CA3042004C

CA3042004C - Benzenesulfonamide compounds and their use as therapeutic agents

Info

Publication number: CA3042004C
Application number: CA3042004A
Authority: CA
Inventors: David Earl Bogucki; Sultan Chowdhury; Qi Jia; Alla Yurevna Zenova; Jean-Christophe Andrez; Michael Scott Wilson; Christoph Martin Dehnhardt; Thilo Focken; Michael Edward GRIMWOOD; Shannon Marie Decker; Kristen Nicole Burford; Charles Jay Cohen; Robert Joseph Devita; James Roy Empfield; Syed Abid Hasan; Jr. Johnson James Philip
Original assignee: Xenon Pharmaceuticals Inc
Priority date: 2016-12-09
Filing date: 2017-05-19
Publication date: 2025-10-07
Anticipated expiration: 2037-05-19

Abstract

This invention is directed to benzenesulfonamide compounds, as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof, for the treatment of diseases or conditions associated with voltage-gated sodium channels, such as epilepsy.

Description

DEMANDE OU BREVET VOLUMINEUX LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND PLUS D’UN TOME. CECI EST LE TOME 3 DE 3 CONTENANT LES PAGES 572 A 589 NOTE : Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE VOLUME THIS IS VOLUME 3 OF 3 CONTAINING PAGES 572 TO 589 NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME : NOTE POUR LE TOME / VOLUME NOTE:5 10 15 20 25 30 CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Following the procedure as described in EXAMPLE 145, step 5 and making non-critical variations to replace tert-butyl (2-(azetidin-1-ylmethyl)-6-fluorobenzyl)(4-(/V- (2,4-dimethoxybenzyl)-/V-(pyrimidin-2-yl)sulfamoyl)-3,5-difluorophenyl)carbamate with tert-butyl (S)-((5-chloro-2-fluoro-4-(1-(2-fluorophenyl)propoxy)phenyl)sulfonyl)(thiazol- 4-yl)carbamate, and purification preparative reverse phase HPLC, using a gradient of acetonitrile in water (containing 0.225% of formic acid), the title compound was afforded as a colorless solid (0.069 g, 30% yield): 1H NMR (400 MHz, CDCI3) <59.33 (s, 1H), 8.63 (d, J = 2.4 Hz, 1H), 7.87 (d, J = 7.6 Hz, 1H), 7.34-7.30 (m, 2H), 7.14 (d, J = 7.6 Hz, 2H), 6.98 (d, J = 2.0 Hz, 1H), 6.54 (d, J= 11.6 Hz, 1H), 5.46 (t, J=6.4 Hz, 1H), 2.16 (q, J= 7.2 Hz, 1H), 2.01 (q, J = 7.2 Hz, 1H), 1.06 (t, J= 7.2 Hz, 3H); MS (ES+) m/z 445.1 (M + 1). BIOLOGICAL ASSAYS Various techniques are known in the art for testing the activity of the compound of the invention or determining their solubility in known pharmaceutically acceptable excipients. In order that the invention described herein may be more fully understood, the following biological assays are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any manner. BIOLOGICAL EXAMPLE 1 Electrophysiological Assay (In vitro assay) Patch voltage clamp electrophysiology allows for the direct measurement and quantification of block of voltage-gated sodium channels (Nay’s), and allows the determination of the time- and voltage-dependence of block which has been interpreted as differential binding to the resting, open, and inactivated states of the sodium channel (Hille, B., Journal of General Physiology (1977), 69: 497-515). The following patch voltage clamp electrophysiology studies were performed on representative compounds of the invention using human embryonic kidney cells (HEK), permanently transfected with an expression vector containing the full-length cDNA coding for the desired human sodium channel a-subunit, grown in culture media 5695 10 15 20 25 30 Date Re^ue/Date Received 2023-10-18 CA 3042004 containing 10% FBS, 1% PSG, and 0.5 mg/mL G418 at 37 °C with 5% CO2. HEK cells used for the electrophysiology (EP) recordings had a passage number of less than 40 for all studies and were used within three days from the time of plating. Na/1.1, Nad.5 and Nad.6 cDNAs (NM_001165964 (SCN1A), NM_000335 (SCN5A) and NM_014191 (SCN8A), respectively) were stably expressed in HEK-293 cells. Sodium currents were measured using the patch clamp technique in the whole-cell configuration using either a PatchXpress automated voltage clamp or manually using an Axopatch™ 200B (Axon Instruments) or Model 2400 (A-M systems) amplifier. The manual voltage clamp protocol was