Limits...
Asymmetric synthesis and evaluation of danshensu-cysteine conjugates as novel potential anti-apoptotic drug candidates.

Pan LL, Wang J, Jia YL, Zheng HM, Wang Y, Zhu YZ - Int J Mol Sci (2014)

Bottom Line: We have previously reported that the danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) is a potent anti-oxidative and anti-apoptotic agent.Herein, we further design and asymmetrically synthesize two diastereoisomers of DSC and explore their potential bioactivities.Our results provide strong evidence that DSC and its two diastereoisomers have similar anti-oxidative activity and that DSC exerts significant vascular-protective effects, at least in part, through inhibition of apoptosis and modulation of endogenous antioxidant enzymes.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Bioactive Small Molecules, Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai 201203, China. panlilong@fudan.edu.cn.

ABSTRACT
We have previously reported that the danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) is a potent anti-oxidative and anti-apoptotic agent. Herein, we further design and asymmetrically synthesize two diastereoisomers of DSC and explore their potential bioactivities. Our results show that DSC and its two diastereoisomers exert similar protective effects in hydrogen peroxide (H2O2)-induced cellular injury in SH-SY5Y cells, as evidenced by the increase of cell viability, superoxide dismutase (SOD), and reduced glutathione (GSH) activity, and glutathione peroxidase (GPx) expression, and the decrease of cellular morphological changes and nuclear condensation, lactate dehydrogenase (LDH) release, and malondialdehyde (MDA) production. In H2O2-stimulated human umbilical vein endothelial cells (HUVEC), DSC concentration-dependently attenuates H2O2-induced cell death, LDH release, mitochondrial membrane potential collapse, and modulates the expression of apoptosis-related proteins (Bcl-2, Bax, caspase-3, and caspase-9). Our results provide strong evidence that DSC and its two diastereoisomers have similar anti-oxidative activity and that DSC exerts significant vascular-protective effects, at least in part, through inhibition of apoptosis and modulation of endogenous antioxidant enzymes.

Show MeSH

Related in: MedlinePlus

DSC modulated apoptosis-related proteins expression in H2O2-stimulated HUVEC. HUVEC were incubated with indicated concentrations of DSC or NAC for 4 h, then stimulated with H2O2 (200 μM) for 12 h; the Bcl-2 (A); Bax (B); caspase-3 (C); and caspase-9 (D) were measured by Western blot, respectively. Western blot and densitometric analysis for Bcl-2 (A); Bax (B); caspase-3 (C); and caspase-9 (D). β-actin was used as loading control. Protein expression in control cells was set to 1. Data shown are means ± SEM, &p < 0.05 compared with unstimulated cells, *p < 0.05 compared with H2O2-stimulated cells; Data were from at least three independent experiments, each performed in duplicate.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4307265&req=5

ijms-16-00628-f008: DSC modulated apoptosis-related proteins expression in H2O2-stimulated HUVEC. HUVEC were incubated with indicated concentrations of DSC or NAC for 4 h, then stimulated with H2O2 (200 μM) for 12 h; the Bcl-2 (A); Bax (B); caspase-3 (C); and caspase-9 (D) were measured by Western blot, respectively. Western blot and densitometric analysis for Bcl-2 (A); Bax (B); caspase-3 (C); and caspase-9 (D). β-actin was used as loading control. Protein expression in control cells was set to 1. Data shown are means ± SEM, &p < 0.05 compared with unstimulated cells, *p < 0.05 compared with H2O2-stimulated cells; Data were from at least three independent experiments, each performed in duplicate.

Mentions: To further demonstrate the anti-apoptotic activity of DSC on H2O2-stimulated HUVEC, apoptosis-related proteins were analyzed by Western blot. As shown in Figure 8A,B, H2O2 stimulation significantly decreased Bcl-2 expression and increased Bax expression level. However, pretreatment with DSC concentration-dependently increased Bcl-2 expression and decreased Bax expression in H2O2-stimulated HUVEC. Meanwhile, H2O2 stimulation markedly induced degradation of caspase-3 and caspase-9 (Figure 8C,D). Consistent with our previous study [6], DSC concentration-dependently inhibited H2O2-induced caspase-3 and caspase-9 degradation in HUVEC. In addition, NAC treatment also attenuated H2O2-mediated changes of apoptosis-related proteins expression.


Asymmetric synthesis and evaluation of danshensu-cysteine conjugates as novel potential anti-apoptotic drug candidates.

