Kyushu University Academic Staff Educational and Research Activities Database
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kuniyoshi shimizu Last modified date:2020.06.15

Associate Professor / Systematic Forest and Forest Products Science
Department of Agro-environmental Sciences
Faculty of Agriculture


Graduate School


E-Mail
Homepage
https://kyushu-u.pure.elsevier.com/en/persons/kuniyoshi-shimizu
 Reseacher Profiling Tool Kyushu University Pure
http://ffpsc.agr.kyushu-u.ac.jp/sffps/   
Homepage of Laboratory of Systematic Forest and Forest Products Sciences.
http://shinrinken.main.jp/ .
Phone
092-642-3002
Fax
092-642-3002
Academic Degree
PhD
Country of degree conferring institution (Overseas)
No
Field of Specialization
Natural Products Bioorganic Chemistry
Total Priod of education and research career in the foreign country
02years00months
Outline Activities
The investigation of LOHAS-oriented biological activity of forest products and their applications

Forest has been known to have healing power for mind and body. Our group deals with LOHAS (Life Of Health And Sustainability) oriented biological aspects of forest products and their mode of action. In most cases this involves investigating the biological activity of extractives of mushroom , woods and plant etc, such as anti-androgenic activity, prevention effect of bone loss in calcium deficient ovariectomized rats of mushroom, healing aroma, and melanin biosynthesis inhibitory activity of wood extractives. Our research cover several practical fields such as cosmetics, aroma, functional food, living environment, toiletry etc.

1, Search for biologically active substance from plants.
2, Search for biologically active substance from mushrooms.
3, Studies on the mode of action of bioactive natural substances and their structural criteria.
4, The physiological and psychological effect of aroma (natural volatiles) on human.
5, Development of bioactive substances and extracts for practical use (cosmetics, aroma, functional food, toiletry and living environment.)
Research
Research Interests
  • In vitro functional evaluation test of ethanol and water extract of Houttuynia cordata
    keyword : Houttuynia cordata
    2019.09~2020.02.
  • A supplement containing Nejimebiwa tea as a functionally labeled food that contributes to the suppression of allergic symptoms
    keyword : Eriobotrya japonica
    2019.07~2020.02.
  • Verification of effects on human cognitive function using foods containing peanut extract
    keyword : Arachis hypogaea
    2019.09~2020.03.
  • Basic research on improvement of cognitive function of ingredients containing Yamabushitake
    keyword : Hericium erinaceus
    2019.04~2020.03.
  • Volatile compounds emitted from Juncus effusus and their functions
    keyword : Juncus effusus, Igusa,GC/MS,Tatami
    2018.04~2021.03.
  • Functional components isolated from Olive
    keyword : Olive
    2013.04.
  • Comparison of the physiological and psychological response of the component profile and human essential oil aroma prepared from cedar leaves and wood parts
    keyword : Cryptomeria japonica essential oil
    2013.04~2015.02.
  • Wild Mushroom in Nepal: Some potential Candidates as Antioxidant and ACE Inhibition Sources
    keyword : Wild Mushroom in Nepal
    2013.04.
  • Comparative Biological Study of Roots, Stems, Leaves, and Seeds of
    Angelica shikokiana Makino

    keyword : Angelica shikokiana
    2013.04.
  • Search for bioactive compound from the waste of Ejyptian plants
    keyword : Medicinal Plants from Ejypt
    2013.04.
  • HMGCoA reductase inhibitors from natural resources
    keyword : HMGCoA reductase inhibitor
    2008.05~2010.03.
  • Search for hair growth agents from natural products
    keyword : hair growth
    2008.05~2010.03.
  • Functional components isolated from Eriobotrya japonica
    keyword : Eriobotrya japonica
    2008.05.
  • Search for bioactive compounds from the bamboo and the skin of bamboo shoot
    keyword : bamboo shoot, biological activity
    2007.05.
  • Volatile compounds emitted from woods and their functions
    keyword : terpene, volatile
    2006.04.
  • The physiological and psychological effects of volatiles emitted from natural resources on human
    keyword : natural product, volatile, aroma, essential oil
    2005.04.
  • The aldose reductase inhibitors from Ganoderma lucidum
    keyword : aldose reductase, Ganoderma lucidum
    2004.04The inhibitory activity of the ethanol extract of Ganoderma lucidum on aldose reductase..
  • Search for the preventive compounds from mushroom on benign prostate hyperplasia
    keyword : Ganoderma lucidum, fruiting body, antiandrogen, 5a-reductase
    2001.04Screening of the inhibitors of benign prostatic hyperplasia from edible and medicinal mushrooms..
