Elsevier BVActa Medica Okayama0960-894X962023Direct evaluation of polarity of the ligand binding pocket in retinoid X receptor using a fluorescent solvatochromic agonist129536ENKizukuMiuraFaculty of Pharmaceutical Sciences, Okayama UniversityMichikoFujiharaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMasakiWatanabeDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYutaTakamuraDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMayuKawasakiGraduate School of Integrated Pharmaceutical and Nutritional Sciences, University of ShizuokaShogoNakanoGraduate School of Integrated Pharmaceutical and Nutritional Sciences, University of ShizuokaHirokiKakutaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHigh selectivity of small-molecule drug candidates for their target molecule is important to minimize potential side effects. One factor that contributes to the selectivity is the internal polarity of the ligand-binding pocket (LBP) in the target molecule, but this is difficult to measure. Here, we first confirmed that the retinoid X receptor (RXR) agonist 6-(ethyl(1-isobutyl-2-oxo-4-(trifluoromethyl)-1,2-dihydroquinolin-7-yl)amino)nicotinic acid (NEt-iFQ, 1) exhibits fluorescence solvatochromism, i.e., its Stokes shift depends on the polarity of the solvent, and then we utilized this property to directly measure the internal polarity of the RXRα-LBP. The Stokes shift of 1 when bound to the RXRα-LBP corresponded to that of 1 in chloroform solution. This finding is expected to be helpful for designing RXR-selective ligands. A similar approach should be appliable to evaluate the internal polarity of the LBPs of other receptors.No potential conflict of interest relevant to this article was reported.American Chemical Society (ACS)Acta Medica Okayama0022-26236582022Identification of a Vitamin-D Receptor Antagonist, MeTC7, which Inhibits the Growth of Xenograft and Transgenic Tumors In Vivo60396055ENNegarKhazanWilmot Cancer Institute and Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical CenterKyu KwangKimWilmot Cancer Institute and Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical CenterJeanne N.HansenDepartment of Pediatrics, University of Rochester Medical CenterNiloy A.SinghWilmot Cancer Institute and Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical CenterTaylorMooreWilmot Cancer Institute and Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical CenterCameron W. A.SnyderWilmot Cancer Institute and Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical CenterRavinaPanditaWilmot Cancer Institute and Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical CenterMylaStrawdermanDepartment of Biostatistics and Computational Biology, University of Rochester Medical CenterMichikoFujiharaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYutaTakamuraDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYeJianDivision of Surgery and of Microbiology and Immunology, University of Rochester Medical CenterNicholasBattagliaDivision of Surgery and of Microbiology and Immunology, University of Rochester Medical CenterNaohiroYanoDepartment of Surgery, Division of Surgical Research, Rhode Island Hospital, Alpert Medical School of Brown UniversityYukiTeramotoDepartment of Pathology and Laboratory Medicine, University of Rochester Medical CenterLeggy A.ArnoldDepartment of Chemistry and Biochemistry, University of Wisconsin MilwaukeeRussellHopsonDepartment of Chemistry, Brown UniversityKeshavKishorDepartment of Chemistry, Birla Institute of TechnologySnehaNayakDepartment of Chemistry, Birla Institute of TechnologyDebasmitaOjhaDepartment of Chemistry, Birla Institute of TechnologyAshokeSharonDepartment of Chemistry, Birla Institute of TechnologyJohn M.AshtonGenomics Core Facility, Wilmot Cancer Center, University of Rochester Medical CenterJianWangDepartment of Pharmacology and Department of Biochemistry and Molecular Biology, Penn State College of Medicine, Penn State UniversityMichael T.MilanoDepartment of Radiation Oncology, University of Rochester Medical CenterHiroshiMiyamotoDepartment of Pathology and Laboratory Medicine, University of Rochester Medical CenterDavid C.LinehanDivision of Surgery and of Microbiology and Immunology, University of Rochester Medical CenterScott A.GerberDivision of Surgery and of Microbiology and Immunology, University of Rochester Medical CenterNadaKawarCenter for Breast Health and Gynecologic Oncology, Mercy Medical CenterAjay P.SinghRutgers, The State University of New JerseyErdem D.TabdanovCytoMechanobiology Laboratory, Department of Pharmacology, Penn State College of Medicine, Pennsylvania State UniversityNikolay V.DokholyanDepartment of Pharmacology and Department of Biochemistry and Molecular Biology, Penn State College of Medicine, Penn State UniversityHirokiKakutaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesPeter W.JurutkaSchool of Mathematical and Natural Sciences, Arizona State University, Health Futures CenterNina F.SchorDepartments of Pediatrics, Neurology, and Neuroscience, University of Rochester Medical CenterRachael B.Rowswell-TurnerWilmot Cancer Institute and Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical CenterRakesh K.SinghWilmot Cancer Institute and Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical CenterRichard G.MooreWilmot Cancer Institute and Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical CenterVitamin-D receptor (VDR) mRNA is overexpressed in neuroblastoma and carcinomas of lung, pancreas, and ovaries and predicts poor prognoses. VDR antagonists may be able to inhibit tumors that overexpress VDR. However, the current antagonists are arduous to synthesize and are only partial antagonists, limiting their use. Here, we show that the VDR antagonist MeTC7 (5), which can be synthesized from 7-dehydrocholesterol (6) in two steps, inhibits VDR selectively, suppresses the viability of cancer cell-lines, and