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Eight new annonaceous acetogenins, squamotin A-D (1-4), annosquatin IV-V (5 and 6), muricin O (7) and squamosten B (8), together with four known ones (9-12) were isolated from the seeds of Annona squamosa. Their structures were elucidated by chemical methods and spectral data. The inhibitory activities of compound 1-9 against three multidrug resistance cell lines were evaluated. All tested compounds showed strong cytotoxicity.
Subject(s)
Humans , Acetogenins , Chemistry , Pharmacology , Toxicity , Annona , Chemistry , Antineoplastic Agents, Phytogenic , Chemistry , Pharmacology , Toxicity , Cell Line, Tumor , Cell Survival , Drug Resistance, Neoplasm , Drug Screening Assays, Antitumor , Molecular Structure , Plant Extracts , Chemistry , Pharmacology , Toxicity , Seeds , ChemistryABSTRACT
Annonaceous acetogenins (ACGs) are effective part extracted and separated from Annona squamosa seeds, they have good antitumor activity against a variety of tumor cells. However, the solubility of ACGs is poor with serious toxic and side effects, which greatly limits their application in clinical practice. In this study poloxamer 188 (P188) was selected as a drug carrier or a stabilizer to prepare ACGs nanosuspensions (ACGs-NSps) using anti-solvent precipitation. The nanosuspensions were examined via dynamic light scattering (DLS) method to examine size of the nanosuspensions. Transmission electron microscopy was used to observe their morphology. HPLC assay was used to measure their drug loading content and the in vitro drug release. The stability of ACGs-NSps at room temperature, in various physiological media and plasma, and the hemolytic test and lyophilization were all investigated. MTT assay was performed to study the cytotoxocity of ACGs-NSps against four tumor cell lines. 4T1 bearing tumor model was used to assess their in vivo antitumor therapeutic efficacy. The obtained ACGs-NSps were spherical, the average particle size was 169.4±1.25 nm, the polydispersity index (PDI) value was 0.130±0.020, the zeta potential was -19.8 mV and the drug loading content was 48.18%. ACGs-NSps were stable at room temperature for at least 15 days. They could be lyophilized in the presence of 0.5% glucose and 2.0% P188. ACGs-NSps showed sustained in vitro drug release, and the cumulative drug release reached 80.82% within 144 hours. ACGs-NSps maintained their particle size in various physiological media, and plasma with no hemolysis and then met demands of both oral and intravenous administration. In contrast to free ACGs, ACGs-NSps displayed significantly higher cytotoxicity against 4T1 (IC50, 0.892±0.124 μg·mL-1 vs 2.495±0.108 μg·mL-1, P 50, 0.747±0.051 μg·mL-1 vs 2.204±0.064 μg·mL-1, P 50, 2.265±0.081 μg·mL-1 vs 4.159±0.071 μg·mL-1, P 50, 0.473±0.024 μg·mL-1 vs 1.196±0.022 μg·mL-1, P in vivo study demonstrated that the daily oral administration of ACGs-NSps (3 mg·kg-1) resulted in higher tumor inhibition rate compared to ACGs/oil solution (67.23% vs 53.11%), comparable to the intravenous injection of 0.5 mg·kg-1 ACGs-NSps every other day (70.34%). Nanosuspensions effectively solved the problem of ACGs insolubility and difficulty in drug delivery. Using P188, a pharmaceutic adjuvant approved by FDA for iv injection, the resultant ACGs-NSps appear promising as an anti-tumor drug that can be used in clinic.
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Objective To study the effect of annonaceous acetogenins (ACGs) on human gastric cancer cells in vitro. Methods After ACGs were administered to gastric cancer cells in vitro, the cell viability, cell adhesion ability and cell migration ability were assessed by MTT assay, adhesion assay and wound-healing assay, respectively. Results ACGs inhibited the cell viability, adhesion ability and migration ability in a dose-dependent manner in gastric cancer cells. Conclusion ACGs could inhibit cell activities of human gastric cancer cells in viro, and will be developed as a promising anticancer candidate and used in gastric cancer.
