DOCSLIB.ORG

Fungal Mediated Biotransformation of Melengestrol Acetate, and T-Cell Proliferation Inhibitory Activity of Biotransformed Compounds

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Fungal Mediated Biotransformation of Melengestrol Acetate, and T-Cell Proliferation Inhibitory Activity of Biotransformed Compounds Bioorganic Chemistry 104 (2020) 104313 Contents lists available at ScienceDirect Bioorganic Chemistry journal homepage: www.elsevier.com/locate/bioorg Fungal mediated biotransformation of melengestrol acetate, and T-cell proliferation inhibitory activity of biotransformed compounds Saira Javed a, Atia-tul-Wahab b,*, Almas Jabeen b, Shynar Zhumagaliyeva c, Zharylkasyn A. Abilov c, Atta-ur-Rahman a, M. Iqbal Choudhary a,b,c,d,* a H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan b Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan c Al-Farabi Kazakh National University, Department of Chemistry and Chemical Technology, Almaty, Kazakhstan d Department of Chemistry, Faculty of Science and Technology, Universitas Airlangga, Komplek Campus C, Surabaya 60115, Indonesia ARTICLE INFO ABSTRACT Keywords: Glomerella fusaroide, and Rhizopus stolonifer were effectively able to transform the steroidal hormone melen­ Melengestrol acetate gestrol acetate (MGA) (1) into four (4) new metabolites, 17α-acetoxy-11α-hydroxy-6-methyl-16-methyl­ Biotransformation enepregna-4,6-diene-3,20-dione (2), 17α-acetoxy-11α-hydroxy-6-methyl-16-methylenepregna-1,4,6-triene-3,20- T-cell Proliferation dione (3), 17α-acetoxy-6,7α-epoxy-6β-methyl-16-methylenepregna-4,6-diene-3,20-dione (4), and 17α-acetoxy- Anti-inflammatory 11β,15β-dihydroxy-6-methyl-16-methylenepregna-4,6-diene-3,20-dione (5). All these compounds were struc­ turally characterized by different spectroscopic techniques. The objective of the current study was to assess the anti-inflammatory potential of melengestrol acetate (1), and its metabolites 2–5. The metabolites and the sub­ strate were assessed for their inhibitory effects on proliferation of T-cells in vitro. The substrate (IC50 = 2.77 ± 0.08 µM) and its metabolites 2 (IC50 = 2.78 ± 0.07 µM), 4 (IC50 = 2.74 ± 0.1 µM), and 5 (IC50 = < 2 µM) exhibited potent T- cell proliferation inhibitory activities, while compound 3 (IC50 = 29.9 ± 0.09 µM) showed a moderate activity in comparison to the standard prednisolone (IC50 = 9.73 ± 0.08 µM). All the metabolites were found to be non-toxic against 3T3 normal cell line. This study thus identifies some potent compounds active against T-cell proliferation. Their anti-inflammatory potential, therefore, deserves to be further investigated. 1. Introduction effectiveness of the process. Biocatalytic reactions involve different types of reactions, such as hydroxylation, reduction, Michael addition, Among various natural and synthetic classes of chemical compounds, oxidation, esterification, epimerization, reverse aldol reaction, and steroid- based drugs are the most extensively used for various health epoxidation [9–15]. Microbial transformations of steroids on a large disorders due to their contraceptive, anti-cancer, anti-androgenic, anti- scale in the synthesis of drugs and other bioactive analogues have been HIV, anti-tumor, anabolic, anti-bacterial, anti-inflammatory, and pro­ reported earlier [3,16]. Fungal-mediated biotransformation is exten­ gestational properties [1,2]. The synthesis of analogues of steroids sively used for the conversion of steroids, and other organic compounds. through conventional chemical methods is often difficult [3–7]. How­ Structurally modified compounds, obtained through biocatalytic re­ ever, biocatalysts are capable of carrying out regio-, and stereoselective actions, are regio-, and stereo-selective with altered therapeutic index reactions efficiently. [2,3,7,9–12,17–19]. These modified compounds may have higher po­ Microbial transformation is an effective tool to