Structure Elucidation of Secondary Metabolites from Rudbeckia Species by Spectroscopic Techniques and Review of Sesquiterpene Lactones

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1989 Structure Elucidation of Secondary Metabolites From Rudbeckia Species by Spectroscopic Techniques and Review of Sesquiterpene Lactones. Marta Vasquez Louisiana State University and Agricultural & Mechanical College

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Recommended Citation Vasquez, Marta, "Structure Elucidation of Secondary Metabolites From Rudbeckia Species by Spectroscopic Techniques and Review of Sesquiterpene Lactones." (1989). LSU Historical Dissertations and Theses. 4815. https://digitalcommons.lsu.edu/gradschool_disstheses/4815

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Structure elucidation of secondary metabolites fromRudbeckia species by spectroscopic techniques and review of sesquiterpene lactones

Vasquez, Marta, Ph.D.

The Louisiana State University and Agricultural and Mechanical Col., 1989

U M ’I 300 N. Zeeb Rd. Ann Arbor, MI 48106

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. STRUCTURE ELUCIDATION OF SECONDARY METABOLITES FROM RUDBECKIA SPECIES BY SPECTROSCOPIC TECHNIQUES AND REVIEW OF SESQUITERPENE LACTONES

A Dissertation

Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of

Doctor of Philosophy

The Department of Chemistry

by Marta Vasquez Universidad de Antioquia, Colombia, 1971 . , Universidad del Valle, Colombia, 1981 August 1989

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. To my family members

for believing in the worth of this

project

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ACKNOWLEDGMENTS

There are a number of people who make an effort such as this

possible. I wish I could thank all who helped not only those whose

suggestions aided the research, but also those who facilitated the

writing.

Included in this group are: Dr. Nikolaus H. Fischer who gave me

encouragement and help when it seemed as if I had reached an

insurmountable obstacle, Dr. Leovigildo Quijano (Professor Instituto de

Quimica, Universidad Nacional de Mexico, Mexico) who was helpful in

discussions about different aspects of structure elucidation, his help

will be always remembered and appreciated. Dr. Francisco A. Macias,

Associate Professor at the Universidad de Cadiz, Spain, who during his

stay at LSU introduced me to the fascination of the sesquiterpene

lactone world, without his enthusiasm and help, I might have never

finished this project; Dr. Lowell E. Urbatsch and Patricia B. Cox to

whom sincere thanks are expressed tofor their aid in plant collection

and identification and for the discussions related to the systematics of

the genus Rudbeckia; and my husband, Robert Zinn, to whom I am grateful

for improving the manuscript as it went through revisions and for

lending computers which made it easier for this dissertation to be

I would like to extend my gratitude to Dr. Luis E. Vidaurreta and

Dr. Robert V. Nauman for their priceless help and encouragement.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. I wish to thank Dr. Frank Fronczek for the X-ray crystallographic

data included in this dissertation.

Although I obtained most of the spectroscopic data in this

dissertation, some infrared spectra were obtained by Rafael Cueto. To

him, I give my thanks.

My thanks too go to Marcus Nauman who helped with the NMR

instrument and to Luz Barona and Henry Hurtado who helped with the

drawings.

Thanks especially are given to my fellow graduate students, present

and former for their help and advice. Their survival of the ordeal gave

me hope. To each of them and others I am grateful.

To these and many other friends, I express my warmest gratitude.

They have all enabled me to do the things I wanted and needed to do.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. This dissertation consists of seven chapters, five of which are

manuscripts that have been published, accepted or submitted for

publication, to various journals. The author of this dissertation, is

also the primary author of the manuscripts, and has received

permission to include the materials in this dissertation. Chapters

1-3 are phytochemical investigations, which concentrate on spectral

interpretation. These chapters were written following the recommended

style for the journal Phytochemistry. Chapters 4 and 5 concentrate on

crystal structures which were written in the style of Acta

Crystallographica. Phytochemistry and Acta Crystallographica are

considered leading international journals in their fields.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CONTENTS OF PART A

ACKNOWLEDGEMENTS...... m

PREFACE...... V

LIST OF TABLES...... viii

LIST OF FIGURES...... x

ABBREVIATIONS...... xvi

DISSERTATION ABSTRACT...... xviii

GENERAL INTRODUCTION...... 1

Chapter 1. SESQUITERPENE LACTONES FROM RUDBECKIA GRANDIFLORA... 7

1.1. Abstract...... 8 1.2. Introduction...... 8 1.3. Results and Discussion...... 9 1.4. Experimental...... 14 References...... 17

Chapter 2. SESQUITERPENE LACTONES AND LIGNANES FROM RUDBECKIA SPECIES AND THE MOLECULAR STRUCTURES OF GAZANIOLIDE AND TAMAULIPIN A ANGELATE...... 51

2.1. Abstract...... 52 2.2 Introduction...... 53 2. 3 Results and Discussion...... 54 2.4 Experimental...... 59 References...... 64

Chapter 3. SESQUITERPENE LACTONES FROM RUDBECKIA MOLLIS...... 90

3.1. Abstract...... 91 3.2. Introduction...... 91 3.3. Results and Discussion...... 92 3.4 Experimental...... 99 References...... 103

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Chapter 4. STRUCTURE OF THE LIGNANE (+)-PINGRESINOL DIMETHYL ETHER...... 126

4. 1 Abstract...... 127 4.2 Experimental...... 127 4. 3 Related Literature...... 129 References...... 130

Chapter 5. STRUCTURES OF THREE SESQUITERPENE y-LACTONES FROM RUDBECKIA MOLLIS...... 136

5.1. Abstract...... 137 5.2. Introduction...... 138 5.3. Experimental...... 138 5.4. Discussion...... 140 References...... 143

Chapter 6. THE SYSTEMATICS OF THE GENUS RUDBECKIA...... 159

6.1. Abstract...... 160 6.2. Introduction...... 160 6.3. Results and Discussion...... 161 A. Taxonomic Implications of Secondary Metabolites in the Genus Rudbeckia...... 161 B. Chemical Distribution...... 162 C. Species Relationship in Rudbeckia...... 163 D. Agricultural, Medicinal, Pest Repellent, and Allelopathic Implications of Biologically Active Secondary Metabolites Present in Rudbeckia...... 165 6.4. Conclusion and Prospects for Future Research.... 166 References...... 170

CONTENTS OF PART B

REVIEW OF SESQUITERPENE LACTONES

Introduction...... 183 References...... 289 Appendix A...... 307 Appendix B...... 315

VITA...... 339

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. LIST OF TABLES

TABLE PAGE

1.1. NMR of compounds 1.1-1. 4...... 18

1.2. NMR data of compounds 1.1-1.4 (50.32 MHz, CDCl^, TMS as internal standard)...... 19

2.1. NMR spectral data of compounds 2.4, and 2.9 (400 MHz, CDCl^, TMS as internal standard)...... 66

2.2. NMR data of compounds 2.4, 2.8 and 2.9 (50.32 MHz, CDCl^, TMS as internal standard)...... 67

2.3. Positional parameters and their estimated standard deviations of compound 2. 4 ...... 68

3.1. ^H NMR spectral data of compounds 3.2, 3.3, 3.4, 3.6 and 3.8 (200 MHz, TMS as internal standard)...... 104

3.2. Coupling constants for compounds 3.2, 3.3, 3.4, 3.6 and 3. 8 in Hertz...... 105

3.3. NMR data of compounds 3.2, 3.3, 3.4, 3.6 and 3.8 (50.32 MHz, CDCl, TMS as internal standard)...... 106

4. 1. Positional parameters and their estimated standard deviations of ( + )-pinoresinol dimethyl ether...... 131

4.2. Bond distances (A) and angles (°) of (+)-pinoresinol dimethyl ether (4.1)...... 132

4.3. Selected torsion angles (°) of (+)-pinoresinol dimethyl ether (4.1)...... 134

5.1. Summary of data collection and structure refinement parameters compounds 5.1-5. 3...... 144

5.2. Position pararameters and their estimated standard deviations compound.5.1...... 145

5.3. Positional parameters and their estimated standard deviations compound.5.2...... 146

5.4. Positional parameters and their estimated standard deviations compound.5.3...... 147

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 5.5. Bond distances in Angstroms compounds 5.1-5..3...... 148

5.6. Bond angles in degrees compounds 5.1-5. 3...... 150

5.7. Selected torsion angles (°) compound 5.1...... 153

6.1. Taxonomy of Rudbeckia...... 168

6.2. Distribution of secondary metabolites inRudbeckia species...... 169

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. LIST OF FIGURES

FIGURE PAGE

1.1. Compounds isolated from floral parts of R. grandiflora... 20

1.2. 100 MHz NMR spectrum of the crude extract of flowers of R. grandiflora in deuteriochloroform...... 21

1.3. Mass spectrum of 6 a-hydroxycostic acid, methyl ester (1.1)...... 22

1.4. Infrared spectra of 6 a-hydroxycostic acid, methyl ester (1.1) and arbusculin E, methyl ester (1.2) (film on KBr window)...... 23

1.5. 400 MHz ^H NMR spectrum of 6 a-hydroxycostic acid, methyl ester (1.1) in deuteriochloroform...... 24

1.6. 400 MHz (^H, ^H) COSY-45 spectrum of 6 a-hydroxycostic acid, methyl ester (1.1). Chemical shifts are given in ppm relative to Me^Si...... 25

1.7. 50.32 MHz ^^C NMR spectrum of 6 a-hydroxycostic acid, methyl ester (1.1)...... 26

1.8. Proton broadband decoupled (BB) and DEPT spectra of 6 a-hydroxycostic acid, methyl ester (1.1) at 50.3 MHz in deuteriochloroform...... 27

1.9. 400 MHz ^H NMR spectrum of 6 a-hydroxycostic acid, methyl ester (1.1) after in situ acylation with TAI 28

1.10. Mass spectrum of arbusculin E, methyl ester (1.2)...... 29

1. 11. 400 MHz ^H NMR spectrum of arbusculin E, methyl ester (1.2) in deuteriochloroform...... 30

1. 12. 400 MHz (^H, ^H) COSY-45 spectrum of arbusculin E, methyl ester (1.2). Chemical shifts are given in ppm relative to Me^Si...... 31

1.13. 50.32 MHz ^^C NMR spectrum of arbusculin E, methyl ester (1.2) in deuteriochloroform...... 32

1. 14 Proton broadband decoupled (BB) and DEPT spectra of arbusculin E, methyl ester (1.2) at 50.3 MHz in deuteriochloroform...... 33

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1.15. 400 MHz NMR spectrum of arbusculin E, methyl ester (1.2) after in situ acylation with TAX...... 34

1.16. Molecular structure of arbusculin E, methyl ester (1.2)...... 35

1.17. Mass spectrum of desacetylligulatin C (1.3)...... 36

1.18. Infrared spectra of desacetylligulatine C (1.3) and rudbeckin A (1.4) (film on KBr window)...... 37

1.19. 400 MHz ^H NMR spectrum of desacetylligulatine C (1.3) in deuteriochloroform...... 38

1.20. 400 MHz (^H, ^H) COSY-45 spectrum of desacetyl ligulatine C (1.3). Chemical shifts are given in ppm relative to Me^Si...... 39

1.21. 50.32 MHz ^^C NMR spectrum of desacetyl ligulatine C (1.3) in deuteriochloroform...... 40

1.22. DEPT spectra of desacetylligulatine C (1.3) at 50.3 MHz in deuteriochloroform...... 41

1.23. Two-dimensional carbon-proton shift correlation of desacetylligulatine C (1.3); ^ C: 50.32 MHz; ^H: 400.1 MHz...... 42

1.24. 400 MHz ^H NMR spectrum of desacetylligulatine C (1.3) after in situ acylation with TAI...... 43

1.25. Mars spectrum of rudbeckin A (1.4)...... 44

1.26. 400 MHz ^H NMR spectrum of rudbeckin A (1.4) in deuteriochloroform...... 45

1.27. 400 MHz (^H. ^H) COSY-45 spectrum of rudbeckin A (1.4). Chemical shifts are given in ppm relative to Me^Si...... 46

1.28. 50.32 MHz ^^C NMR spectrum of rudbeckin A (1.4) in deuteriochloroform...... 47

1.29. DEPT spectra of rudbeckin A (1.4) at 50.3 MHz in deuteriochloroform...... 48

1.30. 400 MHz ^H NMR spectrum of rudbeckin A (1.4) after in situ acylation with TAI...... 49

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1.31. Molecular structure of rudbeckin A (1.4)...... 50

2.1. Compounds isolated from R. grandiflora, R. subtomentosa, R. nitida var. texana and R. maxima...... 70

2. 2. 100 MHz NMR spectrum of the crude extract of leaves of R. subtomentosa in deuteriochloroform...... 71

2.3. 100 MHz ^H NMR spectrum of the crude extract of aerial parts of R. scabrifolia in deuteriochloroform 72

2. 4. 100 MHz ^H NMR spectrum of the crude extract of roots of R. nitida var. texana in deuteriochloroform 73

2.5. 100 MHz ^H NMR spectrum of the crude extract of leaves of R. nitida var. texana in deuteriochloroform...... 74

2.6. 100 MHz ^H NMR spectrum of the crude extract of roots of R. subtomentosa in deuteriochloroform. Spectrum provided by Dr. Leovigildo Quijano...... 75

2.7. 100 MHz ^H NMR spectrum of the crude extract of roots of R. grandiflora in deuteriochloroform...... 76

2.8. Infrared spectra of tamaulipin A angelate (2.4) (film on KBr window)...... 77

2.9. 400 MHz ^H NMR spectrum of tamaulipin A angelate (2.4) in deuteriochloroform...... 78

2. 10. 400 MHz (^H, ^H) COSY-45 spectrum of tamaulipin A angelate (2.4). Chemical shifts are given in ppm relative to Me^Sl...... 79

2.11 2D J-Resolved spectrum of tamaulipin A angelate (2.4).... 80

2.12. 50.32 MHz ^^C NMR spectrum of tamaulipin A angelate (2.4) in deuteriochloroform...... 81

2.13. Proton broadband decoupled (BB) and DEFT spectra of tamaulipin A angelate (2.4) at 50.3 MHz in deuteriochloroform...... 82

2.14. Two-dimensional carbon-proton shift correlation of tamaulipin A angelate (2.4); ^^C: 50.32 MHz; H: 400.1 MHz...... 83

2.15. Circular Dichroism spectrum of tamaulipin A angelate (2.4) in methanol...... 84 xii

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 2.16. Single crystal X-ray structure of tamaulipin A angelate (2.4) 85

2.17. 400 MHz NMR spectrum of isogazaniolide (2.9) in deuteriochloroform. Spectrum provided by Dr. Leovigildo Quijano...... 86

2. 18. 400 MHz (^H, ^H) COSY-45 spectrum of isogazaniolide (2.9). Chemical shifts are given in ppm relative to Me^Si. Sample provided by Dr. Leovigildo Quijano...... 87

2.19. 50.32 MHz ^^C NMR spectrum of isogazaniolide (2.9) in deuteriochloroform. Spectrum provided by Dr. Leovigildo Quijano...... 88

2.20. DEPT spectra of isogazaniolide (2.9) at 50.3 MHz in deuteriochloroform. Sample provided by Dr. Leovigildo Quijano...... 89

3.1. Compounds isolated from Rudbeckia mollis...... 108

3.2. 100 MHz ^H NMR spectrum of the crude extract of leaves of Rudbeckia mollis in deuteriochloroform...... 109

3.3. 100 MHz ^H NMR spectrum of the crude extract of flowers of Rudbeckia mollis in deuteriochloroform...... 110

3.4. 100 MHz ^H NMR spectrum of the crude extract of aerial parts of Rudbeckia mollis in deuteriochloroform...... Ill

3.5. 400 MHz ^H NMR spectrum of llaH,13-dihydrorudmollin (3. 2) in deuteriochloroform...... 112

3.6. 400 MHz (^H, ^H) COSY-45 spectrum of llocH, 13-dihydrorudmollin (3.2). Chemical shifts are given in ppm relative to Me^Si...... 113

3.7. 50.32 MHz ^^C NMR spectrum of llaH,13-dihydrorudmollin (3.2 )...... 114

3.8. Proton broadband decoupled (BB) and DEPT spectra of llaH,13-dihydrorudmollin (3.2) at 50.3 MHz in deuteriochloroform...... 115

3.9. 400 MHz ^H NMR spectrum of llaH,13-dihydrorudmollin (3.2) after In situ acylation with TAI...... 116

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 3.10. Circular Dichroism spectrum of llctH,13-dihydrorudmollin (3.2) in methanol...... 117

3.11. 400 MHz NMR spectrum of lloH,13-dihydrorudmollin diacetate (3.2) in deuteriochloroform...... 118

3.12. 400 MHz ^H NMR spectrum of lloH,13-dihydrorudmollin diacetate (3.4) in deuteriobenzene...... 119

3.13. 400 MHz (^H, ^H) COSY-45 spectrum of llaH,13-dihydrorudmollin diacetate (3.4). Chemical shifts are given in ppm relative to Me^Si...... 120

3.14 2D J-Resolved spectrum of lloH,13-dihydrorudmollin diacetate (3.4)...... 121

3.15. 50.32 MHz ^^C NMR spectrum of llaH,13-dihydrorudmollin diacetate (3.4)...... 122

3. 16. Proton broadband decoupled (BB) and DEPT spectra of llaH,13-dihydrorudmollin diacetate (3.4) at 50.3 MHz in deuteriochloroform...... 123

3.17. Circular Dichroism spectrum of llaH,13-dihydrorudmollin diacetate (3.4) in methanol...... 124

4.1. Single crystal X-ray structure of (+)-pinoresinol dimethyl ether (4.1)...... 136

5.1. The molecular structure of 15-acetylrudmollin (5.1), with thermal ellipsoids drawn at the 30% probability level, and H atoms represented by circles of arbitrary

5.2. The molecular structure of diacetylrudmollin (5.2), with 30% ellipsoids...... 157

5.3. The molecular structure of llaH,13-dihydrorudmollin (5.3), with 50% ellipsoids...... 158

6.1. Carotenoids isolated from Rudbeckia species...... 172

6.2. Flavonoids isolated from Rudbeckia species...... 173

6.3. Polyacetylenes isolated from Rudbeckia species...... 174

6.4. Sesquiterpene lactones isolated from Rudbeckia mollis.... 175

6.5. Secondary metabolites isolated from roots of Rudbeckia laciniata...... 176

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 6.6. Secondary metabolites isolated from aerial parts of Rudbeckia laciniata...... 177

6.7. Secondary metabolites isolated from Rudbeckia fulgida.... 178

6.8. Geographical distribution of Rudbeckia triloba in the southeastern United States...... 179

6. 9. Geographical distribution of Rudbeckia laciniata in the southeastern United States...... 180

6. 10. Geographical distribution of Rudbeckia fulgida and Rudbeckia grandiflora in the southeastern United States...... 181

6.11. Geographical distribution of Rudbeckia mollis and Rudbeckia subtomentosa in the southeastern United States...... 182

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. PRINCIPAL ABBREVIATIONS

broadband cc Column chromatography CD Circular dichroism COSY Correlation Spectroscopy 2D two dimensional DEPT Distorsionless Enhancement by Polarization Transfer eV electron volts FAB Fast Atom Bombardment HRMS High Resolution Mass Spectroscopy Hz Hertz (cycies per second) IR Infrared Low Resolution Mass Spectroscopy Molecular ion MHz megahertz MS Mass spectrometry NMR Nuclear Magnetic Resonance NOE Nuclear Overhauser Effect TLC Thin-layer chromatography UV Ultraviolet spectroscopy

various

doublet Chemical shift in ppm relative to TMS Coupling constant in hertz chemical shift increment multiplet quartet singlet

wavelength Mass-to-charge ratio

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Ac^O Acetic anhydride deuterobenzene CHCl^ chloroform EtOAc Ethyl acetate MeOH Methanol NaBH Sodium tetrahydroboride petrol petroleum ether TAG trichloroacetylcarbamate TAI trichloroacetyl isocyanate TMS tetramethylsilane

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. DISSERTATION ABSTRACT

This dissertation consists of two parts. In part A, the isolation

and structure elucidation of secondary metabolites from the genus

Rudbeckia are described. A comprehensive review of newly reported

sesquiterpene lactones published during 1986 and 1987 is presented in

As part of a biochemical systematic study within the family

Asteraceae, seven species of the genus Rudbeckia were investigated for

secondary terpenoid metabolites. This study resulted in the isolation of

42 compounds, of which 10 are new natural products. The compounds

include sesquiterpene esters, pseudoguaianolides, germacrolides,

eudesmano1ides, lignanes and flavonols among others. The new compounds

are 6a-hydroxycostic acid, methyl ester, arbusculin E, methyl ester,

desacetylligulatine C, rudbeckin A, tamaulipin A angelate,

llaH,13-dihydrorudmollin, 15-acetyl-llaH,13-dihydrorudmollin,

4-acetyl-llaH,13-dihydrorudmollin, 15-acetylconfertin and

llaH,13-dihydrorudmollin diacetate. Their structures were established by

means of chemical and spectroscopic methods. These methods included MS,

IR, UV, CD, ^H NMR and NMR. Single crystal X-ray diffraction

analysis of the following compounds confirmed the proposed structures:

(+)-pinoresinol methyl ether, tamaulipin A angelate, 15-acetylrudmollin,

rudmollin diacetate and llaH,13-dihydrorudmollin.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. The comprehensive bibliographic review provides information about

522 new sesquiterpene lactones reported in the literature from the

beginning of 1986 through the end of 1987. New plant sources (174) for

sesquiterpene lactones are listed in this review which consists of a

primary table which is arranged on the basis of structural types and

substitution patterns.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. GENERAL INTRODUCTION

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. The first section of this dissertation consists of five chapters

describing the structure elucidation of natural products. The first

three chapters, written for publication in Phytochemistry report the

isolation and structure elucidation of secondary metabolites from five

species of the genus Rudbeckia. Chapters 4 and 5, also prepared for

publication, report X-ray diffraction analyses. Chapter 6, summarizes my

results, and contains recommendations for future work. The second, and

final section, consists of a complete review of the literature on new

sesquiterpene lactones, published in the years 1986 and 1987. I plan to

publish this review as a book.

In chapter 1, I include information from the chemical analysis of an

extract from flowers of Rudbeckia grandiflora. This study provided two

new sesquiterpene esters, 6a-hydroxycostic acid, methyl ester and

arbusculin E, methyl ester as well as two new pseudoguaianolide <-ype

sesquiterpene lactones, desacetylligulatine C and rudbeckin A.

In chapter 2, the results of chemical studies of five Rudbeckia

species for their secondary metabolites are presented. Taxa sampled were

R. grandiflora, R. subtomentosa, R. nitida var. texana, R. scabrifolia

and R. maxima. The leaves of R. subtomentosa afforded the known

sesquiterpene lactones 4-0-desacetylligulatine C, and ligulatine C;

flowers provided costunolide, santamarine, reynosin, the flavonol

eupatolin, plus a mixture of g-sitosterol and stigmasterol; roots gave

costunolide, 11,13-dihydrocostunolide, gazaniolide, isogazaniolide.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. tamaulipin A angelate, caryophyllene, caryophyllene oxide, and

p-selinene. Extracts from R. nitida var.texana, R. scabrifolia, and R.

maxima gave the lignanes (+)-pinoresinol dimethyl ether and yangambin.

The germacrolide tamaulipin A angelate was the major constituent of root

extracts of R. grandi flora. Structure and stereochemistry of the new

lactones were elucidated by spectroscopic methods. The molecular

structures of gazaniolide and tamaulipin A angelate were established by

single crystal X-ray diffraction.

11,13-Dihydrocostunolide, gazanolide, isogazanolide, caryophyllene,

caryophyllene oxide and g-selinene, which were isolated from roots of R.

subtomentosa, were identified by Dr. Leovigildo Quijano. Dr. Quijano’s

results were included in the manuscript because the compounds obtained

by Dr. Quijano were closely related to those I had isolated from the

same species. Since the data enhanced the par:.r, they are included in

the chapter, but are not presented nor claimed as work done by the

author of this dissertation, except in as much as this author integrated

that information into the structure of the paper.

Chapter 3 contains the structure and stereochemistry of four new

pseudogua1ano1Ide type sesquiterpene lactones from Rudbeckia mollis. The

structures of llaH,13-dihydrorudmollin, 15-acetyl-llocH,13-

dihydro-rudmol1in, 4-acetyl-llocH,13-dihydrorudmollin and

15-acetylconfertin were determined from spectroscopic data and chemical

t- ansformations. The new pseudoguaiano1ide lltxH,13-dihydrorudmollin

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. diacetate was prepared. Data on the known compounds rudmollin, rudmollin

diacetate, 15-acetylrudmoIlin, 4-acetylrudmollin, and eupatolin are also

reported.

Chapter 4 describes the crystal structure of the lignane

(+)-pinoresinol dimethyl ether. The compound was isolated from Rudbeckia

maxima Nutt, R. nitida Perdue and R. scabrifolia. It was found by Dr.

Fronczek that crystals of the title compound are orthorhombic. The two

five-membered rings of the central dioxabicyclooctane system are

cis-fused; each ring adopts the half-chair conformation in which one

atom lies on both pseudodiads. The phenyl rings are planar and the four

methoxy substituents lie near these planes.

Chapter 5 includes the X-ray data of three ambrosanolide-type

pseudoguaianolides compounds isolated from Rudbeckia mollis.

Thestructures of three compounds were resolved by Dr. Fronczek. These

showthe seven-membered rings in a twist-boat conformation, with CIO on

the pseudodiad and the cyclopentane ring is in an envelope conformation

in which C5 is the flap. Hydroxyl groups in both compounds are involved

in hydrogen bonding.

In chapter 6, I summarize the present state of knowledge of the

chemistry of the genus Rudbeckia. Sesquiterpene lactones, polyacetylenes

and lignanes are typical constituents in Rudbeckia species; there are

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. many structural variations among the lactones. Various sesquiterpene

lactones and lignanes obtained from Rudbeckia are known to show

antitumor activity and to inhibit certain enzymes.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Copyright Administrator Journals Division Pergamon Press pic, Headington Hill Hall, Oxford 0X3 OBW, England

Dear Sir:

As the primary author of the article "Sesquiterpenes from Rudbeckia grandiflora" published in Phytochemistry, 1988, 17, 2195-2198. I hereby request permission to use the materials, in whole, or in part, as a portion of my Ph.D. dissertation.

I would appreciate your granting of this permission as promptly as possible

Marta Vasquez Box E-4, Choppin Hall Department of Chemistry Louisiana State University Baton Rouge, LA 70803

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. SESQUITERPENE LACTONES FROM RUDBECKIA GRANDIFLORA

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. (reprinted with permission from Phytochemistry, 1988, 17, 2195-2198. Copyright © 1989 Pergamon Press)

Marta Vasquez, Francisco A. Macias, Lowell E. Urbatsch and Nikolaus H. Fischer

Department of Chemistry;^Department of Botany, Louisiana State University, Baton Rouge, LA 70803, U.S.A.

(Received 29 September 1987)

Key Word Index-Rudbeckia grandiflora; Asteraceae; Heliantheae; Sesquiterpene esters; eudesmanes; sesquiterpene lactones; pseudogua iano1ides.

ABSTRACT-Chemical analysis of Rudbeckia grandiflora afforded two new sesquiterpene esters, 6a-hydroxycostic acid, methyl ester and arbusculin E, methyl ester as well as two new pseudoguaianolide type sesquiterpene lactones, desacyl1igulatine C and rudbeckin A. Their structures were elucidated by spectroscopic methods and chemical transf ormat ions.

INTRODUCTION

The genus Rudbeckia L. consists of approximately 18 species and

30 taxa of annual and perennial herbs divided into two subgenera

Rudbeckia and Macrocllne. Rudbeckia grandiflora is very abundant

along roadside and in pastures in Arkansas, Oklahoma and Louisiana.

Previous chemical studies of members of this genus included R. mol Hi

[1] and R. laclnlata [2]. Sesquiterpene lactones isolated from R.

mollis exhibited antitumor activity [1].

As part of a biochemical systematic study within the tribe

Permanent address: Departamento de Quimica Organica, Facultad de Ciencias, Universidad de Cadiz, Apdo 40, 11080 Puerto Real, Cadiz,

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Heliantheae combined with a search for bioactive plant products we

have analyzed the previously uninvestigated species R. grandiflora.

The structures of the four new compounds which we named

6a-hydroxycostic acid, methyl ester, (1) arbusculin E methyl ester,

(2) desacyl1igulatine C, (3) and rudbeckin A (4) were established by

chemical and spectroscopic methods.

RESULTS AND DISCUSSION

The dichloromethane extract of floral parts of R. grandiflora,

afforded after column chromatography and further purification by

preparative TLC, four new compounds. 6a-Hydroxycostic acid, methyl

ester (1), exhibited in the NMR spectrum a three proton

singlet at 50.82 suggesting an angular methyl in an eudesmane

skeleton. Two one proton broadened singlets at 56.28 and 5.22

together with three proton singlet at 53.79 indicated an «-methylene

methyl ester of the costic acid type [3]. Two broadened singlets at

55.00 and 4.67 were in agreement with an olefinic methylene group

which had to be on C-15 on biogenetic grounds. A doublet of doublets

at 53.92 was assigned to a proton on a carbon bearing a hydroxyl

group. It’s coupling to H-5, a doublet at 51.91 (J=10.1 Hz), and to

H-7 (ddd, 52.60) as determined by 20 COSY experiments. This finding

unambiguously established the position of the hydroxyl group at C-6.

The stereochemistry at C-6 was derived from the couplings between

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. trans-di-axlal orientation of these three protons. Assuming an H-7 a

as in all sesquiterpenes from higher plants [4] the hydrogens at C-6

and C-5 had to be a and g respectively. This was further

substantiated by NOE difference experiments which showed effects

between H-5 and H-7 but had no effect on H-6. In situ acylation of

the hydroxyl group with trichloroacetyl isocyanate (TAI) [5]

corroborated the above findings. The ^H NMR spectrum of the

trichloroacetylcarbamate (TAG) derivative (la) showed one NH signal

at 58.20, providing further evidence for the presence of a hydroxyl

group in compound 1. The paramagnetic acylation shift of the doublet

of doublets at 53.92 (H-6) in 1 to 55.5 in la (A5=l.58) was

consistent with a geminal position of H-6 and the hydroxyl group. The

signal of the C-10-methyl group at 50.82 showed no paramagnetic

shift, implying that the C-10 -methyl is oriented away from H-6,

which is in agreement with the relative stereochemistry of the two

groups, that is, C-6 a-hydroxyl and C-10 p-methyl. All other proton

absorptions of ester 1 were assigned on the basis of 2D COSY, spin

decoupling and NOE difference experiments (Table 1). The ^^C NMR

spectrum of 1 (Table 2) was assigned with the aid of the

heteronuclear multipulse DEPT experiments; the data of this new

sesquiterpene supported the ^H NMR assignments of 1 which was named

6a-hydroxycostic acid, methyl ester.

Arbusculin E, methyl ester (2) C

(Table 1) values are very similar to those of 6a-Hydroxycostic acid.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 11

methyl ester (1 ), the major differences between 1 and 2 residing in

the proton signals of C-4/C-15. Instead of the two olefinic protons

at C-15 in compound 1 compound 2 exhibited a three protons singlet at

Ô1.32, which is characteristic of a methyl group on a carbon bearing

a hydroxyl group. The chemical shift of H-5 in 2 was also shifted

upfield by about 0.5 ppm suggesting that it was in a non-allylic

position. Therefore the methylene group on C-4 in 1 had to be changed

to a methyl group at a carbon bearing a hydroxyl group in 2 as

determined by 2D COSY experiments. The ^^C NMR spectrum of 2 (Table

2) corroborated the proposed structure with signals at 5106.92 (C-15)

and 14.3 (C-4) in the spectrum of 1 being replaced by signals at

523.78 (C-15) and 73.47 (C-4) in compound 2. The ^^C NMR spectrum of

2 was assigned with the aid of the heteronuclear multipulse DEPT

experiments.

The stereochemistry of the chiral centers was proposed on the

basis of the results of the TAI experiment and by examination of

stereomodels. The NMR spectrum of the trichloroacetylcarbamate 2b

showed NH signals at 58.20 and 58.30 indicating two hydroxyl groups

in compound 2. The signal of H-6 at 54.10 in the spectrum of compound

2 was shifted to 55.28 in that of 2b (A5=l.18) due to the strong

deshielding effect of the TAG acyl group. Also, the C-4-methyl group

was shifted from 51.32 in the spectrum of 2 to 51.60 in that of 2b

(A5=0.28). These shifts strongly suggest the a-orientation of the two

hydroxyl groups in the molecule as well as the «-orientation of H-5.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 12

The structure of 2 was unambiguously established by its

transformation to 6a-hydroxy costic acid (2a) with spectral NMR data

being in complete agreement with data reported in the literature [3].

Compound 2 which was not previously reported as a natural product,

represents the methyl ester of the known arbusculin E [7].

Desacyl 1 igulat ine C (3), is a gum with NMR spectral

signals typical for an a-methylene-y-lactone moiety with two

one-proton doublets at 56.20 (H-13a) and 5.49 (H-13b), both coupled

to a one-proton multiplet at 53.45 (H-7). Strong IR absorptions at

1757 and 3429 cm ^ corroborated the presence of a y-lactone and

hydroxyl(s), respectively. 20 COSY studies indicated that H-7 was

coupled to a doublet at 54.51 (J^ ^=9.5 Hz) which was assigned to the

lactonic proton at C-6. The multiplicity of H-6 suggested a

pseudoguaianolide-type skeleton for lactone 3 [4]. A three proton

doublet at 50.99 (J=7. 5 Hz) was assigned to the methyl group at C-10.

However, the common methyl signal due to an angular methyl at C-5,

was missing in compound 3. Instead, a pair of geminally coupled

doublets (J=12.2 Hz) at 54.05 and 3.90 suggested the presence of a

-CH^OH group at C-5, which was supported by a ^^C NMR triplet at

561.82. Compound 3 lacked a ^^C NMR absorption for a cyclopentanone

carbonyl but exhibited a doublet at 583.91 typical of a carbon

bearing an oxygen. The ^H NMR spectrum supported the above findings

by the presence of a doublet of a doublet at 54.35 (J=8.8 and 9.0

Hz). The chemical shift together with the multiplicity of this proton

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. suggested the presence of a hydroxyl group at C-4. All other proton

signals were assigned on the basis of spin decoupling experiments, 2D

COSY and 2D correlations (Table 1). The ^^C spectral signals

(Table 2) were assigned on the basis of 2D COSY, correlation

and DEPT experiments. The spectroscopic data were consistent with

data reported for a synthetic derivative of ligulatine C [6]. To our

best knowledge this is the first report of desacylligulatine C [3] as

a natural product.

