Virginia Commonwealth University VCU Scholars Compass Theses and Dissertations Graduate School 2013 SYNTHESIS, TESTING AND CRYSTALLOGRAPHIC STUDIES OF ALLOSTERIC MODIFIERS OF HEMOGLOBIN Tanvi Deshpande Virginia Commonwealth University Follow this and additional works at: https://scholarscompass.vcu.edu/etd Part of the Pharmacy and Pharmaceutical Sciences Commons © The Author Downloaded from https://scholarscompass.vcu.edu/etd/543 This Thesis is brought to you for free and open access by the Graduate School at VCU Scholars Compass. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of VCU Scholars Compass. For more information, please contact [email protected]. © Tanvi M Deshpande 2013 All Rights Reserved SYNTHESIS, TESTING AND CRYSTALLOGRAPHIC STUDIES OF ALLOSTERIC MODIFIERS OF HEMOGLOBIN A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science at Virginia Commonwealth University. By TANVI M. DESHPANDE Bachelor of Pharmacy, Mumbai University, India, 2011 Director: DR. MARTIN SAFO ASSOCIATE PROFESSOR OF MEDICINAL CHEMISTRY Virginia Commonwealth University Richmond, Virginia July, 2013 ii ACKNOWLEDGMENTS This research project would not have been possible without the support of a number of people. I am deeply indebted to my primary advisor Dr. Martin Safo, who has been my role model in these two years of research. I would like to thank him for his constant guidance and support that instilled a whole new level of confidence in me. I would also like to thank my secondary advisor, Dr. Yan Zhang, for his supervision and advice at every stage of my research. He has been my moral support throughout the course of study. I will always be grateful to both of them for giving me the opportunity to work in their lab, and broaden my knowledge and understanding of synthetic and biological chemistry. I would like to thank Dr. Mohini Ghatge and Dr. Yunyun Yuan for their invaluable help and support. I would like to express my gratitude to both of them, Mohini for all the help during my assays and Yunyun for teaching me the basics of synthetic chemistry and also performing HPLC on my final compounds. Thank you Dr. Richmond Danso-Danquah for his chemistry guidance. Special thanks to Akul Mehta for helping me with UPLC-MS of my compounds. Many thanks to Dr. Jurgen Venitz, who in the midst of all his activities, agreed to be on my defense committee. A sincere thanks to Dr. Desai for all his advice and guidance. I would like to thank National Institute of Health, for giving me the opportunity to work in their lab. It was a wonderful experience to work in the clinical research lab. In addition, thanks to Dr. Abdulmalik for performing morphological studies on our compounds. I would like to take this opportunity to thank Sharon Lee, Michelle Wise, Jennie Kilgore, and Sharon for helping me. I would like to thank the Department of Medicinal Chemistry, Institute of Structural Biology and Drug discovery and the School of Pharmacy at Virginia Commonwealth University for giving me this opportunity to pursue my Masters. iii It was amazing to work with my group members; Christina, Courtney, Mostafa, Tom, Saheem, Chris and Dwight who made working in the lab enjoyable. My special thanks to my roommate Batul, for her words of comfort during stressful times. Thank you Aje for guiding me every step of the way. I would also like to thank Farhana, Divya, Rio, Hardik, Khushboo, Shilpa, Sweety, Soumya, Shankar, Priyanka, Reddy, Shrenik, Preetpal and Soundarya who have helped me tremendously since the day I came to Richmond. I would like to express my love to Tanushree, Kavya, Maaria, Nishant and Amey. Last but not the least, Shanty, who always stood by me, helped me through all the tough times, thank you so much! My parents and grandparents require a special acknowledgement for always being supportive of my decisions. I wouldn’t have been able to face the evils in life without their blessings on my head, especially my mom for being my source of inspiration and my anchor. I would not have been the person I am today without her presence in my life. I would also like to thank Chita, Shyam kaka, Yash and Veer for the constant support and entertainment. Finally, I would like to thank God for everything. iv TABLE OF CONTENTS List of Tables ................................................................................................................................ vii List of Figures ..............................................................................................................................viii List of Schemes ............................................................................................................................. xi List of Abbreviations ...................................................................................................................xii Abstract ........................................................................................................................................xvi I. Introduction ...............................................................................................................................1 1. Hemoglobin – A target for drug design ..............................................................................1 2. Structure and function of hemoglobin ................................................................................3 3. The oxygen equilibrium curve (OEC) ................................................................................7 4. Abnormal human hemoglobins ......................................................................................... 10 5. Sickle cell disease ............................................................................................................. 12 6. Therapeutic strategies for SCD ......................................................................................... 15 a. Natural products .......................................................................................................... 16 b. Genetic ........................................................................................................................ 16 c. Membrane acting drugs ............................................................................................... 17 d. Stereospecific acting agents ........................................................................................ 18 i. Noncovalent Hb binders ......................................................................................... 19 v ii. Covalent Hb binders ............................................................................................... 19 iii. Transient covalent binders of Hb ........................................................................... 20 e. Other agents ................................................................................................................ 22 7. Compounds that bind to Hb and decrease its oxygen affinity .......................................... 22 II. Rationale, Objectives and Specific Aims ................................................................................ 24 III. Results ................................................................................................................................... 35 IV. Discussion ............................................................................................................................. 82 V. Conclusions ........................................................................................................................... 96 VI. Experimental ......................................................................................................................... 99 1. Chemistry ........................................................................................................................99 2-Formyl-4-methoxyphenyl isonicotinate (35) ..............................................................100 N-(2-Formyl-4-methoxyphenyl) isonicotinamide (36) ..................................................101 2-((6-(Hydroxymethyl)pyridin-2-yl)methoxy)-5-methoxybenzaldehyde (37) ..............102 2-((6-(Hydroxymethyl)pyridin-2-yl)methoxy)-4-methoxybenzaldehyde (38) ..............103 3-((6-(Hydroxymethyl)pyridine-2-yl)methoxy)-4-methoxybenzaldehyde (39) .............104 5-Nitrooxymethyl furfural (40) ......................................................................................105 (6-((2-Formyl-4-methoxyphenoxy)methyl)pyridin-2-yl)methyl nitrate (41) .................106 (6-((2-Formyl-5-methoxyphenoxy)methyl)pyridin-2-yl)methyl nitrate (42) .................107 (6-((5-Formyl-2-methoxyphenoxy)methyl)pyridin-2-yl)methyl nitrate (43) .................108 2-(Nitrooxy)ethyl 2-(4-(2-((3,5-dimethylphenyl)amino)-2-oxoethyl)phenoxy)-2- ethylpropanoate (44) .......................................................................................................109 5-Methoxy-2-nitrobenzaldehyde (49) ............................................................................110 2-Amino-5-methoxybenzaldehyde (50) .........................................................................110 vi 2-((6-(Bromomethyl)pyridin-2-yl)methoxy)-5-methoxybenzaldehyde (54) .................111 2-((6-(Bromomethyl)pyridin-2-yl)methoxy)-4-methoxybenzaldehyde (55) .................112 3-((6-(Bromomethyl)pyridine-2-yl)methoxy)-4-methoxybenzaldehyde (56) ................113 2-Bromoethyl nitrate (58) ...............................................................................................114