UNIVERSITI PUTRA MALAYSIA
ANTIMICROBIAL AND CYTOTOXIC COMPOUNDS OF SCORODOCARPUS BORNEENSIS (OLACACEAE)
AND GLYCOSMIS CALCICOLA (RUTACEAE)
CHRISTOPHE WIART
FSAS 2001 28
ANTIMICROBIAL AND CYTOTOXIC COMPOUNDS OF SCORODOCARPUS BORNEENSIS (OLACACEAE) AND GLYCOSMIS
CALCICOLA (RUTACEAE)
By
CHRISTOPHE WIART
Thesis Submitted in Fulfillment of the Requirement for the Degree of Doctor in Philosophy in the Faculty of Science and Environmental Studies
Universiti Putra Malaysia
March 2001
To my beloved wife Mazdida, to my beloved mother Flora, my father Patrice, my sister Vanessa, my sons Adam and Pierre, In memory of my beloved and greatly missed grand parents Renee and Jose Monllor ... In memory of my mentors Pro Loic Girre, Dr. Lucille Allorge, Dr. Kochumen ..
11
Abstract of thesis presented to the Senate ofUniversiti Putra Malaysia in fulfilment of the requirements for the degree of Doctor in Philosophy
ANTIMICROBIAL AND CYTOTOXIC COMPOUNDS OF SCORODOCARPUS BORNEENSIS (OLACACEAE) AND GLYCOSMIS
CALCICOLA (RUTACEAE)
By
CHRISTOPHE WIART
March 2001
Chairman: Professor Mawardi Rahmani, Ph.D
Faculty: Science and Environmental Studies
The extracts of plants collected from the forest of Northern Malay
Peninsula were tested against bacteria, fungi and CEM-SS leukemia cel1 line.
Gram-positive Bacillus cereus and Gram-negative Pseudomonas aeruginosa which
are commonly involved in skin infections, have been used for screening
antibacterial activity whereas Candida lipo/ytica, Saccharomyces cerevisae,
Saccharomyces lipoiytica, and Aspergillus ochraceous have been used for
screening antifungal activity.
The crude extract from the seeds of Scorodocarpus bomeensis showed a
strong cytotoxic activity against CEM-SS leukemia cell-line and antimicrobial
activities. A bioassay-guided separation had been carried out to separate the active
constituents of the seeds. This resulted in the isolation and identification of an
aliphatic sulfur compound, bis-(methylthiomethyl)disulfide, and a new indole
111
alkalOld, 13-docosenoyl serotonme from the petroleum ether extract A new
sesqUlterpene, scopotm, has been Isolated from the chloroform extract of the seeds
Another sesqUlterpene, cadalene-p-carboxyhc aCId, has been Isolated from the bark
of thIS speCIes
bls-(MethylthIOmethyl)-dIsulfide appeared as the most actIve compound
and the major constttuent of the seeds extract of Scorodocarpus borneensls It
acted slgmficantiy on a methIclllm resIstant stram of Staphylococcus aureus and
showed a strong CytOtOXIC actIvity agamst CEM-SS leukemIa and KU8 1 2F chromc
myelogenous leukemIa cell hnes 1 3-Docosenoyl serotonme showed a moderate
CytOtOXIC actIVIty agamst CEM-SS leukemIa cell ime Scopotm showed a moderate
antImIcrobIal actIVIty and a strong CytOtOXIC effect agamst CEM-SS leukemIa cell
lme
The crude petroleum ether extract and bls-(methyithtomethyl)dlsuifide
strongly mhibited the growth of pathogen fungI and was formulated m external
preparatIOn by usmg commercIal paraffin as eXCIpIent ThIS preparatIOn exhIbIted
drastIC antIfungal actIVItIes In vitro and In VIVO The acute tOXICIty of the crude
petroleum ether extract was found to be mfenor to phenobarbItal m mIce WIth
LDso at 275 mg/kg The skm Imtancy test was performed on rabbits and It showed
that thIS preparatlOn had an Irntancy level acceptable by the FDA
IV
Antimicrobial and cytotoxic activities were also performed on
methylgerambullin and desmethoxyzanthophylline which were previously isolated
from Glycosmis calcicola (Rutaceae). Methylgerambullin showed a strong
cytotoxic activity against CEM-SS leukemia, melanoma HACCl , and colon cancer
HT29 cell lines whereas desmethoxyzanthophylline has moderate activity against
Bacillus cereus, Pseudomonas aeruginosa and CEM-SS leukemia cell line.
