N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry

2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology”

Book of Abstracts

March 15-18, 2010

Novosibirsk, Russia

N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, Russia Korea Institute of Science and Technology, Gangneung, Republic of Korea Gangneung Science Industry Foundation, Republic of Korea Novosibirsk State University, Russia

2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology”

Novosibirsk, Russia March 15-18, 2010

Book of Abstracts

Novosibirsk, 2010

Abstracts of Papers of the 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology”

The most of abstracts are printed as presented, and all responsibilities we address to the authors. Some abstracts underwent a correction of misprints and rather mild editing procedure.

Edited by Prof. Elvira E. Shultz and Dr. Aleksandr V. Shpatov

Compiled by Elena P. Romanenko

Copyright © 2010 N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry

Organizing Committee

Chairman: Prof., Dr. Igor A. Grigoriev Novosibirsk Institute of Organic Chemistry, Novosibirsk, Russia Co-Chairmans: Dr. Byung Hun Um Korea Institute of Science and Technology, Gangneung, Republic of Korea

Dr. Sergey A. Popov Novosibirsk Institute of Organic Chemistry, Novosibirsk, Russia Scientific Secretary: Dr. Konstantin P. Volcho Novosibirsk Institute of Organic Chemistry, Novosibirsk, Russia Members: Prof. Vladimir A. Reznikov (Novosibirsk State University, Russia) Prof. Elvira E. Shults (Novosibirsk Institute of Organic Chemistry, Russia) Prof. Alexey Ya. Tikhonov (Novosibirsk Institute of Organic Chemistry, Russia) Prof. Alexey V. Tkachev (Novosibirsk Institute of Organic Chemistry, Russia) Dr. Irina L. Anisimova (Novosibirsk Institute of Organic Chemistry, Russia) Dr. Irina V. Kolesnikova (SiNaM Ltd., Novosibirsk, Russia) Dr. Mikhail M. Mitasov (Novosibirsk Institute of Organic Chemistry, Russia) Dr. Natalya A. Orlova (Novosibirsk Institute of Organic Chemistry, Russia) Dr. Artem D. Rogachev (Novosibirsk Institute of Organic Chemistry, Russia) Dr. Alexander V. Shpatov (Novosibirsk Institute of Organic Chemistry, Russia) Dr. Evgeny V. Suslov (Novosibirsk Institute of Organic Chemistry, Russia) Dr. Olga I. Yarovaya (Novosibirsk Institute of Organic Chemistry, Russia) Elena P. Romanenko (Novosibirsk Institute of Organic Chemistry, Russia)

International Scientific Board Prof., Academ. Valentin A. Stonik (Pacific Institute of Bioorganic Chemistry, Vladivostok, Russia) Prof., Academ. Genrikh A. Tolstikov (Novosibirsk Institute of Organic Chemistry, Russia) Prof., Academ. Boris A. Trofimov (Irkutsk Institute of Chemistry, Russia) Academ. NAS RK Sergazy M. Adekenov (Research and Production Center "Phytochemistry" , Karaganda, Kazakhstan) Dr. Byung Hun Um (Korea Institute of Science and Technology, Gangneung, Republic of Korea) Prof. Igor A. Grigoriev (Novosibirsk Institute of Organic Chemistry, Russia) Prof. Shomansur Sh. Sagdullaev (Institute of Chemistry of Plant Substances, Tashkent, Uzbekistan) Prof. Elvira E. Shults (Novosibirsk Institute of Organic Chemistry, Russia) Prof. Alexey V. Tkachev (Novosibirsk Institute of Organic Chemistry, Russia)

Conference Secretariat: N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry Acad. Lavrentjev Avenue 9, Novosibirsk 630090, Russia Tel: +7(383)3308870 (9:00-17:00, Time Zone - GMT +06:00) Fax: +7(383)3309752 (twenty-four-hour) E-mail: [email protected], [email protected] URL: http://www.nioch.nsc.ru/conf2010/

The organizers express their gratitude to

Russian Foundation for Basic Research Gangneung Science Park, Republic of Korea

Sinam Ltd., Novosibirsk, Russia

Donau Lab., Zurich, Switzerland

Promix Ltd., Novosibirsk, Russia

MACC center, Republic of Korea Squid industry center, Republic of Korea Lottee wine company, Republic of Korea “Vedun’ya” Ltd., Novosibirsk, Russia for the support of the Conferences

Content

Content

Plenary Lectures New Sesquiterpene Lactones of Plants. Structure of Molecules, Properties, Distribution in the Nature Sergazy M. Adekenov...... 1 Tree Raw Material from Siberia as a Source of Immune System, Growth and Development Stimulators for Use on Plants N.A. Kolchanov, S.E. Peltek, I.V. Mikhailapov, V.E. Kozlov, V.S. Koval, A.V. Kochetov, E.I. Chernyak, S.V. Morozov, M.M. Mitasov, I.A. Grigor’ev...... 3 Successes in Chemistry of Tricyclic Quinazoline Alkaloids and Their Analogues Kh.M. Shakhidoyatov, B.Zh. Elmuradov, A.Sh. Abdurazakov, Kh.A. Bozorov, Zh.E. Turdibaev, A.O. Nasrullaev...... 4 LC-NMR-MS on Natural Products Chemistry Research – Possibility and Limits Byung-Hun Um, Suk-Woo Kang, Sang-Min Kim ...... 5

Oral Communications The Bioconversion of Native Starches Vladimir V. Aksyonov, Anastasia V. Maximenko...... 7 Chemical Constituents of Hammada elegans (Bunge) Botsch and Their Antimicrobial Activity Nabila Abduaziz Al-Jaber ...... 8 Synthesis and Biological Activity of New Derivatives of Mono- and Sesquiterpenoides of Essential Oils Gayane A. Atazhanova ...... 9 Phytochemical Study of Cyperus alternifolius L. as a hepatoprotective Plant Against Carbon Tetrachloride-Induced Hepatotoxicity in Rats Amani S. Awaad...... 10 Flavonoids and Polysaccharides of Larch Wood, Isolation and Purification Technology and Experimental-Industrial Production Creation V.A. Babkin ...... 11 Anticoagulative Effect of the Preparation of Serine Proteases Inhibitor from Hepatopancreas of King Crab (Paralithodes camchaticus) Maria V. Balashova, Galina N. Rudenskaia, Ylia A. Rudenskaia, Viacheslav A. Isaev, Lydmila V. Lytova ...... 12 Skeletal and Oxidative Transformations of Some Lead Triterpenic Acids for the Structure-Anticancer Activity Relationships Studies M.V.Khudobko, L.R.Mikhailova, L.A.Baltina ...... 13 Seasonal Fluctuations of Phycobiliproteins and Sulfated Polysaccharides in Ahnfeltiopsis flabelliformis (Harv.) Masuda. (Rhodophyta, Phyllophoraceae) Anna O. Barabanova, Anna O. Kravchenko, Irina M. Yakovleva, Irina M. Yermak ...... 14 St. John’s Wort Flowers Composition Irina I. Bayandina, Tatyana P. Kukina ...... 15 Cellulose from Cereal Waste and Miscanthus Biomass Vera V. Budaeva ...... 16 Biosynthesis of Astaxanthin in Heterologous Hosts by Marine Bacterial crtW and crtZ Genes Seon-Kang Choi and Norihiko Misawa...... 17 Optimization of Gelatin-Gum Arabic Complexed Microcapsules for Encapsulation of Allyl Isothiocyanate Zhi-Qiang Zhang, Cheol-Ho Pan, and Donghwa Chung...... 18 Anticarcinogenic Activity of Aaptamine and Its Derivatives Isolated from Marine Sponge Aaptos sp. Content

Sergey A. Dyshlovoy, Sergey N. Fedorov, Larisa K. Shubina, Valentin A. Stonik ...... 19 Sulphated Polysaccharides of Brown Seaweeds. History and Modern Day of the Investigation Svetlana P. Ermakova...... 20 New “Green” Way to Epoxydes of Terpenes Vladislav V. Fomenko, Oleg V. Bakhvalov, Sergey S. Laev, Nariman F. Salakhutdinov...... 21 Plant Coumarins of Some Apeaceae Species Growing in Siberia and Mongolia J. Ganbaatar, E.E. Shults, D. Mungunnaran, A. Tuvshintugs, G.A. Tolstikov...... 22 Study on Chemical Constitutes of the Volatiles from Dracocephalum nutans L. Jenis Janar, G. Sh Burasheva, Haji Akber Aisa ...... 23 Methodology of Cytostatic Drug Development on the Basis of Chromatographic Isolation and Hightroughput Screening of Plants Extracts S.G. Klochkov...... 24 A Developed Technology of Fir Needles Complex Processing Tatyana P. Kukina, Tamara A. Vaganova, Eugeny V. Malykhin, Sergey A. Popov, Andrey M. Chibiryaev ...... 25 Mechanochemistry of Plant Materials Oleg I. Lomovsky...... 26 Some Studying Results of Kazakhstan Wild-Growing Plants and the Ways of Their Research D.U. Korulkin, G.A. Abilov, R.A. Muzichkina...... 27 Contrastive Analysis of Composition and Bactericidal Activity of Various Fraction of Siberian Pine Essential Oil Anton N. Narchuganov, Anna A. Gontova, Alexander A. Efremov...... 28 Doping agents and Oriental Herbs in Korea Jae Keun Oh...... 29 Synthesis and Cytotoxic Activity of 7,10-Dihydroxy-12H-benzo[b]xanthene-6,11-dione Derivatives Analogs of Bikaverin Dmitry N. Pelageev, Alexandra S. Kuzmich, Victor Ph. Anufriev ...... 30 Gene-Design for the Optimal Expression in Yeast of the Genetically Modified Constructions Including preS and S-genes of A and D Genotypes of Hepatitis B Virus Svetlana V. Polyarush, Shakhnoz S. Azimova...... 31 Content of Alkaloids in the Different Parts of Sophora flavescens Soland Denis V. Sandanov, Natalia A. Pankrushina ...... 32 Synthetic Transformations of Alkaloids, Terpenoids and Coumarins of Siberian Flora for Search of New Pharmaceutics Elvira E. Shults...... 33 A New Multifunctional Agents Synthesized on Natural Triterpenoid Platforms I.V. Sorokina, T.G. Tolstikova, D.S. Baev, G.A. Tolstikov...... 34 The Study of Antioxidant Activity of Natural Organic Acids O.N. Tchaikovskaya, I.V. Sokolova, L.V. Nechaev, N.V. Yudina, E.V. Mal’tseva, E.A. Sosnin, S.M. Avdeev ...... 35 Medications Standardization Problems N.A. Vereshchagin ...... 36 New in Ecdysteroids Chemistry: the Synthesis of Minor Phytoecdysteroids and Ecdysteroids Analogues Natalya A. Veskina, Aliya Sh. Ibragimova, Ilgiz V. Galyautdinov, Victor N. Odinokov...... 38 Resources and Biotechnology of the Ecdysteroid-Containing Plants Svetlana O. Volodina, Ivan F. Chadin, Vladimir V. Volodin ...... 39 Biological Activity of Sulfated Polysaccharides from Red Seaweed of Pacific Coast Irina M. Yermak, Anna O. Barabanova, Victoria N. Davidova, Ekaterina V. Sokolova, Yong Hwan Kim, Kwang Soon Shin...... 40 Content

Synthesis of Biological Active [N-(2-Ethoxyethyl)piperidyl-4]propargyl Derivatives of Natural Alkaloids and Their Synthetic Analogs Valentina K. Yu, Kaldybay D. Praliyev, Aissulu Zh. Kabdraissova, Kanitar Kanitar...... 41 Isolation of Substances from Plants of Hipophae rhamnoides L. Species and Study on Their Phytochemical and Antioxidant Properties Gaukhar S. Sapargalieva, Galiya E. Zhusupova, Maira K. Murzakhmetova, Zharylkasin A. Abilov, Zulkhair A. Mansurov ...... 42

Poster Presentations Synthesis and Biological Activity of Sesquiterpene Lactone Modified by Natural Alkaloids S.G. Klochkov, S.V. Afanasyeva, A.B. Ermatova ...... 43 Chemical Constituents of Leonurus turkestanicus I.M. Isaev, M.A. Agzamova and M.I. Isaev...... 44 Anticancerogenic Activity of Alkaloids from Haplophyllum perforatum V.I. Akhmedjanova, L. Angenot, R.Sh. Shakirov...... 45 Quantification of Ecdysteroids in Insect by HPLC-MS Alexander A. Alekseev, Vladimir G. Vasiljev...... 46 Chemical Composition of the Essential Oil of Thymus punctulosus L.I. Alekseeva, A.G. Bystrushkin, I.V. Gruzdev...... 47 Improving Effects of Semen Flax Extract on Hemorheological Abnormalities in Ovariectomized Rats Anna M. Anischenko, Tatyana M. Plotnikova, Natalia V. Vasil’eva ...... 48 Synthesis of C-20 Pyridine Analogs of Lupane-Type Triterpenoids Anna N. Antimonova, Natalya I. Petrenko, Elvira E. Shults, Genrikh A. Tolstikov ...... 49 Synthesis of Six 3-Methyl-6-(1-methylethenyl)cyclohex-3-en-1,2-diol Stereoisomers with High Optical Purity O.V. Ardashov, I.V. Il’ina, K.P. Volcho, N.F. Salakhutdinov...... 50 Gas-Chromatographic Analysis of the Volatile Compounds of Halostachys belangeriana G.Zh. Baisalova, G. Ozek, K.H.C. Baser, S.B. Rakhmadieva...... 51 Hypericum perforatum L. as Potential Antidiabetic Medicinal Herb Irina I. Bayandina, Tatyana P. Kukina ...... 52 The Synthesis of Hybrid Structures Consisted of Sesquiterpene Methylene Lactone and Alkaloid Alexander V. Belovodskii, Elvira E. Shults, Khusnutdin M. Shakhidoyatov, Genrih A. Tolstikov ...... 53 On Phytoadaptive Activity of Lonicera caerulea L. Irina G. Boyarskikh and Larisa A. Yevtukhova...... 54 Enzymatic Hydrolysis of Miscanthus Conversion Products Vera V. Budaeva ...... 55 Synthesis and Antioxidant Activity of Alkylthiomethyl Derivates of Quercetin and Dihydroquercetin Ivan M. Bugaev, Aleksandr E. Prosenko...... 56 Obtaining Mannanooligosaccharide Additive by Means of the Mechanoenzymatic Hydrolysis of Yeast Biomass A.L. Bychkov, E.I. Ryabchikova, K.G. Korolev, O.I. Lomovsky...... 57 Research of Siberian Fir Phenol Compounds Using Chromatographic Profiling Method E.I. Chernyak, A.I. Vyalkov, S.V. Morozov...... 58 Steam-distillation and Simultaneous Hydrodistillation – Solvent Extraction in Essential Oil Preparation from Fresh Plant Material Dmitry V. Domrachev, Alfred L. Revutsky, Alexey V. Tkachev ...... 59 Enantiomeric Composition of Monoterpene Hydrocarbons Isolated from Siberian Conifers Dmitry V. Domrachev, Elena V. Karpova, Alexey V. Tkachev ...... 60 Content

Chromatographic Isolation and Hightroughput Screening of Potencial Cytostatic on the Base of Extracts from Asteraceae Plants A.B. Ermatova, A.V. Chudinov, O.V. Novikova, I.M. Bravova, S.G. Klochkov...... 61 Clerodane Diterpenoids from Pulicaria Gnaphalodes K.A. Eshbakova...... 62 Flavonoid from Scutellaria holosericea H.Sh. Kamoldinov, K.A. Eshbakova ...... 63 Qualitative Analysis of Coumarins in Seeds Nigella sativa L., Growing in Uzbekistan K.A. Eshbakova, Sh.A. Temurova, I.K. Azizov, N.D. Abdullaev...... 64 Flavonoid from Roots Scutellaria schachristanica Z.O. Tashmatov, K.A. Eshbakova, Kh.M. Bobakulov, N.D. Abdullaev...... 65 Optimization of Method of Isolation of Leucomisine from Artemisia leucodes Schrenk Vladimir V. Folmer, Ilya A. Khabarov, Bibigul B. Rakhimova, Svetlana A. Ivasenko, Anar N. Zhabaeva, Khorlan I. Itzhanova, Sergazy M. Adekenov...... 66 Preparation of -Pinene Oxide from Turpentine Sergey S. Laev, Vladislav V. Fomenko, Nariman F. Salakhutdinov...... 67 Chemical Research of Marsh Cinquefoil (Comarum palustre) Grows in the Krasnoyarsk Territory Sergey V. Golubev, Alexander A. Efremov ...... 68 Ultrasound Assisted Enzymatic Hydrolysis of Cellulose as a Method of Improvement of Biologically Active Substances Extraction O.V. Golyazimova, A.A. Politov, O.I. Lomovsky ...... 69 Mannich-Type Three Component Condensation of α-Substituted Caran-4-one Oxime with Formaldehyde and Secondary Amine Nikolay B. Gorshkov, Alexandr M. Agafoncev...... 70 Quantitative Determination of Betulin and Lupeol in Birch Bark Extracts by HPLC Alexander E. Grazhdannikov, Ekaterina A. Khokhrina, Olga P. Sheremet ...... 71 Unusual Reactions of (+)-2- and (+)-3-Carenes with Aldehydes on K10 clay Irina V. Il’ina, Konstantin P. Volcho, Dina V. Korchagina, Georgi E. Salnikov, Alexander M. Genaev, Elena V. Karpova and Nariman F. Salakhutdinov...... 72 Chemical Transformation of Cycloartane Triterpenoids I.M. Isaev, D.A. Iskenderov, M.A. Agzamova and M.I. Isaev...... 73 Perspectives of Ecdysten Use in the Treatment of Giardiasis as Mono- and Co-infection Jannat I. Islamova, Vladimir N. Syrov, Zainab A. Khushbaktova, Svetlana O. Osipova...... 74 Investigation of Capillary-Protective Composition on the Base of Dihydroquercetin and Lipoic Acid Ivan S. Ivanov, Anna M. Anishenko, AnastasiaV. Sidekhmenova, Mark B. Plotnikov...... 75 Investigation of Biological Activity of the Stereoisomeric Halogenhydrines of Ludartine Roza I. Jalmahkanbetova ...... 76 Synthesis of Tetrahydroquinoline-Barbituric Acid-Terpene Hybrids Aliaksei P. Kadutskii and Nikolas G. Kozlov...... 77 One-pot Synthesis of 3-Aminoacridine Derivatives Aliaksei P. Kadutskii and Nikolas G. Kozlov...... Ошибка! Закладка не определена. Obtain and Properties of Cryogel of Polyvinyl Alcohol A.A. Kasymbekov, M.T. Omyrzakov, B.M. Kudaibergenova, Sh.N. Zhumagalieva, M.K. Beisebekov, Z.A. Abilov...... 79

Isolation of Arglabin from CO2-Extraction of Artemisia glabella Kar. et Kir. by Use of Flash- Chromatography Ilya A. Khabarov, Svetlana A. Ivasenko, Bibigul B. Rakhimova, Sergazy M. Adekenov...... 80 Ferula foetida (Bunge) Regel Growing in Uzbekistan Content

E.Kh. Khalilova, O.Sh.Yusupov, B.T. Sagdullaev, Kh.S. Akhmedkhodjaeva, N.Sh. Davlatov, S.F. Aripova...... 81 Synthetic Modification on the B Ring of Labdane-Type Diterpenoids Yurii V. Kharitonov, Elvira E. Shults, Genrikh A. Tolstikov ...... 82 Studying Alkaloids of Haplophyllum griffithianum D.R. Kodirova, H.A. Rasulova, R. Shakirov ...... 83 Chemical Fixation of Betulonic Acid on Water Soluble Polyvinyl Alcohol Tamara I. Kogay, Tatiana L. Sostina, Boris N. Kuznetsov...... 84 Synthesis of New Esters of Peoniflorin, the Major Monoterpene Glycoside of Paeonia anomala L. Roots R.M. Kondratenko, Z.G. Kulmukhametova, O.V. Stolyarova, L.A. Baltina (jr.), L.A. Baltina ...... 85 Synthesis of New Chiral Ligand from (+)-2-Carene Ekaterina A. Koneva, Konstantin P. Volcho, Yuri V. Gatilov, Dina V. Korchagina, Nariman F. Salakhutdinov ...... 86 The New Method of Salicylates Extraction Anna V. Koptina, Alexey I. Shurgin, Albert V. Kanarskiy...... 87 Triterpenoids of Picea and Pinus Species Tatyana P.Kukina, Emma N. Shmidt ...... 88 Synthesis of Betulin Esters by Interaction of Organic Acids with Birch Bark Svetlana A. Kuznetsova, Galina P. Skvortsova, Boris N. Kuznetsov...... 89 New Methods of Allobetulin Synthesis Vladimir A. Levdansky, Svetlana A. Kuznetsova, Galina P. Skvortsova, Alexander V. Levdansky, Boris N. Kuznetsov ...... 90 Palladium-Catalyzed Amination in the Synthesis of Diamino Furocoumarin Derivatives Alla V. Lipeeva, Elvira E. Shults, Genrikh A. Tolstikov...... 91 The Flavonoids of Some Species of Kazakhstan Rumex L. Y.A. Litvinenko, R.A. Muzychkina...... 92 Solid State Mechanochemical Reaction of Hypericin in Plant Material I.O. Lomovsky, O.I. Lomovsky...... 93 Phytochemical study of Artemisia pontica L. Darya L. Makarova, Minisa A. Khanina, Dmitry V. Domrachev, Alexey V. Tkachev ...... 94 Extractive Compounds of Punica granatum L. Flowers N.M. Mamatkulova, F.A. Melikuziev, S.Z. Nishanbayev, N.K. Kh idirova, S.F. Aripova, Kh.M. Shakhidoyatov...... 95 Dissolution Rate Control of Nitrogen Containing Forage Additives Serguey G. Mamylov, Oleg I. Lomovsky, Victor A. Rogachev, Vladimir A. Soloshenko...... 96 Steroid Glycosides in Some Allium L. Species of Alliaceous Family (Alliaceae J. Agardh) N.V. Matistov, T.I. Shirshova, I.V. Beshley...... 97 Sterols as a Part of Neutral Lipids in Some Allium L. (Alliaceae) Species N.V. Matistov, T.I. Shirshova, I.V. Beshley...... 98 Substance from Plant Limonium Myrianthum as the Basis for Creation of New Medical Products Kseniya Mayeva, Anastassiya Gadetskaya, Zhanar A. Kozhamkulova, Galiya E. Zhusupova, Zharylkasin A. Abilov...... 99 Preparation and Solid-State Characterization of Solvates of Betuline and Its Mechanocomposites with Polymers M.A. Mikhailenko, T.P. Shakhtshneider, V.A. Drebushchak, T.N. Drebushchak, S.A. Kuznetsova, V.V. Boldyrev ...... 100 Reaction of (-)-Cytisine with Thiourea A.V. Minilbaeva, A.N. Lobov, I.P. Tsypysheva, M.S. Yunusov ...... 101 Naphthoquinone Substituted Labdanes via Diels-Alder Reaction Content

Maksim E. Mironov, Elvira E. Shults, Yurii V. Kharitonov, Genrikh A. Tolstikov...... 102 Alkaloids of Korolkowia Sewertzowi – Original Antiarrhythmic Drugs of Different Mechanisms of Action Yu. R. Mirzaev, R. Shakirov, D.U. Abdullaeva...... 103 The Study of Qualitative and Quantitative Characteristics of the Kazakhstan Type of Plant, Polygonum Alpinum A.E. Mussina, R.A. Muzichkina...... 104 Correction of Hypoxia of States at Animals Individual and Integral Materials from the Plant Convolvulus subhirsutus A.N. Nabiev, S.F. Aripova, A.M. Gapparov, V.U. Khujaev, M.U. Alikulova...... 105 Obtaining and Hepatoprotective Action of the Drug from Codonopsis clematideae A.N. Nabiev, B.T. Sagdullaev, S.F. Aripova, Z.A. Hushbaktova, V.N. Syrov ...... 106 Influence of Steroidal Alkaloids on the Steroidogenesis at Rats A.N. Nabiev, R.Sh. Shakirov, D.U. Abdullaeva...... 107 Synthesis of Bromo- and Iodo-Berberine Ivan V. Nechepurenko, Nina I. Komarova, Marina P. Polovinka, Nariman F. Salakhutdinov, Genrikh A. Tolstikov...... 108 Synthesis and Antioxidant Activity of Alkylthiomethyl Derivates of Hydroquinone, Resorcinol and Pyrocatechol Sergey L. Nesterovich, Elena V. Korobicina, Aleksandr E. Prosenko ...... 109 A Convenient Synthesis of 3- and 3,10-Functionalized Indolizino[8,7-b]indole Derivatives from Harmine Z.S. Nurmaganbetov, E.E. Shults, S.V. Chernov, A.Z. Turmukhambetov, A.V. Kazantsev, G.A. Tolstikov, S.M. Adekenov...... 110 Synthesis of 3-Hydroxy-4-[hydroxy(phenyl)methyl]-3-methyl-3,4-dihydrofuro[3,2-c]coumarin Irina A. Os’kina, Yurij V. Gatilov, Alexsei Ya. Tikhonov ...... 111 Alkaloids and Flavonoids from Roots of Sophora flavescens Soland. Isolation and Identification Natalia A. Pankrushina, Vladimir G. Vasiljev, Elena I. Chernyak, Denis V. Sandanov, Vladimir V. Boldyrev, Byung Hun Um ...... 112 Novel Methods of Mechano-Enzymatic Modification and Recovery of Agriculture Waste A.A. Politov, O.V. Golyazimova ...... 113 Unique Siberian Soybean Cultivar SibNIIK-315 and Tendency of Plant Breeding Nikolay I. Kashevarov, Revmira I. Polyudina, Dmitry A. Potapov, Olga A. Rozhanskaya...... 114 Effects of Hybrid Macromolecular Phenolic Antioxidants on Whole Blood Viscosity Elena V. Popova, Mark B. Plotnikov, Oleg I. Aliev, Nina S. Domnina, Olga V. Sergeeva...... 115 Ecdysteroids of Serratula quinquefolia Bieb. ex Willd Yana I. Pylina, Svetlana O. Volodina, Dmitry S. Bacharov, Vladimir V. Volodin ...... 116 Working out Optimal Technology for the Substance Isolation from Cuscuta campestris Yuncker Aizhan I. Zhusupova, Saniya N. Rakhadilova ...... 117 Synthesis of N-(12-Cytisinylethyl)-1-(4-methoxyphenyl)-6,7-dimethoxy-1,2,3,4- tetrahydroisoquinoline and Computer Prediction of Its Biological Activity (PASS) Sh.B. Rakhimov, V.I. Vinogradova ...... 118 Chemical Study of Artemisia succulenta Ldb. Bibigul B. Rakhimova, Zhanar R. Shaimerdenova, Sergazy M. Adekenov...... 119 Antimicrobic Activity in vitro of the Extracts Containing Phytoecdysteroids I.D. Bobaev, A.M.-T. Bektimirov, I.M. Boymatov, N.Sh. Ramazanov ...... 120 Two Major Stilbene Glycosides of Pinus sibirica Bark: Isolation and Chemical Transformation Alexandra V. Pozdeeva, Nina I. Komarova, Artem D. Rogachev, Dina V. Korchagina, Nariman F. Salakhutdinov, and Genrikh A. Tolstikov ...... 121 Content

Distribution of Enantiomers of Monoterpene Hydrocarbons in Siberian Plants of the Apiaceae (Umbelliferae) Family Elena P. Romanenko, Elena V. Karpova, Elena A. Korolyuk, Alexey V. Tkachev ...... 122 Synthesis of New Lappaconitine Derivatives Containing Indolizine Framework Vasiliy E. Romanov, Elvira E. Shults, Genrikh A. Tolstikov ...... 123 Testing Cytokinin Activity of Rice Husk Mechanochemical Preparation with the Help of Culture Chickpea (Cicer arietinum L.) in vitro Olga A. Rozhanskaya, Oleg I. Lomovsky ...... 124 Cathepsin L–Like Protease from Dermestid Beetle Dermestes frischii Maggot G.N. Rudenskaya, A.S. Javadov, Yu. A. Rudenskaya, A.I. Papisova, G.N. Balandina, R.D. Zhantiev, O.S. Korsunovskaia, Y. E. Dunaevsky ...... 125 Synthesis and Spatial Structure of 4-(2-Isothiocyanatopropan-2-yl)methyl-1-thiocyanatocyclohexane Daniyar T. Sadyrbekov, Oleg G. Ryazantsev, Yelena V. Tikhonova, Almagul Mahmutova, Koblandy M. Turdybekov, Yurii V. Gatilov, Gayane A. Atazhanova, Sergazy M. Adekenov ...... 126 Dependence of N-Desacetyllappaconitine Hydrobromide from the Method of Its Preparation Sh.Sh. Sagdullaev, A.Z. Sadikov, N.V. Valiev...... 127 Analytical Control of Production of N-Desacetyllappaconitine Hydrobromide A.Z. Sadikov, Sh.Sh. Sagdullaev, N.V. Valiev, Sh.A. Otaeva...... 128 Chromatographic Method of Purifying Substance of Acsaritmin A.Z. Sadikov, Sh.Sh. Sagdullaev, Z.U. Mullabaeva...... 129 Spectrophotometric Method of Analysis of Substance of Acsaritmin Preparation A.Z. Sadikov, Sh.Sh. Sagdullaev, Z.U. Mullabaeva, Sh.A. Otaeva...... 130 BAS Accumulation and in vitro Propagation of Salix High Productive Genotypes Roman V. Sergeyev, Nadezhda N. Sheveleva, Aleksey I. Shurgin...... 131 Distribution of Saponins in Fabaceae Dmitry M. Shadrin, Alexander V. Rodionov, Svetlana O. Volodina, Kirill G.Tkachenko, Vladimir V. Volodin..... 132 Polyprenols of Cotton Leaves and Their Pharmacological Activity Kh.M. Shakhidoyatov, N.K. Khidyrova, N.M. Mamatkulova, V.N. Syrov, Z.A. Khushbaktova...... 133 Isolation of Betulin from Birch Bark by Supercritical Alcohols Viktor I. Sharypov, Sergey V. Baryshnikov, Natalia G. Beregovtsova, Boris N. Kuznetsov...... 134 Cryoconservation of Cultures of Cells of Endemic Species of Plants Z.K. Shaushekov, A.Zh. Kuandykova, A.B. Eshmagambetova, G.K. Asanova, S.M. Adekenov ...... 135 Step-by-Step Preparation of Spironorketone from 2,3-Seco-19β,28-epoxy-28-oxo-18α-olean-2,3- dicarboxylic anhydride Andrew V. Shernyukov, Iliya Ya. Mainagashev, Dina V. Korchagina, Nariman F. Salakhutdinov, Genrikh A. Tolstikov...... 136 Extractive Substances from Twigs of Picea obovata L. Emma N. Shmidt, Galina F. Chernenko...... 137 Influence of Oxime Pinostrobine on Lipidic Spectrum at Rats with Experimental Diabetes Leila I.Arystan, Zarina T. Shulgau...... 138 The Neutral Compounds of Aerial Parts of Geranium saxatile Kar et Kir. D.R. Siddikov, N.M. Mamatkulova, S.Z. Nishanbayev, N.K. Khidirova, Kh.M. Shakhidoyatov ...... 139 Microcalorimetric Method for Study of Antioxidant Activity of Natural Lipid Fractions Natalia V. Sizova...... 140 Oxidation Usnic Acid by Organic Peracids Dmitry N. Sokolov, Olga A. Luzina, Marina P. Polovinka, Nariman F. Salakhutdinov...... 141 Chemical Contaminations of Camhorosma monspeliacum A.K.Umbetova, N.A. Sultanova, G.Sh.Burasheva, Zh.A.Abilov ...... 142 Content

The Separation of Biological Active Complexe from Genus Tamarix by Using Nanostructure Sorbents N.A. Sultanova, G.E. Zhusupova, Zh.A. Abilov, Zh.A. Mansurov, M.A. Beisenbaev...... 143 The Basidiomycete Lentinula edodes Lectins Interaction with the Selected Organoselenium Diphenyl Substances Alexei N. Pankratov, Olga M. Tsivileva, Boris I. Drevko, Valentina E. Nikitina ...... 144 A Preparative Method for Isolation of Fucoxanthin from the East Sea Brown Algae Eisenia bicyclis by Centrifugal Partition Chromatography Sang Min Kim, Ya Fang Shang, Byung-Hun Um...... 145 Preparative Separation of Chlorogenic Acid by Centrifugal Partition Chromatography from Highbush Blueberry (Vaccinium corymbosum L.) Sang Min Kim, Ya Fang Shang, Byung-Hun Um...... 146 Anthocyanins from Azalea (Rhododendron mucronulatum) Flowers Je Seung Jeon, Byung-Hun Um...... 147 The rapid identification of anthocyanins from blueberry, Vaccinium corymbosum by LC–NMR/MS Suk Woo Kang, Sang Min Kim, Byung-Hun Um...... 148 To the Question of Standardization of Anticancer Drug «Leykoefdin» on the Basis of the Herb of Ephedra Equisetina Yevgeniya S. Ushakova ...... 149 Investigation of Standardization of Rosa corymbifera Phytopreparation N.A. Vereshchagin ...... 150 Perspectives of Production of Supplement with Antitumor Effect, Based on Polysaccharides from Brown Algae Olesya S. Vishchuk, Svetlana P. Ermakova, Thinh D. Pham, Bui Minh Ly, Tatyana N. Zvyagintseva...... 152 Biocompozites on the Base of Natural Sorbents, Synthetic and Natural Polymers I.A. Vorsina, T.F. Grigorieva, A.P. Barinova, N.Z. Lyakhov...... 153 Molecular Characteristics of Water-Soluble Sulphated Polysaccharides from Enteromorpha prolifera and Their Anticancer and Immunomodulatory Activities Myeong Lae Cho and Sang Guan You...... 154 Coagulation Ability of Humic Acids from Mechanochemically Activated Peat Alexander A. Ivanov, Natalya V. Yudina, Oleg I. Lomovsky ...... 155 Bismuth Compounds for Medicine Based on Natural Products Yuri M.Yukhin, Olga A. Logutenko, Evgeniya V. Timakova, Ekaterina S. Naydenko, Lubov’ I. Afonina, Tatiana A. Udalova...... 156 Preparation of Monoclonal Antibodies Against the HBsAg from Transgenic B. mori Larvae Elvira G. Yusupova ...... 157 Retina Protection Properties of 4-Methyl-2,6-diisobornylphenol Anna A. Zhdankina, Galina A. Kon, Mark B. Plotnikov, Sergey V. Logvinov, Alexander V. Kuchin, Irena Yu. Chukicheva...... 158 New Flavonol Glycoside from Limonium gmelinii Zhanar A. Kozhamkulova, Mohamed M. Radwan, Galiya E. Zhusupova, Zharilkasin A. Abilov, Samir A. Ross...... 159 Isolation and Identification of 26-Hydroxyecdysteroids Larisa Zibareva, Rene Lafont, Laurence Dinan ...... 160

Author Index…………………………………………………………………..……………….……………………….161

Content

2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Plenary Lecture

Plenary Lectures New Sesquiterpene Lactones of Plants. Structure of Molecules, Properties, Distribution in the Nature

Sergazy M. Adekenov

JSC “International research and production holding “Phytochemistry”, 100009, Karaganda, Kazakhstan Fax: (7212) 43-37-73 E-mail: [email protected]

In territory of Republic of Kazakhstan there are grown over 6000 species of plants from which 667 are endemic as sources of new, before not studied natural compounds. The plants of family Asteraceae Dumort. take a special place in flora of region, where totaled over 1000 species, occupying a significant part of territory and being of interest as potential sources of biologically active sesquiterpene lactones. At JSC “International research and production holding “Phytochemistry” over 80 species of plants of flora of Kazakhstan are studied on the maintenance of sesquiterpene lactones for last years. As perspective sources of sesquiterpene lactones 39 species of plants are determined by results of chemical screening. There are representatives of genera Achillea, Ajania, Artemisia, Centaurea, Chartolepis, Crepis, Gaillardia, Handelia, Inula, Jurinea, Phalacrachena, Pulicaria, Rhaponticum, Saussurea, Stizolophus and Tanacetum of family Asteraceae Dumort. From 26 species of plants it is isolated 67 sesquiterpene lactones, including 11 new lactones: guaianolides artefine (1) from Artemisia filatovii A.Kuprijanov sp. nova, 5β(H)- austricine (2) from Artemisia leucodes Shrenk., raserine (3), raserolide (4), raposerine (5) and 15-deacetylraposerine (6) from Rhaponticum serratuloides (Georgi.) Bobr., germacranolides ajanolide A (7) from Ajania fruticulosa (Ldb.) Poljak. and subchrizine (8) from Artemisia subchrysolepis Filat., pseudoguaianolides gaigranine (9) and gaigrandine (10) from Gaillardia grandiflora Hort., eudesmanolide turneforine (11) from Artemisia tournefortiana Rchb. The structure of molecules of the isolated compounds is determined on the basis of data IR-, UV, 1H-, 13C-NMR – spectroscopy and X-ray structure analysis.

O HO H O O HO OH HO O HO H H HO H O O HO O

O O O 1 2 3

1 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Plenary Lecture

H H OH H OH O O Cl O Cl HO HO HO O O O HO H HO H HO H AcO O AcO O HO O

O O O

4 5 6 OH

AcO AcO O O O O 7 8 O OH O HO H O H

O O

O O O HO OAc HO OAc O 9 10 11

The isolated sesquiterpene lactones are studied on antivirus, cytotoxic, antibacterial, fungicidal, antiparasitic, antimutagenic and phagocytosis-stimulating activities. The perspective samples of compounds of this number with the expressed biological activity are determined for development on their basis the original phytopreparations.

2 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Plenary Lecture Tree Raw Material from Siberia as a Source of Immune System, Growth and Development Stimulators for Use on Plants

N.A. Kolchanov, S.E. Peltek, I.V. Mikhailapov, V.E. Kozlov, V.S. Koval, A.V. Kochetov, E.I. Chernyak, S.V. Morozov, M.M. Mitasov, I.A. Grigor’ev

Institute of Cytology and Genetics, SB RAS, Pr. Lavrentyeva, 10, 630090, Novosibirsk, Russia Novosibirsk Institute of Organic Chemistry, SB RAS, Pr. Lavrentyeva, 9, 630090, Novosibirsk, Russia E-mail: [email protected], [email protected]

Forest resources of Siberia, largely existing as coniferous species, are an immensely rich source of biologically active compounds that have promise for new agents stimulating the immune system, growth and development, which are actively used in agriculture in developed countries for sustainable high yields of cereal and vegetable crops. The use of extraction methods on recoverable plant raw material enhanced by the power of individual and group composition analysis followed by specialized tests allows the coniferous biomass to be effectively utilized. Natural growth, development and immune system stimulators (GDIS) have a number of important advantages: they are biologically safe and have no resource or ecological limitations. The Siberian Branch of the Russian Academy has a long-standing experience in developing new highly efficient growth, development and immune system stimulators from the total processing of the coniferous biomass, which appears as complex mixes of biologically active lipid-phenol-carbohydrate complexes. After many years of research, efficient low-consumption drugs have been developed (Novosil, Larisib, Lariksin, Biofungistim, Abistim, to mention a few), which have demonstrated strong growth- stimulating, fungicidal and stress-reducing activity. The use of these drugs improves crop yield and quality, enhances resistance to adverse factors, ncr i eases labor capacity in agriculture. These drugs activate the molecular-biological and physiological systems that provide for protection against pathogens and adverse effects of abiotic origin. There is a new promising trend in plant developmental genetics called “chemical genomics”, which focuses on the ability of low-molecular-weight compounds to selectively modulate the activity of gene networks controlling the most important biological processes. To be able to address challenges in modern agrobiology, new biotechnological methods are required for the creation of new GDIS, which will be helpful in directing the development of morphological and physiological traits according to a pre-set scenario, in developing mathematical models to describe plant morphogenesis, in identifying critical parameters and, finally, in development new approaches for the programming of plant development up to a pre-set pattern. The data expected to be so obtained will be used towards the most efficient use of new-generation GDIS in agriculture. The production of modern GDIS can be set on the basis of native recoverable plant raw material, which can reduce the manufacturing cost, enhance competitiveness of domestic manufacturers, and improve Russia’s economic and food safety.

3 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Plenary Lecture Successes in Chemistry of Tricyclic Quinazoline Alkaloids and Their Analogues

Kh.M. Shakhidoyatov, B.Zh. Elmuradov, A.Sh. Abdurazakov, Kh.A. Bozorov, Zh.E. Turdibaev, A.O. Nasrullaev

Institute of the Chemistry of Plant Substances, Academy of Sciences, Republic of Uzbekistan, Mirzo Ulugbek str., 77, Tashkent. Fax: 998-71-120-64-75 E-mail: [email protected]

Chemical modification of natural compounds and their synthetic analogues are a perspective direction for creation a new, high performance drugs with various pharmacological activities. Among derivatives of alkaloids of the plants Peganum harmala and Mackinlaya subulata Philipson, and also their thiophene analogues an efficient biologically active compounds are found and introduced into medical practice. In this work a questions on a comparative study of features of reactivity of the tricyclic quinazolines and their thiophene analogues in reactions with electrophilic and nucleophilic reagents proceeding on one of potential reactive centers (a benzene ring, atom of nitrogen N- 1, carbonyl or thiocarbonyl groups at C-4, the methylene groups in -position), revealing of tautomeric forms (imine – enamine, E- and Z-, etc.) of obtained compounds by spectral methods and X-ray analysis will be discussed.

4 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Plenary Lecture LC-NMR-MS on Natural Products Chemistry Research – Possibility and Limits

Byung-Hun Um, Suk-Woo Kang, Sang-Min Kim

Natural Products Research Center, KIST Gangneung Institute, Gangneung 210-340, Korea Fax: 82-33-650-7299 E-mail: [email protected]:

LC-NMR represents an interesting complementary technique to LC-UV-MS for online identification of LC peaks. Many recent applications have demonstrated the usefulness of LC- NMR-MS in the field of natural products chemistry. The recent progress in pulse field gradients and solvent suppression, the improvement in probe technology and the construction of high field magnets have given a new stimulus to LC-NMR, which has emerged since the mid 1990s as a powerful method for the on-line identification of organic molecules. Here, we will present some examples of the application of LC-NMR-MS on Natural product chemistry and discuss its possibilities and limits. The anthocyanins and caffeoylquinic acid are major bioactive compounds in blue berry fruits, and the kinds and contents of anthocyanins are greatly changed in its cultivars and cultivation environment. In common HPLC analysis methods, the standard compounds are absolutely needed for the identification of these anthocyanins. The hyphenated LC-NMR and LC-MS techniques are applied for the direct identification of the major constituents present in buleberry. In search of the anti-inflammatory constituents present in Angelica dahurica extracts, the nitric oxide (NO) inhibitory activity was investigated by HPLC based activity profiling in peak-resolved manner. The hyphenated LC-NMR/MS technique allowed to identify the five known active components from Angelica dahurica extract. Ligularia fischeri (Ledeb.) Turcz., which Korean name is Gomchui, has been used as folk medicines and foods to treat the diseases such as scarlet fever, rheumatoidal arthritis and hepatic function failure in Korea. Gomchi tea was prepared from the leaves of this plant by branching and drying method. The anti-oxidative compounds in Gomchui tea were screened by on-line HPLC-ABTS method and its structures (four caffeoylquinic acid isomers) were identified by LC-NMR-MS techniques.

5 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Plenary Lecture

6 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication

Oral Communications The Bioconversion of Native Starches

Vladimir V. Aksyonov, Anastasia V. Maximenko

Institute of the processing of agricultural products SB Russian Agricultural Academy, Novosibirsk, Russia, e-mail: [email protected]

The technologies of starch processing to starch products with purpose of their intensification developed by the variation and modification of catalysts from mineral acids to enzymatic agents. At the same time electrophysical, hydrodynamic and other effects as the ways of the intensification of hydrolysis processes which are used in different branches of chemical industry are attended not enough. The usage of various physical effects allows to increase the speed of biochemical reactions and obtain the results which are inaccessible by usual technologies. The intensification of production processes by the physical and biochemical effects is directed to the increasing of cost-effectiveness as a result of purposeful influence on equipment capacity, manpower and material costs saving, and improvement of quality indices of products. Physical effects such as mechanical, acoustical, hydrodynamic, and electromagnetic and others are power and leads to the changes of the system properties and condition. In this connection the problems solving in the process of the intensification can be considered as similarly to the problems of optimal control. It was shown that the usage of acid fraction of electroactivated water in the processes of bioconversion of various types of starch – corn, wheat, rye, and potato – allows one to shorten the process duration half as long. The usage of hydroimpulsive effects at the preliminary stage of starch processing allowed one to increase the bioaccessibility of starch granules for enzymatic hydrolysis, decrease the temperature of gelatinization of starches and power consumption of the whole process.

7 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Chemical Constituents of Hammada elegans (Bunge) Botsch and Their Antimicrobial Activity

Nabila Abduaziz Al-Jaber

Chemistry Department, College of Science (girls sections), King Saud University, Riyadh, Saudi Arabia Fax: 00699 14772245, e-mail: [email protected]

The phytochemical investigation of Hammada elegans (Bunge) Botsch; family Chenopodiaceae, afforded three flavonoids. They were quercetin and quercetin 7-O- rhamnoside and Quercetin-4'–methoxy-3–fructo–rhamnoside and 5 coumarins identified as scopoletin, scopolin, umbelliferone, xanthotoxol, and esculetin. The identification of these compounds was achieved by m.p., UV, EI-Mass, 1H NMR, 13CNMR, and co-cromatographed with authentic references. Quantitative estimation of total flavonoids using spectrophotometric method as will as mineral analysis were carried out. The cytotoxic activity of the ethanolic extract against human tumour cell lines U251 and MCF7 (brain tumour and breast carcenoma cell lines, respectively) proved no cytotoxic activity. The anticoagulant activity on rates showed a significant effect.

References 1. The New Encyclopedia Britanica, Mascropedia Benton publisher, Chicago 1974, 5th ed., vol. 6,63 2. Trease, G. E. and Evans, W. C. Pharamacognosy 1983, Bailliere Tindall, London, 12th ed. 3. Ageel, A. M; Mossa, J. S. M. ; Al-Yahya, M. A. and Al-Said, M. S. Saudi Plant Used in Folk Medicine 1987, King Abd-Elaziz City for Science and Technology (KACST), 93. 4. Loutfy Boulas. Flora of Egypt Checklist 1995. Al-Hadara Publishing, Cairo, Egypt. 5. Ajabneor, M.A.; Al-Yahya, M. A.; Tariq, M. and Jayab, A. A. Fito teropia 1984, vol. LV, 107. 6. Al Muhtadi, F. J.; El-Sherei, M. M. ; Shokah, A. A. and Awaad,.A. S. “Proceeding of XXII Conference of Pharmaceutical Sciences” 1992, P. 255-274. 7. Sandberg,F.; Michel,K. M.; Haglid, F. and Norim, T. Acta Pharm. Suecica 1967, 4, 97. 8. Sandberg,F. “Biochem, Physiol. Alkaloids, INT symp1969, 4, 177, through C. A. 9. Romero, M. A.; Wilson,S. J. ; Blunden, G.; Yong,H. M. ;Curatat. A. C and Bashir, R. A. Biochem. Syst. Ecol 1998, El Sevier Science Ltd., 26 (5), 535-543. 10. Stahl, E. Thin-Layer Chromatography 1969, 2nd ed., George Allon and Unwirid, London, Springer Berlin. 11. Geissman,T. A. Chemistry of flavonoids Compounds 1962, The Macmillan Company Ltd., New York. 12. Skehan, ‘J. Natl. Cancer. Inst.’ 1990, vol. 82, p 1107-1112. 13. Mabry;T. J. ;Markham K. R. and Thomas, M. B. The Systematic Identification of Flavonoids 1970, Springer Verlage, New York, Heidelberg, Berlin. 14. Murray, R. D. ;Mendez, J. and Brown, S. A. The Natural Coumarins, Occurrence, Chemistry, and Biochemistry 1982, John Willey and Sons Ltd., Chichester, 35. 15. Chalteijee, D. K. and Sen, K.. Tetrahydron Lett. 1969, 59,5223. 16. R. H. Goedwin and B. H. Pltock. Arch. Biochem. Biophys 1954., 49, 1. 17. Ling, Y. K.; Zhi, M. X. and Ting, R. Z.. Phytochemistry 1996, 41 (5), 1423-1426. 18. Harborne, T. B.; Mabry, T. J. and Mabry, H.. The Flavonoids 1975, Chapman and Hall, London.

