Apinocaltamide (ACT-709478) is an orally bioavailable, BBB (blood-brain barrier) permeable/penetrable, selective T-type voltage-gated calcium channel (T-type calcium channel) blocker that may be utilized in the research/study of generalized epilepsy.

Physicochemical Properties

Molecular Formula	C22H18F3N5O
Molecular Weight	425.40643453598
Exact Mass	425.146
CAS #	1838651-58-3
PubChem CID	118560618
Appearance	White to off-white solid powder
LogP	3.3
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	31
Complexity	693
Defined Atom Stereocenter Count	0
InChi Key	LSYANGLAZUZYFX-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H18F3N5O/c23-22(24,25)21(8-9-21)17-4-1-15(2-5-17)11-20(31)28-19-7-10-30(29-19)14-18-6-3-16(12-26)13-27-18/h1-7,10,13H,8-9,11,14H2,(H,28,29,31)
Chemical Name	N-[1-[(5-cyanopyridin-2-yl)methyl]pyrazol-3-yl]-2-[4-[1-(trifluoromethyl)cyclopropyl]phenyl]acetamide
Synonyms	ACT709478 ACT 709478ACT-709478
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	Cav3.1, Cav3.2, Cav3.3, and Cav1.2 are all blocked by apinocaltamide (Compound 66b), with IC50 values of 6.4, 18, 7.5, and 2410 nM, respectively. The recombinant channel hCav3.3 is efficiently blocked by apinocaltamide with a notable voltage dependence (Kr≈1500 nM and Ki≈20 nM). With an IC50 of 5.5 μM, apinocaltamide inhibits current flow across hKv11.1-hERG channels [1]. Moreover, apinocaltamide inhibits P450 enzymes; its IC50 values for CYP2C8, CYP2D6, CYP2C9, CYP2C19, CYP3A4, and CYP2B6 are 14, 15, 22, 25, 51, and 52 μM, respectively [1].
ln Vivo	Apinocaltamide (66b, 100, and 300 mg/kg, orally administered, assessed 12 hours later) efficiently reduces the total amount of time mice experience absence-like seizures [1].
Animal Protocol	Animal/Disease Models: Male juvenile DBA/2J mice (22-24 days old) [1] Doses: 100, 300 mg/kg, 1 hour or 3 hrs (hrs (hours)) before exposure to stimulation. Route of Administration: Orally for 12 hrs (hrs (hours)) Experimental Results: Cumulative duration of absence seizures was diminished by 93% over the next 12 hrs (hrs (hours)).
References	[1]. Discovery of a Potent, Selective T-type Calcium Channel Blocker as a Drug Candidate for the Treatment of Generalized Epilepsies. J Med Chem. 2017 Dec 14;60(23):9769-9789.
Additional Infomation	Apinocaltamide is a secondary carboxamide resulting from the formal condensation of the carboxy group of {4-[1-(trifluoromethyl)cyclopropyl]phenyl}acetic acid with the amino group of 6-[(3-amino-1H-pyrazol-1-yl)methyl]pyridine-3-carbonitrile. It is a selective, orally available T-type calcium channel blocker that is being studied as a potential new treatment in epilepsy. It has a role as an anticonvulsant and a T-type calcium channel blocker. It is a member of pyrazoles, a secondary carboxamide, a member of cyclopropanes, an organofluorine compound, a nitrile, a member of pyridines and a member of benzenes. Apinocaltamide is under investigation in clinical trial NCT03239691 (A Study to Evaluate the Effect of ACT-709478 in Photosensitive Epilepsy Patients).

Solubility Data

Solubility (In Vitro)

DMSO : ≥ 125 mg/mL (~293.83 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.89 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: ≥ 2.08 mg/mL (4.89 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 20.8 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: ≥ 2.08 mg/mL (4.89 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 20.8 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	2.3507 mL	11.7534 mL	23.5067 mL
	5 mM	0.4701 mL	2.3507 mL	4.7013 mL
	10 mM	0.2351 mL	1.1753 mL	2.3507 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.