RHPS4 methosulfate (also known as NSC714187) is a potent inhibitor of telomerase with submicromolar activity. RHPS4 treatment of UXF1138L cells resulted in chromosome fusions, telomere-initiated DNA damage signaling, and the removal of the telomerase catalytic subunit (hTERT) from the nucleus. It was also reported that RHPS4 exhibited greater potency against cancer cells growing as colonies as opposed to monolayers.

Physicochemical Properties

Molecular Formula	C23H20F2N2O4S
Molecular Weight	458.48
Exact Mass	458.11
Elemental Analysis	C, 60.25; H, 4.40; F, 8.29; N, 6.11; O, 13.96; S, 6.99
CAS #	390362-78-4
Related CAS #	390362-77-3 (cation); 390362-78-4 (methosulfate)
PubChem CID	9804187
Appearance	Brown to reddish brown solid powder
Melting Point	256-258 ºC
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	0
Heavy Atom Count	32
Complexity	633
Defined Atom Stereocenter Count	0
SMILES	C1C(F)=CC=C2N(C)C3=CC(C)=CC4=C5C=C(F)C=CC5=[N+](C)C(=C34)C=12.S(=O)([O-])(=O)OC
InChi Key	VRWGYMXWYZBBGF-UHFFFAOYSA-M
InChi Code	InChI=1S/C22H17F2N2.CH4O4S/c1-12-8-16-15-10-13(23)4-6-18(15)26(3)22-17-11-14(24)5-7-19(17)25(2)20(9-12)21(16)22;1-5-6(2,3)4/h4-11H,1-3H3;1H3,(H,2,3,4)/q+1;/p-1
Chemical Name	4,16-difluoro-8,11,20-trimethyl-8-aza-20-azoniapentacyclo[11.7.1.02,7.09,21.014,19]henicosa-1(20),2(7),3,5,9,11,13(21),14(19),15,17-decaene;methyl sulfate
Synonyms	NSC714187; NSC 714187; NSC-714187; RHPS 4 methosulfate; RHPS-4; RHPS 4; RHPS4
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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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	telomerase in the TRAP assay ( IC50 = 0.33 μM )
ln Vitro	In vitro activity: RHPS4 causes a fast and potent DNA damage response at telomeres in melanoma and human transformed fibroblast cells, which is typified by the development of multiple telomeric foci with phosphorylated RAD17, 53BP1, and γ-H2AX DNA damage response factors. DNA repair enzyme ATR is required for this, and overexpression of POT1 or TRF2 will counteract it. It is also linked to the delocalization of the protective telomeric DNA-binding protein POT1.
ln Vivo	RHPS4 caused telomere damage and tumor cell apoptosis in mice to produce its antitumor effect on xenografts of human tumor cells with varying histopathologies. Strong DNA damage response was observed in conjunction with tumor inhibition, and RHPS4 treatment was ineffective against tumors overexpressing POT1 or TRF2[2].
Cell Assay	Twenty-four hours after plating, RHPS4 is added to the cells at different times and at a concentration of 1 μM. Every experiment counts the number of cells and determines their viability (trypan blue dye exclusion). For 24 hours, caffeine at a dose of 10 mM—which has no harmful effects on cell viability—is added to the medium. For thirty minutes, bleomycin is used at a concentration of 30 mM.
Animal Protocol	Dissolved in saline; 15 mg/kg/d; i.v. injection CD-1 male nude (nu/nu) mice
References	[1]. Br J Cancer . 2007 Apr 23;96(8):1223-33. [2]. J Clin Invest . 2007 Nov;117(11):3236-47.

Solubility Data

Solubility (In Vitro)

DMSO: 1~91 mg/mL (2.2~198.5 mM) Water: <1 mg/mL Ethanol: <1 mg/mL

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	2.1811 mL	10.9056 mL	21.8112 mL
	5 mM	0.4362 mL	2.1811 mL	4.3622 mL
	10 mM	0.2181 mL	1.0906 mL	2.1811 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.