as follows: Borosilicate glass micropipettes were fire-polished to a tip diameter yielding a resistance of 2-4 Mohms in the working solutions. The pipette was filled with a solution comprised of: 5 mM NaCI, 10 mM CsCI, 120 mM CsF, 0.1 mM CaCh, 2 mM MgCh, 10 mM HEPES, 10 mM EGTA; and adjusted to pH 7.2 with CsOH. The external solution had the following composition: 140 mM NaCI, 5 mM KCI, 2 mM CaCh, 1 mM MgCh, 10 mM HEPES; and adjusted to pH 7.4 with NaOH. In some studies, the external sodium was reduced by equimolar replacement with choline. Osmolarity in the CsF internal and NaCI external solutions was adjusted to 300 mOsm/kg and 310 mOsm/kg with glucose, respectively. All recordings were performed at ambient temperature in a bath chamber with a volume of 150 pL. Control sodium currents were measured in 0.5% DMSO. Controls and representative compounds of the invention were applied to the recording chamber through a 4-pinch or 8- pinch valve bath perfusion system manufactured by ALA Scientific Instruments. Currents were recorded at 40 kHz sampling frequency, filtered at 5 Hz, and stored using a DigidataTM-1322A analogue/digital interface with the pCIamp software (Axon Instruments). Series resistance compensation was applied (60-80%). Cells were rejected if currents showed inadequate voltage control (as judged by the IV relationship during stepwise activation). All statistics in this study are given as mean ± SD. The membrane potential was maintained at a voltage where inactivation of the channel is complete. The voltage is then stepped back to a very negative (Vhold = -150mV) voltage for 20 ms and then a test pulse is applied to quantify the compound block. The 20 ms brief repolarization was long enough for compound-free channels to completely recover from fast inactivation, but the compound-bound channels recovered more slowly such that negligible recovery could occur during this interval. The percent decrease in sodium current following wash-on of compound was taken as the percent block of sodium channels. 5705 10 15 20 25 Date Re^ue/Date Received 2023-10-18 CA 3042004 Representative compounds of the invention, when tested in this assay, demonstrated the ICso's as set forth below in Table 1. BIOLOGICAL EXAMPLE 2 Sodium Influx Assay (In vitro assay) This sodium influx assay employs the use of the cell permeable, sodium sensitive dye ANG2 to quantify sodium ion influx through sodium channels which are maintained in an open state by use of sodium channel modulators. This high throughput sodium influx assay allows for rapid profiling and characterization of sodium channel blockers. In general, Trex™ HEK293 cells were stably transfected with an inducible expression vector containing the full-length cDNA coding for the desired human sodium channel a-subunit and with an expression vector containing full length cDNA coding for the |31-subunit. Sodium channel expressing cell lines were induced with tetracycline (Ipg/mL) and plated on 384-well PDL-coated plates at a density of 25K-30K cells/ well in culture media (DMEM, containing 10% FBS and 1% L-glutamine). After overnight incubation (37 °C, 5% CO2), culture media was removed and cells were loaded with 5uM ANG2 dye for 1-1.5h in Buffer 1 (155 mM NMDG, 5 mM KCI, 2 mM CaCh, 1 mM MgCh, 10 mM HEPES, 10 mM glucose, adjusted with Tris to pH 7.4). Access dye was removed and cells were incubated with test compounds for 1hr in buffer 1 containing sodium channel modulator(s) at room temperature. Hamamatsu FDSS pCell was used to perform a 1:1 addition of Na/K challenge buffer (140 mM NaCI, 20 mM HEPES, 1 mM CaCfe, 15 mM KCI, 1 mM MgCh, 10 mM glucose, adjusted with Tris to pH 7.4) and simultaneously read plates at excitation wavelength of 530 nm and emission wavelength set at 558 nm. Percent inhibition of sodium ion influx was calculated for each test compound at each test concentration to determine the IC50 values. Representative compounds of the invention, when tested in this model, demonstrated affinities for the inactivated state of Na/1.6, Na/1.5 and Na/1.1 as set forth below in Table 1. The Example numbers