Date: Thu Apr 3th 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), Anna Girtle (Uni Toronto), Dr Jeffrey Mowat (Janssen Pharmaceutica NV), Prof Matilda Katan (UCL), Prof Opher Gileadi (SGC).
Decks: Eve's presentation
Agenda:
In meeting:
In December, SGC Toronto performed ASMS screen against PLCz1 which yielded 11 hits for PLCz1 selective over PLCy1. Eve has since ordered all of the available hits and analogues and has run the Aldol assay with chicken PLCz1. Nuvisan has also used these hits in XY-69 and IP-one assays with human PLCz1, and SGC Toronto is working on performing SPR with chPLCz1. 78 compounds were screened in the Aldol, XY-69, and IP-one assays, with results from each lacking clear correlation. Only a few compounds were found to be active in a single assay, but a few more showed some activity in multiple assays. In particular the “11” series of compounds (see slide deck) seems to have more promising results than the other groups of compounds. Matt suggested looking at the SAR diagrams of these compounds to see if there are any common patterns.
However, Nuvisan also ran LIPR2 assays, which found that the unrelated lipase LIPR2 was inhibited by almost all of the compounds (Note: Eve has not yet discussed this result with Anna, who ran the assay). Madison commented that the fact that LIPR2 is inhibited by so many compounds is biochemically bizarre. She suggests that perhaps there is a contaminant or buffer component that is being carried over in every test that is causing this inhibition. It will be worth looking closer at this.
Madison is performing SPR on these compounds which together with the other assays will help identify any compounds worth further investigation. She has identified a good buffer and has found that the protein is sticking well to the chip and is behaving well. She has tested the Nuvisan compound and say that it has the characteristic “quick on, quick off” binding behavior, making it a good positive control. She has started going through the ASMS hits but the most she has seen so far is sticky binding (slow on, doesn’t come off). This could mean the compound is non-specific, or it could be a specific binder with poor binding.
Madison has also been running DSF experiments to see if there is temperature stabilization, but has not seen any evidence of this yet. She comments that it might not be best assay for our purposes because to her knowledge neither she nor Nuvisan have tested a compound that has shown stabilization in a thermal shift assay. Instead, if there are any strong SPR hits, she will set up crystal trays to try to get the structures.
HitGen have finished the DEL screen of chicken PLCz1 and the data is being analyzed at UNC. Eve and Madison have also started drafting a publication of the PLCz1 results.
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 29 May 2025, 2pm London time
Date: Thu Apr 3th 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), Anna Girtle (Uni Toronto), Dr Jeffrey Mowat (Janssen Pharmaceutica NV), Prof Matilda Katan (UCL), Prof Opher Gileadi (SGC).
Decks: Eve's presentation
Agenda:
In meeting:
In December, SGC Toronto performed ASMS screen against PLCz1 which yielded 11 hits for PLCz1 selective over PLCy1. Eve has since ordered all of the available hits and analogues and has run the Aldol assay with chicken PLCz1. Nuvisan has also used these hits in XY-69 and IP-one assays with human PLCz1, and SGC Toronto is working on performing SPR with chPLCz1. 78 compounds were screened in the Aldol, XY-69, and IP-one assays, with results from each lacking clear correlation. Only a few compounds were found to be active in a single assay, but a few more showed some activity in multiple assays. In particular the “11” series of compounds (see slide deck) seems to have more promising results than the other groups of compounds. Matt suggested looking at the SAR diagrams of these compounds to see if there are any common patterns.
However, Nuvisan also ran LIPR2 assays, which found that the unrelated lipase LIPR2 was inhibited by almost all of the compounds (Note: Eve has not yet discussed this result with Anna, who ran the assay). Madison commented that the fact that LIPR2 is inhibited by so many compounds is biochemically bizarre. She suggests that perhaps there is a contaminant or buffer component that is being carried over in every test that is causing this inhibition. It will be worth looking closer at this.
Madison is performing SPR on these compounds which together with the other assays will help identify any compounds worth further investigation. She has identified a good buffer and has found that the protein is sticking well to the chip and is behaving well. She has tested the Nuvisan compound and say that it has the characteristic “quick on, quick off” binding behavior, making it a good positive control. She has started going through the ASMS hits but the most she has seen so far is sticky binding (slow on, doesn’t come off). This could mean the compound is non-specific, or it could be a specific binder with poor binding.
Madison has also been running DSF experiments to see if there is temperature stabilization, but has not seen any evidence of this yet. She comments that it might not be best assay for our purposes because to her knowledge neither she nor Nuvisan have tested a compound that has shown stabilization in a thermal shift assay. Instead, if there are any strong SPR hits, she will set up crystal trays to try to get the structures.
HitGen have finished the DEL screen of chicken PLCz1 and the data is being analyzed at UNC. Eve and Madison have also started drafting a publication of the PLCz1 results.
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 29 May 2025, 2pm London time