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A major cause of the paucity of new starting points for drug discovery is the lack of interaction between academia and industry. Much of the global resource in biology is present in universities, whereas the focus of medicinal chemistry is still largely within industry. Open source drug discovery, with sharing of information, is clearly a first step towards overcoming this gap. But the interface could especially be bridged through a scale-up of open sharing of physical compounds, which would accelerate the finding of new starting points for drug discovery. The Medicines for Malaria Venture Malaria Box is a collection of over 400 compounds representing families of structures identified in phenotypic screens of pharmaceutical and academic libraries against the Plasmodium falciparum malaria parasite. The set has now been distributed to almost 200 research groups globally in the last two years, with the only stipulation that information from the screens is deposited in the public domain. This paper reports for the first time on 236 screens that have been carried out against the Malaria Box and compares these results with 55 assays that were previously published, in a format that allows a meta-analysis of the combined dataset. The combined biochemical and cellular assays presented here suggest mechanisms of action for 135 (34%) of the compounds active in killing multiple life-cycle stages of the malaria parasite, including asexual blood, liver, gametocyte, gametes and insect ookinete stages. In addition, many compounds demonstrated activity against other pathogens, showing hits in assays with 16 protozoa, 7 helminths, 9 bacterial and mycobacterial species, the dengue fever mosquito vector, and the NCI60 human cancer cell line panel of 60 human tumor cell lines. Toxicological, pharmacokinetic and metabolic properties were collected on all the compounds, assisting in the selection of the most promising candidates for murine proof-of-concept experiments and medicinal chemistry programs. The data for all of these assays are presented and analyzed to show how outstanding leads for many indications can be selected. These results reveal the immense potential for translating the dispersed expertise in biological assays involving human pathogens into drug discovery starting points, by providing open access to new families of molecules, and emphasize how a small additional investment made to help acquire and distribute compounds, and sharing the data, can catalyze drug discovery for dozens of different indications. Another lesson is that when multiple screens from different groups are run on the same library, results can be integrated quickly to select the most valuable starting points for subsequent medicinal chemistry efforts., Author Summary Malaria leads to the loss of over 440,000 lives annually; accelerating research to discover new candidate drugs is a priority. Medicines for Malaria Venture (MMV) has distilled over 25,000 compounds that kill malaria parasites in vitro into a group of 400 representative compounds, called the "Malaria Box". These Malaria Box sets were distributed free-of-charge to research laboratories in 30 different countries that work on a wide variety of pathogens. Fifty-five groups compiled >290 assay results for this paper describing the many activities of the Malaria Box compounds. The collective results suggest a potential mechanism of action for over 130 compounds against malaria and illuminate the most promising compounds for further malaria drug development research. Excitingly some of these compounds also showed outstanding activity against other disease agents including fungi, bacteria, other single-cellular parasites, worms, and even human cancer cells. The results have ignited over 30 drug development programs for a variety of diseases. This open access effort was so successful that MMV has begun to distribute another set of compounds with initial activity against a wider range of infectious agents that are of public health concern, called the Pathogen Box, available now to scientific labs all over the world (www.PathogenBox.org).

The validation of a fluorescence-based high-throughput screening (HTS) assay for determining the efficacies of large chemical libraries against Babesia divergens (bovine strain) in in vitro cultures was evaluated in this study. Hematocrits (HCTs) of 2.5%, 5%, and 10% were used for the in vitro culture at 1% parasitemia without daily replacement of the medium. Linearity and HTS assay results revealed that the best HCTs were 5% and 10%. The obtained IC50 values of diminazene aceturate, either by fluorescence-based HTS assay with and without daily replacement of medium or by fluorescence- and microscopy-based methods, did not differ significantly at 5% HCT. Actinonin and chloroquine diphosphate were the most effective drugs against the in vitro growth of B. divergens, followed by pyronaridine tetraphosphate- and luteolin-treated cultures. On contrary, tetracycline hydrochloride and (-)-epigallocatechin-3-gallate from green tea exhibited poor activity as compared with diminazene aceturate (positive control drug). The data indicated that 5% HCT without daily replacement of the culture medium mixed with bovine serum in vitro using a fluorescence-based HTS assay creates the best conditions for large-scale drug screening against B. divergens that infect cattle.

This study aimed to correlate the histopathological changes in mice liver with alterations either in liver tissue antioxidants enzymes (catalase and reduced glutathione (GSH)), oxidative stress marker (malondialdehyde (MDA)) or in serum liver function parameters (Alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein (T.P.) and albumin) to predict the outcome of schistosomiasis. Fourty male Swiss albino mice were used in this study and infected with Schistosoma mansoni for 2, 4, 6 and 8 weeks (8 mice for each group), while, the uninfected mice were used as negative control. Liver tissue antioxidants enzymes, oxidative stress marker and serum liver function parameters were determined in coincide with the liver tissue histopathological changes. All selected biochemical makers showed a strong significant positive correlation (p < 0.05) with liver histopathology score except serum albumin and liver tissue catalase enzyme. The last two parameters exhibited negative correlation with liver histopathology score. These results revealed that the more increase in the level of AST, ALT, T.P. and globulin in serum or liver tissue MDA and GSH indicating severs histopathological changed into the affected liver and hopeless prognosis is expected. On contrary, the increase in albumin level in serum or catalase level in liver tissue of affected patient/animal demonstrating mild liver histopathological changes. Subsequently, good prognosis and response to anti-schistosomal treatment will be predictable. This study opens the way to predict the outcome of schistosomiasis through easy and rapid biochemical test. Therefore, other studies are required to apply such correlation with other biochemical parameters especially that synthesized into the liver.