provided in Table 1 correspond to the Examples herein, "Flux" refers to the Sodium Influx Assay and "EP" refers to the Electrophysiological Assay : 571CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 TABLE 1: Inhibition of Navi.1, Nav1.5, and Na/I.6 Ex. No. Flux Na/I.6 IC50(pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EPNav1.5 IC50 (UM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 1 0.120 8.916 0.784 2 5.021 8.246 15.982 3 0.679 5.254 3.379 4 0.794 0.041 7.010 3.401 5.306 5 0.785 0.152 5.311 10.000 4.831 10.000 6 3.377 5.516 3.758 7 0.376 9.682 2.995 8 0.137 0.010 5.366 1.626 0.420 9 0.181 0.026 4.222 2.363 4.335 10 0.461 0.128 2.349 1.630 5.474 11 0.255 0.034 30.000 30.000 10.000 12 1.980 30.000 30.000 13 5.888 30.000 30.000 14 1.790 14.431 4.651 15 7.149 23.642 9.409 16 0.029 0.004 23.462 5.408 2.189 17 0.308 8.459 0.494 18 3.351 11.888 8.373 19 0.425 0.130 18.511 11.669 10.000 20 2.052 1.177 30.000 30.000 10.000 21 3.668 2.888 3.631 22 1.296 0.259 4 855 4.291 9.000 23 1.519 4.088 5.398 24 4.092 8.325 5.851 25 0.175 0.057 10.297 1.872 2.516 572CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 26 0.742 0.113 3.452 4.742 6.751 27 0.069 0.012 8.493 0.476 0.112 28 0.876 0.406 8.254 7.367 10.000 29 30.000 24.596 30.000 30 1.441 0.652 12.661 10.848 10.000 31 0.917 0.118 16.934 6.168 10.000 32 0.092 0.023 12.788 1.056 33 0.462 25.817 2.992 34 1.271 4.910 0.840 35 0.839 8.691 0.301 36 1.398 5.117 1.341 37 2.088 12.346 1.904 38 0.341 0.024 9.892 10.000 2.645 0.994 39 0.791 0.285 6.806 2.350 1.846 40 0.123 0.024 7.898 0.637 0.182 41 0.070 0.010 11.777 0.364 0.031 42 3.026 0.305 14.441 5.646 6.768 43 0.191 0.028 11.122 2.139 4.103 44 0.347 0.130 30.000 1.271 2.091 45 0.108 0.013 6.259 5.151 1.176 0.539 46 1.662 7.040 0.999 47 0.089 0.010 6.515 0.816 0.114 48 0.716 0.230 8.656 1.358 2.604 49 0.232 0.019 4.746 0.888 0.260 50 0.169 6.073 1.278 51 10.850 20.603 4.274 573CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 52 0.234 0.037 10.482 1.545 0.590 53 0.741 6.889 0.726 54 0.265 0.025 13.516 3.244 1.025 55 20.715 21.253 16.083 56 0.949 0.292 30.000 4.031 1.415 57 2.293 30.000 2.599 58 0.152 7.086 2.529 59 0.387 0.077 30.000 8.394 1.718 60 3.341 30.000 1.570 61 0.021 0.004 8.910 10.000 1.129 0.283 62 0.138 0.052 30.000 17.251 3.098 63 0.392 0.066 30.000 14.147 10.000 64 0.197 0.085 30.000 14.476 10.000 65 1.019 0.087 30.000 2.230 10.000 66 1.012 1.113 30.000 0.908 10.000 67 0.426 0.119 30.000 10.568 10.000 68 1.808 25.284 5.148 69 1.591 7.662 3.030 70 0.181 0.036 30.000 30.000 10.000 71 30.000 30.000 30.000 72 7.105 0.955 30.000 30.000 10.000 73 3.868 1.139 30.000 23.119 10.000 74 0.521 0.052 30.000 2.518 1.853 75 0.261 21.671 13.646 76 0.397 0.126 10.276 3.971 10.000 77 8.079 26.085 7.975 574CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 78 0.234 0.015 7.008 4.169 3.103 79 0.237 0.039 2.548 1.542 4.406 80 1.744 0.099 5.752 11.137 10.000 81 0.652 0.143 30.000 22.468 10.000 82 2.155 30.000 28.736 83 12.338 30.000 11.013 84 1.169 1.773 19.337 2.413 10.000 85 0.107 0.078 30.000 0.516 86 4.060 0.558 12.743 16.643 87 8.596 24.672 30.000 88 1.041 0.184 18.215 30.000 10.000 89 0.948 0.239 9.090 3.851 10.000 90 3.062 1.411 0.969 3.822 4.046 2.012 91 0.927 0.341 30.000 10.672 10.000 92 2.533 4.431 7.454 93 0.270 8.468 0.314 94 0.423 8.620 0.142 95 0.761 30.000 8.112 96 0.239 0.098 15.981 12.647 10.000 97 2.974 30.000 4.966 98 0.299 11.024 0.303 99 0.373 9.270 1.938 100 0.216 6.420 0.667 101 0.814 0.462 4.655 9.568 10.000 102 1.667 5.971 0.941 103 2.906 0.796 28.788 16.616 10.000 575CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 104 0.582 0.153 6.285 4.432 4.510 105 0.564 0.027 10.564 3.727 3.598 106 0.533 10.140 3.701 107 0.656 0.138 7.403 10.295 8.617 108 14.534 22.353 21.084 109 0.214 7.861 1.109 110 1.030 3.152 3.837 111 0.352 8.334 1.385 112 0.821 16.079 3.108 113 0.156 0.044 25.662 1.886 7.967 114 0.977 0.255 4.638 5.054 10.000 115 1.388 0.135 9.926 17.527 10.000 116 0.477 0.037 4.580 14.280 5.775 9.891 117 0.530 0.055 6.216 5.892 6.782 118 3.181 7.502 15.944 119 0.310 0.023 1.825 5.719 8.942 120 1.392 0.717 2.416 