This work describes our efforts to optimize the lead PI3Kα inhibitor N-benzyl 4-hydroxy-2-quinolo... more This work describes our efforts to optimize the lead PI3Kα inhibitor N-benzyl 4-hydroxy-2-quinolone-3-carboxamide using structure-based design and molecular docking. We identified a series of N-phenyl 4-hydroxy-2-quinolone-3-carboxamides as selective inhibitors of mutant H1047R versus wild-type PI3Kα and we also showed that the cell growth inhibition by these compounds likely occurs by inhibiting the formation of pAKT and induction of apoptosis.
Objective. The treatment of phenylketonuria (PKU) in children and adults has been difficult becau... more Objective. The treatment of phenylketonuria (PKU) in children and adults has been difficult because of erosion of dietary adherence, leading to poor school performance, impairment of executive functioning, loss of IQ, and deterioration of white matter in the brain. Mutant PKU mice produced by exposure to N-ethyl-N′-nitrosourea (ENU) were used to examine the effect of large neutral amino acid (LNAA) supplementation on brain and blood phenylalanine (Phe). Methods. Mice with PKU, genotype ENU 2/2 with features of classical PKU, were supplemented with LNAA while on a normal diet. Two dosages of LNAA were given 0.5 g/kg and 1.0 g/kg by gavage. Blood Phe was determined in the experimental, control, and sham-treated mice. Brain Phe was determined by magnetic resonance spectroscopy after perchloric acid extraction. Branched-chain amino acid transferase (BCAT) was determined in brain as a marker for energy metabolism. Results. Blood Phe was reduced in the LNAA-treated mice by an average of 1...
The utilization of the long range optimized proton-carbon chemical shift correlation experiment i... more The utilization of the long range optimized proton-carbon chemical shift correlation experiment in the total assignment of the proton and carbon nmr spectra of 9-methylphenanthro[4,3-a]dibenzothiophene is described. The experiment was employed in concert with conventional proton-carbon chemical shift correlation, heteronuclear relayed coherence transfer and broad band homonuclearly decoupled proton-carbon chemical shift correlation experiments. The experiments in combination offer a useful alternative to the 13C-13C double quantum coherence experiments, providing significantly better sensitivity and nearly the same capabilities.
To evaluate the modulatory effects of trace metals on lymphocyte growth and maturation, thymidine... more To evaluate the modulatory effects of trace metals on lymphocyte growth and maturation, thymidine uptake (TU), protein, ATP, Fe, Cu, Zn, ferritin, CD3, CD4, CD8 antigens, surface transferrin receptors (TFR) and interleukin 2 receptors (IL2R) were assessed in normal and T cell leukemia human lymphocytes, cultured in media with varying Fe, Cu and Zn concentrations [Me]. In normal lymphocytes in media with optimal [Me], all values increased significantly after PHA stimulation, except for intracellular metal concentration and CD3+, CD4+, CD8+ cells which were unchanged. In media with low or high [Me], all parameters except for CD8+ cells were decreased. In unstimulated ALL lymphocytes grown in media with optimal [Me], TU, protein, ATP, CD4+, Fe, Cu and ferritin were higher and Zn and CD8+ lower than in unstimulated normal cells: they did not change after PHA stimulation, except in media with low [Me], in which they approached the values of stimulated normal lymphocytes. TFR and IL2R for...
... Leland L. Smith* and Edward L. Ezell Department of Human Biological Chemistry and Genetics, U... more ... Leland L. Smith* and Edward L. Ezell Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, Texas 77550, USA ... Holden-Day, San Francisco (1964); (b) L. Toscano, G. Guadagnini and G. Grisanti, Arzneim. forsch. ...
500 MHz and especially 750 MHz 1 H NMR spectra of the hydrogenated fullerenes C 60 H 2 and C 60 H... more 500 MHz and especially 750 MHz 1 H NMR spectra of the hydrogenated fullerenes C 60 H 2 and C 60 H 4 reveal much unexpected fine structure that apparently results from the partial alignment of the molecules in the magnetic field. Molecules with an anisotropic magnetic ...
... EDWARD L. EZEU and LELAND L. SMITH* Deparfiwit of Hrmari Biological Cheiuisrr) and Grnrfirs. ... more ... EDWARD L. EZEU and LELAND L. SMITH* Deparfiwit of Hrmari Biological Cheiuisrr) and Grnrfirs. ... More recently, C spectra of 1 and 10-hydroxycamptothecin [5] have been reported by Lin and Cordell (9), with the suggestion that the earlier 'k-nmr data for 1 needed revision. ...
