Cancer treatment remains a serious challenge worldwide. Thus, finding novel antitumour agents is of great importance. In the present study, nine new benzenesulphonohydrazide derivatives (
In 2018, >1.7 million cancer deaths worldwide were caused by lung cancer that remains the most common cancer (11.6% of all cancers) and the main cause of cancer-related deaths (
Despite the advances in cancer treatment, liver and kidney tumours are also associated with high fatality rates (
Renal cell carcinoma (RCC), which accounts for 90% of kidney cancer cases, is the most lethal among urological tumours (
Considering the dismal prognosis of patients diagnosed with metastatic RCC, advanced SCC or liver cancer, developing novel drug candidate molecules is important to improve the outcomes in these diseases. Thus, the present study focused on new benzenesulphonohydrazide derivatives. Benzenesulphonohydrazide derivatives have been reported to exert various biological activities, including antitumour (
The present study aimed to design, synthesize and analyse new benzenesulphonohydrazones to identify potential novel antitumour agents for RCC, SCC and liver cancer.
The reagents and solvents used in the present study were purchased from Sigma-Aldrich; Merck KGaA and were used without further purification. The melting points were determined using the Fisher-Johns blocks melting point apparatus (Thermo Fisher Scientific, Inc.) and left uncorrected. Fourier transform infrared spectra were recorded on a Nicolet 6700 spectrometer (Thermo Fisher Scientific, Inc.), ν in cm−1. 1H nuclear magnetic resonance (NMR) and 13C NMR spectra were recorded on a Bruker Avance 300 apparatus (Bruker Corporation) in DMSO-
4-Methylbenzenesulphonohydrazide (0.01 mol) was dissolved in 10 ml 96% ethanol, and an appropriate substituted aromatic aldehyde (0.011 mol) was added. The mixture was heated under reflux for 3 h. Then, the solution was allowed to cool down. Subsequently, the formed precipitate was filtered off under low pressure and recrystallized from ethanol. Detailed physico-chemical information of the synthesized benzenesulphonohydrazones 1–9 is presented in Supplementary Materials and methods (
The experimental lipophilicity of the synthesized benzenesulphonohydrazide derivatives 1–9 (
The present study used tumour cell cultures supplied by the American Type Culture Collection (ATCC), including 769-P (human renal cell adenocarcinoma; ATCC® CRL-1933™), HepG2 (human liver cancer; ATCC® HB-8065™), NCI-H2170 (H2170; human squamous cell carcinoma of the lung; ATCC® CRL-5928™), as well as Vero cells (monkey kidney; ATCC® CCL-81™), which was used as a normal reference cell line in similar biological activity-focused studies (
The inhibitory effects of compounds 1–9 on cell proliferation were assessed by MTT assay [DB-ALM Protocol no. 17: MTT Assay; European Union Reference Laboratory, European Center for the Validation of Alternative Methods (EURL ECVAM), Database Service on Alternative Methods to Animal Experimentation (DB-ALM);
New benzenesulphonylhydrazones 1–9 were obtained based on the condensation reaction of 4-methylbenzenesulphonohydrazide with appropriate aldehydes (
The highest lipophilicity value was presented by compound 4, which contained a 4-bromo-2-fluorophenyl substituent, irrespective of the organic modifier used (
Based on our previous studies (
Assessment of the
Based on the increasing incidence and mortality rates of cancer, patients with cancer still need new drugs with high efficacy and selectivity to minimize side effects. Hydrazide-hydrazones and benzenesulphonohydrazones exhibit anticancer activity (
The present study was a pilot study focused on the synthesis, lipophilicity and screening of the antiproliferative potential of newly developed benzenesulphonohydrazones. The results of the antitumour activity analysis demonstrated that the substitution with fluorine at the
Among the tested molecules, compound 7 was the most cytotoxic and selective in H2170 cells. However, when comparing the IC50 values estimated for the above compound with those of cisplatin, the activity of compound 7 appeared weak (
The results of the present study demonstrated that compounds 2, 6 and 7 were cytotoxic and selective to cancer cells, in particular for renal cell adenocarcinoma. These compounds were substituted at the phenyl ring with chlorine or bromine atoms at the
In summary, the results of the present study demonstrated that the newly synthesized benzenesulphonohydrazones 1–9 exhibited antitumour potential, particularly compound 4, which was characterized by high, selective and diverse cytotoxicity to the tested cancer cell lines, whereas in the normal reference cells, its IC50 was determined to be higher. The compounds that were substituted with a halogen and methoxy group in the phenyl ring (compounds 2, 6 and 7) effectively inhibited the viability of 769-P cells and, to a lesser extent, that of HepG2 and H2170 cells, while exerting a limited effect on Vero cells. Across all compounds, the combination of fluorine and bromine substituents at the phenyl ring appeared to be the most advantageous, providing high cytotoxicity and selectivity towards tumour cells. Renal adenocarcinoma cells were the most sensitive to the antiproliferative potential of the newly developed molecules. Although this is a preliminary study, the obtained results suggested the direction of further research including the synthesis of new compounds of this chemical group. Even the preliminary results of the pilot attempt are relevant in the context of planning the synthesis of novel derivatives for pre-clinical studies. In addition, in our future work, other cancer lines will be tested as well as the mechanisms of cytotoxic activity of the most promising newly synthesized derivatives in order to examine and explain its antiproliferative potential. Our future studies will include the evaluation of apoptosis through various techniques such as the Annexin V, western blot or Caspase Glo assays. Additionally, antiproliferative mechanism-focused experiments will also comprise the assessment of the effects of the new compounds on the cell cycle. Revealing the mechanisms responsible for the cytotoxic effect of the newly synthesized molecules may facilitate the development of new effective anticancer therapies.
