Exendin polypeptide linked to biotin, method for the preparation thereof and pharmaceutical composition comprising the same
Inventors
Lee, Kang Choon • Chae, Su Young • Jin, Cheng Hao
Assignees
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Publication Number
US-8466103-B2
Publication Date
2013-06-18
Expiration Date
Abstract
Disclosed are exendin-3 or exendin-4 derivatives modified with biotin, a preparation method thereof and a pharmaceutical composition containing the same. More specifically, disclosed are exendin-3 or exendin-4 derivatives in which the lysine residue of exedin is modified with biotin. The disclosed exendin-3 or exendin-4 derivatives modified with biotin show biological activity similar to that of native exendin and at the same time, have increased in vivo stability and are easily absorbed through the mucosa. Thus, biotin-modified exendin-3 or exendin-4 derivatives are useful for treating diseases, which can be caused by the excessive secretion of insulin, the lowering of plasma glucose, the inhibition of gastric or intestinal motility, the inhibition of gastric or intestinal emptying or the inhibition of food intake. Particularly, the biotin-modified exendin-3 or exendin-4 derivatives are useful for the treatment of diabetes, obesity and irritable bowel syndromes.
Core Innovation
The disclosure provides an exendin polypeptide linked to biotin, in which biotin is bound to at least one of lysine residue 12 and lysine residue 27 of exendin-4 having the amino acid sequence set forth in SEQ ID NO: 2. Position-limited biotin conjugation to defined exendin-4 lysine residues is used to preserve exendin activity while producing a biotin-modified exendin polypeptide.
The biotin-linked exendin polypeptide is intended to increase in vivo half-life and improve mucosal absorption. The biotin-modified exendin polypeptide is associated with improved biological performance, including enhanced stability and maintained functional activity in relevant assays.
The disclosure further describes a method for preparing the biotin-linked exendin polypeptide by reacting exendin-4 (SEQ ID NO: 2) with biotin and a reducing agent, followed by maintaining the reaction mixture under light-shielded conditions, removing unreacted reactants, and separating and purifying the biotin-modified exendin. Analytical characterization and experimental results support enhanced stability and maintained activity, with demonstrated effects on glucose-related parameters and pharmacokinetic behavior.
Claims Coverage
Two independent claims are identified. One claim covers the biotin-linked exendin polypeptide defined by specific biotin-binding lysine residues in exendin-4 (SEQ ID NO: 2), while another claim covers a preparation method for producing the biotin-linked exendin polypeptide using reaction and purification steps under light-shielded conditions. Across the dependent claims associated with these independents, the coverage includes additional binding-site specificity and therapeutic compositions and uses, as well as formulation types and quantitative molar ratio constraints for the method.
Biotin-linked exendin polypeptide with biotin bound to lysine 12 and/or lysine 27
An exendin polypeptide linked to biotin, wherein the biotin is bound to at least one of lysine residue 12 and lysine residue 27 of exendin-4 having the amino acid sequence set forth in SEQ ID NO: 2.
Preparing biotin-linked exendin by reacting exendin-4 with biotin and a reducing agent under light-shielded conditions
A method for preparing an exendin polypeptide linked to biotin, comprising adding exendin-4 (SEQ ID NO: 2), biotin, and a reducing agent to a buffer or an organic solution and allowing the mixture to react; storing the reaction mixture at a given temperature for a given time in a light-shielded condition; removing unreacted reactants; and separating and purifying biotin-modified exendin.
Overall, the claim set covers a specific biotin-conjugated exendin-4 polypeptide where biotin is bound to lysine residue 12 and/or lysine residue 27, and a method to prepare this biotin-modified exendin using reaction with biotin and a reducing agent followed by light-shielded handling, removal of unreacted reactants, and separation and purification. Dependent claims further narrow the biotin binding sites and can add therapeutic compositions with specified indications and dosage form types.
Stated Advantages
Increases in vivo half-life.
Improves mucosal absorption.
Enhanced stability against trypsin and intestinal homogenate.
Maintains insulinotropic activity and GLP-1 receptor binding.
Improves glucose tolerance in diabetic mouse models.
Pharmacokinetic behavior consistent with oral effectiveness (sustained plasma levels/absorption).
Documented Applications
Treating diabetes associated with excessive secretion of insulin.
Treating irritable bowel syndromes, including irritable bowel syndromes associated with lowered plasma glucose and/or inhibition of gastric or intestinal motility, gastric or intestinal emptying, or food intake.
Treating obesity.
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