Characterization of a broad-spectrum hemp extract: analytical quantification of cannabinoids and terpenes and biological evaluation against triple-negative breast cancer cells

Triple-negative breast cancer (TNBC) remains a major therapeutic challenge due to its aggressiveness and limited treatment options. Cannabis sativa L. has gained increasing interest for its potential anticancer properties, notably through the potential of cannabinoids and terpenes1. In this study, we developed and characterized a custom-formulated hemp extract (Felina 32 variety) designed to obtain a rich phytochemical composition, with high levels of cannabidiol (CBD) and terpenes, based on our previous research on synergistic interactions to enhance the entourage effect2. This work aimed to evaluate the impact of this extract on the growth of TNBC cells and its potential impact on lymphangiogenesis.
Cannabinoid content was quantified using a validated HPLC method, while terpenes profiles were analyzed by GC, with quantitative assessment of key constituents. Antiproliferative effects were monitored kinetically over 72 hours using live-cell imaging (Incucyte SX-5, Sartorius). The extract significantly inhibited the growth of MDA-MB-231 cells, with a growth rate value of 0.18 ± 0.06 at 72 hours, indicating a near-cytostatic activity compared to control cells (GR = 1.00). Holotomographic microscopy (CX-A, Nanolive) revealed clear morphological alterations in response to treatment (loss of adherence and formation of apoptotic bodies), providing dynamic insights into cellular responses. Given the established role of lymphangiogenesis in TNBC metastasis3, additional tests were conducted on lymphatic endothelial cells (LEC) to investigate the influence of the extract on these cells. Reduction in LEC proliferation and morphological alterations were observed. Preliminary toxicity screening was performed using zebrafish embryo models to evaluate safety profiles.
These findings support the therapeutic potential of optimized hemp extracts for TNBC management and highlight the importance of preserving the complete phytochemical profile to harness potential synergistic effects.