S2157 is an N-alkylated trans-cyclopropylamine (TCP) analogue and a potent lysine-specific demethylase 1 (LSD1) inhibitor. S2157 increases H3K9 methylation and corresponding H3K27 deacetylation in super-enhancer regions. S2157 inhibits the transcription of NOTCH3 and TAL1 genes, thereby inducing apoptosis in TCP-resistant acute lymphoblastic leukemia T-ALL cells. S2157 effectively penetrates the BBB (blood-brain barrier) and can almost completely eradicate central nervous system/CNS leukemia in mice transplanted with T-ALL cells.

Physicochemical Properties

Molecular Formula	C23H28CLF2N3O2
Molecular Weight	451.94
Exact Mass	451.183
CAS #	2262488-39-9
PubChem CID	141709235
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	7
Heavy Atom Count	31
Complexity	567
Defined Atom Stereocenter Count	2
SMILES	N([C@@H]1C[C@H]1C1C=C(F)C=C(F)C=1OCC1C=CC=CC=1)CC(N1CCN(C)CC1)=O.Cl
InChi Key	PUXYOFXCUCMFSQ-OZYANKIXSA-N
InChi Code	InChI=1S/C23H27F2N3O2.ClH/c1-27-7-9-28(10-8-27)22(29)14-26-21-13-18(21)19-11-17(24)12-20(25)23(19)30-15-16-5-3-2-4-6-16;/h2-6,11-12,18,21,26H,7-10,13-15H2,1H3;1H/t18-,21+;/m0./s1
Chemical Name	2-[[(1R,2S)-2-(3,5-difluoro-2-phenylmethoxyphenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)ethanone;hydrochloride
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	LSD1[1]
ln Vitro	S2157 is especially useful for T-ALL cell lines, with IC50 values ranging from 6.8 µM for MOLT4 to 1.1 µM for human T-ALL cell lines CEM[1]. S2157 (4–20 µM; 72 hours) slightly inhibits normal T-cells stimulated by mitogens[1]. In T-cell acute lymphoblastic leukemia (T-ALL) cells, S2157 (4–8 µM; 24 hours) causes apoptosis and down-regulates the expression of NOTCH3 and TAL1 proteins[1].
ln Vivo	Subcutaneous tumors treated with S2157 (50 mg/kg; IP; three times a week; for 28 days) show a reduction in size to less than 20% of the untreated control[1]. The drug S2157 (50 mg/kg; IP) possesses an AUC of 5.75 μM·h, a Cmax of 4.33 μM, and a T1/2 of 0.88 hours[1]. In the majority of NOD/SCID mice containing MOLT4 cells, but not all of them, S2157 (30 mg/kg or 50 mg/kg; twice weekly for three weeks) nearly entirely inhibited the proliferation of MOLT4 cells. Murine xenotransplanted models of CNS leukemia are eradicated by S2157[1].
Cell Assay	Cell Viability Assay[1] Cell Types: Normal T-lymphocytes Tested Concentrations: 4, 8, 12, 16, 20 µM Incubation Duration: For 72 hrs (hours) Experimental Results: Modestly inhibited mitogen-activated normal T-lymphocytes. Apoptosis Analysis[1] Cell Types: T-cell acute lymphoblastic leukemia (T-ALL) cells Tested Concentrations: 4, 6, 8 µM Incubation Duration: For 24 hrs (hours) Experimental Results: Induced apoptosis, as evidenced by increased annexin-V reactivity on flow cytometry in T-ALL cells in a dose- and time-dependent manner without affecting cell cycle distribution. Western Blot Analysis[1] Cell Types: T-ALL cells Tested Concentrations: 4, 6, 8 µM Incubation Duration: For 24 hrs (hours) Experimental Results: Down-regulated the expression of NOTCH3 and TAL1 proteins in T-ALL cells.
Animal Protocol	Animal/Disease Models: Nonobese diabetic/ severe combined immunodeficiency (NOD/SCID) mice with MOLT4 cells[1] Doses: 50 mg/kg Route of Administration: IP; 3 times a week; for 28 days Experimental Results: The size of subcutaneous (sc)tumors diminished to less than 20% of that in the untreated control. Animal/Disease Models: 8weeks old ICR mice[1] Doses: 50 mg/kg (pharmacokinetic/PK Analysis) Route of Administration: IP Experimental Results: Had a T1/2 of 0.88 hrs (hours), a Cmax of 4.33 μM and an AUC of 5.75 μM·h.
References	[1]. Eradication of Central Nervous System Leukemia of T-Cell Origin With a Brain-Permeable LSD1 Inhibitor. Clin Cancer Res. 2019 Mar 1;25(5):1601-1611.

Solubility Data

Solubility (In Vitro)

DMSO : 100 mg/mL (221.27 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	2.2127 mL	11.0634 mL	22.1268 mL
	5 mM	0.4425 mL	2.2127 mL	4.4254 mL
	10 mM	0.2213 mL	1.1063 mL	2.2127 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.