Pan LL, Wang J, Jia YL, Zheng HM, Wang Y, Zhu YZ - Int J Mol Sci (2014)

DSC modulated apoptosis-related proteins expression in H2O2-stimulated HUVEC. HUVEC were incubated with indicated concentrations of DSC or NAC for 4 h, then stimulated with H2O2 (200 μM) for 12 h; the Bcl-2 (A); Bax (B); caspase-3 (C); and caspase-9 (D) were measured by Western blot, respectively. Western blot and densitometric analysis for Bcl-2 (A); Bax (B); caspase-3 (C); and caspase-9 (D). β-actin was used as loading control. Protein expression in control cells was set to 1. Data shown are means ± SEM, &p < 0.05 compared with unstimulated cells, *p < 0.05 compared with H2O2-stimulated cells; Data were from at least three independent experiments, each performed in duplicate.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307265&req=5

ijms-16-00628-f008: DSC modulated apoptosis-related proteins expression in H2O2-stimulated HUVEC. HUVEC were incubated with indicated concentrations of DSC or NAC for 4 h, then stimulated with H2O2 (200 μM) for 12 h; the Bcl-2 (A); Bax (B); caspase-3 (C); and caspase-9 (D) were measured by Western blot, respectively. Western blot and densitometric analysis for Bcl-2 (A); Bax (B); caspase-3 (C); and caspase-9 (D). β-actin was used as loading control. Protein expression in control cells was set to 1. Data shown are means ± SEM, &p < 0.05 compared with unstimulated cells, *p < 0.05 compared with H2O2-stimulated cells; Data were from at least three independent experiments, each performed in duplicate.
Mentions: To further demonstrate the anti-apoptotic activity of DSC on H2O2-stimulated HUVEC, apoptosis-related proteins were analyzed by Western blot. As shown in Figure 8A,B, H2O2 stimulation significantly decreased Bcl-2 expression and increased Bax expression level. However, pretreatment with DSC concentration-dependently increased Bcl-2 expression and decreased Bax expression in H2O2-stimulated HUVEC. Meanwhile, H2O2 stimulation markedly induced degradation of caspase-3 and caspase-9 (Figure 8C,D). Consistent with our previous study [6], DSC concentration-dependently inhibited H2O2-induced caspase-3 and caspase-9 degradation in HUVEC. In addition, NAC treatment also attenuated H2O2-mediated changes of apoptosis-related proteins expression.

Bottom Line: We have previously reported that the danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) is a potent anti-oxidative and anti-apoptotic agent.Herein, we further design and asymmetrically synthesize two diastereoisomers of DSC and explore their potential bioactivities.Our results provide strong evidence that DSC and its two diastereoisomers have similar anti-oxidative activity and that DSC exerts significant vascular-protective effects, at least in part, through inhibition of apoptosis and modulation of endogenous antioxidant enzymes.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Bioactive Small Molecules, Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai 201203, China. panlilong@fudan.edu.cn.

ABSTRACT
We have previously reported that the danshensu-cysteine conjugate N-((R)-3-benzylthio-1-methoxy-1-oxo-2-propanyl)-2-acetoxy-3-(3,4-diacetoxyphenyl) propanamide (DSC) is a potent anti-oxidative and anti-apoptotic agent. Herein, we further design and asymmetrically synthesize two diastereoisomers of DSC and explore their potential bioactivities. Our results show that DSC and its two diastereoisomers exert similar protective effects in hydrogen peroxide (H2O2)-induced cellular injury in SH-SY5Y cells, as evidenced by the increase of cell viability, superoxide dismutase (SOD), and reduced glutathione (GSH) activity, and glutathione peroxidase (GPx) expression, and the decrease of cellular morphological changes and nuclear condensation, lactate dehydrogenase (LDH) release, and malondialdehyde (MDA) production. In H2O2-stimulated human umbilical vein endothelial cells (HUVEC), DSC concentration-dependently attenuates H2O2-induced cell death, LDH release, mitochondrial membrane potential collapse, and modulates the expression of apoptosis-related proteins (Bcl-2, Bax, caspase-3, and caspase-9). Our results provide strong evidence that DSC and its two diastereoisomers have similar anti-oxidative activity and that DSC exerts significant vascular-protective effects, at least in part, through inhibition of apoptosis and modulation of endogenous antioxidant enzymes.

Show MeSH
Related in: MedlinePlus