8 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Synthesis and Biological Activity of New Derivatives of Mono- and Sesquiterpenoides of Essential Oils

Gayane A. Atazhanova

JSC “International research and production holding “Phytochemistry”, 100009, Karaganda, Kazakhstan Fax: (7212) 43-37-73 E-mail: [email protected]

The urgency of researches in the field of chemistry mono- and sesquiterpenoides of essential oils is caused by the practical importance of this class of natural compounds as accessible and renewed raw material for the obtaining of biologically active compounds. Objects of our research are monoterpenoides, such as ∆3-carene, α-and β-pinenes, camphene, artemisia ketone, pulegone, limonene, carvone, 1,8-cineol, camphor and sesquiterpenoides – chamazulene, buddledine C. We used the various approaches o t introduction nitrogen-, haloid- and oxygen-containing groups in molecules of mono- and sesquiterpenes, thus it is obtained more than 50 new derivatives. The initial terpenes and their derivatives are tested on antimicrobial, antifungal and cytotoxic activities. Among the studied samples of mono- and sesquiterpenoides and their derivatives, oxime cycloartemisia ketone has shown comparatively high bacteriostatic effect against all investigated test-strains. While cycloartemisia ketone has the moderate antimicrobial action in relation to Staphylococcus aureus, it has not shown activity to the rest of the test-strains. Artemisia ketone is active only in relation to gram-positive microorganisms Staphylococcus aureus and Bacillus subtilus. The gram-negative microorganisms were not sensitive to this substance. Thus, dinitropyrazol derivative of cycloartemisia ketone has shown antibacterial effect both to the gram-positive bacterium Staphylococcus aureus, and to gram-negative – Escherichia coli. Thus, cyclization of artemisia ketone leads to the change of spectrum of antibacterial activity hereby at a former level the activity is kept to Staphylococcus aureus and is not noted an effect concerning Bacillus subtilus, but it is found out in relation to Escherichia coli. The comparative analysis of antifungal activity of initial terpenoides and their derivatives testifies that the antimycotic properties are expressed weakly or they are absent. The chlorderivative of santoline alcohol and oxime buddledine show a weak antifungal activity concerning Candida albicans. Result of bioscreening of monoterpenoides and their derivatives on Artemia salina (Leach) has allowed to define the expressed cytotoxic activity in a number of the investigated monoterpenoides and their derivatives at sabinol and sabinilacetate. Thus, the made complex of researches has shown the availability of search and development of new antimicrobic preparations, first of all, in a number of nitrogen-containing derivatives mono- and sesquiterpenoides of essential oils.

9 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Phytochemical Study of Cyperus alternifolius L. as a hepatoprotective Plant Against Carbon Tetrachloride-Induced Hepatotoxicity in Rats

Amani S. Awaad

Chemistry Department Faculty of Sciences, King Saud University, Riyadh. KSA. P.O. Box 22452, Riyadh Fax: +96614772245 E-mail: [email protected]

Phytochemical screening for Cyperus alternifolius L. showed that this plant contains carbohydrates, glycosides, flavonoids, sterols and/or triterpenes as well as proteins and/or amino acids, free sugars were detected in the alcoholic abstract of this plant and identified as Rhamnose, glucose, arabinose, galactose, sucrose, glucouoronic acid and galactouoronic acid. Upon the investigation of lipid content of Cyperus alternifolius L. α-amyrine and β-sitosterol were isolated from the uns!aponifiable part and 14 fatty acids were detected and varied in their concentrations. Amino acids (free and compound) and Protein were studied, their percentage were calculated in addition to protein (found to be 9.57%). 8 phenolic compounds were isolated for the first time from this plant and identified as esculetin, umbelliferon, imperatorin, psoralen, xanthotoxin, quercetin, rutin, gallic acid using different spectroscopic analysis such as 1H-NMR, 13C-NMR DEPT, COSY, HMBC, HMQC and mass spectrometer. Hepatoprotective activity of the total ethanolic extract of the plant in doses of 50, 100 & 200 mg kg–1 and its successive extracts (Etherial, Chlororformic, Ethyl acetate, Butanol, Fat & chlorophyll) in a dose of 10 mg kg–1 was evaluated using hepatic injury model induced by

CCl4. The total and successive extracts were orally administered to different groups of rats for

9 consecutive days. Thirty min after the last dose, a single dose of CCl4 was injected subcutaneously to rats to induce hepatotoxicity. After 48 h of CCl4 injection, blood samples (2 ml) were collected from each rat and serum was separated to be used for biochemical assay.

Subcutaneous injection of CCl4 to rats induced liver injury as indicated by a significant exaggerated increase in the levels of AST, ALT and ALP enzymes, bilirubin, cholesterol, and triglyceride while total proteins and albumin were significantly reduced. Oral administration of the total extract in doses of 100 & 200 mg kg–1 and the successive extracts (chloroformic –1 and Fat & chlorophyll) in a dose of 10 mg kg to rats prior to CCl4 significantly improved these levels in serum toward the range of the control values.

10 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Flavonoids and Polysaccharides of Larch Wood, Isolation and Purification Technology and Experimental-Industrial Production Creation

V.A. Babkin

Laboratory of Wood Chemistry, A.E. Favorsky Institute of Chemistry Russian Academy of Sciences Siberian Branch, Favorsky St., #1, Irkutsk, 664033, Russia

Larch wood contains up to 4.5% of flavonoids, which are presented by one-type chemical structure compounds with the dominating content (more than 80%) of bioflavonoid dihydroquercetin (DHQ). The technology of obtaining of this valued biologically active compound was elaborated in the Laboratory of Wood Chemistry of Irkutsk Institute of Chemistry, Russian Academy of Sciences, Siberian Branch, in the 90-ies of the twentieth century. The phytopreparation Diquertin, containing more than 90% of DHQ, has been produced on basis of larch wood extractive substances. Diquertin, as a preparation providing capillary resistance recovery, possessing anti-inflammatory and anti-edematous activities, exerting positive influence upon the functional condition of liver, is recommended for pathogenetic therapy of bronchopulmonary disease and ischemic heart disease, instable angina and supraventricular arrhythmia. Arabinogalactan (AG) represents oneself as a unique dissoluble natural polysaccharide, possessing immunomodulating and prebiotic activities, which application promotes considerable invigoration of gastrointestinal tract, especially large intestines. The technology of isolation of AG from larch wood, where its content amounts to 15%, has been developed. By now, the technologies of obtaining of larch wood bioflavonoid dihydroquercetin (DHQ) and larch wood natural polysaccharide arabinogalactan (AG) are brought to the industrial level. With the purpose of obtaining of the new effective phytopreparations from the inexpensive renewable raw material, the complex wasteless processing technology of the biomasses of Siberian larch and Gmelin’s larch has been developed. The individual obtaining technologies of wood and bark extractives have been worked out. All of the extracted substances preserve their native properties and possess high biological activities. The method of isolation of phytocomplex from larch bark, the basic monomeric flavonoids of which are DHQ and, the specimen of the new class of flavonoids – spirobiflavonoid larixinol, has been developed. On basis of the characterized phytocomplex, biologically active food additive Picnolar has been developed and has undergone the state registration. Picnolar is a source of natural antioxidants. It possesses the expressed hepatoprotective properties.

11 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Anticoagulative Effect of the Preparation of Serine Proteases Inhibitor from Hepatopancreas of King Crab (Paralithodes camchaticus)

Maria V. Balashova1, Galina N. Rudenskaya2*, Ylia A. Rudenskaya1, Viacheslav A. Isaev3, Lydmila V. Lytova1

1 MSU, department of biology, Moscow 2 MSU, department of chemistry, Moscow, e-mail: [email protected], [email protected] 3 Trinita company, Moscow

Thrombosis and thromboembolism are one of the most grave and wide-spread cardiovascular diseases. The treatment of these diseases usually includes injections of thrombolytic agents, which trigger the dissolution of blood clots. Another approach deals with anticoagulant agents, such as thrombin inhibitor and thrombin activator inhibitor. Our investigation is aimed to expand the range of anticoagulant drugs. We found out that the preparation of serine protease inhibitor, obtained from hepatopancreas of King Crab, has an obvious anticoagulative activity towards human blood plasma. The research was carried out via tromboelastography. It is established that the preparation of inhibitor itself redoubles the prothrombin time during clot formation; the coagulation index decreases by 34%, maximum amplitude decreases by 12%, the density of clot decreases by 17%. This indicates on good anticoagulant activity. In case of pre-incubation of preparation with blood plasma (30 min), the effect grows: reaction time increases by 61% while the coagulation index lowers by 79%, maximum amplitude lowers by 24%, clot density lowers by 32%. Furthermore, the speed of fibrin clot formation is slowing considerably. This tendency increases in case of pre- incubation of the inhibitor preparation with blood plasma for 60 minutes before recalcification. Heparin, being an anticoagulant agent reduces thrombus density by 35% and coagulation index by 51%. Though, itself heparin doesn’t affect considerably the reaction time and maximum amplitude, which lowers only by 26%. In case of combined action of heparin and inhibitor preparation, there is an increase of prothrombin time by 81% and a decrease of maximum amplitude by 82%. Moreover, significant lowering of clot density (by 88%) and coagulation index (by 94%) are observed. This indicates on a weaker fibrin clot formation as well as on a dramatic hipocoagulation. We suggested that heparin is a cofactor of he t examined inhibitor and operates following the same mechanism as with antithrombin III. In case of pre-incubation of heparin with the preparation of inhibitor, the process of blood clotting is completely suppressed. According to the obtained results, we can assume that the preparation of serine protease inhibitor has a noticeable anticoagulative activity, which increases dramatically in the combined action with heparin.

12 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Skeletal and Oxidative Transformations of Some Lead Triterpenic Acids for the Structure-Anticancer Activity Relationships Studies

M.V. Khudobko, L.R. Mikhailova, L.A. Baltina

Institute of Organic Chemistry Ufa Research Centre of RAS, prospect Oktyabrya, 71,450054, Ufa, Russia Fax: 7(347)2356066 E-mail: [email protected]

Plant’s triterpenoids are a widely distributed class of natural compounds that attract the great attention in the last decade due to the discovery of several highly potent antiviral and anticancer agents among derivatives of oleanolic, betulinic, moronic, ursolic, and glycyrrhetinic acids. Some pentacyclic triterpenes and their derivatives represent the great interest for the oncology as agents for the control of angiogenesis, invasion of tumor cells and metastasis and emergence as a new class of chemotherapeutics for the treatment of multidrug resistance tumors. Structural modifications of natural triterpenes are the perspective route to the construction of new highly potent anticancer agents. Typical among oleanane type triterpene synthetic derivatives are high cytotoxic 2-cyano-3,12-dioxo-18β-oleana-1,19-diene- 28-oic acid (CDDO), synthesized from oleanolic acid, and 2-cyano-3,12-dioxo-18β-olean- 1,12-diene-30-oic acid (β-CDODA), synthesized from Glycyrrhetinic acid (GLA) which possess peroxisome proliferators-activated receptor γ (PPARγ) agonist activity and ligands for this receptor are being developed as new anticancer drugs1. The main target of this work was synthesis of novel compounds with changed pentacyclic skeleton of molecules and oxidative transformations of some natural lead oleanane group triterpenic acids and their derivatives (11-desoxo-GLA, 18-19-dehydro-GLA, olean-9,12- dien- and olean-12-on-30-oic acids). Triterpenes A rings were transformed into 2-oxo-A-nor-, 2-cyano-3-oxo-olean- and 3-oxo-1-ens. Novel A-homo-A-aza-3-oxo- and C-homo-C-aza-3- oxo-derivatives were synthesized via Beckman’s rearrangements of corresponding 3-keto oximes. Synthesis of 3,12-dioxo-olean-1,9-dien-30-oic acid was carried out based on the 3-O- acetyl-12-oxo-oleanan-30-oic acid. Convenient methods of synthesis of novel 1-en triterpene derivatives containing additional oxygen (oxo, epoxy, hydroxy, carboxy) and halogen (Br,

Cl) functions in the position 2 have been developed by using ozone, m-CPBA, H2O2 and PhSeCl. Structures of target compounds were established by NMR 1H and 13C experiments and LC/MS. The preliminary tests showed that some compounds with changed A and C rings inhibit the proliferation of human’s lung adenocarcinoma cells line A549 in vitro.

This work was supported by the RFBR grant 08-03-00366 and SS 1725.2008.3.

1 Chintharlapalli, S.; Papineni, S.; Jutoori, I.; McAlees, A.; Safe, S. Mol. Cancer Ther. 2007, 6, 1588-1598, and references cited therein.

13 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Seasonal Fluctuations of Phycobiliproteins and Sulfated Polysaccharides in Ahnfeltiopsis flabelliformis (Harv.) Masuda. (Rhodophyta, Phyllophoraceae)

Anna O. Barabanova1, Anna O. Kravchenko1, Irina M. Yakovleva2, Irina M. Yermak1

1Pacific Institute of Bioorganic Chemistry, Far-East Branch of Russian Academy of Sciences, Vladivostok, 690022, Russia, fax: 7(4232) 31-40-50, e-mail: [email protected] 2 A.V. Zhirmunsky Institute of Marine Biology Far-East Branch of Russian Academy of Sciences, Vladivostok, Palchevskogo 17, 690041, Russia, fax: 7(4232) 31-09-00

Red seaweeds are a source of many substances with a broad spectrum of biological activities. The cell wall of red seaweeds contains of a number water-soluble polypeptides and photosynthetic pigments – phycobiliproteins (phycoerythrin (PE), phycocyanin (PC), allophycocyanin (APC)), carotenoids and chlorophylls. A distinctive feature of phycobiliproteins is their high fluorescent intensity and, thus, they are widely used as fluorescent targets for molecular probes in medicine. Also, phycobiliproteins are used as a natural protein dye in the food and in the cosmetic industries. The main structural components of cell wall of the red algae are sulfated polysaccharides – carrageenans and agars. These polysaccharides are well known for their gelling properties, used in a variety of industrial applications. Red seaweed Ahnfeltiopsis flabelliformis (Phyllophoraceae) is widespread in the Seas of Pacific Coast. Seasonal fluctuations of polysaccharide and photosynthetic pigment contents in Ah. flabelliformis were investigated. The reproductive form (carposporophyte) of the algae was collected in Amursky Bay, the Sea of Japan, from February to September 2009. According to our results, the content of polysaccharides and photosynthetic pigments varied monthly. The highest content of polysaccharides in Ah. flabelliformis was observed in February and May, and their minimal amount was recorded in September. This polysaccharides contain the galactose, 3,6-anhydrogalactose and sulfate esters. It was shown, that phycoerythrin is a predominant compound in Ah. flabelliformis, while the ratio of phycocyanin/allophycocyanin fluctuated every month. The maximum amount of phycobiliproteins was observed from February to the first decade of April. The minimum level for all the pigments investigated was observed at the end of April and also in September. Our analysis shown, that UV-B and photosynthetically active radiation (PAR) were the main factors affected the seasonal content of phycobiliproteins in Ah. flabelliformis. The abundance of carotenoids and APC was mainly depended on water temperature and PAR, while no one factor influenced the content of chlorophylls. PAR was the main factor, affected the seasonal content of polysaccharides in Ah. flabelliformis.

14 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication St. John’s Wort Flowers Composition

Irina I. Bayandina1, Tatyana P. Kukina2

1 Novosibirsk State Agrarian University Dobrolubova st., 160, 63003, Novosibirsk, Russian Federation E-mail: [email protected] 2 Novosibirsk Institute of Organic Chemistry, Siberian Branch of the Russian Academy of Sciences 9 prosp. Acad. Lavrent’ev, 630090 Novosibirsk, Russian Federation E-mail: [email protected]

Hypericum perforatum (St. John’s wort) is well known as medicinal plant containing numerous compounds with a broad spectrum of biological activity. Constituents that have stimulated the most interest include the naphtodianthrones hypericin and pseudohypericin. Recently we investigated Hypericum perforatum extracts (HPE) by GLC-MS and HPLC. As a result we discovered some new constituents in St John’s wort at first1. There are differences in components of flower and aerial part extracts. The ratio of hypericin and pseudohypericin is constant (1:4) in whole aerial part and flowers, but sum of hypericins is more in flowers raw material. The terpenoid composition differs in flowers and aerial part: flowers include more sesquiterpenoids, squalen and methyl oleanolate in comparison to aerial part. Minor sterols and dehydroabietic acid are absent in St. John’s wort flowers. Methyl oleanolate may be marker for standardization of HPE. There are a lot of differences in lipid composition of flowers and aerial part extracts. Fatty acids of flowers are shorter than of aerial part components. There are practically absent the aliphatic hydrocarbons and alcohols. Polyprenols of Hypericum perforatum were isolated and identified from hexane extracts. Polyprenols of are composed of 8–12 isoprene units. Predominant polyprenol is component with 9 isoprene units. There are polyprenols in free and esterified form in St.John’s wort. Only polyprenyl acetates are found in the flowers. The relation of free to esterified polyprenols is 1:4 in aerial part collected on July and 2:1 in aerial part collected on August. These peculiarities of chemical composition St. John’s wort flowers have to influence on it’s medicinal uses.

1 Kukina, T.P.; Bayandina, I.I.; Pokrovsky, L. M. Chemistry of plant raw material, 2007, 3, 39-45.

15 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Cellulose from Cereal Waste and Miscanthus Biomass

Vera V. Budaeva

Institute for Problems of Chemical & Energetic Technologies, Siberian Branch, Russian Academy of Sciences, ul. Socialisticheskaya, 1, 659322 Biysk, Altai krai, Russia Fax: 8(3854)30-17-25 E-mail: [email protected], [email protected]

The sharp reduction in output of cotton and flax to produce cellulose of various designations has resulted in a search for new sources of non-wood cellulose. Particularly, cereal straw has been considered as a reliable source of biomaterials, including cellulose, due to its availability and cheapness despite low content of native cellulose up to 40–45% in the feedstock [1]. The grain husk (specifically oats, up to 37–40%), another sort of cellulose- containing cereal waste, is a more attractive feedstock for industrial processing because it normally accumulates at processing plants [2]. Miscanthus, grown on plantations of the Institute of Cytology and Genetics SD RAS in 2008, has become a new cellulose-containing raw material in Russia, with a native cellulose content of 50–55%. Results of studies on conditions of Miscanthus to cellulose conversion reported in a recent paper speak well for this sort of feedstock as a promising one amongst non-wood species with cellulose content below 55% [3]. This paper reports methods and conditions of deriving cellulose from cereal waste and Miscanthus biomass, physicochemical properties and characteristics of bleached pulp and cellulose chemical modification products as well as results of cellulose production from Miscanthus using a pilot setup at IPCET SB RAS.

1 R.C. Sun. Cereal Straw as a Resource for Sustainable Biomaterials and Biofuels: Chemistry, Extractives, Lignins, Hemicelluloses and Cellulose. Publisher: Elsevier, 2010, 300. 2 V.V. Budaeva et al. Obtaining technical cellulose from straw and cereals seed shells. Journal of Mountain Agriculture in the Balkans, vol. 12, no. 5, 2009, 1027-1039. 3 V.V. Budaeva, R.Yu. Mitrofanov, V.N. Zolotukhin, G.V. Sakovich. Conversion of Miscanthus sinensis. Polzunovskiy Vestnik, 2009, no.3, 328-335. 4 M.B. Jones, M. Walsh. Miscanthus: For Energy and Fibre. Publisher: Earthscan, 2001, 192.

16 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Biosynthesis of Astaxanthin in Heterologous Hosts by Marine Bacterial crtW and crtZ Genes

Seon-Kang Choi1 and Norihiko Misawa2

1Gangneung Science Industry Foundation, Daejeon-dong, Gangneung, Gangwon 210-340, Korea, 2Central Laboratories for Frontier Technology, Kirin Holdings Co., Ltd., i-BIRD, Suematsu, Nonoichi-machi, Ishikawa 921-8836, Japan Fax:82-33-648-21 E-mail:[email protected]

Astaxanthin has been shown to have inhibitory effect toward oxidation of low-density lipoprotein in human (Iwamoto et al. 2000), preventive effect of diabetic nephropathy in diabetic db/db mice (Naito te al. 2004), preventative effects against cancer or anticancer activities (Tanaka et al. 1994; Chen et al. 1999), and enhancement of immune response (Jyonouchi et al. 1995; Chen and Park 2004). The biosynthetic pathway of astaxanthin in bacteria was elucidated at enzyme and gene level by in vitro (Fraser et al. 1997; Fraser et al. 1998) and in vivo (Misawa et al. 1995) studies. We compared the catalytic functions of various enzymes for astaxanthin production in heterologous hosts (Choi et al. 2005, 2006, 2007). A complementation analysis was conducted using Escherichia coli transformants to compare the catalytic efficiency of bacterial CrtW and CrtZ from Brevundimonas sp. SD212, Paracoccus sp. PC1, Paracoccus sp. N81106, Pantoea ananatis (Erwinia uredovora 20D3), marine bacterium P99-3, and P450 monooxygenase (CYP175A1) from Thermus rhermophilus HB 27, and a CrtO derived from Rhodococcus erythropolis strain PR4 and Synechosistis sp. PCC6803 for astaxanthin production in E.coli. The results obtained up to the present clearly suggest that the bacterial crtW and crtZ genes are a combination of the most promising candidates for developing recombinant hosts that produce astaxanthin. We have succeed in producing more than 0.5% plant (dry weight) astaxanthin (more than 70% of total carotenoid) in tobacco leaves by expressing both CrtW and CrtZ genes Brevundimonas sp. SD212. Moreover, the total carotenoid content in the transplastomic tobacco plants was 2.1-fold higher than that of wild-type tobacco.

17 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Optimization of Gelatin-Gum Arabic Complexed Microcapsules for Encapsulation of Allyl Isothiocyanate

Zhi-Qiang Zhang1, Cheol-Ho Pan2, and Donghwa Chung1

1Department of Marine Food Science and Technology, Medical and Bio-Materials Research Center, Gangneung-Wonju National University, Gangneung 210-702, Korea, Fax: 82-33-640-2347, E-mail: [email protected] 2Natural Products Research Center, KIST Gangneung Institute, Gangneung 210-340, Korea

Allyl isothiocyanate (AITC), a food flavoring agent possessing high chemopreventive and antimicrobial activities, was microencapsulated by gelatin-gum arabic complexation, in order to overcome the limitations of using AITC in food and nutraceutical systems, caused by its strong pungency, poor water-solubility, and high reactivity with nucleophiles. The effects of eight encapsulation process parameters were investigated on the encapsulation efficiency (EE), size distribution, and morphology of the microcapsules using a screening design. The significant parameters were further optimized for EE using response surface methodology, and the release of AITC from the optimized microcapsules was investigated in vitro. The microencapsulation was performed with AITC-containing olive oil as a core, gelatin and gum arabic as wall materials, Twin 80 as a surfactant, and tannic acid as a non-toxic cross-linker. An 8-factor-12-run Plackett-Burman screening design at two levels was generated with eight process parameters: total polymer concentration (X1, 14%), olive oil concentration (X2,

24%), AITC concentration in olive oil (X3, 2.55.0%), surfactant concentration (X4,

0.020.10%), cross-linker concentration (X5, 0.30.7%), cross-linking time (X6, 520 min), o ultrasonic power (X7, 72168 W at 20 kHz), and ultrasonic temperature (X8, 4050 C). A three factor, five-level central composite design was employed for the optimization. The release of AITC from the optimized microcapsules was investigated in a simulated gastric (pH 1.2) and intestinal (pH 7.5) fluids with or without digestive enzymes. The analysis of variance demonstrated that the EE was significantly affected by X1, X2, and X3, the diameter by X1, X2, X4, and X5, and the span by X1, X2, and X4 (p < 0.05). The shape and dispersibility of the microcapsules were also notably affected by the parameters. The statistical optimization showed that the EE could reach its highest value of 89.50% when X1 = 4.5%, X2 = 6.5%, and

X3 = 5.9%, at which the EE of 86.97% was obtained experimentally. The in vitro release study revealed that the release became more sustained with the increase of tannic acid concentration but became faster by the presence of pepsin and pancreatin. The degradation of microcapsules in the simulated fluids was confirmed by examining their morphology and size distribution.

18 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Anticarcinogenic Activity of Aaptamine and Its Derivatives Isolated from Marine Sponge Aaptos sp.

Sergey A. Dyshlovoy, Sergey N. Fedorov, Larisa K. Shubina, Valentin A. Stonik

Pacific Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Prospect 100 let Vladivostoku 159, 690022 Vladivostok, Russia Fax: (7-4232) 314050 E-mail: [email protected]

Marine sponges are a rich source of natural compounds possessing potent biological activities and belonging to various structural groups. The bioactivity-guided fractionation of the ethanol-aqueous extract of the marine sponge Aaptos sp., collected near the coast of Vietnam, yielded seven substances which possess cytotoxic anticancer properties and belong to the aaptamine structural group. Among them there are five previously known alkaloids aaptamine (1), 9-demethyloxyaaptamine (2), isoaaptamine (3), 3-(isopentylamino)-9-demethyloxyaaptamine (4), 3-(phenethylamino)-9- demethyloxyaaptamine (5) and two new alkaloids -(N 3-morpholinyl)-9- demethyloxyaaptamine (6) and 3-(methylamino)-9-demethyloxyaaptamine (7).

OCH3 OCH3 OCH3 OCH3 OCH3 OCH3 OCH3 CH3 O O HO O O O O

CH3 HN N N N N N N

N N N N N N N

NH (CH2)2 NH (CH2)2 N NH (1) (2) (3) (4) (5) (6) (7) CH3 CH3 CH3 O

Anticancer properties of the isolated substances 1-7 were further studied using the MTS- method for cytotoxicity evaluation, the soft agar method and JB6 Cl41 P+ tumor promotion sensitive cells for the study of the cancer-preventive activity, flow cytometry for apoptosis detection, the luciferase method for the evaluation of basal AP-1, NF-B, or p53 nuclear factor – dependent transcriptional activity. The results showed that the substances 1-7 possess low or moderate cytotoxic activity against tumor and normal cell lines, and strong or moderate cancer-preventive activity in doses 4-20 times less then cytotoxic. Flow cytometry using Annexin V–FITC and propidium iodide PI double staining demonstrated that all the substances studied started to induce early apoptosis at noncytotoxic concentrations. We found also that most of the compounds studied inhibit p53-dependent transcriptional activity at the active concentrations. This important finding demonstrated that apoptosis induced by these compounds might be p53-independent and therefore these substances may be used for the treatment of the p53-deficient cancer cell lines (such as HL-60), which are resistant to the anticancer chemotherapy using a number of medicines. At the same time our results demonstrated that compounds 1-3, 5 activate basal AP-1- and NF-B-dependent transcriptional activity indicating that these nuclear factors may be involved in the anticancer effects induced by aaptamine (1) and its derivatives 2, 3, 5. This study indicates that aaptamine-like compounds might be perspective as the cancer- preventive drugs.

19 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Sulphated Polysaccharides of Brown Seaweeds. History and Modern Day of the Investigation

Svetlana P. Ermakova

Pacific Institute of Bioorganic Chemistry, FEB RAS 159 100-letya Vladivostoky 690022, Russia Fax: 8(4232) 310705 E-mail: [email protected]

Various species of marine algae occur in the ocean, they are classified into three divisions: Rhodophyta (red algae), Phaeophyta (brown algae), and Chlorophyta (green algae). The main constituents of algae are polysaccharides which classified into two groups: storage and structural. Sulfated polysaccharides are a type of structural polysaccharide. Red and green seaweeds contain galactans and ulvans, respectively. Fucoidans present structural polysaccharides of brown algae. The most of fucoidans contain fucose as one of the constituent sugars. But in present time there are known fucoidans with ratio of fucose and galactose as 1:1, ascophyllans are xylofucoglucuronans with large proportions D-xylose and uronic acids. The amount of fucoidans in a brown alga varies by species of seaweed, place of location, and season. The fucoidas from Pelvetia canaliculata comprise from 18% to 24% of the dry weight of the algae, whereas in Durvillea potatorum the proportion of polysaccharides is very small (0% to 2%). Isolation and purification of fucoidas from algae are carried out through the several steps: drying and milling of algal materials; extraction of low-molecular-weight substances with extracting agents; extraction of fucoidans with dilute acid solution from residues of algae, and purification of sulphated polysaccharides from the extracts by precipitation with ethanol or fractionation on ion exchanger columns. It is necessary to chouse the conditions under which the maximum yield will be obtained. The early works on chemical composition of fucoidans in brown algae were done by Kylin in 1913. Fucose-containing sulfated polysaccharides are highly heterogeneous in both monosaccharide composition and molecular structure. They all comprise a family of heteromolecules based on L-fucose, D-galactose, D-xylose, D- glucuronic acid, and D-mannose. In spite of the fact that fucoidans have high variation in composition, their structure has been studied. However, the structure is still incompletely characterized because of the apparent complexity of the polysaccharides and usually only partial information on their structures can be obtained by NMR spectroscopy. Controversial data may be found in the literature, even about the structure of the most carefully studied fucoidan from F. vesiculosus. Fucoidans from brown algae have broad spectra of biological activity. They have been extensively investigated for their anticoagulant and antitrombotic activity. The subjects of investigations are antivirus, antitumor and immunomodulatory activities. Lots of studies show that fucoidan presents significant antioxidant activity in experiments in vitro. It can also remarkably reduce blood lipids. Anticomplementary and anti-inflammatory activities are also attractive for studying. The research progress on the structure and bioactivity of fucoidan and the relationships between structure and bioactivity will be presented.

20 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication New “Green” Way to Epoxydes of Terpenes

Vladislav V. Fomenko, Oleg V. Bakhvalov, Sergey S. Laev, Nariman F. Salakhutdinov

N.N. Vorozhtsov Institute of Organic Chemistry, 9 Ac. Lavrentjev Avenue, Novosibirsk, 630090, Russia E-mail: [email protected]

Epoxidation of olefins is convenient method of their activation and functionalisation, and frequently also the first stage of the industrial processes. On our opinion, most promising method of the scale processing of forest terpene raw material (turpentine) appears the way, which assumes the initial epoxidation of the terpene raw material. However, the classical methods of processing raw material require the large number of expensive and often dangerous reagents necessary for conducting the synthesis. This circumstance specifies, in particular, the formation of the large number of dangerous wastes. Often, the amount of waste at the synthetic stage exceeds a quantity of the obtained product [1]. We developed the new, cheap, patently clean and ecologically safe method of the epoxidation of a number it is terpene by aqueous hydrogen peroxyde [2]. Using mentioned method we successfully epoxidized β-pinene, limonene, 3-carene. The modification of the developed method makes it possible to conduct the selective oxidation of α-pinene in the turpentine to -pinene oxide and thus to combine the chemical stage of obtaining epoxies with the separation of turpentine into the individual substances. This route of -pinene oxide preparation from turpentine is more effective than expensive and diffucult separation of - pinene from turpentine followed by epoxidation.

1 R.A. Sheldon, H. van Bekkum. Fine Chemicals through Heterogeneous Catalysis 2001.Wiley-VCH P. 4.

2 Bakhvalov O.V., Fomenko V.V., Salakhutdinov N.F. Pat. RU №2005140845/04, №2006138540/04(041991).

21 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Plant Coumarins of Some Apeaceae Species Growing in Siberia and Mongolia

J. Ganbaatar1, E.E. Shults2, D. Mungunnaran1, A. Tuvshintugs1, G.A. Tolstikov2

1 Institue of Chemistry and Chemical Technology, Mongolian Academy of Sciences 2 N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, SB RAS, Novosibirsk, Russia

Coumarins are naturally occurring compounds and serve as source for producing of potential medicinal preparations. Plants of Apeaceae family widely spread in Siberia and Mongolia [1]; they produce a lot of linear and angular pyrano- and furocoumarins. In this report is described results of works concerning to investigation of chemical compositions of some plants belonging to genus Peucedanum and Phlojodicarpus. It has been also studied methods for isolation of valuable metabolites from above mentioned plants. Also in report is considered the methods for isolation of the most valuable metabolites found in above mentioned plants. We have revealed qualitative variability of plant’s coumarin composition due to places where they grow. In particular have been discussed:  the composition of the main coumarin metabolites of Phlojodicarpus sibiricus, Ph.turczaninovii [furo- and pyranocoumarins: peucenidin, oroselon, dehydrolomatin, (-)-(R)-decursinol, bukhtarmin and sacsdorfin (the total content of coumarins is 2.2-3.2 %)];  the composition of the main coumarin metabolites of Peucedanum morisonii; P. decursivum, P. terebinthaceum, P. Praeruptorum (peucedanin, decursin, decursidinols, pterixin, praeruptorin, praeroside). Considerable attention has been given to the study of questions concerning to optimization of methods for isolation of following coumarins for providing of their synthetic transformations. These are: (-)-cis-khellactone (Phlojodicarpus sp.) and (+)-trans-khellactone (P. praeruptorum), these compounds are very interesting for creating of antiviral agents [2]; peucenidin and (-)-(R)-decursinol (Phlojodicarpus sibiricus). The latter compounds serve as inhibitor of acetylcholinesterase [3].

1 V.I.Grubov, Key to the vascular plants of Mongolia, 1982, 192-193. 2 Zhang, Q.; Chen, Y.; Xia, P.; Xia, Y.; Yang, Z.-Y.; Yu, D.; Morris-Natschke, S.L.; Lee, K.-H. Bioorg. Med. Chem. Lett., 2004, 14, 5855. 3 Kang, S.Y.; Lee, K.Y.; Sung, S.H.; Park, M.J; Kim Y.C. J. Nat. Prod., 2001, 64, 683.

22 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Study on Chemical Constitutes of the Volatiles from Dracocephalum nutans L.

Jenis Janar1, G.Sh. Burasheva1, Haji Akber Aisa2

1Chemistry Department of Al-Farabi Kazakh National University, Almaty, 050012, Republic of Kazakhstan. E-mail: janarjenis @ mail.ru; 2Xinjiang Technical Institute of Physics and Chemistry, Urumchi, 83001, China

The genus Dracocephalum, which belongs to the Lamiaceae family, in Kazahstan 49 genus, 247 species of plants are growing that belong to family Lamiaceae and genus Dracocephalum have more than 20 species. Various species of Dracocephalum have medicinal values, e.g. Dracocephalum parviflorum has been used to treat diarrhoea in children. The leaves are febrifuge and ophthalmic. A cold compound infusion has been used in the treatment of fevers and headaches. Externally, it has been used as an eyewash. In Kazakh traditional medicine Dracocephalum nutans L. has been used in the treatments of bronchitis, bronchial asthma, brohepatitis and urethritis. In this study an attempt has been made to investigate the chemical constituents of volatile oils obtained from the aerial parts of Dracocephalum nutems L. grown in Altay mountain (Altay city, Xinjiang Automonic Region of PRC) for the first time. The chemical constitutes of volatiles of Dracocephalum nutans L. by method of Water steam distillation was analyzed by GC-MS. And relative content of each component was determined by area normalization. Fifty three peaks were separated and forty nine compounds were identified from Water steam distillation method, the major constituents are [1aR-(1aα.,4aα.,7β,7aβ,7bα)]-decahydro-1,1,7- trimethyl-4-methylene-1H-cycloprop[e]azulen-7-ol (14.23%), caryophyllene oxide (11.3%), 3,7-dimethyl-1,6-octadien-3-ol (7.69%), myrtenyl acetate 5.67%), ( 1,2,3,4,4α,5,6,8α- octahydro-7-methyl-4-methylene-1-(1methylethyl)-naphthalene (5.29%), (-)-spathulenol (4.76%), 1-methoxy-4-(1-propenyl)-benzene (4.73%), dehydro-1,1,4,7-tetra-[ethrenthyl]-4aH- cycloprop[e]azulen-4a-ol (4.65%), (E)-3(10)-caren-4-ol (4.42%), germacrene (4.23%), [1R- (1R,4Z,9S)]-4,11,11-trimethyl-8-methylene-bicycle[7.2.0]undec-4-ene (3.67%), 1,5,5,8- tetramethyl-[1R-(1R≠,3E,7E,11R≠)]-12-oxabicyclo[9.1.0]dodeca-3,7-diene (2.13%).

23 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Methodology of Cytostatic Drug Development on the Basis of Chromatographic Isolation and Hightroughput Screening of Plants Extracts

S.G. Klochkov

Institute of Physiologically Active Compounds of Russian Academy of Sciences, Chernogolovka, Russia Fax: (496)524-2525; e-mail: [email protected]

Throughout medical history, plant products have been shown to be valuable sources of novel drugs. Very known examples are the Vinca alkaloids, the taxanes, and the camptothecins. Natural products are the major source of chemical diversity that has consistently proven its value for the development of novel drugs even when compared with the powerful combinatorial chemistry techniques [1]. But only new high-throughput methods of screening natural compounds can effectively reveal new potential drugs [2]. In the present project the methodology of search of new biologically active compounds with cytostatic activity on the basis of a combination of methods of semipreparative HPLC, LC-MS/MS and high-throughput screening is offered. We have developed a novel high-throughput screen that is easily automated and compatible with high-throughput screening robotics. The new screen is quantitative and allows inhibitory potencies and concentration gradient to be determined. During chromatographic separation of extracts it is simultaneously formed two plates – deep-well plate for different types of biological screening and an analytical plate for LC-MS/MS investigations. The screening plate goes for complex testing on cytostatic, bactericidal activity, etc. The analytical plate after activity detection is investigated with the use of mass- spectrometry. Combination of these methods allows to reveal the most active compounds from plants with the use of very small starting plant material – for extraction we use only 25 gram of dried plant material. The efficacy of this new screen was illustrated by testing plant materials including more then 200 plant fractionated extracts and library of natural compounds. There were 36 active compounds among the library set. One very active extract was selected for bioactivity-guided fractionation resulting in the identification of compound Ld-12-475 as a potent cytostatic agent.

1 A.L. Harvey. Trends Pharmacol Sci. 1999, 20, 196 2 N.C. Gassner et. al. J Nat Prod. 2007, 70, 3, 383

24 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication A Developed Technology of Fir Needles Complex Processing

Tatyana P. Kukina, Tamara A. Vaganova, Eugeny V. Malykhin, Sergey A. Popov, Andrey M. Chibiryaev

Novosibirsk Institute of Organic Chemistry, Siberian Branch of the Russian Academy of Sciences, 9 prosp. acad. Lavrent’ev, 630090, Novosibirsk, Russian Federation E-mail: [email protected]

The genus Abies of family Pinaceae consists of 51 species. Fir is used for building materials, roof shingles, interior paneling, weaving looms, furniture, shipping crates, firewood and Christmas trees. Fir pulp is used extensively for making high-grade wrapping paper. The literature data on the genus of Abies revealed that were isolated more than 400 compounds. The chemical constituents are mostly terpenoids, flavonoids, lipids, lignans, phenols, polyprenols, dolichols, glicosides, polysacharides, steroids, and others. About 100 compounds are triterpenoids. They are actually only two structures of lanostanes and cycloartanes with trace of gammaceranes and hopanes. Different extracts from the leaves of Abies exhibited many kinds of activities, including insect juvenile hormone, antitumor, antibacterial, antifungal, anti-ulcerogenic, anti-inflammatory, antihypertensive, antitussive, and CNS activities. The most widely distributed species is Siberian fir Abies sibirica. It is one of the predominant trees of Siberian forests. The researchers of Novosibirsk Institute of organic chemistry (NIOCh) investigated the constituents of Siberian fir extracts. The result of this work was evaluation and identification more than 100 constituents including about 40 triterpenoids. Professor Raldugin patented biological active fraction of fir triterpenic acids as a high effective product for agriculture1. NIOCh developed the production of this product in industrial scale. Yield of this product is about 6 % of air dried raw material. The byproducts are neutral part of extract and raw material after extraction. This fact puts two main problems: raising the yield and utilization of wasts. Our researches led to patenting of some new effective products for agriculture and veterinary2, 3, 4. Whole conversion of raw material was developed up to 40% that releated with other schemes of fir raw material processing5. New types of bioactivities of these products were found at first6. Neutral part of extract also may be used for evaluation of oligoprenol fraction with broad scale of bioactivity. The method of preparation and anti-influenza drug were also patented7, 8.

1 Chekurov, V.M.; Kolotygin, A.I.; Raldugin, V.A. et al. RU Patent 2025976, 1995. 2 Vaganova, T.A.; Kukina, T.P.; Malykhin, E.V. et al. RU Patent 2336889, 2006. 3 Popov, S.A., Kozlova, L.P., Kukina, T.P. et al. RU Patent 2348168, 2007. 4 Vaganova, T.A., Kolomnikova, V.I., Kukina, T.P. et al. RU Patent 2372930, 2008. 5 Vaganova, T.A.; Malykhin, E.V.; Kukina, T.P. et al. Chemistry for sustainable development, 2007, 15, 291- 308. 6 Um, B.H.; Kim, S.N.; Lee, W. et al. Chemistry for sustainable development, 2008, 1, 53-57. 7 Raldugin, V.A., Kukina, T.P., Tsytsenko, T.A. et al. RU Patent 2053992, 1996. 8 Safatov A.S., Kukina, T.P., Raldugin, V. A. et al. RU Patent 2189231, 2002.

25 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Mechanochemistry of Plant Materials

Oleg I. Lomovsky

Institute of Solid State Chemistry and Mechanochemistry of Siberian Branch of Russian Academy of Sci., Kutateladze str., 18, 630128, Novosibirsk, Russia E-mail: [email protected]

The moving force of plant materials mechanochemistry is the partial change of chemical industry base by bio renewable one. The unique natural resources of Russia must be utilized to alive as industrial country. Plant raw has a peculiarities in comparison with objects of traditional inorganic mechanochemistry: the cell structure defending the plant under mechanical and chemical damages, the heterogeneous chemical composition, 90% content of high strength and stable lignocellulose and cellulose biopolymers, the high reactivity of the most valuable biologically active components (hydrocarbons, polyphenols, acids, amines, glycosides) leading to the degradation under mechanical treatment. The difference of raw components mechanical properties is the supplementary factor defining the direction of chemical reaction of components under mechanical treatment. The cellulose component of raw plays the special role in mechanochemical transformation, it enhances the stability of complex organic molecules. Degradation kinetics of these molecules have the threshold character defining the optimal parameters of intensity and time of mechanochemical treatment. The optimal conditions of mechano enzyme processes are defined by stability and reactivity of enzymes in the presence of cellulose matrix. Mechanochemical reactions of hydrolysis, glycosilation, reglycosilation, soluble complex formation are used for the production of water soluble bio accessible products. The advantages of solid state mechanochemical technologies in relation to liquid phase ones are the enhanced yield and selectivity, the change of content of extracted mixtures of biologically active substances. The mechanically activated enzyme hydrolysis of cellulose and lignocellulose into soluble sugars is investigated as the first step of “second generation” bioethanol production from lignocellulose raw. The comparison is presented of Mechanochemical technology with European analog – DONG Energy technology. Some related problems of plant materials treatment into soluble sugars and possible mechanochemical solutions are discussed: the enzyme treatment for enhancing of plant material milling; possibility of plant raw enrichment and cellulose content increase via separation of cellulose (unwooden) and lignocellulose (wooden) plant tissues; the “wastes to wells” approaches for wooden fraction as disperse solid powder fuel and ethanol producing yeast biomass as source of fodder antibacterial additives for animals. The design trends of special mechanochemical equipment are also under scope, including vibration centrifugal mill-activators, roll mills with regulated impact intensity, conception of mobile mechanochemical modules on standard transport container base. So, the modern trends in high tech in “green chemistry” are presented: solid state “extraction” of biologically active substances from plant materials, mechano enzyme hydrolysis of lignocellulose into bio accessible sugars, enrichment of plant raw via separation of plant tissues.

26 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Some Studying Results of Kazakhstan Wild-Growing Plants and the Ways of Their Research

D.U. Korulkin, G.A. Abilov, R.A. Muzichkina

KNU named by al-Farabi, chemistry department, str.Karasaibatyr 95”a”, Almaty, Kazakhstan E-mail: [email protected]

In the state register of the USSR it has been described about 300 years of herbs and more than 500 medical products which were resolved to application in medical practice and industrial production. Since then, these lists annually are supplemented and extended among use of plants, especially with a complex of useful properties. The certain contribution to this direction was brought by researches of structure wild-growing Kazakhstan plants and phytopreparations on their basis. The flora of Kazakhstan includes more than 6000 kinds of plants from which more than 700 kinds are endemic. By this time the employees of chair has been studied more than 350 kinds from which 14 endemic, it is allocated in an individual kind and structure more than 400 connections, mainly polyphenolic character is completely established: flavonoids of various types, antraqinones, tannins, of the hydrolyzed and condensed types, polysaccharides, terpenoids, coumarins. In official medicine 21 plants are introduced. There are 9 phytopreparations and 14 medicinal forms are established on their basis. It became possible by means of complex chair researches of JSC «Himpharm» and medical profile institutes. The ways of series reception of the phytopreparations possessing antivirus, cicatrizing, radio protective, gepatoprotectorian, cholagogic, anti-inflammatory and antioxidant properties were developed and patented. For introduction in practice is offered following: 21 new Kazakh wild-growing herbs; the scientifically proved block-schemes of allocation and derivation of biologically active plant substances and manufacture processing of raw materials for phytopreparations; the advanced techniques of qualitative and quantitative definition of polyflans, polysaccharides, aminophenol acids, flavonoids, antraquions, tannins suitable for selection and standardization the quality of any medicinal vegetative raw material; technologies and regulations for manufacture of 9 substances with anti-inflammatory, cicatrizing, gepatoprotectorian, antivirus, cholagogic preparations at anti- inflammatory and antioxidant streams; technologies and regulations of medicinal forms manufacturing as ointments, syrups, tablets, etc.

