Gyula Mózsik, András Dömötör, Tibor Past, Viktória Vas, Pál Perjési, Mónika Kuzma, Gyula Blazics, János Szolcsányi Capsaicinoids, From the Plant Cultivation to the Production of the Human Medical Drug

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CAPSAICINOIDS From the Plant Cultivation to the Production of the Human Medical Drug




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CAPSAICINOIDS From the Plant Cultivation to the Production of the Human Medical Drug By Gyula Mózsik, András Dömötör, Tibor Past, Viktória Vas, Pál Perjési, Mónika Kuzma, Gyula Blazics, János Szolcsányi






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The publication of this book was supported by the grant of the National Office for Research and Technology “Pázmány Péter Program” Ret-II, 08/2005 MEDIPOLISZ

ISBN 978 963 05 8694 8

© Gyula Mózsik, 2009

Published by Akadémiai Kiadó Member of Wolters Kluwer Group P.O. Box 245, H–1519 Budapest, Hungary

All rights reserved. No part of this book may be reproduced by any means or transmitted or translated into machine language without the written permission of the publisher.

Printed in Hungary



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Foreword . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 1. General introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 1.1. General introduction to the interactions of foods (or food components) and drugs to be used in the prevention and treatment of different diseases in patients. . . . . . . . . . . . . . . . . . . . . . . . . . 15 1.1.1. Foods and food components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 1.1.2. Drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 1.1.3. Levels of the drug–food interactions in healthy human beings and in patients with different diseases . . . . . . . . . . . . . . . . . . . . . 17 1.1.4. A short historic background of the interaction between the effects of capsaicin (capsaicinoids) and NSAIDs in animal experiments and healthy human subjects . . . . . . . . . . . . . . . . . . . . . . . . 18 1.1.5. Goals of the drug (drug-combination) production. . . . . . . . . . . . . . . . . . 20 2. Capsaicin (capsaicinoids) is (are) a famous family of the spices . . . . . . . . . . . . 21 2.1. Cultural background of spices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 2.2. History of spices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22 2.2.1. Terminology and modern history of Capsicum . . . . . . . . . . . . . . . . . . . . 23 2.2.2. History of Capsicum in Hungary. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 3. Botanical taxonomy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 4. Cultivation of Capsicum or paprika . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 4.1. Cultivation of Capsicums in India . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 4.2. Cultivation of Capsicums in other Asian countries . . . . . . . . . . . . . . . . . . . . . . 33 4.3. Cultivation of Capsicums in Africa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33 4.4. Cultivation of Capsicums in America . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33 4.5. Cultivation of Capsicums in Europe and Hungary . . . . . . . . . . . . . . . . . . . . . . 34 4.5.1. Cultivar types in Hungary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Sweet Capsicum cultivar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Paprika varieties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36




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5. General chemical structure and composition of Capsicums . . . . . . . . . . . . . . . 39 5.1. Macroscopic characteristics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 5.2. Microscopic characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 5.3. General chemical composition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 6. Chemical taxonomy of the functional parts of the Capsicums. . . . . . . . . . . . . . 43 6.1. Capsicum: Botanical aspects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 6.2. Capsicum: Chemical constituents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 6.2.1. Volatiles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 6.2.2. Coloring pigments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 Methods for determination of red and total carotenoids . . . . . . 53 6.2.3. Capsaicinoids. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 Chemistry of capsaicinoids. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 6.3. Capsicum: Quality control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 6.3.1. Capsicum fruits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 The European Pharmacopeia (Ph. Eur. 5.0) . . . . . . . . . . . . . . . . 61 6.3.2. Capsicum extracts – Oleoresin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64 Capsicum extracts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64 Capsicum Oleoresin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65 6.3.3. Quantitation of capsicum pigments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67 The Color Matching Method . . . . . . . . . . . . . . . . . . . . . . . . . . . 67 Spectrophotometric methods . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 The EOA Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 The American Spice Trade Association (ASTA) Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 The Hungarian Standard Method . . . . . . . . . . . . . . . . 68 6.3.4. Quantitation of pungent principles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 Official methods for organoleptic determination of pungency . . . . 69 The Scoville Method. . . . . . . . . . . . . . . . . . . . . . . . . . 69 The EOA Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70 The British Standard Method . . . . . . . . . . . . . . . . . . . 71 The International Standard Organization (ISO) Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 The ASTA Method . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 Quantitation of capsaicinoids . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 Early direct methods . . . . . . . . . . . . . . . . . . . . . . . . . . 72 Methods based on separation of capsaicinoids. . . . . . 72 Newer chromatographic micromethods – Thin Layer Chromatography . . . . . . . . . . . . . . . . . . . 73 Newer chromatographic micromethods – Gas Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . 73 Newer chromatographic micromethods – High Performance Liquid Chromatography . . . . . . . . 75 The ASTA method for determination of capsaicinoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77 The United States Pharmacopeia (USP) . . . . . . . . . . . 77 Correlation of pungency and capsaicinoid content . . . . . . . . . . 79 Stability of capsaicinoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80




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7. General physiology of retinoids and carotenoids. . . . . . . . . . . . . . . . . . . . . . . . . 81 7.1. Biochemistry of retinoids and carotenoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81 7.1.1. General physiology of retinoids and carotenoids . . . . . . . . . . . . . . . . . . 82 7.1.2. Retinoids and chemical carcinogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . 83 7.1.3. Effect of antioxidants on colorectal epithelial cell proliferation, polyp recurrence and carcinogenesis: Clinical trials in patients . . . . . . . 84 7.2. Prevalence and importance of the checked human GI diseases. . . . . . . . . . . . . 85 7.3. Gastric cytoprotection, as a special form of defensive mechanisms to gastrointestinal (GI) mucosal injury produced by retinoids . . . . . . . . . . . . . 90 7.4. New results in the biological actions of retinoids and carotenoids in animals, healthy human subjects and patients with different gastrointestinal disorders . . . 91 7.4.1. Gastrointestinal mucosal protective effects produced by retinoids in animal experiments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92 7.4.2. Effects of vitamin A on the gastric secretory responses and indomethacin-induced gastric microbleedings in healthy human subjects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92 7.4.3. Ulcer healing effect of vitamin A in patients with chronic gastric ulcer (multiclinical randomized, prospective study) . . . . . . . . . . . . . . . . 93 7.4.4. Changes in serum levels of retinoids in patients with different gastrointestinal inflammatory diseases . . . . . . . . . . . . . . . . . . . . . . . . . . 93 7.4.5. Changes in serum levels of retinoids in patients with different gastrointestinal cancers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Leiden mutation in patients with different gastrointestinal tumors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97 Correlation between the prevalence of Leiden mutation and decrease of serum levels of vitamin A and zeaxanthin in patients with different gastrointestinal tumors. . . . . . . . . . . . 97 8. Animal and human observations with capsaicinoids . . . . . . . . . . . . . . . . . . . . 100 8.1. Physiological and pharmacological research tool by capsaicin (Short overview) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100 8.1.1. The chemistry of capsaicinoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100 8.1.2. Source of capsaicin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101 8.1.3. Selective sensory effects of capsaicin . . . . . . . . . . . . . . . . . . . . . . . . . . 101 8.1.4. Mechanism of action of capsaicin on sensory receptors. . . . . . . . . . . . 102 8.1.5. Capsaicin actions in the gastrointestinal tract of animals . . . . . . . . . . . 103 8.1.6. Capsaicin-sensitive sensory nerves and gastric acid secretion . . . . . . . 105 8.1.7. Molecular-pharmacological studies . . . . . . . . . . . . . . . . . . . . . . . . . . . 107 8.1.8. Capsaicin actions in healthy human subjects and in patients with different gastrointestinal disorders . . . . . . . . . . . . . . . . . . . . . . . . 113 Results of the comparative molecular-pharmacological studies of capsaicin, atropine, omeprazole, famotidine, ranitidine and cimetidine on the gastric basal acid output (BAO) in human subjects. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115 8.1.9. Side effects of aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs) in the gastrointestinal tract of patients . . . . . . . . . . . . 118




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9. Toxicological studies with capsaicin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120 9.1. Animal observations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120 9.1.1. Acute toxicology studies of capsaicin in animal experiments . . . . . . . 120 9.1.2. Acute toxicity studies with pure trans-capsaicin derivates in dogs after intravenous administration . . . . . . . . . . . . . . . . . . . . . . . . 121 Acute effects on cardiovascular and respiratory parameters . . . 123 Plasma levels of capsaicin . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124 9.1.3. Results of subacute toxicology of capsaicin in dogs. . . . . . . . . . . . . . . 124 Two weeks treatment with trans-capsaicin . . . . . . . . . . . . . . . 124 Clinical chemistry and hematology . . . . . . . . . . . . . . . . . . . . . 124 Organ weights, macroscopic and microscopic observations . . . 125 Pharmacokinetic data after 14-days treatment with trans-capsaicin in dogs. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125 Absorption and metabolism of oral application of capsaicinoids in animal experiments. . . . . . . . . . . . . . . . . . 125 Summary and conclusions of the administration of different doses of trans-capsaicin in acute and subacute experiments in dogs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126 Chronic toxicity studies in animals . . . . . . . . . . . . . . . . . . . . . 128 9.1.4. Metabolism of capsaicin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128 The potential routes of metabolism of capsaicin . . . . . . . . . . . 129 Enzymatic oxidative metabolism of capsaicin . . . . 129 Non-oxidative metabolism of capsaicin . . . . . . . . . 130 Role of metabolic activation in capsaicin-induced toxicity . . . . 131 9.1.5. Effects of capsaicin on xenobiotic metabolism and chemically induced mutagenesis and carcinogenesis . . . . . . . . . . . . . . . . . . . . . . . 133 9.1.6. Hepatoprotection of capsaicin in rats . . . . . . . . . . . . . . . . . . . . . . . . . . 134 9.1.7. Genotoxicity studies with capsaicin or trans-capsaicin . . . . . . . . . . . . 135 Ames assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136 Mouse lymphoma cell mutation assay . . . . . . . . . . . . . . . . . . 137 Mouse in vivo micronucleus assay . . . . . . . . . . . . . . . . . . . . . 137 Chromosomal aberration in human peripheral blood lymphocytes (HPBL) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138 Brief summary of the main results of observations with capsaicin in animals. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140 9.2. Human observations with capsaicin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 9.2.1. Observations with capsaicin in healthy human subjects . . . . . . . . . . . . 141 Dose-response curves of capsaicin in the human stomach acute observation. . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 Changes in laboratory parameters and complaints of healthy human subjects during the study with capsaicin . . 142 9.2.2. Subchronic observations with capsaicin in healthy human subjects . . . . 142 Two weeks treatment with capsaicin . . . . . . . . . . . . . . . . . . . . 142 Biochemical measurements and complaints in healthy human subjects during two weeks capsaicin treatment . . . . . . 142 9.2.3. Human chronic observations with capsaicinoids . . . . . . . . . . . . . . . . . 143 9.2.4. Preventive effects of capsaicin against the selective and non-selective inhibitory actions produced by nonsteroidal anti-inflammatory drugs on COX-1 and COX-2 enzymes . . . . . . . . . . 143 8



