Phytochemical Analysis, Biological Activities, And Computational Study Of Compounds Isolated From Selected Medicinal Plants Of Ethiopian Flora

Abstract

Drug Resistance, The Side Effects And High Cost Associated With The Existing Antibiotics, Reappearance Of Previously Known Diseases, And Changes In Genetic Make-Up Are Major Problems In The Treatment Of Human Health. Thus, The Search For Alternatively Safe And Effective Drugs With Antimicrobial And Anticancer Agent Has Become A Necessity. Plants Have Been Known As Potential Source Of Therapeutic Agents For The Treatment Of Various Diseases Since The Ancient Days. Nearly 80% Of The Population In Ethiopia Uses Plant-Based Traditional Medicine As Their Primary Health Care. In The Traditional Medicine Of Ethiopia, Cyphostema Cyphopetalum, Hydnora Johannis, Capparis Tomentosa, Kalanchoe Petitiana And Cadaba Rotundifolia Are Used As Medicinal Plants For The Treatment Of Various Ailments. The Objective Of This Study Was To Analyse The Phytochemical Constituents, Evaluate Biological Activities Of Extracts And Isolated Compounds, And Predict Computational Study Of Compounds Isolated From Roots Of Cyphostemmacyphoptealum And Hydrona Johannis, Stem Bark Of Capparis Tomentosa, And Leaves Of Kalanchoe Petitena And C.Adaba Rotundifolia. Extraction By Maceration Was Used To Obtain Crude Extracts. Silica Gel Column Chromatography Was Used To Isolate Compounds. Spectroscopic Techniques (Ir, 1d (1h, 13c, And Dept -135) And 2d (Cosy, Hsqc, And Hmbc Nmr) Were Used To Elucidate Structures Of Isolated Compounds. Hydrodistialatin And Gc-Mswere Used To Extract And Analyse The Chemical Composition Of Essential Oils, Respectively. Disc Diffusion Assay Was Used For In Vitro Antibacterial And Antifungal Activity Evaluation Of Isolated Compounds And Extracts Against Bacterial Strains Escherichia Coli, Pseudomonas Aeruginosa, Staphylococcus Aureus And Streptococcus Pyogen, And Fungal Strain Candida Albicans. The Dpph Assay Was Used To Evaluate Antioxidant Activity Of Isolated Compounds. Mtt Assay Was Used To Evaluate The Cytotoxicity Of Extracts From Roots Of C. Cyphopetalum, And H. Johannis, And Stem Bark Of C. Tomentosa Against Mcf-7 Cell Lines. Autodock Vina 4.2 (Mgl Tools 1.5.7) Was Used To Perform The Molecular Docking Analysis To Predict The Potential Binding Modes Of The Isolated Compounds With The Target Bacterial Proteins (Dna Gyraseb, S. Aureus Pk, Pqsa). Swissadme And Preadmet Tools Were Used To Predict The In Silico Drug Likeness And Admeproperties Of Isolated Compounds. Protox Ii Tool To Predict Insilioc Toxicity. Dft Analysis Of Isolated Compounds Was Performed Using Gaussian 09 And Visualized Through Gauss View 6.0.Silica Gel Column Chromatographic Separation Of Extracts Of Five Selected Medicinal Plants Afforded 20 Compounds, Including A New Hydroxyl-Spongiane Diterpenoid Lactone Derivative, 3-Hydroxyisoagatholactone(100) Along With Trans-Resveratrol (28), ??-Sitosterol (49) And ?? Viniferin (101) From Dichloromethane/Methanol (1:1), And Gnetin H (102), Tricuspidatol A (103), ??