Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Antisense oligonucleotide compositions and methods for the modulation of activating protein 1

an activating protein and anti-oligonucleotide technology, applied in the direction of biocide, peptide/protein ingredients, genetic material ingredients, etc., can solve the problems of no known therapeutic agents that effectively inhibit the gene expression of c-fos, no known therapeutic agents that modulate the metastasis of malignant cells, and no known therapeutic agents for modulating the function of cells

Inactive Publication Date: 2002-04-04
DEAN NICHOLAS M +3
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0017] In accordance with the present invention, oligonucleotides are provided which specifically hybridize with nucleic acids encoding c-Fos or c-Jun. Certain oligonucleotides of the invention are designed to bind either directly to mRNA transcribed from, or to a selected DNA portion of, the respective gene, thereby modulating the amount of protein translated from a c-fos or c-jun mRNA and / or the amount of mRNA transcribed from a c-fos or c-jun gene, respectively. Such modulation can, in turn, effect the modulation of enzymes and cellular processes involved in the metastasis of malignant cells.

Problems solved by technology

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 modulate the metastasis of malignant cells.
Such hybridization with mRNA interferes with the normal role of mRNA and causes a modulation of its function in cells.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

example 2

[0085] Screening for Oligonucleotides that Modulate mRNA Expression of the AP-1 Subunits c-fos and c-jun

[0086] In order to evaluate the activity of potential c-fos and c-jun modulating oligonucleotides, A549 cells were grown in T-75 flasks until 80-90% confluent. (Cell line A549 is available from, inter alia, the American Type Culture Collection, Rockville, Md., as ATCC No. CCL-185.) At this time, the cells were washed twice with 10 mL of media (DMEM), followed by the addition of 5 mL of DMEM containing 20 .mu.g / mL of LIPOFECTIN.TM. (i.e., DOTMA / DOPE (N-[1-(2,3-dioleyloxy) propyl]-N,N,N-triethylammonium chloride / dioleoylphosphatidyl ethanolamine)). The oligonucleotides were added from a 10 .mu.M stock solution to a final concentration of 400 nM, and the two solutions were mixed by swirling the flasks. After 4 hours at 37.degree. C., the medium was replaced with DMEM containing 10% serum. At this point, 1 .mu.M 12-O-tetradecanoylphorbol 13-acetate (TPA) was added to induce expression...

example 3

[0089] Dose Response and Specificity of Oligonucleotides a Targeted to AP-1 Subunits

[0090] Dose-response experiments were performed at different oligonucleotide concentrations to determine the potency (i.e., ability to decrease expression of the appropriate mRNA target) of the most active compounds identified in the initial screen (Tables 3 and 4). The decreases in target mRNA expression effected by ISIS 10582 ( c-jun) and ISIS 10639 (c-fos) are dose-dependent, as shown in Tables 3 and 4, respectively.

3TABLE 3 5 / 28 Dose-Response to Oligonucleotides Targeted to c-jun OLIGONUCLEOTIDE c-jun mRNA LEVELS TREATMENT CONCENTRATION (% CONTROL) None (basal level) --31 Control* -- 100 TPA + ISIS 10582 50 nM 72 TPA + ISIS 10582 100 nM 45.5 TPA + ISIS 10582 200 nM 29.5 TPA + ISIS 10582 400 nM 16 *Control is TPA induction, at 1 hour, in A549 cells.

[0091]

4TABLE 4 Dose-Response to Oligonucleotides Targeted to c-fos OLIGONUCLEOTIDE c-fos mRNA LEVELS TREATMENT CONCENTRATION (% CONTROL) None (basal le...

example 4

[0095] Effect of Oligonucleotides Targeted to an AP-1 Subunit on Human Tumor Growth in Nude Mice

[0096] In order to evaluate the in vivo activity of c-fos oligonucleotides, 25 mg of tumor fragments of A549 tumors were implanted subcutaneously in nude mice (n=6). ISIS 10639 was administered daily, i.v., for three weeks. The oligonucleotide dosage was 25 mg / kg. Tumor size was recorded weekly, and the results are shown in Table 6. A substantial reduction in tumor growth rate was obtained upon treatment with ISIS 10639. By day 34, saline-treated tumors were 0.56.+-.0.12 g by weight, while tumors treated with ISIS 10639 were 0.31.+-.0.1 g by weight.

6TABLE 6 Response of Transplanted Tumors in Mice to Oligonucleotides Targeted to c-jun Mean Tumor Treatment / Time Weight (g) Std. Dev. Std. Error Saline: Day 17 0.113 0.033 0.014 Day 20 0.177 0.045 0.019 Day 27 0.272 0.086 0.035 Day 34 0.560 0.293 0.120 ISIS 10639: Day 17 0.105 0.035 0.014 Day 20 0.138 0.074 0.030 Day 27 0.225 0.070 0.028 Day 34...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
pHaaaaaaaaaa
covalent linkagesaaaaaaaaaa
covalent linkageaaaaaaaaaa
Login to View More

Abstract

Compositions and methods for the treatment and diagnosis of diseases or disorders amenable to treatment through modulation of Activating Protein 1 (AP-1) expression are provided. In accordance with various embodiments of the present invention, oligonucleotides are provided which are specifically hybridizable with c-fos or c-jun, the genes encoding c-Fos or c-Jun, respectively. In a preferred embodiment, a method of modulating the metastasis of malignant tumors via modulation of one or more of the AP-1 subunits is provided; this method can be effected using the oligonucleotides of the invention or any other agent which modulates AP-1 or AP-1 mediated transcription.

Description

[0001] This application is a divisional of U.S. patent application Ser. No. 09 / 364,416 filed Jul. 30, 1999, which is a continuation of U.S. patent application Ser. No. 08 / 837,201 filed Apr. 14, 1997, now issued as U.S. Pat. No. 5,985,558.[0002] 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, 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 ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/00C12N15/113
CPCA61K38/00C12N15/1135C12N2310/335C12N2310/334C12N2310/33C12N2310/322C12N2310/321C12N2310/315C12N2310/3181C12N2310/3525C12N2310/3521C12N2310/3527
Inventor DEAN, NICHOLAS M.MCKAY, ROBERTMIRAGLIA, LORENBAKER, BRENDA
Owner DEAN NICHOLAS M
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com