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9.2.5. Summary of the observations with capsaicin alone or in combination with selective and non-selective inhibition of COX-1 and COX-2 enzymes by nonsteroidal anti-inflammatory compounds (drugs) in animal experiments and in human observations . . . . . . . . . . . . . . . . 145 10. Nature and characteristics of the innovative drug research. . . . . . . . . . . . . . 147 10.1. Characterization of the innovative drug research . . . . . . . . . . . . . . . . . . . . 147 11. Pharmaceutical industrial research and development . . . . . . . . . . . . . . . . . . 157 11.1. Product design and development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158 11.1.1. General goals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158 11.2. Chemical-pharmaceutical aspects of the product development . . . . . . . . . 158 11.2.1 Pharmaceutical form . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159 11.2.2 Composition and quality of starting materials. . . . . . . . . . . . . . . . . 159 Active Pharmaceutical Ingredient (API) of the product . . . . 160 Excipients’ quality of PH EUR requirements. . . . . . . . . . 161 Coating powder mixture quality . . . . . . . . . . . . . . . . . . . . 161 11.2.3. Packaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161 11.3. Formulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162 11.3.1. Formulation steps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162 11.3.2. FP Specification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162 11.3.3. Samples for clinical trial . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163 Preparation of samples for clinical trial . . . . . . . . . . . . . . 163 Exposing, stability and packaging of planned final product . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163 11.4. Plant batch manufacturing and process validation . . . . . . . . . . . . . . . . . . . 163 11.5. Summary of the Chemical-Pharmaceutical development . . . . . . . . . . . . . 164 12. Clinical pharmacological studies with capsaicinoids alone and with combination of capsaicinoids with nonsteroidal anti-inflammatory drugs. . . . 165 12.1. Main aims of clinical pharmacology and its relation to the evidence-based medicine. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165 12.2. Our special scientific problems in the human clinical pharmacology of capsaicinoids alone and capsaicinoids with together application of aspirin, diclofenac and Naproxen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167 12.3. Principal schedules for the human Phase I–II studies with capsaicinoids alone and together with aspririn, diclofenac and Naproxen . . . . . . . . . . . . 168 12.3.1. Preparation of protocols for the human clinical pharmacological studies (including Phase I to IV). . . . . . . . . . . . . . . . . . . . . . . . . . . 168 Medical points of the preparation of the study protocols. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168 12.3.2. Control of the protocols by the National or Regional Clinical Pharmacological and Ethical Commitees.. . . . . . . . . . . . . . . . . . . . 169 12.3.3. Pharmacokinetic and pharmacodynamic effects of capsaicinoids only. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169 Human Phase I clinical pharmacological study. . . . . . . . 169 Human clinical pharmacological Phase I study with capsaicinoids plus nonsteroidal anti-inflammatory drugs in healthy human subjects . . . . . . . . . . . . . . . . . . . 170




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Page 10 Human clinical pharmacological phase I study with capsaicinoids plus aspirin . . . . . 170 Human clinical pharmacological Phase I study with capsaicinoids plus diclofenac. . . 170 Human clinical pharmacological Phase I study with capsaicinoids plus Naproxen . . . 171 Human clinical pharmacological Phase II studies in patients . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171 Human clinical pharmacological Phase II study in patients with thromboembolic diseases (myocardial infarction, stroke, thromboembolic events). . . . . . . . . . . . . . . . . 171 Human clinical pharmacological Phase II studies in patients with different degenerative locomotor diseases . . . . . . . . . . 171 Acknowledgements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 173 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175 Appendix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207 1. European Commission. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207 1.1 Opinion of the scientific committee on food on capsaicin . . . . . . . . . . . . . . . . 207 1.2. European Herbal Practitioners Association . . . . . . . . . . . . . . . . . . . . . . . . . . 213 1.3. Commission Decision of 18 May 2005 amending Decision 1999/217/EC as regards the register of flavouring substances used in or on foodstuffs (notified under document number C(2005) 1437) (Text with EEA relevance) (2005/389/EC) . . . . . . . . . . . . . . . . . . . . . . . . . . . 215 2. Medical products for human use: Common Technical Document (CTD) . . . 219 2.1. Foreword . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219 2.2. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219 2.3. Terminology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220 2.4. Presentation of European Marketing Authorisation Applications . . . . . . . . . 221 2.5. Presentation of Applications in the Mutual Recognition Procedure or Decentralised Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221 2.6. Presentation of Follow-up Measures, Specific Obligations and PSURs . . . . 222 2.7. Reformatting of dossiers of already authorised products . . . . . . . . . . . . . . . . 222 2.8. Presentation of the application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223 2.9. Administrative, regional or national information is provided in different Modules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223 2.10. Preparing and Organizing the CTD . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 224 2.11. Pagination and Segregation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225 2.12. Information about national administrative requirements . . . . . . . . . . . . . . . 226 2.13. Special guidance for different kinds of applications . . . . . . . . . . . . . . . . . . 226 2.14. Special guidance on herbal medicinal products . . . . . . . . . . . . . . . . . . . . . . 229 2.15. Variation of an ASMF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 230 2.16. European Certificate of Suitability of monographs of the European Pharmacopoeia (CEP) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 231 2.17. European Community Guidelines on Quality, Safety and Efficacy . . . . . . . 231 About the authors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 239



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The potential role(s) of the scientific research works is (are) extremely well accepted in our days, when the Establishment of European Union celebrated its 50th years anniversary (2007). Consequently the countries of the European Union have chosen a new and common pathway for the economical, social and scientific development in Europe for the forthcoming time period (hopefully for all of the countries and for many centuries). Seven countries (France, West Germany, Italy, Spain, The Netherlands, Luxemburg, Belgium) established the European Economical Committee in Rome, Italy in 1957, when Europe demonstrated clearly the signs of survival over the Second World War to other parts of the World. Later on, this European Economical Committee has incorporated other countries including the countries liberated from the Soviet power. The final aim of these European political and economical changes is to establish an eminent, economically, politically and scientifically stable European Union. Hungary was accepted and entered the European Union by January 1, 2004. Thereafter many assets of our life have changed significantly (including economy, policy, social life, science). We have to try to find the most prominent lines still going on from the life in Hungary, which help us to reach the streams of the European Union. The pharmacological research and industry in Hungary were internationally well accepted by the different countries of Europe, North America (USA, Canada), Asia (Japan, China, India, and Pakistan). The essential and key role of so-called “innovative pharmacological research” has been especially accepted by the Hungarian Research Departments (including the Hungarian Academy of Sciences) and by the Hungarian Government. The different Regional University Science Centers (Debrecen, Budapest, Szeged and Pécs) have been established in 2004 by the Hungarian Government. Our (Pécs) University has received a grant for carrying out the innovative pharmacological research in the time period of 2005–2008 (National Office for Research and Technology “Pázmány Péter Program, RET-II, 08/2005”). The members (altogether 21) of the research are working in the University of Pécs, First Department of Medicine and Pharmaceutical Chemical Institute, Medical Faculty, University of Pécs, Hungary and PannonPharma Pharmaceutical Ltd. (Pécsvárad, Hungary). These researchers never worked together in a common project. So we found ourselves before an extremely new challenge in our common innovative pharmacologi11



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cal research, considering the very short time period (from 2005 to 2008) of our grant and our common success to produce new drug combinations of orally applicable capsaicin (capsaicinoids*) alone, capsaicinoids plus nonsteroidal anti-inflammatory compounds (aspirin, diclofenac, Naproxen) for patients suffering from myocardial infarction, pain, degenerative joint diseases, topically applied itraconazole (as one of the antifungal drugs) and etacrinic acid alone or in some combination to be used as an eyedrop preparation for patients suffering from glaucoma. The scale of this innovative pharmacological research is very wide and multidisciplinary. Furthermore, we had a very short time to realize the development of these scientific and industrial aims. We had several different scientific, industrial, marketing problems in the realization of our innovative pharmacological research. We met firstly with the problems of the use of capsaicin as chemical compound used as drug material. Capsaicin can be obtained from the plants, and this compound does not represent a uniform chemical entity (chemically capsaicin contains at least 5 to 6 chemical components (which are similar chemically and act in the same manner). This compound was never used as an orally applicable drug. Furthermore, we had very limited knowledge on the chemical compounds used in the cultivation of plants for extraction of capsaicin (capsaicinoids). We have learned a great deal of scientific matters from the lawyers, authorities, medical, industrial and patent work experts. The main point was focused, in these problems, to obtain the preparate of capsaicin (capsaicinoids) from the different plants. The environmental factors (application of different chemicals) played an essential role in the cultivation of capsaicin-containing plants. Briefly, the capsaicin (capsaicinoids) is (are) of plant origin (but not chemically produced) chemical compound (compounds). Consequently, the acceptance of a chemical compound of plant origin cannot be separated from the applied chemicals to produce larger quantities of the chemical compound (compounds), which may be environmentally harmful. Furthermore, we planned to produce capsaicin-containing drug combinations for oral application. During our innovative pharmacological research, we met a lot of different (medical, industrial, technological, botanical, agriculture, chemical, pharmacological, national and international juridical and patent) problems. All of us worked hard. Our aim was to give a short (but complete) summary of our common works in the years between 2005 and 2008. We hope very much that the results of our scientific endeavour can help the works of those participating in innovative research. Pécs, Hungary, September 2008

The Authors

* Important note: The term of capsaicin is used in the text, however, capsaicinoids of natural origin are used during different studies. Sometimes the term capsaicinoids is used to emphasize their plant origin in industrial research; suitable methods are used in the human clinical pharmacology (phase I–II).




