Kojic Acid from the Ethyl Acetate Fraction of Terminalia catappa Linn Fruit Flesh and its anti-cancer activity against HeLa cells

Anti-cancer activity of compounds from Bauhinia strychnifolia stem

The ethyl acetate fraction from methanol extract of ketapang (Terminalia catappa Linn) fruit flesh has anti-cancer potential. Therefore, this study aims to obtain the active anti-cervical cancer agent from the ethyl acetate fraction of ketapang fruit flesh’s methanol extract. The first step carried out was fractionation of the ethyl acetate fraction of methanol extract using gravity column chromatography with an eluent of ethyl acetate: chloroform: glacial acetic acid (1:8:1). The second step was a toxicity test on fractions produced from column chromatography. The third step was the test of secondary metabolite and anti-cancer activity of fraction possessing the highest toxicity to Artemia salina Leach. Fractionation of the concentrated ethyl acetate fraction by column chromatography, yielded seven subfractions (F1-F7). Furthermore, the cytotoxic test on A. salinaLeach shrimp larvae generated the following LC50 data from the 1st to 7th subfraction in a row, namely 566.2814 ppm, 87.9077 ppm, 216.6232 ppm, 566.2814 ppm, 560, 6647 ppm, 279.9213 ppm, and 194.6674 ppm. The most active subfraction is fraction 2 which have two groups of positive compounds, i.e. phenolics and saponins were obtained from the secondary metabolite test. Data from the infrared spectroscopy performed showed the presence of groups –OH, C-H aromatic, C=O carbonyl, C=C aromatic, C=C alkene, C-H aliphatic, C-H alkane, and C-O. The value of the anti-cancer activity of fraction 2 was IC50 = 165.37 ppm, which was included in the fairly active category as an anti-cervical cancer agent.

According to statistics, in 2012, there were estimated 1.4 million new colorectal cancer cases and 693,900 deaths. Breast cancer, the leading cause of cancer-related death among females worldwide, gave an estimated 1.7 million cases and 521,900 deaths in 2012. An estimated 527,600 cancer cases and 265,700 deaths in 2012 worldwide were caused by cervical cancer which is the third leading cause of cancer-related death in females [1]. Chemotherapeutics play an important role as anticancer agents, inducing apoptosis or restoring apoptotic functions of proteins. In view of the importance of sulfonamides and nitrogen containing heterocycles as privileged structures for designing anticancer agents, we decided to explore the synthesis and anticancer activity of molecular hybrids obtained by the combination of benzenesulfonamide and heterocycles such as imidazole, 1,2,4-triazole, benzimidazole and benzoxazole. The anticancer activities of compounds were evaluated in vitro on MCF-7, HCT-116 and HeLa human tumor cell lines by MTT assay. The most active compounds bearing 3-methyl-2-thioxo-1H-imidazol-1-yl moiety exhibited selectivity against HeLa cells with IC50 values 6‒7 µM. Meanwhile, 2-thioxo-1H-benzo[d]imidazole derivatives showed activity against HCT-116 cells in the range of IC50: 17‒36 µM. The apoptotic potential of the most active compounds was analyzed through various assays in HeLa cells: phosphatidylserine translocation, cell cycle dystribution and caspase activation. Results indicated that compounds promoted cell cycle arrest at sub-G1 phase in cancer cells, induced caspase activity and increased the population of apoptotic cells. [1] Torre L.A. et. al. Cancer Epidemiol. Biomarkers Prev. 25 (2016) 16‒27.

Secang (Caesalpinia sappan L.) contains potentially anti-cancer active compounds, namely brazilin and brazilein which are soluble in ethyl acetate. This study is conducted to evaluate the anticancer activity of secang wood in cervical cancer cells (HeLa) in vitro and molecular mechanisms that selectively mediate their cytotoxic effects on several cancer cells through molecular docking or in silico. The anticancer potential of the ethyl acetate fraction of secang wood through its cytotoxic activity on HeLa cells was carried out with MTT assay and calculated the IC50 value. The molecular mechanism is carried out in silico with the target protein p53 and caspase 9 using the autodock 4.2 program. The docking process includes protein and active compounds preparation, validation of molecular docking methods and docking brazilein as the active substance with the target protein. The docking score is measured by binding energy value between brazilein and the target protein. The fraction of secang wood was cytotoxic in HeLa cells with IC50 values of 48.71 μg/mL. The docking results showed that brazilein had an affinity for the target proteins p53 and caspase 9 with binding energies of -8.24 and -6.71 kcal / mol, respectively.