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The present research was launched to investigate the effects of 12 annonaceous acetogenins (ACGs) on human hepatic carcinoma cell line SMMC-7721/ADR, and to find out their structure-activity relationship. SMMC-7721/ADR cells were treated with 12 ACGs including annosquamin A(1)，annosquamin B(2)，annotemoyin-1(3)，uvariamicin Ⅱ(4)，annosquacin D(5)，annosquacin B(6)，isodesacetyluvaricin(7)，uvarigrandin A(8)，squamostatin D(9)，squamostatin E(10)，squamostatin A(11)，and 12,15-cis-squamostatin A(12) for 24 h, and the different expression of the target gene NDUFV2 were detected by quantitative real-time PCR. All the tested compounds made the expression of the target gene NDUFV2 decreased on human hepatic carcinoma cell line SMMC-7721/ADR, of which the bistetrahydrofuran ACGs showed the best activity,which the non-adjacent bistetrahydrofuran ACGs displayed the worst activity.The ACGs with the reducing number of carbons between γ-unsaturated lactone and the close tetrahydrofuran (THF) ring are more potent. For bistetrahydrofuran ACGs with the same nucleus skeleton,they would be more active as more hydroxyls on aliphatic chain, which for the non-adjacent bistetrahydrofuran ACGs with less hydroxyls on aliphatic chain that would be more active. ACGs with 3 hydroxyls on aliphatic chain would be more active. ACGs with threo configuration are more active than erythro configurotion, and the compounds with cis THF ring seem to be superior to those of trans THF ring. Furthermore, the ACGs with the reducing number of carbons between terminal methyl and the close tetrahydrofuran (THF) ring are more potent.
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10 kinds of annonaceous acetogenins were selected for antitumor activity testing against human lung cancer cell line A549/Taxol and the structure activity relationship was analyzed.MTT assay was used to detect the inhibitory activities of 10 kinds of annonaceous acetogenins and positive drugs against A549/Taxol cells, respectively uvariamicin-Ⅲ(1), uvariamicin-Ⅱ(2), annosquacin D(3), desacetyluvaricin(4), annosquatin A(5), squamostatin D(6), bullatacin(7), squamocin(8), motrilin(9), annosquatin B(10), verapamil and cisplatin. Annonaceous acetogenins showed significant inhibitory activities against A549/Taxol cells, and were more potent than the positive drug verapamil and cisplatin.The more carbon atoms between the tetrahydrofuran ring and the lactone ring of annonaceous acetogenins exhibited more potency.Besides,ACGs with two substituted hydroxyl showed more potency than the compounds with three substituted hydroxyl in the bis-adjacent-THF ACGs. Furthermore, ACGs with three substituted hydroxyl showed more potency than the compounds with four substituted hydroxyl among the no bis-adjacent-THF ACGs.
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<p><b>OBJECTIVE</b>To confirm the anticancer effect of total annonaceous acetogenins (TAAs) abstracted from Annona squamosa Linn. on human hepatocarcinoma.</p><p><b>METHODS</b>The inhibitory effect of TAAs was demonstrated in H22-bearing mice. The potency of TAAs was confirmed as its 50% inhibiting concentration (IC50) on Bel-7402 cell under Sulfur Rhodamine B staining. Both underlying mechanisms were explored as cellular apoptosis and cell cycle under flow cytometry. Mitochondrial and recipient apoptotic pathways were differentiated as mitochondrial membrane potential under flow cytometry and caspases activities under fluorescence analysis.</p><p><b>RESULTS</b>The inhibitory rate of TAAs in mice was 50.98% at 4 mg/kg dose. The IC50 of TAAs on Bel-7402 was 20.06 µg/mL (15.13-26.61µg/mL). Effective mechanisms of TAAs were confirmed as both of arresting cell cycle at G1 phase and inducing apoptosis dose- and time-dependently. Mitochondrial and recipient pathways involved in apoptotic actions of TAAs.</p><p><b>CONCLUSION</b>TAAs is effective for hepatocarcinoma, via inhibiting proliferation and inducing apoptosis.</p>
Subject(s)
Animals , Humans , Male , Mice , Acetogenins , Chemistry , Pharmacology , Therapeutic Uses , Annona , Chemistry , Antineoplastic Agents, Phytogenic , Chemistry , Pharmacology , Therapeutic Uses , Apoptosis , Carcinoma, Hepatocellular , Drug Therapy , Pathology , Caspases , Metabolism , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Chromatography, High Pressure Liquid , Dose-Response Relationship, Drug , Liver Neoplasms , Drug Therapy , Pathology , Membrane Potential, Mitochondrial , Organ Specificity , Spleen , Thymus Gland , Xenograft Model Antitumor AssaysABSTRACT