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Generally, books and monographs are published to summarize the actually most considerable results of the relevant research topics. We have also published a series of different monographs, important books and congress proceedings up to now. Our studies with capsaicin (capsaicinoids) have been started from the 1980s in animal experiments and from 1997 in healthy human subjects and patients with different gastrointestinal (GI) disorders. The main point of the capsaicin research can be summarized by mentioning that capsaicin modifies significantly the function of the “capsaicin-sensitive afferent nerves” (the effects depending on the doses of applied capsaicin). The application of capsaicin is able to produce gastrointestinal mucosal defense. The gastrointestinal mucosal damage can be produced by different noxious agents, including nonsteroidal antiinflammatory drugs, NSAIDs in animal experiments and humans. The gastrointestinal mucosal defense induced by small doses of capsaicin (against Indomethacin, ethanol) has been proven in healthy human subjects, under conditions of Helsinki Declaration and respecting the rules of the Good Clinical Practice (GCP). The nonsteroidal antiinflammatory drugs (NSAIDs) are widely used in the treatment of patients with acute coronary syndrome (ACS), myocardial infarction, thrombophilia, stroke (aspirin) and with different degenerative diseases of the locomotor system [selective cyclooxygenase (COX) I inhibitors and non-selective COX-1 and COX-2 inhibitors]. These drugs produced different gastrointestinal diseases (mucosal damage, ulceration, bleedings, and gastropathia). The results of animal experiments and human observations have proved that capsaicin (given in small doses) is able to prevent the NSAIDs-induced GI complaints (mucosal damage, ulceration, bleedings, etc.). Different drugs (anticholinergic and antigastrinic drugs, histamine 2-receptor antagonists, proton-pump inhibitors, and scavengers) are widely used in the everyday medical practice to prevent the NSAIDs-induced GI side effects. The stimulation of capsaicin-sensitive nerves, by the application of capsaicin in small doses, represents a new pharmacological approach to prevent NSAIDs induced GI mucosal damage in healthy human subjects and in patients with different GI disorders. The Hungarian National Office for Research and Technology has offered us a grant for doing “innovative drug research in humans”. We submitted our application to produce a new drug (or drug combinations) to be applied in the patients treated with NSAIDs. 13



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The participants of this multidisciplinary group – receiving the grant (National Office for Research and Technology “Pázmány Péter Program RET-II, 08/2005”) – came from the Department of the Pharmaceutical Chemistry, First Department of Medicine, Department of Ophthalmology, Medical Faculty, Pécs University, Hungary and from the PannonPharma Pharmaceutical Ltd, Pécsvárad, Hungary (including 21 researchers altogether, among them internist, ophthalmologist, chemists, chemical engineers, biologist, bioenergineers, pharmacologist, horticultural engineer). We have met considerably different problems of medical, chemical, technological, industrial research, national and international laws, and marketing during our research work. We all together started with the “innovative drug research” and we had to learn many new things (results) from the different medical, chemical, pharmaceutical, industrial research during a very short time in 2005, in the pathways of mutually induced group education (including the research, legal, ethical, drug industrial fields; the necessity of the contact to the National Institute of Pharmacy (Országos Gyógyszerészeti Intézet, OGYI); to evaluate successfully the problems given by the National Pharmaceutical Institute in the field of our “innovative drug research”; marketing fields, etc. In our days, the role of “innovative drug research” has been widely accepted not only in Hungary but all over the World. Consequently we had a very short time to find the successful possibilities in the pathways of this research field. We had to learn the possibilities and basic requirements for the international registration of three patents of the produced new drug (or drug combinations) in the year of 2007: 1. Production of a new drug (or drug combinations): small doses of capsaicin (as orally given drug) applied alone or in combination with aspirin, diclofenac and Naproxen; 2. Production of a topically applicable antifungal drug in form of solution and in nail polish; 3. Production of a new drug combination of etachrinic acid with other chemical compounds to be applied in the patients with glaucoma. This book indicates and details the cultural history of spices (capsaicin) in the World; species, taxonomy, chemistry, medical research (in animal experiments and human observations), pharmacokinetic and pharmacodynamic determinations of drug (or drug combinations), preparation of suitable clinical pharmacological protocols, which are used in human studies (phase I–II); the actuality of the related and valid national (Hungarian) and international (European Union, United States, Japan, India, China) laws which give the principal bases of new patents and marketing. This book represents the very effective application of human power of the researchers to the enormous spiritual and physical challenges of our country, the European Union and other parts of the World in the field of introduction of a new drug (or of drug combinations). The research in this field is an extremely hard and interdisciplinary work. We believe that the experiments of our “innovative drug research” can help others, who are participating in these types of the innovative and multidisciplinary research. Pécs, Hungary, September 2008

The Authors

1. General introduction 1.1. General introduction to the interactions of foods (or food components) and drugs to be used in the prevention and treatment of different diseases in patients 1.1.1. Foods and food components The prevention and treatment of the different human diseases is the final form of our medical activities to keep the health conditions in the human beings. Nutrition and pharmacological treatment are also incorporated in this field. Nutrition alone is very important (including the consume of different macro- and micronutrients, trace elements, etc. in terms of their quantities and preparations) representing the basis of sustainance of our life, by supplying the necessary energetical background. The production of foods, establishment of the new foods, their biological and nutritional qualifications, cultivation of plants to be used for food production, industrial background cover a very wide range of natural sciences (physiology, pharmacology, nutrition, agriculture, food industry, medicine) - in terms of prevention and treatment - and of the different social sciences, marketing, etc. (Mozsik, Figler, 2005). The correct evaluation of these questions represents very urgent and important scientific aspects in our personal life, as well as in the World. The spices (chemical compounds of plant origin) have been used to modify the use of the different foods since 9500 years ago in the human culture. These spices are different chemically; they produce different changes in the process of nutritional habits, and on the other hand, are able to produce different physiological processes in healthy human beings and in patients with different diseases. Capsaicin (functionally capsaicinoids) is (are) one famous chemical compound among the spices. The historical and scientific history of capsaicin (capsaicinoids) is especially interesting: 1. capsaicin (capsaicinoids) does (do) not represent an energy supply for the man; 2. capsaicin (capsaicinoids) is (are) able to modify the carbohydrate metabolism (Domotor et al., 2006); 3. the spreading of capsaicin (capsaicinoids) has changed significantly over the World during the last 9500 years (Govindajaran, 1985, 1986a-f; Govindajaran et al., 1977, 1987; Govindajaran, Sathyanarayana 1991); 4. the cultivation of plants containing capsaicin changed significantly from time to time, especially in the last two centuries (Govindajaran, 1985, 1986a-f; Govindajaran et al., 1977, 1987; Govindajaran, Sathyanarayana 1991); 5. the discovery of a very wide scale of capsaicin-induced physiological regulatory mechanisms (Roosterman et al., 2006);

6. the discovery of the specific stimulatory effect of capsaicin on the afferent sensory nerves ("capsaicin sensitive afferent nerves") (Jancso-Gabor, 1959; Jancso et al., 1968, 1985; 1987; Jancso-Gabor, Szolcsanyi 1972; Szolcsanyi, 1977, 1982, 1983a,b, 1984a,b, 1985, 1990a,b, 1993, 1996, 2004; Buck et al., 1982; Monseerenusorn et al., 1982; Fitzgerald, 1983; Chahl et al., 1984; Hori, 1984; Russel, Burchiel, 1984; Holzer, Sametz, 1986; Buck, Burks, 1986; Maggi, Meli, 1988; Holzer, 1988, 1990, 1991a,b, 1992a,b, 1998; Maggi et al., 1989; Holzer, Lippe, 1988; Holzer et al., 1989; Maggi 1995; Jancso-Gabor et al., 1997); 7. The application of capsaicin produces different physiological (pharmacological) regulatory steps (Szolcsanyi, 1990a,b, 1993, 1996, 2004); 8. The application of capsaicin offered a new research tool to approach the different biological regulatory mechanisms in animals and human beings (Mozsik et al., 2007b) and in patients with different diseases (Domotor et al., 2005, 2006, 2007).

1.1.2. Drugs The research and production of different drugs are the keystones both in the drug industry and in the human medical treatment. The progress of medical treatments was based on two main research lines: 1. The evaluation of the results of the "classical and historical" medical practice dominantly started from Asia (from the Chinese medicine). The application of these medicines was based on the efficacy of the medical treatment carried out; 2. The planning of chemically, pharmacologically, biologically active compound(s) for production of the new drugs to be used in the treatment of patients with different diseases (that has been dominating in the last decades). Our attention is focused on the nonsteroidal anti-inflammatory drugs (NSAIDs). The most famous compound of this drug family was produced by the Pharmaceutical Company of Bayer (Germany) in the 1880s (Aspirin). This compound is very widely used in the everyday medical treatment (as analgetic, antipyretic agent, and platelet aggregation inhibitor), however, its most important indication is in the prevention of myocardial infarction, stroke, thromboembolic events and prevention of different colorectal malignancies. The main actions of NSAIDs are the following: 1. inhibitory action on the platelet aggregation; 2. production of gastric mucosal damage chemically (in the pathway of inhibition of dissociation of weak acid in the presence of a strong acid); 3. inhibition of the bicarbonate secretion produced by the pancreas; 4. inhibition of the prostaglandin (PG) synthesis and 5. modification of the blood flow in the gastrointestinal tract by the inhibition of the PG synthesis. The number of patients suffering from myocardial infarction or from different degenerative locomotor diseases is very large. The application of aspirin is generally and internationally accepted in the prevention of the myocardial reinfarction (given in doses of 100 mg/day or in large doses of 300 mg/day in so-called Aspirin 16

insensitive patients). The administration of aspirin means a long-lasting treatment in all patients with cardiological and thromboembolic events. Patients suffering from different degenerative diseases of the locomotor system are treated with one of the compounds of NSAIDs. The actions and pharmacokinetic behaviors of the NSAIDs can be modified by the application of different foods (fats, aperitive compounds, aldehyde and keto-sugars, dietary fibres, etc.).

1.1.3. Levels of the drug-food interactions in healthy human beings and in patients with different diseases There is a specific field of the human nutrition (including that of both healthy subjects and patients) considering that we use generally several macronutrients (proteins, carbohydrates, fats, etc.) together. There is no question that the absorption, metabolism, excretion and utilization of the macronutrients taken alone differ significantly from those when they are consumed together. Furthermore, the complementation of macronutrients with each other is a basic nutritional question (e.g. complementation of cereals with beans). Clinicians were taught that these problems (related to the application form of macronutrients) significantly differ in healthy subjects and in patients with different diseases. The different diseases are able to modify the digestion (all of the physiological processes which are able to prepare the foods in a suitable form for absorption), absorption and post-absorptive phases (transfer of the drugs from the small intestine to the target organ, excretion by the different pathways, utilization, including their anabolism and catabolism). The analysis of these medical questions requires a thorough medical knowledge. The detailed analysis of digestion, absorption and utilization of the different foods covers the main topics of the medical sciences (such as gastroenterology, nutrition, medical pathophysiology, organspecific topics of medicine as hepatology, nephrology, hematology, as well as different other subjects). Different drugs are used in the medical practice to modify the different physiological and pathological processes in healthy human subjects and in patients. Drug application to healthy subjects causes the transitory modification of the different functions in healthy human subjects (pain killers, coffee, laxative agents, many other drugs). Drug application in the patients with different diseases originally aims at decreasing the complaints of the patients. We have to know that these drugs modify the consumed foods (their absorption, digestion, utilization, transportation in the human body) (Table 1). A few examples will be given to explain these questions: 1. The prokinetic drugs decrease the transit time of the different foods in the GI tract of healthy humans, just like diarrhea (as disease). Consequently the time for the absorption of different foods is short; 2. Vitamin B is absorbed together with its intrinsic factor from the distal part of the small intestine. If the patients have no intrinsic factor, then the B absorption becomes insufficient, however, in patients with ileitis terminalis B absorption l 2

) 2

1 2


T a b l e I . T h e scientific a p p r o a c h of t h e m a i n q u e s t i o n s o f c l i n i c a l nutrition a n d dietetics a p p e a r s as f o l l o w s ( M o z s i k , Figler, 2 0 0 5 ) * 1.


