Biosynthetic Pathway of UDP-N-Acetylglucosamine (UDP-GlcNAc)

UDP-GlcNAc is an important substrate in the intracellular glycosylation reaction process. It participates in the synthesis of sialic acid, mannose, lactic acid, and other glycosyl structures, and serves as a substrate for sugar chain transfer reactions to provide sugar supply. The sugar nucleotide precursor of GlcNAc is UDP-GlcNAc, which undergoes a multi-step enzyme cascade to promote synthesis. There are four consecutive enzymatic reactions occur in eukaryotic cells: (a) Glucose is converted to glucosamine-6-phosphate (GlcN-6P); (b) GlcN-6P is acetylated to GlcNAc-6P; (c) GlcNAc-6P is isomerized to GlcNAc-1P; (d) GlcNAc-1P is uridylated to UDP-GlcNAc. In prokaryotic cells, the difference is that GlcN-6P is first isomerized to GlcN-1P and then N-acetylated. Each reaction step is catalyzed by a separate enzyme, including GlcN-6P synthase, GlcN-6P acetyltransferase, phosphate acetylglucosamine mutase, and UDP-GlcNAc pyrophosphorylase (UAP1), which are considered potential drug targets.

Fig.1 Biosynthetic pathway of UDP-GlcNAc. (Raimi, et al., 2020)

Various UDP-GlcNAc Metabolic Enzyme Inhibitor Development Services at CD BioGlyco

Based on the biosynthetic pathway of UDP-GlcNAc, CD BioGlyco provides different types of metabolic enzyme inhibitor development services.

• GlcN-6P synthase

Many GlcN-6P synthase inhibitors of natural or synthetic origin exhibit bactericidal properties. Synthetic compounds include amino acid anti-capsaicin and oligopeptides. The mechanism of action of oligopeptide antifungals containing GlcN-6P synthase inhibitors involves the transport of the oligopeptide permease, which is cleaved by intracellular peptidases, and the enzyme inhibitor thus released inactivates the GlcN-6P synthase.

• UAP1

The phosphate group of GlcNAc-1P attacks the UTP α,β-phosphodiester bond, thereby generating two products: UDP-GlcNAc and PPi. This forward reaction is inhibited if the α,β-methylene diphosphonate analog of UTP (meUTP), in which the easily broken α,β-pyrophosphate bond is replaced by methylene diphosphonate. CD BioGlyco synthesizes meUTP through conventional phosphorylation, which may inhibit UAP1.

In addition, CD BioGlyco uses high-throughput screening technology to screen large-scale compound libraries for compounds that interact with enzyme targets involved in the UDP-GlcNAc synthesis pathway. Meanwhile, we also provide inhibitor activity assessment services, including in vitro experiments (such as enzyme activity assays) and in vivo experiments (such as cell or animal models) to verify the activity, toxicity, and pharmacokinetic properties of the screened compounds.

Fig.2 Various UDP-GlcNAc metabolic enzyme inhibitor development services. (CD BioGlyco)

Publication

Technology: Crystal structure determination technology, Ion exchange chromatography.

Journal: RSC Chemical Biology

IF: 4.1

Published: 2020

Results: The authors reported the crystal structure of A. fumigatus UAP1 (AfUAP1) in a complex with two reaction products, UDP-GlcNAc and pyrophosphate. AfUAP1 has a three-domain fold with a central catalytic domain. Flanked by two domains of unknown function, AfUAP1 has a three-domain fold, a large active site cleft, whose wall is formed by the central domain and the C-terminal domain, with the pyrophosphorylase domain in gray. The authors revealed key conformational changes of AfUAP1 and the important participation of magnesium ions and Lys437 in catalysis. Inspired by the UDP-GlcNAc mechanism, the authors synthesized a meUTP and used ion exchange chromatography to purify and analyze the product, which provided a basis for the future design of small molecule inhibitors of this type of enzyme.

Fig.3 AfUAP1 conformation. (Raimi, et al., 2020)

Applications

• Antimicrobial drug research: UDP-GlcNAc plays a key role in bacterial and fungal cell wall synthesis. The development of UDP-GlcNAc inhibitors is applied in research into antibacterial and antifungal treatments to interfere with bacterial and fungal cell wall synthesis.
• Anti-tumor research: UDP-GlcNAc is involved in the abnormal glycosylation process of tumor cells. By developing UDP-GlcNAc inhibitors, it is possible to block glycosylation of tumor cells and study their potential applications in anti-tumor therapy.
• Regulation of immune cell activation: UDP-GlcNAc participates in the activation process of immune cells. By developing UDP-GlcNAc inhibitors, the activation status of immune cells is regulated, thereby studying diseases related to immune diseases, such as rheumatoid arthritis, autoimmune diseases, etc.

Advantages

• CD BioGlyco has an experienced team of scientists and professionals who design and optimize suitable compounds to inhibit the activity of the UDP-GlcNAc enzyme.
• We will develop personalized strategies for UDP-GlcNAc inhibitor development based on our client's needs and goals, including compound screening, molecular optimization, activity assessment, etc. Customized strategies help ensure the best R&D solutions to achieve clients' specific goals.
• We provide various advanced technical means to conduct comprehensive performance evaluations of the developed UDP-GlcNAc inhibitors. Evaluation determines the potency, selectivity, stability, and toxicity properties of inhibitors, providing important information for further development.

CD BioGlyco has rich experience in the field of Glycosylation Inhibitor Development, which provides mature and professional UDP-GlcNAc inhibitor development services. Please feel free to contact us in time if you need detailed service content.