Novel inhibitors of phosphate-binding enzymes as potential human therapeutics

Studies on the discovery and development of small-molecule enzyme inhibitors are continuously growing due to their application as drugs for essentially every type of human disease, including cancer, metabolic, cardiovascular, neurodegenerative, and infectious diseases. However, the design of high affinity and membrane permeable inhibitors for phosphate- or pyrophosphate- binding enzymes with highly charged and metal-dependent active site pockets, such as the HIV-1 reverse transcriptase (HIV-1 RT) and the human geranylgeranyl pyrophosphate synthase (hGGPPS), poses a significant challenge in medicinal chemistry. In the past, the problems associated with this class of biological targets have been overcome with the development of effective phosphate or pyrophosphate bioisosteres and prodrugs or through the discovery of allosteric inhibitors.Guided by several successful incorporations of phosphate and pyrophosphate mimics, our studies were dedicated to the following: (1) identification and development of bona fide active site inhibitors of HIV-1 RT with a mechanism of action that is uniquely different from the currently known anti-HIV/AIDS drugs, and (2) discovery of potent and selective inhibitors of hGGPPS that can be used as molecular probes to investigate the role of hGGPPS in human diseases. The design, synthesis, and preliminary biological profiling of these novel compounds will be discussed.Les études portant sur la découverte et le développement de petites molécules jouant le rôle d'inhibiteurs enzymatiques ne cessent de s'accroître en raison de leurs applications en tant que médicaments pour un large spectre de maladies humaines telles que le cancer ou encore les maladies métaboliques, cardiovasculaires, neurodégénératives et infectieuses. Cependant, la conception d'inhibiteurs enzymatiques perméables aux membranes cellulaires ayant une grande affinité pour le site actif, qui lui-même est dépendant d'un métal hautement chargé qui est normalement lié à des substrats comportant des groupements phosphates et pyrophosphates, représente un défi très important en chimie médicinale. Nous nous sommes particulièrement intéressés aux inhibiteurs de la transcriptase inverse du VIH-1 (VIH-1 RT) et à la géranylgéranyl pyrophosphate synthétase humaine (GGPPSh). Dans le passé, les problèmes associés à cette classe de cibles biologiques ont pu être surmontés soit avec le développement de bioisostères de type phosphate ou pyrophosphate, de promédicaments efficaces ou par la découverte d'inhibiteurs allostériques.Guidés par plusieurs implémentations à nos inhibiteurs de groupements imitant les groupes fonctionnels phosphates et pyrophosphates, nos études ont été consacrées à: (1) l'identification et le développement d'inhibiteurs du site actif du VIH-1 RT bona fide avec un mécanisme d'action unique en comparaison aux médicaments actuels prescrits pour le traitement VIH/SIDA; et, (2) la découverte d'inhibiteurs puissants et sélectifs de GGPPSh qui peuvent être utilisés comme sondes moléculaires pour étudier les différents rôles de l'enzyme GGPPSh dans le traitement des maladies humaines. La conception ainsi que la synthèse et le profilage biologique préliminaire de ces nouveaux composés seront discutés dans ce manuscrit de thèse de doctorat

Novel inhibitors of phosphate-binding enzymes as potential human therapeutics

Studies on the discovery and development of small-molecule enzyme inhibitors are continuously growing due to their application as drugs for essentially every type of human disease, including cancer, metabolic, cardiovascular, neurodegenerative, and infectious diseases. However, the design of high affinity and membrane permeable inhibitors for phosphate- or pyrophosphate- binding enzymes with highly charged and metal-dependent active site pockets, such as the HIV-1 reverse transcriptase (HIV-1 RT) and the human geranylgeranyl pyrophosphate synthase (hGGPPS), poses a significant challenge in medicinal chemistry. In the past, the problems associated with this class of biological targets have been overcome with the development of effective phosphate or pyrophosphate bioisosteres and prodrugs or through the discovery of allosteric inhibitors.Guided by several successful incorporations of phosphate and pyrophosphate mimics, our studies were dedicated to the following: (1) identification and development of bona fide active site inhibitors of HIV-1 RT with a mechanism of action that is uniquely different from the currently known anti-HIV/AIDS drugs, and (2) discovery of potent and selective inhibitors of hGGPPS that can be used as molecular probes to investigate the role of hGGPPS in human diseases. The design, synthesis, and preliminary biological profiling of these novel compounds will be discussed.Les études portant sur la découverte et le développement de petites molécules jouant le rôle d'inhibiteurs enzymatiques ne cessent de s'accroître en raison de leurs applications en tant que médicaments pour un large spectre de maladies humaines telles que le cancer ou encore les maladies métaboliques, cardiovasculaires, neurodégénératives et infectieuses. Cependant, la conception d'inhibiteurs enzymatiques perméables aux membranes cellulaires ayant une grande affinité pour le site actif, qui lui-même est dépendant d'un métal hautement chargé qui est normalement lié à des substrats comportant des groupements phosphates et pyrophosphates, représente un défi très important en chimie médicinale. Nous nous sommes particulièrement intéressés aux inhibiteurs de la transcriptase inverse du VIH-1 (VIH-1 RT) et à la géranylgéranyl pyrophosphate synthétase humaine (GGPPSh). Dans le passé, les problèmes associés à cette classe de cibles biologiques ont pu être surmontés soit avec le développement de bioisostères de type phosphate ou pyrophosphate, de promédicaments efficaces ou par la découverte d'inhibiteurs allostériques.Guidés par plusieurs implémentations à nos inhibiteurs de groupements imitant les groupes fonctionnels phosphates et pyrophosphates, nos études ont été consacrées à: (1) l'identification et le développement d'inhibiteurs du site actif du VIH-1 RT bona fide avec un mécanisme d'action unique en comparaison aux médicaments actuels prescrits pour le traitement VIH/SIDA; et, (2) la découverte d'inhibiteurs puissants et sélectifs de GGPPSh qui peuvent être utilisés comme sondes moléculaires pour étudier les différents rôles de l'enzyme GGPPSh dans le traitement des maladies humaines. La conception ainsi que la synthèse et le profilage biologique préliminaire de ces nouveaux composés seront discutés dans ce manuscrit de thèse de doctorat