27 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Contrastive Analysis of Composition and Bactericidal Activity of Various Fraction of Siberian Pine Essential Oil

Anton N. Narchuganov, Anna A. Gontova, Alexander A. Efremov

Siberian Federal University, 79 Svobodny Prospect, Krasnoyarsk 660041Russia Fax: 7 3912 44 86 25 E-mail: [email protected]

It is well known, that components of essential oils are characterized by a wide spectrum of influence on human organism: anti-inflammatory, anticancer, bactericidal and antioxidant activity. The aim of this study was to compare bactericidal activity and composition of various fraction of Siberian pine essential oil. Essential oil was produced by a steam distillation method. Duration of steam distillation process was 23 hours, and was received 5 fraction of essential oil, which differed on volatility of components. For the analysis of composition of the received essential oil used gas chromatography- mass spectrometry method. Identification of components was based on calculation of retention index (RI) and comparison of these indexes and mass-specters of individual compounds with library data1. Also, was defined bactericidal activity (minimal inhibitory concentration) of differ fraction of oil concerning bacteria Staphylococcus aureus and Klebsiella pneumonia. First fraction of essential oil (which was received in the first 5 hours after start of distillation) was consisted of 21 major components, basic of which was alpha-pinene (64%). The last fraction of essential oil was received in final period of steam distillation process (at 14 to 23 hours), and consisted of 49 major components, among which were discovered alpha- pinene (20%), beta-phellandrene (29%), delta-cadinene (9%). Minimal inhibitory concentration (MIC) of differ fraction of oil was differ also. So, MIC of first fraction of oil concerning bacteria S. aureus amount 5,31 µg/ml, and 2,55 µg/ml concerning bacteria K. pneumonia. MIC of last fraction of oil concerning both of bacteria S. aureus and K. pneumonia amount 1,33 µg/ml. Thus, various fraction of essential oil are differed on its componential structure and properties, and from the medical point of view last fractions of essence are more valuable.

1 Tkachev, A. Research of volatile compounds of plants; Ofset: Novosibirsk, 2008; pp 221-230.

28 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Doping agents and Oriental Herbs in Korea

Jae Keun Oh

Lab of Sports Medicine Korea National Sport University

Korean athletes popularly have taken oriental herbs as a ergogenic aids to improve their exercise performance and physical condition. But some sort of herbs was denoted to contain doping agents listed by World Anti-Doping Code of WADA. So Korean researchers have tried to find out doping agents in the oriental herbs by immunoassay and examine doping test among elite athletes. Doping Control Center (DCC) in Korea analyzed suspicious herbs and herbal prescriptions using established assay methodology for all WADA banned substances. The banned substances include stimulants, narcotics, anabolic steroid, beta blockers, diuretics and peptide hormones. As a result of research, some herbs such as Stephania Cepharanthra Hayata, Chelidonium majus L., Papaveris Fructus, Papaver rhoeas L., Strychnos nux-vomica L., Strychnos Ignatil Berg, Ephedra sinica Stapf, E. Equaisetina Bge, E. intermedia Schrenk et Mey, Pinellia ternata Breit, Sida cordifola L., Cannabis Fructus and herbal prescriptions which include prohibited herbs contained morphine, codein, cortisone, 17-oxycortico sterone, strychine, testosterone, ephedrine, cannabinol, androsterone and so on. And we made up a questionnaire about the use of herbs and traditional products supplements and doping test among elite athletes. 228 athletes of 14 events completed a questionnaire and DCC analyzed urine samples of 10 athletes using the multi-stratified cluster random sampling method. All of samples, the results of WADA anti-doping code test were negative into 9 items and 210 substances. Although some herbs contain prohibited substances and athletes were administrated safely, athletes have to increase the doping consciousness level.

29 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Synthesis and Cytotoxic Activity of 7,10-Dihydroxy-12H- benzo[b]xanthene-6,11-dione Derivatives Analogs of Bikaverin

Dmitry N. Pelageev, Alexandra S. Kuzmich, Victor Ph. Anufriev

Pacific Institute of Bioorganic Chemistry, Far-Eastern Branch of the Russian Academy of Sciences, 159 Prospect 100-letiya Vladivostoka, 690022 Vladivostok, Russian Federation Fax: +7 (4232) 314050 E-mail: [email protected]

Bikaverin (1a) is a red pigment with a wide-range biological activity. It was isolated from cultures of Gibberella fujikuroi, Fusarium oxysporum, and Mycogone jaapai. By this moment several methods of bikaverin synthesis are known. But all these syntheses are multistep and based on the use of reagents difficult of access. We found that benzopyranonaphthazarins 2 are substrates for synthesis of potential biologically active xanthones such as bikaverin and its analogs 1. Starting benzopyranonaphthazarins 2 were obtained by condensation of 2-hydroxynaphthazarins with 2-hydroxy- or 2-methoxy-benzaldehyde 2-bromo-3-hydroxynaphthazarins 3 and bromo derivatives 4 are the products of pyranonaphthazarins 2 bromination. We obtained other xanthenediones 5,6 by reaction of bromo derivatives 4 with different nucleophilic reagents such as acetone (enol form) and succinimide. Xanthenones 1b-f were obtained after treatment of compound 4 with water and following oxidation with air.

O HO R3 OH O R3 O HO OH O R3 2 R1 O R4 R1 O R4 Br R R1 O R4

1 R2 R5 R2 HO R R2 R5 O HO O R6 OH O H R5 O HO OH O R7 R6

1 4 2 3 5 6 O HO 1 2 1 4 2 3 5 6 1 R =R =OMe, R =R =R =H, R =Me (a); R2 3 R =OMe, R =H (a); 4 R =R =OMe, R =R =H, R =Br, R =Me (a); R1=R4=OMe, R2=R3=H, R5=Br, R6=Me (b); R1=R2=OMe (b); R1=R2=R4=OMe, R3=R5=H, R6=Me (b); 1 2 3 5 6 1 2 1 2 4 5 6 3 R =R =R4=OMe, R =R =H, R =Me (c); 1 R =R =H (c); R =R =R =R =R =H, R =OMe (c); HO R R1=R2=R4=R5=R6=H, R3=OMe (d); R1=R2=Me (d); R1=R2=Me, R3=OMe, R4=R5=R6=H (d); R7=Br; O HO R1=R2=Me, R3=OMe, R4=R5=R6=H (e); R1=R2=Cl (e) 5 R1=R4=OMe, R2=R3=H, R5=Br, R6=Me (a); 1 2 3 4 5 6 1 2 3 4 5 6 7 R =R =Cl, R =OMe, R =R =R =H (f) R =R =Cl, R =OMe, R =R =R =H (b), R =CH2Ac; 6 R1=R2=Me, R3=OMe, R4=R5=R6=H(a); 1 4 2 3 5 2 R =R =OMe, R =R =H, R =Me (a); R1=R2=R4=OMe, R3=R5=H, R6=Me (b), R1=R2=R4=OMe, R3=H, R5=Me (b); 7 R =N(COCH2)2 R1=R2=R4=R5=H, R3=OMe (c); R1=R2=Me, R3=OMe, R4=R5=H (d); R1=R2=Cl, R3=OMe, R4=R5=H (e) Furthermore, we established that 1d, 1e, 2e, 3a-d, 5a, 6b exhibit low or moderate cytotoxicity against HeLa tumor cells (IC50 = 16.9, 22.24, 27.46, 161, 130.4, 91.11, 15.3, 16.46, 21.28 ?M, respectively).

The work was supported by grant 10-III-B-05-092 from FEB RAS.

30 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Gene-Design for the Optimal Expression in Yeast of the Genetically Modified Constructions Including preS and S-genes of A and D Genotypes of Hepatitis B Virus

Svetlana V. Polyarush, Shakhnoz S. Azimova

Institute of Chemistry of Plant Substances, Mirzo-Ulugbek Str. 77, 100170 Tashkent, Republic of Uzbekistan Tel: (0998)2622099, Fax: (0998) 1206475, E-mail: [email protected], [email protected]

The main component of modern vaccines against hepatitis B (HBV) is recombinant S- protein, based on virus HBs surface protein. However, immunogenicity and specificity of this type of vaccines is rather low and almost 10-15% vaccinated population is prone to infection [1]. One of the attractive possibilities to increase immunogenicity is the use of preS envelope proteins. Should be noted, however, that the preS region is highly variable and with complicated structure in dependence on the HBV genotype [2]. Numerous heterologous proteins have been produced in the yeast genera Saccharomyces cerevisiae. The factors that drastically influence protein production in this system include: the expression cassette, mRNA 5'- and 3'-untranslated regions (UTR), translational start codon (AUG) context, A+T composition of cDNA, transcriptional and translational blocks, nature of secretion signal and endogenous protease activity [3]. In this work, we describe the expression of modified HBsAg in S.cerevisiae using the rational gene-design of the amplified DNA of HBV A-and D-genotypes from well characterized plasma samples. The design included the deletion of the protease sites, usage- codon changes with deletion of rare codon and truncation of the preS-region The yeast cells with recombinant constructs have been transformed and recombinant clones of yeast have been selected. The positive expression of the preS/SHBs constructs in the recombinant yeast by ELISA was observed.

1 Marwick, C.; Mitka, M. Debate revived on hepatitis B vaccine value. JAMA, 1999, 282,15-17. 2 Weber, B. Recent developments in the diagnosis and monitoring of HBV infection and role of the genetic variability of the S gene. Expert Rev. Mol. Diagn. 2005, 5, 75-91 3 Guthrie, C. and Fink, G. Guide to Yeast Genetics and Molecular Biology 1991, p. 933.

31 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Content of Alkaloids in the Different Parts of Sophora flavescens Soland

Denis V. Sandanov1, Natalia A. Pankrushina2,3

1Institute of General and Experimental Biology of SB RAS, Sakhyanovoi str., 6, 670047, Ulan-Ude, Russia 2Novosibirsk Institute of Organic Chemistry of SB RAS, Acad. Lavrentjev Avenue 9, 630090, Novosibirsk, Russia 3Novosibirsk State University, REC 008, Pirogova 2, 630090, Novosibirsk, Russia Fax: +7-3012-433034 E-mail: [email protected]

Quinolizidine alkaloids are the largest single group of legume alkaloids which appear to be restricted in distribution to species in the more primitive tribes of the Papilionoideae. These compounds are important due to their toxicity in humans and livestock as constituents of poisonous plants, and recent studies showed that some of them exhibit potentially useful pharmacological activities. Two types of quinolizidine alkaloids, matrine-type and pyridone quinolizidine bases, are found in Sophora species and can be used for the identification of Sophora species in commercial preparations. We began the study of alkaloids of Sophora flavescens Soland. This plant has been widely used for centuries in folk and Oriental traditional medicine. However, a long-term over-gathering of its roots and rhizomes has made this plant facing with the risk of being exhausted. This species nowadays become rare at the north-west part of distribution (Eastern Zabaikalye, Russia). One of the main tasks now is to find the way of its rational usage. The goal of our study was to analyze the dynamic of alkaloids in different parts of S. flavescens. Raw material was collected in 2000-2004 in the Eastern Zabaikalye. Results showed that the most part of alkaloids accumulated in the seeds (2.13-2.82%). S. flavescens has big seed productivity. Mass of 1000 seeds varies from 39.4 to 58.8 g and some of generative plants can have more than 300 seeds per one shoot. This data showed that seed reproduction is very important for this species. Content of alkaloids in the roots and rhizomes varies from 1.16 to 2.47%. The highest content of alkaloids was observed during the flowering period. This period is determined by active growth of rhizomes and it might be the main reason of such a phenomenon. Comparison of our data with climatic parameters showed that in humid years plants accumulated more alkaloids. Dry conditions reduced the growth processes, during dry years the small quantity of alkaloids was observed in the underground part of plants. The lowest content of alkaloids was observed in herb (0.25-0.29%). The content of alkaloids rapidly decreased in the herb of S. flavescens during fructification period. This dynamic observed can be explained by expending all plant resources to seed production. In conclusion, analysis of the dynamic of alkaloids in S. flavescens helped us to estimate the total accumulation of this group of biologically active substances in the plant and revealed some interesting biological peculiarities of this rare species.

32 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Synthetic Transformations of Alkaloids, Terpenoids and Coumarins of Siberian Flora for Search of New Pharmaceutics

Elvira E. Shults

Novosibirsk Institute of Organic Chemistry, SB RAS, 9 Acad. Lavrentyev ave, 630090 Novosibirsk, Russia E-mail: [email protected]

Synthetic transformations of natural product traditionally have played an important role in the discovery of new medicinal preparations. Today, drugs developed by the modification of natural compounds are engaged a leading position in the selling. Our laboratory has had a long-term focus on the chemistry of plant metabolites. The approach is to use of the direct synthetic transformations of accessible natural products followed by biological studies. The purpose of this communication is to review the methods developed in our laboratory for direct synthetic transformations of the above compounds isolated from Siberian plants. Two base strategies: the Diels-Alder reaction and cross-coupling reactions will be considered. The application of furan-type diterpenes and labdanoid furfuryl amides in the Diels-Alder reaction resulted in the synthesis of terpenoids containing heterocyclic fragments of 4-aza- 10-oxatricyclo[5.2.1.02,6]decene, 3-aza-10-oxatricyclo[5.2.1.01,5]-decene, 3-aza-10- oxatricyclo[5.2.1.01,5]decenone, and isoindolin-2-one. A number of heterocyclic systems were assembled/modified via Diels-Alder reactions of khrysenequinonecarboxylic acid or 7-[2- (2,6-dimethyl-6-vinyl-2-carboxy-cyclohexyl)ethyl]-7-isopropyl-1,4-naphthoquinones (obtai- ned from levopimaric acid) with various dienes. A logical evident route, based on the transition metal catalyzed reaction has been applied in a series of alkaloid transformations. Selective methods for brominating and iodination of diterpene alkaloid lappaconitine and morphinanes have been found. The Heck cross-coupling of 5-iodolappaconitine with ethyl (methyl) acrylates, 2-methyl-5-vinylpyridine, styrene and alkyl vinyl ethers afforded new olefinated derivatives. Derivatives of lappaconitine containing acetylenic substituents in the aromatic ring were prepared by the Sonogashira reaction. Oxidative coupling of acetylene compounds led to the formation bivalent ligands of the aconitane type. 1-Substituted morphinanes of new structural type were also obtained by the cross-coupling reaction. We have shown the utility of the Heck reaction to generate -arylidene--lactones with selectivity for the (E)-olefin geometry from eudesmane-type sesquiterpene lactones with - methylene--lactone substructures. The palladium-catalyzed amination was applied for the transformation of furocoumarins. In the results of pharmacological investigation of the new compounds in vivo are reveal:  highly active analgesics;  antiarrhythmic agents in the alkaloid series;  active anti-ulcer compounds in the sesquiterpene lactone series;  anti-viral and antiproliferative terpenoids.

33 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication A New Multifunctional Agents Synthesized on Natural Triterpenoid Platforms

I.V. Sorokina, T.G. Tolstikova, D.S. Baev, G.A. Tolstikov

N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry of SB RAS Acad. Lavrentjev av., 9, Novosibirsk, Russia e-mail: [email protected]

Natural triterpenoid platforms are an important scaffold for a new drug development. The secondary plant metabolites – ursolic and betulonic acid – are now successfully used for synthesis of non-toxic multifunctional substances, possessed a spread line of pharmacodynamic properties. These triterpenoid agents can regulate a wide range of dysfunctional pathways, and therefore may correct a different cell disorders, pre-malignant and malignant status. It is reported multifunctional triterpenoid agents may be useful to both prevention and therapy of cancer as well as to be synergistic with standard anticancer treatments. In present report we have tested in vivo more then twenty new derivatives of betulonic (BA) and ursolic (UA) acids and found some perspective agents with hepatoprotective, antioxidant, anti-inflammatory and antineoplastic activities. The experiments were carried out with outbreed and inbreed (CBA, C57Bl/6) mice. The experimental model of CCl4 hepatitis was used to estimate an antioxidant and hepatoprotective activities. The models of carrageenan- and histamine-induced mouse paw edema were applied to study anti-inflammatory effect. Using mice with transplantable tumors (Lewis lung carcinoma (LLC) and mice RLS lymphoma) it was estimated the influence of agents on tumor growth, volume areas of metastasis, dystrophic and necrotic changes in some tissues (morphometry using light microscopy). Some betulonic agents were tested under the model of experimental polychemotherapy, imitating a standard anticancer therapy ACOP (adriamycin, cyclophosfamide, oncovin, prednisolone). It is shown, that carboxymethyl and benzyl ethers of UA and it’s derivatives with acetate and glycine fragments have anti-inflammatory activity with pronounce hepatoprotective and antioxidant effects. An amides of BA possess a high anti-inflammatory and antioxidant potency. D,L-(alfa) alanine and (beta) alanine derivatives of BA and their methyl esters at single dose reduce tumor growth by 10-20% and inhibit metastasis CLL by 40-50%. Among alanine amide derivatives BA it is found an agents that have high antimetastatic activity, increase a polychemotherapy potency and decrease tissues damage.

34 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication The Study of Antioxidant Activity of Natural Organic Acids

O.N. Tchaikovskaya1,2, I.V. Sokolova1,2, L.V. Nechaev1,2, N.V. Yudina3, E.V. Mal’tseva3, E.A. Sosnin4, S.M. Avdeev4

1 Tomsk State University, 36, Pr. Lenine, Tomsk, 634050, Russia 2 Siberian Physical Technical Institute, 1, Novosobornaya Sq., Tomsk, 634050, Russia, tel: +7(3822)533426, fax: +7(3822)533034, e-mail: [email protected] 3 Institute of Petroleum Chemistry SB RAS, 3, Pr. Akademicheskii, Tomsk, 634021, Russia 4 High Current Electronics Institute SB RAS, 2/3, Pr. Akademicheskii, Tomsk, 634055, Russia

Humic substances form a dark-colored mixture, which arises from the microbial degradation of organic matter, especially dead plants. It is abundant in soil, coal and water/ Humic acids (HAs) are a major component of humic substances. It contains aromatic rings, phenolic hydroxyl and carboxyl groups, which act as binding sites for metal ions. The mechanisms responsible for the toxic effects of HAs remain unclear, although it has been reported that the damage process of HAs was accompanied by an increase of oxidative stress. Reactive oxygen species like superoxide anion radicals, hydrogen peroxide or hydroxyl radicals play an important role in pathogenesis of chronic obstructive pulmonary disease by activating signaling pathways, mediating a wide range of cellular responses. In this study, we investigated the antioxidant activity of HAs under exposure to the KrCl excilamp (the emitting maximum UV-radiation at 222 nm), the mercury lamp and Solar55 (the several emission maxima in the 320-850 nm region) lamp irradiation by voltamperemeter and fluorescence methods. HAs solutions were prepared in laboratory from dry charges. HAs from peat (T1) and Aldrich were extracted by 0.1 N NaOH solution. The HAs solutions at pH=11 were irradiated. The irradiation time was varied from 1 min to 32 min. Interestingly, when HAs solutions were irradiated with KrCl exilamp, the rate of Has phototransformation increased and the antioxidant reactivity of Has was decreased. By contrast, on exposure to radiation from Solar55 lamp the antioxidant activity was considerably increased.

The work was supported in part by Federal Target Program "Scientific and Teaching Staff of Innovative Russia" for 2009-2013 years (Projects: No. P1128 and No. 02.740.11.0444).

35 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Medications Standardization Problems

N.A. Vereshchagin

Al-Farabi Kazakh National University, Almaty E-mail: [email protected]

A variety of medications derived from medical plant raw materials (MPRW) and consisting of active ingredients has been recently appeared at Kazakhstan market. However, key properties of the plants whether poorly investigated or only one group of bioactive substances is comprehensively researched. At the same time the main problem is lack of standardization of mentioned substances. Generally, components of medical plant raw materials do not provide pharmacological effect being declared, there is no substantiation of compatibility between the components and their quantity. A problem of investigation of so-called “additives” is highly important, as medications having MPRW, especially those imported to Kazakhstan from CIS and western countries are developed non-specifically, empirically and include up to 20-30 components without adequate substantiation and control, resulting with deterioration of quality and role of an “active ingredient”. Should be mentioned that multicomponent compositions of MPRW contain much less active ingredients than healing qualitative content of a separate plant. The purpose of investigation was to optimize methods of standardization of multicomponent medications which are universal for MPRW analysis, operational technological control and finished medications control and standardization. A cumulative phytopreparation of Rosa corymbifera, manufactured by “KhimFarm” JSC was selected for investigation. In compliance to main pharmacological action requested by manufacturer, condensed hardening agents – polysaccharide and ascorbic acid were selected in order to optimize methods of standardization. The capabilities of both official pharmacopoeial methods of verification of active ingredients and methods submitted for registration for medications having similar composition have been investigated. An optimal 0.25% concentration was agreed for qualitative analysis. From our viewpoint, an optimal method for standardization of hardening agent is permanganatometry.

36 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication

The advantage of such method is a distinct color transition from dark-blue to golden- yellow, giving us easy and accurate way to define equivalence point. Certain excess rates of the results may be ignored, since phytopreparations have a very low content to be affected by concurrent oxidation under the method revealed here above. For the purposes of concentration limit studying we used seven concentrations (1%; 0,5%; 0,25%; 0,125%; 0,0625%; 0,03125%; 0,015625%). The method is functionable till 0,125% conecntration limit with further immediate noncharateristic coloration change. Alteration of pH of working solutions does not affect analysis results. Based on exprerimental data obtained, a temperature of 20-30 0C and 0,25% concentration should be optimal conditions for reaction. Weight method – polysaccharide precipitation by alcohol – was an optimal method for polysaccharide verification. The key advantages of this method – simplicity and high rate for results reproducibility. Whereas ethylene alcohol able to precipitate both polysaccharides and albumins, we have performed additional studies for albumin content, namely, ability for polysaccharides/inability for albumin to dissolve in 10% trichloracetic acid. Based on such additional study we proposed to introduce correction coefficient to analytic formula of mentioned method. Supposedly, direct titration of sodium 2,6-dichlorophenolindophenol is an optimal method to define ascorbic acid. The advantage of this method is high selectivity rate and moreover, present method is of non-indicative type making analysis much simple. A small quantity of titrant for standardization compels to utilize microburets which is the only disadvantage for proposed method, increasing human factor rate in accuracy and reproductibility. Based on experiment data obtained, an optimal temperature is room indoor temperature. The experiment has proven that pH derating increases quantity of titrant making the results certainly higher, where as pH uprating causes inverse relationship.

37 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication New in Ecdysteroids Chemistry: the Synthesis of Minor Phytoecdysteroids and Ecdysteroids Analogues

Natalya A. Veskina, Aliya Sh. Ibragimova, Ilgiz V. Galyautdinov, Victor N. Odinokov

Institute of Petrochemistry and Catalysis, Russian Academy of Sciences, 450075 Ufa, Russian Federation Fax: +007-347-2312750; e-mail: [email protected]

As it was established by us 1 [], the ecdysteroids have transformed to 9α,14α- epoxyderivatives (oxetanoecdysteroids) in alkaline metals-liquid ammonia solution in an air presence. These derivatives are interesting for the further transformations. It turned out that at the interaction of diacetonide 1 with NaBH4 the compounds 2-4 were formed, i.e. 2-4 are the products of the 6-ketogroup reduction and the oxetano-cycle rearrangement. The oxidation of the 2 gives ketone 5, the acid hydrolysis of which leads to 25-hydroxydacryhainansterone 6. The latter was before obtained by the dehydration of little-spread turkesterone [2]. The epimeric alcohols 3 and 4 were oxidized into the same ketone 7 which was earlier described by us. All compounds structures are proved with the 1D and 2D NMR experiments.

O O O O O O O O OH OH OH H OH H H H HO HO 1.NaBH , ТГФ-МЕОН O 4 O O O O 2.H О, ЕtOAc 2 OH + + O O O O H H H H H OH H H OH O OH 1 2 3 4

ПБХ, СHCl3

O OH O O OH O H OH H OH H OH HO O HO O 10% HClO4, МЕОН OH OH O HO O H H H O O O 6 5 7

1 Odinokov V.N.; Galyautdinov I.V.; Ibragimova A.Sh.; Veskina N.A.; Khalilov L.M.; Dolgushin F.M.; Starikova Z.A. Mend.Commun, 2008, 18, 291. 2 Bourne P.C.; Whiting P.; Dhadialla T.S.; Hormann R.E.; Girault J.-P.; Harmatha J.; Lafont R.; Dinan L. Journal of Insect Science, 2002, 2, 11.

38 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Resources and Biotechnology of the Ecdysteroid-Containing Plants

Svetlana O. Volodina, Ivan F. Chadin, Vladimir V. Volodin

Institute of biology of the Komi Sci Centre, Ural Division, Russian Academy of Science 28 Kommunisticheskaya Str., 167982 Syktyvkar, Russia E-mail: [email protected]

At present phytoedysteroids that are structurally relative to insect moulting hormones attract attention for the development of the new effective and safe adaptogenic medicinal preparations and nutritional supplements since these compounds do not have any toxic and negative hormonal effects on mammals. We developed the methodology of the screening of plants for the ecdysteroids presence. A number of perspective plant species with high ecdysteroids content are revealed among ferns and angiosperms from the geographically remote flora (European northeast of Russia, the Urals, the North Caucasus, the Ukraine, the Far East, etc). The composition and dynamics of ecdysteroids in a number of wild and cultivated plants of the genera of Rhaponticum (R. carthamoides, R. serratuloides), Serratula (S. coronata, S. tinktoria, S. quinquefolia, S. radiata, S. gmelini), Silene (S. tatarica, S. repens), Chenopodium (Ch. bonus-henricus) are studied. The method of the micro-clonal propagation of some ecdysteroid-containing species. The highly productive strains of the callus and suspension cultures of plant cells – ecdysteroids producers are obtained. The technology of obtaining the ecdysteroid-containing substance Serpisten from the overground part of Serratula coronata is developed. The results of preclinical studies showed the strongly pronounced anti-ischemic, hypolipidemic, anti-diabetic, anti-ray and actoprotective effects. On its basis three encapsulated forms of nutritional supplements (Kardisten with anti- ischemic and cardio-protective), Diasten with anti-diabetic and Adasten with immune- stimulating action) are developed and recommended for the use in geriatry and restorative medicine.

39 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Biological Activity of Sulfated Polysaccharides from Red Seaweed of Pacific Coast

Irina M. Yermak1, Anna O. Barabanova1, Victoria N. Davidova1, Ekaterina V. Sokolova1, Yong Hwan Kim2, Kwang Soon Shin2

1Pacific Institute of Bioorganic Chemistry, FEB RUS, Vladivostok159, 690022, Russia fax: 7(4232 314050), e-mail: [email protected] 2Department of Food Science & Biotechnology, Kyonggi University, Suwon, 442-760, Korea

Sulfated polysaccharides of red seaweeds – carrageenans – are widely utilized due to their excellent physical functional properties, such as thickening, gelling and stabilizing abilities. Carrageenans have been safely consumed as constituents of food products for many years and have pronounced properties useful in the biomedical field. Carrageenans have been isolated from several algae family Gigartinaceae and Tichocarpaceae collected from Pacific coast. Chemical structure of different type carrageenans (kappa-, lambda-, hybrid -kappa/beta-, kappa/iota- and new x-type carrageenan) was established. On the basis of the data received in experiments in vitro, the dependence of some immunomodulatory properties of the carrageenans on the type their structure is shown. All type polysaccharides have stimulated the production of the IL-6, IL-10 and TNF-α in the dose-dependent fashion in the human embryonal kidney cell. Kappa- and lambda- carrageenan possessed strong activity in the synthesis of IL-6, TNF-α and kappa/beta- carrageenan, induced synthesis of the antiinflamatory IL-10 at low concentration (10 ng/mL). Non gelling lambda–carrageenan showed high macrophage-phosphatase activity. The anticoagulant activity of carrageenans depends on the molecular weigh of polysaccharides, sugar composition, sites of sulfation and the distribution of sulfated units along the galactan chain. Iota/kappa carrageenans showed the potent anti-coagulant activity, which was extremely strong in low concentration of 100 g/mL. The protective effect of carrageenan on the damaging effect of LPS of gram-negative bacteria was also studied in vivo. Carrageenan was shown to increase mouse resistance to the toxic effect of LPS. The degree of protection depends on structural types of carrageenan, polysaccharides concentration and administration time and route. The activation of cells by carrageenan occurs through TLR4 receptors. The estimation of therapeutic action of carrageenans in complex therapy of patients by sharp intestinal infections was given. Carrageenan restored the system of the hemostatic and corrected parameters of immune system of organism in the course of treatment of patients with food toxic infection of Salmonella etiology.

40 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Synthesis of Biological Active [N-(2-Ethoxyethyl)piperidyl-4]propargyl Derivatives of Natural Alkaloids and Their Synthetic Analogs

Valentina K. Yu, Kaldybay D. Praliyev, Aissulu Zh. Kabdraissova, Kanitar Kanitar

Institute of Chemical Sciences, 106, Walikhanov str., Flmaty, Kazakstan, tel/fax: +7 727 2916069, e-mail: [email protected]

About half of applied medicines are presented by natural substances, their derivatives or analogues. One of priority directions in creation of new medicines is updating natural connections, alkaloids in particular, which molecular structure promotes introduction in their structure of the various functional groups strengthening major therapeutic effect of substrate or leading to biological activity change. In the Institute of Chemical Sciences the most promising compounds-leaders had been revealed among piperidine derivatives which have the N-etoxyethylpiperidine structural fragment. In this chemical family the morpholinepropargyl derivative was synthesized which was found to be low-toxic highly effective non-narcotic analgesic with expressed anti-inflammatory, antipyretic effect. R' Due to these results it was R'' reasonable to conduct research in i), ii) C2H5 O (CH2)2 N an updating direction of [N-(2- O (H,R) C2H5 O (CH2)2 N etoxyethyl-4(OH,OR)-piperidyl- O (H,R) 4)]acetylene in Mannich reaction R= (CH2)n O (Alk, Ph); R'=Ph, C6H3(OCH3)2 using alkaloids of anabasine and ; N ; N CH3 ; N O ; N S trimetazidine as well as synthetic piperidines, morpholine,

N N-CH3 ; N N-CH2CH3 ; N N ; R'' = thiomorpholine, piperazines as N amine components and paraform OCH3 and aromatic aldehydes as ; N N OCH3 N aldehydic ones. To obtain final propargylamines the reaction of Mannich is used. During the aminomethylation of [N-(2-etoxyethyl-4(OH,OR)-piperidyl- 4)]acetylene using the paraform and cyclic amines in dioxane in the presence of CuCl at the temperature of 100-110°C and 15-20 min of refluxing (i) the additional azaheterocycle – morpholine, thiomorpholine, derivatives of piperidine and piperazine, alkaloids of anabasine and trimetazidine, possessing their own pharmacological potential, – is added into the initial molecule. It is found that mono-iodide copper is the most suitable catalyst in Mannich reaction with aromatic aldehydes. The promotion of aminomethylation process with microwave radiation on aluminium oxide with MW oven capacity of 40-75 W in the presence of fresh CuI or CuCl (ii) reduces time to 60-80 sec. Among the synthesized derivatives the analgesics, spasmolytics and substances suppressing growth of pathogenic microorganisms and mycobacterias are found.

41 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Oral Communication Isolation of Substances from Plants of Hipophae rhamnoides L. Species and Study on Their Phytochemical and Antioxidant Properties

Gaukhar S. Sapargalieva, Galiya E. Zhusupova, Maira K. Murzakhmetova, Zharylkasin A. Abilov, Zulkhair A. Mansurov

al-Farabi Kazakh National University, Karasai Batyra 95a, 050012, Almaty, Kazakhstan E-mail: [email protected]

Plant resources of Kazakhstan are rich and diverse. One of such representatives is the common sea-buckthorn (Hipophae rhamnoides L.), leaves and stems of which have been studied for the first time. Plant material was gathered in August, 2009 in the Almaty region. It was established that humidity of plant material made up 7.47 %, total ash – 4.9 %, ashes, insoluble in 10 % HCl – 0.0079 %, sulphated ashes – 17.1 %. Qualitative value of the obtained substance was used as the principal criteria while working out optimal conditions for its isolation. By varying most crucial factors optimal parameters for the substance isolation have been found as: grounded to 3 mm particle size plant material was extracted with 50 % ethanol (or 50 % acetone) in ratio of 1:8 w/v, extraction lasted for 6 hours under room temperature of 20-25 0C with double repetition of the whole process of extraction. Joint extracts were filtered and concentrated to get the substances in form of dry extracts, which then were processed by chloroform, dried up and loaded into column (2x30 cm), filled with specially developed structured nanosorbent, obtained from rice husk, being an industrial waste. The column was first washed with distilled water, later on with ethanol-water solutions of different concentrations. Obtained fractions were evaluated by one-dimensional paper chromatography using standard samples in butanol-acetic acid-water system in ratio of 40:12.5:29 (1); 4:1:5 (2) and by two-dimensional chromatography (in systems 1 and 2 and in 2 % CH3COOH) using specific developers. The presence of fructose, galactose, valine, lysine, arginine, flavonols and different forms of flavan-3-ols was indicated. Hepatocyte membranes, used to study antioxidant properties of the obtained substances, were isolated according to the common methodic. Accumulation of lipid peroxidation products (LPP) in liver microsomes was assumed by the reaction with 2-thiobarbituric acid and was calculated by the intensity of developing color using method of Ohkawa et al.1 LPP in membranes was induced by Fe2+/ascorbat system and incubation of analyzed samples under 0 37 C in medium, containing 0.85 % NaCl, 50 mM KH2PO4 and pH 7.4. Optical density was measured under the wavelength 532 nm. It was established that all objects obtained antioxidant properties, herewith antioxidant effect of the substance, isolated from the stems and obtained using extraction with acetone was the highest one.

1 Ohkawa H.; Ohishi N.; Yagi K. Anal. Biochem. 1979, 95, 351-358.

42 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation

Poster Presentations Synthesis and Biological Activity of Sesquiterpene Lactone Modified by Natural Alkaloids

S.G. Klochkov, S.V. Afanasyeva, A.B. Ermatova

Institute of Physiologically Active Compounds of Russian Academy of Sciences, Chernogolovka, Russia Fax: (496)524-2525; e-mail: [email protected]

Natural sesquiterpene lactones and alkaloids have shown the wide spectra of physiological activities [1]. The lactone molecules isolated from the various plants of Asteracea family contain α-methylene-γ-lactone fragment. This fragment easily undergoes the reaction of Michael-type with the nucleophilic reagents. Sesquiterpene lactones isolated from the Inula helenium and products of their acidic transformation [2] were taken for the modification. We have used as N-nucleophiles natural alkaloids of three types: the phenetylamine derivatives (tyramine, ephedrine); the piperidine derivatives (anabasine, salsoline, salsolidine, cytisine); the indole derivatives (tryptamine, serotonine, mexamine, melatonine). The all reactions have been carried out in a mild condition with a high regio- and stereoselectivity. Using the method described above, a large number of the new compounds containing the sesquiterpene lactone fragment together with the alkaloid fragment were synthesized with high yields, e.g. the adducts of isoalantolactone with tryptamine and melatonine:

NH2 MeOH O X O + 20oC N H

X X O O O O Ac2O, Py

H 0oC N N Ac N N H H (80%) (75%) Various types of biological activities were investigated for the obtained derivatives. The perspective of studied compounds as potential effective antineoplastic drugs was demonstrated.

1 B.M.Fraga. Nat. Prod. Rep., 2006, 23, 943.

2 S.G.Klochkov, S.V.Afanaseva, A.N.Pushin. Chem. Nat. Comp., 2006, 42, 400

43 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Chemical Constituents of Leonurus turkestanicus

I.M. Isaev, M.A. Agzamova and M.I. Isaev

S.Yu.Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax (99871) 120 64 75, e-mail: [email protected]

Leonurus turkestanicus V.Krecz. et Kuprian (Labiatae) grows in Uzbekistan and it is widely applied instead of an official herb Leonurus cardiaca L. However, there are no data on a chemical content of secondary metabolites of this plant in general and about the chemical nature of the active principle in particular. Owing to it we have investigated the chemical constituents of the secondary metabolites of the grasses Leonurus turkestanicus. One flavonoid and two glycosides of the iridoid type were isolated from the methanolic extract of the plant by using a column chromatography. Flavonoid was identified with genkvanin (1). Iridoidic glycosides were determined as 8-acetylharpagide (2) and harpagide (3). The identification of compounds 1-3 carried out by using the modern methods of 1D and 2D NMR spectroscopy: 1H, 13C, DEPT, 1H-1H COSY, HSQC, IR spectroscopy.

O HO OH OH CH3O OH HO

O O OH O H H AcO H HO H O O 1 HOCH2 HOCH2 O O OH OH HO HO OH 2 OH 3

44 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Anticancerogenic Activity of Alkaloids from Haplophyllum perforatum

V.I. Akhmedjanova a, L. Angenot b, R.Sh. Shakirov a

a S. Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, M. Ulugbek Str., 77, 100170 Tashkent, Republic of Uzbekistan Fax: (998 71) 120 64 75 E-mail: [email protected], [email protected] b University of Liege, CHU, B36, B-4000 Liege, Belgium

The plant Haplophyllum perforatum Kar. et Kir. (Rutaceae), widely distributed in Uzbekistan, was used in folk medicine as sedative, analgesic, antitumoral and other drugs. It was found to produce alkaloids, lignans, flavonoids, coumarines, etc. We evaluated for the first time antitumor activities of known quinoline alkaloids glycoperine (1), graveoline (2), dictamnine (3), dubinidine (4), N-methyl-2-phenylquinolin-4- one (5), perfamine (6), perforine (7), skimmianine (8), flindersine (9), foliosidine (10), haplamine (11), haplopine (12), and evoxine (13) which were isolated from H. perforatum. Compounds were dissolved in DMSO and six different concentrations from 1 to 100 ?M were tested in vitro against two human cancer cell lines HeLa and HCT-116. The cytotoxicity was evaluated by WST-1 reagent and reading at ELISA (450 nm). Colchicine (14) was used as a positive control.

Five alkaloids proved to be active against HeLa cells with IC50, ?M: 65.0 (3), 11.6 (8), 49.4 (9), 36.5 (11), 29.4 (12), 1.1 (14). They belong to furoquinoline (3, 8, 12) and pyranoquinolin-2-one (9, 11) subgroups. Only alkaloids 3 and 11 showed moderate activity against HCT-116 cells with IC50 < 100 ?M (85.0 and 64.5, respectively). Other alkaloids 1, 2,

4, 5, 6, 7, 10, and 13 were low cytotoxical against both cancer cell lines with IC50 > 100 ?M. OCH 3 O R 1

R1 N O R N O 2 H

9: R =H 3: R1=R2=H 1 11: R =OCH 8: R1=R2=OCH3 1 3 12: R1=OH, R2=OCH3

45 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Quantification of Ecdysteroids in Insect by HPLC-MS

Alexander A. Alekseev1, Vladimir G. Vasiljev2

1 Institute of Chemical Kinetics and Combustion, SB RAS, Institutskaya str., 3, 630090 Novosibirsk, Russia. Fax: +7(383)3309150 Tel: +7(383)3333499 E-mail: [email protected] 2 Novosibirsk Institute of Organic Chemistry, SB RAS, Lavrentyev ave., 9, Novosibirsk 630090, Russia Fax: +7(383)3309752 Tel: +7(383)3307864 E-mail: [email protected]

Ecdysteroids are one of the main insect hormones, which involve in regulation of the important physiological processes sach as molting, metamorphosis, reproduction and insect stress-reaction. We developed the simple, fast and sensitive method for sample preparation and quantification of ecdysone and 20-hydroxyecdysone in insect hemolymphe and whole body homogenates by RP-HPLC coupled with a mass spectrometr (LC–MS). The procedure of sample preparation includes extraction by MeOH, delipidization, concentration by solid-phase extraction with Oasis HLB/Diapak C16 cartridge. LC–MS analyses were carried out on a Agilent 1200 LC system with a Hybrid Quadrupole Time of Flight mass spectrometer (microTOF-Q, Bruker) equipped with Atmospheric Pressure Chemical Ionization (APCI). The analytes were separated on a RP-C18 column. The mobile phase was acetonitrile: water (15:85, v/v). A 25-S-inokosterone was used as internal standard. The limits of quantification for 20-hydroxyecdysone and 25-S-inokosterone (internal standard) were 5 ng/mL, for ecdysone was 10 ng/mL. The method was succesfully applied to analysis of ecdysteroids in imago Drosophila whole body extracts and Galleria hemolymphe.

This study was supported by grant of Russian Foundation for Basic Research #10-04- 01462-a; by Integration Grant #27 from Siberian Division of Russian Academy of Sciences and Chinese Academy of Sciences

46 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Chemical Composition of the Essential Oil of Thymus punctulosus

L.I. Alekseeva 1, A.G. Bystrushkin 2, I.V. Gruzdev 1

1 Institute of Biology, Komi Research Center, Ural Division, Russian Academy of Sciences, Syktyvkar, Russia e-mail: [email protected] 2 Botanic garden, Ural Division, Russian Academy of Sciences, Ekaterinburg, Russia

The genus Thymus (Lamiaceae) is popular in traditional medicine of many countries and peoples as a valuable medicinal raw materials. Infusions of thyme are encouraged to take as an expectorant and analgesic for acute and chronic bronchitis. For members of the genus Thymus shown antimicrobial, antioxidant and spasmolytic activity. The long-term studies chemical composition thyme summarized in the review «Thyme: The genus Thymus». Of great interest are studies thyme with small ranges, and in particular, endemic. Thymus punctulosus – petrofit, mountain-steppe and mountain-steppe endemic of the South Urals and the southern part of the Middle Urals. T. punctulosus is half-shrub with one or more several long-standing stalks and branching rising axial shoots. Vegetative not breed, although contact with the soil of the plant can form adventitious roots. Chemical composition of T. punctulosus has not been previously studied. Sampling of plant material was carried in natural populations of T. punctulosus in early July 2008. Populations of I is located in the vicinity of Troitsk, Chelyabinsk Region (South Ural) on outcrops of limestone on the River Uy. Populations of II is located in the vicinity of Kizilsk, Chelyabinsk Region (South Ural) on outcrops of limestone on the River Ural. According to chromatography-mass spectrometry the major components of essential oils of T. punctulosus for the population I are trans-nerolidol (24,83%) and caryophyllene oxide (19,04%), for population II trans-nerolidol (61,39%). Essential oils of T. punctulosus contain mostly sesquiterpenes – 71,88 and 82.97% for the populations of I and II, respectively. In the essential oils of T. punctulosus of the population of I the highest content of sesquiterpenes are for bicyclic sesquiterpenes – 38,57% and acyclic sesquiterpenes – 23,96%, in the population of II – for acyclic sesquiterpenes – 59,51%. Differences in chemical composition of essential oils of T. punctulosus from different populations may be caused by both genetic and environmental factors, as previously noted for Thymus spinulosus.

47 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Improving Effects of Semen Flax Extract on Hemorheological Abnormalities in Ovariectomized Rats

Anna M. Anischenko1, Tatyana M. Plotnikova2, Natalia V. Vasil’eva2

1 Institute of Pharmacology, Siberian Department, Russian Academy of Medical Sciences, 2 Siberian State Medical University, 634050, Moskovskii tract 2, Tomsk, Russia Fax: (83822) 41-42-35 E-mail: [email protected]

Impaired hemorheology has been demonstrated in post-menopausal women is risk factor of cardiovascular complication of the climacteric syndrome. High content of lignans (phytoestrogens) justifies research of its influence on blood viscosity, deformability and acid resistance of erythrocytes in ovariectomized rats. We studied three groups of female Wistar rats with weight 300–350 g divided on 3 groups: group 1 – sham-operated rats, group 2 – ovariectomized rats, group 3 – ovariectomized rats, receiving p.o. flax semen extract (40 mg/kg daily for 14 days). Content of secoisolariciresinol in flax semen dry extract is 42% (HPLC). Ovariectomy in rats (group 2) increased the whole blood viscosity as a result of erythrocyte deformability impairment. Resistance of erythrocytes for acid lysis was decreased too. Treatment of ovariectomized rats with flax semen extract (group 3) increased the resistance of erythrocyte membrane for acid lysis (by 9%) and improved the erythrocyte deformability (by 19–29% at shear rates 90–890 s–1). In consequence of these effects, whole blood viscosity decreased by 10–13% at low shear rates and by 4–6% at high shear rates. Thus, course administration flax semen extract improves the hemorheological parameters in ovariectomized rats. These results confirm that flax semen extract has considerable potential as substantial component for the development of new drug in improving of hypoestrogenemia-induced hemorheological abnormalities.

48 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis of C-20 Pyridine Analogs of Lupane-Type Triterpenoids

Anna N. Antimonova, Natalya I. Petrenko, Elvira E. Shults, Genrikh A. Tolstikov

Novosibirsk Institute of Organic Chemistry, SB RAS, Novosibirsk, Russia, e-mail [email protected]

Structural modifications of lupine-type triterpenes is the perspective route to the construction of new highly potent antitumor and anti-HIV agents [1]. The target of this work is synthesis of novel compounds with change of isopropenyl substituent to pyridine ring. So, by the reaction of betulin diacetate (1) with acetyl chloride in the presence of ZnCl2 and treatment of the formed pyrilium salt with ammonium 2,6-dimethyl-20,29,30-tris-nor- pyridyllupane (2) was isolated in the yield of about 35%. The same compound was formed by using betulin (3) as the starting compound. The reaction also proceeds with corresponding 3- keto-derivative (4) with the formation of heterocyclic compound (5). The influence of the reaction conditions, the structure of lupine triterpene and the nature of the Lewis acid will be discussed.

29 20 O 30 H AcCl, ZnCl2 3 R - CH2OAc ZnCl3 R2 2 R1 R R1

1 2 3 1. R = OAc, R = H, R = CH2OAc 1 2 3 3. R =OH, R = H, R = CH2OH 1 2 3 NH3 4. R + R = O, R = CH2OH

N H

CH2OAc R2 R1 2. R1= OAc, R2= H, 5. R1+ R2= O

This work was supported by the Russian Foundation for Basic Research (project no. 09-03- 00183).

1 Kuo, R.-Y.; Qian, K.; Morris-Natschke, S.-L.; Lee, K.-H. Nat. Prod. Rep., 2009, 26, 1321.

49 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis of Six 3-Methyl-6-(1-methylethenyl)cyclohex-3-en-1,2- diol Stereoisomers with High Optical Purity

O.V. Ardashov, I.V. Il’ina, K.P. Volcho, N.F. Salakhutdinov

N.N. Vorozhtsov Novosibirsk Institute of Organic chemistry SB RAS 630090, Lavrentiev av., 9, Novosibirsk, Russia e-mail: [email protected]

Recently it was found that (1R,2R,6S)-3-methyl-6-(1-methylethenyl)cyclohex-3-en-1,2- diol ((1R,2R,6S)-1) with 60% enantiomeric excess possess high anticonvulsant activity [1]. It is known that the biological activity of the terpenoids is depend sufficiently upon their absolute configuration. Nevertheless biological activity of the isomers of compound 1 isn’t investigated still. In this work we successfully synthesized six of all eight stereoisomers of compound 1 with high optical purity starting from the widespread monoterpenes (+)- and (-)-pinene in accordance with the scheme.

O OH O a, b, c, d, e f g OH OH OH (+)- -pinene (1S,2S,6R)-1 ee 97% ee 97% O OH OH a, h, b, i f e OH OH OH

(1R,2S,6R)-1 ee 97% (1S,2R,6R)-1 ee 97%

OH OH OH a, h, b, i, f a, b, c, d, e, f g, e OH OH OH (-)- -pinene (1S,2R,6S)-1 ee 94% ee 94% (1R,2R,6S)-1 ee 94% (1R,2S,6S)-1 ee 94%

a: Pb(OAc)4; b: KOH/MeOH; c: Na2Cr2O7; d: H2O2/NaOH; e: LiAlH4; f: K10

clay; g: DMSO/(COCl)2, h: AcOH; i: t-BuOOH/VO(acac)2.

The authors are grateful to the Russian Foundation for Basic Research for financial support (grant N 08-03-13516).

