Molecules最新文献

{"title":"Simple Method for Controlling Gold Nanocluster Size in Mesoporous Silica: SBA-11.","authors":"Tariq Aqeel, Ali Bumajdad","doi":"10.3390/molecules30092035","DOIUrl":"10.3390/molecules30092035","url":null,"abstract":"<p><p>Mesoporous silica containing Au nanoclusters has been widely used in various fields owing to its desirable properties and functionality. This work introduces a facile method to control the size of Au nanoclusters within silica hosts. This was achieved by applying a reducing gas at a controlled temperature to obtain a narrow or broad Au cluster size distribution inside the silica host. The Au nanoclusters and their silica hosts were characterized through X-ray diffraction, high-resolution transmission electron microscopy, energy-dispersive X-ray spectroscopy, ultraviolet-visible spectroscopy, N₂ sorption analysis, and X-ray photoelectron spectroscopy. The average size of the Au nanoclusters was 1.3 nm in the narrow-distribution sample. In comparison, the wide-distribution sample exhibited two cluster populations: 1-2 nm inside the pores and 5-30 nm outside the pores. Methylene blue (M.B.) photocatalysis was conducted under direct sunlight to evaluate the catalytic activity of these materials. Sample 1-Au-SBA-11, containing Au clusters averaging 1.3 nm, achieved substantial M.B. degradation within 90 min-half the time required by the 2-Au-SBA-11 sample. Such Au clusters of different size ranges have potential applications across diverse fields.</p>","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12073877/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144002447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nutritional and Functional Values of Grape Seed Flour and Extract for Production of Antioxidative Dietary Supplements and Functional Foods.","authors":"Yavor Ivanov, Milka Atanasova, Tzonka Godjevargova","doi":"10.3390/molecules30092029","DOIUrl":"10.3390/molecules30092029","url":null,"abstract":"<p><p>The potential of the seed flours and extracts of the red grapes Pinot Noir and Marselan for application as food additives and antioxidative dietary supplements was determined. The differences between the quality characteristics of the seeds, flour, and extracts of the two grape varieties were examined. The polyphenol composition and antioxidant potential of the two extracts were compared. The extracts were rich in polyphenols, especially flavonoids (52.01 mg QE/g DW) and procyanidins (152.18 mg CE/g DW). The nutritional composition of the flours and extracts was determined. It was found that the content of ash, crude protein, crude fat, and total dietary fibers in the flour from the studied grape seeds was about 1.5 times higher than that in the extracts. Only the carbohydrate content was 2-2.5 times higher in both extracts compared to that in the flours. The content of macro and microelements in the seed extracts and in the flours was high. The inhibitory potential of the two seed extracts on three key enzymes affecting diabetes and obesity-α-amylase, α-glucosidase, and pancreatic lipase-was studied. The highest degree of extract inhibition against α-glucosidase was determined (IC₅₀-2.53 ± 0.24 µg/mL). A real inhibitory assessment of the extracts was made by implementing an in vitro digestion simulation method. It was found that the percentage of inhibition of the enzymes with the digested extract was higher compared to those with the undigested extract in buffer and salt solution. Our study proves that the high content of flavonoids and procyanidins in the two extracts determines their high inhibitory capacity against the three enzymes and their potential for managing diabetes and obesity.