generate different tency, reduced adverse effects, and longer half-lives in blood, as oxygenated analogues of steroids and other natural products [4,7,8]. compared to their parent substances [2,3]. The enzyme cytochrome Biocatalytic reactions are more favorable than chemical synthesis P450 monooxygenases, found specificallyin fungi, is responsible for the because of their eco-friendly nature, mild reaction conditions, and cost- selective hydroxylation of various complex steroids [6,12,18,20]. * Corresponding authors at: Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan (Atia-tul-Wahab); H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan (M. Iqbal Choudhary). E-mail addresses: [email protected] (Atia-tul-Wahab), [email protected] (M.I. Choudhary). https://doi.org/10.1016/j.bioorg.2020.104313 Received 16 July 2020; Received in revised form 17 September 2020; Accepted 20 September 2020 Available online 24 September 2020 0045-2068/© 2020 Elsevier Inc. All rights reserved. S. Javed et al. Bioorganic Chemistry 104 (2020) 104313 Melengestrol acetate (1) (C25H32O4) is an orally active synthetic to obtain brown gummy crude. For fractionation of obtained crude, progestagenic steroid, capable of suppressing estrus (heat) in heifers column chromatography was employed. Different chromatographic [21]. MGA (1) is a synthetic analogue of medroxyprogesterone acetate techniques were used to purify resulting metabolites from the crude (MPA), which also exhibits glucocorticoid activity. Glucocorticoids are extract. generally used against many inflammatory disorders [22,23]. We have previously reported many structural modificationsof steroids to produce 2.4. Fermentation of melengestrol acetate (1) with Glomerella fusaroide analogues with diverse activities. Microbial transformation of MGA (1) was previously reported with Cunninghamella blakesleeana, and a new Melengestrol acetate (1) (1 g) was dissolved in 25 mL of acetone, and oxygenated analogue was obtained. dispersed in 25 Erlenmyer flasks (1 L), each contained 400 mL of In continuation of our research on microbial transformations, MGA autoclaved media cultured with Glomerella fusaroide. All flasks were ◦ (1) was incubated with Glomerella fusaroide, and Rhizopus stolonifer, incubated for 8 days on rotary shaker at 27 C. After complete fermen­ which yielded four new metabolites 2–5. Immunosuppressant activity of tation, reaction was quenched by adding ethyl acetate. Each flask was the resulting metabolites was evaluated through the inhibitory effects on then filtered, and extracted by ethyl acetate 3 times. The collected sol­ the proliferation of T-cells in human blood in vitro. vent layer was evaporated on rotavapor to obtain brown gummy crude material. The crude material was then loaded on a silica gel column for 2. Materials and methods fractionation using solvent gradients of pet. ether, and acetone. Two main fractions were obtained at pet. ether: acetone (70:30), and (67:33). 2.1. Materials. These two fractions were further subjected to recycling HPLC. Metabo­ lites 2 (Rt = 42 min), and 3 (Rt = 36 min) were purified at M-80, Melengestrol acetate (1) was obtained from Haihang Industry Co., methanol–water (70:30). Ltd. Beijing, China. Precoated TLC plates (silica gel, 0.25 mm) were purchased from Merck, Germany. Ceric sulfate was used as a TLC 2.4.1. 17α-Acetoxy-11α-hydroxy-6-methyl-16-methylenepregna-4,6- staining reagent. Fractions were obtained by column chromatography, diene-3,20-dione (2) utilizing silica gel (70–230 mesh, Merck). Purified metabolites were ◦ [ ]25 = + = White solid: M. p.: 222–225 C, α D 7.5 (c 0.01, CHCl3), UV obtained through recycling preparative HPLC [JAI LC-908 W equipped, 1 (MeOH) λmax nm: 225, and 288 nm. IR (CHCl3) Vmax cm : 3433 (OH), with YMC M-80 column (4–5 μm, 20–250 mm i.d.)]