ガーナ産寄生植物 Thonningia sanguinea Vahl の成分に関する化学的研究

長崎大学学位論文 [学位記番号]博（医歯薬）甲第1100号 [学位授与年月日]平成30年9月20

An Observational Clinical Study on the Effectiveness of the Aqueous Leaf Extract of Lippia multiflora (Mold) as an Antihypertensive Agent

Hypertension is an issue of public health concern due to its impact on society hence the need to find new treatment options. The leaves of Lippia multiflora (Mold) has traditionally been used in the management of hypertension and stress for more than four decades in Ghana. This study reports on an observational clinical study conducted using twenty-eight (28) hypertensive patients visiting the clinic of the Centre for Scientific Research into Plant Medicine (CSRPM). Participants were asked to prepare an infusion of L. multiflora by boiling 30 g of the plant material in 250 mls of water for four minutes and then filter. The treatment was to be administered twice daily for a period of four (4) weeks. Results showed a significant decline in both systolic blood pressure (SBP) and diastolic blood pressure (DBP) over the period with the difference in mean SBP of 11.6 mmHg and that of the mean DBP recording a 6.43 mmHg decline from the baseline. The aqueous leaf extract of the plant thus holds some prospects for use as an anti-hypertensive agent. Keywords: diastolic BP, hypertension, herbal medicine, Lippia multiflora, observational clinical study, systolic B

Re-evaluating the effi cacy of the aqueous leaf extract of Bridelia ferrugenia and its potential combination with metformin in the management of diabetes mellitus

Herbal medicinal products play a significant role in the management of diabetes and other chronic diseaseseither as a monotherapy or in combination with allopathic treatments. The hypoglycemic activity of a Ghanaianherbal antidiabetic product (Bridelia Tea) prepared from the dried leaves of Bridelia ferrugenia [BRD] was reevaluatedin this study with the view of improving, its activity and explore the possible benefit of using theproduct in combination with metformin (MET). Male and female Sprague-Dawley rats (210-220 g) were renderedhyperglycemic by a single intraperitoneal administration of streptozocin (70 mg/kg) after an overnight fast. Thehyperglycemic rats then received one of the following treatments ([BRD 30 mg/kg or 300 mg/kg], [MET 250 mg/kgor 1000 mg/kg], [MET 250 mg/kg and BRD 30 mg/kg], [MET 1000 mg/kg and BRD 300 mg/kg]). The hypoglycemiceffect of BRD, MET, and the combination of the two products were not significantly different during an initial6 h of monitoring (P > 0.05). In the long-term study over 28 days, BRD 300 mg/kg had hypoglycemic effectscomparable to MET 1000 mg/kg p.o. and better than BDF 30 mg/kg p.o. (P < 0.05). However, the combination ofthe two products BRD and MET at the two doses reduced their therapeutic effect compared to animals receivingeither one of the two treatments alone

Appraisal of the Constituent Plant Materials in a Ghanaian Antifungal Herbal Product; An in vitro Interactive Combination Analysis and a Pilot Clinical Study to Determine Efficacy.

Evidence based use of herbal medicines has a positive implication for both users and society. In this study, component plant materials of a traditional Ghanaian polyherbal skin product comprising Eugenia caryophyllata, Zanthoxylum zanthoxyloides Tridax procumbens, Psidium guajava and Alchornea cordifolia and used in the management of superficial skin infections was evaluated to establish their contribution to the overall therapeutic activity of the product. Each of the five (5) plants was subjected to an in vitro antimicrobial assay using the microtitre broth technique followed by an interactive combination assay for plants demonstrating noteworthy antimicrobial activity (MIC ≤ 1.0 mg/ml). Test strains included Staphyloccocus aureus, Candida albicans, Trichophyton rubrum, Epidermophyton floccusum and Microsporum canis. Eugenia caryophyllata, Zanthoxylum zanthoxyloides and Alchornea cordifolia showed better activity than Psidium guajava, Tridax procumbens and the Total Crude Extract (combination of the 5 extracts). The binary combination of Eugenia caryophyllata and Alchornea cordifolia indicated synergistic and additive activity against all the test strains. An improved biological activity was also observed when a mixture of the two (2) plants at a ratio of Eugenia caryophyllata 60 % (w/w) and Alchornea cordifolia 40 % (w/w) was assayed. A follow up pilot clinical study established that this new recipe was clinically effective but of lower therapeutic effect compared to the original product. In conclusion, the original formulation of the product may be preferred because of the shorter duration of treatment which reduces the risk of harms and cost of treatment

Appraisal of the Constituent Plant Materials in a Ghanaian Antifungal Herbal Product; An in vitro Interactive Combination Analysis and a Pilot Clinical Study to Determine Efficacy.

