Enantioseparation of basic pharmaceutical compounds by capillary electrophoresis using sulfated cyclodextrins. Application to E-6006, a novel antidepressant.
In this study, a chiral capillary electrophoresis method was optimized and validated for E-6006, a thienylpyrazolylethanamine derivative (pKa 8.9). Enantioselectivity of neutral and anionic cyclodextrins (CDs) was evaluated at acid pH (3), obtaining cathodic and anodic migration, respectively. Hydroxypropyl-beta-CD, carboxymethyl-beta-CD and sulfobutyl ether-beta-CD led to similar and partial selectivity, whereas sulfate (S)-beta-CD produced baseline separation of the enantiomers. Four types of sulfated CDs were compared considering: cavity size (alpha, beta, gamma) and random substitution versus unique derivative (S-beta-CD, 6-heptakis-S-beta-CD). Complete peak separation was obtained in all cases, but with different affinity and binding strength. Some factors that play a role in the complex formation include: position/region/degree of substitution, size of CD cavity and proportion of derivatives in mixtures. Enantioaffinity and enantioselectivity increased with the average of sulfate groups/mol. Beta cavity size complexed better, although alpha and gamma cavities did not compromise separation. 6-Heptakis-S-beta-CD had less affinity and separation efficiency, attributed to its lower degree and unique position of substitution. The method was optimized with S-beta-CD (Aldrich, randomly substituted, 7-11 groups/mol). With this selector, the effect of pH value (3-9) was evaluated. Around pH 7 the cross-over point with change in the direction and order of migration was observed, associated with great enantioselectivity and long migration times. Fine tuning was done by adjusting the CD concentration and the buffer counterion. Definitive conditions were: uncoated silica capillary, 10 mM S-beta-CD-25 mM sodium phosphate, pH 3. Validation parameters are included.
Gómez-Gomar
A
A
Medicinal Chemistry Department, Laboratorios Dr. Esteve S.A., Av. V. Montserrat 221, 08041 Barcelona, Spain. agomez@esteve.es
Ortega
E
E
Calvet
C
C
Andaluz
B
B
Mercé
R
R
Frigola
J
J
D000928:Antidepressive Agents / Q000302:isolation & purification*; D002138:Calibration; D003505:Cyclodextrins / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D011720:Pyrazoles / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism
C427705:5-(alpha-(2-(dimethylamino)ethoxy)-2-thienylmethyl)-1-methyl-1H-pyrazole; D000928:Antidepressive Agents; D003505:Cyclodextrins; D011720:Pyrazoles