International Congress Series 1245 (2002) 355 – 356

Kinetics and mechanism of the reaction of

metformin with methylglyoxal
Sinan Battah, Naila Ahmed, Paul J. Thornalley *
Department of Biological and Chemical Sciences, University of Essex, Central Campus,
Wivenhoe Park, Colchester, Essex CO4 3SQ, UK

Abstract

We examined the rate of reaction of methylglyoxal with metformin, forming hydroimidazolone,

triazepinone and other adducts at pH 7.4 and 37 jC in an initial rate study. Metformin was not an
efficient scavenger of methylglyoxal. Mechanisms other than the scavenging of methylglyoxal
were probably responsible for decreased methylglyoxal concentration in diabetic subjects on
metformin therapy—the lifting of insulin resistance, for example.
D 2002 Elsevier Science B.V. All rights reserved.

Keywords: Metformin; Methylglyoxal; Glycation; Hydroimidazolone; Triazepinone

1. Introduction

Metformin is a drug used in the therapy of type 2 diabetes mellitus. It decreased blood
glucose levels by increasing hepatic and peripheral tissue sensitivity to insulin [1]. It was
also found recently to decrease the blood plasma concentration of methylglyoxal [2].
Metformin has been proposed as a scavenger of methylglyoxal in vivo where a
triazepinone adduct was formed [3].

2. Results and discussion

The reaction of methylglyoxal and metformin was associated with an initial rapid
increase in absorbance at 225 nm and formation of early stage adducts. Purification of

Abbreviations: MALDI, matrix-assisted laser desorption ionisation.

*
Corresponding author. Tel./fax: +44-1206-873010.
E-mail address: thorp@essex.ac.uk (P.J. Thornalley).

0531-5131/02 D 2002 Elsevier Science B.V. All rights reserved.

PII: S 0 5 3 1 - 5 1 3 1 ( 0 2 ) 0 0 8 8 9 - 0
356 S. Battah et al. / International Congress Series 1245 (2002) 355–356

Fig. 1. Adducts formed by the reaction of methylglyoxal with metformin.

these adducts by reversed phase HPLC gave a 1H NMR spectrum and matrix-assisted
laser desorption ionisation (MALDI) mass spectrometric data indicative of a mixture of
hydroimidazolone structural isomers and tautomers, and a minor imidazolidine compo-
nent (Fig. 1). The mean e255 was 13,343 F 24 M 1 cm 1. The rate of formation of the
hydroimidazolones was studied with 3 –300 AM metformin and 5 – 500 AM methyl-
glyoxal. The initial rate r0 = k[methylglyoxal][metformin], where the rate constant
k = 0.56 F 0.14 M 1 min 1. The formation of the triazepinone was monitored by
absorbance at 335 nm (e335 = 38.2 F 0.2 M 1 cm 1). The slower rate of formation of
the triazepinone was studied with 1– 25 mM metformin and 2 –20 mM methylglyoxal.
The reaction was first order with respect to methylglyoxal; it was also first order with
respect to metformin at low metformin concentrations but zeroth order at high metformin
concentrations (where the rate of methylglyoxal dehydration was rate limiting). Kinetic
analysis gave k values: for the reaction of unhydrated methylglyoxal CH3COCHO with
metformin, k = 3.28 F 0.15 M 1 min 1; for the reaction of methylglyoxal monohydrate
CH3COCH(OH)2 with metformin, k=(1.4 F 0.1)  10 4 M 1 min 1; for the reaction of
methylglyoxal (all forms) with metformin, k = 0.034 F 0.002 M 1 min 1. Aminogua-
nidine was a much better scavenger of methylglyoxal: k for methylglyoxal (all forms) was
155 M 1 min 1—ca. 450 fold higher than for metformin.

References

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glyoxal levels in type 2 diabetes, Diabetes 48 (1999) 198 – 202.
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