F o o d a n d f o o d i n t e r a c t i o n s at t h e l e v e l s of 1.1.

pre-absorptive p h a s e (digestion),


absorptive phase,


post-absorptive p h a s e .

T h e d r u g a n d f o o d a c t i o n s w i l l m a n i f e s t at t h e l e v e l s of 2.1.

pre-absorptive l e v e l (digestion),


absorptive level,


post-absorptive l e v e l .

* Mozsik, Gy., Figler, M. (2005): Metabolic Ward in Human Clinical Nutrition and Dietetics. Research Signpost, Kerala (with permission)

becomes insufficient (partly due to the extremely decreased transit time) without any absence of vitamin B and its intrinsic factor; 3. Most of the macronutrients (carbohydrates, proteins) are absorbed by an active transport process from the small intestine. Strophantin g (and other digitalis compounds) specifically inhibits the active transport processes in the human gastrointestinal tract. In non-tropical sprue (gluten sensitive enteropathy) the patients have no brush border in the small intestine and consequently the mentioned food components will not be absorbed from the GI tract (without or with application of any kind of digitalis compounds); 4. Finally, the application of different drugs (nonsteroidal anti-inflammatory compounds) can modify the transport of food by the modification of the linkage of nutritional compounds and hormones to albumin, the excretion (e.g. diuretics) and the function of the target organ (positive and negative enzyme inductors). Moreover, in patients with hypalbuminemia (when the drugs are linked in smaller quantities to albumin) we can produce easily drug intoxication in the patients during different drug treatments. These examples from the everyday medical practice clearly indicate the extreme complexity of the correlation between nutrition and drug therapy in healthy human subjects and in patients with different diseases. 1 2

1.1.4. A short historic background of the interaction between the effects of capsaicin (capsaicinoids) and NSAIDs in animal experiments and healthy human subjects The mechanism of action produced by capsaicin (capsaicinoids) is very wide both in animal experiments and human beings (including the studies with isolated and cultivated cells, in vitro and in vivo studies, and whole living individuals) (Roosterman et al., 2006). 18

Our attention is focused on the mechanisms of action of NSAIDs and capsaicin both in animals and human beings. There is a vast literature dealing with the gastrointestinal mucosal damage produced by NSAIDs. The mechanisms of the NSAIDs-induced gastrointestinal mucosal damage can be detected at the levels of: 1. physico-chemical laws (Davenport, 1970, 1973; Mozsik et al., 1981); 2. prostaglandin synthesis (many authors worked in this field); 3. cellular energy systems (Mozsik et al., 1981; Mozsik, 2006); 4. oxygen free radicals (Mozsik, 2006). Szolcsanyi and Bartho (1981) were the first who proved that capsaicin prevents the gastric mucosal damage, when it is given in a small dose. In contrast, the gastric mucosal damage is enhanced by the application of higher doses of capsaicin. These results clearly demonstrated that the effect of capsaicin on the gastrointestinal tract depends on its doses applied in the experiments. After this original observation many researchers demonstrated the gastrointestinal preventive effect of capsaicin in animal experiments (Holzer, 1988, 1990, 1991a,b, 1992a,b, 1998; Holzer etal., 1989). A very systematic research work has been carried out by our team since 1980. Its main results and critical overview were summarized in a book (Mozsik et al., 1997a). The final conclusion of this book was that the application of capsaicin in the different animal experiments offers a new research tool for understanding the different mechanisms existing in the gastrointestinal tract. Because one of us (Gy.M.) is internist, nutritional expert and clinical pharmacologist, our attention turned toward the human physiological, pharmacological aspects of gastroenterology, the gastrointestinal tract and the mechanisms of actions of capsaicin related to other fields of medicine. The human clinical nutritional studies with capsaicin have been carried out from 1997, according to the Good Clinical Practice (GCP), respecting the Helsinski Declaration, and by the permission of the Regional Ethical Committee of Pecs University, Hungary. One of the main results of these studies was: Capsaicin (capsaicinoids) prevents (prevent) the Indomethacin (which is a nonselective cyclooxygenase enzyme inhibitor)-induced gastrointestinal side effect (microbleeding), however, it prevents the topically applied ethanol-induced decrease of gastric transmucosal potential difference (GTPD), and enhances the gastric transmucosal potential difference (GTPD) in healthy human subjects in association with gastric inhibitory actions on the gastric basal acid output (BAO) (Mozsik et al., 1999, 2004, 2005a, 2007b). These results offered a new possibility to plan and to create a new drug (or drug combinations), in which the gastrointestinal mucosal damaging NSAIDs are combined with the gastroprotective capsaicin in a suitable concentration and formulation.


1.1.5. Goals of the drug (drug-combination) production The NSAIDs are widely used in the everyday medical practice (prevention of thromboembolic episodes, myocardial reinfarction, stroke and colorectal cancers). These compounds act at the levels of enzymes of COX-1 and COX-2 by the modification of functions of these enzymes. The functions of COX-1 and COX-2 are involved in many therapeutic steps, new pathological events in the development of different diseases (myocardial infarct, damage in the gastrointestinal tract, development of different gastrointestinal malignancies, drugs-induced gastrointestinal injuries, etc.). The further (present) drug production research was based on our earlier obtained research works in the everyday clinical (cardiological, gastrointestinal, oncological) practice. The production of a new drug (or new drug combination) is a very hard and complicated work. This research is extremely well regulated internationally, covering the chemistry, animal experiments, stability studies, genotoxicology, metabolism, pharmacokinetic observations in animals and in human beings. The authors of this book participated in the establishments of clinical pharmacology (in the 1960s), clinical nutrition (in 1977) in the production of many drugs. These investigators work in the basic pharmaceutical research as well as in clinical nutrition, clinical pharmacology and medicine. This research represented a new challenge for all of us. Because this so-called innovative drug research does not represent a "typical research" pathway to us (independently all of us participated actively in the classical medical and chemical research), we decided that we try to give the necessary information of this "innovative drug research" for those, who want to participate in this process. In the course of this process we have gained knowledge considering national and international new scientific information, organization of the "innovative research team" to estimate our national and international possibilities including our limits, preparation of patents, preparation of the preclinical dossier, organization of human phase I—II trials, medical problems, marketing, etc.).


2. Capsaicin (capsaicinoids) is (are) a famous family of the spices

2 . 1 . Cultural background of spices Since the time man started gathering food, the addition of small amounts of some plant materials may have been used for the powerful impact they had on the appearance or eating quality. Initially, these additions were valued for masking the off-flavor of stored, decomposing foods; later on, in medieval Europe, they were imported as rural spices for their capacity to slow down deterioration during storage. Gradually, when definitive ethnic cuisines developed, the individual spices came to be valued in the modern sense for their real contribution, which is to flavor generally insipid meat and cereal foods and to make the food more acceptable and preferred. Spices have become indispensable in the modern kitchens of individual homes, institutions, and the food manufacturing industries. They provide individually to the dishes and distinguishing gourmet foods. In the countries of their origin and growth, the Hindustan, the Spice Island, and China, the highly systemized use of spices and other aromatic plants in foods, for increased acceptance and physiological effects of the extracts, has been known through ancient Vedic texts and the Ayurvedic text of Susrata and Caraca. For several centuries Before Christ (B.C.) spices, along with silk, gold, and precious stones, were principal articles of trade between the East and West. As early as 1600 B.C., the Chinese brought spices along the Chinese coast and overland along Tibet to Kashgar. The Arabs then took the spices west to Baghdad, Alexandria, and Persian Gulf ports. These were hazardous journeys, and great mystery surrounded the origin, collection and transport such that the values of the spices were very great. Later, the spice route shifted to sea routes (and included products besides those of Gulf ports) and then to the merchants in Alexandria and Baghdad. Knowledge of spices and their uses reached Egypt and spread to the Mediterranean region. Hippocrates, the father of modern medicine, and Theophrastus, the Greek philosopher and botanist, wrote treatises on medicinal plants that included many spices. During the first few centuries A.D., many cities (Alexandria, Constantinople, Basra and Venice) underwent violent changes as they became focal points of the spice trade. The values of trade in pepper and a few other spices were so great to the economy of the kingdoms in Europe that kings sent costly expeditions to spice-growing countries. In the late 15th century, Marco Polo of Venice, Vasco da Gama and Cabral of Portugal, and later Columbus and Cortes from Spain, undertook hazardous voyages 21

to establish sea routes to trading ports in the spice-producing countries of Asia. The trading became so intense that there were wars between Portugal, The Netherlands and England, while the local populations in Asia were forcibly enslaved and crippling taxes were levied. The entry of U.S.A. into the world of spices broke the monopoly of the European nations and the important trading centers shifted to Singapore and Calcutta in the East, and New York, London and Hamburg in West. The colonization of distant lands and the internationalization of the spice trade also had desirable changes, world trade in spices was increased and spices were introduced from one region to another. Black pepper, cardamom, ginger and turmeric from Asia were introduced into South America and Africa, and Capsicum (red pepper), vanilla and other spices were introduced from the New World of Central and South America to the Old World of Europe, Asia and Africa.

2.2. History of spices Among the spices, capsicums are the most colorful in appearance, and are important in terms of their history, antiquity and influence on many cuisines of the world. During his first expedition, Columbus (1492 to 1493) observed that the natives of the New World used a colorful red fruit calls aji or axi with most of their foods. This additive was found to be much stronger than the black pepper of Asia (Piper nigrum L.) in search of which he had undertaken this expedition. He took samples of this fruit back to Spain and named it "red pepper". De Cuneo, who accompanied Columbus on the second voyage to the New World in 1495, made the more definitive observations that "rose-like bushes have fruits as long as cinnamon, full of small grains, as biting as pepper; those Caribes and Indians eat that fruit like we eat apples". Chanca, the physician with the expedition, observed that the natives used this food additive both as a condiment and in medicine. Other travelers, who extensively traveled in Spanish America in the 16th and 17th centuries, described the popular use of Uchu, a colorful fiery pod of a plant, in the food of Peruvian Indians. Since early times, the internal use of a similar fruit as a cure for cramps and diarrhea by Mayan Indians of northern Guatemala has been recorded. The highly irritating smoke from the burning dried fruit is reported to have been used by the native Indians against Spanish invaders. For many years (centuries) prior to the Spanish occupation, the native Indians had been cultivating this piquant food additive. There was a considerable variation in color, shapes and sizes in the different regions. The great variety of capsicums used by the native tribes was thus known by different names, as mentioned earlier. Chilli or Chili, a name now commonly used in Asia and Africa, is said to have come from the Nahuatl dialect of Mexico and Central America. A wealth of original material thus became available and is still available in these areas for taxonomic and developmental work.


