Chemical and biological evaluation of metabolites isolated from fruits, wastes and endophytic fungi of cultivated Ammi majus L. plant / by Noha Mostafa Fathallah Fathi (Assistant Lecturer of Pharmacognosy and Medicinal Plants, Faculty of Pharmaceutical Sciences and Pharmaceutical Industries, Future University)

Supervision of Prof. Dr. Mostafa Aly Abd El Kawy (Professor of Pharmacognosy, Faculty of Pharmacy, Cairo University), Prof. Dr. Osama Mostafa Salama (Professor of Pharmacognosy, Faculty of Pharmaceutical Sciences and Pharmaceutical Industries, Vice President Future University, Egypt), Associate Prof. Marwa Yousry Mostafa Issa (Associate professor of Pharmacognosy, Faculty of Pharmacy, Cairo University, Egypt), Dr. Mokhtar Mohamed Bishr (Plant General Manager and Technical Director, Arab Company for Pharmaceuticals and Medicinal Plants, Egypt)

Thesis (P.hD.)-Cairo university, Faculty of pharmacy, Department of Pharmacognosy, 2021.

Includes bibliographical references.

Ammi majus L., family Apiaceae growing in Egypt is famous for comprising active constituents as coumarins, flavonoids, and steroids. Its main used part is the fruits that contain furanocoumarins widely used in skin conditions like vitiligo and psoriasis. Firstly: This study is meant to investigate the bioactive metabolites present in the aerial parts (waste) in comparison with that of the fruits and optimize the cultivation conditions to enhance the constituents present in the fruits. Optimization was via exposure to drought stress and applying an exogenous phytohormone (ethephon) then comparing the results with that of the control fruit group. The physical parameters besides the differences in the total phenolics, total flavonoid contents, and the antioxidant activity of the extracts were evaluated. Remarkably, the aerial parts revealed higher total phenolics and flavonoids with much stronger antioxidant activity than the fruit extract. The positive and negative metabolic profiling using UPLC/MS-MS of the aerial parts extract revealed different compounds when compared to the compounds present in the fruits. Secondly: The Ammi majus, fruits were investigated for their phytochemical compounds responsible for their pharmacological activities. The dried powdered plant was extracted with methanol, fractionated using gradient elution then the CH2Cl2 fraction was purified using puri-flash. The compounds isolated were identified using 1HNMR and 13CNMR as β-sitosterol, Isoarnottinin, Xanthotoxin, Marmesin, Imperatorin, and Ammirin. Thirdly: An endophytic fungus was isolated from the fruits and identified as Aspergillus amstelodami (MK215708). After mass production of the fungus on rice media, the active metabolites were extracted using EtOAc then fractionated and purified using preparative HPLC to give Dihydroauroglaucin, tetrahydroauroglaucin, 2-(3,6-dihydroxyhepta-1,4-dien1-yl)-3,6-dihydroxy-5-(dimethylallyl) benzaldehyde, isotetrahydroauroglaucin), flavoglaucin and, 2-(2, 3-epoxy-1,3,5- heptatrienyl)-6-hydroxy-5-(3-methyl-2-butenyl) benzaldehyde. Structure elucidation was carried out using (1 H-and 13 C-NMR, HMBC, HSQC, and COSY). Finally, Antimicrobial, Antibiofilm, cytocompatibility, and cytotoxicity assays were done based on biological guided technique. The antimicrobial and antibiofilm results revealed that the major fungus compound (dihydrauroglaucin) had the strongest activity followed by the fungus fractions, the FEA then the plant extract. The cytocompatibility effect of AME, FEA, and dihydrauroglaucin revealed no toxicity on the normal human fibroblastic cell line (BHK). For cytotoxicity against HepG2, the AME, FEA, their fractions, and the major plant compound xanthotoxin were investigated. Among all the tested samples, the xanthotoxin revealed the most potent cytotoxic activity with an IC50 of 6.9 μg/ ml.

Text in English, abstracts in English and Arabic.