  • Search for the compound with estrogenic activity from mushrooms
    keyword : fruiting body, estrogenic activity, Pleurotus eryngii, Ganoderma lucidum
    2002.04Screening of the prophylactic compounds for osteoporosis from edible and medicinal mushrooms..
Academic Activities
Books
1. I. Kubo, T. J. Ha, K. Shimizu, Molecular Design of Soybean Lipoxygenase Inhibitors Based on Natural Products, A Comprehensive Survey of International Soybean Research - Genetics, Physiology, Agronomy and Nitrogen Relationships, InTech, James E. Board (Ed.)
DOI: 10.5772/52703., 2013.03, [URL].
2. N. Masuoka, K. Shimizu, I. Kubo, Antioxidant Activity of Anacardic Acids
In: Antioxidants and Biocides from Latin American Plants;
, CABI Publishing (UK), C.L. Cespedes, D.A. Sampietro, D.S. Seigler, Ed., 2012.11.
3. I. Kubo, K. Fujita, K. Shimizu, Anti-Salmonella Agents from Tanacetum Balsamita, A Brazilian Medicinal Plant, and their Application
In: Antioxidants and Biocides from Latin American Plants
, CABI Publishing (UK), C.L. Cespedes, D.A. Sampietro, D.S. Seigler, Ed., 2012.11.
4. I. Kubo, K. Shimizu, K. Nihei, MANICOBA, A quercetin rich Amazonian dish
In: Quercetin: Dietary Sources, Functions and Health Benefits
, Nova Science Publishers, Inc., 2012.11.
5. I. Kubo, K. Shimizu, K. Fujita, Naturally Occurring Antifungal Agents and Their Modes of Action.
Fungicides for Plant and Animal Diseases
, InTech, D. Dhanasekaran, N. Thajuddin and A. Panneerselvam (Ed.), 2012.11, [URL].
Reports
1. Amira Mira, kuniyoshi shimizu, An Update on Antitumor Activity of Angelica Species, Medicinal Plants - Recent Advances in Research and Development, pp 363-371, 2016.10.
2. I. Kubo, T. J. Ha, K. Shimizu, Molecular Design of Soybean Lipoxygenase Inhibitors Based on Natural Products, A Comprehensive Survey of International Soybean Research - Genetics, Physiology, Agronomy and Nitrogen Relationships, James E. Board (Ed.), ISBN: 978-953-51-0876-4, InTech, DOI: 10.5772/52703., 2013.01.
3. N. Masuoka, K. Shimizu, I. Kubo, Antioxidant Activity of Anacardic Acids
, In: Antioxidants and Biocides from Latin American Plants; C.L. Cespedes, D.A. Sampietro, D.S. Seigler, Ed., CABI Publishing (UK)(2012), In: Antioxidants and Biocides from Latin American Plants;
C.L. Cespedes, D.A. Sampietro, D.S. Seigler, Ed., CABI Publishing (UK)(2012), 2012.02.
4. I. Kubo, K. Fujita, K. Shimizu, Anti-Salmonella Agents from Tanacetum Balsamita, A Brazilian Medicinal Plant, and their Application, In: Antioxidants and Biocides from Latin American Plants; C.L. Cespedes, D.A. Sampietro, D.S. Seigler, Ed., CABI Publishing (UK), 2012.02.
5. I. Kubo, K. Shimizu, K. Nihei, MANICOBA, A quercetin rich Amazonian dish, In: Quercetin: Dietary Sources, Functions and Health Benefits- Nova Science Publishers, Inc., 2012.02.
6. I. Kubo, K. Shimizu, K. Fujita, Naturally Occurring Antifungal Agents and Their Modes of Action., In : Fungicides Volume I; D. Dhanasekaran, N. Thajuddin, A. Panneerselvam, Ed., InTech - Open Access Publisher, 2012.02.
7. I. Kubo, N. Masuoka, T. J. Ha, K. Shimizu, K. Nihei, Multifunctional Antioxidant Activities of Alkyl Gallates, The Open Bioactive Compounds Journal, 3, 1-11(2010), 2010.06.
8. K. Shimizu, R. Kondo, K. Sakai, Tyrosinase inhibitors from Artocarpus incisus., Research Advances in Phytochemistry, 1, p19-34, 2000.01.
9. K. Shimizu, Tyrosinase and 5a-Reductase Inhibitory Components from Artocarpus incisus, Bull. Kyushu Univ. For., 82, p21-63, 2001.01.
