INO-5042

Screening of fungal and bacterial strains allowed selection of two Streptomyces strains ( S. platensis and S. cinnamonensis) that oxidatively cleave, in moderate to high yields (up to 65% in 24 h), the quinonic ring of a thiazole fused 1,4-naphthoquinone compound, INO5042, used as a model compound for a series of homologous substituted heterocyclic naphthoquinones. The respective products of these whole-cell biotransformations were identified as isomeric phenol-carboxylic acids resulting from a C-C bond cleavage at a position vicinal to each one of the carbonyl groups. The culture and incubation conditions have been optimised and the mechanism of this biotransformation investigated using oxygen isotope incorporation. The results of 18O2 incorporation indicate a dioxygenase reaction, the mechanism of which is discussed in relation with that of hydroquinone-epoxidases, a family of oxygenating enzymes involved in the biosynthesis of polyketide antibiotics in Streptomyces.

Rolling of leukocytes at the surface of the vascular endothelium is a prerequisite for a subsequent firm adhesion, particularly the slow rolling appearing on ELAM CD62E. Therefore, it may be considered that increasing the rolling velocities should be a precise therapeutic target in clinical situations where leukocytes accumulate, mainly venous and arterial ischaemia.

Human neutrophils were allowed to flow on endothelial HUVECs, with and without 4 hours interleukin-1alpha activation, the cells having or not been incubated with INO5042 anti-inflammatory drug. Under a mean shear-stress of 2 dyn/cm(2), rollers and stickers were identified and quantified, using a video-camera and picture analysing software.

When the drug had been added to endothelial cells a shift of velocities was observed towards fast speeds (from 3-5 to 7-11 microm/sec). The same results was significantly found when neutrophils, alone or along with endothelium, had been submitted to the drug, the number of stickers and rollers beeing reduced as well. Finally, such a precise pharmacological method proved efficient to detect the exact mechanism of INO5042 on white cell adhesion.