Background Amyotrophic lateral sclerosis(ALS)is a progressive neurodegenerative disease that begins with motor neuron death in the spinal cord and cerebral cortex,ultimately resulting in death from respiratory distres...Background Amyotrophic lateral sclerosis(ALS)is a progressive neurodegenerative disease that begins with motor neuron death in the spinal cord and cerebral cortex,ultimately resulting in death from respiratory distress(breathing failure).About 90%of ALS cases are sporadic,and 10%of ALS cases are of the inherited type with a genetic cause.About 150 different gene mutations have been reported so far.SOD1 is a well-identified gene associated with ALS.Indeed,SOD1 aggregation has been reported in ALS patients,but the mechanism of SOD1 aggregation remains unclear.Our previous work showed that inhibiting SOD1 aggregation with a hit compound(PRG-A-01)could reduce the SOD1-induced cytotoxicity and extend the lifespan of ALS mouse model(SOD1G93A−Tg).However,the low bioavailability and rapid degradation of the compound in vivo necessitates the development of a more effective candidate.We generated different derivatives and finally obtained the most potential drug candidate,PRG-A-04.Methods Neuronal cell lines were transfected with the mutant SOD1 expression vector and incubated with PRGA-04.SOD1 aggregation was examined by SOD1 oligomerization assay,immunofluorescence and dot blot assay.The interaction between GST-conjugated SOD1 recombinant proteins and PRG-A-04 was identified using LC–MS/MS and GST pull-down assay.To check the in vivo therapeutic effect of PRG-A-04,SOD1G93A−Tg mice were injected with PRG-A-04;then behavioral test,histological analysis and microarray were performed.Results PRG-A-04 demonstrated favorable pharmacokinetics including high bioavailability and significant blood–brain barrier penetration.Indeed,oral administration of PRG-A-04 in ALS mouse model inhibited the aggregation of SOD1 in the spinal cord,protected against neuronal loss,and extended the lifespan of ALS mice by up to 3 weeks.In vitro,PRG-A-04 selectively bound to the mutant form of SOD1,but not the wild type,and efficiently inhibited the aggregation caused by SOD1-G147P(a SOD1 trimer stabilizer).Conclusions Our findings underscore the potential of targeting trimeric SOD1 in ALS treatment,positioning PRG-A-04 as a strong drug candidate for both familial and sporadic ALS.展开更多
基金supported by the Bio&Medical Technology Development Program of the National Research Foundation of Korea(NRF)grants funded by the Korean government(MSIT)(RS-2024-00399681)the National Research Foundation of Korea(NRF)grants funded by the Korean government(MSIT)(RS-2024-00339289)the Korea Drug Development Fund funded by Ministry of Science and ICT,Ministry of Trade,Industry,and Energy,and Ministry of Health and Welfare(RS-2023-00258714).
文摘Background Amyotrophic lateral sclerosis(ALS)is a progressive neurodegenerative disease that begins with motor neuron death in the spinal cord and cerebral cortex,ultimately resulting in death from respiratory distress(breathing failure).About 90%of ALS cases are sporadic,and 10%of ALS cases are of the inherited type with a genetic cause.About 150 different gene mutations have been reported so far.SOD1 is a well-identified gene associated with ALS.Indeed,SOD1 aggregation has been reported in ALS patients,but the mechanism of SOD1 aggregation remains unclear.Our previous work showed that inhibiting SOD1 aggregation with a hit compound(PRG-A-01)could reduce the SOD1-induced cytotoxicity and extend the lifespan of ALS mouse model(SOD1G93A−Tg).However,the low bioavailability and rapid degradation of the compound in vivo necessitates the development of a more effective candidate.We generated different derivatives and finally obtained the most potential drug candidate,PRG-A-04.Methods Neuronal cell lines were transfected with the mutant SOD1 expression vector and incubated with PRGA-04.SOD1 aggregation was examined by SOD1 oligomerization assay,immunofluorescence and dot blot assay.The interaction between GST-conjugated SOD1 recombinant proteins and PRG-A-04 was identified using LC–MS/MS and GST pull-down assay.To check the in vivo therapeutic effect of PRG-A-04,SOD1G93A−Tg mice were injected with PRG-A-04;then behavioral test,histological analysis and microarray were performed.Results PRG-A-04 demonstrated favorable pharmacokinetics including high bioavailability and significant blood–brain barrier penetration.Indeed,oral administration of PRG-A-04 in ALS mouse model inhibited the aggregation of SOD1 in the spinal cord,protected against neuronal loss,and extended the lifespan of ALS mice by up to 3 weeks.In vitro,PRG-A-04 selectively bound to the mutant form of SOD1,but not the wild type,and efficiently inhibited the aggregation caused by SOD1-G147P(a SOD1 trimer stabilizer).Conclusions Our findings underscore the potential of targeting trimeric SOD1 in ALS treatment,positioning PRG-A-04 as a strong drug candidate for both familial and sporadic ALS.