10. I. Kubo, T. J. Ha, T. Nitoda, K. Shimizu, A. Kubo, Multifunctional Acne Control Agents , Current Topics in Phytochemistry, 9, 95-102 (2008) , 2008.06.
Papers
1. Seiko Goto, Hinako Suzuki, Toshinori Nakagawa, Kuniyoshi Shimizu, The Effect of Eucalyptol on Nursing Home Residents, SCIENTIFIC REPORTS, 2020.03, Eucalyptol is one of the most popular volatile components. It is used in many essential oils for relieving
sinus and lung congestion caused by a variety of conditions. This pilot study sought to analyze clinical
evidence for the effect of the scent of eucalyptol on the cognitive function of elderly people. Seventy
nursing-home residents with cognitive impairment were recruited. Three one-week experiments were
performed: eucalyptol scent was diffused in bedrooms with a diffuser only at wake-up time in the first
experiment, and at wake-up time and bedtime in the second and third experiments. Results showed
that although an improvement was not seen when using Mini Mental State Examination (MMSE) and
Cohen-Mansfild Agitation Inventory (CMAI) measures, Dementia Behavior Disturbance Scale (DBD)
scores improved significantly, even though no subject reported perceiving the scent. The significant
improvements of the behaviour were found not only among the subjects whose room had a diffuser but
also among the subjects who were exposed to an unperceivable level of eucalyptol drifted in the living
room..
2. Fahd M. Abdelkarem, Ezz-Eldin K. Desoky, Alaa M. Nafady, Ahmed E. Allam, Aldoushy Mahdy, Maiki Nagata, Tomofumi Miyamoto, Kuniyoshi Shimizu., Isolation of new secondary metabolites from gorgonian soft coral Heteroxenia fuscescens collected from Red Sea, Phytochemistry Letters.
3. Tukiran, Andika Pramudya Wardana, Nurul Hidajati, Kuniyoshi Shimizu, Chemical components and antioxidant activities of methanol extract of Syzygium polycephalum Miq. stem bark (Myrtaceae), Indian Journal of Natural Products and Resources , 10, (2), 127-136, 2019.06, This study aimed to reveal chemical components and screen antioxidant activity of methanolic extract of Syzygium
polycephalum stem bark. The chemical components were isolated by using column chromatographic techniques and
determined by spectroscopic methods (UV-Vis, FTIR, MS, and NMR) and comparison with literature data, in which the
antioxidant activity was performed using 2,2’-diphenyl-1-picrylhydrazyl (DPPH). The present investigation resulted in four
compounds involving gallic acid, pinostrobin, 3,4,3’-tri-O-methylellagic acid, and 3,3’-di-O-methylellagic acid. The four
compounds were all in the form of phenolic compounds found from the extract whereas 3,4,3’-tri-O-methylellagic acid and
3,3’-di-O-methylellagic acid were ellagic acid derivatives. The methanolic extract, the four compounds, and vitamin C (as
positive control) showed antioxidant activity against DPPH with the IC50 value of 99.9; 10.0; 183.2; 72.1; 63.3; and 13.9
µg/mL, respectively. The antioxidant activity of gallic acid was more active than that of vitamin C. The present study
confirms that Syzygium polycephalumis rich in phenolic compounds and natural antioxidants.
.
4. Tukiran,A. P. Wardana, N. Hidayati, K. Shimizu, An Ellagic Acid Derivative and Its Antioxidant Activity of Chloroform Extract of Stem Bark of Syzygium polycephalumMiq. (Myrtaceae), Indones. J. Chem, 18, 1, 26-34, 2018.01, The investigation of the Syzygium polycephalum Miq. (Myrtaceae) aimed to assess the phytochemical contents and antioxidant activity of the chloroform fraction. In this study, the fraction was obtained from methanol extract of S. polycephalum stem bark partitionated by chloroform. An ellagic acid derivative was successively isolated from the chloroform fraction. The molecular structure of isolated compound was elucidated and established as 3,4,3’-tri-O-methylellagic acid through extensive spectroscopic studies including UV-Vis, FTIR, NMR and LC-MS analyses and by comparison with literature data. The finding of the isolated compound is the first time from the plant, although the isolated compound previously have been found in the other Syzygium species such as S. cumini together with ellagic acid and 3,3’-di-O-methylellagic acid. The chloroform fraction, isolated compound, and vitamin C showed antioxidant activity against 2,2’-diphenyl-1-picrylhydrazyl (DPPH) with IC50 value of 163.6, 72.1, and 11.5 μg/mL, respectively..
