N-Benzoyl-L-alaninol

FT-Raman and FT-IR spectra of N-{(meta-ferrocenyl) Benzoyl} - l-alanine - glycine ethyl ester were recorded in solid phase. The optimized molecular geometry, the vibrational wavenumbers, the infrared intensities and the Raman scattering intensities were calculated by using density functional method(B3LYP) with 6-31G(d, p) basis set. Vibrational assignment of the molecule was done by using potential energy distribution analysis. Natural bond orbital analysis, Mulliken charge analysis and HOMO-LUMO energy were used to elucidate the reasons for intra molecular charge transfer. Docking studies were conducted to predict its anticancer activity.

Beta-benzoyl-DL-alanine was synthesized from alpha-bromoacetophenone and diethyl acetamidomalonate. The racemic amino acid was resolved by carboxypeptidase A-catalyzed hydrolysis of the N-trifluoroacetyl derivative. Beta-benzoyl-L-alanine is a good substrate of kynureninase from Pseudomonas fluorescens, with k(cat) and k(cat)/K(m) values of 0.7 s(-1) and 8.0 x 10(4) M(-1) s(-1), respectively, compared to k(cat) = 16.0 s(-1) and k(cat)/K(m) = 6.0 x 10(5) M(-1) s(-1) for L-kynurenine. In contrast to the reaction of L-kynurenine, beta-benzoyl-L-alanine does not exhibit a significant solvent isotope effect on k(cat) ((H)k/(D)k = 0.96 +/- 0.06). The pre-steady-state kinetics of the reaction of beta-benzoyl-L-alanine were investigated by rapid scanning stopped-flow spectrophotometry. The spectra show the formation of a quinonoid intermediate, with lambda(max) = 490 nm, in the dead time of the instrument, which then decays, with k = 210 s(-1), to form a transient intermediate with lambda(max) at 348 nm. In the presence of benzaldehyde, the 348 nm intermediate decays, with k = 0.7 s(-1), to form a quasistable quinonoid species with lambda(max) = 492 nm. Previous studies demonstrated that benzaldehyde can trap an enamine intermediate formed after the C(beta)-C(gamma) bond cleavage [Phillips, R. S., Sundararaju, B., and Koushik, S. V. (1998) Biochemistry 37, 8783-8789]. Thus, the 348 nm intermediate is kinetically competent. The position of the absorption maximum and shape of the band is consistent with a PMP-ketimine intermediate. The results from chemical quenching analysis do not show a burst of benzoate and, thus, also support the formation of benzoate as the rate-determining step. These data suggest that, in contrast to L-kynurenine, for which the rate-determining step was shown to be deprotonation of the pyruvate-ketimine intermediate [Koushik, S. V., Moore, J. A., III, Sundararaju, B., and Phillips, R. S. (1998) Biochemistry 37, 1376-1382], the rate-determining step in the reaction of beta-benzoyl-L-alanine with kynureninase is C(beta)-C(gamma) bond cleavage.

N-Benzoyl-L-tyrosyl-1-13C-L-alanine sodium is a dipeptide used for evaluating pancreatic exocrine function. The method of evaluation, however, is not yet satisfactory, especially in experimental animals. The relation between diabetes and pancreatic exocrine function also is not clear. Therefore, this study sought to establish a method for evaluating pancreatic exocrine function and to validate the method by determining non-invasively the effect of alloxan-induced diabetes in conscious rats. After fasting, rats were orally administered Racol containing N-benzoyl-L-tyrosyl-1-13C-L-alanine sodium or 1-13C-L-alanine and housed in desiccators. The expired air in the desiccator was collected in a breath-sampling bag using a tube and aspiration pump, and the level of 13CO2 in this air was measured using an infrared spectrometer at appropriate intervals over a 120 min period. The rate of 13CO2 excretion increased, peaked and then decreased in a dose-dependent manner. The maximum concentration and area under the curve of 13CO2 excretion significantly and positively correlated with the doses of N-benzoyl-L-tyrosyl-1-13C-L-alanine sodium. In the rats made diabetes by the administration of alloxan, the level of expired 13CO2 air changed at significantly lower levels as compared with that of the control rats on day 3, although the level of expired 13CO2 air from 1-13C-L-alanine was also markedly lower than that of the control rats. These results showed that pancreatic exocrine function can be evaluated using this breath test system and that alloxan-induced diabetes affects this function.