Rudbeckin A (4), showed an IR band at 1765 cm ^

(y-lactone) and the NMR (Table 1) also suggested a lactone which

had to be closely related to desacyl1igulat ine C (3). Strong IR

absorptions at 1765 and 3418 cm”^ corroborated the presence of a

y-lactone and hydroxyl(s) respectively. Compound 4 differed from 3 in

that the two doublets characteristic of an a-methylene-y-lactone were

missing in spectrum of compound 4. Instead, a three-proton doublet at

61.35 (J=7.8 Hz) appeared which was assigned to the C-11 methyl

group. The doublet at 60.99 (C-ll-Me) in the spectrum of 3 was

replaced by a singlet at 61.3 characteristic of a methyl group at a

carbon bearing a hydroxyl an H-7 at 62.25 appeared to be non-allylic.

The ^^C NMR spectrum (Table 2) was assigned by comparison of 4 with

lactone 3. Application of heteronuclear multipulse DEPT experiments

and 2D COSY allowed assignment of all ^H NMR spectral signals.

The above spectral data were in agreement with structure 4

exclusive of its stereochemistry, which was determined with the aid

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 14

of NOE difference spectral data and TAI experiments. Irradiations of

the C-11-methyl signal from NOE Difference experiment showed an

effect on H-6 and on H-7, indicating an a-orientation of the

C-11-methyl group. The ^H NMR spectrum of the TAG derivative (4a),

showed a paramagnetic acylation shifts of H-4 (Aô=1.16), H-15

(Aô=1.35) and H-1 (Aô=0.20) establishing the presence of hydroxyl

groups at C-4 and C-15 and the a-orientation for the hydroxyl group

on C-10. An axial p-hydroxyl group should have caused a noticeable

shift of the H-9a and H-2p signals, which was not observed.

C-11-methyl signal was not shifted significantly which is consistent

with an a-orientation for the C-ll-methyl group. Similarly the H-6a

signal was not affected suggesting a p-orientation of the CH^OH group

at C-5. This new pseudoguaianolide has not been previously reported

and is named rudbeckin A (4).

EXPERIMENTAL

Rudbeckia grandiflora (Sweet) DC. was collected on 7 June 1986,

in pine woods and roadside of Louisiana 121, 3.3 miles west of

Rapides Parish line east of La Camp, sec 16, I 2N,R5W. (P. Cox, L.

Urbatsch and E. Harris No. Cox 4889, voucher deposited at L.S.U.,

U.S.A. ).

The air-dried flowers (50 g) were ground and extracted according

to the general procedure [8] providing 1.3 g of the crude terpenoid

extract. The crude extract (1.0 g) was separated by CC on silica gel

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. using hexane-EtOAc mixtures of increasing polarity, 86 fractions of

50 ml each being collected. Upon further prep. TLC of the various

fractions two sesquiterpenes (1 and 2) and two sesquiterpene lactones

(3 and 4) were isolated. Fraction 12 provided 30 mg of 1 and

fractions 24, 25 gave 34 mg of 2. Fractions 29-31 afforded 3 (50 mg)

and fractions 40, 41 provided 4 (35 mg).

6a-hydroxycostic acid, methyl ester

IR cm"\ 3511 (OH), 1719 (ester), 1647 and 1626 (double bond);

HRMS (FAB probe) 70 eV, m/z (rel. int.): 264.1247 [M]* (4) (calc, for

^16^24°3' 264.1249), 249.1330 [M-Me]* (2), 233.1145 [M-DMe]* (2),

186.0713 (64), 185.0781 (96), 93.0417 (100); NMR see Table 1.

see Table 2; TAC-derivative of 1 NMR H-6 d, 63.92 (A6=+1.58).

Arbusculin E, methyl ester (2). colorless gum; IR

cm"\ 3347 (OH), 1719 (ester) 1626 and 1439 (double bond) HRMS (FAB

probe) 70 eV, m/z 282.1541 [M]* (37) (calc, for C^^H^^O^: 282.1544),

265.1917 (75), 251.1692 [M-OMe]* (8), 247.1750 (37), 233.1542 (74),

161.1285 (48), 133.8546 (100), 109.0871 (45). ^H NMR see Table 1.

see Table 2; TAC-derivative of 2: ^H NMR H-6, 64.10 (A6=+l.18); H-4,

61.32 (A6=+0.28).

Saponification of compound 2. A 10 mg sample of 2 was dissolved

in 3 ml of 5% methanolic KOH. After 16 hr the soln was diluted with

H^O (10 ml) and neutralized with 1% HCl. Ex

(5 mg) which was purified by prep. TLC [3].

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1757 (y-lactone), 1663 (double bond). HRMS (FAB, probe) 70 eV m/z

266.1434 [Ml* (3), (calc, for 266.1434), 248.1377 [M-H^O]*

(6), 230.1105 [M-2H^0]* (16.7) 235.1268 [M-CH^OH]* (19), 203.1301

(68), 159.1097 (100), 147.0913 (83), 145.0772 (94), 105.0594 (88),

95.0637 (84). NMR see Table 1; NMR see Table 2. TAC-derivative

of 3. NMR H-15', Ô3.90 (A0=+0.62), H-15, 64.05 (A6=+0.55), H-4,

54.35 (A5=+0.69), H-6, 54.51 (A5=+0.99).

Rudbeckin A (4). C^gH^^O^, gum; IR cm“S 3418 (OH), 1765

(y-lactone). HRMS (FAB, probe) 70 eV m/z: 266.1614 [M-H^O]* (26)

(calc, for [M-H^O]*: 266.1616), 265.1161 (76), 251.1371

(74), 249.1264 (82), 231.1502 (M-2H^0) (30) 219.1437 [M-3H^0] (31),

203.1137 (62), 185.0731 (94), 93.0718 (100); ^H NMR see Table 1.

NMR see Table 2. TAC-derivative of 4: ^H NMR H-4, 4.24 (A5=+l. 16),

H-15, 54.05 (A5=+1.35), H-1, 52.25 (A5=+0.20)

Acknowledgements-The authors are grateful to the U.S.-Spain Joint

Committee for Scientific and Technological Cooperation (Project No

CCB-8409023) for support of this research and F.A.M. for funds

allowing a visiting professorship at Louisiana State University

(Project No IPB-850911). Purchase of 100 and 400 MHz NMR

spectrometers was made possible by NIH Shared Instrumentation Grant 1

SIO RR02459-01.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1. Herz, W. and Kumar, N. (1981) J. Org. Chem. 46, 1356.

2. Bohlmann, F., Jakupovic, J. and Zdero, C. (1978) Phytochemistry 17. 2034.

3. Herz, W., Chikamatsu, H. and Tether, L. R. (1966) Phytochemistry 18, 1189.

4. Fischer, N. H., Oliyier, E. J. and Fischer, H. D. (1979) in Progress in the Chemistry of Organic Natural Products (Herz. W., Grisebach, H. and Kirby, G. B. , eds). Springer, Vienna.

5. Samek, K. and Budesinsky, M. (1979) Collect. Czech. Chem. Comm. 44, 558.

6. Maldonado, E., Mendoza, G. 0., Cardenas, J. and Ortega, A. (1985) Phytochemistry 24, 2981.

7. Asakawa, Y., Ourisson, G. and Aratani, T. (1975) Tetrahedron Letters 45, 3957.

8. Fischer, N. H., Wiley, R. A., Lin. H. N. , Karimian, K. and Politz, S. M. (1975) Phytochemistry 14, 2241.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. z.CDCl*. THS #. inwrnml .tmndmrd)»

-.2o«Ifl.2««7.0Ù-8A.9fl-3.5

• Th« signals du# to hydroxyl group# ai t multiplicity and coupling c * Obflcur#d by othar signals.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.2. C NMR data of compounds 1-4

(50.32 MHz, CDCl , IMS as internai standard)

c 1 2 3+ 4+

1 41.86t* 42.92t * 48.45d 46.89d

2 19.53t 22.30t 22.69t

3 29.70t 27.let

4 73.47s 83.91d 82.74d

5 57.92d 57.92d 53.52s 60.23s

6 69.33d 73.26d 90.09d 85.71d

7 48.07d 50.25d 43.44d 43.99d

8 23.93t 26.80t 32.29t 31.86t

9 40.38t* 42.63t* 24.80t 43.41t

10 37.40s 32.76d 85.31s

147.27s 139.88s 53.OOd

12 168.33s 179.45s

13 124.63t 125.75t 118.89t 16.64q

14 17.61q 19.70q 13.95q 24.32q

15 106.92t 23.78q 61.82t 68.91t

OMe 51.84q 52.01q - -

Peak multiplicity was obtained by heteronuclear multipulse programs. ^ Assignments for 3 and 4 were confirmed by chemical shift correlation. t Assigments may be interchanged

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. k.6>x)i>C0,Me

1 : R = H 2 :R = R'=H;R=cH3 la : R = C0NHC0CCl3 2a: R= R'=R"=H 2b: R= R' = C0NHC0CCl3

R O O R

3 : R = R'= H 4 :R=R'=R"=H 3 a : R = R'=C0NHC0CCl3 4a:R=R'=R=C0NHC0CCl3

Fig. 1.1. Compounds isolated from floral parts of R. grandiflora.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1.2. 100 MHz NMR spectrum of the crude extract of flowers of R. grandiflora in deuteriochloroform.

8.0 6.0 4.0 2.0 0.0 PPM

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 1.3. Mass spectrum of 6 a-hydroxycostic acid, methyl ester (1.1).

265 309

\'f

llii150 200 250 300 350

20.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 4B8.B 1.B412

1.950

OH OH

4000

B.967 = = -

8.9 8 J

).B0^

C i ; 0 . 5 8 - j

4000

Fig. 1.4. Infrared spectra of 6 a-hydroxycostic acid, methyl ester (1.1) and arbusculin E, methyl ester (1.2) (film on KBr window).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. II It •HOO

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 1.6. 400 MHz (*H, ^H) COSY-45 spectrum of 6 a-hydroxycostic acid, methyl ester (1.1).

OCH-

13,0 13b, 5 ^ ,5b

2.0

3.0

■4.0

5. 0

6.0

■8.0 7.0 6.0 5.0 40 3.0 2.0

*Chemical shifts are given in ppm relative to Me^Si.

13 15 5 x

II4 wwV

1 8 0 1 4 0 100 6 0 20 PMM Fig. 1.8. Proton broadband decoupled (BB) and DEPT spectra of 6 a-hydroxycostic acid, methyl ester (l.J) at 50.3 MHz in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. JA ilL ak 7 6 5 4 3 2 1 0

7 5

ppm

Fig. 1.9. 400 MHz H NMR spectrum of 5 a-hydroxycostic acid, methyl ester (1.1) after in situ acylation with TAX.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 4®.

Fig. 1.10. Mass spectrum of arbusculin E, methyl ester (1.2).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ; 2

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 1. 12. 400 MHz (^H, ^H) COSY-45 spectrum* of arbusculin E, 31 methyl ester (1.2).

Chemical shifts are given in ppm relative to Me Si.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 1.14 Proton broadband decoupled (BB) and DEPT spectra of arbusculin E, methyl ester (1.2) at 50.3 MHz in deuteriochloroform.

6 • 5 D e p t 9 0 LLI

D e p t l 3 5

1 6 0 140 120 100 80 6 0 4 0 20 ppm

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. OH OH

13b 1 3 a

13b 1306

.11

13a 13b 6 J 1 1 A-A.

ppm 8 6 4 2 0

Fig. 1.15. 400 MHz NMR spectrum of arbusculin E, methyl ester (1.2) after in situ acylation with TAI.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 1.17. Mass spectrum of desacetylligulatin C (1.3).

1 1 jjj L |ln.lil|lilJ 41--I 250

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1.8255

1.900

2880 1888

0 .7 0-

3 .6 8-

4800 1088

Fig. 1.18, Infrared spectra of desacetylligulatine C (1.3) and rudbeckin A (1.4) (film on KBr window).

6.0 5.0 4.0 3.0 2.0

Fig. 1.20. 400 MHz C H, H) COSY-45 spectrum of desacetylligulatine C (1.3). Chemical shifts are given in ppm relative to He Si.

1 6 0 120 8 0 4 0

Fig. 1.22. DEPT spectra of desacetylligulatine C (1.3) at 50.3 MHz in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibitedpermission. without J

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 14 43 1 5 b

13a 13b 15o 'U

15b

13c '3b

7 6 5 4 3 2 1 PPM

Fig. 1.24. 400 MHz NMR spectrum of desacetylligulatine C (1.3) after Jn situ acylation with TAI.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 60.

20.

Fig. 1.25. Mass spectrum of rudbeckin A (1.4).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. SJ T

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 1.27. 400 MHz (*H, ^H) COSY-45 spectrum of rudbeckin A (1.4). 14

1 5 b ISal

2.0

■3.0

-5.0

4.0 3.05.0 2.0 1.0

Chemical shifts are given In ppm relative to Me^Si.

D e p t l 3 5

_jJ I D e p t 9 0

80 60 40 20 0 PPM

Fig. 1.29. DEPT spectra of rudbeckin A (1.4) at 50.3 MHz in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 1.30. 400 MHz NMR spectrum of rudbeckin A (1.4) after in situ acylation with TAI.

ppm 7 6 5 4 3 2 1

SESQUITERPENE LACTONES AND LIGNANES

FROM RUDBECKIA SPECIES AND THE MOLECULAR STRUCTURES

OF GAZANIOLIDE AND TAMAULIPIN A ANGELATE

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. (submitted to Phytochemistry)

Marta Vasquez, Leovigildo Quijano, Frank R. Fronczek, Francisco A.

Macias? Lowell E. Urbatsclf, Patricia B. Coj^, and Nikolaus H. Fischei^ Department of Chemistry and ^Department of Botany

Louisiana State University, Baton Rouge, LA 70803, U.S.A.

(Received )

Key word Index-Rudbeckla; Asteraceae; Heliantheae; sesquiterpene lactones; pseudoguaiano1ides; germacrolides; eudesmanolides; lignanes; crystal structures.

Abstract-Five Rudbeckia species (R. grandiflora R. subtomentosa, R. nitida var. texana, R. scabrifolia and R. maxima) were investigated for their secondary constituents. R. subtomentosa leaves afforded the known sesquiterpene lactones 4-0-desacetylligulatine C, and ligulatine C; flowers provided costunolide, santamarine, reynosin, the flavonol eupatolin plus a mixture of g-sitosterol and stigmasterol; roots gave costunolide, 11,13-dihydrocostunolide, gazaniolide, isogazaniolide, tamaulipln A angelate, caryophyllene, caryophyllene oxide and g-selinene. R. nitida var. texana, R. scabrifolia and R. maxima gave the lignanes (+)-pinoresinol dimethyl ether and yangambin. The germacrolide tamaulipln A angelate, was the major constituent of root extracts of R. grandiflora. Structure and stereochemistry of the new lactones were elucidated by spectroscopic methods. The molecular structures of gazaniolide and tamaulipln A angelate were established by single crystal X-ray diffraction.______

Permanent address: Institute de Quimica, Universidad Nacional Autonoma de México, México. 20, D. F. ^Permanent address Departamento de Quimica Organica, Facultad de Ciencias, Universidad de Cadiz, Apdo. 40, 11080, Puerto Real Cadiz, Spain. ^Author to whom correspondence should be addressed

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. INTRODUCTION

In continuation of our biochemical systematic study of the tribe

Heliantheae (Asteraceae) combined with a search for bioactive natural

plant products we have investigated five species of the genus

Rudbeckia L. Previous chemical analyses of members of this genus were

performed on R. mollis [1], R. laciniata [2], R. grandi flora [3], R.

hirta [4-7], R. triloba [8] and R. fulgida [9]. Flavonoids [1,4,5,9],

polyacetylenes [6-9], carotenoids [10], eudesmane-type sesquiterpene

esters [3] and pseudoguaianolides [1,3] were found in these

investigations.

Since "hairy root" cultures obtained upon transformation with

Agrobacterium rhizogenes have been shown to express the pattern of

secondary metabolites characteristic of the species from which they

are derived [11], we have also analyzed the roots of several

Rudbeckia species for their major constituents.

We report the isolation and structure elucidation of two new

sesquiterpene lactones, the 12,6-trans-lactonized germacrolide

tamaulipin A angelate (4) and the eudesmanolide isogazaniolide (9).

Their structures were established by chemical and spectroscopic

methods. The molecular structures of tamaulipin A angelate (4) and

gazaniolide (8) were confirmed by single crystal X-ray diffraction.

Roots of R. nitida var. texana, roots and leaves of R.

scabrifolia, and aerial parts of R. maxima afforded the known

lignanes (+)-pinoresinol dimethyl ether (12) [12] and yangambin (13)

[13].

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. RESULTS AND DISCUSSION

The structures of all known sesquiterpene lactones and other

Rudbeckia constituents were established by spectral comparison with

published data (NMR, MS, IR).

An investigation of leaves of R. subtomentosa afforded the known

pseudoguaianolides 4-0-desacetyl1igulatine C (1) and ligulatine C (2)

[14]. Flower extracts provided the germacrolide costunolide (5) [15],

the eudesmanolides santamarine (10) [16] and reynosin (11) [17] as

well as the flavonol eupatolin (14) [18]. The roots yielded

costunolide (5), 11,13-dihydrocostunolide (6), gazaniolide (8) [19]

and the new lactones tamauplipin A angelate (4) and isogazaniolide

(9). The isolation of santamarine and reynosin is a strong indication

that these two lactones are artifacts formed during work-up

procedures by cyclization of 1(lO)-epoxycostunolide (7) [15,20].

However, lactone 7 was not detected in our study.

Tamaulipin A angelate (4), ^ crystalline compound

with NMR spectral signals (Table 1) typical for an

a-methylene-r-lactone moiety with two one-proton doublets at 36.28

(H-13b) and 5.54 (H-13a), both being coupled to a one-proton ddd at

32.58 (H-7). A strong IR absorption at 1767 cm ^corroborated the

presence of a y-lactone moiety. Further, IR bands at 1715, 1653 and

1665 cm”^ indicated the presence of an unsaturated ester and double

bond(s). The ester side chain was assigned to an angelate group on

the basis of diagnostic NMR signals: a one-proton quartet of

quartets at 36.08 (H-3’) and a pair of three-proton doublets of

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. bond(s). The ester side chain was assigned to an angelate group on

the basis of diagnostic NMR signals: a one-proton quartet of

quartets at 86.08 (H-3’) and a pair of three-proton doublets of

quartets at 81.99 (H-4’) and 1.83 (H-5’), together with strong mass

spectral peaks at m/z 83 [A^]* and 55 [A^]^. 2D COSY studies

suggested that H-7 was coupled to a signal at 84.54(dd, ^=9.7 Hz)

which was assigned to the lactonic proton at C-6. Two broadened

three-proton singlets at 81.60 and 1.79 were assigned to the methyl

group at C-10 and C-4, respectively. The position of the angelate at

C-2 of the medium ring skeleton was deduced from the chemical shift

(85.70, ddd) of the H-2 signal in the ^H NMR spectrum and 2D COSY

experiments. Two broad one-proton doublets at 84.98 and 4.99 were

assigned to H-1 and H-5 from the coupling patterns and their allylic

coupling to the C-lO-methyl and C-4-methyl, respectively. All other

proton signals were assigned on the basis of spin decoupling

experiments, 2D COSY and 2D ^H-^^C correlation (Table 1).

The ^^C NMR spectral signals (Table 2) were assigned by the use

of 2D COSY, ^H-^^C correlation and DEPT experiments. The coupling

constants between H-6 and H-7 (J^ ^=9.7 Hz) together with the allylic

coupling between H-7 and H-13 (J^ ^^=3.2 Hz) indicated a trans

attachment of the a-methylene-y-lactone ring [21]. Furthermore, the

CD spectrum of compound 4 exhibited a negative Cotton effect at 255

nm. This is in agreement with Geissman’s [22] and Beecham"s lactone

rules [23] for 12,6-trans-lactonic germacrolides, suggesting that the

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. new lactone represented the angelate derivative of tamaulipin A (3)

[24].

Single crystal X-ray diffraction analysis of tamaulipin A

angelate (4) verified the proposed structure, stereochemistry and

medium ring conformation. The molecular structures of one of the two

independent, molecules in the unit cell of crystalline compound 4 is

illustrated in Figure 1. The identity of the compound is confirmed to

be tamaulipin A angelate. The conformations of the two independent

molecules differ appreciably only at the angelate substituent, in

which the two differ by a torsional rotation of 176.6 deg. about the

C16-C17 bond. The ten-membered rings of both are in the

-conformation with crossed, trans double bonds [25]. The ring

conformations of the two molecules agree quite closely. The average

deviation calculated over 15 endocyclic torsion angles is 2.5.° The

maximum deviation is less than 5 degrees. The conformation of the

y-lactone ring can be described as a half-chair, with C12 lying on

the local pseudodiad. The conformation of the ring system compares

favorably with that of tamaulipin A [26]. The average deviation

between the 15 endocyclic torsion angles of tamaulipin A and that

from the average of the two molecules of tamaulipin A angelate is

only 3.6.°

Bond distances are normal. The bond lengths of the two molecules

in the crystal agree well, except in the angelate substituents, where

the conformation also differs.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. The main component of the root extract of R. subtomentosa was

gazaniolide (8) with a molecular formula The spectroscopic

data of 8 were identical with those previously reported [19].

Structure and stereochemistry of 8 were established by NMR, 2D

COSY and shift correlations. and DEPT allowed assignments

of all NMR signals which are included in Table 2. The structure

and stereochemistry of eudesmanolide 8 were confirmed by single

crystal X-ray diffraction. The molecular structure of gazaniolide is

illustrated in Figure 2.

Isogazaniolide (9), exhibited a NMR spectrum (Table

1) which was similar to that of gazaniolide (8). Besides the signals

typical of the a-methylene lactone group, the NMR spectrum of

compound 9 showed one lowfield doublet at 54.67 which was assigned

to H-6. 2D COSY and spin decoupling indicated that its vicinal proton

(H-7) was coupled with H-13 (J^ ^^^=3.4 Hz) and two further protons

(H-8a and H-8/3), consistent with the argument that the lowfield

doublet is due to H-6. Signals typical of an olefinic methyl at C-4

(51.90) and a quaternary methyl group at C-10 (51.16) were also

present. The coupling (9.7 Hz) indicated a trans-lactone. The

major difference between the ^H NMR data of eudesmanolide 9, when

a=7 129(1), b=l1.126(2), 0=1 6 .1 1 2 (2 ) A, V=1277.9(3)A^, Z=4, Dc=l.197 g cm , p(CuK )=5.8 cm , R=0.046 for 1298 observations with I>3o'(I). The coordinates are given in supplementary material, and the details of the structure determination will be published elsewhere.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 58

compared with those of compound 8, was the appearance in 9 of a two

broad doublets (J=21.5 Hz) centered at 62.67 and 2.58 which was

assigned to H-3a and H-3|3. Also, consistent with the presence of a

4,5-double bond was the absence of a signal for H-5. Spin-spin

decoupling indicated long range couplings between H-3 and H-15; H-3

and H-6 as well as H-3 and H-1. Long range couplings were also

observed between H-6 and H-15 and between H-9 and H-14. The NMR

spectrum of isogazaniolide is similar to that of gazaniolide except

for a triplet at 639.1 assigned to C-3 and a singlet at 6128.7 (C-5).

Chemical studies of three species of the subgenus Macrocline, R.

nitida var. texana, R. scabrifolia and R. triloba were also

performed. Extracts from the roots and aerial parts of these three

species afforded the known lignanes (+)-pinoresinol dimethyl ether

(12) [12,13], and yangambin (13) [13], but no sesquiterpene lactones

were detected.

The spectral data of (+)-pinoresinol dimethyl ether (12) agreed

with data reported in the literature [12,13] and the structure was

confirmed by single crystal X-ray diffraction studies [27]. The

structure of yangambin (13) was deduced from spectral comparison with

reported data [13]. Lignanes 12 and 13 are the first representatives

to be reported from the genus Rudbeckia.

It is of interest to note that in R. grandiflora [3] and R.

subtomentosa different parts of the plant contain different skeletal

types of sesquiterpene lactones. In both species, characteristic root

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 59

constituents are germacrolides and in R. subtomentosa also

eudesmanolides. In contrast, leaves contain pseudoguaianolides only.

In R. subtomentosa, flowers provide costunolide and eudesmanolides,

possibly being artifacts formed from the highly labile

constunolide-1(10)-epoxide. Floral parts in R. grandiflora [3] also

contain eudesmane esters. The ecological significance of the above

distribution patterns of sesquiterpene lactones within the plant is

not known.

EXPERIMENTAL

R. subtomentosa (Pursh) was collected on 20 September 1986 in

pinewoods and the roadside of US Hwy. 165 in Rapides Parish,

Louisiana, U.S.A., 0.5 miles north of the Allen Parish line south of

Greenmora Sec 1, TIS, R3W. (P. Cox and family; Cox No 4923, voucher

deposited at L.S.U., U. S. A.). Air-dried leaves (75 g) were ground

and extracted with CH^Cl^ according to the general procedure [28]

providing 2.0 g of the crude terpenoid extract, 1.7 g of which was

separated by CC on silica gel using hexane-EtOAc mixtures of

increasing polarity, 151 fractions of 25 ml each being collected.

Upon further prep. TLC of the various fractions, the lactones 1 and 2

were isolated. Fraction 76 provided 50 mg of 1 and fractions 89-91

gave 54 mg of 2. From flowers (165 g), 5.5 g of crude extract was

obtained. Part of the crude extract (4.4 g) was separated by CC on

silica gel using petrol-EtOAc mixtures of increasing polarity, 50

fractions of 25 ml each being collected. Fraction 16 provided 29 mg

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. of 10 and fractions 37-39 gave 15 mg of 11. A mixture of ^-sitosterol

and stigmasterol was isolated from fraction 20.

The air-dried root material (333 g) was ground and extracted with

petrol, CHCl^ and MeOH. After removing the solvent, the petrol

extract (8.0 g) was chromatographed over silica gel (55 g) using

petrol and petrol-EtOAc mixtures (95:5, 9:1, 4:1, 1:1) and finally

EtOAc. 21 fractions (250 ml) were obtained. Fraction 1 (1 g), after

rechromatography over silica gel and TLC purification, yielded

caryophyllene, caryophyllene oxide, p-selinene, a mixture of

glycerides, and a new sesquiterpene lactone, isogazaniolide (9).

Fraction 2-11 yielded mixtures of gazaniolide (8) [19] and

isogazaniolide (9). These less polar fractions provided gazaniolide

which was crystallized from petrol, mp. 75-76°C. Fraction 13 was

separated and purified by successive prep. TLC, yielding a

costunolide (5). Fractions 14-16 contained costunolide (5) and traces

of 11,13-dihydrocostunolide (6). Rechromatography of fractions 17 and

18 gave a mixture of glycerides, p-sitosterol, stigmasterol and

tamaulipin A angelate (4).

The CHCl^ extract (1.5 g) was chromatographed over silica gel (40

g) and eluted with petrol and mixtures of petrol-EtOAc (95:5, 9:1,

4:1) providing 12 fractions which contained lactones 4, 5, 8 and 9.

R. nitida var. texana (Perdue) was collected on 12 July 1987,

from roadside seep areas within open woods along Louisiana Hwy. 118,

east of Mora, in Natchitoches Parish (P. B. Cox; Cox No. 5211,

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 61

voucher deposited at L.S.U., U. S. A.). The air-dried roots (50 g)

were ground and extracted with CH^Cl^ providing 1.1 g of the crude

material 0.8 g of which was separated by CC on silica gel using

hexane-EtOAc mixtures of increasing polarity, 38 fractions of 25 ml

each being collected. Upon further prep. TLC of the various fractions

the lignanes 12 and 13 were isolated. Fractions 18-20 provided 35 mg

of compound 12 and fractions 35-38 afforded compound 13 (34 mg).

R. maxima (Nett) was collected on 7 August 1987 along roadside

and in pastures on Texas Hwy. 21, 3 miles east of Jet. Texas. 69 and

Alto, Texas, Cherokee Co. Texas (P. Cox, L. Urbatsch, A. W. Lievene;

Cox No. 5119; voucher deposited at L.S.U., U. S. A.)

The air-dried leaves (85 g) were ground and extracted with

CH^Cl^ according to the general procedure [28] providing 2.3 g of

crude terpenoid extract, 1.8 g of which was separated by CC on silica

gel using hexane-EtOAc mixtures of increasing polarity, 65 fractions

of 25 ml each being collected. Upon further prep. TLC of the various

fractions, the lignanes (12 and 13) were isolated. Fraction 36

provided 50 mg of 12 and fractions 43-51 gave 54 mg of 13.

R. scabrifolia (L. E. Brown) was collected on 12 July 1987 on

seepage area along the roadside and pastures of Plainview Comm. Rd. 2

miles North of Jet of US Hwy. 171, north of Hornbeck, Louisiana. (P.

Cox No. 5208; voucher deposited at L.S.U., U.S.A.)

The air-dried leaves and roots (55 g) were ground and extracted

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. of the crude terpenoid extract, 0.9 g of which were separated by CC

on silica gel using hexane-EtOAc mixtures of increasing polarity, 41

fractions of 25 ml each being collected. Fraction 26 provided 30 mg

of 12 and fractions 29-31 gave 44 mg of 13.

Rudbeckia grandiflora (Sweet) DC. was collected on 7 June 1986,

in the pinewoods and along the roadside of Louisiana Hwy. 121, 3.3

miles west of Rapides Parish line east of La Camp, sec 16, T 2N, R5W.

(P. Cox, L. Urbatsch and E. Harris Cox No. 4889, voucher deposited at

L.S.U., U.S.A. ).

Air-dried roots (50 g) were ground and extracted according to

the general procedure [28] providing 2.0 g of the crude terpenoid

extract 1.6 g of which was separated by CC on silica gel using

CHCl^-hexane mixtures of increasing polarity. Fractions (104) of 20

ml each, were collected, which upon crystallization by vapor

diffusion [29] from petrol-CHCl^, provided pure colorless crystals of

compound 4.

Tamauli

128-130°; UV A nm: 226; CD (MeOH; c 2.12 X lO"'*), [8]^^^ +2.1 x

10*. [0)255 -3.0 X 10^; IR cm"\ 1767 (y-lactone), 1715, 1653

(unsat ester), 1665 (double bonds); HRMS (FAB, probe) 70 eV, m/z

(rel. int.): 330.1857 [M]^ (79), (calc, for 330.1855),

230.1832 [M-A]* (35), 215.1800 [M-A-Me]* (2), 187.1642 [M-A-Me-CO]"

(10), 83.1617 [A^]* (100), 55.1227 [A^]'' (27).

X-Ray data for tamaulipin A angelate (4). A crystal of

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. dimensions 0.16x0.20x0.64 mm was used for data collection on an

Enraf-Nonius CAD-4 diffractometer equipped with CuK^ radiation

(X=l.54184 A) and a graphite monochromator. Crystal data are;

*"20^26*^4’ '"O'^oclinic space group P2^, a=15.757(1),

b=7.376(1) c=17.196(2)A, g=lll,25(l)°, V=1863(5) A^, Z=4, dc=1.178g

cm”^, p(CuK^)=6. 2 cm”^, T=26°C. Data were collected by w-28 scans of

variable speed, 0.92-3.30 deg min”^. One quadrant of data having

2°<0<7O°was measured, yielding 3814 unique data, of which 2713 had

I>3

corrections for background, Lorentz, polarization, and absorption

effects, the latter based on 'I' scans. The structure was solved by

direct methods using MULTAN [30], and refined by full-matrix least

squares using the Enraf-Nonius SDP programs [31]. Carbon and oxygen

atoms were refined anisotropically, while hydrogen atoms were located

from difference maps and included as fixed contributions. Convergence

was achieved with R=0.049, Rw=0.057 for 433 variables, and the

maximum residual electron density 0.55 eA“^. Final coordinates are

listed in Table 3; other data have been deposited with the Cambridge

Crystallographic Data Centre.

Acknowledgements-This research was supported by the Louisiana

Education Quality Support Fund (86-89)-RD-A-13 and the National

Science Foundation Biotechnology Program (Project No. EET-8713078).

Purchase of 100 and 400 MHz NMR spectrometers was made possible by

NIH Shared Instrumentation Grant 1 SIO RR0459-01.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. REFERENCES

1. Herz, W. and Kumar, N. (1981) J. Org. Chem. 46, 1356.

2. Jakupovic, J., Jia, Y., King, M. R. and Bohlmann, F. (1986) Liebigs Ann. Chem. 1474.

3. Vasquez, M., Macias, F. A., Urbatsch, L. E. and Fischer, N. H. (1988) Phytochemistry 27, 2198.

4. Waddell, T. G., Elkins, S. K. , Mabry, M. A., Singri, B. P., Wagner, H., Seligman, 0. and Herz, W. (1976) Indian J.Chem. 14B,

5. Jauhari, P. K. , Sharma, S. C., Tandon, J. S. and Dhar. M. M. (1979) Cent. Drug Res. Inst. 18 (2), 359.

6 . Atkinson, R. E. and Curtis, R. F. (1965) Tetrahedron Lett. 5,

7. Bohlmann, F. and Kleine, K. M. (1965) Chem. Ber. 98, 3081.

8 . Bohlmann, F., Grenz, M. , Wotschokowsky, M. and Berger, E. (1967) Chem. Ber. 100, 2518.

9. Herz, W. and Kulanthaiyel, P. (1985) Phytochemistry 24, 89.

10. Valadon, L. R. G. and Mummery, R. S. (1971) Phytochemistry 10,

11. Flores, H. , Hoy, M. W. and Pickard, J. J. (1987) Trends Biotechnol.5, 64.

12. Erdtman, H. (1936) Syensk Kem. Tid. 48, 236.

13. Pelter, A., Ward, R. S., Rao, E. V. and Sastry, K. V. (1976) Tetrahedron 32, 2783.

14. Maldonado, E., Mendoza, G. 0., Cardenas, J. and Ortega, A. (1985) Phytochemistry 24, 2981.

15. El-Feraly, F. S. and Chan, Y. M. (1978) J. Pharm. Sci. 67, 347.

16. Romo de Viyar, A., and Jimenez, H. (1965) Tetrahedron 21, 1741.

17. Yoshioka, H., RenoId, W., Fischer, N. H. , Higo, A., and Mabry, T. J. (1970) Phytochemistry 9, 823.

18. Quijano, L., Malanco, F. and Rios, T. (1970) Tetrahedron 26,

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 19. Bohlmann, F. and Zdero, C. (1979) Phytochemistry 18, 323.

