Understanding the Antilymphoma Activity of Annona macroprophyllata Donn and Its Acyclic Terpenoids: In Vivo, In Vitro, and In Silico Studies

Abstract

Annona macroprophyllata Donn (A. macroprophyllata) is used in traditional Mexican medicine for the treatment of cancer, diabetes, inflammation, and pain. In this work, we evaluated the antitumor activity of three acyclic terpenoids obtained from A. macroprophyllata to assess their potential as antilymphoma agents. We identified the terpenoids farnesyl acetate (FA), phytol (PT) and geranylgeraniol (Gg) using gas chromatography–mass spectroscopy (GC-MS) and spectroscopic (1H, and 13C NMR) methods applied to petroleum ether extract of leaves from A. macroprophyllata (PEAm). We investigated antitumor potential in Balb/c mice inoculated with U-937 cells by assessing brine shrimp lethality (BSL), and cytotoxic activity in these cells. In addition, to assess the potential toxicity of PEAm, FA, PT and Gg in humans, we tested their acute oral toxicity in mice. Our results showed that the three terpenoids exhibited considerable antilymphoma and cytotoxic activity. In terms of lethality, we determined a median lethal dose (LD50) for thirteen isolated products of PEAm. Gg, PT and AF all exhibited a higher lethality with values of 1.41 ± 0.42, 3.03 ± 0.33 and 5.82 ± 0.58 µg mL−1, respectively. To assess cytotoxic activity against U-937 cells, we calculated the mean cytotoxic concentration (CC50) and found that FA and PT were closer in respect to the control drug methotrexate (MTX, 0.243 ± 0.007 µM). In terms of antilymphoma activity, we found that FA, PT and Gg considerably inhibited lymph node growth, with median effective doses (ED50) of 5.89 ± 0.39, 6.71 ± 0.31 and 7.22 ± 0.51 mg kg−1 in females and 5.09 ± 0.66, 5.83 ± 0.50 and 6.98 ± 0.57mg kg −1 in males, respectively. Regarding acute oral toxicity, we classified all three terpenoids as category IV, indicating a high safety margin for human administration. Finally, in a molecular docking study of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, we found binding of terpenoids to some amino acids of the catalytic site, suggesting an effect upon activity with a resulting decrease in the synthesis of intermediates involved in the prenylation of proteins involved in cancer progression. Our findings suggest that the acyclic terpenoids FA, PT, and Gg may serve as scaffolds for the development of new treatments for non-Hodgkin’s lymphoma.

In Vivo, In Vitro and In Silico Anticancer Activity of Ilama Leaves: An Edible and Medicinal Plant in Mexico

“…Several researchers have explored ilama leaves as a source of antihyperglycemiants and anticancer agents. It has been reported that ilama leaves contain flavonoid glycosides, sesquiterpenoids and an aliphatic compound [2,6].…”

“…These results suggest that the antitumoral properties, as inductors of apoptosis, of Gg and PT may be mediated by the generation of ROS. Once the induction of late apoptosis was demonstrated by the acyclic terpenoids Gg, PT, and FA, we evaluated their capability of inducing the production of ROS in the U-937 cells, considering that the production of ROS in physiologic and pathological cases may be associated with the induction of apoptosis [6]. The results showed that the terpenoids Gg and PT induced a significant increase in ROS production, with values of 58.7 and 31.5% (Table 1) of the mean fluorescence shift (Figure 3E,F).…”

“…In this context, it is important to highlight that several studies of different extracts obtained from ilama leaves have shown that they possess flavonoid glycosides, acyclic sesquiterpenoids and an aliphatic ketone. In the case of pharmacological studies, ilama leaves exhibited anxiolytic, antinociceptive, anticonvulsant, antihyperglycemic and antiepileptic effects [2][3][4][5][6].…”

“…Petroleum ether extract of leaves from Annona macroprophyllata and geranylgeraniol (Gg), phytol (PT) and farnesyl acetate (FA), three acyclic sesquiterpenoids obtained from this species of Annonaceae, exhibited antilymphoma activity in mice [6]. In this context, further studies are needed to examine the anticancer properties of A. macroprophyllata and its acyclic sesquiterpenoids using in vivo, in vitro and in silico approaches.…”

“…Cancer is a disease that affects a significant number of patients each year, and according to data from the World Health Organization, it is the leading cause of death worldwide; in 2020, it caused the death of 10 million people [7]. Around 19.3 million new cases of cancer have been diagnosed since then, and this is expected to increase in the coming years [6,8]. Non-Hodgkin's lymphoma (NHL) is the most common hematologic neoplasia characterized by the abnormal development of lymphocytes, affecting the lymph nodes, spleen, bone marrow, stomach and skin.…”