Infectious diseases are the second leading cause of deaths in the world with malaria being responsible for approximately the same amount of deaths as cancer in 2012. Despite the success in malaria prevention and control measures decreasing the disease mortality rate by 45% since 2000, the development of single-dose therapeutics with radical cure potential is required to completely eradicate this deadly disease. Targeting multiple stages of the malaria parasite is becoming a primary requirement for new candidates in antimalarial drug discovery and development. Recently, 4(1H)-pyridone, 4(1H)-quinolone, 1,2,3,4-tetrahydroacridone, and phenoxyethoxy-4(1H)-quinolone chemotypes have been shown to be antimalarials with blood stage activity, liver stage activity, and transmission blocking activity. Advancements in structure-activity relationship and structure-property relationship studies, biological evaluation in vitro and in vivo, as well as pharmacokinetics of the 4(1H)-pyridone and 4(1H)-quinolone chemotypes is discussed in the first chapter of the dissertation. Convenient synthetic approaches to 3-aryl-4(1H)-quinolones via metal-catalyzed and metal-free arylation of β-keto carbonyl compounds is addressed in Chapter 2. A clean arylation protocol of ethyl acetoacetate was developed by using hypervalent diaryl iodonium salts under mild and metal-free conditions. The scope of the reaction, using symmetric and unsymmetric iodonium salts varying in sterics and electronics was examined. This method has been applied for the synthesis of antimalarial compound ELQ-300, which is currently in preclinical development. Additionally, a first gram scale synthesis of ELQ-300 and its structurally related 4(1H)-quinolone P4Q-391 using operationally simple and highly yielding metal-catalyzed conditions have been shown. Despite of 3-aryl-4(1H)-quinolone chemotypes displaying potent antimalarial activities against Plasmodium species in vitro and in vivo, their development is also associated with risks. 4(1H)-quinolones are known to be poorly soluble and thus represent challenging drug candidates for pharmacokinetic and bioavailability reasons. Disrupting of molecular crystal packing and prodrug approaches were employed to overcome solubility and bioavailability issues in current series. Quantum mechanics torsion profile calculations, 13C T1 spin-lattice relaxation experiments as well as X-ray studies were conducted with the objective to determine possible effects improving key physicochemical properties such as solubility and stability. As a backup strategy, a prodrug approach was developed enabling the 4(1H)-quinolone scaffold to be functionalized at the quinolone's oxygen. In order to avoid any enzymatic dependences, an approach was developed in which the prodrug moiety was removed via a pH-triggered decay. Additionally, phosphate prodrugs regenerating the active compound via extrahepatic enzymes such as the ubiquitous alkaline phosphatase were investigated. The development of orally bioavailable prodrugs enabled an advance overcoming in vivo efficacy limitations and has been confirmed by pharmacokinetic profiling studies. The herein presented approaches present viable options for any pyridone quinolone antimalarial chemotype which are currently studied.

The goal of our research endeavor was to successfully employ modern lead discovery and optimization strategies towards the development and identification of compounds possessing antimalarial activity. Preliminary data from in vitro screening at the Walter Reed Army Institute of Research identified several chemotypes including 4(1H)-quinolones and 1,2,3,4-tetrahydroacridones to have potent antimalarial activities. Multiple synthetic routes were devised and implemented which enabled the rapid preparation and isolation of over 400 structurally diverse 4(1H)-quinolones and 1,2,3,4-tetrahydroacridones. Our research towards discovering and optimizing antimalarials was inspired from the severe impact malaria has had on our planet especially on impoverished countries. There are over 300 million cases annually and over one million deaths. The staggering mortality rates combined with the global emergence of chemical resistance that the parasite Plasmodium falciparum has developed towards many of the common antimalarials compelled us to extend our research efforts to this growing problem. The need for identifying and developing new antimalarial drugs is very important. However, our approach focuses on the optimization of historic antimalarials such as endochin, floxacrine, or ICI 56,780 which possess liabilities such as lack of poor solubility, poor in vivo activity or lingering toxicity issues. Through these optimization efforts using both SAR and structure-property relationship (SPR) studies, a more suitable candidate was developed that had superior physicochemical properties. Our drug design approach included not only the identification of liabilities of historic compounds but also the synthesis and optimization of numerous analogs guided by SAR. All compounds were tested in vitro for antimalarial activity and characterized in parallel for physicochemical properties such as solubility, permeability, and logD7.4. Insights from both the antimalarial activity as well as the physicochemical properties determined which analogs would be advanced in the design process. Based on our early investigations, 6-chloro-7-methoxy-3-phenyl-4(1H)-quinolone emerged as a promising hit. Compared to endochin, which possesses EC50s of 8.6 nM and 46.6 nM against drug resistant strains W2 and TM90C2B, and a solubility of less than 2 µM, 6-chloro-7-methoxy-3-pheny-4(1H)-quinolone was superior with a 4-fold improvement in solubility (6 µM) as well as slightly improved antimalarial activity (EC50s of 26.2 nM and 15.3 nM against W2 and TM90C2B, respectively). Unfortunately, this compound failed to reduce parasitemia levels in P. berghei infected mice. Hit-to-lead optimization lead to the discovery of 6-chloro-7-methoxy-2-methyl-3-o-tolyl-4(1H)-quinolone which was shown to reduce parasitemia levels by 41% at day 6 post-exposure (PE) in P. berghei infected mice at a 50 mg/kg dose. The observed in vivo activity of 6-chloro-7-methoxy-2-methyl-3-o-tolyl-4(1H)-quinolone was believed to relate to the 3-fold increase in solubility (19 µM) over the 3-phenyl-susbtituted analogue. Continuation of SAR and SPR studies identified additional 4(1H)-quinolones suggesting that the microsomal stability of the compounds is as important for in vivo efficacy as the aqueous solubility. Several of the analogs that showed minimal degradation in human microsomal stability studies demonstrated increased in vivo activity in the ranges of 72-98% parasitemia reductions on day 6PE in P. berghei infected mice at 50 mg/kg. These results helped refine the final SAR and SPR optimization identifying a compound with radical curative activity in mice (99% parasitemia reductions on day 6PE in P. berghei infected mice at 50 mg/kg with five out of five mice surviving beyond 30 days). Theses studies not only highlight the effectiveness of detailed SAR and SPR strategies used in drug discovery programs, but they also showcase the importance of re-evaluating historic antimalarials and exploiting their shortcomings. These studies have opened the doors to several possibilities regarding the 4(1H)-quinolone scaffold optimization for future antimalarial development. Several of the compounds described in this work are currently being subjected to stringent head-to-head comparative studies to determine the analog best suited for pre-clinical trials.