Pub Date : 2024-08-28DOI: 10.1101/2024.08.28.610072
Colleen M Pike, James A Levi, Lauren A Boone, Swetha Peddibhotla, Jacob Johnson, Bailey Zwarycz, Maureen K Bunger, William Thelin, Elizabeth M Boazak
Gastrointestinal toxicities (GITs) are the most prevalent adverse events (AE) reported in clinical trials, often resulting in dose-limitations that reduce drug efficacy and delay development and treatment optimization. Preclinical animal models do not accurately replicate human GI physiology, leaving few options for early detection of GI side effects prior to human studies. Development of an accurate model that predicts GIT earlier in drug discovery programs would better support successful clinical trial outcomes. Chemotherapeutics, which exhibit high rates of clinical GIT, frequently target mitotic cells. Therefore, we hypothesized that a model utilizing proliferative cell populations derived from human intestinal crypts would predict the occurrence of clinical GITs with high accuracy. Here, we describe the development of a multiparametric assay utilizing the RepliGut Planar system, an intestinal stem cell-derived platform cultured in an accessible high throughput Transwell format. This assay addresses key physiological elements of GIT by assessing cell proliferation (EdU incorporation), cell abundance (DAPI quantification), and barrier function (TEER). Using this approach, we demonstrate that primary proliferative cell populations reproducibly respond to marketed chemotherapeutics at physiologic concentrations. To determine the ability of this model to predict clinical diarrhea risk, we evaluated a set of 30 drugs with known clinical diarrhea incidence in three human donors, comparing results to known plasma drug concentrations. This resulted in highly accurate predictions of diarrhea potential for each endpoint (balanced accuracy of 91% for DAPI, 90% for EdU, 88% for TEER) with minimal variation across human donors. In vitro toxicity screening using primary proliferative cells may enable improved safety evaluations, reducing the risk of AEs in clinical trials and ultimately lead to safer and more effective treatments for patients.
{"title":"High-Throughput Assay for Predicting Diarrhea Risk Using a 2D Human Intestinal Stem Cell-Derived Model","authors":"Colleen M Pike, James A Levi, Lauren A Boone, Swetha Peddibhotla, Jacob Johnson, Bailey Zwarycz, Maureen K Bunger, William Thelin, Elizabeth M Boazak","doi":"10.1101/2024.08.28.610072","DOIUrl":"https://doi.org/10.1101/2024.08.28.610072","url":null,"abstract":"Gastrointestinal toxicities (GITs) are the most prevalent adverse events (AE) reported in clinical trials, often resulting in dose-limitations that reduce drug efficacy and delay development and treatment optimization. Preclinical animal models do not accurately replicate human GI physiology, leaving few options for early detection of GI side effects prior to human studies. Development of an accurate model that predicts GIT earlier in drug discovery programs would better support successful clinical trial outcomes. Chemotherapeutics, which exhibit high rates of clinical GIT, frequently target mitotic cells. Therefore, we hypothesized that a model utilizing proliferative cell populations derived from human intestinal crypts would predict the occurrence of clinical GITs with high accuracy. Here, we describe the development of a multiparametric assay utilizing the RepliGut Planar system, an intestinal stem cell-derived platform cultured in an accessible high throughput Transwell format. This assay addresses key physiological elements of GIT by assessing cell proliferation (EdU incorporation), cell abundance (DAPI quantification), and barrier function (TEER). Using this approach, we demonstrate that primary proliferative cell populations reproducibly respond to marketed chemotherapeutics at physiologic concentrations. To determine the ability of this model to predict clinical diarrhea risk, we evaluated a set of 30 drugs with known clinical diarrhea incidence in three human donors, comparing results to known plasma drug concentrations. This resulted in highly accurate predictions of diarrhea potential for each endpoint (balanced accuracy of 91% for DAPI, 90% for EdU, 88% for TEER) with minimal variation across human donors. In vitro toxicity screening using primary proliferative cells may enable improved safety evaluations, reducing the risk of AEs in clinical trials and ultimately lead to safer and more effective treatments for patients.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"3 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-28DOI: 10.1101/2024.08.27.610017