2.2.1. Terminology and modern history of Capsicum The term "chilli" is a rather confusing terminology: "chilli", "aji", "paprika", "chile", "chile" and Capsicum are all used frequently and interchangeably for "chilli pepper" under the genus of Capsicum, which belongs to a dicotyledonous group of flowering plants. A particular species of Capsicum is called "chili pepper" in parts of Mexico, southwestern United States and parts of Central America. To make matters still more confusing, a "sweet bell pepper" is often referred to as "Capsicum pepper", whereas to refer to the term "chilli", "hot pepper" is used. The term "bell pepper" is used to refer to a non-pungent, chunky, sweet chilli type, whereas "chilli pepper" generally refers to a pungent chilli variety. Red peppers have been familiar to all Spanish South Americans by the Arawakan name "aji" and by the Nahuatlan name "chilli" in Mexico and Central America. The genus Capsicum, which is commonly known as "red chile", "chilli pepper", "hot red pepper", "tabasco", "paprika" and "cayenne", belongs to the family Solanaceae (Night shade family) that includes tomato, potato, petunias and tobacco (Hawkes et al., 1979; Macrae, 1993). According to some references, the popular name "chili" or "chilli" originates from the hot pepper specially cultivated in the South American country, Chile. However, the name "chilli" seems to have nothing to do with the country name, on the contrary, it is believed to have originated from a district of Central America (De, 1992, 1993, 1994, 2000). One of the very first sources of life among the Inca, the "Commentaries Reales" by Garcilaso de la Vega, "El Inca", in 1609, mentions the common or even daily use of "chillies". According to de la Vega, there are three different kinds of chilli, two of which can be identified as "aji" and "rocoto", while the third is only insufficiently described. Even though the chilli is referred to by different names within the same country, and even in different states or provinces, the botanical name of chilli is the Latin name Capsicum. The word comes from a Greek based derivate of Latin "Kapto" meaning "to bite". The word "chili" is a variation of "cil" derivated from the Nahuatl (Aztec) dialect, which referred to plants now known as Capsicum, whereas "aji" is a variation of "axi" from the Arawak dialect of the Caribbean. This brings us up to the point of the correct way to spell "chile" (Domencini, 1983). The "e" ending in chile is the authentic Hispanic spelling of the word, whereas English linguists have changed the "e" to an "i". From the Nahuatl dialect of the Aztec language, the name "Chilrepin" has been derived. This was the name given to the earliest known word chile, the combination resulting in "Flea Chile", which is believed to allude to the sharp biting taste of the chilli pepper. Down the ages the original name has been slowly reformulated as "chile + reprintl" to "chilecping" to "chilrepin" to "chilepiquin", the latter two names being frequently interchangeable. The version used depends upon the source of information. However, Capsicum annuum var. aviculare is the modern scientific name of this earliest known variety (Macrae, 1993). However, a multilinguistic nation (like India) represents a unique case, where the same specimen Capsicum annuum L. is referred to by different names in different parts of the nation. In original Sanskrit the plant is known as "Mairichi phalam 23

and "Bruchi". However, in modern Indian languages it is known by different names, and even by more than one within the same language as given in Table 2. T a b l e 2. S u m m a r y of C a p s i c u m s in t h e l a n g u a g e s of different n a t i o n s * Language

Common name


Fructus C a p s i c i a c e r


Felfel, B i s b a s , F u l f u l h a l u


Mit'mita, Berbere

Assam i



C h e r v e n piper


Lanka, M o r i c h


N g a yut thee, Nil thee






S p a a n s e peper, C a y e n n e p e p e r


C a y e n n e pepper, R e d pepper, C h i l l i , Chili


Kibe paprika




P o i v r e r o u g e , P i m e n t e n r a g e , P i m e n t fort, P i m e n t - a i s e a u , P o i v r e d e C a y e n n e


R o t e r Pfeffer, C a y e n n e - P f e f f e r , Chili-Pfeffer, B e i f t b e e r e


Lai m a r c h a (red), Lila m a r c h a (green)


Pilpel adorn


Lai m i r c h (red), H a r i m i r c h (green)


C s i l i p a p r i k a , Igen eros a p r o , C a y e n n e bors, C a y e n n i b o r s , M a c s k a p o c s paprika, Aranybors, Ordogbors, Chilipaprika


Chilipipar, Cayennepipar


Lombok, Cabe, Cabai


P e p e r o n e , D i a v o l e t t o , P e p e r o n c i n o , P e p e di C a i e n n e , P e p e rosso p i c a n t e






M a k p h e t kunsi


Lada merah




Lai m i r c h y a (red), H i r v y a m i r c h y a (green)






P i m e n t a o , Piri-piri, P i m e n t a d e c a i e n a




Perets krasni


Marichiphala, Ujjvala


Rathu miris, Gasmiris


C h i l e , G u i n d i l l a , C a y e n a inglesa, P i m i e n t a d e C a y e n a , P i m i e n t a p i c a n t e , Ajf


Table 2. Cont'd Language

Common name


Pilipili h o h o




S i l i n g l a b y o , Sili






Pisi h u i , Prik k h e e , Prik


S i p e n m a r p o , Si p a n d m a r p o


A c i kirmizi biber





* After the paper of Basu, S. K., De, A. K. (2003): Capsicum: Historical and Botanical Perspectives. Taylor and Francis Ltd., London (with permission)

The chilli pepper is one of the very old domesticated plants of Middle America. In the Valle de Tehuacan (Puebla) which is one of the best documented examples of an early settlement in Mesoamerica, archeological evidence for the consumption of chilli peppers dates back to the seventh millennium B.C., long before the cultivation of maize and beans. The early findings of peppers - in coprolites and charred remains were probably harvested in the wild. However, domesticated chillies similar to their modern varieties in both size and shape can be found from the fifth millennium B.C. onwards. In pre-Columbian Mexico, chilli was one of the preferred tributes which dependent city-states had to deliver to their hegemonial powers. The paying of tribute also in the form of chilli peppers was later continued by the new Castilian rules. Capsicum has been known since the beginning of civilization in the Western Hemisphere. It has been a part of the human diet since about 7500 B.C. (MacNeish, 1964). It was the ancient ancestors of the native people, who took the wild chilli piquin and selected the values types known today. Heiser (1967) stated that between 5200 and 3400 B.C., the native Americans were growing chilli plants. This places chilli among the oldest cultivated crops of the Americas. Seeds found in early dwellings indicate that the natives were enjoying the peppers in 7000 B.C., along with potatoes in the Andes. In Mexico dry pepper fruits and seeds were received from 9500-year-old burials in Tamaulipas and Tehuacan. Domestication might have taken place 10,000 to 12,000 years ago. Christopher Columbus is believed to be the first European to discover chilli during one of his legendary travels to America around 1493. He was looking for an alternative source of black pepper which at that time was the favorite spice of Europe. What he discovered was a small fiery pod that for centuries provided seasoning for native Americans, the hot chilli pepper. It has to be emphasized that chilli or capsicum is not related to the Piper genus, which contains Piper nigrum L. of the family Piperaceae, the source of the black and white pepper. Within a century after its discovery hot chilli pepper attained a worldwide distribution. According to some 25

other sources, the American origin of Capsicum was first reported in 1494 by Chanca, a physician who accompanied Columbus in his second voyage to the West Indies (Macrae, 1993). Chilli peppers grow as a perennial shrub in suitable climatic conditions. The genus usually represents glabrous, perennial, woody subshrubs, some tending to be vines, rarely herbs, which are native to Central and South America, live for a decade or more in the tropical conditions of their natural habitat, but are mostly cultivated as annuals elsewhere. Chilli is native to the Western Hemisphere and probably evolved from an ancestor from the area of Bolivia and Peru. The first chillies consumed were probably collected from wild plants. Prehistoric Americans took the wild chilli "Piquin" and selected it for the various pod-types known today. However, domesticated chilli was apparently not grown prehistorically in New Mexico (Macrae, 1993). In fact, it is not known exactly when chillies were introduced into New Mexico. Chillies may have been used by the indigenous peoples as a medicine, a practice common among the Mayans. By the time the Spanish arrived in Mexico, Aztec plant breeders had already dozens of varieties. Undoubtedly, these chillies were the precursors to the large number of varieties found in Mexico today. Whether chillies were traded and used in New Mexican pueblos is still not clear (Macrae, 1993). Capsicum species have been thought to be of Central American origin, but one species has been reported to be introduced in Europe in the fifteenth century. By the middle of the seventeenth century, Capsicum was cultivated throughout Southern and Middle Europe as a spice and/or medicinal drug. One species was introduced into Japan and about five species were introduced into India, of which Capsicum annuum L. and C. frutescens L. were cultivated on a large scale (The Wealth of India, 1992). In commerce the description given applies to various African commercial varieties and these and the Japanese variety are sold in the United Kingdom as chillies, while the larger but less pungent Bombay and Natal fruits are sold as Capsicums. Very large Capsicum fruits that resemble tomatoes in texture and are practically nonpungent are widely grown in Southern Europe as vegetables (Evans, 1996). Records of the prehistoric Capsicum species around burial sites in Peru indicate that the original home of the chillies may be tropical South America. These seem to have been diffused from there to Mexico, or an independent origin in the latter country, where a great diversity of the genus is found. While Capsicum annuum has not been recorded in the wild state and Capsicum frutescens doubtfully so, they have now naturalized in the tropics of many countries and are easily disseminated by birds (The Wealth of India, 1992). The plant was introduced into Spain by Columbus, from where it spread widely. Subsequently, the prolonged viability, easy germination, and easy transportation assisted its spread all across the globe. The original distributions of this species appear to have been extending from the South of Mexico into Columbia (The Wealth of India, 1992). "Ginnie Pepper" was well known in England in 1597 and was grown by Gerarde (Evans, 1996). The Portuguese introduced chilli into India. Chilli is used as a condiment in large quantities in India, Africa and tropical America, where the fruit develops greater pungency than in colder regions. It has now, however, become a popular condiment 26