Modular Assembly of Purine-like Bisphosphonates as Inhibitors of HIV‑1 Reverse Transcriptase

Bisphosphonates can mimic the pyrophosphate leaving group of the nucleotidyl transfer reaction and effectively inhibit RNA/DNA polymerases. In a search of HIV-1 reverse transcriptase (RT) inhibitors, a new chemotype of nonhydrolyzable purine diphosphate mimic was synthesized. A modular synthetic protocol was developed, utilizing 2-amino-6-(methylthio)-4-(trimethylsilyl)­nicotinonitrile as the key synthon in the preparation of highly substituted 2-aminonicotinonitriles. These building blocks were subsequently elaborated to the pyrido­[2,3-<i>d</i>]­pyrimidine bisphosphonates (PYPY-BPs). Biochemical screening identified analogs of PYPY-BPs that inhibit HIV-1 RT-catalyzed DNA synthesis

A One-Step, Atom Economical Synthesis of Thieno[2,3- d

A Na2HPO4-catalyzed four-component reaction between a ketone, malononitrile, S-8 and formamide has been realized for the first time. This reaction provides a concise approach to thieno[2,3-d]pyrimidin-4-amines, previously requiring 5 steps. The utility of this reaction was validated by preparing a multi-targeted kinase inhibitor and an inhibitor of the NRF2 pathway with excellent atom- and step-economy.NIH [ES023758]12 month embargo; first published: 30 April 2019This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Pharmacophore Mapping of Thienopyrimidine-Based Monophosphonate (ThP-MP) Inhibitors of the Human Farnesyl Pyrophosphate Synthase

The human farnesyl pyrophosphate synthase (hFPPS), a key regulatory enzyme in the mevalonate pathway, catalyzes the biosynthesis of the C-15 isoprenoid farnesyl pyrophosphate (FPP). FPP plays a crucial role in the post-translational prenylation of small GTPases that perform a plethora of cellular functions. Although hFPPS is a well-established therapeutic target for lytic bone diseases, the currently available bisphosphonate drugs exhibit poor cellular uptake and distribution into nonskeletal tissues. Recent drug discovery efforts have focused primarily on allosteric inhibition of hFPPS and the discovery of non-bisphosphonate drugs for potentially treating nonskeletal diseases. Hit-to-lead optimization of a new series of thienopyrimidine-based monosphosphonates (ThP-MPs) led to the identification of analogs with nanomolar potency in inhibiting hFPPS. Their interactions with the allosteric pocket of the enzyme were characterized by crystallography, and the results provide further insight into the pharmacophore requirements for allosteric inhibition

Unraveling the Prenylation–Cancer Paradox in Multiple Myeloma with Novel Geranylgeranyl Pyrophosphate Synthase (GGPPS) Inhibitors

Post-translational prenylation of the small GTP-binding proteins (GTPases) is vital to a plethora of biological processes, including cellular proliferation. We have identified a new class of thienopyrimidine-based bisphosphonate (ThP-BP) inhibitors of the human geranylgeranyl pyrophosphate synthase (hGGPPS) that block protein prenylation in multiple myeloma (MM) cells leading to cellular apoptosis. These inhibitors are also effective in blocking the proliferation of other types of cancer cells. We confirmed intracellular target engagement, demonstrated the mechanism of action leading to apoptosis, and determined a direct correlation between apoptosis and intracellular inhibition of hGGPPS. Administration of a ThP-BP inhibitor to a MM mouse model confirmed in vivo downregulation of Rap1A geranylgeranylation and reduction of monoclonal immunoglobulins (M-protein, a biomarker of disease burden) in the serum. These results provide the first proof-of-principle that hGGPPS is a valuable therapeutic target in oncology and more specifically for the treatment of multiple myeloma