. UV light of 254 nm – 1 13 2944 (CH), and 1739 (C–O), H NMR (CD3OD; 400 MHz) Table-1; C was used. JEOL (Japan) JMS-600H mass spectrometer was used to + NMR (CD3OD; 100 MHz) Table-2; EI-MS: m/z 412.3 [M] , 353 (61), 309 + measure the electron impact mass spectra (EI-MS), and high-resolution (42); HREI-MS m/z 412.2250 [M ] (mol. formula, calcd value: mass spectra (HREI-MS) in m/z (rel. %). Bruker Avance NMR spec­ C25H32O5, 412.2251). trometers (Bruker, Switzerland) were used to record 1H NMR spectra at 13 400, and 500 MHz, and C NMR spectra at 100, and 125 MHz in 2.4.2. 17α-Acetoxy-11α-hydroxy-6-methyl-16-methylenepregna-1,4,6- CD3OD, respectively. Melting point of each compound was measured on triene-3,20-dione (3) Buchi-560 (Switzerland). For measuring the optical rotation in meth­ ◦ [ ]25 = + = White solid: M. p.: 238–240 C, α D 5.7 (c 0.008, CHCl3), UV anol, JASCO P-2000 polarimeter (Japan) was used. UV Spectra (in nm) 1 (MeOH) λmax nm: 213, 228, 261, 301 nm. IR (CHCl3) Vmax cm : 3400 were recorded on Hitachi U-3200 spectrophotometer (Japan). FT-IR- – – 1 (OH), 2931 (CH), and 1739 (C–O), and 1650 (C–C), H NMR (CD3OD; 8900, and Bruker VECTOR 22 spectrophotometers (Bruker, France) 13 1 500 MHz) Table-1; C NMR (CD3OD; 125 MHz) Table-2; EI-MS: m/z were used to record IR spectra (cm ). + + 410.3 [M] , 368 (26), 351 (100). HREI-MS m/z 410.2093 [M ] (mol. formula, calcd value: C H O , 410.2075). 2.2. Culture medium. 25 30 5 The growth medium for Glomerella fusaroide was prepared by adding 2.5. Fermentation of melengestrol acetate (1) with Rhizopus stolonifer glucose (100 g), glycerol (100 mL), peptone (50 g), yeast extract (50 g), 1.0 g of MGA (1) dissolved in 25 mL acetone was distributed among KH2PO4 (50 g), and NaCl (50 g) into 10.0 L of distilled H2O. Similarly, the medium for Rhizopus stolonifer (TSY- 0471) was formulated by 27 conical flasks (1 L, each containing 400 mL media with complete growth of Rhizopus stolonifer). All 27 flasks were placed on shaker for mixing following ingredients in 20.0 L of distilled H2O: glucose (200 g), fermentation reaction for 14 days. After 14 days, ethyl acetate was used peptone (100 g), KH2PO4 (100 g), and yeast extract (60 g).
Load more

Recommended publications
  • Record of Reasons for 16-18 November 1999 Meeting
    1 NATIONAL DRUGS AND POISONS SCHEDULE COMMITTEE MEETING 25 – 16-18 November 1999 RECORD OF REASONS FOR AMENDMENT TO THE STANDARD FOR THE UNIFORM SCHEDULING OF DRUGS AND POISONS AGRICULTURAL, VETERINARY AND DOMESTIC CHEMICALS Buprofezin – Schedule required Amendment and Reasons The decision below was based on buprofezin’s toxicological profile, in particular its acute oral toxicity. A cut-off to exempt at a concentration of 40% was agreed because of the reduced oral toxicity at this concentration and the ability of safety directions and warning statements applied through the registration system for agricultural and veterinary chemicals to adequately address the slight eye irritation which was attributed to the product formulation. Schedule 5 – New entry BUPROFEZIN except in preparations containing 40 per cent or less of buprofezin. Copper hydroxide - progression of foreshadowed decision to include copper hydroxide in Schedule 6 with cut-off to Schedule 5 at 50 per cent and to exempt at 12.5 per cent. Amendment and Reasons The amendment below was based on the acute oral toxicity of copper hydroxide, its severe eye irritancy / corrosivity, and the possibility of products containing copper hydroxide being accessible in domestic situations and therefore presenting a risk of accidental ingestion. Schedule 6 – New entry COPPER HYDROXIDE except: (a) when included in Schedule 5; or (b) in preparations containing 12.5 per cent or less of copper hydroxide. Schedule 5 – New entry COPPER HYDROXIDE in preparations containing 50 per cent or less of copper hydroxide except in preparations containing 12.5 per cent or less of copper hydroxide. Cyhexatin - Review of Appendix F warning statements and Appendix J rider.
