Biologically Active Molecules: Identification, Characterization and Synthesis Proceedings of a Seminar on Chemistry of Biologically Active Compounds and Modern Analytical Methods, Interlaken, September 5–7, 1988

257 Pages·1989·14.29 MB·English

by  Wolfgang Steglich (auth.)

BIOLOGICALLY ACTIVE MOLECULES
Editor:
Prof. Dr. Urs Peter Schlunegger
Department of Organic Chemistry
University of Bern
Freiestrasse 3
CH-3012 Bern
BIOLOGICALLY
ACTIVE MOLECULES
Identification, Characterization and Synthesis
Proceedings of a Seminar
on Chemistry of Biologically Active Compounds
and Modern Analytical Methods
Interlaken, September 5-7, 1988
Sponsored and Organized by
The Association of Swiss Chemists
Springer-Verlag
Berlin Heidelberg New York
London Paris Tokyo
Prof. Dr. Urs Peter Schlunegger
Department of Organic Chemistry
University of Bern
Freiestrasse 3
CH-3012 Bern
ISBN-13: 978-3-642-74584-3 e-ISBN-13: 978-3-642-74582-9
DOl: 10.1007/978-3-642-74582-9
Library of Congress Cataloging-in-Publication Data
Seminar on Chemistry of Biologically Active Compounds and Modern Analytical Methods (1988 :
Interlaken, Switzerland) Biologically active molecules: identification, characterization, and
synthesis: proceedings of a Seminar on Chemistry on Biologically Active Compounds and
Modern Analytical Methods, Interlaken, September 5-7, 1988/ sponsored and organized by the
Association of Swiss Chemists; editor, Urs Peter Schlunegger. p. cm.
ISBN-13:978-3-642-74584-3(U.S. : alk. paper)
1. Drugs-Analysis'-Congresses. 2. Drugs-Synthesis-Congresses. I. Schlunegger, Urs P.
II. Schweizerischer Chemiker-Verband. III. Title. RS189.S398 1988 547-dc 19 89-6159
This work is subject to copyright. All rights are reserved, whether the whole or part of the
material is concerned, specifically the rights of translation, reprinting, re-use of illustrations,
recitation, broadcasting, reproduction on microfilms or in other ways, and storage in data banks.
Duplication of this publication or parts thereof is only permitted under the provisions of the
German Copyright Law of September 9,1965, in its version of June 24,1985, and a copyright fee
must always be paid. Violations fall under the prosecution act of the German Copyright Law.
© Springer-Verlag Berlin Heidelberg 1989
Softcover reprint of the hardcover 1st edition 1989
The use of registered names, trademarks, etc. in this publication does not imply, even in the
absence of a specific statement, that such names are exempt from the relevant protective laws and
regulations and therefore free for general use.
The publisher cannot assume any legal responsibility for given data, especially as far as directions
for the use and the handling of chemicals are concerned. This information can be obtained from
the instructions on safe laboratory practice and from the manufacturers of chemical and
laboratory equipment.
215113140-543210 - Printed on acid-free paper
PRE F ACE
Over the past few years there has been a remarkable and rapid
development of modern analytical methods, and the fields of
nuclear magnetic resonance and mass spectrometry have been no
exception. In addition to being able to do "more and faster", new
innovative techniques have also arisen to contribute to a growing
understanding of the relationship between chemical structure and
biological activity.
In order to explore a few of the more interesting points of those
developments and applications, a seminar "From Biological
Activity to Structure" was organized from September 5-7, 1988 at
Interlaken. The four invited speakers, Richard M. Caprioli,
Howard R. Morris, Wolfgang Steglich and Dudley H. Williams were
generous enough to attend and discuss many facets of their
research, especially methodological and technical developments
and their applications to specific problems. Participants were
introduced to continuous flow FAB (fast atom bombardment) and its
use for example, in the real time monitoring of biochemical
reactions in vitro and in vivo; the structural elucidation of
secondary metabolites from fungi; the analysis of molecule
receptor interactions; the determination of post translational
modif ications of peptides; and the location of S-S bridges in
determining the tertiary structure of proteins, amongst other
things.
All this provided an excellent insight into modern analytical
research: the strategy and tactics of solving problems.