v
Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi keperluan untunk ijazah Doktor Falsafah
ANTIMICROBIAL AND CYTOTOXIC COMPOUNDS OF SCORODOCARPUS BORNEENSIS (OLACACEAE) AND GLYCOSMIS
CALCICOLA (RUT ACEAE)
Oleh
CHRISTOPHE WIART
Mac 2001
Pengerusi: Profesor Mawardi Rahmani
Fakulti: Sa ins dan Pengajian Alam Sekitar
Ekstrak tumbuh - tumbuhan yang dikumpul dari hutan Utara Semenanjung
Malaysia telah diuji keberkesanannya terhadap bakteria, fungi dan sel leukemia
CEM-SS. Bakteria Gram-positif Bacillus cereus dan Gram-negatif Pseudomonas
aeruginosa yang biasa terlibat di dalam jangkitan kulit telah digunakan untuk
kajian aktiviti antibakteria. Manakala fungi Candida lipolydca, Saccharomyces
cerevisae, Saccharomyces /ipolytica dan Aspergillus ochraceous telah digunakan
pula untuk aktiviti antifungi.
Didalam kajian ini, ekstrak daripada bahagian buah pokok Scorodocmpus
borneensis telah mempamerkan aktiviti sitotoksik yang kuat terhadap sel
leukemia CEM-SS dan juga terhadap aktiviti antimikrobiaI. Kerja pemisahan
biocerabinaan-kawalan telah dijalankan terhadap ekstrak ini untuk mencari
sebatian yang aktif Satu sebatian sulfur alifatik, bis-(methylthiomethyl)disulfide
VI
dan satu alkaloid indol yang bam, 13-docosenoyl serotonine, telah ditemui
daripada ekstrak petroleum eter. Manakala satu sesquiterpene yang baru,
scopotin, telah dipisahkan daripada ekstrak klorofom terhadap buah. Selain
daripada itu, satu sesquiterpene asid kadalena-f3-karboksilik telah ditemui dari
bahagian kulit pokok ini.
Didapati, bis-(methylthiomethyl)disulfide muncul sebagai sebatian yang
paling aktif dan merupakan sebatian major daripada ekstrak buah Scorodocarpus
bomeensis ini. Ia bertindak kuat terutamanya terhadap Staphylococcous aureus
yang resisten kepada methicillin. Sebatian ini juga menunjukkan sitotoksik yang
kuat terhadap sel leukemia CEM-SS dan KU81 iF. Manakala sebatian 13-
docosenoyl serotonine pula menunjukkan sitotoksik terhadap sel leukemia CEM
SS. Begitu juga dengan sebatian scopotin yang menunjukkan aktiviti
antimikrobial sederhana tetapi memberi kesan sitotoksik yang kuat terhadap sel
leukemia CEM-SS.
Ekstrak mentah petroleum eter dan sebatian bis-
(methylthiomethyl)disulfide pada khususnya menghalang kuat pertumbuhan fungi
patogen dan telah diformulasikan sebagai kegunaan luar dengan menggunakan
paraffin sebagai asasnya. Penyediaan ini telah mempamerkan aktiviti antifungal
yang amat memberangsangkan dalam in vitro dan in vivo. Didapati ketoksikan
ekstrak petroleum eter ini terhadap tikus adalah lemah berbanding dengan
fenobarbital dengan LDso sebagai 275 mg/kg. Ujian rangsangan kulit terhadap
vii
amab telah dijalankan dan didapati fonnulasi ini adalah pada kadar rangsangan
yang diterima oleh FDA.
