DOCSLIB.ORG

FROM Aristolochia Fimbriata, a BASAL ANGIOSPERM

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

SEQUENCING AND ANALYSIS OF A cDNA ENCODING A PUTATIVE COCLAURINE N-METHYLTRANSFERASE (CNMT) FROM Aristolochia fimbriata, A BASAL ANGIOSPERM DISSERTATION Submitted To THE FACULTY OF LIFE & ENVIRONMENTAL SCIENCES UNIVERSITY OF PESHAWAR In fulfillment for the degree Of DOCTOR OF PHILOSOPHY by ROSHAN ALI THE CENTER OF BIOTECHNOLOGY AND MICROBIOLOGY UNIVERSITY OF PESHAWAR 2010 1 CENTER OF BIOTECHNOLOGY AND MICROBIOLOGY UNIVERSITY OF PESHAWAR SEMESTER 2010 This Dissertation is Submitted By ROSHAN ALI Titled SEQUENCING AND ANALYSIS OF A cDNA ENCODING A PUTATIVE COCLAURINE N-METHYLTRANSFERASE (CNMT) FROM Aristolochia fimbriata, A BASAL ANGIOSPERM In fulfillment of the requirements for the degree Of DOCTOR OF PHILOSOPHY In BIOTECHNOLOGY 2 APPROVAL The dissertation of Mr. Roshan Ali is approved SUPERVISOR: Dr. Ghosia Lutfullah Signature:______________ Professor, Date:__________________ Center of Biotechnology & Microbiology, University of Peshawar. EXAMINER: Signature:_______________ Date:___________________ 3 4 In the name of Allah, the Beneficent, the Merciful 5 Graphical Abstract of the dissertation: Image sources are: (http://1.bp.blogspot.com/_vKxDQBwCntA/S-DMy0e3g VI/AAAAAAAAAJo/e2ickYGXEas/s1600/2516243964_5e41ba8d14.jpg), (http://images19.fotki.com/v27/photos/6/642761/4094024/Aristolochiafimbriat a37-vi.jpg?1182176542), (http://www.scripps.edu/chem/wong/PIX/riboso me.jpg) and this dissertation. 6 TABLE OF CONTENTS Title Page ACKNOWLEDGEMENTS I ABSTRACT V LIST OF ABBREVIATIONS VII LIST OF FIGURES IX LIST OF TABLES XII LIST OF APPENDICES XIII 1. INTRODUCTION 1 1.1. S-adenosyl-L-Methionine-Dependent Methyltransferases 3 1.2. S-Adenosyl-L-Methionine (SAM or AdoMet) 4 1.3. Genes of Bezylisoquinoline Alkaloid Pathway 4 1.4. Alkaloids 6 1.4.1. Types of Alkaloids 8 1.4.2. Benzilisoquinoline Alkaloids (BIAs) 8 1.4.3. BIA Producing Pathway 10 1.4.4. Common Pathway 10 1.4.5. Sanguinarine Pathway 13 1.4.6. Morphine Pathway 14 1.4.7. Medicinal Value of Alkaloids 14 1.5. Aristolochia 20 1.5.1. Medicinal Value of Aristolochia 22 1.6. Aim of the Present Study 24 2. MATERIALS AND METHODS 27 I 2.1. Tissue Source 27 2.2. RNA Extraction 27 2.2.1. Ambion Method 27 2.2.2. Modified CTAB Method 28 2.3. Quality and Purity of the Total RNA 28 2.4. Messenger RNA (mRNA) Isolation 28 2.5. Unigene Selection from A. fimbriata EST Database 29 2.5.1. Papaver somniferum CNMT Gene Sequence Information 29 2.5.2. Tribe Identification of the P. somniferum CNMT Gene and Unigene 29 Selection 2.6. Analysis and Amplification of Gene Expression by Reverse Transcriptase RT-PCR 30 2.6.1. Polymerase Chain Reaction 30 2.6.2. Gene Cloning 31 2.7. Gene Identification and Prediction of the Function of its Putative Protein 32 2.7.1. Similarity Searches 32 2.7.2. Sequence Translation 33 2.7.3. Protein Parameters 33 2.7.4. ModBase Search 33 2.7.5. Function Predictions 34 2.7.6. Secondary Structure Prediction 36 2.8. Homology Model Building 37 2.8.1. Template Identification 37 2.8.2. Target-Template Alignment 38 2.8.3. Model Generation 38 II 2.8.4. Best Structure Selection and its Evaluation 39 2.9. Active Site Identification 40 2.10. Docking 40 2.11. Analysis of Ligand and Substrate Binding Interactions with the Active Site 42 Residues 2.12. Phylogenetic Analysis 43 3. RESULTS AND DISCUSSION 45 3.1. Qualitative Assessment of Extracted Total RNA 45 3.2. PCR and Gel Extraction 51 3.3. Nucleotide Sequence Chromatogram Analysis 54 3.4. Tribe Identification 58 3.5. Identification of the Gene and Prediction of the Putative Function of its 68 Protein 3.5.1. Pairwise Sequence Alignment 68 3.5.2. BLASTn, BLASTx and tBLASTx NCBI Similarity Searches 70 3.5.3. Sequence Translation and BLASTp Search 77 3.5.4. Protein Parameters 83 3.5.5. Protein Properties and Functional Analysis 86 3.5.6. ModBase Search 91 3.5.7. Conserved Domain Database Predictions 93 3.6. Homology Model Building 93 3.6.1. Template Identification 93 3.6.2. Secondary Structure Analysis 100 3.6.3. Model Generation 104 III 3.6.4. Models Evaluation and Selection of the Best Model 106 3.6.5. PROCHECK and ProSA Results 106 3.6.6. Z-Score Values and Their Comparison 113 3.6.7. Ramachandran Plot 119 3.6.8. Superimposition 124 3.7. 