all over the World. The long, thin fruits constitute the source of dry chillies used for commerce. The wide popularity of chilli and its extensive cultivation are due to its being a short duration crop and its ease of cultivation under a wide range of climatic and adaptive conditions, particularly in comparison with black pepper {Pepper nigrum L.). The cultivation and utility of both Capsicum spp. are similar, except for local peculiarities (The Wealth of India, 1992). Capsicums are mentioned in a classic text of the Tibetan medical tradition, the "Blue Beryll": "Capsicum (tsi-tra-ka) increases digestive warmth of the stomach, and is the supreme medication for the alleviation of edema, hemorrhoids, animalcules, leprosy and wind". Another passage tells us "bad-kan-nad-sel tsje-'phel tsi-tra-ka mar sbrang sbyar - to alleviate diseases of phlegm and prolong the lifespan: [use] Capsicum mixed with butter and honey". Interestingly, the spice translated as cayenne pepper has another name, Yer-ma, and is generally used to treat wind disorders. In pre-Hispanic medicine, chilli peppers were used to treat a host of conditions, often in combination with other plant and mineral substances. For example, chilli was used to treat diseases of the gastrointestinal tract (infections, diarrhoea), in addition to tooth pain, cough and lack of appetite. It was popular as an aphrodisiac, as well. For that reason, the Spanish Padre Acosta, traveling through New Spain in the sixteenth century warned that high consumption of the chilli peppers would be detrimental to the "soul's health" because it "promoted sensuality" (Acosta, 1985). Chilli peppers had other uses as draconic punishment in children's education, and even in warfare - the enemy was driven out of his fortification by the employing the acrid smoke of smoldering chillies. This tactic was employed in pre-Columbian times, but also during the Mexican revolution at the beginning of the twentieth century.

2.2.2. History of Capsicum in Hungary Spain cultivated the pimientos, valued for the brilliant red color, characteristic aroma, and mild pungency. These are more and more being produced as an industry with close state control, integrating the steps of cultivation, processing, quality control and marketing. In Europe, Hungary and the Balkan countries have been producing the carefully selected cultivars of paprika since the 18th century and possibly still earlier. Hungary and the Balkan countries have been the traditional producers of highly valued burgundy red, elongated conical, fleshy smooth erect fruit of paprika for its color and flower. The main areas of cultivation in Hungary are Szeged and Kalocsa alongside the Danube (Govindarajan, 1985). The first written records of the appearance of paprika in Hungary are from the middle of the sixteenth century. Paprika was grown as a rarity in the garden of Margit Szechy, the step-mother of the distinguished general Miklos Zrinyi. It can be assumed from the information of the next two to three centuries that Capsicum was cultivated only to be used as a condiment. The cultivation of the sweet Capsicum known today began at the end of nineteenth century. Bulgarians were the first growers who cultivated Capsicum in the southern part of the country (Szentes and surroundings) (Somogyi et al., 2003). 27

At the beginning of the twentieth century, production of sweet Capsicum spread to other parts of Hungary. It is interesting to note that the sweet Capsicum growing regions are not the same as the traditional paprika producing areas. Important sweet Capsicum areas were in the middle of the country (Boldog, Nagykoros, Cegled), and in the south-southeastern part (Gyula, Baja, Bogyiszlo). But today this typical vegetable is cultivated all over the country. The production of condiment paprika in Hungary is highly labor-intensive. It became a significant crop at the end of the nineteenth century, although reports of its cultivation exist from the sixteenth century in Szeged, where the crop was mentioned as one of the crops grown. It was introduced from Turkey by monks, who excelled in healing, and was used as an effective medicine against malaria. Looking at the history of Hungarian paprika production we can distinguish several classical periods: - Until the middle of the nineteenth century feudalistic family self-sufficiency and the beginning of the production for the market; - Until the beginning of the First World War paprika production is market-oriented based on free competition; - From the end of the First World War until the end of Second World War a stateregulated production order was characteristic; - Paprika milling was under state monopoly for the period of 1940-1990; - In the last decade of the twentieth century again free market: production and processing are based on competition. Spice made of paprika is known as "Hungaricum" worldwide, and is an essential element of the Hungarian cuisine. Until the turn of the nineteenth century it was known in public life mainly as medicine. Paprika (Capsicum annuum L.) originated from South America and came to Europe - probably first to Spain in 1493, after the discovery of the American continent (Pickersgill, 1986, 1989). From there it came to Hungary across the Balkan through Turkish growers. The first paprika plants were planted at the end of the 1500s. At first paprika was considered as an ornamental plant and was grown for culinary usage at the beginning of the seventeenth century. The plant later enjoyed tremendous popularity around the time of Napoleon (Somos, 1981). The book containing its first detailed description was written by Csapo (1775). According to this book it is grown in vegetable gardens and the long red fruit is dried and crushed to a powder. Veszelszky (1798) mentioned around that time that the farmers of Fot, Palota and Dunakeszi grew paprika. The first cultivation trials were conducted at the botanical garden of the University of Pest in 1788. Since that time different Capsicum varieties were found in the "Index seminum" of the botanical garden (Augustin, 1907). In letters that Count Hoffmansegg sent his wife about his journey in Hungary, he mentioned "here I really liked a Hungarian dish, meat with paprika. I must be very healthy" (Balint, 1962). Augustin, a German traveler did not talk so nicely about the Hungarian paprika in his book, "Die Ungarn wie sie sind" (1831). He called paprika "Diablische Paprika Briihe". He wrote that for people who are not used to it, the effect on the palate is like embers or even worse (see for details Somogyi et al., 2003). 28

3. Botanical taxonomy

Capsicum is a genus of the family Solanaceae and is closely related to another genus, Solanum, which covers many economically important plants such as the potato {Solarium tuberosum L.) eggplant {S. melongene L.) tomato (Lycopersieon esculentum Mill.), and tobacco (Nicotiana tobacum L.). Some 20 to 30 species of Capsicum are reported to have their origin in the New World covering the tropical area of Northern South America, Central America, Mexico, and the islands in that region. Many of these occur in the wild as natural growth in areas of undisturbed vegetation, though their fruits were collected and distributed throughout the marketplace. Exploration and plant collection throughout the New World had given us a general but false impression of specification in the genus. Humans selected several taxa and in moving them toward domestication selected the same morphological shapes, size and colors in at least three distinct species. The early explorations in Latin America were designed to sample the flora of a particular region. Thus, any collection of Capsicum was a matter of chance and usually yielded a very limited sample of pepper from that area. Only with the advent of collecting trips designed to investigate a particular taxon did the range of variation with species begin to be understood. One needs only to borrow specimens from the international network of herbaria to appreciate what a limited sample exists for most taxa, particularly for collections made prior to 1950. The domesticated Capsicum pubescent, e.g., that is wide-spread in the mid-elevation Andes from Columbia to Bolivia, is barely represented in the herbaticum collection of the world. The taxonomy of Capsicum represents a very complex approach to different species. Many classifications were published earlier (see Govindarajan, 1985; Basu, De, 2003). In 1957 Smith and Heiser recognized five species only, which gives the "Big Five" species of Capsicum. Further recent works by Eshbaugh, Pickersgill and Hunziker identified 22 other wild species (Macrae, 1993). The wild gene pool, tightly linked to the domesticated, is designated Capsicum baccatum var. baccatum and is most common in Bolivia, Brazil, Chile and Argentina. Its flowers are solitary at each node. The pedicels are erected or declining at anthesis. Corollas are white or greenish-white, with diffuse yellow spots at the base of corolla lobes or either side of middle-vein (the flower is white with yellowish spots, anthers are white but turn brownish-yellow with age): the corolla lobes usually are 29

slightly revolving. The calyx of mature fruit is without annular constriction at its junction with the pedicel (though sometimes irregularly wrinkled), veins prolonged into prominent teeth. The fruit flesh is firm. Seeds are straw-colored. Chromosome number is 2n=24, with one pair of acrocentric chromosomes (Escabeche, Peru). The following five major species are morphologically definable: Capsicum annuum var. annuum L., Capsicum frutescences L., Capsicum chinense Jacq, Capsicum baccatum var. pendulum wild, Capsicum pubescent Riuz and Pav (Table 3) T a b l e 3 . T h e m o r p h o l o g i c a l i d e n t i f i c a t i o n of t h e f i v e m a j o r s p e c i e s * Species

Flower color

Number flw/node

Seed color

Calyx constriction



















white with









y e l l o w spot C.



* After the paper Basu S.K., De, A.K. (2003): Capsicum: Historical and Botanical Perspectives Taylor and Francis Ltd., London (with permission)

The haploid chromosomal count of the cultivated and wild species is 12. There are wide variations among and within the species, whether wild or cultivated, as no karyotype is characteristic for any single species and certain characteristics are observed among the majority of the members. Natural polyploidy is reported in the case of Capsicum, although a spontaneous tetraploidy has also been reported in an intravarietal cross. Induced polyploidy with colchicine has also been reported where the induced polyploid exhibits a high vitamin C content profile. Diploids showing mitotic abnormalities and irregulaties have also been reported (The Wealth of India, 1992). Generally, there appears to be a well-developed sterility barrier between cultivated species. It is impossible to cross Capsicum pubescens with other species. Several crosses between Capsicum annuum, Capsicum frutescens and Capsicum baccatum var. pendulum have produced a few Fl hybrids, but are mostly highly sterile. In the case of favorable crosses, the success or failure depends on the direction of the cross. Reciprocal differences were observed in the case of fertility. The results of hybridization also differed according to the parental cultivars. Viable seeds have easily been produced from Capsicum annuum X Capsicum chinense and Capsicum frutescens X Capsicum pendulum. The crosses Capsicum annuum X Capsicum frutescens and Capsicum frutescens X Capsicum chinense have yielded a few viable F l , F2 and bud cross seeds (The Wealth of India, 1992) Pronounced differences have been found in the capsaicinoid composition of the individual species, and the chemotaxonomic basis for identification is in line with the earlier classification based on floral morphology and can therefore be used as an 30

additional method. The key for the identification of the individual Capsicum species is given based on the limits of the total and the three individual capsaicinoids and their properties in Table 4. T a b l e 4 . A c h e m o t a x o n o m i c k e y t o t h e i d e n t i f i c a t i o n of c u l t i v a t e d C a p s i c u m s * N D H C fraction o v e r 9 . 5 % 1.1 C fraction o v e r 5 6 %



1.2 C fraction u n d e r 5 6 %







2.1 C f r a c t i o n , 4 2 t o 5 7 %





2.2 C f r a c t i o n u n d e r 4 2 %














2 N D H C C Fraction 9 . 5 %

2.3 C f r a c t i o n , 5 7 t o 7 3 % D H C fraction 2 6 t o 3 4 % Total c a p s a i c i n o i d s , u n d e r 0 . 3 5 % 2.4 C fraction o v e r 6 3 % D H C Fraction u n d e r 3 2 % Total c a p s a i c i n o i d s , o v e r 0 . 3 5 %


* After Jurenitsch J . , Kubelka, W., Jentzsch, K. (1979c): Identification of Cultivated Taxa of Capsicum Taxonomy, Anatomy and Composition of Pungent Principles. Plant. Med. 35: 174-180 (with permission) Abbreviations: C-Capsaicin; NDHC-Nordihydrocapsaicin; DHC-Dihydrocapsaicin

Thus, on the basis of the above studies, one may come to the conclusion that the genus Capsicum represents a very wide and divergent taxonomic group consisting of both wild and cultivated species. Some workers consider Capsicum to consist of three principal species, C. annuum, C. frutescens and C. chinense; meanwhile others have divided the genus into a divergent spectrum of species. Probably, molecular biology and molecular genetics, phytochemistry and cell biology can contribute to a deeper recognition of the taxonomy status of the genus Capsicum.