Ujian antimikrobial dan sitotoksik juga telah dijalankan ke atas sebatian
methylgerambullin dan desmethoxyzanthophylline yang mana telah dipencilkan
daripada pokok Glycosmis calcicola (Rutaceae). Sebatian methylgerambullin
menunjukkan aktiviti sitotoksik yang kuat terhadap sel leukemia CEM-SS,
melanoma HACCl dan sel kolon kanser HT29. Manakala sebatian
desmethoxyzanthophylline pula rnenunjukkan aktiviti yang sederhana terhadap
bakteria Bacillus cereus, Pseudomonas aeruginosa dan sel leukemia CEM-SS.
viii
ACKNOWLEDGMENTS
I wish to express my sincere appreciation to Professor Dr. Mawardi
Rahmani, Chairman of my Supervisory committee, for his guidance and
encouragement throughout the thesis research. Special thanks are also extended to
the other members of the committee, Associate Professor Dr. Abdul Manaf Ali
and Associate Professor Dr. Mohd. Aspollah Sukari, for their assistance.
Thanks are due to Ms Chew Yean Ling for her work on Glycosmis
calcicola.
Thanks are also due to lab technicians and graduate students for their
support.
Finally, my deepest thanks to my wife and my sons for the sacrifices,
understanding and support they have given me throughout this work.
ix
I certify that an Examination Committee met on 25th March 2001 to conduct the final examination of Christophe Wiart on his Doctor of Philosophy thesis entitled "Antimicrobial and Cytotoxic Compounds from Scorodocarpus borneensis (Olacaceae) and Glycosmis calcicola (Rutaceae)" in accordance with Universiti Pertanian Malaysia (Higher Degree) Act 1980 and Universiti Pertanian Malaysia (Highe� Degree) Regulations 1981. The Committee recommends that the candidate be awarded the relevant degree. Members of the examination committee are as follows:
Taufiq Yap Yun Hin, Ph.D. Faculty of Science and Environmental Studies Universiti Putra Malaysia (Chairman)
Mawardi Rahmani, Ph.D. Associate Professor Faculty of Science and Environmental Studies (Member)
Abdul Manaf Ali, Ph.D. Professor Faculty of Science and Environmental Studies Universiti Putra Malaysia (Member)
Aspolla Sukari, Ph.D. Professor Faculty of Science and Environmental Studies Universiti Putra Malaysia (Member)
A. Hamid A. Hadi, Ph.D. Professor Deputy Dean Faculty of Science University Malaya (Independent Examiner)
AINI IDERIS, Ph.D. ProfessorlDean of Graduate School, Universiti Putra Malaysia
Date: 2 [) MAY 2001
ix
This Thesis was submitted to the Senate of Universiti Putra Malaysia and was accepted as fulfilment of the requirements for the degree of Doctor of Philosophy.
Date: 1 4 JUN 2001
Xl
DECLARATION
I hereby declare that the thesis is based on my original work excepts for quotations and citations which have been duly acknowledged. I also declare that it has not been previously or concurrently submitted for any other degree at UPM or other institutions .
