UCL triaged the hits from the original screen (see the Origins page), data shown here Anwar Hossain repurchased and stocked them (data on the 7) and shipped aliquots to TAMU (Inna Krieger) for re-testing to confirm their activities.
Another aliquot of the seven re-purchased hits was sent from UNC to Nick Heaton (Duke University), received Nov 15th 2023.
Results from the testing of the seven re-purchased hits at TAMU can be found here.
Results from the testing of the same seven compounds at Duke (using a fluorescence-based assay) showed that two of the seven compounds showed promising activity, data shown here.
The results are in alignment, with the most promising compounds being identified.
Both labs conducted a protein-free control experiment (using compound+probe, no Nsp15, TAMU, data shown here; using probe+Nsp15, add compound after 1h, Duke, data shown here) to ensure that they are not false positives via assay interference.
There are new testing going on at TAMU and new hits will be released once confirmed (when?)
Solubility data
The solubility of the initial seven re-purchased hits was measured by Analiza, Inc.. The two hits with best inhibition results have low solubility. The solubility data was included in this Excelsheet.
Confirmed Hits Expansion
21 analogues were developed based on the structure of the two most promising compounds. Data can be found here
16 Commercially available analogues and starting materials for synthesizing the other 5 analogues were ordered. 5 Compounds were synthesised and shipped to TAMU together with 16 commercial compounds.
Testing results of the 21 analogues from TAMU can be found here.
2nd Hit expansion is ongoing and its details can be found here.
PAINS
However, TAMU employed a bleaching assay and found out the most potent molecule RA-0025381, along with some other molecules are assay interfering, that they can reduce fluorescence reading after Nsp15 become inactive. Their slides can be found here.
Surprisingly, compound RA-0011688 (SACC-0116360) was found assaying interfering in TAMU's FRET assay but not in Duke's FRET assay. Because the probe they used are different, an aliquote of RA-0025381 will be shipped from TAMU to Duke for further testing.
Hit triage, selection, repurchase and retesting
UCL triaged the hits from the original screen (see the Origins page), data shown here Anwar Hossain repurchased and stocked them (data on the 7) and shipped aliquots to TAMU (Inna Krieger) for re-testing to confirm their activities.
Another aliquot of the seven re-purchased hits was sent from UNC to Nick Heaton (Duke University), received Nov 15th 2023.
Results from the testing of the seven re-purchased hits at TAMU can be found here.
Results from the testing of the same seven compounds at Duke (using a fluorescence-based assay) showed that two of the seven compounds showed promising activity, data shown here.
The results are in alignment, with the most promising compounds being identified.
Both labs conducted a protein-free control experiment (using compound+probe, no Nsp15, TAMU, data shown here; using probe+Nsp15, add compound after 1h, Duke, data shown here) to ensure that they are not false positives via assay interference.
There are new testing going on at TAMU and new hits will be released once confirmed (when?)
Solubility data
The solubility of the initial seven re-purchased hits was measured by Analiza, Inc.. The two hits with best inhibition results have low solubility. The solubility data was included in this Excelsheet.
Confirmed Hits Expansion
21 analogues were developed based on the structure of the two most promising compounds. Data can be found here
16 Commercially available analogues and starting materials for synthesizing the other 5 analogues were ordered. 5 Compounds were synthesised and shipped to TAMU together with 16 commercial compounds.
Testing results of the 21 analogues from TAMU can be found here.
Initial SAR study can be found here.
2nd Hit expansion is ongoing and its details can be found here.
PAINS
However, TAMU employed a bleaching assay and found out the most potent molecule RA-0025381, along with some other molecules are assay interfering, that they can reduce fluorescence reading after Nsp15 become inactive. Their slides can be found here.
Surprisingly, compound RA-0011688 (SACC-0116360) was found assaying interfering in TAMU's FRET assay but not in Duke's FRET assay. Because the probe they used are different, an aliquote of RA-0025381 will be shipped from TAMU to Duke for further testing.