KRAS switch I/II pocket inhibitor | BI-2852
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Highlights
BI-2852 is a potent inhibitor for in vitro use that directly targets GTP-bound KRAS, which is the major form present in cancer cells carrying KRAS mutations. BI-2852 binds to KRASG12D with a KD of 740 nM (ITC), inhibits GTP-KRASG12D binding to effectors like SOS1, CRAF and PI3Kα with an IC50 of 490, 770 and 500 nM. BI-2852 showed pERK(2h) modulation and antiproliferative effects in KRASmut cells (NCI-H358) at 5.8 µM and 6.7 µM.5
Together with the structurally similar negative control BI-2853 which is also available via opnMe.com, the probe BI-2852 can serve as an excellent small molecule tool inhibitor of KRAS for testing biological hypotheses in vitro.
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Target information
The three human RAS genes, KRAS, NRAS and HRAS encode four different RAS proteins (KRAS-4A, KRAS-4B, NRAS and HRAS) which belong to the protein family of small GTPases. The RAS proteins function as molecular switches between active GTP-bound and inactive GDP-bound conformations. RAS is the most frequently mutated oncogene in human cancers (~27%) with activating mutations mainly in codons 12, 13 and 61. The main mutation in codon 12 causes RAS activation by interfering with GAP binding and GAP-stimulated GTP hydrolysis. KRAS mutations rates are high in pancreatic (~90%), colorectal (~45%) and lung adenocarcinomas (~35%).1,2 KRAS could serve as an excellent drug target for many cancers, but direct inhibition of oncogenic RAS has proven to be challenging. After more than three decades of intense effort, the first anti-RAS therapies have just reached clinical application in the beginning of 2019.1-5
Complex of KRAS with BI-2852
In vitro Activity
BI-2852 displays an IC50 of 490 nM in a GTP-KRASG12D::SOS1 AlphaScreen (AS) assay leading to low micromolar inhibition of pERK (in H358 cell line)5.
|
Probe name / Negative control |
BI-2852 |
BI-2853 |
|
MW [Da] |
516.6 |
516.6 |
| ITC (KD) GCP-KRASG12D [µM]a |
0.74 |
n.d. |
| ITC (KD) GCP-KRASwt [µM]a |
7.5 |
n.d. |
| ITC (KD) GDP-KRASG12D [µM]a |
2.0 |
n.d. |
| ITC (KD) GDP-KRASwt [µM]a |
1.1 |
n.d. |
|
AS (IC50) GTP-KRASG12D::SOS1 [nM]a |
490 |
4400 |
|
AS (IC50) GTP-KRASG12D::CRAF [nM]a |
770 |
n.d. |
|
AS (IC50) GTP-KRASG12D::PI3Kα [nM]a |
500 |
n.d. |
|
AS (IC50) GDP-KRASG12D::SOS1 [nM]a |
260 |
2500 |
|
AS (IC50) GTP-KRASwt::SOS1 [nM]a |
490 |
n.d. |
|
EC50 pERK H358 cells (2 h) [µM]a |
5.8 |
> 50 |
|
EC50 H358 cells (low serum) [µM]a |
6.7 |
n.d. |
a for assay conditions please refer to reference 5
In vitro DMPK and CMC parameters
|
Probe name / Negative control |
BI-2852 |
BI-2853 |
|
logP |
2.6 |
2.6 |
|
Solubility @ pH 6.8 [µg/ml] |
18 |
21 |
|
CACO permeability @ pH 7.4 [*10-6 cm/s] |
5.0 |
<1.15 |
|
CACO efflux ratio |
2.1 |
n.d. |
|
MDCK permeability Pappa-b/b-a @ 1µM [10-6 cm/s] |
n.d. |
n.d. |
|
MDCK efflux ratio |
n.d. |
n.d. |
|
Microsomal stability (human/mouse/rat) [% QH] |
91 / 93 / 90 |
92 / 95 / 86 |
|
Hepatocyte stability (human/mouse/rat) [% QH] |
12 / 21 / 25 |
12 / 69 / 52 |
|
Plasma protein binding (human/mouse/rat) [%] |
98.8 / 99.5 / 98.5 |
98.7 / 99.1 / 98.6 |
|
CYP 3A4 (IC50) [µM] |
4.4 |
n.d. |
|
CYP 2C8 (IC50) [µM] |
8.4 |
n.d. |
|
CYP 2C9 (IC50) [µM] |
4.8 |
n.d. |
|
CYP 2C19 (IC50) [µM] |
11.0 |
n.d. |
|
CYP 2D6 (IC50) [µM] |
15.0 |
n.d. |
In vivo DMPK parameters
No data available, BI-2852 is an in vitro tool compound.
In vivo pharmacology
No data available, BI-2852 is an in vitro tool compound
Negative control
BI-2853 is the less active enantiomer of BI-2853. It shows no effect on cells and is around 10-fold less potent in the AS assays.
BI-2853 which serves as a negative control
Selectivity
Selectivity data are not available at this point in time.
|
Bi-3663 |
SELECTIVITY DATA AVAILABLE |
|
Cerep® |
No |
|
Panlabs® |
No |
|
Invitrogen® |
No |
|
DiscoverX® |
No |
|
Dundee |
No |
Co-crystal structure of the BI probe compound and the target protein
The Xray crystal structure of target in complex with BI-2852 is available (PDB code: 6GJ7)5.
Summary
The in vitro tool compound BI-2852 is a potent nanomolar inhibitor of the KRAS switch I/II pocket and directly inhibits both the active and inactive forms of KRAS.
Supplementary data
References
Targeting Mutant KRAS for Anticancer Therapeutics: A Review of Novel Small Molecule Modulators
Wang Y., Kaiser C. E., Frett B., and Li H.
J. Med. Chem. 2013, 56, 5219−5230.
Drugging the undruggable RAS: Mission possible?
Cox D., Fesik S. W., Kimmelman A. C., Luo J., Der C. J.
Nat. Rev. Drug. Discov. 2014, 13, 828-851.
Dragging ras back in the ring
Stephen A. G., Esposito D., Bagni R. K., McCormick F.
Cancer. Cell Mar 17, 25(3), 272-281.
RAS isoforms and mutations in cancer at a glance
Hobbs G. A., Der C. J., Rossman K. L.
J. Cell Science 2016, 129, 1287-1292.
Drugging an “undruggable” pocket on KRAS
Kessler D., Gmachl M., Mantoulidis A., Martin L. J., Zoephel A., Mayer M., Gollner A., Covini D., Fischer S., Gerstberger T., Gmaschitz T., Goodwin C., Greb P., Häring D., Hela W., Hoffmann J., Karolyi-Oezguer J., Knesl P., Kornigg S., Koegl M., Kousek R., Lamarre L., Moser F., Munico-Martinez S., Peinsipp C., Phan J., Rinnenthal J., Sai J., Salamon C., Scherbantin Y., Schipany K., Schnitzer R., Schrenk A., Sharps B., Siszler G., Sun Q., Waterson A., Wolkerstorfer B., Zeeb M., Pearson M., Fesik S. W. and McConnell D. B.
PNAS 2019, Latest articles, 1-7.
