ML380 is a potent, isoform-selective and BBB (blood-brain barrier) permeable/penetrable M5 mAChR positive allosteric modulator (PAM) with EC50s for human and rat M5 of 190 and 610 nM, respectively. ML380 is moderately selective for the M1 and M3 mAChR subtypes. ML380 can increase the affinity of ACh for M5 mAChR.

Physicochemical Properties

Molecular Formula	C23H25F3N4O3S
Molecular Weight	494.53
Exact Mass	494.159
CAS #	1627138-52-6
PubChem CID	71737672
Appearance	White to off-white solid powder
Density	1.4±0.1 g/cm3
Boiling Point	661.2±65.0 °C at 760 mmHg
Flash Point	353.7±34.3 °C
Vapour Pressure	0.0±2.0 mmHg at 25°C
Index of Refraction	1.601
LogP	3.52
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	6
Heavy Atom Count	34
Complexity	811
Defined Atom Stereocenter Count	0
InChi Key	FJPQEOQGIZSLES-UHFFFAOYSA-N
InChi Code	InChI=1S/C23H25F3N4O3S/c1-2-29(15-17-5-3-4-6-20(17)23(24,25)26)22(31)16-9-11-30(12-10-16)34(32,33)19-7-8-21-18(13-19)14-27-28-21/h3-8,13-14,16H,2,9-12,15H2,1H3,(H,27,28)
Chemical Name	N-ethyl-1-(1H-indazol-5-ylsulfonyl)-N-[[2-(trifluoromethyl)phenyl]methyl]piperidine-4-carboxamide
Synonyms	ML-380; ML 380; ML380
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

ln Vitro	In CHO-hM5 cells, ML380 (0.01 nM-100 μM) significantly increases the accumulation of inositol phosphate (IP) and Ca2+ mobilization, with pEC50 values of 5.33 and 5.71, respectively [2]. ML380 (0.01-30 μM) stimulates IP accumulation and Ca2+ mobilization in CHO-hM5 cells in response to ACh [2].
ln Vivo	In rats, ML380 (1 mg/kg; iv) has a short half-life (t1/2, 22 min), a moderate volume of distribution (1.6 L/kg), and a high clearance (66 mL/min/kg). 1].
References	[1]. Development of a highly potent, novel M5 positive allosteric modulator (PAM) demonstrating CNS exposure: 1-((1H-indazol-5-yl)sulfoneyl)-N-ethyl-N-(2-(trifluoromethyl)benzyl)piperidine-4-carboxamide (ML380). J Med Chem. 2014 Sep 25;57(18. [2]. Molecular Mechanisms of Action of M5 Muscarinic Acetylcholine Receptor Allosteric Modulators. Mol Pharmacol. 2016 Oct;90(4):427-36. [3]. Structure-Activity Relationships of Pan-Gα q/11 Coupled Muscarinic Acetylcholine Receptor Positive Allosteric Modulators. ACS Chem Neurosci. 2018 Jul 18;9(7):1818-1828.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~202.21 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.06 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 (5.06 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 25.0 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.06 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	2.0221 mL	10.1106 mL	20.2212 mL
	5 mM	0.4044 mL	2.0221 mL	4.0442 mL
	10 mM	0.2022 mL	1.0111 mL	2.0221 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.