Study of reactivity of cyanoacetohydrazonoethyl-N-ethyl-N-methyl benzenesulfonamide: preparation of novel anticancer and antimicrobial active heterocyclic benzenesulfonamide derivatives and their molecular docking against dihydrofolate reductase

Figures & data

Table 1. In vitro anticancer screening of the synthesized compounds against human breast cell line (MCF-7).

	Compound concentration (μg/mL)
Cpd No	0.39	0.78	1.56	3.125	6.25	12.5	25	50	IC50*(μg/mL)
2	100 ± 0.0	100 ± 0.0	98.58 ± 0.6	94.43 ± 0.8	87.69 ± 1.5	76.14 ± 4.3	32.19 ± 1.9	14.48 ± 1.1	19.9
3	100 ± 0.0	100 ± 0.0	97.41 ± 0.6	91.38 ± 1.1	85.65 ± 1.5	62.19 ± 4.2	23.61 ± 1.7	10.24 ± 0.4	16.4
4	100 ± 0.0	100 ± 0.0	93.53 ± 0.6	87.82 ± 1.5	70.19 ± 4.6	58.65 ± 3.4	18.27 ± 1.5	7.43 ± 03	4.2
5	100 ± 0.0	100 ± 0.0	100 ± 0.0	98.43 ± 0.6	92.05 ± 0.8	83.17 ± 1.5	35.42 ± 2.7	26.81 ± 1.4	21.2
6	100 ± 0.0	100 ± 0.0	90.31 ± 0.9	79.54 ± 1.5	62.48 ± 3.7	24.96 ± 1.9	11.72 ± 1.0	5.87 ± 0.2	8.33
7	100 ± 0.0	100 ± 0.0	94.91 ± 0.8	89.12 ± 1.2	67.65 ± 4.6	49.18 ± 3.3	28.96 ± 1.7	11.23 ± 0.7	12.2
8	64.4 ± 4.6	56.82 ± 3.9	29.35 ± 2.8	26.84 ± 1.7	14.61 ± 1.1	9.45 ± 1.0	7.38 ± 0.3	3.97 ± 0.2	1.08
9	100 ± 0.0	100 ± 0.0	98.36 ± 0.6	94.65 ± 0.8	86.82 ± 1.1	75.23 ± 1.5	37.38 ± 1.9	10.89 ± 1.0	20.8
10	100 ± 0.0	100 ± 0.0	98.11 ± 0.6	94.76 ± 0.8	88.18 ± 1.2	67.82 ± 4.5	16.27 ± 1.2	7.19 ± 0.5	16.8
11	100 ± 0.0	100 ± 0.0	100 ± 0.0	97.82 ± 0.6	91.54 ± 0.9	78.71 ± 1.5	43.68 ± 3.2	27.92 ± 1.8	22.7
12	100 ± 0.0	100 ± 0.0	94.25 ± 0.7	87.42 ± 1.1	78.36 ± 1.5	61.59 ± 4.6	18.37 ± 1.5	10.94 ± 0.6	4.9
13	100 ± 0.0	100 ± 0.0	98.97 ± 0.6	96.28 ± 0.7	89.31 ± 1.1	77.14 ± 1.5	46.55 ± 3.2	28.72 ± 1.7	23.6
14	100 ± 0.0	100 ± 0.0	98.44 ± 0.6	92.85 ± 0.9	84.93 ± 1.5	68.72 ± 4.6	32.89 ± 1.8	20.64 ± 1.2	19
15	100 ± 0.0	100 ± 0.0	100 ± 0.0	97.89 ± 0.6	91.37 ± 1.1	79.55 ± 1.5	27.18 ± 1.7	14.67 ± 1.1	19.6
MTX	100 ± 0.0	100 ± 0.0	95.32 ± 0.6	89.31 ± 0.8	67.70 ± 0.6	50.12 ± 1.3	29.71 ± 1.5	11.56 ± 0.6	12.3

*Mean of three results obtained from three experiments.

IC₅₀ value: concentration causing 50% inhibition of cell viability.

Table 2. In vitro cytotoxicity screening of the synthesized compounds against normal fibroblasts of baby hamster kidney cell line (BHK).