20. Rodriguez, A. A., Garcia, M. and Rabi, J. A. (1978) Phytochemistry 17, 953.

21. Samek, Z. (1970) Tetrahedron Lett. 671.

22. Stôcklin, W., Waddell, T. G. and Geissman, T. A. (1970) Tetrahedron 26, 2397.

23. Beecham, A. F. (1972) Tetrahedron 28, 5543.

24. Fischer, N. H. , Mabry, T. J. and Kagan, H. B. (1968) Tetrahedron 24, 4091.

25. Fischer, N. H., Oliyier, E. J. and Fischer, H. D. (1979) in Progress in the Chemistry of Organic Natural Products (Herz, W., Grisebach, H. and Kirby, G. B. , eds). Springer, Vienna.

26. Witt, M. E. and Watkins, S. F. (1978) J. Chem. Soc. Perkin Trans II, 204.

27. Vasquez M., Fronczek, F. R. and Fischer, N. H. (1989) Acta Cryst. C45, 000.

28. Fischer, N. H., Wiley, R. A., Lin, H. N. , Karimian, K. and Politz, S. M. (1975) Phytochemistry 14, 2241.

29. Jones, P. G. (1981) Chem. Br. 17, 222.

30. Main, P. Hull, S. E., Lessinger, L., Germain, G., Declercq, J. P. and Woolfson, M. M. (1978). MULTAN, A System of Computer Programs for the Automatic Solution of Crystal Structures from X-Ray Diffraction Data. University of York (England) and Louvain (Belgium).

31. Frens, B. A. and Okaya, Y. (1980) Enraf-Nonius Structure Determination Package, Enraf-Nonius, Delft.4.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1. H NMR spectral data of compounds 4, and 9 (400 MHz.CDCl , IMS as internai standard)

1 4.98 d 5.38 dt 23 5.70 ddd 5.57 dt 3a 2.20 dd 2.67 AB q 33 2.73 dd 2.57 AB q 5 4.99 d -

6 2.58 ddd 2.63* 1.62 dddd’’’ 2.09 dq 83 1.62 dddd’*' 1.50-1.70'’' 9a 2.46 ddd 1.50-1.70'*' 93 2 . 16 ddd 1.50-1.70* 13a 5.54 d 5.45 d 13b 6.28 d 6.14 d 1.60 s 1.16 s 15 1.79 s 1.90 brs 3' 6.08 qq - 4' 1.99 dq - 5' 1.83 d -

Compound 4 J (Hz): l,2g=2g,1=9.6; 2p,3a=10.3; 20,33=5.4;3a.3^=8.27; 5,6=9.1; 6 ,7=9.7; 7,13a=3.2; 7, 133=3. 4. 3* , 4' =7. 1; 3', 5' =4' , 5’ =1. 5. Compound 9: 1,2=10.3; 1,3=2.0; 2,3a=2, 33=3. 3; 3a,33=21.5; 6,7=11.5; 7,8a=8a,93=8a, 93=3. 0; 7,83=83,9a=11.7; 7,13a=3.1; 7,13b=3.4; 8a,83=12.8 ; 83,93=4.4.

Obscured by other signals. ^Overlapped with H-3a signal.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. c NMR data of compounds 4, 8 and 9 (50.32 MHz.CDCl , IMS as internai standard)

c 4 8 9

1 126. 79d'’’ 120.2 * 120.7

2 71.05d 136.9 135.3

3 45.23t 1 22.7d 39.lt 4 193.00s 135.9 123.6^ 5 50.7 128.7^s 6 81.52d 81. 1 83.0 7 50.79d 50.7 50.0 8 27.93t 21.3 22.9 9 40.92t 36.5 34.6 10 140.99s 37.4 38.4

11 138.8 139.2 12 2 0 1 .00s 170.5 170.4 13 119.92t 116.5 118. 1 14 17.15q 15.3 15 18.44q 21.9 1’ 162.23s - -

2 ’ - - 3' 138.Old - - 4' 15.83q - - 5' 20.57q - -

Assignments were confirmed by C- H chemical shift correlation. ^Peak multiplicity was obtained by heteronuclear multipulse programs. ^Multiplicities are not repeated if identical with those in the preceding column. ^Interchangeable signals.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Positional parameters and their esd’s compound Atom X Y z Beg iA^I

GIA 0.3959(2) 0.000 0.4778(2) 5.85(8) 02A 0.4413(2) 0.0510(6) 0.6144(2) 7.9(1) 0.0870(2) -0.2996(5) 0. 1140(2) 5.45(7) 0.1302(2) -0.2875(6) 0.0041(2) 7.2(1) 0.1844(2) -0.4105(7) 0.2465(2) 4.7(1) 0.1821(2) -0.3098(7) 0.1701(2) 4.8(1) 0.2165(3) -0.1126(7) 0.1906(2) 5.2(1) 0.3052(2) -0 . 1088(6) 0.2658(2) 4.23(9) 0.2997(2) -0.1043(6) 0.3403(2) 4.44(9) 0.3709(2) -0.1585(6) 0.4214(2) 4.5(1) C7A 0.3360(3) -0.3007(7) 0.4693(2) 4.8(1)

C8A 0.3416(3) -0.4998(7) 0.4482(2) 5.4(1) C9A 0.2583(3) -0.5785(7) 0.3773(2) 5.2(1) ClOA 0.2510(2) -0.5220(6) 0.2907(2) 4.26(9) CllA 0.3900(3) -0.2454(7) 0.5578(2) 5.5(1) C12A 0.4132(3) -0.0515(8) 0.5568(2) 5.6(1) C13A 0.4150(3) -0.3420(9) 0.6265(3) 7.8(2) C14A 0.3234(3) -0.5942(7) 0.2614(3) 5.9(1) C15A 0.3897(3) -0.1368(8) 0.2475(2) 5.5(1) C16A 0.0713(3) -0.2877(7) 0.0325(2) 4.8(1) C17A -0.0287(3) -0.2696(7) -0.0163(3) 5.2(1) C18A -0.0613(3) -0.2558(7) -0.0978(3) 6 .2 (1 ) C19A -0.0926(3) -0.2667(9) 0.0342(3) 6.9(1) C20A -0.0134(3) -0.2611(9) -0.1569(3) 7.0(1)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 3. Positional parameters and their esd’s compound 4 (cont.) Atom X Y z Beg (A^)

OIB 0.4879(2) -0.2637(5) 0.8994(2) 5.78(8) 02B 0.6374(2) -0.3149(6) 0.9521(2) 7.9(1) 03B 0. 1115(2) 0.0596(5) 0.5960(2) 6.28(8) 04B -0.0175(2) 0.0399(7) 0.6217(2) 8.9(1) CIB 0.2507(3) 0.1658(7) 0.6992(2) 4.90 C2B 0. 1645(3) 0.0687(7) 0.6861(2) 5.4(1) C3B 0. 1842(3) -0.1314(8) 0.7169(3) 5.9(1) C4B 0.2581(2) -0.1351(6) 0.8021(2) 4.6(1) C5B 0.3433(3) -0.1481(6) 0.8058(2) 4.7(1)

C6B 0.4272(2) -0.1035(7) 0.8790(2) 4.6(1) C7B 0.4848(2) 0.0446(6) 0.8588(2) 4. 15(9)

C8B 0.4574(3) 0.2421(7) 0.8670(2) 5.0(1) C9B 0.3905(3) 0.3284(7) 0.7847(2) 5.3(1) 0.2917(3) 0.2754(6) 0.7634(2) 4.6(1) 0.5788(2) -0.0109(7) 0.9147(2) 4.9(1) C12B 0.5756(3) -0.2093(7) 0.9257(2) 5.5(1) C13B 0.6538(3) 0.0864(9) 0.9534(2) 6.5(1) C14B 0.2489(3) 0.3501(8) 0.8201(3) 6.5(1) C15B 0.2284(3) -0.1052(9) 0.8744(3) 6.3(1) C16B 0.0201(3) 0.0426(7) 0.5740(3) 6 .1 (1 ) C17B -0.0336(3) 0.0268(8) 0.4804(3) 7.2(1) 0.0030(3) 0 .0 2 1 1 (8 ) 0.4305(3) 7.4(1) C19B -0.1412(3) 0.0175(9) 0.4576(3) 7.3(2) C20B 0.0934(3) 0 .0 2 2 1 (1 ) 0.4288(3) 8 .0 (2 )

The equivalent isotropic thermal parameter, for atoms refined anisotropically, is defined by the equation: ;[a\, + h \ . c B + abB cosg + acB cosb + bcB cosa]

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 3, R'OH X'CHj 4, R = OAng X'CHg 5, R'H X'CHg 6 , R'H X'H.ocCHj 7, R'H, 1(10) X'CHj epoxide

Û ""'CZ/ Meo ^

HO-)-C-rÇ=CH-M ! I Me 100; 83 : 55

Fig. 2.1. Compounds isolated from R. grandiflora, R.subtomentosa, R. nitida var. texana and R. maxima

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ppm

Fig. 2, 100 MHz H NMR spectrum of the crude extract of leaves of R. subtomentosa in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. p p m 7 6 5 4 3 2 1

Fig. 2.3. 100 MHz NMR spectrum of the crude extract of aerial parts of R. scabrifolia in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. iî

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 2.6. 100 MHz H NMR spectrum of the crude extract of roots of R. subtomentosa in deuteriochloroform. Spectrum provided by Dr. Leovigildo Quijano.

M 8 . 9 5 - ü l g 0 . 9 0 -

I 0 . 8 5 - M h

0.75 0.729 4000 3500 3000 2500 2 1 5 0 0 WAVEHUMBER Fig. 2.8. Infrared spectra of tamaulipin A angelate (2.4) (film < KBr window). Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. AngO/,

6.0 5.0 4.0 3.0 2.0 1.0 PPM

angelate (2.4). Chemical shifts are given in ppm relative to Me Si (solvent C^D^)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 2.11 2D J-Resolved spectrum of tamaulipin A angelate (2.4) in

Fig. 2.13. Proton broadband decoupled (BB) and DEFT spectra of tamaulipin A angelate (2.4) at 50.3 MHz in deuteriochloroform.

- 20 20,0 300

Fig. 2.15. Circular Dichroism spectrum of tamaulipin A angelate (2.4) in methanol.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 13b I3a

Fig. 2.18. 400 MHz ( H, H) COSY-45 spectrum of Isogazaniolide (2.9). Chemical shifts are given in ppm relative to He Si. Sample provided by Dr. Leovigildo Quijano.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ^ O I!

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 7

D E P T 90 “

DEPT 135“

140 100 60 20 PPM

Fig. 2.20. DEPT spectra of isogazaniolide (2.9) at 50.3 MHz in deuteriochloroform. Sample provided by Dr. Leovigildo Quijano.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CHAPTER 3

SESQUITERPENE LACTONES FROM RUDBECKIA MOLLIS

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. (to be submitted to Phytochemistry)

Marta Vasquez, Leovigildo Quijano , Lowell E. Urbatsch

and Nikolaus H. Fischer*

Louisiana State University, Baton Rouge, LA. 70803, U.S.A.

(Received )

K ey Word Index-Rudbeckia mollis-, Asteraceae; Heliantheae; polyacetylenes; sesquiterpene lactones; pseudoguaianolides; eudesmanolides; flavonol.

ABSTRACT-Four new pseudoguaianolide type sesquiterpene lactones, llocH,13-dihydrorudmollin, 15-acetyl-lloH,13-dihydrorudmollin 4-acetyl-llaH,13-dihydrorudmollin and 15-acetylconfertin were isolated from Rudbeckia mollis. In addition, the known compounds rudmollin, rudmollin diacetate, 15-acetylrudmollin, 4-acetylrudmollin, thiarubrine B, thiophene B, isoalantolactone endodiplophyllolide, p-selinene, trans-trans-farnesol, squalene and eupatolin, are reported. Their structures including the stereo chemistry were determined from spectroscopic data and chemical transformations.

INTRODUCTION

The genus Rudbeckia, of the tribe Heliantheae (Asteraceae), six

species of which are found in the southeastern United States, is

characterized by perennial or annual herbs and shrubs, nearly all of

which exhibit morphological variability. Thus far, ten species of

Rudbeckia including the ornamental summer-flowering annual R. mollis

Permanent address: Institute de Quimica, Universidad Nacional Autonoma de México. México 20, D.F. * Author to whom correspondence should be addressed.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. [1] have been investigated chemically [1, 2, 3]. The aerial parts of

Rudbeckia mollis from Florida were previously investigated and

provided C-15-funtionalized ambrosanolides[1]. Our collection from

another Florida location gave the previously described compounds,

plus a series of other known and new natural products.

In the present study, aerial parts gave four new structurally

related pseudoguaianolides,another new lactone was prepared from a

known lactone.Also, four known pseudoguaianolides [1] and the known

flavonol glycoside eupatolin [4] were obtained. Roots provided the

known terpenoids g-selinene, trans,trans-farnesol, and squalene. In

addition, the acetylenes thiarubrine B (10) and thiophene B (11), the

eudesmanolides isoalantolactone (12), endodiplophyllolide (13) and

8-epi-diplophyllolide (14) were isolated. The structures of the known

and new compounds were elucidated by spectroscopic methods and

chemical transformations.

RESULTS AND DISCUSSION

The extract of aerial parts of R. mollis afforded, after column

chromatography and further purification by preparative TLC, the known

lactones rudmollin (1), diacetylrudmollin (3), 15-acetylrudmollin (5)

and 4-acetylrudmollin (7). In addition, two new compounds,

llaH,13-dihydrorudmollin (2) and llocH,13-dihydro-15-acetylrudmollin

(6) were found. The structures of the known lactones (1,3,5,7 and 9)

were determined by the use of NMR and NMR spectra and direct

comparison of their physical properties with those of previously

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. reported data [1]. The structure of the new compounds were determined

on the basis of their and NMR spectral data (Tables 1, 2 and

3) as well as mass spectral fragmentation patterns, chemical

transformations and IR, UV, and CD spectral studies.

llocH,13-Dihydrorudmollin (2) was obtained as a mixture with rudmollin

(1), and the two lactones could not be completely separated. However,

inspection of the NMR data (Table 1) of pure compounds 1 and 2

clearly indicated that both compounds had to be present in the

mixture isolated from R. mollis. This was verified by reduction of

the mixture of 1 and 2 with NaBH^in methanol which provided pure

llodl, 13-dihydrorudmollin (2). Upon treatment of the mixture of

lactones 1 and 2 with acetic anhydride/pyridine a mixture of acetates

3 and 4 resulted.

15-Acetoxyconfertin (9) was previously reported as an oxidation

derivative of 1 [1] but to our best knowledge this is the first

report of its natural occurrence.

llaH,13-Dihydrorudmollin (2),

absorption at 3420 cm ^ and NMR signals (Table 1) similar to those

of compound 1, the structure of which has been elucidated by X-ray

analysis [5]. In lactone 2, the signals of the exomethylene protons

were not present. Instead, a three-proton doublet at 51.20 (C-11

methyl) appeared. Upon irradiation of the C-11 methyl doublet at

51.20 (J=7.4 Hz), a doublet of a quartet at 52.95 (IH, ^^=7.4 and

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 94

sesquiterpene-y-lactone [6]. The p-configuration of the C-10 methyl

followed from the NOE between H-10 and H-8 and the absence of a NOE

between H-14 and H-1 indicated a |3 configuration at C-1. The lactonic

proton (H-8) appeared at 64.63 (ddd). The coupling constant

(J^ ^=11.4 Hz) indicated a cis-diaxial disposition of H-7 and H-8

[7]. The H-4 proton gave a doublet of doublets at 63.87 (J^ ^^=4.0,

^^=7.8 Hz) suggesting a p-orientation of the 4-hydroxyl group [8].

The signals corresponding to H-15 appear as broad two-proton AB

doublets at 64.15 and 3.74 ppm and a carbon triplet at 65.89 ppm,

typical of a CH^OH group attached to quaternary carbon. These

signals, as well as all the remaining signals of the ^H NMR spectrum,

were assigned by spin decoupling, 2D COSY, spin decoupling and NOE

difference experiments (Table 1).Furthermore, in compound 2 the

paramagnetic acylation shift of the doublet of doublets at 63.92

(H-4) to 65.5 in 4 (A6=1.58) was consistent with H-4 being geminal to

the hydroxyl group. In situ acylation of lactone 2 with

trichloroacetylisocyanate [9] yielded the trichloroacetylcarbamate

derivative 2a, the ^H NMR spectrum of which showed two NH signals at

68.55 and 8.78 ppm, providing further evidence for the presence of

two hydroxyl groups in compound 2. The presence of a secondary

hydroxyl group at C-4 was again suggested by the paramagnetic

acylation shift of H-4 from 63.87 in compound 2 to 64.70 ppm in

compound 2a (A6=0.88). Also, the shift of the respective H-15 from

64.15 and 3.74 in compound 2 to 64.6 and 64.25 ppm in compound 2a

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. (AS=0.45, AS=0.49) provided further evidence for the presence of

hydroxyl groups at C-4 and C-15.

The NMR signal, corresponding to C-8, was identified by

single frequency resonance decoupling and heteronuclear multipulse

experiments (DEPT), as a doublet at 80.74 ppm; C-6 as a triplet at

26.13 and C-7 as a doublet at 338.43 ppm. The NMR spectral

assignments of compound 2 are summarized in table 3. The data

resemble the spectra of lactone 1 except for the signals due to C-13,

a quartet at 310.50 ppm and C-11, a doublet at 329.15 ppm.

Compound 4, showed in its IR spectrum strong

absorptions at 1740 and 1250 cm ^(acetate). In the NMR spectrum of

compound 4, several signals were nearly identical to those in the

spectrum of compound 3 [1]. These signals were a ddd at 4.60 ppm (H-8

g=ll.4 Hz), a dddd at 32.66 (H-7), a doublet at 30.92 (H-14), a

double of doublets at 34.78 (H-4), a two proton signal at 34.08

(H-15a, H15b), and finally, two singlets at 32.04 and 2.00 (OAc)

indicating the presence of two acetate groups in the molecule.

However a doublet at 31.07 ppm was present which was assigned to

H-13. These signals, together with the absence of doublets at 36.31

and 5.70 ppm, suggested that compound 4 was the

llctH,13-dihydroderivative of 3. Furthermore, a doublet quartet at

32.85 (H-11) was present. 2D COSY studies suggested that H-7 was

coupled to a signal at 34.60 (ddd ^=11.4 Hz).This signal was

assigned to the lactonic proton at C-8. A three-proton doublet at

30.92 was assigned to the methyl group at C-10. The position of the

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 96

acetate at C-4 was deduced from the chemical shift (34.78 dd) of the

H-4 signal in the NMR spectrum and 2D COSY experiments. All other

proton signals were assigned on the basis of spin decoupling

experiments, 2D COSY and 2D ^H-^^C correlation (Table 1).

On the basis of the similarities of the NMR spectral data of

compounds 3 and 4, the stereochemical structure of the

pseudoguaianolide skeleton of compound 4 is suspected to be the same

as in compound 3. The coupling constants between H-7 and H-8

(J^ g=l1.4 Hz) indicated the cis nature of the y-lactone ring.

Further evidence was provided by the CD spectrum of compound 4 which

exhibited a positive Cotton effect at 238 nm. This effect is as

predicted for 12,8-cis-lactonic pseudoguaianolides by Geissman’s [10]

and Beecham’s lactone rule [11]. The ^^C NMR spectral signals (Table

3) were assigned by the use of ^H-^^C correlation and DEPT

experiments. The ^^C NMR of compound 4 (Table 3) supported the

presence of three carbonyl groups, and four methyl groups. The

multiplicity of the signals in the ^^C NMR spectrum indicated that

compound 4 bears an oxygen substituent at C-4. In order to verify

the proposed structure and stereochemistry, single crystal X-ray

diffraction data of lloH,13-dihydrorudmollin diacetate (4) were

obtained [1].

Compound 5: The ^H NMR spectrum of compound 5 was in part

similar to that of compound 3. However a changed situation at C-4

followed from the differences of the H-4 signals which was shifted

upfield in the spectrum of compound 5. X-ray diffraction analysis of

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. compound 5 confirmed the structure assigned by Herz [1]. Compounds 5

and 6 were obtained as a mixture. They could not be separated from

each other. Reduction of the mixture gave compound 6.

llaH, 13-Dihydrorudmollin 15-0-acetate (6), A strong

IR absorption band at 1770 cm”^ corroborated the presence of a

y-lactone. Further IR absorption bands at 1741 and 1248 cm~^

indicated the presence of an acetate. This compound exhibited

structural features similar to those of sesquiterpene lactone 5.

Compound 6 displayed a one-proton doublet of quartets at 52.90

(H-11), two three-proton doublets at 61.15 (H-13, ^^=7.4 Hz) and

60.92 (H-14), and a three-proton singlet at 62.00 (OAc). The signal

of the lactonic proton appeared at 64.51 (H-8 ^=11.4 Hz) coupled

with a signal at 62.66 (H-7). A signal at 62.90 ppm in turn was also

coupled to a methyl group, which was responsible for a doublet at

61.15 (H-13 ^^=7.4 Hz). As for the stereochemistry, the

cis-fusion of the lactonic ring, and the p-orientation of H-8 could

be deduced from the values of the coupling constants (Table 1). The

NOE difference spectrum confirmed that H-7 and H-8 were cis, that

H-10 was cis to H-8, and that H-8 was cis to H-11. NOE was observed

between H-7 and H-15 while no effect was observed between H-15 and

H-4.

The above findings established the complete relative

stereochemistry shown in formula 6. The ^^C NMR spectral signals

(Table 3) were assigned on the basis of 2D COSY, ^H-^^C correlation

and DEPT experiments.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Compound 7: The NMR spectrum of compound 7 [1] showed that

this lactone differed from lactone 5 in the nature of the oxygen

group at C-4 and C-15.

Compound 8,

function at C-15 was suggested by the mass spectrum (loss of 42

units). In part, the NMR signals were similar to those of compound

7 [1]. However, a doublet at 51.18 H-13, a signal at 52.90 H-11 and

the absence of doublets at 56.25 and 5.60 ppm indicate that compound

8 is the llocH, 13-dihydroderivative of 7. From the ^^C NMR spectrum

the presence of three methyls (C-13 10.53, C-14 16.76 and OAc 21.24

ppm), two carbonyls and three carbons bonded to oxygens (C-4 78.53,

C-15 65.09 and C-8 80.16 ppm) were also suggested. Spin decoupling of

the ^H NMR spectrum allowed the assignment of all signals. NOE

difference spectroscopy further elucidated the stereochemistry and

the structure deduced from the decoupling experiments. The structure

of llaH,13-dihydrorudmollin 15-OAc was verified by single crystal

X-ray diffraction.

Hexane and CH^C

separation and TLC purification, the bioactive polyacetylenes

thiarubrine B (10) and thiophene B (11) [12]. Other root constituents

included p-selinene, trans-trans-farnesol, squalene, a mixture of the

known eudesmanolide, isoalantolactone (12) and its 4,5-endo-isomer

(13). A mixture of compounds 12 and 13 has been recently isolated

from Eupatorium quadrangularae and showed ant-repellent activity [13,

14]. The NMR spectral data of compound 13 were identical with those

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 99

of diplophyllolide A [15], a eudesmanolide which was previously

isolated from the liverwort Diplophyllum albicans [15] and which

belongs to the enantiomeric group of lactones [16]. Assuming that

higher plants produce without exception sesquiterpene lactones with

7(xH-configuration (7S), compound 14 and the one isolated from E.

quadrangularae [14] should be representedent-diplophyllolide A (13)

[15]. This was supported by a negative Cotton effect at 255 nm caused

by the n — > n transition of the methyllene lactone chromophore in

the CD spectrum [11] of the mixture. This is opposite to the Cotton

effect of diplophyllolide A [15]. We propose the name

enantio-diplophyllolide for compound 13. Structure (13) was recently

also assigned to a compound named isoalloalantolactone, which had

been isolated from Inula racemosa [17]. Inspection of the published

NMR data, particularly the allylic coupling constant between H-7

and H-13 (J^ ^^=3 Hz) indicated that the structure of the compound

named isoalloalantolactone [17] should be revised to stereostructure

14, which is the 8-epimer of compound 13.

EXPERIMENTAL

Plant material. Rudbeckia mollis (Ell) was collected on 1 July

1988, in the pinewoods and along the roadside of the intersection of

Florida State Route 20 and Istanbul Road, 4.0 miles east of the

Junction of State Route 20 and southbound Putman County Road 21, West

of Interlachen, Florida. (P. Cox, L. Urbatsch and A. Lievens No. Cox

5072; voucher deposited at L.S.U., U.S.A.).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 100

The air-dried floral parts (50 g) were ground and extracted with

CH^Cl^ providing 2.0 g of the crude terpenoid extract. The crude

extract of the aerial parts (1.6 g) was separated by CC on silica gel

using hexane-CHCl^ mixtures of increasing polarity. 14 fractions of

200 ml each being collected. Upon further prep. TLC of the various

fractions, the sesquiterpene lactones 1-9 were isolated. Fraction 7

provided 60 mg of colorless compound 1 upon crystallization by vapor

diffusion from petrol-CH^Cl^ [18]. Fractions 8-9 afforded a mixture

of compounds 1 and 2 (28 mg), fractions 10-11 gave a mixture of

compounds 5 and 6, fractions 12-13 yielded compound 7 (38 mg) and

fraction 14 gave compound 9.

The air-dried root material (162 g) was ground and extracted with

hexane and then CH^Cl^ providding 1.5 and 1.0 g of crude extract

respectively. The crude hexane extract (1.5 g) was separated by CC on

silica gel (30 g) using hexane and mixtures of hexane-EtOAc of

increasing polarity, 11 fractions of 150 ml each collected. Fraction

1, when eluted with hexane, provided p-selinene and fraction 2 gave

further amounts of g-selinene, thiophene B (11) and squalene. From

fraction 3 thiarubrine B (10) was isolated. Further CC of fraction 9

afforded a mixture of ^-sitosterol and stigmasterol and a mixture of

isoalantolactone (12) and its 3,4-endo isomer enantio-diplophyllolide

(14) and trans-trans-farnesol.

+2.1x10 ; IR y cm” : 1767 (y-lactone); LRMS; m/z (rel. int.) 268.2

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 101

[M]" (79), 250.2 (35), 232.2 [M-2H^0]* (2), 217.2

[M-2H^0-Me]* (10), 189.2 [M-2H^0-Me-C0] (10), 83.2 (100). H NMR see

Table 1; see Table 3.

Acétylation of compound 2. A sample of compound 2 (53 mg) was

treated with acetic anhydride (2 ml) and pyridine (0.1 ml) at 25»C

for 1.5 hr. and worked up under standard conditions. Purification by

CC (CH^Cl^-acetone) yielded 10 mg of diacetate (4).

llccH, 13-Dihydrorudmollin diacetate C H 0 IR 1768 cm"^ 19 28 6 max (y-lactone), 1740 and 1250 (acetate); UV (MeOH) nm: 242; CD

(MeOH; c 2.02x10'“): [0]^^^ +3. lxlo“; IR cm'\ LRMS; m/z (rel.

(17), 190.3, 95 and 81.

gum; UV nm: 243; CD MeOH; c 2.05xl0“'‘): [0]^^^ +2.9x10^; IR

*"m!x • 3420, 1770, 1741, 1248 (OAc). LRMS ; m/z (rel. int.)

310.3 [M]" (0.9), 290.3 [M-HOAc]* (1.2), 232.7 [M-RCO^H]'" (60), 220.0

[M-RCO^H-Me]* (159), 181.0 [M-RCO^H-Me-CQ]* (17), 141.0 [RCO]" (20).

Reduction of compound 3. A sample of compound 3 (5 mg) was

treated with NaBH^ in methanolic solution at room temp, for 20 min.

After acidification, CH^Cl^ extraction and prep. TLC, 2 mg of

compound 4, were obtained.

llaH,13-dihydrorudmollin 4-0-acetate (8). (C^^H^^O^) colorless

gum; UV nm: 242; CD (MeOH; c 2.00x10'“): [0]^^^ +2.9xlo“; IR

v™'": cm'“: 3421, 1772, 1740, 1247 (OAc). LRMS ; m/z (rel. int.)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 102

310.4 [M]* (1.9), 290.4 [M-HOAcl* (1.4), 230.6 [M-RCO^H]* (61), 220.0

[M-RCOH-Me]* (148), 180.9 [M-RCOH-Me-CO]* (17),140.9 [RCO]^ (19).

Acknowledgements-Ve thank Dr. Tirso Rios, Institute de Quimica,

Universidad Autonoma de México for a sample of eupatolin. This

research was supported by the Louisiana Education Quality Support

Fund (86-89)-RD-A-13 and the National Science Foundation

Biotechnology Program (Project No. EET-8713078). Purchase of 100 and

400 MHz NMR spectrometers was made possible by NIH Shared

Instrumentation Grant 1 SIO RR0459-01.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. REFERENCES

1. Herz, W. and Kumar, N. (1981) J. Org. Chem. 46, 1356.

2. Bohlmann, F. Jakupovic, J. and Zdero, C. (1978) Phytochemistry 17, 2034.

3. Vasquez, M., Macias, F. A., Urbatsch, L. E. and Fischer, N. H. (1988) Phytochemistry 27, 2195.

4. Quijano, L., Malanco, F. and Rios, T., (1970) Tetrahedron 26, 2851.

5. Vasquez, M., Fronczek, F., Quijano, L. and Fischer, N. H. (1989) Acta Cryst. submitted.

6. Narayanan, C. H. and Venkatasubramanian, N. K. (1968) J. Org. Chem. 33, 3156.

7. Fischer, N. H., Oliyier, E. J. and Fischer, H. D. (1979) in Progress in the Chemistry of Organic Natural Products (Herz, W. , Grisebach, H. and Kirby, G. B., eds. ) Springer, Vienna, pp.

8. Bohlmann, F. and Zdero, C. (1982) Phytochemistry 21, 647.

9. Samek, K. and Budesinsky, M. (1979) Collect. Czech. Chem. Comm. 44, 558.

10. Stocklin, W., Waddell, I. G. and Geissman, T. A. (1970) Tetrahedron 26, 2397.

11. Beecham, A. F. (1972) Tetrahedron 28, 5543.

12. Towers, G. H. N. (1987) Plant Physiol. 6, 85.

13. Okunade, A. L. and Wiemer, D. F. (1985) Phytochemistry 24, 1199.

14. Hubert, T. D., Okunade, A. L. and Wiemer, D. F. (1987) Phytochemistry 6, 1751

15. Benesoya, V., Samek, Z. and Vasickoya, S. (1975) Collect. Czech. Chem. Comm. 40 1967.

16. Asakawa, Y. (1982) in Progress in the Chemistry of Organic Natural Products (Herz, W., Grisebach, H. and Kirby, G. B., eds. ) Springer, Vienna, pp.

17. Kaur, B. and Kalsi, P. S. (1985) Phytochemistry 24, 2007.

18. Jones, P. G. (1981) Chem. Br. 17, 222.

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Page(s) m issing in num ber only; text follows. Film ed as received.

UMI

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 105 NMR spectral data of compounds 2, 3, 4, 6 and 8 (200 MHz, TMS as internal standard) 6 2 4 8

H CDCI3 CgDe* CDCI3 CDCI3 CDCI3

1 1.50-1.54* 1.01 ddd 1.50-1. 5# 1.55-1.4^ 1.65 1.54 dddd° 1.48-1.5f 1.46-1.,5^ 1.09-1.5# 1.51 dddd 2P 1.76 dddd 1.76 1.74 1. 10 1.76-1.92* 2.14 ddd 1.53 1.52 1.55-1.46* 1.76-1.92* 33 1.80 ddd 2.05 2. 18 1.75 2.20 dddd 3.87 dd 4.04 4.78 3.76 4.85 dd 1.96 dd 1.80 1.29-1.,40* 1.50 1.65 d 63 1.62 dd 1.42 1.52-1.,78* 1.68 1.47 dd 7 2.66 dddd 2.61 2.66 dddd 2.63 2.65 ddd 8 4.63 ddd 4.78 4.60 4.68 4.63 dddd 9a 2.20 ddd 1.78 2. 10 2. 18 2.18 ddd 93 1.68 ddd 1.72 1.87 1.72 1.75 dddd 10 1.90-2.06* 0.67 dq 1.94-2.,07* 1.95-2.07* 1.15-2.02* 113 2.95 dq 2.50 2.50 2.85 2.95 1.20 d 6.21 1.07 1.28 1.20 133 - 5.24 d - - - 14 0.93 d 0.67 0.92 0.98 0.99 15 4.15 d 4.02 4.08 4.08 4. 15 15’ 3.74 d 3.90 3.96 3.92 3.75 OAc - 1.65 s 2.02 2.07 2.06 1.64s -

The signals due to hydroxyl groups are omitted. Multiplicity and coupling constants are the same as reported by Herz et al [1]. ^Obscured ^Multiplicities are not repeated if identical with those in the previous column.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2. Coupling Constants for Compounds 2, 3, 4, 6 and 8 in Hertz

H.H 2 1.10 6.7 5.8 1,2a -— 3.6 8.9 — 1.2p -— 8.7 7.5 — 2p,2a 12. 1 12. 1 11.6 11.7 12. 1 2a, 3a 9. 0 8.9 8.0 2.6 11.8 2p,3a 2.0 1.9 2.0 2.0 12.6 2p,3p ——- 4.7 - 3a, 4 8.8 6.9 3a, 3p 11.9 12.6 10.0 6a, 6p 12.6 14.6 14.6 6a, 7 2.4 2.6 2.6 5.6 3.6 6p,7 5.4 9.2 2. 4 3. 1 7,11 9.8 9.2 9.2 9.2 7,8 11.4 7. 1 11.4 11.9 8,9a 10.0 9.5 6. 4 6.9 8,9p 2.8 2.3 6.3 6.3 9a, 9g 10.8 12.6 12.6 12.6 12.4 9a, 10 1.7 2.7 -- 10.4 9p,10 10.4 10.4 10.4 9. 1 — 10, 14 6.4 6.4 6.9 6.4 11, 13 7.3 7.3 7.3 7.4 15a,15b 11.9 11.9 11.3 11.3

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 3. C NMR data of compounds 2, 3, 4, 6 and 8 (50.32 MHz, CDCl^, TMS as internal standard)

c 2 3 4 6

1 42.41d'"' 44.06* 43. 15 45.48

2 22.53t 23.25 22.98 22.30 22.69

3 29.59t 26.83 24.48 29.70 25.25

4 80.74d - 77.55 79.98 82.29 80. 16

5 47.71s 47.28 46.61 53.52 46.67

6 26.13t 34. 18 35.95 90.09 36.29

7 38.43d 36.97 38.04 43.44 38.27

8 80.39d 78.87 77.99 83.91 78.53

9 36.46t 29.89 26.43 24.80 29.19

10 37.38d 35.88 37.29d 32.76 37.09

11 29.ISd 140.55s 29.00d 29.88 29.69

12 178.78s 168.82 178.40 170.23 179.45

13 lO.Slq 122.lit 10.28q 10.89 10.53

17.42q 15.87 16.36 16.95 16.76

65.90t 64.02 63.74 61.82 65.09

170.45s 170.94 170.87 170.82

169.79s 170.66 21.24

20.60q 21.01

20.60q 21.02

Assignments for compound 3 were confirmed by C - H chemical ^ shift correlation. Peak multiplicity was obtained by heteronuclear multipulse + programs. Multiplicities are not repeated if identical with those in preceding column.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 3.1. Compounds isolated from Rudbeckia mollis.

