Laboratory of Bioanalysis, Department of Pharmaceutical Chemistry, Drug Analysis and Radiopharmacy, Medical University of Lodz, ul. Muszyńskiego1, 90-151 Lodz, Poland.
School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Yliopistonranta 1C, POB 1627, 70211 Kuopio, Finland.
Chem Biol Interact. 2018 Mar 25;284:126-136. doi: 10.1016/j.cbi.2018.02.012. Epub 2018 Feb 16.
Type 2 diabetes mellitus (T2DM) is characterised not only by hyperglycaemia and insulin resistance but also an impaired balance between the processes of coagulation and fibrinolysis. The aim of this study was to examine the effects of metformin, a widely-used oral anti-diabetic drug, phenformin and eight sulfenamide and sulfonamide derivatives of metformin on several haemostasis parameters. Thrombin Time (TT) tests were performed according to the available commercial method. The activity of factor X was conducted based on deficient plasma factor X. The activity of two main enzymes involved in haemostasis, thrombin and plasmin, was measured spectrophotometrically with chromogenic substrates. Protein C and antithrombin III (AT) activity assays using chromogenic substrates were conducted to determine the effect of the derivatives of metformin on these both naturally occurring anticoagulants. Two of the compounds, sulfenamide with hexyl tail and para-nitro-benzenesulfonamide significantly shortened TT. ortho-nitro sulfonamide at a concentration of 0.3-1.5 μmol/mL contributed to a significant decrease in the activity of factor X. However, sulfenamides with cyclohexyl, butyl and branched ethyl-hexyl tails at 1.5 of μmol/mL increased its activity, and simultaneously shortened PT. Additionally, ortho-nitro-benzenesulfonamide at concentrations of 1.5 μmol/mL was found to significantly decrease reaction velocity (↓ dA/dt) in the thrombin activity assay. On contrary, it was noticed that branched sulfenamide at the concentration of 1.5 μmol/mL significantly increased the enzymatic activity of plasmin. Metformin, phenformin and octyl and butyl sulfenamides were associated with a significant increase in the activity of AT. Hexyl sulfenamide and para-nitro- as well as para-trifluoro-ortho-nitro-benzenesulfonamide contributed to the decrease in the activity of protein C, while the other tested compounds did not affect its activity. In conclusion, 2-nitro-benzenesulfonamide derivative of metformin presents highly beneficial anticoagulant properties. This compound is therefore promising candidate for further in vitro and in vivo studies.
2 型糖尿病（T2DM）不仅以高血糖和胰岛素抵抗为特征，而且还以凝血和纤维蛋白溶解过程之间的平衡受损为特征。本研究旨在研究广泛使用的口服抗糖尿病药物二甲双胍、苯乙双胍以及二甲双胍的 8 个磺酰胺和磺胺衍生物对几种止血参数的影响。根据现有的商业方法进行凝血酶时间（TT）测试。根据缺乏的血浆因子 X 进行因子 X 的活性检测。使用显色底物分光光度法测量两种主要参与止血的酶，即凝血酶和纤溶酶的活性。使用显色底物进行蛋白 C 和抗凝血酶 III（AT）活性测定，以确定二甲双胍衍生物对这两种天然抗凝剂的影响。两种化合物，带有己基尾巴的磺酰胺和对硝基苯磺酰胺显著缩短 TT。邻硝基磺酰胺在 0.3-1.5μmol/mL 的浓度下有助于显著降低因子 X 的活性。然而，带有环己基、丁基和支链乙基-己基尾巴的磺酰胺在 1.5μmol/mL 时增加其活性，同时缩短 PT。此外，邻硝基苯磺酰胺在 1.5μmol/mL 的浓度下被发现显著降低凝血酶活性测定中的反应速度（↓dA/dt）。相反，注意到支链磺酰胺在 1.5μmol/mL 的浓度下显著增加纤溶酶的酶活性。二甲双胍、苯乙双胍以及辛基和丁基磺酰胺与 AT 活性的显著增加相关。己基磺酰胺和对硝基以及对三氟邻硝基苯磺酰胺有助于蛋白 C 活性的降低，而其他测试化合物则不影响其活性。总之，二甲双胍的 2-硝基苯磺酰胺衍生物具有高度有益的抗凝特性。因此，该化合物是进一步进行体外和体内研究的有前途的候选药物。
