Human cytochrome P4501B1(h CYP1B1),an extrahepatic heme-dependent monooxygenase,has been validated as a key target for overcoming chemotherapy resistance and tumorigenesis.Herein,to discover novel efficacious h CYP1B1...Human cytochrome P4501B1(h CYP1B1),an extrahepatic heme-dependent monooxygenase,has been validated as a key target for overcoming chemotherapy resistance and tumorigenesis.Herein,to discover novel efficacious h CYP1B1 inhibitors,a suite of 1,8-naphthalimide derivatives was designed,synthesized,and biologically evaluated,via integrating structure-based drug design(SBDD)and biochemical assays.After two rounds of structural modifications and structure-activity relationship(SAR)studies,the results suggested that introducing a benzene ring at the north part and a halogen atom at the C-4 site significantly enhanced the anti-h CYP1B1 effects of naphthalimides.Among all tested 1,8-naphthalimides,NB-10showed the most potent anti-h CYP1B1 effect(half maximal inhibitory concentration(IC_(50))=0.41 nmol/L)and excellent specificity,while this agent did not activate Ah R transcription activity in living cells.Further cellular assays and in vivo tests in paclitaxel(PTX)-resistance xenograft mice showed that NB-10could significantly potentiate the anti-cancer effects of PTX both in vitro and in vivo,while this agent also showed high safety profiles in mice.Mechanistically,NB-10 potently inhibited h CYP1B1-catalyzed 7-ethoxyresorufin O-deethylation in a competitive manner,with an estimated Kivalue of 0.15 nmol/L.Docking simulations showed that NB-10 could be well-fitted in the catalytic pocket of h CYP1B1 to form a stable conformation with a high binding affinity.Collectively,several potent 4-halogenated naphthalimides were developed as novel h CYP1B1 inhibitors,while NB-10 showed high safety profiles and impressive efficacy for overcoming h CYP1B1-associated PTX resistance both in vitro and in vivo.展开更多
Objective:Qingfei oral liquid(QF),an experimental Chinese medicine prescription developed from the ancient priscription of traditional Chinese medicines Ma Xin Shi Gan decoction and Tingli Dazao Xie Fei decoction,has ...Objective:Qingfei oral liquid(QF),an experimental Chinese medicine prescription developed from the ancient priscription of traditional Chinese medicines Ma Xin Shi Gan decoction and Tingli Dazao Xie Fei decoction,has been effectively used since decades to treat patients with viral pneumonia and asthma.In our previous study,we had demonstrated that QF can significantly reduce airway hyperresponsiveness,hyperemia,lung tissue edema,inflammatory lung tissue infiltration in mice,airway mucus secretion,and peripheral airway collagen hyperplasia;however,its mechanism of action is unknown.Methods:Fifty 6–8-week-old male BALB/c mice were equally and randomly divided into five groups:the control,ovalbumin(OVA),OVA+respiratory syncytial virus(RSV),QF,and dexamethasone(Dxms)groups.The QF group was administered QF at 1.17 g·kg−1·d−1,the Dxms group received dexamethasone injections at 0.2 mg·kg−1·d−1,and the remaining groups were administered PBS.Inflammation in the lung tissue was assessed by hematoxylin and eosin(HE),periodic acid–Schiff(PAS),and Van Gieson staining.ELISA was used to evaluate the IL-13,IL-25,and IL-33 in the mice.Western blotting was used to examine changes in the proteins levels of transient receptor potential vanilloid-1(TRPV1)and mucin 5AC(MUC5AC)in the lung tissues of mice.Results:Histopathological evaluation revealed that the OVA and OVA+RSV groups exhibited lung tissue edema and inflammatory lung tissue infiltration in the HE staining and airway secretions in the PAS staining;collagen hyperplasia around the airway was increased in these two groups compared with the control group.The QF group exhibited significantly reduced lung tissue edema,inflammatory lung tissue infiltration,airway secretions,and collagen hyperplasia around the airway compared with the OVA+RSV group.We analyzed the serum levels of IL-13,IL-25,and IL-33 in the mice and found that these levels were higher in the OVA and OVA+RSV groups than in the control group(P<0.05 in the OVA group,P<0.01 in the OVA+RSV group).The QF group exhibited significantly decreased serum levels of IL-13,IL-25,and IL-33 compared with the OVA+RSV group(all P<0.05).The Dxms group also exhibited significant decreases in the serum levels of IL-13 and IL-33(all P<0.05)but no significant decrease in the serum levels of IL-25 compared with the RSV+OVA group.Finally,we examined the protein levels of TRPV1 and MUC5AC in the lung tissues of mice using Western blotting.After identifying RSV infection in the mice with asthma,the protein levels of TRPV1 and MUC5AC in the lung tissues of mice were significantly higher than those in the control group(P<0.05,P<0.01).We found that compared with RSV+OVA,QF can significantly downregulate the protein level of TRPV1;further,the protein level of MUC5AC was also significantly reduced(all P<0.001).Conclusion:QF can inhibit RSV replication and reduce airway inflammation and mucus hypersecretion injury caused by RSV infection and asthma,and its mechanism of action may be associated with the downregulation of TRPV1 expression and a decrease in airway mucus hypersecretion injury.展开更多
