Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
1999-07-30
2001-11-06
Wang, Andrew (Department: 1635)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S091100, C435S325000, C435S375000, C514S04400A, C536S023100, C536S024500
Reexamination Certificate
active
06312900
ABSTRACT:
FIELD OF THE INVENTION
The present invention provides compositions and methods for modulating levels of the c-fos and c-jun genes, which encode the c-Fos and c-Jun subunits of AP-1, respectively. In vivo, AP-1, or transcription factor activating protein 1, is a heterogenous mixture of heterodimers of several related protein subunits including, in addition to c-Fos and c-Jun, FosB, Fra-1, Fra-2, c-Jun, JunB, JunD, etc. (The FOS and JUN Families of Proteins, Angel and Herrlich, eds., CRC Press, Boca Raton, Fla., 1994). AP-1 has been implicated in abnormal cell proliferation and tumor formation, events that thus might be controlled by modulating the expression of c-fos and/or c-jun. The invention is further directed to therapeutic, diagnostic, and research based reagents and methods for evaluating and treating disease states or disorders which result from and/or respond positively to modulation of one or more AP-1 subunits. Such disease states and disorders include those involving the hyperproliferation of cells such as, e.g., a tumor (neoplasm) or malignant cancer. Inhibition of AP-1-mediated hyperproliferation of cells, and corresponding prophylactic, palliative and therapeutic effects result from treatment with the oligonucleotides of the invention.
BACKGROUND OF THE INVENTION
Transcription factors play a central role in the expression of specific genes upon stimulation by extracellular signals, thereby regulating a complex array of biological processes. Members of the family of transcription factors termed AP-1 (activating protein-1) alter gene expression in response to growth factors, cytokines, tumor promoters, carcinogens and increased expression of certain oncogenes. Growth factors and cytokines exert their function by binding to specific cell surface receptors. Receptor occupancy triggers a signal transduction cascade to the nucleus. In this cascade, transcription factors such as AP-1 execute long term responses to the extracellular factors by modulating gene expression. Such changes in cellular gene expression lead to DNA synthesis, and eventually the formation of differentiated derivatives (Angel and Karin,
Biochim. Biophys. Acta
, 1991, 1072, 129).
In general terms, AP-1 denotes one member of a family of related heterodimeric transcription factor complexes found in eukaryotic cells or viruses. However, as used herein, “AP-1” specifically refers to the heterodimer formed of c-Fos and c-Jun (Angel and Herrlich, Chapter 1, and Schuermann, Chapter 2 in:
The FOS and JUN Families of Proteins
, Angel and Herrlich, eds., pp. 3-35, CRC Press, Boca Raton, Fla., 1994; Bohmann et al.,
Science
, 1987, 238, 1386; Angel et al.,
Nature
, 1988, 332, 166). These two proteins are products of the c-fos and c-jun proto-oncogenes, respectively. Repression of expression of either c-fos or c-jun, or of both proto-oncogenes, and the resultant inhibition of the formation of c-Fos and c-Jun proteins, is desirable for the inhibition of cell proliferation, tumor formation and tumor growth.
The phosphorylation of proteins plays a key role in the transduction of extracellular signals into the cell. Mitogen-activated protein (MAP) kinases, enzymes which effect such phosphorylations are targets for the action of growth factors, hormones, and other agents involved in cellular metabolism, proliferation and differentiation (Cobb et al.,
J. Biol. Chem
., 1995, 270, 14843). MAP kinases are themselves activated by phosphorylation catalyzed by, e.g., receptor tyrosine kinases, G protein-coupled receptors, protein kinase C (PKC), and the apparently MAP kinase dedicated kinases MEK1 and MEK2. MAP kinases include, but are not limited to, ERK1, ERK2, two isoforms of ERK3, ERK4 (ERK stands for “extracellular signal-regulated protein kinase), Jun N-terminal kinases/stress-activated protein kinases (JNKs/SAPKs), p38/HOG1, p57 MAP kinases, MKK3 (MAP kinase kinase 3) and MKK4 (MAP kinase kinase 4, also known as SAPK/ERK kinase (SEK) or JNK kinase (JNKK)) (Cobb et al.,
J. Biol. Chem
., 1995, 270, 14843, and references cited therein). In general, MAP kinases are involved in a variety of signal transduction pathways (sometimes overlapping and sometimes parallel) that function to convey extracellular stimuli to protooncogene products to modulate cellular proliferation and/or differentiation.
