The Salk Institute for Biological Studies

The Whittier Institute for Diabetes & Endocrinology

Advanced Targeting Systems, Inc.

Roger Williams General Hospital

Andrew Baird

Douglas A. Lappi

Douglas A Lappi

J Andrew Baird

Andrew J Baird

Ronald G Wiley

Paul Calabresi

Gillian M Beattie

Alberto Hayek

Barbara A Sosnowski

A Raymond Frackelton, Jr

Ward M Casscells

Jeffrey W Clark

Julie G Beitz

Isabel Barthelemy

This invention provides a conjugate comprising Substance P, or an analog thereof, and a protein, such as Saporin, that inhibits protein synthesis. This invention provides a method of reducing the perception of pain by a subject comprising administering to the subject an effective amount of the pharmaceutical composition of the conjugate comprising Substance P, or an analog thereof, and a protein such as Saporin that inhibits protein synthesis, so as to reduce the perception of pain by the subject. This invention provides a method of selectively destroying NK-1R-expressing neuronal cells in a subject comprising administering to the subject an effective amount of the conjugate comprising Substance P, or an analog thereof, and a protein such as Saporin that inhibits protein synthesis, so as to selectively destroy NK-1R-expressing neuronal cells. Lastly, this invention provides a method for treating a NK-1R-associated disorder in a subject, which comprises administering to the subject an amount of the pharmaceutical composition comprising substance P, or an analog thereof, and a protein such as Saporin that inhibits protein synthesis, in an effective amount to treat the disorder associated with the NK-1R.

Substance P-saporin (SP-SAP) conjugates and methods of use thereof

This invention provides a conjugate comprising Substance P, and analogs thereof, and Saporin. This invention provides a method of reducing the perception of pain by a subject comprising administering to the subject an effective dose of the pharmaceutical composition of the conjugate comprising Substance P, and analogs thereof, and Saporin, so as to reduce the perception of pain by the subject. This invention provides a method of selectively destroying NK-1 receptor expressing cells in a subject comprising administering to the subject an effective dose of the conjugate comprising Substance P, and analogs thereof, and Saporin so as to selectively destroy NK-1 receptor expressing cells. Lastly, this invention provides a method for treating a NK-1 receptor associated disorder in a subject, which comprises administering to the subject an amount of the pharmaceutical composition comprising Substance P, and analogs thereof, and Saporin thereby treating the disorder associated with the NK-1 receptor.

Substance P-Saporin (SP-SAP) conjugates and methods of use thereof

Recombinant production of saporin-containing proteins

The invention provides a conjugate comprising FGF or other polypeptide reactive with an FGF receptor, and a cytotoxic agent. The cytotoxic agent can be a ribosome-inactivating protein (RIP), such as saporin, although other cytotoxic agents can also be advantageously used. The cytotoxic agent can be attached to FGF through a chemical bond, or the composition can be prepared as a chimera using techniques of recombinant DNA. The conjugate can be used to treat FGF-mediated pathophysiological conditions by specifically targeting cells having FGF receptors and inhibiting proliferation of or causing death of such cells. Additionally, the conjugate can be used to target cytotoxic agents into cells having FGF receptors to inhibit the proliferation of such cells. The conjugate can be purified on an immobilized-heparin column.

Fibroblast growth factor conjugates

Conjugates comprising bFGF or other FGF polypeptides and a cytotoxic agent are prepared. The cytotoxic agent can be a ribosome-inactivating protein (RIP), such as saporin, which is attached to bFGF through a chemical bond, or the composition can be prepared as a recombinant DNA chimera. The conjugates are used to specifically target cells, in vivo and in vitro, which express FGF receptors. The cytotoxicity of the conjugates is proportional to the number of receptors expressed by a cell type. The conjugate is useful to effectively treat mammals, and in particular human patients, afflicted with tumorigenic conditions, such as human melanomas, human ovarian carcinomas, teratocarcinomas and neuroblastomas, and other FGF-mediated tumors caused by a proliferation of cells which express FGF receptors.

Treatment of tumorigenic pathophysiological conditions with FGF-cytoxic

Patients suffering vascular injury as a result of balloon catheterization or the like are treated with medicaments containing conjugates comprising a ligand, such as bFGF (or another FGF polypeptide), and a cytotoxic agent. The cytotoxic agent can be a ribosome-inactivating protein (RIP), such as saporin, which is attached to the ligand through a chemical bond or prepared as a recombinant DNA chimera. The medicament containing the conjugate is administered IV to patients after they have been treated for atherosclerosis in a manner which commonly results in vascular injury, particularly to the intima, and effectively prevents restenosis. The conjugate kills proliferating smooth muscle cells in the lumen of the blood vessels which surprisingly express large numbers of high-affinity bFGF receptors while not inhibiting the growth of endothelial cells.

Treatment of vascular injury

The invention provides a conjugate comprising FGF or other polypeptide reactive with an FGF receptor, and a cytotoxic agent. The cytotoxic agent can be a ribosome-inactivating protein (RIP), such as saporin, although other cytotoxic agents can also be advantageously used. The cytotoxic agent can be attached to FGF through a chemical bond or the composition can be prepared as a chimera, using techniques of recombinant DNA. The conjugate can be used to treat FGF-mediated pathophysiological conditions by specifically targeting cells having FGF receptors and inhibiting proliferation of or causing death of the cells. Additionally, the conjugate can be used to target cytotoxic agents into cells having FGF receptors, and to inhibit the proliferation of such cells. A method of purifying the conjugate on an immobilized heparin column is also provided.

The addition of basic FGF-saporin mitotoxins selectively elminates fibroblastoids from human islets in culture and increases the ability of such islets to release insulin under basal and stimulated conditions. When such islets are attached to an extracellular matrix, in particular BCEM, the proliferation of islet cells is favored, and the ability of islets cultured in such manner to release insulin is further increased. Moreover, supplementation of the culture media with high glucose or insulin further improves the functioning of the human islets, resulting in augmented insulin release. Combinations of such procedures offer a novel approach towards the establishment of viable human islet cell monolayers for clinical and laboratory research.

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Del Mar, CA, United States of America

Douglas A Lappi