The Glitazone Class of Drugs as Carbonic Anhydrase Inhibitors-A Spin-Off Discovery from Fragment Screening.

The approved drugs that target carbonic anhydrases (CA, EC 4.2.1.1), a family of zinc metalloenzymes, comprise almost exclusively of primary sulfonamides (R-SO2NH2) as the zinc binding chemotype. New clinical applications for CA inhibitors, particularly for hard-to-treat cancers, has driven a growing interest in the development of novel CA inhibitors. We recently discovered that the thiazolidinedione heterocycle, where the ring nitrogen carries no substituent, is a new zinc binding group and an alternate CA inhibitor chemotype. This heterocycle is curiously also a substructure of the glitazone class of drugs used in the treatment options for type 2 diabetes. Herein, we investigate and characterise three glitazone drugs (troglitazone 11, rosiglitazone 12 and pioglitazone 13) for binding to CA using native mass spectrometry, protein X-ray crystallography and hydrogen-deuterium exchange (HDX) mass spectrometry, followed by CA enzyme inhibition studies. The glitazone drugs all displayed appreciable binding to and inhibition of CA isozymes. Given that thiazolidinediones are not credited as a zinc binding group nor known as CA inhibitors, our findings indicate that CA may be an off-target of these compounds when used clinically. Furthermore, thiazolidinediones may represent a new opportunity for the development of novel CA inhibitors as future drugs.

Detection of structurally similar adulterants in botanical dietary supplements by thin-layer chromatography and surface enhanced Raman spectroscopy combined with two-dimensional correlation spectroscopy.

Thin-layer chromatography (TLC) coupled with surface enhanced Raman spectroscopy (SERS) has been widely used for the study of various complex systems, especially for the detection of adulterants in botanical dietary supplements (BDS). However, this method is not sufficient to distinguish structurally similar adulterants in BDS since the analogs have highly similar chromatographic and/or spectroscopic behaviors. Taking into account the fact that higher cost and more time will be required for comprehensive chromatographic separation, more efforts with respect to spectroscopy are now focused on analyzing the overlapped SERS peaks. In this paper, the combination of a TLC-SERS method with two-dimensional correlation spectroscopy (2DCOS), with duration of exposure to laser as the perturbation, is applied to solve this problem. Besides the usual advantages of the TLC-SERS method, such as its simplicity, rapidness, and sensitivity, more advantages are presented here, such as enhanced selectivity and good reproducibility, which are obtained by 2DCOS. Two chemicals with similar structures are successfully differentiated from the complex BDS matrices. The study provides a more accurate qualitative screening method for detection of BDS with adulterants, and offers a new universal approach for the analysis of highly overlapped SERS peaks.

Rapid on-site TLC-SERS detection of four antidiabetes drugs used as adulterants in botanical dietary supplements.

A novel facile method has been established for rapid on-site detection of antidiabetes chemicals used to adulterate botanical dietary supplements (BDS) for diabetes. Analytes and components of pharmaceutical matrices were separated by thin-layer chromatography (TLC) then surface-enhanced Raman spectroscopy (SERS) was used for qualitative identification of trace substances on the HPTLC plate. Optimization and standardization of the experimental conditions, for example the method used for preparation of silver colloids, the mobile phase, and the concentration of colloidal silver, resulted in a very robust and highly sensitive method which enabled successful detection when the amount of adulteration was as low as 0.001 % (w/w). The method was also highly selective, enabling successful identification of some chemicals in extremely complex herbal matrices. The established TLC-SERS method was used for analysis of real BDS used to treat diabetes, and the results obtained were verified by liquid chromatography-triple quadrupole mass spectrometry (LC-MS-MS). The study showed that TLC-SERS could be used for effective separation and detection of four chemicals used to adulterate BDS, and would have good prospects for on-site qualitative screening of BDS for adulterants.

[Detection of glimepiride and rosiglitazone from Chinese dietary supplements].

A 79-year-old man with diabetes mellitus developed prolonged hypoglycemia. The patient had ingested two Chinese dietary supplements in addition to his prescribed nateglinide (Fastic). Using liquid chromatography tandem mass spectrometry, glimepiride from sulfonylurea, as well as rosiglitazone from a thiazolidine derivative, were detected in the Chinese dietary supplements, which were then quantitatively analyzed using liquid chromatography with UV detector. Mean values (n=3) of glimepiride contents of the Chinese dietary supplements were 0.75 and 0.86 mg/capsule. Predicted intake of glimepiride in the patient was estimated to be 4.8-8.2 mg/day according to the glimepiride contents and directions of the Chinese dietary supplements. The daily intake of glimepiride in this patient was greater than daily maintenance doses (1-4 mg) of glimepiride for diabetic patients. Therefore, overdose of glimepiride by ingestion of the Chinese dietary supplements appears to be associated with the development of prolonged hypoglycemia.

Evaluation of solid state form of troglitazone by solid state NMR spectroscopy.

The solid state forms of troglitazone drug substance and diastereomers were characterized using solid state nuclear magnetic resonance (SSNMR) spectroscopic method. The SSNMR spectroscopy could distinguish the hydrated and the non-hydrated RR/SS forms more clearly than powder X-ray diffractometry (PXRD). The SSNMR result supported that troglitazone drug substance consists of diastereomers as a simple physical mixture. SSNMR spectroscopy was also able to characterize the solid state forms of troglitazone in tablets while PXRD was unable to because of interference from the pharmaceutical additives. Troglitazone was proved to exist in amorphous form in tablets, and keep its solid state form amorphous against heat and humidity. SSNMR spectroscopy thus provides very important information for the development of the pharmaceutical formulation of troglitazone.