Bismillah Hirrahman Nirraheem In the Name of Allah, the Most Gracious, the Most Merciful All praises for Almighty Allah, the most kind and cordial. He is ample and the best disposer of all affairs (for us). I am showing my humble submission to the heart and soul after The Holy Prophet Hazrat Muhammad (sallallahualayhewassallum) whose life is an ideal pattern for all of us . Phytochemical Studies on the Chemical Constituents of Xanthium strumarium Linn., Synthesis in addition Bioactivities of 2, 3-Diaminonaphthalenimidazole Derivatives and Amides of Piperic Acid Thesis submitted for The partial fulfillment of the Degree of DOCTOR OF PHILOSOPHY in Chemistry by AMINA SULTANA Department of Chemistry Federal Urdu University of Arts, Science and Technology Karachi – 75260, Pakistan 2014 CERTIFICATE Certified that Ms. Amina Sultana has carried out her research work on the topic entitled Phytochemical Studies on the Chemical Constituents of Xanthium strumarium Linn., Synthesis in addition Bioactivities of 2, 3- Diaminonaphtalenimidazole Derivatives and Amides of Piperic Acid at Department of Chemistry, Federal Urdu University of Arts, Science, and Technology, Karachi, Pakistan, under the supervision of Dr. Aneela Wahab. Her research work is original that has not been submitted to any other university. Some publications have been earned from the course of this research work and the all are acknowledged. Dr. Aneela Wahab --------------------------- Assistant Professor Department of Chemistry Federal Urdu University, Karachi, Pakistan Research Supervisor Dr. Iffat Mahmood --------------------------- Chairperson Department of Chemistry Federal Urdu University, Karachi, Pakistan Dedicated to my loving and caring Parents whose prayers have always been a great source of Strength to me Contents Acknowledgement --------------------------------------------------------------------- 1 Summary --------------------------------------------------------------------------------- 3 KHULASA ------------------------------------------------------------------------------- 6 CHAPTER -1 Phytochemical Studies on the Chemical Constituents of Xanthium strumarium Linn. ----------------------------------------------------- 10 1.0 Introduction -------------------------------------------------------------------------- 11 1.1. General Introduction --------------------------------------------------------------- 12 1.1.1. Classification ------------------------------------------------------------ 17 1.1.2. Introduction About Family Compositeae ---------------------------- 18 1.1.3. Introduction About Genus Xanthium --------------------------------- 20 1.1.4. General Description of Xanthium strumarium Linn. --------------- 21 1.1.4.1. Chemical Constituents of Xanthium strumarium Linn.---------- 21 1.1.4.2. Medicinal Importance of Xanthium strumarium Linn.----------- 22 1.1.4.3. Literature Review ----------------------------------------------------- 24 1.2. Present Work ----------------------------------------------------------------------- 53 1.3. Results and Discussion ------------------------------------------------------------ 55 1.3.1. Lupenyl acetate (1) ----------------------------------------------------- 56 1.3.2. Stigmasterol (2) --------------------------------------------------------- 59 1.3.3. β-Sitosterol (3) ------------------------------------------------------- 60 1.3.4. Palmitic acid (4) ---------------------------------------------------------61 1.3.5. β-Amyrin (5) ------------------------------------------------------------ 62 1.3.6. Oleanolic acid (6) ------------------------------------------------------- 63 1.3.7. β-Sitosterol-3-O-β-D-Glucopyranoside (7) ------------------------- 64 1.3.8. Ferulic acid (8) ---------------------------------------------------------- 65 1.3.9. Biological Activities ---------------------------------------------------- 66 1.3.9.1. In vitro Anti-bacterial Activity ------------------------------------- 66 1.3.9.2. In vitro Anti-fungal Activity ---------------------------------------- 68 1.3.9.3. In vitro Anti-oxidant Activity --------------------------------------- 70 1.4. Experimental ------------------------------------------------------------------------ 71 1.4.1. General Experimental -------------------------------------------------- 72 1.4.2. Collection of Plant Material ------------------------------------------- 72 1.4.3. Extraction and Isolation ------------------------------------------------ 72 1.4.4. Characterization of Compounds -------------------------------------- 79 1.4.4.1. Characterization of Lupenyl acetate (1) -------------------------- 79 1.4.4.2. Characterization of Stigmasterol (2) ------------------------------- 80 1.4.4.3. Characterization of β-Sitosterol (3) -------------------------------- 