Not applicable.
No funding was received.
All the data generated and/or analysed during this study are included in this published article.
ŁP designed the study, performed the synthesis of the benzenesulphonohydrazide derivatives, analysed the spectral data and wrote the first draft of the manuscript, with the exception of the lipophilicity and cytotoxicity sections. MGG, IPC, DNC and MH performed the cytotoxicity assays and wrote the cytotoxicity section of the manuscript. ABR performed the lipophilicity analysis of the obtained compounds and wrote the associated section of the manuscript. KP assisted with the synthesis of the benzenesulphonohydrazide derivatives. AG supervised and was involved in the conception of the lipophilicity analyses, helped with interpretation of lipophilicity data of synthesized compounds and revised critically section devoted to lipophilicity study in the final version of manuscript. JD supervised and was involved in the conception of the cytotoxicity analyses, helped with interpretation of cytotoxicity data of obtained derivatives and revised critically section devoted to cytotoxicity study in the final version of manuscript. MW supervised the synthesis and helped with the interpretation of the spectral data of the synthesized compounds. All authors read and approved the final manuscript.
Not applicable.
Not applicable.
The authors declare that they have no competing interests.
Synthesis of new benzenesulphonohydrazide derivatives. Compound 1: R = 2,3-diF; Compound 2: R = 2-Cl-3-OMe; Compound 3: R = 2-Br-3-OH-4-OMe; Compound 4: R = 2-F-4-Br; Compound 5: 2,3-diOMe; Compound 6: R = 3-Cl-4-OMe; Compound 7: R = 3-Br-4-OMe; Compound 8: R = 3-Br-4-OH; Compound 9: R = 3-OEt-4-OH.
Effects of substituents on the antitumour activity of the synthesized benzenesulphonohydrazones.
Log PEXP values of the synthesized compounds calculated by the standardization method in different solvents.
Compound no. | R | log Pacetone | log Pacetonitrile | log P1,4-dioxane | log Pmethanol |
---|---|---|---|---|---|
1 | 2,3-diF | 2.06 | 3.20 | 4.38 | 3.00 |
2 | 2-Cl-3-OMe | 3.33 | 4.31 | 3.96 | 5.56 |
3 | 2-Br-3-OH-4-OMe | 3.74 | 4.02 | 3.86 | 5.93 |
4 | 2-F-4-Br | 4.38 | 4.11 | 4.81 | 6.56 |
5 | 2,3-diOMe | 2.94 | 3.53 | 3.46 | 4.55 |
6 | 3-Cl-4-OMe | 3.43 | 3.51 | 3.76 | 5.27 |
7 | 3-Br-4-OMe | 3.69 | 3.63 | 3.94 | 5.39 |
8 | 3-Br-4-OH | 2.23 | 3.50 | 2.26 | 2.35 |
9 | 3-OEt-4-OH | 2.17 | 3.96 | 2.21 | 2.24 |
R, substituent; log PEXP, log P values experimentally obtained using thin layer chromatography method.
Antiproliferative activity and selectivity of the newly synthesized benzenesulphonohydrazide derivatives.
Compound no. | R | IC50 in Vero, µM | IC50 in 769-P, µM | Selectivity index |
IC50 in HepG2, µM | Selectivity index |
IC50 in H2170, µM | Selectiviy index |
---|---|---|---|---|---|---|---|---|
1 | 2,3-diF | 259.00 | >500.00 | nd | >500.00 | nd | >500.00 | nd |
2 | 2-Cl-3-OMe | 1,646.34 | 67.56 | 24.37 | 160.46 | 10.26 | 501.55 | 3.28 |
3 | 2-Br-3-OH-4-OMe | 396.24 | 144.29 | 2.75 | 200.08 | 1.98 | 387.24 | 1.02 |
4 | 2-F-4-Br | 964.69 | 73.62 | 13.10 | 106.33 | 9.07 | 369.88 | 2.61 |
5 | 2,3-diOMe | 108.93 | 1.94 | 56.15 | >500.00 | nd | >500.00 | nd |
6 | 3-Cl-4-OMe | 1,457.83 | 26.38 | 55.26 | 364.94 | 3.99 | 517.37 | 2.82 |
7 | 3-Br-4-OMe | 1,208.49 | 100.63 | 12.00 | >500.00 | nd | 314.55 | 3.84 |
8 | 3-Br-4-OH | 157.20 | 74.54 | 2.11 | nd | nd | >500.00 | nd |
9 | 3-OEt-4-OH | 146.63 | 54.23 | 2.70 | 285.95 | 0.51 | >500.00 | nd |
R, substituent; nd, not determined.
Calculated as the ratio between the IC50 values in the non-tumour cell line Vero and the respective cancer cell lines.