Discovery of rhynchophylline and mitraphylline in two Thai Mitragyna species and the investigation of their biological activity via opioid gene expression analysis

Abstract Mitragyna speciosa (Ms), M. diversifolia (Md), M. hirsuta (Mh) and M. rotundifolia (Mr) were investigated for phytochemicals by GC-MS and GC-FID, cytotoxicity and genotoxicity testing by MTT and comet assay, and biological activity examination through gene expression of human µ, δ, κ, and n...

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Main Authors: Runglawan Sudmoon, Tawatchai Tanee, Warin Wonok, Unchaleeporn Ameamsri, Thomas Liehr, Sakda Daduang, Pornnarong Siripiyasing, Arunrat Chaveerach
Format: Article
Language:English
Published: Nature Portfolio 2025-02-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-89715-5
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author Runglawan Sudmoon
Tawatchai Tanee
Warin Wonok
Unchaleeporn Ameamsri
Thomas Liehr
Sakda Daduang
Pornnarong Siripiyasing
Arunrat Chaveerach
author_facet Runglawan Sudmoon
Tawatchai Tanee
Warin Wonok
Unchaleeporn Ameamsri
Thomas Liehr
Sakda Daduang
Pornnarong Siripiyasing
Arunrat Chaveerach
author_sort Runglawan Sudmoon
collection DOAJ
description Abstract Mitragyna speciosa (Ms), M. diversifolia (Md), M. hirsuta (Mh) and M. rotundifolia (Mr) were investigated for phytochemicals by GC-MS and GC-FID, cytotoxicity and genotoxicity testing by MTT and comet assay, and biological activity examination through gene expression of human µ, δ, κ, and nociceptin opioid receptors by qRT-PCR. The opioid substances mitragynine, 7-hydroxymitragynine, and mitraphylline were found in all studied species, and, for first time, rhynchophylline was found in Mr, and mitraphylline in Md, Mh and Mr. The MTT and comet assays of the ethanol and hexane leaf extracts on PBMCs revealed no cytotoxicity and no significant genotoxicity compared to the negative control, except for the hexane leaf extract of Mh, which caused significant DNA damage. The biological activity of the ethanolic extract of the four species showed a binding affinity to the µ (MOR) receptor revealing a relative gene expression of 89.54 and 50.41 by Ms and Md at 1.92 and 1.133 mg/ml, 32.42 and 19.97 mg/ml by Md and Ms at 3.77 and 1.76 mg/ml. Mr contained the three opioids mentioned plus rhynchophylline and showed low relative µ (MOR) gene expression of 16.89 at 0.189 mg/ml, while as an additional species, Ipomoea aquatica (Ia) showed higher relative µ (MOR) gene expression of 37.75 and 59.76 at 2.878, and 5.813 mg/ml. A combination of Mr and Ia at 1.227, 2.907 and 0.0123, 0.0291 mg/ml extract showed high relative µ (MOR) gene expression at 71.01 and 21.71. These Mitragyna species and the combination (formula details are patent registered), substances and their biological activities can be used for the innovative production of new medicines and further clinical investigation.
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spelling doaj-art-fcabd1ba0ce24de2aee445f7d2cc2d8d2025-08-20T03:13:17ZengNature PortfolioScientific Reports2045-23222025-02-0115111510.1038/s41598-025-89715-5Discovery of rhynchophylline and mitraphylline in two Thai Mitragyna species and the investigation of their biological activity via opioid gene expression analysisRunglawan Sudmoon0Tawatchai Tanee1Warin Wonok2Unchaleeporn Ameamsri3Thomas Liehr4Sakda Daduang5Pornnarong Siripiyasing6Arunrat Chaveerach7Faculty of Law, Khon Kaen UniversityFaculty of Environment and Resource Studies, Mahasarakham UniversityDepartment of Biology, Faculty of Science, Khon Kaen UniversityDepartment of Biology, Faculty of Science, Khon Kaen UniversityInstitute of Human Genetics, Jena University HospitalDivision of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Sciences, Khon Kaen UniversityFaculty of Science and Technology, Rajabhat Mahasarakham UniversityDepartment of Biology, Faculty of Science, Khon Kaen UniversityAbstract Mitragyna speciosa (Ms), M. diversifolia (Md), M. hirsuta (Mh) and M. rotundifolia (Mr) were investigated for phytochemicals by GC-MS and GC-FID, cytotoxicity and genotoxicity testing by MTT and comet assay, and biological activity examination through gene expression of human µ, δ, κ, and nociceptin opioid receptors by qRT-PCR. The opioid substances mitragynine, 7-hydroxymitragynine, and mitraphylline were found in all studied species, and, for first time, rhynchophylline was found in Mr, and mitraphylline in Md, Mh and Mr. The MTT and comet assays of the ethanol and hexane leaf extracts on PBMCs revealed no cytotoxicity and no significant genotoxicity compared to the negative control, except for the hexane leaf extract of Mh, which caused significant DNA damage. The biological activity of the ethanolic extract of the four species showed a binding affinity to the µ (MOR) receptor revealing a relative gene expression of 89.54 and 50.41 by Ms and Md at 1.92 and 1.133 mg/ml, 32.42 and 19.97 mg/ml by Md and Ms at 3.77 and 1.76 mg/ml. Mr contained the three opioids mentioned plus rhynchophylline and showed low relative µ (MOR) gene expression of 16.89 at 0.189 mg/ml, while as an additional species, Ipomoea aquatica (Ia) showed higher relative µ (MOR) gene expression of 37.75 and 59.76 at 2.878, and 5.813 mg/ml. A combination of Mr and Ia at 1.227, 2.907 and 0.0123, 0.0291 mg/ml extract showed high relative µ (MOR) gene expression at 71.01 and 21.71. These Mitragyna species and the combination (formula details are patent registered), substances and their biological activities can be used for the innovative production of new medicines and further clinical investigation.https://doi.org/10.1038/s41598-025-89715-5RhynchophyllineMitraphyllineOpioid receptor geneMitragyna species
spellingShingle Runglawan Sudmoon
Tawatchai Tanee
Warin Wonok
Unchaleeporn Ameamsri
Thomas Liehr
Sakda Daduang
Pornnarong Siripiyasing
Arunrat Chaveerach
Discovery of rhynchophylline and mitraphylline in two Thai Mitragyna species and the investigation of their biological activity via opioid gene expression analysis
Scientific Reports
Rhynchophylline
Mitraphylline
Opioid receptor gene
Mitragyna species
title Discovery of rhynchophylline and mitraphylline in two Thai Mitragyna species and the investigation of their biological activity via opioid gene expression analysis
title_full Discovery of rhynchophylline and mitraphylline in two Thai Mitragyna species and the investigation of their biological activity via opioid gene expression analysis
title_fullStr Discovery of rhynchophylline and mitraphylline in two Thai Mitragyna species and the investigation of their biological activity via opioid gene expression analysis
title_full_unstemmed Discovery of rhynchophylline and mitraphylline in two Thai Mitragyna species and the investigation of their biological activity via opioid gene expression analysis
title_short Discovery of rhynchophylline and mitraphylline in two Thai Mitragyna species and the investigation of their biological activity via opioid gene expression analysis
title_sort discovery of rhynchophylline and mitraphylline in two thai mitragyna species and the investigation of their biological activity via opioid gene expression analysis
topic Rhynchophylline
Mitraphylline
Opioid receptor gene
Mitragyna species
url https://doi.org/10.1038/s41598-025-89715-5
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