E3024 (3-but-2-ynyl-5-methyl-2-piperazin-1-yl-3,5-dihydro-4H-imidazo[4,5-d]pyridazin-4-one tosylate) is a dipeptidyl peptidase-IV (DPP-IV) inhibitor that was expected to be an antidiabetic agent. Its safety, tolerabil...E3024 (3-but-2-ynyl-5-methyl-2-piperazin-1-yl-3,5-dihydro-4H-imidazo[4,5-d]pyridazin-4-one tosylate) is a dipeptidyl peptidase-IV (DPP-IV) inhibitor that was expected to be an antidiabetic agent. Its safety, tolerability, pharmacokinetics (PK), and pharmacodynamics (PD) were investigated in a randomized, double-blind, placebo-controlled, ascending single-dose study in 48 healthy Japanese male subjects. Fasted subjects were orally administered E3024 (5, 10, 20, 40, or 80 mg) or placebo. E3024 was rapidly absorbed, with tmax values ranging between 0.33 and 3 h after dosing. The mean t1/2 ranged from 5.34 to 11.68 h. AUC0-inf and Cmax increased dose-proportionately. PK-PD relationship of E3024 was evaluated by using an Imax model, indicating that plasma E3024 concentrations and inhibitory effects of plasma DPP-IV activity were well correlated. The IC50 value was calculated as 33.7 ng/mL, which was consistent with in vitro data. Thus, E3024 showed a good PK profile and inhibited DPP-IV dose-dependently. Of 30 subjects administered E3024, 12 (40%) experienced adverse events (AEs). Dose escalation to 160 mg was abandoned owing to undesired subjective/objective findings in 4 of 6 subjects receiving 40 mg and 5 of 6 subjects receiving 80 mg. The most prominent AE was rash, but there were no serious AEs or deaths. The maximum tolerated dose was considered to be 20 mg. We hypothesized that histamine was a cause of the rash induction, and examined blood histamine levels of normal Fischer rats treated with E3024. Blood histamine levels were increased significantly by E3024 at 500 mg/kg (p < 0.001), but not by vildagliptin or valine-pyrrolidide (DPP-IV inhibitors) at the same dose. No blood histamine increases were observed in genetically mast cell-deficient Ws/Ws rats treated with E3024 at 500 mg/kg. In in vitro assays, E3024 induced histamine release from normal rat peritoneal mast cells in a concentration-dependent manner, but not from basophils. The structure-activity relationship study suggested that a piperazine group N-linked to the 2-position of the 5,6-membered fused heterocyclic rings was a key structural element for triggering histamine release.展开更多
In recent years,bioactive peptides targeting dipeptidyl peptidase-IV(DPP-IV)have emerged as promising candidates for managing type 2 diabetes.This study aimed to screen novel DPP-IV inhibitory(DPP-IVi)peptides from ra...In recent years,bioactive peptides targeting dipeptidyl peptidase-IV(DPP-IV)have emerged as promising candidates for managing type 2 diabetes.This study aimed to screen novel DPP-IV inhibitory(DPP-IVi)peptides from rabbit meat proteins using a combined approach of in silico study and in vitro experiments,with a focus on elucidating their molecular mechanisms.Initially,enzymatic simulation and virtual screening identified three potential DPP-IVi peptides.Among these,the tetrapeptide IPVK exhibited the strongest inhibitory activity(IC50=152.9±14.55μM),demonstrating competitive inhibition in vitro.Additionally,IPVK displayed robust resilience under diverse conditions,including variations in temperature,pH,and gastrointestinal digestion.Further analysis using molecular docking revealed IPVK’s interaction with critical residues within the S1,S2 pockets,and catalytic triad of DPP-IV through hydrogen bonds and hydrophobic interactions.Molecular dynamics simulations confirmed the stable formation of the IPVK-DPP-IV complex,providing insight into the peptide’s stability and binding affinity.Moreover,the highest occupied molecular orbital(HOMO)analysis highlighted isoleucine and proline as essential components for IPVK’s bioactivity.Supporting these findings,IPVK effectively inhibited native DPP-IV in Caco-2 cells(IC50=287.40±29.50μM)and exhibited hypoglycemic effects in vivo in mice.In summary,this study presents an efficient method for screening and identifying DPP-IVi peptides.The discovery of IPVK highlights rabbit meat proteins as a potential source of DPP-IV inhibitors,establishing a robust theoretical basis for its application in the management of type 2 diabetes.展开更多
