Screen3 - MMV active compounds 5-25

Optimize conditions for screen

screen works without problems at 10ul volume - mantis dispensing / 384well plate / 4 tips with 1ml 5+5 dispensed with mantis

Admin-Infos on MMV actives box 1

sent by Evotec (Tolouse/France). Organized by Elodie Chenu (MMV) chenue@mmv.org

Scientists at MMV

James Duffy duffyj@mmv.org / Jeremy Burrows burrowsj@mmv.org

Infos on MMV active box 1

Jeremy Burrows (burrowsj@mmv.org) email 16.4.25: MMV Actives Box 1 (AB1). This compound set comprises 352 hits from phenotypic NF54 screening of MMV libraries (actives are defined as IC50 ≤ 10 µM NF54, IC50 ≥ 10 µM HepG2). We do not know the targets of all of these compounds and, since the chemical structures are all attractive from a medicinal chemistry perspective, were you to screen and find a hit on Snf2L then that could help your project move forward.

5ul 10mM in DMSO; plates arrived 250508 box is in -80°C Lagerraum

Plate map - MMV - Actives Box2500215175.xls uploaded

first 22 columns are full (in the list 2 compounds are out of order - are at the end of the list)

How to progress with the 384 plate

Todo: inhibitors are at 10mM 5ul

• add our inhibitors to the last columns of the plate as well 10 mM final - add 5ul
  each 2wells for our 7 inhibitors (positive controls)
• add 6 DMSO controls 5ul
• make copies of the plate for the study:
  first round: 50uM Inhibitor; final volume of the reaction 12ul; 0.06ul of the inhibitor (0.5%DMSO)
  4 copy plates (2plates for pvSnf2L)
  second round: 20uM inhibitor; either for lower reaction volume or re-test; 0.025ul of the inhibitors (0.2%DMSO)
  2 copy plates (1plate for pvSnf2L)
• How many copies: for testing and playing - 4 copies +2 for a Pvivax test = 6 copies

Control Inhibitors for the 384well MMV plate (with their short name in blue)

from Screen2

1) Prestw-1472 (EC50 5-10uM) Hexachlorophene (XXX): I H1

from Screen2

2) NH125 (Sigma): I N1

3) S2303 (3uM - enantiomer S2812) Gossypol Acetate (Selleckchem): I G1

4) S2812 (R)-(-)-Gossypol (AT-101) acetic acid (Selleckchem): I G2

5) S2872 (5uM), GW5074 (Selleckchem): I G3

6) S7531 (2.5uM) / UMI-77 (Selleckchem): I U1

7) S2671 (15uM), AS-252424 (Selleckchem): I A1

Location of the control inhibitors on the MMV plate

DMSO A23, A24,

IH1 B23, B24

IN1 C23, C24

IG1 D23, D24

IG2 E23, E24

IG3 F23, F24

DMSO G23, G24

IU1 H23, H24

IA1 I23, I24

DMSO J23, J24

Plates for screening were plated by Clemens Entzian 6.6.25

Die Platten sind fertig und können abgeholt werden. Well A1 konnte bei destination plate 25061301, 25061302 und 25061303 nicht gespottet werden, da wahrscheinlich das DMSO zu viel Wasser gezogen hat. Diese 3 plates waren die für die 50µM Konzentration.

Pipetting scheme

Use the Mantis - Mantis Dispensor for Screen

372 reactions on the plate: K23-P23 and K24-P24 are free

372x12= 4464 total reaction volume

Reaction volume 12 ul split into: 372x3 = 1116ul PfSnf2L+ParM in buffer

372x9= 3348 DNA, ATP, Buffer

Reaction volume 10 ul: 384x10=3840ul

4 HV chips: je ca. 1 ml - 5ul DNA/ATP/ParM/Buffer +5ul Snf2L/Buffer

250715

250715: dispense program 10ul - 4 tips #1,2 HV to #4,5HV links 1,2 rechts 4,5 / 955ul in 1000er reicht aus alles zu fahren.

HV chips setting: no prime; 5ul pre-dispense

dispense file

MMV active plate screen Juli/August 2025

pfSnf2L fl fraction:xxxx

2x 50uM compound; 1x 20uM compound

pvSnf2Lcore fraction:xxxx

2x 50uM; 1x 20uM compound

Hits: L9, B9 PfSnf2L; H16, L9 PvSnf2L

Selected compounds for dose response plate

Maresa Inhibitors: all 7

MMV Active plate: XXX (12)

Concentration: 20uM, 7uM, 2uM (5 copies)