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