5. Yhiya Amen, Enos Tangke Arung, Mohamed S. Afifi, Ahmed F. Halim, Ahmed Ashour, Ryoji Fujimoto, Takahiro Goto, Kuniyoshi Shimizu, Melanogenesis inhibitors from Coix lacryma-jobi seeds in B16-F10 melanoma cells, Natural Product Research, 10.1080/14786419.2017.1292270, 31, 23, 2712-2718, 2017.12, An adenine derivative, 9-β-D-glucopyranosyl adenine, reported for the first time from a natural source, in addition to nine known compounds were isolated from the seeds of Coix lacryma-jobi. Their structures were elucidated based on extensive spectroscopic and chemical studies. The isolated compounds and the ethanol extract have been assayed for melanin inhibition using B16-F10 melanoma cell line. The results of our study suggested the potential use of Coix lacryma-jobi seeds as a skin whitening agent and reveal the seeds to be a rich source of important phytochemicals with melanogenesis inhibitory activity. Among the isolated compounds, coixol (2) and 2-O-β-glucopyranosyl-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (8) exhibited potent melanogenesis inhibitory activity with no obvious melanocytotoxicity. The rest of the compounds showed weak to moderate activity..
6. Fawzia Ibrahim, Asmaa Kamal El-Deen, Samah Abo El Abass, Kuniyoshi Shimizu, An ecofriendly green liquid chromatographic method for simultaneous determination of nicotinamide and clindamycin phosphate in pharmaceutical gel for acne treatment, Journal of Food and Drug Analysis, 10.1016/j.jfda.2016.09.009, 25, 3, 741-747, 2017.07, A new green micellar liquid chromatographic method was developed and validated for the quantitative estimation of nicotinamide (NICO) and clindamycin phosphate (CLD) in bulk and pharmaceutical gel formulation. The analytes are well resolved in less than 6.0 minutes using micellar mobile phase consisting of 0.10M sodium dodecyl sulfate (SDS), 0.3% triethylamine, and 10% 2-propanol in 0.02M orthophosphoric acid at pH 3.0, running through an Eclipse XDB-C8 column (150 mm × 4.6 mm, 5 μm particle size) with flow rate 1.0 mL/min. The effluent was monitored with diode array detection at 210 nm. The retention times of NICO and CLD were 3.8 minutes and 5.6 minutes, respectively. The method was validated according to the International Conference on Harmonisation (ICH) guidelines in terms of linearity, limit of detection, limit of quantification, accuracy, precision, robustness, and specificity to prove its reliability. Linear correlation was achieved by plotting the peak area of each drug against its concentration. It was found to be rectilinear in the ranges of 1.0–40.0 μg/mL and 0.5–15.0 μg/mL with limits of detection of 0.06 μg/mL and 0.03 μg/mL and limits of quantification of 0.19 μg/mL and 0.09 μg/mL for NICO and CLD, respectively. The method was successfully implemented for the simultaneous determination of the analytes in their bulk powder and combined gel formulation with high % recoveries. The ease of sample treatment facilitates and greatly expedites the treatment with reduced cost and improved accuracy of the procedure..
7. Yhiya Amen, Qinchang Zhu, Hai-Bang Tran, Mohamed S. Afifi, Ahmed F. Halim,, Ahmed Ashour, Kuniyoshi Shimizu, Partial contribution of Rho-kinase inhibition to the bioactivity of Ganoderma lingzhi and its isolated compounds; Insights on discovery of natural Rho-kinase inhibitors, Journal of Natural Medicines, 71, 2, 380-388, 2017.01, Recent studies identified Rho-kinase enzymes (ROCK-I and ROCK-II) as important targets that are involved in a variety of diseases. Synthetic Rho-kinase inhibitors have emerged as potential therapeutic agents to treat disorders such as hypertension, stroke, cancer, diabetes, glaucoma, etc. Our study is the first to screen the total ethanol extract of the medicinal mushroom Ganoderma lingzhi with thirty-five compounds for Rho-kinase inhibitory activity. Moreover, a molecular binding experiment was designed to investigate the binding affinity of the compounds at the active sites of Rho-kinase enzymes. The structure-activity relationship analysis was investigated. Our results suggest that the traditional uses of G. lingzhi might be in part due to the ROCK-I and ROCK-II inhibitory potential of this mushroom. Structure-activity relationship studies revealed some interesting features of the lanostane triterpenes that potentiate their Rho-kinase inhibition. These findings would be helpful for further studies on the design of Rho-kinase inhibitors from natural sources and open the door for contributions from other researchers for optimizing the development of natural Rho-kinase inhibitors..