1 Il’ina I.V., Bolkunov A.V., Dolgikh M.P., Volcho K.P., Tolstikova T.G., Salakhutdinov N.F. Patent RU 2355390, 2009.

50 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Gas-Chromatographic Analysis of the Volatile Compounds of Halostachys belangeriana

1 2 2 1 G.Zh. Baisalova , G. Ozek , K.H.C. Baser , S.B. Rakhmadieva

1 Department of Chemistry, Faculty of Natural sciences, Eurasian national university, 010008, Astana, Kazakhstan 2 Department of Pharmacognosy, Faculty of Pharmacy, Anadolu University, 26470, Eskisehir, Turkey, e-mail: [email protected]

Halostachys (Chenopodiaceae) is represented by only one species in wet saline soils, sea shores and by salty lakes cores in Kazakhstan.1 H. Belangeriana was collected in Kzylorda region in the South Kazakhstan.The volatiles were obtained from the crushed over ground parts of Halostachys Belangeriana by hydrodistillation in Clevenger type apparatus. The volatiles were analyzed by capillary Gas Chromatography-Flame Ionization detection (GC/FID) and Gas Chromatography–Mass spectrometry (GC/MS) using an Agilent 5975 GC-MSD system. The same column and analysis conditions were used for both GC/MS and GC/FID. HP-Innowax FSC column was used with helium as a carrier gas. The GC-FID analysis was carried out using an Agilent 6890N GC system. In order to obtain same elution order with GC/MS, simultaneous injection was done by using the same column and appropriate operational conditions. Identification of the volatile constituents was achieved by parallel comparison of their retention indices and mass spectra with data stored in the Wiley GC/MS Library, MassFinder software 3.0, Adams Library, NIST Library and Baser Library of Essential Oil Constituents. Relative percentage amounts of the separated compounds were calculated from FID chromato-grams. Twentyseven compounds were identified. The major volatile components were found as hexadecanoic acid (24,3%), hexahydrofarnesyl acetone (14,5%), tricosane (7,3%), pentacosane (7,1%), heptacosane (5,1%), heneicosane (2,7%), 4-vinyl guaiacol (2,6%), 3,4- dimethyl-5-pentylidene-2(5H)-fura-none (2,2%), docosane (2,2%).

1 Baitenov, M. In Flora of Kazakhstan, Nauka; Almaty, 1999; pp 272-273.

51 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Hypericum perforatum L. as Potential Antidiabetic Medicinal Herb

Irina I. Bayandina, Tatyana P. Kukina

Novosibirsk State Agrarian University Dobrolubova st., 160, 630039, Novosibirsk, Russian Federation E-mail: [email protected] Novosibirsk Institute of Organic Chemistry, Siberian Branch of the Russian Academy of Sciences 9 prosp. Acad. Lavrent’ev, 630090 Novosibirsk, Russian Federation E-mail: [email protected]

The plant Hypericum perforatum L. is used in herbal medicine externally for the treatment of skin wounds, eczema and burns and internally for diseases of the alimentary tract. Many studies have shown that Hypericum perforatum L. extract (HPE) has antiviral activity including anti-HIV and antiretroviral action, antidepressant-like and other CNS- modulating effects. The literature data showed the effects of a standardized HPE in rats with experimentally induced hyperglycemia. Results of oral glucose tolerance test were performed to evaluate the effects of the extract on peripheral glucose utilization in normal rats. These results indicate that stimulation of insulin release could be involved in the observed effects of the extract in the type-2 diabetes model used. Standardized HPE a beneficial effect in a rat model and preliminary observations indicate that hyperforin may be at least partially responsible for the observed activity though additional evidence will be provided by future studies1. The action of summary extract may be connected with high content bio-active constituents such as quercetin, rutin and other flavonoids. These compounds are known as strong α-glucosidase inhibitors, decreasing blood glucose level. Some researches connect α- glucosidase inhibition activity with content of phenolics. Several other therapeutically interesting bio-active phenolic components of the herb are also known due to strong antioxidant properties. Our investigation by mass-spectrometry chromatographic analysis of hexane and methyl- tert-butylic ether extracts of St. Johns wort Hypericum perforatum L. led to discovery of new acid and neutral components: aliphatic acids (C21 – C34), 3- and 9-hydroxyacids (C12 – C18), dehydroabietic acid, gentriakontan, methyl tetracosanol, pentacosanol, β-amirine, methyl oleanolate, campesterol, stigmasterol, squalen. Predominant triterpenes of St. Johns worth are β-sitosterol, β-amirine and methyl oleanolate. Polyprenols were isolated and identified in free and acetate forms2. All those compounds take part on summary biological activity including α-glucosidase inhibition.

1 Husain, G.M.; Singh, P.N.; Kumar V. Drug Discov Ther. 2009, 3, 215-220. 2 Kukina, T.P.; Bayandina, I.I.; Pokrovsky, L. M. Chemistry of plant raw material, 2007, 3, 39-45.

52 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation The Synthesis of Hybrid Structures Consisted of Sesquiterpene Methylene Lactone and Alkaloid

Alexander V. Belovodskii1, Elvira E. Shults1, Khusnutdin M. Shakhidoyatov2, Genrikh A. Tolstikov

1 Vorozhtsov Institute of Organic Chemistry Siberian Branch, Russian Academy of Sciences, Lavrentjev Ave. 9, 630090, Novosibirsk, Russia Fax: (383)330-85-33, E-mail: [email protected] 2 Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences of the Republic of Uzbekistan, M. Ulugbek Ave. 77, 100170, Tashkent, Uzbekistan

Sesquiterpene lactones are the characteristic secondary metabolites of plants Asteraceae family. These compounds attract attention of many scientists at least last 50 years caused by diverse spectrum of biological activities including anticancer, anti-inflammatory and antiviral properties. Molecules bearing the α-methylene-γ-lactone motif have a special significance within such compounds. The new method for transformation structures mentioned above by introduction of an aromatic substituent was recently offered by authors1; available metabolites of Inula helenium2 including isoalantolactone 1 were used as example. Now we report the cross-coupling reaction of isoalantolactone 1 and haloderivatives of two different type alkaloids: 5-iodolappaconitine 2 and 6-bromodeoxyvasicinone 3. As outlined in scheme the reaction proceeds smoothly in “standard” Heck conditions and gives corresponding products 4-8 in yields from moderate to high. The quite complicate structure of lappaconitine's aconitane residue hampered the separation stage and decrease yield of product.

Thus, the way for preparing novel potentially biological active hybrid structures is proposed. Comparison the data obtained and ours earlier observations1 reveals that there is no marked difference with transfer from simple arylhalides to complicate alkaloid structure.

This work was supported by the Russian Foundation for Basic Research (project no. 08-03-00340).

1. Belovodskii, A.V.; Shul’ts, E.E.; Shakirov, M.M.; Tolstikov, G.A. Doklady Chemistry 2009, 426, 138-142. 2. Milman, I.A. Chemistry of Natural Compounds 1990, 26, 251-262.

53 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation On Phytoadaptive Activity of Lonicera caerulea L.

Irina G. Boyarskikh1 and Larisa A. Yevtukhova2

1 Central Sibirian Botanical Garden. Zolotodolinskaya St. 101, Novosibirsk, Russia. email: [email protected]. 2 The F Ssorina Gomel State University

Honeysuckle (Lonicera L.) belongs to the family Caprifoliaceae Juss. The genus contains more than 200 species; only representatives of the subsection Caeruleae Rehd. distributed in the temperate zone of Eurasia and North America have edible fruits. Medicinal properties of fruits are determined by presence of biologically active substances. Honeysuckle is a valuable source of vitamin C (40-90 mg/100 g), in addition, vitamin K (0.06-0.10 mg/100 g) and carotenoids (0.25-0.52 mg/100 g). Sum of sugars in fruits is 1.6-1.8 % on the basis of raw stuff, including glucose – 54 %, fructose – 24 %, sorbitol – 15 % and inositol – 4 %. Fruits are a valuable source of bioflavonoids – up to 1800 mg/100 g. These substances are represented by flavones (catechins, proanthocyans), oxycinnamicacids and flavonols. Biologically active iridoic glycosides [1] were found in honeysuckle. Fruits possess high antioxidant activity which depends directly on the content of phenol compounds [2]. In experiments on mice honeysuckle extracts with a high content of phenol compounds increased production of prostaglandin playing the role of immunomodulators. To reveal phytoadaptive activity of Lonicera caerulea L. of Altaian origin, influence of biologically active substances in fruits on removal of radioactive Cs from rats was studied. Patterns of accumulation and removal of Cs-137 depending on a kind of plant sorbent were obtained in the experiment. The analysis of the results showed that use of honeysuckle fruits in feeding animals increased removal of Cs-137 by 15 % in comparison with the control. The chemical analysis of raw material for content of pectic substances and correlation with the results of the experiment showed that a quantity of pectic substances contained in plant samples was not a primary reason for increase of removal rate of radioactive Cs from animal organisms. In this regard it is suggested that a complex of biologically active substances of L. caerulea has a radioprotective effect.

1 Streltsyna S.A., Sorokin A.A., Plekhanova M.N. et al. Agrarian Russia. 2006, 6, 67-72 2 Thomson M.M., Chaovanalikit A. Acta Hort.(ISHS). 2003. V. 626, 65-72. 3 Jin X.H., Ohgami K., Shiratori K., et al. Exp. Eye Res. 2006. V.82, Issue 5, 860-867.

54 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Enzymatic Hydrolysis of Miscanthus Conversion Products

Vera V. Budaeva

Institute for Problems of Chemical & Energetic Technologies, Siberian Branch, Russian Academy of Sciences, ul. Socialisticheskaya, 1, 659322 Biysk, Altai krai, Russia Fax: 8(3854)30-17-25 E-mail: [email protected], [email protected]

In elaboration of a progress in research and practical realization of non-wood feedstock conversion (specifically Miscanthus) achieved abroad [1, 2], this paper is the first to report results of study on enzymatic hydrolysis using a Russian enzyme complex of products from chemical and thermobaric treatments of Miscunthus sinensis grown on plantations of the Institute of Cytology and Genetics SD RAS in 2008. Soluble carbohydrates in hydrolyzates of pretreated Miscunthus are a substrate for ethanol nd a butanol in the chemical and biotechnological procedure of Miscunthus conversion which is being developed at IPCET SB RAS.

1 M.B. Jones, M. Walsh. Miscanthus: For Energy and Fibre. Publisher: Earthscan, 2001, 192. 2 Hannah K. Murnen et al. Optiization of ammonia fiber expansion (AFEX) pretreatment and enzymatic hydrolysis of Miscanthus giganteus to fermentable sugars, Biotechnol. Prog., 2007, 23, 846-850.

55 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis and Antioxidant Activity of Alkylthiomethyl Derivates of Quercetin and Dihydroquercetin

Ivan M. Bugaev, Aleksandr E. Prosenko

Research Institute of Antioxidant Chemistry, Novosibirsk State Pedagogical University, 28 ul. Vilyuiskaya, 630126 Novosibirsk, Russian Federation Fax: +7 (383) 244 1856 E-mail: [email protected]

Quercetin and dihydroquercetin are effective natural antioxidants and are widely used in the compositions of dietary supplements in recent years. The low lipophilicity of the flavonoids may reduce their bioavailability, and also makes them difficult to use for the stabilization of fats and oils. To solve this problem requires modification of quercetin and dihydroquercetin by lipophilic groups. Research conducted at Research Institute of Antioxidant Chemistry show that antioxidant activity of phenols sharply increases at transition to them alkylthiomethyl derivatives. We have assumed that introduction of dodecylthiomethyl groups in molecules of quercetin and dihydroquercetin will solve a problem with solubility, and also will increase antioxidant activity. Attempts of synthesis of target compounds by standard methods was failed. During the further work we had been discovered new reaction on which basis the effective method for thiomethylation of phenolic compounds has been developed. By this method we were able to prepare dodecylthiomethyl derivatives of quercetin and dihydroquercetin. As we expected, solubility in fats and antioxidant activity of modified flavonoids have essentially increased.

56 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Obtaining Mannanooligosaccharide Additive by Means of the Mechanoenzymatic Hydrolysis of Yeast Biomass

A.L. Bychkov1,3, E.I. Ryabchikova2, K.G. Korolev1,3, O.I. Lomovsky1

1Institute of Solid State Chemistry and Mechanochemistry, Kutateladze 18, Novosibirsk, 630128, Russia, 2Institute of chemical biology and fundamental medicine, Lavrentiev Ave.8, Novosibirsk, 630090, Russia, 3Research and Education Centre “Molecular Design and Ecologically Safe Technologies”, Novosibirsk State University, Pirogova 2, Novosibirsk, 630090, Russia Fax: (383) 332-28-47 E-mail: [email protected]

The obtaining of mannanooligosaccharide additive1 by means of the mechanoenzymatic hydrolysis of yeast biomass is studied by methods of chemical analysis and electron transmission microscopy. To study the structure imperfection of cell wall glycoproteins the treatment by osmic acid and ammonia complexes of silver nitrate was offered. Reactions of proteins with osmic acid allow to visualize the accessible proteins, and the reaction of carbohydrates with silver ammonia complexes allow to reveal the defects in polysaccharide components of cell wall. To measure he t yield of mannanooligosacharide the chemical analysis was offered. After the mechanic activation most of the cells are disintegrated. Interaction of biomass with silver complexes leads to silver nanoparticles generation. Evidently, the disordering of the glucan layer occurs and elemental silver is formed on the defects, accessible for reagents. After the enzymatic treatment of activated yeast biomass the increase of number of electron impermeable nanoparticles both in external and internal mannanoprotein layers of the cell wall is shown on TEM images. And even higher concentration of silver nanoparticles in glucan layer is observed. That assumes both the imperfection of glucan and the accessibility of cell wall proteins increase after mechanical activation and further enzymatic hydrolysis of glucan. Chemical analysis proves the preliminary mechanical activation substantially influences the degree of conversion of yeast cell wall during the further enzymatic hydrolysis. The yield of mannanooligosacharide is increased from 1.3 to 3.8 %.

The research was done with the support of CRDF grant № RUX0-008-NO-06/BP4M08.

1 A.E. Wold, J. Mestecky, M. Tomana, et al. Infection and Immunity. 1990. № 58. V. 9. p. 3073-3077.

57 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Research of Siberian Fir Phenol Compounds Using Chromatographic Profiling Method

E.I. Chernyak, A.I. Vyalkov, S.V. Morozov

Novosibirsk Institute of Organic Chemistry, SB RAS Lavrentyeva ave, 9, 630090, Novosibirsk, Russia Fax: 8(383)330-97-52 E-mail: [email protected]

Siberian fir Abies Sibirica woody greenery is the rich source of wide action spectrum biologically active substances, based on which the preparations used in medicine, agriculture, food and cosmetic production re a received. All natural complexes extracted are multicomponent mixtures of organic compounds. Studying their structure demands chemical and physical research complex application. Today analyzing multicomponent mixtures of biologically active substances extracted from renewable raw materials, the method based on receiving chromatographic profiles (“fingerprints”) and stating individual-group composition of complicated objects of nature is used. Chromatographic profiles of Siberian fir woody greenery phenol complex were received by methods of chromato-mass-spectroscopy and high pressure liquid chromatography. Using “ionic extraction” method based on molecular and characteristic fragmentary ions and diagnostic UV-spectral relations data, chromatographic profiles of main Siberian fir woody greenery phenol compounds – hydroxybenzoic and hydroxycinnamic acids, flavonoids, lignans (pic.1) were obtained. Quercetin, kaempferol and isorhamnetin (flavonoids); 3,4-divanilyltetrahydrofuran, lariciresinol, isolariciresinol, matairesinol, piniresinol, hydroxymatairesinol (lignans); 4- hydroxy-, 4-methoxy-, 4-hydroxy-3-methoxy-, 3,4-dimethoxy-, 3,4,5-trimethoxycinnamic acids; 4-hydroxy-, 4-methoxy-, 4-hydroxy-3-methoxy- and 3,4,5-trimethoxybenzoic acids were identified when analyzing spectral-chromatographic profiles.

58 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Steam-distillation and Simultaneous Hydrodistillation – Solvent Extraction in Essential Oil Preparation from Fresh Plant Material

Dmitry V. Domrachev, Alfred L. Revutsky, Alexey V. Tkachev

Novosibirsk Institute of Organic Chemistry, Lavrentjev Ave. 9, 630090, Novosibirsk, Russia Fax: +007 (383) 330-97-52 E-mail: [email protected]

Two mobile devices for essential oil preparation by steam-distillation (SD) and simultaneous hydrodistillation – solvent extraction (SHD/SE) have been developed and designed for field research to prepare volatiles from fresh plant material. Both devices were made of stainless steel and tested in expeditions to the high mountain region of Altai. Samples of essential oils prepared from fresh plants by SD and SHD/SE were studied by GC-MS using a quadruple MS (Hewlett-Packard MSD 5971) coupled to a HP 5890/II GC fitted with an HP-5ms capillary column. Compositions of plant volatiles from Juniperus sabina L. and Picea obovata Ledeb., prepared by SD and SHD/SE are given in the Table. Both devices provide one and the same set of compounds, but the percentage of the volatiles prepared by SD and SHD/SE are not the same. The main difference is in ratio of volatile and semi-volatile compounds, and hydrocarbons and oxygen-containing derivatives, the content of semi-volatiles and oxygenated derivatives being greater when SHD/SE is used.

Table. Main constituents (%) of essential oils from Juniperus sabina and Picea obovata retention P. obovata J. sabina indices compound SD SHD/SE SD SHD/SE 932 α-pinene 5.49 3.63 58.89 48.85 947 camphene 10.46 7.74 0.39 0.35 975 β-pinene 1.04 1.20 2.35 2.24 991 β-myrcene 2.15 2.21 2.37 2.31 1010 3-carene 2.61 3.54 1.21 0.89 1028 limonene + β-phellandrene 6.76 6.20 2.07 1.82 1031 1,8-cineole 0.28 1.76 – – 1144 camphor 0.18 3.53 – – 1148 camphene hydrate 0.40 4.99 – + 1166 borneol 0.28 2.40 – + 1191 α-terpineole 0.12 1.56 0.03 0.38 1287 bornyl acetate 57.92 45.01 0.95 1.34 1517 γ-cadinene 0.18 0.05 1.84 0.92 1527 δ-cadinene 0.99 0.25 5.79 5.25 1553 elemol – – 1.74 5.83 1578 germacrene D-4-ol – – 2.28 1.97 1643 T-cadinol + T-muurolol 0.58 0.65 2.54 3.64 1658 α-cadinol 0,78 1,13 3,02 5,69 2058 isocembrol 0,27 2,34 – –

59 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Enantiomeric Composition of Monoterpene Hydrocarbons Isolated from Siberian Conifers

Dmitry V. Domracheva, Elena V. Karpovaa,b, Alexey V. Tkacheva,b

a Novosibirsk Institute of Organic Chemistry, 630090, Novosibirsk, Russia b Department of Natural Sciences, Novosibirsk State University, Pirogova str., 2, 630090, Novosibirsk, Russia Fax: +7 (383) 330-97-52 E-mail: [email protected]

Enantiomeric composition of monoterpene hydrocarbons from Pinus sibirica, Pinus silvestris, Picea obovata, Larix sibirica, Abies sibirica, Juniperus sabina, Juniperus pseudosabina was studied. Essential oil samples were obtained by steam distillation from freshly collected plant material in Tomsk region and north-east Kazakhstan. Fractions of monoterpenes were prepared by vacuum distillation of the crude oils. Enantiomeric compositions were determined by chiral GC-MS using an Agilent 6890/5973N EI/PCI instrument equipped with a CycloSil-B chiral capillary column. Ratios of enantiomers of monoterpenoids are given in the Table.

Table. Enantiomers ratio (–):(+) of monoterpene hydrocarbons from Siberian conifers Abbreviations: Psib – Pinus sibirica, Psilv – Pinus silvestris, Jsab – Juniperus sabina, Jpseudosab – Juniperus pseudosabina Psib1 Psib2 Psilv1 Psilv2 Psilv3 Jsab Jpseudosab α-pinene 8:92 8:92 26:74 31:69 23:77 92:8 91:9 camphene 28:72 27:73 84:16 89:11 82:18 >99:1 >99:1 β-pinene 14:86 15:85 11:89 10:90 11:89 ~1:8 ~1:8 limonene 94:6 94:6 94:6 77:23 87:13 92:8 95:5 β-phellandrene >99:1 >99:1 >99:1 >99:1 >99:1 22:78 29:71

Abbreviations: Pic – Picea obovata, Lar – Larix sibirica, Ab – Abies sibirica Pic1 Pic2 Pic3 Pic4 Lar1 Lar2 Lar3 Lar4 Ab1 Ab2 Ab3 α-pinene 90:10 84:16 81:19 83:17 78:22 95:5 93:7 78:22 53:47 41:59 45:55 camphene 88:12 83:17 89:11 83:17 89:11 96:4 93:7 98:2 96:4 95:5 96:4 β-pinene 2:98 7:93 12:88 7:93 3:97 4:96 4:96 3:97 14:86 9:91 4:96 limonene 98:2 81:19 91:9 88:12 79:21 70:30 67:33 76:24 79:21 81:19 89:11

This work was supported in part by Russian Foundation For Basic research (Grant # 07-03-00620-a).

60 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Chromatographic Isolation and Hightroughput Screening of Potencial Cytostatic on the Base of Extracts from Asteraceae Plants

A.B. Ermatova, A.V. Chudinov, O.V. Novikova, I.M. Bravova, S.G. Klochkov

Institute of Physiologically Active Compounds of Russian Academy of Sciences, Chernogolovka, Russia Fax: (496)524-2525; e-mail: [email protected]

The plants of Asteraceae family are actively applied in national medicine for treatment of a wide spectrum of diseases, including oncological. The basic active substances of plants of the above family are various sesquiterpene lactones [1]. Sesquiterpene lactones may be considered now as a promising source of new drugs taking into account the possibility of easy cultivation of plants of genius Inula and high lactones content in certain plant species [2]. Use of the traditional approach of research of biologically active compounds of plants does not allow conducting wide scale researches in short time. We had been offered a way of determination of potential cytostatic activity of components of plant extracts based on semi- preparative chromatographic separation of extracts and high-throughput screening of biological activity. During chromatographic separation of extracts it is simultaneously formed two plates – deep-well plate for different types of biological screening and an analytical plate for LC-MS/MS investigations. The screening plate goes for complex testing on cytostatic, antioxidant activity, etc. The analytical plate after activity detection is investigated with the use of mass-spectrometry. In the present report the data on screening research of 80 extracts of plants of Asteraceae family on cytostatic and antioxidants activity were demonstrated. High level and frequency of cytotoxic activity of revealed sesquiterpene lactones from plants of Asteraceae family demonstrate the outlook of further use of these compounds as scaffolds within hit-to-lead program and their further mechanism based screen.

1 S.Zhang, Y.K.Won, C.N.Ong. Curr. Med. Chem. Anticancer Agents, 2005, 5, 3, 239-49

2 S.G.Klochkov, S.V.Afanaseva, N.C.Zefirov. Technol. Liv. Systems. 2008, 5, 5-6, 31-39

61 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Clerodane Diterpenoids from Pulicaria Gnaphalodes

K.A. Eshbakova

S.Yu.Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, 100170, Tashkent, Republic of Uzbekistan E-mail: [email protected]

Perennial grassy plants genus Pulicaria (Compositae family) widely distributed in the Central Asia and are the rich source of terpenoids and flavonoids. In continuation of a study of the chemical composition of aerial parts P. gnaphalodes during blossoming time, which grows in the Djizakh region of Uzbekistan. P. gnaphalodes used in traditional and non traditional medicine as antifungal activity. We have isolated by column chromatographic method two compounds, diterpenoids characters from ethylacetate fraction. Structures isolated compounds have been established on the basis of their physical and chemical properties and the analysis of their spectral data IR, UV, 1H, 13C NMR, DEPT. The isolated compounds have been identified as salvifolin (1) and salvicin (2). These compounds possess strongly pronounced antihyperglycemic and hypolipidemic activity. Salvifolin and salvicin have been isolated from P. gnaphalodes for the first time.

H OH

HOOC OH

62 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Flavonoid from Scutellaria holosericea

H.Sh. Kamoldinov, K.A. Eshbakova

S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, 100170, Tashkent, Republic of Uzbekistan E-mail: [email protected]

Genus Scutellaria L. (Lamiaceae) widely distributed in the word and are the rich source of flavonoids. In flora of Uzbekistan 32 kinds Scutellaria L is. From genus Scutellaria have been isolated in the middle of phenol compounds – chalcones, flavones, flavonoles, flavanones, isoflavones, biflavones, lignoflavones and others. Many species widely used in traditional and non traditional medicine as antibacterial preparation, anthelmintic, malaria, choleretic, hepatoprotective, detoxifying, stomatological, antioxidant properties, morbus hypertonicus, anti-inflammatory action and others. In continuation of our studies of the chemical composition of aerial parts Scutellaria holosericea during blossoming time, which grows in the Surchandarya region of Uzbekistan, we have isolated by chromatographic method compound 1 from ethylacetate fraction. Structure 1 have been established on the basis of its physical and chemical properties and the analysis of its spectral data IR, UV (with NaOAc), 1H, NMR. The isolated compound has been identified as 5-hydroxy-7-metoxyflavone (1). This compound has been isolated from S. holosericea for the first time.

CH3O O

OH O

63 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Qualitative Analysis of Coumarins in Seeds Nigella sativa L., Growing in Uzbekistan

K.A. Eshbakovaa, Sh.A. Temurovab, I.K. Azizovb, N.D. Abdullaeva

a) S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, 100170, Tashkent, Republic of Uzbekistan E-mail: [email protected] b) The Tashkent pharmaceutical institute, Tashkent, Uzbekistan

We have studied coumarins seeds Nigella sativa, growing in Uzbekistan. Qualitative definition coumarins carried out methods of a paper chromatography in system of solvents hexan – etylacetat (3:1). At survey dried up chromatogram’s in UV-light one stain with Rf 0.6, having brightly blue fluorescence, characteristic for derivatives 7- oxycumarin’s, and giving positive reaction with Pauli's reactant is revealed. At chromatograph comparison of the found out substance to authentic samples of "witnesses" it is identified roughly as umbellipherone. For detailed studying componential structure of phenolic connections used a method of highly effective liquid chromatography (HPLS). Research (USA) with the subsequent computer processing the received data (the standard program «3D Chem Station») carried out on highly effective chromatograph «Agilent 1100 series» firms «Agilent Technologies». Detecting carried out with help UV-of the detector at length of a wave of 254 nanometers on it is inverted – a phase column in the size 2,5x150mm, filled silicagel with the size of particles 5 microns with imparted oktadesil groups ODS S-18, at room temperature. As a mobile phase used a mix metanol-50 mmol/l a solution of the phosphatic buffer (pH 3), gradient changing from a parity 20 : 80 till 75:25. Speed of submission eluent’s made 0,5 ml / mines, duration of the analysis of-25 minutes. In chromatograph entered on 20 mkl researched extraction and solutions of standard samples. Identification of substances on chromatogram’s researched extraction Nigella sativa carried out by comparison of time of their keeping to from time to time keeping standard samples. Thus, as a result of the carried out researches it has been revealed, that in seeds Nigella sativa contain such coumarins like: umbellipherone and bergapten. With help HPLS it was possible not only to confirm qualitative structure of the phenolic connections established earlier by a method of a paper chromatography, but also considerably to expand it.

64 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Flavonoid from Roots Scutellaria schachristanica

Z.O. Tashmatov, K.A. Eshbakova, Kh.M. Bobakulov, N.D. Abdullaev

S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, 100170, Tashkent, Republic of Uzbekistan E-mail: [email protected]

In continuation of systematic research on flavonoids of plants in the Scutellaria L. genus (Lamiaceae) we have investigated the roots of Scutellaria schachristanica Zuzepezuk collected in the Samarkand region of Uzbekistan, in the flowering phase. By the column chromatography of an alcoholic extract of ethylacetate fraction on silica gel, we isolated a 0 flavonol derivative with the composition C16H12O5 (I), mp 215-217 C. Compound 1 was analyzed by IR, UV (with NaOAc) and 1H NMR spectroscopy. From the spectral date and by double resonance its structure was deduced as 5,7-dihydroxy-3- metoxyflavone. 5,7-dihydroxy-3-metoxyflavone first time has been isolated from plants of the genus Scutellaria. Alcoholic extract of S. schachristanica showed high antioxidant activity.

HO O

OCH3 OH O

65 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Optimization of Method of Isolation of Leucomisine from Artemisia leucodes Schrenk

Vladimir V. Folmer, Ilya A. Khabarov, Bibigul B. Rakhimova, Svetlana A. Ivasenko, Anar N. Zhabaeva, Khorlan I. Itzhanova, Sergazy M. Adekenov

JSC “International research and production holding “Phytochemistry”, 100009, Karaganda, Kazakhstan Fax: (7212) 43-37-73 E-mail: [email protected]

The original hypolipidemic preparation “Aterolide” is developed on the basis of sesquiterpene lactone leucomisine of guaiane type isolated from Artemisia leucodes Shrenk. at the holding „Phytochemistry“. At the present, the base of technology of the leucomisine’ isolation from Artemisia leucodes is an extraction of raw material and chromatographic purification with use of classical method – column chromatography, which is needed of use of great quantity of toxic organic solvents. Therefore the aim of our investigation is development of effective and ecologically safe method of isolation of leucomisine from raw material of Artemisia leucodes. We carried out approbation of method of supercritical fluid extraction of plant raw material of Artemisia leucodes for the obtaining of substation of preparation “Aterolide”. Technological process of CO2–extraction of herb of Artemisia leucodes is optimized, namely influence of temperature, pressure, duration of extraction on the quantitative content of purposing substance in CO2–extraction. Dependence of degree of isolation of leucomisine from Artemisia leucodes is studied by changing of pressure from 100 to 300 atm, temperature from 40 to 600C and duration of extraction from 120 to 240 minutes. Investigations on selection of conditions for isolation of leucomisine from CO2–extract of Artemisia leucodes is carried out with the use of preparative high-performance liquid chromatography, in particular selection of stationary and mobile phases. It is tested three types of immobile phases which can provided reversed-phase separation: carbon adsorbents, C18 modified silicagels, super-linked polymeric adsorbents. It is experimentally determined that use of column, fulled by Chromalite 5HGN, 5 m, as stationary phase and mixtures acetonitrile – water (80:20) as mobile phase is optimum for preparative isolation of leucomisine from CO2–extract of Artemisia leucodes. As components of CO2–extract of Artemisia leucodes is good separated one another from other by use of these conditions of chromatography, this is allowed to isolate purposing substance with purity no less 99,0 %. Besides mixture acetonitrile – water in ratio 80:20 cannot be separated at distillation, solvent returned in cycle with minimal losses. Thus the effective and ecologically safe method of isolation of leucomisine from raw material of Artemisia leucodes with use of supercritical fluid extraction and preparative high- performance liquid chromatography is developed. This method is characterized by comparatively high productivity, full automation, great shortening of duration of the process.

66 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation

Preparation of -Pinene Oxide from Turpentine

Sergey S. Laev, Vladislav V. Fomenko, Nariman F. Salakhutdinov

N.N. Vorozhtsov Institute of Organic Chemistry, 9 Ac. Lavrentjev Avenue, Novosibirsk, 630090, Russia E-mail: [email protected]

Terpenes are widely distributed in nature, and their epoxides often serve as starting materials for the synthesis of fragances, flavors and therapeutically active substances. Hydrogen peroxide is an efficient oxidant and epoxidation agent for terpenes if it is appropriately activated. Among the literature procedures of preparation of valuable -pinene oxide it should be noted the oxidation of -pinene by 30% hydrogen peroxide (10 eq) in the presence of catalyst, MnSO4, in DMF [1].

O H2O2

Now we report the simple, economical and technological preparation of -pinene oxide using our previous achievements connected with various terpene epoxidation [2]. The epoxidation of turpentine (content of -pinene is 75.6%) by 35% hydrogen peroxide (10 eq) in aqueous acetonitrile catalyzing by the system of MnSO4, NaHCO3 and salicylic acid led to a mixture containing -pinene oxide, residuary -pinene and minor components. Pure - pinene oxide was obtained via vacuum distillation of this mixture (yield ~30%). This route of -pinene oxide preparation from turpentine is more effective than expensive and diffucult separation of -pinene from turpentine followed by epoxidation..

1 Lane B.S., Burgess k. J. Am. Chem. Soc. 2001, 123, 2933-2934 2 Bakhvalov O.V., Fomenko V.V., Salakhutdinov N.F. Pat. RU №2005140845/04, №2006138540/04(041991).

67 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Chemical Research of Marsh Cinquefoil (Comarum palustre) Grows in the Krasnoyarsk Territory

Sergey V. Golubev, Alexander A. Efremov

Siberian Federal University, Ave. Svobodny 79, 660041, Krasnoyarsk, Russia E-mail: [email protected]

Marsh Cinquefoil (Comarum palustre) is perennial plant with creeping stem, grows in damp, swampy areas of Siberia. It is used in folk medicine and in the production of drugs. In this research provided data on the content of some classes of extractives, ash elements, composition of polysaccharide complex and essential oil. Collection of marsh cinquefoil was performed in July-August 2009 in the territory Emelyanovsky region of the Krasnoyarsk territory. Analysis of ash showed uneven distribution of inorganic substances in the plant. Using XRF (APL ADVANT'X intelliPower 3600) characterized the main inorganic components of Marsh Cinquefoil. Using method of exhaustive extraction with quantitative chemical analysis by electron spectroscopy in UV and visible spectral region showed that all parts of the plant contain different amounts of extractives, chief among which are: tannins, chlorophyll A and chlorophyll B, carotenoids, and vitamin C. It investigated polysaccharide complex. In addition, all parts of the plant contain essential oil (not more 0,1 %). Using GC-MS (Agilent 7890 A) showed that the main component is benzyl alcohol.

Table. Components of Marsh Cinquefoil Part of plant Components rhizome creeping stem green stem leaves collective fruit Ash content, % 1,70 2,24 5,07 6,95 7,18 ASS*, % 9,3 8,3 5,2 14,0 4,1 ESS*, % 14,8 16,3 7,1 15,4 10,4 WSS*, % 16,7 16,1 21,5 27,2 21,6 Chlorophyll A, mg% 3,9 7,8 21,4 179,2 24,9 Chlorophyll B, mg% 2,5 3,3 5,6 64,4 6,5 Carotenoids, mg% 2,5 3,5 6,5 71,4 7,5 Tannins, % 12,1 12,8 14,7 16,5 17,3 Vitamin C, mg% 130 142 152 149 192 WSP*, % 1,6 0,9 1,4 3,1 2,5 Pectins, % 1,9 1,5 1,9 1,6 12,6 Hemicelluloses A, % 2,3 3,0 14,9 16,7 1,9 Hemicelluloses B, % 25,2 18,9 19,3 23,0 21,2 Celluloses, % 32,6 35,8 29,5 18,9 23,2 Lignins, % 21,1 22,2 15,0 16,7 19,4 Inorganic components, K (1,89), Ca (0,89), Mg (0,42), P (0,29), F (0,05), Na (0,04), Mn (0,04), Si % (0,02), Fe (0,02), Zn (0,01), Al (0,01)

* − Acetone-soluble, ethanol-soluble, water-soluble substances, water-soluble polysaccharides.

68 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Ultrasound Assisted Enzymatic Hydrolysis of Cellulose as a Method of Improvement of Biologically Active Substances Extraction

O.V. Golyazimova1,2, A.A. Politov1,2, O.I. Lomovsky2

1Novosibirsk State University, Research and Educated center “Molecular Design and Ecological Safe Technology”, Pirogova, 2, Novosibirsk, Russia 2Institute of Solid State Chemistry and Mechanochemistry, Kutateladze, 18, Novosibirsk, 630128, Russia, e-mail: [email protected]

One of the ways of herbal raw materials application is the extraction of different organic physiological active substances, alkaloids, enzymes, hormones, et al, and production of medicines and food supplements. The conventional method of biologically active substances obtaining is its extraction with organic solvents. It is known that biologically active substances are situated in plant cells with thick and firm cell wall. The main component of plant cell wall is cellulose. To promote extraction process it is possible to hydrolyze cellulose with enzymes. It was found that preliminary ultrasound treatment accelerates this enzymatic reaction. Previously we have shown the acceleration of enzyme reaction by simultaneous ultrasonic treatment. Our studies are consistent with those obtained by other researchers. Then we revealed that preliminary ultrasonic treatment of enzyme – substrate system at temperature 0 ?C enhance the following hydrolytic reaction at 50 ?C, fig 1. It is interesting ultrasonic treatment of cellulose in water without enzymes is ineffectively, fig 1 (graph 2). The possible mechanism of reaction acceleration by ultrasound is its influence on enzyme sorption processes through the acoustocapillary effect.

25

20

, % % , - 1 15 - 2 - 3

10 Conversion 5

0 0 1 2 3 4 5 6 t, hrs

Fig. 1. Hydrolysis kinetics of cellulose: 1 – untreated cellulose, 2 – after ultrasound treatment without enzymes, 3 – after ultrasound treatment with enzymes.

This work was supported by CRDF (RUX0-008-NO-06) and ISTC № 3235.

69 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Mannich-Type Three Component Condensation of α-Substituted Caran-4-one Oxime with Formaldehyde and Secondary Amine

Nikolay B. Gorshkov, Alexandr M. Agafoncev

Novosibirsk Institute of Organic Chemistry, Lavrentjev Ave., 9, 630090, Novosibirsk, Russia Fax (838) 330-97-52 E-mail: [email protected]

The Mannich reaction of carbonyl compounds is a well-known synthetic method for the preparation of a variety of nitrogen-containing molecules. The aza derivatives of carbonyl compounds are also known as substrates for the Mannich-type condensation to yield β-amino- substituted derivatives. Chiral α-substituted oximes derived from natural terpenes are of great importance for coordination chemistry as polyheteroatomic ligands having an open-chain or cyclic topology of the set of donor atoms, and Mannich-type condensation could be a prospective approach to the design of new polydentante ligands. Here we report on the fist successful stereoselective introduction of an additional donor moiety into chiral α-substituted oximes by a three-component Mannich-type reaction

NHMe2-CH2O morpholene-CH2O MeOH-AcOH-H2O MeOH-AcOH-H2O S 80 oC, 24 h S 80 oC, 24 h S

N N N OH OH OH N N

O

NHMe2-CH2O MeOH-AcOH-H2O S S 80 oC, 24 h S S

N N N N OH OH OH OH N N

NHMe -CH O 2 2 NHMe2-CH2O MeOH-AcOH-H O 2 MeOH-AcOH-H2O o NR R 80 C, 24 h 1 2 80 oC, 24 h tar-like product N N R1 = H, Alk R1 = H OH R2 = Alk OH R2 = H N

70 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Quantitative Determination of Betulin and Lupeol in Birch Bark Extracts by HPLC

Alexander E. Grazhdannikov, Ekaterina A. Khokhrina, Olga P. Sheremet

N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry of the Siberian Branch of Russian Academy of Sciences, 630090, Novosibirsk, Lavrentiev av. 9 e-mail: [email protected]

Triterpene compounds of lupane structure containing in a birch bark are of interest for researches of biological activity and also synthesis of new medical products. Betulin (betulinol), lupeol and their compounds are attractive objects for the purposes of synthesis. GC analysis of birch bark extracts has been so complicated because of a very high boiling point of triterpene compounds. So HPLC method has been applied to the analysis allowing to determine the compounds of various nature without a preliminary analytic samples modification. The factors of sensitivity have been calculated for a quantitative determination of lupeol at a stage of betulin isolation. The factor klupeol/betulin to be calculated on a wavelength (λ) 210 nm is equal 0.8. To control of betulin and lupeol content in extracts two variants of gradient HPLC technique have been developed on the devices of a series «Milichrom» with the Nucleosil C- 18 reversed phase columns: 1) The variant «methanol-water (19:1, v/v) – methanol» have been used to analyse the betulin and lupeol contents in the technical birch bark extracts. 2) The variant «methanol-water (4:1, v/v) – methanol» has been used to analyse the betulin at a purification stage. Methylursolate has been applied as an internal standard for betulin and lupeol quantitative determination. This technique can be used in the scientific purposes and for monitoring technological processes including material balance investigations to develop the chemical process designing.

71 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Unusual Reactions of (+)-2- and (+)-3-Carenes with Aldehydes on K10 clay

Irina V. Il’ina, Konstantin P. Volcho*, Dina V. Korchagina, Georgi E. Salnikov, Alexander M. Genaev, Elena V. Karpova, Nariman F. Salakhutdinov

N.N.Vorozhtsov Novosibirsk Institute of Organic Chemistry, Siberian Branch, Russian Academy of Sciences,Lavrentiev ave., 9, 630090 Novosibirsk, Russia Fax: +7 383 3309752 E-mail: [email protected].

(+)-2- and (+)-3-Carenes (1 and 2, respectively) are important optically active monoterpenes, wich are very promising substrates for asymmetric synthesis due to the presence of a bicyclic framework, a reactive double bond, and a dimethylcyclopropane fragment, creating considerable steric hindrances on one side of the molecule, these. We have studied for the first time the reactions of (+)-2- and (+)-3-carenes with aldehydes in the presence of montmorillonite K10 clay. It was shown that the major products of these reactions were substituted chiral benzofuranes, probably formed as a result of an attack of protonated aldehyde at the cyclopropane ring. Quite unexpectedly, these products had cis-condensation of rings.

It appeared that the behavior of the 2 : 3 mixture of (+)-2- and (+)-3-carenes in reactions with aldehydes in the presence of clay K10 differed substantially from the reactivities of the corresponding individual monoterpenes. For example, using a mixture of (+)-2- and (+)-3- carenes in reactions with aromatic aldehydes led to considerably increased yields of the products formed from (+)-2-carene.

The authors are grateful to the Presidium of Russian Academy of Sciences for financial support (complex program N 15.2).

72 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Chemical Transformation of Cycloartane Triterpenoids

I.M. Isaev, D.A. Iskenderov, M.A. Agzamova and M.I. Isaev

S.Yu.Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax (99871) 120 64 75, e-mail: [email protected]

The syntheses on the basis of cyclosiversigenin (1) and cycloorbicoside A (7) have been conducted. The natural triterpenoids cycloasalgenin (2), cycloadsurgenin (4) and their 17- epimers 3 and 5, and lactone of cyclosiversigenin (6) have also been synthesized. The chemical transformation of cycloorbicoside A (7) has been realized in three directs. The structures of the synthesized compounds have been confirmed by methods of 1H, 13C NMR spectroscopy, DEPT, IR spectroscopy and mass spectrometry. OH OH

H H

O O

OH O H

HO HO OH OH 1 2. 17-H OH 3. 17-H

H O

O O

O OH H

O HO OH OH 4. 17-H 6 5. 17-H H H H H

O O OH O O OH

O OH O O O O OH 7 OH 8 HO HO OH OH H H H H

O O OH O O

O OAc O OH O O OH 9 OH 10 HO HO OH OH

73 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Perspectives of Ecdysten Use in the Treatment of Giardiasis as Mono- and Co-infection

Jannat I. Islamova, Vladimir N. Syrov, Zainab A. Khushbaktova, Svetlana O. Osipova

Institute of Chemistry of Plant Substances of Academy of Sciences of the Republic of Uzbekistan, Tashkent 100170, str. M. Ulugbek 77, Uzbekistan E-mail: [email protected]

Negative influence of giardiasis on macroorganism is stipulated not only etiopathogenetic characteristics (diarrhea, functional disorders of hepato-biliary system, asthenia, allergic manifestations, deceleration of mental and physical development of children) but also its ability to exacerbate the course of chronic bacterial (tuberculosis) and viral (HIV-infection) infections as a concomitant disease due to opposite type of immune protective reactions and worsening hepatotoxicity of the preparations applying for the treatment of the main disease. Treatment of giardiasis is complicated by increasing resistance of the parasite to conventional antigiardial preparations, it especially frequently occurs in immunodeficient organism. Ecdysten is a preparation developed on the basis of phytoecdysteroid ecdysteron, isolated from Rhaponticum carthamoides. Use of ecdysten in the treatment of giardiasis as adaptogen (5 mg 4 times daily for 10 days) showed its high efficiency exceeding 90% in primary and as well as in resistant to metronidazole, albendazole and other antigiardial preparations clinical form. Eliminations of parasites in >50% of patients was observed for the first five days of the course and was of dose-depending manner: in sporadic cases of the absence of elimination an increase of daily dose up to 25 mg resulted in eradication of parasites. Ecdysten was established to normalize decreased indices of cellular immunity and remove allergic manifestations. Ecdysten efficiently eliminates Giardia lamblia in patients with pulmonary tuberculosis and HIV-infected individuals, improving a general state of patients and tolerance to antituberculosis preparations and HAART, in particular it was demonstrated in normalization of activity of AlAT, AsAT and alkaline phosphatase and stabilization / improvement of immunologic indices. Thus ecdysten can be considered as an efficient preparation in the treatment of giardiasis including treatment against the background of chronic infections requiring prolonged therapy.

74 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Investigation of Capillary-Protective Composition on the Base of Dihydroquercetin and Lipoic Acid

Ivan S. Ivanov, Anna M. Anischenko, AnastasiaV. Sidekhmenova, Mark B. Plotnikov

Institute of Pharmacology, Siberian Department, Russian Academy of Medical Sciences, Lenin street 3, 634028, Tomsk, Russia Fax: 8(3822)41-42-35 E-mail: [email protected]

The aim this research is the stimation e of capillary-protective activity of dihydroquercetin, lipoic acid and their composition. Experiments were carried out on 30 male Wistar rats (250-300 g). The rats received dihydroquercetin (20 mg/kg), ipoic l acid (20 mg/kg) and compositions of dihydroquercetin/lipoic acid (correspondingly in doses of 20/10 mg/kg, 20/20 mg/kg and 20/40 mg/kg) p.o. daily for 7 days. Capillary-protective effect was studied using the method of I.A. Oivin. The criterion of capillary permeability was the exit time of Evans blue in inflammatory tissue. This colorant was administered intravenous in dose of 2 mg/kg (1% solution) 10 min before the administration of xylol (0,02 ml, intradermally). Course p.o. administration of dihydroquercetin increased the exit time of coloring material in inflammatory tissue by 27% (p<0,05) as compared to value in the control rats. Lipoic acid didn’t enhance significantly the exit time of Evans blue. Compositions of dihydroquercetin/lipoic acid in doses of 20/10 mg/kg, 20/20 mg/kg and 20/40 mg/kg increased the exit time of coloring material in inflammatory tissue by 30%, 51% and 13% (p<0,05) correspondingly in comparison with the control values. Additionally the effect of dihydroquercetin and lipoic acid in doses of 20 and 20 mg/kg was by 19% (p<0.05) higher than effect of dihydroquercetin. Thus, composition of dihydroquercetin (20 mg/kg) and lipoic acid (20 mg/kg) manifests distinct capillary-protective activity. Effect of composition exceeds the ffect e of dihydroquercetin.

75 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Investigation of Biological Activity of the Stereoisomeric Halogenhydrines of Ludartine

Roza I. Jalmahkanbetova

JSC “International research and production holding “Phytochemistry”, 100009, Karaganda, Kazakhstan Fax: (7212) 43-37-73 E-mail: [email protected]

The wide spectrum of biological activity of sesquiterpene lactones attract more attention of researchers in plane of search of the new compounds with more expressed pharmacological activity. The number of the sesquiterpene lactones with different types of activity every year

increase [12-3], such as antimicrobial, cytotoxic, anti-inflammatory and other types of activities. We carried out the test of the stereoisomeric halogenhydrines of sesquiterpene lactone ludartine 1 in ratio of grampositive and gramnegative strains. The antibacterial activity of the compounds was studied on 4 species of bacteria (Staphylococcus aureus 505, Bacillus subtilis, Escherichia coli M-17, Pseudomonas aeruginosa) and one type species of fungus (Candida albicans).

