Date: Thu May 29th 2025
Time: 2pm UK time (other times)
Place: Zoom link
Previous Meeting: issue #23
Who can come?: Anyone. No need to say anything unless you'd like to. If you'd like to contribute, please join Github.
Attendees: Dr Eve Carter (UCL), Dr Madison Edwards (Uni Toronto), Prof Matt Todd (UCL), Prof Al Edwards (SGC), Prof Karl Swann (Uni Cardiff), Dr Claudia Tredup (Goethe-University Frankfurt), Anna Girtle (Uni Toronto).
Decks: Eve and Madison's presentation
Agenda:
In meeting:
Eve went over the progress made to find compounds that bind PLCz1. An affinity selection mass spectrometry (ASMS) screen was performed at SGC Toronto targeting PLCz1, with PLCγ included as a counter-screen. This screen identified 11 hit compounds, which Eve acquired along with related analogs (she refers to them as “series” where each series is a ASMS hit and its analogs). Eve then tested these compounds in the Aldol 518 assay while Madison tested them in SPR assays and Anna at Nuvisan tested them in XY-69, IP-One, LIPR2. Eve highlighted that most analogues across series 1–10 showed poor or inconsistent results across assays, except series 11, which warrants deeper investigation.
Madison explained some of the basics of SPR. She confirmed SPR assays on chicken PLCz1 with the Nuvisan control compound. She noticed a specificity issue, as the control compound also binds an unrelated protein, WDR91, indicating non-specific or “sticky” behaviour. Madison gave another example of compound 1e from the ASMS screen which also shows binding to both PLCz1 and WDR91. This compound initially looked promising but due to these results will be dropped. On the other hand, she highlighted a promising “11 series” compound (11a) which exhibited clean SPR binding to PLCz1 but minimal binding to WDR91, demonstrating specificity. This compound was flagged as high-priority for future work.
Eve emphasized that the most promising hits (11e, 11n, 11o) share a common structural motif of a saturated n-heterocycle with an aromatic group on the adjacent carbon to the nitrogen. She plans to perform some amide-coupling of an acid to different amines available in the lab to synthesize more analogues around this scaffold. Hopefully this will improve upon the weaker results seen thus far. Madison mentioned that she shared these structures with the DEL group to aid modelling. Al suggested testing the most promising hits against the inactive human mutant to check for species-selectivity, but Madison noted the limited availability of tagged human protein.
Karl has tested one of Nuvisan’s compounds in their assay and they do not see inhibition of PLCz1 except at toxic concentrations. There was some inhibition of fertilization, which is suspected to be due to inhibition of sperm-egg fusion, not PLCz1 itself. He suggests that to be viable, compounds should show micromolar potency in assays without toxic off-target or membrane effects. He will present this work in more detail in the next meeting.
To do:
L'esprit de l'escalier
If you'd like to follow up after the meeting, please comment below. You can also email, but please be clear if anything in the email should not be public domain - the default is open.
Next meeting: Thu 3rd July 2025, 3pm London time
Date: Thu May 29th 2025
Time: 2pm UK time (other times)
Place: Zoom link
Previous Meeting: issue #23
Who can come?: Anyone. No need to say anything unless you'd like to. If you'd like to contribute, please join Github.
Attendees: Dr Eve Carter (UCL), Dr Madison Edwards (Uni Toronto), Prof Matt Todd (UCL), Prof Al Edwards (SGC), Prof Karl Swann (Uni Cardiff), Dr Claudia Tredup (Goethe-University Frankfurt), Anna Girtle (Uni Toronto).
Decks: Eve and Madison's presentation
Agenda:
In meeting:
Eve went over the progress made to find compounds that bind PLCz1. An affinity selection mass spectrometry (ASMS) screen was performed at SGC Toronto targeting PLCz1, with PLCγ included as a counter-screen. This screen identified 11 hit compounds, which Eve acquired along with related analogs (she refers to them as “series” where each series is a ASMS hit and its analogs). Eve then tested these compounds in the Aldol 518 assay while Madison tested them in SPR assays and Anna at Nuvisan tested them in XY-69, IP-One, LIPR2. Eve highlighted that most analogues across series 1–10 showed poor or inconsistent results across assays, except series 11, which warrants deeper investigation.
Madison explained some of the basics of SPR. She confirmed SPR assays on chicken PLCz1 with the Nuvisan control compound. She noticed a specificity issue, as the control compound also binds an unrelated protein, WDR91, indicating non-specific or “sticky” behaviour. Madison gave another example of compound 1e from the ASMS screen which also shows binding to both PLCz1 and WDR91. This compound initially looked promising but due to these results will be dropped. On the other hand, she highlighted a promising “11 series” compound (11a) which exhibited clean SPR binding to PLCz1 but minimal binding to WDR91, demonstrating specificity. This compound was flagged as high-priority for future work.
Eve emphasized that the most promising hits (11e, 11n, 11o) share a common structural motif of a saturated n-heterocycle with an aromatic group on the adjacent carbon to the nitrogen. She plans to perform some amide-coupling of an acid to different amines available in the lab to synthesize more analogues around this scaffold. Hopefully this will improve upon the weaker results seen thus far. Madison mentioned that she shared these structures with the DEL group to aid modelling. Al suggested testing the most promising hits against the inactive human mutant to check for species-selectivity, but Madison noted the limited availability of tagged human protein.
Karl has tested one of Nuvisan’s compounds in their assay and they do not see inhibition of PLCz1 except at toxic concentrations. There was some inhibition of fertilization, which is suspected to be due to inhibition of sperm-egg fusion, not PLCz1 itself. He suggests that to be viable, compounds should show micromolar potency in assays without toxic off-target or membrane effects. He will present this work in more detail in the next meeting.
To do:
L'esprit de l'escalier
If you'd like to follow up after the meeting, please comment below. You can also email, but please be clear if anything in the email should not be public domain - the default is open.
Next meeting: Thu 3rd July 2025, 3pm London time