This study investigated the DPP-IV inhibitory activity of taste-modulating peptides from porcine bone and elucidated their inhibitory mechanisms.Sensory evaluation identified seven peptides with effects on umami taste...This study investigated the DPP-IV inhibitory activity of taste-modulating peptides from porcine bone and elucidated their inhibitory mechanisms.Sensory evaluation identified seven peptides with effects on umami taste or saltiness enhancement.Computational tools predicted that they all possessed DPP-IV inhibitory potential.Post-digestion,GPTAANRM,GPGCKAGL,NLRDNYRF and GVNAMLRK exhibited superior DPP-IV inhibition(IC_(50):206.16-317.59μmol/L).Molecular docking revealed that all these four peptides interacted with active residues in S1,S2 and S3 pocket of DPP-IV and the Lineweaver-Burk plots indicated their competitive inhibition mechanism.All these peptides alleviated insulin resistance in HepG2 cells to some extent,with GPTAANRM and GPGCKAGL notably reducing the blood glucose curve area in C57BL/6 J mice.This research enhances our understanding of the beneficial role of taste-modulating peptides in diabetes management and suggests their potential in developing dietary strategies to optimize glycemic control in diabetic individuals.展开更多
Food-derived dipeptidyl peptidase IV(DPP-IV)inhibitors show promise as a therapeutic intervention for type 2 diabetic mellitus(T2DM).This investigation used a combination of virtual screening and enzymatic digestion t...Food-derived dipeptidyl peptidase IV(DPP-IV)inhibitors show promise as a therapeutic intervention for type 2 diabetic mellitus(T2DM).This investigation used a combination of virtual screening and enzymatic digestion to prepare and identify novel DPP-IV inhibitory peptides from Esox lucius,a typical cold-water fish from Xinjiang.Alkaline protease digest showed the best inhibitory effect of 62.04%±1.07%.The ultrafiltration U1(<3 kDa)fraction had the best inhibitory activity with an IC_(50) value of 2.77±0.09 mg/mL.From the dextran gel chromatography S2 fraction,543 peptide fragments were found,and six peptides underwent virtual screening.Among them,APDPF showed promising DPP-IV inhibitory activity with an IC_(50) value of 0.38±0.02 mg/mL and exhibited competitive inhibition of DPP-IV by binding to amino acid residues in the S1 and S2 active pockets by forming hydrogen bonds and hydrophobic forces.This study provides a new way to prepare DPP-IV inhibitory peptide and is of great theoretical significance and practical value for further exploring the development of endogenous peptides from Esox lucius.展开更多
The objective of this study was to isolate and purify DPP-IV inhibitory(DPP-IVi)peptides from rabbit meat hydrolysate(RMH)and explore their mechanism of action,stability,and hypoglycemic effects.Various pro-teases,inc...The objective of this study was to isolate and purify DPP-IV inhibitory(DPP-IVi)peptides from rabbit meat hydrolysate(RMH)and explore their mechanism of action,stability,and hypoglycemic effects.Various pro-teases,including pepsin,alcalase,compound proteinase,flavourzyme,and bromelain,were utilized to prepare RMH,with RMH derived from the compound proteinase demonstrating the highest DPP-IV inhibition activity.Employing gel filtration chromatography,RP-HPLC and UPLC-MS/MS,we identified Leucyl-Leucine(LL),a dipeptide exhibiting potent DPP-IV inhibitory activity(IC_(50)=99.85±5.45μM).LL exhibited resistance to simulated gastrointestinal digestion in vitro and competitively inhibited DPP-IV.Molecular docking analysis revealed LL’s formation of hydrogen bonds and hydrophobic interactions with key amino acid residues of DPP-IV.Furthermore,LL exhibits good inhibitory activity in situ against DPP-IV.,with an IC_(50) value of 207.40±21.50μM.In an oral glucose tolerance test(OGTT),LL displayed significant hypoglycemic effects at 30 min after administration.In conclusion,LL derived from rabbit meat hydrolysate exhibited remarkable DPP-IV inhibition activity and hypoglycemic effects,suggesting its potential as a therapeutic candidate for diabetes treatment.展开更多
