Modified Protein Variants Enhancing GLP-1 Inducibility and Stability
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Solution Overview
Problem
Current GLP-1 inducing proteins, such as B2UM07, face limitations in inducibility, thermal stability, and enzyme resistance, which hinder their effectiveness in reducing body weight and treating metabolic diseases.
Innovation Solution
Development of protein variants with specific amino acid substitutions and modifications, such as truncating the N-terminus and adding a hexa-histidine tag, to enhance GLP-1 inducibility, thermal stability, and enzyme resistance, including variants like those described in SEQ ID NO: 2 and SEQ ID NO: 4, which are engineered for improved performance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the B2UM07 protein is used to induce GLP-1 secretion, then appetite-regulating and metabolic disease treatment effects are achieved, but the protein exhibits insufficient thermal stability and enzyme resistance
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid sequences of the B2UM07 protein through site-directed mutagenesis. Specific amino acid substitutions (e.g., E46A, E46D, E46Q, E46N) were introduced to alter the protein's physical and chemical properties, resulting in variants with enhanced thermal stability and enzyme resistance while preserving or improving GLP-1 inducibility.
Solution Approach 2:
The patent applies local quality by making targeted modifications at specific positions within the protein sequence rather than uniform changes. The mutations are localized to particular regions (e.g., position 46, 59, 60, 82, 94, 95, 96) to optimize specific properties such as thermal stability and enzyme resistance while maintaining the overall protein structure and function.
2Stability of the object's composition
If protein variants with improved thermal stability are developed, then the protein maintains activity at higher temperatures, but the complexity of protein engineering and characterization increases
Solution Approach 1:
The patent applies segmentation by dividing the protein engineering process into systematic stages: identifying target positions for mutation, performing site-directed mutagenesis at specific sites, expressing the mutated proteins in E. coli, and characterizing their properties. This segmented approach manages the complexity of protein engineering through organized, stepwise procedures.
Solution Approach 2:
The patent applies feedback by systematically characterizing each protein variant's properties (thermal stability, enzyme resistance, GLP-1 inducibility) and using this information to guide further optimization. The characterization data feeds back into the design of subsequent mutations, creating an iterative improvement cycle that manages engineering complexity through informed decision-making.
Data Source
AI summary
This disclosure relates to a protein variant having improved GLP-1 inducibility and/or thermal stability and enzyme resistance, and a pharmaceutical composition for reducing body weight and/or body fat, suppressing appetite or treating or preventing of metabolic diseases, comprising the same.