, Using the standard procedure from 1-phenyl-2-(1H-pyrazol-1-yl)ethan-1-ol 4a (0.188 g, 1 mmol) and 1-(3-bromopropyl)-4-methoxybenzene 5c (0.229 g, 1 mmol, 1 equiv.), the 1-[?-((3-phenylpropyl)oxy)-phenethyl]-1H-pyrazole 6b (0.151 g, 0.45 mmol) was dissolved in 3 mL of dry diethyl ether pa under a stream of argon. The addition of 0.45 mL of 1M HCl (0.45 mmol, 1 equiv, propyl)oxy)-phenethyl]-1H-pyrazolium hydrochloride (7b), p.72

?. , -methoxyphenyl)propyl)oxy)-phenethyl]-1H-pyrazolium hydrochloride 7b (78.3 mg, 21% yield) was obtained as white powder

1. Hz-;-dt, J. Hz, C. H. , H. Dt, 1. et al., 113.6 (C-3b, C-5b), 126.7 (C-2a, C-6a), 128.0 (C-4a), vol.2, 1916.

, g, 1 mmol) and 4-methoxybenzyl chloride 5a (0.157 g, 1 mmol, 1 equiv.), the 1-[?-((4-methoxybenzyl)oxy)-(4-methoxyphenethyl]-1H-pyrazole 6c (0.231 g, 0.68 mmol) , the 1-[?-((4-methoxybenzyl)oxy)-(4-methoxyphenethyl]-1H-pyrazolium hydrochloride 7c (123.7 mg, 33% yield) was obtained as white powder. Mp = 112-113 ? C. 1 H NMR (DMSO-d 6 ) ? = 3.72 (s, 3H, 1H-pyrazolium hydrochloride (7c). Using the standard procedure from 1-(4-methoxyphenyl

H. H-6a and H. , Ar), 7.62 (d, 1H, J = 2.2 Hz, H-5, Ar). 13 C NMR (DMSO-d 6 ) ? = 55, 158.6 (C-4a), 159.1 (C-4b). ES + HRMS, m/z = 361.1526 found, vol.55

, Using the standard procedure from 1-(4-methoxyphenyl)-2-(1H-pyrazol-1-yl)ethan-1-ol 4b (0.218 g, 1 mmol) and 1-(3-bromopropyl)-4-methoxybenzene 5c (0.229 g, 1 mmol, 1 equiv.), the 1

, ) produced a pale pink viscous oil that crystallized (12 h) on the circumference of the round flask, and mixing (200 rpm, 72 h) was pursued until complete crystallization of the viscous gum. The desired salt 7d was collected by filtration on a, Büchner Int. J. Mol. Sci, vol.19, pp.856-873, 2018.

, Ar). 13 C NMR (DMSO-d 6 ) ? = 30.4 (C-3"), 31.1 (C-2"), mg, 18% yield) was obtained as white powder. Mp = 92-94 ? C. 1 H NMR (DMSO-d 6 ) ? = 1.47-1.70 (m, 2H, CH 2 , H-2"), 2.36 (t, 2H, J = 7.5 Hz, vol.2

, 1R, 1S) 1-[?

, Using the standard procedure from 1-(4-methoxyphenyl)-2-(1H-pyrazol-1-yl)ethan-1-ol 4b (0.218 g, 1 mmol) and 1-bromo-3-phenylpropane 5d (0.199 g, 1 mmol, 1 equiv.), the 1

. ?-c-;-m,-2h, C. H. Hz, and H. , Ar). 13 C NMR (DMSO-d 6 ) ? = 30, mg, 22% yield) was obtained as white powder. Mp = 87-89 ? C. 1 H NMR (DMSO-d 6 ) ? = 1.54-1.76 (m, 2H, CH, vol.2

, -(1H-pyrazol-1-yl)ethan-1-ol (4b) with (?)-(1R)-10-Camphorsulfonic Acid and (+)-(1S)-10-Camphorsulfonic Acid by Two Convergent Methods of Half-Quantities Method 1 from (?)-(1R)-CSA: (?)-(1S) 1-(4-methoxyphenyl, Resolution of (±)-(1R, 1S) 1-(4-Methoxyphenyl

, This salt was recrystallized in dry acetone and afforded 0.271 g (52% isolated yield) of salt (?)-(1S)-4b/(?)-(1R)-CSA (>97% de). This salt was dissolved in 0.6 mL of dry acetone under magnetic stirring (300 rpm