3D Structure of the Model 132 3.8. Comparison with Theoretical Models of Other CNMTs 136 3.9. Binding Site 146 3.9.1. Active Site Identification 146 3.9.2. A. fimbriata Putative CNMT Model Active Site Residues as Calculated by Ligplot 151 3.9.3. SAM Binding Site 155 3.9.4. Substrate Binding Pocket 160 3.10. Putative Reaction Mechanism 162 3.11. Phylogenetic Analysis 169 4. CONCLUSIONS 179 5. APPENDIX 182 6. REFERENCES 184 IV ACKNOWLEDGEMENTS To begin with, all praise and thanks to Allah, God Almighty, most Beneficent and Merciful for giving me tremendous courage and power to complete this research and dissertation work. May Allah shower his blessings on Muhammad (SAWWS) and his Ahlulbait (AS), who always have been the sources of knowledge and guidance for humanity. I am greatly thankful to my supervisor Prof. Dr. Ghosia Lutfullah, Professor at the Center of Biotechnology and Microbiology, University of Peshawar, Pakistan for her great help, nice advices, guidance, and encouragement. I am happy to acknowledge my deep sense of gratitude to the Director, Center of Biotechnology and Microbiology Prof. Dr. Bashir Ahmad for his great help, inspiring encouragement and overwhelming support. I wish to express my deep gratitude to my ever smiling Professor Dr. Claude dePamphilis for his excellent guidance, constant encouragement and affectionate patronage throughout the experimental work conducted at dePamphilis laboratory, Department of Biology, The Huck Institute of Life Sciences, University Park, The Pennsylvania State University, USA under his kind supervision. The experimental work embodied in this dissertation was carried out at his laboratory. I am also thankful to him for providing me with all the facilities in the laboratory and financial support. This work would not have been possible without the fellowship award from the Higher Education Commission, Pakistan for which I am thankful to the Government of I Pakistan and particularly grateful to Dr. Atta-u-Rehman who devised this fellowship program. I wish to express my special thanks to Dr. Sayed Awlad Hussain and Dr. Sayed Mussarat Hussain for their sincere guidance, valuable suggestions, inspiring encouragement, fruitful discussion, nice advices and kind prayers. I would like to pay my special regards to their family, Baba buzargawar and their forefathers. I wish my gratitude to the research fellows Kerr Wall, Paula Ralph, Lena Landherr and Laura Warg of USA, Nadira Naznin and Bindhu Abinah of India, Yan Zhang, Yuannian Jiao of China, Norman Wickett of Canada in the dePamphilis laboratory for their great help, cooperation, sincere wishes, invaluable assistance and for their nice company during my stay in USA. I am also thankful to Dr. Ahmad, Zaid, Najeem, Khalid, Wael, Jouke Postma, Maria Postma and their daughter Leizbeth for their beautiful company during my stay in USA. My expressions of gratitude and sincere thanks are due to Mr. Zahid Khan, Mr. Abid Ali Khan, Aziz-ur-Rehman, Ashfaq Ahmad, Sadiq Azam, Ibrar Khan, Sajid Ali, Johar Jameel, Riaz Khan and other research fellows who helped me immensely with their fruitful discussions, valuable suggestions and continuous cooperation. I am thankful to Faculty members Dr. Nafees Bacha, Mr. Akhtar Hussain, Mr. Noor Muhammad, Mr. Jamshid Ahmad, Mr. Abu Nasar Sidiqui, Mr. Momin Khan and Mr. Khalid Javed lecturers at the Center of Biotechnology and Microbiology, University of Peshawar, for their sincere wishes and help during my course work. II Great contribution has been made in polishing my dissertation by Dr. Claude, Paula Ralph, Lena Landherr, Yuannian Jiao. I would convey my thanks to Prof. Dr. S. Mustafa and Dr. Rasool Khan for their guidance, encouragement and cooperation. I am also thankful to all my friends Shuja-u-Din, Aftab Khan, Ashiq Hussain, Nabi Hassan, Muzamil Hussain, Mudasir Hussain, Wahab Ali, S. Mahboob Ali Shah, Izhar Ali, Anwar Ali, Iqbal Hussain and Javed Hussain for their nice company, prayers and encouragement. I wish to express my special thanks to Mr. Javid Hussain and Mr. Shafaat Hussain for their help, cooperation, encouragement, nice company and financial support. I am thankful to all my relatives Noor Alam Jan, Jabir Hussain, Muhammad Ayub, Aftab Hussain, Kamal Hussain, Javid Hussain, Amjad Hussain, Muhammad Ibrahim, Khalid Hussain, Wajid Hussain, Kiramat Hussain, Amir Hussain, Abdullah Jan, Faras Ali, Asad Ali, Tanvir Hussain, Mehdi Hussain, Muhammad Zulqarnain, Hadi Hussain, Muhammad Saki, Qamar Abbas, Ansar Hussain, Ihtisham, Basit Ali, Hassanain, Awais, Nishat Hassan, Waqar Hussain, Anis, Mahmood Ali, Maroof Ali, Ashfaq Hussain, Banaras Hussain, Qaim Hussain, Khatib Hussain and Komail Hussain for their kind prayers. Finally I would like to forward a special word of gratitude to my very supportive mother, my father, Muslim Jan, and my sisters for their enormous cooperation throughout my life, especially my mother whose love, prayers and moral support enabled me to continue with perseverance. Roshan Ali III ABSTRACT IV ABSTRACT Alkaloids are produced in plants through various pathways involving several enzymes that lead to diverse alkaloids. One of the most important alkaloid biosynthetic enzymes is coclaurine N-methyltransferase (CNMT) which is an S-adenosyl-L- methionine-dependent methyltransferase (SAM-MTase). SAM-MTases utilize S- adenosyl-L-methionine (SAM) as a cofactor to methylate other molecules. CNMT catalyzes the methylation of coclaurine. Crystal structures of more than hundred SAM-MTases have been investigated. Several O-methyltransferases have been characterized at the molecular as well as structural levels, but there have been very few molecular studies of N-methyltransferases especially about CNMTs.
Recommended publications
  • 1. Introduction