A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10%) were used without daily replacement of the medium. The results of a high-throughput screening assay revealed that the best HCT was 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites. The IC50 values of diminazene aceturate obtained by fluorescence and microscopy did not differ significantly. Likewise, the IC50 values of luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not differ between the two methods. In conclusion, our fluorescence-based assay uses low HCT and does not require daily replacement of culture medium, making it highly suitable for in vitro large-scale drug screening against Babesia and Theileria parasites that infect cattle and horses.

In vitro evaluation of chemotherapeutic agents against Babesia and Theileria parasites has become routine, and the effectiveness of these chemicals is usually determined by comparing the parasitemia dynamics of untreated and treated parasites. Although microscopy is widely used to calculate parasitemia, several disadvantages are associated with this technique. The present study evaluated a fluorescence-based method using SYBR green I stain (SG I) to screen antibabesial agents in in vitro cultures of Babesia bovis . The linearity between relative fluorescence units (RFU) and parasitemia was found to be well correlated with a 0.9944 goodness-of-fit ( r 2 ) value. Subsequently, 50% inhibitory concentration (IC 50 ) values were calculated for 3 antiprotozoan agents, diminazene aceturate, nimbolide, and gedunin, by this method. For diminazene aceturate and nimbolide, the IC 50 s determined by the fluorescence-based method (408 nM and 8.13 μM, respectively) and microscopy (400.3 nM and 9.4 μM, respectively) were in agreement. Furthermore, the IC 50 of gedunin determined by the fluorescence-based method (19 μM) was similar to the recently described microscopy-based value (21.7 μM) for B. bovis . Additionally, the Z′ factor (0.80 to 0.90), signal-to-noise (S/N) ratio (44.15 to 87.64), coefficient of variation at the maximum signal (%CVmax) (0.50 to 2.85), and coefficient of variation at the minimum signal (%CVmin) (1.23 to 2.21) calculated for the fluorescence method using diminazene aceturate were comparable to those previously determined in malaria research for this assay. These findings suggest that the fluorescence-based method might be useful for antibabesial drug screening and may have potential to be developed into a high-throughput screening (HTS) assay.

A specific Spectrophotometric titration method is described for the micro-determination of some phenothiazine derivatives in strong orthophosphoric acid medium using the π-acceptor 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) as a mild oxidizing titrant. The wavelengths of maximum absorption of the formed phenothiazonium cation radicals are 500, 515, 530, 533 and 650 nm for trifluoperazine dihydrochloride, promethazine hydrochloride, prochlorperazine dimaleate, chlorpromazine hydrochloride and thiethylperazine dihydrochloride, respectively. Quantitative recoveries are reported for pure drugs and their dosage forms. The method is simple and specific for determining phenothiazine derivatives in presence of their induced degraded oxidation products.

An indirect titrimetric method is described for the determination of some N-substituted phenothiazine derivatives. The method involves the use of 1,3-dibromo-5,5-dimethylhydantoin or N-bromosuccinimide as the titrant. A known excess of either reagent is added and, after a specified time, the residual reagent is determined iodimetrically. The proposed method was applied to the analysis of pharmaceutical preparations containing the drugs, and the results obtained compared favourably with those obtained by pharmacopoeial methods.

An indirect titrimetric method is described for the determination of several thioxanthene derivatives, namely, chlorprothixene, thiothixene and methixene hydrochloride. The method involves the use of 1,3-dibromo-5,5-dimethylhydantion, N-bromosuccinimide or N-bromophthalimide as the titrant. A known excess of one of these reagents is added and, after a specified time, the residual reagent is determined iodimetrically. The proposed method was applied to the analysis of pharmaceutical preparations containing these drugs and the results obtained, compared favourably with those obtained by the pharmacopoeial methods.

A simple and sensitive kinetic method has been developed for the determination of ethamsylate (ESL) in its pharmaceutical preparations. The method is based upon oxidation of ESL with 3-methyl-2-benzothiazolinone hydrazone hydrochloride in presence of cerium(IV) ammonium sulfate at room temperature for 20 min. The absorbance of the reaction product is measured at 514 nm. The absorbance-concentration plot was rectilinear over the range of 430 g/mL (r = 0.9999). The lower detection limit was 0.267 g/mL (9.110 106 M) and the lower quantitation limit was 0.808 g/mL. The different experimental parameters affecting the development and stability of the reaction product were studied and optimized. The proposed method was applied to the determination of ESL in formulations, and the results obtained were in good agreement with those obtained using a reference method. The proposed method was also used for the in vitro detection of ESL in spiked human plasma at its therapeutic concentration level.