CMLD-2

Protein target name:

ELAVL1

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

20 uM

Probe Availability

Sigma

Probe Information

Target and Probe Information

Target Details

Target class:

Other

Define other target class:

ELAVL family

Subclass:

RNA Binding Protein

HUGO ID:

HGNC:3312

Entrez gene ID:

1994

UNIPROT ID:

Q15717

Target alias:

HUR; Hua; MelG; ELAV1

Probe Details

Chemical Probes Portal ID:

CPP1870

PubChem CID:

16746438

ChEMBL ID:

CHEMBL1499653

SMILES string:

COC1C=C(OC)C(C(C2C=CC(OC)=CC=2)CC(=O)N2CCCC2)=C2C=1C(C1C=CC=CC=1)=CC(=O)O2

InChi Key:

PROGRNRRJJYCNX-UHFFFAOYSA-N

Control compound:

NC-3

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

0.35 uM

Potency assay:

FP assay, AlphaLISA assay, SPR

In cell validation

On target activity in cells

Potency assay (cells):

RNP IP, luciferase-based reporter assays, and cytotoxicity assays.

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

MTT-based cytotoxicity assay using human cancer cells, normal fibroblast cell line WI-38, and normal human colon epithelial cell line CCD 841 CoN. Moderate cytotoxicity against HCT-116 colon cancer cells and MiaPaCa2 pancreatic cancer cells. Decreased cytotoxicity against normal cell WI-38 and CCD 841 CoN as demonstrated by ∼2-fold greater IC50 values .

Primary Reference

Reference (doi):

10.1021/cb500851u 10.1038/s41598-017-07787-4 10.1038/s41598-019-43894-0

Reference (PMID):

25750985 28855578 31089242

Under Review

This probe is in the process of SAB review. Please continue to check back for new reviews and commentary.

SAB Rating

Rating for use in Cells

2 review(s)

Rating for use in Model Organisms

no rating

SAB Members

SAB Comments

CMLD-2 is a compound that emerged from a limited size HTS conducted to identify disruptors of binding between HuR protein (also known as ELAVL1) and mRNA adenine- and uridine-rich elements (ARE). The screening assay appears to have been appropriately validated and optimized as a part of the original publication (Wu et al., 2015).

MAJOR ISSUE: lack of selectivity data. CMLD-2 (PubChem CID: 16746438) was identified as a hit in other screening campaigns for other targets (for example as a Mcl-1/Noxa Interaction Inhibitor, and in other contexts). I don’t take this to mean conclusively that this compound is promiscuous but additional tests are needed to establish the full scope of targets. Additionally, it is not clear that the observed phenotype is HuR dependent, and more work needs to be done to establish that the observed phenotype is driven by CMLD-2 mediated disruption of HuR/ARE interaction. The lack of selectivity information precludes endorsement of CMLD-2 as a chemical probe (tool compound) for study of HuR (ELAVL1).

MINOR POINTS FOR CMLD-1 DEVELOPERS TO CONSIDER

The reported binding between HuR and ARE is on the order of low nM. The measured Ki of the lead screening hit CMLD-2 was 350nM. The hit compound was used without further optimization. I would advise a structure-guided med chem campaign to improve the in vitro binding while testing for in cell activity and target engagement at this point.
No SAR analysis. The HTS yielded a class of related compounds as hits but it is not clear whether any SAR trends can be rationalized.
The proposed negative control compound NC-3 retained some binding (2 uM) under same conditions. In cell viability assays in HCT-116, CMLD-2 IC50 was around 30 uM vs. 70 uM for NC-3. Similar was seen in MiaPaCa2 cells. Although other data did suggest that NC-3 is a much weaker inhibitor of HuR/ARE interaction, it is not a inactive, and therefore this calls into question whether NC-3 can be used as a negative control. I would advise caution here, and additional work on developing a better negative control compound. One might emerge from SAR work suggested above.

Jul 26 2020 - 10:02am

The molecule is chiral and lacks an enantiomeric comparison. Care using this probe must be taken as chromone template is known to interfere with spectrometric assays. The compound is reported as active in other RNA and DNA dependant assays in Pubchem Database.

Aug 28 2020 - 7:42am

CMLD-2

Probe Availability

Probe Information

Target Details

HUR; Hua; MelG; ELAV1

Probe Details

In vitro validation

In cell validation

Toxicity

Primary Reference

Supporting Organizations

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CMLD-2

Probe Availability

Probe Information

Target Details

HUR; Hua; MelG; ELAV1

Probe Details

In vitro validation

In cell validation

Toxicity

Primary Reference

Supporting Organizations

Interested in supporting the portal?