    1. Introduction

    Introduction 1. Introduction 1.1. Alkaloids The term alkaloid is derived from Arabic word al-qali, the plant from which “soda” was first obtained (Kutchan, 1995). Alkaloids are a group of naturally occurring low-molecular weight nitrogenous compounds found in about 20% of plant species. The majority of alkaloids in plants are derived from the amino acids tyrosine, tryptophan and phenylalanine. They are often basic and contain nitrogen in a heterocyclic ring. The classification of alkaloids is based on their carbon-nitrogen skeletons; common alkaloid ring structures include the pyridines, pyrroles, indoles, pyrrolidines, isoquinolines and piperidines (Petterson et al., 1991; Bennett et al., 1994). In nature, plant alkaloids are mainly involved in plant defense against herbivores and pathogens. Many of these compounds have biological activity which makes them suitable for use as stimulants (nicotine, caffeine), pharmaceuticals (vinblastine), narcotics (cocaine, morphine) and poisons (tubocurarine). The discovery of morphine by the German pharmacist Friedrich W. Sertürner in 1806 began the field of plant alkaloid biochemistry. However, the structure of morphine was not determined until 1952 due to its stereochemical complexity. Major technical advances occurred in this field allowing for the elucidation of selected alkaloid biosynthetic pathways. Among these were the introduction of radiolabeled precursors in the 1950s and the establishment in the 1970s of plant cell suspension cultures as an abundant source of enzymes that could be isolated, purified and characterized. Finally, the introduction of molecular techniques has made possible the isolation of genes involved in alkaloid secondary pathways (Croteau et al., 2000; Facchini, 2001). 1.1.1. Benzylisoquinoline alkaloids Isoquinoline alkaloids represent a large and varied group of physiologically active natural products.
  • Background Document: Roc: Aristolochic Acids ; 2010

