Phytochemical, In Vitro Biological, And Computational Analyses Of Bioactive Compounds From Leaf Extracts Of Some Selected Ethiopian Medicinal Plants

Abstract

The Relationship Between Humans And Herbal Medicinal Plants Has A Long History. Nowadays, Interest In Medicinal And Aromatic Plants Is Increasing Due To Rising Consumer Demand And Their Various Culinary And Medicinal Applications. This Trend Is Fueled By Challenges Such As Antimicrobial Resistance And The Emergence Of Opportunistic Pathogens, Which Complicate The Treatment Of Infectious Diseases. In Addition, Cancer Remains A Significant Research Focus Because Of Its Considerable Impact On Public Health. In Ethiopia, There Is A Notable Gap In Addressing Herbal Medicines For Drug-Resistant Diseases And Cancers, Making This A Critical Issue In Health Biotechnology Research Today. Successively, The Primary Aim Of This Research Is To Explore The Phytochemicals And Assess The In-Vitro Antimicrobial And Anticancer Activities Of The Leaves Extracts Obtained By Maceration Method Of Vernonia Amygdalina, Otostegia Integrifolia, And Salvia Rosmarinus, Which Are Among The Most Commonly Used Medicinal Plants By Traditional Healers. The Essential Oils (Eos) Obtained From The Leaves Of V. Amygdalina, O. Integrifolia, And S. Rosmarinus By Hydro-Distillation Method Were Analyzed By Gc-Ms. In Vitro Techniques Were Employed To Evaluate Various Biological Activities, Including Antimicrobial, Anticancer, Antioxidant, Anti-Inflammatory, Anti-Protozoa, And Antiviral Effects. This Was Accomplished Using Different Assays: Agar Disc Diffusion For Assessing Antimicrobial Activity, Methylthiazolyl Diphenyl-Tetrazolium Bromide (Mtt) Bioassay For Anticancer Effects, And Diphenyl-1- Picrylhydrazyl (Dpph) Bioassay For Measuring Antioxidant Activity. Additionally, The Inhibition Of Albumin Denaturation And Proteinase Was Measured For Antiinflammatory Activity, While Broth Inhibition On Peptone-Yeast Extract-Glucose (Pyg) Bioassay For Assessing Anti-Protozoa Effects And Micro-Inhibition Assays On Madin-Darby Canine Kidney (Mdck) Cells Were Performed To Evaluate Antiviral Effects. The Anti-Proliferative Activities Of Three Plant Extracts Were Examined Against Hela Cells, Which Are Continuously Cultured. For The In Silico Analysis, Drug-Likeness And Absorption, Distribution, Metabolism, And Excretion ?�? Toxicity (Admet) Profiling Were Assessed Using Biovia-2020 Software, Which Facilitated Molecular Docking Tests On The Compounds Under Investigation. Statistical Analysis Was Conducted Using Graphpad Prism Version 8.0.1 (244). Phytochemical Screening Tests Identified Presence Of Alkaloids, Steroids, Terpenoids, Flavonoids, Tannins, And Saponins In The Leaves Of The Three Studied Plants. Six Pure Compounds Were Isolated Using Column Chromatography: Compound 1 From The Petroleum Ether And Chloroform (9:1) Fraction Of V. Amygdalina, And Compounds 2 -4 From O. Integrifolia And 5 And 6 From Chloroform: Methanol (1:1) Fractions Of S. Rosmarinus. The Petroleum Ether Leaf Extracts Of V. Amygdalina Exhibited Significant Antimicrobial Activity Against P. Aeruginosa, With An Inhibition Zone Of 17.80 ?? 0.2 Mm. However, Among All The Tested Extracts, The Petroleum Ether Leaf Extracts Of S. Rosmarinus Showed Strong Antimicrobial Activity, With Inhibition Zones Of 21.37 Mm Against S. Aureus And 20.83 Mm Against C. Albicans. The Study Found That The Petroleum Ether Extract Of S. Rosmarinus And The Chloroform: Methanol (1:1) Extract Of O. Integrifolia Exhibited Strong Anti-Proliferative Effects On Hela Cancer Cells. Specifically, The Anti-Proliferation Rates Were 93.73% And 91.02%, Respectively, When Tested At A Concentration Of 1250 ??G Ml?????. In The Three Plants, The Petroleum Ether Extracts From Leaves Demonstrated Higher Ic50 (??G Ml-1) Values For Dpph Radical- Scavenging Activities, Ranging From 10 To 30 ??G Ml?????, Compared To The Positive Control Ascorbic Acid. The Chloroform: Methanol (1:1) Extract Of S. Rosmarinus Leaves Exhibited The Highest Anti-Inflammatory Activity Against Albumin Denaturation, With An Ic50 Value Of 0.58 ??G Ml-1. This Was Followed By V. Amygdalina, Which Had An Ic50 Value Of 0.81 ??G Ml????? In Comparison, The Positive Control, Aspirin, Exhibited An Ic50 Value Of 4.59 ??G Ml?????. Similarly, The Extracts Of V. Amygdalina And O. Integrifolia In Chloroform/Methanol (1:1) Demonstrated Significant Inhibitory Activity Against Proteinase, With Ic50 Values Of 0.77 ??G Ml????? And 0.86 ??G Ml?????, Respectively, Both Showing Higher Activity Than The Positive Control. The Invitro Study Of Salvia Rosmarinus Extracts Chloroform: Methanol (1:1) Showed Strong Amoebicidal Activity Against Acanthamoeba Castellanii, Resulting In Less Than 10% Viability. Among All The Tested Antiviral Activities, The In Vitro Studies Of V. Amygdalina Extracts Using Aqueous, Chloroform: Methanol (1:1), And Petroleum Ether Have Shown A Selectivity Index (Si) Greater Than 10, Indicating Their Potential As Antiviral Candidates Against Influenza A (H1n1).The Essential Oils (Eos) From The Leaves Of V. Amygdalina, O. Integrifolia, And S. Rosmarinus Contained Main Components: Dihydro-2hpyran (93.6%), 3-Hexanol (19.63%), And Camphene (23.75%), Respectively. The Eos Derived From S. Rosmarinus And V. Amygdalina Exhibited The Most Potent Antifungal Properties Against C. Albicans. Among All The Tested Eos, The Eo From Salvia Rosmarinus Leaves Showed The Strongest Antioxidant Activity, As Indicated By Its Dpph Free Radical Scavenging Capacity, With An Ic50 Value Of 0.6927 ??G Ml?????. The In Silico Study Revealed That The Extracted Eos Dihydro-2h-Pyran Was Highly Bound To Two Bacterial Strains: S. Pyogenes (Pdb Id: 4d8e) With A Minimum Binding Energy Of ?��?3.3 Kcal Mol-1 And E. Coli (Pdb Id: 6f86) With ?��?2.7 Kcal Mol-1. In Comparison, Amoxicillin Had Binding Energies Of