Development of KEAP1-targeting PROTAC and its antioxidant properties: In vitro and in vivo
Oxidative stress due to abnormal accumulation of reactive oxygen species (ROS) is an initiator of a large number of human diseases, and thus, the elimination and prevention of excessive ROS are important aspects of preventing the development of such diseases. Nuclear factor erythroid 2-related facto...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-08-01
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| Series: | Redox Biology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231723001842 |
| _version_ | 1797784925447913472 |
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| author | Se Yong Park Raju Gurung Jung Ho Hwang Ju-Hee Kang Hyun Jin Jung Alam Zeb Jong-Ik Hwang Sung Jean Park Han-Joo Maeng Dongyun Shin Seung Hyun Oh |
| author_facet | Se Yong Park Raju Gurung Jung Ho Hwang Ju-Hee Kang Hyun Jin Jung Alam Zeb Jong-Ik Hwang Sung Jean Park Han-Joo Maeng Dongyun Shin Seung Hyun Oh |
| author_sort | Se Yong Park |
| collection | DOAJ |
| description | Oxidative stress due to abnormal accumulation of reactive oxygen species (ROS) is an initiator of a large number of human diseases, and thus, the elimination and prevention of excessive ROS are important aspects of preventing the development of such diseases. Nuclear factor erythroid 2-related factor 2 (NRF2) is an essential transcription factor that defends against oxidative stress, and its function is negatively controlled by Kelch-like ECH-associated protein 1 (KEAP1). Therefore, activating NRF2 by inhibiting KEAP1 is viewed as a strategy for combating oxidative stress-related diseases. Here, we generated a cereblon (CRBN)-based proteolysis-targeting chimera (PROTAC), which we named SD2267, that induces the proteasomal degradation of KEAP1 and leads to NRF2 activation. As was intended, SD2267 bound to KEAP1, recruited CRBN, and induced the degradation of KEAP1. Furthermore, the KEAP1 degradation efficacy of SD2267 was diminished by MG132 (a proteasomal degradation inhibitor) but not by chloroquine (an autophagy inhibitor), which suggested that KEAP1 degradation by SD2267 was proteasomal degradation-dependent and autophagy-independent. Following KEAP1 degradation, SD2267 induced the nuclear translocation of NRF2, which led to the expression of NRF2 target genes and attenuated ROS accumulation induced by acetaminophen (APAP) in hepatocytes. Based on in vivo pharmacokinetic study, SD2267 was injected intraperitoneally at 1 or 3 mg/kg in APAP-induced liver injury mouse model. We observed that SD2267 degraded hepatic KEAP1 and attenuated APAP-induced liver damage. Summarizing, we described the synthesis of a KEAP1-targeting PROTAC (SD2267) and its efficacy and mode of action in vitro and in vivo. The results obtained suggest that SD2267 could be used to treat hepatic diseases related to oxidative stress. |
| first_indexed | 2024-03-13T00:46:48Z |
| format | Article |
| id | doaj.art-c914f61dd15f42f592f9ec9242d4d5fa |
| institution | Directory Open Access Journal |
| issn | 2213-2317 |
| language | English |
| last_indexed | 2024-03-13T00:46:48Z |
| publishDate | 2023-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj.art-c914f61dd15f42f592f9ec9242d4d5fa2023-07-09T04:21:52ZengElsevierRedox Biology2213-23172023-08-0164102783Development of KEAP1-targeting PROTAC and its antioxidant properties: In vitro and in vivoSe Yong Park0Raju Gurung1Jung Ho Hwang2Ju-Hee Kang3Hyun Jin Jung4Alam Zeb5Jong-Ik Hwang6Sung Jean Park7Han-Joo Maeng8Dongyun Shin9Seung Hyun Oh10College of Veterinary Medicine, Seoul National University, Seoul, Republic of KoreaCollege of Pharmacy, Gachon University, Incheon, Republic of KoreaCollege of Pharmacy, Gachon University, Incheon, Republic of KoreaCollege of Pharmacy, Gachon University, Incheon, Republic