Jivianne Lee, Saroj K Basak, Hong-Ho Yang, Kimberly Sullivan, Tom Maxim, Daniel S Shin, Nancy Klimas, Eri Srivatsan
Background: N,N-Diethyl-Meta-Toluamide (DEET) and permethrin are pesticides commonly used in combination due to their synergistic insecticidal and repellent properties. This study investigates whether simultaneous exposure to these compounds elicits synergistic cytotoxicity in sinonasal epithelial cells (SNECs). Material and Methods: Ethmoid sinus mucosal specimens were procured from eight patients during endoscopic sinus surgery. SNECs were expanded on culture plates and exposed to various concentrations of DEET and permethrin (0-5μm), individually and concurrently, for up to 156 hours. Experiments were replicated in triplicates and cell viability was recorded every 2 hours using IncuCyte real-time cell imaging system. Synergy score was calculated on the basis of Loewe additivity synergy finder model. Results: DEET and permethrin exhibited synergistic cytotoxicity across all eight tissues, albeit with variations in onset and magnitude. Peak synergy was observed at 144h for tissue 1 (SLoewe 4.2, 95% CI 1.8-7.3; [permethrin concentration, DEET concentration] [2.5μM, 1.25μM]), 48h for tissue 2 (19.9, 16.5-23.6; [1.25, 0.625]), 144h for tissue 3 (15.2, 3.9-30.4; [0.625, 1.25]), 144h for tissue 4 (6.4, -3.6 to 18.0; [0.625, 0.625]), 48h for tissue 5 (10.1, 8.9-12.5; [0.625, 1.25]), 96h for tissue 6 (24.7, 12.2-36.3; [0.625, 0.625]), 48h for tissue 7 (47.7, 29.6-62.2; [0.625, 1.25]), and 96h for 8 (47.4, 26.8-67.0; [0.625, 0.625]). Conclusion: The concurrent exposure of DEET and permethrin can lead to synergistic cytotoxicity in sinonasal epithelia. Further research is warranted in preclinical animal models to explore whether this synergy accelerates the pathogenesis of chronic rhinosinusitis.
{"title":"Synergistic Cytotoxicity of Permethrin and N,N-Diethyl-Meta-Toluamide on Sinonasal Epithelial Cells","authors":"Jivianne Lee, Saroj K Basak, Hong-Ho Yang, Kimberly Sullivan, Tom Maxim, Daniel S Shin, Nancy Klimas, Eri Srivatsan","doi":"10.1101/2024.08.27.610017","DOIUrl":"https://doi.org/10.1101/2024.08.27.610017","url":null,"abstract":"Background: N,N-Diethyl-Meta-Toluamide (DEET) and permethrin are pesticides commonly used in combination due to their synergistic insecticidal and repellent properties. This study investigates whether simultaneous exposure to these compounds elicits synergistic cytotoxicity in sinonasal epithelial cells (SNECs). Material and Methods: Ethmoid sinus mucosal specimens were procured from eight patients during endoscopic sinus surgery. SNECs were expanded on culture plates and exposed to various concentrations of DEET and permethrin (0-5μm), individually and concurrently, for up to 156 hours. Experiments were replicated in triplicates and cell viability was recorded every 2 hours using IncuCyte real-time cell imaging system. Synergy score was calculated on the basis of Loewe additivity synergy finder model. Results: DEET and permethrin exhibited synergistic cytotoxicity across all eight tissues, albeit with variations in onset and magnitude. Peak synergy was observed at 144h for tissue 1 (SLoewe 4.2, 95% CI 1.8-7.3; [permethrin concentration, DEET concentration] [2.5μM, 1.25μM]), 48h for tissue 2 (19.9, 16.5-23.6; [1.25, 0.625]), 144h for tissue 3 (15.2, 3.9-30.4; [0.625, 1.25]), 144h for tissue 4 (6.4, -3.6 to 18.0; [0.625, 0.625]), 48h for tissue 5 (10.1, 8.9-12.5; [0.625, 1.25]), 96h for tissue 6 (24.7, 12.2-36.3; [0.625, 0.625]), 48h for tissue 7 (47.7, 29.6-62.2; [0.625, 1.25]), and 96h for 8 (47.4, 26.8-67.0; [0.625, 0.625]). Conclusion: The concurrent exposure of DEET and permethrin can lead to synergistic cytotoxicity in sinonasal epithelia. Further research is warranted in preclinical animal models to explore whether this synergy accelerates the pathogenesis of chronic rhinosinusitis.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-28DOI: 10.1101/2024.08.28.610063
Katariina Seppala, Ines Reigada, Olli Matilainen, Tomi Rantamaki, Leena Hanski
Transparency of C. elegans enables microscopic in vivo imaging of cellular processes, but immobilization is required due to high locomotor activity. Preservative NaN3 is commonly used for this, but is associated with oxidative stress and toxicity. Here, anesthetic-like effects of dissociate anesthetic ketamine in C. elegans are presented using video recordings and infrared-based automated activity tracking. Ketamine caused a reversible blockade of locomotion at a similar concentration (20-50 mM) at which NaN3 produces paralysis. Moreover, the recovery rate was remarkably high, and short-term ketamine treatment did not show signs of SKN-1 activation, a marker of the stress response.