In summary, a very diverse picture of modern bio-organic
analytical research was presented to the participants and is now
collected into a permanent record which is hoped will promote
further dissemination and development of this fascinating
research.
Bern, 22 December, 1988 Urs Peter Schlunegger
CON TEN T S
Wolfgang Steglich
Some Chemical Phenomena of Mushrooms and Toadstools .......... 1
Timm Anke and Wolfgang Steglich
~-Methoxyacrylate Antibiotics: From Biological Activity
to Synthetic Analogues •.........................•.......•.... 9
Joachim Vater
Lipopeptides, an Interesting Class of Microbial Secondary
Metabolites ..........................•.......•......•......• 27
Richard M. Caprioli
Fast Atom Bombardment: Basic Concepts and
Practical Considerations ............................•....... 39
Richard M. Caprioli
Continuous-Flow Fast Atom Bombardment Mass Spectrometry ...•. 59
Richard M. Caprioli
Coupling Chromatographic Techniques with Fast Atom
Bombardment Mass Spectrometry for the Structural
Analysis of Biological Compounds ......................••.... 79
Howard R. Morris, Anne Dell, Maria Panico, Roy McDowell
and Ashraf Chatterjee
The Application of High Mass Fast Atom Bombardment Mass
Spectrometry to Molecular Biology .............•.............. 97
VIII
Thaiya Krishnamurthy, Marguerite E. Brooks, Donald F. Hunt,
Jeffrey Shabanowitz, Shuian Chen and Terry Lee
Identification of Active-Site in a Neurotoxic Snake Venom
by Affinity Labelling and State-Of-The-Art Tandem Mass
Spectrometry Technology ..•••.•.•...••.•....•...••.••.•••••. 149
Dudley H. Williams
Peptide Defence Systems ...••..•••••.•.••••.••••••••..•••..• 161
Erich Hochuli
Genetically Designed Affinity Chromatography Using
a Novel Metal Chelate Absorbent ••••.••••.••••••••.••••••••• 217
Urs Peter Fringeli
Structure-Activity Relationship in Biomembranes Investigated
by Infrared-ATR Sprectroscopy .••......•..•••.••...••.•••..• 241
SOME CHEMICAL PHENOMENA OF
MUSHROOMS AND TOADSTOOLS
Wolfgang steglich
Institut fur Organische Chemie und Biochemie der Universitat Bonn
Gerhard Domagk-Stra~e I, D-53 Bonn I, West Germany
Abstract: Mushrooms are a rich source of new natural products which
often possess unusual structures and interesting biological
properties.
The fruit-bodies of mushrooms and toadstools exhibit a wide variety
of different colours, colour reactions, tastes, and odours which
often help the mycologist to determine the species. Several
mushrooms are rarely attacked by insects whereas others are easy
prey to their larvae. All of these phenomena are caused by chemicals
which have been studied only in recent years. In this lecture I want
to draw your attention to some cases where such phenomena led to the
isolation of interesting new compounds.
In the protection of fruit-bodies against insects, bitter or pungent
sUbstances play an important role. Thus the agarics cortinarius
crystallinus, c. croceo-coeruleus and c. vibratilis produce the
strongly bitter crystallopicrin 4 (1), a triterpenoid which is
concentrated in the slimy outer layer of the cap. Its structure has
been determined by 2D-INADEQUATE experiments and resembles that of
the iridals from Iris species (2). The carbon skeleton of 4 can be
HO
2
J I
{OH
~
~ HOOH
::P
HO
HO
3 HO 4
U.P. Schlunegger (Ed.)
Biologically Active Molecules
© Springer-Verlag Berlin Heidelberg 1989
2
derived from squalene epoxide 1 by a shortened version of the
lanosterol biosynthesis. cyclization of 1 leads to a carbenium ion 2
which then undergoes a backbone rearrangement terminating in a
fragmentation to aldehyde 3. Reduction of 3 and consecutive
hydroxylatiQns at the allylic positions and the terminal double bond
then leads to 4. Bitter terpenoids are also found in sporophores of
Hypholoma tasciculare(3) and Gymnopilus spectabilis(4) whereas
Cortinarius infractus(5) and Clavariadelphus pistillaris(6) owe
their bitter taste to indole alkaloids and a spermidine derivative,
respectively.