8. Fawzia Ibrahim, Mohie K.Sharaf El-Din, Asmaa Kamal El-Deen, Kuniyoshi Shimizu, Micellar HPLC method for simultaneous determination of ethamsylate and mefenamic acid in presence of their main impurities and degradation products, Journal of Chromatographic Science, 10.1093/chromsci/bmw143, 55, 1, 23-29, 2017.01, An eco-friendly sensitive, rapid and less hazardous micellar liquid chromatographic method was developed and validated for the simultaneous analysis of ethamsylate (ETM) and mefenamic acid (MFA) in the presence of hydroquinone (HQ) and 2,3-dimethylaniline (DMA) the main impurities of ETM and MFA, respectively. Good chromatographic separation was attained using Eclipse XDBC8 column (150mm × 4.6 mm, 5 μm particle size) adopting UV detection at 300 nm with micellar mobile phase consisting of 0.12M sodium dodecyl sulfate, 0.3% triethylamine and 15% 2- propanol in 0.02M orthophosphoric acid (pH 7.0) at 1.0 mL/min. The analytes were well resolved in <6.0 min, ETM (tR = 1.55 min), HQ (tR = 1.95 min), MFA (tR = 4.55 min) and DMA (tR = 5.80 min). Different validation parameters were examined as recommended by international conference on harmonization (ICH) guidelines. The method was linear over the concentration ranges of 0.5-18.0, 0.5-20.0, 0.01-0.5 and 0.02-0.2 μg/mL with limits of detection of 0.118, 0.159, 0.005 and 0.005 μg/ mL and limits of quantification of 0.358, 0.482, 0.014 and 0.015 μg/mL for ETM, MFA, HQ and DMA, respectively. The suggested method was successfully applied for the determination of the two drugs in their bulk powder, laboratory-prepared mixtures, single-ingredient and co-formulated tablets. The obtained results were in accordance with those of the comparison method. The method can also detect trace amounts of HQ and DMA as the main impurities of ETM and MFA, respectively, within the BP limit (0.1%) for both impurities. Furthermore, it is a stability-indicating one for the determination of ETM in its pure form, single-component tablet and co-formulated tablets with other drugs..
9. Tukiran, F.Mahmudah, N. Hidayati, K.Shimizu, A phenolic acid and its antioxidant activity from stem bark of chloroform fraction of Syzygium littorale (Blume) amshoff (Myrtaceae), Molekul, 11, 2, 180-189, 2016.11, This study describes the chemical investigations of Syzygiumlittorale (S.littorale), a plant
belonging to the family Myrtaceae, and their antioxidant properties. As far, there is no study due to
chemical investigations conducted to the plant either on leaves, fruits, stem bark or the others. The
stem bark of S. littorale was extracted with organic solvent and the extracts were then fractionated
and purified by using standard chromatographic techniques. The molecular structure of pure
isolated pure compound was elucidated and established through extensive spectroscopic studies
and by comparing with literature data and authentic sample. The dried and powdered stem bark of
S. littorale was extracted with methanol and then partitioned by chloroform. Chromatographic
(vacuum liquid chromatography and gravitational column chromatography) fractionation and
purification of the chloroform soluble fraction that is always monitored by TLC yielded a pure
isolated pure compound. The purified compound was then identified and justified as a phenolic
acid i.e. gallic acid. This compound is found for the first time from this plant, although it has
previously been found in other Syzygium species such as S. aromaticum, S. cumini, S. polyanthum,
S. cordatum, etc. The chloroform fraction, isolated compound, and vitamin C showed very strong
antioxidant activity against 2,2‟-diphenyl-1-picrylhydrazyl (DPPH) with the IC50 value of 23.2; 7.5;
and 12.5 ug/mL, respectively..
