Ethnobotanical study of anticancer plants used in Ugbine, Nigeria and bioactivity-guided isolation of cytotoxic agents from Synclisia scabrida (Miers) ex oliv
Introduction: Plants are promising source of novel anticancer agents. Although many breakthroughs have been made in cancer treatment, it remains suboptimal. Therefore, alternative therapeutics with novel mechanisms of action, with minimal side effects and costs are urgently needed. Method: Struc...
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my-upm-ir.1038092023-04-18T03:24:26Z Ethnobotanical study of anticancer plants used in Ugbine, Nigeria and bioactivity-guided isolation of cytotoxic agents from Synclisia scabrida (Miers) ex oliv 2022-06 Ngozi, Nwaefulu Ogochukwu Introduction: Plants are promising source of novel anticancer agents. Although many breakthroughs have been made in cancer treatment, it remains suboptimal. Therefore, alternative therapeutics with novel mechanisms of action, with minimal side effects and costs are urgently needed. Method: Structured questionnaires were used for the ethnobotanical study. The ground whole roots and leaves of Synclisia scabrida (Miers) ex Oliv (SS) and leaves of Petiveria alliacea L. (PA) were sequentially and separately extracted using a solid–solvent ratio of 1:2, of hexane, dichloromethane, ethyl acetate, methanol, or dichloromethane: methanol (1:1), by cold maceration for 72 h followed by another cold maceration for 24 h. The extract was filtered, concentrated at 40°C with a rotary evaporator and further dried in the oven set at 40°C to obtain crude extracts, which were stored in a refrigerator. A preliminary phytochemical analysis of SS whole root methanol extract was carried out. MTT cell viability assay was employed to evaluate the cytotoxicity potential of the extracts/compounds in colon (HCT-116), prostate (PC-3), breast (MCF-7), pancreatic (PANC-1) cancer cells and human normal lung (BEAS-2B) and murine microglial (BV- 2) cells. Cell cycle distribution of treated PANC-1 was analysed by flow cytometry for understanding of the mechanism behind the observed anticancer potential of SS. Using a bioassay-guided approach, the most active extract/fraction were purified to yield 4 active compounds (SS_C1, SS_C2, SS_C3 and SS_C4). The structures of SS_C2 and SS_C4 were successfully characterised using 1H, 13C-NMR, COSY, HMBC, HSQC, FTIR and LC/MS Q-TOF spectroscopy. The compounds’ molecular targets were predicted using in silico molecular docking and molecular dynamic approaches. Results: The ethnobotanical study revealed the use of SS and PA for cancer treatment. Phytochemical screening of the methanol extract confirmed the presence of alkaloids, tannins, flavonoids, steroids, and fixed oil. The methanol extract had the following IC50 (μg/mL) in cancer cell lines: HCT-116 (23.3 ± 10.4), MCF-7 (35.0 ± 5.0), PC-3 (29.3 ± 11.0) and PANC-1 (5.7 ± 1.2). SS induced a dose-dependent arrest of PANC-1 cells in the S phase. Structure elucidation of SS_C2 and SS_C4 revealed they are bisbenzylisoquinoline alkaloids. Both compounds were identified as close analogues of cosculine and cycleanine, respectively. SS_C2 (IC50 = 46.7 ± 5.8 μg/mL; SI = 2.1; P<0.000) and SS_C4 (IC50 = 41.7 ± 7.6 μg/mL; SI = 2.4; P<0.000) were selectively toxic towards HCT-116 and MCF-7, respectively. The selectivity was far greater than that of gemcitabine, which showed higher toxicity to normal lung cells (BEAS-2B). The in silico molecular dynamic simulations results showed RMSF plots of PARP1–ligand complexes were more stable than the apo protein. This shows a very high PARP1-ligand complexation stability, suggesting that both ligands (SS_C2 and SS_C4) are potential PARP1 inhibitors. Conclusion: SS_C1 was active against colon and pancreatic cancers, SS_C2 was selectively active against colon cancer while SS_C4 had the best selectivity towards breast cancer cells. SS_C2 and SS_C4 are new bisbenzylisoquinoline alkaloids, and potential PARP1 inhibitors which could become leads for the development of targeted therapy for the treatment of colon, breast, and pancreatic cancers. Ethnobotany Plants, Medicinal 2022-06 Thesis http://psasir.upm.edu.my/id/eprint/103809/ http://psasir.upm.edu.my/id/eprint/103809/1/NWAEFULU%20OGOCHUKWU%20-%20IR.pdf text en public doctoral Universiti Putra Malaysia Ethnobotany Plants, Medicinal Stanslas, Johnson |
| institution |
Universiti Putra Malaysia |
| collection |
PSAS Institutional Repository |
| language |
English |
| advisor |
Stanslas, Johnson |
| topic |
Ethnobotany Ethnobotany |
| spellingShingle |
Ethnobotany Ethnobotany Ngozi, Nwaefulu Ogochukwu Ethnobotanical study of anticancer plants used in Ugbine, Nigeria and bioactivity-guided isolation of cytotoxic agents from Synclisia scabrida (Miers) ex oliv |
| description |
Introduction: Plants are promising source of novel anticancer agents. Although many
breakthroughs have been made in cancer treatment, it remains suboptimal. Therefore,
alternative therapeutics with novel mechanisms of action, with minimal side effects and
costs are urgently needed.
