The United States of America As Represented by the Department of Health

The United States of America, As Represented by the Secretary, Department of Health and Human Services

Other

The United States of America As Represented by the Secretary of

The UAB Research Foundation (the University of Alabama at Birmingham)

Henry C Krutzsch

David D Roberts

Lance A Liotta

William G Stetler-Stevenson

Nenghua Guo

Joanne Murphy-Ullrich

Eric Negre

Neng-hua Guo

John M Sipes

Joseph L Bryant

Elliott Schiffmann

John K Inman

Stacey L Schultz-Cherry

Mary Stracke

Christine Krutzsch, Legal Representative

Solange M F Ribeiro

Philip J Browning

Christian Hugo

Marie E Beckner

The present invention relates to a peptide comprising the sequence R&#x2014;X&#x2014;X&#x2014;X&#x2014;X&#x2014;R, wherein Xis selected from the group consisting of N, Q, D and S; Xis selected from the group consisting of V, I and L; Xis selected from the group consisting of R and K; and Xis selected from the group consisting of V, I, L and F; Ris a hydrogen or a peptide of 1 to 6 amino acids, an acyl or an aryl group; and Ris a peptide of 1 to 3 amino acids, a hydroxide or an amide. The invention also relates to partial or full retro-inverso peptides comprising the above sequences The invention also relates to peptide-substrate combination comprising a substrate suitable for cell growth and the peptide of the invention, and to a vascular graft and an artificial blood vessel comprising the peptide-substrate combination. The invention also relates to a pharmaceutical composition and a peptide conjugate comprising the peptide of the invention. The invention also relates to a method of inhibiting adhesion of a cell expressing &#x3b1;3&#x3b2;1 integrin to an extracellular matrix, inhibiting &#x3b1;3&#x3b2;1-integrin-mediated cell motility, inhibiting &#x3b1;3&#x3b2;1-integrin mediated cell proliferation, promoting &#x3b2;3&#x3b2;1-integrin mediated cell proliferation and inhibiting angiogenesis utilizing the peptides of the invention.

Peptides and their utility in modulation of behavior of cells expressing &#x3b1;3&#x3b2;1 integrins

The present invention relates to a peptide comprising the sequence R&#x2014;X&#x2014;X&#x2014;X&#x2014;X&#x2014;R, wherein Xis selected from the group consisting of N, Q, D and S; Xis selected from the group consisting of V, I and L; Xis selected from the group consisting of R and K; and Xis selected from the group consisting of V, I, L and F; Ris a hydrogen or a peptide of 1 to 6 amino acids, and acyl or an aryl group; and Ris a peptide of 1 to 3 amino acids, a hydroxide or an amide. The invention also relates to partial or full retro-inverso peptides comprising the above sequences. The invention also relates to peptide-substrate combination comprising a substrate suitable for cell growth and the peptide of the invention, and to a vascular graft and an artificial blood vessel comprising the peptide-substrate combination. The invention also relates to a pharmaceutical composition and a peptide conjugate comprising the peptide of the invention. The invention also relates to a method of inhibiting adhesion of a cell expressing &#x3b1;&#x3b2;integrin to an extracellular matrix, inhibiting &#x3b1;&#x3b2;-integrin-mediated cell motility, inhibiting &#x3b1;&#x3b2;-integrin mediated cell proliferation, promoting &#x3b1;&#x3b2;-integrin mediated cell proliferation and inhibiting angiogenesis utilizing the peptides of the invention.

Method of diagnosing cancer in a male mammal comprising obtaining and assaying a test sample for an increased level of semenogelin; method of diagnosing cancer in a female mammal comprising obtaining and assaying a test sample for the presence of semenogelin; methods of prognosticating and assessing the effectiveness of treatment of a cancer in a mammal comprising measuring the level of semenogelin in a test sample; method of inducing an immune response to a cancer in a mammal comprising administering to the mammal a composition comprising (a) an immune-response inducing effective amount of (i) semenogelin or (ii) antibody thereto or (b) a recombinant vector encoding and expressing an immune-response inducing effective amount of (i) or (ii); and composition comprising a carrier and (a) an immune-response inducing effective amount of (i) a polypeptide of any of SEQ ID NOS:1-27 or (ii) antibody thereto or (b) a recombinant vector encoding and expressing an immune-response inducing effective amount of (i) or (ii).