{"title":"Anesthetic-like effects of ketamine in C. elegans","authors":"Katariina Seppala, Ines Reigada, Olli Matilainen, Tomi Rantamaki, Leena Hanski","doi":"10.1101/2024.08.28.610063","DOIUrl":"https://doi.org/10.1101/2024.08.28.610063","url":null,"abstract":"Transparency of C. elegans enables microscopic in vivo imaging of cellular processes, but immobilization is required due to high locomotor activity. Preservative NaN3 is commonly used for this, but is associated with oxidative stress and toxicity. Here, anesthetic-like effects of dissociate anesthetic ketamine in C. elegans are presented using video recordings and infrared-based automated activity tracking. Ketamine caused a reversible blockade of locomotion at a similar concentration (20-50 mM) at which NaN3 produces paralysis. Moreover, the recovery rate was remarkably high, and short-term ketamine treatment did not show signs of SKN-1 activation, a marker of the stress response.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"24 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184264","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-28DOI: 10.1101/2024.08.27.609990
Jacqueline A. Barnett, Jessica K Josephson, Natasha A. Haskey, Miranda M Hart, Kiran K. Soma, Maya L. Bandy, Carson J. McComb, Andrea Verdugo, Sanjoy Ghosh, Clara Letef, Amanda Copp, Julien Gibon, Jiayu Ye, Ryland T. Giebelhaus, Susan J. Murch, Minseon M. Jung, Deanna L. Gibson
Glyphosate, a widely used herbicide in North America, has become prevalent in the food supply due to preharvest applications, raising concerns about potential health impacts. This study investigated the effects of prenatal glyphosate exposure on the gut bacteriome, colitis, metabolic health, and behavior across generations in mice. In healthy mice, glyphosate decreased goblet cell numbers and mucin-2 expression in the colon and dysregulated cytokines in F1 and F2 progeny at levels below the EPA acceptable daily limit. Glyphosate also disrupted metabolism, including impaired glucose tolerance, increased insulin resistance and reduced serum GLP-1 levels. Moreover, prenatal glyphosate exposure induced behavioral deficits, including reduced locomotor activity and impaired working memory, which are associated with alterations in the gut microbiome composition and key gut-brain axis mediators. These data underscore the potential risks associated with glyphosate exposure, highlighting the need for further research and regulatory consideration.
{"title":"Prenatal glyphosate exposure disrupts the gut-brain axis across several generations in mice.","authors":"Jacqueline A. Barnett, Jessica K Josephson, Natasha A. Haskey, Miranda M Hart, Kiran K. Soma, Maya L. Bandy, Carson J. McComb, Andrea Verdugo, Sanjoy Ghosh, Clara Letef, Amanda Copp, Julien Gibon, Jiayu Ye, Ryland T. Giebelhaus, Susan J. Murch, Minseon M. Jung, Deanna L. Gibson","doi":"10.1101/2024.08.27.609990","DOIUrl":"https://doi.org/10.1101/2024.08.27.609990","url":null,"abstract":"Glyphosate, a widely used herbicide in North America, has become prevalent in the food supply due to preharvest applications, raising concerns about potential health impacts. This study investigated the effects of prenatal glyphosate exposure on the gut bacteriome, colitis, metabolic health, and behavior across generations in mice. In healthy mice, glyphosate decreased goblet cell numbers and mucin-2 expression in the colon and dysregulated cytokines in F1 and F2 progeny at levels below the EPA acceptable daily limit. Glyphosate also disrupted metabolism, including impaired glucose tolerance, increased insulin resistance and reduced serum GLP-1 levels. Moreover, prenatal glyphosate exposure induced behavioral deficits, including reduced locomotor activity and impaired working memory, which are associated with alterations in the gut microbiome composition and key gut-brain axis mediators. These data underscore the potential risks associated with glyphosate exposure, highlighting the need for further research and regulatory consideration.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"10 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184261","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-27DOI: 10.1101/2024.08.27.607356
Silas L Wurnig, Max E Huber, Corinna Weiler, Hanna Baltrukevich, Nicole Merten, Isabel Stoetzel, Yinshui Chang, Rene Klammer, Dirk Baumjohann, Eva Kiermaier, Peter Kolb, Evi Kostenis, Matthias Schiedel, Finn K Hansen
Intracellularly acting ligands of G protein-coupled receptors (GPCRs) are gaining significant interest in GPCR drug discovery. In this study, we report the development of the fluorescent ligand Mz437 (4) targeting the CC chemokine receptor CCR7 at an intracellular allosteric site. We demonstrate its experimental power by applying 4 to identify two improved intracellular CCR7 antagonists, SLW131 (10) and SLW132 (21m), developed by converting two weakly active antagonists into single- or double-digit nanomolar ligands with minimal modifications. The thiadiazoledioxide 10 was derived from the CCR7 antagonist Cmp2105 by removing a methyl group from the benzamide moiety, while the squaramide 21m was obtained from the CXCR1/CXCR2 antagonist and clinical candidate navarixin by replacing the ethyl substituent by a tert-butyl group to engage a lipophilic subpocket. We show that 10 and 21m qualify to probe CCR7 biology both in recombinant cells and in the endogenous signaling environment of immune cells. Our novel probes are expected to facilitate the design of next-generation intracellular CCR7 ligands and serve as molecular tools to interrogate CCR7 biology in human and murine endogenous settings.
