SP2509 (HCI2509) is a novel, potent and selective histone demethylase LSD1 (KDM1A) antagonist with antineoplastic activity. It inhibits LSD1 with an IC50 of 13nM, and shows no/little activity against MAO-A, MAO-B, lactate dehydrogenase and glucose oxidase. SP-2509 exhibits excellent antiproliferative activity and high in vivo antitumor efficacy. It showed high actitivity on human AML cells. Treatment with the novel LSD1 antagonist SP2509 attenuated the binding of LSD1 with the corepressor CoREST, increased the permissive H3K4Me3 mark on the target gene promoters, and increased the levels of p21, p27 and CCAAT/enhancer binding protein α in cultured AML cells.

Physicochemical Properties

Molecular Formula	C19H20CLN3O5S
Molecular Weight	437.90
Exact Mass	437.081
CAS #	1423715-09-6
Related CAS #	1423715-09-6
PubChem CID	135743577
Appearance	White to off-white solid powder
Density	1.4±0.1 g/cm3
Index of Refraction	1.645
LogP	3.64
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	5
Heavy Atom Count	29
Complexity	705
Defined Atom Stereocenter Count	0
SMILES	C/C(=N\NC(=O)C1=CC(=CC=C1)S(=O)(=O)N2CCOCC2)/C3=C(C=CC(=C3)Cl)O
InChi Key	NKUDGJUBIVEDTF-FYJGNVAPSA-N
InChi Code	InChI=1S/C19H20ClN3O5S/c1-13(17-12-15(20)5-6-18(17)24)21-22-19(25)14-3-2-4-16(11-14)29(26,27)23-7-9-28-10-8-23/h2-6,11-12,24H,7-10H2,1H3,(H,22,25)/b21-13+
Chemical Name	(E)-N'-(1-(5-chloro-2-hydroxyphenyl)ethylidene)-3-(morpholinosulfonyl)benzohydrazide.
Synonyms	SP 2509; SP-2509; SP2509.
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: This product requires protection from light (avoid light exposure) during transportation and storage.
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

ln Vitro	In cultured human acute myeloid leukemia cells, SP2509 (250, 500, and 1000 nM) suppresses LSD1 activity, reduces colony formation, and causes apoptosis and cell death. It also raises H3K4Me3 on the promoters of p57 Kip, KLF4, and p21 and stimulates mRNA expression of p57Kip, KLF4, and p21 in AML cells. Primary and grown AML cells exhibit characteristics of morphologic differentiation when exposed to SP2509 (250, 1000 nM). Additionally, SP2509 and PS work together to produce synergistic lethal action against both primary and cultured AML cells[1]. Both the CoREST-LSD1 interaction and the CRC are not significantly destabilized by SP2509. At low concentrations of 0.1 µM, SP2509 causes cell death, but no morphological alterations are seen. Similarly, SP2509 disrupts the ability of medulloblastoma cells to proliferate[2].
ln Vivo	The administration of SP2509 (25 mg/kg) and/or PS (5 mg/kg) to mice harboring AML xenografts and primagrafts significantly increases PS-mediated loss of viability of CD34+ primary AML cells[2].
Animal Protocol	Formulated in 20% Cremaphor, 20% DMSO, 60% sterile water; 25 mg/kg; i.p. injection NOD/SCID mice bearing OCI-AML3 xenografts
References	[1]. Highly effective combination of LSD1 (KDM1A) antagonist and pan-histone deacetylase inhibitor against human AML cells. Leukemia. 2014 Nov;28(11):2155-64. [2]. Stepwise assembly of functional C-terminal REST/NRSF transcriptional repressor complexes as a drug target. Protein Sci. 2017 Feb 20.

Solubility Data

Solubility (In Vitro)

DMSO: 38 mg/mL (86.8 mM) Water:<1 mg/mL Ethanol:<1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.75 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 20.8 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: 10% DMSO+corn oil: 5mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2836 mL	11.4181 mL	22.8363 mL
	5 mM	0.4567 mL	2.2836 mL	4.5673 mL
	10 mM	0.2284 mL	1.1418 mL	2.2836 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.