</p>","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12073665/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144002622","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis and Biological Evaluation of Novel Ramalin Derivatives as Multi-Target Agents for Alzheimer's Disease.","authors":"Tai Kyoung Kim, Ju-Mi Hong, Yongeun Cho, Yeji Jeon, Heewon Cho, Jeongmi Lee, Jaewon Kim, Kyung Hee Kim, Il-Chan Kim, Se Jong Han, Hyuncheol Oh, Dong-Gyu Jo, Joung Han Yim","doi":"10.3390/molecules30092030","DOIUrl":"10.3390/molecules30092030","url":null,"abstract":"<p><p>Alzheimer's disease (AD) is a complex neurodegenerative disorder characterized by cognitive decline, oxidative stress, neuroinflammation, amyloid-beta (Aβ) accumulation, and tau protein hyperphosphorylation. In this study, we synthesized novel Ramalin derivatives and evaluated their therapeutic potential against AD, focusing on antioxidant, anti-inflammatory, and neuroprotective activities. RA-2OMe, RA-4OMe, RA-2CF3, and RA-4OCF3 showed strong antioxidant effects, while RA-2OMe exhibited potent NO and NLRP3 inhibition (~20%). RA-NAP, RA-PYD, and RA-2Q showed moderate anti-inflammatory activity. BACE-1 inhibition was significant in RA-3CF3, RA-NAP, and RA-PYD, with IC₅₀ values lower than that of positive control, indicating greater inhibitory potency. RA-NAP and RA-PYD effectively inhibited both Aβ and tau aggregation, highlighting their multi-target potential for AD therapy. These findings indicate that Ramalin derivatives exhibit potential for multi-target activity in AD treatment. However, further studies on their pharmacokinetics, in vivo efficacy, and long-term safety are required to confirm their therapeutic applicability.</p>","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12073177/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144005551","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preparation and Electrochemical Characteristics of the Co-Doped Li₇La₃Zr₂O₁₂ Solid Electrolyte with Fe³⁺ and Bi³.","authors":"Jialu Qu, Xingyu Duan, Ke Xue, Shengli An","doi":"10.3390/molecules30092028","DOIUrl":"10.3390/molecules30092028","url":null,"abstract":"<p><p>Solid-state electrolytes (SSEs) have emerged as the most promising alternative to liquid electrolytes in batteries due to their enhanced stability and safety. Among these, the garnet-type Li₇La₃Zr₂O₁₂ (LLZO) solid electrolyte has attracted significant research interest due to its wide electrochemical stability window and good air stability. However, the ionic conductivity of LLZO is lower due to its high sintering temperature and unstable phase structure. In this study, Li_6.4+xFe_0.2La₃Zr_2-xBi_xO₁₂ (x = 0, 0.05, 0.1, 0.15) solid electrolytes were synthesized using a conventional solid-state reaction method by co-doping LLZO with Fe³⁺ and Bi³⁺ ions. Compared with pure LLZO, doping with Fe³⁺ effectively stabilizes the cubic phase, thereby enhancing the ionic conductivity. Moreover, Bi³⁺ doping significantly lowers the sintering temperature of the electrolyte, which in turn reduces energy consumption during the processing. The co-doping of Fe³⁺ and Bi³⁺ not only improves the density of the LLZO electrolyte, achieving a relative density of up to 95%, but also increases the ionic conductivity, with a maximum value of 7.57 × 10^-4 S·cm^-1 observed at the optimal composition (Li_6.4+xFe_0.2La₃Zr_2-xBi_xO₁₂, x = 0.1).</p>","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12073242/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144032149","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Influence of Electron Transfer Mediators in the Pd(II)-Catalyzed Oxidative Carbonylation of Aniline.","authors":"Andrea Vavasori, Lucio Ronchin, Luca Pietrobon, Sara Bravo","doi":"10.3390/molecules30092027","DOIUrl":"10.3390/molecules30092027","url":null,"abstract":"<p><p>Currently, the most promising alternative to the use of the phosgenation reaction, for large-scale production of isocyanates, ureas, and carbamates, appears to be the Pd-catalyzed oxidative carbonylation of arylamines. During the reaction, the Pd(II) catalytic species are reduced to Pd(0) and the addition of sacrificial oxidizing agents is usually necessary to restart the catalytic cycle. Among these oxidizing agents, molecular oxygen is undoubtedly the more appealing, from an economical and green point of view, but it is not so efficient, whereas several metal salts (named cocatalysts) can be used, able to form redox couples with Pd(0) or to act as electron transfer mediators with oxygen itself. Testing several Pd(II) complexes, metal cocatalysts, and promoters, we have found that the [PdCl₂(dppf)]/FeCl₃/LiBr = 1/1200/200 (mol/mol) system efficiently catalyzes the carbonylation of aniline to form 1,3-diphenylurea selectively (100%) with a TOF of ca. 1177 h^-1. On the other hand, the addition of oxygen to such a system strongly increases the aniline conversion (0.3 MPa of O₂ increases the TOF at ca. 3930 h^-1), but it moves the selectivity towards the phenyl isocyanate (65%, mol/mol).