22 results on '"Santos, Webster L."'
Search Results
2. Targeting negative energy balance with calorie restriction and mitochondrial uncoupling in db/db mice.
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Chen, Sing-Young, Beretta, Martina, Olzomer, Ellen M., Shah, Divya P., Wong, Derek Y.H., Alexopoulos, Stephanie J., Aleksovska, Isabella, Salamoun, Joseph M., Garcia, Christopher J., Cochran, Blake J., Rye, Kerry-Anne, Smith, Greg C., Byrne, Frances L., Morris, Margaret J., Santos, Webster L., Cantley, James, and Hoehn, Kyle L.
- Abstract
Calorie restriction is a first-line treatment for overweight individuals with metabolic impairments. However, few patients can adhere to long-term calorie restriction. An alternative approach to calorie restriction that also causes negative energy balance is mitochondrial uncoupling, which decreases the amount of energy that can be extracted from food. Herein we compare the metabolic effects of calorie restriction with the mitochondrial uncoupler BAM15 in the db/db mouse model of severe hyperglycemia, obesity, hypertriglyceridemia, and fatty liver. Male db/db mice were treated with ∼50% calorie restriction, BAM15 at two doses of 0.1% and 0.2% (w/w) admixed in diet, or 0.2% BAM15 with time-restricted feeding from 5 weeks of age. Mice were metabolically phenotyped over 4 weeks with assessment of key readouts including body weight, glucose tolerance, and liver steatosis. At termination, liver tissues were analysed by metabolomics and qPCR. Calorie restriction and high-dose 0.2% BAM15 decreased body weight to a similar extent, but mice treated with BAM15 had far better improvement in glucose control. High-dose BAM15 treatment completely normalized fasting glucose and glucose tolerance to levels similar to lean db/+ control mice. Low-dose 0.1% BAM15 did not affect body mass but partially improved glucose tolerance to a similar degree as 50% calorie restriction. Both calorie restriction and high-dose BAM15 significantly improved hyperglucagonemia and liver and serum triglyceride levels. Combining high-dose BAM15 with time-restricted feeding to match the time that calorie restricted mice were fed resulted in the best metabolic phenotype most similar to lean db/+ controls. BAM15-mediated improvements in glucose control were associated with decreased glucagon levels and decreased expression of enzymes involved in hepatic gluconeogenesis. BAM15 and calorie restriction treatments improved most metabolic disease phenotypes in db/db mice. However, mice fed BAM15 had superior effects on glucose control compared to the calorie restricted group that consumed half as much food. Submaximal dosing with BAM15 demonstrated that its beneficial effects on glucose control are independent of weight loss. These data highlight the potential for mitochondrial uncoupler pharmacotherapies in the treatment of metabolic disease. • Mitochondrial uncoupling and calorie restriction (CR) cause negative energy balance. • This study is first to compare uncoupling and CR in a metabolic disease model. • Both treatments markedly improved metabolic disease phenotypes in db/db mice. • Despite similar weight loss, uncoupling had superior effects on glucose control. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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3. The Positively Charged Surface of Herpes Simplex Virus UL42 Mediates DNA Binding.
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Komazin-Meredith, Gloria, Santos, Webster L., Filman, David J., Hogle, James M., Verdine, Gregory L., and Coen, Donald M.
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HERPES simplex virus , *DNA polymerases , *DNA , *GENETIC mutation , *BIOCHEMISTRY - Abstract
Herpes simplex virus DNA polymerase is a heterodimer composed of UL30, a catalytic subunit, and UL42, a processivity subunit. Mutations that decrease DNA binding by UL42 decrease long chain DNA synthesis by the polymerase. The crystal structure of UL42 bound to the C terminus of UL30 revealed an extensive positively charged surface ("back face"). We tested two hypotheses, 1) the C terminus of UL30 affects DNA binding and 2) the positively charged back face mediates DNA binding. Addressing the first hypothesis, we found that the presence of a peptide corresponding to the UL30 C terminus did not result in altered binding of UL42 to DNA. Addressing the second hypothesis, previous work showed that substitution of four conserved arginine residues on the basic face with alanines resulted in decreased DNA affinity. We tested the affinities for DNA and the stimulation of long chain DNA synthesis of mutants in which the four conserved arginine residues were substituted individually or together with lysines and also a mutant in which a conserved glutamine residue was substituted with an arginine to increase positive charge on the back face. We also engineered cysteines onto this surface to permit disulfide cross-linking studies. Last, we assayed the effects of ionic strength on DNA binding by UL42 to estimate the number of ions released upon binding. Our results taken together strongly suggest that the basic back face of UL42 contacts DNA and that positive charge on this surface is important for this interaction. [ABSTRACT FROM AUTHOR]
- Published
- 2008
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4. Synthesis and biological evaluation of phosphonic and thiophosphoric acid derivatives of lysophosphatidic acid
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Santos, Webster L., Heasley, Brian H., Jarosz, Renata, Carter, Karen M., Lynch, Kevin R., and Macdonald, Timothy L.