A* OR, x" R R. 1 H H 2 H /S-CHj.H 2o TAC 3 Ac CH, 4 Ac Ac /8-CH,.H 5 H Ac OH, 6 H Ac 7 Ac H CH, 8 Ac H

H A

CH»{c h c ]j^^C =C CH = CHj CH,-{c = c jj^ ^ C H C C H = CH^

Çp^“ Vo

TAC=-C-NH-COCCIj

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Page(s) missing in num ber only; text follows. Film ed as received.

UMI

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Fig. 3.2 100 MHz fMÎ spectrum of the crude extract of leaves of Rudbeckia mollis in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ppm 100 MHz NMR spectrum of the crude extract of flowers of Rudbeckia mollis in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ppm 7 6 5 4 3 2 1

Fig. 3.4. 100 MHz NMR spectrum of the crude extract of aerial parts of Rudbeckia maxima in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. g*-S

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. A 111. M h À A W

® % »

4.5 4 0 3.5 3.0 2.5 2.0 1.5 1.0

dihydrorudmollln (3.2). Chemical shifts are given in ppm relative to Me Si.

Fig. 3.8. Proton broadband decoupled (BB) and DEFT spectra of llctH, 13-dihydrorudmolIin (3.2) at 50.3 MHz in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 3.9. 400 MHz NMR spectrum of lloH,13-dihydrorudmollin (3.2) after in situ acylation with TAI.

70 60 5.0 40 3.0 2.0

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. (ndeg) 10

OH OH 0

10 200

Fig. 3.10. Circular Dichroism spectrum of llaH,13-dihydrorudmollin (3.2) in methanol.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. % = 1 ill

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. OAc

AcO

45 4.0 3.5 3.0 2.5 2:0 Fl.g. 3.13. 400 MHz {^H. ^H) COSY-45 spectrum of 11 ocH, 13- dihydrorudmollin diacetate (3.4). Chemical shifts are given in ppm relative to Me^Si.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. -20.0

-10.0

- 0.0

- 10.0

- 20.0

4 5 4.0 3.5 2.5 2.0 1.5 PPM

Fig. 3.14 2D J-Resolved spectrum of llaH,13-dihydrorudmollin diacetate (3.4).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. o

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. kW,kiV ALMi

80 70 60 50 40 30 20 10 p

Fig. 3.16. Proton broadband decoupled (BB) and DEFT spectra of llaH,13-dihydrorudmollin diacetate (3.4) at 50.3 MHz in deuteriochloroform.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. W e g )

AcO

200

Fig. 3.17. Circular Dichroism spectrum of lloH,13-dihydrorudmollin diacetate (3.4) in methanol.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Dc/itirlniail of Clwmisin/ [LSffl Louisiana State U niversity a BATON ROUGE • LOUISIANA • 71)f«3-lS(M

Executive Secretary International Union of Crystallography 5 Abbey Square Chester CHI 2IIU England

As the primary author of the article "Structure of the iignane (+)-pinoresinol dimethyl ether" aceptcd to bo published in Acta Crystallographica. 1989, COO, 0000.

Marta Vasquez / Box E-4, Choppin Hall Department of Chemistry Louisiana State University Baton Rouge, LA 70803

I / " // -

y' \ Cj L^'-’ ^ \

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CHAPTER 4

STRUCTURE OF THE LIGNANE (+)-PINORESINOL DIMETHYL ETHER

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 127

(Reprinted with permission from Acta Crystallographica Section C,

Crystallography)

V=2004.5(10) fr, Z=4, 0=1.280 g cm'=". CuK^, X=l. 54184 A, p=7.2

cm"\ F (000)=824, 1=298»K, R=0.038 for 1733 observations (of 2350

unique data). The compound was isolated from Rudbeckia maxima

Nutt, R. nitida Perdue and R. scabrifolia Brown (Asteraceae) which

were collected in Alto, Texas; East Baton Rouge Parish, Louisiana

and in Vernon Parish, Louisiana, respectively. The two

five-membered rings of the central dioxabicyclooctane system are

cis-fused, each ring adopting the half-chair conformation with one

atom lying on both pseudodiads. The phenyl rings are planar within

maximum deviation 0.Oil(3)A, and the four methoxy substituents lie

near these planes, with CCOC torsion angle magnitudes ranging

1.7(4)-6.5(4)°

Experimental. Pinoresinol dimethyl ether (1) was obtained as

colorless needles, dimensions 0.15x0.28x0.45 mm mounted in a

capillary,due to failure of epoxy glue to harden properly in

presence of this compounds. Space group from absences hOO with h

odd, OkO with k odd, 001 with 1 odd. Enraf-Nonius CAD-4

diffractometer with graphite monochromator, cell dimensions from

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 128

setting angles of 25 reflections having 24>0>19.° Data collection

by w-20 scans designed for I=50o'(I), subject to max. scan time =

120s. Scan rates varied 0.53-3.30°mln"^. Reflections having

4 <20<15O , 0

background, Lorentz-polarlzatlon, and absorption by ^ scans,

minimum relative transmission 0.9596, 2350 unique data, no

redundant data. Standard reflections 200, 020, 002, ±2.3% maximum

random variation, no decay correction. Structure solved using

MULTAN 78 (Main, Hull, Lesslnger, Germain, Declercq & Woolfson,

1978) refinement by full-matrlx least squares based on F with

weights w=4Fo^[

I>2.5o'(I), (617 unobserved reflections), using Enraf-Nonius SOP

(Frenz & Okaya, 1980). Non-H atoms anisotropic; H atoms located by

F and Included as fixed contributions. Isotropic B’s were assigned

to H atoms, equal to 1.3 Beq of the bonded carbon atoms. Positions

of methyl H atoms were adjusted using AF maps, while other H atoms

positions were calculated with C-H 0.95Â. Atomic scattering

factors of Cromer & Waber (1974) and anomalous coefficients of

Cromer (1974). Final R=0.03806 (0.066 all data), wR=0.04105,

S=l.989 for 254 variables, extinction coefficient g=l.57(10) 10

where the correction factor (1+gIc)”^ was applied to Fc, max.

shift In final cycle 0.03o-, max. residual density 0.13, mln.

-0. 14e A Refinement of the enantlomorphous structure yielded

R=0. 03820,wR=0.04116, S=l.994. Atomic coordinates and equivalent

Isotropic thermal parameters for the former refinement are given

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 129

in table 1* , bond distances and angles in table 2, and endocyclic

torsion angles describing the conformation in table 3. Fig. 2

shows the atom-numbering scheme.

Related Literature Description of lignanes (Haworth, 1936);

synthesis of (+), (-) and (±) forms of pinoresinol dimethyl ether,

(Erdtman, 1936); reaction with bromine to yield dibromo

derivatives (Erdtman, 1935); characterization of lignanes having a

2, 6-diaryl-cis-3,7-dioxobicyclo[3.3.0]-octane structure (Adjangba,

1963 and Hearon, 1955); unit cell and space group determination of

dibromo and diiodo derivatives of pinoresinol dimethyl ether,

establishing twofold molecular symmetry (Wang Lund, 1960); crystal

structure determination of the lignane (-)-syringaresinol (Bryan,

1976), crystal structure determination of the lignane

(-)-3,6-bis-(3,4-dimethoxyphenyl)tetrahydro-lH,3H-furo-[3,4c]furan

-1,4-diol (Ghisalberti, 1987).

* Table of H atom parameters, anisotropic thermal parameters, and structure factors have been deposited with the British Library Document Supply Centre as Supplementary Publication No. Sup#### (##pp.). Copies may be obtained through the Executive Secretary, International Union of Crystallography, 5 Abbey Square, Chester CHI 2HU, England.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Adjangba, M. S., (1963) Bull. Soc. Chim. France, 2344-2358.

Bryan, R. F.& Fallon, L. (1976) J. C.S. Perkin I, 341-345.

Cromer, D. T. (1974). International Tables for X-ray

Crystallography,Vol. IV, Table 2.3.1. Birmingham: Kynoch Press.

(Present distributor Kluwer Academic Publishers, Dordrecht.)

Cromer, D. T.& Waber, J. T. (1974). International Tables for

X-ray Crystallography, Vol. IV, Table 2.2.B. Birmingham: Kynoch

Press.(Present distributor Kluwer Academic Publishers,

Dordrecht.).

Erdtman, H., (1935) Ann. 513, 229-239.

Erdtman, H., (1936) Svensk Kem. Tidskr. 48, 236.

Frenz, B. A.& Okaya, Y.(1980) Enraf-Nonius Structure Determination

Package. Delft, Holland.

Ghisalberti, E. L., Jefferies, P. R. , Skelton, B. W. & White, A.

H., (1987) Aust. J. Chem. 40, 405-411.

Haworth, R. D., (1936) Ann. Rep. Prog. Chem. 33, 266.

Hearon, W. M.& MacGregor, W. S., (1955) Chem. Rev. 55, 957-1068.

Main, P. Hull, S. E., Lesslnger, L. , Germain, G., Declerq, J.&

Woolfson, M. M. (1978) MULTAN 78 A system of Computer Programs for

the Automatic Solution of Crystal Structures from X-ray

Diffraction Data.University of New York, England and Lovain,

Belgium.

Wang Lund, E., (1960) Acta. Chem. Scand. 14 No.2, 496-497.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1. Positional parameters and their esd’s of (+)-pinoresinol dimethyl ether Atom X y z B^q(A^)

CÏ 0.2462(3) 0.7726(2) 0.4361(1) 3.82(6) C2 0.3640(3) 0.7382(2) 0.4913(1) 3.76(6) C3 0.4005(3) 0.6302(2) 0.4694(1) 4.89(5) C4 0.4037(4) 0.6324(3) 0.3884(2) 5.19(8) C5 0.2882(3) 0.7139(2) 0.3625(1) 3.90(6) 0.3454(4) 0.8041(2) 0.3098(1) 4.04(6) 0.3585(3) 0.8985(2) 0.3567(1) 5.24(5) C8 0.2508(4) 0.8920(2) 0.4147(2) 5.11(7) C9 0.3259(3) 0.7416(2) 0.5748(1) 3.49(5) CIO 0.2745(3) 0.6499(2) 0.6120(1) 3.68(6) 0.2360(3) 0.6545(2) 0.6883(1) 3.54(6) 0.2469(3) 0.7533(2) 0.7275(1) 3.69(6) 0.2975(3) 0.8434(2) 0.6910(2) 4.29(7) C14 0.3383(4) 0.8372(2) 0.6149(2) 4.46(7) 015 0.1872(3) 0.5680(2) 0.72985(9) 4.58(5) C16 0.1678(5) 0.4679(2) 0.6911(2) 6.02(9) 0.2027(2) 0.7496(2) 0.80229(9) 4.49(5) 0.2267(5) 0.8466(3) 0.8458(2) 6.27(9) C19 0.2570(4) 0.8271(2) 0.2389(1) 3.70(6) C20 0.3294(3) 0.8392(2) 0.1701(1) 3.73(6) C21 0.2542(4) 0.8612(2) 0.1039(1) 3.94(6) C22 0.1047(3) 0.8746(2) 0.1064(1) 4.02(6) C23 0.0332(3) 0.8632(3) 0.1746(2) 4.63(7) C24 0.1096(4) 0.8387(3) 0.2401(2) 4.78(7) 025 0.3158(3) 0.8732(2) 0.0332(1) 5.69(5) C26 0.4676(4) 0.8653(3) 0.0281(2) 5.97(9) 027 0.0398(3) 0.9000(2) 0.0385(1) 5. 18(5) C28 -0.1112(4) 0.9196(4) 0.0398(2) 7.2(1) The equivalent isotropic thermal parameter, for atoms refined anisotropically, is defined by the equation: ^[a B + b B + c B + abB cosg + acB cosb + bcB cosa]

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ! 2: Bond Distances (A) and Angles ( ) of (+)-pinoresinol dimethyl ether

Cl C2 1.519(3) Cll C12 1.401(3) Cl C5 1.534(3) Cll 015 1.368(2) Cl C8 1.518(3) C12 C13 1.365(3) C2 03 1.425(3) C12 017 1.380(2) C2 C9 1.513(3) C13 C14 1.393(3) 03 C4 1.426(2) 015 C16 1.421(3) C4 C5 1.534(3) 017 C18 1.436(3) C5 C6 1.541(3) C19 C20 1.392(3) C6 07 1.431(3) C19 C24 1.371(4) C19 1.519(3) C20 C21 1.384(3) C8 1.428(3) C21 C22 1.393(3) C9 1.389(3) C21 025 1.376(2) C9 C14 1.377(3) C22 C23 1.377(3) CIO Cll 1.391(3) C22 027 1.373(2) C23 C24 1.386(3) 027 C28 1.418(4) 025 C26 1.410(3)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2: Bond distances (A) and Angles of (°) (+)-pinoresinol dimethyl ether cont.

103.1(2) C8 107.9(2) 114.1(2) Cl C8 07 104.6(2) C5 Cl C8 103.8(2) C2 C9 CIO 121.0(2) Cl C2 03 104.9(2) C2 C9 C14 120.2(2) Cl C2 C9 116.5(2) CIO C9 C14 118.8(2) 03 C2 C9 110.1(2) C9 CIO Cll 120.6(2) C2 03 C4 104.9(2) CIO Cll C12 119.5(2) 03 C4 C5 107.1(2) CIO Cll 015 124.7(2) Cl C5 C4 103.5(2) C12 Cll 015 115.8(2) Cl C5 C6 104.9(2) Cll C12 C13 119.9(2) C4 C5 C6 114.3(2) C13 C12 017 114.8(2) C5 C6 07 105.5(2) C12 C12 017 125.3(2) C5 C19 116.4(2) 120.1(2)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 3: Selected Torsion Angles ( ) of (+)-pinoresinol dimethyl

C5 Cl C2 03 -33.3 C5 Cl C2 C9 -155.4 C8 Cl C2 03 -145.2 C8 Cl C2 C9 92.7 C2 Cl C5 C4 12.7 C2 Cl C5 C6 -107.4 C8 Cl C5 C4 132.0 C8 Cl C5 C6 11.9 C2 Cl C8 07 81.5 C5 Cl C8 07 -30.0 Cl C2 03 C4 41.9 C9 C2 03 C4 168.0 C2 03 C4 C5 -33.4 03 C4 C5 Cl 11.7 03 C4 C5 C6 125.2 Cl C5 C6 07 10. 1 Cl C5 C6 C19 -114.3 C4 C5 C6 07 -102.6 C5 C6 07 C8 -30.2 C19 C6 07 C8 97. 1 C5 C6 C19 C20 -134.8 C6 07 C8 Cl 38.3 CIO Cll 015 C16 -3.3 Cll C12 017 C18 173.8 C22 C21 025 C26 177.2 C21 C22 027 C28 -176.9

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 4.1. Single crystal X-ray structure of (+)-pinoresinol dimethyl ether (4.1).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CHAPTER 5

STRUCTURES OF THREE SESQUITERPENE y-LACTONES

FROM RUDBECKIA HOLLIS

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 137

(submitted to Acta Crystallographica C)

by Marta Vasquez, Frank R. Fronczek, Leovigildo Quijano

and Nikolaus H. Fischer

Department of Chemistry, Louisiana State University,

Baton Rouge, Louisiana 70803, U S A

(Received )

P2^2^2^, a=9.0071(10), b=10.053(2), c=18.144(2) A, V=1642.9(7) A^,

Z=4, Dx=1.247 g cm"^, A(MoK^)=0.71073 A, p=0.85 cm"\ F (000)=664,

T=295°K, R=0.049 for 1540 observations having I>3o-(I) (of 2158 unique

data). (2): Rudmollin diacetate, Mr=350.4, monoclinic, P2^,

a=8.117(2), b=7.681(4), c=14.865(3) A, p=92.47(2)°, V=926.0(6) A^,

Z=2, Dx= 1.257 g cm"^, A(CuK^)=l. 54184A, fx=7.29 cm"\ F (000) =376,

T=295°K, R=0.044 for 3170 observations having I>3

data). (3): Dihydrorudmollin, Mr=268.4, orthorhombic,

P2^2^2^, a=9.441(2), b=10.731(2), c=13.349(2) A, V=1352.3(7) A^, Z=4,

Dx=1.320 g cm'^, A(MoK^)=0.71073 A, p=0.88 cm“\ F (000)=584, T=125°K,

R=0.043 for 2293 observations having I>3

Crystals of the three compounds were isolated from Rudbeckia mollis

Ell, (Asteraceae) which was collected in Putman County Florida. All

three compounds are ambrosanolide-class pseudoguaiano1ides. The

dihydro compound (3) has its methyl group C13 g-oriented. All

compounds have the seven-membered rings in twist-boat conformations

with CIO on the pseudodiad, and the cyclopentane ring in envelope

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 138

conformations with C5 at the flap. Hydroxyl groups of both compounds

bearing them are involved in hydrogen bonds.

Introduction. Recently, we have identified a new sesquiterpene

lactone (3) which was isolated from R^ mollis (Vasquez et ai.). From

the same plant we obtained two other sesquiterpene lactones (1, 2),

previously reported by Herz, Kumar, and Blount (1981). The chemical

structures of compounds 1, 2 and 3 were established on the basis of

of and NMR spectroscopic studies and chemical transformations.

The molecular structures of pseudoguaianolides 1, 2 and 3 were

determined in order to confirm the relative configurations at all

chiral centers of these molecules.

Experimental. Intensity data for all three compounds were obtained

from fragments of colorless needles, on Enraf-Nonius CAD-4

diffractometers equipped with either MoK^ (X=0.71073 A) or CuK^

(A=l.54184 A) radiation and graphite monochromators. Data collection

parameters are summarized in Table 1. The cryogenic (125 K) data for

(3) were collected using a gas stream cryostat. Variable scan

rates were employed in the w-28 scans, and a maximum was set on the

time spent on a weak reflection. Cell dimensions were obtained from

the setting angles of 25 reflections, including measurements at ±20.

Intensity standards were remeasured every 10,000 seconds, and linear

decay corrections were applied for (2) and (3). One octant of data

was collected within the specified 0 limits for (1) and (3), while a

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. full sphere of data was collected for (2). Data reduction included

corrections for background, Lorentz, and polarization effects.

Absorption corrections for (2) were based on \j) scans.

Space groups, for (1) and (3) were uniquely determined from

systematic absences hOO with h odd, OkO with k odd, and 001 with 1

odd. The space group of (2) was determined from systematic absences

OkO with k odd and the known chirality of the compound. The

structures of (1) and (2) were solved by direct methods, using

MULTAN80 (Main, Fiske, Hull, Lesslnger, Germain, Declercq, &

Woolfson, 1980). compound (3) is isomorphous with rudmollin (Herz,

Kumar, & Blount, 1981), and coordinates for all nonhydrogen atoms

except C13 of rudmollin were used as a beginning refinement model.

Structures were refined by ful1-matrix least squares based on F

with weights w=4Fo^[

(Frenz & Okaya, 1980), scattering factors of Cromer & Waber (1974),

anomalous coefficients for (2) of Cromer (1974),and data having

I>3o'(I). Non-hydrogen atoms were refined anisotropically, and

hydrogen atoms were located by AF syntheses. For (1), H atoms were

included as fixed contributions with C-H distance 0. 95Â and B=1.3Beq

for the bonded atom. For (2), acetate H atoms were treated as above,

while other H atoms were refined isotropically. For (3), all H atoms

were refined isotropically. Secondary extinction coefficients g, were

refined for all structures, with the correction factor (1+gI )“^

applied to Fc. Final values of the extinction coefficients, residual

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 140

densities in final AF maps, R factors and other details of the

refinements are given in Table 1.

Atomic parameters for nonhydrogen atoms are given in Table 2-4,

the molecular structures are shown in Figures 1-3, bond distances are

given in Table 5, bond angles in Table 6, and selected torsion angles

in Table 7.

Discussion. All three compounds are shown to be pseudoguaianolides of

the ambrosanolide class, in which the methyl group C14 is /3. Methyl

group CIS of compound (3) is also shown to be oriented g. In all

compounds, the lactone ring is cis-fused to the seven-membered ring,

and the other five-membered ring is trans-fused. In all three

compounds, the seven-membered ring is in a twist boat conformation

with the pseudodiad passing through CIO and bisecting the C6-C7 bond.

The asymmetry parameter AC^ (Duax & Norton, 1975) has a value if 5.9°

for (1), 8.9° for (2), and 5.0° for (3). In all three structures, the

cyclopentane ring has the envelope conformation with C5 at the flap.

Asymmetry parameters AC are 2.6° for (1), 5.7° for (2), and 5.7° for

(3). The lactone ring is much flatter, and has the envelope

conformation with C7 at the flap for compounds (1) and (3), which

have asymmetry parameters AC 1.9° and 2.8°, respectively, while

Table of H atom parameters, anisotropic thermal parameters, and structure factors have been deposited with the British Library Document Supply Centre as Supplementary Publication No. Sup ( pp. ). Copies may be obtained through the Executive Secretary, International Union of Crystallography, 5 Abbey Square, Chester CHI 2HU, England.P

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. diacetate (2) has its lactone ring in the half-chair conformation,

with C12 on the pseudodiad, and 0.8°. The conformations of these

three pseudoguaianolides are analogous to that of rudmollin (Herz et

al. 1981). Rudmollin also has both five-membered rings in envelope

conformations with C5 and C7 at the flaps. Its seven-membered ring is

an intermediate conformation, and has been described by Herz et al.

(1981) as a boat, with pseudo mirror passing through Cl. Our

computations show that AC for that description is 14.3°, while AC^

for the twist boat with CIO on the pseudodiad (the conformation of

(1), (2), and (3)) is smaller, at 9.2°. Bond distances for our

compounds are normal, and show good agreement. Bond distances in

dihydrorudmollin (3) agree extremely well with those of rudmollin.

The 18 bond lengths not involving Cll, in which the hybridization

state differs, exhibit a rms deviation of O.OOSA, with the largest

individual difference 0.013 A for C8-C9. Both our compounds

containing OH groups exhibit hydrogen bonding in the solid. The

hydroxyl group 01 of acetate (1) forms an intermolecular hydrogen

bond with lactone carbonyl oxygen 03 at x, y+1, z, having 0— 0

distance 2.746(3)A. The angle about the unrefined H position is 173°.

The hydroxyl compound (3) exhibits both intramolecular and

intermolecular H bonding. The intramolecular bond involves 04 as

donor and hydroxyl group 01 as acceptor, 0— 0 distance 2.747(2)A,

angle at H 143(3)°. The intermolecular hydrogen bond involves 01 as

donor and hydroxyl group 04 at 1/2+x, 1/2-y, 2-z, with 0— 0 distance

2.841(2) A and angle at H 157(3)°.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 142

This research was supported by the Louisiana Education Quality

Support Fund (86-89)-RD-A-13 and the National Science Foundation

Biotechnology Program (Project No. EET-8713078). The purchase of the

diffractometer was made possible by an NSF instrumentation grant

(CHE-8500781).

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. REFERENCES

Bryan, R. F. & Fallon, L. (1976) JL Perkin 341-345.

Cromer, D. T. (1974). International Tables for X-ray

Crystallography, Vol. IV, Table 2.3.1. Birmingham: Kynoch Press.

(Present distributor Kluwer Academic, Publishers Dordrecht.)

Cromer,D. T.& Waber, J. T. (1974). International Tables for X-ray

Crystallography, Vol. IV, Table 2.2.B. Birmingham: Kynoch Press.

(Present distributor Kluwer Academic Publishers, Dordrecht. )

Duax, W. L. & Norton, D. A. (1975). Atlas of Steroid Structure.

Vol. I. New York: Plenum.

Frenz, B. A.& Okaya, Y.(1980) Enraf-Nonius Structure Determination

Package. Delft, Holland.

Herz, W. , Kumar, N. & Blount, J. F. Org. Chem. (1981) 46,

1356-1361.

Main, P. , Fiske, S. J., Hull, S. E. , Lesslnger, L., Germain, G.,

Declercq, J.-P.,& Woolfson, M. M. (1980) MULTAN 80 A System of

Computer Programs for the Automatic Solution of Crystal

Structures from X-ray Diffraction Data. Universities of York,

England and Louvain, Belgium.

Vasquez, M., Quijano, L., Urbatsch, L. , & Fischer, N. H.,

Phvtochemistrv. submitted.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1. Summary of data collection and structure refinement parameters 1 2 3 0. 33 x0 . 40 x0 .5 8 0. 33x 0. 38 x0 . 53 0.3 5x 0. 40 x 0. 52Crystal size (mm) 0.33x0.40x0.58 0. 33x0.38x0.53 0.35x0.40x0.52Crystal Radiation MoK CuK MoK Reflections used for cell constants e range (°) 10-13 25-30 w Scan width (°) 0.70+0.35tan0 0. 70+0.14tan0 0.80+0.35tan9 Scan speeds 0.56-4.00 1.03-3.33 0.71-4.00 (deg. min" ) Max.scan time per reflection (s) Range for data collection e ( ° ) 1-27.5 2-75 1-35 0.11 - 10,10 0,15 0,13 -9,9 0,17 0,23 -18,18 0,21 Standard Reflections 200,020,006 300,020,002 300,060,002 Crystal decay (%) 14.9 5.9 Empirical absorption correction max.trans. coefficient 0. 9960 min.trans. coefficient 0.8802 Reflections measured 2255 7695 3492 unique 2158 3747 3335 observed (I>3o-(D) 1540 3170 2293 0.020 R, wR 0.049, 0.056 0.044, 0.055 0.043, 0.043 Variables 200 306 269

Max. shift/e. s. d. ratio (A/o-) <0.01 0. 09 <0.01 Min./max.height in final AF (eA ) 0.19/0.19 -0.27/0.26 -0.24/0.39 Goodness of fit. 2.404 1.930 1.439 Extinction, g 5.0(9)xl0“‘’ 6.5(6)xl0"^ 3. I(10)xl0''^

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Standard. Deviations Compound 1 01 0.0478(3) 0.3790(2) -0.0041(1) 5.52(6) 02 0.1494(4} -0.2410(2) 0.0696(1) 6.33(7) 03 0.0799(5) -0.3536(2) -0.0302(2) 9.1(1) 04 -0.1666(2) 0.1378(2) 0.1398(1) 3.89(4) 05 -0.3337(3) 0.2859(3) 0.1782(2) 6.75(7) Cl 0.1968(3) 0.1670(3) 0.1419(2) 3.57(6) C2 0.2375(4) 0.3044(3) 0. 1717(2) 4.60(8) C3 0.2087(4) 0.4006(3) 0. 1072(2) 4.70(8) C4 0.1466(4) 0.3135(3) 0.0451(2) 3.90(7) C5 0.0730(3) 0.1938(3) 0.0839(2) 3.18(6) C6 0.0443(4) 0.0800(3) 0.0290(2) 3.61(6) C7 0.1724(4) -0.0199(3) 0.0217(2) 3.97(7) C8 0.1991(4) -0.1073(3) 0.0906(2) 4.34(7) 0.1206(4) -0.0721(3) 0. 1611(2) 4.24(7) CIO 0.1753(4) 0.0538(3) 0. 1984(2) 3.99(7) Cll 0.1430(6) -0.1219(3) -0.0381(2) 5.93(9) C12 0.1210(5) -0.2501(4) -0.0019(2) 5.78(9) C13 0.0973(6) -0.1020(3) -0.1071(2) 5.8(1) C14 0.0812(4) 0.0871(4) 0.2654(2) 4.93(8) CIS -0.0724(4) 0.2456(3) 0. 1165(2) 3.62(6) C16 -0.2971(4) 0.1724(3) 0. 1702(2) 4.25(7) C17 -0.3832(4) 0.0542(4) 0. 1915(2) 5.59(9)

The equivalent isotropic thermal parameter, for atoms refined

anisotropically, is defined by the equation:

I ",

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. I 3. Positional Parameters and Their Estimated Standard Deviations Compound 2

Atom X y z 01 0.2655(2) 0.000 0.5458(1) 8.05(5) 02 0.1013(3) -0.2321(4) 0.5452(1) 10.19(6) 03 0.4870(2) 0.2108(2) 0.83888(7) 4.56(3) 04 0.5483(2) 0.2050(3) 0.98566(9) 6.82(4) 05 0.0686(2) 0.4835(2) 0.87979(9) 5.27(3) 06 -0.1389(3) 0.6641(4) 0.8481(2) 10.98(6) 0.3447(3) 0.4884(3) 0.6922(1) 4.97(4) 0.3787(3) 0.6680(4) 0.7324(2) 6.44(6) C3 0.2358(3) 0.6983(4) 0.7960(2) 6.25(5) C4 0. 1406(3) 0.5270(3) 0.7943(1) 4.85(4) C5 0.2645(2) 0.3850(3) 0.7694(1) 4.02(4) C6 0.1778(2) 0.2139(3) 0.7413(1) 4.00(3) C7 0.1235(2) 0.1981(3) 0.6410(1) 4.56(4) C8 0.2690(3) 0.1829(4) 0.5769(1) 5.83(5) C9 0.4432(3) 0.2158(4) 0.6141(1) 5.59(5) CIO 0.4816(3) 0.4061(4) 0.6378(1) 5.95(5) Cll 0.0310(3) 0.0337(4) 0.6229(1) 5.54(5) C12 0.1302(3) -0.0810(4) 0.5675(2) 7.20(6) C13 -0.1102(4) -0.0213(5) 0.6552(2) 7.39(7) C14 0.6565(3) 0.4234(5) 0.6779(2) 7.13(7) CIS 0.3761(2) 0.3544(3) 0.8535(1) 4.29(4) C16 0.5658(2) 0.1474(3) 0.9118(1) 4.28(4) 0.6724(3) -0.0031(4) 0.8905(2) 6.46(5) CIS -0.0713(3) 0.5629(4) 0.8982(2) 6.53(5) C19 -0.1281(3) 0.5117(5) 0.9885(2) 7.95(7) The equivalent isotropic thermal parameter, for atoms refined

anisotropically, is defined by the equation: "Ii E, *,•

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ; 4. Positional Parameters and Their 1

Standard Deviations Compound 3 Atom X y z 01 0.7149(2) 0.1303(1) 1.0131(1) 1.42(2) 02 0.7896(2} 0.4811(1) 0.6248(1) 1.80(3) 03 0.8462(2) 0.6670(1) 0.6881(1) 2.39(3) 04 0.4785(2) 0.2531(1) 0.9449(1) 1.55(2) Cl 0.7116(2) 0.1082(2) 0.7389(1) 0.86(3) C2 0.6852(2) -0.0280(2) 0.7703(1) 1.14(3) C3 0.7214(2) -0.0318(2) 0.8826(2) 1.23(3) C4 0.7533(2) 0.1048(2) 0.9115(1) 1.02(3) C5 0.6725(2) 0.1844(2) 0.8335(1) 0.82(2) C6 0.7251(2) 0.3213(2) 0.8338(1) 0.98(3) C7 0.8490(2) 0.3466(2) 0.7635(1) 1.02(3) C8 0.8046(2) 0.3490(2) 0.6513(1) 1.20(3) C9 0.6650(2) 0.2889(2) 0.6218(2) 1.25(3) CIO 0.6576(2) 0.1474(2) 0.6347(1) 1.22(3) Cll 0.9175(2) 0.4763(2) 0.7753(2) 1.29(3) C12 0.8491(2) 0.5550(2) 0.6952(2) 1.61(3) C13 0.9157(3) 0.5381(2) 0.8775(2) 1.87(4) C14 0.5102(2) 0.1008(2) 0.6067(2) 1.79(4) CIS 0.5131(2) 0.1830(2) 0.8566(2) 1.15(3)

The equivalent isotropic thermal parameter, for atoms refined

anisotropically, is defined by the equation:

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 5. Bond Distances in Angstroms Compound 1

Atom 1 AtomJ Distance Atom 1 Atom 2 Distance

01 C4 1.422(4) C4 C5 1.544(4) 02 C8 1.467(4) C5 C6 1.539(4) C12 1.325(5) C5 CIS 1.529(4) 03 C12 1.218(5) C6 C7 1.535(4) 04 CIS 1.439(4) C7 C8 1.548(5) 04 C16 1.344(4) C7 Cll 1.516(5) 05 C16 1.196(4) C8 C9 1.504(5) Cl C2 1.528(4) C9 CIO 1.518(5) Cl CS 1.557(4) CIO 1.519(5) Cl CIO 1.543(4) Cll 1.460(5) C2 C3 1.541(5) Cll 1.333(5) C3 1.533(5) C16 1.471(5)

Atom 1 Atom 2 Distance Atom 1 Atom 2 Distance

01 C8 1.479(3) C4 1.540(3) 01 C12 1.314(3) C5 1.540(3) 02 C12 1.227(4) C5 CIS 1.530(2) 03 CIS 1.446(3) C6 C7 1.540(2) 03 C16 1.327(2) C7 C8 1.554(3) 04 C16 1.198(2) C7 Cll 1.488(4) 05 C4 1.460(2) C8 C9 1.517(3) 05 C18 1.328(3) C9 CIO 1.532(4) 06 C18 1.194(4) CIO C14 1.522(3) Cl C2 1.524(4) Cll C12 1.470(4) Cl C5 1.561(3) Cll C13 1.330(4) Cl 1.538(3) C16 C17 1.486(3) C2 C3 1.545(4) C18 C19 1.490(4) C3 C4 1.525(4)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 5. Bond Distances in Angstroms Compound 3 (cont.)