通过抑制二肽基肽酶IV(Dipeptidyl-peptidase IV, DPP-IV)的活性,从而减少GLP-1的降解,提高血液中胰岛素的水平,是控制血糖水平的一种重要手段。本文以海参(Holothariatubulosa)为原料,通过探究蛋白酶种类、加酶量和酶解时间对酶解产物D...通过抑制二肽基肽酶IV(Dipeptidyl-peptidase IV, DPP-IV)的活性,从而减少GLP-1的降解,提高血液中胰岛素的水平,是控制血糖水平的一种重要手段。本文以海参(Holothariatubulosa)为原料,通过探究蛋白酶种类、加酶量和酶解时间对酶解产物DPP-IV抑制活性、蛋白回收率和水解度的影响,确定了海参DPP-IV抑制肽的制备条件,并进一步测定了其分子量分布和总氨基酸组成,最后通过UPLC-MS/MS鉴定了其潜在的活性肽序列。结果发现,木瓜蛋白酶与复合蛋白酶1:1的复配酶解具有最佳的酶解效果,其产物的得率和DPP-IV抑制活性均最高,且在加酶量为干海参质量的1%,酶解时间为4 h时,海参酶解产物在终浓度为2 mg/mL时的DPP-IV抑制率为66.97%,蛋白回收率为76.25%,水解度为6.10%。酶解产物的分子量大都小于5000 u,且富含脯氨酸(Pro)和丙氨酸(Ala)等与抑制DPP-IV活性有关的氨基酸。将获得的酶解物中的肽段进行液质联用检测,并通过Mascot分析筛选得到28条具DPP-IV抑制肽的肽序列,分子量在500~1936 u。本实验结果为以海参为原料进行降血糖产品的开发奠定了基础。展开更多
The use of green and low-cost techniques to produce bioactive peptides with anti-diabetic properties from protein-rich fish waste is a sustainable way to manage agro-fishery and food industrial waste.This study invest...The use of green and low-cost techniques to produce bioactive peptides with anti-diabetic properties from protein-rich fish waste is a sustainable way to manage agro-fishery and food industrial waste.This study investigated a novel ultrasound(UL)assisted deep eutectic solvent(DES)based extraction approach to recover bioactive protein-rich extracts from tilapia fish viscera waste under optimized conditions(52.52%ultrasound amplitude,1:4 DES molar ratio,25 min extraction time).The aqueous two-phase system(ATPS)method suc-cessfully concentrated the crude proteins in the top phase and back-extracted proteins to the bottom phase with a high recovery yield.Hydrolysis by Alcalase produced tilapia viscera protein hydrolysate(TVPH)with low mo-lecular weight(MW)peptides and low dipeptidyl peptidase(DPP)-IV half-maximal inhibitory concentration(IC_(50))value of 1.57±0.16 mg/mL.Notably,TVPH-3 fraction and TVPH-3-2 sub-fraction exhibited appreciable DPP-IV IC_(50) values of 1.36±0.08 mg/mL and 0.42±0.01 mg/mL,respectively.Ultrafiltration combined with RP-HPLC enriched the DPP-IV inhibitory concentration of the individual protein fractions.TVPH-3-2 sub-fraction contained hydrophobic amino acids and peptide sequences with typical DPP-IV inhibitory peptide sequence motifs.The results presented in this study indicate that tilapia viscera biomass is a potential source for protein recovery aided with a green extraction approach,and tilapia viscera protein-derived hydrolysates have potential applications as a potent functional food ingredient for the management of metabolic diseases like type-2 diabetes.展开更多
文摘E3024 (3-but-2-ynyl-5-methyl-2-piperazin-1-yl-3,5-dihydro-4H-imidazo[4,5-d]pyridazin-4-one tosylate) is a dipeptidyl peptidase-IV (DPP-IV) inhibitor that was expected to be an antidiabetic agent. Its safety, tolerability, pharmacokinetics (PK), and pharmacodynamics (PD) were investigated in a randomized, double-blind, placebo-controlled, ascending single-dose study in 48 healthy Japanese male subjects. Fasted subjects were orally administered E3024 (5, 10, 20, 40, or 80 mg) or placebo. E3024 was rapidly absorbed, with tmax values ranging between 0.33 and 3 h after dosing. The mean t1/2 ranged from 5.34 to 11.68 h. AUC0-inf and Cmax increased dose-proportionately. PK-PD relationship of E3024 was evaluated by using an Imax model, indicating that plasma E3024 concentrations and inhibitory effects of plasma DPP-IV activity were well correlated. The IC50 value was calculated as 33.7 ng/mL, which was consistent with in vitro data. Thus, E3024 showed a good PK profile and inhibited DPP-IV dose-dependently. Of 30 subjects administered E3024, 12 (40%) experienced adverse events (AEs). Dose escalation to 160 mg was abandoned owing to undesired subjective/objective findings in 4 of 6 subjects receiving 40 mg and 5 of 6 subjects receiving 80 mg. The most prominent AE was rash, but there were no serious AEs or deaths. The maximum tolerated dose was considered to be 20 mg. We hypothesized that histamine was a cause of the rash induction, and examined blood histamine levels of normal Fischer rats treated with E3024. Blood histamine levels were increased significantly by E3024 at 500 mg/kg (p < 0.001), but not by vildagliptin or valine-pyrrolidide (DPP-IV inhibitors) at the same dose. No blood histamine increases were observed in genetically mast cell-deficient Ws/Ws rats treated with E3024 at 500 mg/kg. In in vitro assays, E3024 induced histamine release from normal rat peritoneal mast cells in a concentration-dependent manner, but not from basophils. The structure-activity relationship study suggested that a piperazine group N-linked to the 2-position of the 5,6-membered fused heterocyclic rings was a key structural element for triggering histamine release.
