ASM-IN-2 (compound 46) is a potent and potent inhibitor of ASM with an IC50 value of 0.87 μM and exhibits good drug-like properties. ASM-IN-2 is involved in multiple antidepressant mechanisms, which are related to the decrease of ceramides, and exhibits significant antidepressant effects in CUMS-induced mouse models, and is expected to be used in research in the field of antidepressant drugs.

Physicochemical Properties

Molecular Formula	C16H11BRCLN3O2
Molecular Weight	392.63
CAS #	2305789-66-4
Appearance	Typically exists as solid at room temperature
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	PLC 0.87 μM (IC50)
ln Vitro	ASM-IN-2 showed the highest activity in binding to ASM, with an IC50 value of 0.87 μM[1]. In the anti-proliferation experiment of human neuroblastoma SH-SY5Y cells detected by CCK8, ASM-IN-2 showed slight cytotoxicity at a higher concentration of 100 μM (cell survival rate was higher than 78%)[1].
ln Vivo	ASM-IN-2 (1 or 10 MG/KG, iv or ip) is rapidly absorbed (Tmax = 0.25 h) in male ICR mice, reaching higher maximum concentrations (Cmax = 4204.13 NG/ML) and moderate biotoxicity Utilization (F = 39.59%), suitable for research on antidepressants [1]. ASM-IN-2 (6, 12, 24 MG/KG, ip, 10 days) significantly reduced the immobility time of mice in the forced swimming test (FST) and tail suspension test (TST), and the effect was better than that of fluoxan Ting (12 MG/KG) (HY-W011235). ASM-IN-2 significantly increases BDNF expression and the level of sphingolipid ceramide in the cerebral cortex, and also improves oxidative stress levels by inhibiting AMS activity [1]. ASM-IN-2 (6, 12, 24 MG/KG, ip) reduces the levels of TNF-α, IL-1β and IL-6 in a dose-dependent manner in the CUMS mouse model, attenuating the inflammatory response[1]. ASM-IN-2 (6, 12, 24 MG/KG, ip) significantly increases the 5-HT content in mouse hippocampus tissue, thereby promoting more effective antidepressant activity by inhibiting ASM [1].
Cell Assay	Cell Viability Assay[1] Cell Types: Corticosterone (CORT)-induced human neuroblastoma Tested Concentrations: 5, 10, 20 μM Incubation Duration: 24 h Experimental Results: Significantly attenuated corticosterone-induced damage in a dose-dependent manner and enhanced cell viability. Cell Cytotoxicity Assay[1] Cell Types: Human neuroblastoma cell line Concentration: 5, 10, 20, 50, 100 μM Incubation Duration: Experimental Results: Didn’t show any visible neurotoxicity at concentrations of 5, 10, and 20 μM, whereas exhibited weak cytotoxicity at a high concentration of 100 μM.
References	[1]. Discovery of a Novel ASM Direct Inhibitor with a 1,5-Diphenyl-pyrazole Scaffold and Its Antidepressant Mechanism of Action[J]. J Med Chem. 2024 Jun 18:1520-4804.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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	2.5469 mL	12.7346 mL	25.4693 mL
	5 mM	0.5094 mL	2.5469 mL	5.0939 mL
	10 mM	0.2547 mL	1.2735 mL	2.5469 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.