Romidepsin Purification pH Control Prevents Adduct Formation
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Solution Overview
Problem
Published methods for preparing romidepsin do not reproducibly yield pure, monomeric romidepsin due to the formation of contaminating dimerized, oligomerized, or polymerized side products, which are difficult to remove and decrease the purity of the active agent.
Innovation Solution
Maintaining an apparent pH less than 6.5, preferably less than 6.0, during purification steps to prevent the formation of these contaminants, using acid or buffer control to monitor and adjust the pH, and employing multiple chromatography steps with resins like SEPABEADS SP850, DIAION HP20SS, and alumina to achieve high purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional purification methods are used, then romidepsin can be isolated from fermentation broth, but dimerized, oligomerized, or polymerized contaminants are formed and purity is reduced
Solution Approach 1:
The patent applies parameter changes by controlling the pH of the purification process to be less than 6.5, preferably less than 6.0. This pH control prevents the formation of dimerized, oligomerized, or polymerized contaminants while maintaining high yield of monomeric romidepsin. The pH parameter is adjusted during chromatography steps to ensure pure product.
2Ease of manufacture
If pH is not controlled during purification, then purification steps can be performed, but contaminants form and reduce product quality
Solution Approach 1:
The patent implements feedback by monitoring and adjusting the pH during purification steps to maintain it below 6.5. This feedback mechanism ensures consistent production of pure monomeric romidepsin and prevents formation of contaminants, making the process reliable and reproducible.
3Manufacturing precision
If multiple chromatography steps are used, then purity can be improved, but process complexity increases
Solution Approach 1:
The patent reduces process complexity by using pH control (parameter change) as the key mechanism to achieve high purity. By maintaining pH below 6.5 throughout the purification process, the patent minimizes the need for multiple complex chromatography steps while still obtaining high purity monomeric romidepsin.
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
This approach results in higher yields and higher purity of monomeric romidepsin, ensuring pharmaceutical-grade quality by minimizing the presence of undesired adducts and extending the shelf-life of romidepsin preparations.
Implementation Method 1
The culture broth is applied to a column packed with SEPABEADS SP850, a non-ionic adsorption resin. Romidepsin is bound to the resin... After washing with city water and 25% aqueous acetone, elution is carried out with about 65% aqueous acetone
Implementation Method 2
The extracted material is purified by batch or column chromatography. Subsequently, multiple column chromatography steps may be used to achieve the desired level of purity
Implementation Method 3
The culture broth is acidified in order to inactivate or kill the microorganisms in the culture
Implementation Method 4
exposure of romidepsin to basic conditions (e.g., greater than an apparent pH of about 7.0) facilitates the reduction of this disulfide bond. Reduced romidepsin with its free thiols has been found to be susceptible to dimerization, oligomerization, or polymerization
Data Source
AI summary
The invention provides an improved process for preparing romidepsin. The process involves producing, purifying, or storing romidepsin under conditions that prevent the formation of undesired adducts. Purifying romidepsin at an apparent pH lower than approximately 6.0 (e.g., between an apparent pH of 4.0 and 6.0) has been discovered to prevent the reduction of the disulfide bond of romidepsin and the subsequent formation of dimerized, oligomerized, or polymerized adducts. The invention also provides compositions of monomeric romidepsin free of dimerized, oligomerized, or polymerized adducts.