24.283 10.000 121 2.324 2.154 3.378 122 1.654 0.121 30.000 30.000 10.000 123 1.169 0.716 2.147 5.784 10.000 124 1.842 1.871 15.897 125 1.733 3.353 3.257 126 0.114 11.443 0.147 127 0.515 0.229 30.000 30.000 10.000 128 0.208 6.715 5.368 129 0.322 0.252 30.000 4.244 10.000 576CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 130 0.307 0.100 17.669 11.662 10.000 131 0.376 0.126 10.928 8.328 10.000 132 5.108 24.684 7.868 133 0.418 0.122 9.210 6.357 10.000 134 0.643 15.038 10.786 135 0.444 22.826 7.100 136 0.384 0.041 23.699 19.574 10.000 137 0.588 0.353 30.000 30.000 10.000 138 0.151 0.043 9.847 5.758 10.000 139 0.107 0.038 12.879 30.000 1.292 140 0.585 0.050 12.498 5.562 141 0.479 0.031 21.015 3.331 0.277 142 0.187 0.106 21.247 3.227 10.000 143 0.528 0.027 7.011 2.492 0.159 144 0.214 0.034 9.674 2.545 0.907 145 0.467 30.000 30.000 146 4.122 9.844 5.574 147 3.310 2.723 3.430 148 1.253 4.837 2.954 149 0.780 6.386 3.358 150 2.819 8.657 30.000 151 2.095 30.000 20.505 152 0.160 18.980 10.284 153 0.016 30.000 17.465 154 0.451 30.000 28.833 155 0.250 30.000 30.000 577CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 156 0.146 0.022 30.000 24.768 10.000 157 2.358 30.000 30.000 158 0.126 30.000 30.000 159 0.153 11.806 9.213 160 0.064 23.003 6.514 161 0.267 30.000 9.989 162 0.461 11.035 30.000 163 0.288 30.000 6.349 164 0.084 20.313 22.789 165 0.361 20.768 14.936 166 0.015 15.174 12.688 167 0.405 30.000 30.000 168 0.120 21.667 17.736 169 0.011 29.045 6.377 170 0.176 15.153 24.693 171 0.385 16.752 21.859 172 0.078 0.023 30.000 25.815 173 0.050 19.519 7.221 174 0.035 30.000 30.000 175 0.342 14.540 10.709 176 0.680 16.876 5.747 177 0.046 22.283 5.396 178 0.265 30.000 13.433 179 0.080 6.421 4.596 180 0.166 24.246 22.075 181 2.965 30.000 10.586 578CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 182 0.492 22.760 11.922 183 0.036 27.964 30.000 184 0.332 30.000 30.000 185 0.092 30.000 30.000 186 0.019 9.119 15.548 187 0.065 30.000 12.353 188 0.025 0.012 30.000 9.200 189 0.522 30.000 30.000 190 0.571 0.055 25.833 30.000 13.360 9.705 191 0.244 0.112 20.003 18.160 10.000 192 0.072 0.015 8.511 16.180 10.000 193 7.488 26.713 8.853 194 0.227 0.043 30.000 8.539 10.000 195 0.688 30.000 27.223 196 0.187 30.000 25.903 197 0.023 27.621 11.951 198 0.391 30.000 25.843 199 0.213 30.000 4.077 200 0.496 15.362 4.428 201 0.561 30.000 30.000 202 0.534 30.000 30.000 203 0.225 28.282 26.596 204 0.187 0.081 23.107 30.000 205 0.650 0.064 22.345 3.638 10.000 206 0.448 0.076 30.000 7.048 10.000 207 0.413 0.121 9.573 4.376 8.127 579CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 208 0.593 30.000 9.892 209 0.317 0.076 30.000 27.304 10.000 210 0.416 26.841 3.748 211 1.757 19.247 3.380 212 0.226 0.041 30.000 10.000 30.000 10.000 213 0.156 0.021 30.000 30.000 214 0.326 16.170 13.362 215 0.021 0.003 21.538 10.337 216 3.120 30.000 30.000 217 0.701 11.573 15.301 218 3.207 20 885 28.398 219 0.280 30.000 30.000 220 0.193 3.170 2.115 221 4.663 19.032 25.374 222 1.925 8.350 3.987 223 4.228 12.217 3.736 224 5.074 30.000 7.757 225 2.000 0.742 4.473 9.823 10.000 226 8.188 11.318 19.865 227 1.246 8.602 8.778 228 1.944 27.494 13.155 229 0.961 30.000 30.000 230 0.909 28.707 10.727 231 4.215 30.000 17.500 232 1.750 11 221 21.169 233 0.787 12.433 3.084 580CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 234 0.669 8.666 4.615 235 3.018 30.000 22.352 236 3.015 30.000 30.000 237 3.535 30.000 12.523 238 0.354 30.000 6.448 239 1.654 28.886 27.045 240 3.172 30.000 30.000 241 2.363 30.000 26.793 242 1.947 30.000 30.000 243 8.679 6.249 30.000 30.000 10.000 244 4.277 22.155 30.000 245 5.383 6.555 10.471 246 0.200 0.033 30.000 30.000 10.000 247 6.162 0.409 30.000 25.313 10.000 248 0.431 26.490 21.442 249 0.163 30.000 30.000 250 9.058 30.000 23.588 251 3.711 3.348 8.678 252 0.036 12.569 9.965 253 2.099 13.101 19.999 254 9.438 3.617 14.070 255 0.471 5.980 7.786 256 1.219 6.972 8.817 257 0.490 8.386 6.101 258 3.694 30.000 30.000 259 1.369 30.000 29.247 581CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 260 1.877 30.000 30.000 261 0.146 26.323 28.876 262 1.423 7.321 8.292 263 1.025 4.781 7.711 