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THIOPHOSPHORIC acid , *LYSOPHOSPHOLIPIDS , *PHOSPHATES , *GENES - Abstract
Using an N-oleoyl ethanolamide scaffold, a series of phosphate polar head group analogues of LPA comprised of various α-substituted phosphonates and thiophosphates was prepared. In a broken cell GTP[
γ35 S] binding assay, agonist activity was evaluated at the three LPA receptors of the endothelial differentiation gene (Edg) family. This study has resulted in the discovery of a nonhydrolyzable LPA1-selective agonist (11). Additionally, thiophosphate 19 bears an isosteric phosphate mimetic that confers agonism at the LPA1 receptor but not LPA2. [Copyright &y& Elsevier]- Published
- 2004
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5. Mitochondrial uncoupler SHC517 reverses obesity in mice without affecting food intake.
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Chen, Sing-Young, Beretta, Martina, Alexopoulos, Stephanie J., Shah, Divya P., Olzomer, Ellen M., Hargett, Stefan R., Childress, Elizabeth S., Salamoun, Joseph M., Aleksovska, Isabella, Roseblade, Ariane, Cranfield, Charles, Rawling, Tristan, Quinlan, Kate G.R., Morris, Margaret J., Tucker, Simon P., Santos, Webster L., and Hoehn, Kyle L.
- Subjects
BODY temperature ,FOOD consumption ,HIGH-fat diet ,LEAN body mass ,OBESITY ,RESPIRATORY quotient - Abstract
Mitochondrial uncouplers decrease caloric efficiency and have potential therapeutic benefits for the treatment of obesity and related metabolic disorders. Herein we investigate the metabolic and physiologic effects of a recently identified small molecule mitochondrial uncoupler named SHC517 in a mouse model of diet-induced obesity. SHC517 was administered as an admixture in food. The effect of SHC517 on in vivo energy expenditure and respiratory quotient was determined by indirect calorimetry. A dose-finding obesity prevention study was performed by starting SHC517 treatment concomitant with high fat diet for a period of 12 days. An obesity reversal study was performed by feeding mice western diet for 4 weeks prior to SHC517 treatment for 7 weeks. Biochemical assays were used to determine changes in glucose, insulin, triglycerides, and cholesterol. SHC517 concentrations were determined by mass spectrometry. SHC517 increased lipid oxidation without affecting body temperature. SHC517 prevented diet-induced obesity when administered at 0.05% and 0.1% w /w in high fat diet and reversed established obesity when tested at the 0.05% dose. In the obesity reversal model, SHC517 restored adiposity to levels similar to chow-fed control mice without affecting food intake or lean body mass. SHC517 improved glucose tolerance and fasting glucose levels when administered in both the obesity prevention and obesity reversal modes. SHC517 is a mitochondrial uncoupler with potent anti-obesity and insulin sensitizing effects in mice. SHC517 reversed obesity without altering food intake or compromising lean mass, effects that are highly sought-after in anti-obesity therapeutics. • The mitochondrial uncoupler SHC517 prevents and reverses diet-induced obesity and glucose intolerance in mice. • SHC517 did not alter body temperature, food intake or fat-free lean mass. • SHC517 is a leading candidate for translational development in the field of mitochondrial uncoupler therapeutics for obesity and related diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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6. Discovery and antiparasitic activity of AZ960 as a Trypanosoma brucei ERK8 inhibitor.