    Background Document: Roc: Aristolochic Acids ; 2010

    FINAL Report on Carcinogens Background Document for Aristolochic Acids September 2, 2008 U.S. Department of Health and Human Services Public Health Services National Toxicology Program Research Triangle Park, NC 27709 This Page Intentionally Left Blank RoC Background Document for Aristolochic Acids FOREWORD 1 The Report on Carcinogens (RoC) is prepared in response to Section 301 of the Public 2 Health Service Act as amended. The RoC contains a list of identified substances (i) that 3 either are known to be human carcinogens or are reasonably be anticipated to be human 4 carcinogens and (ii) to which a significant number of persons residing in the United 5 States are exposed. The Secretary, Department of Health and Human Services (HHS), has 6 delegated responsibility for preparation of the RoC to the National Toxicology Program 7 (NTP), which prepares the report with assistance from other Federal health and 8 regulatory agencies and nongovernmental institutions. 9 Nominations for (1) listing a new substance, (2) reclassifying the listing status for a 10 substance already listed, or (3) removing a substance already listed in the RoC are 11 reviewed in a multi-step, scientific review process with multiple opportunities for public 12 comment. The scientific peer-review groups evaluate and make independent 13 recommendations for each nomination according to specific RoC listing criteria. This 14 background document was prepared to assist in the review of aristolochic acids. The 15 scientific information used to prepare Sections 3 through 5 of this document must come 16 from publicly available, peer-reviewed sources. Information in Sections 1 and 2, 17 including chemical and physical properties, analytical methods, production, use, and 18 occurrence may come from published and/or unpublished sources.
  • Aristolochic Acid-Induced Nephrotoxicity: Molecular Mechanisms and Potential Protective Approaches

    Aristolochic Acid-Induced Nephrotoxicity: Molecular Mechanisms and Potential Protective Approaches

    International Journal of Molecular Sciences Review Aristolochic Acid-Induced Nephrotoxicity: Molecular Mechanisms and Potential Protective Approaches Etienne Empweb Anger, Feng Yu and Ji Li * Department of Clinical Pharmacy, School of Basic Medical Sciences and Clinical Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (E.E.A.); [email protected] (F.Y.) * Correspondence: [email protected]; Tel.: +86-139-5188-1242 Received: 25 November 2019; Accepted: 5 February 2020; Published: 10 February 2020 Abstract: Aristolochic acid (AA) is a generic term that describes a group of structurally related compounds found in the Aristolochiaceae plants family. These plants have been used for decades to treat various diseases. However, the consumption of products derived from plants containing AA has been associated with the development of nephropathy and carcinoma, mainly the upper urothelial carcinoma (UUC). AA has been identified as the causative agent of these pathologies. Several studies on mechanisms of action of AA nephrotoxicity have been conducted, but the comprehensive mechanisms of AA-induced nephrotoxicity and carcinogenesis have not yet fully been elucidated, and therapeutic measures are therefore limited. This review aimed to summarize the molecular mechanisms underlying AA-induced nephrotoxicity with an emphasis on its enzymatic bioactivation, and to discuss some agents and their modes of action to reduce AA nephrotoxicity. By addressing these two aspects, including mechanisms of action of AA nephrotoxicity and protective approaches against the latter, and especially by covering the whole range of these protective agents, this review provides an overview on AA nephrotoxicity. It also reports new knowledge on mechanisms of AA-mediated nephrotoxicity recently published in the literature and provides suggestions for future studies.
  • Aristolochic Acids Tract Urothelial Cancer Had an Unusually High Incidence of Urinary- Bladder Urothelial Cancer

    Aristolochic Acids Tract Urothelial Cancer Had an Unusually High Incidence of Urinary- Bladder Urothelial Cancer