    [Show full text]
  • Megestrol Acetate/Melengestrol Acetate 2115 Adverse Effects and Precautions Megestrol Acetate Is Also Used in the Treatment of Ano- 16
    Megestrol Acetate/Melengestrol Acetate 2115 Adverse Effects and Precautions Megestrol acetate is also used in the treatment of ano- 16. Mwamburi DM, et al. Comparing megestrol acetate therapy with oxandrolone therapy for HIV-related weight loss: similar As for progestogens in general (see Progesterone, rexia and cachexia (see below) in patients with cancer results in 2 months. Clin Infect Dis 2004; 38: 895–902. p.2125). The weight gain that may occur with meges- or AIDS. The usual dose is 400 to 800 mg daily, as tab- 17. Grunfeld C, et al. Oxandrolone in the treatment of HIV-associ- ated weight loss in men: a randomized, double-blind, placebo- trol acetate appears to be associated with an increased lets or oral suspension. A suspension of megestrol ace- controlled study. J Acquir Immune Defic Syndr 2006; 41: appetite and food intake rather than with fluid reten- tate that has an increased bioavailability is also availa- 304–14. tion. Megestrol acetate may have glucocorticoid ef- ble (Megace ES; Par Pharmaceutical, USA) and is Hot flushes. Megestrol has been used to treat hot flushes in fects when given long term. given in a dose of 625 mg in 5 mL daily for anorexia, women with breast cancer (to avoid the potentially tumour-stim- cachexia, or unexplained significant weight loss in pa- ulating effects of an oestrogen—see Malignant Neoplasms, un- Effects on carbohydrate metabolism. Megestrol therapy der Precautions of HRT, p.2075), as well as in men with hot 1-3 4 tients with AIDS. has been associated with hyperglycaemia or diabetes mellitus flushes after orchidectomy or anti-androgen therapy for prostate in AIDS patients being treated for cachexia.
    [Show full text]
  • University of Groningen Multi-Residue Analysis of Growth Promotors In
    University of Groningen Multi-residue analysis of growth promotors in food-producing animals Koole, Anneke IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 1998 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Koole, A. (1998). Multi-residue analysis of growth promotors in food-producing animals. s.n. Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum. Download date: 25-09-2021 APPENDIX 1 OVERVIEW OF RELEVANT SUBSTANCES This appendix consists of two parts. First, substances that are relevant for the research presented in this thesis are given. For each substance CAS number (CAS), molecular weight (MW), bruto formula (formula) and if available UV maxima and alternative names are given. In addition, pKa values for the ß-agonists are listed, if they were available.
    [Show full text]
  • Jim Lauderdale.Pdf
    Proceedings, Applied Reproductive Strategies in Beef Cattle January 28-29, 2010; San Antonio, TX WHERE WE HAVE BEEN AND WHERE WE ARE TODAY: HISTORY OF THE DEVELOPMENT OF PROTOCOLS FOR BREEDING MANAGEMENT OF CATTLE THROUGH SYNCHRONIZATION OF ESTRUS AND OVULATION J. W. Lauderdale Lauderdale Enterprises, Inc. Augusta, MI 49012 Introduction Reproductive efficiency is one of the most important factors for successful cow-calf enterprises. Certainly, in the absence or reproduction, there is no cow-calf enterprise. During the 1950s frozen bovine semen was developed and AI with progeny tested bulls became recognized as effective to make more rapid genetic progress for milk yield and beef production. During the interval 1950s through 1960s, a major detriment to AI in beef cattle was the requirement for daily estrus detection and AI over 60 to 90 days or more. Therefore, with the availability of artificial insemination, further control of estrus and breeding management was of greater interest and value, especially to the beef producer. Additional publications, not addressed herein, provide reviews of the development of cattle estrus and breeding management (Wiltbank, 1970; Wiltbank, 1974; Odde, 1990; Mapletoft et al., 2003; Patterson et al., 2003a; Kojima, 2003; Kesler, 2003; Patterson et al., 2003b; Chenault et al., 2003; Stevenson et al., 2003; Lamb et al., 2003). Early research on the estrous cycle Research to understand estrus and estrous cycles was initiated in the United States by Dr. Fred F. McKenzie and his graduate students at the University of Missouri in the 1920s using sheep. McKenzie (1983) began leading an animal research laboratory at the University of Missouri in 1923.