Atom 1 AtomZ Distance Atom 1 Atom 2 Distance

01 C4 1.430(2) CS C6 1.SSK2) 02 C8 1.468(2) CS CIS 1.535(3) 02 C12 1.3S2(3) C6 C7 1.524(3) 03 C12 1.207(3) C7 C8 1.556(3) 04 CIS 1.437(2) C7 Cll 1.542(3) Cl C2 1.541(3) C8 C9 1.519(3) Cl CS 1.549(3) C9 CIO 1.530(3) Cl CIO 1.540(3) CIO C14 1.526(3) C2 C3 1.538(3) Cll C12 1.508(3) C3 C4 1.545(3) Cll C13 1.517(3) CS 1.548(3)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 6. Bond Angles in Degrees Compound 1

Angle Angle tllLl tllLl tllLl tllLl======C8 02 C12 112.1(3) C8 C7 Cll 102.8(2) CIS 04 C16 116.2(2) 02 C8 C7 10S.2(3) C2 Cl CS 104.7(2) 02 C8 C9 107.0(3) C2 Cl CIO 117.S(3) C7 C8 C9 118.7(3) CS Cl CIO 119.1(2) C8 C9 CIO 115.0(3) Cl C2 C3 10S.0(3) Cl CIO C9 111.1(3) C2 C3 C4 10S.2(2) Cl CIO C14 116.0(3) 01 C4 C3 115.1(2) CIO C14 111.0(3) 01 C4 CS 112.3(3) C7 107.4(3) C3 C4 CS 10S.4(2) C7 128.6(3) Cl CS C4 97.8(2) C12 120.9(3) Cl CS C6 lis.4(2) 03 122.0(4) Cl CS CIS 114.3(2) Cll 110.7(3) C4 CS C6 110.8(2) Cll 127.2(4) C4 CS 106.2(2) 111.2(2) C6 CS 111.1(3) 04 122.5(3) CS C6 C7 114.6(3) 04 C16 111.1(3) C6 C7 C8 114.7(3) OS C16 126.4(3) C6 C7 Cll 111.9(3)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 6. Bond Angles in Degrees Compound 2 (cont.)

2 Angle Angle

C8 01 C12 112.2(2) 01 C8 C7 104.9(2) CIS 03 CIS lis.0 (1) 01 C8 C9 106.1(2) C4 OS C18 116.9(2) C7 C8 C9 119.0(1) C2 Cl CS 104.3(2) C9 CIO 114.8(2) C2 Cl CIO 117.1(2) CIO C9 111.6(2) CS Cl CIO 120.8(2) CIO C14 116.2(2) Cl C2 C3 104.4(2) C9 CIO C14 110.5(2) C2 C3 C4 104.7(2) C7 Cll C12 109.0(2) OS 113.9(2) C7 Cll C13 129.8(2) OS 109.7(2) Cll C13 120.9(3) 106.2(2) C12 02 122.2(2) Cl CS C4 96.6(2) C12 Cll 109.7(2) Cl CS CS 115.8(1) C12 Cll 128.0(3) Cl CS CIS 115.3(2) C15 C5 110.2(1) C4 CS CS 112.0(2) CIS 04 122.8(2) C4 CS CIS 106.4(1) C16 C17 112.1(2) C6 CS 109.8(2) 04 CIS C17 125.1(2) CS C6 C7 116.0(2) 05 C18 06 123.0(2) C6 C7 C8 114.0(1) 05 C18 C19 111.4(2) C6 C7 Cll 111.3(2) 06 C18 C19 125.6(3) C8 C7 Cll 102.5(2)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 6. Bond Angles in Degrees Compound 3 (cont.)

Angle Angle tllLl = ~ L = ===L= C8 02 C12 111.0(2) C7 Cll 114.8(2) C2 Cl CS 103.9(1) C8 C7 Cll 101.3(2) C2 Cl CIO 116.8(2) 02 C8 C7 10S.9(1) C5 Cl CIO 120.9(2) 02 C8 C9 105.3(2) Cl C2 C3 104.7(1) C7 C8 C9 118.5(2) C2 C3 C4 10S.2(1) C8 C9 CIO 115.5(2) 01 C4 C3 111.7(2) Cl CIO C9 111.0(2) 01 C4 CS 114.0(2) Cl CIO C14 115.7(2) C3 C4 CS lOS.O(l) C9 CIO C14 109.9(2) Cl C5 C4 98.0(1) C7 Cll C12 104.7(2) Cl C5 C6 115.2(1) C7 Cll C13 118.8(2) C5 CIS 113.1(1) C12 Cll 112.8(2) C5 C6 111.3(1) 02 C12 121.4(2) C4 CS CIS 110.1(1) 02 C12 110.0(2) C6 108.8(1) 128.6(2) C5 C7 114.4(2) 112.5(2) C6 C8 112.9(2)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 7. Selected Torsion Angles ( ) Comp

Atoml Atom2 Atom3 Atom4 Angle

C12 02 C8 C7 9.8(4)

C8 02 C12 Cll -2.0(3)

C5 Cl C2 C3 -30.1(3)

C2 Cl 44.8(3)

-63.8(3)

Cl 46.2(4)

Cl -81.9(4)

C2 C4 2.4(4)

C2 C3 01 150.7(3)

C2 C3 C5 26.4(3)

C3 C4 Cl -43.3(3)

Cl C6 C7 -20.0(4)

Cl C5 CIS 04 -86.3(3)

C5 C6 C7 C8 68.6(3)

C6 C7 C8 C9 -10.9(4)

Cll C7 C8 02 -12.9(4)

C8 C7 Cll C12 12.1(4)

C8 C9 CIO Cl 44.6(4)

C7 Cll C12 02 -6.9(5)

0C13 Cll C12 03 10.1(8)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 7. Selected Torsion Angles ( ) Compound 2

Atoml Atom2 Atom3 Atom4 Angle

C12 01 C8 C7 11.1(2)

C8 01 C12 Cll -4.8(3)

C5 Cl C2 C3 -33.5(2)

C2 Cl C5 47.1(1)

CIO Cl C5 -60.2(2)

C5 Cl CIO 42.1(2)

C5 Cl CIO C14 -85.8(3)

Cl C2 C3 C4 5.3(2)

C2 C3 C4 05 146.2(2)

C2 C3 C4 C5 25.3(2)

C3 C4 C5 Cl -44.0(1)

Cl C5 C6 C7 -22.0(2)

Cl C5 03 -79.7(2)

C5 C6 C7 C8 68.7(3)

C6 C7 C8 C9 -10.4(4)

Cll C7 C8 01 -12.4(2)

C8 C7 Cll C12 10.2(2)

C8 C9 CIO Cl 46.8(2)

C7 Cll 01 -4.0(3)

C13 Cll 02 0.7(4)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 7. Selected Torsion Angles (°) Comp

Atom2 Atom3 Atom4 Angle

C12 02 C8 C7 14.2(2)

C8 02 C12 Cll 2.8(2)

C5 Cl C2 C3 -32.8(1)

C2 Cl C4 46.7(2)

CIO Cl C6 -61.6(2)

C5 Cl C9 41.0(2)

C5 Cl CIO C14 -85.0(2)

Cl C2 C3 C4 5.0(2)

C2 C3 C4 01 148.6(2)

C2 C3 C4 24.6(2)

C3 C4 C5 -43.5(2)

Cl C5 C6 C7 -21.6(2)

Cl C5 CIS 04 179.5(1)

C5 C6 C8 73.8(2)

C6 C7 C8 -18.6(2)

Cll C7 C8 -24.2(2)

C8 Cll 25.4(2)

C7 C8 C9 CIO -66.4(2)

C8 C9 Cl 48.3(2)

C9 CIO C14 177.5(2)

C7 Cll C12 02 -18.7(2)

C13 Cll C12 03 32.6(3)

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 5.1. The molecular structure of 15 acetylrudmollin (5.1), with thermal ellipsoids drawn at the 30% probability level, and H atoms represented by circles of arbitrary

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 5.2. The molecular structure of diacetylrudmollin (5.2), with 30% ellipsoids.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Fig. 5.3. The molecular structure of llocH, 13-dihydrorudmollin (5.3), with 50% ellipsoids.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. THE SYSTEMATICS OF THE GENUS RUDBEOCIA AND BIOLOGICAL ACTIVITIES OF RUDBECKIA CONSTITUENTS

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ABSTRACT-Sesquiterpene lactones, polyacetylenes and lignanes are present in Rudbeckia species. There are many varieties and the range of biological activity is broad. Various sesquiterpene lactones and lignanes found in Rudbeckia are known to exhibit antitumor activity and to inhibit certain enzymes. This chapter summarizes the present state of knowledge on the chemistry of the genus Rudbeckia.______

INTRODUCTION

Members of the genus Rudbeckia are commonly referred to as cone

flowers. Rudbeckia belong to the tribe Heliantheae (Asteraceae), and

members are found as perennial or annual herbs. This new-world genus was

named in honor of Rudbeck (1630-1702) and his son Olaf (1660-1740) from

Upsala, Sweden. They were predecessors of Linnaeus. The genus was

described by Rudbeck. It now is recognized to contain two sections or

subgenera Rudbeckia and Macrocline. The two subgenera have different

chromosome numbers. The basic chromosome number of the subgenus

Rudbeckia is n=19II whereas Macrocline has n=18II chromosomes. In the

southeast United States six species of Rudbeckia grow. R. triloba is

rare in Louisiana.

Sesquiterpene lactones are very frequently found in the Asteraceae.

This group of terpenoids has attracted much interest due to their varied

biological activities [1]. Their general toxicity has contributed to the

ecological success of the Asteraceae family in its competition with

other plants and in expanding its range. These compounds are also useful

systematic characters within the Asteraceae. Similar roles are

attributed to other natural products, such as lignanes, in other plant

families.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Sesquiterpene lactones, polyacetylenes, and lignanes can be used for

the taxonomic characterization. Certain features make these compounds

useful and allow them to fulfill the major requirements of suitable

analytical and systematic markers. They show structural diversity and

they undergo diverse chemical transformations faciliting structural

analysis. Among the many natural products in the Asteraceae,

sesquiterpene lactones are especially important in biochemical

systematic studies. These commonly found lactones have been used as

markers in studies of taxonomic variations. They serve as a vehicle for

the exploration of the links between distribution trends of similar

compounds and proposed biosynthetic relationships.

Biological activity studies of different types of sesquiterpene

lactone also contribute to the understanding of the ecological functions

of these compounds in plants. However, the detection or failure to

detect sesquiterpene lactones in a particular species must be considered

in the context of the family’s total terpene chemistry.

RESULTS AND DISCUSSION

A. Taxonomic implications of Secondarv Metabolites in the Genus

Rudbeckia

The two subgenera of Rudbeckia differ in the chromosome number and

in the type of secondary metabolites found. Since the sesquiterpene

lactones in Rudbeckia vary from species to species, it should be

possible to identify a species by means of this variation. Table 1 lists

the species and subspecies in Rudbeckia. Thus far, only ten species of

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Rudbeckia have been investigated chemically. These investigations are

summarized in Table 2.

B. Chemical distribution

Chemical investigation of the genus Rudbeckia resulted in the

discovery of three major chemical groups: sesquiterpene lactones,

polyacetylenes and lignanes.

Early work on the chemistry of Rudbeckia documented the following

distribution of compounds:

1. Sesquiterpene lactones of the pseudoguaiano1ide type have been

reported from R. mollis Ell, [2] and from R. laciniata [3,4];

2. Flavonoids were isolated from R. fulgida Ait [5], R. hirta [6, 7,

8] and R. mollis [9];

3. Carotenoids have been reported from R. fulgida [10];

4. Polyacetylenes were detected in the roots of R. triloba [11],

R. hirta [12,13] and in aerial parts of R. fulgida [14], R.

laciniata [13], and R. nitida [13].

Fig 1-7 shows the structures of the compounds previously isolated from

Rudbeckia.

Table 2 lists those species and varieties that have been chemically

examined along with the structural types of the compounds found in each

taxon. This table illustrates the usefulness of sesquiterpene lactone

investigations for plant classification. The presence or absence of a

particular structural type is useful as a character and should be

considered along with morphological and cytological characters of

significance. In this example flavonoids and sesquiterpene lactones

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. occur predominantly in R.mollis and R. subtomentosa from the subgenus

Rudbeckia whereas lignanes occur predominantly in R. scabrifolia, R.

maxima, and R. nitida from the subgenus Macrocline.

The examination of the chemistry of seven Rudbeckia species resulted

in the identification of 42 compounds. Thirtyfour of these compounds are

natural and eight are synthetic derivatives. Of the 34 natural

compounds, ten were previously unreported. The compounds isolated were

16 pseudoguaianolides, three germacrolide, six eudesmanolides, one

flavonoid, two lignanes, two triterpenes, five sesquiterpene esters, two

polyacetylenes and one triterpene. In addition, trans-trans-farnesol,

caryophyllene A, caryophyllene A oxide and squalene [15,16,17] were

This chapter reports natural products from Rudbeckia which belong to

three basic skeletal types of lactones (germacrolides, eudesmanolides,

pseudoguaianolides) as well as polyacetylenes, triterpenes, lignanes and

sesquiterpene esters. The structures and their stereochemistry were

elucidated mainly on the basis of ^H, NMR spectroscopy as described

in previous chapters. In six cases X-ray diffraction studies aided the

elucidation or confirmed the proposed structure.

C. Species relationships within Rudbeckia

Polyacetylenes are the most common group of compounds present in

Rudbeckia species. R. fulgida Aiton is unique in that carotenoids,

labdane and benzoquinone have been reported only from this species.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 164

Sesquiterpene esters have been reported only from R. grandiflora

Sweet [15]. Two species, R. subtomentosa Pursh and R. mollis, contain

triterpenes. On the other hand, flavonoids, sesquiterpene lactones, and

lignanes have been isolated from four different species of the subgenus

Rudbeckia.

As might have been predicted, R. grandiflora, which was placed in

the subgenus Rudbeckia on the basis of morphology, is chemically more

similar to R. subtomentosa and R. mollis from the subgenus Rudbeckia

than to R. nitida Nutt, and R. triloba L. from the subgenus Macrocline.

R. grandiflora, R. subtomentosa and R. mollis typically contain

pseudoguaianolides. Leaves of R. grandiflora and R. subtomentosa contain

the same pseudoguaianolides ligulatine C and desacetyl1igulatine C which

suggests a close genetic relationship between these two taxa.

Three species of the subgenus Macrocline: R. maxima Nutt, R.

scabrifolia Brown, and R. texana are distinguished by the predominant

presence of lignanes and the absence of sesquiterpene lactones. R.

triloba of the subgenus Rudbeckia also contains ligananes and no

sesquiterpene lactones suggesting it might be better clasified in

Macrocline. However, examination of the crude syrup of R. maxima

indicated the possible presence of sesquiterpene lactones which could

not be isolated and identified because of the limited amount of material

available.

The results reported in an earlier chapter suggest that the roots of

Rudbeckia grandiflora can be characterized chemically by germacrolides.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 165

its leaves by pseudoguaianolides and its flowers by sesquiterpene

D. Agricultural, medicinal, pest repellent, and allelopathic

implications of biologically active secondarv metabolites present in

Rudbeckia

Lignanes;

It is known that lignanes have a variety of pharmacological actions

in man, which include hypertensive and sedative effects [18]. There is

evidence that lignanes play a role in plant-fungus, plant-plant and

plant-insect interactions [19]. The lignanes, pinoresinol dimethyl

ether, and yangambin, isolated from Rudbeckia, are known to show

antiviral activity, and inhibit cAMP phosphodiestarase [19]. Pinoresinol

dimethyl ether is an inhibitor of microsomal mixed function oxidase

activity [20].

Sesquiterpene lactones:

Sesquiterpene lactones from Rudbeckia deter the grazing of sheep and

cattle and in few cases have been responsible for severe livestock

losses [21]. In some instances the ingestion of R. laciniata is believed

to have been the cause of death in hogs, swine, sheep and cattle

[22,23],

Rudbeckia has been listed as one of the Heliantheae genera causing

contact dermatitis [24]. Rudbeckia includes several different species

that have been or are being used for medicinal purposes: R. hirta L. has

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. been used to treat skin infections. A study has shown that extracts from

the plant have antibiotic properties [22,23]. The sesquiterpene

lactones, rudmollin and 15-acetylrudmollin, show antitumor activity [2]

isoalantolactone and 4,5-endoisoalantolactone, isolated from R. mollis

Elliot, show ant-repeilent activity [25,26].

Polyacetylenes:

Polyacetylenes have been used in Africa and Asia to treat skin

diseases [27]. Some polyacetylenes are known to be very active against

fungi, bacteria, and nematodes [28]. Polyacetylenes are also phytotoxic.

They are found in fresh water and marine algae [29]. The polyacetylenes

thiarubrine A and thiophene B, isolated from R. mollis, exhibit

phototoxic [30] and antifungal activity [31].

Flavonols:

The flavonol eupatolin is a rare flavonol glycoside which was first

isolated from Eupatorium 1ignstrinnm DC. [32] and later from Hymenoxys

subacaulis [33], R. mollis [9] and R. subtomantosa [16]. In R. hirta,

the yellow flavonols serve as insect nectar guides [8].

CONCLUSION AND PROSPECTS FOR FUTURE RESEARCH

Our comparative chemical studies of the subgenera Rudbeckia and

Macrocline suggest a distinct chemical division between Macrocline and

Rudbeckia. Macrocline contains lignanes as the major constituent whereas

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 167

sesquiterpene lactones are the major components in the subgenus

Rudbeckia.

The chemical evidence presented in this chapter corroborates the

distinct boundary between Macrocline and Rudbeckia which was based on

morphological data. For example, R. grandiflora is characterized by a

chromosome number 19 and by the pseudoguaianolides rudbeckin A,

ligulatine C, and desacetyl1igulat ine C. In contrast R. maxima has a

chromosome number 18 and produces the lignanes pinoresinol dimethyl

ether, and yangambin, but sesquiterpene lactones were not detected.

The two subgenera of the genus Rudbeckia have not been investigated

so thoroughly to warrant the assertion that the presence of

sesquiterpene lactones and the presence of lignanes are mutually

exclusive. However, it does appear that most members of the subgenus

Macrocline do not synthesize sesquiterpene lactones. The capacity to

synthesize germacrolides, eudesmanolides, and pseudoguaianolides is a

characteristic of the subgenus Rudbeckia

A more intensive chemical sampling is needed to resolve taxonomic

problems associated with the Rudbeckia complex. Search for new secondary

metabolites from unexplored Rudbeckia species is needed. They must be

investigated by means of a variety of approaches.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1. Taxonomy of Rudbeckia

Family: Asteraceae Tribe:Heliantheae Genus: Rudbeckia

Section:Rudbeckia species: R. fulgida Aiton var. fulgida palustris rupestris spathulata sullivantii umbrosa R. graminifolia T. & G. R. grandiflora Sweet var. alismaefalia war.grandiflora R. heliopsidis T. & G. R. hirta L. var. ar^ustifolia

pulcherimma R. missauriensis Engelm R. mollis Elliot R. triloba L. var. pinnatiloba rupestris

R. subtomentosa Pursh

Section: Macrocline species R. auriculata Perdue R. califarnica A.Gray var califarnica intermedia

R. laciniata L. var. ampla bipinnata digitata humilis laciniata R. maxima Nutt R. mohrii A. Gray R. nitida Nutt var.

R. occidental is Nutt ' alpicola mantana occidental is R. scabrifolia Brown

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 2 Distribution of secondary metabolites in Rudbeckia Species.

P F+ s§ l " others Ref

labdane & 5.10 R.fulgida •• benzoquinone 14

R. hirta •• 6 R. triloba • * 12, 13

R. nitida • * 16

R. scabrifolia *

R. maxima *

R. laciniata •• 3,4

R. grandiflora esters 15 var.alismaefolia

R. subtomentosa * a triterpenes 16

R. mollis * • • triterpenes 2,17

Polyacetylenes ^ Flavonoids * Carotenoids ^ Sesquiterpene lactones " Lignanes

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1. Adams, R. and Herz, W. J. Chem. Soc. (1949) 71, 2546.

2. Herz, W. Kumar, N. (1981) J. Org. Chem. 46, 1356.)

3. Bohlmann, F. Jakupovic, J., Zdero, C. (1978) Phytochemistry 17,

4. Jakupoyic, J. , Jia, Y., King, M. R., Bohlmann, F. (1986) Liebis Ann. Chem. 1474.

5. Herz, W., Kulanthaiyel, P. (1985) Phytochemistry 24, 89.

6. Waddell, T. G., Elkins, S. K., Mabry, M. A., Singri, B. P., Wagner, H., Seligman, Herz, W. (1976) Indian J. Chem. 14B, 901.

7. Jauhari, P. K., Sharma, S. C., Tandon, J. S. and Dhar, M. M. (1979) Cent. Drug Res. Inst. 18, 359.

8. Thompson, W. R., Meinwald, J., Aneshansley, D. and Eisner, T. (1972) Science 177, 528.

9. Iyengar, M. A., Singri, B. P., Wagner, H. , Seligman, 0.and Herz, W. (1976) Indian J. Chem. 148, 906.

10. Valadon, L. R. G., Mummery, R. S. (1971) Phytochemistry 10, 2349.

11. Bohlmann, F. Grenz, M., Wotschokowsky, M. , Berger, D. (1967) Chem. Ber. 100, 2518.

12. Bohlmann, F. and Kleine, K. (1965) Chem. Ber. 84, 3081.

13. Atkinson, R. and Curtis, R. P. (1965) Tetrahedron Lett. 5, 297.

14. Herz, W. Kulanthaiyel, P. (1985) Phytochemistry 24, 89.

15. Vasquez, M., Macias, F. A., Fischer, N. H. (1988) Phytochemistry 27. 2198.

16. Vasquez, M., Quijano, L., Macias, F. A., Fonczek, F., Urbatsch, L., Cox, P. B., and Fischer, N. H. (1989) Phytochemistry submitted.

17. Vasquez, M., Quijano, L. and Fischer, N. H. (1989) Phytochemistry submitted.

18. Nikaido, T., Ohmoto, T., Kimoshita, T. , Sakaura, U., Nishibe, S. and Hisada, S. (1981) Chem. Pharm. Bull. 29, 3586.

19. Mac Rae, W. and Towers, N. G. (1984) Phytochemistry 23, 1207.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 20. Weidenhamer, J. Personal communication.

21. Pieman, A. K. (1986) Biochem. Svst. Ecol. 14, 255.

22. Steyermark, J. A. (1963) Flora of Missouri. The Iowa State University Press Ames, Iowa, U. S. A. p. 99.

23. Cox, D. D. Common Flowering Plants of the Northeast. State University of New York Press, Albany, p. 171.

24. Rodriguez, E., Yoshika, H. and Mabry, I. J. (1971) Phvtochemistrv 10, 1145.

25. Okunade, A. L. and Wiemer, D. F. (1985) Phvtochemistrv 24, 1199.

26. Hubert, T. D., Okunade, A. L. and Wiemer, D. F. (1987) Phvtochemistrv 6, 1751.

27. Wat, C-K. , Johns, I. and Towers, G. H. N. (1980) Ethnopharmacolcgy 2, 279.

28. Bohlmann, F., Burkhardt, T. and Zdero, C. (1973) Naturally Occurring Acetylenes, p. 547, Academic Press.

29. Arnason, T., Stein, J. R., Graham, E. , Wat, C-K. and Towers, G. H. N. (1981) Can. J. got. 59, 54.

30. Towers, G. H. N. (1987) Plant Phvsiol. 6, 85

31. Rodriguez, E. Aregullin, M., Uehara, M. , Wrangham, R., Nishida, T., Abramowski, A., Kinlayson, A. and Towers, G. H. N. (1985) Experientia 41, 419.

32. Quijano, L., Malanco, F., Rios, T., (1970) Tetrahedron 26, 2851.

33. Iyengar, M. A., Katti, S. B., Wagner, H. , Seligman, 0. and Herz, W. (1975) Arogya J. Health Sci.. 1, 74.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. .OH

HO" -OH

HO" kOH

HO'

REFERENCE:

Valadon.L.R-G.,Mummery,R.S.Phytochemistry (1971) 10 2349.

Fig. 6.1. Carotenoids isolated from fiudbeckia species.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. R 4

1’ R 3ON

R ^ '"OR 5 O H 0

R, R 2 R 3 R 4 R s S o u r c e R e f . H O M e M e OHH R . h i r t a 6 H O M e M e O M e H R . h i r t a 6 H O M e M e HH R . h i r t a 6 O M e H M e H R . h i r t a 7 H O M e M e OH G l u R . h i r t a 8 H O M e G lu H H R . h i r t a 8 H O H G lu H H R . h i r t a 8 HH HOH R t i o m R . f u l g i d a 5 H H H O H G l u R . f u l g i d a 5 H H R h o m H H R . f u l g i d a 5 HH H H G l u R . f u l g i d a 5 H H G l u H H R f u l g i d o 5 H O M e M e OH R . m o l l i s 9

Fig. 6.2. Flavonoids isolated from Rudbeckia species.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Me'C=C-Cy|-C=C-]-CH=CH2 triloba S I iz /?. fuigida

HOCHgC^C— ^ Ç ^ C = C^CH = CH 2 triioba

R. hirta Me-C = C— ^f3^C=C-PcH=CH^H Ft- laciniata ^ ^ R. nitida

Me-C = C — ^ ~ ^ C = C-f CH = C h ] ^ H R. hirta

M e -[ C = C ]^ ^ ^ C = CCH = CH 2 fulgida

Me-[c = C-]-

Fig. 6.3. Polyacetylenes isolated from Rudbeckia species.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 2 a : R,= R ,= H

2 b : R,= Ac Rg= H

2 c : R,= H R2= A c

Herz, W. and Kum ar,N.(I98I),J. Org. Chem; 4 6 . 1356

Fig. 6.4. Sesquiterpene lactones isolated from Rudbeckia mollis.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Jakupovic, J., Jia, Y.,King,M. R.ond Bohlmann, F.,(1986), Liebigs Ann. Chem., 1474

Fig. 6.5. Secondary metabolites isolated from roots of Rudbeckia laciniata.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. H j C [ c s c ] g C H : C H g

1 - R = H 2 : R = OH

Bohlmann,F., Jakupovic ,J. and Zdero, C., (1978),Phytochemistry 17, 2 0 3 4

Fig. 6.6. Secondary metabolites isolated from aerial parts of Rudbeckia laciniata.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. M eO, CHO .OMe

CO,H

H OH

.OH HO H •CO.H H OH

5a:R=0Ac = H "R 5b ; R = R, = OAc 5c : R = OH R ,= H

Herz W. and Kulanthoivel.P.(1985),Phytochemistry , 2 4 ,1 .8 9 .

Fig. 6.7. Secondary metabolites isolated from Rudbeckia fulgida.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. s i II

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. il

JPo il

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. R . f u l g i d a

R. grandifioro

Cox, R B., U rbofsch, L.E. (1988) Louisiana Academy of Sciences 61, 0 0 0 0 .

Fig. 6.10. Geographical distribution of Rudbeckia fulgida and Rudbeckia grandiflora In the southeastern United States. 9 S

R . m o l l i s

R. subtomentosa

C ox, R B., U rbotsch, L.E. (1988) Louisiana Academy of Sciences 61, 0 0 0 0 . Fig. 6.11. Geographical distribution of Rudbeckia mollis and Rudbeckia subtomentosa in the southeastern United STRUCTURE ELUCIDATION OF SECONDARY METABOLITES FROM RUDBECKIA SPECIES BY SPECTROSCOPIC TECHNIQUES AND REVIEW OF SESQUITERPENE LACTONES

A Dissertation

Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of

Doctor of Philosophy in The Department of Chemistry

by Marta Vasquez Universidad de Antioquia, Colombia, 1971 ., Universidad del Valle, Colombia, 1981 August 1989

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. INTRODUCTION

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 184 This review summarizes the progress made in the field of

sesquiterpene lactones in 1986 and 1987. Included are data from

reports of isolation and structure elucidation.

In the relatively short time since the last review article on

sesquiterpene lactones (as defined by Holub, M. et al. [109]) was

published in 1985 [74] about 522 sesquiterpene lactones have been

reported from 173 sources. The new lactones include two new skeletal

types, the vernojalcaguaianolides and the jalcaguaian-olides and two

angelate-derived side chains. By the end of 1985, a total of 2650

different sesquiterpene lactones had been described. To date there

have been close to 3200 structures of sesquiterpene lactones

elucidated and, due to modern instrumentation and techniques, the

number is rapidly increasing. The highest rate of reporting new

compounds occurred during 1984 and 1985 (Figure 1-1). During 1986 and

1987 the rate has been decreasing, probably because attention has

been shifting to the discovery of a broad spectrum of biologically

active compounds as well as to the study of their pharmaceutical use

and ecological functions. Several sesquiterpene lactones have

attracted considerable attention due to their various biological

activities such as antitumor activity, cytotoxicity, allergic contact

dermatitis, and microbial and plant growth inhibition [194].

Sesquiterpene lactones form one of the largest group of

cytotoxic and anti-tumor compounds of plant origin. Most of these

active sesquiterpene lactones are found in species of the Asteraceae

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1 8 5

although some of them originate in the Magnoliaceae, Apiaceae and

even fungi [69]. Most active sesquiterpene lactones (such as

a-santonin (I) [208]) show cytotoxic activity against the growth of

KB cells in vitro and activity against in vivo P38B leukemia [194].

Sesquiterpene lactones of this type are represented by compounds II

and III [19]. However, because compounds closely related in structure

often differed in their activity, no clear structure versus activity

relationship has been found [69]. These studies suggest that in

addition to the a-methylene-y-lactone structure, other factors are

important for cytotoxicity. Despite the expanding data base, and

because of the rapid expansion of the information available, the task

of abstracting data and reporting it in a manageable form is vital to

the as yet unsolved problem of developing a comprehensive system for

applying sesquiterpene lactones to the structure-activity

relationship.

The wide variety of chemical structures so far discovered is

mirrored by a diversity of biological activities. Compounds that have

antifeedant (such as alantolactolactone (IV), helenalin (V), and

bakkenolide A (VI) [171]), bio-antimutagenic (such as dehydrocostus-

lactone (VII) [145]), anti-bacterial such as confertin (VIII) [64]

anti- secretory (such as parthenolide (IX) and canin (X) [93]) and

anti-fungal (such as parthenolide (IX) [83]) properties are widely

distributed. Various sesquiterpene lactones are known to be toxic to

human and animal parasites, insects and vertebrates. Many of the

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1 8 6

compounds or plants containing them cause allergic contact dermatitis

in humans .

Sesquiterpene lactones also act as plant growth regulators and

are responsible for allelopathic properties of many plants. Various

activities of sesquiterpene lactones suggest their evolutionary

significance in plants as deterrents protecting against attacks by

herbivores, fungus and bacteria. Allelopathic agents such as

chlororepdiolide (XI) have been reported [240].

The following is a brief description of reviews published for

the last seven years. Two reviews dealing with sesquiterpene lactones

of pharmacological interest were published in 1981. One of these [28]

reports 35 germacranolides such as costunolide (XII) and epitulipin-

olide (XIII) and the other reports 20 eudesmanolides such as

alantolactone (IV) and arbusculin B (XIV) [28]. In 1981, Romo de

Vivar [204] reviewed the mexican research into sesquiterpene

lactones. The review contains a brief discussion of the structure of

92 substances discovered by mexican scientists.

Cladistic or Hennigian phylogenetic systematics is reviewed by

Seaman et al. [227] in 1983. It suggests that a reaction sequence by

which a given structure might have been derived from a postulated

precursor can be derived. The probability that the hypothesis

represents the reality can be increased if a series of compounds, all

occurring together in a single organism, can be shown to be

constituent parts of the hypothetical pathway. In 1985 a review on

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1 8 7

the evolution of sesquiterpene lactone in angiosperms [69] was

published. It outlined the evolutionary trends accompanying

sesquiterpene lactones in the angiosperms.

In the same year, a review was published covering in vitro and in

vivo bioassay tests of sesquiterpene lactone allergenic activity

[256]. Also, Hoffmann et al. [106] reported progress in the synthesis

of the a-methylene- y-lactone ring after a description of the

biological activities.

In 1986, Herz [101] reviewed biogenetic aspects of sesquiterpene

lactone chemistry and Holub et al. [Ill] published a review which

contains 107 sesquiterpene lactones in the Umbelliferae. The review

suggests that the biogenetic steps leading to the sesquiterpene

lactone in the Umbelliferae and the Asteraceae (Compositae] form two

parallel series and proceed by similar reactions.

In 1986, Pieman [194] published a review which summarizes the

biological activities of sesquiterpene lactones and their structure-

activity relationships.