-Viniferin-Diol (104) And Parthenostilbenin B (105) From Methanol Root Extract Of C. Cyphopetalum. Silmilarily, Catechin (40), Oleic Acid (41), ??-Sitosterol (49), Catechin-7-O Glucoside (106), Sitosterol-3-O-Glucoside (107), Caffeic Acid Derivative (108), And A Pregn Derivative (3,9-Epoxypregn-16-Ene-14,20-Diol,7,11,18-Triacetoxy-3-Methox) (109) Were Isolated From Roots Of H. Johannis. The Dichloromethane/Methaol Extract Of Stem Barck Of C. Tomentosa Afforded Tetradecanoic Acid (110) And 4-Hyroxybenzoate Derivative (111), Leaf Extract Of K. Petitiana And C. Rotundifolia Afforded Bis-(2-Ethylhexyl) Phthalate (112) And Diethyl Phthalate (113). The Gc-Ms Analysis Of Essential Oil Obtained From Leaves Of C.Tomentosa, K. Petitiana And C. Rotundifolia Resulted In Identification Phenol, 2,4-Bis(1,1-Dimethylethyl)-(21.03 %), 1,5-Hexadiene, 2-Methyl (23.07 %), Phenol, 2,4-Bis(1,1-Dimethylethyl)- (15.6%) As Major Compounds, Respectively. Among The Compounds Isolated From Cyphostema Cyphopetalum, At 50 ??G/Ml, The Highest Growth Inhibition Against E. Coli Was 2exhibited By ??-Viniferin (101) (8.55 ?? 0.45 Mm) And Tricuspidatol A (103) (8.42??0.20 Mm). Whereas, Against P. Aeruginosa And S. Aureus The Highest Inhibition Zone Was Scored By Gnetin H (102) (12.55 ?? 0.65 Mm) And ??-Viniferin (101) (9.30??1.39 Mm) Respectively, Which Is Good Compared To Chloramphenicol (19.06 ?? 2.28, 11.76 ?? 0.77, And 15.74 ?? 1.02, At 30 ??G/Ml, Respectively). The Dcm/Meoh Extract Of Roots Of H. Johannis Showed The Highest Antibacterialactivity Against S. Aureus (10.75??0.25 Mm), P. Aeruginosa (10.5??0.5 Mm), And E. Coli (9.5 ?? 0.5 Mm), And N-Hexane Against S. Pyogenes (9.5 ?? 0.5 Mm). Strong Antibacterial Activities Were Exhibited By Compound 108 Against P. Aeruginosa (13.72??0.05 Mm, 0.25 Mg/Ml) And Compound 107 Against S. Aureus (10.75 ?? 0.25 Mm) At 0.25 Mg/Ml, Compared To Amoxicillin (16.00, 15.25,Respectively At 0.25 Mg/Ml). The Dcm/Meoh (1:1) Extract Of Roots Of C. Cyphopetalum Showed Moderate Antifungal Activity (8.5??0.5 Mm, At 12.5 Mg/Ml) Compared To Ketoconazole (17.3??2.05 Mm, At 10 ??G/Disc). The Methanol Extracts Of The Roots Of C. Cyphopetalum And H. Johannis, And Dichloromethane/Methanol (1:1) Extract Of Stem Bark Extract Of C. Tomentosa Showed Strong Anticancer Activity Against Breast Cancer Cell Line (Mcf-7) At % Cell Viability<50% Each. At Concentration Of 12.5 ??G/Ml, The Highest Percentage Inhibition Was Recorded By Compound 28 (76.87??0.09 %), Followed By Compound 101 (74.30??0.12 %), And 105 (73.32??0.25 %), With Ic50 Value Of 0.052 ??G/Ml, 0.017 ??G/Ml, And 0.025??G/Ml, Respectively.The Molecular Docking Analysis Revealed That, Highest Binding Affinity Toward Dna Gyrase Was Exhibited By Compounds 101 And 107 (-7.6 Kcal/Mol). Compounds 102, 107 And 40 (-8.8, -8.7, And -8.6, Respectively) Displayed The Next Highest Binding Affinity Towards Protein Pqsa Whereas Compound 107 (-7.4 Kcal/Mol) Showed Promising Binding Affinity Towards S. Aureus Pkcompared To Chloramphenicol (-6.4, -7.0, And -4.6 Kcal/Mol, Respectively). Compounds 28, 100, And 101 Satisfy Lipinski?