10. F.Ibrahim, A.K.El-Deen, S.A.El Abass, Shimizu K, An Ecofriendlly Green Liquid Chromatographic Method for Simultaneous Determination of Nicotinamide and Clindamycin phosphate in Pharmaceutical Gel for Acne Treatment, Jounal of Food and Drug Analysis, 25, 3, 741-747, 2016.09, A new green micellar liquid chromatographic method was developed and validated for the quantitative estimation of nicotinamide (NICO) and clindamycin phosphate (CLD) in bulk and pharmaceutical gel formulation. The analytes are well resolved in less than 6.0 minutes using micellar mobile phase consisting of 0.10M sodium dodecyl sulfate (SDS), 0.3% triethylamine, and 10% 2-propanol in 0.02M orthophosphoric acid at pH 3.0, running through an Eclipse XDB-C8 column (150 mm×4.6 mm, 5 μm particle size) with flow rate 1.0 mL/min. The effluent was monitored with diode array detection at 210 nm. The retention times of NICO and CLD were 3.8 minutes and 5.6 minutes, respectively. The method was validated according to the International Conference on Harmonisation (ICH) guidelines in terms of linearity, limit of detection, limit of quantification, accuracy, precision, robustness, and specificity to prove its reliability. Linear correlation was achieved by plotting the peak area of each drug against its concentration. It was found to be rectilinear in the ranges of 1.0-40.0 μg/mL and 0.5-15.0 μg/mL with limits of detection of 0.06 μg/mL and 0.03 μg/mL and limits of quantification of 0.19 μg/mL and 0.09 μg/mL for NICO and CLD, respectively. The method was successfully implemented for the simultaneous determination of the analytes in their bulk powder and combined gel formulation with high % recoveries. The ease of sample treatment facilitates and greatly expedites the treatment with reduced cost and improved accuracy of the procedure..
11. Y.M.Amen, Q.Zhu, M.S.Afifi, A.Ashour, Shimizu K, New cytotoxic lanostanoid triterpenes from Ganoderma lingzhi, Phytochemistry Letters, 17, 64-70, 2016.09, Further chemical investigation of the metabolites in the fruiting bodies of Ganoderma lingzhi resulted in isolation of eight triterpenes; two of them are new triterpene acid ethyl esters. Their structures were established based on spectroscopic studies and comparison with the known related compounds. The anticancer potential of the isolates were tested with an in vitro cytotoxic assay against five human cancer cell lines (MCF-7, HeLa, HCT-116, Caco-2 and HepG2) and two normal human cell lines (TIG-1 and HF19). Results showed that the new compounds have a strong to moderate selective cytotoxic activity against MCF-7 while they showed moderate to weak activity against HeLa cell line. Potent cytotoxic activities of some of the known isolated compounds are reported for the first time..
12. A.Sallam, A.Mira, A.Ashour, Shimizu K, Acetylcholine esterase inhibitors and melanin synthesis inhibitors from Salvia officinalis, Phytomedicine, 23, 10, 1005-1011, 2016.09, BACKGROUND:
Salvia officinalis is a traditionally used herb with a wide range of medicinal applications. Many phytoconstituents have been isolated from S. officinalis, mainly phenolic diterpenes, which possess many biological activities.

PURPOSE:
This study aimed to evaluate the ability of the phenolic diterpenes of S. officinalis to inhibit acetylcholine esterase (AChE) as well as their ability to inhibit melanin biosynthesis in B16 melanoma cells.

METHODS:
The phenolic diterpenes isolated from the aerial parts of S. officinalis were tested for their effect on melanin biosynthesis in B16 melanoma cell lines. They were also tested for their ability to inhibit AChE using Ellman's method. Moreover, a molecular docking experiment was used to investigate the binding affinity of the isolated phenolic diterpenes to the amino acid residues at the active sites of AChE.

RESULTS:
Seven phenolic diterpenes-sageone, 12-methylcarnosol, carnosol, 7b-methoxyrosmanol, 7a-methoxyrosmanol, isorosmanol and epirosmanol-were isolated from the methanolic extract of the aerial parts of S. officinalis. Isorosmanol showed a melanin-inhibiting activity as potent as that of arbutin. Compounds 7a-methoxyrosmanol and isorosmanol inhibited AChE activity by 50% and 65%, respectively, at a concentration of 500 µM.

CONCLUSIONS:
The results suggest that isorosmanol is a promising natural compound for further studies on development of new medications which might be useful in ageing disorders such as the declining of cognitive functions and hyperpigmentation..
13. T. Kohno, T. Hai-Bang, Q. Zhu, S. Sakamoto, H. Tanaka, S. Morimoto, Shimizu K, Tubulin polymerization stimulating-activity of Ganoderma triterpenoids., J Nat Med, 71, 2, 457-462, 2017.04, Tubulin polymerization is an important target for anticancer therapies. Even though the potential of Ganoderma triterpenoids against various cancer targets had been well documented, studies on their tubulin polymerization-stimulating activity are scarce. This study was conducted to evaluate the effect of Ganoderma triterpenoids on tubulin polymerization. A total of twenty-four compounds were investigated using an in vitro tubulin polymerization assay. Results showed that most of the studied triterpenoids exhibited microtuble-stabilizing activity to different degrees. Among the investigated compounds, ganoderic acid T-Q, ganoderiol F, ganoderic acid S, ganodermanontriol and ganoderic acid TR were found to have the highest activities. A structure-activity relationship (SAR) analysis was performed. Extensive investigation of the SAR suggests the favorable structural features for the tubulin polymerization-stimulating activity of lanostane triterpenes. These findings would be helpful for further studies on the potential mechanisms of the anticancer activity of Ganoderma triterpenoids and give some indications on the design of tubulin-targeting anticancer agents..
