Methods for reducing the mitogenicity of lectin compositions

• US 7,687,608 B2
• Filed: 10/19/2006
• Issued: 03/30/2010
• Est. Priority Date: 10/19/2005
• Status: Active Grant

• Current Assignee: SmartCells, Inc. (Merck & Co., Inc.)
• Original Assignee: SmartCells, Inc. (Merck & Co., Inc.)
• Inventors: Cai, Wenyi, Pecukonis, Matthew, Faucette, Ryan, Lancaster, Thomas M., Zion, Todd C.
• Primary Examiner(s): Tate; Christopher R
• Assistant Examiner(s): Mi; Qiuwen
• Application Number:
  US11/583,993
• Publication Number:
  US 20070110811A1
• Time in Patent Office: 1,258 Days
• Field of Search: None
• US Class Current: 530/396
• CPC Class Codes: A61K 38/25 Growth hormone-releasing factor [GH-RF], i.e. somatoliberin
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/6903 the form being semi-solid, e.g. an ointment, a gel, a hydrogel or a solidifying gel
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C08B 30/18 Dextrin , e.g. yellow canari, white dextrin, amylodextrin or maltodextrin; Methods of depolymerisation, e.g. by irradiation or mechanically
• CPC Class Codes: C08B 37/00 Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof cellulose D21; microbiological processes C12P
• CPC Class Codes: C08B 37/003 Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
• CPC Class Codes: C08B 37/0069 Chondroitin-4-sulfate, i.e. chondroitin sulfate A; Dermatan sulfate, i.e. chondroitin sulfate B or beta-heparin; Chondroitin-6-sulfate, i.e. chondroitin sulfate C; Derivatives thereof
• CPC Class Codes: C08B 37/0072 Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
• CPC Class Codes: C08B 37/0084 Guluromannuronans, e.g. alginic acid, i.e. D-mannuronic acid and D-guluronic acid units linked with alternating alpha- and beta-1,4-glycosidic bonds; Derivatives thereof, e.g. alginates
• CPC Class Codes: Y10S 436/827 Lectins

Abstract

Methods for reducing the T-cell mitogenicity of lectin compositions are provided. In one aspect this is achieved by chemically modifying mitogenic lectin compositions under optimized conditions. Additionally or alternatively, the reduction in T-cell mitogenicity is achieved by removing unmodified subunits chemically modified mixtures. Modified lectin compositions with reduced T-cell mitogenicity are also provided as are uses of the inventive compositions.

Claims

1. A water-insoluble gelled matrix comprising:

therapeutic agent-mannose-containing polymer conjugates; and

chemically modified lectins that bind the mannose-containing polymer portion of the conjugates and thereby cross-link the conjugates to form the water insoluble gelled matrix, wherein the matrix releases the conjugates from a surface of the matrix in a glucose-responsive manner and the chemically modified lectins are obtained by a method comprising the steps of;

providing a mitogenic lectin composition that includes a plurality of mitogenic lectins that bind glucose and mannose, wherein the mitogenic lectins are each comprised of one or more lectin subunits;

reacting the mitogenic lectins with a chemical compound to produce a chemically modified lectin composition, wherein the chemically modified lectin composition exhibits reduced T-cell mitogenicity as compared to the mitogenic lectin composition; and

isolating a portion of the chemically modified lectin composition by glucose affinity chromatography, wherein the isolated portion exhibits reduced T-cell mitogenicity as compared to the chemically modified lectin composition.

2. The matrix of claim 1, wherein the glucose affinity chromatography uses a resin that comprises dextran.

3. The matrix of claim 1, wherein the mitogenic lectin composition includes Concanavalin A.

4. The matrix of claim 1, wherein the chemical compound is a pegylation agent.

5. The matrix of claim 4, wherein the pegylation agent is selected from the group consisting of N-hydroxysuccinimide activated poly(ethylene glycol), succinimidyl ester of poly(ethylene glycol) propionic acid, succinimidyl ester of poly(ethylene glycol) propionic acid butanoic acid, and succinimidyl ester of poly(ethylene glycol) alpha-methylbutanoate.

6. The matrix of claim 1, wherein the chemically modified lectins are incapable of precipitating glycogen comprised entirely of glucose units.

7. The matrix of claim 1, wherein the mannose-containing polymer is mannosylated-glycogen.

8. The matrix of claim 1, wherein the therapeutic agent comprises insulin.

9. The matrix of claim 8, wherein the unmodified lectins comprise Concanavalin A.

10. The matrix of claim 1, wherein the mitogenic lectin composition includes a mitogenic lectin that is multimeric and comprised of identical subunits in its native form.

11. The matrix of claim 10, wherein the mitogenic lectin is tetrameric in its native form and the mitogenic lectin composition comprises monomers, dimers, trimers, or tetramers of the mitogenic lectin, or any mixture thereof.