Glycomed, Inc.

Michael Tiemeyer

Brian K Brandley

Margaret Moreland

Stuart J Swiedler

Hans Schweingruber

Paul G James

Narasinga Rao

Robert J Stack

Ligands that bind to human selectin receptors are disclosed. The ligands are formulated with excipient carriers to form compositions which are administered to treated condition such as inflammation. The ligands have the structural formula III ##STR1## or molecules which have hydrogen bond donor groups equivalent to the circled groups with respect to their ability to form hydrogen bonds with a selectin under physiological conditions.

Methods of treating inflammation using selection binding compounds

Ligands in the form of N-acetyllactosamines which bind to endothelial leukocyte adhesion molecule-1 (ELAM-1) are disclosed. The ligand compounds can be formulated into pharmaceutical compositions and/or assay compositions used to alleviate inflammation and assay for the presence of (qualitative) and amount of (quantitative) ELAM-1 and thereby determine the presence, location and degree of inflammation.

Carbohydrate-based anti-inflammatory agents

Ligands in the form of N-acetyllactosamines which bind to endothelial leukocyte adhesion molecule-1 (ELAM-1) are disclosed. The ligand compounds can be formulated into pharmaceutical compositions and/or assay compositions used to alleviate inflammation and assay for the presence of (qualitative) and amount of (quantitative) ELAM-1 and thereby determine the presence, location and degree of inflammation. The ligands are encompassed by general structural formula I as follows: ##STR1## wherein one of F and G is hydrogen and one is an N-acetyl neuraminic acid residue on the terminal unit and are both hydrogen on any other unit; J is hydrogen, a lactosylceramide or a linking group and K is hydrogen or a fucose residue and n is an integer of from 1 to 10 (preferably 3 or 4) with the proviso that n and K are defined such that at least one K is a fucose residue.

Method of treating inflammation by blocking ELAM-1 receptor sites

Method of determining a cite of inflammation utilizing elam-1 ligands

Method of determining a site of inflammation utilizing ELAM-1 ligands

Ligands in the form of N-acetyllactosamines which bind to endothelial leukocyte adhesion molecule-1 (ELAM-1) are disclosed. The ligand compounds can be formulated into pharmaceutical compositions and/or assay compositions used to alleviate inflammation and assay for the presence of (qualitative) and amount of (quantitative) ELAM-1 and thereby determine the presence, location and degree of inflammation. The ligands are encompassed by general structural formula I as follows: ##STR1## wherein one of A and B is hydrogen and one is an N-acetyl neuraminic acid residue; D and E are each independently hydrogen or a fucose residue; n is an integer of from 0 to 10 with the proviso that if n is 0, E is a fucose residue; F is hydrogen, a lactosylceramide residue or a linking group; and molecules equivalent to a compound of formula I regarding its ability to bind to an ELAM-1 receptor to the same degree as a compound of formula I.

Method of determining a site of inflammation utilizing elam-1 ligands

Saccharide in mixtures are separated into groups or individual saccharides and tested for their affinity for particular proteins. The method of the invention is carried out by conjugating saccharides in a mixture with charge-generating moieties and moieties capable of fluorescing such as 4-amino-1-naphthalene sulfonic acid (ANSA) to form conjugates. The saccharide conjugates are subjected to electrophoretic resolution within gels. The resolved bands of material can be seen under ultraviolet light and are electro-blotted onto charged nylon membranes to provide a stable record of the electrophoretic separation. The blots (visible under ultraviolet light) on the nylon membranes are brought into contact with particular proteins in order to determine the binding affinity of these particular proteins to particular saccharides on the nylon membrane. The proteins are preferably bound to labels so that the binding of the proteins to the saccharides can be easily detected.

Electro-blotting of electrophoretically resolved fluorescent-labeled

A method of separating mixtures of saccharides into distinct detectable groups is disclosed. In accordance with the method a tri-functional conjugate must first be provided. The tri-functional conjugate is obtained by reacting a mixture of saccharides with moieties (a) capable of providing a charge upon ionization; (b) capable of fluorescing under ultraviolet light; and (c) having a light-activatable azido group thereon. The different functional moieties may all be present on the same moiety and connected directly to the saccharide or may be connected to each other wherein only one of the moieties is connected directly to the saccharide. The tri-functional conjugates are subjected to electrophoretic separation to obtain separate groups of conjugates in the gel. The groups of conjugates are transferred from the gel to the surface of a membrane which is exposed to light for a sufficient time and light frequency to activate the azido-group. The light-activated azido-groups attached to the surface of the membrane. The tri-functional conjugates can then be contacted with labeled probes such as radiolabeled proteins to determine the affinity of the probes to particular saccharides.

Fluorescent tag for sugar electrophoresis

A method of separating mixtures of saccharides into distinct detectable groups is disclosed. The method comprises modifying 1-amino-4-naphthalene sulfonic acid (ANSA) with a light-sensitive azido-group and binding the modified ANSA to saccharides to form ANSA/saccharide conjugates. The conjugates are subjected to electrophorectic separation to obtain separate groups of conjugates in the gel. The groups of conjugates are transferred from the gel to the surface of a membrane which is exposed to light for a sufficient time and light frequency to activate the azido-group. The light-activated azido-groups attached to the surface of the membrane. The ANSA/saccharides conjugates can then be contacted with labeled probes such as radiolabeled proteins to determine the affinity of the probes to particular saccharides.

Saccharide in mixtures are separated into groups or individual saccharides and tested for their affinity for particular proteins. The method of the invention is carried out by conjugating saccharides in a mixture with 4-amino-1-naphthalene sulfonic acid (ANSA) to form conjugates. The saccharide/ANSA conjugates are subjected to electrophoretic resolution within gels. The resolved bands of material are electro-blotted onto charged nylon membranes to provide a stable record of the electrophoretic separation. The blots on the nylon membranes are brought into contact with particular proteins in order to determine the binding affinity of these particular proteins to particular saccharides on the nylon membrane. The proteins are preferably bound to labels so that the binding of the proteins to the saccharides can be easily detected.

Growing community of inventors

Oakland, CA, United States of America

Michael Tiemeyer