264 30.000 30.000 30.000 265 0.194 0.018 12.506 0.794 0.050 266 0.117 0.015 5.440 0.767 0.133 267 0.959 6.964 3.577 268 0.035 0.010 3.682 0.388 0.332 269 0.584 6.718 2.777 270 0.213 0.018 11.037 12.513 10.000 271 0.448 30.000 30.000 272 0.163 0.031 30.000 30.000 20.668 10.000 273 0.896 0.122 30.000 30.000 10.000 274 0.754 4.603 5.251 275 0.174 0.025 30.000 5.328 4.854 276 0.265 24.260 3.604 277 0.380 0.043 9.392 10.000 3.775 5.592 278 0.187 30.000 14.165 279 0.163 30.000 11.207 280 1.389 23.926 8.733 281 1.075 11.015 7.063 282 0.409 6.580 9.779 283 0.126 0.011 28.008 30.000 14.886 14.470 284 1.373 30.000 30.000 285 0.625 26.611 5.891 582CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 286 3.377 8.782 6.843 287 1.531 0.470 30.000 25.150 10.000 288 0.180 0.036 17.948 11.760 10.000 289 0.556 30.000 4.266 290 0.101 0.011 25.100 6.019 10.000 291 0.368 30.000 14.025 292 0.624 8.402 2.719 293 0.426 0.202 30.000 12.729 10.000 294 0.688 30.000 30.000 295 0.383 0.042 30.000 12.052 10.000 296 1.308 30.000 16.890 297 0.559 0.391 8.627 2.975 10.000 298 0.785 11.257 7.190 299 3.609 21.644 0.684 300 0.346 0.064 30.000 15.949 10.000 301 0.493 30.000 30.000 302 0.049 23.919 9.665 303 0.007 24.527 5.728 304 0.008 0.002 21.180 5.297 305 0.057 30.000 14.214 306 0.241 8.104 18.134 307 0.597 0.451 21.311 2.285 10.000 308 0.605 26.379 25.965 309 0.085 27.568 6.344 310 0.010 0.011 30.000 20.318 311 0.219 30.000 26.406 583CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/I.5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 312 0.135 9.590 7.256 313 0.014 14.313 3.746 314 0.032 30.000 30.000 315 0.270 30.000 30.000 316 0.961 0.264 5.864 2.497 5.654 317 0.328 0.982 30.000 3.907 318 6.026 17.287 20.408 319 2.308 20.016 18.015 320 0.055 17.906 6.505 321 0.091 28.138 4.770 322 0.114 0.047 30.000 7.753 323 0.518 15.037 9.380 324 0.647 30.000 18.892 325 0.401 30.000 30.000 326 0.125 0.090 12.727 7.864 10.000 327 0.003 0.001 30.000 4.443 328 3.818 1.039 30.000 30.000 329 0.214 28.413 8.116 330 0.001 0.001 30.000 1.996 331 0.724 0.099 11.269 7.883 10.000 332 0.275 7.239 0.831 333 0.257 0.186 30.000 30.000 10.000 334 0.029 0.002 30.000 4.651 335 0.385 0.085 21.221 4.795 10.000 336 0.524 0.119 7.682 20.000 5.115 10.000 337 0.266 30.000 14.328 5845 10 15 20 25 30 CA 03042004 2019-04-26 WO 2018/106284 PCT/US2017/033666 Ex. No. Flux Na/I.6 IC50 (pM) EP Nav1.6 IC50 (pM) Flux Na/L5 IC50(pM) EP Nav1.5 IC50 (pM) Flux Na/1.1 IC50 (pM) EP Nav1.1 IC50 (pM) 338 1.619 2.653 5.710 BIOLOGICAL EXAMPLE 3 Electrical Stimulation Seizure Assays Many electric stimulation seizure tests have been used to identify compounds with anti-convulsion activity, i.e., which raise seizure threshold. Two examples of electrical stimulation seizure assays frequently used in the field are the 6Hz psychomotor seizure assay (6Hz) and the Maximal Electroshock Seizure (MES). The 6Hz assay is considered a model of partial seizures observed in humans (Ldscher, W. and Schmidt, D., Epilepsy Res. (1988), Vol. 2, pp 145-81; Barton, M.E. etal., Epilepsy Res. (2001), Vol. 47, pp. 217-27). The MES assay is a model for generalized tonicclonic seizures in humans and provides an indication of a compound’s ability to prevent seizure spread when all neuronal circuits in the brain are maximally active. These seizures are highly reproducible and are electrophysiologically consistent with human seizures (Toman etal., 1946; Piredda etal., 1984; White etal., 1995). Experiments can be performed with healthy animals, or with seizure prone animals that have been genetically modified to model genetic epilepsy syndromes (Piredda, S. G. etal., J. Pharmacol. Exp. Ther. (1985), Vol. 232, pp. 741-5; Toman, J.E. etal., J. Neurophysiol. (1946), Vol. 9, pp. 231-9; and White, H.S. etal., Itai. J. Neurol. Sci. (1995), Vol. 16 (1- 2), pp. 73-7). To facilitate testing mice can be pretreated with the test compound or with the appropriate vehicle prior to the application of the electroshock. Each treatment group (n = 4-8 mice / group) is examined for anticonvulsive effects at different time points after administration of the compound and the