Large Scale Synthesis of N-acetyllactosamine and Human Milk Oligosaccharides

DESCRIPTION provided by applicant In this Phase I project a subset of human milk oligosaccharides HMOs ranging from disaccharides to pentasaccharides will be prepared on scales ranging from several grams to kilograms Specifically N acetyl lactosamine will be chemically prepared in kilogram quantities six HMOs will be chemically prepared on multi gram scales g and several HMOs will be extended on smaller scales g with N acetyl neuraminic acid using a CMP sialic acid donor and commercially available sialyltransferases The primary aims of the project are to chemically prepare key HMO building block molecules on large scales and basic HMO units on modest scales and to demonstrate the commercial practicality of extending several of the latter basic HMOs to larger sialyl containing structures va enzymic modification All syntheses will be conducted under cGLP conditions existing Omicron documents procedures SOPs methods and laboratory policies developed in prior cGMP projects will be modified for use in this work For each oligosaccharide product high resolution H MHz and C MHz NMR data will be obtained to confirm structure and purity The NMR characterization will include an effort to assign the H and C signals as completely as possible with assistance from D homo and heteronuclear datasets For each oligosaccharide product high resolution mass spectral HRMS data will also be obtained to confirm structural assignments made by NMR and high performance liquid chromatography HPLC will be used to confirm chemical purity A major tangible outcome of this project will be the identification and implementation of synthetic methods capable of producing small HMOs on large scales for future pre clinical trials PUBLIC HEALTH RELEVANCE In this Phase I project a subset of human milk oligosaccharides HMOs ranging from disaccharides to pentasaccharides will be prepared in multi gram quantities using concise and reliable methods that are conducive to scale up to support future pre clinical trials The primary aims of the project are to chemically prepare key building block molecules on large kilogram scales to chemically prepare basic HMO units on modest g scales and to demonstrate the commercial practicality of extending the latter basic HMOs to larger sialyl containing structures via enzymic modification on small g scales