reduces the growth of the spontaneous transgenic TH-MYCN neuroblastoma and xenografts in vivo. The VDR selectivity of 5 against RXRα and PPAR-γ was confirmed, and docking studies using VDR-LBD indicated that 5 induces major changes in the binding motifs, which potentially result in VDR antagonistic effects. These data highlight the therapeutic benefits of targeting VDR for the treatment of malignancies and demonstrate the creation of selective VDR antagonists that are easy to synthesize.No potential conflict of interest relevant to this article was reported.Pharmaceutical Society of JapanActa Medica Okayama0009-23637022022Development of Scaled-Up Synthetic Method for Retinoid X Receptor Agonist NEt-3IB Contributing to Sustainable Development Goals146154ENYutaTakamuraDivision of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityKen-ichiMorishitaDivision of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityShotaKikuzawaDivision of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityMasakiWatanabeDivision of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityHirokiKakutaDivision of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama UniversityAbstract
Small-molecular drugs, which are generally inexpensive compared with biopharmaceuticals and can often be taken orally, may contribute to the Sustainable Development Goals (SDGs) adopted by the United Nations. We previously reported the retinoid X receptor (RXR) agonist 4-(ethyl(3-isobutoxy-4-isopropylphenyl)amino)benzoic acid (NEt-3IB, 1) as a small-molecular drug candidate to replace biopharmaceuticals for the treatment of inflammatory bowel disease. The previous synthetic method to 1 required a large amount of organic solvent and extensive purification. In line with the SDGs, we aimed to develop an environmentally friendly, inexpensive method for the large-scale synthesis of 1. The developed method requires only a hydrophobic ether and EtOH as reaction and extraction solvents. The product was purified by recrystallization twice to afford 99% pure 1 at 100 mmol scale in about 30% yield. The optimized process showed a 35-fold improvement of the E-factor (an index of environmental impact) compared to the original method. This work, which changes the solvent used to environmentally preferable ones based on the existing synthetic method for 1, illustrates how synthetic methods for small-molecular drugs can be adapted and improved to contribute to the SDGs.No potential conflict of interest relevant to this article was reported.Frontiers Media SAActa Medica Okayama1663-9812122021A Retinoid X Receptor Agonist Directed to the Large Intestine Ameliorates T-Cell-Mediated Colitis in Mice715752ENRyohtarohMatsumotoDivision of Biochemistry, Graduate School of Pharmaceutical Science and Faculty of Pharmacy, Keio UniversityDaisukeTakahashiDivision of Biochemistry, Graduate School of Pharmaceutical Science and Faculty of Pharmacy, Keio UniversityMasakiWatanabeDivision of Pharmaceutical Sciences, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama UniversityShunsukeNakataniDivision of Pharmaceutical Sciences, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama UniversityYutaTakamuraDivision of Pharmaceutical Sciences, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama UniversityYujiKurosakiDivision of Pharmaceutical Sciences, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama UniversityHirokiKakutaDivision of Pharmaceutical Sciences, Graduate School of Medicine Dentistry and Pharmaceutical Sciences, Okayama UniversityKojiHaseDivision of Biochemistry, Graduate School of Pharmaceutical Science and Faculty of Pharmacy, Keio UniversityRetinoid X receptor (RXR) is a nuclear receptor that heterodimerizes with several nuclear receptors, integrating ligand-mediated signals across the heterodimers. Synthetic RXR agonists have been developed to cure certain inflammatory diseases, including inflammatory bowel diseases (IBDs). However, pre-existing RXR agonists, which are lipophilic and readily absorbed in the upper intestine, cause considerable adverse effects such as hepatomegaly, hyperlipidemia, and hypothyroidism. To minimize these adverse effects, we have developed an RXR agonist, NEt-3IB, which has lipophilic and thus poorly absorptive properties. In this study, we evaluated the effects of NEt-3IB in an experimental murine colitis model induced through the adoptive transfer of CD45RB(high)CD4(+) T cells. Pharmacokinetic studies demonstrated that the major portion of NEt-3IB was successfully delivered to the large intestine after oral administration. Notably, NEt-3IB treatment suppressed the development of T cell-mediated chronic colitis, as indicated by improvement of wasting symptoms, inflammatory infiltration, and mucosal hyperplasia. The protective effect of NEt-3IB was mediated by the suppression of IFN-gamma-producing Th1 cell expansion in the colon. In conclusion, NEt-3IB, a large intestine-directed RXR agonist, is a promising drug candidate for IBDs.No potential conflict of interest relevant to this article was reported.MDPIActa Medica Okayama2072-66941262020Paclitaxel and Sorafenib: The Effective Combination of Suppressing the Self-Renewal of Cancer Stem Cells1360ENHend M.NawaraDepartment of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama UniversitySaid M.AfifyLaboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama UniversityGhmkinHassanLaboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama UniversityMaram H.ZahraLaboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama UniversityMarwa N.AtallahVertebrates Embryology and Comparative Anatomy, Zoology Department, Faculty of Science, Menoufia UniversityHagerMansourDepartment of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama UniversityHagar A.Abu QuoraLaboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama UniversityMd JahangirAlamDepartment