{"title":"A fluorescent probe enables the discovery of improved antagonists targeting the intracellular allosteric site of the chemokine receptor CCR7","authors":"Silas L Wurnig, Max E Huber, Corinna Weiler, Hanna Baltrukevich, Nicole Merten, Isabel Stoetzel, Yinshui Chang, Rene Klammer, Dirk Baumjohann, Eva Kiermaier, Peter Kolb, Evi Kostenis, Matthias Schiedel, Finn K Hansen","doi":"10.1101/2024.08.27.607356","DOIUrl":"https://doi.org/10.1101/2024.08.27.607356","url":null,"abstract":"Intracellularly acting ligands of G protein-coupled receptors (GPCRs) are gaining significant interest in GPCR drug discovery. In this study, we report the development of the fluorescent ligand Mz437 (4) targeting the CC chemokine receptor CCR7 at an intracellular allosteric site. We demonstrate its experimental power by applying 4 to identify two improved intracellular CCR7 antagonists, SLW131 (10) and SLW132 (21m), developed by converting two weakly active antagonists into single- or double-digit nanomolar ligands with minimal modifications. The thiadiazoledioxide 10 was derived from the CCR7 antagonist Cmp2105 by removing a methyl group from the benzamide moiety, while the squaramide 21m was obtained from the CXCR1/CXCR2 antagonist and clinical candidate navarixin by replacing the ethyl substituent by a tert-butyl group to engage a lipophilic subpocket. We show that 10 and 21m qualify to probe CCR7 biology both in recombinant cells and in the endogenous signaling environment of immune cells. Our novel probes are expected to facilitate the design of next-generation intracellular CCR7 ligands and serve as molecular tools to interrogate CCR7 biology in human and murine endogenous settings.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"67 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184263","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-26DOI: 10.1101/2024.08.25.609588
Ryuto Tamura, Yusuke Ujihara, Mizuki Yamamoto, Shuntaro Matsushima, Haruki Kasahara, Daiki Nasukawa, Kazuko Hayashi, Kota Yamada, Masashi Tanaka, Koji Toda
Vocal communication is essential for survival in many animals, including humans. Mice produce ultrasonic vocalizations (USVs) in various social contexts. N-methyl-d-aspartate (NMDA) receptors are widely distributed in the central nervous system and play essential roles in synaptic plasticity, learning, and behavior. However, the role of NMDA receptors in USVs remains unclear. Here, we examined the effects of systemic administration of a noncompetitive NMDA antagonist, MK-801, on USVs during courtship and in other social and nonsocial behaviors. First, we investigated the USVs emitted by male mice. After the successful classification of USVs into 11 categories using VocalMat, an open-source computational vision and machine-learning technique, we analyzed the relationship between USV properties and approaching behavior toward female mice. Male mice produced USVs when they were close to female mice, which confirmed that USVs were associated with social signals. Second, we examined the effects of intraperitoneal injection of MK-801 on USVs and approach behavior. Intraperitoneal injection of MK-801 dose-dependently decreased the number of USVs and increased locomotor activity, but did not affect the time spent with female mice. In additional experiments, we confirmed that the injection of MK-801 decreased the time spent with a novel same-sex individual in the social preference task, increased locomotor activity, and inhibited excretion in the open-field task. Taken together, these results demonstrate that NMDA receptors play important roles in vocal communication and social behavior.
