Phytochemisrrp, Vol. 30, No. 12. pp. 3859 -3860. 1991
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Printed m Great Britam.
5. SERENDIPITY AS A BASIS FOR PHYTOCHEMISTRY
LESLIE
Rothamsted Experimental
Serendipity is a term first used by Horace Walpole in
1754 to describe the faculty of making fortuitous and
unexpected discoveries. Its etymology stems from Seren-
dib (also spelled Serendip), the Arabic name for the island
of Sir Lanka (Ceylon). Walpole was inspired by a fairy
tale ‘The Three Princes of Serendip’, whose heroes often
made discoveries by accident and sagacity of things
unsought-and so he coined serendipity. Many famous
scientists have paraphrased this definition, often stressing
the necessary association of sagacity, i.e. an acuteness of
mind. or a keenness and soundness of judgement, with
chance. Thus Pasteur wrote ‘In the fields of observation,
chance favours only the prepared mind’, and Joseph
Henry commented ‘The seeds of great discoveries are
constantly floating around us, but they only take root in
minds well prepared to receive them’.
The most frequently cited, folkloric examples of seren-
dipity are Archimedes’ accidental discovery of how to
measure the volume of an irregular shaped object whilst
in his bath and Newton’s recognition of the concept of
gravity by observing a falling apple whilst resting under a
tree. Phytochemistry, especially pharamacognosy, owes
an abundant debt to folkloric evidence. The use of
quinine as an anti-malarial is reputed to have its origin in
an accidental discovery of a South American Indian who,
thirsty and febrile, drank from a pond into which a
Cinchona tree had fallen and was thereby miraculously
cured. Perkin, intrigued to synthesize quinine even before
Kekule had proposed the closed-ring structure for
benzene, had taken aniline or toluidines as starting
materials and used the additive synthetic method then
popular. Almost inevitably, the product was not quinine,
but a black sludge. Yet fortune was with Perkin first when
he extracted the unpromising mess with alcohol and
obtained a purple pigment capable of dyeing cloth per-
manently, and later when he became a millionaire follow-
ing industrial exploitation of this and other chemical
discoveries!
An Editorial Board colleague, Trevor Goodwin, likens
serendipity to the luck of being in the right place, in the
right mind, at the right time. Certainly, that was the
background to my own incursion into the field of non-
protein amino acids of plants. Gaining a Ph.D in organic
reaction mechanisms, I decided to change my field of
chemistry and joined the Human Nutrition Unit of the
Medical Research Council to undertake a study of the
amino acid composition of seed protein of peanuts: this
was during the period of Britain’s involvement with the
Groundnut Scheme in Tanganyika, and a little after the
first publication of the paper chromatographic method
for separating amino acids. Analysis of the peanut protein
using a quantitative version of this technique was ac-
complished without undue difficulty. There the story
could have ended, but inquisitiveness prompted a study
3859
0031 9422,91 $3.00+0.
(i:! 1991 Pergamon Press plc
FOWDEN
Station, Harpenden, U.K.
of how the seed protein behaved during the early stages of
germination and seedling growth. PC’s of free amino
acids present in seedling extracts revealed the standard
range, but wholly unexpectedly two major spots reacting
yellow-brown, not purple, with ninhydrin were ‘un-
known’. The unknowns were characterised as y(4)-
methyleneglutamic acid (1) and y(4)-methyleneglutamine
(2), the first examples of aliphatic amino acids with
unsaturated carbon chains. They joined ornithine, citrul-
line and 7-aminobutyric acid as early examples of non-
protein amino acids but, unlike these compounds, did not
behave as intermediate metabolites in protein amino acid
synthesis or degradation: they were the forerunners of a
class of plant secondary products now numbering hun-
dreds.
Working independently, F. C. Steward and co-workers
published slightly later their own characterization of 1
and 2 from tulips. These novel compounds then were
produced by plants from widely disparate plant families,
i.e. Leguminosae and Liliaceae, and interest was aroused
concerning the extent to which these, and similar com-
pounds, were distributed throughout the plant kingdom.
Three lines of approach governed the choice of plant
material for future investigations: (1) random selection,
often from within national or regional floras; (2) sys-
tematic analysis of plant groups, with underlying chemo-
taxonomic interests; and (3) search for causative agents of
plant toxicities that may be non-protein amino acids.
Examples ofwhere a search for toxic principles resulted
in the characterization of new amino acids include /I$
(methylenecyclopropyI)alanine (hypoglycin A) from
B/ighia sapida (akee), mimosine from Leucuena leucoce-
phaln, indospicine from Indiyofira spicota, and /I-oxalyl-
diaminopropionic acid from Lathyrus spp.
HO,CQCH,CH(NH,)CO,H HzNCTH,CH(NH,)CO,H
CHI CH,
1 2
CO,H
OI 5,
# H
3 4
H&C H
.-’
b
H
H
5 6
3R60 Thirty years of PhJ~tochemisfrJ

Following Steward’s work with tulips, I investigated pyl)glycine, whilst Blighia sapida produces the isomeric
other members of the Liliaceae and identified azetidine-2- [runs-L-a-(carboxycyclopropyl)glycine. Recently these
carboxylic acid (3) as a new plant imino acid in Concal- compounds have been augmented by the synthesis of all
laria mujalis. The same compound was isolated almost eight possible isomers of z-(CCP)G by Japanese workers;
simultaneously by Virtanen and colleagues from another the products representing a complete range of conforma-
liliaceous species, Pol_v(gonucum ojjicinale, but an inaccur- tionally-restricted analogues of glutamate have proved
ate molecular weight determination led those workers to valuable in elucidating more fully the excitatory function-
propose an incorrect III-atom ring structure. (In the mid- ing of glutamate receptor neurones in newborn rats.