    Report on Carcinogens, Fourteenth Edition For Table of Contents, see home page: http://ntp.niehs.nih.gov/go/roc Aristolochic Acids tract urothelial cancer had an unusually high incidence of urinary- bladder urothelial cancer. CAS No.: none assigned Additional case reports and clinical investigations of urothelial Known to be human carcinogens cancer in AAN patients outside of Belgium support the conclusion that aristolochic acids are carcinogenic (NTP 2008). The clinical stud- First listed in the Twelfth Report on Carcinogens (2011) ies found significantly increased risks of transitional-cell carcinoma Carcinogenicity of the urinary bladder and upper urinary tract among Chinese renal- transplant or dialysis patients who had consumed Chinese herbs or Aristolochic acids are known to be human carcinogens based on drugs containing aristolochic acids, using non-exposed patients as sufficient evidence of carcinogenicity from studies in humans and the reference population (Li et al. 2005, 2008). supporting data on mechanisms of carcinogenesis. Evidence of car- Molecular studies suggest that exposure to aristolochic acids is cinogenicity from studies in experimental animals supports the find- also a risk factor for Balkan endemic nephropathy (BEN) and up- ings in humans. per-urinary-tract urothelial cancer associated with BEN (Grollman et al. 2007). BEN is a chronic tubulointerstitial disease of the kidney, Cancer Studies in Humans endemic to Serbia, Bosnia, Croatia, Bulgaria, and Romania, that has The evidence for carcinogenicity in humans is based on (1) findings morphology and clinical features similar to those of AAN. It has been of high rates of urothelial cancer, primarily of the upper urinary tract, suggested that exposure to aristolochic acids results from consump- among individuals with renal disease who had consumed botanical tion of wheat contaminated with seeds of Aristolochia clematitis (Ivic products containing aristolochic acids and (2) mechanistic studies 1970, Hranjec et al.
  • Nephrotoxicity and Chinese Herbal Medicine

    Nephrotoxicity and Chinese Herbal Medicine

    Nephrotoxicity and Chinese Herbal Medicine Bo Yang,1,2 Yun Xie,2,3 Maojuan Guo,4 Mitchell H. Rosner,5 Hongtao Yang,1 and Claudio Ronco2,6 Abstract Chinese herbal medicine has been practicedfor the prevention, treatment, andcure of diseases forthousands of years. Herbal medicine involves the use of natural compounds, which have relatively complex active ingredients with varying degrees of side effects. Some of these herbal medicines are known to cause nephrotoxicity, which can be overlooked by physicians and patients due to the belief that herbal medications are innocuous. Some of the 1Department of nephrotoxic components from herbs are aristolochic acids and other plant alkaloids. In addition, anthraquinones, Nephrology, First flavonoids, and glycosides from herbs also are known to cause kidney toxicity. The kidney manifestations of Teaching Hospital of nephrotoxicity associated with herbal medicine include acute kidney injury, CKD, nephrolithiasis, rhabdomyolysis, Tianjin University of Traditional Chinese Fanconi syndrome, and urothelial carcinoma. Several factors contribute to the nephrotoxicity of herbal medicines, Medicine, Tianjin, including the intrinsic toxicity of herbs, incorrect processing or storage, adulteration, contamination by heavy China; 2International metals, incorrect dosing, and interactions between herbal medicines and medications. The exact incidence of kidney Renal Research injury due to nephrotoxic herbal medicine is not known. However, clinicians should consider herbal medicine use in Institute of Vicenza and 6Department of patients with unexplained AKI or progressive CKD. In addition, exposure to herbal medicine containing aristolochic Nephrology, Dialysis acid may increase risk for future uroepithelial cancers, and patients require appropriate postexposure screening. and Transplantation, Clin J Am Soc Nephrol 13: 1605–1611, 2018.
  • United States Patent [191 [11] Patent Number: 4,758,639 Koyanagi Et Al

    United States Patent [191 [11] Patent Number: 4,758,639 Koyanagi Et Al

    United States Patent [191 [11] Patent Number: 4,758,639 Koyanagi et al. [45] Date of Patent: Jul. 19, 1988 [54] PROCESS FOR PRODUCTION OF VINYL [56] References Cited POLYMER . U.S. PATENT DOCUMENTS [75] Inventors: Shunichi Koyanagi, Yokohama; 3,923,765 12/1975 Goetze et al. ....................... .. 526/62 Hajime Kutamura, Ichihara; 4,049,895 9/1977 McOnie et al. ..... .. 526/62 Toshihide Shimizu; Ichiro Kaneko, 4,539,230 9/ 1985 Shimizu et al. ................. .. 526/62 X both of Ibaraki, all of Japan Primary Examiner-Joseph L. Schofer Assistant Examiner-F. M. Teskin [73] Assignee: Shin-Etsu Chemical Co., Ltd., Tokyo, Attorney, Agent, or Firm—0blon, Fisher, Spivak, Japan McClelland & Maier [21] Appl. No.: 94,020 [57] ABSTRACT [22] Filed: Sep. 3, 1987 A process for production of a vinyl polymer by suspen sion polymerization or emulsion polymerization of at Related US. Application Data least one kind of vinyl monomer in an aqueous medium is disclosed. [63] Continuation of Ser. No. 765,803, Aug. 15, 1985, aban In this process, the polymerization is carried out in a cloned. polymerizer, the inner wall surface and portions of the [30] Foreign Application Priority Data auxiliary equipment thereof which may come into contact with the monomer during polymerization hav Aug. 17, 1984 [JP] Japan ........ .. .. 59471045 ing a surface roughness of not greater than 5 pm and Aug. 17, 1984 [JP] Japan .............................. .. 59-171046 being previously coated with a scaling preventive com [51] Int. Cl.‘ ......................... .. C08F 2/18; C08F 2/22; prising at least one selected from dyes, pigments and CO8F 2/44 aromatic or heterocyclic compounds having at least 5 [52] US.
  • Evaluation of the Toxicity Potential of Acute and Sub-Acute Exposure to the Aqueous Root Extract of Aristolochia Ringens Vahl

