Poor prognosis of most cancer patients is in part, due to limited therapeutic options. Furthermore, as chemotherapy remains the standard-of-care for several cancers, partial or lack of response remains a concern and compounding this are the adverse side effects of the treatment that severely impacts the quality of life and survival. In pursuit of improving treatment options, we have opted to investigate the unique chemical skeleton of natural compounds as anticancer therapies. In this study, from an initial screen of 31 crude methanol extracts from ~15 plant species using HL60 cells, the root extract of Brucea javanica (L.) Merr indicated the presence of bioactive compounds. Subsequent bioassay-guided purification on the root extract yielded two alkaloids canthin-6-one (1) and bruceolline J (2), which were further investigated for their bioactivity in representative human cancer lines and normal phenotypic counterparts. MTT assay demonstrated ED50 values from 34.7–72.9 μM for 1 and 16.0–54.0 μM for 2 for the cancer cell lines panel. NP69 cells also demonstrated sensitivity to both compounds (9.3 μM and 4.5 μM). As amount of 2 isolated were limiting, we focused on 1 to further identify novel anticancer properties in PC3 and HeLa cancer lines. We observed at 30 μM, 1 induced a G2/M phase arrest coinciding with decreased cell proliferation. Furthermore, 1 was able to synergize the cytotoxic effect of cisplatin when used in combination, suggesting the potential of combination therapy for those less responsive lesions to standard chemotherapy.
FerlayJ, SoerjomataramI, DikshitR, EserS, MathersC, RebeloM, ParkinDM, FormanD, BrayF. (2015) Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. International Journal of Cancer, 136, E359–386.
2.
OmarZ, Ibrahim TaminN. (2011) National cancer registry report: Malaysia cancer statistics-data and figure. Malaysia: National Cancer Registry, 85–87.
3.
FennellDA, SummersY, CadranelJ, BenepalT, ChristophDC, LalR, DasM, MaxwellF, Visseren-GrulC, FerryD. (2016) Cisplatin in the modern era: The backbone of first-line chemotherapy for non-small cell lung cancer. Cancer Treatment Reviews, 44, 42–50.
4.
MarkmanM. (2013) Current standards of care for chemotherapy of optimally cytoreduced advanced epithelial ovarian cancer. Gynecologic Oncology, 131, 241–245.
5.
PlimackER, DunbrackRL, BrennanTA, AndrakeMD, ZhouY, SerebriiskiiIG, SlifkerM, AlpaughK, DulaimiE, PalmaN, Hoffman-CensitsJ, BilusicM, WongYN, KutikovA, ViterboR, GreenbergRE, ChenDY, LallasCD, TrabulsiEJ, YelenskyR, McConkeyDJ, MillerVA, GolemisEA, RossEA. (2015) Defects in DNA repair genes predict response to neoadjuvant cisplatin-based chemotherapy in muscle-invasive bladder cancer. European Urology, 68, 959–967.
6.
ChangA. (2011) Chemotherapy, chemoresistance and the changing treatment landscape for NSCLC. Lung cancer, 71, 3–10.
7.
EltingLS, CooksleyC, ChambersM, CantorSB, ManzulloE, RubensteinEB. (2003) The burdens of cancer therapy. Cancer, 98, 1531–1539.
8.
ChienM, AstumianM, LiebowitzD, Rinker-SchaefferC, StadlerWM. (1999) In vitro evaluation of flavopiridol, a novel cell cycle inhibitor, in bladder cancer. Cancer Chemotherapy and Pharmacology, 44, 81–87.
9.
YasuiH, ImuraY, OutaniH, HamadaKI, NakaiT, YamadaS, TakenakaS, SasagawaS, ArakiN, ItohK, MyouiA, YoshikawaH, NakaN. (2016) Trabectedin is a promising antitumour agent for synovial sarcoma. Journal of Chemotherapy, 1–8.
10.
ShahMA, KortmanskyJ, MotwaniM, DrobnjakM, GonenM, YiS, WeyerbacherA, Cordon-CardoC, LefkowitzR, BrennerB, O'ReillyE, SaltzL, TongW, KelsenDP, SchwartzGK. (2005) A phase I clinical trial of the sequential combination of irinotecan followed by flavopiridol. Clinical Cancer Research, 11, 3836–3845.