The primary focus of this review is directed toward recent

advances in knowledge concerning naturally occurring sesquiterpene

lactones with a secondary focus upon some related important studies

of new synthesis. The number of naturally occurring compounds

reported during the past two years by skeletal types of sesquiterpene

lactones are recorded in table 1-2. The natural products are listed

under the name used in the original literature. All taxa from which

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1 8 8

sesquiterpene lactones have been reported are listed together with

the compound names, and the literature cited. Structures for all

reported compounds are included. In order for this review to serve as

a useful reference, two appendices are included: one listing

alphabetically the natural sources from which sesquiterpene lactones

have been isolated (Appendix A), the second listing the names and

structure number of all reported compounds (Appendix B). This latter

appendix includes for each compound the page number of the first

report in table 1-1. However, as any human activity this one is not

free from errors and omissions, corrections and suggestions for the

improvement of this review will be appreciated. It is hoped that this

review will stimulate new experimental work, its use will suggest

changes in the organization and presentation of the information that

will eventually allow the researcher rapid access to information that

will illuminate some hitherto unseen relationship and thus encourage

the advancement of science.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1 8 9 Table 1.2. Number of sesquiterpene lactones by skeletal type

trans.trans Germacranolides 72

Melampolides 39

Heliangolides 39

cis.cis Germacranolides 83

Furanoheliangolides 3

Seco-germacranolides 3

Elemanolides 13

Eudesmanolides 71

Eremophylenolides 11

Guaianolides 120

Seco-guaiano1 ides (xanthanolides) 2

Pseudogua i ano1 ides 29

Seco-pseudoguaianolides 11

Vernomargolides 4

Vernojalcanolides 5

Scorpiolides 1

Picrotoxane 1

Daucanolides 1

others 14

Total 522

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Co i A 0 II

W = » III IV

H i t \L

0 (5h ) r °

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of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission s 0 fü § s s

i i =

I i

It l à •§LÏ r*P M o'— (% 1% 1% O

s 8 d & XXX

5 S 3- X X 5 5 1 8 1s 1 g 8 II X X X X

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s §

J 11 t, H M H s S 5

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ilT -I I

J s I I I I*

5 X X

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8^-lsobutyryloxy-9/î-hydroxy-10- Blcinvilleo lotifolic acetoxymethylgermacrodien-6.7-olide

8/?-Me t hy I bu ty r y I oxy-9^-hydr oxy-10- BloinviI lea (allfolia ocetoxyme thy I gerrxicr ad i sn-6,7-01 i de

Ang Jurinelloide Jurinella moschus

Ang-5-CH 20-Hydroxyjurinelloide JurineI la moschus

*8-Oxo-2a-9-dihydroxy-tron3,trons- Tonocctum vulgare germacro-l(IO),4-dien-trans-6,12-

*No stereochemistry Is given at C-9 5 5

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. structure Substituent Name of Compound Plant Source Ref

1ip,13-EpoxyBpHulIplnolld8

OAc 3f-Ac»*oxy-1If,13-*poxy- epliulIpinolIds

* Isodeoxyelephantopin Elephonlopus acaber 265

«Compound was previously reported without stereochemistry Substituent Name of Compound

• 13-OlhydrodBOxynlephanfopIn Elaphanfopus scobar 265

"Compound was previously reporfed wllhoul

slereochemlslry

2-ept-0aoxyorlhopapp-4E-anolIda Elaphanlopus tnol lia 121

Totnenphcniopin B Elaphanlopus lomentosus 99

Tomenphanfopln / Elaphanlopus fcmenfosus 99 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. I I

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. structure Substituent Name of Compound

3-Acetylchamissonin

Gochnotia hypoleuco

Tig Gochnatla hypoleuco I I i-But Gochnotia hypoleuco

U-Acetoxydesocetyl lourenobiolide Perymenium mendezii Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 1! II II il Il 11 liil

§ i g

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Name of Compound Plant Source

DoacolyIfulIpînolIda-ip,lOa-apoxJde Colulo cinereo 159

la, 10/J-Epoxyovo»Hol In GroenmanlaMo roslnosa 264

8a,15-fl!s(acefoxy)onhydrowarloforln Vernonia diffusa

15-AcefoxydenfafIn B Vernonia diffusa

0. \cOH

01-Vo I H 8/Î-1 sova I ary I oxy-10o-hydroxy-1-oxo- germacra-4E,11(13)-dIana-12,Ga-o 11 da

Bfi-1 sova I ary I oxy-3/î, 10a-d Ihydroxy- 1-oxo-garmacro-4E,11(13)-dlana- 12,6a-o1lda 208

I s E

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. structure Substituent Name of Compound Plant Source

Calalna E Calao zacotachlchl

Calao zaeofaehlchl

Vernopoppol Ida Vernonia chloropoppa 140

Varnopappollda-8-O-malhacrylola Varnonlo chloropoppa 140

Varnopoppollda-8-O-llglofa Varnonlo chloropoppo 140

6/J-HydroxyvornopoppoHda-8-0- Varnonlo chloropoppo 140 malhacrylota

,..>‘OAng

VarnopotensolIda 8-O-ongalale Varnonlo palans I I

lu I {{

5 5 5^ 5 1 ^

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t !

5 5 5 5

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I 5

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. II

il "

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Substituent Name of Compound Plant Source

Eupc'tor ium quadrangular

Austroliobum candidum 125

H H Tig 13-Oeacetylmarginatin Vernonia marginata

Ac H Ac 8-Oesacylmarginotin 8-O-acetote Vernonia jalcono

Ac OAc Ac 2a-Acetoxy-8-de3acylglaucollde-E- Muschlerio atoizi i

Ac OAc Mac-4-OH 19-Hydroxyglaucolide E Vernonia cinereo 2

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Substituent Name of Compound Plant Source

Moc Orthopoppolide methacrylate Elephontopus ongustifolius

Sen Orthopappolide seneclaate Elephontopus ongustifolius

Tig Orthopappolide tiglote Elephontopus ongustifolius

DesocyIisoelephantopi n seneci cate Elephontopus ongusti folius

DesocyIisoelephontopin tiglote Elephontopus ongusti folius

Polymnio moculoto 153 Plant Source structure Substituent Name of Compound

OMebut

DesocyHagitinin C-8-0-f2- methylbutyrotej

If.10fl;4a,5f-Diepoxy-Bf-isobutoxy Smyrnlum perfoliolum 90 glechorronol ide

If,10a;4a,5f-Dlepoxy-8a-i3obutoxy Smyrnium perfoliotum 90 glechomonolide

•No stereocliemistry given ot C—4 I I

I.L r II fl I

s i

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ill

1 I I

k à à

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. i 2

Il |]

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Substituent Name of Compound Plant Source

Mac OAc Ac 14-O-Acetylvernocislifolide-8-O- Vernonio cisti methocrylote

14-O-Acetylvernocistifol ide-Q-O- Vernonio cisf

14-O-Acetyivernocistifolide-8-O- Vernonia cisti (2,3-epoxyisobutyrote)

14-O-Acety I vernoci st i fol ide-8-O- Vernonia cisti ongelote

Mac OAc Sen 14-0-Senecioylvernocistifo1ide-8-O- Vernonia cisti methacrylate

Ang OAc Sen 14-O-Seneciaylvernocistifolide-&-0- Vernonia cisti ange late

Epo OAc Sen 14-O-Senecioylvernocistifoiide-8-O- Vernonia cisti (2,3-epoxyisobutyrate)

Ang OAc Sen-CH 14-0-(4-HydroxysenecioyI)verno- Vernonia cisti ci st i folide-B-O-angelote

Epo OAc Sen-CH 14-0-(4-Hydroxysenecioyl)vernocistif- Vernonia cisti ol ide-8-0-(2,3-epoxyisobutyrate)

Mac OAc Sen-CH 14-0-(4-HydroxysenecioyI)vernocistif- Vernonia cisti olide-8-O-methocrylate

Mac H Ac 14-0-Acetyl-9-desacetaxyvernocistif- Vernonia cisti olide-B-O-methocrylote

Ang H Sen-CH 9-0esacetoxy-14-O-(4-hydraxysenecioyl) Vernonia cisti vernocist ifolide-B-O-ongelote

Ang H Sen 9-Desocetoxy-14-O-senecioylverno- Vernonia cisti ci st i fol ide-8-O-ongelate Substituent Name of Compound Plant Source

,OSen-OH JDAc

14-0-(4-Hydroxy3enecioyl)isoverno- ci st i fol ide-8-O-angelote 'OAc

'OH

Mac lO-Oesocetoxy-9f-fiydroxyg 1 auco 1 ide A Vernonia chloropoppa 140 )Ac Ang - StiIpnotomentolide 8-0-angelote Vernonia patens

Tig OAc - S-Oesocylgloucolide A-tiglate Vernonia erdverbengii

'OAc 10,14-Dehydro3t!Ipnotomentolide- Vernonia marginata 8-O-t iglote Name of Compound Plant Source

OMac-4-OH Vernoc ineroI ide-S-O-f't-hydroxy- Vernonia cinereo methocry lotej

8a-(2-MethyIocry IoyIoxy )- Vernonio patens 135 compact if lorIde

8a-AngeIoyIoxycompoct if loride Vernonio patens 135

OMac-4-OH

Et (Z) BrochycolyxolIde Vernonia brochycolyx 140

16,17-Oihydrobrachycalyxolide Vernonia brochycolyx 140 I!

w I I

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Substituent Name of Compound Plant Source

O i-B u t

H Me Desoxytagi t inin B-3-O-nœthy I ether Greenmonlella resinoso 264

OMe H 2^-Wethoxydesoxylagitinin B [sic] Greenmaniella resinosa 264

H Mebut Desacyltagilinin F-8-0-[2- Greenmaniello resinosa 264 methylbutyrote]

Me i-But Tog!tinin F-3-O-methyI ether Greenmaniella r

2 ’,3'-0ihydroniveusin B I de I to idea

2 ’,3’-Dihydroniveusin A I del to idea Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. S : !

t 1 11 1 i i i I 1 1 1! ! i 1 n

s à f ^ J i I I ?

Sx X 55 5 S S 5 5

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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. U l

g £

11- I

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Substituent Name of Compound Plant Source Ref

3,4/9-Epoxyelemasteiroct inol ide Cri Ionia qucdrongular

11a, 13-01 hydrogongrothcninol ide

7a-Hydroxygcngrothamnolide Gongrothomnue aurontiCO Substituent Name of Compound Plant Source

H H /î-We.H 1 la. 13-Oihydroelemanol id< Critonia quadrangular Is 126

OH OI-Val-2-OH CHg Schkuhrldin A Schkuhrla schkuhrIoldes 57

OH OH CH, Schkuhrldin B Schkuhrla schkuhrlaides 57

H f-Me.H 1 la,13-01hydroelemastelraclI no IIde Critonia quadrangulc

OH CHg 15-Hydroxyelemaslelroctlnol Ide Stevio achalensis

OH a-Me,H Ilf,13-01hydro-15-hydroxy Stevla achalensis eIemostelroctInolI de Plant Source SubstituentStructure Name of Compound

8/î-Acetoxyreynosin Hydropectis oquotica

7a-Hydroxyreyno3În Decochoelo ovatifolio

Sonchuside D Sonchus oleroceus

tt-OAc 1-Epiludoibin Mikonio guoco 43

a-CMoc Ba-Methocry1oy1oxybo1chon i n Chomoemelum fuscolum

a-Oi-Bul 6a-Isobutyryloxybalchanin Chomoemelum fuscotum

|9-€Mac-4-OH Lontonifoline Zexmenio ionfonifolio

7a-Hyd roxyaont omarine Decochoeto ovotifolio

9/î-+lydroxytournefort loi ide

9/î-Aceloxytournefort iol ide

8a-Acetoxyarbusculin B Substituent Name of Compound Plant Source

8a-Methacryloyloxyarmexi fol if Chomaemelum fuscalur

Ba-laobutyry(oxyortnexifol in Chomoemelum fuscolun

3^-lsobulyryloxyortecalin Greenmanielto r

î/î-lsobulyryloxy-4-epiartecai ir Greenmoniello r Il

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Substituent Name of Compound Plant Source Ref

8a-f 4-Hydr oxyme t hocryIoyIoxy]- Onopordon carmanicum 215 4-epi-sonchucorpolide

8a-[4-HydroxyiTiethacryloy loxy]- Onopordon carmanicum 215 sonchucorpolIde

8a-Acetoxy-3a-hydroxyarbusculir Mikanio guaco 43

Ba-Methacryloyloxyarmefclin Chmaemelum fuscotum 53

8a-lsobutyryloxyarinefol in Chomoemelum fuscotum 53

*No stereochemistry i Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Substituent Name of Compound

a-OH Me Isotosolide Laser trilobum

*/J-OAc H Isosi lerol ide Lcserpitium si le

•Known compound stereochemistry r

9-Oxo-tournefort iolide Artemisia tournefort ic

Eudesmon-4,11-dien-12,8/?-ol ide Artemisia iwoyomogi 92 Substituent Name of Compound Plant Source

3a,5a-0ihydroxyisoaIanto lactone Artemisia i woyomog i

3a-Peroxy i soo1 onto 1actone Artemisia iwoyomogi

3a-Peroxy-5a-hydroxy i soo1 onto 1actone Artemisia iwoyomogi 92

f-CH 3a-Peroxy-5/!-hydroxy i soo 1 an to 1 actone Artemisia iwoyomogi 92

3a-Hydroxyeude3ma-4,11-die 12,8jS-ol ide

RO 3a-Peroxyeudesmc-4,11-dien- 12,8j?-ol ide

4a-Per oxy-eudesmo-2,11-die 12,8f-olide I I

I I

f'3 f-!

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. u

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. t i f

U I t s I Il ti ël.T i, m 1,1 fi - U m = = E ! i s i I I I

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Il I I

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. is

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. P--

)■s >- A ^;

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Name of Compound Plant Source

8/}-5'[5* '-Dihydroxyligloyl]-4 Corphochoete bigelovii 160 hydroxytlgIoyIoxydesocet y I- zuubergenin

8/f-T igloyl oxypreeupor t undin Eupolorium olliaaimum 33 Tig ,5'-0ihydroxyligloyloxy]- Eupotorlum oltissimum 33 prceupotundln

3/?-Acetoxy-1a-hydroxy-8o-f&- Gutenbergio morginota 127 occloxyongeIoyIoxyj-eremon thin

3^-^cetoxy-1a-hydroxy-8a-[4,5- Gutenbergio morginota 127 diaceloxyongeloyloxyj-erefnonthin 11

* i sri Ssi Ss|

5 § I

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.1.

Is Is 11

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. g <5

ill

ttXXXXOÔÔÔ

. . È kxxxo585SÔ

xo 8x5555

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I If I « ' II1 4 I t It I t l i

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. H i l II lï

Bejaranoa balansae

2^,8^-Dihydroxyguoia-3,10(14),11(13)- Hymenothrix wisiizenii 122 tr ien-12,6a-olI de

OH OH Tig f 1: Bejaranoa balansae

5-Deaoxy-8-deacyIeuparoti n-8-O- Eupatorium altlssimun 33 [4',5'-dihydroxyt iglatej

Bejaranoa balansae 223 Substi tuent Name of Compound Plant Source

I O T ig

BalansolIde Bejaranoa balansae / H = Ov,

H % OAC CebeIMn H Centaurea bello RO— \ 1 R2 OMac-4-OH Cebel(in G Centaureo beIIa

HO''^ H = Diaspanoside C Diaspananthus unifloru:, 4 R1 O'

0 X

Cl Ang C«2 DesocyichlorojanerIn B-O-ongelate Gutenbergia marginata H \ Ang fWe.H Oesacy1-11,13-dihydrGchIorojoner in Gutenbergio marginata

M0C-4-CH Cebel1 in C HO J 0R1 CH2 Centaurea adjorica G. Cebel1 in D R-nA hV CH2 Centaureo adjarica HO 0- Mac CHg Repdiolide tr iol Centaurea incana OC(CHj)-CHCH20H 11 II I ? III iii

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Plant Source Ref Substituent Name of Compound

i-Sut la: Artemisia douglasionc1 123

i-Val Artemisia douglasionc

Mebut 1c: Artemisia douglasionc

Tig Artemisia douglasionc

11^,13-Oihydro-epi-ligustrin Eriocepholus giessii

i-eut a-Me.H Artemisia douglosiono

a-Me,H Artemisia douglosiono

\ Mebut a-Me.H Artemisia douglosiono \''0 )""0R a-Me.H Artemisia douglosiono

a-Me.H Artemisia douglosiono ' o Y ... CH2 Artemisia douglosiono 0 Substituent Name of Compound Plant Source Ref

Ixeria dentoto

12-Methoxydihydrodehydro Sousaureo lappa

coatualactone

CH2 Subexpinnotin C Centaurea canar ienaia

M0C-4-CH CH2 Subexpinnatin B Centaurea canarienaia

Mac-4-OH CH2 Clementein B Centaurea clementei

Mac-4-OH ^-H,a-We •Clemenlein C Centaurea clementei

Me Mac-4-OH CH2 ♦Clementein Centaureo clementei

•Known compounda now atereochemiatry oaaigned ot C-15 O > > > >

k 4 2 •« 1 I I î II li {■! îi

Il li il A 8- A & -Î5& ,55^II

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! ? î

1 1 1

- s s i î i

I S I I XI

W «r I I < <

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5 5

B «

fiiil = = X i I I

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. iî

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. structure Substituent Name of Compound Plant Source

Tonnunolide B Tanacetum annuum

'O R M= H Tannunolide A Tanocetum annuum

Bedfordiolide Bedfordia orborescens

Iso-seco-tanaparlholide Substituent Name of Compound Plant Source

«No stereochemistry is given

Ambrosia moritima OH CH2 lOa-Hydroxydomsin

11a,13-epoxide 2,3-Dihydrostromonin B Ambrosia moritima

Desocy1 chi op in A-ocetate For theniurn lozonionum C«2

CH2 Desocy1 chiopin A-isovolerale For theniurn lozonianum

Ambrosia confer!I flora 248 Substituent Name of Compound

CoronopiIin-15-O-isovolerate Partheniurn lozonionum 132

Coronopil in-1&-0-[2-fnethyl butyrole] Porthenium lozonionum 132

Isochiopine B Porthenium fruticosum 186

14-Acetoxytetroneurin D Porthenium lozonionum 132

15-Oesocetyltetroneurin C Porthenium lozonionum 132

Desocetyltetroneurin D-15-0- Porthenium lozonionum 132 isobutyrote

15-Oesocetyltetroneurin C- Porthenium lozonionum 132 iscbutyrote

Desocetyltetroneurin D-4-0- Porthenium lozonionum 132 isobutyrote

Porthenium frutic 8 8 8

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. i I

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. I I •8 8

I? 1 i It i ti }f II

Vertiojolcanol Ide B-O-ocetate Vernonia jalcono

Vernonio jalcono

I-But 13-O-Methylvernojaicanolide 8-0- Vernonia jalcono ''0R1 isobutyrafe 13-O-Melhylvernojolcanolide 8-0- Vernonio jalcono methacrylole

Arteonnuin C Artemisio annuo

8a-Tigloyloxy-vernomorgolide Vernonio morginolo Ch-f~ OTig 0 i i

11 .? I T o Ô It-

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. structure Substituent Name of Compound plant Source

3: Gochnat i a poIymorpho

4: Gochnotio polymorpho

Gochnotio polymorpho

H 8: Gochnotio polymorphe

9: Gochnotio polymorphe

Gochnotio polymorphe

Gochnot io polymorpho f-H 11:

Gochnotio polymorphe

Versicolactons C Aristolochio versicolo

hexonoyI Thapsia gargonict

Octanoyl Thopsia garganic< Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.3. Common side chains in sesquiterpene lactones

Structure of Elemental Type of Abbrevi a t ion Side Chain Composition Ester

C2f% Ethyl

C~H,0 Acetate

C3H5O Propyl

C4H5O M e th a c ry la te Mac

4-Hydroxymethacrylate Moc-4-CH

C4H5O2 EpoxymethocryI a te Epo

Q I C^HgOCI 2 -H y d ro x y -3 -c h lo ro - A«

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.3. (continued)

Structure of Elemental Type of S id e Chain Compas it ion Ester

0C(CHj)=CHCH20H

Isobutyrcte

0 OH

C4H7O2 2,3-Oihydroxybutyrote But-2,3-OH

C4H7O2 2,3-Oihydroxy- isobutyrote

CgHyO Ange I ate

C5H7O Senec i oote

C^HyO Tig la te T ig

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.3. (continued)

Structure of ElementalElemento Abbrevlot ion Side Cha Compos i t ion 0

OH 4-Hydroxysenecioote Sen-OH

C^Hy0 2 4-Hydroxytiglote Tig-4-OH

C^HyOg 5-HydroxyangeIote Ang-5-OH

Epoxyongclote Epoxyong

OH C5H7O2

C5 H7 O2 4 .5-Dihydroxytiglate Tig-4,5-OH

CcHgO 2-MethyIbutyrote Mebut

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.3. (continued)

Structure of Elementol Type of Composition Ester Side Chain

XX C5H9O

CgHgOg 2-HydroxyisovoIerote i-Vcl-2-OH

C5H9O4

CgHnO Hexonoy I

CfiH,.0.- Glucose

CyHgOj (2o-Acetoxyethyl) E« ocrylote

CyHgOj 4-Acetyltiglote Tig-4-A

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.3. (continued)

structure of Elementol Type of Side Choin Composition Ester .OAc

Cy-I^O^ 4-Acety longe lote Ang-4-Ac

CyHgOj Acetyisorrocinote Soroc

OAc

0 OH

OAc Cy-igO^ 4-Acetoxy-S- hydroxytiglote OH

° , ^OAc ^7^11^3 4-Acetoxysenecioote Sen-Ac

(XCh^ \ V___ OH CgHyOg 4-Hydroxyphenylocetyl

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.3. (continued)

Structure of Elemental Type of Abbreviotion Side Choi Composition Ester 4-fXethoxybenzcyl B2l(4-OMe)

CgH^jO Octanoyl

CgHg02 4-MethoxybenzyI JJ»

^9^11^5 <.5-Oiocetoxyongelote Ang-4,5-Ac

^9^11°5 4.5-D!ocetoxyt iglote Tig-4.5-Ac

CgH^jOg 2,3-DiocetQxy-2- mebut-2,3-Ac

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.3. (continued)

Composi tie

Ctt/^isOs

P P CnHl5°4 "O

HO' R= 1 ).....OH

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.3. (continued)

Structure of Elemental Abbrevlot It Side Chain Composition

CiiH^gOg R. h o '

Cl2'^15°6

R = COCH

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table 1.3. (continued)

Elemental Abbrevlot!(

HO 'o r R = COCH=CH

HO — f V CH=CHCOOCH^

CZ5M35O4

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. REFERENCES

1. Abdel Salam, N. A.; Mahmoud, Zeinab F.; Kassem, Fahima K. Egypt. J. Pharm. Sci., 27(1-4), 275-82, 1986.

2. Abdullaev, N. D.; Nurmukhamedova, M. R. , Khim. Prir. Soedin., 2,240-2, 1987.

3. Adegawa, Shigeru; Miyase, Toshio; Fukushima, Seigo, Chem. Pharm. Bull., 34(9), 3769-73, 1986.

4. Adegawa, Shigeru; Miyase, Toshio; Ueno, Akira, Chem. Pharm. Bull., 35(4), 1479-85, 1987.

5. Adekenov, S. M.; Abdykalykov, M. A.; Turdybekov, K. M.; Kagarlitskii, A. D.; Gatilov, Yu. V., Izv. Akad. Nauk Kaz. SSR Ser. Khim., (5), 74-9, 1986.

6. Adekenov, S. M.; Aituganov, K. A.; Golovtsov, N. I., Khim. Prir. Soedin., (1), 148-9, 1987.

7. Adekenov, S. M.; Gafurov, N. M.; Turmukhambetov, A. Zh.; Ivlev, V. I., Khim. Prir. Soedin., (2). 305-6, 1987.

8. Adekenov, S. M.; Kharasov, R. M.; Kupriyanov, A. N.; Turmukhambetov, A. Zh., Khim. Prir. Soedin., (5), 644-5, 1986.

9. Adekenov, S. M.; Kupriyanov, A. N. ; Kagarlitskii, A. D. USSR, Vestn. Akad. Nauk Kaz. SSR, (10), 52- 62, 1986.

10. Aguirre-Galviz, L. E. , Fitoterapia, 58(1), 50-1, 1987.

11. Ahmad, V. U.; Zamir, T.; Hasan, N. M. ; Albert, K., J. Chem. Soc. Pak., 8(3), 425-8, 1986.

12. Ahmad, Viqar Uddin; Fizza, Kaniz, Liebigs Ann. Chem., (7), 643-4, 1987.

13. Ahmad, Viqar Uddin; Fizza, Kaniz, Phytochemistry, 25(4), 949-50, 1986.

14. Ahmed, Ahmed A.; Whittemore, Alan T. ; Mabry, Torn J. , J. Nat. Prod., 49(2), 362-3, 1986.

15. Aleskerova, A. N.; Serkerov, S. V., Izv. Akad. Nauk Az. SSR, Ser. Biol. Nauk, (3), 43-5, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 16. Aleskerova, A. N.; Serkerov, S. V. , Izv. Akad. Nauk Az. SSR, Ser. Biol. Nauk, (4), 28-30, 1986.

17. Alonso-Lopez, Manuel; Borges-del- Castillo, Juan; Rodriguez-Ubis, Juan C.; Vazquez-Bueno, Purificacion, J. Chem. Sac., Perkin Trans. 1, (12), 2017-19, 1986.

18. Ananthasubramanian, L.; Gopinath, H. ; Bhattacharyya, S. C., Indian J. Chem., Sect. B, 25B(4), 380-3, 1986.

19. Ando, Masayoshi; Wada, Tetsuo; Kusaka, Haruhiko; Takase, Kahei; Hirata, Naonori; Yanagi, Yoshikazu, J. Org. Chem., 52(21), 4792- X6, 1987.

20. Appendino, Giovanni; Calleri, Mariano; Chiari, Giacomo; Gariboldi, Pierluigi; Menichini, Francesco, Gazz. Chim. Ital., 116(11), 637-41, 1986.

21. Appendino, Giovanni; Gariboldi, Pierluigi; Belliardo, Flavio, Phytochemistry, 25(9), 2163-5, 1986.

22. Appendino, Giovanni; Nano, Gian Mario; Calleri, Mariano; Chiari, Giacomo, Gazz. Chim. Ital., 116(2), 57-61, 1986.

23. Appendino, Giovanni; Valle, Maria Grazia; Caniato, Rosamaria; Cappelletti, Elsa Maria, Phytochemistry, 25(7), 1747-9, 1986.

24. Appendino, Giovanni; Valle, Maria Grazia; Gariboldi, Pierluigi, J. Chem. Soc., Perkin Trans. 1, (8), 1363-72, 1986.

25. Appendino, Giovanni; Valle, Maria Grazia; Gariboldi, Pierluigi, Phytochemistry, 26(6), 1755-7, 1987.

26. Banerjee, S. ; Schmeda-Hirschmann, G.; Castro, V. ; Schuster, A.; Jakupovic, J.; Bohlmann, F., Planta Ned., (1), 29-32, 1986.

27. Barbetti, Paolo; Casinovi, Carlo Giulio, Ann. 1st. Super. Sanita, 1981, 17(2), 255-82.

28. Barbetti, Paolo; Casinovi, Carlo Giulio, Ann. 1st. Super. Sanita. 1981, 17(2), 283-322.

29. Barrero, Alejandro F.; Sanchez, Juan F. ; Zafra, Ma Jose; Barron, A.; San Feliciano, Arturo, Phytochemistry, 26(5), 1531-3, 1987.

30. Benesova, V., Rastit. Resur. , 22(1), 134-42, 1986.

31. Benezra, Claude; Epstein, William L. , Contact Dermatitis, 15(4), 223-30, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 32. Boeker, R.; Jakupovic, J.; Bohlmann, F.; Schmeda- Hirschmann, G., Planta Med., 53(1), 105-6, 1987.

33. Boeker, Ruth; Jakupovic, Jasmin; Bohlmann, Ferdinand; King, Robert M.; Robinson, Harold, Phytochemistry, 25(7), 1669-72, 1986.

34. Bohlmann, Ferdinand; Zdero, Christa; Jakupovic, Jasmin; Grenz, Michael; Castro, Victor; King, Robert M.; Robinson, Harold; Vicent, Leszek P. D., Phytochemistry, 25(5), 1151-9, 1986.

35. Bohlmann, Ferdinand, Nat. Prod. Chem., Proc. Int. Symp. Pak.-U.S. Binatl. Workshop, 1st, Meeting Date 1984, 65-89. Edited by: Rahman, Atta Ur. Springer: Berlin, Fed. Rep. Ger., 1986.

36. Bohlmann, Ferdinand; Zdero, Christa; King, Robert M.; Robinson, Harold, Liebigs Ann . Chem., (5), 799-813, 1986.

37. Bohlmann, Ferdinand; Zdero, Christa; Schmeda-Hirschmann, Guillermo; Jakupovic, Jasmin; Dominguez, Xorge A.; King, Robert M.; Robinson, Harold, Phytochemistry, 25(5), 1175-8, 1986.

38. Bordoloi, M. J.; Sharma, R. P.; Sarma, J. C., Tetrahedron Lett., 27(38), 4633-4, 1986.

39. Calderon, J. S.; Quijano, L.; Gomez, F.; Rios, T., J. Nat. Prod. B50(3), 522, 1987.

40. Calderon, Jose S.; Quijano, Leovigildo; Gomez-Garibay, Federico; Sanchez, Donato M.; Rios, Tirso; Fronczek, Frank R., Phytochemistry, 26(6), 1747-50, 1987.

41. Cappelletti, Elsa M.; Caniato, Rosamaria; Appendino, Giovanni, Biochem. Syst. Ecol., 14(2), 183-90, 1986.

42. Carda, Miguel; Arno, Manuel; Marco, J. Alberto, Tetrahedron, 42(13), 3655-62, 1986.

43. Castro, V.; Jakupovic, J.; Bohlmann, F., Phytochemistry, 25(7), 1750-2, 1986.

44. Catalan, Cesar A. N.; De Iglesias, Dora I. A.; Kavka, Juan; Sosa, Virginia E.; Herz, Werner, J . Nat. Prod., 49(2), 351-3, 1986.

45. Chandra, Amitabh; Misra, Laxmi N.; Thakur, Raghunath S. Phytochemistry, 26(5), 1463-5, 1987.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 46. Chiari, Giacomo; Appendino, Giovanni; Nano, Gian Mario, J. Chem. Soc., Perkin Trans. 2, (2), 263-6, 1986.

47. Ciminiello, Patrizia; Fattorusso, Ernesto; Magno, Silvana; Mayol, Luciano, Can. J . Chem., 65(3), 518-22, 1987.

48. Collado, I. Gonzalez; Macias, F. A.; Massanet, G. M.; Luis, F. Rodriguez, Tetrahedron, 42(13), 3611-22, 1986.

49. Collado, Isidro G.; Macias, Francisco A.; Massanet, Guillermo M.; Molinillo, Jose Maria G.; Rodriguez- Luis, Francisco, J. Org. Chem., 52(15), 3323-6, 1987.

50. D’Ambrosio, Michele; Guerriero, Antonio; Pietra, Francesco Helv. Chim. Acta, 70(3), 612-20, 1987.

51. Daniewski, Wlodzimierz M.; Kroszczynski, Wojciech; Bloszyk, Elzbieta; Drozdz, Bohdan; Nawrot, Jan; Rychlewska, Urzula; Budesinsky, Milos; Holub, Miroslav, Collect. Czech. Chem. Commun., 51(8), 1710-21, 1986.

52. De Luengo, Doris H.; Miski, Mahmut; Gage, Douglas A.; Mabry, Tom J., Phytochemistry, 25(8), 1917-22, 1986.

53. De Pascual Teresa, J.; Caballero, E. ; Anaya, J. ; Caballero, C.; Gonzalez, M. S., Phytochemistry, 25(6), 1365-9, 1986.

54. De Pascual Teresa, Joaquin; Moran, Joaquin R. ; Hernandez, Jose M.; Grande, Manuel, Phytochemistry, 25(5), 1167-70, 1986.

55. De la Fuente, Rosa J.; Diaz, Oscar I.; Morales, Isabel; Oberti, Juan C.; Sosa, Virginia E.; Herz, Werner, Planta Ned., (3), 243, 1986.

56. Delgado, Guillermo; Alvarez, Laura; Huerta, Eduardo; Romo de Vivar, Alfonso; Mata, Rachel, Hagn. Reson. Chem., 25(3), 201- 2, 1987.

57. Delgado, Guillermo; Guzman, Salvador; Romo de Vivar, Alfonso, Phytochemistry, 26(3), 755-9, 1987.

58. Dhillon, R. S.; Kalsi, P. S.; Singh, W. P.; Gautam, V. K.; Chhabra, B. R., Phytochemistry, 26(4), 1209-10, 1987.

59. Dominguez X. A.,Cano G., Sanchez H., Velazquez G., J. Nat. Prod.,49(4), 704-5, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 60. Dzhafarov, A. I.; Mir-Babaev, N. F.; Sereda, N. P., Izv. Akad. Nauk Az. SSR. Ser. Biol. Nauk, (2), 100-2, 1986.

61. El Dahmy, S.; Sarg, T.; Salem, S. A.; Jakupovic, J.; Bohlmann, P., Planta Med., (5), 365-7, 1986.

62. El Sebakhy, Nadia A.; El Ghazouly, Maged G. , Pharmazie, 41(4), . 298, 1986.

63. El-Emary, Nasr A.; Makboul, Makboul A.; Hamed, Mahmound Phytochemistry, 26(1), 314-15, 1986.

64. El-Ghazouly, Maged; Omar, Abdallah A.; El-Din, Ahmed Self; Khafagy, Saad M., Acta Pharm. Jugosl. , 37(2), 131-3, 1987.

65. El-Sebakhy, Nadia A.; El-Ghazouly, M. G. ; El-Din, A. A. Seif; Zdero, C., Pharmazie, 41(7), 525-6986, 1986.

66. Elissalde, Marcel H., Jr.; Ivie, G. Wayne, Am. J. Vet. Res., 48(1), 148-52, 1987.

67. Ellmauerer, E.; Pathak, V. P.; Jakupovic, J.; Bohlmann, F.; Dominguez, X. A.; King, R. M.; Robinson, H. Phytochemistry, 26(1), 159-63, 1986.

68. Emerenciano, Vicente De P.; Kaplan, Maria Auxiliadora C.; Gottlieb, Otto R.; Bonfanti, Maria Renata De M.; Ferreira, Zenaide S.; Comegno, Leonora M. A., Biochem . Syst. Ecol., 14(6), 585-9, 1986.

69. Emerenciano, Vicente de P.; Kaplan, Maria Auxiliadora C.; Gottlieb, Otto R., Biochem . Syst. Ecol., 13(2), 145-66, 1985.

70. Estrada, M. D.; Conde, A.; Marquez, R.; Jimenez Garay, R. Acta Crystallogr., Sect . C: Cryst. Struct. Commun., 042(10), 1413-15, 1986.

71. Falsone, Gioacchino; Haddad, Hatim; Wendisch, Detlef Arch. Pharm. (Weinheim, Ger.), 319(4), 372-9, 1986.

72. Fang, Nianbai; De Luengo, Doris H.; Mabry, Tom J. Phytochemistry, 25(11), 2665-6, 1986.

73. Fernandez, Isabel; Garcia, Begona; Grancha, Francesc J.; Pedro, Jose R. , Phytochemistry, 26(8), 2403-5, 1987.

74. Fraga, B. M. Natural Products Reports, 273-96, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 75. Fujimoto, Yoshinori; Kinoshita, Takumi; Ikekawa, Nobuo; Mungarulire, Joseph, Phytochemistry, 26(9), 2593-5, 1987.

76. Galvez, M.; Hoeneisen, M.; Silva, M.; Watson, W. H. Bol. Soc. Chil. Quim. . 31(1), 3-5, 1986.

77. Gao, Feng: Mabry, Tom J., Phytochemistry, 25(1), 137-40, 1986.

78. Gao, Feng; Miski, Mahmut; Mabry, Tom J. , Phytochemistry, 25(5), 1231-3, 1986.

79. Gao, Feng; Wang, Huiping; Mabry, Tom J., J. Nat. Prod., 50(1), 23-9, 1987.

80. Gao, Feng; Wang, Huiping; Mabry, Tom J. , Phytochemistry, 26(3), 779-81, 1987.

81. Gaspar, A. R. M. D.; Potgieter, D. J. J.; Vermeulen, N. M. J., Biochem. Pharmacol ., 35(3), 493-7, 1986.

82. Gaspar, A. R. M. D.; Verschoor, J. A.; Neitz, A. W. H. ; Vermeulen, N. M. J. , Biochem. Pharmacol., 36(15), 2461-5, 1987.

