A test system for leukotriene synthesis inhibitors based on the in-vitro differentiation of the human leukemic cell lines HL-60 and Mono Mac 6.
Differentiation of HL-60 cells along the granulocytic lineage by DMSO in the presence of transforming growth factor-beta and low concentrations of 1,25-dihydroxyvitamin D3 leads to the upregulation of 5-lipoxygenase activity in 100,000 g supernatants and intact cells to levels which are comparable to normal granulocytes. Similarly, differentiation of the human monocytic cell line Mono Mac 6 by 1,25-dihydroxyvitamin D3 and transforming growth factor-beta strongly upregulates the 5-lipoxygenase pathway. Here, we describe an assay system for leukotriene biosynthesis inhibitors which is based on the in-vitro differentiation of HL-60 and Mono Mac 6 cells. Different leukotriene biosynthesis inhibitors like the nonredox type inhibitor ZM 230487, the redox type inhibitor BW A4C and the FLAP inhibitor MK886 were tested and the results were compared with an assay system based on normal human granulocytes. ZM 230487, BWA4C and MK886 showed similar potencies in these cell lines as compared to normal leukocytes. Thus, the in-vitro differentiation of HL-60 and Mono Mac 6 cells provides an excellent model for the screening of drugs affecting the 5-lipoxygenase pathway.
Werz
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Institute of Pharmaceutical Chemistry, University of Frankfurt, Germany.
Schneider
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N
Brungs
M
M
Sailer
E R
ER
Safayhi
H
H
Ammon
H P
HP
Steinhilber
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D
D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D044482:Benzeneacetamides; D002454:Cell Differentiation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D004790:Enzyme Induction / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D007951:Leukemia, Myeloid / Q000201:enzymology / Q000378:metabolism; D020024:Leukotriene Antagonists; D015289:Leukotrienes / Q000096:biosynthesis; D011714:Pyrans / Q000494:pharmacology; D015363:Quinolones / Q000494:pharmacology; D014315:Triterpenes / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000201:enzymology / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects; D002722:Chlorobenzenes
D001094:Arachidonate 5-Lipoxygenase; D044482:Benzeneacetamides; D006877:Hydroxamic Acids; D007211:Indoles; D020024:Leukotriene Antagonists; D015289:Leukotrienes; D011714:Pyrans; D015363:Quinolones; D014315:Triterpenes; C094432:acetyl-11-ketoboswellic acid; C060893:MK-886; D002722:Chlorobenzenes; C056614:N-(3-phenoxycinnamyl)acetohydroxamic acid; C088450:ZM 230487