Xli
Candidate Christophe Wiart
Date: f-S � MIj too I
TABLE OF CONTENTS
Page
DEDICATION ABSTRACT
11 1ll
ABSTRAK ACKNOWLEDGMENTS APPROV AL SHEETS DECLARATION FORM LIST OF TABLES
VI IX X XlI xv XVlll XXll
LIST OF FIGURES LIST OF ABBREVIATIONS
CHAPTER
I INTRODUCTION 1
II LITERATURE REVIEW 5
ill
Plants Natural Products of Therapeutic Importance 5 Natural Products with Therapeutic Potentials 1 0
Antitumour Compounds from Plants 1 0 Antimicrobial Compounds from Plants 1 6 Plant Natural Products with Antiprotozoal Activity 22 Natural Products of the Malay Peninsula's Plants 26
Scorodocarpus bomeensis (Olacaceae) 3 7 Botanical Characteristics 3 7 Chemical Constituents of Scorodocarpus
borneensis 40 Glycosmis calcicola (Rutaceae) 48
Botanical Characteristics 48 Chemical Constituents of Glycosmis calcicola 48
EXPERIMENTAL 50 General Experimental Procedure 50
Instruments 50 Chromatography 5 1 Plant Collection, Extraction and Bioassay 52 Biological Tests 53 Toxicological Tests 55 External Preparation 56
Extraction and Identification of Compounds from Scorodocarpus borneensis 57
The Constituents of the Seeds 57 The Constituents of the Bark 62
Purification Process in View of Large Scale Production 63
xiii
Steam Distillation 63 Extraction of bis-(Methylthiomethyl)disulfide
by Silver Nitrate Precipitation 64
IV RESULTS 66 Constituents of the Seeds of Scorodocarpus borneensis 66
Isolation and Identification of bis-(Methylthiomethyl)disulfide 68
Isolation and Identification of 1 3 -Docosenoyl Serotonine 75
Isolation and Identification of Scopotin 92 Constituents of the Bark of Scorodocarpus borneensis 105
Isolation and Identification of 8-Isopropyl-5-methyl-2-naphtoic Acid 1 05
Biological Activity Results 1 17 Screening of Crude Extracts 1 1 7 Screening of Pure Compounds 125 Bioassay Guided Fractionation of the Crude
Petroleum Ether Extract of the Seeds of Scorodocarpus borneensis 1 27
Bioactive Compounds of the Crude Petroleum Ether Extract 13 0
Inhibition of Pathogen Fungi Growth 1 3 3 Antiseptic Activity 137 Acute Toxicity 142 Skin irritancy 1 47
Purification Process in View of Large Scale production 1 5 1 Steam Distillation 15 1 Extraction of bis-(Methylthiomethyl)disulfide
by S ilver Nitrate Precipitation 151
V DISCUSSION 1 52 Antifungal Potentials of the Crude Oil and
bis-(Methylthiomethyl)disulfide 152 Toxicity 154
Acute Toxicity 154 Skin Irritancy 156
VI CONCLUSION 160
BIBLIOGRAPHY 161
BIODATA 171
XIV
Table
1
2
3
4
5
6
7
8
9
LIST OF TABLES
Natural products of therapeutic importance (Evans, 1 996)
Some anti tumour compounds from plants (Dewick, 1 996)
Some clinically important antibiotics (Berdy, 1 982)
Plant constituents with anti-HIV activity (Evans, 1 996)
Some plant constituents with anti protozoal activity (Phillipson et a!., 1 993)
Estimated number of Malaysian and World species of plants (Whitmore, 1 972)
Dicotyledone orders and families of the Malay Peninsula (Whitmore, 1 972)
Distribution offlavonoids in leaves of Olacaceae (Haron and Ping, 1 997)
lH NMR spectral data of 1 3-docosenoyl serotonine, N-fatty acyl tryptamine and bufobutanoic acid
1 0 l3e NMR spectral data of 1 3 -docosenoyl serotonine, bufobutanoic
Page
6-7
1 0- 12
1 7- 1 8
20
23-24
26
29-30
40
79
acid and N-fatty acyl tryptamine 80
1 1 lH NMR and 13e NMR spectral data of 1 3 -docosenoyl serotonine 82
1 2 NMR spectral data of scopotin
13 NMR spectral data of8-isopropyl-5-methyl-2-naphtoic acid
