Biological Evaluation of Compounds

Frozen plasma samples collected from in vivo plasmodium berghei were assayed for quantity of PMY 10-6 at 1, 4 and 24 hours.

Time, h	concentration, ng/mL
1	583
4	44.9
24	<5.00

PMY 10-6 has a 3D7 IC50 of 0.4-0.8 μM, therefore ineffective plasma concentrations are reached in under 4 hours.

Data:

See also:
In vivo Plasmodium Berghei mouse trial of initial leads
Human and Mouse Plasma Stability of OSM-S-5/PMY 10-6
Synthesis of TCMDC-123812 via acid chloride (PMY 10-6)

InChi:
InChI=1S/C15H15FN2O3/c1-9-7-13(15(20)21-8-14(17)19)10(2)18(9)12-5-3-11(16)4-6-12/h3-7H,8H2,1-2H3,(H2,17,19)

Data acquired by Sue Charman's Lab, Centre for Drug Candidate Optimisation, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus), 381 Royal Parade, Parkville, Victoria 3052, Australia [edit: 07 May 2015]

Assessed the stability of OSM-S-5 in human and mouse plasma. The compound was stable in human plasma but susceptible to hydrolysis in mouse plasma. Esterase activity is known to be higher in rodents than in other species which was confirmed using a control compound in this assay (p-nitrophenol acetate) so the results are not too surprising.

Mouse plasma: Half-life: 114 minutes, 25% remaining after 240 minutes Human plasma: No measurable loss after 240 minutes.

Data:

A p.o. P.Berghei mouse study at 50 mg/kg revealed no efficacy of OSM-S-35 (ZYH 3-1),OSM-S-5 (TCMDC-123812) or OSM-S-6 (TCMDC-123794) relative to control. Experiments carried out at Swiss TPH. Experiment start 23/04/12.



Method Overview:
"This protocol is performed in the group of Dr Sergio Wittlin at Swiss TPH (Unit of Prof. Reto Brun), for assessing compound efficacy against the Plasmodium berghei GFP ANKA strain in-vivo. Mice are infected intravenously with parasitized red blood cells on day 0 (2 x 107 parasitized erythrocytes per ml). Experimental mice are generally treated at 4, 24, 48, and 72 hours post-infection with an oral dose of the compound (4-day test by Peters) and are compared to an infected control group for reduction in parasitaemia on day 4 (96 hours post-infection) in % and for mean survival (monitored up to 30 days post-infection). A compound is considered curative, if the animal survives to day 30 after infection with no detectable parasites. Other delivery route (intravenous, intraperitoneal, subcutaneous) and dosing regimen (e.g. single dose) are possible.

Percent activities below 40% are regarded as inactive.

To put the data into context, a new lead optimization project should typically show an oral ED50 <50mg/kg while a development candidate will typically have an oral ED90 <10mg/kg"

The raw data in excel format:

Biological testing of the 2nd round of compounds reported on 05/05/12 by James S. Pham from the University of Melbourne.

Visual summary of results and compound structures:


Methods Overview:
"Compounds were tested for inhibition of Plasmodium falciparum growth using a SYBR green I fluorescence based assay. Dose response curves were generated comprising of 8 points using a 2-fold serial dilution for the maximum final concentration of 1000 nM. The lowest concentration tested was 7.81 nM."

Raw Data originally posted on Google+ along with a detailed method description and report.

Raw data and graphs in excel format:


Compounds tested:
PMY 12-5, PMY 27-2, PMY 31-5, PMY 34-1, ZYH 3-1, ZYH 5-1, ZYH 6-1/6-2, ZYH 7-2, ZYH 10-2 A, ZYH 10-2 B, ZYH 12-1/12-2, ZYH 15-1, ZYH 16-1, ZYH 17-1, ZYH 18-1, ZYH 19-1, ZYH 22-3, ZYH 23-1

Analysis of the same compounds:
Second Round of Evaluation by GSK Tres Cantos
Second Round of Evaluation by the Avery Lab

Results largely consistent with above analyses.

Analysis of PMY 2-4, PMY 6-1, PMY 10-2 (TCMDC-123812), PMY 11-2 (TCMDC-123794), PMY 12-1 and PMY 14-1 (acyl near neighbour) by GSK Tres Cantos courtesy of Felix Calderon.

Assay details have been previously reported and are provided in the supporting information.

IC50 48hrs:
PMY 10-2 (TCMDC-123812) 0.818 μM
PMY 11-2 (TCMDC-123794) 0.245 μM
PMY 14-1 0.047 μM
Atovaquone 0.001 μM
Pirimethamine 0.033 μM
Artesunate 0.019 μM
Chloroquine 0.052 μM



See also:
Second Analysis of First Set of Compounds (Avery)
First Set of Biological Data