, This salt was dissolved in 0.6 mL of dry acetone under magnetic stirring (300 rpm) and to this suspension was added 0.02 equiv. of racemic (±) 1-(4-methoxyphenyl)-2-(1H-pyrazol-1-yl)ethan-1-ol 4b for diastereomeric enrichment. This suspension was stirred for 18 h at room temperature. The enriched salt was successively filtered on a Büchner funnel, washed with 0.5 mL of dry acetone, 54% isolated yield) of salt (+)-(1R)-4b/(+)-(1S)-CSA (>96% de)

=. +6, , p.0

, Starting from 200 mg of the pure salt (+)-(1R)-4b/(+)-(1S)-CSA, a mixed suspension was

, × 2 mL of deionized water. The insoluble compound was collected by filtration on a Büchner funnel (porosity N ? 4) and dried in vacuum, which gave 0.089 g (35% yield) of the desired (+)-(1R) 1-(4-methoxyphenyl

, CSA (0.219 g, 0.95 mmol, 1 equiv.) in 1.66 mL of dry acetone was added dropwise. During the addition of the (?)-(1R)-CSA solution, a fine suspension appeared in the mixture, which was stirred at room temperature for 12 h. The new diastereomer salt was collected by filtration then washed with 4 × 0.25 mL of dry acetone and dried in vacuum, which gave 0.285 g (66% yield) of the (?)-(1S)-4b/(?)-(1R)-CSA salt (98% de) as white powder. From this salt, diastereomeric enrichment was conducted with 0.01 equiv. of racemic (±) 1-(4-methoxyphenyl)-2-(1H-pyrazol-of dry acetone, drying in vacuum, and finally recrystallisation in dry acetone which gave 0.258 mg (60% yield) of (?)-(1S), Recovery of the (?)-(1S) 1-(4-methoxyphenyl)-2-(1H-pyrazol-1-yl)ethan-1-ol 4b: The filtrate of the first to the resulting solution, a solution of (?)-(1R)

, Starting from 200 mg of the pure salt (?)-(1S)-4b/(?)-(1R)-CSA, a mixed suspension was

, × 2 mL of deionized water. The insoluble compound was collected by filtration on a Büchner funnel (porosity N ? 4) and dried in vacuum which gave 0.090 g (43% yield) of the desired (?)-(1S) 1-(4-methoxyphenyl

C. 3h, C. H. --;-m,-1h, H. --;-m,-1h, C. H. , and H. , 04 (s, 3H, C-7"CH 3 ), 1.19-1.40 (m, 2H, CH 2 , H-2, -(1S)-4b/(?)-(1R)-CSA salt: Yield = 48%. White powder, Mp = 168-170 ? C. [? D ] = ?6.0 (c 1.0, MeOH) and >99% de (retention time t R = 34.01 min. using hexane/i-PrOH 94:6 v/v as eluent, flow rate = 0.8 mL/min.). 1 H NMR (DMSO-d 6 ) ? = 0.75 (s, vol.1, p.24, 2018.

. Ar, 158.5 (C-4a), 216.0 (C=O). ES + HRMS, m/z = 241.0946 found, vol.47

, White powder, Mp = 169-171 ? C, -(1R)-4b/(+)-(1S)-CSA salt: Yield = 51%

3. , C. --;-s,-3h, C. --;-m,-2h, 1. , J. Hz et al.,

, X-ray crystallographic data for (+)-(1R)-4b/(+)-(1S)-CSA salt

, M = 450.54. APEXII, Bruker-AXS diffractometer, Mo-K? radiation (? = 0.71073 Å), T = 150

Å. and V. , ) Å 3 . Z = 4, d = 1.314 g·cm ?3 , µ = 0.182 mm ?1 . The structure was solved by direct methods using the SIR97 program [29], and then refined with full-matrix least-square methods based on F2 (SHELXL-97) [30] with the aid of the WINGX [31] program. All non-hydrogen atoms were refined with anisotropic atomic displacement parameters. Except oxygen-and nitrogen-linked hydrogen atoms that were introduced in the structural model through Fourier difference maps analysis, H atoms were finally included in their calculated positions. A final refinement on F 2 with 4854 unique intensities and 289 parameters converged at ?R(F 2 ) = 0.0932 (R(F) = 0.0415) for 4347 observed reflections with I > 2?(I), #19), a = 7.2781(5), b = 9, vol.21, pp.21-21

, MeOH) and >99% de (retention time t R = 52.6 min. using hexane/i-PrOH 96:4 v/v as eluent, flow rate = 0.6 mL/min.). 1 H NMR (DMSO-d 6 ) ? = 3.73 (s, 3H, -(1S) 1-(4-Methoxyphenyl)-2-(1H-pyrazol-1-yl)ethan-1-ol (4b): Yield = 38%. White powder, vol.6