	Compound concentration (μg/mL)
Cpd No	1.56	3.125	6.25	12.5	25	50	100	200	CC50*(μg/mL)
2	100 ± 0.0	98.13 ± 0.6	94.82 ± 0.8	90.76 ± 1.1	86.38 ± 1.4	71.89 ± 1.5	63.24 ± 3.3	41.06 ± 3.1	160
3	100 ± 0.0	100 ± 0.0	98.71 ± 0.6	94.32 ± 0.9	87.51 ± 1.1	71.24 ± 1.5	52.98 ± 3.8	34.86 ± 1.9	116
4	100 ± 0.0	99.16 ± 0.0	97.24 ± 0.6	91.43 ± 1.2	82.16 ± 1.5	68.74 ± 4.6	50.31 ± 4.1	31.48 ± 2.7	102
5	100 ± 0.0	100 ± 0.0	98.95 ± 0.6	97.83 ± 0.8	91.42 ± 0.9	82.39 ± 1.3	64.95 ± 4.6	48.72 ± 3.1	192
6	98.62 ± 0.6	96.21 ± 0.8	92.87 ± 1.1	81.44 ± 1.4	70.36 ± 4.6	49.21 ± 3.3	37.49 ± 2.7	23.75 ± 1.9	49.1
7	100 ± 0.0	98.74 ± 0.6	95.20 ± 0.9	88.46 ± 1.2	79.21 ± 1.5	65.23 ± 4.6	46.89 ± 3.9	30.46 ± 1.8	91.5
8	96.97 ± 0.7	94.32 ± 0.9	87.21 ± 1.1	78.14 ± 1.4	69.28 ± 4.5	54.13 ± 3.9	39.42 ± 2.8	26.89 ± 1.7	64
9	100 ± 0.0	100 ± 0.0	98.93 ± 0.6	96.12 ± 0.8	87.53 ± 1.1	73.84 ± 1.5	48.62 ± 3.6	31.46 ± 1.9	97.3
10	100 ± 0.0	99.84 ± 0.0	95.86 ± 0.9	90.43 ± 1.3	78.92 ± 1.5	51.38 ± 4.1	39.24 ± 2.7	20.98 ± 1.5	55.7
11	100 ± 0.0	100 ± 0.0	100 ± 0.0	99.27 ± 0.0	94.18 ± 0.9	87.24 ± 1.3	71.43 ± 1.5	53.89 ± 3.9	>200
12	100 ± 0.0	100 ± 0.0	97.13 ± 0.6	90.28 ± 1.1	78.34 ± 1.5	65.26 ± 4.4	44.19 ± 3.2	26.86 ± 1.7	86.2
13	100 ± 0.0	100 ± 0.0	100 ± 0.0	97.28 ± 0.6	91.43 ± 0.9	86.22 ± 1.1	78.19 ± 1.5	60.38 ± 4.6	>200
14	100 ± 0.0	100 ± 0.0	100 ± 0.0	98.29 ± 0.6	90.76 ± 0.1	78.42 ± 1.5	60.87 ± 4.2	41.23 ± 3.1	45
15	100 ± 0.0	99.17 ± 0.0	95.22 ± 0.7	86.91 ± 1.1	74.86 ± 1.5	60.98 ± 4.1	47.51 ± 3.7	36.17 ± 1.9	90.8
MTX	100 ± 0.0	100 ± 0.0	99.10 ± 0.9	93.75 ± 1.2	83.92 ± 1.5	70.14 ± 1.3	52.41 ± 1.5	33.21 ± 1.5	105.2

*Mean of three results obtained from three experiments.

CC₅₀ value: concentration required to inhibit cell proliferation by 50%.

Table 3. CC₅₀, IC₅₀ (in μg/mL and μM) and selectivity index SI of the synthesized compounds against breast carcinoma cell line (MCF-7).

Cpd No	CC50* (μg/mL)	CC50* (μM)	IC50* (μg/mL)	IC50* (μM)	SI
2	160	496.89	19.9	61.8	8.04
3	116	276.19	16.4	39.05	7.07
4	102	218.42	4.2	32.55	6.71
5	192	457.14	21.2	50.48	9.06
6	49.1	113.13	8.33	19.19	5.89
7	91.5	187.12	12.2	24.95	7.5
8	64	149.88	1.08	2.53	59.26
9	97.3	203.98	20.8	43.61	4.68
10	55.7	142.82	16.8	43.08	3.32
11	>200	550.96	22.7	62.53	8.81
12	86.2	42.57	4.9	28.14	5.42
13	>200	406.5	23.6	47.97	8.47
14	45	461.31	19	56.55	8.16
15	90.8	214.4	19.6	46.23	4.63
MTX	105.2	231.72	12.3	27.09	8.55

*Mean of three results obtained from three experiments.

CC₅₀ value: concentration required to inhibit cell proliferation by 50%.

IC₅₀ value: concentration causing 50% inhibition of cell viability.

SI = CC₅₀/IC₅₀ selectivity index.

Table 4. Anti-fungal activity of synthesized compounds*.

	Microorganisms
Cpd No	Aspergillus fumigatus(RCMB 02568)	Candida albicans(RCMB 05036)
2	NA†	NA
3	NA	NA
4	18.3 ± 0.25	NA
5	4.6 ± 0.10	NA
6	NA	NA
7	20.3 ± 0.24	NA
8	17.6 ± 0.58	NA
9	NA	NA
10	17.9 ± 0.36	NA
11	NA	NA
12	20.2 ± 0.20	NA
13	NA	NA
14	17.4 ± 0.16	NA
15	17.6 ± 0.58	NA
Amphotericin B	23.7 ± 0.10	21.9 ± 0.12

*Mean zone of inhibition in mm ± standard deviation (SD); mm: the unit of measurement of inhibition zone (IZ); †NA: No activity. The test was performed using the diffusion agar technique.