{"title":"Disruption of ultrasonic vocalization with systemic administration of the non-competitive N-methyl-d-aspartate receptor antagonist MK-801 in adult male mice","authors":"Ryuto Tamura, Yusuke Ujihara, Mizuki Yamamoto, Shuntaro Matsushima, Haruki Kasahara, Daiki Nasukawa, Kazuko Hayashi, Kota Yamada, Masashi Tanaka, Koji Toda","doi":"10.1101/2024.08.25.609588","DOIUrl":"https://doi.org/10.1101/2024.08.25.609588","url":null,"abstract":"Vocal communication is essential for survival in many animals, including humans. Mice produce ultrasonic vocalizations (USVs) in various social contexts. N-methyl-d-aspartate (NMDA) receptors are widely distributed in the central nervous system and play essential roles in synaptic plasticity, learning, and behavior. However, the role of NMDA receptors in USVs remains unclear. Here, we examined the effects of systemic administration of a noncompetitive NMDA antagonist, MK-801, on USVs during courtship and in other social and nonsocial behaviors. First, we investigated the USVs emitted by male mice. After the successful classification of USVs into 11 categories using VocalMat, an open-source computational vision and machine-learning technique, we analyzed the relationship between USV properties and approaching behavior toward female mice. Male mice produced USVs when they were close to female mice, which confirmed that USVs were associated with social signals. Second, we examined the effects of intraperitoneal injection of MK-801 on USVs and approach behavior. Intraperitoneal injection of MK-801 dose-dependently decreased the number of USVs and increased locomotor activity, but did not affect the time spent with female mice. In additional experiments, we confirmed that the injection of MK-801 decreased the time spent with a novel same-sex individual in the social preference task, increased locomotor activity, and inhibited excretion in the open-field task. Taken together, these results demonstrate that NMDA receptors play important roles in vocal communication and social behavior.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"26 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-25DOI: 10.1101/2024.08.23.609337
Maja Stevanoska, Karsten Beekmann, Ans Punt, Shana Sturla, Georg Aichinger
Hop extracts containing prenylated polyphenols such as 8-prenylnaringenin (8-PN) and its precursor isoxanthohumol (iXN) are popular among women seeking natural alternatives to hormone therapy for postmenopausal symptoms. Due to structural similarities with estrogens, these compounds act as estrogen receptor agonists. Especially 8-PN, described as the most potent phytoestrogen known to date, poses a potential risk for endocrine disruption. Therefore, its use as a hormone replacement raises concerns for human health. However, a significant challenge in assessing the potential endocrine-disruptive effects of hop polyphenols is the lack of data on their toxicokinetics. Particularly, information on in vivo concentrations in target tissues is lacking. To address this gap, we developed a physiologically based kinetic (PBK) model tailored to female physiology. The model was used to predict the levels of hop polyphenols in human blood and target tissues under realistic exposure scenarios. The predictions suggest that iXN and 8-PN concentrations in target tissues reach the low nanomolar range after dietary supplementation. This study enhances our understanding of the safety profile of hop polyphenols and highlights the need for further research into their use as an alternative to hormone therapy in menopausal women.
{"title":"Predicting in vivo concentrations of dietary hop phytoestrogens by physiologically based kinetic modeling","authors":"Maja Stevanoska, Karsten Beekmann, Ans Punt, Shana Sturla, Georg Aichinger","doi":"10.1101/2024.08.23.609337","DOIUrl":"https://doi.org/10.1101/2024.08.23.609337","url":null,"abstract":"Hop extracts containing prenylated polyphenols such as 8-prenylnaringenin (8-PN) and its precursor isoxanthohumol (iXN) are popular among women seeking natural alternatives to hormone therapy for postmenopausal symptoms. Due to structural similarities with estrogens, these compounds act as estrogen receptor agonists. Especially 8-PN, described as the most potent phytoestrogen known to date, poses a potential risk for endocrine disruption. Therefore, its use as a hormone replacement raises concerns for human health. However, a significant challenge in assessing the potential endocrine-disruptive effects of hop polyphenols is the lack of data on their toxicokinetics. Particularly, information on in vivo concentrations in target tissues is lacking. To address this gap, we developed a physiologically based kinetic (PBK) model tailored to female physiology. The model was used to predict the levels of hop polyphenols in human blood and target tissues under realistic exposure scenarios. The predictions suggest that iXN and 8-PN concentrations in target tissues reach the low nanomolar range after dietary supplementation. This study enhances our understanding of the safety profile of hop polyphenols and highlights the need for further research into their use as an alternative to hormone therapy in menopausal women.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"3 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142227645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-23DOI: 10.1101/2024.08.20.608791