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 6,284,263 B1 Place (45) Date of Patent: Sep
    USOO6284263B1 (12) United States Patent (10) Patent No.: US 6,284,263 B1 Place (45) Date of Patent: Sep. 4, 2001 (54) BUCCAL DRUG ADMINISTRATION IN THE 4,755,386 7/1988 Hsiao et al. TREATMENT OF FEMALE SEXUAL 4,764,378 8/1988 Keith et al.. DYSFUNCTION 4,877,774 10/1989 Pitha et al.. 5,135,752 8/1992 Snipes. 5,190,967 3/1993 Riley. (76) Inventor: Virgil A. Place, P.O. Box 44555-10 5,346,701 9/1994 Heiber et al. Ala Kahua, Kawaihae, HI (US) 96743 5,516,523 5/1996 Heiber et al. 5,543,154 8/1996 Rork et al. ........................ 424/133.1 (*) Notice: Subject to any disclaimer, the term of this 5,639,743 6/1997 Kaswan et al. patent is extended or adjusted under 35 6,180,682 1/2001 Place. U.S.C. 154(b) by 0 days. * cited by examiner (21) Appl. No.: 09/626,772 Primary Examiner Thurman K. Page ASSistant Examiner-Rachel M. Bennett (22) Filed: Jul. 27, 2000 (74) Attorney, Agent, or Firm-Dianne E. Reed; Reed & Related U.S. Application Data ASSciates (62) Division of application No. 09/237,713, filed on Jan. 26, (57) ABSTRACT 1999, now Pat. No. 6,117,446. A buccal dosage unit is provided for administering a com (51) Int. Cl. ............................. A61F 13/02; A61 K9/20; bination of Steroidal active agents to a female individual. A61K 47/30 The novel buccal drug delivery Systems may be used in (52) U.S. Cl. .......................... 424/435; 424/434; 424/464; female hormone replacement therapy, in female 514/772.3 contraception, to treat female Sexual dysfunction, and to treat or prevent a variety of conditions and disorders which (58) Field of Search ....................................
    [Show full text]
  • The Role of Oral Contraceptives in Optic Neuritis: the Story Behind the Study, Initial Experiences, and Lessons Learned
    University of Calgary PRISM: University of Calgary's Digital Repository Graduate Studies The Vault: Electronic Theses and Dissertations 2013-07-15 The role of oral contraceptives in optic neuritis: the story behind the study, initial experiences, and lessons learned Trufyn, Jessie J. Trufyn, J. J. (2013). The role of oral contraceptives in optic neuritis: the story behind the study, initial experiences, and lessons learned (Unpublished master's thesis). University of Calgary, Calgary, AB. doi:10.11575/PRISM/28340 http://hdl.handle.net/11023/817 master thesis University of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission. Downloaded from PRISM: https://prism.ucalgary.ca UNIVERSITY OF CALGARY The role of oral contraceptives in optic neuritis: the story behind the study, initial experiences, and lessons learned by Jessie J. Trufyn A THESIS SUBMITTED TO THE FACULTY OF GRADUATE STUDIES IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE DEPARTMENT OF NEUROSCIENCE CALGARY, ALBERTA JULY, 2013 © Jessie J. Trufyn 2013 Abstract There is accumulating evidence of sex differences in multiple sclerosis, making hormones a possible research avenue for therapeutic agents. Oral contraceptives are a source of synthetic hormones, however, it is unclear whether hormone-based therapies help, hinder, or have no effect on the disease in women. In an attempt to elucidate the role of sex hormones, we are currently conducting an observational study of oral contraceptives in optic neuritis, a condition that often occurs in parallel with multiple sclerosis.