</p>","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12074507/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144011733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PF-06447475 Molecule Attenuates the Neuropathology of Familial Alzheimer's and Coexistent Parkinson's Disease Markers in PSEN1 I416T Dopaminergic-like Neurons.","authors":"Diana Alejandra Quintero-Espinosa, Carlos Velez-Pardo, Marlene Jimenez-Del-Rio","doi":"10.3390/molecules30092034","DOIUrl":"10.3390/molecules30092034","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Familial Alzheimer's disease (FAD) is a complex multifactorial disorder clinically characterized by cognitive impairment and memory loss. Pathologically, FAD is characterized by intracellular accumulation of the protein fragment Aβ42 (iAβ), hyperphosphorylated microtubule-associated protein TAU (p-TAU), and extensive degeneration of basal forebrain cholinergic neurons of the nucleus basalis of Meynert (NbM) and the medial septal nucleus (MSN), mainly caused by mutations in the amyloid precursor protein (&lt;i&gt;APP),&lt;/i&gt; presenilin 1 (&lt;i&gt;PSEN1&lt;/i&gt;), and &lt;i&gt;PSEN2&lt;/i&gt; gene. Since the dopaminergic system may contribute to FAD symptoms, alterations in the nigro-hippocampal pathway may be associated with cognitive impairment in FAD. Interestingly, p-α-synuclein (p-α-Syn), Aβ, and p-TAU have been found to coexist in vulnerable regions of postmortem AD brains. However, the mechanism by which Aβ, p-TAU, and α-Syn coexist in DAergic neurons in AD brains has not been determined. We generated PSEN1 I416T dopaminergic-like neurons (DALNs) from I416T menstrual stromal cells (MenSCs) in &lt;i&gt;NeuroForsk 2.0&lt;/i&gt; medium for 7 days and then cultured them in minimal culture medium (MCm) for another 4 days. On day 11, DALNs were analyzed for molecular and pathological markers by flow cytometry and fluorescence microscopy. We found that mutant DALNs showed increased accumulation of iAβ as well as increased phosphorylation of TAU at S202/T205 compared to WT DALNs. Thus, mutant DALNs exhibited typical pathological hallmarks of Alzheimer's disease. Furthermore, PSEN1 I416T DALNs showed concomitant signs of OS as evidenced by the appearance of oxidized sensor protein DJ-1 (i.e., DJ-1C106-SO&lt;sub&gt;3&lt;/sub&gt;) and apoptotic markers TP53, pS63-c-JUN, PUMA, and cleavage caspase 3 (CC3). Notably, these DALNs exhibited PD-associated proteins such as intracellular accumulation of α-Syn (detected as aggregates of pS129-α-Syn) and phosphorylation of LRRK2 kinase at residue S935. In addition, mutant DALNs showed a 17.16- and 6.17-fold decrease in DA-induced Ca&lt;sup&gt;2+&lt;/sup&gt; flux, compared to WT DALNs. These observations suggest that iAβ and p-TAU, together with p-α-Syn, and p-LRRK2 kinase, may damage DAergic neurons and thereby contribute to the exacerbation of neuropathologic processes in FAD. Remarkably, the LRRK2 inhibitor PF-06447475 (PF-475) significantly reversed PSEN1 I416T-induced neuropathological markers in DAergic neurons. PF-465 inhibitor reduced iAβ, oxDJ-1C106-SO&lt;sub&gt;3&lt;/sub&gt;, and p-TAU. In addition, this inhibitor reduced pS935-LRRK2, pS129-αSYN, pS63-c-JUN, and CC3. We conclude that the observed neuroprotective effects of PF-475 are due to direct inhibition of LRRK2 activity and that the LRRK2 protein is upstream of the molecular cascade of apoptosis and proteinopathy. Our results suggest that PF-475 is an effective neuroprotective agent against endogenous PSEN1 I416T-induced neurotoxicity in DALNs coexisting with Parkinson's disease markers. Therefore, PF-475 may be of