JH-RE-06

Catalog No.S8850 Batch:S885002

Technical Data

Formula	C₂₀H₁₆Cl₃N₃O₄
Molecular Weight	468.72	CAS No.	1361227-90-8
Solubility (25°C)*	In vitro	DMSO	40 mg/mL (85.33 mM)
		Water	Insoluble
		Ethanol	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description

JH-RE-06 is a potent REV1-REV7 interface inhibitor with an IC50 of 0.78 μM and Kd value of 0.42 μM, disrupting REV1-POL ζ-mediated mutagenic translesion synthesis (TLS).

Targets

REV1-REV7 ^[1] (Cell-free assay)	REV1-REV7 ^[1] (Cell-free assay)
0.42 μM(Kd)	0.78 μM

In vitro

JH-RE-06 disrupts mutagenic TLS by preventing recruitment of mutagenic POL ζ. Remarkably, JH-RE-06 targets a nearly featureless surface of REV1 that interacts with the REV7 subunit of POL ζ. Binding of JH-RE-06 induces REV1 dimerization, which blocks the REV1-REV7 interaction and POL ζ recruitment. JH-RE-06 inhibits mutagenic TLS and enhances cisplatin-induced-toxicity in cultured human and mouse cell lines.^[1]

In vivo

JH-RE-06 improves tumor cell response to cisplatin in vivo. Co-administration of JH-RE-06 with cisplatin suppresses the growth of xenograft human melanomas in mice, establishing a framework for developing TLS inhibitors as a novel class of chemotherapy adjuvants.^[1]

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines HT1080 cells, A375 cells, KP cells, MEF, LNCap cells, AG01522 cells. Concentrations 1.5 μM Incubation Time 24 h Method Clonogenic survival assay—300 cells are plated in triplicate in 6-well plates for 24 hours. Cisplatin is added to relevant wells for 24 hours. All plates are incubated at 37℃ for 24 hours. Media are changed the next day and in fresh media JH-RE-06 (at 1.5 μM concentration) is added to untreated or cisplatin-treated cells for another 24 hours. Media are changed at the end of these combination treatments, and cells are allowed to recover for 7 days. To stain the resulting colonies, media are aspirated and the fixative (50% methanol and 10% glacial acetic acid) is added for 10 minutes, followed by the addition of 0.02% Coomassie brilliant blue R-250 stain in methanol: acetic acid: water in a ratio of 46.5:7:46.5 (v/v/v). Colonies that stained blue and contained at least 40 cells are counted. Relative cell survival or colony formation is calculated by dividing colony counts from treated samples by the DMSO or untreated controls.
Animal Study:^{^[1]}	Animal Models 6–8-week-old NCRNU-F (nude) female mice Dosages 1.6 mg/kg Administration intra tumor injection

Cell Assay:^{^[1]}

Cell lines
HT1080 cells, A375 cells, KP cells, MEF, LNCap cells, AG01522 cells.
Concentrations
1.5 μM
Incubation Time
24 h
Method
Clonogenic survival assay—300 cells are plated in triplicate in 6-well plates for 24 hours. Cisplatin is added to relevant wells for 24 hours. All plates are incubated at 37℃ for 24 hours. Media are changed the next day and in fresh media JH-RE-06 (at 1.5 μM concentration) is added to untreated or cisplatin-treated cells for another 24 hours. Media are changed at the end of these combination treatments, and cells are allowed to recover for 7 days. To stain the resulting colonies, media are aspirated and the fixative (50% methanol and 10% glacial acetic acid) is added for 10 minutes, followed by the addition of 0.02% Coomassie brilliant blue R-250 stain in methanol: acetic acid: water in a ratio of 46.5:7:46.5 (v/v/v). Colonies that stained blue and contained at least 40 cells are counted. Relative cell survival or colony formation is calculated by dividing colony counts from treated samples by the DMSO or untreated controls.

Animal Study:^{^[1]}

Animal Models
6–8-week-old NCRNU-F (nude) female mice
Dosages
1.6 mg/kg
Administration
intra tumor injection

References

[1] Jessica L Wojtaszek, et al. Cell. 2019 Jun 27;178(1):152-159.e11.

Selleck's JH-RE-06 has been cited by 2 publications

POLQ seals post-replicative ssDNA gaps to maintain genome stability in BRCA-deficient cancer cells [ Mol Cell, 2022, S1097-2765(22)01070-X]	PubMed: 36455556
Translesion DNA synthesis mediates acquired resistance to olaparib plus temozolomide in small cell lung cancer [ Sci Adv, 2022, 8(19):eabn1229]	PubMed: 35559669

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SHIPPING AND STORAGE
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