: 377-381.*
Cantley et al. (1991) “Oncogenes and Signal Transduction.” Cell 64: 281-302.*
Hunter et al. (1991) “Cooperation Between Oncogenes” Cell 64: 249-270.*
Ishikawa et al (1988) Identification of a Transforming Activity Suppressing Sequence in the C-Raf Cocogene 3 (6): 653-658.*
Soderling (1990) Protein Kinases: Regulation by Antoinhibitory Domains. J. Biol. Chem. 265 (4): 1823-1826.*
Heidecker et al. (1990) Mutational activation of C-Raf-1 and Definition of the Minimal Transforming Sequence. Mol. Cell. Biol. 10(6): 2503-2512.*
Feig et al. (1988) Inhibition of NIH 3T3 Cell Proliferation by a Mutant Raf Protein . . . Mol. Cell. Biol. 8(8): 3235-3243.*
App et al. “Epidermal Growth Factor (EGF) Stimulates Association and Kinase Activity of Raf-1 with the EGF Receptor” Mol. and Cell. Biol. 11(2):913-919, Feb. 1991.
Kolch et al. “Raf-1 protein kinase is required for growth of induced NIH/3T3 cells”Nature349:426-428, Jan. 31, 1991.
Rapp et al. “Oncogenes: Clinical Relevance”Haematol. and Blood Transf. 31:450-459, 1987(a).
Rapp et al. “Transformation byrafandmycOncogenes” inOncogenes and Cancer, Ed. Aaronson et al., pp. 55-74, 1987(b).
Bonner et al. “The Complete Coding Sequence of the HumanrafOncogene and the Corresponding Structure of the c-raf-1 gene”Nucl. Acids Res. 1
App Harald
Rapp Ulf R.
Storm Stephen M.
Needle & Rosenberg P.C.
Priebe Scott D.
The United States of America as represented by the Department of
LandOfFree
Raf protein kinase therapeutics does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Raf protein kinase therapeutics, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Raf protein kinase therapeutics will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3326258