In vitro anticancer potentiality and molecular modelling study of novel amino acid derivatives based on N¹,N³-bis-(1-hydrazinyl-1-oxopropan-2-yl) isophthalamide

Figures & data

Figure 1. Design of newly synthesized isophthalamide based derivatives 4–16 concerning the chemical structures of poly (l-lysine isophthalamide) (I) and different heterocyclic motifs II–IX with various mechanisms of anticancer activity.

Scheme 1. Synthetic routes for N¹,N³-bis-(1-hydrazinyl-1-oxopropan-2-yl) isophthalamide (4).

Scheme 2. Synthetic routes for N¹,N³- bis-(1-hydrazinyl-1-oxopropan-2-yl) isophthalamide derivatives 5–9.

Scheme 3. Synthetic routes for N¹,N³-bis-(1-hydrazinyl-1-oxopropan-2-yl) isophthalamide derivatives 10–16.

Figure 2. In vitro cytotoxicity of compounds 4–16, against human colon tumour (HCT-116), lung tumor (A-549) and human breast tumour (MCF-7) cell lines at concentration 100 µM. Each result is a mean of 3 replicate and values are represented as % inhibition (± standard error).

Table 1. IC₅₀ values (concentration required to diminish 50% of the cell) of active compounds possessing ≥ 70% cytotoxicity on colon carcinoma (HCT-116), lung carcinoma (A-549) and breast carcinoma (MCF-7) cell lines.

	IC50 (mean ± SD) (µM)^a
Compd. no.	HCT-116	A-549	MCF-7
5	0.014 ± 0.22	0.040 ± 1.43	0.014 ± 0.70
7	–	–	0.073 ± 2.91
12	–	0.061 ± 2.12	0.051 ± 3.10
14	0.054 ± 1.80	0.041 ± 1.91	0.031 ± 1.51
Doxorubicin	0.065 ± 1.00	0.049 ± 1.30	0.045 ± 2.20

^aValues of IC (±standard error) are calculated using SPSS statistical program.

Figure 3. cytotoxic percentage upon normal skin human cell line (BJ-1) at concentration 100 µM of those compounds gave ≥ 70% cytotoxicity over the three tumour cell lines. Each result is a mean of 3 replicate and values are represented as % inhibition (± standard error).

Table 2. Inhibitory evaluation of compound 5 against EGFR, VEGFR-2, CDK-2 and c-Kit kinases.

Kinase	IC50 (mean ± SD) (µM)
	5	Staurosporine
EGFR	0.78 ± 1.25	1.00 ± 1.12
VEGFR-2	3.17 ± 1.00	0.08 ± 1.10
CDK-2	6.24 ± 1.21	0.11 ± 1.13
c-kit	2.85 ± 0.04	0.60 ± 1.00

IC₅₀: compound concentration required to inhibit the enzyme activity by 50%, SEM = standard error mean; each value is the mean of three values.

Figure 4. quantitative analysis of Bax, BCL-2 and Bax/BCL-2 ratio and p53 for compound 5 compared to doxorubicin, where Y-axis represents the protein levels (ng/ml).

Figure 5. Graphical representation for Caspase-7 concentration (ng/ml) in treated breast cancer cells with compound 5 and untreated compared to doxorubicin and for % DNA fragmentation and IC₅₀ (ng/ml) on tubuline B in comparison with colchicine.

Figure 6. (A and B) Images display 2D and 3D graphs of compound 5 docked into EGFR binding sit (PDB code: 1M17). Green colour indicates hydrophobic area, pink colour indicates high polar area, blue colour indicates mild polar area and dotted lines and arrows represent hydrogen bonds.