Chemistry: natural resins or derivatives; peptides or proteins; – Peptides of 3 to 100 amino acid residues – 25 or more amino acid residues in defined sequence
Reexamination Certificate
1999-02-25
2001-07-24
Eyler, Yvonne (Department: 1646)
Chemistry: natural resins or derivatives; peptides or proteins;
Peptides of 3 to 100 amino acid residues
25 or more amino acid residues in defined sequence
C530S350000, C435S007100
Reexamination Certificate
active
06265538
ABSTRACT:
Throughout this application various publications are referenced. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.
FIELD OF INVENTION
The present invention relates generally to the field of NF-&kgr;B-dependent activation to regulate immune, inflammatory, and anti-apoptotic responses. In particular, the present invention describes an inhibitor of serine/threonine kinase signaling that regulates NF-&kgr;B-dependent activation, thereby regulating immune, inflammatory, and anti-apoptotic responses. The present invention describes several mutant proteins of the NF-&kgr;B-inducing kinase (NIK) that prevent activation of a downstream kinase, I&kgr;B-specific kinase (IKK). One embodiment of the present invention describes kinase-deficient mutant NIK proteins that inhibits phosphorylation and activation of IKK. Another embodiment of the present invention describes an C-terminal NIK peptide protein that effectively interacts with IKK, and blocks IKK activation by inhibiting NIK/IKK association. Yet another embodiment of the present invention describes a method for inhibiting NF-&kgr;B-dependent immune and inflammatory responses using the NIK mutant proteins.
BACKGROUND OF THE INVENTION
TNF&agr; (tumor necrosis factor alpha) binding at the plasma membrane induces intracellular signaling events that translate into the induction of NF-&kgr;B in the nucleus. The eukaryotic NF-&kgr;B/Rel (nuclear factor-&kgr;B/Rel) family of transcription factors plays an essential role in the regulation of inflammatory, immune, and apoptotic responses (Bacuerle and Baltimore 1996; Baldwin 1996; Verma et al 1995). One of the distinguishing characteristics of the NF-&kgr;B/Rel transcription factor is its post-translational regulation through interactions with cytoplasmic inhibitory proteins termed I&kgr;B (inhibitor-&kgr;B). NF-&kgr;B corresponds to an inducible eukaryotic transcription factor complex that is negatively regulated in resting cells, by its physical assembly with a family of cytoplasmic ankyrin-rich I&kgr;B inhibitors (Bacuerle and Baltimore 1996; Baldwin 1996; Verma et al 1995). Stimulation of cells with various pro-inflammatory cytokines, including TNF&agr;, induces nuclear NF-&kgr;B expression. The TNF&agr;-signaling pathway is complex and involves recruitment of at least three adapter proteins, TRADD (TNF-R1 associated death domain protein) and TRAF-2 (TNF-receptor-associated factor)-2, and the serine/threonine kinase RIP (receptor interacting protein) to the cytoplasmic tail of the type 1 TNF receptor (Hsu et al Immunity 1996; Hsu et al Cell 1996) (FIG. 1). In turn, the recruitment of these factors promotes activation of the downstream NIK (Malinin et al 1997) and IKK&agr; and IKK&bgr; (I&kgr;B-specific kinases) (DiDonato et al 1997; Mercurio et al 1997; Regnier et al 1997; Woronicz et al 1997; Zandi et al 1997). The activated IKK&agr; and IKK&bgr; directly phosphorylates the two N-terminal regulatory serines within I&kgr;B&agr;, triggering ubiquitination and rapid degradation of this inhibitor in the 26S proteasome (Bacuerle and Baltimore 1996; Baldwin 1996; Verma et al 1995). Degradation of I&kgr;B&agr; unmasks the nuclear localization signal on NF-&kgr;B, allowing the NF-&kgr;B to translocate to the nucleus where it engages cognate &kgr;B enhancer elements and activates the transcription of various &kgr;B-dependent genes involved in inflammatory, immune, and anti-apoptotic responses.
IL-1, a second proinflammatory cytokine, acts in a manner similar to TNF&agr;. IL-1 binding to its receptor recruits the MyD88 and TRAF-6 adapter proteins and (IRAK), a serine-threonine kinase (Cao et al 1996; Muzio et al 1997). Like TRAF-2, TRAF-6 interacts with NIK. Thus, the TNF&agr; and IL-1 signaling pathways converge at the level of NIK. The present invention provides new insights into the molecular basis for NIK regulation of NF-&kgr;B-dependent gene expression, and provides novel methods for modulating NF-&kgr;B-dependent immune, inflammatory, and anti-apoptotic responses.
SUMMARY OF THE INVENTION
The present invention provides a molecular mechanism to interfere with cytokine induction of NF-&kgr;B-dependent immune, inflammatory, and anti apoptotic responses, through interference with the assembly of NIK and the IKK proteins. A relatively small region of NIK selectively impairs the NIK-IKK interaction. The present invention provides a novel and highly specific method for modulating NF-&kgr;B-dependent immune and inflammatory responses, based on interruption of the critical protein-protein interaction of NIK and IKK. The present invention provides methods for inhibiting NF-&kgr;B-dependent gene expression, using mutant NIK proteins. One embodiment of the present invention provides kinase-deficient NIK mutant proteins that inhibit activation of IKK. Another embodiment of the invention provides N-terminus NIK mutant proteins that bind IKK, thus inhibiting NIK/IKK interaction.
