Chemical Modification of Insulin Using Flow Chemistry

Area of Science:

• Protein chemistry
• Biopharmaceutical development
• Chemical biology

Background:

• Chemical modification of proteins is crucial for developing molecular probes and biopharmaceuticals.
• Existing protein acylation methods are typically performed in batch mode.
• Lipidation of insulin at LysB29 is used in approved long-acting insulin variants.

Purpose of the Study:

• To report the first use of flow chemistry for the site-selective acylation of insulin.
• To develop and optimize reaction conditions for insulin acylation compatible with microreactor flow chemistry.
• To demonstrate the synthesis of acylated and Boc-protected insulins using this flow chemistry approach.

Main Methods:

• Development of reaction conditions for site-selective acylation at LysB29 using a soluble base.
• Optimization of reagent ratios and flow rates in a microreactor.
• Acylation of insulin with various acids and synthesis of Boc-protected insulins.

Main Results:

• Established flow chemistry conditions for site-selective insulin acylation at LysB29.
• Successfully acylated insulin with a diverse range of acids.
• Synthesized Boc-protected insulins, demonstrating the versatility of the method.
• Insulin remained stable under the developed flow chemistry conditions.

Conclusions:

• Flow chemistry enables site-selective acylation of insulin, a significant advancement in biopharmaceutical modification.
• This method offers a more efficient and potentially scalable approach for producing chemically modified biopharmaceuticals.
• The reduction of byproducts presents a key advantage over traditional batch methods.