基金supported by the National Natural Science Foundation of China(Nos.82273897,U23A20516,32101202)Organizational Key Research and Development Program of Shanghai University of Traditional Chinese Medicine(No.2023YZZ02)+5 种基金Shanghai Municipal Health Commission’s TCM research project(No.2022CX005)Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(No.ZYYCXTDD-202004)Pudong Institute of Clinical Chinese Medicine(No.YC-2023-0603)The“Fourteenth Five-Year Plan”Traditional Chinese Medicine Specialty Project for the Construction of Andrology Departments in TCM(No.ZYTSZK1-4)the State Key Laboratory of Fine Chemicals,Dalian University of Technology(No.KF2202)the Fundamental Research Funds for the Central Universities(No.G2024KY05106)。
文摘Human cytochrome P4501B1(h CYP1B1),an extrahepatic heme-dependent monooxygenase,has been validated as a key target for overcoming chemotherapy resistance and tumorigenesis.Herein,to discover novel efficacious h CYP1B1 inhibitors,a suite of 1,8-naphthalimide derivatives was designed,synthesized,and biologically evaluated,via integrating structure-based drug design(SBDD)and biochemical assays.After two rounds of structural modifications and structure-activity relationship(SAR)studies,the results suggested that introducing a benzene ring at the north part and a halogen atom at the C-4 site significantly enhanced the anti-h CYP1B1 effects of naphthalimides.Among all tested 1,8-naphthalimides,NB-10showed the most potent anti-h CYP1B1 effect(half maximal inhibitory concentration(IC_(50))=0.41 nmol/L)and excellent specificity,while this agent did not activate Ah R transcription activity in living cells.Further cellular assays and in vivo tests in paclitaxel(PTX)-resistance xenograft mice showed that NB-10could significantly potentiate the anti-cancer effects of PTX both in vitro and in vivo,while this agent also showed high safety profiles in mice.Mechanistically,NB-10 potently inhibited h CYP1B1-catalyzed 7-ethoxyresorufin O-deethylation in a competitive manner,with an estimated Kivalue of 0.15 nmol/L.Docking simulations showed that NB-10 could be well-fitted in the catalytic pocket of h CYP1B1 to form a stable conformation with a high binding affinity.Collectively,several potent 4-halogenated naphthalimides were developed as novel h CYP1B1 inhibitors,while NB-10 showed high safety profiles and impressive efficacy for overcoming h CYP1B1-associated PTX resistance both in vitro and in vivo.
基金This work was supported by Natural Science Foundation of China(81674020).
文摘Objective:Qingfei oral liquid(QF),an experimental Chinese medicine prescription developed from the ancient priscription of traditional Chinese medicines Ma Xin Shi Gan decoction and Tingli Dazao Xie Fei decoction,has been effectively used since decades to treat patients with viral pneumonia and asthma.In our previous study,we had demonstrated that QF can significantly reduce airway hyperresponsiveness,hyperemia,lung tissue edema,inflammatory lung tissue infiltration in mice,airway mucus secretion,and peripheral airway collagen hyperplasia;however,its mechanism of action is unknown.Methods:Fifty 6–8-week-old male BALB/c mice were equally and randomly divided into five groups:the control,ovalbumin(OVA),OVA+respiratory syncytial virus(RSV),QF,and dexamethasone(Dxms)groups.The QF group was administered QF at 1.17 g·kg−1·d−1,the Dxms group received dexamethasone injections at 0.2 mg·kg−1·d−1,and the remaining groups were administered PBS.Inflammation in the lung tissue was assessed by hematoxylin and eosin(HE),periodic acid–Schiff(PAS),and Van Gieson staining.ELISA was used to evaluate the IL-13,IL-25,and IL-33 in the mice.Western blotting was used to examine changes in the proteins levels of transient receptor potential vanilloid-1(TRPV1)and mucin 5AC(MUC5AC)in the lung tissues of mice.Results:Histopathological evaluation revealed that the OVA and OVA+RSV groups exhibited lung tissue edema and inflammatory lung tissue infiltration in the HE staining and airway secretions in the PAS staining;collagen hyperplasia around the airway was increased in these two groups compared with the control group.The QF group exhibited significantly reduced lung tissue edema,inflammatory lung tissue infiltration,airway secretions,and collagen hyperplasia around the airway compared with the OVA+RSV group.We analyzed the serum levels of IL-13,IL-25,and IL-33 in the mice and found that these levels were higher in the OVA and OVA+RSV groups than in the control group(P<0.05 in the OVA group,P<0.01 in the OVA+RSV group).The QF group exhibited significantly decreased serum levels of IL-13,IL-25,and IL-33 compared with the OVA+RSV group(all P<0.05).The Dxms group also exhibited significant decreases in the serum levels of IL-13 and IL-33(all P<0.05)but no significant decrease in the serum levels of IL-25 compared with the RSV+OVA group.Finally,we examined the protein levels of TRPV1 and MUC5AC in the lung tissues of mice using Western blotting.After identifying RSV infection in the mice with asthma,the protein levels of TRPV1 and MUC5AC in the lung tissues of mice were significantly higher than those in the control group(P<0.05,P<0.01).We found that compared with RSV+OVA,QF can significantly downregulate the protein level of TRPV1;further,the protein level of MUC5AC was also significantly reduced(all P<0.001).Conclusion:QF can inhibit RSV replication and reduce airway inflammation and mucus hypersecretion injury caused by RSV infection and asthma,and its mechanism of action may be associated with the downregulation of TRPV1 expression and a decrease in airway mucus hypersecretion injury.