gr","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12074268/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144019546","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Novel Crown Ether-Functionalized Fusidic Acid Butyl Ester: Synthesis, Biological Evaluation, In Silico ADMET, and Molecular Docking Studies.","authors":"Hira Sultan, Nuzhat Arshad, Mehreen Lateef","doi":"10.3390/molecules30092033","DOIUrl":"10.3390/molecules30092033","url":null,"abstract":"<p><p>Crown ethers have gained importance in the field of medicine because of their resemblance to natural ionophores like valinomycin. With the goal of developing new pharmacologically important crown ethers, a novel series of crown ethers linked with Fusidic acid butyl ester <b>10a</b>-<b>d</b> were synthesized and characterized by means of their ¹H NMR, ¹³C NMR DEPT-135, FT-IR, and mass spectrometry. In vitro antioxidant and α-glucosidase inhibition activities of all crown ethers along with the precursor Fusidic acid butyl ester were examined and compared to the standard butylated hydroxyanisole and acarbose, respectively. Compounds (FABE-16-crown-4) <b>10b</b> and (FABE-19-crown-5) <b>10c</b> showed high antioxidant potential with the IC₅₀ = 22.5 ± 0.2 μM and 32.1 ± 0.3 μM, respectively, when compared to the standard BHA (IC₅₀ = 44.2 ± 0.34 μM). To understand the binding mode of the compounds, molecular docking investigations were performed using human antioxidant protein, peroxiredoxin 5. Molecular docking studies revealed higher docking scores (-6.5 and -6.7 kcal/mol) for the highly active compounds <b>10c</b> and <b>10b</b>, respectively, than standard BHA (-5.3 kcal/mol). Synthesized crown ethers exhibited moderate α-glucosidase inhibition with (IC₅₀ = 23.5 ± 0.2 to 76.5 ± 0.1 μM) when compared to acarbose as standard (IC₅₀ = 5.2 ± 0.8 μM). The in silico ADMET predictions indicated that the prepared compounds obeyed (bRO5) and Veber's rule for the acceptance as orally administered drugs and indicated that all the prepared crown ethers exhibited calculated values of drug likeness parameters in acceptable ranges that showed good potential of these molecules for further drug development investigations.</p>","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12073497/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144020209","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Echinacoside Ameliorates UVB-Induced Skin Damage Through Selective Inhibition of the Cutaneous TRPV3 Channel.","authors":"Shilun Mo, Xinying Yue, Yaxuan Qu, Guoji Zhang, Liqin Wang, Xiaoying Sun","doi":"10.3390/molecules30092026","DOIUrl":"10.3390/molecules30092026","url":null,"abstract":"<p><p>Excessive exposure to ultraviolet B (UVB) radiation can lead to skin damage, such as erythema and swelling. Echinacoside is a key effective ingredient of medicinal plant <i>Cistanche deserticola</i> commonly used for therapies and treatments for anti-aging and irradiation-related skin diseases. However, the molecular mechanism underlying the action of echinacoside remains unclear. Here, we report that echinacoside ameliorates UVB-induced skin damage by directly acting on the Ca²⁺-permeable and thermosensitive transient receptor potential vanilloid 3 (TRPV3) channel. Topical application of echinacoside efficaciously suppresses skin lesions induced by UVB radiation in wild-type mice but has no additional benefit in <i>Trpv3</i> knockout mice. In whole-cell patch clamp recordings, echinacoside selectively inhibits TRPV3 channel currents induced by 2-aminoethoxydiphenyl borate in a concentration-dependent manner with an IC₅₀ value of 21.94 ± 1.28 μM. The single-channel patch clamp results show that echinacoside significantly reduces the open probability and open frequency without significantly altering TRPV3 channel unitary conductance. Molecular docking and site-specific mutagenesis indicate that residue T636 on the p-loop and residue T665 on the S6 segment of TRPV3 are critical for echinacoside binding to TRPV3. Taken together, our findings provide a molecular basis for further studies as use of natural echinacoside in irradiation-related skin care therapy, thus establishing a significant role of the TRPV3 channel in acute skin injury.