The Salk Institute for Biological Studies

Selective Genetics, Inc.

The Whittier Institute for Diabetes & Endocrinology

Prizm Pharmaceuticals, Inc.

J Andrew Baird

Douglas A Lappi

Barbara A Sosnowski

Lois Ann Chandler

Nicholas C Ling

Gillian M Beattie

Alberto Hayek

Isabel Barthelemy

Ward M Casscells

Preparations of conjugates of a receptor-binding internalized ligand and a cytocide-encoding agent and compositions containing such preparations are provided. The conjugates contain a polypeptide that is reactive with an FGF receptor, such as bFGF, or another heparin-binding growth factor, cytokine, or growth factor coupled to a nucleic acid binding domain. One or more linkers may be used in the conjugation. The linker is selected to increase the specificity, toxicity, solubility, serum stability, or intracellular availability, and promote nucleic acid condensation of the targeted moiety. The conjugates are complexed with a cytocide-encoding agent, such as DNA encoding saporin. Conjugates of a receptor-binding internalized ligand to a nucleic acid molecule are also provided.

Compositions containing nucleic acids and ligands for therapeutic treatment

Compositions containing nucleic acids and ligands for therapeutic

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

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

Antagonists to basic fibroblast growth factor, a 146 amino acid residue polypeptide, are produced. These antagonists are generally between 10 and 45 residues in length and are characterized by their ability to interact with the FGF receptor and/or inhibit and therefore modulate endothelial and other cell growth in vitro and also in vivo. These antagonists includes the sequence of bovine basic FGF(106-115), namely Tyr-Arg-Ser-Arg-Lys-Tyr-Ser-Ser-Trp-Tyr or a sequence having equivalent residues substituted therein. These peptides are also antagonistic to acidic FGF and other members of the family of FGF peptides. They are effective to combat FGF-promoted mitosis in melanomas and the like.

Fibroblast growth factor antagonists

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.

Maintenance of pancreatic islets

Atrial peptides useful in practising the invention have the formula: ##STR1## wherein R.sub.1 is Arg, Leu-Arg, Ser-Leu-Arg, Arg-Ser-Leu-Arg, Pro-Arg-Ser-Leu-Arg, Gly-Pro-Arg-Ser-Leu-Arg, Ala-Gly-Pro-Arg-Ser-Leu-Arg or Leu-Ala-Gly-Pro-Arg-Ser-Leu-Arg; R.sub.17 is Met or Ile, R.sub.31 is Phe or desR.sub.31 ; R.sub.32 is Arg or desR.sub.32 ; R.sub.33 is Tyr or desR.sub.33 ; and Y is OH or NH.sub.2. These peptides or pharmaceutically acceptable salts thereof, dispersed in a pharmaceutically acceptable liquid or solid carrier, can be administered to mammals, particularly humans, in order to inhibit basal aldosterone secretion and/or to manage aldosterone-dependent hypertention.

Growing community of inventors

San Diego, CA, United States of America

J Andrew Baird