Pungent cOlllpounds are even more potent antifeedants. Well known
examples are the sharp tasting Lactarius and Russula species which
have developed an interesting chemical defence system(7). The white
milk of Lactarius vellereus contains the mild tasting sesquiter
penoid stea~lvelutinal 5 (8) which is apparently devoid of any
major biological activity. On damage to the fruit-body an enzyme is
released which hydrolyses the ester function of 5 setting the stage
>C1:1 t'.: L ><fl
CHO
CHO
R ~ CHO ~ CH20H
x:fJ:~CR
enzyme 6 enzyme 7
+ +
sec mIn
xtt xtt
5 HO , CHO
R HO R .-.: CH20H
R = (CH2)'6 CH3 8 9
for a rearrangement into the dialdehydes isovelleral 6 and velleral
8. Both compounds possess strong antibiotic, cytotoxic and mutagenic
activities (9) and are efficient antifeedants which even repel the
American opossum, a notorious mycophagist(lO). It has been found
that the toadstool detoxifies its own defence substances by
enzymatic reduction into the corresponding alcohols 7 and 9, which
are of mild taste (7). It is interesting to note that compounds
containing ene-dial moieties are often used as defence substances in
the animal and plant kingdoms(ll).
A completely different class of compounds is responsible for the
peppery taste of the small bolete Chalciporus piperatus. It contains
the pungent 2H-azepines chalciporone 10 and chalciporyl propionate
11 in admixture with the mild tasting 3H-azepines isochalciporone 12
3
and dehydroisochalciporone 13 (12). All of these substances exhibit
antibiotic activities against fungi and bacteria (13) • 2H-Azepines
are a little-known class of compounds which has not been fOllnd in
nature before.
~
10
~ ~
12 13
The defence system of Lactarius indicates that toadstools are able
to store precursors of aggressive compounds which may be released
enzymatically when needed for the protection of the fruit-bodies.
Such a system is apparently also present in Paxillus atrotomentosus,
a toadstool which is rarely infested by insects. Its fruit-bodies
contain the triester leucomentin-3 14 which on mild hydrolysis
releases its epoxysorbic acid residues yielding a mixture of
(+)-osmundalactone 15 and butenolide 16 (14). (-)-Osmundalactone is
known as an antifeedant from leaves of the Japanese fern Osmunda
japonica(15) •
15 16
Another kind of chemical defence has been invented by Agaricus
xanthoderma. This toadstool develops an intense yellow colour on
bruising and is easily recognised by its carbolic smell. The me
thanolic extract of the fruit-bodies contains a light sensitive
antibiotic which exhibits strong activity against fungi and bac
teria(16). In a first investigation of Agaricus xanthoderma, phenol,
hydroquinone, 4,4'-dihydroxydiphenyl and the pale yellow
4,4-dihydroxyazobenzene were isolated from its methanolic ex-
4
tract (17) • Later it was shown that the colour reaction of the
fruit-bodies is caused by two closely related chromogens, leu
coagaricone 17 and xanthodermin 19 (1S). On oxidation 17 is con
verted into the yellow pigment agaricone 18 whereas 19 yields the
acyl diazene derivative 20 which gives rise to a yellow anion on
..
[ox]
~N-~-o-OH
17 18
[ox] ...
19 20
dissociation. Finally, it was found that methanolic solutions of
Agaricus xanthoderma contain 4-diazo-2,5-cyclohexa-dien-l-one 21
which is easily recognised by the development of a red colour on
21 22
addition of p-naphthol owing to the formation of an azo dye. 21 and
the diazosulphonate 22 exhibit strong antibiotic and antitumor
activities(1S,19). It seems possible that xanthodermin 19 acts as a
precursor for 21 in the toadstool. The latter is then transformed
into phenol and the aromatic compounds mentioned earlier.
Agarics have a high tendency for the formation of N-N bonds. A well
known example is agaritin 23 (20) which occurs as a constituent of
the cultured mushroom Agaricus bisporus and related species.
Biosynthetically related are the aldehyde 24 and the forma zane
derivative 25 which have been isolated from A. silvicola(21).