81 1.4.4.4. Characterization of Palmitic acid (4) ------------------------------ 82 1.4.4.5. Characterization of β-Amyrin (5) ---------------------------------- 83 1.4.4.6. Characterization of Oleanolic acid (6) ---------------------------- 84 1.4.4.7. Characterization of β-Sitosterol-3-O-β-D- Glucopyranoside (7)-------------------------------------------------- 85 1.4.4.8. Characterization of Ferulic acid (8) ------------------------------- 86 1.5. References ------------------------------------------------------------------------- 87 CHAPTER-2 Synthesis in addition Bioactivities of 2, 3-Diaminonaphthalenimidazole Derivatives----------------------------------------------------------------------------- 97 2.0. General Introduction For Chapter-2 and 3 ----------------------------------- 99 2.1. Introduction of Benzimidazole -------------------------------------------------- 104 2.1.1 Biological Importance ----------------------------------------------------- 104 2.2. Synthetic Approaches Towards Benzimidazole ------------------------------ 112 2.3. Results and Discussion ----------------------------------------------------------- 123 2.3.1. Chemistry ------------------------------------------------------------------- 124 2.3.2. General Method for the Synthesis of Compounds (65-99) ---------- 124 2.3.3. General Stucture Elucidation --------------------------------------------- 129 2.3.4. Biological Evaluation of Compounds (65-99) ------------------------- 131 2.3.4.1. In Vitro Tyrosinase Inhibitory Activity ----------------------------- 131 2.3.4.2. In Vitro Acetylcholinesterase and Butrylcholinesterase. Inhibitory Activity ------------------------------------------------------ 133 2.3.4.3. In Vitro Urease Inhibitory Activity -------------------------------- 136 2.3.4.4. In Vitro Anti-bacterial Activity ------------------------------------- 138 2.3.4.5. In Vitro Anti-fungal Activity ---------------------------------------- 146 2.3.4.6. In Vitro Anti-oxidant Activity -------------------------------------- 149 2.3.5. Conclusion ------------------------------------------------------------------- 151 2.4. Experimental ------------------------------------------------------------------------ 152 2.4.1. General Experimental ------------------------------------------------------ 153 2.4.2. General Method for the Synthesis of Compounds (65-99) ----------- 153 2.4.2.1. 2-(1H-indol-3-yl)-1H-naphtho[2,3-d]imidazole (65) --------------- 154 2.4.2.2. 2-(4-ethoxy-3-methoxyphenyl)-1H-naphtho[2,3-d]imidazole (66)-154 2.4.2.3. 4-(1H-naphtho[2,3-d]imidazol-2-yl)phenol (67) -------------------- 154 2.4.2.4. 2-methoxy-4-(1H-naphtho[2,3-d]imidazol-2-yl) phenyl acetate (68) -------------------------------------------------------- 154 2.4.2.5. 2-(5-bromo-2-methoxyphenyl)-1H-naphtho[2,3-d]imidazole (69)- 155 2.4.2.6. 2-(2, 5-dimethoxyphenyl)-1H-naphtho[2,3-d]imidazole (70) ----- 155 2.4.2.7. 4-chloro-2-(1H-naphtho[2,3-d]imidazol-2-yl)phenol (71) --------- 156 2.4.2.8. 2-(4-trifluoromethylphenyl)-1H-naphtho[2,3-d]imidazole (72) --- 156 2.4.2.9. 2-(4-nitrophenyl)-1H-naphtho[2,3-d]imidazole (73) --------------- 156 2.4.2.10. 2, 6-dimethoxy-4-(1H-naphtho[2,3-d]imidazol-2-yl)phenol (74) -157 2.4.2.11. 2-(3-benzyloxyphenyl)-1H-naphtho[2,3-d]imidazole (75) -------- 157 2.4.2.12. 2-(2-fluoro-4-methoxyphenyl)-1H-naphtho[2,3-d] imidazole (76)------------------------------------------------------------ 157 2.4.2.13. 2-methoxy-5-(1H-naphtho[2,3-d]imidazol-2-yl)phenol (77) ----- 157 2.4.2.14. 2-(4-benzyloxyphenyl)-1H-naphtho[2,3-d]imidazole (78) ------- 158 2.4.2.15. 2-(3-ethoxy-4-methoxyphenyl)-1H-naphtho[2,3-d] imidazole (79)------------------------------------------------------------ 158 2.4.2.16. 4-(1H-naphtho[2,3-d]imidazol-2-yl)-3-nitrophenol (80) ---------- 158 2.4.2.17. 2-(thiophen-2-yl)-1H-naphtho[2,3-d]imidazole (81) --------------- 159 2.4.2.18. 2-(3, 4-dimethoxyphenyl)-1H-naphtho[2,3-d]imidazole (82) ---- 159 2.4.2.19. 4-(1H-naphtho[2,3-d]imidazol-2-yl)benzene-1,3-diol (83) ------- 159 2.4.2.20. 2-(2, 3, 4-trimethoxyphenyl)-1H-naphtho[2,3-d]imidazole (84)-- 160 2.4.2.21. 4-(1H-naphtho[2,3-d]imidazol-2-yl)benzene-1,2,3-triol (85) ----- 160 2.4.2.22. 2-(4-methylthiophenyl)-1H-naphtho[2,3-d]imidazole (86) ------- 160 2.4.2.23. 2-(2-nitrophenyl)-1H-naphtho[2,3-d]imidazole (87) -------------- 161 2.4.2.24. 2-(naphthalen-2-yl)-1H-naphtho[2,3-d]imidazole (88) ------------ 161 2.4.2.25. N, N-dimethyl-4-(1H-naphtho[2,3-d]imidazol-2-yl)aniline (89)-- 161 2.4.2.26. 3-bromo-6-methoxy-2-(1H-naphtho[2,3-d] imidazol-2-yl) phenol (90) --------------------------------------------------------------- 162 2.4.2.27. 2-(2-bromo-4, 5-dimethoxyphenyl)-1H-naphtho [2,3-d] imidazole (91) ----------------------------------------------------------- 162 2.4.2.28.