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Valenciano, Ana L., Ramsey, Aaron C., Santos, Webster L., and Mackey, Zachary B.
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TRYPANOSOMIASIS treatment , *TRYPANOSOMA brucei , *ANTIPARASITIC agents , *DRUG toxicity , *DRUG prices , *BIOACTIVE compounds , *EXTRACELLULAR signal-regulated kinases , *ENZYME inhibitors - Abstract
Human African trypanosomiasis (HAT) is a lethal, vector-borne disease caused by the parasite Trypanosoma brucei . Therapeutic strategies for this neglected tropical disease suffer from disadvantages such as toxicity, high cost, and emerging resistance. Therefore, new drugs with novel modes of action are needed. We screened cultured T. brucei against a focused kinase inhibitor library to identify promising bioactive compounds. Among the ten hits identified from the phenotypic screen, AZ960 emerged as the most promising compound with potent antiparasitic activity (IC 50 = 120 nM) and was shown to be a selective inhibitor of an essential gene product, T. brucei extracellular signal-regulated kinase 8 (TbERK8). We report that AZ960 has a K i of 1.25 μM for TbERK8 and demonstrate its utility in establishing TbERK8 as a potentially druggable target in T. brucei . [ABSTRACT FROM AUTHOR]
- Published
- 2016
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7. Mapping Targetable Sites on Human Telomerase RNA Pseudoknot/Template Domain Using 2'-OMe RNA-interacting Polynucleotide (RIPtide) Microarrays.
- Author
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Gude, Lourdes, Berkovitch, Shaunna S., Santos, Webster L., Kutchukian, Peter S., Pawloski, Adam R., Kuimelis, Robert, McGall, Glenn, and Verdine, Gregory L.
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TELOMERASE , *NUCLEIC acids , *PROTEIN microarrays , *LIGANDS (Biochemistry) , *OLIGONUCLEOTIDES - Abstract
Most cellular RNAs engage in intrastrand base-pairing that gives rise to complex three-dimensional folds. This self-pairing presents an impediment toward binding of the RNA by nucleic acid-based ligands. An important step in the discovery of RNAtargeting ligands is therefore to identify those regions in a folded RNA that are accessible toward the nucleic acid-based ligand. Because the folding of RNA targets can involve interactions between nonadjacent regions and employ both Watson-Crick and non-Watson-Crick base-pairing, screening of candidate binder ensembles is typically necessary. Microarray-based screening approaches have shown great promise in this regard and have suggested that achieving complete sequence coverage would be a valuable attribute of a next generation system. Here, we report a custom microarray displaying a library of RNA-interacting polynucleotides comprising all possible 2'-OMe RNA sequences from 4- to 8-nucleotides in length. We demonstrate the utility of this array in identifying RNA-interacting polynucleotides that bind tightly and specifically to the highly conserved, functionally essential template/pseudoknot domain of human telomerase RNA and that inhibit telomerase function in vitro. [ABSTRACT FROM AUTHOR]
- Published
- 2012
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8. The Human Cytomegalovirus UL44 C Clamp Wraps around DNA
- Author
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Komazin-Meredith, Gloria, Petrella, Robert J., Santos, Webster L., Filman, David J., Hogle, James M., Verdine, Gregory L., Karplus, Martin, and Coen, Donald M.
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DNA polymerases , *POLYMERASE chain reaction , *CYTOMEGALOVIRUSES , *CROSSLINKING (Polymerization) - Abstract
Summary: Processivity factors tether the catalytic subunits of DNA polymerases to DNA so that continuous synthesis of long DNA strands is possible. The human cytomegalovirus DNA polymerase subunit UL44 forms a C clamp-shaped dimer intermediate in structure between monomeric herpes simplex virus UL42, which binds DNA directly via a basic surface, and the trimeric sliding clamp PCNA, which encircles DNA. To investigate how UL44 interacts with DNA, calculations were performed in which a 12 bp DNA oligonucleotide was docked to UL44. The calculations suggested that UL44 encircles DNA, which interacts with basic residues both within the cavity of the C clamp and in flexible loops of UL44 that complete the “circle.” The results of mutational and crosslinking studies were consistent with this model. Thus, UL44 is a “hybrid” of UL42 and PCNA: its structure is intermediate between the two and its mode of interaction with DNA has elements of both. [Copyright &y& Elsevier]
- Published
- 2008
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9. Imidazole-based sphingosine-1-phosphate transporter Spns2 inhibitors.