14. Horiba Hiroki,Nakagawa Toshinori,Zhu Qinchang,Ashour Ahmed,Watanabe Atsushi, Shimizu Kuniyoshi, Biological activities of extracts from different parts of cryptomeria japonica, Natural Product Communications, 11, 9, 1337-1342, 2016.09, Cryptomeria japonica is one of the most important forest tree species in Japan. To increase the demand for domestic timber, broad uses related to the functional attributes of C. japonica must be developed. Several studies have examined the usefulness of C. japonica in terms of its biological activities, but a comprehensive study subjecting all parts of the C. japonica plant to the same solvent for extraction has not been done. Here, methanol extractions from the leaves, branches, bearing branches, male flowers, female flowers, cones, bark, heartwood, sapwood, pith, rhizomes, roots, and pollen were subjected to several in vitro assays of their biological activities such as antioxidant activity, anti-lipase activity, antibacterial activity and melanin-biosynthesis-inhibition activity. Their total phenolic content was also determined. The methanol extracts from each part of C. japonica except for pollen showed strong activities in the bioactivity assays. Furthermore, the methanol extracts were analyzed by GC/MS. The phytochemical profile varied among extracts from various parts of C. japonica. Our results suggest the great potential of C. japonica for use as a functional ingredient in health-related products..
15. F. Ibrahim, M. K. S. El-Din, R. N. El-Shaheny, A. K. El-Deen, Shimizu K, Simultaneous determination of four vasoactive phytochemicals in different pharmaceutical preparations by a simple HPLC-DAD method, Analytical Methods, 10.1039/C5AY03003J, 8, 1858-1866, 2016.02, A new, simple isocratic HPLC method was developed and validated for the simultaneous estimation of four vasoactive phytochemicals: ascorbic acid (ASC), rutin (RUT), hesperidin (HSP), and diosmin (DSM) in different pharmaceutical preparations. Separation of the four compounds was achieved in about 8 min on a phenomenex-C18 column (250 mm x 4.6 mm, 5 μm particle size) using a mobile phase consists of 50 mM phosphate buffer (pH 4.0):acetonitrile (75:25, v/v) eluted at 1.0 mL min-1 with diode array detection at 275 nm. Linear correlation was obeyed over the concentration ranges of 2.0-60.0, 1.0-20.0, 2.0-50.0, and 5.0-100.0 μg mL-1 for ASC, RUT, HSP, and DSM, respectively. The developed method is efficient, reproducible and appropriate as a versatile method for quality control of various combinations of these drugs in different pharmaceutical preparations with high values of %recoveries which agreed well with the labeled claim. The method is also a stability-indicating one for DSM in its pure state and single-component tablets. A comparison of the results obtained by the developed method with those of the comparison methods confirmed no significant differences between them with respect to accuracy and precision..
Presentations
1. Taisuke Nakashima, Akiyoshi Honden, Minkai Sun, Yuri Yoshimura, Toshinori Nakagawa, Hiroya Ishikawa, Jun Nagano, Yuki Yamada, Tsuyoshi Okamoto, Yuichiro Watanabe, Shinji Yasunari, Koichiro Ohnuki, Noboru Fujimoto, Kuniyoshi Shimizu. , Effects of Japanese cedar scent on the human brain function, Asian Natural Product Conference (Q-AOS), 2020.05.
2. Rogers Mwakalukwa, Kuniyoshi Shimizu, Cultivar-specific metabolic profiling of olive leaves cultivars by LCMS based non-targeted metabolomics for the determination of AChE inhibitor metabolites , Asian Natural Product Conference (Q-AOS), 2020.05.
