Humaira Naureen ___________________________________________________ Dehydrorabelomycin-1-O--L-rhamnopyranoside, Actinofu- ranone C and Further New Bioactive Secondary Metabolites from Terrestrial Streptomyces spp. OH O O CH CH OH H N 3 3 O CH 2 OH O 3 O H3C OH OH O OH CH3 HO O OH OH OH O CH3 CH3 CH3 CH3 OH O HO O HO HO Dissertation Dehydrorabelomycin-1-O--L-rhamnopyranoside, Actinofuranone C and Further New Bioactive Secondary Metabolites from Terrestrial Streptomyces spp. Dissertation zur Erlangung des Doktorgrades der Mathematisch-Naturwissenschaftlichen Fakultäten der Georg-August-Universität zu Göttingen vorgelegt von Humaira Naureen aus Chakwal (Pakistan) Göttingen 2011 D7 Referent: Prof. Dr. H. Laatsch Korreferent: Prof. Dr. U. Diederichsen Tag der mündlichen Prüfung: 15-07-2011 Die vorliegende Arbeit wurde in der Zeit von Oktober 2007 bis Juni 2011 im Institut für Organische Chemie der Georg-August-Universität zu Göttingen 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. For my parents and my husband Contents I Table of Contents 1 Introduction .............................................................................................. 1 1.1 Importance of natural products in drug discovery ..................................... 1 1.2 Recently isolated metabolites from Streptomyces spp. .............................. 4 2 Objectives of the present research work .............................................. 16 3 General techniques ................................................................................. 18 3.1 Collection of strains ................................................................................. 18 3.2 Working up procedure of selected bacterial strains ................................. 18 3.3 Pre-screening ............................................................................................ 20 3.4 Chemical screening .................................................................................. 20 3.5 Biological and pharmacological screening .............................................. 21 3.6 Cultivation and scale up ........................................................................... 22 3.7 Isolation methods ..................................................................................... 23 3.8 Dereplication ............................................................................................ 23 4 Investigation of the selected bacterial strains ...................................... 25 4.1 Terrestrial Streptomyces sp. GW54/453 .................................................. 25 4.1.1 (2E,6E)-5-Hydroxy-4,6-dimethyl-octa-2,6-dienoic acid ......................... 26 4.1.2 Actinofuranone C ..................................................................................... 31 4.1.3 4-Acetyl-1,3-dihydro-imidazo[4,5-b]pyridin-2-one ................................ 36 4.1.4 2,5-Furan-dimethanol ............................................................................... 38 4.2 Terrestrial Streptomyces sp. Ank 245 ...................................................... 38 4.2.1 1,6-Dihydroxy-2-methyl-heptan-4-one .................................................... 40 4.2.2 4-Hydroxy-1-(2-methyl-oxiranyl)-pentan-2-one ..................................... 42 4.2.3 2-(2-Hydroxy-propyl-4-methyl-furan-3-carboxylic acid amide .............. 44 4.2.4 4-Vinylphenol .......................................................................................... 50 4.2.5 2-Methoxy-4-vinylphenol ........................................................................ 51 4.3 Terrestrial Streptomyces sp. Ank 250 ...................................................... 52 4.3.1 2-(5-Chloro-2-hydroxy-phenyl)-benzoxazole-4-carboxylic acid methyl ester .........................................................................................................53 4.3.2 MC-033 .................................................................................................... 57 4.4 Terrestrial Streptomyces Strain GW 51/426 ............................................ 62 4.4.1 Dehydrorabelomycin-1-O--L-rhamnopyranoside ................................. 63 4.4.2 Rhodonocardin ......................................................................................... 68 4.4.3 Cyclo(Ala-Pro) ......................................................................................... 73 II Contents 4.5 Terrestrial Streptomyces sp. WO 463 ...................................................... 75 4.5.1 Oligomycin A ........................................................................................... 76 4.5.2 Oligomycin F ............................................................................................ 79 4.5.3 5'-Methoxy-6-(N,N-dimethyl)-adenosine ................................................ 83 4.5.4 4-α-L-Glucosyl--pyranone ..................................................................... 87 4.5.5 Other metabolites ..................................................................................... 90 4.6 Terrestrial Streptomyces sp. Ank 351 ...................................................... 91 4.6.1 Geldanamycin ........................................................................................... 92 4.6.2 17-O-Demethylgeldanamycin .................................................................. 96 4.6.3 Nigericin ................................................................................................... 99 4.6.4 11, 11'-O-Dimethylelaiophylin .............................................................. 102 4.6.5 11-O-Monomethylelaiophylin ................................................................ 107 4.6.6 Nocardamine .......................................................................................... 109 4.7 Terrestrial Streptomyces sp. Ank 202 .................................................... 110 4.7.1 Ravidomycin .......................................................................................... 111 4.7.2 N-(2-Methoxyphenyl)-acetamide ........................................................... 116 4.7.3 Phenazine-1-carboxylic acid .................................................................. 117 4.7.4 1-Carboxymethyl-phenazine .................................................................. 118 4.7.5 N6, N6-Dimethyladenosine .................................................................... 118 4.8 Terrestrial Streptomyces sp. Ank 291 .................................................... 119 4.8.1 Ditrisarubicin A ...................................................................................... 120 4.8.2 Ditrisarubicin B ...................................................................................... 122 4.8.3 A447 D' .................................................................................................. 123 4.9 Terrestrial Streptomyces sp. GW 13/475 ............................................... 127 4.9.1 3-Methoxy-streptenol C ......................................................................... 127 4.9.2 Streptenol A and C ................................................................................. 131 4.10 Terrestrial Streptomyces sp. GW 12/459 ............................................... 133 4.10.1 Bisphenol A diglycidyl ether (BADGE) ................................................ 134 4.10.2 3-Methoxy-1-[4-[1-methyl-1-[4-(2-oxiranylmethoxy)phenyl]ethyl]- phenoxy]- 2-propanol (BADGE∙OMe) .................................................. 139 4.10.3 3-Chloro-1-[4-[1-methyl-1-[4-(2-oxiranylmethoxy)phenyl]- ethyl]phenoxy]- 2-propanol (BADGE∙HCl) .......................................... 143 4.10.4 3-[4-[1-[4-(3-Chloro-2-hydroxypropoxy)phenyl]-1- methylethyl]phenoxy]-1,2-propanediol (BADGE∙HCl∙H2O) ................ 144 4.11 Terrestrial Streptomyces sp. GW 8594 .................................................. 147 4.11.1 3-Hexenyl-1-O--D-glycopyranoside .................................................... 149 4.11.2 3-Hexenyl-1-O--2'-methoxy-glycoside................................................ 151 Contents III 4.11.3 3-Hexenyl-1-O--D-glycoside ............................................................... 153 4.11.4 2-Phenylethanol-1-O--D-glucoside...................................................... 155 4.12 Terrestrial Streptomyces sp. Ank 282 .................................................... 157 4.12.1 Benadrostin ............................................................................................ 158 4.12.2 p-Hydroxybenzoic acid methyl ester ..................................................... 159 4.12.3 Macrolactin A......................................................................................... 160 5 Summary ............................................................................................... 162 6 Materials and Methods ........................................................................ 174 6.1 General ................................................................................................... 174 6.2 Materials ................................................................................................ 174 6.3 Spray Reagents ....................................................................................... 175 6.4 Microbiological Materials ...................................................................... 175 6.5 Recipes ..................................................................................................