Journal of Labelled Compounds and Radiopharmaceuticals, 1996
Page 1. Journal of Labelled Compounds and Radiopharmaceuticals-Vol. XYXVIII, No. 8 Stabilities of... more Page 1. Journal of Labelled Compounds and Radiopharmaceuticals-Vol. XYXVIII, No. 8 Stabilities of 3H-and 2H-labelled Camptothecins, Hellmuth R. Hinz' Nicholas J. Harris' Beppino C. Giovanella', Edward L. Ezell' and Joachim G. LiehrJe4 ...
ABSTRACT Homonuclear two-dimensional J-resolved (2DJ) spectra of molecules containing strongly co... more ABSTRACT Homonuclear two-dimensional J-resolved (2DJ) spectra of molecules containing strongly coupled spin systems contain additional responses and thus additional information, the responses arising from the mixing effects of the 180° pulse employed to create the spin echo upon which the experiment is based. Although additional responses due to strong coupling are generally considered to be an undesirable complication inherent to the experiment, it is shown that they can serve a useful correlation function. Total assignment of the 1H-nmr spectrum of 2-(2′-pyridyl)-1,8-naphthyridine at 100 MHz is reported, and members of the three strongly coupled spin systems of the molecule are identified via the additional responses due to strong coupling.
LL-23 is a natural peptide corresponding to the 23 N-terminal amino acid residues of human host d... more LL-23 is a natural peptide corresponding to the 23 N-terminal amino acid residues of human host defense cathelicidin LL-37. LL-23 demonstrated, compared to LL-37, a conserved ability to induce the chemokine MCP-1 in human peripheral blood mononuclear cells, a lack of ability to suppress induction of the pro-inflammatory cytokine TNF-α in response to bacterial lipopolysaccharides (LPS), and reduced antimicrobial activity. Heteronuclear multidimensional nuclear magnetic resonance (NMR) characterization of LL-23 revealed similar secondary structures and backbone dynamics in three membrane-mimetic micelles: SDS, dodecylphosphocholine (DPC), and dioctanoylphosphatidylglycerol. The NMR structure of LL-23 determined in perdeuterated DPC contained a unique serine that segregated the hydrophobic surface of the amphipathic helix into two domains. To improve our understanding, Ser9 of LL-23was changed to either Ala or Val on the basis of homologous primate cathelicidins. These changes made the hydrophobic surface of LL-23 continuous and enhanced antibacterial activity. While identical helical structures did not explain the altered activities, a reduced rate of hydrogen-deuterium exchange from LL-23 to LL-23A9 to LL-23V9 suggested a deeper penetration of LL-23V9 into the interior of the micelles, which correlated with enhanced activities. Moreover, these LL-23 variants had discrete immunomodulatory activities. Both restored the TNF-α dampening activity to the level of LL-37. Furthermore, LL-23A9, like LL-23, maintained superior protective MCP-1 production, while LL-23V9 was strongly immunosuppressive, preventing baseline MCP-1 induction and substantially reducing LPS-stimulated MCP-1 production. Thus, these LL-23 variants, designed on the basis of a structural hot spot, are promising immune modulators that are easier to synthesize and less toxic to mammalian cells than the parent peptide LL-37.
The treatment of phenylketonuria (PKU) in children and adults has been difficult because of erosi... more The treatment of phenylketonuria (PKU) in children and adults has been difficult because of erosion of dietary adherence, leading to poor school performance, impairment of executive functioning, loss of IQ, and deterioration of white matter in the brain. Mutant PKU mice produced by exposure to N-ethyl-N'-nitrosourea (ENU) were used to examine the effect of large neutral amino acid (LNAA) supplementation on brain and blood phenylalanine (Phe). Mice with PKU, genotype ENU 2/2 with features of classical PKU, were supplemented with LNAA while on a normal diet. Two dosages of LNAA were given 0.5 g/kg and 1.0 g/kg by gavage. Blood Phe was determined in the experimental, control, and sham-treated mice. Brain Phe was determined by magnetic resonance spectroscopy after perchloric acid extraction. Branched-chain amino acid transferase (BCAT) was determined in brain as a marker for energy metabolism. Blood Phe was reduced in the LNAA-treated mice by an average of 15% (0.5 g/kg) and 50% (1....
