Engineered Transaminases for High-Yield Sitagliptin Synthesis
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Solution Overview
Problem
The current manufacturing process for sitagliptin, a drug used to treat Type 2 diabetes, is inefficient and could be improved to enhance the production of the active pharmaceutical ingredient (API) with higher enantiomeric excess and yield.
Innovation Solution
Engineered transaminases capable of converting the ketoamide substrate to the sitagliptin product with high enantiomeric excess and improved activity, using amino group donors under specific reaction conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If asymmetric hydrogenation using rhodium Josiphos-ligand catalyst is used, then sitagliptin free base is produced with about 97% e.e., but the overall manufacturing yield is limited to 79% from enamine amide substrate
Solution Approach 1:
The patent changes the fundamental reaction parameter from asymmetric hydrogenation to transamination, using engineered transaminase enzymes with optimized amino acid sequences to achieve both high enantiomeric excess and improved yield. The engineered enzymes alter the reaction mechanism and conditions, transforming the limiting factor from catalyst performance to enzyme catalysis efficiency.
Solution Approach 2:
The patent replaces the metal-based catalytic system (rhodium Josiphos-ligand) with a biocatalytic system (engineered transaminase enzymes). This substitution eliminates the need for expensive metal catalysts and complex ligand systems, while achieving superior stereochemical control and yield through protein engineering.
2Manufacturing precision
If multiple processing steps including crystallization upgrade are used, then enantiomeric excess is improved to >99.5%, but the number of steps increases and overall efficiency decreases
Solution Approach 1:
The engineered transaminase enzyme performs the stereochemical transformation in advance with such high enantiomeric excess that subsequent crystallization upgrade steps become unnecessary. The enzyme pre-forms the correct enantiomer in high purity, eliminating the need for additional separation and purification steps.
Solution Approach 2:
The patent extracts and eliminates the crystallization upgrade step from the manufacturing process by achieving sufficient enantiomeric excess directly in the reaction step. This removes an entire processing stage, simplifying the workflow and reducing time and resource requirements.
3Ease of manufacture
If traditional asymmetric hydrogenation process is used, then sitagliptin free base is obtained, but additional reaction steps with phosphoric acid are required to produce the API
Solution Approach 1:
The patent merges the stereochemical transformation and API formation into a single transamination step. The engineered enzyme directly produces the phosphorylated API form rather than requiring separate steps for free base formation and subsequent phosphorylation, consolidating multiple transformations into one operation.
Solution Approach 2:
The engineered transaminase enzyme performs multiple functions: it catalyzes the stereochemical transformation, determines the final API form, and eliminates the need for separate phosphorylation steps. This multi-functional enzyme replaces what previously required multiple specialized reagents and steps.
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 engineered transaminases achieve high conversion rates and enantiomeric excess of the sitagliptin product, enhancing the manufacturing process efficiency and yield.
Implementation Method 1
transaminases capable of converting the ketoamide substrate to the sitagliptin product in presence of an amino group donor
Data Source
AI summary
The present disclosure relates to polypeptides having transaminase activity, polynucleotides encoding the polypeptides, and methods of using the polypeptides.