    Evaluation of the Toxicity Potential of Acute and Sub-Acute Exposure to the Aqueous Root Extract of Aristolochia Ringens Vahl

    Journal of Ethnopharmacology 244 (2019) 112150 Contents lists available at ScienceDirect Journal of Ethnopharmacology journal homepage: www.elsevier.com/locate/jethpharm Evaluation of the toxicity potential of acute and sub-acute exposure to the aqueous root extract of Aristolochia ringens Vahl. (Aristolochiaceae) T Flora R. Aigbea,*, Oluwatoyin M. Sofidiyab, Ayorinde B. Jamesc, Abimbola A. Sowemimob, Olanrewaju K. Akinderea, Miriam O. Aliua, Alimat A. Dosunmua, Micah C. Chijiokea, Olufunmilayo O. Adeyemia a Department of Pharmacology, Therapeutics and Toxicology, Faculty of Basic Medical Sciences, University of Lagos, P.M.B. 12003, Idi-Araba, Surulere, Lagos, Nigeria b Department of Pharmacognosy, Faculty of Pharmacy, College of Medicine, University of Lagos, P.M.B. 12003, Idi-Araba, Surulere, Lagos, Nigeria c Department of Biochemistry & Nutrition, Nigerian Institute of Medical Research, Yaba, Lagos, Nigeria. ARTICLE INFO ABSTRACT Keywords: Ethnopharmacological relevance: Aristolochia ringens Vahl. (Aristolochiaceae) is used traditionally in Nigeria for Aristolochia ringens managing a number of ailments including gastrointestinal disturbances, rheumatoid arthritis, pile, insomnia, Aristolochic acid I oedema, and snake bite venom. Some studies in our laboratory have demonstrated a scientific justification for Brine shrimp some of such uses. This study aims at investigating the toxicological actions of the aqueous root extract of Rodents Aristolochia ringens (AR). Sub-acute toxicity Materials and methods: Brine shrimp lethality assay was carried out using 10, 100 and 1000 μg/ml of the extract. Oral and intraperitoneal acute toxicity tests were carried out using mice. The effect of sub-acute (30 days) repeated oral exposure to the extract at 10, 50 and 250 mg/kg in rats was also evaluated via weekly assessments of body weights and general observations as well as end of exposure haematological, biochemical and histo- logical examinations of blood and tissue samples of treated rats.
  • Influence of Different Elicitors on BIA Production in Macleaya Cordata

    Influence of Different Elicitors on BIA Production in Macleaya Cordata

    www.nature.com/scientificreports OPEN Infuence of diferent elicitors on BIA production in Macleaya cordata Peng Huang1,2,7, Liqiong Xia3,7, Li Zhou1,7, Wei Liu1,4, Peng Wang1, Zhixing Qing5* & Jianguo Zeng1,6* Sanguinarine (SAN) and chelerythrine (CHE) have been widely used as substitutes for antibiotics for decades. For a long time, SAN and CHE have been extracted from mainly Macleaya cordata, a plant species that is a traditional herb in China and belongs to the Papaveraceae family. However, with the sharp increase in demand for SAN and CHE, it is necessary to develop a new method to enhance the supply of raw materials. Here, we used methyl jasmonate (MJ), salicylic acid (SA) and wounding alone and in combination to stimulate aseptic seedlings of M. cordata at 0 h, 24 h, 72 h and 120 h and then compared the diferences in metabolic profles and gene expression. Ultimately, we found that the efect of using MJ alone was the best treatment, with the contents of SAN and CHE increasing by 10- and 14-fold, respectively. However, the increased SAN and CHE contents in response to combined wounding and MJ were less than those for induced by the treatment with MJ alone. Additionally, after MJ treatment, SAN and CHE biosynthetic pathway genes, such as those encoding the protopine 6-hydroxylase and dihydrobenzophenanthridine oxidase enzymes, were highly expressed, which is consistent with the accumulation of SAN and CHE. At the same time, we have also studied the changes in the content of synthetic intermediates of SAN and CHE after elicitor induction.
  • United States Patent (19) 11 Patent Number: 6,123,943 Baba Et Al

