VER-155008

VER-155008 is an Hsp70 inhibitor (IC50 = 0.5 μM), exhibiting over 100-fold selectivity against Hsp90, and capable of inhibiting Hsc70 and Grp78 (IC50 = 2.6 μM for both). This compound possesses good selectivity and cell permeability, and is used in anti-tumor research, showing therapeutic potential in leukemia and solid tumor models.

HSC70:2.6 μM, GRP78:2.6 μM, HSP70:0.5 μM

Methods: The in vitro activity of VER-155008 was validated in H1975 and H1299 cells using the MTS assay. Cells were incubated at a concentration of 10 μM for 24, 48, and 72 hours to detect its inhibitory effect on cell proliferation.
Results: VER-155008 effectively inhibited LUAD cell proliferation, and H1975 cells with high SPOCK1 expression showed higher sensitivity to it.[1]
Methods: Human anaplastic thyroid carcinoma cell lines 8505C and FRO were treated with 10–50 μM VER-155008 for 72 h, or with 50 μM for 24–72 h. Cell viability and the proportion of dead cells were detected using the CCK-8 assay and trypan blue staining.
Results: VER-155008 reduced cell viability and increased the proportion of dead cells in a time- and dose-dependent manner.[2]
Methods: Human NSCLC cell lines A549 and H1975 were incubated with 25 μM HSP70 inhibitor VER-155008 for 2 days. DNA synthesis was detected by BrdU incorporation assay, and cell viability was detected by MTT assay.
Results: VER-155008 treatment significantly reduced the percentage of BrdU-positive cells and cell viability, indicating its effective inhibition of NSCLC cell proliferation.[3]

Methods: The in vivo activity of VER-155008 was validated using a subcutaneous xenograft tumor model of LUAD cell lines. C57BL/6 mouse models were used, with intraperitoneal administration starting on day 7 after tumor cell injection, at a dose of 10 mg/kg every 2 days, with PBS as the vehicle.
Results: Tumor growth was significantly inhibited in the VER-155008 treatment group, and tumor weight was lower than that in the control group.[1]

Hsc70, Hsp70 and Grp78 ?uorescence polarisation (FP) assay: The FP assay for Hsp70 is conducted in aqueous buffer consisting of 100 mM Tris pH 7.4, 150 mM KCl and 5 mM CaCl2, in a final assay volume of 100 μl, using 96 well black polystyrene high bind plates with a Fusion plate reader. N6-(6-amino)hexyl-ATP-5-FAM and the in-house protein preparation of GST-HSP70 3-382 have final concentrations in the assay of 20 nM and 400 nM, respectively. Compounds are tested as 10-point IC50s, with a final DMSO concentration of 5%. Assay mixtures are incubated for 3 h prior to reading on the Fusion (ex 485 nm; em 535 nm). The data is fitted using a 4 parameter logistical data model by XLFit 4. The FP assay for Hsc70 and Grp78 is carried out as described for Hsp70 using the same N6 -(6-amino)hexyl-ATP-5-FAM as the FP probe with the following modi?cations. For Hsc70, the protein and probe concentrations are 0.3 μM and 20 nM, respectively with a 30 min incubation at 22℃ while for Grp78, the protein and probe concentrations are 2 μM and 10 nM, respectively with a 2 h incubation at 22℃. The KD for the FAM-ATP probe was 0.24 μM for Hsc70 and 2 μM for Grp78.

All cell lines are grown in DMEM/10% FCS with GlutaMAX-I in a humidified atmosphere of 5% CO2 in air. Cell proliferation is determined using the sulforhodamine B (SRB) assay.(Only for Reference)

H2O: < 1 mg/mL (insoluble)

DMSO: 79.17 mg/mL (142.29 mM), Sonication is recommended.

10% DMSO+40% PEG300+5% Tween 80+45% Saline: 2 mg/mL (3.59 mM), Sonication is recommended.