Chemical Synthesis of Burkholderia Lipid A Modified with Glycosyl Phosphodiester-Linked 4-Amino-4-deoxy-β-L-arabinose and Its Immunomodulatory Potential

Modification of the Lipid A phosphates by positively charged appendages is a part of the survival strategy of numerous opportunistic Gram‐negative bacteria. The phosphate groups of the cystic fibrosis adapted Burkholderia Lipid A are abundantly esterified by 4‐amino‐4‐deoxy‐β‐L‐arabinose (β‐L‐Ara4N), which imposes resistance to antibiotic treatment and contributes to bacterial virulence. To establish structural features accounting for the unique pro‐inflammatory activity of Burkholderia LPS we have synthesised Lipid A substituted by β‐L‐Ara4N at the anomeric phosphate and its Ara4N‐free counterpart. The double glycosyl phosphodiester was assembled by triazolyl‐tris‐(pyrrolidinyl)phosphonium‐assisted coupling of the β‐L‐Ara4N H‐phosphonate to α‐lactol of β(1→6) diglucosamine, pentaacylated with (R)‐(3)‐acyloxyacyl‐ and Alloc‐protected (R)‐(3)‐hydroxyacyl residues. The intermediate 1,1′‐glycosyl‐H‐phosphonate diester was oxidised in anhydrous conditions to provide, after total deprotection, β‐L‐Ara4N‐substituted Burkholderia Lipid A. The β‐L‐Ara4N modification significantly enhanced the pro‐inflammatory innate immune signaling of otherwise non‐endotoxic Burkholderia Lipid A. Aminosugar‐modified Lipid A: Stereoselective chemical synthesis of Burkholderia Lipid A esterified with β‐L‐Ara4N at anomerically linked phosphate allowed for elucidation of the pro‐inflammatory capacity of the Ara4N modification. The double glycosyl phosphodiester was assembled by coupling of the pentaacylated (4′P)βGlcN(1→6)αGlcN lactol to the anomeric H‐phosphonate of β‐L‐Ara4N (see figure).