Quercetin is highly mutagenic in vitro, yet is not carcinogenic when administered chronically at ... more Quercetin is highly mutagenic in vitro, yet is not carcinogenic when administered chronically at large doses to rodents for 12 months. We hypothesized that catechol-O-methyltransferase-catalyzed O-methylation of quercetin and other mutagenic catechol-containing flavonoids may provide an efficient inactivation in vivo and may therefore prevent tumor induction by these flavonoids. After one intraperitoneal administration of 50 mg/kg quercetin to hamsters, a urinary ether extract contained 2% quercetin and 97% 3'-O-methylquercetin. When the urine was treated first with beta-glucuronidase and sulfatase, 13% quercetin and 87% 3'-O-methylquercetin were recovered. Quercetin was rapidly O-methylated by either porcine liver or hamster kidney catechol-O-methyltransferase, with Km values of 6.1 and 6.9 microM and Vmax values of 14,870 and 200 pmol/mg of protein/min, respectively. S-Adenosyl-L-homocysteine exhibited a potent feedback inhibition of the catechol-O-methyltransferase-cataly...
When praziquantel was exposed to N-bromosuccinimide in the presence of ethanol, a tricyclic 3-bro... more When praziquantel was exposed to N-bromosuccinimide in the presence of ethanol, a tricyclic 3-bromo-1-ethoxy pyrazinone was formed. From this and the analogous 1,3-dibromopyrazinone, a small library of 3-alkylamino-1-ethoxy, 1,3-dialkoxy, 3-alkoxy-1-bromo, and 3-alkylamino-1-bromo substituted pyrazinones were synthesized in high yields.
Creatine ethyl ester hydrochloride (CEE) was synthesized as a prodrug of creatine (CRT) to improv... more Creatine ethyl ester hydrochloride (CEE) was synthesized as a prodrug of creatine (CRT) to improve aqueous solubility, gastrointestinal permeability, and ultimately the pharmacodynamics of CRT. We used high-performance liquid chromatography (HPLC) and proton nuclear magnetic resonance (NMR) to characterize the pH-dependent stability of CEE in aqueous solution and compared the permeability of CEE to CRT and creatinine (CRN) across Caco-2 human epithelial cell monolayers and transdermal permeability across porcine skin. CEE was most stable in a strongly acidic condition (half-life = 570 hours at pH 1.0) where it undergoes ester hydrolysis to CRT and ethanol. At pH ≥ 1.0, CEE cyclizes to CRN with the logarithm of the first order rate constant increasing linearly with pH. Above pH 8.0 (half-life = 23 sec) the rate of degradation was too rapid to be determined. The rate of degradation of CEE in cell culture media and simulated intestinal fluid (SIF) was a function of pH and correlated we...
This work describes our efforts to optimize the lead PI3Kα inhibitor N-benzyl 4-hydroxy-2-quinolo... more This work describes our efforts to optimize the lead PI3Kα inhibitor N-benzyl 4-hydroxy-2-quinolone-3-carboxamide using structure-based design and molecular docking. We identified a series of N-phenyl 4-hydroxy-2-quinolone-3-carboxamides as selective inhibitors of mutant H1047R versus wild-type PI3Kα and we also showed that the cell growth inhibition by these compounds likely occurs by inhibiting the formation of pAKT and induction of apoptosis.
Objective. The treatment of phenylketonuria (PKU) in children and adults has been difficult becau... more Objective. The treatment of phenylketonuria (PKU) in children and adults has been difficult because of erosion of dietary adherence, leading to poor school performance, impairment of executive functioning, loss of IQ, and deterioration of white matter in the brain. Mutant PKU mice produced by exposure to N-ethyl-N′-nitrosourea (ENU) were used to examine the effect of large neutral amino acid (LNAA) supplementation on brain and blood phenylalanine (Phe). Methods. Mice with PKU, genotype ENU 2/2 with features of classical PKU, were supplemented with LNAA while on a normal diet. Two dosages of LNAA were given 0.5 g/kg and 1.0 g/kg by gavage. Blood Phe was determined in the experimental, control, and sham-treated mice. Brain Phe was determined by magnetic resonance spectroscopy after perchloric acid extraction. Branched-chain amino acid transferase (BCAT) was determined in brain as a marker for energy metabolism. Results. Blood Phe was reduced in the LNAA-treated mice by an average of 1...
The utilization of the long range optimized proton-carbon chemical shift correlation experiment i... more The utilization of the long range optimized proton-carbon chemical shift correlation experiment in the total assignment of the proton and carbon nmr spectra of 9-methylphenanthro[4,3-a]dibenzothiophene is described. The experiment was employed in concert with conventional proton-carbon chemical shift correlation, heteronuclear relayed coherence transfer and broad band homonuclearly decoupled proton-carbon chemical shift correlation experiments. The experiments in combination offer a useful alternative to the 13C-13C double quantum coherence experiments, providing significantly better sensitivity and nearly the same capabilities.