�?S Rule Of Five With Zero Violations. Compounds 40 And 107 Exhibited Negligible Acute Toxicity (Ld50>5000, Toxicity Class>5). Compound 28 Recorded The Lowest Homo-Lumo Energy Gap And Good Chemical Activity. In This Work, The Computational Study Results Are In Agreement With The In Vitro Experimental Studies Obtained For Compounds. Thus,Compounds 28, 100, 101, 102, 106 And 107 May Have Potential To Be Used As Lead Molecules That Could Be Developed Into Potent Bacterial Growth Inhibitors. This Supported The Traditional Uses Exhibited By ??-Viniferin (101) (8.55 ?? 0.45 Mm) And Tricuspidatol A (103) (8.42??0.20 Mm). Whereas, Against P. Aeruginosa And S. Aureus The Highest Inhibition Zone Was Scored By Gnetin H (102) (12.55 ?? 0.65 Mm) And ??-Viniferin (101) (9.30??1.39 Mm) Respectively, Which Is Good Compared To Chloramphenicol (19.06 ?? 2.28, 11.76 ?? 0.77, And 15.74 ?? 1.02, At 30 ??G/Ml, Respectively). The Dcm/Meoh Extract Of Roots Of H. Johannis Showed The Highest Antibacterialactivity Against S. Aureus (10.75??0.25 Mm), P. Aeruginosa (10.5??0.5 Mm), And E. Coli (9.5 ?? 0.5 Mm), And N-Hexane Against S. Pyogenes (9.5 ?? 0.5 Mm). Strong Antibacterial Activities Were Exhibited By Compound 108 Against P. Aeruginosa (13.72??0.05 Mm, 0.25 Mg/Ml) And Compound 107 Against S. Aureus (10.75 ?? 0.25 Mm) At 0.25 Mg/Ml, Compared To Amoxicillin (16.00, 15.25,Respectively At 0.25 Mg/Ml). The Dcm/Meoh (1:1) Extract Of Roots Of C. Cyphopetalum Showed Moderate Antifungal Activity (8.5??0.5 Mm, At 12.5 Mg/Ml) Compared To Ketoconazole (17.3??2.05 Mm, At 10 ??G/Disc). The Methanol Extracts Of The Roots Of C. Cyphopetalum And H. Johannis, And Dichloromethane/Methanol (1:1) Extract Of Stem Bark Extract Of C. Tomentosa Showed Strong Anticancer Activity Against Breast Cancer Cell Line (Mcf-7) At % Cell Viability<50% Each. At Concentration Of 12.5 ??G/Ml, The Highest Percentage Inhibition Was Recorded By Compound 28 (76.87??0.09 %), Followed By Compound 101 (74.30??0.12 %), And 105 (73.32??0.25 %), With Ic50 Value Of 0.052 ??G/Ml, 0.017 ??G/Ml, And 0.025??G/Ml, Respectively.The Molecular Docking Analysis Revealed That, Highest Binding Affinity Toward Dna Gyrase Was Exhibited By Compounds 101 And 107 (-7.6 Kcal/Mol). Compounds 102, 107 And 40 (-8.8, -8.7, And -8.6, Respectively) Displayed The Next Highest Binding Affinity Towards Protein Pqsa Whereas Compound 107 (-7.4 Kcal/Mol) Showed Promising Binding Affinity Towards S. Aureus Pkcompared To Chloramphenicol (-6.4, -7.0, And -4.6 Kcal/Mol, Respectively). Compounds 28, 100, Respectively) Displayed The Next Highest Binding Affinity Towards Protein Pqsa Whereas Compound 107 (-7.4 Kcal/Mol) Showed Promising Binding Affinity Towards S. Aureus Pkcompared To Chloramphenicol (-6.4, -7.0, And -4.6 Kcal/Mol, Respectively). Compounds 28, 100, And 101 Satisfy Lipinski?��?S Rule Of Five With Zero Violations. Compounds 40 And 107 Exhibited Negligible Acute Toxicity (Ld50>5000, Toxicity Class>5). Compound 28 Recorded The Lowest Homo-Lumo Energy Gap And Good Chemical Activity. In This Work, The Computational Study Results Are In Agreement With The In Vitro Experimental Studies Obtained For Compounds. Thus,Compounds 28, 100, 101, 102, 106 And 107 May Have Potential To Be