Encukalner specifically targets Kv7.2/Kv7.3 (KCNQ2/3) potassium channels. It acts as a selective opener (activator) of these channels with an EC50 of 27 nM. It exhibits approximately four-fold selectivity for the Kv7.2/7.3 combination over Kv7.3/7.5 and Kv7.4 channels, with EC50 values of 94 nM and 113 nM, respectively.

In vitro, Encukalner significantly increases potassium currents in cells expressing KCNQ2/3 channels. It demonstrates potent channel-opening activity with an EC50 of 27 nM. The compound shows selectivity for Kv7.2/7.3 heteromeric channels over other Kv7 channel subtypes, making it a valuable tool for studying the physiology of these channels.

In vivo, Encukalner shows potent anticonvulsant activity in multiple rodent seizure models, including the maximal electroshock seizure (MES) and pentylenetetrazol (PTZ) models. It has also been investigated for its potential in treating pain. The compound's oral activity makes it suitable for chronic administration in preclinical efficacy studies.

The activity of Encukalner on Kv7.2/Kv7.3 channels is assessed using electrophysiological techniques, particularly patch-clamp recordings. HEK293 cells expressing recombinant KCNQ2/3 channels are treated with varying concentrations of the compound. The increase in potassium current amplitude is measured. EC50 values are calculated from concentration-response curves.

Cellular assays for Encukalner involve the use of cell lines expressing recombinant human KCNQ2/3 potassium channels. The compound's effect on membrane potential is measured using fluorescent membrane potential-sensitive dyes (e.g., DiBAC₄(3)) or FLIPR-based assays. The compound's ability to enhance potassium currents is confirmed by patch-clamp electrophysiology.

In vivo studies of Encukalner are conducted in rodent models of epilepsy and pain. For seizure models, the compound is administered orally or intraperitoneally prior to seizure induction using MES or PTZ. The latency to seizure onset, seizure severity, and mortality are recorded. For pain models, the compound's analgesic effects are assessed using thermal or mechanical nociception tests.

Pharmacokinetic properties of Encukalner are evaluated in rodents and other species following oral administration. The compound is formulated in suitable vehicles (e.g., 0.5% methylcellulose). Plasma and brain concentrations are measured using LC-MS/MS to determine key PK parameters including oral bioavailability, half-life, and brain penetration.

Encukalner is an investigational drug that has been evaluated in clinical trials for epilepsy and major depressive disorder. Standard toxicology assessments have been conducted as part of its preclinical development program. The compound is generally well-tolerated in preclinical species, though specific toxicity data is not detailed in public sources.

1: Yang GM, Tian FY, Shen YW, Yang CY, Yuan H, Li P, Gao ZB. Functional characterization and in vitro pharmacological rescue of KCNQ2 pore mutations associated with epileptic encephalopathy. Acta Pharmacol Sin. 2023 Mar 17. doi: 10.1038/s41401-023-01073-y. Epub ahead of print. PMID: 36932231. 2: Elkommos S, Mula M. Current and future pharmacotherapy options for drug- resistant epilepsy. Expert Opin Pharmacother. 2022 Dec;23(18):2023-2034. doi: 10.1080/14656566.2022.2128670. Epub 2022 Sep 27. PMID: 36154780. 3: Bialer M, Johannessen SI, Koepp MJ, Levy RH, Perucca E, Perucca P, Tomson T, White HS. Progress report on new antiepileptic drugs: A summary of the Sixteenth Eilat Conference on New Antiepileptic Drugs and Devices (EILAT XVI): II. Drugs in more advanced clinical development. Epilepsia. 2022 Nov;63(11):2883-2910. doi: 10.1111/epi.17376. Epub 2022 Aug 11. PMID: 35950617. 4: Biondi A, Rocchi L, Santoro V, Rossini PG, Beatch GN, Richardson MP, Premoli I. Spontaneous and TMS-related EEG changes as new biomarkers to measure anti- epileptic drug effects. Sci Rep. 2022 Feb 4;12(1):1919. doi: 10.1038/s41598-022-05179-x. PMID: 35121751; PMCID: PMC8817040. 5: Bialer M, Johannessen SI, Koepp MJ, Levy RH, Perucca E, Perucca P, Tomson T, White HS. Progress report on new antiepileptic drugs: A summary of the Fifteenth Eilat Conference on New Antiepileptic Drugs and Devices (EILAT XV). I. Drugs in preclinical and early clinical development. Epilepsia. 2020 Nov;61(11):2340-2364. doi: 10.1111/epi.16725. Epub 2020 Nov 14. PMID: 33190243. 6: Premoli I, Rossini PG, Goldberg PY, Posadas K, Green L, Yogo N, Pimstone S, Abela E, Beatch GN, Richardson MP. TMS as a pharmacodynamic indicator of cortical activity of a novel anti-epileptic drug, XEN1101. Ann Clin Transl Neurol. 2019 Nov;6(11):2164-2174. doi: 10.1002/acn3.50896. Epub 2019 Sep 30. PMID: 31568714; PMCID: PMC6856596. 7: Bialer M, Johannessen SI, Koepp MJ, Levy RH, Perucca E, Tomson T, White HS. Progress report on new antiepileptic drugs: A summary of the Fourteenth Eilat Conference on New Antiepileptic Drugs and Devices (EILAT XIV). I. Drugs in preclinical and early clinical development. Epilepsia. 2018 Oct;59(10):1811-1841. doi: 10.1111/epi.14557. PMID: 30368792.

Encukalner (XEN1101) is an investigational drug that has advanced to clinical trials. It has been studied in Phase 2 clinical trials for refractory focal onset seizures and major depressive disorder. The compound's mechanism of action—opening Kv7.2/Kv7.3 potassium channels—stabilizes neuronal membrane potential and reduces neuronal excitability, providing a rationale for its anticonvulsant and antidepressant effects.

C23H29FN2O

368.5

368.226

C, 74.97; H, 7.93; F, 5.16; N, 7.60; O, 4.34

1009344-33-5

24743936

Off-white to gray solid powder

1.1±0.1 g/cm3

541.8±50.0 °C at 760 mmHg

281.4±30.1 °C

0.0±1.4 mmHg at 25°C

1.579

5.28

1

3

4

27

505

0

FC1C=CC2=C(C=1)CCN(C1C=C(C)C(=C(C)C=1)NC(CC(C)(C)C)=O)C2

FJNPZKZPWVVSON-UHFFFAOYSA-N

InChI=1S/C23H29FN2O/c1-15-10-20(11-16(2)22(15)25-21(27)13-23(3,4)5)26-9-8-17-12-19(24)7-6-18(17)14-26/h6-7,10-12H,8-9,13-14H2,1-5H3,(H,25,27)

N-[4-(6-fluoro-3,4-dihydroisoquinolin-2(1H)-yl)-2,6-dimethylphenyl]-3,3-dimethylbutanamide

Encukalner; XEN1101; XEN-1101 XEN 1101; 1OP 2198; VRX 621698; XPF-008;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~50 mg/mL (~135.69 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.78 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.

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 (Please use freshly prepared in vivo formulations for optimal results.)