One of the signal transduction pathways involves the MAP kinases Jun kinase 1 and Jun kinase 2 which are responsible for the phosphorylation of specific sites (Serine 63 and Serine 73) on c-Jun. Phosphorylation of these sites potentiates the ability of AP-1 to activate transcription (Binetruy et al.,
Nature
, 1991, 351, 122; Smeal et al.,
Nature
, 1991, 354, 494). At least one human leukemia oncogene has been shown to enhance Jun N-terminal Kinase (JNK) function (Raitano et al.,
Proc. Natl. Acad. Sci
. (
USA
), 1995, 92, 11746), thus indirectly demonstrating a role for AP-1 in cellular hyperproliferation and tumorigenesis. Cellular hyperproliferation in an animal can have several outcomes. Hyperproliferating cells might be attacked and killed by the animal's immune system before a tumor can form., Tumors are abnormal growths resulting from the hyperproliferation of cells. Cells that proliferate to excess but stay put form benign tumors, which can typically be removed by local surgery. In contrast, malignant tumors or cancers comprise cells that are capable of undergoing metastasis, i.e., a process by which hyperproliferative cells spread to, and secure themselves within, other parts of the body via the circulatory or lymphatic system (see, generally, Chapter 16 In:
Molecular Biology of the Cell
, Alberts et al., eds., pp. 891-950, Garland Publishing, Inc., New York, 1983). Using the oligonucleotides of the invention, it has surprisingly been discovered that several genes encoding enzymes required for metastasis are positively regulated by AP-1. Accordingly, inhibition of expression of c-fos and/or c-jun serves as a means to modulate the metastasis of malignant tumors. A method of modulating one or more metastatic events using the oligonucleotides of the invention is thus herein provided.
RELEVANT ART
Soprano et al. (
Ann. N. Y. Acad. Sci
., 1992, 660, 231) have used antisense oligodeoxynucleotides targeted to c-jun mRNA to study their ability to inhibit DNA synthesis and cell division.
Liu et al. (
Ann. Neurol
., 1994, 36, 566) describe the suppression of c-fos by intraventricular infusion of an antisense oligodeoxynucleotide targeted to c-fos mRNA.
Chen et al. (
Cancer Lett
., 1994, 85, 119) describe repression of c-jun expression by antisense oligodeoxynucleotides resulting in the inhibition of cell proliferation in E5a transformed cells.
Gillardon et al. describe the topical application of c-fos antisense oligodeoxynucleotides to the rat spinal cord (
Eur. J. Neurosci
., 1994, 6, 880) ultraviolet (UV)-exposed rat eyes (
British J. Ophthal
., 1995, 79, 277) and UV-irradiated rat skin (
Carcinogenesis
, 1995, 16, 1853).
U.S. Pat. No. 5,602,156, which issued Feb. 11, 1997, to Kohn et al., discloses non-oligonucleotide compositions and methods for inhibiting the expression of two metalloproteinases, MMP-1 and MMP-2.
International Publication Number WO 95/02051, published Jan. 19, 1995, discloses antisense oligonucleotides targeted to the mRNA of c-fos and c-jun.
International Publication Number WO 95/03323, published Feb. 2, 1995, discloses antisense nucleic acids which are complementary to the polynucleotide encoding a polypeptide which is capable of phosphorylating the c-jun N-terminal activation domain. Also provided are methods for treating a cell proliferative disorder associated with said polypeptide.
International Publication Number WO 95/03324, published Feb. 2, 1995, describes a polypeptide which phosphorylates the c-jun N-terminal activation domain. This publication also discloses a polynucleotide sequence encoding the polypeptide.
To date, there are no known therapeutic agents which effectively inhibit gene expression of c-fos and/or c-jun. Furthermore, there are to date no known therapeutic agents that mod
Baker Brenda
Dean Nicholas M.
McKay Robert
Miraglia Loren
ISIS Pharmaceuticals Inc.
Licata & Tyrrell P.C.
Wang Andrew
LandOfFree
Antisense oligonucleotide compositions and methods for the... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Antisense oligonucleotide compositions and methods for the..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Antisense oligonucleotide compositions and methods for the... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2600849