vehicle. The eyes of mice are first anesthetized with a topical application of Alcaine (proparacaine hydrochloride) 0.5 %, one drop in each eye 30 min prior to the stimulation. Seizures are then induced by placing electrodes on the eyes which deliver a transcomeal current. The 6Hz Psychomotor Seizure test: Following pretreatment, each mouse is challenged with the low-frequency (6 Hz, 0.3 ms pulse width) stimulation for 3 sec. delivered through corneal electrodes at several intensities (12-44 mA). Animals are manually restrained and released immediately following the stimulation and observed for the presence or absence of 5855 10 15 20 CA 3042004 seizure activity. Typically, the 6 Hz stimulation results in a seizure characterized by a minimal clonic phase that is followed by stereotyped, automatist behaviors, including twitching of the vibrissae, and Straub-tail or by a generalized tonic clonic seizure. The presence, type and latency to seizure (in seconds) after the application of the current are monitored. Animals not displaying a clonic or generalized tonic clonic seizure are considered “protected”. All animals are euthanized at the end of assay. Plasma and brain samples are collected. Maximal Electroshock Test (MES): Following pretreatment, each mouse is challenged with an alternating current (60 Hz, 0.4-0.6 ms pulse width) for 0.2-0.5 sec. delivered through corneal electrodes at intensities (44- 55 mA). Typically, the MES stimulation results in a generalized tonic seizure that can be followed by a clonic seizure, automatist behaviors and Straub-tail. The presence, type and latency to seizure (in seconds) after the application of the current are monitored. An animal is considered “protected” from MES-induced seizures upon abolition of the hindlimb tonic extensor component of the seizure. After the seizure, mice are expected to resume normal exploratory behaviour within 1 to 4 minutes. Latency to seizure is recorded with a cut-off of 1 minute after which all animals are euthanized. ***** Although the foregoing invention has been described in some detail to facilitate understanding, it will be apparent that certain changes and modifications may be practiced within the scope of the appended claims. Accordingly, the described embodiments are to be considered as illustrative and not restrictive, and the invention is not to be limited to the details given herein, but may be modified within the scope and equivalents of the appended claims. 586 Date Regue/Date Received 2022-05-17DEMANDE OU BREVET VOLUMINEUX LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND PLUS D’UN TOME. CECI EST LE TOME 3 DE 3 CONTENANT LES PAGES 572 A 589 NOTE : Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE VOLUME THIS IS VOLUME 3 OF 3 CONTAINING PAGES 572 TO 589 NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME : NOTE POUR LE TOME / VOLUME NOTE:

Claims

CA 3042004 WHAT IS CLAIMED IS: 1. A compound, which is: 4-((2-(azetidin-1-ylmethyl)benzyl)amino)-5-chloro-2-fluoro-A/-(thiazol-4- yl)benzenesulfonamide; 4-((2-((2-oxa-6-azaspiro[3.3]heptan-6-yl)methyl)-6-fluorobenzyl)amino)-2,6- difluoro-A/-(thiazol-4-yl)benzenesulfonamide; 4-((2-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)-6-fluorobenzyl)amino)-2,6- difluoro-/V-(thiazol-4-yl)benzenesulfonamide; 4-((2-((tert-butyl(methyl)amino)methyl)-6-fluorobenzyl)amino)-2,6-difluoro-A/- (thiazol-4-yl)benzenesulfonamide; 4-((2-(azetidin-1-ylmethyl)benzyl)amino)-2,6-difluoro-A/-(thiazol-4- yl)benzenesulfonamide; (S)-4-((2-(1-(azetidin-1-yl)ethyl)-6-fluorobenzyl)amino)-2,6-difluoro-/V-(thiazol-4- yl)benzenesulfonamide; 4-((2-(azetidin-1-ylmethyl)-3,6-difluorobenzyl)amino)-2,6-difluoro-A/-(thiazol-4- yl)benzenesulfonamide; 2,3-difluoro-4-((2-fluoro-6-(pyrrolidin-1-ylmethyl)benzyl)amino)-/\/-(isoxazol-3- yl)benzenesulfonamide; 4-((2-((2,2-dimethylazetidin-1-yl)methyl)benzyl)amino)-2,6-difluoro-3-methyl-A/- (thiazol-4-yl)benzenesulfonamide; or 2,6-difluoro-4-((2-fluoro-6-((3-fluoro-3-methylazetidin-1-yl)methyl)benzyl)amino)- /V-(isoxazol-3-yl)benzenesulfonamide; or a pharmaceutically acceptable salt or solvate thereof.
