RSM-932A (TCD-717) is a specific ChoKα inhibitor (antagonist) with IC50 of 1 and 33 μM for human recombinant ChoKα and ChoKβ enzymes, respectively. RSM-932A is a “first-in-human” compound targeting ChoKα. RSM-932A has potent anti-tumor proliferation in vitro and anti-tumor activity in vivo, and has low toxicity [3].

Physicochemical Properties

Molecular Formula	C46H38BR2CL2N4
Molecular Weight	877.546
Exact Mass	874.084
Elemental Analysis	C, 62.96; H, 4.36; Br, 18.21; Cl, 8.08; N, 6.38
CAS #	850807-63-5
PubChem CID	11239948
Appearance	White solid powder
Melting Point	255 - 257 °C
LogP	13.09
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	9
Heavy Atom Count	54
Complexity	990
Defined Atom Stereocenter Count	0
InChi Key	DGBVSSXGHKAASQ-UHFFFAOYSA-L
InChi Code	InChI=1S/C46H38Cl2N4.2BrH/c1-49(39-23-19-37(47)20-24-39)43-27-29-51(45-9-5-3-7-41(43)45)31-33-11-15-35(16-12-33)36-17-13-34(14-18-36)32-52-30-28-44(42-8-4-6-10-46(42)52)50(2)40-25-21-38(48)22-26-40;;/h3-30H,31-32H2,1-2H3;2*1H/q+2;;/p-2
Chemical Name	1-[[4-[4-[[4-(4-chloro-N-methylanilino)quinolin-1-ium-1-yl]methyl]phenyl]phenyl]methyl]-N-(4-chlorophenyl)-N-methylquinolin-1-ium-4-amine;dibromide
Synonyms	RSM-932A; RSM932A; RSM-932A; 850807-63-5; RSM 932A; UNII-KC1UXA6R4Z; KC1UXA6R4Z; 1,1'-([1,1'-biphenyl]-4,4'-diylbis(methylene))bis(4-((4-chlorophenyl)(methyl)amino)quinolin-1-ium) bromide; Choline kinase alpha inhibitor TCD-717; TCD-717; RSM 932A
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 (e.g. under nitrogen), avoid exposure to moisture and light.
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	ChoKα(IC50 = 1 and 33 μM for ChoKα and ChoKβ)
ln Vitro	Cell line RSM-932A (TCD-717; 2-4 μM; 24 hours) promotes cell death in thermal insulation. RSM-932A has strong antiproliferative activity against most derived cell lines tested, including those from breast tumor gland, lung, bladder, bladder, cervix, bladder, melanoma, and brain tumors [2]. the 72-hour IC50 is 1.3-7.1 μM[1]. With an IC50 of 0.5 μM, RSM-932A (2–10 μM) inhibits Streptococcus pneumophila choline kinase (sChoK); quantifiable thoracic acid synthase (TS) and thoracic acid synthase (TK1) protein were demonstrated colorimetrically and by LDH/PK[2]. RSM-932A has a minimum inhibitory concentration (MIC) of 0.4 μM and a minimum lethal concentration (MLC) of 1.6 μM against Streptococcus pneumoniae [3]. In terms of enzyme determination, RSM-932A is a reasonably effective control [3]. The IC50 is 1.75 μM in a batch process where the concentration of choline is equal to its Km [4].
ln Vivo	RSM-932A is non-toxic at dosages that are efficacious and demonstrates strong in vivo anticancer activity [1].
Enzyme Assay	Enzymatic activity assays[1] Recombinant bacterial extracts, in which it has been introduced the expression of human ChoKα or ChoKβ, were used. The enzymatic reaction in 50 μL total volume contained a constant volume of bacterial extract in the presence of 100 mmol/L Tris-HCl, pH 8.0, 100 mmol/L MgCl2, and 10 mmol/L ATP, 200 μmol/L choline and methyl-[14C]-choline, and different concentrations of the drug. The reaction was incubated 30 minutes at 37°C and stopped on ice. The samples were solved by thin layer chromatography (TLC 60A Silica Gel Whatman). The radioactivity corresponding to phosphocholine was quantified with the Cyclone Plus Scanner. Concentration of each compound that produces 50% inhibition was calculated and expressed in μmol/L (IC50).
Cell Assay	Cell Proliferation Assay[2] Cell Types: DLD-1, HT29, SW620 and HCT116 CRC cell lines and non-tumorigenic CCD-841 cell line Tested Concentrations: 2, 3, 4 µM Incubation Duration: 24 hrs (hours) Experimental Results: Triggered cell death. Western Blot Analysis[2] Cell Types: DLD-1, HT29 and SW620 Cell lines Tested Concentrations: 2, 4, 6, 8, 10 uM Incubation Duration: 24 hrs (hours) Experimental Results: Thymidylate synthase (TS) levels are dose-dependent diminished) and thymidine kinase (TK1) protein.
Animal Protocol	Animal/Disease Models: Athymic nu/nu (nude) mice, CD1 nude mice and BALB/c nude mice (six weeks old) bearing human tumor xenografts (colon adenocarcinoma HT29, non-small cell lung cancer (NSCLC) H-460, breast cancer MDA-MB-231)[3] Doses: 7.5 mg/kg, 6 mg/kg, 5 mg/kg, 3 mg/kg, 1 mg/kg, 0.3 mg/kg Route of Route of Administration: Route of Administration (intraperitonealor intravenously (iv) (iv)(iv)), treatment regimen (5 days, 3 days per week, 2 days per week, 1 day per week) Experimental Results: LD50 was 10.9 mg/kg in mice. The effective dose used in in vivo experiments was 7.5 mg/kg.
References	[1]. Juan Carlos Lacal, et al. Preclinical characterization of RSM-932A, a novel anticancer drug targeting the human choline kinase alpha, an enzyme involved in increased lipid metabolism of cancer cells. Mol Cancer Ther. 2015 Jan;14(1):31-9. [2]. Ana de la Cueva, et al. Combined 5-FU and ChoKα inhibitors as a new alternative therapy of colorectal cancer: evidence in human tumor-derived cell lines and mouse xenografts. PLoS One. 2013 Jun 10;8(6):e64961. [3]. Tahl Zimmerman, et al. Identification and validation of novel and more effective choline kinase inhibitors against Streptococcus pneumonia. Sci Rep. 2020 Sep 22;10(1):15418. [4]. Tahl Zimmerman, et al. Antiplasmodial activity and mechanism of action of RSM-932A, a promising synergistic inhibitor of Plasmodium falciparum choline kinase. Antimicrob Agents Chemother. 2013 Dec;57(12):5878-88.
Additional Infomation	Choline Kinase Alpha Inhibitor TCD-717 is a small-molecule inhibitor of choline kinase alpha (CHKA), with potential antineoplastic activity. TCD-717 targets and binds to CHKA, an enzyme that plays a key role in the synthesis of phosphatidylcholine, the major phospholipid in eukaryotic cell membranes. Blockade of this enzyme induces cells to activate a different route for phospholipid production which causes a toxic effect and eventually leads to cell destruction. CHKA, overexpressed in human cancer cells while only minimally expressed in normal cells, appears to play a significant role in cellular proliferation, evasion of apoptosis, increased cell motility and metastasis.

Solubility Data

Solubility (In Vitro)

DMSO : ~16.67 mg/mL (~19.00 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 1.67 mg/mL (1.90 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 16.7 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.1395 mL	5.6977 mL	11.3954 mL
	5 mM	0.2279 mL	1.1395 mL	2.2791 mL
	10 mM	0.1140 mL	0.5698 mL	1.1395 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.