Cl HO Br HO O H H H H H O HO O Cl O HO O Br O 1 O 2 O 3 O 4 O 5 O

In the result it was determined the studied stereoisomer halogenhydrines 2-5 show the moderate expressed antibacterial activity. It was found the antibacterial activity of the stereoisomers considerably do not differ. In comparison with the initial lactone 1 the antibacterial activity of the synthesis compounds 2-5 by ratio to strains Bacillus subtilis, Escherichia coli M-17, Candida albicans are increased to 1,2-1,5 times.

1 Djeddi, S.; Karioti, A.; Sokovic, M.; Stojkovic, D.; Seridi, R.; Skaltsa, H. J. Nat. Prod. 2007, 1796-1799. 2 Bruno, M.; Rosselli, S.; Maggio, A.; Raccuglia, R.; Bastow, K.; Lee, K. J. Nat. Prod. 2005, 68, 1042-1046. 3 Feltenstein, M.; Schqhlyb, W.; Warnicka, J.; Fischerb, N.; Sufka, K. Pharmacology, Biochemistry and Behavior. 2004, 299-302.

76 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis of Tetrahydroquinoline-Barbituric Acid-Terpene Hybrids

Aliaksei P. Kadutskii and Nikolas G. Kozlov

Laboratory of Organic Catalysis, Institute of Physical Organic Chemistry, The National Academy of Sciences of Belarus. Surganov st., 13, 220072, Minsk, Belarus. Fax: (375 17)2841679 E-mail: [email protected]

A promising area of modern organic chemistry, awaiting further development, is the creation of hybrid molecules (hybrid molecules are defined as chemical entities with two or more structural domains having different biological functions and dual activity).1 Tetrahydroquinoline family compounds are used as a parent compound to make drugs, fungicides, biocides, catalyst ligands and dyes. A large number of barbituric acid derivatives have been widely used as sedative, anticonvulsant and local anaesthetic drugs as well as in antitumor, anticancer and antiosteoporosis treatments. It is known the pharmacological and physicochemical properties of certain drug classes can be modified by incorporating compact lipophilic moieties such as the fenchyl, adamantyl and bornyl groups. In this context, we report herein our methods for the synthesis of tetrahydroquinoline- barbituric acid-terpene hybrid molecules.

H O O N O R H H EtOH R terpene HN NH NH ketones NH O O O N O

R1 R1 1 2 3 4 R = CH3 , OCH3 , Ph

O R1 = ; ; ; ; ;

The synthesis includes two steps and starts from readily available terpene ketones, such as camphor, verbanone, etc. They were first converted in the corresponding arylamines 1 by the standard reductive amination protocol. These amines, on treatment with formaldehyde 2 and barbituric acid 3 in refluxing EtOH, undergo spirocyclisation to give the tetrahydroquinoline-barbituric acid-terpene hybrids 4.

1. Meunier, B. Acc. Chem. Res. 2008, 41, 69-77.

77 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation One-pot Synthesis of 3-Aminoacridine Derivatives

Aliaksei P. Kadutskii and Nikolas G. Kozlov

Laboratory of Organic Catalysis, Institute of Physical Organic Chemistry, The National Academy of Sciences of Belarus. Surganov st., 13, 220072, Minsk, Belarus. Fax: (375 17)2841679 E-mail: [email protected]

Acridine and its derivatives are basic substances for the production of dyes. They are also used as a starting material for the production of some drugs. Acridine is used for vital and supravital staining for the detection of nucleic acids, differentiation of DNA and RNA, and detection of malignant cells. Aminoacridine derivatives display interesting chemical and biological properties in the ield f of antitumor agents. 3-Amino- and 3-alkylamino-4- hydroxymethylacridines bearing various substituents on the C ring show potent cytotoxic activities against murine L1210 (leukemia), human A549 (lung), and HT29 (colon) cancer cell lines.1 In this contribution we propose a new method for one-pot synthesis of 3-aminoacridine derivatives. The polysubstituted acridines 1 were prepared in good yield by the one-pot reaction of N1,N1-dimethylbenzene-1,3-diamine 2, aromatic aldehydes 3 and cyclic - diketones 4 in alcoholic solvents (Scheme).

R1 R1 H R1 N NH2 N N R EtOH R1 O O O Reflux, 5 - 10 min R O 2 3 4 1 75 - 85 % R = Ar ; R1 = H , CH3 Scheme

This three-component reaction proceeds under mild conditions in the absence of an additional acid catalyst. A possible mechanism and some regiochemical, synthetic and applied aspects for the reaction will be presented.

1. Charmantray, F.; Demeunynck, M.; Carrez, D.; Croisy, A.; Lansiaux, A.; Bailly, C.; Colson P. J. Med. Chem. 2003, 46, 967–977.

78 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Obtain and Properties of Cryogel of Polyvinyl Alcohol

A.A. Kasymbekov, M.T. Omyrzakov, B.M. Kudaibergenova, Sh.N. Zhumagalieva, M.K. Beisebekov, Z.A. Abilov

The Al-Farabi Kazakh National University, Chemical department Almaty, Karasai batyr str., 95a E-mail: [email protected]

Cryogels on the basis of polyvinylalcohol (PVA) represent the big interest both in the scientific plan, and in applied. Besides, composite of macroporous polymeric cryogels are used as sorbents for sewage treatment from heavy metals. Also they have found wide application in biotechnology, medicine, the food-processing industry, etc., as a biomass, artificial fabrics, and carrier of medical drugs. In many respects this interest is caused excellent mechanical, diffusion and thermophysical by properties cryogels of PVA and availability of the polymer, nontoxic biocompatibility and gel formation ability. In this connection, possibility of reception cryogels on the basis of PVA by the method of cryogel formation and regularity of immobilization of local anaesthetic of richlokain on them has been investigated. 10, 13 and 15 % cryogels of PVA have been obtained and their swelling ability was studied. By the results of investigation with increase in concentration of PVA, the degree of swelling () of cryogels raises, also makes for 4,5-7,9. For an estimation of possibility of use cryogels as carriers of medicinal substances was carried out immobilization of local anaesthetic of medical drug – richlokain on data cryogel by the method of sorption. By the results of studying the limiting values of sorption of richlokain on gels within 2 day make up 40-55 % that testifies about enough high sorption abilities. In all studying substantial growth of degree of sorption richlokain with increase of concentration PVA that it is possible to explain according to the rule of acting weights has observing. Therefore, the size of sorption makes from 3,05 mg/g – 4,2 mg/g. In that way, as a result of investigation was obtained cryogels of PVA. And their sorption ability under the attitude of medical drug – richlocain was considered.

79 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation

Isolation of Arglabin from CO2-Extraction of Artemisia glabella Kar. et Kir. by Use of Flash-Chromatography

Ilya A. Khabarov, Svetlana A. Ivasenko, Bibigul B. Rakhimova, Sergazy M. Adekenov

“International research and production holding “Phytochemistry”, 100009, Karaganda, Kazakhstan Fax: (7212) 43-37-73 E-mail: [email protected]

Sesquiterpene lactone arglabin of guaiane type isolated from aerial part of Artemisia glabella Kar. et Kir. is an active base of new high-performance antitumor preparation “Arglabin” which developed at the holding “Phytochemistry”. As main stage of technology of the obtaining of arglabin is column chromatography, which is characterized by low speed of separation and needed of use of great quantity of toxic organic solvents. Therefore the aim of our investigation is development of effective and ecologically safe method of isolation of arglabin from CO2-extract of Artemisia glabella. We studied a possibility of use of flash-chromatograph «Millipore K-Prime 40-III» for preparative isolation of arglabin from CO2-extract of Artemisia glabella. Maximal submission of eluent is 500 ml in minute, maximal pressure is 7 atm, column of size 1100x100 mm. This variant is optimized with use of column, which filled up by adsorbent Chromalite 5HGN with size of particles 15 m, mobile phase: mixture of acetonitrile – water (80:20). At volume speed of submission of eluent 237 ml in minute the complete separation of tested sample is resulted in the course of 2,3 hours, working pressure is 5 atm, temperature of column is room. In total 10 injections can made for day in fully automated mode, i.e. with quantitative isolation of arglabin with purity no less 99,0 % the separation of CO2-extract of Artemisia glabella is possible. Thus, approbation is carried out, and it is experimentally determined that the proposed method of preparative isolation of arglabin from CO2-extract of Artemisia glabella by use of flash-chromatograph «Millipore K-Prime 40-III» is characterized by comparatively high productivity, full automation, great shortening of duration of the process. The preparative isolation of arglabin with purity of no less 99,0 % by use of this method in fully automated mode for day.

80 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Ferula foetida (Bunge) Regel Growing in Uzbekistan

E.Kh. Khalilova, O.Sh.Yusupov, B.T. Sagdullaev, Kh.S. Akhmedkhodjaeva, N.Sh. Davlatov, S.F. Aripova

Akad. S.Yu. Yunusov Institute of the Chemistry of Plant Substances Academy of Sciences, 100170 Tashkent, Uzbekistan, fax (99871) 120 64 75, e-mail: [email protected]

Data about the distibution of the plant Ferula foetida (Bunge) Regel belonging to the family – Apiaceae (Umbelliferae) were given in the article. It has been noted that the dried juice of the roots of Ferula foetida is used in oriental forlk medicine as anti-inflammatory, analgetic, spasmolitic, and antiparasitary drugs for treatment of differend deseases. Toxicity and pharmacological activity (estrogenic) of the alcoholic extract from the plant Ferula foetida, collected in Buchara district, were studied on laboratory animals

81 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthetic Modification on the B Ring of Labdane-Type Diterpenoids

Yurii V. Kharitonov, Elvira E. Shults, Genrikh A. Tolstikov

N. N. Vorozhtzov Institute of Organic Chemistry, Siberian Branch of the Russian Academy of Sciences, Novosibirsk tel.: (383) 3308533, fax: (383)3309752 E-mail: [email protected]

Highly functionalized B ring labdane diterpenes are a very interesting class of natural products due to the important biological activity that some of them show1. We have been interested in the synthesis of Δ8-7-ketolabdanods which could be transformed into several preparations for medicine. As the starting compounds we have used lambertianic acid (1) and its methyl ether (2). Treatment of (1) or (2) with p-toluenesulphonic acid in benzene leads to compounds (3, 4) (yield 86% and 89% respectively). We have shown that the reaction of labdanoids (3) and (4) with SeO2 provides a selective introduction of a hydroxyl group on position 7β. Hydroxy derivatives (5) and (6) were obtained in the yield 59% and 74% respectively. The ketones (7) and (8) are obtained by oxidation of the indicate allylic hydroxy compounds with MnO2 (yield 49 and 69%). Synthesis of octahydro-1H-benzoazepine derivatives has been carried out by Beckmann rearrangement of oximes (9) and (10) (obtained from compounds 7 and 8 in the yield 89 fnd 98%). Treatment of labdanoid oxime (9) with thionyl chloride in dry dioxane led to the formation of substituted octahydro-1H- benzo[d]azepine (11) as the main product. Beckmann rearrangement of oxime (10) gave a mixture of substituted octahydro-1H-benzo[d]azepine9-carboxylic acid (12) and octahydro- 1H-benzo[c]azepine- 9-carboxylic acid (13).

O O O O SeO PTSA 2 MnO2

OH O H H H H RO2C RO2C RO2C RO2C 3, 4 1, 2 5, 6 7, 8

O O O NH2OH.HCl/ AcONa SOCl2 NH + O N OH H NH RO C H 2 H O 9, 10 HO2C HO2C 11, 12 13 (30%) R=H (1,3,5,7,9,11); CH3 (2,4,6,8,10,12)

This work was supported by the Russian Foundation for Basic Research (project no. 09-03-00183).

1 Hanson, J.R. Nat. Prod. Rep., 2009, 26, 1156.

82 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Studying Alkaloids of Haplophyllum griffithianum

D.R. Kodirova, H.A. Rasulova, R.Sh. Shakirov

Acad. S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, 100170,Tashkent, Republic of Uzbekistan, fax: (99871) 1206475 e- mail: [email protected]

The plant of genus Haplophyllum is known as medical for a long time. Many species of genus Haplophyllum and other genera family Rutaceae belongs to a number of less investigated, and some of them have not been studied on the alkaloid contents. Considering the above-mentioned, study of alkaloids of Haplophyllum represents certain theoretical and practical value. We have studied new species Haplophyllum griffithianum from two places of growth. Total alkaloids from the overground parts and roots of plant collected at Sanchardaksay (Nilu village) consisted of 0.4% and 0.58%. The total alkaloids from the overground parts and roots of Haplophyllum griffithianum, collected at Holdize (Surkhandarya region), were yielded 0.45% and 0.6% respectively. From the over ground part have been isolated for the first time the quinoline alkaloids – skimmianine, dictamnine, dubinine, and a new base with mp. 121-122° and from roots – dictamnine, skimmianine and sterens with mp. 76-78° and 60°. From the over ground part and roots of Haplophyllum griffithianum, collected in Holdize (Surkhandarya) have been allocated dubinine and skimmianine. From the overgroud part of Haplophyllum griffithianum for the first time we isolated a new dihidropiranoquinoline alkaloid with mp. 121-1220 and called it grifitinine. The structure of grifitinine was established on the basis of the spectral data and has the structure I. O

N O

CH3 I

83 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Chemical Fixation of Betulonic Acid on Water Soluble Polyvinyl Alc ohol

Tamara I. Kogay1,2, Tatiana L. Sostina2, Boris N. Kuznetsov1,2

1 Institute of Chemistry and Chemical Technology SB RAS, K. Marx str.,42, Krasnoyarsk, 660049 fax: +7(391)2439342, e-mail: [email protected]; [email protected] 2 Siberian Federal University, Krasnoyarsk

The low solubility of triterpene compounds in the water restricts their application in medicine. Different triterpene compounds are produced by chemical transformations of the birch bark betulin. Betulonic acid (BA) obtained by betulin oxidation has various physiological activity including anti-HIV activity. It is expected that the chemical fixation of betulonic acid on water soluble polyvinyl alcohol will result to the formation of biologically active BA-containing polymer dissolved in the water. The reaction of polyvinyl alcohol acylation by chloric anhydride of betulonic acid was used for BA fixation on polymer in the form of ester:

O H2C CH3 +Bet C C Cl (CH2 CH) (CH2 CH) n n Bet = CH3 -HCl CH3 OH O COOH C Bet O O CH3 CH H3C 3 The influence of reaction conditions on the degree of polyvinyl alcohol acylation was studied. It was found that the use of phase-transpher catalyst triethylbenzyl- ammoniumchloride and dichlormethane or benzene as organic phase makes it possible to produce the water- soluble form of betulonic acid – polyvinyl alcohol polymer with the yield 45-47 % at temperature 18-20°C and time 2 hours. IR spectra of acylation products, dissolved in water have absorption bands at 1804 cm–1, corresponding the valence vibration of C=O group in esters, at 1705 cm–1 corresponding to stretching vibrations of keto-group of betulonic acid and at 1454 cm–1, belonging to valence vibrations of C-H bonds of cyclohexane ring of betulonic acid. In order to prove the presence of ether bond between betulonic acid and polyvinyl alcohol the alkaline hydrolysis samples was studied. Adsorption bonds in IR-spectra of polymer at 1804 cm–1, 1705 cm–1 and 1454 cm–1, corresponding to ester bonds and to betulonic fragment were disappeared in spectra of hydrolysed samples of polymer. This experimental fact shows that the water-soluble samples of acylated polyvinyl alcohol contain the chemically bonded betulonic acid in the form of ester fragment.

84 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis of New Esters of Peoniflorin, the Major Monoterpene Glycoside of Paeonia anomala L. Roots

R.M. Kondratenko1, Z.G. Kulmukhametova2, O.V. Stolyarova2, L.A. Baltina (jr.)2, L.A. Baltina1,2

1Bashkir State Medicinal University, Lenin’s stree, 3, 450000, Ufa, Russia; 2Institute of Organic Chemistry Ufa Research Centre of RAS, prospect Oktyabrya, 71,450054, Ufa, Russia Fax: 7(347)2356066 E-mail: [email protected]

Bioactive metabolites of Paeonia (P.) roots especially peoniflorin (PF) to be the major monoterpene glycoside of P. anomala L. roots attract the high attention of chemists and pharmacologists in the last decade due to different kinds of biological and pharmacological activities. PF is known by hypotensive and miotropic activities, antioxidative and antitrombic properties and stimulates the mind’s abilities. The content of PF in Paeonia anomala L. roots (Russia) is about 2% and it is easily separated from extracts by chromatography. Chemistry of PF is slightly studied and its synthetic transformations and synthesis of new derivatives are the actually task. We firstly synthesized 4-O-alkyl esters of PF by the reaction of 4-OH group with low molecular alcohols. Reaction of PF with MeOH or EtOH proceeds at the reflux in the excess of corresponding alcohol in the presence of cation exchange resins (H+). The yield of 4-O- methylglycoside was 84.8%. Synthesis of 4-O-Bu ester of PF was carried out in the presence of HCl. The NMR 1H spectrum of 4-O-Me ester of PF contains the singlet of OCH3 at 4.0 ppm, and 13C NMR – 51.4 ppm. NMR spectra of other 4-O-alkyl esters contain additional signals of CH2 groups. 4-O-alkyl esters were transformed into full acetates by the use of acetic anhydride-pyridine mixture.

4-S-alkyl esters of PF were synthesized by the reaction of PF acetate with CH3CH2SH or

CH3(CH2)5SH in the presence of BF3 (48-50% yields).

HO O CH 3 O HO O HO OH O COOCH OH

2

(1) This work was supported by the grant of SS 1725.2008.3.

85 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation

Synthesis of New Chiral Ligand from (+)-2-Carene

Ekaterina A. Koneva, Konstantin P. Volcho, Yuri V. Gatilov, Dina V. Korchagina, Nariman F. Salakhutdinov

N.N.Vorozhtsov Novosibirsk Institute of Organic chemistry SB RAS 630090, Lavrentiev Av., 9, Novosibirsk, Russia e-mail: [email protected]

The reaction of (+)-2-carene with chlorosulfonylisocyanate led to the tricyclic lactams corresponding to the initial formation both of the tertiary and β-cyclopropylcarbenium ions. The lactams were used for the synthesis of the optically active -amino acids derivatives and salicylaldimines – new chiral Shiff bases from (+)-2-carene.

86 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation The New Method of Salicylates Extraction

Anna V. Koptina, Alexey I. Shurgin, Albert V. Kanarskiy

Mari State Technical University, Lenin sq., 3 424000 Yoshkar-Ola, Russia Fax:0078362410872 E-mail: [email protected]

Willow (Salix spp.) is characterized by high amount of low molecular weight phenolic glycosides such as salicin and its derivatives, which have been shown to be biologically active substances. Salicin is an active precursor for salicylic acid and therefore willow bark, which contains a high amount of total salicin, is used in phytotherapy of many countries as an analgesic, anti-inflammatory and antipyretic drug1,2. Furthermore, the metabolites within the willow extract may act as tumor inhibitors that promote apoptosis, cause DNA damage, and affect cell membranes and/or denature proteins.3 The goal of this research was to develop a new method of salicylates extraction from willow bark that will allow to increase the yield of biologically active substances and to prevent degradation of labile compounds. According to the goal, the new extraction method was invented and optimized. Air dried willow plant material was grinded on the particles less than 2 mm in diameter, and extracted with potassium phosphate buffer (pH 5.5) at hydromodule 50 and temperature 65±5°C during 9 hours. The enzyme complex of cellulase and β-glucanase was added to destroy the plant cell walls consisting mostly of carbohydrates (cellulose, hemicelluloses etc.). Described mode increases the yield of salicylates up to 12 %. Furthermore, the bark after extraction can be used as phytosorbent due to the presence of many active centers.

1 ESCOP monographs on the medicinal uses of plant drugs; Fascicule 4, 1997. 2 Julkunen-Tiitto, R. J Nat Prod 1992, 55, 1204-1212. 3 El-Shemy, H. A.; Aboul-Enein, A. M.; Aboul-Enein, K. M.; Fujita, K. PLoS One 2007, 2, e178.

87 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Triterpenoids of Picea and Pinus Species

Tatyana P. Kukina, Emma N. Shmidt

Novosibirsk Institute of Organic Chemistry, Siberian Branch of the Russian Academy of Sciences, 9 prosp. acad. Lavrent’ev, 630090, Novosibirsk, Russian Federation E-mail: [email protected]

In the recent review, the literature data on the phytochemical and biological investigations on the triterpenoids and sterols of Abies genus are summarized. Up to now, more than 100 triterpenic compounds were isolated from plants of Abies species. The chemical constituents of this group are mostly lanostane triterpenoids. The crude extracts and triterpenic metabolites have been found to possess various bioactivities including antitumor, antimicrobial, antiulcerogenic, anti-inflammatory, antihypertensive and other. The presence of lanostanoids is the feature of Abies genus on Pinaceae family. Present review are summarized the data about the triterpenoids of other genuses of this family. Principal feature of Picea genus is the high contence of serratane triterpenoids in raw material. A lot of articles describes the evaluation of serratane-type constituents from Picea species bark1. Some papers show the identification of new triterpenoid class on Picea plants. It is so-called abeoserratenes2. We obtained a sum of serratenes from needles of Picea obovata. The identification of these constituents continues3. These results are very interesting in sum with data about cancer chemopreventive activity of serratane-type triterpenoids on two-stage mouse skin carcinogenesis4, multi-organ carcinogenesis5. Some serratene were studied for their possible inhibitory effects on Epstein–Barr virus early antigen activation induced by 12-O- tetradecanoylphorbol-13-acetate (TPA). The result of the present investigation indicated that these compounds might be valuable as a potent cancer chemopreventive agent. Some lanostane triterpenoids were obtained from Picea species, but content of these constituents is less than in Abies raw materials6. The Pinus raw materials include as lanostane as serratane too7. A number of triterpenoid compounds evaluated from bark and needles are more than 20 substances8. This data show the great potencial of Pinaceae family raw materials for progress of medicine.

1 Chernenko, G.F.; Shmidt, E.N.; Chemistry for sustainable development, 1997, 5, 95-103. 2 Tanaka, R.; Tsujimoto, K.; In, Y. et al. Tetrahedron, 2002, 58, 13, 2505-2512. 3 Shmidt, E.N.; Chernenko, G.F.; Kukina, T.P.; Pokrovskii, L.M. Chemistry of Natural Compounds, 2008, 44, 4, 508-609. 4 Tanaka, R.; Tsujimoto, K.; Minami, T. et al. Cancer Letters, 2001,172, 2, 119-126. 5 Yamaguchi, Ch.; Wanibuchi, H.; Kakehashi, A. Food and Chemical Toxicology, 2008, 46, 6, 1882-1888. 6 Kutney, J.P.; Rogers, I.H.; Rowe J.W. Tetrahedron, 1969, 25, 17, 3731-3751. 7 Fang, J.M.; Tsai, W.Y.; Cheng Y.S. J. Chin Chem Soc., 1991, 38,1, 61-64. 8 Kutney, J.P.; Eigendorf, G.; Worth, B.R. Helvetica Chimica Acta, 1981, 64, 4, 1183-1207.

88 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis of Betulin Esters by Interaction of Organic Acids with Birch Bark

Svetlana A. Kuznetsova1,2, Galina P. Skvortsova1, Boris N. Kuznetsov1,2*

1 Institute of Chemistry and Chemical Technology SB RAS, K. Marx str.,42, Krasnoyarsk, 660049 fax: +7(391)2439342, e-mail: [email protected]; [email protected] 2 Siberian Federal University, Krasnoyarsk

Known multistage methods of betulin esters synthesis are based on reactions of acylation of betulin isolated from birch outer-bark. New methods of betulin diacetate, dipropionate and dibutirate synthesis directly from birch bark are described in this presentation. It was found that the treatment of birch outer- bark by acetic, propionic and butyrate acids results to integration of stages of betulin extraction from bark and its acylation by organic acid according to the scheme:

R: -CH3 – for betulin diacetate, -CH2-CH3 – for betulin dipropionate, -CH2-CH2-CH3 – for betulin dibutyrate.

Synthesis of betulin esters was carried out at boiling temperatures of organic acids. After reaction the solution was concentrated on rotor evaporator and diluted by five-fold amount of water. The precipitated sediment was filtred, washed by distillated water and dried on the air. Obtained products were recrystallizated from ethanol in the presence of active carbon. In order to select the optimal conditions of synthesis, which ensure the maximal yield of betulin esters, the following parameters of bark treatment by organic acids were varied: liquid to solid ratio from 10 to 30 and time of acylation from 0.5 to 18 hours. The yield of betulin esters was between 30-42 % from mass of a.d. birch outer-bark. Melting temperatures of recrystallizated betulin diacetate, dipropionate and dibutyrate were 222 °C, 150-152 °C and 63-65 °C, respectively. For identification of betulin derivatives the methods of elemental analysis, FTIR and NMR H1 and C13 were used. Obtaned betulin esters have prospects for the use in medicine and in synthesis of other biologically active derivatives of betulin.

89 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation New Methods of Allobetulin Synthesis

Vladimir A. Levdansky1,2, Svetlana A. Kuznetsova1,2, Galina P. Skvortsova1, Alexander V. Levdansky1, Boris N. Kuznetsov1,2*

1 Institute of Chemistry and Chemical Technology SB RAS, K. Marx str.,42, Krasnoyarsk, 660049 fax: +7(391)2439342, e-mail: [email protected]; [email protected] 2 Siberian Federal University, Krasnoyarsk

Allobetulin (19,28-epoxyoleanan-3-ol) pentacyclic triterpene has various biological activity. The known multistage methods of allobetulin synthesis are based on the extraction of betulin from birch outer bark and following isomerization of betulin to allobetulin in the presence of acid catalysts. It was suggested to produce allobetulin by isomerization of ethanol extract of birch outer bark with H3PO4 catalyst. This makes possible to exclude the stage of betulin isolation and purification. The total conversion of betulin presented in birch bark extract to allobetulin is near 30 hours. The purity of allobetulin is increased after the addition of active carbon to betulin- containing extract. In this case the yield of allobetulin reaches to 39-40 % from mass of air- dry birch outer bark. The one-stage method of allobetulin synthesis from birch outer-bark was developed. This method is based on the integration of betulin extraction and its acidic isomerisation to allobetulin. The treatment of bark impregnated by H2SO4 or H3PO4 in boiling toluene or xylene gives the rather high yield of allobetulin. The influence of concentration of sulphuric acid catalyst and time of treatment on the yield of allobetulin was studied. The maximal yield of allobetulin (2 % from mass of abs.dry bark) was obtained with the use of 40-50 % H2SO4 for bark impregnation and time of treatment in boiling solvent near 3 hours. Table. Influence of conditions of birch bark treatment on the yield of betulin Sample Concentration of H SO for bark 2 4 Time of treatment Yield of allobetulin No impregnation, % Solvent toluene 1 30 4.5 20 2 40 4.0 21 3 50 3.5 22 4 60 3.0 19 5 70 3.0 18 Solvent xylene 6 30 4.0 21 7 40 3.0 22 8 50 2.5 21 9 60 2.0 18 10 70 2.0 17 Melting temperature of obtained allobetulin is 275-277 °C. For identification of allobetulin the methods of elemental analysis, FTIR and NMR H1 and C13 were used.

90 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Palladium-Catalyzed Amination in the Synthesis of Diamino Furocoumarin Derivatives

Alla V. Lipeeva, Elvira E. Shults, Genrikh A. Tolstikov

N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, SB RAS, Novosibirsk, Russia

Peucedanin (1) is a available natural furocoumarin. It is isolated from Peucedanum Morissonii in the yield about of 4% 1 [ ]. Compound (1) was quantity transformed into bromooreoselone (2) by the action of bromine in CCl4. Bromination of (2) with NBS in the presence of benzoyl peroxide led to the formation of 2-(1,3-dibromopropan-2-ylidene)-2H- furo[3,2-g]chromene-3,7-dione (3). This compound was subject o t palladium-catalyzed amination with various amine and amino acid derivatives. The results of these reactions were found to strongly depend on the catalyst and amine nature. Amination of (3) with L-(+)- valinol using Pd(dba)2–Ph3P as the catalyst system [2] gave the diamino furocoumarin (4) in low yield. Reaction of 3 () with L-(+)-valinol under the catalyses by Pd(OAc)2–o- tolylphosphine in DMF solution in the presence of triethylamine as a base gave the compound (4) in the yield of 45%. To use of BINAP as a ligand in the reaction of (3) with L-(+)-valinol increases the yield of diaminocoumarine (4) to 72%. Compounds (6-8) were obtained by the palladium catalyzed reaction of (3) with 2-aminoisobutyric acid, 2-(1-piperidino)aniline, L- phenylalanine methyl ester or ethylenediamine. Reaction of (3) with 1,6-diaminohexane gave the corresponding macrocyclic diamino compound (9).

NH O N O HO HN H2C O O O O NH 9(63%) HN HN H2C O O O CH2 6(44%) ii O N i,ii HO H2C O O ii 4 (45, 72%) O NH BrH2C HOOC H BrH2C O O O N O ii H3C OCH3 3 H C HN ii 2 O O ii HOOC H C O O 2 O O H N O H3C 5(35%) OH NH2C H O H3C O O O H2NH2CH2C HN H2C H O O NH C 2 H2NH2CH2C HN H2C O O O NH 7(65%) i: Pd(OAc)2/(o-tol)3P OH 8(38%) ii: Pd(OAc)2/BINAP O O

This work was supported by the Russian Foundation for Basic Research (project no. 08-03-00340).

1 Shults, E.E.; Petrova, T.N.; Shakirov, M.M., Chernyak, E.I.; Pokrovsky, L.M.; Nekhoroshev, S.A.; Tolstikov, G.A. Chem. Sust. Dev., 2003, 11, 695. 2 Averin, A.D.; Uglov, A.N.; Ranyuk, E.R.; Lukashev N.V., Beletskaya, I.P. Rus. J.Org. Chem., 2009, 45, 280.

91 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation The Flavonoids of Some Species of Kazakhstan Rumex L.

Y.A. Litvinenko, R.A. Muzichkina

Kazakh National University named by al-Farabi, Chemistry Department, Str. Karassay batir 95 a, Almaty, Kazakhstan E-mail: [email protected]

There are about 150 species of plant genus Rumex L. in the world’s flora, from which 49 species of the flora of the USSR and 23 species of Kazakhstan are described. Studied plants of this family have a broad spectrum of biological activity and they are used in the folk and scientific medicine in different countries. The polyphenolic compounds (tannins, flavonoids, anthraquinones, phenols and phenolic acid) and other biological active substances dominate in the composition of most of them. We have studied all kinds of Kazakhstan plant genus Rumex L. and 4 kinds of Rumex L.: R. tianschanicus Los., R. thyrsiflorus F., R. rossicus Murb. and R. Marschalllianus R. chb. were included into officinal medicine. The roots of the 7 Rumex L. of Kazakhstan: R. thyrsiflorus F., R. rossicus Murb., R. Marschalllianus R. chb., R. maritimus L., R. pamiricus L., R. aquaticus L. and R. pseudonatronatus B., gathered in the phase of rest in Almaty, Kokshetau and Aktobe regions are used for comparison of the composition of flavonoids. The 50% aqueous acetone was the optimum extragent from 9 tested extragents all over the qualitative set of BAS and quantitative content of extractive substances for all kinds of the Rumex L. By the method of adsorbtion chromatography on the polyamide and silicagel using as eluents of water, ethyl alcohol, chloroform, system of water-ethanol in various proportions 6 flavonoids, which on the grounds physical and chemical constants in comparison with authentic samples and literature data were identified as isorhamnetin, quercetin, myricetin, myricetin-3-O-α-L-rhamnopyranoside, kaempferol-3-O-α-L-rhamnopyranoside and isorhamnetin-3-O-α-L-rhamnopyranoside. The total content of flavonoids in the studied species Rumex L. ranges from 2 to 10%. All kinds of Rumex L. contain quercetin and myricetin and they are the hemotaksons of the Kazakhstan species. R. thyrsiflorus F., R. pamiricus L., R. rossicus Murb. distinguish by the presence of myricetin-3-O-α-L-rhamnopyranoside, and R. thyrsiflorus F. contains the isorhamnetin and two 3-O-α-L-rhamnopyranosides of the isorhamnetin and kaempferol. Preferable places of gathering of 5 species of Rumex L. are Almaty region, R. Marschalllianus R. chb. Kokshetau region and R. rossicus Murb. Aktobe region.

92 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Solid State Mechanochemical Reaction of Hypericin in Plant Material

I.O. Lomovsky, O.I. Lomovsky

Institute of solid state chemistry and mechanochemestry SB RAS, Russia, 630128, Novosibirsk, Kutateladze str. 18, [email protected], Research Education Center ‘Molecular design and environmental safe technologies’ Novosibirsk state university, Russia, 633090, Novosibirsk, Pirogova str. 2

Biologically active substance which exhibits antioxidants properties plays one of the main roles in protection of normal vital functions of organisms. Current technologies of production of extracts and preparation from plants are based on processes used solvents with differ polarity and critical liquids. Because of antioxidants are reducers, they have strongly activity with oxidizers contains in extract. Decreasing of yield of antioxidants, nonreproductivity of concentrations of active compounds, problems with storage are technical effect of high activity of antioxidants. The purpose of our research is effort of ability of producing of antioxidant preparation with using of solid mechanochemical treatment. We have chosen the one of the most active and perspective compound – hypericin from the St. John’s Wort (Hypericum perforatum L.) as an example. Hypericin and related compounds have some hydroxyl groups with acid properties. The reaction of neutralization has been chosen for solid state onversion c of hypericin to biologically available state. As an alkali we used the number of solid compounds with increasing alkalinity: NaHCO3, Na2CO3, NaOH. The luminescence spectra of sum of naphtodiantrones adsorbed of solid cellulose depends on the alkalinity of adsorber. It is able us control the acid-base reaction of hypericin in solid plant matrix. It is known that a lot of organic compounds by the intensive mechanical treatment decay. The conditions of treatment when hypericin was not decay was experimentally chosen. By the treatment plant material was fine grinded up to size of particles 20 – 150 mkm. The solid alkali particles have size less than 50 nm. The producing of this nanocomposite powder provides conversion of hypericin and relative compounds in water-soluble form directly by the production in solid state or increases velocity and yield of dissolution by adding water, for example by the putting in stomach. In case of water extraction we saw the increasing of yield of hypericin in comparison with other extraction methods in 10 times. The maximum concentration of hypericin in mechanochemically produced preparations is the same as those which contents in plant material by exhaustive extraction. The comparative stability of hypericin preparations in solid and liquid phase was studied. Thus, we showed availability of using of solid phase mechanochemical reactions for producing of preparations of unstable antioxidant compounds from plant material. The velocity of oxidizing of active component – hypericin from St. John’s Wort in the solid mechanochemical preparation in comparison with those in liquid preparation was decreased not less than 30 times. This work was supported by grants of CRDF (RUX0-008-NO-06), RFBR (08-09-12130, 09-03-92658), and RAS Program “Fundamental Sciences for Medicine”

93 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Phytochemical study of Artemisia pontica L.

Darya L. Makarovaa, Minisa A. Khaninaa, Dmitry V. Domrachevb, Alexey V. Tkachevb

a Novosibirsk State Medical University, Krasnyj prospect, 52, 630091, Novosibirsk, Russia b Novosibirsk Institute of Organic Chemistry, 630090, Novosibirsk, Russia E-mail: [email protected]

Artemisia pontica L. (Asteraceae) has been known for centuries as Roman Wormwood and Petite Wormwood from Southern Europe, and nowadays it is found in eastern North America, Europe, southern Siberia and Central Asia. A. pontica is a prospective source of biologically active extracts and individual compounds and can be used for treating different inflammations and wounds. Most of research papers published on chemical study of A. pontica have been carried out using the plants from different regions of Europe. However, the European A. pontica has been mentioned to differ significantly from the plant of Siberian populations in sets of certain secondary metabolites. To perform a pharmacognosy study of A. pontica we collected the plants both growing wild in Novosibirsk and Altai regions and cultivated at the experimental plots in Tomsk and Novosibirsk (different stages of plants development: beginning of growth, budding and flowering; different parts of the plant: flowers, leaves, stems). Air dried material was subjected (a) hydro-distillation to afford essential oil and (b) solvent extraction to get polyphenols and other polar compounds. Hydro-distillation of the aerial parts of A. pontica resulted in 0.6–1.7 % of essential oil with the maximum at the beginning of flowering (1.7 %), the highest yields were from flowers (2.3 %), the lowest – from stems (0.06%). GC-MS study of the essential oil shows up to 200 components, the main constutuents are: 1.8-cineol (up to 70 %), camphor (up to 26 %), borneol (up to 17 %), vulgaron B (up to 39 %), chamazulene (up to 19 %). All the samples studied were dark blue due to the presence of chamazulene. Chamazulene has not been reported as a component of essential oils from European populations of A. pontica, and that is the main feature pointing the difference in biosynthesis of sesquiterpene lactones which are known as natural azulenogenic compounds. Polar extracts from A. pontica contains phenolic acids (chlorogenic, hydroxyl cynnamic, and caffeic acids, etc.), flavonoids (rutin, quercetin, apigenin, luteolin, etc.), water-soluble polysaccharides (up to 9.3%), pectin-type substances (up to 7.1%), hemicelluloses A (up to 13.6%) and hemicelluloses B (up to 3.7%).

94 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Extractive Compounds of Punica granatum L. Flowers

N.M. Mamatkulova, F.A. Melikuziev, S.Z. Nishanbayev, N.K. Khidyrova, S.F. Aripova, Kh.M. Shakhidoyatov

Institute of the Chemistry of Plant Substances, Academy of Sciences, Republic of Uzbekistan, Mirzo Ulugbek str., 77, Tashkent. Fax: 998-71-120-64-75 E-mail: [email protected]

Pomegranate (Punica granatum L.) – subtropical fruity bush, resistant to air dryness, widely spread in the Central Asia, Caucasus, Dagestan and Pamir–Altai. By now pomegranate fruits have been researched, where ascorbic and pholic acids, R– active catechines and leucoantocyans, vitamins B1, B2, tannin and pectin compounds, and also microelements were found. It is known that, leafs, fruits and barks of pomegranate is rich with tannin compounds. In the current study for researching the chemical content of extractive compounds of Punica granatum L. flowers, hexane, ethylacetate and n–butanol and water extracts. it were obtained by serial extraction. Alkaline hydrolysis of hexane extract was carried out and obtained non–alkaline compounds with the yield of 20 % out of total. Primary analysis showed that, the flowers of pomegranate contain polyisoprenoid alcohol, formerly found in flowers of Malvaceae family.

95 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Dissolution Rate Control of Nitrogen Containing Forage Additives

Serguey G. Mamylov1, Oleg I. Lomovsky1, Victor A. Rogachev2, Vladimir A. Soloshenko2

1 Institute of Solid State Chemistry and Mechanochemistry SB RAS, ul. Kutateladze 18, Novosibirsk, 630128, Russia 2 Siberian Scientific and Technological Institute of Husbandry SB RAAS, Krasnoobsk Novosibirsk region, 630500, Russia E-mail: [email protected]

Actual cattle-breeding problem in Siberia is to supply protein fodder. From this point of view, carbamide (urea) represents considerable interest. Urea is used usually in mixture with concentrated forages. But wide utilization of carbamide is limited by its high solubility and accelerated hydrolysis in ruminant digestive system. In this case, formed ammonia doesn’t utilize by proventriculus microflora. Considerable quantities of ammonia fall into bloodstream, resulting in chronic and sharp poisoning, in mortality of cattle. It is necessary to diminish urea rate of dissolution and hydrolysis for safety and effective using of carbamide in cattle feeding. Mechanochemical treatment allows control not only composition, but also properties of forage additions. The aim of present work is to investigate and to adjust dissolution rate of forage nitrogen- containing additives prepared by mechanochemical treatment of raw components. Mechanochemical treatment was carried out in planetary mills under controlled conditions. Various “grain-carbamide” mixtures were made containing bentonite (clay), straw, sunflower cake also. Urea dissolution rate from additives was investigated with the help of precision pH-metric installation. The dissolution of prepared forage additions is carried out in safety pH values range under conditions relative to the condition in cattle stomach. It became possible to vary initial rate of acidity changing ([H]/sec) up to hundreds times in comparison with the simple mixture. The optimal conditions of mechanochemical production of fodder additives were defined leading to the products with accessible rates of dissolution. Addition utilization in cows feeding occurs without gradual adaptation, the intoxication doesn’t take place, milk production is enhanced.

96 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Steroid Glycosides in Some Allium L. Species of Alliaceous Family (Alliaceae J. Agardh)

N.V. Matistov, T.I. Shirshova, I.V. Beshley

Institute of Biology of Komi Scientific Centre of Ural Division of Russian Academy of Sciences. Kommunisticheskya, 28, 167982 GSP-2, Syktyvkar, Russia Fax: (8212)24-01-63 E-mail: [email protected]

Steroid glycosides in Allium’s kin plants have been found not long ago. Nowadays there are more than 40 furostanol and spirostanol glycosides extracted from this plant kin, with its carbohydrate chains including D-glucose, a D-xylose, D-galactose, L-rhamnose and L-pectine sugar. From 26 genins discovered at the end of the last century the most common is diosgenin, found in 18 kinds of alliaceous, on the basis of which the medicinal preparations of a steroid constitution are produced. The most perspective among the studied species is the wild nodding onion A. nutans L. and the broadleaf wild leek A fuscoviolaceum Fomin, characterized by the high diosgenin content (2.1 and 2.3 % respectively). The analysis of the steroid glycosides content of nine alliaceous species from the Botanic garden collection of the Institute of Biology of Komi Science Center (RAS) – A. nutans L., A. narcissiflorum Wells., A. giganteum Rgl., A. jajlae Vved., A. komarovianum Vved., A. porrum L., A. angustifolium L., A. ramosum (odorum) L., A. schoenoprasum L. has proved the presence of steroid glycosides in all these onions [1]. For the first time the sum of steroid glycosides (SSG) from chive A. schoenoprasum has been isolated by 70 %-s' methanol extraction, their allocation in different parts of a plant has been shown. After SSG separation into close-cut fractions and individual materials by means of column and thin-layer chromatography, reversed-phase chromatography (RPC) and the received products analysis using specific reagents, it has been determined that received SSG contains from six to ten individual substances of spiristanol and furastanol nature. SSG has been divided into spiristanol (SG) and furastanol (FG) glycosides. From the sum of spirostanol glycosides using the column chromatography on alumina and silica gel two individual materials have been separated. Analysis by means f o a thin-layer chromatography, reversed-phase chromatography (RPC), and chromatography mass spectrometry has allowed identifying one of them as deltonin – the spirostanol glycoside firstly received from roots of deltoid dioscorea Dioscorea deltoidea [2]. It is found that diosgenin is genin SG which has been found both in an unbound state, and in products of acid hydrolysis of sum SG. From sum FG after gel chromatography on Sephadex G-25, separations into a chromatographic column, furostanol glycoside have been secured two – deltoside and protodioscine, identified by means of a thin-layer chromatography, RPC and hromato-mass spectrometry.

1 Shirshova T.; Volkova G.. Rast. resorts 2006, 42, 3, 59 – 66. 2 Pasechnichenko V., Vasil’eva I. Priklad. biochem. and microbiolog. 1995, 31, 1, 73-79.

97 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Sterols as a Part of Neutral Lipids in Some Allium L. (Alliaceae) Species

N.V. Matistov, T.I. Shirshova, I.V. Beshley

Institute of Biology of Komi Scientific Centre of Ural Division of Russian Academy of Sciences. Kommunisticheskya, 28, 167982 GSP-2, Syktyvkar, Russia Fax: (8212)24-01-63 E-mail: [email protected]

Complex research of chemical composition of three kinds of onion – A. angulosum L., A. schoenoprasum L. and A. strictum Schrad has showed, that their green material has a valuable connatural combination of vitamins, essential highest fatty acids, the vital micronutrients and others biologically active materials [1, 2]. Neutral lipids (NL) are one of the important ingredients of all parts of a plant, secured with triple extraction by hexane. The maximum content of NL is revealed (to 9 %) in seeds and decreases in a range: cover scales (~ 5 %) – knops (~ 3 %) – roots (~ 1.85 %) – leaves (~ 1.3 %) – bulbuses (~ 0.7 %). By a thin-layer chromatography method it has been found that essential ingredients of all parts of a plant, with the exception of seeds, are sterols and their Aethers, free fatty acids and their Aethers, threacyleglyceride (TAG). NL of seeds generally contain TAG. Using the gas-liquid chromatography of methyl Aethers of the highest fatty acids received by methanolysis NL, it has been discovered that they have acids with chain length from C16 to C20. Polynonsaturated highest fatty acids (PHFA), linoleic (C18:2) and linolenic (C18:3) are cores with total amount obtaining 60 – 75 %, depending on a part of a plant, relative to total PHFA. By means of column chromatography NL separation on silica gel (eluent – hexane with an increasing gradient of ethoxy ethane) sterine fraction (SF) has been isolated. The structure of SF has been found by a method of reversed-phase chromatography (RPC) after solid-phase extractions on the concentrating Diapak C18 cartridge and has been confirmed by a chromate- mass spectrometry method. Using RPC method three representatives of sterols have been found – Sitosterin, cholesterol and a stigmasterol. Chromate-mass Spectrometric research has allowed to find the fourth ingredient – kampisterol. The main component is sitosterin followed by kampisterol, a stigmasterol and cholesterol.

1 Shirshova, T.; Beshley, I; Gruzdev, I. Rast. resorts 2008, 44, 1, 75-81.

2 Shirshova, T.; Beshley, I. Rast. resorts 2009, 45,2, 97-105.

98 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Substance from Plant Limonium Myrianthum as the Basis for Creation of New Medical Products

Kseniya Mayeva, Anastassiya Gadetskaya, Zhanar A. Kozhamkulova, Galiya E. Zhusupova, Zharylkasin A. Abilov

al-Farabi Kazakh National University, Karasai Batyra 95a, 050012, Almaty, Kazakhstan E-mail: [email protected]

An evident tendency for wider use of medical products of plant origin in the official medicine can be observed in the leading countries of the world lately. The share of such kind of medical products from the total number of released remedies makes up more than 50 %. In this connection, providing the population with effective, ecologically clean and cheap medical products, obtained on the basis of approved by folk medicine plant material, is one of the main social policy priorities of the government of the Republic of Kazakhstan (RK), directed to the formation of our own pharmaceutical industry. Plants of the Limonium Mill genus, Plumbagenaceae family, presented in the world flora by 300 species, and in Kazakhstan by 18 species, may be placed among the most valuable herbs. Two species – L. gmelinii and L. myrianthum have industrial stocks on the territory of RK, which exceeds 54.4 thousand tons. On the basis of the roots of L. gmelinii tincture “Limonidine” (Temporal pharmacopeial article – TPA RK 42-1820-07) was developed, possessing high antiinflammatory, healing, antimicrobial, antiviral and hepatoprotective activities (Pharmacopeial article – PA RK 42-1259-08). On its basis ointment “Sanzhar” (PA RK 42-1277-08) and syrup “Limonidine” (TPA 42-1819-07) were developed. For rational and complete use of natural resources such studies were carried out for L. myrianthum as well. Moisture of the upper part of L. myrianthum equals to 7.5 %, of the roots to 11.2 %; total ash (upper part – 6.4 %, roots – 4.7 %), sulfated ash (upper part – 6.1 %, roots 6.1 %), ash, unsoluble in 10 % HCl (upper part – 0.7 %, roots – 0.4 %). All esteemed parameters indicate qualitativeness of raw material and low content of mineral substances in it. By varying solvent’s nature, temperature and time mode of extraction, as well as its multiplicity, optimal technological schemes for isolation of substances in form of dry extracts from the upper parts and the roots of the studied plants were elaborated. Qualitative values of the obtained substances were used as the principal criterias while working out optimal conditions for their isolation. As a result 50 % ethanol was chosen as the most optimal solvent. Plant material: solvent ratio equaled 1:8 for the upper part and 1:6 for the roots, extraction lasted for 24 hours under temperature of 22-25 0C with double repetition of the whole process of extraction. High antioxidant activity of the obtained substances was determined, which might be connected with the high content of tannins (11.27 % in the roots, 6.14 % in the upper part), flavonoids (5.84 % in the roots, 4.56 % in the upper part) and organic acids (0.36 % in the roots, 0.15 % in the upper part). Antioxidant activity of the obtained substances, isolated from both the roots and the upper parts are comparable to that of -tocopherol, well known natural antioxidant.

