Preparation of MP compounds
Compounds MP-001 and MP-002, strategically designed to target the interaction site of FKBP12/RyR1, were synthesized using CuAAC ”click chemistry” methodologies. Specifically, aryl propargyl sulfides and methyl azidoglycinate were utilized for MP-001, while 2-azidoethyl-N,N-dimethylammonium hydrochloride was employed for MP-002 synthesis . Compound MP-010 was prepared by adding phthaloyl peroxide to a solution of MP-002, followed by stirring at room temperature, basification with 7 N NH3 in MeOH, and evaporation under reduced pressure. The resulting crude product underwent purification through column chromatography (silica gel; eluent: CH2Cl2 / MeOH (7N NH3)). The yield of the purified white solid was 66%, with1H and 13C NMR spectra confirming the chemical structure: 1H NMR (400 MHz, CD3OD) δ 7.81 (s, 1H), 7.49 (d, J = 8.8 Hz, 2H), 7.95 (d, J = 8.8 Hz, 2H), 4.51 (t, J = 6.3 Hz, 2H), 4.30 (dd, J = 13.7, 1.6 Hz, 2H), 3.86 (s, 3H), 2.82 (t, J = 6.3 Hz, 2H), 2.31 (s, 6H). 13C NMR (101 MHz, CD3OD) δ 164.1, 137.5, 133.5, 127.7, 126.7, 115.9, 59.5, 56.1, 53.6, 45.4. IR (cm-1): 1592, 1495, 1459, 1303, 1249, 1172, 1086, 1022, 829, 524.
To obtain aqueous 10 mM stock solutions of these compounds for pharmaceutical application, the free carboxylic acid of MP-001 or the tertiary amine moiety of MP-010 was neutralized to pH = 6.8 using 2 M NaOH and 2 M HCl, respectively. These neutralized solutions were further diluted with drinking water to achieve pharmaceutically acceptable concentrations of 1 mM or 2 mM prior to administration.