Betulin is a birch bark-derived lupane-type pentacyclic triterpene with a wide spectrum of biological activities. Given their enhanced antiproliferative potential and enhanced pharmacological profile, betulin derivatives are continuously investigated in scientific studies. The objective of the current study was to in vitro assess the antiproliferative properties of novel synthesized 1,2,4-triazole derivatives of diacetyl betulin. The compounds were investigated using three cancer cell lines: A375 (melanoma), MCF-7 (breast cancer), HT-29 (colorectal cancer), and HaCaT (human keratinocytes). Bet-TZ1 had the lowest recorded IC₅₀ values (ranging from 22.41 to 46.92 μM after 48 h of exposure) than its precursor and other tested compounds in every scenario, with the highest cytotoxicity against the A375 cell line. Bet-TZ3 demonstrated comparable cytotoxicity to the previously mentioned compound, with an IC₅₀ of 34.34 μM against A375. Both compounds caused apoptosis in tested cells, by inducing specific nuclear morphological changes and by increasing the expression of caspase 9, indicating significant cytotoxicity, which was consistent with the literature and viability evaluation. Bet-TZ1 and Bet-TZ3 inhibit cancer cell migration, with the former having a stronger effect than the latter. The HET−CAM test indicated that all compounds have no irritative potential, suggesting that they can be used locally.

Betulinic acid is a lupane-type pentacyclic triterpene mostly found in birch bark and thoroughly explored for its wide range of pharmacological activities. Despite its impressive biological potential, its low bioavailability has challenged many researchers to develop different formulations for achieving better in vitro and in vivo effects. We previously reported the synthesis of fatty acid esters of betulinic acid using butyric, stearic, and palmitic acids (But-BA, St-BA, and Pal-BA) and included them in surfaced-modified liposomes (But-BA-Lip, St-BA-Lip, Pal-BA-Lip). In the current study, we evaluated the cytotoxic effects of both fatty acid esters and their respective liposomal formulations against MCF-7, HT-29, and NCI-H460 cell line. The cytotoxic assessment of BA derivatives revealed that both the fatty esters and their liposomal formulations acted as cytotoxic agents in a dose- and time-dependent manner. But-BA-Lip exerted stronger cytotoxic effects than the parent compound, BA and its liposomal formulation, and even stronger effects than 5-FU against HT-29 cells (IC₅₀ of 30.57 μM) and NCI-H460 cells (IC₅₀ of 30.74 μM). BA’s fatty esters and their respective liposomal formulations facilitated apoptosis in cancer cells by inducing nuclear morphological changes and increasing caspase-3/-7 activity. The HET-CAM assay proved that none of the tested compounds induced any irritative effect, suggesting that they can be used safely for local applications.

Betulinic acid is a naturally occurring pentacyclic triterpene belonging to the lupane-group that exhibits a wide range of pharmacological activities. BA derivatives are continuously being researched due to their improved anticancer efficacy and bioavailability. The current research was conducted in order to determine the antiproliferative potential of three synthesized BA fatty esters using palmitic, stearic and butyric acids and their liposomal nanoformulations. The cytotoxic potential of BA fatty esters (Pal-BA, St-BA, But-BA) and their respective liposomal formulations (Pal-BA-Lip, St-BA-Lip, But-BA-Lip) has been assessed on HaCaT immortalized human keratinocytes and A375 human melanoma cells. Both the esters and their liposomes acted as cytotoxic agents against melanoma cells in a time- and dose-dependent manner. The butyryl ester But-BA outperformed BA in terms of cytotoxicity (IC₅₀ 60.77 μM) while the nanoformulations St-BA-Lip, But-BA-Lip and BA-Lip also displayed IC₅₀ values (60.11, 50.71 and 59.01 μM) lower compared to BA (IC₅₀ 65.9 μM). The morphological evaluation revealed that the A375 cells underwent morphological changes consistent with apoptosis following 48 h treatment with the tested compounds, while the HaCaT cells’ morphology remained unaltered. Both the esters and their liposomal formulations were able to inhibit the migration of the melanoma cells, suggesting a significant antimetastatic effect. The quantitative real-time PCR revealed that all tested samples were able to significantly increase the expression of the pro-apoptotic Bax and inhibit the anti-apoptotic Bcl-2 proteins. This effect was more potent in the case of liposomal nanoformulations versus non-encapsulated compounds, and overall, But-BA and its formulation exhibited the best results in this regard.

