Pandemic box data analysis

[dmitrij176]

Following recent data upload in #1, I have created a scheme of 46 active compounds from the pandemic box screen. This is a raw version that will be updated.

Pandemic Box Scheme.zip

[MFernflower]

@wwjvdsande Interesting to note that eberconazole looks like one of the compounds @fantasy121 just finished making and that MMV 1634491 looks like paclobutrazol

[MFernflower]

@wwjvdsande The fact clemizole had an effect made me wonder if diphenhydramine also has an effect

homologs to human neurotransmitter receptors have been found in ameba but no such work has been done on fungi

[wwjvdsande]

@MFernflower interesting findings! I'm not a chemists so for me it is sometimes difficult to assess if a molecule looks alike or not. Do you know if there a chemical libraries available on any of these compounds?

[MFernflower]

@wwjvdsande TCI sells the drug in question super cheap - I actually have a bottle of it in a lab i'm renting out space in but for me to send a gram over to the Netherlands would be cost prohibitive!

[MFernflower]

@wwjvdsande https://www.tcichemicals.com/eshop/en/nl/commodity/D0423/

Used the US link instead of the EU link sorry

[Wilson-Lm]

From the 46 compounds that were active, fludarabine and fenbendazole caught our attention. These 2 belongs to the benzimidazoles group; some of the drugs within this group eg mebendazole has been shown to inhibit fungal growth. So, we are quite interested to look further into the benzimidazoles.

Does anyone have any information about available chemical libraries for the benzimidazoles?

We are also looking for chemical libraries available for triazolopyrimidines. (ones that inhibits the DHODH pathway - like Olorofim). Does anyone know?

[MFernflower]

@wwjvdsande was azelastine ever tested against the fungus???

[wwjvdsande]

@MFernflower, if it was not part of the pathogen box, the stasis box or the pandemic box than it was not tested.

[wwjvdsande]

@dmitrij176 we received the molecules you synthesized. We will test them for activity.

[Wilson-Lm]

Dear all: Updates on the pandemic box.

We have tested all the compounds in the Pandemic Response Box. First by testing at 100uM and 25uM. Next, using data from 25uM tests, we calculated the IC50, then then from the IC50 data, we further did MICs on compounds that were active against M. mycetomatis below 10uM. MIC's were performed using 10 different M. mycetomatis isolates. Attached is the data that we've obtained so far.

19 compounds were found to be active against M. mycetomatis under 10uM. and from this 19, we picked 13 to do MICs. The other 5 were azoles + 1 Olorofim that we have already tested prior to the Pandemic Response Box. (in the attached power point file, you'll find the list and more information regarding these 19 compounds)

From the 13 compounds, the ones that were most potent belong to the Benzimidazoles - Fenbendazole and Carbendazim. They both have a MIC50 of 0.5. The Imidazoles were also potent. Luliconazole has a MIC50 of <0.03125 and Eberconazole at 0.5.

Because we saw that both Fenbendazole and Carbendazim had a good MIC50 (0.5), we did an extra test on Mebendazole (also a benzimidazole). Mebendazole however was a let down. The MIC50 was >16. It will be really nice to find out why Mebendazole doesnt work while Fenbendazole and Carbendazim does. Something with the structure I would say?

As of now, MIC50 of the 6 compounds (5 azoles and olorofim) is not included in the current attahced excel sheet. I will include them once I have recalculated them from prior experiments and publications.

Data from the excel sheet that is attached to this message will also be uploaded into the master sheet we have on github.

19 compounds for MIC50.pptx

Pandemic Box MasterSheet.xlsx

[Wilson-Lm]

The master sheet for MycetOS on GitHub is now 'updated' ! :)

[MFernflower]

Dr.Lim - I suspect the reason you got weird results from the cytoskeletal agents is that some appear to penetrate cells better than others - perhaps active transport of Fenbendazole and Carbendazim? @Wilson-Lm

[MFernflower]

@Wilson-Lm Does albendazole show any effects on the fungus?

[Wilson-Lm]

Dr.Lim - I suspect the reason you got weird results from the cytoskeletal agents is that some appear to penetrate cells better than others - perhaps active transport of Fenbendazole and Carbendazim? @Wilson-Lm

The penetration factor is indeed a major factor to consider. With my limited knowledge, I dare not assume that to be the sole factor, but it may well be that. It is however an interesting direction to pursue.

With the data we have obtain so far on compounds that are active against M. mycetomatis , perhaps we can figure out what properties enable drugs to penetrate the fungus based on their structure...

What are you able to derive by looking at the difference between Mebendazole vs Fenebendazole and Carbendazim judging by their structure?

[MFernflower]

@Wilson-Lm We really don't know why some drugs enter M. mycetomatis and why some do not - if i had to guess maybe active transporter? Mebendazole is not listed as a fungus killer unlike the two actives so just by reading package inserts is how I hypothesized this

[wwjvdsande]

@Wilson-Lm @kym834 @mattodd @bendndi

The MMV compounds able to prolong larval survival have the following modes of action according to MMV: • MMV006357 – this compound came up in various screens and cytotox has been an issue. MMV thinks it has been implicated in some of the lipid kinases however there is not anything firm and clear on this one. So the mode of action currently is still a mystery • MMV022478 – this is a classic PI4K template. It is an irresistible compound (based on MALDA’s resistance selection studies) and so is likely to have polypharmacology. It was a hit from GSK and scored high in their promiscuity score (i.e. hits in multiple screens) so will be cautious with it. • MMV675968 does indeed look very much like a DHFR and originally is reported as active against cryptosporidium. It has also been picked up in all the antibacterial screens that were run using the PB. Though not published yet, a few analogs were screened against ESKAPE pathogens and a clear SAR understanding exists for these pathogens so won’t be surprised to see that it has pan activity. • MMV1782387 belongs to same class as febendazole; antifungal activity is reported in a Japanese patent and no information about MoA is available.