基金supported by Chongqing Forestry Reform and Development Fund(grant number Yu-lin-ke-tui-2019-2)Chongqing Normal University Fund projects(grant number 19XLB006).
文摘In recent years,bioactive peptides targeting dipeptidyl peptidase-IV(DPP-IV)have emerged as promising candidates for managing type 2 diabetes.This study aimed to screen novel DPP-IV inhibitory(DPP-IVi)peptides from rabbit meat proteins using a combined approach of in silico study and in vitro experiments,with a focus on elucidating their molecular mechanisms.Initially,enzymatic simulation and virtual screening identified three potential DPP-IVi peptides.Among these,the tetrapeptide IPVK exhibited the strongest inhibitory activity(IC50=152.9±14.55μM),demonstrating competitive inhibition in vitro.Additionally,IPVK displayed robust resilience under diverse conditions,including variations in temperature,pH,and gastrointestinal digestion.Further analysis using molecular docking revealed IPVK’s interaction with critical residues within the S1,S2 pockets,and catalytic triad of DPP-IV through hydrogen bonds and hydrophobic interactions.Molecular dynamics simulations confirmed the stable formation of the IPVK-DPP-IV complex,providing insight into the peptide’s stability and binding affinity.Moreover,the highest occupied molecular orbital(HOMO)analysis highlighted isoleucine and proline as essential components for IPVK’s bioactivity.Supporting these findings,IPVK effectively inhibited native DPP-IV in Caco-2 cells(IC50=287.40±29.50μM)and exhibited hypoglycemic effects in vivo in mice.In summary,this study presents an efficient method for screening and identifying DPP-IVi peptides.The discovery of IPVK highlights rabbit meat proteins as a potential source of DPP-IV inhibitors,establishing a robust theoretical basis for its application in the management of type 2 diabetes.
基金supported by the National Natural Science Foundation of China(Nos.32402278 and 32372463).
文摘This study investigated the DPP-IV inhibitory activity of taste-modulating peptides from porcine bone and elucidated their inhibitory mechanisms.Sensory evaluation identified seven peptides with effects on umami taste or saltiness enhancement.Computational tools predicted that they all possessed DPP-IV inhibitory potential.Post-digestion,GPTAANRM,GPGCKAGL,NLRDNYRF and GVNAMLRK exhibited superior DPP-IV inhibition(IC_(50):206.16-317.59μmol/L).Molecular docking revealed that all these four peptides interacted with active residues in S1,S2 and S3 pocket of DPP-IV and the Lineweaver-Burk plots indicated their competitive inhibition mechanism.All these peptides alleviated insulin resistance in HepG2 cells to some extent,with GPTAANRM and GPGCKAGL notably reducing the blood glucose curve area in C57BL/6 J mice.This research enhances our understanding of the beneficial role of taste-modulating peptides in diabetes management and suggests their potential in developing dietary strategies to optimize glycemic control in diabetic individuals.
基金funded by XPCC Key areas Science and Technology Program of China(grant number 2023AB006-04)XPCC Guiding Science and Technology Program of China(grant number 2022DZ016)+2 种基金High-level Talent Initiation Program of Shihezi University(grant number RCZK202355)Key Laboratory for Nutrition and Safety of Xinjiang Specialty Foods,Shihezi City of Eighth Division(grant number 2022PT02)Research Program of Shihezi University(grant number PYZK202405).