Table 5. Anti-bacterial activity of synthesized compounds*.

	Microorganisms
	Gram-positive bacteria			Gram-negative bacteria
Cpd. No	Staphylococcus aureus(RCMB 010027)	Streptococcus pneumoniae(RCMB 010010)	Bacillis subtilis(RCMB 010067)	Pseudomonas aeruginosa(RCMB 010043)	Klebsiella pneumoniae(RCMB 010093)	Escherichia coli(RCMB 010052)
2	NA†	NA	NA	NA	NA	NA
3	NA	NA	NA	NA	NA	NA
4	16.2 ± 0.34	18.3 ± 0.43	20.3 ± 0.53	NA	20.3 ± 0.25	17.8 ± 0.43
5	12.6 ± 0.18	14.8 ± 0.16	16.4 ± 0.12	NA	18.1 ± 0.4	4.2 ± 0.30
6	NA	NA	NA	NA	NA	NA
7	18.7 ± 0.62	19.5 ± 0.42	20.2 ± 0.29	NA	20.9 ± 0.28	19.3 ± 0.13
8	4.4 ± 0.25	16.2 ± 0.4	19.8 ± 0.42	NA	17.4 ± 0.53	4.0 ± 0.43
9	NA	NA	NA	NA	NA	NA
10	4.9 ± 0.44	18.1 ± 0.63	20.0 ± 0.32	NA	NA	13.9 ± 0.46
11	NA	NA	NA	NA	NA	NA
12	19.3 ± 0.16	20.6 ± 0.23	22.8 ± 0.18	NA	21.2 ± 0.29	19.4 ± 0.10
13	NA	NA	NA	NA	NA	NA
14	4.2 ± 0.34	16.1 ± 0.31	18.1 ± 0.25	NA	19.6 ± 0.31	17.5 ± 0.22
15	4.4 ± 0.25	17.7 ± 0.43	19.3 ± 0.53	NA	13.7 ± 0.25	NA
Ampicillin	28.9 ± 0.14	25.3 ± 0.58	26.4 ± 0.34	–	–	–
Gentamicin	–	–	–	26.3 ± 0.4	17.3 ± 0.12	27.3 ± 0.44

*Mean zone of inhibition in mm ± standard deviation (SD); mm: the unit of measurement of inhibition zone (IZ); †NA: No activity. The test was performed using the diffusion agar technique.

Scheme 1. Synthesis of starting material 4-(1-(2-(2-cyanoacetyl) hydrazono)ethyl)-N-ethyl-N-methylbenzenesulfonamide (2).

Scheme 2. Synthesis of thiophenes 3, 4, 5 and 6, thiazole 7 and chromenes 8 and 9 derivatives.

Scheme 3. Synthesis of piperidine 10, pyrrolidine 11, tetrahydropyridine 12, pyridine 13, pyrazole 14 and hydrazone 15 derivatives.

Figure 1. Ligand–protein interactions in methotrexate-DHFR X-ray complex A) 3D, B) 2D interactions.

Table 6. Docking score energy of the selective newly synthesized compounds.

Cpd. No	Score	E-conf	E-place	E-score 1	E-score 2	E-refine
2	−20.4574	3.596101	−75.0351	−10.9972	−20.4574	−19.8169
3	−21.4633	41.8881	−109.539	−11.496	−21.4633	−23.3608
4	−24.2934	42.20723	−119.642	−12.193	−24.2934	9.963194
5	−22.1118	42.98828	−81.014	−11.3328	−22.1118	−25.7723
6	−23.0272	30.60611	−108.314	−11.1684	−23.0272	−21.0398
7	−23.4163	73.64976	−98.7849	−12.0301	−23.4163	−4.59358
8	−22.403	52.45129	−112.641	−11.432	−22.403	−17.8918
9	−25.7524	67.21304	−97.5068	−11.616	−25.7524	2.963847
10	−21.8549	−4.1024	−90.8791	−11.5605	−21.8549	−8.5729
11	−21.6592	5.367175	−74.9848	−11.7363	−21.6592	−13.4769
12	−28.6922	73.42744	−103.767	−10.7284	−28.6922	7.603124
13	−24.1631	52.55446	−92.257	−11.5454	−24.1631	−4.2026
14	−20.1099	−5.04918	−87.9892	−10.2547	−20.1099	−13.9655
15	−23.5028	46.12006	−76.1009	−11.4817	−23.5028	−14.7353
MTX	−28.2736	143.6868	−125.016	−12.3235	−28.2736	39.20521