11.
MishraBB, TiwariVK. (2011) Natural products: an evolving role in future drug discovery. European Journal of Medicinal Chemistry, 46, 4769–4807.
12.
NewmanDJ, CraggGM. (2012) Natural products as sources of new drugs over the 30 years from 1981 to 2010. Journal of Natural Products, 75, 311–335.
13.
El-ElimatT, ZhangX, JarjouraD, MoyFJ, OrjalaJ, KinghornAD, PearceCJ, OberliesNH. (2012) Chemical diversity of metabolites from fungi, cyanobacteria, and plants relative to FDA-approved anticancer agents. ACS Medicinal Chemistry Letters, 3, 645–649.
14.
BaillyC. (2009) Ready for a comeback of natural products in oncology. Biochemical Pharmacology, 77, 1447–1457.
15.
ZhangG, LiJ, ZhuT, GuQ, LiD. (2016) Advanced tools in marine natural drug discovery. Current Opinion in Biotechnology, 42, 13–23.
16.
Megadiverse Countries, United Nations Environment Programme World Conservation Monitoring Centre, accessed on 10 January 2016, http://www.biodiversitya-z.org.
17.
KimIH, SuzukiR, HitotsuyanagiY, TakeyaK. (2003) Three novel quassinoids, javanicolides A and B, and javanicoside A, from seeds of Brucea javanica. Tetrahedron, 59, 9985–9989.
18.
KimIH, TakashimaS, HitotsuyanagiY, HasudaT, TakeyaK. (2004) New quassinoids, javanicolides C and D and javanicosides B-F, from seeds of Brucea javanica. Journal of Natural Products, 67, 863–868.
19.
LiuJH, QinJJ, JinHZ, HuXJ, ChenM, ShenYH, YanSK, ZhangWD. (2009) A new triterpenoid from Brucea javanica. Archives of Pharmacal Research, 32, 661–666.
20.
PanL, ChinYW, ChaiHB, NinhTN, SoejartoDD, KinghornAD. (2009) Bioactivity-guided isolation of cytotoxic constituents of Brucea javanica collected in Vietnam. Bioorganic & Medicinal Chemistry, 17, 2219–2224.
21.
ChenM, ChenR, WangS, TanW, HuY, PengX, WangY. (2013) Chemical components, pharmacological properties, and nanoparticulate delivery systems of Brucea javanica. International Journal of Nanomedicine, 8, 85–92.
22.
GrosvenorPW, GothardPK, McWilliamNC, SuprionoA, GrayDO. (1995) Medicinal plants from Riau province, Sumatra, Indonesia. Part 1: Uses. Journal of Ethnopharmacology, 45, 75–95.
23.
SawangjaroenN, SawangjaroenK. (2005) The effects of extracts from anti-diarrheic Thai medicinal plants on the in vitro growth of the intestinal protozoa parasite: Blastocystis hominis. Journal of Ethnopharmacology, 98, 67–72.
24.
HoutS, CheaA, BunSS, EliasR, GasquetM, Timon-DavidP, BalansardG, AzasN. (2006) Screening of selected indigenous plants of Cambodia for antiplasmodial activity. Journal of Ethnopharmacology, 107, 12–18.
25.
NoorShahidaA, WongTW, ChooCY. (2009) Hypoglycemic effect of quassinoids from Brucea javanica (L.) Merr (Simaroubaceae) seeds. Journal of Ethnopharmacology, 124, 586–591.
26.
YangJ, LiS, XieC, YeH, TangH, ChenL, PengA. (2013) Anti-inflammatory activity of ethyl acetate fraction of the seeds of Brucea javanica. Journal of Ethnopharmacology, 147, 442–446.
27.
BawmS, MatsuuraH, ElkhateebA, NabetaK, Subeki NonakaN, OkuY, KatakuraK. (2008) In vitro antitrypanosomal activities of quassinoid compounds from the fruits of a medicinal plant, Brucea javanica. Veterinary Parasitology, 158, 288–294.