99 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Preparation and Solid-State Characterization of Solvates of Betuline and Its Mechanocomposites with Polymers

M.A. Mikhailenko1,2, T.P. Shakhtshneider1,2, V.A. Drebushchak2,3, T.N. Drebushchak1,2,, S.A. Kuznetsova4, V.V. Boldyrev1,2

1 Institute of Solid State Chemistry and Mechanochemistry SB RAS, Novosibirsk, Russia, e-mail: [email protected] 2 Research and Education Center "Molecular Design and Ecologically Safe Technologies", Novosibirsk State University, Novosibirsk, Russia 3 Institute of Geology and Mineralogy SB RAS, Novosibirsk, Russia 4 Institute of Chemistry and Chemical Technologies SB RAS, Krasnoyarsk, Russia

Biological activity of betuline and its derivatives as antiviral, anti-inflammatory, antitumour agents is under intense investigation at present. However, the physicochemical properties of betuline are insufficiently well studied. Any data on polymorphous and pseudo- polymorphous (solvate) forms of betuline are absent from literature. The authors [1, 2] of report different melting points for betuline samples extracted using different methods and recrystallized from different solvents, which may point to he t existence of several polymosphous modifications of betuline or its solvates with organic solvents2. Investigation of the polymorphism of medical and biologically active substances is known to be the necessary stage in the development of new dosage forms [3]. The goal of the present work was to investigate the possibility of obtaining different crystalline forms of betuline, as well as obtaining and investigation of the properties of betuline compositions with polymeric supports. The results of study of the products of crystallization of betuline from the solvents differing from each other in polarity (acetone, methanol, ethanol, propanol, butanol, ethyl acetate, chloroform, dichloromethane) are presented. The solvates of betuline were obtained. The methods of thermal analysis, X-ray diffraction and IR spectroscopy were used to investigate their composition and the process of their desolvation. It was shown that crystallization from the solvents used and the subsequent drying at a temperature of 170- 180oC allowed us to lower the lupeol content in the samples down to less than 1%. One of the methods to increase the biological availability is to obtain compositions with water-soluble polymers, in particular, with the help of mechanical activation. The stability of betuline under mechanical activation was demonstrated. By means of the joint mechanical activation of betuline with polymeric supports, stable compositions were obtained; their dissolution gives higher concentrations than betuline dissolution does. The formation of complexes and solid solutions of betuline with polyvinylpyrrholidone was demonstrated by means of IR spectroscopy and thermal analysis.

1 Komissarova N.G., Belenkova N.G., Spirikhin L.V., Shitikova O.V. and Yunusov M.S. // Chemistry of Natural Compounds. 2002. V. 38. P. 58-61. 2 Hayek E.W.H., Jordis U., Moche W. and Sauter F. A // Phytochemistry. 1989. V. 28. P. 2229-2242. 3 Byrn S.R., Pfeiffer R.R. and Stowell J.G. Solid-State Chemistry of Drugs. SSCI, West Lafayette, IN, 1999.

100 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Reaction of (-)-Cytisine with Thiourea

A.V. Minilbaeva, A.N. Lobov, I.P. Tsypysheva*, M.S. Yunusov

Institute of Organic Chemistry of Ufa Research Center of Russian Academy of Science Russia, 450054, Ufa, prosp. Octyabrya, 71. E-mail: [email protected]

Reactions of secondary amines with urea and thiourea are completely studied, but it is surprising that well-known quinolizidine alkaloid (–)-cytisine was not involved in this direct interaction – only a few carbothioamides were obtained by alternative arylisothiocyanate way1-3. As an extension of “thiourea methodology” to the alkaloids chemistry area, we present a first example of (–)-cytisine carbothioamides synthesis on the basis of thiourea and (–)- cytisine 1 to result desirable carbothioamide 2 and unusual product 3 – thioanalogue of bimolecular alkaloid argentine (scheme 1). Compound 2 was isolated by crystallization to yield 48%. S S NH N NH2 N N N thiourea + N N N

O 1 O 2 O 3 O Scheme 1

The product 2 structure (fig.1) was elucidated at laboratory for X-ray Diffraction Stadies of A.N. Nesmeyanov Institute of Organoelement Compounds Russian Academy of Science. Fig. 1

Compound 3 was isolated by column chromatography on silica gel to yield 16%. The structure of 3 is according to NMR spectroscopic and elemental analyses data.

This study was financially supported by the Russian Foundation for Basic Research (project № 09-03-00685-a).

______

1. G. Luputiu, Archiv der Pharmazie, 1969, 302, 943-945. 2. O. Nicolotti, C. Canu, F. Sparatore, A. Carotti, Farmaco, 2002, 57, 469-478, 3. C. Canu, B. Tasso, V. Boido, F. Sparatore, Farmaco, 2003, 58, 265-278

101 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Naphthoquinone Substituted Labdanes via Diels-Alder Reaction

Maksim E. Mironov1,2, Elvira E. Shults1, Yurii V. Kharitonov1, Genrikh A. Tolstikov1

1N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, SB RAS, Novosibirsk, Russia 2Novosibirsk State University,Russia, e-mail: [email protected]

Labdane type diterpenes are of interest as the starting compounds for the synthesis of fragrances and pharmaceuticals1. We focused our attention on lambertianic acid (1), which can be readily isolated from cedar oleoresin and needles. Several transformations of this metabolite with the formation of biologically active compounds were carried out2. Herein we describe the synthesis of lambertianic acid derivatives, containing a naphthoquinone substituent. Reaction of 16-formyl labdatrienoate (2), which was obtained from (1) by the successively of methylation, isomerization and Vilsmeier-Haack reactions, with acetone in the presence of potassium hidroxyde in water results in the condensation product (3) (yield 67%). The reaction of compound 3 () with chlorotrimethylsilane affords the labdanoid silyloxybutadiene (4) in high yield. The behavior of the newly synthesized siloxybutadiene in the Diels-Alder reaction has been studied using quinones as dienophiles. The reaction with p- benzoquinone in seeding benzene leads to the formation of a labdanoid with a naphthoquinone moiety (65%). The cycloaddition of (4) with substituted quinones proceeds with high yield but low regioselectivity. So, in the reaction of diene (4) with toluenequinone compounds (6) and (7) are formed in the ratio of 10:7. The influence of the catalyst on the ratio and yields of the product has been studied.

1 Chinou, I. Curr. Med. Chem., 2005, 12, 1295. 2 Shults, E.E.; Velder, J.; Schmalz, H.-G.; Chernov, S.V.; Rubalova, T.V.; Gatilov, Y.V.; Henze, G.; Tolstikov, G.A.; Prokop, A. Bioorg. Med. Chem. Lett., 2006, 16, 4228; Kharitonov, Y.V.; Shults, E.E.; Sorokina, I.V; Tolstikova, T.G.; Baev D.S.; Zukova, N.A.; Tolstikov, G.A. Rus. Patent. No 2353620. (Chem. Abstr. 2009, 150, P495008k).

102 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Alkaloids of Korolkowia Sewertzowi – Original Antiarrhythmic Drugs of Different Mechanisms of Action

Yu.R. Mirzaev, R.Sh. Shakirov, D.U. Abdullaeva

Akad. S.Yu. Yunusov Institute of the Chemistry of Plant Substances, 100170 Tashkent, Uzbekistan

From aerial and underground parts of Korolkowia severtzowii plant (Liliaceae fam.), endemic for Central Asia have isolated more than 50 individual bases belonging to different subclasses of steroidal alkaloids. Main alkaloids – korseveriline, sevedine and sevedinine form around 50% of total sum of plant alkaloids. Latter related to C-nor,D-homosteroidal group of steroidal alkaloids. Indeces of their acute toxicity on white mice at i.v. administration expressed by numbers 235, 180 and 257 mg/kg. Compounds antaginised to aconitine called arrhythmia on narcotised white rats with ED50 12,6; 9,4 and 5,7 mg/kg i.v. correspondly. Indeces of pharmacological width of antiarrhythmic activity (K=LD50/ED50) of mentioned compounds were expressed by the numbers 18,7; 19,1 and 45,1. These numbers for known antiarrhythmic drugs quinidine, procainamide and rhythmilene were 5,8; 5,7 and 16,6 correspondly. Besides, sevedinine revealed antagonism to fibrillation called by CaCl2 on narcotisd rats. It is known, that mentioned classic antiarrhythmics of 1-st class, especially rhythmilene, have nonspecific M-cholinoblocking activity on heart, smooth muscle and secretory organs, including CNS effects. This fact provide number of different side effects on mentioned organs. In opposite to classic antiarrhythmic compounds, Korolkowia alkaloids block only M-receptors of heart, that belongs to M2-subtype, without other M1,3,4 type receptors touching. This is important difference of Korolkowia alkaloids compared with mentioned classic antiarrhythmic drugs. Sevedinine on isolated ventricle of guinea pig prevents positive inotropic and positive chronotropic effects of isoproterenole and it is may be evaluated as β1-adrenoblocking activity of compound. LD50 of semisynthetic 3- acethylcevedine on white mice was 125(91?174) mg/kg. ED50 on aconitine model – 4,6±1,8 mg/kg, index K – 17,6. LD50 of semisynthetic 3,6-diacethylsevedine and 3,6- diacethylkorseveriline were 41(32?52) and 81(72?92) mg/kg, E D50 on aconitine model 2,2 and 4,5 mg/kg i.v., indeces K – 17,6 and 18,6 correspondly. Diesters of sevedine and korseveriline were low active on CaCl2 model of ibrillation f and they loss M2- cholinoblocking activity. All Korolkowia alkaloids in low degree supressed electrostimulated papillary musle of guinea-pig contractility and arterial pressure compared with known mentioned classic drugs. Thus, Korolkowia alkaloids belongs to different classes of antiarrhythmic drugs. Diasethylsevedine and diacethylkorseveriline may be declared as antiarrhythmics of 1 class, korseveriline and sevedine belong to I+III classes and sevedinine is composed antiarrhythmic of I+III+IV classes. In clinical practice composed antiarrhythmics may be useful. Sevedinine is pecial s remarkable compound suitable for regulating pharmacological neurogenic denervation of heart.

103 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation The Study of Qualitative and Quantitative Characteristics of the Kazakhstan Type of Plant, Polygonum Alpinum

A.E. Mussina, R.A. Muzichkina

KNU named by al-Farabi, chemistry department, str.Karasaibatyr 95”a”, Almaty, Kazakhstan Fax:8(727)2590946 E-mail: [email protected]

From the number of the 123 species of highlanders described in the flora of the USSR, there are 49 grow in Kazakhstan, but only 4 of them are pharmacopoeian and approved by TPA RK. Polygonum has a number of useful properties: antituberculosis, diuretic, antiulcer, anti-inflammatory and bleeding, anticancer, tonic. The area of distribution of Wild Highlanders is very vast, in particular polymorphic species Polygonum Alpinum, also known as Polygonum Undulatum Murr or Polygonum Polymorphum. The chemical composition of this type will determine the possibility of its use in the pharmaceutical industry of the Republic. In laboratory terms, refer to The State Pharmacopoeia XI, have been identified the moisture content in raw – 1.44%, total ash – 6.50%. The elemental composition of Polygonum Alpinum, some atomic-absorption method, is presented in the table (%):

1 2 3 4 5 6 7 8 9 10 11 Pb Fe Ni Mn Cu Ca Mg K Cd Zn Na

6,84*10–4 1,04*10– 7,32*10–4 2,35*10–2 1,03*10– 1,54 7,31*10– 3,12 4,02*10–5 9,38*10– 2,95*10–

From the table macro-K, Ca, Mg elements are dominated. Qualitative and quantitative analysis of Polygonum Alpinum identified the dominant biologically active substances: a mixed-type tannins, flavonoids (quercetin, miritcetin, avikulyarin, katehin), phenolic acids (gallic, coffeic, chlorogenic, sirengic), phenols (pyrogallol, floroglucine, resorcine), aminoacids (Dl-triptofan, prolyne, α-glutamyne), organic acids, polisaccarids, saponins, alkaloids. The sum of polyphenols – 9.45%, polysaccharide – 3,045%, carboxylic acid – 0,31%, alkaloids – 0,074%, tannins method of permongonatometry – 6,84% (2,85% hydrolyzed and 3,99% of the condensed types) and method of complexonometry – 6,03%. These results indicate that the Kazakh Polygonum species may have high biological activity and be in the one line with other examples of pharmacopoeia. Fine-tune manufacturing mode extraction of biologically active compounds for fitocomplex: selected optimal solvents-50% alcohol and 50% dioxane with a ratio of raw material to solvent 1:6 and 1:8, respectively. The extractives limit for 50% alcohol – 24.51%, for 50% dioxane – 16,45%. The number of effective extractions uninterrupted mode with temperature of boiling of water bath is 2.

104 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Correction of Hypoxia of States at Animals Individual and Integral Materials from the Plant Convolvulus subhirsutus

A.N. Nabiev, S.F. Aripova, A.M. Gapparov, V.U. Khujaev, M.U. Alikulova

Akad. S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, 100170 Tashkent, Uzbekistan, fax (99871) 120 64 75, e-mail: [email protected]

The hypoxia of a various genesis accompanies with series of morbid conditions. Search of agents with anti-hypoxia action is actual in modern medicine. The purpose of work. To study anti-hypoxia activity of integral materials and 12 individual alkaloids from plant Convolvulus subhirsutus on model histotoxic and hemit hypoxias, and also at immune depressant. Stuff and methods. Tested materials are received as the sum of alkaloids from an above- ground part, the sum of alkaloids from roots C. subhirsutus, convolvine, bis convolvine, convolicine, N-acetylchloride of convolvine, N-benzyl convolvine, N-hexyl convolvine, methyliodide of convolvine, of N-benzoyl convolvine, N-nonilat convolvine, N-iodo amylat of convolvine, N-nitrobenzoyl of convolvine. Pharmacological researches were carried out on 10 in each bunch. A hypoxia invoked an injection of diazotizing salt or Sodium nitroprussidum. Activie substance estimated on lifetime of animals after intoxication of hypoxant. Results and their discussion. Intoxication by diazotizing salt characterizes oxygen- carrying function of system of a blood supply. From tested bonds by the most awake were bis convolvine which enlarged lifetime of mice on 60,8 %, convolicine on 54,8 %, N- acetylchloride of convolvine on 50,5 %, N-benzyl convolvine on 38,6 %, convolvine on 31,7 %. The conclusion. So, anti-hypoxic active compounds were found among alkaloids isolated from the plant C. subhirsutus, and their derivatives.

105 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Obtaining and Hepatoprotective Action of the Drug from Codonopsis clematideae

A.N. Nabiev, B.T. Sagdullaev, S.F. Aripova, Z.A. Hushbaktova, V.N. Syrov

Akad. S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, 100170 Tashkent, Uzbekistan, fax (99871) 120 64 75, e-mail: [email protected]

Hepatoprotective drugs are used in various clinics (therapy, surgery, dermatology, etc.). The purpose of work was to investigate hepatoprotective action and to reveal the optimal technological algoritm of obtaining of drug from the plant Codonopsis clematideae. Object and methods. From the plant Codonopsis clematideae alcohol-aqueous extracts obtaining various methods. It was investigated hepatoprotective activity of a dry powder («Dang-shen») influence on cholagogue, processes of synthesis of bile acids, abjections of a bilirubin and a cholesterin in bile of rats. At an acute fatty dystrophies of a liver at rats caused

CCl4 determined activity ALT, nuclear heating plant, glycogen of a liver, lactescent and pyroracemic acids and also triglycerides and phosphotides. Results and discussion. «Dang-shen» at introduction to rats stimulated cholagogue on 36,9-23,3%. Synthesis of bile acids was activated and has made 1510 mg% against 1025 mg%. The secretion of a cholesterine has decreased up to 26 mg % against 32,5 mg % in the control. At an acute fatty dystrophia treatment by a drug «Dang-shen» promoted drop of superactivity ALT and AST plant on 24 and 31 %, respectively. For 7 and 14 days of treatment action of a drug on activity of cytolytic enzymes strengthened. In the same terms the maintenance{contents} of protein and a urea in a blood was enriched. Positive dynamics{changes} is marked and from the side of triglycerides and phosphotides. Drugs «Dang-shen» enriched detoxify function of a liver, shortened duration of hexonal dream at intoxication by perchloromethane or heliotrine. Thus the drug «Dang-shen» renders positive action on flow of the experimental hepatites caused by perchloromethane and heliotrine. It normalizes enzymatic activity of serum of a blood, parameters of protein, carbohydrate and lipide exchanges in a liver, detoxic function of liver. Under influence of a drug «Dang-shen» the antihypoxic function both on histic, and on an oxygenium – transporting part of energy supply activated.

106 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Influence of Steroidal Alkaloids on the Steroidogenesis at Rats

A.N. Nabiev, R.Sh. Shakirov, D.U. Abdullaeva

Akad. S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, 100170 Tashkent, Uzbekistan, fax (99871) 120 64 75

Steroidal alkaloids have wide pharmacological activity. The purpose of work. To investigate action of alkaloids imperialone, imperialine, edpetiline on a biosynthesis of bile acids and a cholesterin; N-oxide of imperialine, iodo methylate of imperialin: solasodin and solanidine on flow androgenous dependence changes in mice at a stimulation by horionic gonadotropinum. Material and methods. Dynamics of changes of bile acids and cholesterin was investigated in bile at rats. A stimulation of androgenic activity was replicated at mice by injection by horionic gonadotropinum. The contents of a cholesterin in blood, mass of a liver and intrinsic generative organs of mice were determined. It is investigated anti-hypoxic activity of steroidal alkaloids for revealing participation in the mechanism of influence on a steroidogenesis. Results and their discussion. Alkaloid imperialine enhanced the biosynthesis of bile acids and cholesterin, imperialon enlarged biosynthetic processes of formation only bile acids and did not variate process of formation of cholesterin. Edpetiline did not influence formation of bile acids and has activated process of formation of cholesterin. Influence on an exchange of male sexual hormones at mice has shown, that among N-oxide of imperialine, Iodo methylate of imperialine, solasodin and solanidine, solasodine was the most awake, which enlarged seed fluid in 2,5 times on a background of a stimulation by horionic gonadotropinum, decreased mass of a prostate In the mechanism of change of a steroid exchange the activation of an oxygenation of tissues and systems participates. Hence, steroidal alkaloids stimulate biosynthetic processes of formation of steroids at animals and have anti-hypoxic activity.

107 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis of Bromo- and Iodo-Berberine

Ivan V. Nechepurenko, Nina I. Komarova, Marina P. Polovinka, Nariman F. Salakhutdinov, Genrikh A. Tolstikov

N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry of the Siberian Branch of Russian Academy of Sciences Lavrentjev Ave. 9, Novosibirsk, 630090, Russia Fax: +7 (383) 3309752 E-mail: [email protected]

Arylhalogenides are well-known substrates for reactions with C-, N-, O-, S- nucleofiles, and also they could be bioactive materials. We have developed methods of direct introducing bromine or iodine atoms into the isoquinoline alkaloid berberine 1. For the first time 12-bromo-berberine 2 was synthesized from compound 1 by means of bromination by Br2 in acetic acid in 76% yield. Formerly compound 2 was obtained when dihydroberberin was oxidized by bromine [1]. It was found that iodination of 1 by such reagents as ICl, I2 + NaIO3 + H2SO4, KI + CAN, KI + H2O2 + H2SO4, KI + AcOOH, NIS +

TFA, HgO + AcOH + KI + I2 don't proceeds. However, the reaction of 1 with the HgO + TFA

+ KI + I2 system results to a mixture of iodination products, the major of which, 4-iodo- berberine 3, was isolated in 36% yield. The possibility of selective introduction of haloid atom to A or D cycle of berberine molecule enlarges significantly the synthetic potential of this compound.

O O

O O Br

12

Br2 D N AcOH,  N CH3O CH3O

OCH3 Cl 1 OCH3 Br 2

HgO + TFA

O O

O O

A

4 I HgOCOCF3 KI, I2

N N CH3O CH3O

OCH3 OCH3 I 3

1 Iwasa, K.; Lee, D.-U.; Kang, S.-I.; Wiegrebe, W. J. Nat. Prod. 1998, 61, 1150-1153.

108 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis and Antioxidant Activity of Alkylthiomethyl Derivates of Hydroquinone, Resorcinol and Pyrocatechol

Sergey L. Nesterovich, Elena V. Korobicina, Aleksandr E. Prosenko

Research Institute of Antioxidant Chemistry, Novosibirsk State Pedagogical University, 28 ul. Vilyuiskaya, 630126 Novosibirsk, Russian Federation Fax: +7 (383) 244 1856 E-mail: [email protected]

Recent studies conducted at the Research Institute of Antioxidant Chemistry NSPU, showed high antioxidant activity of hydroxybenzyl sulfides with low sterically hindered OH- group. The ease of synthesis such compounds and the availability of the starting reagents demonstrates the great promise of the selected search direction of highly polyfunctional antioxidants and the need for its further development. It is known that diphenols and their ethers have higher rates of interaction with peroxide radicals than phenols. On the other hand, diphenols as an active component included in the structure of molecules of many natural antioxidants. Introduction of alkylthiomethyl groups in the molecules of hydroquinone, pyrocatechol and resorcinol, may lead to the formation of compounds significantly superior to increase inhibition activity of its sulfur-containing analogues based on phenol. Furthermore compounds of this type almost not described in the literature, which makes them even more interesting object of study. In this regard, we have obtained a number of benzyl sulfides based on 1,2-, 1,3- and 1,4- dihydroxybenzene, as well as a study of their inhibition activity on the models of auto- oxidation of fats. R = H, Me OH 1 R = CH2SC12H25 1 RO (R )n R2 = H, Me, i-Pr, Cy 2 n = 1,2,3,4 (R )m m = 1,2

It was shown that the most of synthesized compounds significantly surpass the antioxidant activity of phenolic sulfides with similar structure and all benchmark antioxidants.

109 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation A Convenient Synthesis of 3- and 3,10-Functionalized Indolizino[8,7-b]indole Derivatives from Harmine

Zhangeldy S. Nurmaganbetov1, Elvira E. Shults2, Sergey V. Chernov2, Aibek Zh. Turmukhambetov1, Alexander V. Kazantsev1, Genrikh A. Tolstikov2, Sergazy M. Adekenov1

1 JSC “IRPH “Phytochemistry”, Karaganda, str. M. Gazaliev, 4, Kazakhstan; E-mail: [email protected] 2 Novosibirsk Institute of Organic Chemistry, SB RAS, Novosibirsk, RF; E-mail: [email protected]

Natural metabolites, including a system of indolizino[8,7-b]indole (alkaloids harmicidine, trichotomine), show the valuable pharmacological activity. This heterocyclic system has been used for the preparation of more complex alkaloids. Our attention was attracted with an opportunity of construction of the named heterocyclic skeleton from the accessible plant alkaloid harmine (1). Interaction of harmine (1) with phenacyl bromides (2a-c) led to formation of harminium salts (3a-c). By treatment with sodium hydroxide compounds (3a-c) were exposed to cyclization with formation of 11H-indolizino[8,7-b]indole derivatives (4a-c). O Br + N KOH 1 H3CO N N R H H3CO N R2 2a-c CH3 O H CH 3 EtOH 1

1 R2 R 7 6 8 5 9 3a-c 10 11 4 N H3CO N 3 H 1 2 1 2 1 1' 1 R = R = H (a), R = H, 3' R 2 1 2 4' R = OMe (b), R = R = Cl (c) 4a-c R2

POCl3/ DMF N CHO AcCl/ H3CO N 4b H Py CHO 4a N CHO H3CO N H CHO Ph 6 5 OCH3 We investigated an opportunity of introduction of functional groups in specified tetracyclic skeleton of indolizinoindole. So, under acetylation of compound (4a) with AcCl in pyridine a 3-acetyl derivatives (5) was obtained. Formylation of polycyclic compound (4b) by Vilsmeier led to formation of 3,10-bis-formyl-11H-indolizino[8,7-b]indole (6).

This work was financial supported of the integration project of SB RAS with academies of sciences of the CIS countries (project № 79).

110 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis of 3-Hydroxy-4-[hydroxy(phenyl)methyl]-3-methyl-3,4- dihydrofuro[3,2-c]coumarin

Irina A. Os’kina, Yurij V. Gatilov, Alexey Ya. Tikhonov

Novosibirsk Institute of Organic Chemistry, Acad. Lavrentiev Ave., 9, Novosibirsk 630090, Russia Fax:(383)3309752, E-mail: [email protected]

Furocoumarins take important place among natural compounds because of wide spectra of biological activities. These compounds can be produced from natural raw materials1 or 2-4 synthesised.23 Literature data concerning of synthesis of dihydrofurocoumarins have presented only a few references.3,4 We have established that -hydroxy 3 -4-[hydroxy(phenyl)methyl]-3-methyl-3,4- dihydrofuro[3,2-c]coumarin 1 was formed under interaction of 4-hydroxycoumarin with 2- acetyl-3-phenyloxyran. This reaction is a new route for preparation of 3,4-dihydrofuro[3,2- c]coumarins.

O O

OH O O O

Et3N + O Ph DMF O

OH OH

1 The X-ray structure of molecular (I) indicates that both 3-hydroxy and 4-hydroxybenzyl groups lie on the same side of five membered ring plane.

Acknowledgment. This work was supported by integration grant SB RAS № 90 (2009).

1 Lozhkin A.V., Sakanjan E.I. Khim.-Pharm. Zhur. 2006, 40, 6, 47.

2 Cheng G. and Hu Y. J. Org. Chem. 2008, 73, 4732.

3 Lee Y.R., Kim B.S. Tetrahedron Letters 1997, 38, 2095.

4 Risitano F., Grassi G., Foti F., Bilardo C. Tetrahedron Letters 2001, 42, 3503.

111 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Alkaloids and Flavonoids from Roots of Sophora flavescens Soland. Isolation and Identification

Natalia A. Pankrushina1,2, Vladimir G. Vasiljev1, Elena I. Chernyak1, Denis V. Sandanov3, Vladimir V. Boldyrev2,4, Byung-Hun Um5

1Novosibirsk Institute of Organic Chemistry SB RAS, Lavrentjev Ave., 9, 630090, Novosibirsk, Russia, E-mail: [email protected] 2Novosibirsk State University, REC 008, Pirogova 2, 630090, Novosibirsk, Russia 3 Institute of General and Experimental Biology SB RAS, Sakhyanovoi str., 6, 670047, Ulan-Ude, Russia 4 Institute of Solid State Chemistry and Mechanochemistry, Kutateladze 18, 630128, Novosibirsk, Russia 5Korea Institute of Science and Technology, Gangneung, Republic of Korea

Natural biologically active substances obtained from medicinal plants are known to be the basis for many remedies. Routine technology of their isolation is the liquid extraction process. This technology requires high consumption of toxic organic solvents produces detrimental waste and becomes environmentally unsafe. So our purpose was to search new effective methods of natural product isolation from plant material. We have suggested the new method of “dry” mechanochemical isolation based on combined high-energetic mechanical treatment of plant material with solid substances of the appropriate chemical nature.1 Applying the technique we use planetary and vibration accelerated grinders. High effectiveness of new mechanochemical approach is proved by isolation of active quinolizidine alkaloids and flavonoids from Sophora flavescens roots with higher yield in compared with a conventional liquid extraction. Identification of alkaloids was performed using 1H, 13C NMR, CMS, HPLC and X-ray methods. Identification of flavonoids was carried out with the help of HPLC and HPLC-MS methods using standard pure samples.

DAD1 D, Sig=370,60 Ref=850,50 (D:\DATA2008\LC 1200\PANKRUSH INA\SF-CLEX_1N.D) mAU H H H 18.736 Kuraridin HO OH HO OH 500 HO O HO OH HO O OH

400 O O O O OH O Kurarinon Kuraridin Kushenol A

300

H H Sophoraflavanone G Sophoraflavanone OH HO OH OH 200 Kurarinone HO O HO O HO O 17.560 17.326

Leachianone A Leachianone O 16.594 100 Isoxanthohumol 15.824 16.837 A Kushenol OH O OH O O O 17.202 18.193 18.369 18.572 16.253 16.967 19.720 15.673 19.919 17.913 Leachianone A Sophoraflavonone G Isoxanthohumol 16.118 19.355 19.544 0 15.5 16 16.5 17 17.5 18 18.5 19 19.5 min

HPLC-CMS profile for CHCl3 extract of the Sophora flavescens roots. ES neg./pos. scan 100-1000, 0.2 ml/min, N2-7 l/min, 220 C, Nb-2.0 XDB-C8, 2%HCOOH-ACN (20-90%, 5- 15'), 0.2 ml/min The financial support of this study from the U.S. Civilian Research & Development Foundation, CRDF (Award No. RUX0-008-NO-X1) is gratefully acknowledged.

1 Pankrushina, N. et al. (2001) Patent RU N 2176919 C2., Inventions, useful models, 35, N 1: 159.

112 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Novel Methods of Mechano-Enzymatic Modification and Recovery of Agriculture Waste

A.A. Politov, O.V. Golyazimova

Novosibirsk State University, Research and Educated center “Molecular Design and Ecological Safe Technology”, Pirogova, 2, Novosibirsk Institute of Solid State Chemistry and Mechanochemistry, Kutateladze, 18, Novosibirsk, 630128, Russia e-mail: [email protected]

Comminution of raw plant materials is a major technological procedure that renders possible a lot of wholesale manufactures, such as paper and woodworking industries, processing of wood flour, feed and food production. Comminition procedure is also used for recovery agriculture waste and production a number of products and semi-product for pharmacology, oil and chemical industries, alternate energy, new materials and consumer goods production. Large expenditure of energy for grinding required obstructs developing of waste recovery. Preliminary chemical treatment of plants and agriculture waste essentially facilitates following comminution but this method does not ecological friendly and is not acceptable feed or food production as well as pharmacology. On basis of conception of polymer strength a novel method of lignocellulose materials destruction was developed in the in Institute of Solid State Chemistry and Mechanochemistry of SB RAS. This new approach combines biotechnological and mechanical treatments of raw materials that results in increase of small particles yield. The increase of grinding efficiency of agriculture waste or plants achieves without additional mechanical energy. For example, using new technology makes twice as much a mass of particles less than 80 micron (see Fig.1). This result is obtained at the same expenditure of energy. On the other hand production of wood or plant flour with needed size composition takes now significantly less energy. Soft communition of raw plants provides a good quality of feed and does not destroy bioactive substances. Using of bio technology is shown to revive such old challenges as bioethanol production from lignocellulose raw materials, ecologically friendly bioplastics, expands nutritive base and develops other new green technologies.

40 a b 80

30 60

20 40

10 20 Mass fraction, % fraction, Mass Mass fraction, % fraction, Mass

0 80-125 200-300 400-500 80-125 200-300 400-500

Particle size,  Particle size, 

Fig.1. Size distribution of rice husk grinded with the same mechanical energy: a – ordinary grinding, b – grinding with bio technology treatment.

This work was supported by ISTC № 3235.

113 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Unique Siberian Soybean Cultivar SibNIIK-315 and Tendency of Plant Breeding

Nikolay I. Kashevarov, Revmira I. Polyudina, Dmitry A. Potapov, Olga A. Rozhanskaya

Siberian Research Institute of Fodder Crops, Krasnoobsk, Novosibirsk Region, Russia 630501 E-mail: [email protected]

Soybean (Glycine max (L.) Merr.) is most important high protein and oil-yielding crop in the world. Sown area of soybean in Siberia is increased as a result of breeding of new Siberian cultivars. Unique high adaptive and early maturing cultivar SibNIIK-315 is widespread and needed in Russia and Kazakhstan. It was developed by V.E. Gorin and colleagues in 1991 by method of individual selection from Swedish accession (k-5828) of VIR genetic collection. The vegetation period of SibNIIK-315 is 90–110 days, seed yield up to 3 t/ha, seeds contain 35-40% protein, and 17-20% oil. The hybridization, mutagenesis, and somaclonal variations in combination with selections are used in Siberian Research Institute of Fodder Crops as the methods of development of new breeding material with high variability of characters (Fig.). Somaclonal variations made it possible to recovery native genetic diversity of ancient cultivated plants of G. max.

120

100

80

60

40

20

0 SibNII 1 R-09 2 R-09 3 R-09 4 R-09 5R-09 6R-09 K-315

somaclones

height up to first bean, cm number of seeds per plant maturing, days

Fig. Characters of new breeding material of soybean

Currently, genetically modified soya is prevalent ingredient in production staple foods. Therefore absence of genetically modified organisms among Siberian cultivars is especially important for consumer.

114 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Effects of Hybrid Macromolecular Phenolic Antioxidants on Whole Blood Viscosity

Elena V. Popova1, Mark B. Plotnikov1*, Oleg I. Aliev1, Nina S. Domnina2, Olga V. Sergeeva2

1 Institute of Pharmacology, Siberian Department, Russian Academy of Medical Sciences, Lenin street, 3, 634028, Tomsk, Russia Fax: 8(3822)41-42-35 E-mail: mbp2001@ mail.ru 2 St. Petersburg State University, Russia

At the present time, development of functional polymer-based nanomedicines is considered to be one of the most advanced trends in medicine. Advantages of these nanomedicines are obvious: an increased efficacy of the medical drug and possible precise drug delivery under the protective system, solubility adjustment and toxicity reduction. It is known that blood viscosity may correspond to the status of health or disease. For example, hyperviscosity syndrome has been observed in patients with cardiovascular diseases, endocrine disorders, neoplastic diseases and intoxication. In this respect the detection of polymer-based nanomedicines, which are able to reduce hyperviscosity, has become a matter of high priority. The objective of this study was to research the hemorheologic activity of hybrid macromolecular phenolic antioxidants (HMPAs) using the model of blood hyperviscosity ex vivo. Structure of HMPAs consist of sterically hindered phenol (potassium phenosan (PP)) and different polymers: dextran (40 kDa) (D), hydroxyethylated starch (200 kDa) (HES) and polyethyleneglycol (20 kDa) (PEG). HMPAs were intravenously administered in doses of 10 mg/kg. Experiments were carried in Wistar rats. The viscosity of whole blood was registered by the rotary viscometer AKR-2 at shear rates of 3–300 sec–1 immediately after blood sampling and in 60 min after blood sample incubation at 20.0±0.4oC (the model of blood hyperviscosity). The intravenous administration of HES-PP reduced hyperviscosity levels by 5–12% before and after blood incubation at shear rates 50, 100, 300 sec–1 compared to the control group (polymer-administered only). The administration of PEG-PP resulted in limitation of hyperviscosity formation (by 6–7%) before incubation at low shear rates, and at the end of incubation process (by 7–11%) in a wide shear rate range (5–300 sec–1). When D-PP administered, hyperviscosity levels were reduced by 6–20% after blood sample incubation at shear rates 3–100 sec–1. Thus, HMPAs can be considered as perspective polymer-based nanomedicines for correction of hemorheologic disorders.

115 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Ecdysteroids of Serratula quinquefolia Bieb. ex Willd

Yana I. Pylina1, Svetlana O. Volodina1, Dmitry S. Bacharov2, Vladimir V. Volodin1

1Institute of biology of the Komi Sci Centre, Ural Division, Russian Academy of Science, 28 Kommunisticheskaya Str., 167982 Syktyvkar, Russia, E-mail: [email protected] 2Syktyvkar State University, Oktyabrsky avenue, 167001 Syktyvkar, Russia

At present it is well known about the influence of the factors of the environment on the qualitative and quantitative composition of biologically active substances in the medicinal plants as a result of biochemical adaptation. We carried out a study of the composition of ecdysteroids in the plants of Serratula quinquefolia Bieb. ex Willd. by their introduction into the conditions of the European northeast of Russia (taiga zone of the Republic of Komi). It is established that the leaves of cultivated plants contain not 20-hydroxyecdysone, specific for the wild plants, growing in the mountain forests of the North Caucasus, but its structural isomer – 25S-inokosterone, whose structure is proven by the methods of mass spectrometry and NMR-spectroscopy. To understand the reasons for this phenomenon we have studied the physiological reactions of the both wild and cultivated plants. It is established that the plastid apparatus of plants in the natural populations (under forest canopy) is adapted to the conditions for the moderate illumination. The suppression of the processes of photosynthesis without the destruction of photosynthetic pigments occurs during the introduction of this plant species into the conditions of long light day on the European northeast of Russia. Switching the biosynthesis of ecdysteroids from 20-hydroxyecdysone to its structural isomer of 25S- inokosterone in the leaves, but not in other parts of the plants (leafstalk, stem, root and rhizome) testifies for the connection between the biosynthesis of ecdysteroids and the adaptive reaction of plants in response to the damaging action of light. The fact discovered can have practical value for the obtaining of 25S-inokosterone for medicine due to its strong antioxidant and anabolic action by the cultivation of Serratula quinquefolia on the open plots under conditions of long light day in the North.

116 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Working out Optimal Technology for the Substance Isolation from Cuscuta campestris Yuncker

Aizhan I. Zhusupova*, Saniya N. Rakhadilova

al-Farabi Kazakh National University, al-Farabi av. 71, 050038, Almaty, Kazakhstan E-mail: [email protected]

As was proven by the alternative medicine, some dodder species like C. europaea L. and C. chinensis Lam. can be used against different plant, animal and human pathologies. The most wide spread variety in Kazakhstan is the field dodder (C. campestris Yuncker), which is almost unstudied from this point. The aim of the current study was to elaborate an optimal technology for the isolation of substance from the field dodder for its further phytochemical study and assay of possible biological activities. Plant material was gathered in September, 2009 in the Almaty region. It was established that humidity of plant material made up 2.57 %, total ash – 5.40%, ashes, insoluble in 10% HCl – 14.91%, sulphated ashes – 8.26%. All esteemed parameters indicate qualitativeness of raw material and low content of mineral substances in it. Different solvents were tested to find the most suitable one: ethanol and acetone, their 40%, 50%, 70% water solutions. The other factors were restricted as follows: plant material: solvent ratio ranged from 1:8 to 1:10, one extraction lasted for 2 to 48 hours, temperature was in between 22 and 30 0C, and the whole process was repeated 1 to 3 times. By varying all mentioned parameters optimal technological scheme for the isolation of substance in form of dry extract was elaborated, herewith, qualitative value of the obtained substance was used as the principal elective criteria. Eventually, optimal technological scheme was found to be: grounded to 3 mm particle size plant material was extracted with 50% ethanol in ratio of 1:8 w/v, extraction lasted for 10 hours under room temperature of 20-23 0C with double repetition of the whole process of extraction. Joint extract was filtered and concentrated to get the substance in form of dry extract, which may be used for further phytochemical study and assay of possible biological activities.

117 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation

Synthesis of N-(12-Cytisinylethyl)-1-(4-methoxyphenyl)-6,7- dimethoxy-1,2,3,4-tetrahydroisoquinoline and Computer Prediction of Its Biological Activity (PASS)

Sh.B. Rakhimov, V.I. Vinogradova

S. Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences, M. U1ugbek Str., 77, 100170 Tashkent, Republic of Uzbekistan Fax: (998 71) 120 64 75 E-mail: [email protected]

Isoquinoline alkaloids, in particular substituted derivatives of 1-phenyl-1,2,3,4- tetrahydroisoquinoline and cytisine (1), possess different pharmacological action. It was interesting to obtain a range of bimolecular compounds on the base of above substances and to study their biological activities. 1-(4-Methoxyphenyl)-6,7-dimetoxy-1,2,3,4- tetrahydroisoquinoline (2), obtained previously, was used for the synthesis of bimolecular compound 3. The reaction was carried out in dioxan at room temperature using cytisine excess as HCl acceptor.

5 4 H3CO H3CO 6 NH 3 2 12'' 7 5'' + N CH2CH2Cl N CH2CH2 N H CO H CO 1 8'' N 3 3 8 1' N O 6' 2' 10'' 2'' 5' 3' O 4' OCH3 OCH3

1 2 3

The structure of compound 3 was confirmed by 1H NMR spectral data. According to the computer prediction PASS [http://www.ibmh.msk.su/PASS], substance 3 possesses the following activities: Activity Prediction Pa Pi for Activity: 0,742 0,011 Antidyskinetic 0,641 0,018 Hypertensive 0,612 0,009 Analeptic 0,611 0,009 Respiratory analeptic 0,537 0,005 Acetylcholine nicotinic agonist 0,509 0,004 Acetylcholine antagonist

118 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Chemical Study of Artemisia succulenta Ldb.

Bibigul B. Rakhimova, Zhanar R. Shaimerdenova, Sergazy M. Adekenov

JSC “International research and production holding “Phytochemistry”, 100009, Karaganda, Kazakhstan Fax: (7212) 43-37-73 E-mail: [email protected]

Genus Artemisia L. is a one of largest and the most young in philogenetical attitude and perfect genus of Asteraceae family. It is counted more than 500 species, including about 200 species in CIS, 57 species in West Europe, 30 species in North America. Representatives of this genera is widely widen (spreading) in North hemisphere. They are dominants of plant associations into arid and subarid zones. 81 species of Artemisia are growing on the Kazakhstan territory.1 Artemisia succulenta Ldb. is annual or biennial endemic plant of Kazakhstan flora. It is growing in steppe and desert zones, on salt-marshes and less salt-marshal meadows, on banks of rivers and shores of salt lakes as weed in irrigational regions.1 Chloroform extract of aerial part of Artemisia succulenta, gathered in Betpakdala desert in territory of Aktogai district of Karaganda region in phase of beginning of budation in 2008 year, two crystallic substances is isolated by methods of column chromatography. First substance (1) with m.p. 110-112 0C (petrol ether-ethylacetate) and yield of 0.005 % (calculation on air-dried raw material) is isolated at elution of chromatograthic column with silicagel by petrol ether-ethylacetate (83:17). The IR-spectrum (KBr, ν, sm–1) of (1) exhibited absorbtion bands typical of carbonyl group of γ-lactone (1782 sm–1), carbonyl of ester group (1763 and 1709 sm–1) and links –C-H (2985, 2950, 2916 and 2891 sm–1). Compound (1) is sesquiterpene lactone to physico-chemical data. 0 Colourless crystalles (2) of composition C15H18O3 with point of melting 166-169 C (petrol ether-ethylacetate) and yield of 0.033 % (calculation on air-dried raw material) at elution of column by petrol ether-ethylacetate (7:3). In IR-spectrum (KBr, ν, sm–1) of (2) presented absorbtion bands typical of carbonyl group of γ-lactone (1785 sm–1), connections – C-H (2934 sm–1) and –C=C (1657, 1629 and 1611 sm–1). In UV-spectrum (EtOH) of (2) presented maximum of absorbtion at 240 nm, corresponding to crossing in ,-unsaturated chromophore of enone system. Compound (2) is identified with sesquiterpene lactone of eudesmane type -santonin on the basis of IR-, UV-spectra data, element analysis, melting point and HPLC analysis data.

1 Flora of Kazakhstan. / Edited by N.V. Pavlov – Alma-Ata: Nauka, 1966. – V.IX. – 639 p.

119 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Antimicrobic Activity in vitro of the Extracts Containing Phytoecdysteroids

I.D. Bobaeva, A.M.-T. Bektimirovb, I.M. Boymatova, N.Sh. Ramazanova

a) Institute of the Chemistry of Plant Substances AS RUz, Tashkent b) Scientific research institute Epidemiology, microbiology and infectious diseases Ministry of Health RUz

Plant steroids are of great importance for practice. Some of them are capable to regulate directly the major physiological processes in living organism, others are initial products for manufacture of effective medical drugs. Antimicrobic activity of plant extracts: Silene viridiflora – methanol extract; Ajuga turkestanica – methanol extract; Rhaponticum integrifolium – buthanol extract; Serratula Sogdiana – buthanol extract; Silene viridiflora – buthanol extract had been studied. Aerial parts of plants were extracted by methanol at room temperature. Evaporated extracts were diluted with the distilled water and processed by buthanol. Buthanol extracts were evaporated. The obtained resin dissolved by 96° ethanol. Antimicrobic activity were studied by modified «cell method» (Navashin S.V., 1983) with use of test microorganisms cultures. The account of results carried out visually – on size of inhibition zone of microorganisms growth around cells. Plant extracts possessed by unequal activity concerning a set the test-strains of microorganisms. The expressed antimicrobic action in the relation to gram-positive bacteria. S. aureus and B. cereus was indicated. Moderate antimicrobic action is noted in the relation of Candida krusei. Ingibition of gram-negative bacteria E.coli and Ps.aeruginosa was not observed, i.e. these groups of microorganisms were resistant to investigated extracts. The studied plant extracts possess the expressed antimicrobic action in the relation to gram-positive bacteria and moderated – in the relation to Candida. The highest antimicrobic activity is established in three plant extracts: Silene viridiflora – methanol extract; Rhaponticum integrifolium – buthanol extract; Silene viridiflora – buthanol extract.

120 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Two Major Stilbene Glycosides of Pinus sibirica Bark: Isolation and Chemical Transformation

Alexandra V. Pozdeeva, Nina I. Komarova, Artem D. Rogachev, Dina V. Korchagina, Nariman F. Salakhutdinov, and Genrikh A. Tolstikov

Vorozhtsov Novosibirsk Institute of Organic Chemistry, Akad. Lavrentiev ave. 9, Novosibirsk 630090, Russia Fax: +7 383 330-88-70 E-mail: [email protected]

Stilbenes are a class of biologically active compounds that have been shown to possess various medicinal properties1. It was found2 that the bark of Pinus sibirica contained two major stilbene glycosides – resveratroloside (1) and pinostilbenoside (2). In our work we have determined the content of both glycosides which appeared to be 1.1-1.3% for glucoside 1 and 1.5-1.8% of glucoside 2 calculated on dry bark weight. For chemical transformations of sugar moiety OGlu being possible, we have synthesized R1O dimethoxyderivative 3 in several ways. When treated 1 R1 = R2 = H 4 R1 = R2 = Tr by CH3I with K2CO3 in boiling acetone, 2 R1 = H, R2 = CH3 5 R1 = Tr, R2 = CH3 3 R1 = R2 = CH3 OR2 resveratroloside 1 hasn’t undergone any changes Glu = -D-glucopyranoside probably because of its low solubility. When solvent was changed on CH3CN the reaction had ended in 3 hours in the same conditions. Methylation of 1 by Me2SO4 in dry CH3CN with

NaOH is not observed. When 10% vol. of H2O is added in this system, the formation of monomethyl derivative 2 with insignificant quantity of 3 occurs. Increasing of water concentration up to 40% vol. leads to formation of compound 3, methylaton of sugar moiety in this case being not observed. Attempts to methylate the compound 1 selectively on aromatic hydroxyls by Me2SO4 with NaOH in the mixtures of EtOH and H2O with different concentrations of water were unsuccessful. In this system methylation of at least one sugar hydroxyl takes place.

