Drug – bio-affecting and body treating compositions – Lymphokine
Patent
1996-03-05
2000-06-27
Fitzgerald, David L.
Drug, bio-affecting and body treating compositions
Lymphokine
530350, 530351, 536 235, 435 695, 435 691, 4353201, 435325, 4352523, 43525411, A61K 3819, C07K 1452, C12N 1519
Patent
active
060803983
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates generally to certain proteins and to methods of improving the biological activity of certain proteins, and more specifically, chemokines.
BACKGROUND OF THE INVENTION
All the members of the intercrine or chemokine family are basic heparin-binding polypeptides which have four cysteine residues which form two disulfide bridges. All these proteins which have been functionally characterized appear to be involved in proinflammatory and/or restorative functions. As such, these molecules are anticipated to have therapeutic potential in bone marrow transplantation and the treatment of infections, cancer, myelopoietic dysfunction, graft versus host disease, and autoimmune diseases.
The chemokine family can be divided into two subfamilies depending upon their amino acid sequence and chromosomal location. The members of the .alpha. subfamily are termed the "C-X-C" subfamily because the first two cysteines are separated by only one amino acid. The human genes in this subfamily include IL-8, GRO/MGSA, and IP-10; murine counterparts include KC and macrophage inflammatory protein 2 (MIP-2). In the chemokine .beta. subfamily, the first two cysteines are in an adjacent position (the "C-C" subfamily). This subfamily includes human MCAF, LD-78, ACT-2, and RANTES. The murine counterparts are JE, TCA-3, MIP-1.alpha., and MIP-1.beta. [J. J. Oppenheim et al, Annu. Rev. Immunol., 9:617-648 (1991)].
The murine KC gene product [Oquendo et al, J. Biol. Chem., 264:4233 (1989)] is induced by platelet-derived growth factor (PDGF) and this is thought to be the murine homolog of the human MGSA/gro.alpha. gene (63.0% amino acid identity to mMIP-2). KC has been expressed in COS-1 cells to show that it encodes a secreted protein [Oquendo, cited above].
Two forms of MIP have been found in cultures of macrophage tumor cells from the mouse: MIP-1 and MIP-2. Murine MIP-2 (mMIP-2) is an inducible protein whose cDNA also has been cloned and sequenced [International Patent Application, Publication No. WO 90/02762 (Mar. 22, 1990)]. Murine MIP-2 has been shown to have potent chemotactic activity for human polymorphonuclear leukocytes (PMN), and to induce PMN degranulation of lysozyme but not of .beta.-glucuronidase [Wolpe et al, J. Exp. Med., 167:570 (1987)]. Further, mMIP-2 has been shown to have myelopoietic enhancing activities for CFU-GM [Broxmeyer et al, J. Exp. Med., 170:1583 (1989)]. The human counterpart of this factor was found to consist of two species, MIP-2.alpha. and MIP-2.beta., also termed gro-.beta. and gro-.gamma., respectively.
The cDNA and amino acid sequences of human gro-.beta. are provided in International Patent Application, Publication No. WO 92/00327 (Jan. 9, 1992); the cDNA and amino acid sequences of human gro-.gamma. are provided in International Patent Application, Publication No. WO 92/00326 (Jan. 9, 1992). Each of these sequences were predicted to encode a 73 amino acid mature protein.
MGSA or gro-.alpha. [Richmond et al, EMBO J., 7:2025 (1988)] is an autocrine growth factor with potent mitogenic activity secreted by human melanoma cells and is the product of the human gro gene [Anisowicz et al, Proc. Natl. Acad. Sci., 84:7188 (1987)].
There remains a need in the art for methods of enhancing the bioactivity of these mature proteins to enable their efficient use as therapeutic or pharmaceutical products, and to minimize the amounts of the proteins necessary to produce a therapeutic effect, thereby lowering toxicity.
SUMMARY OF THE INVENTION
In one aspect, the present invention provides a modified chemokine, which includes KC protein, human gro .alpha., gro-.beta., and gro-.gamma., which modified protein is characterized by truncation of between about 2 to about 8 amino acids at the amino terminus of the mature protein and by at least a log higher biological activity than the mature protein.
In another aspect, the present invention provides a modified chemokine which is characterized by truncation of between about 2 to about 10 amino acids at the carboxy terminus
REFERENCES:
patent: 5154921 (1992-10-01), Sager et al.
patent: 5459128 (1995-10-01), Rollins et al.
patent: 5703206 (1997-12-01), Wolpe
patent: 5739103 (1998-04-01), Rollins et al.
patent: 5854412 (1998-12-01), Rollins et al.
Proost, et al., "Identification of Novel Granulocyte Chemotactic Protein (GCP-2) from Human Tumor Cells", (1993), Journal of Immunology, vol. 150:3, pp. 1000-1010.