28.
ShenJG, ZhangZK, WuZJ, OuyangMA, XieLH, LinQY. (2008) Antiphytoviral activity of bruceine-D from Brucea javanica seeds. Pest Management Science, 64, 191–196.
29.
AlvesIABS, MirandaHM, SoaresLAL, RandauKP. (2014) Simaroubaceae family: botany, chemical composition and biological activities. Revista Brasileira de Farmacognosia, 24, 481–501.
30.
LauST, LinZX, LiaoY, ZhaoM, ChengCH, LeungPS. (2009) Bruceine D induces apoptosis in pancreatic adenocarcinoma cell line PANC-1 through the activation of p38-mitogen activated protein kinase. Cancer Letters, 281, 42–52.
31.
OhnishiS, FukamiyaN, OkanoM, TagaharaK, LeeKH. (1995) Bruceosides D, E, and F, three new cytotoxic quassinoid glucosides from Brucea javanica. Journal of Natural Products, 58, 1032–1038.
32.
VieiraIJC, Braz-FilhoR. (2006) Quassinoids: structural diversity, biological activity and synthetic studies. Studies in Natural Products Chemistry, 33, 433–492.
33.
FiaschettiG, GrotzerMA, ShalabyT, CastellettiD, ArcaroA. (2011) Quassinoids: from traditional drugs to new cancer therapeutics. Current Medicinal Chemistry, 18, 316–328.
34.
TranTV, MalainerC, SchwaigerS, AtanasovAG, HeissEH, DirschVM, StuppnerH. (2014) NF-kappaB inhibitors from Eurycoma longifolia. Journal of Natural Products, 77, 483–488.
35.
OngCY, LingSK, AliRM, CheeCF, SamahZA, HoAS, TeoSH, LeeHB. (2009) Systematic analysis of in vitro photo-cytotoxic activity in extracts from terrestrial plants in Peninsula Malaysia for photodynamic therapy. Journal of Photochemistry and Photobiology. B, Biology, 96, 216–222.
36.
WuJ, ZhouT, ZhangSW, ZhangXH, XuanLJ. (2009) Cytotoxic terpenoids from the fruits of Vitex trifolia L. Planta Medica, 75, 367–370.
37.
TanPJ, OngCY, DanialA, YusofHM, NeohBK, LeeHB. (2011) Cyclic tetrapyrrolic photosensitisers from the leaves of Phaeanthus ophthalmicus. Chemistry Central Journal, 5, 32.
38.
AndersonLA, HarrisA, PhillipsonJD. (1983) Production of cytotoxic canthin-6-one alkaloids by Ailanthus altissima plant cell cultures. Journal of Natural Products, 46, 374–378.
39.
KanchanapoomT, KasaiR, ChumsriP, HiragaY, YamasakiK. (2001) Canthin-6-one and beta-carboline alkaloids from Eurycoma harmandiana. Phytochemistry, 56, 383–386.
40.
LiuKC, YangSL, RobertsMF, PhillipsonJD. (1990) Production of canthin-6-one alkaloids by cell suspension cultures of Brucea javanica (L.) Merr. Plant Cell Reports, 9, 261–263.
41.
ChenH, BaiJ, FangZF, YuSS, MaSG, XuS, LiY, QuJ, RenJH, LiL, SiYK, ChenXG. (2011) Indole alkaloids and quassinoids from the stems of Brucea mollis. Journal of Natural Products, 74, 2438–2445.
42.
KuoPC, ShiLS, DamuAG, SuCR, HuangCH, KeCH, WuJB, LinAJ, BastowKF, LeeKH, WuTS. (2003) Cytotoxic and antimalarial beta-carboline alkaloids from the roots of Eurycoma longifolia. Journal of Natural Products, 66, 1324–1327.
43.
LiuJH, JinHZ, ZhangWD, YanSK, ShenYH. (2009) Chemical constituents of plants from the genus Brucea. Chemistry & Biodiversity, 6, 57–70.
44.
MalaysiaMoH. (2007) National Cancer Registry Report 2007.
45.
HanahanD, WeinbergRA. (2011) Hallmarks of cancer: the next generation. Cell, 144, 646–674.
