ETHNOBIOLOGICAL AND CHEMICAL INVESTIGATIONS OF SELECTED AMAZONIAN PLANTS ...

255 Pages·2010·14.17 MB·English

ETHNOBIOLOGICAL AND CHEMICAL INVESTIGATIONS
OF SELECTED AMAZONIAN PLANTS
by
WILLIAM DONALD MACRAE
B.Sc, The University of Victoria, 1974
M.Sc, The University of British Columbia, 1978
A THESIS SUBMITTED IN PARTIAL FULFILMENT OF
THE REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
in
THE FACULTY OF GRADUATE STUDIES '
Biology Department
We accept this thesis as conforming
to the required standard
THE UNIVERSITY OF BRITISH COLUMBIA
March 1984
© William Donald MacRae, 1984
In presenting this thesis in partial fulfilment of the
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of B r i t i sh Columbia, I agree that the Library shall make
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The University of B r i t i sh Columbia
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Abstract
The ethnobotanical l i t e r a t u re of Amazonian South America
has been surveyed, compiled and organized. This information
allowed the i d e n t i f i c a t i on of c e r t a in taxonomic groups and
plants with specific uses which seem promising for further
research.
The use of Just ic ia p e c t o r a l is as an additive to
hallucinogenic V i r o la based snuffs has been investigated. No
a l k a l o i d al compounds could be detected in the plant and the
pharmacological e f f e c ts of an extract on mice were not
indicative of the presence of strongly psychoactive
constituents. Nor did the J. p e c t o r a l is extracts have any
inhibitory or s y n e r g i s t ic effect upon the responses of mice to
5-methoxy-N,N-dimethyltryptamine, the psychoactive constituent
of Vi rola bark. Extracts of the plant caused the relaxation of
smooth muscle and this a c t i v i ty was shown to result from the
presence of coumarin and umbelliferone. Betaine, which was
also present, was observed to e l i c it smooth muscle contraction
at high concentrations. Strong inhibitory a c t i v i ty of the
extract towards four dermatophytic fungi was observed and may
explain the use of the plant in the treatment of c e r t a in
infections. Coumarin was shown to be wholly responsible for
the aroma. The p o s s i b i l i ty that the hypnotic e f f e c ts of
coumarin may play a role in i ts use as a snuff constituent is
considered.
The use of Vi rola elongata as both an hallucinogenic
snuff and an arrow poison was examined. Extracts of the bark
were evaluated for their effects on mouse behavior. A non-
alkaloidal fraction caused a reduction in spontaneous motor
activity in mice while the alkaloidal fraction from the same
amount of plant material had no significant effect. The non-
alkaloidal fraction was examined and some of its biological
activity was attributed to the presence of polyphenolic
compounds. Eleven compounds were isolated from the non-polar
part of this extract. In addition to j3-sitosterol, two
isomeric stilbenes, two neolignans, four bis-tetrahydrofuran
lignans and two tetrahydrofuran lignans, were identified. The
bis-tetrahydrofuran lignans were shown to reduce spontaneous
motor activity and isolation induced aggression in mice. The
possibility that they are at least partly responsible for the
use of Virola elongata as an arrow poison is' raised.
Thirty-four species of Amazonian Euphorbiaceae were
screened for inhibitory activity towards Escherichia coli,
Staphylococcus aureus , two yeasts, four dermatophytic fungi,
two animal viruses, tumour formation in potato discs and
toxicity to brine shrimp. A large proportion of the extracts
were active against S. aureus, the dermatophytes, at least one
of the viruses, the potato tumours and the brine shrimp. The
biological activities observed are discussed with respect to
the use of certain species in Amazonian ethnomedicine.
The antiviral activity of one of the plants screened, a
species of Amanoa was examined. The inhibitory activity
towards infection by murine cytomegalovirus was found to
i v
result from the presence of a single compound, identified as
the lignan, a- ( - )-peltatin. At low doses(lO ng/ml for two
hours), this compound prevented the replication of viruses in
already infected cells. I t 's activity was observed to be very
similar to that of another naturally occurring lignan,
podophyllotoxin. Lignans of a variety of structural types were
examined for antiviral activity but anti-murine
cytomegalovirus activity of compounds other than the
podophyllotoxin type was not observed.
