Discovery of Putative Dual Inhibitor of Tubulin and EGFR by Phenotypic Approach on LASSBio-1586 Homologs

Abstract

Combretastatin A-4 (CA-4, 1) is an antimicrotubule agent used as a prototype for the design of several synthetic analogues with anti-tubulin activity, such as LASSBio-1586 (2). A series of branched and unbranched homologs of the lead-compound 2, and vinyl, ethinyl and benzyl analogues, were designed and synthesized. A comparison between the cytotoxic effect of these homologs and 2 on different human tumor cell lines was performed from a cell viability study using MTT with 48 h and 72 h incubations. In general, the compounds were less potent than CA-4, showing CC50 values ranging from 0.030 μM to 7.53 μM (MTT at 72 h) and 0.096 μM to 8.768 μM (MTT at 48 h). The antimitotic effect of the target compounds was demonstrated by cell cycle analysis through flow cytometry, and the cellular mechanism of cytotoxicity was determined by immunofluorescence. While the benzyl homolog 10 (LASSBio-2070) was shown to be a microtubule stabilizer, the lead-compound 2 (LASSBio-1586) and the methylated homolog 3 (LASSBio-1735) had microtubule destabilizing behavior. Molecular docking studies were performed on tubulin protein to investigate their binding mode on colchicine and taxane domain. Surprisingly, the benzyl homolog 10 was able to modulate EGFR phosphorylate activity in a phenotypic model. These data suggest LASSBio-2070 (10) as a putative dual inhibitor of tubulin and EGFR. Its binding mode with EGFR was determined by molecular docking and may be useful in lead-optimization initiatives.

Pharmacokinetic Profile Evaluation of Novel Combretastatin Derivative, LASSBio-1920, as a Promising Colorectal Anticancer Agent

“…In a previous study, LASSBio-1920 showed potent cytotoxicity and a better cytotoxic selectivity index against HL-60 cells [10][11][12]. In another study, the compound LASSBio-1586, in turn, stood out for exhibiting high cytotoxic effects against several cancer cell lines at nanomolar values (with MDA-MB435 showing the lowest IC 50 value, i.e., 64 nM), as well as the high cytotoxic selectivity index and microtubule polymerization inhibitory activity [28,29].…”

“…The study describes an N-acylhydrazone derivative of LASSBio-1586 [9], named LASSBio-1920, with potent in vitro cytotoxic activity, especially against HCT-116 cells (human colon cancer) and PC-9 cells (human lung cancer with overexpression of EGFR with L858R mutation). The study was based on QSAR (quantitative relationships between structure and activity), on in vitro analyses, and on molecular anchoring studies for the synthesis of the LASSBio-1920 analogue [10][11][12]. For that matter, molecular modeling for rEGFR in silico studies and the analysis of preliminary pharmacokinetic parameters to increase bioavailability were also carried out.…”

“…Docking studies suggested that LASSBio-1920 may be able to interact with β-tubulin through residues of DAMA-colchicine. Therefore, various potential chemical groups were bound to the molecule to achieve a strong interaction with the epidermal growth factor receptor (EGFR), a promising target for novel anticancer drugs [9][10][11][12]. However, the pharmacokinetic properties of LASSBio-1920 and if it can act using other pathways, such as inhibition of EGF receptor and/or activation of protein kinases, are unknown.…”

The Magic Methyl and Its Tricks in Drug Discovery and Development

“…Homologation [ 9 ] studies on the amide nitrogen led to the benzyl homolog of LASSBio-1586 ( 54 ), LASSBio-2070 ( 56 ), which showed microtubule-stabilizing behavior, while the methylated homolog, LASSBio-1735 ( 55 ), had microtubule-destabilizing behavior. In addition, none of the compounds had better cytotoxic activity when compared to the N -methylated compound LASSBio-1735 ( 55 ) [ 46 ].…”

“…Barbosa and colleagues [46] described a series of combretastatin A-4 analogs based on the N-acylhydrazone (NAH) LASSBio-1586 (54) with cytotoxic and antimitotic activity (Figure 14). Homologation [9] studies on the amide nitrogen led to the benzyl homolog of LASSBio-1586 (54), LASSBio-2070 (56), which showed microtubule-stabilizing behavior, while the methylated homolog, LASSBio-1735 (55), had microtubule-destabilizing behavior.…”