83. Gennari, M.; Abbattista Gentile, I.; Cugudda, L. Z. Pflanzenkrankh. Pflanzenschutz, 94(1), 68-73, 1987.

84. Gil, Roberto R.; Oberti, Juan C. ; Sosa, Virginia E.; Herz, Werner, Phytochemistry, 26(5), 1459-61, 1987.

85. Gogte, M. G.; Ananthasubramanian, L.; Nargund, K. S.; Bhattacharyya, S. C., Indian J. Chem., Sect. B, 25B(3), 233-8, 1986.

86. Goldsby, Gwendolyn; Burke, Basil A., Phytochemistry, 26(4), 1059-63, 1987.

87. Gonzalez Collado, I.; Macias, F. A.; Massanet, G. M.; Rodriguez Luis, F., Rev. Latinoam . Quim., 16(4), 128-41, 1986.

88. Gonzalez, A. G., Actual. Chim. Ther., 1984, 11, 3-37.

89. Gonzalez, Antonio; Galindo, Antonio; Falenzuela, Jose A.; Mansilla, Horacio, Tetrahedron Lett., 27(24), 2771-4, 1986.

90. Goren, Nezhun; Ulubelen, Ayhan, Phytochemistry, 26(9), 2585-7, 1987.

91. Goren, Nezhun; Ulubelen, Ayhan, Phytochemistry, 26(9), 2633-4, 1987.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 92. Greger, Harald; Zdero, Christa; Bohlmann, Ferdinand Phytochemistry, 25(4), 891-7, 1986.

93. Groenewegen, W. A.; Knight, D. W.; Heptinstall, S. , J. Pharm. Pharmacol., 38(9), 709- 12, 1986.

94. Guerreiro, Eduardo, Phytochemistry, 25(3), 748-50, 1986.

95. Guerrero, C.; Nava, A. L.; Obregon, L., Rev. Latinoam. Quim., 18(2), 57-8, 1987.

96. Gutierrez, Alicia B.; Oberti, Juan Carlos; Sosa, Virginia E. ; Herz, Werner, Phytochemistry, 26(8) 2315-20, 1987.

97. Hall, Iris H.; Lee, K. H.; Sykes, Holly C., Planta Ned., 53(2), 153-6, 1987.

98. Haruna, Mitsumasa; Sakakibara, Yoshihisa; I to, Kazuo, Chem. Pharm. Bull., 34(12), 5157-60, 1986.

99. Hayashi, Toshimitsu; Koyama, Junko; McPhail, Andrew T.; Lee, Kuo Hsiung, Phytochemistry, 26(4), 1065-8, 1987.

100. He, Linxing; Zhang, Jiu; Xue, Huizhong, Zhiwu Xuebao, 29(2), 197-203, 1987.

101. Herz, Werner, Nat. Prod. Chem., Proc. Int. Symp. Pak.-U.S. Binatl. Workshop, 1st, Meeting Date 1984, 154-74. Edited by: Rahman, Atta Ur. Springer: Berlin, Fed. Rep. Ger. 1986.

102. Herz, Werner; Bruno, Maurizio, Phytochemistry, 25(8), 1913-16, 1986.

103. Herz, Werner; Bruno, Maurizio, Phytochemistry, 26(1), 201-4,

104. Herz, Werner; Sosa, Virginia E., Phytochemistry, 25(6), 1481-3, 1986.

105. Hoeneisen, Maritza; Becker, Hans, J. Nat. Prod., 49(2), 359-60,

106. Hoffmann, H. M. R.; Rabe, Juergen, Angew. Chem., 97(2), 96-112, 1985.

107. Holub, Miroslav, Int. Conf. Chem. Biotechnol. Biol. Act. Nat. Prod., [Proc.], 1st, Volume 2, 83- 95. Edited by: Atanasova, B. Bulg. Acad. Sci.: Sofia, Bulg. 1981.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 108. Holub, Miroslav, Stud. Org. Chem. (Amsterdam), 20 (Nat. Prod. Chem.), 1985, 37-50.

109. Holub, Miroslav; Budesinsky, Milos, Phytochemistry, 25(9), 2015-26, 1986.

110. Holub, Miroslav; Budesinsky, Milos; Smitalova, Zdenka; Saman, David; Rychlewska, Urszula, Collect . Czech. Chem. Commin. , 51(4), 903-29, 1986.

111. Holub, Miroslav; Toman, Jan; Herout, Vlastimil, Biochem. Syst. Ecol., 15(3), 321-6, 1987.

112. Hubert, Terrance D.; Okunade, Adewole L.; Wiemer, David F., Phytochemistry, 26(6), 1751-3, 1987.

113. Huneck, Siegfried; Zdero, Christa; Bohlmann, Ferdinand Phytochemistry, 25(4), 883-9, 1986.

114. Hutchinson, John H.; Money, Thomas; Piper, Susan E. , Can. J. Chem., 64(7), 1404-6, 1986.

115. Ishitsuka, Midori; Kusumi, Takenori; Kakisawa, Hiroshi; Kawakami, Yoshiyuki; Nagai, Yasushi; Sato, Tadashi, Tetrahedron Lett., 27(23), 2639-42, 1986.

116. Itoigawa, Masataka; Takeya, Kazumi; Furukawa, Hiroshi; I to, Kazuo, J. Cardiovasc . Pharmacol., 9(2), 193-201, 1987.

117. Itokawa, Hideji; Matsumoto, Hajime; Mizuno, Kenji; Watanabe, Kinzo; Morita, Hiroshi; Iitaka, Yoichi, Chem. Pharm. Bull., 34(11), 4682-6, 1986.

118. Jacobs, Helen; Bunbury, Malcolm; Hylands, Peter J.; McLean, Stewart, J. Nat . Prod., 49(6), 1163-4, 1986.

119. Jakupovic, J.; Banerjee, S.; Castro, V.; Bohlmann, F.; Schuste, A.; Msonthe, J. D.; Keeley, S., Phytochemistry 25(6), 1359-64, 1986.

120. Jakupovic, J.; Chen, A.-L.; Bohlmann, F., Phytochemistry, 26(10), 2777-9, 1987.

121. Jakupovic, J.; Jia Y.; Zdero. C.; Warning, U.; Bohlmann, F.; Jones, S. B., Phytochemistry, 26(5), 1467-9, 1987.

122. Jakupovic, J.; Schuster, A.; Bohlmann, F.; Sanchez, H. V.; Dominguez, X. A., Phytochemistry, 26(9), 2543-6, 1987.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 123. Jakupovic, J.; Chau-Thi, T. V.; Warning, U., Bohlmann, F.; Greger, H., Phytochemistry, 25(7), 1663-7, 1986.

124. Jakupovic, J.; Banerjee, S.; Castro, V.; Bohlmann, F.; Schuster, A.; Msonthi, J. D.; Keeley, S., Phytochemistry, 25(6), 1359-64, 1986.

125. Jakupovic, J.; Baruah, R. N.; Bohlmann, F.; King, R. M.; Robinson, H., Planta Med ., (3), 204-6, 1986.

126. Jakupovic, J.; Baruah, R. N.; Bohlmann, F.; Msonthi, J. D. Planta Med., (2), 154-5, 1986.

127. Jakupovic, J.; Boeker, R.; Schuster, A.; Bohlmann, F.; Jones, S. B., Phytochemistry. 26(4), 1069-75, 1987.

128. Jakupovic, J.; Castro, V.; Bohlmann, F., Phytochemistry, 26(2), 451-5, 1987.

129. Jakupovic, J.; Castro, V.; Bohlmann, F., Phytochemistry, 26(7), 2011-17, 1987.

130. Jakupovic, J.; Gage, D. A.; Bohlmann, F.; Mabry, T. J. Phytochemistry. 25(5), 1179-83, 1986.

131. Jakupovic, J.; Jia, Y.; Pathak, V. P.; Bohlmann, F.; King, R. M., Planta Med.. (5), 399-401, 1986.

132. Jakupovic, J.; Pathak, V. P.; Bohlmann, F.; Dominguez, X. A., Phytochemistry, 26(3), 761-4, 1987.

133. Jakupovic, J.; Pathak, V. P.; Bohlmann, F.; Gage, D.; Dillon, M. 0., Phytochemistry, 25(11), 2563-5, 1986.

134. Jakupovic, J.; Pathak, V. P.; Bohlmann, F. ; King, R. M.; Robinson, H., Planta Med ., (4), 331-2, 1986.

135. Jakupovic, J.; Schmeda-Hirschmann, G.; Schuster, A. ; Zdero, C.; Bohlmann, F.; King, R. M.; Robinson, H. ; Pickardt, J., Phytochemistry, 25(1), 145-58, 1986.

136. Jakupovic, J.; Schuster, A.; Bohlmann, F.; Sanchez V., H.; Del Rio, S., E. ; Dominguez, X. A., Rev. Latinoam. Quim., 17(3- 4), 210-11, 1986.

137. Jakupovic, J.; Sun, H.; Bohlmann, F.; King, R. M., Planta Med., 53(1), 97-8, 1987.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 138. Jakupovic, J.; Sun, H.; Geerts, S. ; Bohlmann, F., Planta Med., 53(1), 49-51, 1987.

139. Jakupovic, Jasmin; Jia, Yunyi; King, Robert M.; Bohlmann, Ferdinand, Liebigs Ann. Chem., (8), 1474-7, 1986.

140. Jakupovic, Jasmin; Zdero, Christa; Boeker, Ruth; Warning, Ulrich; Bohlmann, Ferdinand; Jones, Samuel B., Liebigs Ann. Chem., (2), 111-23, 1987.

141. Kagarlitskii, A. D.; Adekenov, S. M., Vestn. Akad. Nauk Kaz. SSR, 1984, (10), 9-17.

142. Kasymov, Sh. Z., Khim. Prir. Soedin., (5), 551-69, 1982.

143. Kasymov, Sh. Z.; Ovezdurdyev, A.; Yusupov, M. I.; Sham’yanov, I. D.; Malikov, V. M., Khim . Prir. Soedin., (5), 636, 1986.

144. Kelsey, Rick G.; Locken, Laura J., J. Chem. Ecol., 13(1), 19-33, 1987.

145. Kuroda, Miyuki; Yoshida, Daisuke; Kodama, Hisashi, Agric. Biol. Chem., 51(2), 585-7, 1987.

146. Kutschabsky, L.; Kretschmer, R. G.; Ripperger, H., Cryst. Res. Technol., 21(5), 627-33, 1986.

147. Lange, Gordon L.; Lee, Moses, J. Org. Chem., 52(3), 325-31, 1987.

148. Lange, Gordon L.; Lee, Moses, Synthesis, (2), 117-20, 1986.

149. Mahajan, R.; Singh, Prabhsaran; Bajaj, K. L.; Kalsi, P. S. Nematologica, 32(1), 119- 23, 1986.

150. Mahmoud, Z. F.; Kassem, F. F. ; Abdel- Salam, N. A.; Zdero, C., Phytochemistry, 25(3), 747-8, 1986.

151. Mahmoud, Zainab F.; Abdel Salam, Nabil A., Egypt. J. Pharm. Sci., 27(1-4), 271-4, 1986.

152. Mahmoud, Zainab F.; Kasem, Fahima F.; Abdel Salam, Nabil A., Egypt. J. Pharm. Sci., 27(1-4), 283-9, 1986.

153. Malcolm, Arcelio J.; Fischer, Nikolaus H., J. Nat. Prod., 50(2), 167-72, 1987.

154. Maldonado, Emma; Flores, Adrian M.; Ortega, Alfredo J. Nat. Prod., 49(6), 1152, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 155. Maldonado, Emma; Mayorga, Lilia; Ortega, Alfredo Phytochemistry, 26(1), 205-7, 1986.

156. Martinez, Mariano; Esquivel, Baldomero; Ortega, Alfredo, Phytochemistry, 26(7), 2104-6, 1987.

157. Massiot, Georges; Morfaux, Anne Marie; Le Men-Olivier, Louisette; Bouquant, James; Madaci, Abdelaziz; Mahamoud, Abdallah; Chopova, Mariette; Aclinou, Paul, Phytochemistry, 25(1), 258-61, 1986.

158. Matusch, Rudolf; Haeberlein, Hanns, Liebigs Ann. Chem., (5), 455-7, 1987.

159. Metwally, M. A.; El-Dahmy, S.; Jakupovic, J.; Bohlmann, F.; Dawidar, A. M. ; Metwally, S. A., Phytochemistry, 25(1), 255-7, 1986.

160. Meurer, Barbara; Seaman, Fred C.; Mabry, Tom J., Phytochemistry, 26 (6), 1743-6, 1987.

161. Mir-Babaev, N. F.; Serkerov, S. V., Khim. Prir. Soedin., (1), 110-11, 1986.

162. Miski, Mahmut; De Luengo Doris H.; Mabry, Tom J. Phytochemistry, 26(1), 199-200, 1986.

163. Miski, Mahmut; Mabry, Tom J., Phytochemistry, 25(7), 1673-5, 1986.

164. Misra, Laxmi N., Phytochemistry, 25(12), 2892-3, 1986.

165. Miyase T.; Fukushima S., Chem . Pharm. Bull., 35, 2869, 1987.

166. Miyase, Toshio; Yamada, Mayumi; Fukushima, Seigo, Chem. Pharm. Bull., 35(5), 1969-74, 1987.

167. Moiseeva, G. P.; Yusupova, I. M.; Abduazimov, B. Kh.; Sidyakin, G. P., Khim. Prir . Soedin., (2), 176-9, 1986.

168. Morales, Glauco; Borquez R., Jorge; Mancilla P., Arlett; Pedreros T., Samuel; Loyola M., Luis A. , Phytochemistry, 25(10), 2412-14, 1986.

169. Morales, Glauco; Borquez, Jorge; Mancilla, Arlett; Pedreros, Samuel; Loyola, Luis A., J . Nat. Prod., 49(6), 1140-1, 1986.

170. Nagumo, Seiji, Hoshi Yakka Daigaku Kiyo, (28), 1-8, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 3 0 0 171. Nawrot, Jan; Bloszyk, Elizbieta; Harmatha, JuraJ; Novotny, Ladislav; Drozdz, Bohdan, Acta Entomol. Bohemoslov., 83(5), 327-35, 1 plate, 1986.

172. Nishimura, Keiichi; Miyase, Toshio; Ueno, Akira; Noro, Tadataka; Kuroyanagi, Masanori; Fukushima, Seigo Phytochemistry, 25(10), 2375-9, 1986.

173. Nishimura, Keiichi; Miyase, Toshio; Ueno, Akira; Noro, Tadataka; Kuroyanagi, Masanori; Fukushima, Seigo, Chem. Pharm. Bull., 34(6), 2518-21, 1986.

174. Nowak, Gerard; Drozdz, Bohdan; Holub, Miroslav, Acta Soc. Bot. Pol., 55(2), 233-8, 1986.

175. Nowak, Gerard; Drozdz, Bohdan; Holub, Miroslav; Budesinsky, Milos; Saman, David, Acta Soc. Bot. Pol., 55(2), 227-31, 1986.

176. Nowak, Gerard; Drozdz, Bohdan; Holub, Miroslav; Lagodzinska, Agnieszka, Acta Soc . Bot. Pol., 55(4), 629-37, 1986.

177. Nowak, Gerard; Drozdz, Bohdan; Kroszczynski, Wojciech; Holub, Miroslav, Acta Soc . Bot. Pol., 55(1), 17-22, 1986.

178. Ober, Alfonso G.; Fischer, Nikolaus H. , Phytochemistry, 26(3), 848-9, 1987.

179. Ober, Alfonso G.; Fischer, Nikolaus H.; Parodi, Felix Phytochemistry, 25(4), 877-81, 1986.

180. Ober, Alfonso G.; Urbatsch, Lowell E.; Fischer, Nikolaus H., Phytochemistry, 25(2), 467-70, 1986.

181. Ober, Alfonso G.; Urbatsch, Lowell E.; Fischer, Nikolaus H., Phytochemistry, 26(4), 1204-6, 1987.

182. Oberti, Juan C.; Gil, Roberto R. ; Sosa, Virginia E.; Herz, Werner, Phytochemistry, 25(6), 1479-80, 1986.

183. Oberti, Juan C.; Silva, Gloria L.; Sosa, Virginia E.; Kulanthaivel, Palaniappan; Herz, Werner, Phytochemistry, 25(6), 1355-8, 1986.

184. Oksuz, Sevil; Ayyildiz, Hatice, Phytochemistry, 25(2), 535-7, 1986.

185. Okuda, Takuo; Yoshida, Takashi; Chen, Xin Min; Xie, Jing Xi; Fukushima, Makoto, Chem. Pharm. Bull., 35(1), 182-7, 1987.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 186. Ortega, Alfredo; Maldonado, Emma, Phytochemistry, 25(3), 699-701, 1986.

187. Qshima, Yoshiteru; Wong, Siu Ming; Konno, Chohachi; a Cordell, Geoffrey A.; Waller, Donald P.; Soejarto, D. Doel; Fong, Harry H. S., J. Nat. Prod., 49(2), 313-17, 1986.

188. Ovezdurdyev, A.; Abdullaev, N. D. ; Kasymov, Sh. Z.; Akyev, B., Khlm. Prir. Soedin., (5), 568-71, 1986.

189. Pathak, V. P.; Khanna, R. N., Phytochemistry, 26(7), 2103-4, 1987.

190. Pearce, John; Gershenzon, Jonathan; Mabry, Tom J. Phytochemistry, 25(1), 159-65, 1986.

191. Perez, Ana L.; Nava, Leticia; Romo de Vivar, Alfonso Phytochemistry, 25(3), 745-6, 1986.

192. Pfeil, Richard M.; Gage, Douglas A.; Lee, Esther P.; Miski, Mahmut; Mabry, Tom J.; Powell, A. Michael Phytochemistry, 26(1), 195-8, 1986.

193. Pieman, A. K.; Arnason, J. T.; Lambert, J. D. H., J. Nat. Prod., 49(3), 556, 1986.

194. Pieman, Anna K., Biochem . Syst. Ecol., 14(3), 255-81, 1986.

195. Pieman, Anna K., Biochem . Syst. Ecol., 15(3), 361-3, 1987.

196. Pirrung, Michael C.; Thomson, Stephen A., Tetrahedron Lett., 27(24), 2703-6, 1986.

197. Purushothaman, K. K.; Vasanth, Saradha, Indian J. Chem., Sect. B, 25B(4), 417-18, 1986.

198. Quijano, Leovigildo; Calderon, Jose S. ; Gomez-Garibay, Federico; Angulo-M., Margarita; Rios, Tirso, Phytochemistry, 26(9), 2589-92, 1987.

199. Quijano, Leovigildo; Calderon, Jose S. ; Gomez-Garibay, Federico; Bautista, Simeon; Rios, Tirso; Fronczek, Frank R., Phytochemistry, 25(3), 695-7, 1986.

200. Quinkert, Gerhard; Schmalz, Hans Guenther; Walzer, Egon; Kowalczyk-Przewloka, Teresa; Duerner, Gerd; Bats, Jan W., Angew. Chem., 99(1), 82-4, 1987.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 201. Rigby, James H.; Senanayake, Chrisantha, J. Am. Chem. Soc.. 109(10), 3147-9, 1987.

202. Rigby, James H.; Wilson, Jo Ann Zbur, J. Org. Chem., 52(1), 34-44, 1987.

203. Rojatkar, Supada R.; Dhaneswar, Narayandatta, N.; Puranik, Vedavati G.; Tavale, Sudam S.; Gururow, Tayur N. ; Nagasampagi, Bhimsen A., J. Chem. Research (M), 2543-64, 1986.

204. Romo de Vivar, Alfonso, Ciencia (Mexico City), 32(3), 163-89, 1981.

205. Romo de Vivar, Alfonso; Delgado, Guillermo, Bol. Soc. Chil. Quim., 30(2), 79-100, 1985.

206. Romo de Vivar, Alfonso; Delgado, Guillermo; Soriano- Garcia, Manuel; Toscano, Ruben A.; Huerta, Eduardo; Reza-Garduno, Rosa G., J. Nat. Prod ., 50(2), 284-6, 1987.

207. Rosik, G. G. ; Zinchenko, A. A.; Reznichenko, A. A.; Kovalev, I. P., Khim.-Farm . Zh., 21(5), 632-4, 1987.

208. Rossi, Carlo; Ambrogi, Valeria; Grandolini, Giuliano; Scarcia, Vito; Furlani, Ariella, J . Pharm. Sci., 75(8), 784-6, 1986.

209. Rossiter, Marycarol; Gershenzon, Jonathan; Mabry, Thomas J., J. Chem. Ecol., 12(6), 1505-21, 1986.

210. Rustaiyan, A.; Ganji, M. T., Phytochemistry, 26(10), 2857-9, 1987.

211. Rustaiyan, A.; Ahmadi, B.; Jakupovic, J.; Bohlmann, P., Phytochemistry, 25, 1659, 1986.

212. Rustaiyan, A.; Bamonieri, A. ; Raffatrad, M.; Jakupovic, J.; Bohlmann F., Phytochemistry, 26(8), 2307-10, 1987.

213. Rustaiyan, A.; Jakupovic, J.; Chau- Thi, T. V.; Bohlmann, F. ; Sadjadi, A., Phytochemistry, 26(9), 2603-6, 1987.

214. Rustaiyan, Abdolhossein; Sharif, Zahra; Tajarodi, Azade; Sadjadi, Abulfazl S., Phytochemistry, 26(10), 2856-7, 1987.

215. Rustaiyan, A.; Ahmadi, B.; Jakupovic, J.; Bohlmann, F. Phytochemistry, 25(7), 1659-62, 1986.

216. Rustaiyan, A.; Dabiri, M.; Jakupovic, J., Phytochemistry, 25(5), 1229-30, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 217. Rychlewska, Urszula; Hodgson, Derek J.; Grabarczyk, Kalina; Drozdz, Bohdan; Daniewski, Wlodzimierz M.; Kroszczynski, Wojciech; Budesinsky, Milos; Holub, Miroslav, Collect. Czech. Chem. Comman., 51(8), 1698-709, 1986.

218. Saitbaeva, I. M.; Abdullaev, N. D.; Mallabaev, A.; Sidyakin, G. P.; Yagudaev, M. R., Khlm . Prir. Soedin., (1), 115-16, 1986.

219. San Feliciano, Arturo; Medarde, Manuel; Poza, Maria Teresa; Miguel Del Corral, Jose M., Phytochemistry, 25(7), 1757-9, 1986.

220. Santarius, K. A.; Falsone, G.; Haddad, H., Toxicon, 25(4), 389-99, 1987.

221. Sarma, Nittala S.; Rambabu, M.; Anjaneyulu, A. S. R.; Rao, C. B. S., Indian J. Chem., Sect. B, 26B(2), 189-90, 1987.

222. Schmeda-Hirschmann, G.; Boeker, R.; Jakupovic, J.; Bohlmann, F. , Phytochemistry, 25(7), 1753-4, 1986.

223. Schmeda-Hirschmann, G.; Jakupovic, J.; Pathak, V. P.; Bohlmann, F., Phytochemistry, 25(9), 2167-70, 1986.

224. Schmeda-Hirschmann, G.; Zdero, C.; Bohlmann, F. Phytochemistry, 25(7), 1755-6, 1986.

225. Schultz, Arthur G.; Motyka, Linda A.; Plummer, Mark, J. Am. Chem. Soc., 108(5), 1056-64, 1986.

226. Seaman, Fred C.; Fischer, Nikolaus H.; Mabry, I. J. Phytochemistry, 25(11), 2663-4, 1986.

227. Seaman, Fred C.; Funk, V. A., Taxon, 32(1), 1-27, 1983.

228. Seaman, Frederick C., Dot . Rev., 48(2), 121-595, 1982.

229. Segal, R.; Feuerstein, I.; Danin, A., Biochem. Syst. Ecol., 15(4), 411-16, 1987.

230. Serkerov, S. V.; Aleskerova, A. N., Khim. Prir. Soedin., (1), 101-4, 1987.

231. Serkerov, S. V.; Aleskerova, A. N., Khim. Prir. Soedin., (5), 645-7, 1986.

232. Seto, Mamoru; Miyase, Toshio; Fukushima, Seigo, Chem. Pharm. Bull., 34(10) 4170-6, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 233. Shibuya, Hirotaka; Ohashi, Kazuyoshi; Kawashima, Keiko; Hori, Kazuyuki; Murakami, Nobutoshi; Kitagawa, Isao, Chem. Lett., (1), 85-6, 1986.

234. Smitalova, Zdenka; Budesinsky, Milos; Saman, David; Holub, Miroslav, Collect. Czech. Chem. Commun., 51(6), 1323-39, 1986.

235. Smitt, Ulla Wagner; Moldt, Peter; Christensen, Soeren Broegger, Acta Chem. Scand., Ser. B, B40(9), 711- 14, 1986.

236. Spring, Otmar; Klemt, Volker; Albert, Klaus; Hager, Achim, Z. Naturforsch., C : Biosci., 41(7-8), 695-8, 1986.

237. Spring, Otmar; Priester, Theodor; Hager, Achim, J. Plant Physiol., 123(1), 79-89, 1986.

238. Stefanovic, Milutin; Aljancic-Solaja, Ivana; MilosavlJevic, Slobodan, Phytochemistry, 26(3), 850-2, 1987.

239. Stevens, Kenneth L., Isopentenoids Plants: Biochem. Funct., [Pap.-Symp.], Meeting Date 1982, 65- 80. Edited by: Nes, W. David; Fuller, Glenn; Tsai, Lee-Shin. Dekker: New York, N. Y. 1984.

240. Stevens, Kenneth L.; Wong, Rosalind Y. , J. Nat. Prod., 49(5), 833-7, 1986.

241. Stierle, Donald B., Phytochemistry, 25(3), 743-5, 1986.

242. Ta1war, K. K.; Kalsi, P. S., Phytochemistry, 25(1), 262-3, 1986.

243. Tashkhodzhaev, B.; Karimov, Z. ; Yusupov, M. I., Khim. Prir. Soedin., (3), 291-5, 1986.

244. Thastrup, O.; Linnebjerg, H.; BJerrum, P. J.; Knudsen, J. B. ; Christensen, S. Broegger, Biochim . Biophys. Acta, 927(1), 65-73, 1987.

245. Ulubelen, A.; Oksoz, S.; Goren, N., New Trends in Natural Products Chemistry 1986. Studies in Organic Chemistry, Vol. 26. 513-28. Edited by: Atta-ur Rahman, and P. W. Le Quesne 1986.

246. Ulubelen, Ayhan; Goren, Nezhun, J. Nat. Prod., 49(6), 1104- 5, 1986.

247. Ulubelen, Ayhan; Goren, Nezhun; Jakupovic, Jasmin Phytochemistry, 26(1), 312-13, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 248. Vargas, David; Fronczek, Frank R.; Fischer, Nikolaus H.; Hostettmann, Kurt, J. Nat. Prod., 49(1), 133-8, 1986.

249. Warning, U.; Jakupovic, J.; Bohlmann, F.; Jones, S. B., Liebigs Ann. Chem., (5), 467-8, 1987.

250. Washino, Tsutomu; Kobayashi, Hidetoshi; Ikawa, Yoshitomi, Agric. Biol. Chem., 51(6), 1475-80, 1987.

251. Watanabe, Keisuke; Oguri, Yukio; Miyakado, Masakazu J. Nat. Prod., 49(2), 362-3, 1986.

252. Watanabe, Keisuke; Oguri, Yukio; Miyakado, Masakazu Planta Med. , (2), 112-14, 1986.

253. Watanabe, Keisuke; Ohno, Nobuo; Mabry, Tom J., Phytochemistry, 25(1), 141-3, 1986.

254. Woerdenbag, H. J., Pharm . Weekbl., Sci. Ed., 8(5), 245-51, 1986.

255. Woerdenbag, H. J.; Meijer, C.; Mulder, N. H. ; De Vries, E. G. E.; Hendriks, H. ; Malingre, T. M., Planta Med., (2), 112-14, 1986.

256. Wu, Jin Bin; Chun, Yui To; Ebizuka, Yutaka; Sankawa, Ushio, Chem. Pharm. Bull., 1985, 33(9), 4091-4.

257. Yusupova, I.M.; Tashkhodzhaev, B. ; Mallabaev, A., Khim. Prir. Soedin., 6,788-90, 1986.

258. Zdero, C.; Bohlmann, F., Phytochemistry, 26(9), 2597-2601, 1987.

259. Zdero, C.; Bohlmann, F.; Muller, M., Phytochemistry, 26(10), 2763-75, 1987.

260. Zdero, C.; Bohlmann, F., Planta Med., 53(2), 169-72, 1987.

261. Zdero, C.; Bohlmann, F.; King, R. M.; Robinson, H. Phytochemistry, 26(4), 1207-9, 1987.

262. Zdero, C.; Bohlmann, F.; King, R. M.; Robinson, H., Planta Med. , (1), 22-4, 1986.

263. Zdero, C.; Bohlmann, F.; Schmeda- Hirschmann, G. Phytochemistry, 26(2), 463-6, 1987.

264. Zdero, C.; Bohlmann, F.; Scott, R., Phytochemistry, 26(7), 1999-2006, 1987.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 265. Zhang, Decheng; Haruna, Mitsumasa; McPhail, Andrew T.; Lee, Kuo Hsiung, Phytochemistry, 25(4), 899-904, 1986.

266. Zhang, Jiu; He, Linxing, Yaoxue Xuebaa, 21(4), 273-8, 1986.

267. Zheng, Jie; Li, Genpei; Chen, Zhongguo; Tang, Youqi; Wei, Xincheng; He, Linxing, Huaxue Xuebaa, 44(6), 551-7, 1986.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix A

NATURAL SOURCE VERSUS REFERENCE

Acanthospermam hispidum

Achillea micrantha

AJanla fastlgiata 244

Ambrosia artemisilfolia 239

Ambrosia confertiflora 249

Ambrosia maritima 138

Ambrosia tenuifolia 184

Anvil lea garcini 217

Arctotis arctotoides

Aristolochia versicolar

Artemisia annua

Artemisia anomal a

Artemisia argentea

Artemisia austriaca 6

Artemisia caerulescens 220

Artemisia douglasiana 123

Artemisia fraga 231

Artemisia iwayomogi

Artemisia leucodes

Artemisia maritima 190

Artemisia nitrosa 8

Artemisia rutifolia 113

Artemisia szowitsiana 232

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix A (continued) Natural source versus reference

Artemisia tournefortiana 213

Artemisia xanthochroa 113

Austroliabum candidum 125

Ayapana el at a 128

Bartlettina karwinskiana 78

Bedfordia arborescens 262

Bejaranoa balansae 224

Blainvillea latifolia 204

Brachylaena elliptica 259

Calea jamaicensis 9

Calea megacephala

Calea rupicola

Calea septuplinervia

Calea zacatechichi

Carphochaete bigelovii 160

Centaurea adjarica 176

Centaurea bel la 176

Centaurea calcitrapa 131

Centaurea canariensis 87

Centaurea clementei 48, 87

Centaurea collina 74

Centaurea incana 157

Centaurea repens 131, 241

Centaurea uni flora 21

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix A (continued) Natural source versus reference

Centratherum punctatum 26, 140

Chaenactis douglasii 242

Chamaemelum fuscatum 53

Coriaria Japonica 186

Cotula cinerea 159

Critonia quadrangular is 128

Critonlopsis huaircajana 119

Decachaeta ovatifolia

Decachaeta scabrella 162

Diaspananthus uniflorus 4

Dittrichia gravec lens 214

Dittrichia viscosa

Elephantopus angustifolius

Elephantopus mollis 26,121

Elephantopus scaber 266

Elephantopus tomentosus 99

Erigeron khorassanicus 189

Eriocephalus africanus 260

Eriocephalus giessii 260

Eriocephalus kingessii 260

Eriocephalus scariosus

Eupatorium altissimun

Eupatorium cannabium

Eupatorium fortunei

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix A (continued) Natural source versus reference

Eupatorium quadrangulare 112

Ferula communis 163

Ferula penninervis 2

Gaillardia megapotamica 134

Gaillardia powellii 103

Gochnatia foliolosa 105

Gochnatia hypoleuca 37

Gochnatia polymorpha 37

Gongrothamnus aurantiaca 140

Greenmaniella resinosa

Gutenbergia cordiflora 75

Gutenbergia marginata 127

Helenium autumnale 158

Helenium donianum 263

Helianthus heterophyllus 79, 102

Helianthus microcephallus 79

Helianthus niveus 254

Helianthus resinosus 191

Helogyne hutchisonii 133

Hydropectis aquatica 136

Hymenothrix wislizenii 122

Inula caspica 5

Inula graveolens 20

Isocarpha oppositifolia 192

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix A (continued) Natural source versus reference

Ixeris dentata 233

Jurinella moschus 211

Lactuca laciniata 173

Lactuca sativa 150

Laser trilobum 235

Laserpitium garganicam 23

Laserpitium slier 110

Hikania cordiflora 96

Nlkania guaco

Hikania micrantha

Hikania vitifolia

Hiileria quinqueflora 129

Hontanoa frutescens 200

Hontanoa gigas 199

Hontanoa imbricata

Hontanoa leucantha 188

Huschleria stolzii 140

Neohintonia monantha 72

Onopordon carmanicum 212

Parthenium fruticosum 187

Parthenium lozanianum 132

Perityle vaseyi 193

Perymenium discolor 155

Perymenium mendezii 155

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix A (continued) Natural source versus reference

Picris hieracioides 174

Polymnia maculata 153

Prenanthes acerifolia 167

Pseudoelephantopus spicatus 135

Ratibida columnifera 67

Rudbeckia laciniata 139

Saussurea lappa

Schkuhria schkuhrioides

Senecio caudatus 34

Senecio rosmarinus 169

Smyrnium connatum 91

Smyrnium creticum 248

Smyrnium galaticum 247

Smyrnium olusatrum

Smyrnium perfoliatum 90

Sonchus oleraceus

Spbaeranthus indicus 85

Spirastrella inconstans 222

Stevia achalensis 36

Stevia amambayensis 225

Stevia aristata 264

Stevia breviaristata 183

Stevia mercedensis 36

Stevia ovafa 197

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix A (continued) Natural source versus reference

stevia origanoides 39

Stevia protrerensis 84

Tanacetum annuum 29

Tanacetum santolino ides 65

Tanacetum vulgare 45

Thapsia garganica 71, 236

Tithonia rotundifolia 17

Torilis Japonica 117

Urospermum dalechampii 218

Vernonia beliinghanii 140

Vernonia brachycalyx 140

Vernonia chioropappa 140

Vernonia cinerea 119

Vernonia cistifolia 140

Vernonia condensate 140

Vernonia cotoneaster 135

Vernonia diffusa 140

Vernonia erdverbengii 59

Vernonia holstii 140

Vernonia Jalcana 135

Vernonia marginata 130

Vernonia mollissima 44

Vernonia patens 135

Vernonia poskeanolide 119

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix A (continued) Natural source versus reference