1 4 Biological screening of plants collected in Malays ian tropical
95
1 08
primary rain forest 1 1 9- 1 2 1
1 5
1 6
1 7
Antibacterial activities of Vitex longisepaJa leaves fractions
Minimum inhibiting concentration values of Scorodocarpus borneensis crude methanol seeds extract
Antibacterial and cytotoxic activity of various parts of Scorodocarpus borneensis
xv
1 22
1 23
123
1 8 Cytotoxicity of methylgerambullin 1 25
1 9 Antimicrobial and cytotoxic activities of Desmethoxyzanthophylline 1 26
20 Antimicrobial activity of Scorodocarpus bomeensis seeds fractions 1 28
21 Minimum inhibiting concentration values of Scorodocarpus bomeensis seeds fractions 1 28
22 Cytotoxic activity of Scorodocarpus bomeensis seeds, leaves and barks methanol extracts against CEM-SS 1 3 0
23 Antimicrobial activity of the various fractions obtained from a column chromatography of the crude petroleum ether
extract of Scorodocarpus bomeensis seeds 1 3 1
24 Antimicrobial activity of bis-(methylthiomethyl)disulfide 1 3 1
2S Activity of bis-(methylthiomethyl)disulfide against methicil lin resistant Staphylococcus aureus 1 3 2
26 Antimicrobial activity of scopotin 1 3 3
27 Antifungal activity of Scorodocarpus bomeensis seeds Petroleum ether extract against pathogenic fungi 1 3 4
28 Comparative activity of crude petroleum ether extract and nystatin standard against pathogenic fungi 1 3 4
29 Activity of bis-(methylthiomethyl)disulfide against pathogenic fungi 135
30 Comparative activity of bis-(methylthiomethyl)disulfide and nystatin standard against pathogenic fungi 136
3 1 In vitro evaluation of external preparations containing various concentration of Scorodocarpus bomeensis seeds crude petroleum ether extract against Candida /ipolytica 137
32 In vitro evaluation of deeply ensemenced external preparations containing various concentrations of Scorodocarpus bomeensis seeds crude petroleum ether extract against Candida /ipo/ytica 1 3 8
xvi
33 Symptoms observed after Microsporium inoculation 140
34 Symptoms observed after daily application of Scorodocarpus bomeensis seeds extract (5 %) containing preparation 1 42
35 Behavior of the mice before injection 1 42- 1 43
36 Determination of the LDso of Scorodocarpus bomeensis seeds crude petroleum ether extract 1 43 - 144
3 7 Recapitulative table of weights and deaths 144-145
3 8 Recapitulative table of the behavioral signs observed during the LD 50 determination 1 46
39 Evaluation of skin reaction to Scorodocarpus bomeensis seeds crude petroleum ether extract 1 48
40 Some of the commonest dermatophytoses (Connan et ai., 1 97 1 ) 1 53
41 Relationship between toxic signs and body organs (Namara, 1 976) 1 55
42 Classification of the various types of toxicity (Franck, 1 996) 1 56
43 Evaluation of skin reaction (Draize, 1 955) 1 57-158
XVII
LIST OF FIGURES
Figure Page
Chemical Structure of Plant Constituents Used in Therapeutic (pharmaceutical Codex, 1 979) 8
2 Some Chemical Structure of Antitumour Compounds from Plants (Dewick, 1 996) 1 3
3 Some Chemical Structure of Anti-HlV Compounds from Plants (Evans, 1 996) 2 1
4 Some Chemical Structure of Plant Constituents with Antiprotozoal Activity (Phillipson et aI., 1 993) 25
5 Map of the Malay Peninsula 27
6 Scorodocarpus bomeensis (personal Communication) 3 9
7 Some Chemical Constituents Reported from the Olacaceae Family 43
8 Chemical Constituents Reported from Scorodocarpus bomeensis 46