Use of semenogelin in the diagnosis, prognosis and treatment of cancer

The present invention relates to a peptide comprising the sequence R-X-X-X-X-R, wherein Xis selected from the group consisting of N, Q, D and S; Xis selected from the group consisting of V, I and L; Xis selected from the group consisting of R and K; and Xis selected from the group consisting of V, I, L and F; Ris a hydrogen or a peptide of 1 to 6 amino acids, and acyl or an aryl group; and Ris a peptide of 1 to 3 amino acids, a hydroxide or an amide. The invention also relates to partial or full retro-inverso peptides comprising the above sequences. The invention also relates to peptide-substrate combination comprising a substrate suitable for cell growth and the peptide of the invention, and to a vascular graft and an artificial blood vessel comprising the peptide-substrate combination. The invention also relates to a pharmaceutical composition and a peptide conjugate comprising the peptide of the invention. The invention also relates to a method of inhibiting adhesion of a cell expressing &#x3b1;3&#x3b2;1 integrin to an cellular matrix, inhibiting &#x3b1;3&#x3b2;1-integrin-mediated cell motility, inhibiting &#x3b1;3&#x3b2;1-integrin mediated cell proliferation, promoting &#x3b1;3&#x3b2;1-integrin mediated cell proliferation and inhibiting angiogenesis utilizing the peptides of the invention.

The present invention provides a method of treating kidney or renal diseases/conditions in a subject by administering to the subject a pharmaceutically effective amount of a purified LAP peptide, a TSP-1 type 1 repeat peptide, or a fragment thereof to interfere with the activation process of TGF-&bgr; by thrombospondin-1 to reduce and/or prevent renal damage. The present invention further provides a method of improving renal function in a subject having impaired renal function by administering to the subject a pharmaceutically effective amount of a purified LAP peptide, a TSP-1 type 1 repeat peptide, or a fragment thereof.

Use of peptides inhibitory for thrombospondin dependent TGF-&bgr; activation in the treatment of kidney disease

The invention provides a method of stimulating TGF-&bgr; activity, comprising contacting latent TGF-&bgr; with an amount of an activating peptide from TSP effective to convert latent TGF-&bgr; to active TGF-&bgr;. Also provided is a method of inhibiting the stimulation of TGF-&bgr; activity, comprising contacting latent TGF-&bgr; with an amount of an inhibiting peptide from TSP effective to inhibit conversion of latent of TGF-&bgr; to active TGF-&bgr;. The invention also provides a method of enhancing wound healing, comprising the step of administering to the wound site an amount of an activating peptide from TSP effective to convert latent TGF-&bgr; to active TGF-&bgr;, the activation of TGF-&bgr; resulting in enhanced wound healing. A method of preventing excessive fibrosis stimulated by TGF-&bgr; in pathology is also provided, comprising administering to a site of potential fibrosis an amount of inhibiting peptide from TSP effective to inhibit conversion of latent TGF-&bgr; to active TGF-&bgr;, resulting in reduced fibrosis. The invention also provides a method of blocking TGF-&bgr;-mediated inhibition of endothelial cell proliferation comprising contacting the endothelial cells with an inhibiting peptide effective to inhibit conversion of latent TGF-&bgr; to active TGF-&bgr;, resulting in proliferation of endothelial cells.

Compositions for stimulating TGF-&bgr; activity

The present invention relates, in general, to AAMP-1, and to a peptide derived from the amino-terminal region of AAMP, P189. In particular, the present invention relates to a DNA segment encoding AAMP-1, P189 or fragments thereof; polypeptides encoded by said DNA segment; recombinant DNA molecules containing the DNA segment; cells containing the recombinant DNA molecule; a method of producing a AAMP-1, and P189 polypeptide or fragments thereof; antibodies specific to AAMP-1; and a method of measuring the amount of AAMP-1 in a sample. The present invention further relates to methods of using AAMP, P189 or fragments thereof in promoting cell-cell or cell-substrate adhesion, wound healing in patients, prosthetic acceptance, concentrating heparin in tissues, and inhibiting metastases and invasion of malignant cells.

Human cell adhesion protein AAMP-1 and uses thereof

This invention identifies a biologically active group of peptide sequences from Type I repeat units of the extracellular matrix protein, human thrombospondin-1, identical or homologous to the sequence, KRFKQDGGWSHWSPWSSC (SEQ ID NO.30). The biological activities residing with the full sequences, portions thereof, and variants of the full or partial sequences are disclosed. The invention describes how biological activity may be enhanced by covalently linking these peptides to suitable carriers, preferably a branched, water-soluble polymer of low (or absent) toxicity and immunogenicity, such as polysucrose (Ficoll.TM.). The invention describes (1) a method for preparing such conjugates, (2) the use of the defined peptides or their conjugates in blocking or modifying the action on cellular processes of heparin (e.g., proliferation, adhesion, motility, extravasation and neovascularization), sulfatides, related sulfated glycoconjugates, fibronectin, and basic fibroblast growth factor, involving malignant cell lines and normal endothelial cells. Use of the defined peptides, analogs or peptidomimetics and their conjugates for treatment of metastatic tumors, breast carcinomas, melanomas, Kaposi's sarcomas, hemangiomas, diabetic retinopathies, and various pathological conditions dependent upon neovascularization is also disclosed.