To evaluate the modulatory effects of trace metals on lymphocyte growth and maturation, thymidine... more To evaluate the modulatory effects of trace metals on lymphocyte growth and maturation, thymidine uptake (TU), protein, ATP, Fe, Cu, Zn, ferritin, CD3, CD4, CD8 antigens, surface transferrin receptors (TFR) and interleukin 2 receptors (IL2R) were assessed in normal and T cell leukemia human lymphocytes, cultured in media with varying Fe, Cu and Zn concentrations [Me]. In normal lymphocytes in media with optimal [Me], all values increased significantly after PHA stimulation, except for intracellular metal concentration and CD3+, CD4+, CD8+ cells which were unchanged. In media with low or high [Me], all parameters except for CD8+ cells were decreased. In unstimulated ALL lymphocytes grown in media with optimal [Me], TU, protein, ATP, CD4+, Fe, Cu and ferritin were higher and Zn and CD8+ lower than in unstimulated normal cells: they did not change after PHA stimulation, except in media with low [Me], in which they approached the values of stimulated normal lymphocytes. TFR and IL2R for...
... Leland L. Smith* and Edward L. Ezell Department of Human Biological Chemistry and Genetics, U... more ... Leland L. Smith* and Edward L. Ezell Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, Texas 77550, USA ... Holden-Day, San Francisco (1964); (b) L. Toscano, G. Guadagnini and G. Grisanti, Arzneim. forsch. ...
500 MHz and especially 750 MHz 1 H NMR spectra of the hydrogenated fullerenes C 60 H 2 and C 60 H... more 500 MHz and especially 750 MHz 1 H NMR spectra of the hydrogenated fullerenes C 60 H 2 and C 60 H 4 reveal much unexpected fine structure that apparently results from the partial alignment of the molecules in the magnetic field. Molecules with an anisotropic magnetic ...
... EDWARD L. EZEU and LELAND L. SMITH* Deparfiwit of Hrmari Biological Cheiuisrr) and Grnrfirs. ... more ... EDWARD L. EZEU and LELAND L. SMITH* Deparfiwit of Hrmari Biological Cheiuisrr) and Grnrfirs. ... More recently, C spectra of 1 and 10-hydroxycamptothecin [5] have been reported by Lin and Cordell (9), with the suggestion that the earlier 'k-nmr data for 1 needed revision. ...
Journal of Labelled Compounds and Radiopharmaceuticals, 1996
Page 1. Journal of Labelled Compounds and Radiopharmaceuticals-Vol. XYXVIII, No. 8 Stabilities of... more Page 1. Journal of Labelled Compounds and Radiopharmaceuticals-Vol. XYXVIII, No. 8 Stabilities of 3H-and 2H-labelled Camptothecins, Hellmuth R. Hinz' Nicholas J. Harris' Beppino C. Giovanella', Edward L. Ezell' and Joachim G. LiehrJe4 ...
ABSTRACT Homonuclear two-dimensional J-resolved (2DJ) spectra of molecules containing strongly co... more ABSTRACT Homonuclear two-dimensional J-resolved (2DJ) spectra of molecules containing strongly coupled spin systems contain additional responses and thus additional information, the responses arising from the mixing effects of the 180° pulse employed to create the spin echo upon which the experiment is based. Although additional responses due to strong coupling are generally considered to be an undesirable complication inherent to the experiment, it is shown that they can serve a useful correlation function. Total assignment of the 1H-nmr spectrum of 2-(2′-pyridyl)-1,8-naphthyridine at 100 MHz is reported, and members of the three strongly coupled spin systems of the molecule are identified via the additional responses due to strong coupling.
LL-23 is a natural peptide corresponding to the 23 N-terminal amino acid residues of human host d... more LL-23 is a natural peptide corresponding to the 23 N-terminal amino acid residues of human host defense cathelicidin LL-37. LL-23 demonstrated, compared to LL-37, a conserved ability to induce the chemokine MCP-1 in human peripheral blood mononuclear cells, a lack of ability to suppress induction of the pro-inflammatory cytokine TNF-α in response to bacterial lipopolysaccharides (LPS), and reduced antimicrobial activity. Heteronuclear multidimensional nuclear magnetic resonance (NMR) characterization of LL-23 revealed similar secondary structures and backbone dynamics in three membrane-mimetic micelles: SDS, dodecylphosphocholine (DPC), and dioctanoylphosphatidylglycerol. The NMR structure of LL-23 determined in perdeuterated DPC contained a unique serine that segregated the hydrophobic surface of the amphipathic helix into two domains. To improve our understanding, Ser9 of LL-23was changed to either Ala or Val on the basis of homologous primate cathelicidins. These changes made the hydrophobic surface of LL-23 continuous and enhanced antibacterial activity. While identical helical structures did not explain the altered activities, a reduced rate of hydrogen-deuterium exchange from LL-23 to LL-23A9 to LL-23V9 suggested a deeper penetration of LL-23V9 into the interior of the micelles, which correlated with enhanced activities. Moreover, these LL-23 variants had discrete immunomodulatory activities. Both restored the TNF-α dampening activity to the level of LL-37. Furthermore, LL-23A9, like LL-23, maintained superior protective MCP-1 production, while LL-23V9 was strongly immunosuppressive, preventing baseline MCP-1 induction and substantially reducing LPS-stimulated MCP-1 production. Thus, these LL-23 variants, designed on the basis of a structural hot spot, are promising immune modulators that are easier to synthesize and less toxic to mammalian cells than the parent peptide LL-37.