2. The compound of claim 1, which is: 4-((2-(azetidin-1-ylmethyl)benzyl)amino)-5-chloro-2-fluoro-A/-(thiazol-4- yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof. 587 Date Re^ue/Date Received 2023-10-18CA 3042004
3. The compound of claim 1, which is: 4-((2-((2-oxa-6-azaspiro[3.3]heptan-6-yl)methyl)-6-fluorobenzyl)amino)-2,6- difluoro-A/-(thiazol-4-yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof.
4. The compound of claim 1, which is: 4-((2-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)-6-fluorobenzyl)amino)-2,6- difluoro-A/-(thiazol-4-yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof.
5. The compound of claim 1, which is: 4-((2-((tert-butyl(methyl)amino)methyl)-6-fluorobenzyl)amino)-2,6-difluoro-/V- (thiazol-4-yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof.
6. The compound of claim 1, which is: 4-((2-(azetidin-1-ylmethyl)benzyl)amino)-2,6-difluoro-/V-(thiazol-4- yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof.
7. The compound of claim 1, which is: (S)-4-((2-(1-(azetidin-1-yl)ethyl)-6-fluorobenzyl)amino)-2,6-difluoro-A/-(thiazol-4- yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof.
8. The compound of claim 1, which is: 4-((2-(azetidin-1-ylmethyl)-3,6-difluorobenzyl)amino)-2,6-difluoro-/V-(thiazol-4- yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof. 588 Date Re^ue/Date Received 2023-10-18CA 3042004
9. The compound of claim 1, which is: 2,3-difluoro-4-((2-fluoro-6-(pyrrolidin-1-ylmethyl)benzyl)amino)-/V-(isoxazol-3- yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof.
10. The compound of claim 1, which is: 4-((2-((2,2-dimethylazetidin-1-yl)methyl)benzyl)amino)-2,6-difluoro-3-methyl-A/- (thiazol-4-yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof.
11. The compound of claim 1, which is: 2,6-difluoro-4-((2-fluoro-6-((3-fluoro-3-methylazetidin-1-yl)methyl)benzyl)amino)- A/-(isoxazol-3-yl)benzenesulfonamide, or a pharmaceutically acceptable salt or solvate thereof.
12. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound of any one of claims 1-11, or a pharmaceutically acceptable salt or solvate thereof.
13. A compound of any one of claims 1-11, or a pharmaceutically acceptable salt or solvate thereof, for use in treatment of a disease or condition associated with Nad-6 activity in a mammal, wherein the disease or condition is epilepsy and/or epileptic seizure disorder.
14. A use of a compound of any one of claims 1-11, or a pharmaceutically acceptable salt or solvate thereof, for treatment of a disease or condition associated with Nad.6 activity in a mammal, wherein the disease or condition is epilepsy and/or epileptic seizure disorder.