3. Yunfei XIAO,Yhiya AMEN,Asmaa KAMAL EL-DEEN,Naomichi TAKEMOTO,Dongmei WANG,Masako MATSUMOTO,Maki NAGATA,Kenshin HIROMATSU,Kuniyoshi SHIMIZU,
Isolation, Characterization and in Vitro Antioxidant Activity of Polysaccharide from Pholiota adiposa, 第70回日本木材学会, 2020.03, Pholiota adiposa is a species of edible fungus in the family Strophariaceae. It is a valuable mushroom that is grown only in Fukuoka. Polysaccharides extracted from P. adiposa were reported to have immune-stimulatory and antitumor properties, as well as hypolipidemic effect, anti-oxidant and anti-aging activities. We decide to conduct a more systematically studies of P. adiposa Polysaccharide (PAP), including the extraction, isolation, purification, characterization and in vitro antioxidant activity of purified polysaccharide. Crude polysaccharide of P. adiposa was extracted by hot water extraction and alcohol precipitation, and deproteinized by Sevag method. The crude polysaccharide of P. adiposa was further purified using Diethylaminoethanol (DEAE) Sepharose FF and Sephacryl S-200 HR column chromatography. After UV-visible spectrophotometry and gel permeation chromatography (GPC) analysis, the separated polysaccharide was determined to be a purified polysaccharide of 95% and 99% purity. The molecular weight was determined using pullulan standards (molecular weight T-736, 348, 200, 113, 48.8, 23.0, 9.9, 6.6 KD) and the monosaccharide composition was analyzed by HPLC-ELSD. The isolates were analyzed by Fourier Transform Infrared Spectroscopy (FT-IR), Nuclear Magnetic Resonance(NMR) for structural analysis. Then, the antioxidant ability of the polysaccharide fractions was evaluated by the ORAC method and the DPPH method. .
4. YHIYA Amen,AHMED Ashour,ROGERS Mwakalukwa,NAOMICHI Takemoto,MASUMI Nagae,KENSHIN Hiromatsu,KOICHIRO Ohnuki,KUNIYOSHI Shimizu, Anti-Hyperglycemic Effect of Pholiota adiposa on Human Subjects and isolation of active components, 第70回日本木材学会, 2020.03, Mushrooms are known to possess bioactive compounds responsible for anti-hyperglycemic effect. An in vitro α-glucosidase inhibition assay of 8 different mushroom species, resulted in the recognition of the methanolic extract of Pholiota adiposa as the strongest inhibitor. A randomized placebo controlled clinical trials performed in 15 healthy Japanese adults, showed significant dose dependent decrease in the blood glucose level, after consumption of 25 g and 50 g fresh equivalent of P. adiposa. Deep investigation of the methanolic extract of P. adipose, guided by an in vitro α-glucosidase inhibitory assay was performed for isolation of the active components. The n-hexane-soluble fraction of the methanolic extract was found active and lipids-like constituents of this fraction were found to be responsible for the activity of the extract. These results will be valuable in establishing P. adiposa as an important source of bioactive compounds capable of suppressing postprandial hyperglycemia in patients with Type 2 diabetes..
5. MWAKALUKWA Rogers,AMEN Yhiya,SHIMIZU Kuniyoshi, Postprandial hyperglycemia lowering effect of metabolites isolated from olive mill wastes (OMW)―An inhibitory activity on α-glucosidase and α-amylase enzymes―, 第70回日本木材学会, 2020.03, Diabetes is characterized by chronic hyperglycemia resulting from either, when the pancreas does not produce enough insulin (type 1), or when the body cannot effectively use the insulin it produces (type 2). The raise of blood glucose level, postprandially, results from hydrolysis of carbohydrates catalyzed by α-glucosidase and α-amylase. Thus, inhibition of these enzymes results into massive reduction of postprandial blood glucose levels ? a major cause of diabetes. In the present study, we report the isolation and anti-diabetic activity (inhibitory effect on α-glucosidase and α-lipase) of eight (8) compounds, one of them being novel, from olive mill wastes (OMW). Additionally, we investigated the enzyme kinetic analysis of the most active compounds to assist the possible characterization of the inhibition mechanisms involved. We observed that four compounds, 1, 2, 4 and 7, had higher activity against both enzymes. The kinetics studies of these compounds revealed that oleanolic acid 1 and luteolin-7-O-glucoside 7 inhibit α-glucosidase in a mixed-type fashion, while 1-acetoxypinoresinol 4 does it uncompetitively. Both mixed type and uncompetitive inhibition are similar to noncompetitive inhibition except that binding of the substrate or the inhibitor affect the enzyme’s binding affinity for the other. Thus for both, mixed type and uncompetitive, Ki value, which measures the affinity of the inhibitor for an enzyme was greater than 1 (Ki >1). Even though the novel compound, 3,4-dihydroxyphenyl-2-ethoxyethanol 5 exhibited poor activity, it helped to study the SAR for compounds of same class (hydroxytyrosol acetate 6 and hydroxytyrosol), whereby we clearly observed that both methoxylation and esterification of different positions does not influence their activity against the enzymes. This work attracts more attention to OMW as a potential source of lead compounds for structural modification to improve and develop new anti-diabetic drugs..
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