Cysteine proteases continue to provide validated targets for treatment of human diseases.In neurodegenerative disorders,multiple cysteine proteases provide targets for enzyme inhibitors,notably caspases,calpains,and c...Cysteine proteases continue to provide validated targets for treatment of human diseases.In neurodegenerative disorders,multiple cysteine proteases provide targets for enzyme inhibitors,notably caspases,calpains,and cathepsins.The reactive,active-site cysteine provides specificity for many inhibitor designs over other families of proteases,such as aspartate and serine;however,a)inhibitor strategies often use covalent enzyme modification,and b)obtaining selectivity within families of cysteine proteases and their isozymes is problematic.This review provides a general update on strategies for cysteine protease inhibitor design and a focus on cathepsin B and calpain 1 as drug targets for neurodegenerative disorders;the latter focus providing an interesting query for the contemporary assumptions that irreversible,covalent protein modification and low selectivity are anathema to therapeutic safety and efficacy.展开更多
BACKGROUND:Increasing evidence suggests that the inactivation of cathepsin B attenuates hepatocyte apoptosis and liver damage.This study aimed to investigate the protective effects of a cathepsin B inhibitor(CA-074me)...BACKGROUND:Increasing evidence suggests that the inactivation of cathepsin B attenuates hepatocyte apoptosis and liver damage.This study aimed to investigate the protective effects of a cathepsin B inhibitor(CA-074me) on lipopolysaccharide(LPS)/D-galactosamine(D-GalN)-induced acute hepatic failure(AHF) in mice.METHODS:Mice were intraperitoneally injected with a combination of LPS/D-GalN to induce AHF with or without CA-074me pretreatment.The cumulative survival rates were calculated 48 hours after the induction of AHF.As well as changes in biochemical indicators and liver histology,hepatocyte apoptosis was assessed using a TUNEL method.Serum tumor necrosis factor-α(TNF-α) production,caspase-3,caspase-8,and caspase-9 activity was evaluated.Cytosolic cytochrome c and Bcl-2 expression were measured by Western blotting.RESULTS:The marked elevation in serum aminotransferase activity and prothrombin time found in LPS/D-GalN-treated mice was significantly improved by pretreatment with CA074me.The efficacy of CA-074me was also confirmed by histological analysis and TUNEL assay.The survival rate significantly improved in LPS/D-GalN-induced mice given CA-074me compared with untreated mice.LPS/D-GalNinduced caspase-3 and caspase-9 activation was remarkably suppressed by CA-074me.However,the increased levels of serum TNF-α and elevated caspase-8 activity in AHF mice were not significantly reduced by CA-074me.Moreover,CA074me sharply reduced the increased expression of cytosolic cytochrome c and markedly augmented Bcl-2 expression.CONCLUSION:These results suggest that CA-074me has a protective effect in acute hepatic failure induced by LPS/D-GalN.展开更多
Objective To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs. Methods To obtain the purified cathepsin K, we cloned the target fragment fro...Objective To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs. Methods To obtain the purified cathepsin K, we cloned the target fragment from the mRNA of human osteosacoma cell line MG63 and demonstrated its correctness through DNA sequencing. Cathepsin K was expressed in a high amount in E.coli after IPTG induction, and was purified to near homogenetity through resolution and column purification. The specificity of the protein was shown by Western blotting experiment. The biological activity of the components in the fermentation broth was assayed by their inhibitory effects on cathepsin K and its analog papain. Results With the inhibition of papain activity as a screen index, the fermentation samples of one thousand strains of fungi were tested and 9 strains among them showed strong inhibitory effects. The crude products of the fermentation broth were tested for their specific inhibitory effects on the purified human cathepsin K, the product of fungi 2358 shows the highest specificity against cathepsin K. Conclusions The compounds isolated from fungi 2358 show the highest biological activity and are worth further structure elucidation and function characterization.展开更多