Heparin and sulfatide binding peptides from the type-I repeats of human

Peptides derived from the second type 1 repeat of human endothelial cell thrombospondin which bind to the gelatin-binding domain of fibronectin have been isolated and synthetically produced. The peptides can be used to bind to fibronectin or other related collagen-binding proteins to inhibit fibronectin-dependent cell adhesion to collagen matrices and to inhibit interactions with collagen of other proteins that share homologies with the gelatin-binding domain of fibronectin.

Peptide inhibitors of fibronectin and related collagen-binding proteins

This invention identifies a biologically active group of peptide sequences from Type I repeat units of the extracellular matrix protein, human thrombospondin-1, identical or homologous to the sequence, KRFKQDGGWSHWSPWSSC (SEQ ID NO:30). The biological activities residing with the full sequences, portions thereof, and variants of the full or partial sequences are disclosed. The invention describes how biological activity may be enhanced by covalently linking these peptides to suitable carriers, preferably a branched, water-soluble polymer of low (or absent) toxicity and immunogenicity, such as polysucrose (Ficoll.TM.). The invention describes (1) a method for preparing such conjugates, (2) the use of the defined peptides or their conjugates in blocking or modifying the action on cellular processes of heparin (e.g., proliferation, adhesion, motility, extravasation and neovascularization), sulfatides, related sulfated glycoconjugates, fibronectin, and basic fibroblast growth factor, involving malignant cell lines and normal endothelial cells. Use of the defined peptides, analogs or peptidomimetics and their conjugates for treatment of metastatic tumors, breast carcinomas, melanomas, Kaposi's sarcomas, hemangiomas, diabetic retinopathies, and various pathological conditions dependent upon neovascularization is also disclosed.

Heparin- and sulfatide binding peptides from the type I repeats of human

Inappropriate degradation of extracellular matrix molecules by metalloproteinases plays an important role in a wide variety of pathologic conditions including neoplasia and arthritis. The present invention is an isolated protein of approximately 23,000 daltons in size which binds to metalloproteinases with high affinity, can be purified using affinity chromatography on solid phase metalloproteinases, and is potentially useful for therapy of pathologic conditions involving the inappropriate production of metalloproteinases. This protein is characterized by the presence of the following amino acid sequences:

Metalloproteinase peptides

The present invention is an isolated protein of 21,600 Da which binds to both latent and activated type IV collagenase with high affinity at 1:1 molar stoichiometry, thereby abolishing enzyme activity. The protein is purified by affinity chromatography on solid phase metalloproteinase, or solid phase metalloproteinase substrates which bind the enzyme-inhibitor complex. The complete primary structure of this protein (initially called CSC-21K), as determined by sequencing overlapping peptides spanning the entire protein, reveals homology with a protein called TIMP, Tissue Inhibitor of Metalloproteinases. In addition, a cDNA for this novel inhibitor, now designated TIMP-2, was cloned from a melanoma cell and its sequence was compared with that of human TIMP-1. Northern blots of melanoma cell mRNA showed two distinct transcripts of 0.9 kb and 3.5 kb which are down-regulated by transforming growth factor-.beta., and are unchanged by phorbol ester treatment. The inhibitor of the present invention may be used for treatment of pathologic conditions resulting from inappropriate degradation of extracellular matrix molecules by matrix metalloproteinases, such as metastatic neoplasia, myocardial infarction, and arthritis. Therapeutic treatments using this inhibitor may include formulations for inhalation and inclusion complexes adapted for buccal or sublingual administration, or administration of a recombinant DNA molecule which expresses a DNA segment that encodes the matrix metalloproteinase inhibitor of this invention.

Matrix metalloproteinase inhibitor peptides

The present invention relates, in general, to autotaxin. In particular, the present invention relates to a DNA segment encoding autotaxin; recombinant DNA molecules containing the DNA segment; cells containing the recombinant DNA molecule; a method of producing autotaxin; and antibodies to autotaxin.

Autotaxin: motility stimulating protein useful in cancer diagnosis

A family of metalloproteinases exist which cleave extracellular matrix molecules. These metalloproteinases are secreted in a latent inactive form and require activation in order to specifically cleave the preferred substrate. A series of peptides have been prepared based on the complete sequence analysis of type IV procollagenase. Peptide inhibitors were synthesized which correspond to cysteine repeat regions and histidine containing regions; the mechanism of action of these peptides involves inhibition of binding of the enzyme to the substrate. Peptide inhibitors were synthesized which correspond to the peptide cleaved off during activation, and constitute a novel class of metalloproteinase inhibitors. These inhibitors are members of a series of peptides which contain the core amino acid sequence RKPRC or analogs thereof. The cysteine residue is required for activity. Affinity purified antibodies directed against specific peptides can be used to a) detect any general metalloproteinase enzyme with the sequence in part VAAHE or PRCGNPD, and distinguish it from other known members of the metalloproteinase family, b) block functional domains resulting in the inhibition of enzyme activity, and c) distinguish latent from activated forms of the enzyme.

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Bethesda, MD, United States of America

Henry C Krutzsch