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Shrader, Christopher W., Foster, Daniel, Kharel, Yugesh, Huang, Tao, Lynch, Kevin R., and Santos, Webster L.
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SPHINGOSINE-1-phosphate , *STRUCTURE-activity relationships , *CONCENTRATION gradient , *LYMPHOID tissue , *HELA cells , *IMIDAZOLES , *AMPA receptors - Abstract
[Display omitted] Sphingosine-1-phosphate (S1P) is a chemotactic lipid that influences immune cell positioning. S1P concentration gradients are necessary for proper egress of lymphocytes from the thymus and secondary lymphoid tissues. This trafficking is interdicted by S1P receptor modulators, and it is expected that S1P transporter (Spns2) inhibitors, by reshaping S1P concentration gradients, will do the same. We previously reported SLF1081851 as a prototype Spns2 inhibitor, which provided a scaffold to investigate the importance of the oxadiazole core and the terminal amine. In this report, we disclose a structure–activity relationship study by incorporating imidazole as both a linker and surrogate for a positive charge in SLF1081851. In vitro inhibition of Spns2-dependent S1P transport in HeLa cells identified 7b as an inhibitor with an IC 50 of 1.4 ± 0.3 µM. The SAR studies reported herein indicate that imidazolium can be a substitute for the terminal amine in SLF1081851 and that Spns2 inhibition is highly dependent on the lipid alkyl tail length. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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10. Molecular recognition of a branched peptide with HIV-1 Rev Response Element (RRE) RNA.
- Author
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Dai, Yumin, Peralta, Ashley N., Wynn, Jessica E., Sherpa, Chringma, Li, Hao, Verma, Astha, Le Grice, Stuart F.J., and Santos, Webster L.
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RNA , *MOLECULAR recognition , *BINDING sites , *LEAD compounds - Abstract
Graphical abstract Abstract Interaction of HIV-1 rev response element (RRE) RNA with its cognate protein, Rev, is critical for HIV-1 replication. Understanding the mode of interaction between RRE RNA and ligands at the binding site can facilitate RNA molecular recognition as well as provide a strategy for developing anti-HIV therapeutics. Our approach utilizes branched peptides as a scaffold for multivalent binding to RRE IIB (high affinity rev binding site) with incorporation of unnatural amino acids to increase affinity via non-canonical interactions with the RNA. Previous high throughput screening of a 46,656-member library revealed several hits that bound RRE IIB RNA in the sub-micromolar range. In particular, the lead compound, 4B3, displayed a K d value of 410 nM and demonstrated selectivity towards RRE. A ribonuclease protection assay revealed that 4B3 binds to the stem-loop structure of RRE IIB RNA, which was confirmed by SHAPE analysis with 234 nt long NL4-3 RRE RNA. Our studies further indicated interaction of 4B3 with both primary and secondary Rev binding sites. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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11. Semireduction of alkynoic acids via a transition metal-free α borylation-protodeborylation sequence.
- Author
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Verma, Astha, Grams, R. Justin, Rastatter, Brett P., and Santos, Webster L.
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ACRYLIC acid , *ACIDS - Abstract
Abstract A method for the semi-reduction of alkynoic acids through an α-borylation and subsequent protodeborylation mechanism has been developed. The transition metal-free protocol is achieved through the activation of bis(pinacolato)diboron by an in situ generated carboxylate moiety yielding aryl acrylic acids. Our studies demonstrate an unprecedented dual role for the carboxylate anion that involves the activation of the diboron reagent and a directing effect in the α-borylation. Graphical abstract Image 1 [ABSTRACT FROM AUTHOR]
- Published
- 2019
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12. Characterization and in vitro activity of a branched peptide boronic acid that interacts with HIV-1 RRE RNA.
- Author
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Wynn, Jessica E., Zhang, Wenyu, Tebit, Denis M., Gray, Laurie R., Hammarskjold, Marie-Louise, Rekosh, David, and Santos, Webster L.