</p>","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12073194/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144002441","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering <i>Escherichia coli</i> Biofilms for Curcumin Production.","authors":"Ana Azevedo, Rita Teixeira-Santos, Luciana C Gomes, Sofia O D Duarte, Gabriel A Monteiro, Filipe J Mergulhão","doi":"10.3390/molecules30092031","DOIUrl":"10.3390/molecules30092031","url":null,"abstract":"<p><p>Biofilms are emerging platforms for the production of valuable compounds. The present study is the first to assess the capacity of <i>Escherichia coli</i> biofilms to produce curcumin through the expression of a biosynthetic pathway involving three genes: 4-coumarate-CoA ligase (<i>4CL</i>), diketide-CoA synthase (<i>DCS</i>), and curcumin synthase (<i>CURS</i>). The effects of chemical induction with isopropyl β-d-1-thiogalactopyranoside (IPTG) and ferulic acid (FA), and the incubation temperature on biofilm formation and curcumin production were evaluated. Biofilms were formed in 12-well microtiter plates over three days and then induced with 1 mM IPTG and FA at 2 or 8 mM. After induction, the samples were incubated for two days at 26 or 30 °C. Total and culturable planktonic and biofilm cells, as well as biofilm thickness and volumetric and specific curcumin production, were assessed on days 3, 4, and 5. The results demonstrated that biofilms produced up to 10-fold higher curcumin levels (0.9-2.2 fg·cell^-1) than their planktonic counterparts (0.1-0.3 fg·cell^-1). The highest specific curcumin production (2.2 fg·cell^-1) was achieved using 8 mM FA. However, no significant differences in curcumin production were observed between the induced samples incubated at the tested temperatures. These results validated the potential of biofilm systems for expressing a complete exogenous biosynthetic pathway using metabolic engineering, particularly for curcumin production.</p>","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12073880/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144014014","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and Characterization of pFluor50, a Fluorogenic-Based Kinetic Assay System for High-Throughput Inhibition Screening and Characterization of Time-Dependent Inhibition and Inhibition Type for Six Human CYPs.","authors":"Pratik Shriwas, Andre Revnew, Sarah Roo, Alex Bender, Kevin Miller, Christopher M Hadad, Thomas R Lane, Sean Ekins, Craig A McElroy","doi":"10.3390/molecules30092032","DOIUrl":"10.3390/molecules30092032","url":null,"abstract":"<p><p>Cytochrome P450s (CYPs) play an integral role in drug and xenobiotic metabolism in humans, and thus, understanding CYP inhibition and/or activation by new therapeutic candidates is an important step in the drug development process. Ideally, CYP inhibition/activation assays should be high-throughput, use commercially available components, allow for analysis of metabolism by the majority of human CYPs, and allow for kinetic analysis of inhibition type and time-dependent inhibition. Here, we developed pFluor50, a 384-well microtiter plate-based fluorogenic kinetic enzyme assay system using substrates metabolized by six human CYPs to generate fluorescent products and determined the Michaelis-Menten kinetics constants (K_M) and product formation rates (V_max) for each substrate-CYP pair. The pFluor50 assay was also used to elucidate inhibition type and time-dependent inhibition for some inhibitors, demonstrating its utility for characterizing the observed inhibition, even mechanism-based inhibition. The pFluor50 assay system developed in this study using commercially available components should be very useful for high-throughput screening and further characterization of potential therapeutic candidates for inhibition/activation with the most prevalent human CYPs.</p>","PeriodicalId":19041,"journal":{"name":"Molecules","volume":"30 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12074421/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144036041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}