Human Insulin Methyl Ester Production via Dioxane Solvent and Buffering

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current methods for producing Human Insulin methyl ester by enzymatic reaction face challenges in reducing impurities like des B (23-29) and Cyclized form of desB-30, and achieving high purity, particularly when using Dimethyl formamide as a solvent.

Innovation Solution

The process employs dioxane as a solvent in combination with sodium carbonate and sodium bicarbonate, which reduces impurities by 40% and 60% respectively, and enhances the purity of Human Insulin Methyl ester by 10% compared to processes using Dimethyl formamide, through specific enzymatic reactions and crystallization conditions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If Dimethyl formamide is used as solvent in enzymatic reaction, then reaction proceeds efficiently, but impurity levels (des B (23-29) and Cyclized form of desB-30) remain high

Engineering Contradiction:
Improvepurity of Human Insulin Methyl esterVSAvoidimpurity levels
Core Design Contradiction:
Manufacturing precisionVSObject-generated harmful factors

Solution Approach 1:

The patent changes the solvent parameter from Dimethyl formamide to Dioxane, and adjusts pH parameters using sodium carbonate and sodium bicarbonate buffers. This parameter change resolves the contradiction by achieving both efficient reaction progression and reduced impurity levels (des B (23-29) reduced by 40%, Cyclized form of desB-30 reduced by 60%), thereby improving manufacturing precision while controlling harmful factors.

Inventive Principle:
Principle #35Parameter changes

2Manufacturing precision

If traditional enzymatic reaction conditions are used, then production process is simple, but purity enhancement is limited

Engineering Contradiction:
Improvepurity of Human Insulin Methyl esterVSAvoidprocess complexity
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent introduces specific parameter changes including pH adjustment using sodium carbonate and sodium bicarbonate buffers, and temperature control during crystallization. These controlled parameter changes achieve 10% purity enhancement without significantly increasing process complexity, as the same enzymatic reaction framework is maintained with optimized conditions rather than fundamentally changing the process architecture.

Inventive Principle:
Principle #35Parameter changes

3Object-generated harmful factors

If dioxane and sodium salts are used in enzymatic reaction, then impurity reduction is achieved, but process conditions become more specific and controlled

Engineering Contradiction:
Improveimpurity levelsVSAvoidflexibility of reaction conditions
Core Design Contradiction:
Object-generated harmful factorsVSAdaptability or versatility

Solution Approach 1:

The patent specifies particular parameter ranges: pH 7.4-8.5 maintained through sodium carbonate and sodium bicarbonate buffers, temperature control during crystallization, and specific solvent composition (dioxane with controlled water content). These controlled parameter changes reduce impurity levels (des B (23-29) by 40%, Cyclized form of desB-30 by 60%) while maintaining reasonable adaptability within defined ranges, allowing the process to be adjusted within established boundaries without requiring complete redesign.

Inventive Principle:
Principle #35Parameter changes

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The use of dioxane and sodium salts in the enzymatic reaction significantly reduces impurities and increases the purity of Human Insulin Methyl ester, achieving higher yields and improved purity profiles compared to traditional methods.

Implementation Method 1

dioxane induces cleavage of insulin precursor by trypsin

Methodology Applied
Scientific EffectEnzymatic hydrolysis: Hydrolysis

Implementation Method 2

enzymatic reaction

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 3

amidation with an L-threonine ester

Methodology Applied
Scientific EffectAmidation reaction: Chemical Bonding

Implementation Method 4

sodium carbonate and sodium bicarbonate

Methodology Applied
Scientific EffectBuffering:

Implementation Method 5

crystallization procedure

Methodology Applied
Scientific EffectCrystallization: Crystallisation

Data Source

PatentEP3087192B1Method of production of human insulin methyl ester
Publication Date: 2020.08.12 BIOCON BIOLOGICS INDIA LTD
  • EP3087192B1 patent drawingFigure 1
  • EP3087192B1 patent drawingFigure 2
  • EP3087192B1 patent drawingFigure 3~4

AI summary

The present invention relates to production of Human Insulin methyl ester by enzymatic reaction. The present invention further relates to production and enhancement of purity of Human Insulin Methyl ester.