SCR7 (SCR-7; SCR 7) is a potent and selective inhibitor of DNA Ligase IV with potential anticancer activity. The enzyme DNA Ligase IV is in charge of using the non-homologous end joining (NHEJ) repair pathway to fix DNA double-strand breaks (DSB).

Physicochemical Properties

Molecular Formula	C18H12N4OS
Molecular Weight	332.38
Exact Mass	334.088
Elemental Analysis	C, 64.65; H, 4.22; N, 16.75; O, 4.78; S, 9.59
CAS #	1533426-72-0
Related CAS #	14892-97-8
PubChem CID	91885409
Appearance	Yellow solid powder
Density	1.3±0.1 g/cm3
Boiling Point	644.9±65.0 °C at 760 mmHg
Flash Point	343.8±34.3 °C
Vapour Pressure	0.0±2.0 mmHg at 25°C
Index of Refraction	1.675
LogP	-0.22
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	4
Heavy Atom Count	24
Complexity	570
Defined Atom Stereocenter Count	0
InChi Key	NEEVCWPRIZJJRJ-LWRDCAMISA-N
InChi Code	InChI=1S/C18H14N4OS/c23-17-15(19-11-13-7-3-1-4-8-13)16(21-18(24)22-17)20-12-14-9-5-2-6-10-14/h1-12H,(H2,21,22,23,24)/b19-11?,20-12+
Chemical Name	5-(benzylideneamino)-6-[(E)-benzylideneamino]-2-sulfanylidene-1H-pyrimidin-4-one
Synonyms	SCR-7; SCR 7; SCR7
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	DNA Ligase IV; CRISPR/Cas9
ln Vitro	SCR7 efficiently prevents the creation of multimers at concentrations greater than 200 μM. With IC50 values of 40, 34, 44, 8.5, 120, 10, and 50 μM, respectively, SCR7 effectively suppresses the growth of MCF7, A549, HeLa, T47D, A2780, HT1080, and Nalm6 cells. [1] SCR7 inhibits the NHEJ repair of DSBs produced by CRISPR-Cas9. [2]SCR7 enhances the effectiveness of CRISPR/Cas9-mediated HDR-mediated genome editing in mouse embryos and mammalian cells by up to 19 times[3].
ln Vivo	SCR7 treatment (10 mg/kg, i.m.) significantly decreases the tumor caused by breast adenocarcinoma and results in a 4-fold increase in longevity when compared to the control group. Nevertheless, neither tumor regression nor increased lifespan is seen in Swiss albino mice bearing the Dalton's lymphoma tumor model when SCR7 (20 mg/kg, i.p.) is administered. The cytotoxic effects of radiation, etoposide, and 3-Aminobenzamide on tumors derived from Dalton's lymphoma (DLA) cells are significantly enhanced in BALB/c mice when SCR7 (20 mg/kg, i.p.) is administered.[1]
Enzyme Assay	The oligomeric DNA substrates (5' compatible and 5' noncompatible ends) and increasing concentrations of purified Ligase IV/XRCC4 complex (30, 60, and 120 fmol) are added to the SCR7-treated extracts to conduct the complementation experiment. At 25°C, reactions are incubated for two hours. Following that, the reaction products are separated on an 8% denaturing PAGE. A PhosphorImager is used to detect the signal after the gel has dried and been exposed, and Multi Gauge (V3.0) software is used for analysis.
Cell Assay	MTT and trypan blue assays are used to measure the proliferation of cancer cells. In summary, SCR7 (10, 50, 100, and 250 μM) is added to MCF7, CEM, HeLa, A549, HT1080, A2780, T47D, Nalm6, N114, and K562 cells during 24 or 48 hours of growth before the cells are tested using MTT or trypan blue. Every experiment is conducted at least three separate times.
Animal Protocol	Animal/Disease Models: BALB/c mice bearing breast adenocarcinoma tumors [1] Doses: 10 mg/kg Route of Administration: Intraperitoneal injection/ip; twice a day Experimental Results: Significantly suppressed breast adenocarcinoma tumors and increased lifespan of animals. CRISPR components mixture (Cas9 mRNA, sgRNA and targeting template) and 10 mM of Scr7 NHEJ inhibitor (to 1 mM final) were injected into the cytoplasm at the pronuclear stage. Kell-LPETG mice
References	[1]. Cell . 2012 Dec 21;151(7):1474-87. [2]. Nat Biotechnol . 2015 May;33(5):543-8. [3]. Scientific Reports. 2017, 7, Article number: 8943.

Solubility Data

Solubility (In Vitro)

DMSO: ~66 mg/mL (~198.6 mM) Water: <1 mg/mL Ethanol: ~3 mg/mL (~9.0 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.48 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.48 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + + 45% Saline For example, if 1 mL of working solution is to be prepared, you can take 100 μL of 25 mg/mL of DMSO stock solution and add tO + 400 μL of PEG300, mix well (clear solution); Then add 50 μL of Tween 80 to the above solution, mix well (clear solution); Finally, add 450 μL of saline to the above solution, mix well (clear solution). Preparation of saline: Dissolve 0.9 g of sodium chloride in ddH ₂ O and make up to 100 mL to obtain a clear and transparent saline solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.0086 mL	15.0430 mL	30.0860 mL
	5 mM	0.6017 mL	3.0086 mL	6.0172 mL
	10 mM	0.3009 mL	1.5043 mL	3.0086 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.