1950s. MS and NMR methods were not available for Another Editorial Board member, Arthur Bell, is re-
structural assignments. and chemists relied upon ele- spected for his contributions to chemotaxonomy of the
mental analysis to provide an empirical formula, molecu- legumes, in which he has used the non-protein amino
lar weight determination. and synthesis for structural acids extensively as indices. Always alert for new legume
characterizations.) More than a decade later, when opin- opportunities, he stumbled upon seed of Cascanosperum
ion favoured the idea that 3 was a reliable chemical index ausrrale (the Moreton Bay Chestnut) whilst in Queens-
for liliaceous plants, a survey of some Hong Kong plants land, and, learning of folkloric accounts of their toxicity,
revealed that 1-3 were all present in the legume. D&nix decided to bring seeds back to the U.K. for examination.
regicr. Ideas concerning the distribution of non-protein Initial results suggested the presence of an unidentified
amino acids (and other secondary plant products?) re- basic amino acid (reacting brownish-yellow with ninhy-
ceived a further jolt when 3 was isolated in kg amounts drin on PC’s), which was separated routinely on a cation-
during the fractionation of the ‘noxious nitrogen’ com- exchange resin column. However, subsequent structural
ponents from IO” tonnes sugar beet in a single batch tindings proved unexpected: the isolated material (castan-
process on a gigantic cation-exchange column. The most ospermine. 6) was not an amino acid but a tetrahydroxy-
sensitive techniques available for laboratory scale work octohydroindolizine alkaloid. As such, it forms one of a
have not yet revealed 3 as a constituent of sugar beet. So group of natural products now known to inhibit x-and /I-
the intriguing question is raised do all plants have the D-glucosidases in vitro and to affect the processing of
intrinsic biological machinery (genes and enzymes) rc- glycoproteins in CL‘O: hence the considerable research
quircd to synthcsire 3, albeit in the majority of species the interest in 6 as a potential antiviral agent in relation to
genes are barely expressed’? AIDS. However, the current view is that castanospermine
All three of my criteria governing plant selection is too generally intrusive on metabolism and, therefore.
converged in work with plants from the related families an unlikely candidate itself for pharmaceutical devclop-
Sapindaceae, Hippocastanaceae and Aceraceae. Hassall ment---but it could still prove to be a useful ‘lead
and co-workers had identified hypoglycin A as the toxic structure’ in the further search for drugs IO control the
principle of akee, and thus raised interest in the Sapinda- AIDS virus.
ceae as a possible source of novel amino acids. Work in This account has been reflective over 40, rather than 30
my laboratory subsequently idcntifed hypoglycin A in years of plant chemistry. Editorial Board members men-
Billia hippocasranum (Hippocastanaceae) and in Acer tioned here by name are, like myself, now retired from
pseudoplatanus (Aceraceae), and the lower homologue of their mainstream scientific appointments, and so are
hypoglycin A, namely r-(methylenecyclopropyl)glycine, more likely, or perhaps more willing, to recognize that
in another sapindaceous species, Licchi chinensis. The first serendipity has played a part in their discoveries. (I have
examples of naturally occurring acetylenic amino acids found it less easy to persuade younger colleagues to
were found in Euphoria longan, a close relative of L. accept a similar view.) Perhaps we all should refect that
chine&s, both of which arc native to Hong Kong. our subject, Phytochemistry, is largely a study of ‘living’
Extending our surveys to the genus Aesculus (Hippocas- carbon compounds and that organic chemistry was truly
tanaceae), cyclopropyl amino acids again were commonly launched by Wohler’s accidental discovery that urea
found and exhibited further structural diversity. c&3,4- could be synthesized from inorganic materials without
Methanoproline (4), isolated from A. patxiflora, has been the incursion of ‘living force’.
of particular interest. Like a number of other non-protein
imino acids, it acts as a strong antagonist (analogue) of The author acknowledges a debt to Professor Royston
prolinc and has been used to investigate how various Roberts for much general information on fortuitous discoveries
facets of proline metabolism are regulated. Compound 4 contained in his book ‘Serendipity: Accidental Discoveries in
was also the Initial choice of Shell Chemicals for a Science’. References to Investigations on non-profein amino
prospective chemical hybridizing agent (CHA) for wheat. acids are not given here, but fuller accounts of the work. together
Although cffcctive in inducing hybridization (by inhibi- with original literature citations can be found tn the revtew
ting self-pollination in wheat), percentage hybrid seed set articles listed below.
was marginally too low. Ultimately, azetidine-3-
carboxylic acid (5) was synthesized as an alternative, Fowden. L. (iYS8) BIO/.Ret:. 33, 393.
potential CHA: although the results of worldwide field Fowden. L. (1964) Ann. Ret:. Biochem. 33, 173.
trials of 5 were very promising, the compound eventually Fowden, L.. Lewis. D. and Trtsfram, H. ( 1967) Ad. En-_,vmol. 29.
was withdrawn at a late stage of devclopmcnt. 89.
Aesculus parc$va is a source of another novel cyclo- Fowden. L., Lea. P. J. and Rell. E. A. (1979) Ah. Enzymol. SO.
propyl compound, namely cis-t.-r-(carboxycyclopro- 117