Glycyrrhetinic Acid Derivative Chemo-Enzymatic Synthesis Service

Glycyrrhetinic Acid (GA) is a triterpenoid with significant medicinal value, but its clinical application is often limited by its low natural activity and the difficulty of efficient derivatization. Traditional chemical modification methods are inefficient, often lack regioselectivity, and yield complex product mixtures.

CD Biosynsis pioneers a solution combining biocatalysis and chemical synthesis. We focus on engineering specific P450 enzymes for difficult C-H bond activation and site-specific hydroxylation, which is nearly impossible to achieve chemically. Our strategy centers on developing high-efficiency 'chemo-enzymatic' synthesis methods to produce novel GA derivatives with enhanced activity and bioavailability. We provide optimized enzymatic systems and streamlined processes to accelerate the R&D and manufacturing of advanced triterpenoid drugs.

Pain Points

Derivatization and production of high-activity Glycyrrhetinic Acid (GA) pose unique challenges:

• Low Activity of Natural GA: The native Glycyrrhetinic Acid molecule often exhibits insufficient therapeutic activity or poor pharmacokinetic properties, necessitating structural modification to enhance its pharmaceutical efficacy.
• Poor Efficiency of Chemical Modification: Traditional chemical synthesis struggles with the triterpenoid structure's complexity, leading to poor regioselectivity and low conversion rates, particularly in activating inert C-H bonds .
• Lack of Site-Specific Modification: It is extremely difficult to chemically introduce functional groups at specific positions (e.g., C-7, C-11) of the GA backbone without affecting other parts of the molecule, limiting the creation of novel derivatives.
• Scale-Up and Waste: Chemical processes for complex derivatives often require multiple steps, leading to high production costs and the generation of large amounts of environmentally hazardous chemical waste.

The solution lies in leveraging the unparalleled precision and efficiency of engineered enzymes to perform site-specific modifications.

Solutions

CD Biosynsis overcomes the chemical synthesis limitations by integrating highly specific biocatalysis into the production pipeline, focusing on precision modification:

Specific P450 Enzyme Engineering for C-H Activation

           

We employ directed evolution and rational design to modify Cytochrome P450 enzymes , giving them the capability to perform highly selective and efficient hydroxylation on inert C-H bonds of the GA structure.

Efficient 'Chemo-Enzymatic' Synthesis Methods

We combine the strength of both worlds: a few efficient chemical steps (e.g., simple esterification) are coupled with the enzymatic precision step (site-specific hydroxylation/methylation) to streamline the overall synthesis route.

Enzyme Expression and Co-Factor Optimization

The engineered P450 system and its necessary NAD(P)H regeneration enzymes are co-expressed in a microbial host (e.g., E. coli or yeast) to ensure high activity and continuous function during the biocatalytic reaction.

Biphasic Reaction System Optimization

To overcome the poor solubility of triterpenoids, we design and optimize biphasic reaction systems (e.g., aqueous/organic phase), enhancing substrate delivery to the enzyme and boosting conversion rate and final yield.

This approach provides precise control over the modification site, leading to new, high-activity GA derivatives with minimal byproduct formation.

Advantages

Choosing CD Biosynsis's GA derivative synthesis service offers the following core value:

Unrivaled Site-Specific Precision

Enzyme catalysis ensures highly specific modification at target C-H bonds , providing pure, single-isomer derivatives that are unattainable via non-selective chemical methods.

Expertise in P450 Engineering

We are specialists in modifying the active sites of P450 enzymes, customizing them to accept the bulky triterpenoid substrate and catalyze the desired transformation efficiently.

High Efficiency and Reduced Steps

The chemo-enzymatic route significantly reduces the number of synthetic steps and eliminates harsh chemical protections/deprotections, boosting overall yield and efficiency.

Access to Novel Active Derivatives

Our method allows for the synthesis of structurally novel derivatives at positions previously inaccessible by chemical means, creating new intellectual property opportunities.

Scalable Biocatalysis System

The engineered enzyme and host are optimized for use in large-scale fermentation or bioreactors , providing a robust and cost-effective solution for industrial-level production.

We are dedicated to accelerating the development of next-generation triterpenoid drugs through innovative biocatalytic methods.

Process

CD Biosynsis's chemo-enzymatic synthesis service follows a standardized research workflow, ensuring every step is precise and controllable:

• Target Definition and Enzyme Sourcing: Define the desired modification site (e.g., C-7, C-11) on GA. Identify and source candidate P450 enzymes or other suitable oxidoreductases.
• Enzyme Engineering and Host Expression: Use directed evolution and high-throughput screening to engineer the enzyme for optimal activity and regioselectivity towards GA. Express the engineered enzyme and necessary co-factor regeneration system in the host strain.
• Biocatalytic Reaction Optimization: Optimize the enzymatic reaction conditions, including pH , temperature, substrate concentration, and the biphasic solvent system , to maximize conversion efficiency and yield.
• Chemo-Enzymatic Protocol Design: Integrate the high-efficiency biocatalytic step into the overall synthetic route, designing the preceding and succeeding simple chemical steps (e.g., final conjugation).
• Result Report Output: Compile a comprehensive report detailing the engineered enzyme sequence, reaction protocol, conversion rate, and NMR/MS-based structural confirmation of the final derivative's purity and regioselectivity.

Technical communication is maintained throughout the process, with timely feedback on enzyme performance and structural validation.

Achieve unparalleled precision in triterpenoid modification! CD Biosynsis provides customized GA derivative synthesis solutions:

• Detailed P450 Engineering and Biocatalysis Protocol , providing guidance for reaction scale-up.
• Contracted clients receive discounts on subsequent bioactivity testing of the synthesized derivatives.
• Experimental reports include complete raw data on conversion rate, yield, and purity , supporting regulatory filing documentation.