Claims for Patent: 7,807,405

Summary for Patent: 7,807,405

Title:	Glycoprotein synthesis and remodeling by enzymatic transglycosylation
Abstract:	A chemoenzymatic method for the preparation of a homogeneous glycoprotein or glycopeptide, including (a) providing an acceptor selected from the group consisting of GlcNAc-protein and GlcNAc-peptide; and (b) reacting the acceptor with a donor substrate including an activated oligosaccharide moiety, in the presence of a catalyst comprising endoglycosidase (ENGase), to transfer the oligosaccharide moiety to the acceptor and yield the homogeneous glycoprotein or glycopeptide. The donor substrate includes, in a specific implementation, a synthetic oligosaccharide oxazoline. A related method of glycoprotein or glycopeptide remodeling with a predetermined natural N-glycan or a tailor-made oligosaccharide moiety, and a method of remodeling an antibody including a heterogeneous sugar chain, are also described. The disclosed methodology enables glycoprotein drugs to be modified for prolonged half-life in vivo, reduced immunogenicity, and enhanced in vivo activity, and for targeting and drug delivery.
Inventor(s):	Wang; Lai-Xi (Ellicott City, MD)
Assignee:	University of Maryland Baltimore, Office of Commercial Ventures and Intellectual Property (Baltimore, MD) N/A (N/A)
Application Number:	11/760,434
Patent Claims:	1. A method of generating remodeled and homogeneous functionalized glycopeptide or glycoprotein, the method comprising: (a) preparing an acceptor GlcNAc-containing peptide or protein that accepts an oligosaccharide; and (b) enzymatically adding the oligosaccharide to the acceptor GlcNAc-containing peptide or protein catalyzed by an endoglycosidase enzyme selected from the group consisting of Endo A and Endo M, wherein the oligosaccharide is a synthetic oligosaccharide oxazoline comprising a sugar chain containing monosaccharide residues and glycosidic linkages to form an intermediate glycopeptide or glycoprotein; and (c) adding a biologically active agent or a tag to the intermediate glycopeptide or glycoprotein to form the functionalized glycopeptide or glycoprotein. 2. The method of claim 1, wherein the acceptor GlcNAc-containing peptide or protein is an antibody or fragment thereof. 3. The method of claim 1, wherein the synthetic oligosaccharide oxazoline is a di-, tri-, tetra-, penta-, hexyl-, hepta-, octyl-, nona-, deca-, or undeca-saccharide oxazoline. 4. The method of claim 1, wherein the intermediate glycopeptide or glycoprotein further comprises an azido group; wherein the biologically active agent or tag further comprises an alkynyl group; and wherein in step (c) the adding comprises reacting said azido group and said alkynyl group to form a triazole linker group in said functionalized glycopeptide or glycoprotein. 5. The method of claim 1, wherein the step of preparing an acceptor GlcNAc-containing peptide or protein comprises (a) providing a peptide or protein substrate comprising at least two GlcNAc residues and at least one GlcNAc-GlcNAc bond; and (b) treating the peptide or protein substrate with an endoglycosidase to hydrolyze a bond between two of the GlcNAc residues positioned closest to the peptide or protein substrate thereby forming a peptide or protein substrate with a single GlcNAc-moiety. 6. The method of claim 1, wherein the tag is a drug, toxin, fluorescent probe, biotin, a PEG, lipid, or polypeptide. 7. The method of claim 6, wherein the tag is polypeptide selected from the group consisting of adrenocorticotropic hormone (ACTH); ebiratide; angiotensin; angiotensin II; asparaginase; atrial natriuretic peptide; atrial sodium diuretic peptide; bacitracin; beta-endorphin; blood coagulation factors VII, VIII and IX; blood thymic factor (FTS); blood thymic factor derivatives; bombesin; bone morphogenic factor; bone morphogenic protein (BMP); bradykinin; caerulein; calcitonin gene related polypeptide (CGRP); calcitonins; a cell growth factor selected from the group consisting of EGF, TGF-alpha, TGF-beta, PDGF, acidic FGF, and basic FGF; cerulein; chemokine; cholecystokinin; cholecystokinin-8 (CCK-8); cholecystokinin-pancreozymin (CCK-PZ); colistin; colony-stimulating factor; corticotropin-releasing factor (CRF); cytokine; desmopressin; dinorphin; a dipeptide; dismutase; dynorphin; eledoisin; endorphin; endothelin; endotherin; enkephalin; epidermal growth factor (EGF); erythropoietin (EPO); follicle-stimulating hormone (FSH); gallanin; gastric inhibitory polypeptide; gastrin-releasing polypeptide (GRP); gastrin; G-CSF; glucagon; glutathione peroxidase; glutathio-peroxidase; a gonadotropin selected from the group consisting of human chorionic gonadotrophin (HCG) and .alpha. and .beta. subunits thereof; gramicidin; gramicidines; growth factor (EGF); growth hormone-releasing factor (GRF); growth hormone; human artrial natriuretic polypeptide (h-ANP); human placental lactogen; insulin; insulin-like growth factor; interferon; interleukin; kallikrein; kyotorphin; luliberin; luteinizing hormone (LH); luteinizing hormone-releasing hormone (LH-RH); lysozyme chloride; melanocyte-stimulating hormone (MSH); melanophore stimulating hormone; mellitin; motilin; muramyl; muramyldipeptide; nerve growth factor (NGF); neuropeptide Y; neurotensin; oxytocin; pancreastatin; pancreatic polypeptide; pancreozymin; parathyroid hormone (PTH); pentagastrin; pituitary adenyl cyclase-activating polypeptides (PACAPs); platelet-derived growth factor (PDGF); polymixin B; prolactin; protein synthesis stimulating polypeptide; PTH-related protein; relaxin; renin; secretin; serum thymic factor; somatomedins; thrombopoietin (TPO); thymic humoral factor (THF); thymopoietin; thymosin; thymostimulin; thyroid hormone releasing hormone; thyroid-stimulating hormone (TSH); thyrotropin releasing hormone (TRH); trypsin; tuftsin; tumor growth factor (TGF-alpha); tumor necrosis factor (TNF); tyrocidin; urogastrone; urokinase; vasoactive intestinal polypeptide (VIP); and vasopressin. 8. The method of claim 6, wherein the tag is a drug. 9. The method of claim 2, wherein the GlcNAc containing peptide or protein is an antibody selected from the group consisting of 17b, 48d, A32, C11, 2G12, F240, IgG1b12, 19e, X5, TNX-355 and F91.

Details for Patent 7,807,405

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Microbix Biosystems Inc.	KINLYTIC	urokinase	For Injection	021846	01/16/1978		2039-02-26
Recordati Rare Diseases, Inc.	ELSPAR	asparaginase	For Injection	101063	01/10/1978		2039-02-26
Nps Pharmaceuticals, Inc.	NATPARA	parathyroid hormone	For Injection	125511	01/23/2015		2039-02-26