Coculture-Based Screening Revealed Selective Cytostatic Effects of Pyrazol-Azepinoindoles.

Area of Science:

• Drug Discovery and Development
• Medicinal Chemistry
• Cancer Biology

Background:

• The search for novel anticancer agents is crucial for effective cancer therapy.
• Early-stage screening methods are vital for identifying compounds with specific cytotoxic activity.
• Existing drug discovery pipelines require innovative approaches to identify new molecular entities.

Purpose of the Study:

• To identify novel small molecules with selective anticancer activity.
• To utilize the fluorescent cells cocultivation test (FCCT) for early-stage drug development.
• To investigate structure-activity relationships of promising anticancer compound scaffolds.

Main Methods:

• Screening of 2025 compounds using the fluorescent cells cocultivation test (FCCT) with breast cancer (MCF7) and noncancerous (MCF10A) cells.
• Confirmation of cytotoxicity using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.
• Structure-activity relationship (SAR) analysis of pyrazole-azepinoindole derivatives.

Main Results:

• Identification of 16 selectively cytotoxic molecules, including known agents and novel compounds like macrocycle 1, pyrazole-azepinoindole derivative 2, and complex heterocyclic derivative 3.
• Confirmation of anticancer potential for seven compounds via MTT assay.
• An optimized pyrazole-azepinoindole derivative (23) exhibited a selectivity index (SI) of 18 against HCT116 versus VA-13 cells.

Conclusions:

• The FCCT is an effective method for identifying selectively cytotoxic anticancer compounds early in development.
• Novel chemical scaffolds, particularly pyrazole-azepinoindole derivatives, show significant potential for anticancer therapy.
• The identified compounds, especially derivative 23, warrant further investigation for their therapeutic efficacy, with effects mediated by G1 cell cycle arrest.