    United States Patent (19) 11 Patent Number: 6,123,943 Baba Et Al

    USOO61239.43A United States Patent (19) 11 Patent Number: 6,123,943 Baba et al. (45) Date of Patent: Sep. 26, 2000 54 NF-KBACTIVITY INHIBITOR 8-301761 11/1996 Japan ............................. A61K 31/47 75 Inventors: Masanori Baba, Kagoshima; Minoru OTHER PUBLICATIONS Ono, Tokyo, both of Japan Sato et al. Eur. J. Pharmacol. (1982) 83: 91-95. 73 Assignee: Kaken Shoyaku Co., Ltd., Tokyo, Japan Primary Examiner Jean C. Witz Attorney, Agent, or Firm Sughrue, Mion, Zinn, Macpeak 21 Appl. No.: 09/037,712 & Seas, PLLC 22 Filed: Mar 10, 1998 57 ABSTRACT 30 Foreign Application Priority Data This invention relates to an NF-kB activity inhibitor which contains alkaloids originated from a plant belonging to the Dec. 22, 1997 JP Japan .................................... 9-353879 genus Stephania of the family Menspermaceae, derivatives thereof and Salts thereof, as the active components, to an (51) Int. Cl." ..................................................... A61K 35/78 agent for use in the treatment and prevention of diseases 52 U.S. Cl. ....................... 424/195.1; 514/308; 514/387; upon which the NF-kB activity inhibiting action is effective 514/415 and to an inhibitor of the expression of related genes. Since 58 Field of Search ......................... 424/195.1; 514/387, Said active components exert an action to inhibit transcrip 514/308, 415; 546/140,139; 534/790 tion of DNA having an NF-kB recognition sequence by inhibiting the activity of NF-kB, the drug of the present 56) References Cited invention can inhibit expression of genes of certain Sub U.S. PATENT DOCUMENTS stances Such as cytokines, inflammatory cytokine receptor antagonists, MHC class I, MHC class II, B2 microglobulin, 5,025,020 6/1991 Van Dyke ..............................
  • Chemical Constituents and Pharmacology of the Aristolochia ( 馬兜鈴 Mădōu Ling) Species

    Chemical Constituents and Pharmacology of the Aristolochia ( 馬兜鈴 Mădōu Ling) Species

    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Journal of Traditional and Complementary Medicine Vol. 2, No. 4, pp. 249-266 Copyright © 2011 Committee on Chinese Medicine and Pharmacy, Taiwan. This is an open access article under the CC BY-NC-ND license. :ŽƵƌŶĂůŽĨdƌĂĚŝƚŝŽŶĂůĂŶĚŽŵƉůĞŵĞŶƚĂƌLJDĞĚŝĐŝŶĞ Journal homepagĞŚƩƉ͗ͬͬǁǁǁ͘ũƚĐŵ͘Žƌg Chemical Constituents and Pharmacology of the Aristolochia ( 馬兜鈴 mădōu ling) species Ping-Chung Kuo1, Yue-Chiun Li1, Tian-Shung Wu2,3,4,* 1 Department of Biotechnology, National Formosa University, Yunlin 632, Taiwan, ROC 2 Department of Chemistry, National Cheng Kung University, Tainan 701, Taiwan, ROC 3 Department of Pharmacy, China Medical University, Taichung 404, Taiwan, ROC 4 Chinese Medicine Research and Development Center, China Medical University and Hospital, Taichung 404, Taiwan, ROC Abstract Aristolochia (馬兜鈴 mǎ dōu ling) is an important genus widely cultivated and had long been known for their extensive use in traditional Chinese medicine. The genus has attracted so much great interest because of their numerous biological activity reports and unique constituents, aristolochic acids (AAs). In 2004, we reviewed the metabolites of Aristolochia species which have appeared in the literature, concerning the isolation, structural elucidation, biological activity and literature references. In addition, the nephrotoxicity of aristolochic acids, biosynthetic studies, ecological adaptation, and chemotaxonomy researches were also covered in the past review. In the present manuscript, we wish to review the various physiologically active compounds of different classes reported from Aristolochia species in the period between 2004 and 2011. In regard to the chemical and biological aspects of the constituents from the Aristolochia genus, this review would address the continuous development in the phytochemistry and the therapeutic application of the Aristolochia species.
  • Argemone Ochroleuca: (PAPAVERACEAE), ALKALOID POTENTIAL SOURCE for AGRICULTURAL and MEDICINAL USES †

