NCT-505 (NCT 505; NCT505) is a novel and potent aldehyde dehydrogenase 1A1 (ALDH1A1) inhibitor with potential anticancer activity. Overexpression of certain ALDHs (e.g., ALDH1A1) is an important biomarker in cancers and cancer stem cells (CSCs). NCT-505 inhibited the formation of 3D spheroid cultures of OV-90 cancer cells, and potentiated the cytotoxicity of paclitaxel in SKOV-3-TR, a paclitaxel resistant ovarian cancer cell line.

Physicochemical Properties

Molecular Formula	C27H28FN5O3S
Molecular Weight	521.606328010559
Exact Mass	521.189
CAS #	2231079-74-4
Related CAS #	2231079-74-4;2231079-75-5 (TFA);
PubChem CID	134821685
Appearance	White to off-white solid powder
LogP	2.6
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	4
Heavy Atom Count	37
Complexity	972
Defined Atom Stereocenter Count	0
SMILES	S(C)(N1CCN(C(C2=CN=C3C=CC(=CC3=C2N2CCC(C#N)(C3C=CC=CC=3)CC2)F)=O)CC1)(=O)=O
InChi Key	XMNLWOQHMQRHDV-UHFFFAOYSA-N
InChi Code	InChI=1S/C27H28FN5O3S/c1-37(35,36)33-15-13-32(14-16-33)26(34)23-18-30-24-8-7-21(28)17-22(24)25(23)31-11-9-27(19-29,10-12-31)20-5-3-2-4-6-20/h2-8,17-18H,9-16H2,1H3
Chemical Name	1-(6-Fluoro-3-(4-(methylsulfonyl)piperazine-1-carbonyl)-quinolin-4-yl)-4-phenylpiperidine-4-carbonitrile
Synonyms	NCT 505 NCT-505NCT505
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	IC50: 7 nM (ALDH1A1), >57 μM (hALDH1A2), 22.8 μM (hALDH1A3), 20.1 μM (hALDH2), >57 μM (hALDH3A1)
ln Vitro	Compound 86, also known as NCT-505, exhibits robust and selective activity against aldehyde dehydrogenase (ALDH1A1), with an IC50 of 7 nM. It also exhibits modest inhibition against hALDH1A2, hALDH1A3, hALDH2, and hALDH3A1 (IC50s, >57, 22.8, 20.1, >57 μM). NCT-505 does not appear to have any discernible inhibitory effects on type-4 hydroxysteroid dehydrogenase (HSD17β4) and 5-hydroxyprostaglandin dehydrogenase (HPGD) (IC50, >57 μM). Furthermore, NCT-505 exhibits strong cellular activities that lower OV-90 cell viability at an EC50 of 2.10-3.92 μM. Moreover, NCT-505 is cytotoxic to SKOV-3-TR cells, as demonstrated by IC50 values in the titration assay of 1, 3, 10, 20, and 30 μM, respectively[1].
Cell Assay	Cells are harvested, and an equal volume of first compound (NCT-505 or paclitaxel (Taxol)) at the indicated concentration or vehicle DMSO (final DMSO concentration is the same in all conditions) is added to the cell suspension before dispensing. Cells are dispensed into 384-well, white, TC-treated plates at a density of 3000 cells/well in a volume of 30 μL of growth media/well using a Multidrop Combi dispenser. Immediately after dispensing, the second compound (ALDH1A1 inhibitor or paclitaxel) and control solutions (92 nL) are transferred using a pintool. Plates are covered with a breathable seal and incubated for 4 days at 37°C, 5% CO2, 85% RH followed by addition of 20 μL of CellTiter-Glo. After a ∼30 min incubation at rt, samples are analyzed for luminescence intensity using a ViewLux high-throughput CCD imager equipped with clear filters. Pinned compounds are tested as 16-point dilution series, with concentrations ranging from 30.7 μM to 70.1 nM for ALDH1A1 inhibitors (NCT-505, etc.) or 31.7 μM to 0.034 nM for paclitaxel, in triplicate. Data are normalized to positive control bortezomib (1 μM final) and neutral control DMSO.
References	[1]. Discovery of Orally Bioavailable, Quinoline-Based Aldehyde Dehydrogenase 1A1 (ALDH1A1) Inhibitors with Potent Cellular Activity. J Med Chem. 2018 Jun 14;61(11):4883-4903.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~191.71 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.79 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 (4.79 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9171 mL	9.5857 mL	19.1714 mL
	5 mM	0.3834 mL	1.9171 mL	3.8343 mL
	10 mM	0.1917 mL	0.9586 mL	1.9171 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.