目的:探讨参一胶囊联合一线化疗方案与单独化疗治疗不能手术的晚期IIIb/Ⅳ初治非小细胞肺癌(NSCLC)患者血清基质金属蛋白酶9(MMP-9)及组织蛋白酶抑制剂-1(TIMP-1)水平变化,并观察其近期疗效和治疗后不良反应。方法:采用酶联免疫吸附法(E...目的:探讨参一胶囊联合一线化疗方案与单独化疗治疗不能手术的晚期IIIb/Ⅳ初治非小细胞肺癌(NSCLC)患者血清基质金属蛋白酶9(MMP-9)及组织蛋白酶抑制剂-1(TIMP-1)水平变化,并观察其近期疗效和治疗后不良反应。方法:采用酶联免疫吸附法(ELISA)检测89例肺癌患者化疗前后血浆MMP-9及TIMP-1水平。结果:两组有效率无显著性差异(P>0.05),两组疾控率比较差异有显著性(P<0.05)。全部CR+PR、SD、PD患者治疗前血清MMP-9、TIMP-1水平比较,无显著性差异(P>0.05),治疗后差异均具有有显著性(P<0.05),随疗效降低呈升高趋势。两组相比疗前与疗后两次血清MMP-9水平,参一联合化疗组下降更明显,差异有显著性(P<0.05),而血清TIMP-1水平差异无显著性(P>0.05)。IV期患者比IIIb期患者外周血清MMP-9水平明显增高,差异有显著性(P<0.05)。但外周血清TIMP-1在IIIb期和IV期无统计学差异。其他因素如年龄、性别、ECOG评分、病理与MMP-9、TIMP-1水平均未发现显著性差异。患者治疗后出现常见不良反应及生活质量(quality of life,QOL)观察:观察组(即参一胶囊联合化疗组)在胃肠道反应(如恶心、呕吐、腹胀等),骨髓抑制(如白细胞减少、贫血、血小板下降等)方面均较单独化疗组减轻(P<0.05),参一联合组在改善QOL方面也明显高于对照组,且有统计学差异(P<0.05)。结论:参一胶囊联合化疗组患者外周血清MMP-9水平下降更明显,且有统计学差异,而患者血清TIMP-1水平变化无统计学差异。血清MMP-9、TIMP-1水平治疗后随疗效降低呈升高趋势,因此,两者对患者疗效判定具有一定的参考价值。两组外周血清MMP-9、TIMP-1水平变化不受性别、年龄、ECOG评分影响。肺癌患者TNM分期对NSCLC患者外周血清MMP-9水平变化有影响,分期越晚,其外周血清MMP-9水平值越高。因此,MMP-9血清值越高,提示预后越差。外周血清TIMP-1水平与MMP-9水平在化疗前后NSCLC患者呈正相关。参一胶囊对非小细胞肺癌患者可以增加疗效,减少毒副反应,生活质量得到提高,不良反应如消化道症状及骨髓抑制均可以减轻。展开更多
文摘Cysteine proteases continue to provide validated targets for treatment of human diseases.In neurodegenerative disorders,multiple cysteine proteases provide targets for enzyme inhibitors,notably caspases,calpains,and cathepsins.The reactive,active-site cysteine provides specificity for many inhibitor designs over other families of proteases,such as aspartate and serine;however,a)inhibitor strategies often use covalent enzyme modification,and b)obtaining selectivity within families of cysteine proteases and their isozymes is problematic.This review provides a general update on strategies for cysteine protease inhibitor design and a focus on cathepsin B and calpain 1 as drug targets for neurodegenerative disorders;the latter focus providing an interesting query for the contemporary assumptions that irreversible,covalent protein modification and low selectivity are anathema to therapeutic safety and efficacy.
文摘BACKGROUND:Increasing evidence suggests that the inactivation of cathepsin B attenuates hepatocyte apoptosis and liver damage.This study aimed to investigate the protective effects of a cathepsin B inhibitor(CA-074me) on lipopolysaccharide(LPS)/D-galactosamine(D-GalN)-induced acute hepatic failure(AHF) in mice.METHODS:Mice were intraperitoneally injected with a combination of LPS/D-GalN to induce AHF with or without CA-074me pretreatment.The cumulative survival rates were calculated 48 hours after the induction of AHF.As well as changes in biochemical indicators and liver histology,hepatocyte apoptosis was assessed using a TUNEL method.Serum tumor necrosis factor-α(TNF-α) production,caspase-3,caspase-8,and caspase-9 activity was evaluated.Cytosolic cytochrome c and Bcl-2 expression were measured by Western blotting.RESULTS:The marked elevation in serum aminotransferase activity and prothrombin time found in LPS/D-GalN-treated mice was significantly improved by pretreatment with CA074me.The efficacy of CA-074me was also confirmed by histological analysis and TUNEL assay.The survival rate significantly improved in LPS/D-GalN-induced mice given CA-074me compared with untreated mice.LPS/D-GalNinduced caspase-3 and caspase-9 activation was remarkably suppressed by CA-074me.However,the increased levels of serum TNF-α and elevated caspase-8 activity in AHF mice were not significantly reduced by CA-074me.Moreover,CA074me sharply reduced the increased expression of cytosolic cytochrome c and markedly augmented Bcl-2 expression.CONCLUSION:These results suggest that CA-074me has a protective effect in acute hepatic failure induced by LPS/D-GalN.