Oregano essential oil (OEO) has gained attention for its broad pharmacological activities, such as anti-inflammatory, antimicrobial, and anticancer properties. This study aimed to analyze the phytochemical composition and biological activity of OEO obtained from wild-growing Origanum vulgare L. in Romania. Gas chromatography–mass spectrometry (GC–MS) analysis identified p-cymene (43.98%), γ-terpinene (22.16%), and thymol (11.46%) as major constituents, with notable differences from previously reported chemotypes. Antioxidant activity was assessed using the DPPH, ABTS radical scavenging assay, and TPC. OEO has a moderate antioxidant activity, with IC₅₀ values of 134.67 ± 1.32 µg/mL (DPPH) and 88.15 ± 0.045 Inh% (ABTS) and a TPC of 159.63 mg GAE/g extract. The cytotoxicity of the simple water dispersion of OEO, OEO solubilized with polyethylene glycol 400 (OEO-PEG), and that solubilized with Tween 20 (OEO-Tw) was evaluated on human melanoma (A375) and human colorectal adenocarcinoma (HT-29) cancer cell lines, as well as on the normal human immortalized keratinocytes (HaCaT) cell line. The results demonstrated a significant inhibition of cancer cell viability with no recorded cytotoxic effect on normal cells. The highest inhibition of cell viability was recorded for OEO-PEG 200 µg/mL (7.22% ± 6.51 in A375 cell line and 22.25% ± 10.08 in HT-29 cell line). In cancer cells, OEO and its formulations significantly reduced malondialdehyde (MDA) levels (up to 41.24% in A375 cells and up to 48.58% in HT-29 cells), suggesting potent antioxidant activity. Moreover, treatment with OEO increased caspase 3/7 activation two-fold in treated A375 cells, while high-resolution respirometry studies revealed that OEO induces mitochondrial dysfunction by acting as a potential uncoupling agent. Molecular docking analysis suggested that β-caryophyllene oxide (CPO), a minor constituent of OEO, may act as a potential inhibitor of 3-phosphoinositide-dependent protein kinase-1 (PDPK1), indicating a possible mechanism of anticancer activity. Our findings highlight the potential of OEO as a natural anticancer agent, emphasizing the need for further investigations to elucidate its exact molecular mechanisms and therapeutic applicability.

Probiotic strains such as Lactobacillus spp. are already known for their beneficial effect on human health and new research supports their role in colon cancer prevention and treatment. The current study reports the effect of different concentrations of Lacticaseibacillus rhamnosus (LGG, 10⁶–10⁹ CFU/mL), alone or in association with 5-fluorouracil (5-FU, 10 μM), tested against normal HaCaT cells, HT-29 colorectal adenocarcinoma and HCT-116 colorectal carcinoma cell lines. The underlying cytotoxic effect was further investigated. LGG treatment of HT-29 and HCT-116 cells caused a variety of apoptotic-related nuclear morphological changes, as revealed by DAPI staining. ELISA studies showed that LGG treatment increased caspase-3 activity and pro-apoptotic BAX protein levels while decreasing anti-apoptotic Bcl-2 protein levels and the proto-oncogene Cyclin D1. A more detailed examination of the mitochondrial function revealed that high concentrations of LGG can impair mitochondrial function in HT-29 and HCT-116 cancer cells. All of these findings suggest that LGG has a pro-apoptotic, mitochondrial-targeted, cytotoxic effect on both colon cancer cell lines studied.

Background/Objectives: One of the major problems facing the medical world is cancer, which causes millions of deaths each year. Nitrogen-containing heterocycles appear to offer a potential solution to this problem. Thus, the present study aimed to synthesize and characterize—both physicochemically and biologically—the new compounds obtained by combining benzotriazole and 1,2,4-triazole scaffolds. Methods: Each of the four cell lines (HaCaT—human keratinocyte, A375—melanoma, A549—non-small cell lung cancer, and HT-29—colorectal adenocarcinoma) was treated for 48 h with concentrations ranging from 10 to 100 µM of the four compounds. After 48 h of treatment, changes in cell viability were assessed using the Alamar Blue assay. These findings were supplemented by microscopic analysis of metabolic changes, as well as alterations in the actin cytoskeleton and nuclei, using staining techniques. Results: The results showed that MG-1 (p-chlorophenyl-) and MG-3 (p-(N,N-dimethylamino)phenyl-) are the most active compounds in our study, with IC₅₀ values for each tumor line as follows: 75.43 ± 2.77, 77.15 ± 1.32, and 96.28 ± 1.72 μM in A375, A549, and HT-29 cells, and 78.97 ± 3.08, 56.78 ± 2.57, and 69.19 ± 3.97 μM, respectively. In HaCaT cells, IC₅₀ values remained above 100 μM for all four hybrids. In silico screening identified MEK1 as the most favorable target for the investigated series, while 100 ns molecular dynamics simulations and MM-GBSA analysis supported stable binding of MG-1 and MG-3, with predicted binding free energies of −54.83 and −40.70 kcal/mol, respectively. Conclusions: Even with a limited set of compounds, we can conclude that the biological effect observed in our study depended on both the cell line and the substituent attached to the aryl moiety. The results obtained for MG-1 and MG-3 suggest that these compounds may serve as suitable starting points for further structural optimization and investigation of the mechanisms of action.