Method: Structured questionnaires were used for the ethnobotanical study. The ground
whole roots and leaves of Synclisia scabrida (Miers) ex Oliv (SS) and leaves of Petiveria
alliacea L. (PA) were sequentially and separately extracted using a solid–solvent ratio
of 1:2, of hexane, dichloromethane, ethyl acetate, methanol, or dichloromethane:
methanol (1:1), by cold maceration for 72 h followed by another cold maceration for 24
h. The extract was filtered, concentrated at 40°C with a rotary evaporator and further
dried in the oven set at 40°C to obtain crude extracts, which were stored in a refrigerator.
A preliminary phytochemical analysis of SS whole root methanol extract was carried
out. MTT cell viability assay was employed to evaluate the cytotoxicity potential of the
extracts/compounds in colon (HCT-116), prostate (PC-3), breast (MCF-7), pancreatic
(PANC-1) cancer cells and human normal lung (BEAS-2B) and murine microglial (BV-
2) cells. Cell cycle distribution of treated PANC-1 was analysed by flow cytometry for
understanding of the mechanism behind the observed anticancer potential of SS. Using
a bioassay-guided approach, the most active extract/fraction were purified to yield 4
active compounds (SS_C1, SS_C2, SS_C3 and SS_C4). The structures of SS_C2 and
SS_C4 were successfully characterised using 1H, 13C-NMR, COSY, HMBC, HSQC,
FTIR and LC/MS Q-TOF spectroscopy. The compounds’ molecular targets were
predicted using in silico molecular docking and molecular dynamic approaches.
Results: The ethnobotanical study revealed the use of SS and PA for cancer treatment.
Phytochemical screening of the methanol extract confirmed the presence of alkaloids,
tannins, flavonoids, steroids, and fixed oil. The methanol extract had the following IC50
(μg/mL) in cancer cell lines: HCT-116 (23.3 ± 10.4), MCF-7 (35.0 ± 5.0), PC-3 (29.3 ±
11.0) and PANC-1 (5.7 ± 1.2). SS induced a dose-dependent arrest of PANC-1 cells in
the S phase. Structure elucidation of SS_C2 and SS_C4 revealed they are
bisbenzylisoquinoline alkaloids. Both compounds were identified as close analogues of
cosculine and cycleanine, respectively. SS_C2 (IC50 = 46.7 ± 5.8 μg/mL; SI = 2.1;
P<0.000) and SS_C4 (IC50 = 41.7 ± 7.6 μg/mL; SI = 2.4; P<0.000) were selectively toxic
towards HCT-116 and MCF-7, respectively. The selectivity was far greater than that of
gemcitabine, which showed higher toxicity to normal lung cells (BEAS-2B). The in
silico molecular dynamic simulations results showed RMSF plots of PARP1–ligand
complexes were more stable than the apo protein. This shows a very high PARP1-ligand
complexation stability, suggesting that both ligands (SS_C2 and SS_C4) are potential
PARP1 inhibitors.
Conclusion: SS_C1 was active against colon and pancreatic cancers, SS_C2 was
selectively active against colon cancer while SS_C4 had the best selectivity towards
breast cancer cells. SS_C2 and SS_C4 are new bisbenzylisoquinoline alkaloids, and
potential PARP1 inhibitors which could become leads for the development of targeted
therapy for the treatment of colon, breast, and pancreatic cancers. |
| format |
Thesis |
| qualification_level |
Doctorate |
| author |
Ngozi, Nwaefulu Ogochukwu |
| author_facet |
Ngozi, Nwaefulu Ogochukwu |
| author_sort |
Ngozi, Nwaefulu Ogochukwu |
| title |
Ethnobotanical study of anticancer plants used in Ugbine, Nigeria and bioactivity-guided isolation of cytotoxic agents from Synclisia scabrida (Miers) ex oliv |
| title_short |
Ethnobotanical study of anticancer plants used in Ugbine, Nigeria and bioactivity-guided isolation of cytotoxic agents from Synclisia scabrida (Miers) ex oliv |
| title_full |
Ethnobotanical study of anticancer plants used in Ugbine, Nigeria and bioactivity-guided isolation of cytotoxic agents from Synclisia scabrida (Miers) ex oliv |
| title_fullStr |
Ethnobotanical study of anticancer plants used in Ugbine, Nigeria and bioactivity-guided isolation of cytotoxic agents from Synclisia scabrida (Miers) ex oliv |
| title_full_unstemmed |
Ethnobotanical study of anticancer plants used in Ugbine, Nigeria and bioactivity-guided isolation of cytotoxic agents from Synclisia scabrida (Miers) ex oliv |
| title_sort |
ethnobotanical study of anticancer plants used in ugbine, nigeria and bioactivity-guided isolation of cytotoxic agents from synclisia scabrida (miers) ex oliv |
| granting_institution |
Universiti Putra Malaysia |
| publishDate |
2022 |
| url |
http://psasir.upm.edu.my/id/eprint/103809/1/NWAEFULU%20OGOCHUKWU%20-%20IR.pdf |
| _version_ |
1776100366406909952 |