Objective: To investigate the effects of 15 annonaceous acetogenins (ACGs) on human breast cancer cell line MCF-7/ADR, and to find out their structure-activity relationship. Methods: MCF-7/ADR cells were treated with 15 ACGs such as annotemoyin-1 (1), annosquamin B (2), annosquamin A (3), annosquamin C (4), annosquacin C (5), urarigrandin A (6), isodesacetyluvaricin (7), annosquacin D (8), annosquacin B (9), 12, 15-cis-squamostatin-A (10), squamostatin-A (11), squamostanin-B (12), squamostanin-A (13), squamostatin-D (14), and squamostatin-E (15) for 48 h, and the inhibition on MCF-7/ADR cells was detected using MTT assay. Results: All the tested compounds showed significant inhibitory activities against MCF-7/ADR cells, and were more potent than the standard control verapamil. The activity of compound 1 was 190 times higher than that of verapamil. Conclusion: The ACGs with more carbons between tetrahydrofuran (THF) ring and γ-unsaturated lactone are more potent. If all other structural features are identical, the ACGs with more hydroxyls on aliphatic chain would be more active, and four hydroxyls might be optimal among bis-nonadjacent-THF ACGs. Moreover, ACGs with stereochemical arrangement of erythro are more active than those of threo, and the compounds with THF ring configuration of cis seem to be superior to those of trans. Furthermore, bis-adjacent-THF ACGs with molecular weight of 622 and with three hydroxyl groups and stereochemical arrangement of erythro partly produce notable cytotoxicity.
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Objective To investigate the apoptosis induction of Bullatacin on A549 cell line of pulmonary adenocarcinoma. Methods The MTT assay was used to detect the growth inhibition rates of A549 cells cultured with Bullatacin in different concentrations (6.25, 12.5, 25, 50, 100μg/mL). 25μg/mL Bullatacin was used to culture A549 cells for 0, 12, 24, 48 h. The cell cycle distribution and apoptosis were measured by flow cytemetry. The protein expressions of ERK, JNK, and p38 were studied by Western blot. Results Dosage dependence was obviously showed after the different concentrations of Bullatacin were used to A549, and 25 μg/mL;Bullatacin blocked A549 cell in G0/G1 periods and induced its apoptosis. Compared with the blank group, protein expressions of P-ERK, P-JNK, and P-p38 were all increased by different degrees. Conclusion Bullatacin significantly inhibits the proliferation and induces the apoptosis of A549 cell. Its mechanism is related to activity of MAPK pathway thought the phosphorylation of the three protein kinases by Bullatacin.
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Objective: To investigate the activity of mono-tetrahydrofuran (THF) annonaceous acetogenins (ACGs) against mitochondrial complex I of rats. Methods: The inhibitory activity of mono-THF ACGs with six different chemical structures against mitochondrial complex I of rats was investigated to clarify the carbon number and substituted hydroxyl number between THF ring and lactone ring as well as the effect of the core configuration in THF ring on mitochondrial complex I of rats. Results: The results show that mono-THF ACGs can inhibit the mitochondrial complex I of rats. With analysis of the results from the structure-activity relationship between antitumoral activity and their chemical structure of mono-THF ACGs, the less the carbon number between the two rings is, the better their inhibitory activities are; The number of substituted hydroxyl groups is not the decisive factor for influencing its activity in mono-THF ACGs. Conclusion: The inhibitory activity of compound's configurations with th/t/er is better than that of the compound's configurations with th/t/th in mono-THF ACGs.
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Objective To observe the inhibitory action of annonaceous acetogenins AS-4 on human liver cancer cell line HepG2 in vitro. Method Using different concentration of annonaceous acetagenins group(6.25,12.5,25,50 ?g/mL),control group(25 ?g/mL) of masculine(DDP),control blank group,after routine culture of 24 or 48 hours,the effect of annonaceous acetogenins AS-4 on HepG2 was analyzed by colorimetry assay(MTT) and inverted phase-contrast microscope observing. Result AS-4 had the best inhibitory effect on HepG2,the highest inhibitory rate was 91.68%,and the effect was time-dosage dependent,and the 48 h IC50 was 6.67 ?g/mL. Conclusion AS-4 has lethal effect on HepG2 in vitro,and shows obvious cytotoxic effect.
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A large number of natural acetogenins have been isolated from se veral genera of the plant family Annonaceae. Many of these compounds have very p otent and diverse biological effects such as cytotoxic, antitumour, anti-malari al, pesticidal and anti-feedant activities. This paper reviews research finding s about the effects of annonaceous acetogenins on antitumor, MDR, mechanism of ac tion, structural-activity relationships etc.