Eleutherine palmifolia (L.) Merr. is a typical plant found in Central Kalimantan that has been used empirically by the Dayak people as medicine for various diseases, including cancer. The plant contains flavonoid compounds that potentially used as an anticancer. The purpose of this study is to find the most active fraction, indicated by its cytotoxic potency on HeLa cervical cancer cell line, and to identify compounds in E. palmifolia bulbs fraction. E. palmifolia bulbs was extracted by maceration. The extraction with ultrasonic bath and partition fractionation was conducted by using n-hexane, chloroform, and ethyl acetate. Each fraction was tested for toxicity level on HeLa cells using MTT assay. The identification of active compounds was carried out by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS). The result showed that based on the IC50 value, the ethyl acetate fraction had the highest bioactivity. IC50 values of n-hexane, chloroform, and ethyl acetate fractions were 250.77±19.01; 720.46±42.38; and 44.34±9.45μg/mL, respectively. The identification of the active compound in ethyl acetate fraction resulted 28 chemical compounds. Compounds with the highest percentage area were isoliquiritigenin and oxyresveratrol. The ethyl acetate fraction of E. palmifolia bulbs is potential to be developed as an anticancer candidate (phytopharmaceutical).Keywords: Compound identification, Anticancer activity, Eleutherine palmifolia (L.) Merr., cervical cancer

BackgroundEichhornia crassipes (Mart) solms is an invasive macrophyte causing serious problems to the network of irrigation and drainage canals in the Nile Delta region. The present study aim to evaluate the potential anticancer and antioxidant activities of Eichhornia crassipes crude extract and its pure compounds.MethodsThe macrophyte was collected from El-Zomor canal, River Nile (Egypt), cleaned, air dried, grinded then extracted with methanol (crude extract). The extract was fractionated using pre-coated silica gel plates (TLC F254) with hexane/ethyl acetate (8.5: 1.5 v/v) as mobile phase. Nine fractions were separated (A-I) then scratched, eluted with the same mobile phase, filtered and the separated fractions were determined and identified using spectroscopic methods (Mass spectrum (MS), Infra red (IR) and Proton H-Nuclear magnetic resonance (H-NMR). Both the crude extract and its nine identified compounds were tested for their antioxidant (using 2, 2 diphenyl-1-picrylhydrazyl (DPPH), 2, 2′- azino-bis {ethylbenzthiazoline-6-sulfonic acid (ABTS.)} methods) and anticancer activity (using MCF-7, HeLa, Hep.G2 and EACC cell lines).ResultsThe antioxidant and anticancer activities of the crude extract exhibited the highest effect while the compounds showed variable effects which depend on the type of compound and cancer cell line. The antioxidant activity of the crude extract exhibited the highest followed in descending order by compounds D, E, G and H respectively. Concerning the anticancer potency, the crude extract showed also the highest effect while the identified compounds (A, B, C, D, E, F, G, H and I) showed variable anticancer activities against the four different cell lines. In addition, Compound I exhibited the most potent anticancer activity against HepG2 cell line while compound D exhibited high anticancer activity against HeLa cells and EACC. The results revealed the presence of different compounds (Alkaloids and terpenoids) with variable antioxidant and anticancer activities which elicited an auto-augmentation in the crude extract leading to its greatest activities. The action of the identified anticancer compounds on DNA fragmentation was studied.ConclusionThe study illustrated the potential of Eichhornia as a valuable resource for natural compounds of desirable medicinal properties (e.g. antioxidants and anticancer).

Erythrina caffra is a traditional plant used to treat cancer and inflammation. The study aimed to assess and isolate anticancer compounds from E. caffra bark. The plant material was extracted sequentially in n-hexane, dichloromethane, ethyl acetate and methanol. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and 3-(4,5-di methyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were used to evaluate the crude extracts’ antioxidant and anticancer activities, respectively. Column chromatography was used to purify the potent extracts of the stem bark in order to isolate the bioactive compounds. The crude extracts of the E. caffra bark demonstrated antioxidant and anticancer activity, with the dichloromethane (DCM) extract producing the most favorable activity. Three compounds, namely Hexacosanyl isoferulate, Tetradecyl isoferulate, and 1-Heneicosanol, were detected in fractions from the DCM extract. All the isolated compounds showed significant anticancer potential, with the hydroxycinnamic acid compounds showing better anticancer effects in the cervical (HeLa) and breast cancer (MCF-7) cells. The compounds showed greater activity than even the standard drug, 5-fluorouracil, in the MCF-7 cells, with the tetradecyl isoferulate and hexacosanyl isoferulate fractions having IC50 values of 123.62 and 58.84 µg/mL, respectively. The compounds were observed to be capable of triggering caspase cascade events, leading to apoptotic cell death. Overall, E. caffra extracts contained important bioactive compounds that induced apoptotic cell death in HeLa and MCF-7 tumor cells, warranting further investigations in vitro and in vivo.