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BORONIC acids , *BINDING sites , *RNA analysis , *IN vitro studies , *HIV - Abstract
A branched peptide containing multiple boronic acids was found to bind RRE IIB selectively and inhibit HIV-1 p24 capsid production in a dose-dependent manner. Structure–activity relationship studies revealed that branching in the peptide is crucial for the low micromolar binding towards RRE IIB, and the peptide demonstrates selectivity towards RRE IIB in the presence of tRNA. Footprinting studies suggest a binding site on the upper stem and internal loop regions of the RNA, which induces enzymatic cleavage of the internal loops of RRE IIB upon binding. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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13. Conversion of oxadiazolo[3,4-b]pyrazines to imidazo[4,5-b]pyrazines via a tandem reduction-cyclization sequence generates new mitochondrial uncouplers.
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Dai, Yumin, Santiago-Rivera, José A., Hargett, Stefan, Salamoun, Joseph M., Hoehn, Kyle L., and Santos, Webster L.
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MITOCHONDRIA , *PYRAZINES , *IMIDAZOLES , *RING formation (Chemistry) , *CYTOCHROME oxidase - Abstract
[Display omitted] We report new mitochondrial uncouplers derived from the conversion of [1,2,5]oxadiazolo[3,4- b ]pyrazines to 1 H -imidazo[4,5- b ]pyrazines. The in situ Fe-mediated reduction of the oxadiazole fragment followed by cyclization gave access to imidazopyrazines in moderate to good yields. A selection of orthoesters also allowed functionalization on the 2-position of the imidazole ring. This method afforded a variety of imidazopyrazine derivatives with varying substitution on the 2, 5 and 6 positions. Our studies suggest that both a 2-trifluoromethyl group and N -methylation are crucial for mitochondrial uncoupling capacity. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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14. Structure–activity relationship studies of the lipophilic tail region of sphingosine kinase 2 inhibitors.
- Author
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Congdon, Molly D., Childress, Elizabeth S., Patwardhan, Neeraj N., Gumkowski, James, Morris, Emily A., Kharel, Yugesh, Lynch, Kevin R., and Santos, Webster L.
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STRUCTURE-activity relationship in pharmacology , *SPHINGOSINE kinase , *ENZYME inhibitors , *DRUG lipophilicity , *PHENYL group - Abstract
Sphingosine-1-phosphate (S1P) is a ubiquitous, endogenous small molecule that is synthesized by two isoforms of sphingosine kinase (SphK1 and 2). Intervention of the S1P signaling pathway has attracted significant attention because alteration of S1P levels is linked to several disease states including cancer, fibrosis, and sickle cell disease. While intense investigations have focused on developing SphK1 inhibitors, only a limited number of SphK2-selective agents have been reported. Herein, we report our investigations on the structure–activity relationship studies of the lipophilic tail region of SLR080811 , a SphK2-selective inhibitor. Our studies demonstrate that the internal phenyl ring is a key structural feature that is essential in the SLR080811 scaffold. Further, we show the dependence of SphK2 activity and selectivity on alkyl tail length, suggesting a larger lipid binding pocket in SphK2 compared to SphK1. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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15. Structure–activity relationships of furazano[3,4-b]pyrazines as mitochondrial uncouplers.
- Author
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Kenwood, Brandon M., Calderone, Joseph A., Taddeo, Evan P., Hoehn, Kyle L., and Santos, Webster L.
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STRUCTURE-activity relationship in pharmacology , *PYRAZINES , *MITOCHONDRIAL pathology , *ADENOSINE triphosphatase , *COMPARATIVE studies - Abstract
Chemical mitochondrial uncouplers are lipophilic weak acids that transport protons into the mitochondrial matrix via a pathway that is independent of ATP synthase, thereby uncoupling nutrient oxidation from ATP production. These uncouplers have potential for the treatment of diseases such as obesity, Parkinson’s disease, and aging. We have previously identified a novel mitochondrial protonophore, named BAM15, which stimulates mitochondrial respiration across a broad dosing range compared to carbonyl cyanide p -trifluoromethoxyphenylhydrazone (FCCP). Herein, we report our investigations on the structure–activity relationship profile of BAM15. Our studies demonstrate the importance of the furazan, pyrazine, and aniline rings as well as p K a in maintaining its effective protonophore activity. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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16. Effect of alkyl chain length on sphingosine kinase 2 selectivity
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Knott, Kenneth, Kharel, Yugesh, Raje, Mithun R., Lynch, Kevin R., and Santos, Webster L.