REFERENCES:
patent: WO 9737016 (1997-10-01), None
Zandi, Ebrahim et al., “T&kgr;B Kinase Complex (IKK) Contains Two Kinase Subunits, IKK&agr; and IKK&bgr;, Necessary for I&kgr;B Phosphorylation and NF-&kgr;B Activation,”Cell, Oct. 17, 1997, 91:243-52. (Exhibit 30).
Zheng, Chao-Feng and Kun-Liang Guan, “Activation of MEK Family Kinases Requires Phosphorylation of Two Conserved Ser/Thr Residues,”The EMBO Journal, 1994, 13:1123-31. (Exhibit 31).
Lee, Frank S. et al., “Activationof the I&kgr;B&agr; Kinase Complex by MEKK1, a Kinase of the JNK Pathway,”Cell, Jan. 24, 1997, 88:213-22. (Exhibit 14).
Liu, Zheng-gang et al., “Dissection of TNF Receptor 1 Effector Functions: JNK Activation is not Linked to Apoptosis While NF-&kgr;B Activation Prevents Cell Death,”Cell, Nov. 1, 1996, 87:565-76. (Exhibit 15).
Malinin, Nikolai L. et al., “MAP3K-Related Kinase Involved in NF-&kgr;B Induction by TNF, CD95 and IL-1,”Nature, Feb. 6, 1997, 385:540-4. (Exhibit 16).
Mercurio, Frank et al, “IKK-1 and IKK-2: Cytokine-Activated I&kgr;B Kinases Essential for NF-&kgr;B Activation,”Science, Oct. 31, 1997, 278:860-6. (Exhibit 17).
Muzio, Marta et al., “IRAK (Pelle) Family Member IRAK-2 and MyD88 as Proximal Mediators of IL-1 Signaling,”Science, Nov. 28, 1997, 278:1612-5. (Exhibit 18).
Perona, Rosario et al., “Activation of the Nuclear Factor-&kgr;B by Rho, CDC42 and Rac-1 Proteins,”Genes and Development, Feb. 15, 1997, 11:463-75. (Exhibit 19).
Regnier, Catherine H. et al., “Identification and Characterization of an I&kgr;B Kinase,”Cell, Jul. 25, 1997, 90:373-83. (Exhibit 20).
Siow, Yaw L. et al., “Identification of Two Essential Phosphorylated Threonine Residues in the Catalytic Domain of Mekk1 Indirect Activation by Pak3 and Protein Kinase C,”The Journal of Biological Chemistry, Mar. 21, 1997, 272:7586-94. (Exhibit 21).
Smith, Matthew R. and Warner C. Greene, “Molecular Biology of the Type Human T-Cell Leukemia Virus (HTLV-I) and Adult T-Cell Leukemia,”The Journal of Clinical Investigation, Mar. 1991, 87:761-6. (Exhibit 22).
Song, Ho Yeong et al., “Tumor Necrosis Factor (TNF)-Mediated Kinase Cascades: Bifurcation of Nuclear Factor-&kgr;B and c-jun N-Terminal Kinase (JNK/SAPK) Pathways at TNF Receptor-Associated Factor 2,”Proc. Natl. Acad. Sci. USA, Sep. 1997, 94:9792-6. (Exhibit 23).
Sulciner, David J. et al, “rac1 Regulates a Cytokine-Stimulated, Redox-Dependent Pathway Necessary of NF-&kgr;B Activation,”Molecular and Cellular Biology, Dec. 1996, 16:7115-7121. (Exhibit 24).
Sun, Shao-Cong et al., “Both Amino- and Carboxyl- Terminal Sequences Within I&kgr;B&agr; Regulate Its Inducible Degradation,”Molecular and Cellular Biology, Mar. 1996, 16:1058-65. (Exhibit 25).
Sun, Shao-Cong et al., “Human T-Cell Leukemia Virus Type I Tax Activation of NF-&kgr;B/Rel Involves Phosphorylation and Degradation of I&kgr;B&agr; and RelA (p65)-Mediated Induction of the c-rel Gene,”Molecular and Cellular Biology, Nov. 1994, 14:7377-84. (Exhibit 26).
Verma, Inder M. et al., “Rel/NF-&kgr;B Family: Intimate Tales of Associati
Gelezuinas Romas
Greene Warner C.
Lin Xin
Andres Janet L
Eyler Yvonne
Mandel & Adriano
The Regents of the University of California
LandOfFree
Inhibitor of the inflammatory response induced by the TNFA... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Inhibitor of the inflammatory response induced by the TNFA..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Inhibitor of the inflammatory response induced by the TNFA... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2540702