V
Table of Contents
Abstract i i
List of Tables x
List of Figures xii
Acknowledgement xiv
CHAPTER I
GENERAL INTRODUCTION 1
1. ETHNOBIOLOGY; AN INTERDISCIPLINARY APPROACH 1
2. OBJECTIVES OF ETHNOBIOLOGY 6
3. IMPORTANCE OF ETHNOBIOLOGY IN THE TROPICS 8
4. APPROACH OF THIS STUDY 11
LITERATURE CITED 25
CHAPTER II
Justicia pectoralis : A STUDY OF THE BASIS FOR ITS
USE AS A Virola SNUFF ADMIXTURE 30
1. INTRODUCTION 30
2. MATERIALS AND METHODS .31
a. Plant material 31
b. Chromatography and spectroscopy 32
c. Behavior experiments 33
d. Rat stomach strip experiments 33
e. Antimicrobial tests 34
f. Antiviral tests 35
3. RESULTS 35
vi
a. Examination of Justicia pectoralis for alkaloids
35
b. Compound 1 36
c. Behavioral effects of Just ic ia pectorali s 37
d. Effect of Justic ia pectoralis extracts on
5-MeODMT induced behavioral responses 39
e. Effect of 5-MeODMT on mouse activity 41
f. Gross behavioral effects of 5-MeODMT 44
g. Effect of co-injections of 5-MeODMT and
Justicia pectoralis extracts 44
h. Effect of Justic ia pectorali s extracts
on smooth muscle 47
i. Analysis of aromatic constituents 51
j. Quantification of coumarins of J. pectoralis ....53
k. Examination of J. pectoralis for lignans 55
1. Screening for other biological activities 57
4. DISCUSSION 59
LITERATURE CITED 63
CHAPTER III
AN ETHNOPHARMACOLOGICAL EXAMINATION OF Virola elongata
BARK, A SOUTH AMERICAN ARROW POISON 69
INTRODUCTION 69
PART A. ISOLATION AND IDENTIFICATION OF THE MAJOR NON-
POLAR CONSTITUENTS OF Virola elongata BARK 73
1. INTRODUCTION 7 3
2. EXPERIMENTAL 73
v ii
a. Extraction of plant material 73
b. Chromatography 74
3. COMPOUNDS ISOLATED 7 4
4. RESULTS 80
a. 3,4',5-trimethoxy-cis-stilbene and 3,4',5-
trimethoxy-trans-stilbene 83
b. Eusiderin and virolongin 87
c. Epi-sesartemin, sesartemin, epi-yangambin
and yangambin 88
d. Dihydrosesartemin and 0-yangambin 91
PART B. EXAMINATION OF THE BIOLOGICAL ACTIVITY OF
Virola elongata BARK EXTRACTS 100
1. INTRODUCTION 100
2. MATERIALS AND METHODS 100
a. Preparation of extracts 100
b. Fractionation of extracts 101
c. Chromatographic analysis of extracts 102
d. Assay of spontaneous motor activity 103
e. Assay of anti-aggressive activity 104
3. RESULTS 105
a. Examination of the aqueous fraction for
toxicity 106
b. Examination of the diethyl ether extract for
depression of spontaneous motor activity 109
c. Quantification of major constituents of
diethyl ether extract 114
d. Effect of bis-tetrahydrofuran lignans on
v i ii
isolation induced aggression 116
4. DISCUSSION 119
LITERATURE CITED 127
CHAPTER IV
STUDIES ON THE PHARMACOLOGICAL ACTIVITY OF AMAZONIAN
EUPHORBIACEAE 134
PART A. MULTIPLE SCREENING OF AMAZONIAN EUPHORBIACEAE
FOR BIOLOGICAL ACTIVITIES 134
1 . INTRODUCTION 134
2. MATERIALS AND METHODS 139
a. Plant material 139
b. Preparation of plant extracts 140
c. Antimicrobial screening 140
d. Antiviral activity 141
e. Potato disc tumour assay 143
f. Toxicity to brine shrimp 144
g. Analysis of data 144
3. RESULTS
a. Antimicrobial activity 145
b. Antiviral activity 150
c. Inhibition of potato tumour formation 153
d. Toxicity to brine shrimp 156
e. Correlation between biological assays 156
4. DISCUSSION 167
a. Antimicrobial activity 167
ix
b. Antiviral activity 170
c. Antitumour activity 172
d. Toxicity to brine shrimp 172
5. CONCLUSION 173
PART B. a-( - )- PELTATIN, THE ANTIVIRAL
CONSTITUENT OF Amanoa sp 176
1 . INTRODUCTION 176
2. MATERIALS AND METHODS 176
a. Plant material 176
b. Antiviral assays 177
c. Chromatography 177
3. RESULTS AND DISCUSSION 178
PART C. THE ANTIVIRAL ACTION OF LIGNANS 183
1 . INTRODUCTION 183
2. MATERIALS AND METHODS ...184
a. Chemicals 184
b. Cells and viruses 184
c. Antiviral screening of lignans 186
d. Effect of time of treatment 187
3. RESULTS 188
4. DISCUSSION 193
LITERATURE CITED 201
APPENDIX A - LIST OF AMAZONIAN.ANGIOSPERMS
OF ETHNOBOTANICAL INTEREST 207
BIBLIOGRAPHY: APPENDIX A 231
APPENDIX B - SP4100 COMPUTER PROGRAM(BASIC) 236