PFM01 is a nuclease-specific MRE11 inhibitor. PFM01 selectively inhibits MRE11 endo-but not exo-nuclease activity by targeting MRE11 at a location close to the dimer interface that is different from the ones that Mirin and PFM39 occupy. This allows for the disruption of the ssDNA-binding groove. Within the MRE11-RAD50-NBS1 (MRN) complex, MRE11 functions in signaling, detection, and DNA double-strand break repair (DSBR).

Physicochemical Properties

Molecular Formula	C14H15NO2S2
Molecular Weight	293.399
Exact Mass	293.054
Elemental Analysis	C, 57.31; H, 5.15; N, 4.77; O, 10.91; S, 21.85
CAS #	1558598-41-6
Related CAS #	1558598-41-6
Appearance	Yellow to orange solid powder
Density	1.4±0.1 g/cm3
Boiling Point	435.2±55.0 °C at 760 mmHg
Flash Point	217.0±31.5 °C
Vapour Pressure	0.0±1.1 mmHg at 25°C
Index of Refraction	1.684
LogP	2.74
SMILES	S1/C(=C\C2=CC=C(O)C=C2)/C(=O)N(CC(C)C)C1=S
InChi Key	GPURHDUTZUYAFI-GHXNOFRVSA-N
InChi Code	InChI=1S/C14H15NO2S2/c1-9(2)8-15-13(17)12(19-14(15)18)7-10-3-5-11(16)6-4-10/h3-7,9,16H,8H2,1-2H3/b12-7-
Chemical Name	(5Z)-5-[(4-hydroxyphenyl)methylidene]-3-(2-methylpropyl)-2-sulfanylidene-1,3-thiazolidin-4-one
Synonyms	PFM01; PFM-01; PFM 01
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	MRE11
ln Vitro	PFM01 significantly lessens the double-strand break (DSB) repair defect caused by mirin or PFM39 in irradiated G2 cells.It also rescues the repair defect in 48BR (WT) and HSC62 (BRCA2-defective) primary cells, diminishes the formation of RAD51 foci in 1BR3 (WT) and HSC62 (BRCA2-defective) cells, enhances non-homologous end-joining (NHEJ) in H1299 dA3 cells, and decreases homologous recombination (HR) in U2OS DR-GFP cells.[1]
Cell Assay	The following fibroblast types were cultured in Dulbecco's modified Eagles medium: 48BR (human primary fibroblasts), ATLD2 (primary fibroblasts), 1BR3 hTERT, HSC62 hTERT (BRCA2-defective), and 2BN hTERT (XLF-defective) (12%). In minimal essential medium (MEM) supplemented with 10% FCS, A549 cells were cultivated. The medium was exposed to 137Cs radiation on the cells. The concentrations of MRE11 inhibitors were, unless specified otherwise, 500 μM Mirin, 100 μM PFM39, 100 μM PFM01, and 100 μM PFM03. 30 minutes before irradiation, either 10 μM ATM inhibitor (KU55933) or 10 μM DNA-PKcs inhibitor (NU7441) were added, unless otherwise specified.
References	[1]. DNA double-strand break repair pathway choice is directed by distinct MRE11 nuclease activities. Mol Cell. 2014 Jan 9; 53(1): 7-18. [2]. RAD50 regulates mitotic progression independent of DNA repair functions. FASEB J. 2020 Feb; 34(2): 2812-2820.

Solubility Data

Solubility (In Vitro)

DMSO: ≥ 100 mg/mL (~340.8 mM) Ethanol: ~25 mg/mL (~85.2 mM)

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.4083 mL	17.0416 mL	34.0832 mL
	5 mM	0.6817 mL	3.4083 mL	6.8166 mL
	10 mM	0.3408 mL	1.7042 mL	3.4083 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.