Diabetes Metab J.  2020 Dec;44(6):908-918. 10.4093/dmj.2019.0147.

Role of Intestinal Microbiota in Metabolism of Voglibose In Vitro and In Vivo

• Nepal MR
• Kang MJ
• Kim GH
• Cha DH
• Kim JH
• Jeong TC

• KMID: 2513054
• DOI: http://doi.org/10.4093/dmj.2019.0147

Abstract

Voglibose, an α-glucosidase inhibitor, inhibits breakdown of complex carbohydrates into simple sugar units in intestine. Studies showed that voglibose metabolism in the liver might be negligible due to its poor intestinal absorption. Numerous microorganisms live in intestine and have several roles in metabolism and detoxification of various xenobiotics. Due to the limited information, the possible metabolism of voglibose by intestinal microbiota was investigated in vitro and in vivo.

For the in vitro study, different concentrations of voglibose were incubated with intestinal contents, prepared from both vehicle- and antibiotics-treated mice, to determine the decreased amount of voglibose over time by using liquid chromatography-mass spectrometry. Similarly, in vivo pharmacodynamic effect of voglibose was determined following the administration of voglibose and starch in vehicle- and antibiotic-pretreated non-diabetic and diabetic mice, by measuring the modulatory effects of voglibose on blood glucose levels.

The in vitro results indicated that the remaining voglibose could be significantly decreased when incubated with the intestinal contents from normal mice compared to those from antibiotic-treated mice, which had less enzyme activities. The in vivo results showed that the antibiotic pretreatment resulted in reduced metabolism of voglibose. This significantly lowered blood glucose levels in antibiotic-pretreated mice compared to the control animals.

The present results indicate that voglibose would be metabolized by the intestinal microbiota, and that this metabolism might be pharmacodynamically critical in lowering blood glucose levels in mice.

Keyword

Anti-bacterial agents; Blood glucose; Gastrointestinal microbiome; Metabolism; Voglibose

Figure

• 

  Fig. 1 Mass spectra of (A) voglibose with its structure and (B) telmisartan, an internal standard. cps, counts per second; m/z, mass to charge.

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  Fig. 2 Representative liquid chromatography-mass spectrometry (LC-MS/MS) chromatograms of (A, B, C) voglibose and (D, E, F) telmisartan. (A, D) blank samples; (B, E) blank samples spiked with analyte at lower limit of quatitation; and (C, F) real samples from in vitro incubation with intestinal contents for 3 hours. cps, counts per second.

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  Fig. 3 Enzyme activities of intestinal contents: 0.4 mL of either 2.5 mM 4-nitrophenyl β-D-glucopyranoside, 4-nitrophenyl β-D-glucuronide, or 4-nitrophenyl-sulfate in potassium phosphate buffer, pH 7.4, was mixed with 0.4 mL of potassium phosphate buffer, and 0.2 mL of intestinal contents prepared from both antibiotics- and vehicle-treated mice and incubated at 37℃ for 30 minutes. The enzyme activities were calculated following measuring the sample's absorbance at 405 nm. Each bar represents the mean activity of respective enzymes+standard deviation of triplicate determinations. aThe value significantly different from corresponding vehicle-treated controls at P<0.05.

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  Fig. 4 Time- and concentration-dependent metabolism of voglibose by intestinal microbiota in vitro. Various concentrations of voglibose was incubated with 0.5 g/mL intestinal contents prepared from both antibiotics- and vehicle-treated mice and incubated at 37℃ for various time points. The remained voglibose over time was measured by liquid chromatography-mass spectrometry (LC-MS/MS). Each bar represents the mean concentration of voglibose in the incubated sample+standard deviation of triplicate determinations. aThe value significantly different from corresponding vehicle-treated controls at P<0.05.

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  Fig. 5 Blood glucose level in vehicle-treated and antibiotics-treated non-diabetic normal mice (A) and diabetic mice (B). Six-hr fasted vehicle- and antibiotics-treated non-diabetic and diabetic mice were administered with voglibose followed by an oral administration with starch. And then, the change in the blood glucose level was measured at various time points. Each point represents mean blood glucose level±standard deviation of five animals. aThe value indicates significant difference between antibiotics-treated starch only and antibiotics-treated starch+voglibose groups, bThe value indicates significant difference between vehicle-treated starch+voglibose and antibiotics-treated starch+voglibose groups.

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