    [Show full text]
  • TGA Review of Hgps
    A REVIEW TO UPDATE AUSTRALIA’S POSITION ON THE HUMAN SAFETY OF RESIDUES OF HORMONE GROWTH PROMOTANTS (HGPs) USED IN CATTLE Prepared by Chemical Review and International Harmonisation Section Office of Chemical Safety Therapeutic Goods Administration of the Department of Health and Ageing Canberra July 2003 A draft of this report was tabled at the 25th Meeting of the Advisory Committee on Pesticides and Health (ACPH), held in Canberra on the 1st May 2003. The report was subsequently endorsed out-of-session by the ACPH. Hormone Growth Promotants TABLE OF CONTENTS ABBREVIATIONS ................................................................................................................................................................4 EXECUTIVE SUMMARY ..................................................................................................................................................7 INTRODUCTION..................................................................................................................................................................9 HEALTH CONCERNS ASSOCIATED WITH HGP S................................................................................................................9 DIFFICULTIES ASSOCIAT ED WITH ASSESSING THE SAFETY OF HGPS.........................................................................10 RISK ASSESSMENTS OF HGP S .........................................................................................................................................10 PURPOSE OF THE CURRENT
    [Show full text]
  • Downloaded from Bioscientifica.Com at 09/27/2021 06:40:16PM Via Free Access 186 R
    CONTROL OF OVULATION IN CATTLE WITH MELENGESTROL ACETATE I. EFFECT OF DOSAGE AND ROUTE OF ADMINISTRATION R. G. ZIMBELMAN and L. W. SMITH Agricultural Division, The Upjohn Company, Kalamazoo, Michigan, U.S.A. {Received 2nd June 1965) Summary. Dairy heifers with normal oestrous cycles were treated with progestagen for 15 to 18 days beginning at the 15th day of the cycle. Daily oral doses of 0\m=.\25to 8 mg of melengestrol acetate (6\g=a\-methyl-6\x=req-\ dehydro-16-methylene-17-acetoxyprogesterone: mga) inhibited oestrus and ovulation in all heifers except one receiving 0\m=.\5mg. Daily intravenous injections of 0\m=.\4mg inhibited ovulation in eight out of eight heifers. Lower doses by either route suppressed oestrus but did not uniformly inhibit ovulation. Orally, mga was about 300 to 900 times as potent as map (medroxyprogesterone acetate; Provera*) but only about ten to fifteen times as potent when both were compared by intravenous injection. Groups of eight to ten heifers were fed doses of 0\m=.\2to 2\m=.\0mg daily beginning without regard to the stage of the oestrous cycle. Of seventy\x=req-\ two heifers, sixty-nine did not ovulate during treatment. The average interval from last feeding to oestrus or ovulation ranged from 2\m=.\7days at 0\m=.\2mg to 6\m=.\3days at 2\m=.\0mg. The conception rates for various groups varied from 25 to 88 % at first service. Of the total, 42 % conceived with one service and 82 % with two services. INTRODUCTION The biological activities of melengestrol acetate (mga) in laboratory animals have recently been described.
    [Show full text]
  • MD-RES-10-2017-000550 Revised 171120
    Electronic Supplementary Material (ESI) for MedChemComm. This journal is © The Royal Society of Chemistry 2017 SUPPLEMENTARY INFORMATION Identification of non-substrate-like glycosyltransferase inhibitors from library screening: pitfalls & hits Masaki Ema [1], Yong Xu [1], Sebastian Gehrke [2] & Gerd K. Wagner [1]* [1] King’s College London, Department of Chemistry, Faculty of Natural & Mathematical Sciences, Britannia House, 7 Trinity Street, London, SE1 1DB, UK. [2] King’s College London, Institute of Pharmaceutical Science, Faculty of Life Sciences & Medicine Phone: +44 (0)20 7848 1926 e-mail: [email protected] CONTENT Table S1 Composition of inhibitor library (compounds 1-130) Fig. S1 Attempted assay minituarisation (384-well plates) Fig. S2 LgtC assay results and control experiments for false positive steroid “hits” 79 and 90 Fig. S3 Control experiments for pyrazol-3-one 113 Fig. S4 Validation of assay dilution step with CSG164/LgtC Synthesis and analytical characterisation of pyrazol-3-ones 111-130 1H and 13C NMR spectra of pyrazol-3-one 113 1 Table S1: Composition of inhibitor library Steroids Cmpd Name Cmpd Name 1 Isoflupredone acetate 46 Fluorometholone 2 Norethynodrel 47 Flumethasone 3 Prednisone 48 Medrysone 4 Fulvestrant 49 Alclometasone dipropionate 5 Lynestrenol 50 Norgestrel-(-)-D 6 Danazol 51 Fluocinonide 7 Oxandrolone 52 Clobetasol propionate 8 Triamcinolone 53 Lithocholic acid 9 Dehydrocholic acid 54 Deflazacort 10 Spironolactone 55 Ethynylestradiol 3-methylether 11 Dexamethasone acetate 56 Equilin 12 Canrenoic adic
    [Show full text]
  • Molecular Regulation and Disruption of the Progesterone Receptor Signaling Pathways During Frog Embryonic Development
    MOLECULAR REGULATION AND DISRUPTION OF THE PROGESTERONE RECEPTOR SIGNALING PATHWAYS DURING FROG EMBRYONIC DEVELOPMENT by Paisley Elizabeth Thomson A thesis submitted to the School of Environmental Studies In conformity with the requirements for the degree of Master of Environmental Studies Queen’s University Kingston, Ontario, Canada (May, 2018) Copyright © Paisley Thomson, 2018 Abstract Gestagens are a class of steroid hormones capable of binding and activating progesterone receptors. Gestagens include endogenous progestogens, such as progesterone (P4), which have critically important roles in vertebrate physiology and reproduction and synthetic P4 analogues (progestins), such as melengestrol acetate (MGA). Both gestagens are administered as growth promotants in beef cattle and have been measured in surface water receiving runoff from animal agricultural operations. This project aims to understand the roles and the regulatory mechanisms of P4 in early amphibian development and to assess the consequences of exposures to environmental gestagens on the P4-receptor signaling pathways in frog embryos. We first established the developmental transcript profiles of the three P4 receptors in Western clawed frog (Silurana tropicalis) embryos. P4-receptor mRNAs were detected but differentially expressed throughout embryogenesis. Secondly, we conducted P4 and MGA acute exposures to an environmentally realistic range of concentrations to determine the effects of embryonic exposure to gestagens on development, mortality, and gene expression of reproduction-related genes. Acute exposure to P4 induced a 2- to 5-fold change increase of steroid hormone receptor mRNA levels, whereas MGA exposure induced a dissimilar transcriptional profile than P4. Therefore, we conclude that that MGA and P4 may signal through different molecular cascades in frogs.