The treatment of phenylketonuria (PKU) in children and adults has been difficult because of erosi... more The treatment of phenylketonuria (PKU) in children and adults has been difficult because of erosion of dietary adherence, leading to poor school performance, impairment of executive functioning, loss of IQ, and deterioration of white matter in the brain. Mutant PKU mice produced by exposure to N-ethyl-N'-nitrosourea (ENU) were used to examine the effect of large neutral amino acid (LNAA) supplementation on brain and blood phenylalanine (Phe). Mice with PKU, genotype ENU 2/2 with features of classical PKU, were supplemented with LNAA while on a normal diet. Two dosages of LNAA were given 0.5 g/kg and 1.0 g/kg by gavage. Blood Phe was determined in the experimental, control, and sham-treated mice. Brain Phe was determined by magnetic resonance spectroscopy after perchloric acid extraction. Branched-chain amino acid transferase (BCAT) was determined in brain as a marker for energy metabolism. Blood Phe was reduced in the LNAA-treated mice by an average of 15% (0.5 g/kg) and 50% (1....
Quercetin is highly mutagenic in vitro, yet is not carcinogenic when administered chronically at ... more Quercetin is highly mutagenic in vitro, yet is not carcinogenic when administered chronically at large doses to rodents for 12 months. We hypothesized that catechol-O-methyltransferase-catalyzed O-methylation of quercetin and other mutagenic catechol-containing flavonoids may provide an efficient inactivation in vivo and may therefore prevent tumor induction by these flavonoids. After one intraperitoneal administration of 50 mg/kg quercetin to hamsters, a urinary ether extract contained 2% quercetin and 97% 3'-O-methylquercetin. When the urine was treated first with beta-glucuronidase and sulfatase, 13% quercetin and 87% 3'-O-methylquercetin were recovered. Quercetin was rapidly O-methylated by either porcine liver or hamster kidney catechol-O-methyltransferase, with Km values of 6.1 and 6.9 microM and Vmax values of 14,870 and 200 pmol/mg of protein/min, respectively. S-Adenosyl-L-homocysteine exhibited a potent feedback inhibition of the catechol-O-methyltransferase-cataly...
When praziquantel was exposed to N-bromosuccinimide in the presence of ethanol, a tricyclic 3-bro... more When praziquantel was exposed to N-bromosuccinimide in the presence of ethanol, a tricyclic 3-bromo-1-ethoxy pyrazinone was formed. From this and the analogous 1,3-dibromopyrazinone, a small library of 3-alkylamino-1-ethoxy, 1,3-dialkoxy, 3-alkoxy-1-bromo, and 3-alkylamino-1-bromo substituted pyrazinones were synthesized in high yields.
Creatine ethyl ester hydrochloride (CEE) was synthesized as a prodrug of creatine (CRT) to improv... more Creatine ethyl ester hydrochloride (CEE) was synthesized as a prodrug of creatine (CRT) to improve aqueous solubility, gastrointestinal permeability, and ultimately the pharmacodynamics of CRT. We used high-performance liquid chromatography (HPLC) and proton nuclear magnetic resonance (NMR) to characterize the pH-dependent stability of CEE in aqueous solution and compared the permeability of CEE to CRT and creatinine (CRN) across Caco-2 human epithelial cell monolayers and transdermal permeability across porcine skin. CEE was most stable in a strongly acidic condition (half-life = 570 hours at pH 1.0) where it undergoes ester hydrolysis to CRT and ethanol. At pH ≥ 1.0, CEE cyclizes to CRN with the logarithm of the first order rate constant increasing linearly with pH. Above pH 8.0 (half-life = 23 sec) the rate of degradation was too rapid to be determined. The rate of degradation of CEE in cell culture media and simulated intestinal fluid (SIF) was a function of pH and correlated we...
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