Urban Fagerholm, Sven Hellberg, Jonathan Alvarsson, Morgan Ekmefjord, Ola Spjuth
Background - The influential Lipinskis Rule of 5 (Ro5) describes molecular properties important for oral absorption in humans. According to Ro5, poor absorption is more likely when 2 or more of its criteria (molecular weight (MW) above 500 g/mol, calculated octanol-water partition coefficient (clog P) above 5, above 5 hydrogen bond donors (HBD) and above 10 hydrogen bond acceptors (HBA)) are violated. Earlier evaluations have shown that many drugs are sufficiently well absorbed into the systemic circulation despite many Ro5-violations. No evaluation of Ro5 vs fraction absorbed (fa) has, however, been done. Methods - Datasets of orally administered drugs violating and not violating Ro5 and with available human clinical fa-values were assembled, and contrasted to machine learning based predictions using the ANDROMEDA prediction software having a major MW-domain of 150-750 g/mol. Results - 129 Ro5-violent compounds (29 with MW above 1000 g/mol) were found, 59 of which had fa-values (42 % mean fa). 34 % and 66 % of compounds were predicted as having fa below 10 % and above 10-30 % respectively, which was in good agreement with measured fa of 37 % and 63 %. The fa for all compounds with fa above 5 % and above 10 % were correctly predicted. For compounds with fa above 30 %, 81 % were predicted to have a fa above 30 %, but none were predicted to have a fa below 10 %. The Q2 for predicted vs observed fa was 0.64. For a set of 77 compounds without Ro5 violation (80 % mean fa), all compounds were correctly predicted to have a fa below or above 30 % (Q2=0.56). Among these are compounds with poor uptake (below 1 % to 7 %). Conclusion - We show that machine learning based predictions of fa are superior to Ro5 for assessing oral absorption obstacles in humans. Too strict reliance on Ro5 may hence constitute a risk. ANDROMEDA predicts fa well, easily and quickly, and also differentiates well between poor and adequate oral uptake for compounds violating and not-violating Ro5. This makes it a valid and useful tool capable of predicting oral absorption in humans with good accuracy and replacing Ro5 for oral absorption assessments.
背景--颇具影响力的利平斯基5法则（Ro5）描述了对人体口服吸收非常重要的分子特性。根据 Ro5，如果违反了其中的两个或两个以上标准（分子量（MW）大于 500 g/mol、计算的辛醇-水分配系数（clog P）大于 5、氢键供体（HBD）大于 5 和氢键受体（HBA）大于 10），则药物更有可能吸收不良。先前的评估表明，尽管有许多药物违反了 Ro5，但它们仍能被全身循环充分吸收。但是，还没有对 Ro5 与吸收率 (fa) 进行过评估。方法 - 收集了违反和未违反 Ro5 规定的口服药物数据集，并提供了人体临床 fa 值，将其与使用 ANDROMEDA 预测软件进行的基于机器学习的预测结果进行对比，该软件的主要截面积为 150-750 g/mol。根据预测，34% 和 66% 的化合物的 fa 值分别低于 10% 和高于 10-30%，这与 37% 和 63% 的实测 fa 值十分吻合。所有 fa 高于 5 % 和高于 10 % 的化合物的 fa 都被正确预测。对于fa高于30%的化合物，81%的化合物被预测为fa高于30%，但没有化合物被预测为fa低于10%。预测 fa 与观察 fa 的 Q2 值为 0.64。对于一组没有违反 Ro5 的 77 种化合物（平均 fa 为 80%），所有化合物都被正确预测为 fa 低于或高于 30%（Q2=0.56）。结论 - 我们的研究表明，在评估人体口服吸收障碍方面，基于机器学习的 fa 预测优于 Ro5。因此，过于严格地依赖 Ro5 可能会带来风险。ANDROMEDA 可以很好地预测 fa，简单快捷，还能很好地区分违反和未违反 Ro5 的化合物口服吸收不良和充分。这使其成为一种有效、实用的工具，能够准确预测人体的口服吸收，并取代 Ro5 进行口服吸收评估。
{"title":"Comparing Lipinskis Rule of 5 and Machine Learning Based Prediction of Fraction Absorbed for Assessing Oral Absorption in Humans","authors":"Urban Fagerholm, Sven Hellberg, Jonathan Alvarsson, Morgan Ekmefjord, Ola Spjuth","doi":"10.1101/2024.08.20.608791","DOIUrl":"https://doi.org/10.1101/2024.08.20.608791","url":null,"abstract":"Background - The influential Lipinskis Rule of 5 (Ro5) describes molecular properties important for oral absorption in humans. According to Ro5, poor absorption is more likely when 2 or more of its criteria (molecular weight (MW) above 500 g/mol, calculated octanol-water partition coefficient (clog P) above 5, above 5 hydrogen bond donors (HBD) and above 10 hydrogen bond acceptors (HBA)) are violated. Earlier evaluations have shown that many drugs are sufficiently well absorbed into the systemic circulation despite many Ro5-violations. No evaluation of Ro5 vs fraction absorbed (fa) has, however, been done. Methods - Datasets of orally administered drugs violating and not violating Ro5 and with available human clinical fa-values were assembled, and contrasted to machine learning based predictions using the ANDROMEDA prediction software having a major MW-domain of 150-750 g/mol.\u0000Results - 129 Ro5-violent compounds (29 with MW above 1000 g/mol) were found, 59 of which had fa-values (42 % mean fa). 