Sea Cucumber Holothuria atra is one of marine organisms has been used as a new source of novel bioactive compounds. Many of them have been used as the lead compounds in discovery of new anticancer drugs. The objective of this study was to determine the active fractions of sea cucumber (H. atra) which have anticancer activity. H. atra was macerated using ethanol and the extract was freezedried using a freeze dryer. The crude extract was partitioned using n-hexane, ethyl acetate, and methanol-water (3:1:1:1). Cytotoxicity test was performed using HeLa (cervic cancer) cell line and MCF-7 (breast cancer) cell line based on the MTT assay. The crude extract of H. atra showed the best cytotoxic activity against HeLa cells (IC50 = 12.48 µg/mL) and MCF-7 cells (IC50 = 17.90 µg/mL). The toxicity tests showed the IC50 value of the n-hexane fraction, ethyl acetate fraction, and methanol-water fraction against HeLa cells HeLa (IC50 = 76.45 µg/mL; 77.95 µg/mL; 14.27 µg/mL) and MCF-7 cells (IC50 = 58.50 µg/mL; 59.59 µg/mL; 14.33 µg/mL).

Fucoidan (F) is an anticancer potential natural compound extracted from the brown seaweed Sargassum plagiophyllum. However, poor solubility and lack of targeted delivery are major drawbacks in its therapeutic application. The aim of this study is to enhance the delivery and efficacy of fucoidan by encapsulating it using -carrageenan (C) and Carrageenan folate (Cf) as carrier matrices. The particle size of the resulting fucoidan-loaded nanocapsules with -Carrageenan (F-CNPs) and -Carrageenan folate (F-CfNPs), were 84.212.1nm and 93.210.7nm, respectively. Encapsulation Efficiency (EE) was high, with 92.340.58% for F-CNPs and 95.41 0.06% for F-CfNPs, while the Loading amount (LA) were 46.170.29% and 47.700.03%, respectively. Anticancer activities of the nanocapsules against HeLa and MCF-7 cell lines were performed. The IC50 values in F-CfNPs were significantly lower compared with F-CNPs, representing increased efficacy due to the addition of the targeting folate group. In HeLa cells, the IC50 values were 33.134.53g ml-1 for F-CNPs, 21.143.59 g ml-1 for F-CfNPs, and 3.163.56 g ml-1 for doxorubicin. While, in MCF-7 cells, the IC50 values were 30,563.86 g ml-1 for F-CNPs, 24.923.83g ml-1 for F-CfNPs, and 15.792.84g ml-1 for doxorubicin. These results clearly indicated that the fucoidan nanocapsules acted as a potent therapeutic against HeLa and MCF.7 cell lines. However, F-CfNPs showed higher efficiency among the fabricated NPs due to higher cellular uptake. This study toward the 14th Sustainable Development Goals (SDG) by utilizing brown seaweed to enhance its economic value and aligns with the 3rd SDG goal of ensuring good health well-being through improvements in cancer treatment.

Anticancer activity in HeLa and MCF-7 cells via apoptopic cell death by a sterol molecule Cholesta-4,6-dien-3-ol (EK-7), from the marine ascidian Eudistoma kaverium

In the present study, the synthesis of benzimdazolyl 2-amino-1,3,4-oxadiazole derivatives (5a-j) and their in vitro anticancer activities against HeLa, MCF7, A549, and HEK293 cell lines were reported. Compound 5f showed most promising anticancer activity with IC50 values 4.68 ± 0.04, 4.16 ± 0.02, 5.40 ± 0.08 µM against the HeLa, MCF-7, and A549 cell lines respectively. Compounds 5b and 5e have also shown excellent anticancer activity with IC50 values 6.07 ± 0.028, 5.30 ± 0.09, 7.16 ± 0.061 µM and 7.56 ± 0.073, 7.20 ± 0.048, 11.30 ± 0.018 µM against the HeLa, MCF-7, and A549 cell lines respectively. The rest of the compounds displayed moderate to good anticancer activity. The tested compounds were nontoxic to normal HEK293 cell lines, and even the more active compounds (5b, 5e, and 5f) also less toxic on HEK-293 cells. Molecular docking results of the synthesized compounds with the target EGFR protein were also discussed.

The leaves of Garcinia celebica strongly inhibit the proliferation of MCF-7 human breast adenocarcinoma cell lines. The present study focused on investigating the active anticancer and antiproliferative compound from the G. celebica leaves and assessing its mechanism of action. Ethanol extracts of G. celebica were fractionated based on their polarity using n-hexane, ethyl acetate and water. The antiproliferative properties were tested in vitro against MCF-7 human breast cancer cell lines using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. The active compound was subsequently isolated using column chromatography and identified by nuclear magnetic resonance. The characterized compound was also tested for its antiproliferative properties and the mechanism by which it induces apoptosis in MCF-7 cells by western blot analysis of the activated apoptotic proteins. This resulted in the isolation of a friedolanostane triterpenoid, which was determined to be methyl-3α, 23-dihydroxy-17,14-friedolanstan-8,14,24-trien-26-oat. This compound inhibited MCF-7 cell proliferation in a time- and dose-dependent manner with IC50 values of 82 and 70 µM for the 24 and 48 h treatments, respectively. Furthermore, the western blot analysis suggested that the compound exerted its anticancer activities by promoting apoptosis through the inhibition of the oncogenic protein Akt, thereby increasing the expression of poly (ADP-ribose) polymerase (PARP) protein. These results suggest that methyl-3α,23-dihydroxy-17,14-friedolanstan-8,14,24-trien-26-oat is the anticancer compound found in G. celebica, providing a basis for its potential use in cancer disease management.