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SPHINGOSINE kinase , *ALKYL group , *SPHINGOSINE-1-phosphate , *CANCER treatment , *FIBROSIS , *ENZYME inhibitors , *STRUCTURE-activity relationship in pharmacology - Abstract
Abstract: The conversion of sphingosine to sphingosine-1-phosphate is catalyzed by sphingosine kinase (SphK), which has been implicated in disease states such as cancer and fibrosis. Because SphK exists as two different isoforms, SphK1 and SphK2, understanding the physiological function of each isoenzyme is important. Of the two isoenzymes, SphK2 is significantly less understood, which is evident by the lack of selective small molecule inhibitors. Building on our initial work that focused on the structure–activity relationship study on an FTY720-derived cylohexylamine scaffold, we report that varying the alkyl chain length on the hydrophobic tail can impart selectivity toward SphK2 over SphK1. [Copyright &y& Elsevier]
- Published
- 2012
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17. Design, synthesis and biological activity of sphingosine kinase 2 selective inhibitors
- Author
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Raje, Mithun R., Knott, Kenneth, Kharel, Yugesh, Bissel, Philippe, Lynch, Kevin R., and Santos, Webster L.
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SPHINGOSINE-1-phosphate , *CANCER cells , *PHOSPHORYLATION , *ENZYME inhibitors , *TARGETED drug delivery , *ANTINEOPLASTIC agents , *TUMOR growth - Abstract
Abstract: Sphingosine kinase (SphK) has emerged as an attractive target for cancer therapeutics due to its role in cell survival. SphK phosphorylates sphingosine to form sphingosine 1-phosphate (S1P), which has been implicated in cancer growth and survival. SphK exists as two different isotypes, namely SphK1 and SphK2, which play different roles inside the cell. In this report, we describe SphK inhibitors based on the immunomodulatory drug, FTY720, which is phosphorylated by SphK2 to generate a S1P mimic. Structural modification of FTY720 provided a template for synthesizing new inhibitors. A diversity-oriented synthesis generated a library of SphK inhibitors with a novel scaffold and headgroup. We have discovered subtype selective inhibitors with K i’s in the low micromolar range. This is the first report describing quaternary ammonium salts as SphK inhibitors. [Copyright &y& Elsevier]
- Published
- 2012
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18. Subunit-specific Protein Footprinting Reveals Significant Structural Rearrangements and a Role for N-terminal Lys-14 of HIV-1 Integrase during Viral DNA Binding.
- Author
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Zhuojun Zhao, McKee, Christopher J., Kessl, Jacques J., Santos, Webster L., Daigle, Janet E., Engelman, Alan, Verdine, Gregory, and Kvaratskhelia, Mamuka
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MICROBIAL enzymes , *HIV , *MASS spectrometry , *DNA , *NUCLEOPROTEINS , *CONFORMATIONAL analysis - Abstract
To identify functional contacts between HIV-1 integrase (IN) and its viral DNA substrate, we devised a new experimental strategy combining the following two methodologies. First, disulfide-mediated cross-linking was used to site-specifically link select core and C-terminal domain amino acids to respective positions in viral DNA. Next, surface topologies of free IN and IN-DNA complexes were compared using Lys- and Arg-selective small chemical modifiers and mass spectrometric analysis. This approach enabled us to dissect specific contacts made by different monomers within the multimeric complex. The foot-printing studies for the first time revealed the importance of a specific N-terminal domain residue, Lys-14, in viral DNA binding. In addition, a DNA-induced conformational change involving the connection between the core and C-terminal domains was observed. Site-directed mutagenesis experiments confirmed the importance of the identified contacts for recombinant IN activities and virus infection. These new findings provided major constraints, enabling us to identify the viral DNA binding channel in the active full-length IN multimer. The experimental approach described here has general application to mapping inter actions within functional nucleoprotein complexes. [ABSTRACT FROM AUTHOR]
- Published
- 2008
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19. Reactivity of atropaldehyde, a felbamate metabolite in human liver tissue in vitro
- Author
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Kapetanovic, Izet M., Torchin, Cynthia D., Strong, John M., Yonekawa, Wayne D., Lu, Chuang, Li, Albert P., Dieckhaus, Christine M., Santos, Webster L., Macdonald, Timothy L., Sofia, R. Duane, and Kupferberg, Harvey J.