Cuenca, et al, "Characterization of GRO .alpha., .beta., and .gamma. expression in human colonic tumours: potential significance of cytokine involvement", (1992), Surgical Oncology, vol. 1, pp. 323-329.
Broxmeyer, et al., "Comparative Analysis of the Human Macrophage Inflammatory Protein Family of Cytokines (Chemokines) on Proliferation of Human Myeloid Progenitor Cells", (1993), Journal of Immunology, vol. 150:8, pp. 3448-3458.
Broxmeyer, et al., "Enhancing and Suppressing Effects of Recombinant Murine Macrophage Inflammatory Proteins on Colony Formation In Vitro by Bone Marrow Myeloid Progenitor Cells", (1990), Blood, vol. 76:6, pp. 1110-1116.
Bowie, et al., "Deciphering the Message in ProteinSequences: Tolerance to Amino Acid Substitutions", (1990), Science, vol. 247, pp. 1306-1310.
Watanabe, et al., "Rat Gro/Melanoma Growth-Stimulating Activity. Assessment of the Structure Responsible for Chemotactic Activity by Use of Its Fragments Prepared by Proteolysis and Chemical Synthesis", (1992), Cytokine, vol. 4:1, pp. 12-17.
Lord, et al., "Macrophage inflammatory protein: its characteristics, biological properties and role in the regulation of haemopoiesis", (1993), Intl. Journal of Hematology, vol. 57, pp. 197-206.
Sager, R., "Gro as a Cytokine", (1990), Prog. Leukocyte Biol., vol. 10 A, Oppenheim, J.J., et al., eds., Mol. Cell. Biol. of Cytokines (Proc. 2d Int'l Workshop on Cytokines, Dec. 10-14, 1989), pp. 327-332.
Wolpe, et al., "Macrophage inflammatory proteins 1 and 2: members of a novel superfamily of cytokines", (1989), The FASEB Journal, vol. 3, pp. 2565-2573.
Anisowicz, et al., "Constitutive overexpression of a growth-regulated gene in transformed Chinese hamster and human cells", (1987), Proc. Natl. Acad., vol. 84, pp. 7188-7192.
Richmond, et al., "Molecular characterization and chromosomal mapping of melanoma growth stimulatory activity, a growth factor structurally related to .beta.-thromboglobulin", (1988), The EMBO Journal, vol. 7:7, pp. 2025-2033.
M.Y. Stoeckle, "Post-transcriptional regulation of gro.alpha., .beta., .gamma., and IL-8 mRNAs by IL-1.beta.", (1991), Nucleic Acids Research, vol. 19, No. 4, pp. 917-920.
Sporn, et al., "Isolation of Adherence Specific cDNA Clones from a Monocyte cDNA Library", (1988), J. Leukocyte Biology, 44(4), p. 267.
Sporn, et al., "Monocyte Adherence Induces Novel Genes Sharing Homology With Mediators of Inflammation and Tissue Repair", (1989), J. Leukocyte Biology, 46(4), p. 328.
Sporn, et al., "Monocyte Adherence Results in Selective Induction of Novel Genes Sharing Homology With Mediators of Inflammation and Tissue Repair", (1990), J. Immunology, 144(11), pp. 4434-4441.
Haskill, et al., "Identification of three related human GRO genes encoding cytokine functions", (1990), Proc. Natl. Acad. Sci. USA, 87(19), pp. 7732-7736.
Anisowicz, et al, "Functional diversity of gro gene expression in human fibroblasts and mammary epithelial cells", (1988), Proc. Natl. Acad. Sci. USA, 87(19), pp. 9645-9649.
Trask, et al., "The Gro Gene as Growth Factor and Cytokine", (1990), J. Cell Biochem., Suppl. 0 (14 pt. B) p. 5.
Tekamp-Olson, et al., "Cloning and Characterization of cDNAs for Murine Macrophage Inflammatory Protein 2 and its Human Homologues", (1990), J. Exp. Med., 172, pp. 911-919.
Suggs, et al., "Use of synthetic oligonucleotides as hybridization probes: Isolation of cloned cDNA sequences for human .beta.2-microglobulin", (1981), Proc. Natl. Acad. Sci. USA, 78:11, pp. 6613-6617.
E. Brandt, et al., "A Novel Molecular variant of the Neutrophil-Activating Peptide NAP-2 With Enhanced Biological Activity is Truncated at the C-Terminus:
Balcarek Joanna Maria
Bhatnagar Pradip Kumar
King Andrew Garrison
Pelus Louis Martin
Fitzgerald David L.
Hall Linda E.
Kinzig Charles M.
SmithKline Beecham Corporation
Venetianer Stephen A.
LandOfFree
Truncated gro and KC chemokines having enhanced bioactivity does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Truncated gro and KC chemokines having enhanced bioactivity, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Truncated gro and KC chemokines having enhanced bioactivity will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1781882