LLY-507

Protein target name:

SMYD2

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

N/A

Probe Availability

Tocris (LLY507)

MedChem Express (1793053-37-8)

MilliporeSigma (LLY507)

Cayman (1793053-37-8)

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Protein methyltransferase

HUGO ID:

HGNC:20982

Entrez gene ID:

56950

UNIPROT ID:

Q9NRG4

Target alias:

KMT3C, HSKM-B, ZMYND14

Probe Details

Chemical Probes Portal ID:

CPP46

Probe alias:

GTPL8239, ZINC231558920, CS-5126, HY-19313

PubChem CID:

91623361

ChEMBL ID:

CHEMBL3414623

SMILES string:

CC1=CN(C2=CC=CC=C12)CCN3CCN(CC3)C4=CC=CC=C4C5=CC(=CC(=C5)C#N)C(=O)NCCCN6CCCC6

InChi Key:

PNYRDVBFYVDJJI-UHFFFAOYSA-N

Control compound:

SGC705

Orthogonal probe:

BAY-598

PAINs evaluation:

PAINs free

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Solubility - concentration:

5 mg/mL

Storage Recommendations

Storage (solution):

2 years -20 oC

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

15 nM

Potency assay:

Scintillation proximity assay

PDB ID for probe-target interaction (3D structure):

4WUY

Off target activity

Potency assay (off target):

No significant activity was detected against 25 protein methyltransferases or DNMT in SPA, with LLY-507 100-fold selective for SMYD2>SMYD3, KMT5B and KMT5C.

Probe Selectivity in Vitro:

Profiled against 454 kinases in the DiscoveRx Kinome scan assay, as well as 36 GPCRs, 14 nuclear hormone receptors and 3 P450 enzymes using Eurofins-CEREP. LLY-507 inhibited 98% of ADRA1A activity at 10 uM.

In cell validation

On target activity in cells

Potency in cells:

IC50

0.6 uM

Potency assay (cells):

Protein substrate methylation assay in SMYD2-overexpressing U2OS cells with substrate methylation detected by ELISA.

Target engagement assay (cells):

Indirect, substrate methylation assay noted above plus several other related assays.

Primary Reference

Reference (doi):

10.1074/jbc.M114.626861

Reference (PMID):

25825497

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

no rating

SAB Members

SAB Comments

LLY-507 was disclosed as a potent and selective small molecule inhibitor of SMYD2 with cellular target engagement by inhibition of SYMD2 mediated monomethylation of p53 K370. LLY-507 inhibits the proliferation of ESCC, HCC and breast cancer cell lines. LLY-507 was initially shown to be selective (>100-fold) over a panel of representative methyltransferases, kinases, GPCRs and NHRs. However, recent additional screening at the NIMH Psychoactive Drug Screening program has shown LLY-507 to have potent inhibition of several targets (14 targets with Ki <1 uM and 3 targets with Ki <100 nM). These off-target activities of LLY-507 could confound results in cell assays by showing a response not related to inhibition of SMYD2 and potentially have unwanted psychoactive effects in animal models. Hence, it is recommended that an alternative SMYD2 chemical probe is used, such as A-893, AZ-505, or BAY-598; ideally use all three in parallel to ensure a consistent outcome to confidently link the response to inhibition of SMYD2 activity.

Jun 30 2016 - 4:01am

LLY-507 shows good potency and specificity for SMYD2 compared with other protein methyltransferases as well as GPCRs and protein kinases in assays.

Sep 6 2016 - 10:17am

LLY-507 potently and selectively inhibits the protein methyltransferase SMYD2 (IC50 <15 nM in an enzyme assay) compared with other pharmacologically important proteins including other HMTs, protein kinases, GPCRs and NHRs. At microM concentrations, the compound can be used in cellular assays (IC50=0.6 uM in U2OS cells for p53 methylation). However, it is preferable to simultaneously use other chemical probes or knock-down technologies for validations studies, because bona fide physiological substrates and functions of SMYD2 are still unknown and enigmatic.

Sep 7 2016 - 5:42am

LLY-507 is first-in-class inhibitor of SMYD2, where the biology and mechanism of action are poorly understood. It is quite potent (IC50 = 15 nM) in the the scintillation proximity assay; however, in the protein substrate assay, the IC50 = 600 nM. LLY-507 has a high degree of selectivity when screened against other targets like SMYD3 and SUVH420H (other protein or DNA methyltransferases). The inhibitor gives the same phenotype that was seen in the SMYD2 knockdown model. So far, the mechanism of action has been linked to the inhibition of SMYD2-mediated methylation of p53 K370 in cells. A co-crystal structure of LLY-507 and SMYD2 was obtained, and the structure was solved with a resolution near 1.63 angstroms. The structure will prove useful for designing the next generation of inhibitors. Very brief information was given about the stability of LLY-507, indicating stability after storage at -20 C for 2 years. No pharmacokinetic data were presented for animal exposure. The synthetic pathway to this molecule is thoroughly described in the reference with a path forward and few synthesis issues.

Sep 12 2016 - 6:49pm

LLY-507

Probe Availability

Probe Information

Target Details

KMT3C, HSKM-B, ZMYND14

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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LLY-507

Probe Availability

Probe Information

Target Details

KMT3C, HSKM-B, ZMYND14

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?