When glycoside 1 interacts with TrCl in DMF in the presence of K2CO3 there is no trytilation of the substrate, in particular on CH2OH-group of sugar residue. Treatment of glycosides 1 and 2 by TrCl with NEt3 and catalytic amounts of DMAP in dry MeCN leads to the formation of compounds 4 and 5 respectively after stirring of reaction mixture for 24 hours at room temperature, the sugar moiety in this reaction being unchanged. All reactions were controlled by HPLC with ultraviolet detection in multiwave mode. Structures of all compounds were determined using NMR 1H and 13C spectroscopy.

1. Resveratrol in health and disease, eds. Bharat B. Aggarwal and Shishir Shishodia, CRC Press, Taylor & Francis Group, Boca Raton-London-New York, 2006, 684 p. 2. Gromova, A. S.; Tyukavkina, N. A.; Lutskii, V. I.; Kalabin, G. A.; Kushnarev, D. F. Chem. Nat. Comp. 1975, 11, 715-719; b) Shul'ts, E. E.; Petrova, T. N.; Komarova, N. I.; Salakhutdinov, N. F.; Tolstikov, G. A. Russ. Pat. RU 2294919.

121 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Distribution of Enantiomers of Monoterpene Hydrocarbons in Siberian Plants of the Apiaceae (Umbelliferae) Family

Elena P. Romanenkoa, Elena V. Karpovaa,b, Elena A. Korolyukc, Alexey V. Tkacheva,b

a Novosibirsk Institute of Organic Chemistry, 630090, Novosibirsk, Russia b Department of Natural Sciences, Novosibirsk State University, Pirogova str., 2, 630090, Novosibirsk, Russia c Central Siberian Botanical Garden, Zolotodolinskaya Str. 101, 630090 Novosibrisk, Russia Fax: +7 (393) 330-97-52 E-mail: [email protected]

In many countries people have used numerous plants of the Apiaceae family for medicinal and flavoring purposes and the green parts as vegetables for hundreds of years. There are dozens of papers on chemical composition of the Apiaceae volatiles, but little is known on enantiomeric composition of the main constituents except for several commercially available oils and some other species. We describe now the enantiomeric composition of monoterpene hydrocarbons in essential oils of plants of the Apiaceae family growing wild in Western Siberia: Angelica decurrens (Ledeb.) B. Fedsch., Aulacospermum anomalum Ledeb., Bupleurum bicaule Helm, Bupleurum multinerve DC., Bupleurum scorzonerifolium Willd., Carum carvi L., Cenolophium denudatum (Hornem.) Tutin, Chaerophyllum prescottii DC., Cicuta virosa L., Ferulopsis hystrix (Bunge) Pimenov, Heracleum dissectum Ledeb., Heracleum sibiricum L., Kadenia dubia (Schkuhr) Lavrova et V.N.Tichom., Kitagawia baicalensis (Redow. ex Willd.) Pimenov [Peucedanum baicalense (Redow. ex Willd.) W.Koch], Pastinaca sylvestris Miller, Peucedanum morisonii Bess. ex Schult., Peucedanum vaginatum Ledeb., Pimpinella saxifraga L., Seseli buchtormense (Fischer ex Sprengel) W. Koch, Seseli condensatum (L.) Reichenber. fil., Seseli ledebourii G. Don fil., Seseli libanotis (L.) Koch [Libanotis sibirica (L.) C. A. Mey.], Sium latifolium L., Sium sisaroideum DC., Sphallerocarpus gracilis (Besser ex Trev.) Koso-Pol. [Chaerophyllum gracile Besser ex Trev.] where collected from natural habitat. Essential oils were prepared by steam distillation of the fresh grass in the field-distillation device made of stainless steel and studied by enantioselective GC-MS using an Agilent 6890/5973N EI/PCI instrument equipped with a CycloSil-B chiral capillary column.

This work was supported by Russian Foundation for Basic research (Grant # 07-03-00620-a).

122 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis of New Lappaconitine Derivatives Containing Indolizine Framework

Vasiliy E. Romanov, Elvira E. Shults, Genrikh A. Tolstikov Novosibirsk Institute of Organic Chemistry, SB RAS, Novosibirsk, Russia, e-mail: [email protected]

The diterpene alkaloid lappaconitine (1) isolated from plants of the Aconitum genus is used as hydrobromide salt as an antiarrhythmic remedy [1]. However, the high toxicity of this alkaloids consider the synthetic transformation very importance. Indolizine framework has been an interesting heterocyclic moiety because of the presence of this skeleton in a number of bioactive natural products [2]. Several indolizine derivatives, in fact, have been known to exhibit various biological activities such as antioxidant [3], calcium entry blockers [4], 5- hydroxytriptamine (5-HT3) receptor antagonists [5] and antileukemic activity [6]. We describe the synthesis of compounds containing lappaconitine and indolizine moieties from 5'-iodolappaconitine (2) [7]. The cross-coupling reaction of 5'-iodolappaconitine (2) with 5- vinyl-2-methylpyridine produced styrylpyridine derivative (3) (yield 72%) (Scheme). Pyridinium salts (4–6) prepared by the reaction of compound (3) with phenacyl bromide, 4- methoxyphenacyl bromide, or 3,4-dichlorophenacyl bromide were subjected to 1,3-dipolar cycloaddition with dimethyl acetylenedicarboxylate in the presence of triethylamine. The reaction leads to indolizine derivatives (7–9) in 50–55% yield.

OCH3 OCH3 OCH3 H3CO OCH3 OCH3 HO OCH3 HO OCH3 HO OCH3 N N 1) 10% H SO N 2 4 N OH OH OH 2) ICl, AcOH Pd(dba) O O O 2 O 3) Ac2O P(o-Tol)3, O Br O C O C C Et3N, DMF H Ar 2' 1' 5' C NHAc AcHN AcHN I H3C C 1 N 2 H 3

OCH3 OCH3 H CO HO OCH3 3 OCH HO 3 OCH3 CO Me N N 2 H3C OH OH O O + O H N H CO Me H C C O 2 3 C C O C C C Et N O N 3 H R2 H NHAc - NHAc CO2CH3 Br 1 R1 R 4-6 H3CO2C R2 7-9 1 2 1 2 1 2 R =R =H (4, 7); R =OCH3, R =H (5, 8); R =R =Cl (6, 9).

1 Mashkovskii, M.D., Lekarstvennye sredstva (Drugs), Moscow: Novaya Volna, 2006. 2 Lotter, A.N.C.; Pathak, R.; Sello, T.S.; Fernandes, M.A.; Otterlo, W.A.L.;, de Koning, C.B. Tetrahedron, 2007, 63, 2263. 3 Ostby, O.B.; Dalhus B., Gundersen L.-L.; Rise, F., Bast, A.; Haenen, G.R.M.M. Eur. J. Org. Chem., 2000, 3763. 4 Gubin, J.; Vogelaer, H.D.; Inion, H.; Houben, C.; Lucchetti, J.; Mahaux J.; Rosseels, G.; Peiren, M.; Clinet, M.; Polster, P.; Chatelain, P. J. Med. Chem., 1993, 36, 1425. 5 Bermudez, J.; Fake, C.S.; Joiner, G.F.; Joiner, K.A.; King, F.D.; Miner, W.D.; Sanger, G.J. J. Med. Chem., 1990, 33, 1924. 6 Anderson, W.K.; Heider, A.R.; Raju, N.; Yucht, G.A. J. Med. Chem., 1988, 31, 2097. 7 Osadchii, S.A., Shul’ts, E.E., Polukhina, E.V., Shakirov, M.M., Tolstikov, G.A. Rus. Chem. Bull., Int. Ed. 2006, 55, 1077.

123 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Testing Cytokinin Activity of Rice Husk Mechanochemical Preparation with the Help of Culture Chickpea (Cicer arietinum L.) in vitro

Olga A. Rozhanskaya1, Oleg I. Lomovsky2

1 Siberian Scientific Institute of Fodders SB RAAS, Krasnoobsk Novosibirsk region, 630500, Russia 2 Institute of Solid State Chemistry and Mechanochemistry SB RAS, ul. Kutateladze 18, Novosibirsk, 630128, Russia E-mail: [email protected]

The regeneration of whole plant organisms from partial somatic cells or from isolated tissues is a key moment in agricultural planting and selection. The special phytohormones – cytokinins are introduced into microbiological feeding matrix to stimulate cell growth and to induce regeneration. The goal of work is the testing of activity of preparations produced from rice husk via mechanochemical methods. The preparations were synthesized by mechanical treatment of rice husk containing mixtures under controlled intensity and duration. Explants of chickpea seeds are placed on feeding matrix Gamborg B5 containing produced preparations as growth regulators. Preparations are demonstrated the high cytokinin activity during 3 weeks excelling action of synthetic cytokinin 6-BAP (6-benzylaminopurine) in control experiment. The regeneration frequency is increased 4%, risogenesis 25%. The growth of shoots, leaves and roots was accelerated. Callus formation frequency was decreased in factor 2.

Figure. Influence of preparation upon morphogenesis in culture of chickpea tissue (incubation time 3 weeks): 1 – regeneration frequency,%, 2 – shoot height, mm, 3 – rysogenesis frequency, 4 – root length, 5 – callus genesis frequency. White columns – B5 + 6-BAP 0.5 mg/l, black columns – B5 + rice husk preparation 1.0 mg/l.

So, the experimental results how s the possibility of utilization of rice husk mechanochemical preparation as an effective morphogenesis regulator in biotechnology instead of synthetic citokinin and the perspectives of rice husk preparation application as growth stimulator for planting.

124 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Cathepsin L–Like Protease from Dermestid Beetle Dermestes frischii Maggot

G.N. Rudenskaya a, A.S. Javadov a, Yu.A. Rudenskaya b, A.I. Papisova a, G.N. Balandina a, R.D. Zhantiev b O.S. Korsunovskaia b, Y.E. Dunaevsky c

a Department of Chemistry, Moscow State University, Moscow 119992, Russia b Department of Biology, Moscow State University, Moscow 119992, Russia c A.N. Belozersky Institute of Physico-Chemical Biology, Moscow 119992, Russia Fax: +7(495) 9393181 E-mail: [email protected]

A cathepsin L-like protease from dermestid beetle Dermestes frischii maggot guts was obtained and investigated. Protease was isolated through affinity column at arginine-diasorb followed by FPLC gel-filtration at Superdex G-75. Protease is active against chromogenic peptide substrates, containing Arg in P1 position and a hydrophobic residue in P2 position. Collagenolytic activity was identified by hydrolysis of azocollagen at 37? C. PH optimum is about 8,0 and temperature optimum at 58?C. Enzyme is inhibited completely by HgCl2 and leupeptin that prove it’s belonging to cysteine proteases of papain family. cDNA analysis of cathepsin L-like protease showed that protein sequence consists of 339 amino acid residues. Mature cysteine protease contains 219 mino a acid residues corresponding to molecular weight 24027,20 Da. Residues of the active site were identified: Gln140, Cys146, His285, Asn306 and Trp308. Calculated pI is 4,73. Cysteine protease’s amino acid sequence from dermestid beetle shows high structure homology with cathepsin L of other insects.

125 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Synthesis and Spatial Structure of 4-(2-Isothiocyanatopropan-2-yl)methyl-1-thiocyanatocyclohexane

Daniyar T. Sadyrbekov1, Oleg G. Ryazantsev1, Elena V. Tikhonova1, Almagul Mahmutova1, Koblandy M. Turdybekov1, Yurii V. Gatilov2, Gayane A. Atazhanova1, Sergazy M. Adekenov1

1 JSC “International research and production holding “Phytochemistry”, 100009, Karaganda, Kazakhstan Fax: (7212) 43-37-73 E-mail: [email protected] 2 N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, SB RAS, Novosibirsk, Russia

At interaction of limonene (1) with crystal thiocyanate sodium and the subsequent o addition of 73% H2SO4 at heating up to 85 C derivatives (2) is synthesized with the yield of 7%.

SCN

H2SO4, NaSCN

NCS

(1) (2) The spatial structure of molecule (2), proved by method X-ray structural analysis, is presented in figure:

Thus, as a result of this work, the derivative of limonene was obtained for the first time, having structure 4-(2-isothiocyanatopropan-2-yl)-1-methyl-1-thiocyanatocyclohexane and its spatial structure was determined.

126 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Dependence of N-Desacetyllappaconitine Hydrobromide from the Method of Its Preparation

Sh.Sh. Sagdullaev, A.Z. Sadikov, N.V. Valiev

Acad. S.Yu. Yunusov Institute of chemistry of plant substances AS RUz 77, M.Ulugbek street, Tashkent, Uzbekistan, phone (998-71) 262-71-00, fax: (998-71) 262-73-48, email: [email protected]

Earlier we reported that we had had developed technology of production of preparation N-Desacetyllappacinitine hydrobromide. Our studies showed that alkaloid N- Desacetyllappaconitine is contained in insignificant quantities in plants, mainly it occurs during the production process of medicinal preparation Allapinine from plant raw material. We experimentally proved that N-Desacetyllappaconitine is the metabolite of alkaloid lappaconitine and gets formed at thermal processing of extracts and solutions of bases and various salts of alkaloid lappaconitine (sulphate and bromhydrate of lappaconitine). Water solutions of preparation N-Desacetyllappaconitine hydrobromide from samples of substances obtained initially in the form of base of N-Desacetyllappaconitine from wastes of allapinine production from consequent obtaining of bromhydrate and chlorhydrate salts (N- Desacetyllappaconitine hydrobromide or hydrochloride), turned out to be inconsistent, because new impurities occur in the form of bromhydrate and chlorhydrate salts while keeping water solution of preparation and they significantly alter the color of preparation. Continuing studies on development of industrial technology of obtaining N- Desacetyllappaconitine hydrobromide from waste of production of allapinine from rhizomes and roots of aconitum septentrionale, we got consistent samples of preparation N- Desacetyllappaconitine hydrobromide from master batches of solutions from allapinine production after its re-crystallisation in native form. Thus, it can be concluded that for obtaining consistent samples of N- Desacetyllappaconitine hydrobromide, it is necessary to isolate it from master batches of allapinine solutions only in the form of its bromhydrate salts, without distructing structure of compound formed in the master batch. At the moment, scientific research studies are being conducted on identification of compounds formed during keeping solutions of unstable samples of preparation.

127 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Analytical Control of Production of N-Desacetyllappaconitine Hydrobromide

A.Z. Sadikov, Sh.Sh. Sagdullaev, N.V. Valiev, Sh.A. Otaeva

Acad.S.Yu.Yunusov Institute of chemistry of plant substances AS Ruz 77, M.Ulugbek street, Tashkent,Uzbekistan, phone (998-71) 262-71-00, fax: (998-71) 262-73-48, email: [email protected]

We have developed technology of industrial production of N-Desacetyllappaconitine hydrobromide. Earlier, we reported that we had had developed quantative determination method of substance by the method of spectrophotometer. Temporary pharmacopoeia article has been developed for the substance. Industrial method of production of N-Desacetyllapaconitine hydrobromide mainly consists from the following steps:  obtaining dry residue from solution of master batch, taken after re-crystallisation of allapinine  obtaining water solution of master batch  obtaining chloroform solution of sum of alkaloids  obtaining technical N-Desacetyllappaconitine hydrobromide  obtaining N-Desacetyllappaconitine hydrobromide substance Continuing studies on analytical control of production, we identified main losses and yields of the main product (N-Desacetyllappaconitine hydrobromide) at each technological steps of production. Our studies showed that dry residue of master batch taken after re-crystallisation of allapinine, contains 50% of N-Desacetyllappaconitine hydrobromide, about 35% of lappaconitine hydrobromide and 15% hydrobromide salts of other accompanied alkaloids. Water solution of master batch contain about 90%, technical N-Desacetyllappaconitine hydrobromide contain 95-97% of N-Desacetyllappaconitine hydrobromide, and N- Desacetyllappaconitine hydrobromide substance must contain not less than 97% of main active ingredient. Thus, we have received results of identification of main yields and losses of N- Desacetyllappaconitine at technological production steps.

128 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Chromatographic Method of Purifying Substance of Acsaritmin

A.Z. Sadikov, Sh.Sh. Sagdullaev, Z.U. Mullabaeva

Acad. S.Yu. Yunusov Institute of chemistry ofplant substances AS RUz 77, M.Ulugbek street, Tashkent, Uzbekistan, phone (998-71) 262-71-00, fax: (998-71) 262-73-48, email: [email protected]

A medicinal preparation name Acsaritmin has been created in the institute of chemistry of plant substances, which represents sum of main alkaloids, isolated from rhizomes with roots of Aconitum septentrionale. Works on the development of its industrial method of production are being conducted. At the moment, optimal conditions of extraction of alkaloids from plant raw material, processes of liquid-liquid extraction in homogenic systems for purification from non-alkaloid impurities, and processes of obtaining technical product have been developed and established. Our studies of purification of technical product showed that acsaritmin fabricate from technical product can be prepared with 3 method: 1. purification with activated char coal of sort A, 2. liquid-liquid method in homogenic systems, 3. chromatographic method of purification with the application of various sorbents. Method of chromatographic purification with the application of aluminium oxide with the level of activity V by Brocman has turned out to be rational and economically effective. This method has been worked out on experimental equipment created in the Experimental production of ICPS AS RUz. Necessary quantities of samples from preparation's substance was produced for preparation of important technical documents, method of quality control and preparation of finished drug forms – tablets of acsaritmin 0.025g covered with layer. Necessary quantities of finished drug forms of preparation were prepared for providing clinical trials in cardio logical department of 3 clinics in Uzbekistan.

129 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Spectrophotometric Method of Analysis of Substance of Acsaritmin Preparation

A.Z. Sadikov, Sh.Sh. Sagdullaev, Z.U. Mullabaeva, Sh.A. Otaeva

Acad.S.Yu.Yunusov Institute of chemistry of plant substances AS RUz 77, M.Ulugbek street, Tashkent,Uzbekistan, phone (998-71) 262-71-00, fax: (998-71) 262-73-48, email: [email protected]

Continuing studies on development of drug Acsaritmin, we conducted researches on development of method of quantitative determination of main active compound in plant raw material, intermediate products, technical products and in the substance of acsaritmin for step- by-step control of production of evaluation of substance being obtained. Studying physical- chemical properties of acsaritmin drug, its solubility in various solvents, we offered spectrophotometric method of analysis of this preparation. Our studies showed that acsaritmin drug contains about 9 alkaloids. Preparation contains significant amount of alkaloid lappaconitine which possesses antiarrhythmic activity. Remaining alkaloids from the content of drug possess the same antiarrhythmic activity. We have suggested method of quantification of sum of alkaloids in the substance using the method of titration. Content of lappaconitine the substance of acsaritmin is identified with the method of spectrophotometer. Since, drug allapinine which was earlier developed in ICPS AS RUz contains not less than 96% lappaconitine, allapinine was used as the standard referral sample for quantitative identification of lappaconitine substance in Acsaritmin drug. For identification of quantitative content of lappaconitine, we measure optical density of acsaritmin solution in methanol in spectrophotometer at wave length 310nm in cuvette with the thickness of layer 10mm. We used methanol as a solution for comparison. Simultaneously, we measure optical density of solution of working standard sample of allapinine. Studies of serial samples of drug implied that acsaritmin drug contain not less than 50% lappaconitine and content of sum of alkaloids in the substance is not less than 90%.

130 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation BAS Accumulation and in vitro Propagation of Salix High Productive Genotypes

Roman V. Sergeyev, Nadezhda N. Sheveleva, Alexey I. Shurgin

Mari State Technical University, Lenin sq., 3, 424000, Yoshkar-Ola, Russia Fax: (8362) 646477 E-mail: [email protected]

During last decades there is ncreasing i interest in phytogenous drugs that can be explained by allergic reaction growth to drugs with a synthetic structure. In connection with increase in demand for natural medical products it becomes necessary to create industrial plantations of Salix clonal sorts that are high productive in salycylates. Drugs made from Salix plants ontain c salicin (2-(hydroxymethyl)-phenyl-O- β-D- glucopyranoside) and its derivatives: fragilin, salicortin, 2`-O-acetylsalicortin, tremulacin, salireposide and others.1 There are some problems connected with traditional cutting of certain willow genotypes, for that reason it is necessary to develop in vitro propagation technologies of hybrid willows.2 Willow interpopulation variability in total salicin content was studied. The total salicin content in bark extract of selected genotypes was 7,4 ± 0,2 %. Four different nutrient media were used for the tissue culture establishment: MS by Murashige and Skoog (1962), WPM by Lloyd and McCown (1980), DKW following Driver and Kuniyuki (1984), GD by Gresshoff and Doy (1972). In different experiments the MS, WPM, DKW, GD were supplemented with benzylaminopurine (BAP), kinetin (Kn), or 2- isopentenyl adenine (2-ip) either alone or in combination with naphthaleneacetic acid (NAA). During cultivation of explants from selected populations on MS medium correlation of 0,2 mg/l Kn and 0,01 mg/l NAA caused new shoot formation (4,2 per explant) after four- week cultivation. Root formation was observed on WPM medium after adding 0,2 mg/l NAA.

1 Wagner, H.; Bladt, S. New York: Springer 2001, XVI, 384. 2 Liesebach, M.; Naujoks, G. Plant Cell, Tissue and Organ Culture 2004, 79, 239–247.

131 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Distribution of Saponins in Fabaceae

Dmitry M. Shadrin1, Alexander V. Rodionov2, Svetlana O. Volodina1, Kirill G. Tkachenko2, Vladimir V. Volodin1

1 Institute of Biology, Komi Sci Centre, Ural Division, Russian Academy of Science, 28 Kommunisticheskaya Str., 167982, Syktyvkar, Russia E-mail: [email protected] 2 Komarov Botanical Institute of the Russian Academy of Sciences, 2 Prof. Popova Str., 197376, Saint Petersburg, Russia

Saponins (steroid and triterpenoid glycosides) are the very extended group of the plant secondary metabolites. At the present time it is known more than 200 different structures, which are discovered in more than 70 families of vascular plants. More often such compounds are found in the plants of the families of Fabaceae, Primulaceae, Lamiaceae and Araliaceae. Both triterpenoid and steroid glycosides are widely use in medicine. It is considered that in plants saponins serve as the defence factors in the interactions with other organisms. For example, many triterpenoid saponins and steroid glycosides of a spirostanol type suppress a growth of the phytopathogenic fungi. Furostanol glycosides are toxic for some insects. This work is directed to reveal the regularities between the distribution of saponins of the different types and the phylogenetic classification of plants (on the example of fam. Fabaceae). This approach will make it possible to estimate their significance as the chemotaxonomic markers, and also functional role of saponins in the adaptation and evolutions of plants. For determining the saponins the known express physico-chemical and biological methods of testing the plants were used: capability for foam formation, haemolysis of erythrocytes, and also TLC- and HPLC-chromatography. The methods of mass spectrometry and NMR- spectroscopy were used for establishing the fine structure of saponins. Samples of 39 species from 14 genera were studied for the saponins presence. It was established that the overwhelming majority of the species studied contains triterpenoid glycosides. Steroid glycosides are discovered only in the plants of the genus Trigonella – T. foenum-graecum (tribe Trifolieae) and, supposedly, in Anthyllis vulneraria (tribe Loteae). For explaining the revealed special features of he t distribution of saponins we reconstructed molecular phylogeny of the family Fabaceae. For the analysis of closely-related taxons were used the sequences ITS1 and ITS2 from the data base of the GenBank. The presence of steroid glycosides only in the representative of Trigonella genus in the tribe Trifolieae, but not in other genera of this tribe can be explained by a certain isolation of the genus Trigonella in this tribe that in turn reflects the result of the evolution of this taxon. The regularities of the distribution of saponins among the members of Fabaceae have been used for the development of the chemotaxonomic forecast of their discovering in the flora of the European northeast of Russia in our present studies.

132 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Polyprenols of Cotton Leaves and Their Pharmacological Activity

Kh.M. Shakhidoyatov, N.K. Khidyrova, N.M. Mamatkulova, V.N. Syrov, Z.A. Khushbaktova

Institute of the Chemistry of Plant Substances, Academy of Sciences, Republic of Uzbekistan, Mirzo Ulugbek str., 77, Tashkent. Fax: 998-71-120-64-75 E-mail: [email protected]

For the last 25 years the preparations on a basis of plant polyprenols are intensively developed. In this reason searching of plant sources of polyprenols from local plant raw materials is an actual aim. Researches on studying of leaves polyprenols of 12 lines and 3 types of cotton differing among themselves by phenological signs, and also by properties of independent regulation of growth are carried out. It is found out the presence of polyisoprenols such as: polyprenols, bombyprenones, glicynoprenols and diols in the cotton leaves. Dynamics of their accumulation in a plant ontogenesis is investigated and is revealed, that their maximum accumulation is necessary in a phase of maturing of cotton plant. The methods of isolation of polyisoprenoids and biological active additive (BAA), rendering anti-ulcer effect and increasing indicators of lipid metabolism of a skin from cotton leaves are developed.

133 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Isolation of Betulin from Birch Bark by Supercritical Alcohols

Viktor I. Sharypov1, Sergey V. Baryshnikov1, Natalia G. Beregovtsova1, Boris N. Kuznetsov1,2*

1Institute of Chemistry and Chemical Technology SB RAS, K. Marx str.,42, Krasnoyarsk, 660049 2Siberian Federal University, Krasnoyarsk fax: +7(391)2439342, e-mail: [email protected]; [email protected]

The known methods of betulin isolation from birch bark at supercritical conditions use basically the gaseous reagents (CO2, light hydrocarbons). The supercritical extraction makes it possible to decrease considerably the time of betulin isolation and to simplify its separation from extractant. But the serious disadvantage of these methods is the high pressure (20-60 MPa) needed for betulin extraction with a high yield. In the present study the processes of betulin extraction from birch bark by ethanol and isopropanol at subcritical (2 MPa, 100 °C) and supercritical (6.5 MPa, 245 °C) were compared. Experiments were carried out in a flow reactor. It was found that the extraction of birch outer-bark by subcritical alcohols at 100-150 °C increases by 1.5 time the yield of betulin in comparison with extraction at atmospheric pressure. The rate of betulin extraction from bark is significantly increased in supercritical alcohols at 245 °C.

Table. Extraction of birch bark and birch outer-bark by ethanol under the pressure Yield of products, % mas. Raw material Conditions of extraction Betulin Suberin Ethanol, subcritical* 10.9 1.4 Isopropanol, subcritical* 10.3 2.6 Birch bark Ethanol, supercritical** 14.2 4.8 Isopropanol, supercritical** 13.7 5.1 Ethanol, subcritical* 29.2 2.5 Isopropanol, subcritical* 32.2 1.4 Birch outer-bark Ethanol, supercritical** 28.9 8.5 Isopropanol, supercritical** 29.7 9.6 * 150 °C, 2.5 MPa, 3 h; ** 245°C, 6.5 MPa, 0.5 h

By pressure reduction or cooling it is possible to isolate the solid extract with yield up to 75 % from total extract. According to HPLC method, the isolated extract contains near 90 % of betulin and admixtures of lupeol and betulonic acid.

134 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Cryoconservation of Cultures of Cells of Endemic Species of Plants

Z.K. Shaushekov, A.Zh. Kuandykova, A.B. Eshmagambetova, G.K. Asanova, S.M. Adekenov

JSC “International research and production holding “Phytochemistry”, 100009, Karaganda, Kazakhstan Fax: (7212) 43-37-73 E-mail: [email protected]

It is considered an actual conservation of genofond of rare and endemic species of plants in conditions in vitro by means of their introduction in callus cultures and with the subsequent storage in cryoconservation conditions of plant material in liquid nitrogen. Objects of research were endemic species of plants: Ajania fruticulosa (Ldb)Poljak., Artemisia leucodes Schrenk., Tanacetum ulutavicum Tzvel. Cells and tissues of endemic species of plants: Ajania fruticulosa (Ldb)Poljak., Artemisia leucodes Schrenk., Tanacetum ulutavicum Tzvel. for freezing were selected in the middle of exponential phase of the growth curve. Before freezing the callus tissues were preliminary cultivated on nutrient mediums MS, containing various substances – mannitol for reduction of the size of vacuoles, proline for binding of water in a cell, and also it was carried out an artificial hardening to a cold. In experiments three variants of cryoprotectors were used: penetrating cryoprotector glycerin, nonpenetrating cryoprotector saccharose and a mix of these two cryoprotectors. Viability of callus tissues was estimated by vital stains (painting the dead cells). Then the renewal of growth was carried out at cultivation on standard medium MS. In experiments on cryoconservation of callus cultures of endemic species with use of a method of freezing by the best cryoprotector of the mix of glycerin and saccharose. Use of such complex of methods allowed to carry out for the first time the successful cultivation of endemic species of plants. Thus, within the limits of carried out research it s i determined that the method cryoconservation is actual and important for conservation of a genetic variety of endemic species of plants of natural flora.

135 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Step-by-Step Preparation of Spironorketone from 2,3-Seco- 19β,28-epoxy-28-oxo-18α-olean-2,3-dicarboxylic anhydride

Andrew V. Shernyukov, Iliya Ya. Mainagashev, Dina V. Korchagina, Nariman F. Salakhutdinov, Genrikh A. Tolstikov

N.N. Vorozhtsov Novosibirsk Institute of Organic Chemistry of Siberian Branch of Russian Academy of Sciences Lavrentjev Ave. 9, Novosibirsk, 630090, Russia Fax: +7(383)3309752 E-mail:[email protected]

Previously1 we found a new transformation resulting from the reaction of triterpenoid seven-membered cyclic anhydride 1 with benzylamine or p-methoxybenzylamine in THF followed by treatment with oxalyl chloride (“one-pot” conditions). The reaction leads to spironorketones containing a pyrrolidinetrione fragment (compounds like 3). We investigated this reaction step-by-step with phenethylamine. Reaction of compound 1 with phenethylamine in CHCl3 leads to amic acids 2a and 2b, which were isolated by column chromatography over

SiO2 in 57% and 42% consequently. In presence 6-7 fold excess of oxalyl chloride in THF compounds 2a and 2b were converted completely in compounds 3 and 4, which were isolated by column chromatography over SiO2 in 76% and 83% consequently. Thus, desired compounds like 3 could be obtained from 2-amic acids, while 3-amic acids and cyclic imides are not intermediates of present reaction.

O O O H O H O H NH2 HOOC O O 2 O O NH H 3 N CHCl3 HOOC O O 1 2a (57%) 2b (42%)

7x (COCl)2 6x (COCl)2 THF THF

O O N O O H O O O H N O O O

4 (83%) 3 (76%)

1 Shernyukov, A.; Mainagashev I.; Korchagina D.; Gatilov Yu.; Salakhutdinov N.; Tolstikov G. Doklady Chemistry, 2009, 429, 286–289.

136 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Extractive Substances from Twigs of Picea obovata L.

Emma N. Shmidt, Galina F. Chernenko

Novosibirsk Institute of Organic Chemistry, prosp. Lavrent’eva, 9, 630090, Novosibirsk, Russia Fax: +7(383) 330-97-52 E-mail: [email protected]

Wood greens of conifers are widely used for obtaining some biologically valuable products [1, 2]. They consist of needles, twigs, and branches. Green mass of conifers is very good studied [3], but data about extractive substances from twigs are poor. Here we present results of a research of extractive substances from needleless twigs of Picea obovata. Extract, obtained by processing of air-dry raw material with methyl-tret-butyl ether (MTBE), was divided into acid and neutral components, which were later studied separately. Acids (30% from extract) were methylated with diazomethane, methyl esters of the acids were analyzed by method of chromatomass-spectrometry (GC-MS), and individual components were isolated by column chromatography. Acid components were represented by fat acids C16–C32, a sum of resin acids, characteristic for conifers, where dehydroabietic and isopimaric acids prevail. Feature of the extract from twigs is presence of oxidized levopimaric and abietic acids (1-4).

O O O O

COOCH3 COOCH3 1 2

O OH

O O

O

COOCH3 COOCH3 3 4 Structures of isolated compounds are proved by 1H and 13C NMR spectra and RSA.

1 Levin E.D.; Repjah S.M. Pererabotka Drevesnoy Zeleni; Lesnaya Promyshlennost’: Moscow, 1984, 117 p. 2 Vasilyev S.N.; Kushnikova E.A.; Artyomkina N.A. Rastitel’nye Resursy 2001, 4958. 3 Kuchin, A.V.; Chukachova, I.Yu. Abstracts of Papers, Chemistry of Plant compounds and Organic Synthesis, Syktyvkar, June 2-5, 2009; pp 65-69.

137 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Influence of Oxime Pinostrobine on Lipidic Spectrum at Rats with Experimental Diabetes

Leila I. Arystan, Zarina T. Shulgau

JSC “International research and production holding “Phytochemistry”, 100009, Karaganda, Kazakhstan Fax: (7212) 43-37-73 E-mail: [email protected]

The diabetes is the most widespread from endocrine diseases and the important factor of growth of cardiovascular diseases. It is well-known that the diabetes causes the abnormality of an exchange of lipids thereby increasing a risk of development of an early and fast atherosclerosis of vessels of brain, heart and limbs. Many researchers consider the diabetes as a complex exchange-vascular pathology. The purpose of this work is to study the influence of oxime pinostrobine on parameters of lipid spectrum of blood. Oxime pinostrobine is an oxime derivative of flavonoide pinostrobine isolated from gemma of Populus balsamifera L. Experiments were carried out on 22 male-rats in weight 200-250 g. The diabetes, at animals in tested groups, was modeled by intraperitoneal introduction of alloxan in a doze of 90 mg/kg for 4 days. In 7 day after the last introduction of alloxan the maintenance of glucose in blood was determined. Animals with a level of glucose more than 15 mmole/l were taken in experimental groups. A condition of lipid spectrum was estimated on the following parameters: the maintenance in blood of the general cholesterol, triglycerides, high density lipoproteins and low density lipoproteins. The tested animals were injected oxime pinostrobine intragastricaly in a doze of 150 mg/kg for 30 days, the animals received equivolume quantity of 1 % starch mucus in group of the control. The maintenance of the general cholesterol in blood was 53,5±10,3 mg/dl, triglycerides – 37,4±12,0 mg/dl, high density lipoproteins – 12,4±2,6 mg/dl, low density lipoproteins – 33,6±9,4 mg/dl at the intact animals. During the research of lipid spectrum at animals with diabetes the increase in quantity of the general cholesterol was on 67 %, triglycerides – on 33 %, high density lipoproteins – on 89 % and low density lipoproteins – on 73 % is determined. Oxime pinostrobine at rats with an experimental diabetes promoted decrease in the maintenance of oxime pinostrobine on 12 %, triglycerides – on 33 %, low density lipoproteins on 14 %, thus not influencing on quantity of high density lipoproteins. That is, oxime pinostrobine reduced the raised maintenance of cholesterol mainly due to decrease of its atherogenic fraction, not influencing thus on the maintenance of cholesterol in composition of antiatherogenic fraction of high density lipoproteins. Thus, application of oxime pinostrobine at rats with an experimental diabetes allows to correct changes of lipid spectrum of blood.

138 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation The Neutral Compounds of Aerial Parts of Geranium saxatile Kar et Kir.

D.R. Siddikov, N.M. Mamatkulova, S.Z. Nishanbayev, N.K. Khidyrova, Kh.M. Shakhidoyatov

Institute of the Chemistry of Plant Substances, Academy of Sciences, Republic of Uzbekistan, Mirzo Ulugbek str., 77, Tashkent Fax: 998-71-120-64-75 E-mail: [email protected]

Geranium saxatile K. (Geraniaceae) – perennial herbaceous, growing basically in The Central Asia and mountainous regions of Turkmenistan. It’s known that, the aerial parts of Geranium saxatile K. contain steroidal saponins. Other components of that plant have not been researched. On the other hand, in the traditional medicine, an extract of this plant is used as knitting, soft disinfecting and anti-inflammatory preparation, causing interest in studying neutral compounds of Geranium saxatile K. growing on the territory of Republic of Uzbekistan. In the current paper, some information has been provided compounds of aerial parts of plant samples, collected from several regions of Tashkent Province. After consequent extraction with hexane sum of neutral compounds were obtained with the yield 2.2 % and content of ethylacetate and n–butanolic sum was 1.8–2.0 times more. By studying chemical content of neutral compounds, we showed that the extract contains polyisoprenoid alcohols, sterols, higher hydrocarbons and alcohols.

139 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Microcalorimetric Method for Study of Antioxidant Activity of Natural Lipid Fractions

Natalia V. Sizova

Institute of Petroleum Chemistry, SB RAS, 634055, Academichesky ave.3, Tomsk, Russia, Fax: (3822)-491457 E-mail: [email protected]

The search for new effective natural antioxidants (AOX) for the purpose of their application in dietotherapy and cosmetics is now a task of he t great importance. The metabolism suggests radical processes and action of endogenous antioxidants in human organism, but the radical body’s disbalance requires increased concentrations of antioxidants. The preliminary analysis of antioxidant activity of new extracts was carried out through the model reactions. The purpose of this investigation was measurement of AOX concentrations in lipid fractions of plants. Objects of investigation. The following lipid fractions of plants have been studied: natural fatty oils of vegetable origin, essential oils, carbon-dioxide and hydrocarbonic extracts. Method. The measurements have been carried out with the isothermal microcalorimeter MCDP-2, designed and produced in the Institute of Petroleum Chemistry, SB RAS. The model reaction of cumene oxidation with azobisisobutironitrile as initiator has been carried out at 60 oC. The heat evaluation (W) was proportional to the chain continuation (reaction II) or break of peroxyradicals at the inhibitor-antioxidant (reaction VII):

k1 · · RH + RO 2  ROOH + R (I) k2 · · InH + RO 2  ROOH + In (II)

[AOX ]0 The concentration of the antioxidants can be calculated by the formula: τ = fn , wi where the initial concentration of antioxidants [AOX]0 (mol/l) is related to induction period , and the rate of initiation wi (l/mol∙s), whereas f is a stoichiometric coefficient of inhibition, and n is a number of functional groups. Results. Experimental curves for the initiated cumene oxidation have demonstrated various AOX activities for different objects of investigation. The induction period  was recorded in presence of active antioxidants, for which the AOX concentration and the rate constant of peroxyradical-antioxidant interaction (k2) have been calculated. Generally, the lipid fractions contain the homologues of tocopherols, which act as strong antioxidants. The essential oils and many CO2-extracts contain weak AOX, for which only the decrease in the rate of oxidation wox is determinable. Thus, the proposed microcalorimetric method is a universal procedure for study of AOX- activity of natural lipid fractions of any nature. This accurate and simple-to-use method allows to determine the content of antioxidants and their ability to accept the peroxyradicals.

140 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Oxidation Usnic Acid by Organic Peracids

Dmitry N. Sokolov, Olga A. Luzina, Marina P. Polovinka, Nariman F. Salakhutdinov

N.N. Vorozhtsov Novosibirsk Institute of Organic chemistry SB RAS 630090, Lavrentiev av., 9, Novosibirsk, Russia E-mail: [email protected]

Usnic acid 1 is a secondary metabolite found in several lichen species. It has a wide range of biological activities [1]. Previously described products of the oxidation cleavage usnic acid in solutions of sodium and potassium hydroxide under oxygen stream and the yields were very low [2]. The ring C was oxidized under those conditions. We carried out reactions usnic acid 1 and its pyrazole derivative 4 with organic peracids (peracetic, m-chloroperbenzoic, monoperphthalic). The reactions were carried out with 3 equiv of peracid by mixing the reactants in chloroform at room temperature. The ring A was oxidized in both substrates under these conditions. The yields of compounds 2, 3 and 5 after chromatography were 41%, 27% and 43% respectively. 7,9-Diacetate usnic acid didn’t oxidized by organic peracids.

The structure of compounds 2, 3 and 5 was determined on the basis of their 1H, 13C NMR spectra and X-ray analysis (only 2).

1. Ingolfsdottir, K. Phytochemistry 2002, 61, 729-736. 2. Takani, M., Takahashi, K. Chem. Pharm. Bull 1980. 28, 177-180.

141 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Chemical Contaminations of Camhorosma monspeliacum

A.K. Umbetova, N.A. Sultanova, G.Sh. Burasheva, Zh.A. Abilov

Al-Farabi Kazakh National University, 050012, Karasay batyr 95A, Almaty, Kazakhstan Fax: + 7272 923731, e-mail: [email protected]

On the saline and arid soils of Central Asia, widely distributed some halophytes which belong to family Chenopodiaceae. The genus Camphorosma (Chenopodiaceae) belong to euhalophytes which growing on salt soils along of rivers and lakes. In the flora of Kazakhstan found 3 species of genus Camphorosma: C. monspeliacum, C. Lessingii, C. songoricum which use in folk medicine as stimulating, diuretic and diaphoretic agents. In this work phytochemical and quantitative analyses of biological active complexes of aerial part of C. monspeliacum were done. The amino-, fatty acids and mineral contaminations were determined by using gas-liquid chromatography and atomic absorption spectroscopy techniques accordantly. The influence temperature, duration of extraction process, the ratio of raw materials, solvents and chromatographic parameters were studied. A scheme of separation and isolation of biologically active components were elaborated. In results were isolated 10 compounds such as terpenoids, chromones, flavonoids and phenolic acids. The structure of individual compounds were determinate by chemical (hydrolyses, destruction) and spectroscopy (IR, UV, NMR 13C-, NMR 1H-, 2D-COSY 45?, HNQC, HMBC, mass spectrometry) methods. The different extracts which obtained of C. monspeliacum shown antibacterial and antifungal activities. Above mention results show that the euhalophyte of genus Camphorosma is a new source of biologically active compounds.

142 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation The Separation of Biological Active Complexe from Genus Tamarix by Using Nanostructure Sorbents

a N.A. Sultanova, a G.E. Zhusupova, a Zh.A. Abilov, b Zh.A. Mansurov, b M.A. Beisenbaev

a Al-Farabi Kazakh National University, 050012, Karasay batyr 95A, Almaty, Kazakhstan Fax + 7272 923731, e-mail: [email protected] b RSE Institute of combustion problems, 050012, Bogenbay batyr 172, Almaty, Kazakhstan Fax + 7272 925811

The carbon sorbents widely use successfully in difference industry and medicine for purification some impurity. It is known that in Europe carbon sorbents obtained from seeds of olive and grapes and nutshell. The object of investigation is Tamarix hispida (Tamaricaceae) which growing on salt soils. Early chemical contaminations of Tamarix hispida were investigated. It is known that genus Tamarix is rich source of biological active compounds and most of these are high polar for example sulphates and glycoside forms of polyphenols which poses difference good antibacterial, antioxidant, fungal activities. So for isolation of mention compounds need some effective original sorbents. In this work for separation biological active compounds from aerial part of Tamarix hispida used nanosorbents with developed structure and sorption activity which obtained on the basis apricots and rise shell by the carbonation. The scheme of obtain nanosorbents with using modern technology were elaborated. In results obtained some fractions which contained amino acids, carbohydrates, polyphenols and glycosides of phenolic compounds. The obtain results shown that nanosorbents on the basis industry waste of apricots and rise shell likes polyamide and can use for isolation some individual biological active components.

143 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation The Basidiomycete Lentinula edodes Lectins Interaction with the Selected Organoselenium Diphenyl Substances

Alexei N. Pankratov1, Olga M. Tsivileva2, Boris I. Drevko3, Valentina E. Nikitina2

1Division of Analytical Chemistry and Chemical Ecology, Institute of Chemistry, N. G. Chernyshevskii Saratov State University, 83 Astrakhanskaya Street, Saratov 410012, Russia 2Laboratory of Microbiology, Institute of Biochemistry and Physiology of Plants and Microorganisms, RAS, 13 Entuziastov Ave., Saratov 410049, Russia 3Saratov Military Institute of Biological and Chemical Safety, Saratov, Russia Fax: ++7 8452 970 383 E-mail: [email protected]

Literature data point out the direct participation of microelement selenium in biochemical and physiological processes. However, there is a need for detailed information on the selenocompounds in plants of economic importance, and the use of more sensitive techniques, including those accompanied by theoretical computer-assisted studies, should fill this gap. Among the new pharmaceutical products, the carbohydrate-binding proteins, lectins, isolated from higher basidiomycetes have received the most extensive attention of researches. In relation to the aforesaid, the task of studying the role of spatial and electronic structure, hydrophobic properties and concentration of organoselenium compounds on their interaction with fungal metabolites – extracellular lectins of Lentinula edodes (shiitake mushroom) has been considered. By the hybrid method of density functional theory at the B3LYP/6-31G(d,p) theory level, spatial and electronic structure of the 1,5-diphenyl-3-selenopentanedione-1,5 (preparation DAPS-25), 1,5-di(4-methoxyphenyl)-3-selenopentanedione-1,5 and 1,5-di(4-ethoxyphenyl)- 3-selenopentanedione-1,5 molecules has been studied. The above molecules have been stated to be substantially similar to each other by their electronic and spatial characteristics. By means of the QSAR properties evaluation by the atomic-bonding-additive schemes, it has been shown that the molecules of the preparation DAPS-25, its dimethoxy- and diethoxy- substituted are close to each other by the hydrophilic-lipophilic balance, whereas di-n- octyloxy derivative DAPS-25 is explicitly hydrophobic. The hemagglutinating activity of lectins in the presence of the preparation DAPS-25 and its alkyloxy-substituted increases, therewith the most effective addition is 1,5-di(4- ethoxyphenyl)-3-selenopentanedione-1,5. Apparently, the greater effectiveness of the said substance compared to DAPS -25 is caused by the formation of hydrogen bonds with a participation of unshared electron pairs of oxygen atoms from the ethoxy groups and mobile hydrogen atoms from the OH groups of glycoconjugates on erythrocytes surface. The positive effect of 1,5-di(4-n-octyloxyphenyl)-3-selenopentanedione-1,5 is not so prominent, since the enlarged alkyl chain shields the aromatic fragments of organoselenium molecule participating in the binding with lectin.

144 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation A Preparative Method for Isolation of Fucoxanthin from the East Sea Brown Algae Eisenia bicyclis by Centrifugal Partition Chromatography

Sang Min Kim, Ya Fang Shang, Byung-Hun Um

Natural Products Research Center, KIST Gangneung Institute, Gangneung 210-340, Korea Fax: 82-33-650-7299 E-mail: [email protected]:

A centrifugal partition chromatography (CPC) method was successfully established for isolation of fucoxanthin, a major carotenoid of brown algae, from an Eisenia bicyclis extract using a two-phase solvent system of n-hexane:ethyl acetate:ethanol:water (5:5:7:3, v/v/v/v). A preparative CPC yielded 20 mg of fucoxanthin in a two step separation from 516 mg of a fraction containing 4.59% fucoxanthin (Fuco-fraction). The purity of the isolated fucoxanthin was over 98%, as determined by high performance liquid chromatography (HPLC). The structure of fucoxanthin was confirmed with APCI/MS and NMR.

145 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Preparative Separation of Chlorogenic Acid by Centrifugal Partition Chromatography from Highbush Blueberry (Vaccinium corymbosum L.)