46.
DejosC, VoisinP, BernardM, RegnacqM, BergesT. (2014) Canthin-6-one displays antiproliferative activity and causes accumulation of cancer cells in the G2/M phase. Journal of Natural Products, 77, 2481–2487.
47.
ClarkAS, KarasicTB, DeMicheleA, VaughnDJ, O'HaraM, PeriniR, ZhangP, LalP, FeldmanM, GallagherM, O'DwyerPJ. (2016) Palbociclib (PD0332991)-a selective and potent cyclin-dependent kinase inhibitor: a review of pharmacodynamics and clinical development. JAMA Oncology, 2, 253–260.
48.
VoraSR, JuricD, KimN, Mino-KenudsonM, HuynhT, CostaC, LockermanEL, PollackSF, LiuM, LiX, LeharJ, WiesmannM, WartmannM, ChenY, CaoZA, Pinzon-OrtizM, KimS, SchlegelR, HuangA, EngelmanJA. (2014) CDK 4/6 inhibitors sensitize PIK3CA mutant breast cancer to PI3K inhibitors. Cancer Cell, 26, 136–149.
MeijerL, BorgneA, MulnerO, ChongJP, BlowJJ, InagakiN, InagakiM, DelcrosJG, MoulinouxJP. (1997) Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent kinases cdc2, cdk2 and cdk5. European Journal of Biochemistry, 243, 527–536.
51.
FryDW, HarveyPJ, KellerPR, ElliottWL, MeadeM, TrachetE, AlbassamM, ZhengX, LeopoldWR, PryerNK, ToogoodPL. (2004) Specific inhibition of cyclin-dependent kinase 4/6 by PD 0332991 and associated antitumor activity in human tumor xenografts. Molecular Cancer Therapeutics, 3, 1427–1438.
ShahMA, SchwartzGK. (2001) Cell cycle-mediated drug resistance: an emerging concept in cancer therapy. Clinical Cancer Research, 7, 2168–2181.
54.
DeepG, AgarwalR. (2008) New combination therapies with cell-cycle agents. Current Opinion in Investigational Drugs, 9, 591–604.
55.
GabrielliB, BrooksK, PaveyS. (2012) Defective cell cycle checkpoints as targets for anti-cancer therapies. Frontiers in Pharmacology, 3, 9.
56.
ScotlandiK, PerdichizziS, ManaraMC, SerraM, BeniniS, CerisanoV, StrammielloR, MercuriM, Reverter-BranchatG, FairclothG, D'IncalciM, PicciP. (2002) Effectiveness of Ecteinascidin-743 against drug-sensitive and -resistant bone tumor cells. Clinical Cancer Research, 8, 3893–3903.
57.
GroharPJ, SegarsLE, YeungC, PommierY, D'IncalciM, MendozaA, HelmanLJ. (2014) Dual targeting of EWS-FLI1 activity and the associated DNA damage response with trabectedin and SN38 synergistically inhibits Ewing sarcoma cell growth. Clinical Cancer Research, 20, 1190–1203.
58.
DemarcqC, BunchRT, CreswellD, EastmanA. (1994) The role of cell cycle progression in cisplatin-induced apoptosis in Chinese hamster ovary cells. Cell Growth & Differentiation, 5, 983–993.
59.
LuGY, HuangSM, LiuST, LiuPY, ChouWY, LinWS. (2014) Caffeine induces tumor cytotoxicity via the regulation of alternative splicing in subsets of cancer-associated genes. International Journal of Biochemistry & Cell Biology, 47, 83–92.
60.
MosmannT. (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65, 55–63.
61.
SiuWY, YamCH, PoonRY. (1999) G1 versus G2 cell cycle arrest after adriamycin-induced damage in mouse Swiss3T3 cells. FEBS Letters, 461, 299–305.
62.
SunM, ZhangN, WangX, CaiC, CunJ, LiY, LvS, YangQ. (2014) Nitidine chloride induces apoptosis, cell cycle arrest, and synergistic cytotoxicity with doxorubicin in breast cancer cells. Tumour Biology, 35, 10201–10212.
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