4. Cultivation of Capsicum or paprika

Capsicums, indigenous to South and Central America, Mexico and West India, continue to be cultivated there and have been introduced and widely cultivated through temperate, subtropical Europe, the southern of United States of America, tropical Africa, India, East Asia and China. The cultivation - through the last 400 years in the different soils, cultivation conditions, natural hybridization and selections - has given us a variety of Capsicums. Valued solely for their color and secondarily for aroma and mild pungency are the Spanish and Hungarian paprikas. For color, medium to high pungency and aroma are the chillies grown in India, Southeast Asia, China and America, and essentially for pungency the chillies in tropical Africa. Typical practices of the cultivation of Capsicums are written briefly below.

4 . 1 . Cultivation of Capsicums in India Chillies have been cultivated in India for over 200 years and have spread to most of countries, from sea level to 1600 m above sea level with annual rainfall of 600 to 1250 mm. Throughout India, 14 states cultivate more than 10,000 ha each, and 3 of them over 150,000 ha. The major chilli-producing states are Andhra Pradesh, Maharasthra, Orissa, Karnataka and Tamil Nadu. Chilli is a warm season crop, but low humidity with high temperature results in the shedding of buds, flowers and young fruits. Very low temperatures also result in poor growth. Chilli is started in three seasons, in different parts of the country. The main crop on the southern plains in India - which produces 70% of the crop (total of nearly 500,000 tons annually - is started in May to June, transplanted in 5 to 6 weeks, and harvested from October. In the Punjab area the cultivation of crop starts at the end of the cold season (in March to April), to avoid damage from frost, and harvested in August. In the Gangetic plains the crops are started in September and harvested in February. Chilli is cultivated in many areas in the north as a rain-fed crop, but in the south it is increasingly cultivated as an irrigated crop, especially in Andhra Pradesh and Tamil Nadu. The crop is grown in many types of soils but well drained loamy soils rich in organic matter are considered the most suitable. Clayey loams which can retain moisture are those in which chilli is cultivated as a rain-fed crop. 32

A variety of chillies, combining different sizes and degrees of pungency, are grown in India, each area having their characteristic cultivar of Capsicum annuum var. annuum. Generally, the medium to long and medium to high pungency red varieties are grown in the plains and the long, deep red, thick-fleshed, low pungency types are grown at higher elevations. Some amounts of vegetable capsicums and red short blunt conical types are also grown. Minor amounts of the highly pungent, small conical variety of Capsicum frutescens are grown or collected from semi-wild growth. A typical cultivation in Tamil Nadu, India is as follows; seedlings are started and raised in carefully prepared beds with natural shade and near a water source.

4.2. Cultivation of Capsicums in other Asian countries In the Indian subcontinent, other notable producers are Pakistan and Bangladesh, where the cultivation practices are similar to that in India, in amounts of internal consumption, with only a small percentage of export of a short conical variety ("dandicut cherry"). The other consumers of a number of Capsicum annuum, with moderate to high pungency and good color, are Thailand, Indonesia and Malaysia. In Southern Asia, South Korea is a substantial producer of highly colored and rather high pungency chilli varieties, but with a very high consumption, it is not an exporter. Since the 1950s, China has emerged as a major producer. A Federal Agricultural Organization (FAO) estimate of the annual production of fresh chillies is put at 1 million ton. China has become a top exporter, equalling, and even exceeding India.

4.3. Cultivation of Capsicums in Africa In the African continent, the crop was introduced early by the European conquests, but largely remained as semi-wild plants and exists as an organized cultivation in small holdings in only the North and West African countries. Both medium-sized mild and high pungency small chillies have been produced. The mild form is preferred for local consumption, the export being dominated by the crop collected from the wild growth in East Africa, the cultivated Zanzibar type from Sierra Leone, and the mild to medium type from Kenya and Nigeria. Ethiopia is notable for the largescale cultivation of a variety of Capsicums, the mild to medium pungency red chillies and more recently, paprika for oleoresin production.

4.4. Cultivation of Capsicums in America In the Americas, the source of Capsicum, the Central and South American countries continue to produce a great variety of Capsicums by collection of wild growths and by rudimentary cultivation of specific local varieties mostly for internal consumption. The wild Capsicums are reportedly used fresh, and medium pungency types are dried product. The highly pungent small varieties from all the five species cultivated are used essentially for therapeutic use. Mexico alone in this group appears to pro33

duce mild and high pungency chillies in amounts to have a substantial surplus for export to the United States. It has been difficult to obtain information on organized cultivation from these countries. In the United States, Capsicums have been cultivated since the 1920s in California, South Carolina, Louisiana and New Mexico. Considerable developmental work on the selection of suitable cultivars adapted to this region optimal cultivation practices. Different cultivars which combine the qualities of color and pungency in the required proportions are grown and account for nearly half of the total requirement of chillies and paprika, both of which had been earlier wholly imported. Moderately pungent ("Louisiana sport", "Carolinas" or "Anaheim" in California), the high pungency ("Bahanian" in South Carolina and "Tabasco" in Louisiana), the green bell Capsicum ("California Wonder" and "Florida Giant" for the fresh market in Florida and New Jersey), paprika ("Ruby King" in California) and pimientos ("Perfection") in Georgia exist as flourishing crop, raised through modern cultivation and processing methods. Large-scale cultivation and processing practiced in the United States are based on the basic scientific information collected on the growth of capsicum plant, and the formation of functionally important components for optimized economic production. Considered as a warm season crop, Capsicums are cultivated in spring, summer and autumn in the upper south of the United States and even in the winter in the extreme south. Boswell recorded that the optimum temperature for growth is around 24°C, and night temperatures lower than 15°C or day temperature higher than 32°C are detrimental to growth and fruit set. Additionally, low humidity causes abscission of flowers, buds and small fruits. At least 3 months of warm weather are necessary for the quick maturing sweet vegetable Capsicums and 4 to 5 months for most other cultivars. The small-fruited forms, those of Capsicum frutescent, are much more tolerant of higher temperatures and also are late maturing, requiring a longer warm season. The Capsicums can be grown on well-drained soils, but a fertile loamy soil rich in lime is considered most suitable. They are generally grown in rain-fed areas having 600 to 1250 mm of rain annually and redden better under controlled irrigation. Heavy rain and water logging lead to poor fruit set or fruit rot.

4.5. Cultivation of Capsicums in Europe and Hungary In Europe, since the 18th century, Hungary and the Balkan countries have been producing the carefully selected cultivars of paprika and possibly earlier still, Spain has cultivated the pimientos, valued for their brilliant red color, characteristic aroma, and mild pungency. These are more and more being produced as an industry with close control, integrating the steps of cultivation, processing, quality control and marketing. Sweet Capsicum can be grown in any region of Hungary with the exception of the western border of the country where the precipitation is higher and the temperature is lower than average. Only 8 to 10% of the country's soil and climate conditions are suitable for growing sweet Capsicum. Nevertheless, as traditional growing regions evolved, immigrant Bulgarian market gardeners settled at the southern part 34

of the country and started vegetable production. However, in a significant part of the region we find higher, sandy loam. These soils have inadequate water management with less humus, the cultivation is easier and the soil warms up faster, at the same time the quality of the condiment paprika is lower than that obtained from the heavier soils. These lighter soils are suitable for intensive sweet Capsicum production under controlled conditions. In the region of Kalocsa the soil is mostly heavy, with relatively low humus content on the former flood plains of the Danube River.

4.5.1. Cultivar types in Hungary Sweet Capsicum cultivar The sweet Capsicum cultivars in Hungary have changed a lot during the last centuries, especially during the last 50 to 60 years. In the previous centuries the seed was handed down from father to son and the growers passed the seed on to each other. The seeds of the best and earliest fruits were always kept for further sowing. It can be assumed that on many occasions spontaneous pollination or mutation created new types or varieties. Breeding techniques applying genetics go back only to the last 50 to 60 years in Hungary. The first sweet cultivar which received State registration was "Cecei sweet 3" bred by Angeli (see Somogyi et al., 2003). He selected this from a pungent white type and created a non-pungent cultivar. With his work, an active and efficient Capsicum improvement started and consequently the Hungarian sweet Capsicum types and an assortment of cultivars have been continuously increased and spread all over the World. The production of sweet Capsicum hybrid seed cultivars was initiated in Hungary in the middle of the 1960s (Moor, 1969). Cultivars were made by the breeding work of the state-owned Vegetable Crop Research Institute and its processors, and the Horticulture Departments of Universities until the middle of 1980s. Since then, several private breeders' cultivars received Plant Variety Protection. All the cultivars grown belong to the species of Capsicum annuum L. The sweet Capsicum cultivars (varieties) are categorized into different groups by the National Institute for Agricultural Quality Control (Table 5). Approximately 100 cultivars were registered in the National List of Varieties in 2000. One of the most important trains of sweet Capsicum cultivation is the sensitivity to the lack of light (Zatyko, 1979). This characteristic is important for determining whether the cultivar can be force-grown or not. This means establishing whether the cultivar may be grown only in the field or whether its production is economical under green-house conditions out-of-season: 1. Ciklon Fl - indeterminate, white fruit ripening to red, sweet, conical, upright fruit, for all production systems. Fruits are 12 to 15 cm long and 5 to 6 cm in diameter. Yield is 8 to 15 kg/m depending on the production technology. It contains Tm2 resistance. Under local night conditions, sowing is done at the end of September and the first 2 cm long fruits appear after a vegetation period of 125 to 130 days. This can decrease by 50% under more intense light conditions; 2