34 % and 66 % of compounds were predicted as having fa below 10 % and above 10-30 % respectively, which was in good agreement with measured fa of 37 % and 63 %. The fa for all compounds with fa above 5 % and above 10 % were correctly predicted. For compounds with fa above 30 %, 81 % were predicted to have a fa above 30 %, but none were predicted to have a fa below 10 %. The Q2 for predicted vs observed fa was 0.64. For a set of 77 compounds without Ro5 violation (80 % mean fa), all compounds were correctly predicted to have a fa below or above 30 % (Q2=0.56). Among these are compounds with poor uptake (below 1 % to 7 %).\u0000Conclusion - We show that machine learning based predictions of fa are superior to Ro5 for assessing oral absorption obstacles in humans. Too strict reliance on Ro5 may hence constitute a risk. ANDROMEDA predicts fa well, easily and quickly, and also differentiates well between poor and adequate oral uptake for compounds violating and not-violating Ro5. This makes it a valid and useful tool capable of predicting oral absorption in humans with good accuracy and replacing Ro5 for oral absorption assessments.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"30 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Back ground: Litsea glutinosa leaves (LG) have been used as traditional medicine to treat diseases since ancient times. This study aimed to investigate the cold methanol extract of LG for its analgesic, antipyretic and antibacterial activities, along with preliminary phytochemical screening and acute toxicity test. Methods: In this study, we first investigated the major phytochemicals group present applying modern chromatographic technique HPLC in addition to conventional methods for Phyto screening in cold methanol extracts LG leaves. Both methods simultaneously demonstrated major bioactive compounds found in the extract were phenol, flavonoid. The purposive efficacy and toxicity were then assessed through preclinical testing. Results: In hot plate method, the highest pain inhibitory activity was found at a dose of 500 mg/kg of crude extract (3.37± 0.31sec) which differed significantly (P <0.01 and P <0.001) with that of the standard drug morphine (6.47± 0.23 sec). The extract significantly prolonged reaction latency to thermal-induced pain in hotplate model. Analgesic activity at 500 mg/kg, LG extract produced a 70% suppression of writhing in mice, which was statistically significant (p<0.001) compared to standard morphine's (77.5%) inhibition. In antipyretic activity assay, the crude extract showed notable reduction in body temperature (36.17 ± 0.32°C) at dose of 300 mg/kg-body weight, when the standard (at dose 100 mg/kg-body weight) exerted (36.32 ± 0.67°C) after 3 h of administration. In antibacterial studies, results showed that inhibition of bacterial growth at 400 μg dose of each extract clearly inhibited growth of bacteria from 11 to 22 mm. The extractives carbon tetrachloride fraction, chloroform soluble fraction, ethyl acetate fraction demonstrated notably greater inhibitory zone widths (p < 0.05) against tested strains. Conclusion: Overall, the cold methanol extract of LG leaves demonstrates the therapeutic potential in preclinical settings.
{"title":"Phytochemical screening, analgesic, anti-pyretic and antibacterial potentials of Litsea glutinosa (L) leaves extracts in vivo and in vitro technique","authors":"Zubair Labu, Samira Karim, Md. Tarekur Rahman, Md. Imran Hossain, Md Shakil","doi":"10.1101/2024.08.21.609048","DOIUrl":"https://doi.org/10.1101/2024.08.21.609048","url":null,"abstract":"Back ground: Litsea glutinosa leaves (LG) have been used as traditional medicine to treat diseases since ancient times. This study aimed to investigate the cold methanol extract of LG for its analgesic, antipyretic and antibacterial activities, along with preliminary phytochemical screening and acute toxicity test. Methods: In this study, we first investigated the major phytochemicals group present applying modern chromatographic technique HPLC in addition to conventional methods for Phyto screening in cold methanol extracts LG leaves. Both methods simultaneously demonstrated major bioactive compounds found in the extract were phenol, flavonoid. The purposive efficacy and toxicity were then assessed through preclinical testing. Results: In hot plate method, the highest pain inhibitory activity was found at a dose of 500 mg/kg of crude extract (3.37± 0.31sec) which differed significantly (P <0.01 and P <0.001) with that of the standard drug morphine (6.47± 0.23 sec). The extract significantly prolonged reaction latency to thermal-induced pain in hotplate model. Analgesic activity at 500 mg/kg, LG extract produced a 70% suppression of writhing in mice, which was statistically significant (p<0.001) compared to standard morphine's (77.5%) inhibition. In antipyretic activity assay, the crude extract showed notable reduction in body temperature (36.17 ± 0.32°C) at dose of 300 mg/kg-body weight, when the standard (at dose 100 mg/kg-body weight) exerted (36.32 ± 0.67°C) after 3 h of administration. In antibacterial studies, results showed that inhibition of bacterial growth at 400 μg dose of each extract clearly inhibited growth of bacteria from 11 to 22 mm. The extractives carbon tetrachloride fraction, chloroform soluble fraction, ethyl acetate fraction demonstrated notably greater inhibitory zone widths (p < 0.05) against tested strains. Conclusion: Overall, the cold methanol extract of LG leaves demonstrates the therapeutic potential in preclinical settings.