heptaphylline and 7-methoxyheptaphylline

Heptaphylline derivatives are carbazoles in Clausena harmandiana, a medicinal plant that is utilized for headache, stomach ache, and other treatments of illness. The present study examined the effects of heptaphylline and 7-methoxyheptaphylline on apoptosis of human colon adenocarcinoma cells (HT-29 cell line). Quantification of cell viability was performed using cell proliferation assay (MTT assay) and of protein expression through immunoblotting. The results showed that only heptaphylline, but not 7-methoxyheptaphylline, significantly significantly activated cleaved of caspase-3 and poly (ADP-ribose) polymerase (PARP-1) which resulted in HT-29 cell death. We found that heptaphylline activated BH3 interacting-domain death agonist (Bid) and Bak, proapoptotic proteins. In contrast, it suppressed X-linked inhibitor-of-apoptosis protein (XIAP), Bcl-xL and survivin, inhibitors of apoptosis. In addition, heptaphylline inhibited activation of NF-κB/p65 (rel), a regulator of apoptotic regulating proteins by suppressing the activation of Akt and IKKα, upstream regulators of p65. The findings suggested that heptaphylline induces apoptosis in human colon adenocarcinoma cells .

Topics: Adenocarcinoma; Apoptosis; bcl-2 Homologous Antagonist-Killer Protein; bcl-X Protein; BH3 Interacting Domain Death Agonist Protein; Carbazoles; Caspase 3; Cell Proliferation; Cell Survival; Colonic Neoplasms; HT29 Cells; Humans; I-kappa B Kinase; Inhibitor of Apoptosis Proteins; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerases; Proto-Oncogene Proteins c-akt; Signal Transduction; Survivin; Transcription Factor RelA; X-Linked Inhibitor of Apoptosis Protein

Nineteen carbazole alkaloids modified from heptaphylline (I) and 7-methoxyheptaphylline (II) isolated from Clausena harmandiana were synthesized. Among these derivatives, Ih and IIi showed cytotoxicity against the NCI-H187 cell line with IC(50) values of 0.02 and 0.66 μM, respectively, which are about 138 and 4 fold stronger than the ellipticine standard. In addition, oxime Ih displayed cytotoxicity against KB cells with an IC(50) value of 0.17 μM which is about 10 times stronger than the ellipticine. This compound demonstrated weak cytotoxicity against Vero cells (IC(50) = 66.01 μM). The results show convincingly that Ih may be a promising lead for the development of cytotoxic agents.

Topics: Animals; Carbazoles; Cell Line, Tumor; Chlorocebus aethiops; Drug Screening Assays, Antitumor; Humans; Inhibitory Concentration 50; Magnetic Resonance Spectroscopy; Mass Spectrometry; Spectroscopy, Fourier Transform Infrared; Vero Cells

Four new carbazole alkaloids, claurailas A-D (1-4), as well as 12 known carbazoles and three known coumarins were isolated from the roots of Clausena harmandiana. Heptaphylline (6) and 7-methoxyheptaphylline (7) showed strong cytotoxicity against NCI-H187 and KB cell lines with IC(50) values ranging from 1.3 to 2.7 μM. Compound 7 showed no cytotoxicity against Vero cells.

Topics: Alkaloids; Animals; Antineoplastic Agents, Phytogenic; Carbazoles; Chlorocebus aethiops; Clausena; Coumarins; Drug Screening Assays, Antitumor; Humans; Inhibitory Concentration 50; KB Cells; Plant Roots