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8th January 2012 @ 10:44
Performed by James Pham and Stuart Ralph, University of Melbourne. Posted here on their behalf. Data posted in full below, but were first discussed [url=https://plus.google.com/114959083191278443851/posts/V8E5z7zz6qA]here[/url] with graphical description of data [url=http://twitpic.com/7sxzgn]here[/url]. [url=https://docs.google.com/spreadsheet/ccc?key=0AjjLI6V9UglkdFdwU2lFSFhzb05YcWZSVkNzOU00NkE&hl=en_US#gid=0]Data for compounds 1-8[/url] (also [url=http://bit.ly/vSGHjB]short link[/url]) [url=https://docs.google.com/spreadsheet/ccc?key=0AjjLI6V9UglkdG15VGxMUnQwUnhUd2FzcXp2dEtPYVE&hl=en_US#gid=2]Data for compounds 9-16[/url] (also [url=http://bit.ly/xsDirV]short link[/url]) [url=https://docs.google.com/spreadsheet/ccc?key=0AjjLI6V9UglkdGR3eWs5d0VPTEE5eEVOcFF4dm1LQ1E#gid=6]Repeat analysis of compounds 1-9[/url] [data]1034[/data] [data]1036[/data] Experimental description from James Pham: For the two known TCMDC compounds - [url=http://malaria.ourexperiment.org/tcmdc_ap/542/Synthesis_of_TCMDC123812_via_acid_chloride_PMY_102.html]PMY10-2[/url] & [url=http://malaria.ourexperiment.org/tcmdc_ap/541/Synthesis_of_TCMDC123794_via_acid_chloride_PMY_112.html]PMY11-2[/url] - the XC50 were 331.14 nM and 53.87 nM respectively (source: ChEMBL db) where the original authors 'denominated this parameter XC50 to indicate that it is an estimation of the usual IC50 values.' They had used a colorimetric assay using LDH activity as a measure of growth as it is very amenable to high throughput screening. For our first trial, the two compounds inhibition profiles were quite similar with both IC50 estimated around 500 nM, but this is hard to say with any certainty, reason being at 1 uM the curve may not have reached a plateau. And other issues of variability as mentioned in my earlier emails. The screening of these compounds was performed using a malaria SYBR green I- based fluorometric assay. The assay uses SYBR green I, which fluoresces when bound to DNA and is used as an index of growth for Plasmodium falciparum parasites in whole erythrocytes. Asynchronous parasites (standard laboratory strain 3D7) were prepared in 96-well plates for a twofold-dilution of the compounds (highest concentration = 1000 nM). Chloroquine was used as a positive control, negative control was vehicle alone (1% DMSO) and a lane of red blood cells served for subtraction of background. Parasites were grown for 48 hours (Gamo et al paper did 72hr incubation to include possible delay death effects).[data]1034[/data]
Attached Files
Depiction of First Results (image)
First Results (chemdraw)