Anticholangiocarcinoma activity and toxicity of the Kaempferia galanga Linn. Rhizome ethanolic extract

Abstract

BackgroundCholangiocarcinoma (CCA) is an important public health problem in several tropical and subtropical parts of the world particularly Thailand. Chemotherapy of CCA is largely ineffective and discovery and development of effective alternative drugs is urgently needed. The objective of the study was to confirm the anti-CCA potential as well as toxicity of the crude extract of Kaempferia galangal Linn. (rhizome) both in vitro and in animal models.MethodsThe ethanolic extract of K. galanga Linn. rhizome, ethyl-p-methoxycinnamate (EPMC) and 5-fluorouracil (5-FU) were evaluated for their cytotoxic activities against CCA cell line (CL-6) using MTT cell proliferation assay. Acute and subacute toxicity of the extract were evaluated in ICR (Imprinting Control Region) mice according to the OECD (International Organization for Economic Co-operation and Development) Guideline. Anti-CCA activity was evaluated in CCA- xenografted nude mice.ResultsResults of cytotoxicity test showed moderate activity of the extract and EPMC with median (95% confidence interval: 95% CI) 50% inhibitory concentration (IC50) of 64.2 (57.76–72.11) and 49.19 (48.16–52.29) μg/ml, respectively. The IC50 of 5-FU was 107.1 (103.53–109.64) μg/ml. The selectivity index (SI) values for the extract, EPMC and 5-FU against human normal cell line (OUMS) and cancer cell line (CL-6) were 2.2, 2.09 and 1.31, respectively. Toxicity testing revealed no overt toxic effect up to the maximum single oral dose of 5000 mg/kg body weight and up to daily dose of 1000 mg/kg body weight for 30 days. The extract at the maximum tolerated dose level of 1000 mg/kg body weight for 30 days exhibited promising anti-CCA activity in CL6-xenografted nude mice as determined by inhibitory activity on tumor growth (58.41%) and lung metastasis (33.3%), as well as prolongation of survival time (62 days).ConclusionThe K. galangal Linn. rhizome extract and its bioactive compound EPMC exhibited moderate cytotoxic activity against human CCA tumor (CL-6) cell line. Results of toxicity testing suggest that the extract was well tolerated up to the maximum single oral dose of 5000 mg/kg body weight and daily dose of 1000 mg/kg body weight for 30 days. The extract exhibited promising anti-CCA activity in CL6-xenografed nude mice as determined by significant inhibitory activity on tumor growth and lung metastasis, as well as prolongation of survival time.

Cytotoxicity, Cell Cycle Arrest, and Apoptosis Induction Activity of Ethyl-p-methoxycinnamate in Cholangiocarcinoma Cell

“…The selectivity to CL-6 cell was 3.7 times compared to a normal cell, OUMS-36T-1F (Table 1). It is noted for the variable activity, and selectivity of EPMC reported in this study compared with the previous study against the same cell lines (Amuamuta, et al, 2017). In the previous study, the IC 50 (median [95% CI]) and selectivity index of EPMC were shown to be 49.19 (48.16-52.29) μg/ml and 2.09, respectively.…”

“…In the previous study, the IC 50 (median [95% CI]) and selectivity index of EPMC were shown to be 49.19 (48.16-52.29) μg/ml and 2.09, respectively. The potency of activity of this active compound was about 1.3 times of the crude ethanolic extract (median IC 50 64.2 µg/ml) (Amuamuta, et al, 2017). The anticancer activity of EPMC could be improved when combined with other compounds as the components of K. galanga L. such as kaempferol or kaempferide.…”

“…This suggests the potential role of the compound to interrupt cell migration and metastasis. Results of the previous toxicity study showed that K. galangal L. extract exhibited promising anti-cholangiocarcinoma activity in CL6-xenografted nude mice as determined :10.31557/APJCP.2020.21.4.927 Cytotoxicity, Cell Cycle Arrest, and Apoptosis of EPMC in CL-6 by significant inhibitory activity on tumor growth and lung metastasis, as well as prolongation of survival time (Amuamuta, et al, 2017).…”

“…The potential anticancer activity of K. galanga L. crude extract has previously been reported in the CL-6 cell (Amuamuta et al, 2017;Mahavorasirikul et al, 2010). However, the study about the anticancer activity of the isolated compound, EPMC, in cholangiocarcinoma cell still limited.…”

Anti‐inflammatory effect of isopimarane diterpenoids from Kaempferia galanga

“…Its essential oil is used in cosmetic, perfumery, and incense industry (Duke, Bogenschutz‐Godwin, duCellier, & Duke, ; Ibrahim, ; Kanjanapothi et al, ; Parvez, Khan, Islam, & Hasan, ). K. galanga powder, extracts, and constituents have been reported to operate in many biological activities such as the antiangiogenic activity (He, Yue, Lau, Ge, & But, ), anthelmintic activity (Kiuchi, Nakamura, Tsuda, Kondo, & Yoshimura, ), anti‐inflammation against carrageenan‐induced rat paw edema (Ridtitid, Saewong, Reanmongkol, & Wongnawa, ; Umar, Bin Asmawi, Sadikun, Altaf, & Iqbal, ), anti‐inflammation against IL‐6 production (Aroonrerk & Kamkaen, ), antimicrobial and antifungal (Tewtrakul, Yuenyongsawad, Kummee, & Atsawajaruwan, ), anticholangiocarcinoma activity (Amuamuta, Plengsuriyakarn, & Na‐Bangchang, ), wound healing activity in rat model (Shanbhag et al, ), and so forth. Constituents found in volatile oil are trans‐ethyl‐ p ‐methoxycinnamate, trans‐ethyl cinnamate, carvone, 1,8‐cineole, 3‐carene, borneol, camphene, pinene, eucarvone, germacrene, α‐gurjunnene, kaempferol, kaempferide, pentadecane, and so forth (Indrayan, Kurian, Tyagi, Shatru, & Rathi, ; Raina & Abraham, ; Tewtrakul et al, ).…”