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"35 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184295","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Statins, HMG-CoA reductase inhibitors, have been widely prescribed to lower circulating low-density lipoprotein cholesterol levels and reduce the risk of cardiovascular disease. Although statins are well tolerated, statin-associated muscle symptoms (SAMS) are the major adverse effect and cause statin intolerance. Therefore, understanding the molecular mechanisms of SAMS and identifying effective strategies for its prevention are of significant clinical importance; however, both remain unclear. Here we establish a model of statin-induced myopathy (SIM) with human induced pluripotent stem cell (hiPSC)-derived myocytes (iPSC-MCs) and investigate the effect of statins on protein homeostasis (proteostasis) that affects skeletal muscle wasting and myotoxicity. We show that treating hiPSC-MCs with statins induces atrophic phenotype and myotoxicity, establishing a hiPSC-based SIM model. We then examine whether statins impair the balance between protein synthesis and degradation. The results show that statins not only suppress protein synthesis but also promote protein degradation by upregulating the expression of the muscle-specific E3 ubiquitin ligase Atrogin-1 in a mevalonate pathway-dependent manner. Mechanistically, blocking the mevalonate pathway inactivates the protein kinase Akt, leading to the inhibition of mTORC1 and GSK3β but the activation of FOXO1. These changes explain the statin-induced impairment in proteostasis. Finally, we show that pharmacological blockage of FOXO1 prevents SIM in hiPSC-MCs, implicating FOXO1 as a key mediator of SIM. Taken together, this study suggests that the mevalonate pathway is critical for maintaining skeletal muscle proteostasis and identifies FOXO1 as a potential target for preventing SIM.
{"title":"Modeling statin-induced myopathy with human iPSCs reveals that impaired proteostasis underlies the myotoxicity and is targetable for the prevention","authors":"Xiaolin Zhao, Liyang Ni, Miharu Kubo, Mariko Matsuto, Hidetoshi Sakurai, Makoto Shimizu, Yu Takahashi, Ryuichiro Sato, Yoshio Yamauchi","doi":"10.1101/2024.08.23.608904","DOIUrl":"https://doi.org/10.1101/2024.08.23.608904","url":null,"abstract":"Statins, HMG-CoA reductase inhibitors, have been widely prescribed to lower circulating low-density lipoprotein cholesterol levels and reduce the risk of cardiovascular disease. Although statins are well tolerated, statin-associated muscle symptoms (SAMS) are the major adverse effect and cause statin intolerance. Therefore, understanding the molecular mechanisms of SAMS and identifying effective strategies for its prevention are of significant clinical importance; however, both remain unclear. Here we establish a model of statin-induced myopathy (SIM) with human induced pluripotent stem cell (hiPSC)-derived myocytes (iPSC-MCs) and investigate the effect of statins on protein homeostasis (proteostasis) that affects skeletal muscle wasting and myotoxicity. We show that treating hiPSC-MCs with statins induces atrophic phenotype and myotoxicity, establishing a hiPSC-based SIM model. We then examine whether statins impair the balance between protein synthesis and degradation. The results show that statins not only suppress protein synthesis but also promote protein degradation by upregulating the expression of the muscle-specific E3 ubiquitin ligase Atrogin-1 in a mevalonate pathway-dependent manner. Mechanistically, blocking the mevalonate pathway inactivates the protein kinase Akt, leading to the inhibition of mTORC1 and GSK3β but the activation of FOXO1. These changes explain the statin-induced impairment in proteostasis. Finally, we show that pharmacological blockage of FOXO1 prevents SIM in hiPSC-MCs, implicating FOXO1 as a key mediator of SIM. Taken together, this study suggests that the mevalonate pathway is critical for maintaining skeletal muscle proteostasis and identifies FOXO1 as a potential target for preventing SIM.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"142 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184275","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：