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BIOTRANSFORMATION (Metabolism) , *ACROLEIN ,SIDE effects of anticonvulsants - Abstract
Antiepileptic therapy with a broad spectrum drug felbamate (FBM) has been limited due to reports of hepatotoxicity and aplastic anemia associated with its use. It was proposed that a bioactivation of FBM leading to formation of α,β-unsaturated aldehyde, atropaldehyde (ATPAL) could be responsible for toxicities associated with the parent drug. Other members of this class of compounds, acrolein and 4-hydroxynonenal (HNE), are known for their reactivity and toxicity. It has been proposed that the bioactivation of FBM to ATPAL proceeds though a more stable cyclized product, 4-hydroxy-5-phenyltetrahydro-1,3-oxazin-2-one (CCMF) whose formation has been shown recently. Aldehyde dehydrogenase (ALDH) and glutathione transferase (GST) are detoxifying enzymes and targets for reactive aldehydes. This study examined effects of ATPAL and its precursor, CCMF on ALDH, GST and cell viability in liver, the target tissue for its metabolism and toxicity. A known toxin, HNE, which is also a substrate for ALDH and GST, was used for comparison. Interspecies difference in metabolism of FBM is well documented, therefore, human tissue was deemed most relevant and used for these studies. ATPAL inhibited ALDH and GST activities and led to a loss of hepatocyte viability. Several fold greater concentrations of CCMF were necessary to demonstrate a similar degree of ALDH inhibition or cytotoxicity as observed with ATPAL. This is consistent with CCMF requiring prior conversion to the more proximate toxin, ATPAL. GSH was shown to protect against ALDH inhibition by ATPAL. In this context, ALDH and GST are detoxifying pathways and their inhibition would lead to an accumulation of reactive species from FBM metabolism and/or metabolism of other endogenous or exogenous compounds and predisposing to or causing toxicity. Therefore, mechanisms of reactive aldehydes toxicity could include direct interaction with critical cellular macromolecules or indirect interference with cellular detoxification mechanisms. [Copyright &y& Elsevier]
- Published
- 2002
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20. Probing the substitution pattern of indole-based scaffold reveals potent and selective sphingosine kinase 2 inhibitors.
- Author
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Congdon, Molly, Fritzemeier, Russell G., Kharel, Yugesh, Brown, Anne M., Serbulea, Vlad, Bevan, David R., Lynch, Kevin R., and Santos, Webster L.
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SPHINGOSINE kinase , *KINASE inhibitors , *INDOLE , *HYDROGEN bonding , *MOLECULAR models , *BINDING sites - Abstract
Elevated levels of sphingosine 1-phosphate (S1P) and increased expression of sphingosine kinase isoforms (SphK1 and SphK2) have been implicated in a variety of disease states including cancer, inflammation, and autoimmunity. Consequently, the S1P signaling axis has become an attractive target for drug discovery. Selective inhibition of either SphK1 or SphK2 has been demonstrated to be effective in modulating S1P levels in animal models. While SphK1 inhibitors have received much attention, the development of potent and selective SphK2 inhibitors are emerging. Previously, our group reported a SphK2 naphthalene-based selective inhibitor, SLC5081308 , which displays approximately 7-fold selectivity for hSphK2 over hSphK1 and has a SphK2 K i value of 1.0 μM. To improve SphK2 potency and selectivity, we designed, synthesized, and evaluated a series of indole-based compounds derived from SLC5081308. After investigating substitution patterns around the indole ring, we discovered that 1,5-disubstitution promoted optimal binding in the SphK2 substrate binding site and subsequent inhibition of enzymatic activity. Our studies led to the identification of SLC5101465 (6r , SphK2 K i = 90 nM, >110 fold selective for SphK2 over SphK1). Molecular modeling studies revealed key nonpolar interactions with Val308, Phe548, His556, and Cys533 and hydrogen bonds with both Asp211 and Asp308 as responsible for the high SphK2 inhibition and selectivity. Image 1 • New indole-based sphingosine kinase 2 inhibitors were designed and synthesized. • SLC5101465 (6r) showed potent inhibition of SphK2 (K i = 90 nM) and selectivity over SphK1 (>110 fold). • SLC5101465 (6r) was validated to inhibit SphK2 in vitro in U937 cells as well as in S. cerevisiae yeast assay. • Molecular modeling revealed key nonpolar interactions within the binding pocket and hydrogen bonds with Asp211 and Asp308. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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21. Lipophilic tail modifications of 2-(hydroxymethyl)pyrrolidine scaffold reveal dual sphingosine kinase 1 and 2 inhibitors.
