Claims
- 1. A method for selectively inhibiting degradation of IκBα within a targeted collection of viable cells in-situ, said method comprising the steps of:
identifying a collection of cells comprising viable cells in-situ as a target for inhibiting IκBα degradation; providing means for effecting an introduction of at least one member selected from the group consisting of the PR-39 oligopeptide collective to the cytoplasm of said targeted collection of cells; introducing at least one member of the PR-39 oligopeptide collective to the cytoplasm of said targeted collection of cells using said effecting means; allowing said introduced PR-39 oligopeptide collective member to interact with such IκBα and proteasomes as are present within the cytoplasm of said targeted collection of cells whereby
(a) at least some of the proteasomes interact with said PR-39 oligopeptide collective member, (b) at least a part of the proteolytic activity mediated by said proteasomes becomes selectively altered by said interaction, and (c) the selectively altered proteolytic activity of said proteasomes results in a marked inhibition of IκBα degradation in-situ within the cytoplasm of said targeted collection of viable cells.
- 2. A method for decreasing the activity of NFκB transcription factor within a targeted collection of viable cells in-situ, said method comprising the steps of:
identifying a collection of cells comprising viable cells in-situ as a target for decreased NFκB activity; providing means for effecting an introduction of at least one member selected from the group consisting of the PR-39 oligopeptide collective to the cytoplasm of said targeted collection of cells; introducing at least one member of the PR-39 oligopeptide collective to the cytoplasm of said targeted collection of cells using said effecting means; allowing said introduced PR-39 oligopeptide collective member to interact with such IκBα and proteasomes as are present within the cytoplasm of said targeted collection of cells whereby
(a) at least some of the proteasomes interact with said PR-39 oligopeptide collective member, (b) at least a part of the proteolytic activity mediated by said proteasomes becomes selectively altered by said interaction, (c) the selectively altered proteolytic activity of said proteasomes results in a marked inhibition of IκBα degradation in-situ within the cytoplasm of said targeted collection of viable cells, and (d) said reduction of IκBα degradation results in a decrease in activity for such NFκB transcription factor as is present intracellularly.
- 3. A method for selective control of NFκB-dependent gene expression in-situ within a collection of viable cells, said method comprising the steps of:
identifying a collection of cells comprising viable cells in-situ as a target for controlling NFκB-dependent gene expression; providing means for effecting an introduction of at least one member selected from the group consisting of the PR-39 oligopeptide collective to the cytoplasm of said targeted collection of cells; introducing at least one member of the PR-39 oligopeptide collective to the cytoplasm of said targeted collection of cells using said effecting means; allowing said introduced PR-39 oligopeptide collective member to interact with such IκBα and proteasomes as are present within the cytoplasm of said targeted collection of cells whereby
(a) at least some of the proteasomes interact with the PR-39 oligopeptide collective member, (b) at least a part of the proteolytic activity mediated by said proteasomes becomes selectively altered by said interaction, (c) the selectively altered proteolytic activity of said proteasomes results in a marked reduction of IκBα degradation in-situ within the cytoplasm of said targeted collection of cells, (d) said reduction of IκBα degradation results in a decrease in activity for such NFκB transcription factor as is present intracellularly, and (e) NFκB-dependent gene expression is selectively controlled in said targeted collection of viable cells.
- 4. The method as recited in claim 1, 2 or 3 wherein said collection of viable cells includes at least one type of cell selected from the group consisting of endothelial cells, myocytes and myoblasts, fibrocytes and fibroblasts, epithelial cells, osteocytes and osteoblasts, neuronal cells and glial cells, erythrocuctes, leukocytes, and progenitor cells of all types.
- 5. The method as recited in claim 1, 2 or 3 wherein said collection of cells comprises at least one tissue selected from the group consisting of myocardium, skeletal muscle, smooth muscle, an artery, a vein, lung, brain, kidney, spleen, liver, gastrointestinal tissue, nerve tissue, limbs, and extremities.
- 6. The method as recited in claim 1, 2 or 3 wherein the means for an introduction of a PR-39 oligopeptide collective member include one selected from the group consisting of catheter-based introduction means, injection-based introduction means, infusion-based introduction means, localized intravascular introduction means, liposome-based introduction means, receptor-specific peptide introduction means, slow releasing means for peptide secretion in living cells and sequested organisms.
- 7. The method as recited in claim 1, 2 or 3 wherein the means for an introduction of a PR-39 oligopeptide collective member includes the DNA sequences coding for PR-39 oligopeptides of different sizes inserted in a suitable vector for transfection and subsequent expression of peptides within said cells.
- 8. The method as recited in claim 1, 2 or 3 wherein said method is practiced under in-vivo conditions.
- 9. The method as recited in claim 1, 2 or 3 wherein said method is practiced under in-vitro conditions.
- 10. A family of PR-39 derived oligopeptides whose members individually cause a selective inhibition of proteasome-mediated IκBα degradation in-situ after introduction intracellularly to a viable cell, each member of said oligopeptide family being:
a peptide less than 39 amino acid residues in length; at least partially homologous with the N-terminal amino acid residue sequence of the native PR-39 peptide; able to interact in-situ with such IκBα as is present within the cytoplasm of the cell; and able to inhibit markedly the degradation activity of proteasomes intracellularly as a consequence of said interaction with IκBα.
- 11. The PR-39 derived oligopeptide family as recited in claim 10 whose membership includes a peptide comprised of 15 amino acid residues whose sequence is Arg-Arg-Arg-Pro-Arg-Pro-Pro-Tyr-Leu-Pro-Arg-Pro-Arg-Pro-Pro.
- 12. The PR-39 derived oligopeptide family as recited in claim 10 whose membership includes a peptide comprised of 11 amino acid residues whose sequence is Arg-Arg-Arg-Pro-Arg-Pro-Pro-Tyr-Leu-Pro-Arg.
- 13. The PR-39 derived oligopeptide family as recited in claim 10 whose membership includes a peptide comprised of 8 amino acid residues whose sequence is Arg-Arg-Arg-Pro-Arg-Pro-Pro-Tyr.
CROSS REFERENCE
[0001] The present application is a Continuation-In-Part of prior pending U.S. patent applications Ser. Nos. 09/276,868 filed Mar. 26, 1999 and 09/426,011 filed Oct. 25, 1999.
RESEARCH SUPPORT
[0002] The research effort for the invention was supported in part by the National Institutes of Health, grants HL 53793 and HL 56993 (MS), DK 31396 (MS), GM51923 and GM 46147 (ALG), F32HL 10013 (RV), and in part by a grant from Chiron Corporation. The government has certain rights in the invention.
Divisions (1)
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Number |
Date |
Country |
Parent |
09474967 |
Dec 1999 |
US |
Child |
10391155 |
Mar 2003 |
US |
Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
09276868 |
Mar 1999 |
US |
Child |
09474967 |
Dec 1999 |
US |
Parent |
09426011 |
Oct 1999 |
US |
Child |
09474967 |
Dec 1999 |
US |