9
10
Sulfur Compounds of Allium cepa L. (Onion) and Allium sativum (Garlic)
Chemical Constituents Reported from Glycosmis calcicola (Chew, 1 995)
1 1 Extraction of bis-(Methylthiomethyl)disulfide by Silver Nitrate Precipitation
12
1 3
1 4
1 5
1 6
1 7
Isolation of the Three Bioactive Compounds of the Petroleum Ether Extract of the Seeds of Scorodocarpus bomeensis
Chromatogram Obtained from Fraction 1 4
IR Spectrum of bis-(Methylthiomethyl)disulfide
Mass Spectrum of bis-(Methylthiomethyl)disulfide
IH NMR Spectrum of bis-(Methylthiomethyl)disulfide
l3C NMR Spectrum of bis-(Methylthiomethyl)disulfide
xviii
47
49
67
68
71
7 1
72
73
1 8 CO SY Spectrum of bis-(Methylthiomethyl)disulfide 74
19 Partial Structure of 1 3-Docosenoyl Serotonine 77
20 N-Fatty Acyl Tryptamine 78
21 Bufobutanoic Acid 78
22 Erucic Acid 8 1
23 1 3 -Docosenoyl Serotonine 8 1
24 IR Spectrum of 1 3-Docosenoyl Serotonine 83
25 Mass Spectrum of 1 3-Docosenoyl Serotonine in Positive Ion Mode 84
26 Mass Fragmentation of 1 3-Docosenoyl Serotonine 85
27 IH NMR Spectrum of 1 3-Docosenoyl Serotonine 86
28 13C NMR Spectrum of 1 3-Docosenoyl Serotonine 87
29 DEPT Spectrum of 1 3-Docosenoyl Serotonine 88
30 CO SY Spectrum of 1 3-Docosenoyl Serotonine 89
31 HETCOR Spectrum of 1 3-Docosenoyl Serotonine 90
32 HMBC Spectrum of 1 3-Docosenoyl Serotonine 9 1
33 Scopotin 94
34 UV Spectrum of Scopotin 96
35 IR Spectrum of Scopotm 97
36 Mass Spectrum of Scopotin in Positive Ion Mode 98
37 IH NMR Spectrum of Scopotin 99
38 J3C NMR Spectrum of Scopotin 1 00
39 DEPT Spectrum of Seopotin 1 01
XIX
40 COSy Spectrum of Scopotin 102
41 HETCOR Spectrum of Scopotin 103
42 HMBC Spectrum of Scopotin 104
43 8-Isopropyl-5-methyl-2-naphtoic Acid 107
44 IR Spectrum of 8-Isopropyl-5-methyl-2-naphtoic Acid 109
45 Mass Spectrum of 8-isopropyl-5-methyl-2·naphtoic Acid in Positive Ion Mode 110
46 IH NMR Spectrum of 8-Isopropyl-5-methyl-2-naphtoic Acid 111
47 l3C NMR Spectrum of 8-Isopropyl·5-methyl-2-naphtoic Acid 112
48 DEPT Spectrum of8-Isopropyl-5-methyl-2-naphtoic Acid 113
49 COSY Spectrum of 8-Isopropyl-5-methyl-2-naphtoic Acid 114
50 HETCOR Spectrum of8-Isopropyl-5-methyl-2-naphtoic Acid 115
51 HMBC Spectrum of 8-Isopropyl-5-methyl-2-naphtoic Acid 116
52 Activity of the Chloroform Extract of Vitex longisepa/a Leaves Against Bacillus cereus: Bioautography (Left), Paper Disc Test (Right) 124
53 Methylgerambullin 126
54 Desmethoxyzanthophylline 127
55 CEM-SS Growth Inhibition by Scorodocarpus borneensis Seeds Crude Methanol Extract In Vitro. Control (Top), Cells Treated with Extract at ICso for 3 Days (Below) 129
56 Candida albicans Growth Inhibition by Scorodocarpus borneensis Seeds Crude Petroleum Ether Extract In Vitro 136
57 In Vitro Evaluation of External Preparations Containing Various Concentrations of Scorodocarpus borneensis Seeds Crude Petroleum Ether Extract Against Candida lipolytica 139
58 Effects of 15 Days Treatment ofa Wound with 5%
xx
59
60
61
Scorodocarpus bomeensis Seeds Extract-Containing Preparation: Before (Top), After (Below)
Determination of the LDso of the Crude Petroleum Ether Extract of Scorodocarpus borneensis Seeds
Test of Skin Irritancy: Rabbits Treated with Paraffin (Top), Oil (Below)
Test of Skin Irritancy: Rabbits Treated with AlCb
xxi
14 1
147
1 49
1 50
CHCb CDCb cm COSY d dd EtOH FDA g GC kg HETCOR IC50 IR LD50 I m Me MeOH mg MIC ml mm m.p. MS NCI NMR PE ppm rpm s TLC UI )..ll