of KoreaCollege of Pharmacy, Gachon University, Incheon, Republic of KoreaCollege of Pharmacy, Gachon University, Incheon, Republic of KoreaGraduate School of Medicine, Korea University, Seoul, Republic of KoreaCollege of Pharmacy, Gachon University, Incheon, Republic of KoreaCollege of Pharmacy, Gachon University, Incheon, Republic of KoreaCollege of Pharmacy, Gachon University, Incheon, Republic of Korea; Corresponding author. College of Pharmacy, Gachon University, Incheon, Republic of KoreaCollege of Pharmacy, Gachon University, Incheon, Republic of Korea; Corresponding author. College of Pharmacy, Gachon University, Incheon, Republic of KoreaOxidative stress due to abnormal accumulation of reactive oxygen species (ROS) is an initiator of a large number of human diseases, and thus, the elimination and prevention of excessive ROS are important aspects of preventing the development of such diseases. Nuclear factor erythroid 2-related factor 2 (NRF2) is an essential transcription factor that defends against oxidative stress, and its function is negatively controlled by Kelch-like ECH-associated protein 1 (KEAP1). Therefore, activating NRF2 by inhibiting KEAP1 is viewed as a strategy for combating oxidative stress-related diseases. Here, we generated a cereblon (CRBN)-based proteolysis-targeting chimera (PROTAC), which we named SD2267, that induces the proteasomal degradation of KEAP1 and leads to NRF2 activation. As was intended, SD2267 bound to KEAP1, recruited CRBN, and induced the degradation of KEAP1. Furthermore, the KEAP1 degradation efficacy of SD2267 was diminished by MG132 (a proteasomal degradation inhibitor) but not by chloroquine (an autophagy inhibitor), which suggested that KEAP1 degradation by SD2267 was proteasomal degradation-dependent and autophagy-independent. Following KEAP1 degradation, SD2267 induced the nuclear translocation of NRF2, which led to the expression of NRF2 target genes and attenuated ROS accumulation induced by acetaminophen (APAP) in hepatocytes. Based on in vivo pharmacokinetic study, SD2267 was injected intraperitoneally at 1 or 3 mg/kg in APAP-induced liver injury mouse model. We observed that SD2267 degraded hepatic KEAP1 and attenuated APAP-induced liver damage. Summarizing, we described the synthesis of a KEAP1-targeting PROTAC (SD2267) and its efficacy and mode of action in vitro and in vivo. The results obtained suggest that SD2267 could be used to treat hepatic diseases related to oxidative stress.http://www.sciencedirect.com/science/article/pii/S2213231723001842Proteolysis-targeting chimera (PROTAC)KEAP1NRF2Oxidative stressLiver |
| spellingShingle | Se Yong Park Raju Gurung Jung Ho Hwang Ju-Hee Kang Hyun Jin Jung Alam Zeb Jong-Ik Hwang Sung Jean Park Han-Joo Maeng Dongyun Shin Seung Hyun Oh Development of KEAP1-targeting PROTAC and its antioxidant properties: In vitro and in vivo Redox Biology Proteolysis-targeting chimera (PROTAC) KEAP1 NRF2 Oxidative stress Liver |
| title | Development of KEAP1-targeting PROTAC and its antioxidant properties: In vitro and in vivo |
| title_full | Development of KEAP1-targeting PROTAC and its antioxidant properties: In vitro and in vivo |
| title_fullStr | Development of KEAP1-targeting PROTAC and its antioxidant properties: In vitro and in vivo |
| title_full_unstemmed | Development of KEAP1-targeting PROTAC and its antioxidant properties: In vitro and in vivo |
| title_short | Development of KEAP1-targeting PROTAC and its antioxidant properties: In vitro and in vivo |
| title_sort | development of keap1 targeting protac and its antioxidant properties in vitro and in vivo |
| topic | Proteolysis-targeting chimera (PROTAC) KEAP1 NRF2 Oxidative stress Liver |
| url | http://www.sciencedirect.com/science/article/pii/S2213231723001842 |
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