Mohamed Attia Shaaban Mahmoud ___________________________________________________ Bioactive Secondary Metabolites from Marine and Terres- trial Bacteria: Isoquinolinequinones, Bacterial Compounds with a Novel Pharmacophor O O O O O N H H C N 3 N O O O O O H N N N N H Dissertation Bioactive Secondary Metabolites from Marine and Terrestrial Bacteria: Isoquinolinequinones, Bacterial Compounds with a Novel Pharmacophor Dissertation zur Erlangung des Doktorgrades der Mathematisch-Naturwissenschaftlichen Fakultäten der Georg-August-Universität zu Göttingen vorgelegt von Mohamed Attia Shaaban Mahmoud aus El-Mansoura (Ägypten) Göttingen, 2004 D7 Referent: Prof. Dr. H. Laatsch Korreferent: Prof. Dr. H. Lackner Tag der mündlichen Prüfung: 02.1l.2004 Die vorliegende Arbeit wurde in der Zeit von April 2000 bis Oktober 2004 im Insti- tut für Organische und Biomolekulare Chemie der Georg-August-Universität zu Göt- tingen unter der Leitung von Herrn Prof. Dr. H. Laatsch angefertigt. Herrn Prof. Dr. H. Laatsch danke ich für die Möglichkeit zur Durchführung dieser Arbeit sowie die ständige Bereitschaft, auftretende Probleme zu diskutieren. Für meine Eltern, meine Frau und meine Kinder I 1. Introduction.................................................................................................. 1 1.1 Developmental history of antibiotics ................................................... 1 1.2 Searching for new Sources of Antibiotics............................................ 3 1.3 Approaches to get new natural products .............................................. 6 1.4 Antibiotic screening............................................................................. 8 1.5 New Natural Metabolites from Marine Bacteria.................................. 9 1.6 The Phylogenetic Tree of Bacteria..................................................... 14 1.7 North Sea Bacteria ............................................................................. 15 1.7.1 The marine Roseobacter Clade ...................................................... 18 2 Aim of the present investigation ................................................................ 21 3 Classification of the investigated bacterial strains..................................... 22 3.1 Working up of selected strains........................................................... 23 3.1.1 Primary screening...........................................................................23 3.2 Large scale cultivation and extraction................................................ 24 3.3 Dereplication concept.........................................................................26 4 Investigation of selected Bacterial Strains ................................................. 28 4.1 Cytophaga marinoflava sp. AM13,1.................................................. 28 4.1.1 Nβ-Acetyltryptamine ...................................................................... 29 4.1.2 2-Methylbutyramide and 3-methylbutyramide .............................. 31 4.1.3 cis-Cyclo(Isoleucyl-Valyl) ............................................................. 32 4.1.4 cis-Cyclo(Tyrosyl-Prolyl) .............................................................. 33 4.1.5 cis-Cyclo(Phenylalanyl-Prolyl)...................................................... 34 4.1.6 Indolyl-3-ethylisovaleramide (Madugin)....................................... 35 4.1.7 Trivial compounds......................................................................... 37 4.1.8 2,5-Bis(3-methenylindolyl)-pyrazine............................................. 40 4.1.9 N-Phenylethyl-isovaleramide......................................................... 44 II 4.1.10 Tryptanthrine.............................................................................. 45 4.1.11 o-Acetylaminobenzamide .......................................................... 47 4.1.12 2,2-Dimethyl-2,3-dihydro-1H-quinazolin-4-on ......................... 48 4.1.13 Pharacine.................................................................................... 49 4.1.14 p –Hydroxyphenyl acetamide .................................................... 53 4.2 Marine Streptomyces sp. B1848......................................................... 55 4.2.1 1-Acetyl-β-carboline...................................................................... 56 4.2.2 2'-Deoxyadenosine......................................................................... 59 4.2.3 6-Hydroxy isatine........................................................................... 60 4.2.4 2'-Deoxythymidine, and 2'-Deoxyuridine...................................... 62 4.2.5 4-Methylamino-7,8-dimethyl-isoquinoline-3,6-dione ................... 63 4.2.6 7-Methylamino-3H-pyrrolo[2,3-c]isoquinoline-6,9-dione ............ 