Structural Comparison of Sulfonamide-Based Derivatives That Can Improve Anti-Coagulation Properties of Metformin

Figures

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  Figure 1. Structure of examined biguanides. Explanatory notes: Ac—acetyl, CN—cyano, Cy—cyclohexyl, Ph—phenyl.

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  Figure 2. Effects of metformin and biguanide analogues 1–5 at the concentration of 0.1 (A) and 0.3 mM (B), and D-glucose at the concentration of 0.1 mM on the 2-NBDG uptake in HUVEC cells. The results are presented as mean ± SD, n = 4–6, and an asterisk denotes the significant differences between control and cells treated with glucose, metformin, or studied compounds. * p < 0.05; ** p < 0.01; *** p < 0.001.

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  Figure 3. Effect of compounds 1–8 on HUVEC and AoSMC viability and morphology after 24-h incubation. Cells were cultured without (control, CTR) and in the presence of compounds 1–8 at a concentration of 0.006–5.0 mM. Representative cell images are shown for concentrations corresponding to IC50 values or 3 mM (100-fold magnification). Metformin was found to not affect the viability and morphology of HUVEC and AoSMC cells in our previous study [28].

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  Figure 4. The effects of compounds 1–5 on the integrity of HUVEC analyzed in the RTCA-DP system. (A) The effect of the exposure of compounds 1–5 on normalized Cell Index (nCI) at selected time points (3, 6, 12, 24, and 36 h). The results are presented as mean ± SD, n = 4–5. An asterisk denotes a significant difference between sample treated with biguanide and control sample; * p < 0.05; ** p < 0.01; *** p < 0.001. (B) The effects of compound 5 on the barrier properties of HUVECs. The picture presents representative plots of one experiment conducted in duplicates (the results are presented as a mean (solid line) ± standard deviation). For the statistical analysis, three independent experiments were conducted. Red line—control (unstimulated cells); green line—compounds at the concentration of 0.3 mM; navy blue line—1.0 mM.

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  Figure 5. Impact of selected sulfonamides 1–5 and metformin on the migration of HUVECs was analyzed using wound-healing assay in the JulieStage system. The represented graphs reflect changes of the wound width (µm) during 24 h in the absence (control) and in the presence of selected compounds at the concentration of 0.3 (A) and 1.0 mM (B). The results are presented as mean ± SD, n = 4–8; (* p < 0.05).

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  Figure 6. Effects of biguanides 1–5 on the activity of factor X (data are presented as mean ± SD; n = 4–5) after 1-min incubation with plasma-deficient factor X. The asterisk denotes a statistically significant difference between the samples treated with compounds and control; * p < 0.05, ** p < 0.01.

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  Figure 7. Effects of selected biguanides (1–5) on the activity of anti-thrombin III (AT) (data are presented as mean ± SD; n = 4). The asterisk denotes a statistically significant difference between the samples treated with compounds and control; * p < 0.05, ** p < 0.01.

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  Figure 8. Influence of chosen biguanides 1–5 on overall potential of clot formation and fibrinolysis (CLAUC), and selected parameters of clot formation and fibrinolysis process: maximum clotting (Fmax); initial plasma clotting velocity (Fvo); initial clot fibrinolysis velocity (Lvo) (data are presented as means, n = 5–7). The asterisk denotes a statistically significant difference between the samples treated with compounds and respective controls; * p < 0.05, ** p < 0.01, *** p < 0.001. In our previous study, metformin did not affect the overall potential of clot formation and fibrinolysis (CLAUC) over the entire concentration range and dimethyl biguanide did not influence the kinetic parameters of the clot formation process and fibrinolysis (Fmax, Fvo, Lvo constants) [37].