    [Show full text]
  • Identification and Characterization of Steroid Receptor Ligands Megan Dennis
    University of New Mexico UNM Digital Repository Biomedical Sciences ETDs Electronic Theses and Dissertations 5-1-2010 Identification and characterization of steroid receptor ligands Megan Dennis Follow this and additional works at: https://digitalrepository.unm.edu/biom_etds Recommended Citation Dennis, Megan. "Identification and characterization of steroid receptor ligands." (2010). https://digitalrepository.unm.edu/ biom_etds/11 This Dissertation is brought to you for free and open access by the Electronic Theses and Dissertations at UNM Digital Repository. It has been accepted for inclusion in Biomedical Sciences ETDs by an authorized administrator of UNM Digital Repository. For more information, please contact [email protected]. i by DISSERTATION Submitted in Partial Fulfillment of the Requirements for the Degree of The University of New Mexico Albuquerque, New Mexico ii ACKNOWLDEGEMENTS First, I would like to thank my parents, Mary and Greg. Without their guidance and support throughout the years this document would not exist. Thank you. Secondly, I would like to thank my mentor, Dr. Eric Prossnitz. Eric has always been involved to the perfect degree in my research, staying out of the way and letting me learn to be an independent researcher but also always there to give advice and help me find my way. Throughout my career, I hope to be as effective a mentor to students as Eric has been for me. Next, I would like to acknowledge my committee members, who have all been involved extensively with different parts of this work. Dr. Larry Sklar taught me a great many things about flow cytometry, presenting data and fielding questions. He also told me very early in my graduate work to keep my computer files organized and as I near the end of my time as a student, I realize that this may be the most valuable lesson of all.
    [Show full text]
  • Pharmaceuticals and Chemicals of Concern in Minnesota Lakes
    Surface water January 2021 Pharmaceuticals and chemicals of concern in Minnesota lakes A statewide 2017 study of surface water shows that several medicines and other chemical contaminants found in Minnesota’s lakes may be harmful to aquatic fish and wildlife. Authors Mark Ferrey, Summer Streets, April Andrews, Minnesota Pollution Control Agency Will Backe, Minnesota Department of Health Contributors/acknowledgements This report contains the results of a study that characterizes the presence and effects of unregulated contaminants in Minnesota’s Lakes. The study was made possible through funding by the Minnesota Clean Water Fund. The authors thank the following for assistance and advice in designing and carrying out this study: Lee Engel, Dereck Richter, Jordan Donatell, Joe Hadash, Isaac Martin, Lindsay Egge, and Josh Stock for fieldwork and sampling; Shawn Nelson for ArcGIS and figure development; SGS AXYS Analytical Services for help in developing the list of analytes for this study and the logistics to make it a success. The MPCA is reducing printing and mailing costs by using the Internet to distribute reports and information to a wider audience. Visit our website for more information. MPCA reports are printed on 100% post-consumer recycled content paper manufactured without chlorine or chlorine derivatives. Minnesota Pollution Control Agency 520 Lafayette Road North | Saint Paul, MN 55155-4194 | 651-296-6300 | 800-657-3864 | Or use your preferred relay service. | [email protected] This report is available in alternative formats
    [Show full text]