Evidence based use of herbal medicines has a positive implication for both users and society. In this study, component plant materials of a traditional Ghanaian polyherbal skin product comprising Eugenia caryophyllata, Zanthoxylum zanthoxyloides Tridax procumbens, Psidium guajava and Alchornea cordifolia and used in the management of superficial skin infections was evaluated to establish their contribution to the overall therapeutic activity of the product. Each of the five (5) plants was subjected to an in vitro antimicrobial assay using the microtitre broth technique followed by an interactive combination assay for plants demonstrating noteworthy antimicrobial activity (MIC ≤ 1.0 mg/ml). Test strains included Staphyloccocus aureus, Candida albicans, Trichophyton rubrum, Epidermophyton floccusum and Microsporum canis. Eugenia caryophyllata, Zanthoxylum zanthoxyloides and Alchornea cordifolia showed better activity than Psidium guajava, Tridax procumbens and the Total Crude Extract (combination of the 5 extracts). The binary combination of Eugenia caryophyllata and Alchornea cordifolia indicated synergistic and additive activity against all the test strains. An improved biological activity was also observed when a mixture of the two (2) plants at a ratio of Eugenia caryophyllata 60 % (w/w) and Alchornea cordifolia 40 % (w/w) was assayed. A follow up pilot clinical study established that this new recipe was clinically effective but of lower therapeutic effect compared to the original product. In conclusion, the original formulation of the product may be preferred because of the shorter duration of treatment which reduces the risk of harms and cost of treatment

Thonningia sanguinea Extract: Antioxidant and Cytotoxic Activities Supported by Chemical Composition and Molecular Docking Simulations

The current study was designed to investigate the antioxidant and cytotoxic activities of Thonningia sanguinea whole-plant extract. The total phenolic content was determined using Folin–Ciocalteu reagent and found to be 980.1 mg/g, calculated as gallic acid equivalents. The antioxidant capacity was estimated for the crude extract and the phenolic portion of T. sanguinea, whereupon both revealed a dose-dependent scavenging rate of DPPH• with EC50 values of 36.33 and 11.14 µg/mL, respectively. Chemical profiling of the plant extract was achieved by LC-ESI-TOF-MS/MS analysis, where 17 compounds were assigned, including ten compounds detected in the negative mode and seven detected in the positive mode. The phenolic portion exhibited promising cytotoxic activity against MCF-7 and HepG2 cells, with IC50 values of 16.67 and 13.51 μg/mL, respectively. Phenolic extract treatment caused apoptosis in MCF-7 cells, with total apoptotic cell death 18.45-fold higher compared to untreated controls, arresting the cell cycle at G2/M by increasing the G2 population by 39.7%, compared to 19.35% for the control. The apoptotic investigation was further validated by the upregulation of proapoptotic genes of P53, Bax, and caspases-3,8 9, and the downregulation of Bcl-2 as the anti-apoptotic gene. Bcl-2 inhibition was also virtualized by good binding interactions through a molecular docking study. Taken together, phenolic extract exhibited promising cytotoxic activity in MCF-7 cells through apoptosis induction and antioxidant activation, so further fractionation studies are recommended for the phenolic extract for specifying the most active compound to be developed as a novel anti-cancer agent

GC-MS/MS Quantification of EGFR Inhibitors, <i>β</i>-Sitosterol, Betulinic Acid, (+) Eriodictyol, (+) Epipinoresinol, and Secoisolariciresinol, in Crude Extract and Ethyl Acetate Fraction of <i>Thonningia sanguinea</i>

Medicinal plants are widely used in folk medicine to treat various diseases. Thonningia sanguinea Vahl is widespread in African traditional medicine, and exhibits antioxidant, antibacterial, antiviral, and anticancer activities. T. sanguinea is a source of phytomedicinal agents that have previously been isolated and structurally elucidated. Herein, gas chromatography combined with tandem mass spectrometry (GC-MS/MS) was used to quantify epipinoresinol, β-sitosterol, eriodictyol, betulinic acid, and secoisolariciresinol contents in the methanolic crude extract and its ethyl acetate fraction for the first time. The ethyl acetate fraction was rich in epipinoresinol, eriodictyol, and secoisolariciresinol at concentrations of 2.3, 3.9, and 2.4 mg/g of dry extract, respectively. The binding interactions of these compounds with the epidermal growth factor receptor (EGFR) were computed using a molecular docking study. The results revealed that the highest binding affinities for the EGFR signaling pathway were attributed to eriodictyol and secoisolariciresinol, with good binding energies of −19.93 and −16.63 Kcal/mol, respectively. These compounds formed good interactions with the key amino acid Met 769 as the co-crystallized ligand. So, the ethyl acetate fraction of T. sanguinea is a promising adjuvant therapy in cancer treatments