)..lg )..lM
LIST OF ABBREVIATIONS
chlorofonn deuterated chloroform centimeter Correlation Spectroscopy doublet doublet of doublet ethanol Food and Drug Administration gram Gas Chromatography kilogram Heteronuclear Correlation Inhibiting Concentration 5 0% Infra Red Lethal Dose 50% liter multiplet methyl methanol milligram Minimum Inhibiting Concentration milliliter millimeter melting point Mass Spectrum National Cancer Institute Nuclear Magnetic Resonance Petroleum Ether part per million rotation per minute singlet Thin Layer Chromatography Unit International microliter mlcrogram micromolar
xxii
CHAPTER I
INTRODUCTION
WIdely recogmzed as bemg of senous and ImmedIate concern IS the cnsls
of new and re-emergmg mfectIOus diseases for which no effectIve therapies are
avaIlable and the development of reSIstance of many pathogens, such as
Staphylococcus aureus to methiCIllIn PemcIllms dIscovered m the fortIes as well
as more recent antibIOtICS, are now totally meffectlve m common bactenal
mfectIOns
Another concern IS the need to develop new cancer chemotherapeutIc
agents With activIty agamst the dIsease-types stIll resistant to current therapies and
to overcome the development of rnu1tldrug reSistance, which IS mcreasmgly
observed m the treatment of many tumors Death rates and sIde effect are stIll
hIgh and new chemotherapeutic drugs need to be dIscovered
Thus, there IS an urgent need to IdentIfy novel antImIcrobIal and CytotOXIC
molecules as leads for effective drug development Such actIve molecules may be
syntheSIzed by orgamc chemISts or extracted from bactena, fungI, alllmals, or
plants The plant kmgdom has been descnbed by Farnworth as a "vIrtually
untapped reservOIr of novel drugs awaItmg Imagmative and progreSSIve
orgamzatIOns"(Cordell, 1995) The author also estimated that five to fifteen
percent of the approxImately two hundred and fifty thousand speCIes of higher
plant have been systematically investIgated for the presence of bIOactIve
compounds (Cordell, 1 995)
BIOactlve compounds of plant ongIn have played, and contInue to play an
Invaluable role In the drug discovery process related to all disease types and, In
particular, In the area of cancer and InfectIOus diseases For example, natIOnal
prescnptlOn audit data for a penod of fifteen years between 1 959- 1 974 showed
that over 25% of US PrescnptIOns dispensed In 1 973 contaIned active Ingredients
denved from higher plants, while 1 3 3% and 2 7% were denved from microbial
and ammal sources, respectIvely (Cordell, 1 995) Among the new approved drugs
reported between 1 983 and 1 994, drugs of natural ongIn predomInate In the area
of antlbactenal (78%), whIle 6 1 % of the anticancer drugs are naturally denved or
modeled on natural product parents Of the eighty seven approved anticancer
drugs four are of plant ongIn Of the two hundred and nInety nllle anticancer drug
candidates nIne are of plant higher ongIn (Cordell, 1 995)
Higher plants which are able to produce new drugs are maInly found In
pnmary and secondary tropIcal ram forests of AfrIca, AsIa and espeCially South
and Central Amenca and Southeast Asia The bIOdiverSity there has forced plants
to protect themselves agaInst mlcroorgamsms, ammals or Insects by prodUCIng a
great vanety of chemical weapons In Southeast Asia, our MalaYSian forest IS
endowed With one of the oldest and nchest flora In the world (Ridley, 1 922) ThiS
2