文摘Objective To establish an effective assay to access the effects of natural products on cathepsin K for screening antiosteoporosis drugs. Methods To obtain the purified cathepsin K, we cloned the target fragment from the mRNA of human osteosacoma cell line MG63 and demonstrated its correctness through DNA sequencing. Cathepsin K was expressed in a high amount in E.coli after IPTG induction, and was purified to near homogenetity through resolution and column purification. The specificity of the protein was shown by Western blotting experiment. The biological activity of the components in the fermentation broth was assayed by their inhibitory effects on cathepsin K and its analog papain. Results With the inhibition of papain activity as a screen index, the fermentation samples of one thousand strains of fungi were tested and 9 strains among them showed strong inhibitory effects. The crude products of the fermentation broth were tested for their specific inhibitory effects on the purified human cathepsin K, the product of fungi 2358 shows the highest specificity against cathepsin K. Conclusions The compounds isolated from fungi 2358 show the highest biological activity and are worth further structure elucidation and function characterization.
文摘目的:探讨参一胶囊联合一线化疗方案与单独化疗治疗不能手术的晚期IIIb/Ⅳ初治非小细胞肺癌(NSCLC)患者血清基质金属蛋白酶9(MMP-9)及组织蛋白酶抑制剂-1(TIMP-1)水平变化,并观察其近期疗效和治疗后不良反应。方法:采用酶联免疫吸附法(ELISA)检测89例肺癌患者化疗前后血浆MMP-9及TIMP-1水平。结果:两组有效率无显著性差异(P>0.05),两组疾控率比较差异有显著性(P<0.05)。全部CR+PR、SD、PD患者治疗前血清MMP-9、TIMP-1水平比较,无显著性差异(P>0.05),治疗后差异均具有有显著性(P<0.05),随疗效降低呈升高趋势。两组相比疗前与疗后两次血清MMP-9水平,参一联合化疗组下降更明显,差异有显著性(P<0.05),而血清TIMP-1水平差异无显著性(P>0.05)。IV期患者比IIIb期患者外周血清MMP-9水平明显增高,差异有显著性(P<0.05)。但外周血清TIMP-1在IIIb期和IV期无统计学差异。其他因素如年龄、性别、ECOG评分、病理与MMP-9、TIMP-1水平均未发现显著性差异。患者治疗后出现常见不良反应及生活质量(quality of life,QOL)观察:观察组(即参一胶囊联合化疗组)在胃肠道反应(如恶心、呕吐、腹胀等),骨髓抑制(如白细胞减少、贫血、血小板下降等)方面均较单独化疗组减轻(P<0.05),参一联合组在改善QOL方面也明显高于对照组,且有统计学差异(P<0.05)。结论:参一胶囊联合化疗组患者外周血清MMP-9水平下降更明显,且有统计学差异,而患者血清TIMP-1水平变化无统计学差异。血清MMP-9、TIMP-1水平治疗后随疗效降低呈升高趋势,因此,两者对患者疗效判定具有一定的参考价值。两组外周血清MMP-9、TIMP-1水平变化不受性别、年龄、ECOG评分影响。肺癌患者TNM分期对NSCLC患者外周血清MMP-9水平变化有影响,分期越晚,其外周血清MMP-9水平值越高。因此,MMP-9血清值越高,提示预后越差。外周血清TIMP-1水平与MMP-9水平在化疗前后NSCLC患者呈正相关。参一胶囊对非小细胞肺癌患者可以增加疗效,减少毒副反应,生活质量得到提高,不良反应如消化道症状及骨髓抑制均可以减轻。