文摘Food-derived dipeptidyl peptidase IV(DPP-IV)inhibitors show promise as a therapeutic intervention for type 2 diabetic mellitus(T2DM).This investigation used a combination of virtual screening and enzymatic digestion to prepare and identify novel DPP-IV inhibitory peptides from Esox lucius,a typical cold-water fish from Xinjiang.Alkaline protease digest showed the best inhibitory effect of 62.04%±1.07%.The ultrafiltration U1(<3 kDa)fraction had the best inhibitory activity with an IC_(50) value of 2.77±0.09 mg/mL.From the dextran gel chromatography S2 fraction,543 peptide fragments were found,and six peptides underwent virtual screening.Among them,APDPF showed promising DPP-IV inhibitory activity with an IC_(50) value of 0.38±0.02 mg/mL and exhibited competitive inhibition of DPP-IV by binding to amino acid residues in the S1 and S2 active pockets by forming hydrogen bonds and hydrophobic forces.This study provides a new way to prepare DPP-IV inhibitory peptide and is of great theoretical significance and practical value for further exploring the development of endogenous peptides from Esox lucius.
基金supported by Chongqing Forestry Reform and Development Fund(grant number Yu-lin-ke-tui-2019-2)the Project of Chongqing Science and Technology Bureau(grant number cstc2017chmsxdny0246).
文摘The objective of this study was to isolate and purify DPP-IV inhibitory(DPP-IVi)peptides from rabbit meat hydrolysate(RMH)and explore their mechanism of action,stability,and hypoglycemic effects.Various pro-teases,including pepsin,alcalase,compound proteinase,flavourzyme,and bromelain,were utilized to prepare RMH,with RMH derived from the compound proteinase demonstrating the highest DPP-IV inhibition activity.Employing gel filtration chromatography,RP-HPLC and UPLC-MS/MS,we identified Leucyl-Leucine(LL),a dipeptide exhibiting potent DPP-IV inhibitory activity(IC_(50)=99.85±5.45μM).LL exhibited resistance to simulated gastrointestinal digestion in vitro and competitively inhibited DPP-IV.Molecular docking analysis revealed LL’s formation of hydrogen bonds and hydrophobic interactions with key amino acid residues of DPP-IV.Furthermore,LL exhibits good inhibitory activity in situ against DPP-IV.,with an IC_(50) value of 207.40±21.50μM.In an oral glucose tolerance test(OGTT),LL displayed significant hypoglycemic effects at 30 min after administration.In conclusion,LL derived from rabbit meat hydrolysate exhibited remarkable DPP-IV inhibition activity and hypoglycemic effects,suggesting its potential as a therapeutic candidate for diabetes treatment.
基金the National Kaohsiung University of Science and Technology,Kaohsiung City,Taiwan for technical supportsupported by the National Science and Technology Council(NSTC)project,Taiwan(Ref.No.112-2222-E-992-005)Cheng-Di Dong also acknowledges the Taiwan MOST for funding support(Ref.No.109-2222-E-992-002).
文摘The use of green and low-cost techniques to produce bioactive peptides with anti-diabetic properties from protein-rich fish waste is a sustainable way to manage agro-fishery and food industrial waste.This study investigated a novel ultrasound(UL)assisted deep eutectic solvent(DES)based extraction approach to recover bioactive protein-rich extracts from tilapia fish viscera waste under optimized conditions(52.52%ultrasound amplitude,1:4 DES molar ratio,25 min extraction time).The aqueous two-phase system(ATPS)method suc-cessfully concentrated the crude proteins in the top phase and back-extracted proteins to the bottom phase with a high recovery yield.Hydrolysis by Alcalase produced tilapia viscera protein hydrolysate(TVPH)with low mo-lecular weight(MW)peptides and low dipeptidyl peptidase(DPP)-IV half-maximal inhibitory concentration(IC_(50))value of 1.57±0.16 mg/mL.Notably,TVPH-3 fraction and TVPH-3-2 sub-fraction exhibited appreciable DPP-IV IC_(50) values of 1.36±0.08 mg/mL and 0.42±0.01 mg/mL,respectively.Ultrafiltration combined with RP-HPLC enriched the DPP-IV inhibitory concentration of the individual protein fractions.TVPH-3-2 sub-fraction contained hydrophobic amino acids and peptide sequences with typical DPP-IV inhibitory peptide sequence motifs.The results presented in this study indicate that tilapia viscera biomass is a potential source for protein recovery aided with a green extraction approach,and tilapia viscera protein-derived hydrolysates have potential applications as a potent functional food ingredient for the management of metabolic diseases like type-2 diabetes.