    Argemone Ochroleuca: (PAPAVERACEAE), ALKALOID POTENTIAL SOURCE for AGRICULTURAL and MEDICINAL USES †

    Tropical and Subtropical Agroecosystems 23 (2020): #31 Hernández-Ruiz et al., 2020 Review [Revisión] Argemone ochroleuca: (PAPAVERACEAE), ALKALOID POTENTIAL SOURCE FOR AGRICULTURAL AND MEDICINAL USES † [Argemone ochroleuca: (PAPAVERACEAE), FUENTE POTENCIAL DE ALCALOIDES PARA LA AGRICULTURA, Y USO MEDICINAL] J. Hernández-Ruiz1, J. Bernal2, J. Gonzales-Castañeda1, J. E. Ruiz-Nieto1 and A. I. Mireles-Arriaga1* 1División de Ciencias de la Vida, Universidad de Guanajuato. Km 9 carretera Irapuato-Silao, Ex Hacienda. El Copal, Irapuato, Guanajuato. 36500 México. Email: [email protected] 2Department of Entomology, Texas A&M University, College Station, TX 77843-247, USA *Corresponding author SUMMARY Background. The genus Argemone contains 24 species, A. ochorleuca is present in national territory and is used in agriculture and traditional medical treatments for various conditions. Results. A. ochorleuca is an herbaceous and/or perennial plant that blooms all year. This plant had the potential as a source of benzyl isoquinoline alkaloids, which are the main bioactive compounds responsible for antibacterial, antifungal properties. However, some of these compounds are associated with toxic effects too. Information about concentrations and parts of the plant it is important for all uses and applications. Implications. The present work summarizes available information on phytochemical and medicinal properties. Conclusion. In A. ochrolecuca, six of the 45 alkaloids reported for the genus Argemone have been studied, dihydro-keleritrin and dihydro-sanguiranine are the most abundant in the seeds and vegetative tissue of the species. The updated information should be useful to guide future research on this plant. Keywords: Alkaloids; papaveraceae; berberine; sanguinarine. RESUMEN Antecedentes. El género Argemone contiene 24 especies, A.
  • Natural Products As Chemopreventive Agents by Potential Inhibition of the Kinase Domain in Erbb Receptors

    Natural Products As Chemopreventive Agents by Potential Inhibition of the Kinase Domain in Erbb Receptors

    Supplementary Materials: Natural Products as Chemopreventive Agents by Potential Inhibition of the Kinase Domain in ErBb Receptors Maria Olivero-Acosta, Wilson Maldonado-Rojas and Jesus Olivero-Verbel Table S1. Protein characterization of human HER Receptor structures downloaded from PDB database. Recept PDB resid Resolut Name Chain Ligand Method or Type Code ues ion Epidermal 1,2,3,4-tetrahydrogen X-ray HER 1 2ITW growth factor A 327 2.88 staurosporine diffraction receptor 2-{2-[4-({5-chloro-6-[3-(trifl Receptor uoromethyl)phenoxy]pyri tyrosine-prot X-ray HER 2 3PP0 A, B 338 din-3-yl}amino)-5h-pyrrolo 2.25 ein kinase diffraction [3,2-d]pyrimidin-5-yl]etho erbb-2 xy}ethanol Receptor tyrosine-prot Phosphoaminophosphonic X-ray HER 3 3LMG A, B 344 2.8 ein kinase acid-adenylate ester diffraction erbb-3 Receptor N-{3-chloro-4-[(3-fluoroben tyrosine-prot zyl)oxy]phenyl}-6-ethylthi X-ray HER 4 2R4B A, B 321 2.4 ein kinase eno[3,2-d]pyrimidin-4-ami diffraction erbb-4 ne Table S2. Results of Multiple Alignment of Sequence Identity (%ID) Performed by SYBYL X-2.0 for Four HER Receptors. Human Her PDB CODE 2ITW 2R4B 3LMG 3PP0 2ITW (HER1) 100.0 80.3 65.9 82.7 2R4B (HER4) 80.3 100 71.7 80.9 3LMG (HER3) 65.9 71.7 100 67.4 3PP0 (HER2) 82.7 80.9 67.4 100 Table S3. Multiple alignment of spatial coordinates for HER receptor pairs (by RMSD) using SYBYL X-2.0. Human Her PDB CODE 2ITW 2R4B 3LMG 3PP0 2ITW (HER1) 0 4.378 4.162 5.682 2R4B (HER4) 4.378 0 2.958 3.31 3LMG (HER3) 4.162 2.958 0 3.656 3PP0 (HER2) 5.682 3.31 3.656 0 Figure S1.