65 4.2.7 p-Hydroxy benzoic acid................................................................. 69 4.3 Alteromonas distincta sp. Hel69 ........................................................ 70 4.3.1 Indole ............................................................................................. 71 4.3.2 Brevianamide F.............................................................................. 72 4.4 Terrestrial Streptomyces sp. GW3/1538 ............................................ 73 4.4.1 6-Ethyl-4-hydroxy-3,5-dimethyl-2-pyrone .................................... 75 4.4.2 2,5-Furandimethanol ...................................................................... 77 4.4.3 3-Hydroxy-2-methyl-γ-pyrone; Maltol .......................................... 78 4.4.4 cis-Cyclo(Leucyl-Prolyl)................................................................ 79 4.4.5 (S)-Dihydro-4-hydroxy-2(3H)-furanone........................................ 80 4.5 Strain Bio134..................................................................................... 81 4.5.1 13-Methyltetradecanoic acid.......................................................... 82 4.5.2 3'-Acetoxy- 2'-deoxy-thymidine .................................................... 83 4.5.3 Pyrrole-2-carboxylic acid............................................................... 87 4.6 Marine Streptomyces sp. B8876......................................................... 87 III 4.6.1 Undecylprodigiosin........................................................................ 89 4.7 Marine Streptomyces sp. B7936......................................................... 91 4.7.1 Actinomycin D............................................................................... 92 4.7.2 Actinomycin D analogue ............................................................... 94 4.7.3 FD-594 ........................................................................................... 95 4.7.4 Fungichromin (14-Hydroxyfilipin III)........................................... 99 4.8 Terrestrial Streptomyces sp. GW2/577 ............................................ 102 4.8.1 N-(2-Phenylethyl)-propionamide................................................. 103 4.8.2 p-Hydroxyphenethyl acetamide; N-Acetyl-tyramine................... 104 4.8.3 p-Hydroxyphenethyl propionamide ............................................. 105 4.8.4 3-Hydroxy-N-phenethyl-butyramide ........................................... 107 4.8.5 Crotonic acid ß-phenylethyl amide .............................................. 109 4.8.6 5-Methyl-1H-quinazoline-2,4-dione ............................................ 111 4.8.7 8-Methyl-1H-quinazoline-2,4-dione ............................................ 112 4.8.8 1,8-Dimethyl-7H-quinazolino[3,2-a]quinazoline-5,12-dione...... 114 4.9 Strain Bio215................................................................................... 117 4.9.1 Indolyl-3-acetic acid methyl ester................................................ 118 4.9.2 cis-Cyclo(Prolyl-Valyl)................................................................ 119 4.9.3 Flazin............................................................................................ 119 4.9.4 1-(9H-β-Carbolin-1-yl)-3-hydroxy-propan-1-one ....................... 123 4.9.5 p-Hydroxyphenyl acetic acid and Adenine; Angustmycin B....... 125 4.10 Strain Pic009.................................................................................... 126 4.10.1 o-Hydroxyphenyl acetic acid ................................................... 127 4.10.2 Isoxanthohumol........................................................................ 127 4.10.3 Uridine ..................................................................................... 131 4.11 Terrestrial Streptomyces sp. GW10/580 .......................................... 132 4.11.1 Phenazine-1- carboxylic acid; Tubermycin B.......................... 133 IV 4.11.2 Surfactin C............................................................................... 134 4.11.3 Pyridine-3-carboxylic acid; Nicotinic acid .............................. 139 4.11.4 2-Acetamidophenol; 2-Hydroxyacetanilide............................. 139 4.11.5 Indolyl-3-carbaldehyde ............................................................ 140 4.11.6 Cytoxazone............................................................................... 141 4.11.7 Feigrisolide B........................................................................... 143 4.11.8 Feigrisolide A........................................................................... 145 4.11.9 Feigrisolide C........................................................................... 146 4.12 Marine Streptomyces sp. B 8335...................................................... 148 4.12.1 Actinomycin