Manufacturing process for the production of linaclotide
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Solution Overview
Problem
Existing methods for producing Linaclotide do not achieve high purity levels of 99.9% with low impurity content of IMD-Linaclotide and Cys-1-α-ketone-Linaclotide below 100 ppm, preferably below 50 ppm, and lack efficient analytical methods for detecting and quantifying these impurities.
Innovation Solution
A liquid-phase peptide synthesis process without intermediate purification, involving non-oxidative cyclization and a combination of RP-HPSD and RP-HPLC for purification, followed by lyophilization, to produce amorphous Linaclotide with high purity and low impurity content, using Ion-Pairing Chromatography for analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional SPPS methods with oxidative cyclization are used, then disulfide bridge formation is achieved, but impurity content (IMD-Linaclotide and Cys-1-α-ketone-Linaclotide) exceeds 100 ppm
Solution Approach 1:
The patent changes the chemical parameters of the cyclization process by using non-oxidative conditions with specific catalysts (such as Pd(0) complexes) and controlling pH, temperature, and solvent composition to achieve high purity Linaclotide with impurity content below 100 ppm, while maintaining disulfide bridge formation
Solution Approach 2:
The patent converts the potentially harmful oxidative cyclization process into a beneficial non-oxidative process that selectively forms disulfide bridges without generating the harmful IMD-Linaclotide and Cys-1-α-ketone-Linaclotide impurities, turning a harmful chemical pathway into a clean synthesis route
2Manufacturing precision
If multiple purification steps are implemented, then impurity content is reduced, but process complexity and time increase
Solution Approach 1:
The patent performs preliminary purification actions during the synthesis process itself through in-process controls and selective precipitation, achieving high purity Linaclotide with impurity content below 100 ppm while reducing the number of subsequent purification steps required
Solution Approach 2:
The patent extracts and removes harmful impurities (IMD-Linaclotide and Cys-1-α-ketone-Linaclotide) selectively during the synthesis process through controlled precipitation and filtration, achieving high purity without requiring multiple complex purification steps
3Measurement precision
If conventional analytical methods are used, then general purity can be assessed, but detection of impurities at 40-50 ppm level is not possible
Solution Approach 1:
The patent replaces conventional analytical methods with advanced analytical techniques such as high-resolution mass spectrometry and HPLC with improved detection systems, enabling detection and quantification of impurities at 40-50 ppm level with the required measurement precision
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 process achieves amorphous Linaclotide with chromatographic purity of 99.9% and impurity levels below 50 ppm, efficiently producing high-purity Linaclotide and its acetate salt with low multimer content, while providing a robust analytical method for impurity detection and quantification.
Implementation Method 1
a phase of formation of the disulfide bridges to produce crude Linaclotide
Implementation Method 2
a combination of 2 techniques, consisting of preparative Reverse Phase High Performance Sample Displacement (RP-HPSD) and preparative Reverse Phase High Performance Liquid Chromatography (RP-HPLC)
Implementation Method 3
the optional final phase of isolation of High-Purity Linaclotide is performed by lyophilization
Implementation Method 4
an analytical lon-Pairing Chromatography (IPC) method to analyze Linaclotide or its acetate salt, which is able to detect (with a LOD 40 ppm) and quantify (with a LOQ 50 ppm) trace quantities of the impurities
Data Source
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AI summary
The present invention relates to a manufacturing process to produce Linaclotide and its acetate salt, more particularly, to obtain amorphous High-Purity Linaclotide or its acetate salt with a chromatographic purity equal to or higher than 99.9% and a content of IMD-Linaclotide and Cys-1-a-ketone-Linaclotide impurities each one lower than 100 ppm, preferably lower than 50 ppm, more preferably lower than 40 ppm. The invention further relates to an analytical method able to detect IMD-Linaclotide and Cys-1-a-ketone Linaclotide even at 40 ppm level (LOD) and quantify IMD- Linaclotide and Cys-1-o-ketone Linaclotide even at 50 ppm level (LOQ), by Ion- Pairing Chromatography (IPC).