T. Kawata et al., E5531, a synthetic non-toxic lipid A derivative blocks the immunobiological activities of lipopolysaccharide, BR J PHARM, 127(4), 1999, pp. 853-862
1 The major pathological responses to Gram-negative bacterial sepsis are tr
iggered by endotoxin or lipopolysaccharide. As endotoxin is shed from the b
acterial outer membrane, it induces immunological responses that lead to re
lease of a variety of cytokines and other cellular mediators. As part of a
program aimed at developing a therapeutic agent for septic shock, we have d
eveloped E5531, a novel synthetic lipopolysaccharide antagonist.
2 As measured by release by tumour necrosis factor-alpha, human monocytes o
r whole blood can be activated by lipopolysaccharide, lipid A, and lipoteic
hoic acid (from Gram-positive bacteria). E5531 potently antagonizes activat
ion by all these agents while itself being devoid of agonistic activity.
3 The inhibitory activity of E5531 was dependent on time of addition. When
10 nM E5531 was added simultaneously with lipopolysaccharide or 1-3h before
addition of lipopolysaccharide, production of tumour necrosis factor-alpha
was inhibited by more than 98%. The addition of E5531 1 h after lipopolysa
ccharide reduced the efficacy of E5531 by 47%.
4 Antagonistic activity of E5531 was specific for lipopolysaccharide as it
was ineffective at inhibiting interferon-gamma mediated NO release of RAW 2
64.7 cells, phorbor 12-myristate 13-acetate stimulated superoxide anion pro
duction in human neutrophils, concanavalin A stimulated mitogenic activity
in murine thymocytes and tumor necrosis factor-alpha induced E-selectin exp
ression in human umbilical vein endothelial cells.
5 E5531 as well as MY4, an anti-CD14 antibody, inhibited radiolabelled lipo
polysaccharide binding in human monocytes.
6 These results support our contention that E5531 is a potent antagonist of
lipopolysaccharide-induced release of tumour necrosis factor-alpha and oth
er cellular mediators and may be an effective therapeutic agent for human s
eptic shock due to Gram-negative bacteria.