Sang Min Kim, Ya Fang Shang, Byung-Hun Um

Natural Products Research Center, KIST Gangneung Institute, Gangneung 210-340, Korea Fax: 82-33-650-7299 E-mail: [email protected]:

Blueberries (genus Vaccinium) have gained worldwide focus because of the high anthocyanin content of their fruits. In contrast, the leaves of blueberry have not attracted any attention, even though they contain large quantities of chlorogenic acid, a strong antioxidant compound. The aim of this investigation was the quantification and preparative isolation of chlorogenic acid from blueberry leaves using a new separation scheme, Centrifugal Partition Chromatography (CPC). A water fraction containing a high concentration of chlorogenic acid was obtained by defatting a crude methanol extract from blueberry leaves. Centrifugal partition chromatography (CPC) was applied to isolate chlorogenic acid from this water fraction using a two-phase solvent system of ethyl acetate-ethanol-water at a volume ratio 4:1:5 (v/v/v). The structure of chlorogenic acid in the CPC fraction was confirmed with HPLC, UV, ESI/MS and NMR spectra. The HPLC chromatogram showed hat t the fractions collected by CPC contained chlorogenic acid with 96% purity based on peak area percent. The total amount of chlorogenic acid isolated from 0.5 g of a water fraction was 52.9 mg, corresponding to 10.6% of the water fraction. Chlorogenic acid was successfully isolated from blueberry leaves by the CPC method in a one step procedure, indicating a further potential use for blueberry leaves.

146 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Anthocyanins from Azalea (Rhododendron mucronulatum) Flowers

Je Seung Jeon, Byung-Hun Um

Natural Products Research Center, KIST Gangneung Institute, Gangneung 210-340, Korea Fax: 82-33-650-7299 E-mail: [email protected]:

The azalea (Ericaceae family) flower has blossomed before shooting its leaves, in early spring. Especially in Korea, the azalea flower has been made into a wine called “Doo gyeun joo wine”, and it was used as folk medicine for tonic, diuretic, and expectorant, as well as for its depressant effect. A new anthocyanin, petunidin-3-O-α-L-arabinosyl-5-O-β-D-glucoside(1), was isolated from the flowers of Korean Aazalea (Rhododendron mucronulatum) together with six known anthocynins and three antioxidant flavonoids, malvidin-3-O-α-L-arabinosyl-5-O-β-D- glucoside(2), cyanidin-3-O-α-L-arabinosyl-5-O-β-D-glucoside(3), cyanidin-3-O-α-L- arabinoside(4), malvidin-3-O-α-L-arabinoside(5), delphinidin-3-O-α-L-arabinoside (6), malvidin-5-O-β-D-glucoside(7), myricetin(8), quercetin(9), kaempferol(10). Their structures were identified by spectroscopic method, including 1D- and 2D-NMR. Our findings can be used as a foundation for further chemotaxonomic studies on Ericaceae family flowers.

147 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation The rapid identification of anthocyanins from blueberry, Vaccinium corymbosum by LC–NMR/MS

Suk Woo Kang, Sang Min Kim, Byung-Hun Um

Natural Products Research Center, KIST Gangneung Institute, Gangneung 210-340, Korea Fax: 82-33-650-7299 E-mail: [email protected]:

The anthocyanins and caffeoylquinic acid are major bioactive compounds in blue berry fruits, and the kinds and contents of anthocyanins are greatly changed in its cultivars and cultivation environment. In common HPLC analysis methods, the standard compounds are absolutely needed for the identification of these anthocyanins. In this study, the hyphenated LC-NMR and LC-MS techniques are applied for the direct identification of the major constituents present in buleberry. The hyphenated techniques of LC-NMR and LC-MS are increasingly used for phytochemical analysis. These techniques combine the separation power of HPLC with the structural information provided by NMR and MS. The major advantages over the traditional off-line techniques are rapidity and efficiency. Reversed-phase chromatography was performed using an acetonitrile-water solvent system on a C18 column. The NMR spectrum yielded six main peaks, whose analysis revealed them to be delphinidin-3-galactoside (1), chlorogenic acid (2), delphinidin-3- arabinoside (3), petunidin-3-galactoside (4), malvidin-3-galactoside (5), and malvidin-3- arabinoside (6). The identities of these constituents were confirmed by performing LC-ESI- MS. This study shows that hyphenated LC-NMR and LC-MS can be used to rapidly (55 min) identify anthocyanins.

148 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation To the Question of Standardization of Anticancer Drug «Leykoefdin» on the Basis of the Herb of Ephedra Equisetina

Yevgeniya S. Ushakova

Al-Farabi Kazakh National University, Karasay batir str., 95A, Almaty, 050012, Kazakhstan, E-mail: [email protected]

Provision of Kazakhstan's population of domestic production of drugs is one of the main priorities of social policy of the Republic of Kazakhstan. The successful solution of this problem is closely linked with the development of natural wealth, the creation of highly effective natural and synthetic drugs, the elaboration the universal analyzed samples, and highly sensitive methods for the determination of active substances and by-products. The aim of this study was to develop methods of standardization of highly effective anticancer drug “Leykoefdin”, developed at the Department of Chemistry of Natural Compounds of Al-Farabi Kazakh National University on the basis of the herb of ephedra equisetina. “Leykoefdin” shows a high antitumour effect on the source, induced drug-resistant variants of tumours and their metastases. The drug is nontoxic, does not oppress hematopoietic, liver, kidney, endocrine, cardiovascular, central and peripheral nervous systems, nonimmunodepressive, no allergic. When intravenously Pliss lymphosarcoma (92% inhibition), sarcoma 180 (89%), Ehrlich solid tumour (84%), liver cancer ACL-1 (70%), Guerin's carcinoma (65%), Walker carcinoma (65%), etc. According to the literature and the results of the bioscreening of the components of “Leykoefdin” held in Kazakh Research Institute of Oncology and Radiology, active drug substances are oxidized forms of flavonols. Synergism of flavonoid compounds on the above strains also show hydrolyzed tannins based on the ellagic and m-digall acids and leykoantotsianidins. For the quantitative determination of active substances of the drug, we have used a spectrophotometric method based on the interaction of drug with 1% alcoholic solution of aluminum chloride, with formation the colored complex. As the comparison solution was used 95% alcohol solution of the drug. We have also studied the influence of pH, temperature and reagent concentrations on the accuracy and reproducibility of the results of the analysis of the chosen method. It was found that the formed complex is stable up to temperature 70oC. Application of the method is acceptable in weakly acidic and neutral solutions, at pH values higher than 1-2 units of pH of working solution (4.95). In determining the concentration dependence was revealed that with decreasing concentration the role of competing reactions decreases, and thus, the quantitative content of flavonoids increases (Table): Concentration, Quantitative content of flavonoids, Concentration, Quantitative content of flavonoids, % % % % 0,25 17,53 0,03125 39,34 0,125 28,87 0,015625 42,91 0,0625 31,90 The advantages of the offered method are the high accuracy of the results, ease of performance, availability, the opportunity to experiment with nonliofilizied drug. The disadvantage is a significant number of used reagents.

149 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Investigation of Standardization of Rosa corymbifera Phytopreparation

N.A. Vereshchagin

Al-Farabi Kazakh National University, Almaty E-mail: [email protected]

Rosa corymbifera is well-known medicinal plant part of pharmacopoeias in many countries. The rosehips is generally acknowledged medication in Chine, Tibet, India, Mongolia and Siberia. Rosehip is not only a healing agent, it also potentiates the components in complex mixtures. Basic active ingredient are hardening agents of hydrolysable and condensed type – flavonoids, polysaccharides and ascorbic acid. Rosehip pulp contain riboflavin, beat-carotene, phylloquinone and bioflavonoids and the seeds contain tocopherol, carotene and fatty oil, 14.1% of pectic substance, 1.58% of citric acid. Such substances as meletin, kaempferol, isoquercitrin, tilirozid, epigallocatechin, gallocatechin, epigallocatechingallate, epicatechingallate were found in rosa corymbifera. Dried rosehips and cumulative phytopreparation of rosa corymbifera (manufactured by

KhimFarm JSC) have been investigated. Based on experiment data for chemical composition of phytopreparation of rosa corymbifera and literature data on immunopotentiating and vitamin activity of secondary metabolite of the plants, we proposed to standardize medication based on quantitative and qualitative method of verification of active ingredients – hardening agents of condensed type, polysaccharides and ascorbic acid; with regard to additive composition – oxidated forms of flavonoids. We investigated both official pharmacopoeial methods for authenticity control of individual bioactive substances and data from pharmacopoeial articles and TAND for cumulative phytopreparations of phytogenic origin registered in the Republic of Kazakhstan. We investigated temperature and concentration limits of reactions for selected methods of authenticity control of phytopreparations; studied impact of pH to the character and reaction direction. We have selected reactions with ammonium iron alums (hardening agents), ferric chloride solution, lead acetate solution, vanillin solution in concentrated hydrogen chloride, concentrated ammonia (flavonoids), ferricyanide solution (ascorbic acid), α-naphthol solution

150 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation in ethylene alcohol, resorcine solution in ethylene alcohol (polysaccharides). The data is summarized here below:

Concentration Temperature Reagent Impact of pH to reaction limit, % limit, 0C Ammonia 0,0625 0-100 No substantial alteration Ferrous chloride (III) 0,03125 0-70 No substantial alteration Iron-ammonium alum 0,03125 0-70 No substantial alteration Uprating of pH stimulates Lead acetate 0,125 0-100 illustrative reaction process Vanillin 0,125 0-100 Uprating of pH increases reaction Ferricyanide 0,03125 20-80 No substantial alteration Reaction is performed adding α-naphthol 0,125 0-100 concentrated sulfuric acid Reaction is performed adding Resorcine 0,0625 0-100 concentrated sulfuric acid

High temperature limit of ferric iron with chloride is stipulated by rapid oxidation of phenol compositions under temperature + 700C. Temperature limit for reaction with ammonium iron alums is stipulated by imbalance of chelate complex which is developed in high temperature environment. Low speed of oxidation-reduction reaction with ferricyanide is stipulating it's low temperature limit, whereas ascorbic acid oxidation caused by increased temperature provides high temperature limit for reaction. Definition of concentration limits of reactions was performed based on contrast of transit of coloration of working solutions. Based on experiment data we may state that most sensitive, precise and reproductible method of authenticity control of phytopreparations are reactions with ammonium iron alums (hardening agents), concentrated ammonia (flavonoids), ferricyanide (vitamin C), α-naphthol (polysaccharides).

151 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Perspectives of Production of Supplement with Antitumor Effect, Based on Polysaccharides from Brown Algae

Olesya S. Vishchuk1, Svetlana P. Ermakova1, Thinh D. Pham2, Bui Minh Ly2, Tatyana N. Zvyagintseva1

1 Pacific Institute of Bioorganic Chemistry, FEB RAS, 159, 100-letya Vladivostoky, 690022, Vladivostok, Russia 2 Nha Trang Institute of technology research and application, Vietnamese Academy of Science and Technology, 02 Hungvuong st., Nha Trang City, Vietnam Fax: 8(4232)-31-07-05 E-mail: [email protected]

Cancer is known to be one of the most diseases that badly threaten human’s life. Unfortunately drugs, which are used for cancer therapy, are toxic and destroy not only cancer cells but normal cells and tissues. That’s why search and production of new effective nontoxic compounds from natural sources is actual problem. At last decade marine brown algae attract much attention because they represent a rich and easily regenerated source of polysaccharides with interesting structures and biological activities: alginic acids, laminarans and fucoidans [1]. The sulfated polysaccharides (fucoidans), which mainly build up of α-L-fucose residues and sulfate groups, have a wide variety of biological activities, including anticoagulant, antivirus, immunomodulating and antitumor activities [2]. But because the structures of fucoidans are complex and heterogeneous and as many studies of biological activity were carried out using relatively crude polysaccharide preparations, it is presently not easy to determine the overall relationship between activity and structure of polysaccharides. The aim of present work is investigation of antitumor activity of fucoidans with different structure, isolated from brown algae of the Sea of Japan, of the Sea of Okhotsk and of the South China Sea. Fucoidans were isolated from different types of brown algae (Laminaria cichorioides, L. japonica, Fucus evanescens, Undaria pinnatifida, Sargassum swartzii, S. oligocystum, S. denticarpum, S. mcClurei, S. polycystum) as described earlier [3]. The structural characteristics of polysaccharides were elucidated by chemical and physic- chemical methods, including 13C-NMR and mass-spectrometry. It was shown that fucoidans from different types of brown algae were differed in degree of sulfation, monosaccharide composition and glycosidic bonds of main polysaccharides’ chain. Investigation of antitumor activity of fucoidans showed that they were non-toxic on JB6 mouse epidermal cells in concentrations of up to 200 ug/ml and inhibited formation and progression of colonies of HT-29 and DLD-1 colon cancer cells in dose-dependent manner. Moreover, completely sulfated fucoidan from L. cichorioides, consisted of 1,3-linked α-L- fucose residues, possessed the highest antitumor activity, compared with fucoidans from other types of algae. Taken together, these results indicate that fucoidans from brown algae exert potent antitumor activity and they are certain to be perspective base for production of supplement with antitumor effect.

1 Zvyagintseva T.; Shevchenko N.; Nazarenko E. et.al. J. Exp. Marine Biology and Ecology 2003, 294, 1-13 2 Li B.; Lu F., Wei X.; Zhao R. Molecules 2008, 13, 1671-95. 3 Shevchenko N.; Kysaikin M.; Urvantseva A. and et. al. Patent RU 2240816 2004, 33, 444-445.

152 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Biocompozites on the Base of Natural Sorbents, Synthetic and Natural Polymers

I.A. Vorsina, T.F. Grigorieva, A.P. Barinova, N.Z. Lyakhov

Institute of Solid State Chemistry and Mechanochemistry SB RAS, Novosibirsk, Russia E-mail: [email protected]

Mechanochemical approach to the performance of reactions in organic chemistry not only ecological and economical advantages over traditional technologies have but also allow obtaining novel substances with required properties that cannot be synthesized in the liquid- phase version of the process. We obtained dispersed mechanocomposites on the base layered silicates and aerosil containing biologically active organic acids with different nature. For example: succinic, citric, tartaric acids, α-glycine. In case of layered silicates mechanochemical reaction of neutralization proceeds. The anions of acids are chemically bound with the metal ions of silicates. The acid anions chemically bound to the carrier surface get detached from the carrier rather easily in a weakly acidic medium (pH ~ 1.0) and pass into the solution. Preparation of medical products using the mechanochemical method will allow one to increase the concentration of an active component in the product substantially without any increase in the negative effect on the human organism because bionanocomposites. During the joint mechanical activation of aerosil with organic acids the formation of active acids centre on the surface of silicate takes place. The simultaneous break of H-bonds in association of organic acids lead to formation of mechanocomposites with H-bonds between aerosil and monomeric acids molecules. It is established that during mechanochemical interaction of organic acids with synthetic medicinal polymer poly-N-vinylpyrrolidon mechanocomposites of polymer with monomeric molecules acids were formed by means of H-bonds. Soluble natural biopolymers, arabinogalactan (AG) exhibits a vide range of immunobiological activity. One of the most important properties of AG – the ability to penetrate through membrane barriers of cells. It follows from the analyses of the experimental data that only polybasic and hydroxy acids interact with AG by forming H-bonds between the protons of carboxylic groups and oxygen atoms of unsubstituted vicinal hydroxyl groups of the side chains of AG. A number of formulas of curative powders and the technologies of their manufacture were developt (Authors` Certificate No. 1754102 of 29.06.89). experimental clinical test were carried out at the Moscow Beauty Institute; a permission from the State Committee of Sanitary and Epidemiological Inspection for their introduction into industrial production was obtained. The data obtained may be used in developing new pharmaceutical dosage forms, nutrition additiver and preparations of therapeutic and prophylactic cosmetics.

153 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Molecular Characteristics of Water-Soluble Sulphated Polysaccharides from Enteromorpha prolifera and Their Anticancer and Immunomodulatory Activities

Myeong Lae Cho and Sang Guan You

Department of Marine Food Science and Technology, Gangneung-Wonju National University, 120 Gangneung Daehangno, Gangneung, Gangwon, 210-702, Korea Fax: 82-33-640-2850 e-mail: E-mail: [email protected]

Water-soluble sulphated polysaccharides were isolated from Enteromorpha prolifera and fractionated using an ion-exchange chromatography to investigate their chemical and molecular characteristics as well as their anticancer and immunomodulatory activities. The crude and fractionated polysaccharides (F1, F2 and F3) mostly consisted of carbohydrates (50.0-62.5%) and sulphates (14.5-18.8%) with different levels of various monosaccharides including rhamnose (57.1-87.6%), glucose (3.6-39.1%) and xylose (2.4-8.8%). The protein contents were varied from 1.0 to 13.9% and after the fractionation, most proteins were included in F2 fraction (11.3%). F1, F2 and F3 contained two or three subfractions with the 3 weight average molecular weights (Mw) ranging from 37.4 to 1,21810 g/mol. The relatively low in vitro anticancer activity of the fractions (1.2-26.1%) suggested that they had no significant direct cytotoxicity to AGS and DLD-1 cells. On the other hand, a proteo- heteroglycan (F2 fraction) exhibited a strong immunomodulatory activity, comparable to the positive control, LPS.

154 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Coagulation Ability of Humic Acids from Mechanochemically Activated Peat

Alexander A. Ivanov1, Natalya V. Yudina1, Oleg I. Lomovsky2

1 Institute of Petroleum Chemistry SB RAS, Akademicheskiy pr. 4, Tomsk, 634021, Russia E-mail: [email protected] 2 Institute of Solid State Chemistry and Mechanochemistry SB RAS, ul. Kutateladze 18, Novosibirsk, 630128, Russia

The rheological properties of blood are known to depend substantially on the erythrocyte reversible aggregation. Aggregation parameters may be employed as universal indicators for determining the coagulation and stabilization ability of natural preparations. Among many factors providing blood with coagulability, such coagulants as vitamin K and naphthoquinone derivatives play an important role. The goal of this work is to study erythrocyte reversible coagulation under the effect of humic acids (HAs) isolated rom f peat subjected to a mechanochemical treatment. High-moor peat treatment was carried out in a flow-type vibrating-centrifugal mill in the presence of a dry alkali or a cellulolytic ferment. Erythrocyte reversible aggregation was studied with an instrument using the vibrational photometric method of measuring the optical density in microscopic volumes of blood1. Erythrocyte reversible aggregation was investigated in alkaline solutions at HAs concentrations of 0.001 and 0.0001 wt %. Results of researches have shown, that peat mechanical activation increases the yield and solubility of humic preparations. The comparison between results on erythrocyte reversible aggregation testifies that the highest effect is achieved with HAs prepared through the peat mechanical activation, either in the absence of additives or in the presence of cellulolytic enzyme. A rise in the concentration of HAs increases their coagulation ability. The special methods of HAs modification during controlled mechanochemical oxidation and enhancing of oxygen-containing fragments, including chelate groups, are developed. The results of the structural analysis enables us to conclude that mechanical treatment enlarges the amount of hydrophilic oxygen-containing fragments and slightly decreases the number of paramagnetic centers in HAs molecules.

1. Tukhvatulin, R.T.; Novikova, L.K.; Anosova, N.V.,Uspekhi Fiziolog. Nauk, 1994, vol. 25, № 4, 95.

155 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Bismuth Compounds for Medicine Based on Natural Products

Yuri M. Yukhin, Olga A. Logutenko, Evgeniya V. Timakova, Ekaterina S. Naydenko, Lubov’ I. Afonina, Tatiana A. Udalova

Institute of Solid State Chemistry and Mechanochemistry SB RAS, Kutateladze,18,630128, Novosibirsk, Russia Fax: (383)3322847 E-mail: [email protected]

Bismuth compounds with gallic, tartaric, benzoic and citric acids are widely used in medicine. All these acids are produced from natural products. The main requirements for compounds used in pharmacy are a low content of impurities and high therapeutic activity. Synthesis of bismuth (III) compounds is usually carried out by precipitation from bismuth nitrate solutions which are prepared by dissolution of bismuth metal (usually Bi 1 grade metal, containing not less 98.0 % Bi) in nitric acid solutions of 7-9 mol/l concentration. However, this method does not always allow production of a high purity compound because major metal impurities precipitate together with bismuth. That is why investigations of the choice of optimum conditions for bismuth purification from metal impurities were carried out and the compositions of the bismuth (III) salts so produced were determined. In this work a comparison between the two ways of synthesis of bismuth compounds – by precipitation from nitrate solutions and based on the interaction between solid bismuth . oxohydroxobismuth (III) nitrate trihydrate [Bi6O5(OH)3](NO3)5 3H2O (OHNB) and acid solutions – was made, and the product compositions were determined. Bismuth (III) subgallate and citrate, irrespective of the way of synthesis, were shown to have compositions

C6H2(OH)3COOBiO∙3H2O and BiC6H5O7, respectively. The interaction of OHNB with the solutions of tartaric acid results in the formation of the bismuth (III) tartratahydrotartrate trihydrate [Bi(C4H4O6)(C4H5O6)]·3H2O. Bismuth precipitation from nitrate or perchlorate solutions with sodium benzoate, dependent of the synthesis conditions, can result in the formation of neither bismuth oxide-benzoate BiOC7H5O2 or bismuth tribenzoate Bi(C7H5O2)3. To produce high purity bismuth (III) compounds from impure products, the synthesis based on the interaction between OHNB and the acid solutions would be appropriate for use.

156 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Preparation of Monoclonal Antibodies Against the HBsAg from Transgenic B. mori Larvae

Elvira G. Yusupova

Institute of Chemistry of Plant Substances, Mirzo-Ulugbek Str. 77, 100170 Tashkent, Republic of Uzbekistan Tel: (0998)2622099, Fax: (0998) 1206475, E-mail: [email protected]

Hepatitis B virus (HBV) infection is a global health problem. Two billion people have been infected worldwide; 360 million suffer from chronic HBV infection, and over 520,000 die each year (50,000 from acute hepatitis B and 470,000 from cirrhosis or liver cancer) (Kane, M. 1995. Global programme for control of hepatitis B infection. Vaccine 13 (Suppl. 1):S47-S49). HBV surface antigen (HBsAg) is the established serological marker used routinely for the diagnosis of acute or chronic HBV infection, the screening of blood or organ donors, and the surveillance of persons at risk of acquiring or transmitting HBV. The aim of this investigation is obtaining of hybridomas secreting monoclonal antibodies reactive to HBsAg. For this purpose mice were immunized with partly purified recombinant HBsAg obtaining by using of baculovirus technology from transgenic B. mori larvae. The fusion of spleen cells of immunized mice with the mouse myeloma cell line X Ag 8.653 provided according standard method. Standard sandwich ELISA methods were used for screening and measurement of anti-HBs. Positive hybridomas were subsequently cloned by limiting dilution and specific clones selected. A total of 16 growing hybridomas secreting antibodies reactive to HBsAg, were established.

157 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Retina Protection Properties of 4-Methyl-2,6-diisobornylphenol

Anna A. Zhdankina1, Galina A. Kon1, Mark B. Plotnikov2*, Sergey V. Logvinov1*, Alexander V. Kuchin3*, Irena Yu. Chukicheva3*

1 Department of Histology, Embryology and Cytology, Siberian State Medical University, Moscowsky tract, 2, 634050, Tomsk, Russia 2 Institute of Pharmacology, Siberian Department, Russian Academy of Medical Sciences, Lenin street, 3, 634028, Tomsk, Russia 3 Institute of Chemistry, Komi Scientific Center, Ural Department, Russian Academy of Sciences, Pervomaiskaia street, 48, 167982, Syktyvkar, Russia Fax: 8(3822)41-42-35 E-mail: [email protected]

The derivative of o-isobornylphenol 4-methyl-2,6-diisobornylphenol has high antioxidant, hemorheological, antiplatelet and antithrombogenic properties [1]. The aim of this research is to study the retina protection properties of 4-methyl-2,6- diisobornylphenol in OXYS rats with chorioretinal degeneration. Retinas of 40 male OXYS and Wistar rats at the age of 6 months were studied. The rats in experimental group (n=15) have received 4-methyl-2,6-diisobornylphenol p.o. in the dose of 100 mg/kg daily for 1,5 months. Vascular disorders (decrease of open functioning vessels number of choroid and retinal neurodegeneration) are prevalented in OXYS rats. Course p.o. administration of 4-methyl- 2,6-diisobornylphenol induced significant two-fold increased of open vessels area, five-fold area and high preservation of the retinal neurons. Distinct retina protection effect of 4-methyl-2,6-diisobornylphenol opens perspectives for use of this substance in preventation and treatment of free radical pathologies in ophthalmology.

1 Plotnikov M.B.; Ivanov, I.S; Smoljakova V.I.; et al. Chemistry and Medicine: Abstracts of Proceedings. VI - Russia Scientific Symposium 2007, 80–81.

158 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation New Flavonol Glycoside from Limonium gmelinii

Zhanar A. Kozhamkulova1, Mohamed M. Radwan2, Galiya E. Zhusupova1, Zharylkasin A. Abilov1 and Samir A. Ross2,3

1 al-Farabi Kazakh National University, Karasai Batyra 95a, 050012, Almaty, Kazakhstan 2 National Center for Natural Products Research, School of Pharmacy, University of Mississippi, 38677, USA 3 Department of Pharmacognosy, School of Pharmacy, the University of Mississippi, 38677, USA E-mail: [email protected]

Gmelinoside I (1), a new flavonol glycoside, was isolated and chemically characterized as 3,5,7,3`,4`,5`-hexahydroxy-3-O-α-D-galactopyranosylflavone from the aerial part of Limonium gmelinii (Plumbaginaceae) by UV, IR, ID, 2D NMR and HRESIMS experiments. 25 Compound 1 was isolated as an optically active yellow amorphous powder. [α]D -16 (c 0.05 MeOH); Rf: 0.58 (Si, EtOAc-HOAc: H2O, 5:1:1). Its HRESIMS showed an [M+H] ion peak at m/z 481.09822 corresponding to the molecular formula C21H20O13. UV (MeOH): λmax 265,

365 nm, UV (NaOAc): λmax 271, 307, 346, UV (AlCl3) 279, 310, 430 nm. The UV spectrum of 1 in MeOH displayed two peaks at λmax 265 nm (Band I) and 365 nm (Band II) characteristic for a flavonol skeleton [1]. The bathochromic shift of Band I with the separate addition of NaOAc (6.0 nm) and AlCl3 (14.0 nm) to the methanolic solution of 1 indicated the 1 hydroxylation at C-7 and C-5. H NMR (DMSO-d6, 400 MHz): δH 3.33 (2H, m, H-6"), 3.38 (1H, m, H-3"), 3.45 (1H, m, H-5"), 3.61 (1H, m, H-2"), 3.64 (1H, m, H-4"), 5.33 (IH, d, 0.8 Hz), 6.19 (1H, s, H-6), 6.37 (1H, s, H-8), 7.19 (1H, s, H-6`), 7.21 (1H, s, H-2`), 12.63 (1H, s, 1 OH-5). The H NMR spectrum of 1 showed four aromatic singlets at δH 6.19, 6.37, 7.19, 7.21 nm corresponding to H-6, H-8, H-6', and H-2', respectively. It also showed an anomeric proton at δH 5.33 (d, J = 0.8 Hz, H-1") correlated to δC 102.1 (C-1") in the HMQC spectrum indicated a monoglycosylation which was confirmed by the presence of four oxymethine (δC 68.0-75.9) and one oxymethylene (δC 60.0) carbons. The site of glycosylation was determined 13 by the HMBC correlation of H-1" (δH 5.33) and C-3 (δC 133.7). C NMR ((DMSO-d6, 100 MHz): δH 177.4 (C-4), 164.2 (C-7), 161.2 (C-5), 156.2 (C-9), 145.4 (C-3`, C-5`), 136.7 (C- 4`), 133.7 (C-3), 119.9 (C-1`), 108.5 (C-2`,C-6`), 103.8 (C-10), 102.1 (C-1"), 98.7 (C-6), 93.3 (C-8), 75.9 (C-3"), 73.3 (C-5"), 71.2 (C-2"), 68.0 (C-4"), 60.0 (C-6"). The 13C NMR spectroscopic data of the sugar were quite similar to those reported for D-galactosyl moiety [2-3], but the small coupling constant of the anomeric proton (J = 0.8 Hz) indicates its α configuration, not β configuration [3-4]. This sugar was identified as α-D-galactopyranose and the structure of 1 was elucidated as 3,5,7,3',4',5'-hexahydroxy-3-O-α-D- galactopyranosylflavone or gmelinoside I.

1 Mabry J.; Markham K.; Thomas M. The systematic identification of flavonoids. Springer-Verlage, New- York, Heidelberg, Berlin. 1970. 2 Agrawal P. Carbon-13NMR of flavonoids, Studies in Organic Chemistry. New York. 1989. 3 Gao Z.; Ali Z.; Khan I. Phytochem. 2008, 69, 2856-2861. 4 Kagan J. Phytochem. 1967, 6, 317-318.

159 2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology” March 15-18, 2010, Novosibirsk, Russia Poster Presentation Isolation and Identification of 26-Hydroxyecdysteroids

Larisa Zibareva1, Rene Lafont2, Laurence Dinan2

1 Laboratory of phytochemistry, Tomsk State University, 634050 Tomsk, pr. Lenina, 36, Russia, E-mail: [email protected] 2 Laboratoire de Biochimie structurale et fonctionnelle des Prote?ines, CNRS FRE 2852, Universite?Pierre et Marie Curie, Case 29, 7 Quai Saint Bernard, 75252 Paris, Cedex 05, France

Phytoecdysteroids are analogues of arthropod steroid hormones occurring in plants, where they are believed to contribute to the defense of the plant against invertebrate predation. Analysis of the seeds of a large number of species for the presence of ecdysteroids has revealed more 100 species of higher plants contain these compounds. Silene (Caryophyllaceae) is one of the richest ecdysteroid-containing genera so far detected in the plant world. Commonly, the ecdysteroid-positive Silene species possess a rich ecdysteroid composition: high levels of 20-hydroxyecdysone, accompanied by 2-deoxy-20- hydroxyecdysone, 2-deoxyecdysone, polypodine B and integristerone A. Besides, this genus is characterized by the presence of many new compounds which have been detected only in these plants – ecdysone-22-sulfate, silenosterone, 2-deoxycdysone 22-D-glucoside and other. At present, some 26-hydroxylated ecdysteroids are known, but most of them were found in Silene species. The phytoecdysteroid profiles of extracts of aerial parts of flowering plants of 3 ecdysteroid-containing species in the genus Silene: S. psevdotites, S. fridvaldszkyana, S. viridiflora have been examined and identified by HPLC and, n i the case of four new compounds, by mass spectrometry and NMR.

2-deoxy-20,26-dihydroxy- 20,26-dihydroxyecdysone 26-hydroxyintegristerone A ecdysone 3,22-diacetate New 26-hydroxyecdysteroids – 2,22-diacetate 20,26-dihydroxyecdysone, 3,22-diacetate 20,26-dihydroxyecdysone were detected in Silene viridiflora; 2-deoxy-20,26- dihydroxyecdysone in S. psevdotites; 26-hydroxyintegristerone A in S. frivaldszkyana. Silene viridiflora and S. frivaldszkyana are richer sources of ecdysteroids, synthesized many other 26-hydroxylated compounds. Comparison with the 1H and 13C NMR spectra for 20-hydroxyecdysone reveals changes in the A-ring and side-chain, both compatible with the presence of extra hydroxyl groups (26- hydroxyintegristerone A), since the 13C signals for C-1 and C-26 are significantly shifted to higher frequencies (from 38.09 ppm to 76.9 ppm and from 30.10 ppm to 71.3 ppm, respectively). This is supported by the absence of one of the H-1 signals and the higher frequency shift in the remaining H-1 signal (to 3.93 ppm), together with the absence of a 1 methyl signal and the appearance of –CH2OH signal (3.47 ppm) in H NMR.

160 Authors Index

Authors Index

Baryshnikov, Sergey V...... 206 A Baser, K.H.C...... 72 Abdullaev, N.D...... 92, 94 Bayandina, Irina I...... 20, 73 Abdullaeva, D.U...... 156, 162 Beisebekov, M.K...... 116 Abdurazakov, A.Sh...... 5 Beisenbaev, M.A...... 219 Abilov, Zharylkasin A.36, 58, 116, 148, Bektimirov, A.M.-T...... 184 218, 219, 240 Belovodskii, Alexander V...... 75 Adekenov, Sergazy M.1, 95, 117, 166, Beregovtsova, Natalia G...... 206 182, 194, 208 Beshley, I.V...... 145, 147 Afanasyeva, S.V...... 60 Bobaev, I.D...... 184 Afonina, Lubov’ I...... 236 Bobakulov, Kh.M...... 94 Agafoncev, Alexandr M...... 102 Boldyrev, Vladimir V...... 150, 170 Agzamova, M.A...... 62, 107 Boyarskikh, Irina G...... 77 Aisa, Haji Akber...... 31 Boymatov, I.M...... 184 Akhmedjanova, V.I...... 63 Bozorov, Kh.A...... 5 Akhmedkhodjaeva, Kh.S...... 119 Bravova, I.M...... 89 Aksyonov, Vladimir V...... 9 Budaeva, Vera V...... 22, 79 Alekseev, Alexander A...... 65 Bugaev, Ivan M...... 80 Alekseeva, L.I...... 66 Burasheva, G.Sh...... 31, 218 Aliev, Oleg I...... 176 Bychkov, A.L...... 81 Alikulova, M.U...... 159 Bystrushkin, A.G...... 66 Al-Jaber, Nabila Abduaziz...... 10 Angenot, L...... 63 C Anischenko, Anna M...... 67, 110 Chadin, Ivan F...... 53 Antimonova, Anna N...... 68 Chernenko, Galina F...... 212 Anufriev, Victor Ph...... 39 Chernov, Sergey V...... 166 Ardashov, O.V...... 70 Chernyak, Elena I...... 3, 83, 170 Aripova, S.F...... 119, 143, 159, 160 Chibiryaev, Andrey M...... 33 Arystan, Leila I...... 214 Cho, Myeong Lae...... 233 Asanova, G.K...... 208 Choi, Seon-Kang...... 23 Atazhanova, Gayane A...... 11, 194 Chudinov, A.V...... 89 Avdeev, S.M...... 48 Chukicheva, Irena Yu...... 239 Awaad, Amani S...... 12 Chung, Donghwa ...... 24 Azimova, Shakhnoz S...... 41 Azizov, I.K...... 92 D Davidova, Victoria N...... 54 B Davlatov, N.Sh...... 119 Babkin, V.A...... 13 Dinan, Laurence...... 242 Bacharov, Dmitry S...... 178 Domnina, Nina S...... 176 Baev, D.S...... 46 Domrachev, Dmitry V...... 85, 87, 141 Baisalova, G.Zh...... 72 Drebushchak, T.N...... 150 Bakhvalov, Oleg V...... 28 Drebushchak, V.A...... 150 Balandina, G.N...... 193 Drevko, Boris I...... 220 Balashova, Maria V...... 14 Dunaevsky, Y.E...... 193 Baltina, L.A...... 16, 125 Dyshlovoy, Sergey A...... 25 Barabanova, Anna O...... 18, 54 Barinova, A.P...... 231

161 Authors Index

E K Efremov, Alexander A...... 37, 98 Kabdraissova, Aissulu Zh...... 56 Elmuradov, B.Zh...... 5 Kadutskii, Aliaksei P...... 112, 114 Ermakova, Svetlana P...... 27, 229 Kamoldinov, H.Sh...... 91 Ermatova, A.B...... 60, 89 Kanarskiy, Albert V...... 129 Eshbakova, K.A...... 90, 91, 92, 94 Kang, Suk-Woo ...... 6, 225 Eshmagambetova, A.B...... 208 Kanitar, Kanitar ...... 56 Karpova, Elena V...... 87, 105, 187 F Kashevarov, Nikolay I...... 174 Fedorov, Sergey N...... 25 Kasymbekov, A.A...... 116 Folmer, Vladimir V...... 95 Kazantsev, Alexander V...... 166 Fomenko, Vladislav V...... 28, 97 Khabarov, Ilya A...... 95, 117 Khalilova, E.Kh...... 119 G Khanina, Minisa A...... 141 Gadetskaya, Anastassiya...... 148 Kharitonov, Yurii V...... 120, 154 Galyautdinov, Ilgiz V...... 52 Khidyrova, N.K...... 143, 205, 215 Ganbaatar, J...... 29 Khokhrina, Ekaterina A...... 104 Gapparov, A.M...... 159 Khudobko, M.V...... 16 Gatilov, Yurii V...... 127, 168, 194 Khujaev, V.U...... 159 Genaev, Alexander M...... 105 Khushbaktova, Zainab A...... 109, 205 Golubev, Sergey V...... 98 Kim, Sang Min...... 6, 222, 223, 225 Golyazimova, O.V...... 100, 172 Kim, Yong Hwan...... 54 Gontova, Anna A...... 37 Klochkov, S.G...... 32, 60, 89 Gorshkov, Nikolay B...... 102 Kochetov, A.V...... 3 Grazhdannikov, Alexander E...... 104 Kodirova, D.R...... 122 Grigor’ev, I.A...... 3 Kogay, Tamara I...... 123 Grigorieva, T.F...... 231 Kolchanov, N.A...... 3 Gruzdev, I.V...... 66 Komarova, Nina I...... 163, 185 Kon, Galina A...... 239 H Kondratenko, R.M...... 125 Hushbaktova, Z.A...... 160 Koneva, Ekaterina A...... 127 Koptina, Anna V...... 129 I Korchagina, Dina V...... 105, 127, 185, 210 Ibragimova, Aliya Sh...... 52 Korobicina, Elena V...... 165 Il’ina, Irina V...... 70, 105 Korolev, K.G...... 81 Isaev, I.M...... 62, 107 Korolyuk, Elena A...... 187 Isaev, M.I...... 62, 107 Korsunovskaia, O.S...... 193 Isaev, Viacheslav A...... 14 Korulkin, D.U...... 36 Iskenderov, D.A...... 107 Koval, V.S...... 3 Islamova, Jannat I...... 109 Kozhamkulova, Zhanar A...... 148, 240 Itzhanova, Khorlan I...... 95 Kozlov, Nikolas G...... 112, 114 Ivanov, Alexander A...... 234 Kozlov, V.E...... 3 Ivanov, Ivan S...... 110 Kravchenko, Anna O...... 18 Ivasenko, Svetlana A...... 95, 117 Kuandykova, A.Zh...... 208 Kuchin, Alexander V...... 239 J Kudaibergenova, B.M...... 116 Jalmahkanbetova, Roza I...... 111 Kukina, Tatyana P...... 20, 33, 73, 130 Janar, Jenis ...... 31 Kulmukhametova, Z.G...... 125 Javadov, A.S...... 193 Kuzmich, Alexandra S...... 39 Jeon, Je Seung ...... 224 Kuznetsov, Boris N...... 123, 132, 134, 206

162 Authors Index

Kuznetsova, Svetlana A...... 132, 134, 150 N Nabiev, A.N...... 159, 160, 162 L Narchuganov, Anton N...... 37 Laev, Sergey S...... 28, 97 Nasrullaev, A.O...... 5 Lafont, Rene...... 242 Naydenko, Ekaterina S...... 236 Levdansky, Alexander V...... 134 Nechaev, L.V...... 48 Levdansky, Vladimir A...... 134 Nechepurenko, Ivan V...... 163 Lipeeva, Alla V...... 136 Nesterovich, Sergey L...... 165 Litvinenko, Y.A...... 138 Nikitina, Valentina E...... 220 Lobov, A.N...... 152 Nishanbayev, S.Z...... 143, 215 Logutenko, Olga A...... 236 Novikova, O.V...... 89 Logvinov, Sergey V...... 239 Nurmaganbetov, Zhangeldy S...... 166 Lomovsky, I.O...... 139 Lomovsky, Oleg I.35, 81, 100, 139, 144, O 191, 234 Odinokov, Victor N...... 52 Luzina, Olga A...... 217 Oh, Jae Keun...... 38 Ly, Bui Minh...... 229 Omyrzakov, M.T...... 116 Lyakhov, N.Z...... 231 Os’kina, Irina A...... 168 Lytova, Lydmila V...... 14 Osipova, Svetlana O...... 109 Otaeva, Sh.A...... 198, 201 M Ozek, G...... 72 Mahmutova, Almagul...... 194 Mainagashev, Iliya Ya...... 210 P Makarova, Darya L...... 141 Pan, Cheol-Ho...... 24 Mal’tseva, E.V...... 48 Pankratov, Alexei N...... 220 Malykhin, Eugeny V...... 33 Pankrushina, Natalia A...... 43, 170 Mamatkulova, N.M...... 143, 205, 215 Papisova, A.I...... 193 Mamylov, Serguey G...... 144 Pelageev, Dmitry N...... 39 Mansurov, Zh.A...... 219 Petrenko, Natalya I...... 68 Mansurov, Zulkhair A...... 58 Pham, Thinh D...... 229 Matistov, N.V...... 145, 147 Plotnikov, Mark B...... 110, 176, 239 Maximenko, Anastasia V...... 9 Plotnikova, Tatyana M...... 67 Mayeva, Kseniya...... 148 Politov, A.A...... 100, 172 Melikuziev, F.A...... 143 Polovinka, Marina P...... 163, 217 Mikhailapov, I.V...... 3 Polyarush, Svetlana V...... 41 Mikhailenko, M.A...... 150 Polyudina, Revmira I...... 174 Mikhailova, L.R...... 16 Popov, Sergey A...... 33 Minilbaeva, A.V...... 152 Popova, Elena V...... 176 Mironov, Maksim E...... 154 Potapov, Dmitry A...... 174 Mirzaev, Yu.R...... 156 Pozdeeva, Alexandra V...... 185 Misawa, Norihiko...... 23 Praliyev, Kaldybay D...... 56 Mitasov, M.M...... 3 Prosenko, Aleksandr E...... 80, 165 Morozov, S.V...... 3, 83 Pylina, Yana I...... 178 Mullabaeva, Z.U...... 200, 201 Mungunnaran, D...... 29 R Murzakhmetova, Maira K...... 58 Radwan, Mohamed M...... 240 Mussina, A.E...... 158 Rakhadilova, Saniya N...... 179 Muzichkina, R.A...... 36, 138, 158 Rakhimov, Sh.B...... 180 Rakhimova, Bibigul B...... 95, 117, 182 Rakhmadieva, S.B...... 72

163 Authors Index

Ramazanov, N.Sh...... 184 Skvortsova, Galina P...... 132, 134 Rasulova, H.A...... 122 Sokolov, Dmitry N...... 217 Revutsky, Alfred L...... 85 Sokolova, Ekaterina V...... 54 Rodionov, Alexander V...... 203 Sokolova, I.V...... 48 Rogachev, Artem D...... 185 Soloshenko, Vladimir A...... 144 Rogachev, Victor A...... 144 Sorokina, I.V...... 46 Romanenko, Elena P...... 187 Sosnin, E.A...... 48 Romanov, Vasiliy E...... 189 Sostina, Tatiana L...... 123 Ross, Samir A...... 240 Stolyarova, O.V...... 125 Rozhanskaya, Olga A...... 174, 191 Stonik, Valentin A...... 25 Rudenskaya, Galina N...... 14, 193 Sultanova, N.A...... 218, 219 Rudenskaya, Ylia A...... 14, 193 Syrov, Vladimir N...... 109, 160, 205 Ryabchikova, E.I...... 81 Ryazantsev, Oleg G...... 194 T Tashmatov, Z.O...... 94 S Tchaikovskaya, O.N...... 48 Sadikov, A.Z...... 196, 198, 200, 201 Temurova, Sh.A...... 92 Sadyrbekov, Daniyar T...... 194 Tikhonov, Alexey Ya...... 168 Sagdullaev, B.T...... 119, 160 Tikhonova, Elena V...... 194 Sagdullaev, Sh.Sh...... 196, 198, 200, 201 Timakova, Evgeniya V...... 236 Salakhutdinov, Nariman F.28, 70, 97, 105, Tkachenko, Kirill G...... 203 127, 163, 185, 210, 217 Tkachev, Alexey V...... 85, 87, 141, 187 Salnikov, Georgi E...... 105 Tolstikov, Genrikh A.29, 46, 68, 75, 120, Sandanov, Denis V...... 43, 170 136, 154, 163, 166, 185, 189, 210 Sapargalieva, Gaukhar S...... 58 Tolstikova, T.G...... 46 Sergeeva, Olga V...... 176 Tsivileva, Olga M...... 220 Sergeyev, Roman V...... 202 Tsypysheva, I.P...... 152 Shadrin, Dmitry M...... 203 Turdibaev, Zh.E...... 5 Shaimerdenova, Zhanar R...... 182 Turdybekov, Koblandy M...... 194 Shakhidoyatov, Khusnutdin M.5, 75, 143, Turmukhambetov, Aibek Zh...... 166 205, 215 Tuvshintugs, A...... 29 Shakhtshneider, T.P...... 150 Shakirov, R.Sh...... 63, 122, 156, 162 U Shang, Ya Fang ...... 222, 223 Udalova, Tatiana A...... 236 Sharypov, Viktor I...... 206 Um, Byung-Hun6, 170, 222, 223, 224, 225 Shaushekov, Z.K...... 208 Umbetova, A.K...... 218 Sheremet, Olga P...... 104 Ushakova, Yevgeniya S...... 226 Shernyukov, Andrew V...... 210 Sheveleva, Nadezhda N...... 202 V Shin, Kwang Soon...... 54 Vaganova, Tamara A...... 33 Shirshova, T.I...... 145, 147 Valiev, N.V...... 196, 198 Shmidt, Emma N...... 130, 212 Vasil’eva, Natalia V...... 67 Shubina, Larisa K...... 25 Vasiljev, Vladimir G...... 65, 170 Shulgau, Zarina T...... 214 Vereshchagin, N.A...... 50, 227 Shults, Elvira E.29, 45, 68, 75, 120, 136, Veskina, Natalya A...... 52 154, 166, 189 Vinogradova, V.I...... 180 Shurgin, Alexey I...... 129, 202 Vishchuk, Olesya S...... 229 Siddikov, D.R...... 215 Volcho, Konstantin P...... 70, 105, 127 Sidekhmenova, AnastasiaV...... 110 Volodin, Vladimir V...... 53, 178, 203 Sizova, Natalia V...... 216 Volodina, Svetlana O...... 53, 178, 203

164 Authors Index

Vorsina, I.A...... 231 Yusupova, Elvira G...... 238 Vyalkov, A.I...... 83 Z Y Zhabaeva, Anar N...... 95 Yakovleva, Irina M...... 18 Zhang, Zhi-Qiang...... 24 Yermak, Irina M...... 18, 54 Zhantiev, R.D...... 193 Yevtukhova, Larisa A...... 77 Zhdankina, Anna A...... 239 You, Sang Guan ...... 233 Zhumagalieva, Sh.N...... 116 Yu, Valentina K...... 56 Zhusupova, Aizhan I...... 179 Yudina, Natalya V...... 48, 234 Zhusupova, Galiya E...... 58, 148, 219, 240 Yukhin, Yuri M...... 236 Zibareva, Larisa ...... 242 Yunusov, M.S...... 152 Zvyagintseva, Tatyana N...... 229 Yusupov, O.Sh...... 119

165

2nd Annual Russian-Korean Conference “Current issues of natural products chemistry and biotechnology”

Book of abstracts

Подписано к печати 27.02.2010. Формат бумаги 6084/8. Тираж 160 экз.

Отпечатано на полиграфическом участке Новосибирского института органической химии СО РАН 630090, Новосибирск, пр. Академика Лаврентьева, 9