SOLUBLE NON-DEPOT INSULIN CONJUGATES AND USES THEREOF

• US 20120010134A1
• Filed: 01/27/2010
• Published: 01/12/2012
• Est. Priority Date: 03/20/2009
• Status: Active Grant

• Current Assignee: SmartCells, Inc. (Merck & Co., Inc.)
• Original Assignee: SmartCells, Inc. (Merck & Co., Inc.)
• Inventors: Zion, Todd C., Lancaster, Thomas M.
• Granted Patent: US 8,569,231 B2
• Time in Patent Office: Days
• US Class Current: 514/6.2
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 47/6415 Toxins or lectins, e.g. clostridial toxins or Pseudomonas exotoxins
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/4713 Autoimmune diseases, e.g. Insulin-dependent diabetes mellitus, multiple sclerosis, rheumathoid arthritis, systemic lupus erythematosus; Autoantigens
• CPC Class Codes: C07K 14/4743 Insulin-like growth factor binding protein
• CPC Class Codes: C07K 14/62 Insulins
• CPC Class Codes: C07K 14/65 Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2

Abstract

In one aspect, the disclosure provides a conjugate comprising an insulin molecule having an A-chain and a B-chain; an affinity ligand covalently bound to the A-chain; and a monovalent glucose binding agent covalently bound to the B-chain, wherein the affinity ligand competes with glucose for non-covalent binding with the monovalent glucose binding agent. In the absence of glucose, the monovalent glucose binding agent binds the affinity ligand to produce a “closed” inactive form of the insulin molecule. When free glucose is added, it competes with the affinity ligand for binding with the monovalent glucose binding agent to produce an “open” active form of the insulin molecule. The monovalent glucose binding agent and affinity ligand are covalently bound to the insulin molecule. The disclosure also provides methods of using these conjugates and methods of making these conjugates. In another aspect, the disclosure provides exemplary conjugates. The disclosure also provides alternative conjugates that are not necessarily activated by glucose.

Claims

1. A conjugate comprising:

an insulin molecule having an A-chain and a B-chain;

an affinity ligand covalently bound to the A-chain; and

a monovalent glucose binding agent covalently bound to the B-chain, wherein the affinity ligand competes with glucose for non-covalent binding with the monovalent glucose binding agent.

2. The conjugate of claim 1, wherein in the absence of glucose, the monovalent glucose binding agent binds the affinity ligand to produce an inactive form of insulin.

3. The conjugate of claim 1, wherein in the presence of excess glucose, glucose competes with the affinity ligand for binding with the monovalent glucose binding agent to produce an active form of insulin.

27. The conjugate of claim 1, wherein the affinity ligand includes a saccharide selected from the group consisting of glucose, mannose, glucosamine, mannosamine, methylglucose, methylmannose, ethylglucose, and ethylmannose.

28. The conjugate of claim 1, wherein the affinity ligand include a bimmanose or trimannose.

29. The conjugate of claim 1, wherein the affinity ligand includes aminoethylglucose (AEG), aminoethylmannose (AEM), aminoethylbimannose (AEBM), or aminoethyltrimannose (AETM).

33. The conjugate of claim 1, wherein the monovalent glucose binding agent is a monovalent lectin.

39. The conjugate of claim 1, wherein the monovalent glucose binding agent is a peptide aptamer.

40. The conjugate of claim 1, wherein the monovalent glucose binding agent is an antibody or antibody fragment.

41. The conjugate of claim 1, wherein the monovalent glucose binding agent is a polynucleotide aptamer.

47. The conjugate of claim 1, wherein the monovalent glucose binding agent is chemically modified.

48. The conjugate of claim 1, wherein the monovalent glucose binding agent is pegylated.

72. A method comprising administering a conjugate of claim 1 to a patient.

73. The method of claim 72, wherein the patient is diabetic.

82. A pharmaceutical composition comprising a conjugate of claim 1 and a pharmaceutically acceptable carrier.

85. A kit comprising a pharmaceutical composition of claim 82 and a syringe or pen.

87. The kit of claim 85, wherein the pharmaceutical composition is in a container separate from the syringe or pen.

88. The kit of claim 87, wherein the pharmaceutical composition comprises a non-liquid carrier and the kit further comprises a liquid carrier in a container separate from the syringe or pen.

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