, C-4 ), 113.5 (C-3a, C-5a), 127.2 (C-2a, C-6a), 130.5 C-5 ), 134.8 (C-1a), 158.5 (C-4a). ES + HRMS, m/z = 241.0950 found, vol.104

, MeOH) and >99% de (retention time t R = 63.5 min. using hexane/i-PrOH 96:4 v/v as eluent, flow rate = 0.6 mL/min.). 1 H NMR (DMSO-d 6 ) ? = 3.73 (s, 3H, -(1R) 1-(4-Methoxyphenyl)-2-(1H-pyrazol-1-yl)ethan-1-ol (4b): Yield = 41%. White powder, vol.19, pp.856-877, 2018.

, 158.5 (C-4a). ES + HRMS, m/z = 241.0950 found, vol.138

D. Sulfoxyde, Ref: 354241). Fura-2 QBT? Calcium Kit was purchased from Molecular Devices, DMSO-Ref D2438) and Thapsigargin (Tg-Ref: T9033) was purchased from Sigma-Aldrich. Corning ® Cell-Tak? was purchased from Beckton Dickinson

, The B-cell leukemia cell line PLP [32], which possesses the phenotypic characteristics of B-CLL cells [33], were maintained in RPMI-1640 containing 10% fetal calf serum and antibiotics (1% of penicillin/streptomycin)

, Cytosolic Ca 2+ Protocol For Ca 2+ experiments, cells were seeded overnight into four wells plate (Nunc TM multidisches 4 wells flat-bottom-Ref: 176740) at 7 × 10 5 cells/mL in 1 mL culture medium. Cell suspension was then transfered into 1.5 mL Microtube, pelleted, and loaded with Fura-2 acetoxymethyl ester (Fura-2 QBT?) fluorochrome according to the manufacturer's protocol. Cell suspension was dispensed onto Corning ® Cell-Tak? pre-coated 96 wells black clear bottom plate (80 µL per wells, density approximatively 5 × 10 4 cells per well) and incubated 1 h at 37 ? C, 5% CO 2 . The Fura-2 QBT? was aspirated and replaced by an equal volume of free Ca, p.135

. Kcl, UK), a fluorescence plate reader measuring time-resolved intracellular Ca 2+ concentration in a 96-well format with the dual-wavelength fluorescent calcium-sensitive dye Fura-2AM. Dual excitation wavelength capability permits ratiometric measurements of Fura-2AM peak emissions (510 nm) after excitations at 340 (bound to calcium) and 380 nm (unbound to Ca 2+ ). Modifications in the 340/380 ratio reflect changes in intracellular-free Ca 2+ concentrations. The FlexStation 3? temperature was setting at 37 ? C during data acquisition. Thapsigargin (2 µM) solution and Hepes-buffered solution with Ca 2+ containing (in mM): 135 NaCl, 5 KCl, 1 MgCl 2 , 2 CaCl 2 , 10 Hepes, 10 glucose, pH adjusted at 7.45 with NaOH were added from a 96-well reservoir plate during Calcium Mobilization Assay running at 100 and 750 s, respectively. Experimental setup parameters were optimized (pipette heights, volumes and rate of additions) to minimize disturbance of the cells while ensuring rapid mixing. The data were stored for later analysis by using SoftmaxPro, The tested compounds of Intracellular Calcium Levels Intracellular calcium levels were monitored by using the FlexStation 3? (Molecular Devices

, Measurement of Store-Operated Calcium (SOC) Entry

, Intracellular Ca 2+ stores were depleted with 2 µM Thapsigargin an inhibitor of ER SERCA pumps under Ca 2+ -free conditions to determine the magnitude of intracellular Ca 2+ release. Next, cells were returned to Ca 2+ -containing Hepes-buffered solution to measure SOCE. The magnitude of SOCE was estimated as the maximal values of normalized F340/F340 ratio following Ca 2+ re-addition

, The number of experiments for each group is represented as n. Data analysis for functional assays was performed using Softmax Pro and Graph Pad Prism. (Graph Pad): Ca 2+ concentration variations are estimated using the ratio of peak RFU at 340 and 380 nm (F340/F380) and for each measurement F340/F380 ratio values are normalized to the initial basal ratio before TG addition. Results are expressed in terms of percentage of response or percentage of inhibition compared to the maximum response on control condition. Inhibition is estimated as follow: % Inhibition = (100 ? % response). The EC 50 values were estimated from the dose/response curves (log(inhibitor) vs. amplitude of the Ca 2+ response) using a regression model, Int. J. Mol. Sci, vol.19, 2018.

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