A study was conducted to determine the antioxidant, antibacterial, cytotoxic, and anticancer activities of the belulang grass plant (Eleusine indica). Antioxidant activity was assessed using the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity method. Antibacterial activity was determined using the disk diffusion method, cytotoxicity was determined using the Brine Shrimp Lethality Test (BSLT), and anticancer activity was evaluated using the MTS assay. The results showed that the antioxidant activity of the extracts and solvent fractions of Eleusine indica had the highest average value for the n-hexane fraction (FNH) at 35.0%, ethyl acetate fraction (FEA) at 56.4%, aqueous fraction (FA) at 30.3%, and methanol extract (EM) at 32.5%. Among these, the highest value was obtained in FEA, while the lowest was obtained in FA. Based on the cytotoxicity test with the BSLT method, the ethyl acetate fraction had an LC50 value of 7.384 ppm, categorizing it as highly toxic. However, in cytotoxicity testing with HeLa cells, the IC50 value for the ethyl acetate fraction was 223.70 ppm, indicating that the ethyl acetate fraction was less active against HeLa cervical cancer cells. Keywords: Eleusine indica; belulang grass; cytotoxic; HeLa cells

Kojic acid has gained its importance after it was known worldwide that the substance functions primarily as skin-lightening agent. Kojic acid plays a vital role in skin care products, as it enhances the ability to prevent exposure to UV radiation. It inhibits the tyrosinase formation which suppresses hyperpigmentation in human skin. Besides cosmetics, kojic acid is also greatly used in food, agriculture, and pharmaceuticals industries. Conversely, according to Global Industry Analysts, the Middle East, Asia, and in Africa especially, the demand of whitening cream is very high, and probably the market will reach to $31.2 billion by 2024 from $17.9 billion of 2017. The important kojic acid-producing strains were mainly belongs to the genus Aspergillus and Penicillium. Due to its commercial potential, it continues to attract the attention for its green synthesis, and the studies are still widely conducted to improve kojic acid production. Thus, the present review is focused on the current production processes, gene regulation, and limitation of its commercial production, probable reasons, and possible solutions. For the first time, detailed information on the metabolic pathway and the genes involved in kojic acid production, along with illustrations of genes, are highlighted in the present review. Demand and market applications of kojic acid and its regulatory approvals for its safer use are also discussed. KEY POINTS: • Kojic acid is an organic acid that is primarily produced by Aspergillus species. • It is mainlyusedin the field of health care and cosmetic industries. • Kojic acid and its derivatives seem to be safe molecules for human use.

Calotropis gigantea (L.) W. Aiton (C. gigantea) is a medicinal plant that has been empirically proven to have anticancer activity. In a previous study, it showed that the fraction of ethyl acetate from the root part of C. gigantea had higher anticancer activity than the other fractions. It suspected that the ethyl acetate fraction of C. gigantea root contained active compounds that has anticancer properties. This study aimed to determine the anticancer activity of active compounds from the ethyl acetate fraction of C. gigantea root regarding induction of apoptosis, cell cycle arrest, and expression of caspase-8 colon cancer cell WiDr. Isolation of the active compounds from the ethyl acetate fraction of C. gigantea root was carried out using Bioassay-guided Isolation method. Identification of active compounds was using NMR-1H, NMR-13C, HMBC, HMQC and UPLCMS/MS methods. The anticancer activity test of the identified compounds performed by using MTT method. The induction of apoptotic and cell cycle arrest evaluated by a flow cytometry method. The result of this study showed two active compounds were identified namely (1) (Pregnanon-5-en, 3,14,17 trihydroxy-12- (4’-cyclohexyl benzoate) -, (3β, 12β, 14β) - (9CI), (2) Pregn-5-en-20-one, 3,8,14-trihydroxy-12 - [(4’-hydroxy benzoyl) oxy] -, (3β, 12β, 14β, 17α) - (9CI). Both compounds inhibited the growth of colon cancer cell WiDr with IC50 values respectively were 15.89 μg/mL and 0.77 μg/mL. Both compounds increased the induction of apoptotic by increasing sub-G1, S, and G2-M following depletion of G0-G1 phase accumulation.