Vernonia scorpioides 250

Vicoa indica 198

yjguiera deltoldea 77

Viguiera gilliesii 94

Viguiera ladibractate 80

Wedelia pinetorum 104

Youngia denticulata 3

Zexmenia lantanifolia 95

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B Sesquiterpene Lactones Reported in 1986 and 1987

COMPOUND NAME VERSUS REFERENCE

8a-Acetoxy-10a-hydroxy-l, 13-bis-O-methyl 135 hirsutinolide

8a-Acetoxy-1Oa-hydroxy-13-0-me thy1 135 hirsutinolide

8a-Acetoxy-10a-hydroxyhirsutinolide 135

15-Acetoxy— 9a-hydroxy— 8g-methacryloyl 129 oxy-14-oxo-acanthospermolide

15-Acetoxy-1-hydroxy-6a-1 igloy1oxy-1 Oa- 104 methyl-ToH,8oH,lipH-eudesm-4-olide

8a-Acetoxy-llB,13-dihydrogluco 258 zaluzanin C

8a-Acetoxy-lip,13-dihydroparishin A 259

3B-Acetoxy-llp,13-epoxyep itulipinolide 78

14-Acetoxy-11«13-dihydrodesoxyivangus t in 259

3p-Acetoxy-la-hydroxy-8a-[4,5-diacetoxy 127 angeloyloxy]-eremanthin

3B-Acetoxy-la-hydroxy-8a-[5-acetoxy 127 angeloyloxy]-eremanthin

8p-Acetoxy-2a-hydroxycostunolide 260

8a-Acetoxy-3a-hydroxyarbusculin B 43

3a,4a-epoxide 8a-Acetoxy-4a,5a-epoxy 135 J a1cagua i ano1ide 13-0-acetate

3a,4a-epoxide 8a-Acetoxy-4a,5a-epoxy- 135 13-0-methyl jalcaguaianolide

8g-[4' -Acetoxy-5’ -hydroxytigloyloxy]-novanin 133

14-Acetoxy-5a-hydroperoxy-lla,13- 259 dihydroisoalantolactone

14-Acetoxy-5a-hydroperoxy-isoalantolactone 259

316

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

3p-Acetoxy-8p-5’[S'’-hydroxytigloyl]-4’ hydroxytigloyloxycostunolide

15-Acetoxy-8p-[2-methylbutyryloxy]-14- oxo-germacra-l(10)E,4E-dien-12, 6a-olide

3p-Acetoxy-8p-tigloyloxyheliangolide 133

2a-Acetoxy-8-desacylglaucolide-E-acetate 140

lp-Acetoxy-8-hydroxyeudesma-3, 7(11)- 246 dien-8 ,12-olide

2g-Acetoxy-8a,13-dihydroxygermacra-l (10), 4,7(ll)-trien-6a,12-olide

3p-Acetoxy-8a-[2,3-diacetoxy-2- methylbutyryloxy]-eremanthin

3p-Acetoxy-8a-[3-acetoxy-2-hydroxy-2- methylbutyryloxy]-eremanthin

3/3-Acetoxy-8a- [4, 5-diacetoxyangeloyl oxy]-eremanthin

3p-Acetoxy-8a-[4-acetoxyangeloyloxy]- eremanthin

3p-Acetoxy-8a-[5-acetoxyangeloyloxy]- eremanthin

13-Acetoxy-8a-hydroxy-7,11-dehydro- 11, 13-dihydroanhydroverlotorin

15-Acetoxy-9a-methacryloyloxy-8p- hydroxy-14-oxo-acanthospermolide

lg-Acetoxy-eudesma-3,7(11),8-trien- 8 ,12-olide

ip-Acetoxy-eudesma-4(15),7(11),8-trien- 8 ,12-olide

8a-Acetoxyabusculin B

15-Acetoxydentatin B

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

14-Acetoxydesacetyl laurenobiolide 155

Sp-Acetoxyrnosin 136

14-Acetoxytetraneurin D 132

8j3-[4-Acetoxytigloyloxy]-14-oxo-4Z- 224 acanthospermolide

8p-[4-Acetoxytigloyloxy]-14-0X0- 224 acanthospermo1ide

9g-Acetoxytournef ort iolide 212

8-Acetyl-13-ethoxypiptocarphol 44

2-Acetyl-8|3-[4,5-dihydroxytigloyloxy]- 260

4-Acetyl-8-epi-inuviscolide 196

3-Acetylchamissonin 77

Ajafinin 243

Altamisic acid 183

8a-Angeloyloxy-1Oa-hydroxy-1-desoxy 135 hirsutinolide

3a,4a-epoxide 8a-Angeloyloxy-4a,5a- 135 epoxyjalcaguaianolide

3a,4a-epoxide 8a-Angeloyloxy-4a,5a- 135 epoxyj a1cagua i ano1ide 13-0-acetate

3a,4a-epoxide 8a-Angeloyloxy-4a,5a- 135 epoxy-13-0-me thy1 j a1cagua i anolide

2p-Angeloyloxy-8a-(2’-methyl)butyryl 234 oxy-1Op-ace toxy-11a-hydroxyslov-3-eno1ide

8a-Angeloyloxycompactifloride 135

8a-Angeloyloxyhirsutinolide-13-0-acetate 135

Argophyllin C 253

Artanomaloide 120

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

Artausin 6

Arteannuin C 164

Artecaninhydrate 113

Artegallin 219

Artelin 218

Artesovin 231

Balansolide 223

Bedfordiolide 261

Ba,15-Bis(acetoxy)anhydroverlotorin 140

8,13-Bis-deacylvernonataloide-8-0-tiglate 130

1,11-Bis-epi-artesin 92

4,8-Bis-epi-inuviscolide 261

8 ,10-Bis-epi-mikanokryptin 134

Bis-seco-tanapartholide 113

Brachycalyxolide 140

Brachynereolide 258

Breviarolide 182

Britanin 5

Calcine E 156

Calcine F 156

14-Carboxy-15-demcthy1costunolide 26

15-Carboxy-15-desme thy1-11(13)- 26 dchydrosaussurea lactone

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

Cebellin B 175

Cebellin C 175

Cbellin D 175

Cebellin E 175

Cebellin F 175

Cebellin G 175

Cebellin H 175

Cebellin I 175

Chlororepdiolide 240

Cistiglaucolide-8 -O-methacrylate 140

Clementein 87

Clementein B 87

Clementein C 87

Cnicin-4’-0-acetate 131

Confertiflorin 248

Corianin 185

Coronopilin-15-G-[2-methyl butyrate] 132

Coronopilin-15-Ü-isovalerate 132

Deacetoxy-3-oxo-chromolaenide 33

Deacetoxy-3-oxo-eupaformin tiglate 33

8-Deacetoxylaserolide 234

Deacetyl chromolaenide 33

Deacetyl eupaformin-8-0-[4’,5’-dihydroxy 33 tiglate]

Deacetyl eupaformin-8 -0 -[4’-hydroxytiglate] 33

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

Deacetyl eupaformin-8 -O-tiglate 33

Deacetyl tulipinolide-lp,lOa-epoxide 160

14-Deacetyl-17, 18-dehydrorotundin 17

Deacetylargentolide B 63

13-Deacetylmarginatin 130

8-Deacylvernonataloide-8-0-tiglate 130

6-epi-l,lO-Dehydro-10,14-dihydro 160 chrysostamolide acetate

11.13-Dehydromelitensin-diacetate 140

10.14-Dehydrostilpnotomentolide-8-0-tiglate 59

2-epi-Deoxyorthopapp-4E-enolide 121

9-Desacetoxy-14-0-(4-hydroxysenecioyl) 140 vernocistifolide-8 -Q-angelate

9-Desacetoxy-14-0-senecioyl 140 vernocistifolide-8 -O-angelate

10-Desacetoxy-9p-hydroxyglaucolide A 140

2-Desacetoxygaillardin-p-D- 262 glucopyranoside tetraacetate

9-Desacetoxymelcanthin F 5

Desacetyl linifolin B 134

Desacetylacanthospermal A 129

15-Desacetyltetraneurin C 132

15-Desacetyltetraneurin C-isobutyrate 132

Desacetyltetraneurin D-15-O-isobutyrate 132

Desacetyltetraneurin D-4-Q-isobutyrate 132

Desacyl chiapin A-acetate 132

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

Desacyl chiapin A-isovalerate 132

Desacyl-11,13-dihydrochlorojanerin 127

8 -Desacylcentratherin-8-0-[2,3-epoxy 140

isobutyrate]

Desacylchlorojanerin 8-0-angelate 127

8 -Desacylglaucolide A-tiglate 59

Desacyl grazielic acid acetate 84

Desacylgrazielic acid-[4-acetoxytiglate] 224

Desacylisoelephantopin senecioate 121

Desacylisoelephantopin tiglate 121

Desacylisovalerylheliangine 181

8 -Desacylmarginatin 8 -0-acetate 135

Desacyltagitinin C-8-0-[2-methylbutyrate] 264

Desacyltagitinin E-8 -O-propionate 264

Desacyltagitinin F-8-0-[2-methylbutyrate] 264

8-Desacylvernonataloide 8-0-acetate 135

8 -Desacylvernonataloide 8-0-angelate 135

8 -Desacylvernonataloide 8-0-isobutyrate 135

1-Desoxy-8-(2,3-epoxyisobutyryloxy)-10a- 140 hydroxyhirsutinolide

5-Desoxy-8-deacyleuparotin-8-0- 33 [4’,5’-dihydroxytiglate]

2-Desoxyflorilenalin-p-D- 262 glucopyranoside tetraacetate

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

Desoxytagitinin B-3-O-methyl ether 264

8a,13-Di ace toxy-1a-hydroxygerma era- 160 4E,7(ll),9Z-trien-6a,12-olide

1.15-Diacetoxy-6a-tigloyloxy-10a-methyl- 104 7aH, BaH,llpH-eudesm-4-en-B,12-olide

1.15-Diacetoxy-6a-tigloyloxy-10a-methyl- 104 7aH,BaH-eudesma-4,11-diene-B,12-olide

8a-13-Diacetoxy-7,11-dehydro-11, 13- 160 dihydroanhydroverlotorin

Ba,lla-Diangeloyloxy-lOg- 23 acetoxyslov-3-enolide

Diaspanolide A 4

Diaspanolide B 4

Diaspanoside A 4

Diaspanoside B 4

Diaspanoside C 4

4.15-Didehydro-4,5-dihydro-la-hydroxy 36 steiractinolide

IP, 10a; 4a, 5/3-Diepoxy-8/3- 90 isobutoxyglechomanolide

Ip,10a;4a,Sp-Diepoxy-Ba- 90 isobutoxyglechomanolide

lip,13-Dihydro-l-epi-inuviscolide 213

10(14)-Dihydro-1op-methoxy-1-epi- 136 inuviscolide

10(14)-Dihydro-10a-methoxy-inuviscolide 136

lip,13-Dihydro-15-hydroxy 36 elemasteiractinolide

17-Dihydro-17-hydroxy-18-chloro 119 vernonataloide

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

lljS, l-Dihydro-19-hydroxyvernolide

1ip,13-D ihydro-1a-hydroxy-8 -ep i- xerantholide

4P,15-Dihydro-3p-acetoxy-8p-5’[S''- hydroxytigloyl]-4’-hydroxytigloyloxy

4P,15-Dihydro-3p-acetoxy-8p-5’[S’’-acetoxy tigloyl]4’-hydroxytigloyloxyzaluzanin C

8p-(2,S-Dihydro-S-hydroxy-3-furylcarbontyl oxy)-2 -desoxodehydroleucodin

4a,lS-Dihydro-7a-hydroxy-3- desoxyzaluzanin C

lip,13-Dihydro-7a-hydroxy-8-epi- xerantholide

4p,lS-Dihydro-8p-S’[S’’-acetoxytigloyl]- 4 ’-hydroxytigloyloxyzaluzanin C

lip,13-Dihydro-epi-ligustrin

lip,13-Dihydroarctiolide

16,17-Dihydrobrachycalyxolide

lip,13-Dihydrodeoxyelephantopin 26S

11a,13-Dihydroelemaasteiractinolide 128

11a,13-Dihydroelemanolide 128

lip,13-Dihydroglucozaluzanin C 258

11a,13-Dihydrogongrothamnolide

lip,13-Dihydroivangustin acetate 259

2’,3’-Dihydroniveusin A 77

2’,3’-Dihydroniveusin B

2’,3’-Dihydroniveusin C

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

2,3-Dihydrostramonin B 138

113.13-Dihydrovernolide 140

11a,13-Dihydroxerantholide 161

4a,lOa-Dihydroxy-1,5H-guaia-2,11(13)- 259 dien-12,6a-olide

33-14-Dihydroxy-113,13-dihydro-costunolide 150

53,lOa-Dihydroxy-laH-dehydroleucodin 113

13,4a-Dihydroxy-2a,3a-epoxy- 90 83-methoxy-eudesma-7(1 1 ),8 -en-8a,12-olide

13,4a-Dihydroxy-2a,3a-epoxy- 90 eudesma-7(ll),8-dien-8 ,12-olide

43,10a-Dihydroxy-3-oxo-83“ 264 isobutyryloxyguaia-11(13)-en-12,6a-olide

4a,10a-Dihydroxy-3-oxo-83- 264 isobutyryloxyguaia-11(13)-en-12,6a-olide

2a,14-Dihydroxy-83-(2’R,3’R)-2’ ,3’- 190 epoxyangeloyloxyco stunolide

33,93-Dihydroxy-83-tigloxycostunolide 223

4a,lOa-Dihydroxy-8 -acetoxy-la,5a,113H- 159 guaia-2 -en-1 2 ,6a-olide

14,15-Dihydroxy-9a-acetoxy-83- 127 isobutyryloxyacanthospermo1ide

8a,93-Dihydroxy-trans,trans-germacra- 45 l(10),4 -dien-trans-6 ,12-olide

113.13-Dihydroxyepi tulipinolide 78

113,53-Dihydroxyeriocephaloide 259

2 3 ,83-Dihydroxyguaia-3,10(14),11(13)- 122 trien-12,6a-olide

3a,5a-Dihydroxyisoalantolactone 92

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

8p-5’[S''-Dihydroxytigloyl]-4’-hydroxy 160 tigloyloxydesacetylzuubergenin

Sp-[4’ ,S'-Dihydroxytigloyloxy]- 33 preeupatundin

4,S-Dioxo-lipH-xanth-l(10)-en-12,Sp-olide 213

Douglasine 241

(E)-lS-Acetoxydentatin-A-acetate 140

(E)-8a,IS-Diacetoxy-l-oxo-p-cyclocostunolide 140

[1(10)E,4E]-Germacra-1(10).4-dien- 249 12,8a-olide-lS-oic acid

(E,E)-8a,lS-Diacetoxy-14-oxo-l(10), 140 4,1(13)-germacratrien-12, 6 -olide

4E-Deacetyl chromolaenide-4'-0-acetate 33

1-Epiludalbin 43

4P,IS-Epoxy-ip,Sp-oxidomiller-9E-enolide 129

lip, 13-Epoxy-ll,13-dihydroaromatin 134

14,10a-Epoxy-4-hydroxyglechoma-8-enolide 247

u p , 13-Epoxy-8p-acetoxy-a-cyclocostunolide 78

4P,lS-Epoxy-8 -desacyImi1ler-9E- 129 enolide-8 -O-angelate

Ip,1Op-Epoxyd ihydropar theno1i de 214

3,4p-Epoxyelemasteiractinolide 128

lip,13-Epoxyepitulipinolide 78

la,10p-Epoxygermacra-4E,11(13)- 128 dien-1 2 ,8a-olide

4P,lS-Epoxymiller-1(10)2,8E-dienolide 129

4P,lS-Epoxymiller-9E-enolide 129

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

4^,15-Epoxymiller-9Z-enolide 129

la. 10|3-Epoxyovatifolin 264

Ergolide 188

ip-Ethoxy-4p,15-epoxymiller-9Z-enolide 129

15-Ethoxy-4oH-isocentratherin 26

1^-Ethoxymiller-9Z-enolide 129

Eucannabinolide diacetate 133

Eucannabinolide-19-O-acetate 191

Eudesman-4,ll-dien-12,8^-olide 92

Eupafortunin 98

Eupatoriopicrin 19-0-1inolenoate 260

Fercolide 163

Ferulide 2

trans-trans Germacra-1(10),4- 105 dien-cis-6 ,12olide

Gigantanolide A 198

Gigantanolide B 198

Gigantanolide C 198

Graveolide 20

Grazielic acid-[4-hydroxytiglate] 263

Guaia-4(15),10(14),ll(13)-trien-12,8p-olide 36

Gutenbergin 75

llaH,13-Dihydroconfertin 51

llpH,13-Dihydrourospermal A 217

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

8a-[Hydroxymethacryloyloxy]- 215 sonchucarpolide

8a-[Hydroxymethacryloyloxy]- 215 4-epi-sonchucarpolide

14-Hydroxy-desoxyivangustin 259

Helenalin angelate 262

lip,13-Hydro-8-epi-confertin 213

lip,13-Hydrobrachynereolide 258

4a-Hydroperoxy-10a-hydroxy-la,5aH-guaia- 259 2,ll(13)-dien-12,6a-olide

4a-Hydroperoxy-10a-hydroxy-8a-acetoxy- 259 la,5a,1lpH-guaia-2-en-12,6a-olide

3a-Hydroperoxy-3-desoxoparishin A 259

3a-Hydroperoxy-8a-acetoxy-3-desoxo- 259 lip,13-dihydroparishin A

5a-Hydroperoxyasperilin 259

9B-Hydroxy-lB,1Oa-epoxypartheno1ide 216

3j3-Hydroxy-llB,13-dihydroacanthospermolide 150

llp-Hydroxy-11,13-dihydro-8-epi-confertin 261

9a-Hydroxy-ll, 13a-dihydrozaluzanin C 172

10|3-Hydroxy-lla-angeloyloxysIov-3- enolide 234

1lg-Hydroxy-13-chloro-l1,13-dihydroaromatin 134

la-Hydroxy-13-deacetyl-9,lOZ-dehydro- 130 1 ,10-dihydromarginatin

4a-Hydroxy-la,5aH-guaia-2,10(14), 259 11(13)-trien-12,6a-olide

3a-Hydroxy-2j3-senecioyloxy-7,8gH- 5 eudesm-4(15),11(13)-dien-8,12-olide

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

8a-Hydroxy-2-epi-vernomargolide-l,4- 130 cyclosemiacetal

9a-Hydroxy-3Z-seco-ratiferolide-5a-0- 67 [2 -methylbutyrate]

2-Hydroxy-4-acetoxyxanthan-l(5)- 67 en-6p,12-olide

3a-Hydroxy-4a,5a, 7a,1ip- 36 H-eudesman-12,Sp-o1ide

3a-Hydroxy-4a,5a-epoxyeudesm-ll- 92 en-12,Sp-olide

8p-4’-Hydroxy-5’-[4’’-hydroxytigloyloxy]- 192 tigloyloxy-3-dehydro-4p,15-dihydro

14-Hydroxy-5a-hydroperoxy-isoalantolactone 259

2a-Hydroxy-8p-2’ ,3’,5"-trihydroxy 190 angeloyloxycostunolide

2a-Hydroxy-8p-3' -hydroxy-2’,5’-epoxy 190 angeloyloxycostunolide

14-Hydroxy-Sp-[3-chloro-2-hydroxy 43 isobutyryloxy]-3-chlorodehydroleucodin

14-Hydroxy-8p-[4-hydroxytigloyloxy] 263 costunolide

2a-Hydroxy-Sp-isobutyryloxycostunolide 133

3-Hydroxy-8p-isobutyryloxydehydroleucodin 264

9a-Hydroxy-8p-methacryloyloxy-l4-0X0- 129 acanthospermo1ide

9a-Hydroxy-8p-methacryloyloxy-14- 129 oxo-acanthospermolide

14-Hydroxy-8p-methacryloyloxy-3- 43 chlorodehydroleucodin

3p-Hydroxy-8a-[2,3-diacetoxy-2- 127 methylbutyryloxy]-eremanthin

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

3p-Hydroxy-8a-[4-acetoxyangeloyloxy]- 127 eremanthin

3j3-Hydroxy-8a- [5-acetoxyangeloyloxy] - 127 eremanthin

3a-Hydroxy-8a-acetoxy-3-desoxo- 159 11^,13-dihydroparishin A

3p-Hydroxy-8a-epoxymethylacryloyloxy- 74 4(15),10(14),lip,13-dihydro-(laH),

4a-Hydroxy-8a-i sobutyroyloxy-4,5- 135 dihydro-5,6-dehydro-lO,13-bis-Q-methyl

4a-Hyd roxy-8a-tigloyloxy-lpH- 130 jalcaguaianolide-13-0-acetate

la-Hydroxy-9,102-dehydro-1, 10- 130 dihydromarginatin

15-Hydroxy-9a-methoxy-8/3-[2-methyl 127 butyryloxy]-14-oxo-acanthospermolide

9a-Hydroxy-seco-ratiferolide-5a-0-[2- 67 methylbutyrate]

9a-Hydroxy-seco-ratiferolide-5a-0- 67 isobutyrate

9a-Hydroxy-seco-rat if erolide-5a-0-angelate 67

5g-Hydroxyasper i1in 259

9-a-Hydroxyatripliciolide-8-0-isobutyrate 221

7a-Hydroxycostunolide 52

1Oa-Hydroxydams in 138

la-Hydroxydihydroridentin 65

20-Hydroxyelemajurinelloide 210

15-Hydroxyelemasteiractinolide 36

2a-Hydroxyeremophila-l(10),11(13)-dien- 36 12 ,8p-olide

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

3p-Hydroxyeremophila-l(10),ll(13)-dien- 36 12,8p-olide

3p-Hydroxyeremophila-9,11(13)-dien- 36 12 ,8p-olide

3a-Hydroxyeudesma-4,ll-dien-12.8g-olide 92

19-Hydroxyglaucolide E 119

7a-Hydroxygongrothamnolide 140

2a-Hyd roxyhanphy11in-3-O-acetate 259

20-HydroxyJurinelloide 210

8a-[4-Hydroxymethacryloyloxy]-10a- 119 hydroxyhirsutinolide

9^-Hydroxypartheno1ide 216

7a-Hydroxyreynosin 52

7a-Hydroxysantamarine 52

8a-[4-Hydroxytigloyloxy]-10a- 119 hyd roxyhirsutinolide

8p-[4-Hydroxytigloyloxy]-14-oxo- 263 4Z-acanthospermolide

8g-[4-Hydroxytigloyloxy]-14-oxo- 263 4Z-acanthospermolide

8p-[4-Hydroxytigloyloxy]-14-0X0- 263 acanthospermolide

8a-[4-Hydroxytigloyloxy]- 119 hirsutinolide-13-O-acetate

8g-5'-[4’’-Hydroxytigloyloxy]- 73 tigloyloxycostunolide

9^-Hydroxytournefortiolide 212

19-Hydroxyvernonatolide 140

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

6p-Hydroxyvernopappolide-8-0-methacrylate 140

9a-Hydroxyzaluzanin C 172

Idomain 75

8 -epi-Inuviscolide 261

1-epi-Inuviscolide 213

Iso-seco-tanapartholide 113

8/3-Isobutyryloxy-4-epiartecalin 264

8g-Isobutyryloxy-9g-hydroxy-10-acetoxy 203 methylgermacradien-6 ,7-olide

8oc-1 sobutyry loxyarmef o 1 i n 53

8a-Isobutyryloxyarmexifolin 53

8/3-Isobutyryloxyartecalin 264

8oc-1 sobutyryloxybalchanin 53

Isochiapine B 186

Isodehydroleucod in 226

Isodeoxyelephantopin 265

Isolasolide 234

Isosilerolide 110

8p-Isovaleryloxy-10a-hydroxy-l-oxo- 128 germacra-4E,11(13)-diene-12,6a-olide

8p-Isovaleryloxy-3(3, lOa-dihydroxy-1- 128 oxo-germacra-4E,ll(13)-diene-12,6a-olide

Istanbulin A 245

Istanbulin B 245

Istanbulin F 91

Ixerin U 232

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

Ixerin V 232

Ixerin W 232

Jamaicolide B 179

Jamaicolide C 179

Jamaicolide D 179

Jurinelloide 210

Lactucopicriside 172

Lactuside A 172

Lactuside B 172

Ladibranolide 80

Lantanifoline 96

Leucanthanolide 187

Ligustrin-8-yl-(2,5-dihydro-5-hydroxy- 36 3- furancarboxylate)

Ligustrin-8-yl-(4-hydroxytiglate) 36

Melampodin D 153

8a-Me tha cryloyloxy-1Oa-hyd roxy-1.13- 135 bis-O-methyl hirsutinolide

8a-Methacryloyloxy-lOa-hyd roxy-13- 135 0 -methyl hirsutinolide

8p-Methacryloyloxy-8-desacyloxy- 43 pycnolide 3-0-acetate

8a-Methacryloyloxyarmexifolin 53

8a-Methacryloyloxybalchanin 53

8g-Methacryloyloxyternifolin 181

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

2p-Methoxy-2-deethoxy-8-0-deacyl 26 phantomolin-8 -O-tiglate

2 p-Methoxy-2 -deethoxyphantomolin 26

lB-Methoxy-4p,15-epoxymiller-9Z-enoiide 129

2p-Methoxydesoxytagitinin B [sic] 264

12-Methoxydihydrodehydrocostuslactone 58

Methyl picrotoxate 221

8a- (2’ -Methyl )butyryloxy-10/3,1 la- 234 diacetoxyslov-3-enolide

8a-(2-Methylacryloyloxy)-compactifloride U 5

8p-Methylbutyryloxy-9B-hydroxy-10- 203 acetoxymethylgermacradien-6 ,7-olide

Micrantholide 32

Micrantholide-15-0-[2,3-dihydroxy 32 isobutyrate]

Micrantholide-15-0-[4-hydroxymethacrylate] 32

Micrantholide-15-O-isobutyrate 32

Micrantholide-15-O-methacrylate 32

Miller-1(10)Z,8E-dienolide 129

Miller-1(lO)Z-enolide 129

Mi 1ler-9E-enolide 129

Mi 1ler-9Z-enolide 129

Montafrusin B 199

5B-Myrtenyl-4a, 5-dihydroatripl idol ide- 221 8-0 -isovalerate

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

14-0-(4-Hydroxysenecioyl)isoverno 140 cistifolide-8-O-angelate

14-0-(4-Hydroxysenec i oy1)verno 140 cistifolide-8-O-angelate

14-0-(4-Hydroxysenecioyl)verno 140 cistifolide-8-0 -(2 ,3-epoxyisobutyrate

14-0-(4-Hydroxysenecioyl)verno 140 cistifolide-8-O-methacrylate

14-0-Acetyl-9-desacetoxyverno 140 cistifolide-8 -O-methacrylate

14-0-Acetylprevernocistifolide-8-0-angelate 140

14-0-Acetylvernocistifolide-8-0- 140 (2 ,3-epoxyisobutyrate)

14-0-AcetylvernociStifolide-8-O-angelate 140

14-0-Acetylvernocistifolide-8-0-methacrylate 140

14-0-Acetylvernocistifolide-8-O-tiglate 140

3-0-Methyltithonin 17

13-0-MethylvernoJalcanolide 8-0-acetate 135

13-0-Methylvernojalcanolide 8 -0-acetate 135

13-0-Methylvernojalcanolide 8-0-isobutyrate 135

14-0-Senecioylvernocistifolide-8-0- 140 methacrylate

14-0-Senecioylvernocistifolide-8-0-angelate 140

14-0-Senecioylvernocistifolide-8-0- 140 (2 ,3-epoxyisobutyrate)

Onopordopicrin-4a,5p-epoxide 215

Orthopappolide methacrylate 121

Orthopappolide senecioate 121

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

Orthopappolide tiglate 121

1-0xo-10pH-eremophila-7(ll)-en-8a, 12-olide 168

3-Oxo-1Oa-hydroxy-8p-i sobutyry1oxy 264 guaia-4, ll(13)-dien-12,6a-olide

2-Oxo-1Oa-perox i-8p-i sobutyry1oxygua i a- 264 3.ll(13)-dien-12,6a-olide

8-0xo-2a-9-dihydroxy-trans,trans- 45 germacra-1 (1 0 ),4-dien-trans-6,12-olide

4-ÜXO-3,4-seco-ambrosan-6,12-olide-3-oic 238

9-0xo-3Z-seco-ratiferolide-5a-0-[2- 67 methylbutyrate]

l-0xo-6j3, 7a, llpH, 14g-methylgermacra- 189 4(5)-ene-12,6-olide

1-0x0-60,7a,110H-germacra-4(5), 10(14)- 189 dien-12 ,6 -olide

3-0xo-eudesma-4,ll-dien-12,80-olide 92

3-Oxo-eremophila-9, 11(13)-dien-12, 80-olide 36

9-Oxo-isodehydroleucodin 226

9-Oxo-seco-rat iferolide-5a-0-[2- 67 methylbutyrate]

9-Oxo-seco-ratiferolide-Sa-O-angelate 67

9-Oxo-tournefortiolide 212

3-Oxoeremophila-1(10),11(13)-dien-12,80- 36

1-Oxoeudesm-7(11)-en-8 ,12-olide 246

Oxytoriloide 117

Parthoxetine 186

3a-Peroxy-50-hydroxyisoalantolactone 92

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

3a-Peroxy-5a-hydroxyisoalantolactone 92

4a-Peroxy-eudesma-2, ll-dien-12,8|3-olide 92

3a-Peroxyeudesma-4,11-dien-12,Sp-olide 92

3a-Peroxyisoalantolactone 92

Picriside A 173

Picriside B 173

Picriside C 173

Poskeanolide 119

Prenanthelide A 166

Prenantheside A 166

Prenantheside B 166

Prenantheside C 166

Pro-1,4-dimethylazulene 235

Pseudoelephantopide 8-0-methacrylate 135

Pseudoelephantopide 8-0-tiglate 130

Quadrangolide 112

Repdiolide triol 157

Repensolide 131

Rudbeckiolid 139

Rupicolin A-8-O-acetate 92

Rupicolin B-8-Ü-acetate 92

7|3-(1S, 5-Dimethyl-4-hexenyl)-3’ aa, 158 3'aa,5",6 ',7',8 ",8 'a,9',9'aa-octahydro- 5,5'g,8 'ap-trimethylspiro[bicyclo[2. 2. 2]oct- 5-en-2, 3’(2’H)-naphtho[2,3-b]furan]-2’-on

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

7p-(lS,5-DimethyI-4-hexenyl)-3’ aa, 4’, 4’aa,5’,6 ’,7’,8 ’,8 ’a,9’,9’aa-decahydro- 5, 8 ’ ag-dimethyl-5"-methylenspiro[bicyclo [2. 2. 2]oct-5-en-2,3'2’H)-naphtho[2, 3- b]furan]-2 ’-on

Saloniteno1ide 8-0-[1-acetoxyethy1aerylate] 258

Salonitenolide 8-0-acetyl sarracinate 258

Salonitenolide 8-0-angelate 258

Schkuhridin A 57

Schkuhridin B 57

Scorpiolid

Septuplinolide 178

Shonachalin D 231

Sonchuside A 165

Sonchuside B 165

Sonchuside D 165

Stilpnotomentolide 8-0-angelate 135

Subexpinnatin B

Subexpinnatin C

Tagitinin F-3-O-methyl ether

Tannunolide A

Tannunolide B

1, 2, 4, 15-Tetradehydro-4,5-dihydro-3- ososteiractinolide

8a-Tigloyloxy-2-epi-vernomargolide-l, 4- cyclosemiacetal

8a-Tigloyloxy-2-epi-vernomargolide-l,5- cyclosemiacetal

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Appendix B (continued) Compound name versus reference

8a-Tigloyloxy-2a,3^-dihydroxy- 4a-epoxy 21 dehydrocostuslactone

3a,4p-epoxide 8a-Tigloyloxy-4a,5a-epoxy 130 jalcaguaianolide

8a-Tigloyloxy-vernomargolide 130

8a-Tigloyloxyhirsutinolide-13-0-acetate 119

8(3-Tigloyloxypreeupartundin 33

Tomenphantopin A 99

Tomenphantopin B 99

Torilolide 117

Vernocinerolide-8-0-[4-hydroxymethacrylate] 119

Vernojalcanolide 8-0-acetate 135

Vernopappolide 140

Vernopappolide-8-O-methacrylate 140

Vernopappolide-8-O-tiglate 140

Vernopatensolide 8-0-angelate 135

Versicolactone B 265

Vicolide D 197

Youngiaside A 3

Youngiaside B 3

Youngiaside C 3

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Marta Vasquez was born on July 16, 1946 in Medellin, Colombia.

She attended Coiegio El Carmelo in Medellin, where she completed

education, through high school, in 1963. In 1964, she entered the

school of Architecture at the Universidad Nacional. In 1965, she

entered the school of Chemical Engineering at the Universidad de

Antioquia where she acquired a B. S. degree in Chemical Engineering in

1971. During her last year of College she held the position of

teaching assistant in physics, at the College of Basic Sciences of

the same University. She moved to Pereira in 1971 to fill a position

as professor in mathematics at the University of Pereira. In

1972-1973 she was a professor at the Basic Sciences College at

Universidad Nacional in Medellin. In 1974 she moved to Manizales

where she accepted a position as professor in organic chemistry at

the Universidad de Caldas. She went to Cali in 1979 and entered the

School of Chemistry at the Universidad del Valle where, in 1981, she

acquired a M. S. in Chemistry. In 1984, she came to Baton Rouge and

entered Graduate School at the Louisiana State University. She is a

member of the Phytochemical Society of North America. She is at the

present, a candidate for Ph.D. in the Department of Chemistry.

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. DOCTORAL EXAMINATION AND DISSERTATION REPORT

Candidate; Marta vasquez

Major Field: Chemistry (Analytical)

Title of Dissertation: Structure Elucidation of Secondary Metabolites from Rudbeckia Species by Spectroscopic Techniques and Review of Sesquiterpene Lactones.

Approved:

Major Professor and Chairman

EXAMINING COMMITTEE:

1 'c/d (AA''X£-/~'

Date of Examination:

July 17/ 1989

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.