15. A use of a compound of any one of claims 1-11, or a pharmaceutically acceptable salt or solvate thereof, for preparation of a medicament for treatment of a disease or condition associated with Nad.6 activity in a mammal, wherein the disease or condition is epilepsy and/or epileptic seizure disorder. 589 Date Re^ue/Date Received 2023-10-18CA 3042004
16. A compound, which is: 4-((2-(azetidin-1-ylmethyl)benzyl)amino)-5-chloro-2-fluoro-A/-(thiazol-4- yl)benzenesulfonamide; 4-((2-((2-oxa-6-azaspiro[3.3]heptan-6-yl)methyl)-6-fluorobenzyl)amino)-2,6- difluoro-A/-(thiazol-4-yl)benzenesulfonamide; 4-((2-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)-6-fluorobenzyl)amino)-2,6- difluoro-A/-(thiazol-4-yl)benzenesulfonamide; 4-((2-((tert-butyl(methyl)amino)methyl)-6-fluorobenzyl)amino)-2,6-difluoro-/V- (thiazol-4-yl)benzenesulfonamide; 4-((2-(azetidin-1-ylmethyl)benzyl)amino)-2,6-difluoro-A/-(thiazol-4- yl)benzenesulfonamide; (S)-4-((2-(1-(azetidin-1-yl)ethyl)-6-fluorobenzyl)amino)-2,6-difluoro-/V-(thiazol-4- yl)benzenesulfonamide; 4-((2-(azetidin-1-ylmethyl)-3,6-difluorobenzyl)amino)-2,6-difluoro-/V-(thiazol-4- yl)benzenesulfonamide; 2,3-difluoro-4-((2-fluoro-6-(pyrrolidin-1-ylmethyl)benzyl)amino)-N-(isoxazol-3- yl)benzenesulfonamide; 4-((2-((2,2-dimethylazetidin-1-yl)methyl)benzyl)amino)-2,6-difluoro-3-methyl-A/- (thiazol-4-yl)benzenesulfonamide; or 2,6-difluoro-4-((2-fluoro-6-((3-fluoro-3-methylazetidin-1-yl)methyl)benzyl)amino)- A/-(isoxazol-3-yl)benzenesulfonamide; or a pharmaceutically acceptable salt thereof.
17. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound of claim 16, or a pharmaceutically acceptable salt thereof.
18. A compound of claim 16, or a pharmaceutically acceptable salt thereof, for use in treatment of a disease or condition associated with Nay1.6 activity in a mammal, wherein the disease or condition is epilepsy and/or epileptic seizure disorder. 590 Date Re^ue/Date Received 2023-10-18CA 3042004
19. A use of a compound of claim 16, or a pharmaceutically acceptable salt thereof, for treatment of a disease or condition associated with Na/1.6 activity in a mammal, wherein the disease or condition is epilepsy and/or epileptic seizure disorder.
20. A use of a compound of claim 16, or a pharmaceutically acceptable salt thereof, for preparation of a medicament for treatment of a disease or condition associated with Na/1.6 activity in a mammal, wherein the disease or condition is epilepsy and/or epileptic seizure disorder.
21. A compound, which is selected from the group consisting of: 4-((2-(azetidin-1-ylmethyl)benzyl)amino)-5-chloro-2-fluoro-A/-(thiazol-4- yl)benzenesulfonamide; 4-((2-((2-oxa-6-azaspiro[3.3]heptan-6-yl)methyl)-6-fluorobenzyl)amino)-2,6- difluoro-A/-(thiazol-4-yl)benzenesulfonamide; 4-((2-((3-azabicyclo[3.1.0]hexan-3-yl)methyl)-6-fluorobenzyl)amino)-2,6- difluoro-A/-(thiazol-4-yl)benzenesulfonamide; 4-((2-((tert-butyl(methyl)amino)methyl)-6-fluorobenzyl)amino)-2,6-difluoro-/V- (thiazol-4-yl)benzenesulfonamide; 4-((2-(azetidin-1-ylmethyl)benzyl)amino)-2,6-difluoro-A/-(thiazol-4- yl)benzenesulfonamide; (S)-4-((2-(1-(azetidin-1-yl)ethyl)-6-fluorobenzyl)amino)-2,6-difluoro-/V-(thiazol-4- yl)benzenesulfonamide; 4-((2-(azetidin-1-ylmethyl)-3,6-difluorobenzyl)amino)-2,6-difluoro-/V-(thiazol-4- yl)benzenesulfonamide; 2,3-difluoro-4-((2-fluoro-6-(pyrrolidin-1-ylmethyl)benzyl)amino)-/V-(isoxazol-3- yl)benzenesulfonamide; 4-((2-((2,2-dimethylazetidin-1-yl)methyl)benzyl)amino)-2,6-difluoro-3-methyl-A/- (thiazol-4-yl)benzenesulfonamide; and 2,6-difluoro-4-((2-fluoro-6-((3-fluoro-3-methylazetidin-1-yl)methyl)benzyl)amino)- /V-(isoxazol-3-yl)benzenesulfonamide.
22. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound of claim 21. 591 Date Re^ue/Date Received 2023-10-18CA 3042004
23. A compound of claim 21 for use in treatment of a disease or condition associated with Na/I.6 activity in a mammal, wherein the disease or condition is epilepsy and/or epileptic seizure disorder.
24. A use of a compound of claim 21 for treatment of a disease or condition associated with Na/1.6 activity in a mammal, wherein the disease or condition is epilepsy and/or epileptic seizure disorder.
25. A use of a compound of claim 21 for preparation of a medicament for treatment of a disease or condition associated with Na/1.6 activity in a mammal, wherein the disease or condition is epilepsy and/or epileptic seizure disorder. 592 Date Re^ue/Date Received 2023-10-18