- Author
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Li, Hao, Sibley, Christopher D., Kharel, Yugesh, Huang, Tao, Brown, Anne M., Wonilowicz, Laura G., Bevan, David R., Lynch, Kevin R., and Santos, Webster L.
- Subjects
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SPHINGOSINE kinase , *HYDROXYMETHYL compounds , *PYRROLIDINE , *DIFFUSE large B-cell lymphomas , *STRUCTURE-activity relationships , *DITHIOCARBAMATES - Abstract
The sphingosine 1-phosphate (S1P) signaling pathway is an attractive target for pharmacological manipulation due to its involvement in cancer progression and immune cell chemotaxis. The synthesis of S1P is catalyzed by the action of sphingosine kinase 1 or 2 (SphK1 or SphK2) on sphingosine and ATP. While potent and selective inhibitors of SphK1 or SphK2 have been reported, development of potent dual SphK1/SphK2 inhibitors are still needed. Towards this end, we report the structure–activity relationship profiling of 2-(hydroxymethyl)pyrrolidine-based inhibitors with 22d being the most potent dual SphK1/SphK2 inhibitor (SphK1 K i = 0.679 μM, SphK2 K i = 0.951 μM) reported in this series. 22d inhibited the growth of engineered Saccharomyces cerevisiae and decreased S1P levels in histiocytic lymphoma myeloid cell line (U937 cells), demonstrating inhibition of SphK1 and 2 in vitro. Molecular modeling studies of 22d docked inside the Sph binding pocket of both SphK1 and SphK2 indicate essential hydrogen bond between the 2-(hydroxymethyl)pyrrolidine head to interact with aspartic acid and serine residues near the ATP binding pocket, which provide the basis for dual inhibition. In addition, the dodecyl tail adopts a "J-shape" conformation found in crystal structure of sphingosine bound to SphK1. Collectively, these studies provide insight into the intermolecular interactions in the SphK1 and 2 active sites to achieve maximal dual inhibitory activity. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
22. Anilinopyrazines as potential mitochondrial uncouplers.
- Author
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Murray, Jacob H., Hargett, Stefan, Hoehn, Kyle L., and Santos, Webster L.
- Subjects
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FATTY liver , *MITOCHONDRIAL membranes , *STRUCTURE-activity relationships , *NEURODEGENERATION - Abstract
Mitochondrial protonophores transport protons through the mitochondrial inner membrane into the matrix to uncouple nutrient oxidation from ATP production thereby decreasing the proton motive force. Mitochondrial uncouplers have beneficial effects of decrease reactive oxygen species generation and have the potential for treating diseases such as obesity, neurodegenerative diseases, non-alcoholic fatty liver disease (NAFLD), diabetes, and many others. In this study, we report the structure-activity relationship profile of the pyrazine scaffold bearing substituted aniline rings. Our work indicates that a trifluoromethyl group is best at the para position while the trifluoromethoxy group is preferred in the meta position of the aniline rings of 2,3-substituted pyrazines. As proton transport and cycling requires the formation of a negative charge that has to traverse the mitochondrial membrane, a stabilizing internal hydrogen bond is a key feature for efficient mitochondrial uncoupling activity. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
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