3,3'-((3-Hydroxyphenyl)azanediyl)dipropionic Acid Derivatives as a Promising Scaffold Against Drug-Resistant Pathogens and Chemotherapy-Resistant Cancer

Abstract

The synthesis and antimicrobial and anticancer activity of 3,3'-((3-hydroxyphenyl)azanediyl)dipropionic acid derivatives (2-25) against drug-resistant bacterial pathogens and FaDu head and neck cancer cells were investigated. The derivatives were synthesized through various methods, including esterification, hydrazinolysis, and condensation reactions. The compounds demonstrated structure-dependent antimicrobial activity, predominantly targeting Gram-positive pathogens. Compounds containing 4-nitrophenyl, 1-naphthyl, and 5-nitro-2-thienyl groups exhibited enhanced activity against S. aureus and E. faecalis. Additionally, compounds 5, 6, and 25 showed antiproliferative activity in cisplatin-resistant FaDu cells at low micromolar concentrations. The in silico modeling revealed that compound 25 interacts with the HER-2 and c-MET proteins. These compounds also induced significant oxidative stress in FaDu cells and demonstrated low cytotoxic activity in non-cancerous HEK293 cells. These results highlight the potential of N-aryl-substituted β-amino acid derivatives as promising scaffolds for the further development of novel amino acid-based antimicrobial and anticancer agents targeting drug-resistant pathogens and cancers.

Keywords: 3,3′-((3-hydroxyphenyl)azanediyl)dipropionic acid; HER2; antimicrobial activity; c-MET; head and neck cancer; multidrug-resistant pathogens; β-amino acids derivatives.

Figure 1

Derivatives of β-amino acids.

Scheme 1

Synthesis of compounds 2–7. (I) acrylic acid, H₂O, 75 °C, 5 h; (II) CH₃OH, H⁺, reflux, 7 h; (III) N₂H₄.H₂O, i-PrOH, reflux, 6 h; (IV) carbamide, HCl, reflux, 1 h; (V) KSCN, HCl, reflux, 1 h; (VI) acrylic acid, hydroquinone, C₆H₆, reflux, 3 h.

Scheme 2

Synthesis of hydrazones 8–22. (VII) ArCHO, i-PrOH, reflux, 2 h; (VIII) HetCHO, i-PrOH, reflux, 2 h; (IX) CH₃COR, CH₃OH (22), reflux, 5 h. 8, Ar=C₆H₅; 9, Ar=2,4-F₂-C₆H₃; 10, Ar=4-NO₂-C₆H₄; 11, Ar=4-Cl-C₆H₄; 12, Ar=4-(CH₃)₂N-C₆H₄; 13, Ar=3,4,5-(OCH₃)₃-C₆H₂; 14, Ar=1-naphthyl; 15, Het=2-furyl; 16, Het=5-nitro-2-thienyl; 17, Het=5-nitro-2-furyl; 18, Het=3-thienyl; 19, R=CH₃; 20, R=C₂H₅; 21, R=C₆H₅; 22, R=4-H₂NO₂SC₆H₄.

Scheme 3

Synthesis of compounds 23–25. (X) hexan-2,5-dione, CH₃COOH, i-PrOH, reflux, 5 h; (XI) pentane-2,4-dione, HCl, i-PrOH, reflux, 5 h; (XII) isatin, CH₃COOH, i-PrOH, reflux, 12 h.

Figure 2

The 3,3′-((3-hydroxyphenyl)azanediyl)dipropionic acid derivatives (2–25) demonstrate structure-dependent anticancer activity in FaDu cells. The cells were exposed to the compounds, cisplatin (CP), or doxorubicin (DOX) for 24 h, and viability was measured by using the MTT assay. Data are presented as means ± SD from three experimental replicates. Statistical significance was determined by using a one-way ANOVA test. **—p < 0.01; ***—p < 0.001.

Figure 3

The selected 3,3′-((3-hydroxyphenyl)azanediyl)dipropionic acid derivatives (5, 6, and 25) exhibit favorable cytotoxic effects in non-cancerous HEK293T cells. (A) shows dose–response kinetics of compounds 5, 6, 25, and control pharmaceuticals in HEK293 cells, while (B) demonstrates the calculated IC₅₀ values. Cells were treated with the compounds, cisplatin (CP), or doxorubicin (DOX) for 24 h, and viability was assessed by using the MTT assay. Data are presented as means ± SD from three independent experiments.

Figure 4

The most promising 3,3′-((3-hydroxyphenyl)azanediyl)dipropionic acid derivatives, 5, 6, and 25, show dose-dependent anticancer activity in FaDu cells. (A) demonstrates the dose–response kinetics of these compounds, as well as cisplatin (CP) and doxorubicin (DOX), in FaDu cells. (B) shows the IC₅₀ kinetics of these compounds in FaDu cells.

Figure 5

Compounds 5, 6, and 25 induce oxidative stress in FaDu cells. (A) demonstrates treatment-induced SOD activity, expressed as U/mL of cell lysate, while (B) shows relative hydrogen peroxide formation, expressed as optical density at 525 nm. The FaDu cells were exposed to a fixed concentration of compounds and test drugs (cisplatin (CP) and doxorubicin (DOX)) for 6 h in medium containing 0.25% DMSO, after which SOD activity and hydrogen peroxide levels were quantified by using commercial kits. The data are expressed as means ± SD from three experimental replicates for SOD activity or mean ± SD from six experimental replicates for hydrogen peroxide quantification. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; ns—not significant.

Figure 6

Compound 25 is proposed to interact with human epidermal growth factor receptor 2 (HER2) based on in silico molecular modeling. (A) visualizes the potential binding site of compound 25 (green), the ligand (silver), and erlotinib (yellow) within HER2. (B) presents 2D interaction maps illustrating hydrogen bonds (H-bonds) and hydrophobic interactions between compound 25 and HER2 residues. Van der Waals and π interactions are categorized as hydrophobic interactions.

Figure 7

Compound 25 is proposed to interact with human mesenchymal–epithelial transition factor (c-Met) based on in silico molecular modeling. (A) visualizes the potential binding site of compound 25 (green), the ligand (silver), and erlotinib (yellow) within c-Met. (B) presents 2D interaction maps illustrating hydrogen bonds (H-bonds) and hydrophobic interactions between compound 25 and c-Met residues. Van der Waals and π interactions are categorized as hydrophobic interactions.