of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama UniversityAmiraOsmanLaboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama UniversityHirokiKakutaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHirokiHamadaDepartment of Life Science, Faculty of Science, Okayama University of ScienceAkimasaSenoLaboratory of Nano-Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama UniversityMasaharuSenoDepartment of Medical Bioengineering, Graduate School of Natural Science and Technology, Okayama UniversityCombination therapy, which is a treatment modality combining two or more therapeutic agents, is considered a cornerstone of cancer therapy. The combination of anticancer drugs, of which functions are different from the other, enhances the efficiency compared to the monotherapy because it targets cancer cells in a synergistic or an additive manner. In this study, the combination of paclitaxel and sorafenib in low concentration was evaluated to target cancer stem cells, miPS-BT549cmP and miPS-Huh7cmP cells, developed from mouse induced pluripotent stem cells. The synergistic effect of paclitaxel and sorafenib on cancer stem cells was assessed by the inhibition of proliferation, self-renewal, colony formation, and differentiation. While the IC(50)values of paclitaxel and sorafenib were approximately ranging between 250 and 300 nM and between 6.5 and 8 mu M, respectively, IC(50)of paclitaxel reduced to 20 and 25 nM, which was not toxic in a single dose, in the presence of 1 mu M sorafenib, which was not toxic to the cells. Then, the synergistic effect was further assessed for the potential of self-renewal of cancer stem cells by sphere formation ability. As a result, 1 mu M of sorafenib significantly enhanced the effect of paclitaxel to suppress the number of spheres. Simultaneously, paclitaxel ranging in 1 to 4 nM significantly suppressed not only the colony formation but also the tube formation of the cancer stem cells in the presence of 1 mu M sorafenib. These results suggest the combination therapy of paclitaxel and sorafenib in low doses should be an attractive approach to target cancer stem cells with fewer side effects.No potential conflict of interest relevant to this article was reported.Pharmaceutical Society of Japan Acta Medica Okayama0031-690313962019研究公正化，医薬品開発のスピードアップのための「信頼性確保の知識・方法論」の産学官での共有を目指して873874ENHirokiKakutaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHirokazuSudoNo potential conflict of interest relevant to this article was reported.Pharmaceutical Society of JapanActa Medica Okayama0031-690313962019アカデミアにおける実験記録の悩み それを解決するには887890ENHirokiKakutaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences There is currently a major effort to promote drug discovery in academia as a way to seed new drug development in the pharmaceutical industry. However, there are concerns in industry about the quality of drug candidates generated in academic institutions. These concerns encompass culture and perceptions with respect to intellectual property management, the process of product development, and the reliability of scientific data. Questions about data reliability underscore the particularly serious problem of mistrust in academic research. Therefore, the author became interested in the topic of industry standards for quality assurance (QA) and arranged training workshops at Okayama University on the appropriate methods for recording experimental notes by lecturers involved in QA. The outcomes are presented here.No potential conflict of interest relevant to this article was reported.American Chemical SocietyActa Medica Okayama0022262362192019Competitive Binding Assay with an Umbelliferone-Based Fluorescent Rexinoid for Retinoid X Receptor Ligand Screening88098818ENShoyaYamadaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMayuKawasakiMichikoFujiharaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMasakiWatanabeDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYutaTakamuraDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMahoTakiokuDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesHiromiNishiokaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesYasuoTakeuchiDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesMakotoMakishimaDivision of Biochemistry, Department of Biomedical Sciences, Nihon University School of MedicineTomoharuMotoyamaGraduate School of Integrated Pharmaceutical and Nutritional Sciences, University of ShizuokaSoheiItoGraduate School of Integrated Pharmaceutical and Nutritional Sciences, University of ShizuokaHiroakiTokiwaDepartment of Chemistry and Research Center of Smart Molecules, Rikkyo UniversityShogoNakanoGraduate School of Integrated Pharmaceutical and Nutritional Sciences, University of ShizuokaHirokiKakutaDivision of Pharmaceutical Sciences, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences Ligands for retinoid X receptors (RXRs), "rexinoids", are attracting interest as candidates for therapy of type 2 diabetes and Alzheimer's and Parkinson's diseases. However, current screening methods for rexinoids are slow and require special apparatus or facilities. Here, we created 7-hydroxy-2-oxo-6-(3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydronaphthalen-2-yl)-2H-chromene-3-carboxylic acid (10, CU-6PMN) as a new fluorescent RXR agonist and developed a screening system of rexinoids using 10. Compound 10 was designed based on the fact that umbelliferone emits strong fluorescence in a hydrophilic environment, but the fluorescence intensity decreases in hydrophobic environments such as the interior of proteins. The developed assay using 10 enabled screening of rexinoids to be performed easily within a few hours by monitoring changes of fluorescence intensity with widely available fluorescence microplate readers, without the need for processes such as filtration.No potential conflict of interest relevant to this article was reported.