T a b l e 5. C a t e g o r i e s of t h e H u n g a r i a n s w e e t C a p s i c u m c a t e g o r i z e d b y t h e H u n g a r i a n N a t i o n a l Institute for A g r i c u l t u r a l Q u a l i t y C o n t r o l * W h i t e fruit, i n d e t e r m i n a t e

p u n g e n t or w i t h o u t p u n g e n c y

W h i t e fruit, d e t e r m i n a t e

without pungency

Pale g r e e n fruit, i n d e t e r m i n a t e

without pungency

Hornshaped, indeterminate

pungent, without pungency

P o i n t e d , hot, i n d e t e r m i n a t e

without pungency

P o i n t e d , hot, i n d e t e r m i n a t e

without pungency


without pungency

California W o n d e r type

without pungency


pungent, without pungency

* After Somogyi, N., Moor, A., Pek, M. (2003): The preservation and production of Capsicum in Hungary. In: De, A. K. (ed.) Capsicum. The genus Capsicum. Taylor&Francis Ltd, New York. pp. 144-162 (with permission)

2. Taltos - white fruit turning to red, indeterminate, sweet, conical, blunt, pendulous fruits, grown in the field. Fruits are 10 to 15 cm long and 5 to 6 cm in diameter. Potential yield is 30 to 35 tons/ha; 3. "Pungent apple" - white fruit turning to red, indeterminate, pungent, apple-shaped upright fruit, grown in the field. It is mainly used by the canning industry (to pickle). Fruits are 6 to 7 cm, in diameter and 4 to 5 cm long. Potential yield is 18 to 25 tons/ha; 4. Feherozon - white fruit turning red, determinate, without pungency, upright fruits, grown in the field and also under green-house conditions. Fruits are 12 to 15 cm long and 5 to 6 cm in diameter. Yield in green-house conditions is 25 to 35 tons/ha; 5. Rapires Fl - pale green turning red, indeterminate, pungent, long, conical, pendulous fruits. It can be grown in any type of controlled facilities. Fruits are 15 to 20 cm long and 3 to 4 cm in diameter. It contains Tm2 resistance. Yield depends on the technology, 7 to 8 kg/m ; 6. Tomato-shaped - green-dark green ripening to red, indeterminate, sweet, flat, round, seamed, pendulous fruits. Fruits are 8 to 12 cm in diameter and 3 to 5 cm long. The potential yield of fully ripe fruits is 18 to 20 tons/ha. 2 Paprika varieties The Hungarian condiment paprika's cultivation period is short, there are only 5 to 5 and half months available for the vegetation period. In spite of the short vegetation period, the quality of the harvested crop is excellent in most years. The high pigment and the high dry matter contents guarantee a very good base material for milling. The Hungarian varieties' yield can be up to 50% more, with improved attributes, by cultivating them in areas where the vegetation period is longer. That is based on the Hungarian-Spanish (Somogyi et al., 2003), the Hungarian-Portuguese and the Hungarian-Austrian (Derera, 2000) cooperative experiments. The full genetic potential 36

of the Hungarian cultivars is limited by the climatic conditions. Between 1993 and 2000 condiment paprika was grown on an area between 3,000 and 6,500 ha and between 26,000 and 65,000 tons of raw paprika were produced. Consequently, the quantity of the milled product varied between 3,000 and 9,400 tons. All of the condiment paprika cultivars grown in Hungary were bred in Hungary. They belong to Capsicum annuum L. covar longum by botanical classification. There are two exceptions (cv. Kalocsai A. cherry type, cv. Kalocsai M. cherry type), regarding the growth habit. They are continuous, semi-determinate and determinate. There are two types of orientation of the fruits: erect and pendulous. The categories are indicated below and the pungency is given in brackets: 1. Varieties of continuous growth habit, pendulous fruit: Szegedi 20 Szegedi 80, Szegedi 57 13, Remeny, Kalmin, Szegedi 178 (pungent), Szegedi 179 (pungent), Szegedi F-03 (pungent), Kalocsai 50, Kalocsai 90, Kalocsai V-2 (pungent), Kalocsai 15 (pungent), Csardas, Folklor; 2. Varieties of continuous growth habit, erect, Kalocsai 37 to 231; 3. Varieties of semi-determinate growth habit, pendulous fruits: Kalocsai 601, Kalocsai 702, Zuhatag; 4. Varieties of semi-determinate growth, erect fruit: Kalocsai 622, Rubin; 5. Varieties of determinate growth habit, erect fruits: Kalocsai D 601, Kalocsai D 621 (pungent); 6. The cherry-type paprika is classified as paprika, but it differs from the rest. It is Capsicum annuum covar. cerasiforme and not longum. It is pungent, its importance is found in gastronomy. If green fruit is harvested, it can be pickled or made into salad. When the ripe fruits are harvested they can be used for hot sauces or dried spice (not milled) flakes. The two cherry cultivars that differ in fruit size and growth habit are Kalocsai M and Kalocsai A. A detailed description of the most typical cultivars of each category is given below: 1. Cultivar of continuous growth habit and erect: Kalocsai 37 to 231, a sweet variety. The bush is 45 to 55 cm high, its fruits are scattered and 10 to 14 cm long, slightly bent. They are of flaming red color or dark red after post-ripening treatment. Its main value lies in its good pigment (8 to 9 g/kg) and solids content. It is a middleearly maturing variety. Its yield potential is 25-16 tons/ ha. It can be transplanted or directly seeded. It has a good tolerance to diseases; 2. Cultivar of continuous growth habit and pendulous fruit: Szegedi 80, a sweet cultivar. The fruits are 12 to 14 cm long, dark red when ripe, pigment content is 8 to 10 g/kg after post-ripening treatment. The solids content at picking is 20%. Its yield potential under intensive conditions is 20 to 25 tons/ha. It has a reasonable tolerance to diseases and can be transplanted or directly seeded. Due to an early ripeness a reasonable yield can be relied upon before the first frosts. The Szegedi F-03 is the same type but with pungency; 3. Cultivar of semi-determinate growth habit, with pendulous fruits: Kalocsai 801, a sweet variety. It has loose spreading foliage. The plant is about 40 to 45 cm long, the fruits weigh 22 to 28 g. They are straight, gradually tapering toward a pointed tip, and their color is dark red when ripe. Their pigment content at picking is 6.0 to 7.0 g/kg, going up to 8.0 to 9.0 g/kg after post-harvesting. The dry matter con37

tent is 18% when ripe. It is an early, intensive variety bringing a high yield as an exchange for watering and good nutriment supply. Its potential harvest is 20 to 22 tons/ha and it has a high tolerance to viral diseases; 4. Cultivar of semi-determinate growth habit with erect fruits: Kalocsai M 622, a sweet cultivar. The bush is 35 to 45 cm high with sparse foliage, and has a rigid stem with short internodes. Leaves are leathery and thick, so it has a good field resistance to fungal infections. The fruit is 10 to 15 cm long, gradually tapering towards a pointed tip, and is dark red when ripe. The pigment content at picking is 6.0 to 8.0 g/kg, increasing to 9.0 to 12.0 g/kg after post-harvest ripening. If transplanted, the entire crop can be harvested at the same time due to its short growing season and early, uniform ripening. It is primarily directly seeded. It is the most widely spread cultivar in the Kalocsa region. It requires intensive agronomic conditions, but the compensation is a high yield. Its yield potential is 20 to 25 tons/ha. It also has a high tolerance to diseases, which is the basis of secure production; 5. Cultivar of determinate growth habit and erect fruits: Kalocsai D 601, a sweet variety with erect fruits appearing in bunches on the stem. The bush is 30 to 35 cm high and the fruit bunches rise above the foliage and ripen uniformly. Fruits are 10 to 12 cm long, slightly bent, pointed and ripen to a deep red color. It contains 6.0 to 7.0 g/kg pigment at picking. The dry matter content is above average. It has a short growing season and early, uniform ripening. It is primarily recommended for direct seeding. On a large scale its can be harvested by machinery in one operation. The yield potential is 15 to 16 tons/ha; 6. Cherry-type paprika: Kalocsai M cherry type. It is the cultivar of pendulous fruits and loose foliage. The bush is 40-60 cm high and of continuous growth. The fruit is 3 to 3.5 cm in diameter, slightly flat and globe-shaped with a closed style point. Its surface is smooth and its pigment content is 7 to 9 g/kg. The dry matter content is 20 to 22% when ripe and the capsaicin content at picking is 120 to 140 g/100 g. It has a good tolerance against viral infections. It has a middle-early, continuous ripening and a good yield. It is primarily recommended for transplanting and requires intensive growing conditions. It needs a soil rich in organic matter that can be easily warmed up.


5. General chemical structure and composition of Capsicums

5.1. Macroscopic characteristics The dried fruits of the chilli variety are of varying sizes: 6 to 300 mm in length and 5 to 140 mm in width near the calyx. They present a variety of shapes, including the long thin conical pointed tips, short thin conical blunt tips, short broad conical to rounded tips, long broad with blunt tips and small round to oval in a few cultivars. They are laterally compressed except in the small seed-filled varieties and some paprikas. The vegetable Capsicum varieties are generally big, broad, blunt and blocky with slightly sloping sides, while the paprikas are of medium length, broad, blunt and round (Fig. I). The calyx and peduncle are usually attached, except in a few small-sized fruits and in longer varieties bred for easy destalking during harvest.


7. C a p s i c u m fruits v a r y i n g in s h a p e s a n d s i z e . 1 . tiny g l o b u l a r ; 2 . C h i l l i p i q u i n ( M e x i c o ) ; 3. M u n d u ; 4 . S i n d h o o r ; 5. L o n g R e d C a y e n n e ( U S A ) ; 6. M e x i c a n A n c h o ; 7. H u n g a r i a n p a p r i k a ; 8. B e l l C a p s i c u m (all of t h e s p e c i e s Capsicum


var. annuum,


9. t a b a s c o ( U S A ) ; 1 0 . bird's e y e ( A f r i c a n ) , ( 9 - 1 0 ) b o t h of t h e s p e c i e s Capsicum L; 1 1 . o b l o n g ; 1 2 . m e d i u m b r o a d ; 1 3 . l o n g p o i n t e d fruits of Capsicum pendulum


frutescens var.

( W i l d ) (after G o v i n d a r a j a n , V . S . , 1 9 8 5 : C a p s i c u m - P r o d u c t i o n , t e c h n o l o g y ,

chemistry, a n d q u a l i t y - Part I: History, b o t a n y , c u l t i v a t i o n , a n d p r i m a r y p r o c e s s i n g . C R C , C r i t i c a l Rev. F o o d S c i . Nutr. 2 2 : 1 0 9 - 1 7 6 ) ( w i t h p e r m i s s i o n )


The pericarp is lustrous, smooth and its color generally ranges from deep red to brownish red or orange. A few cultivars with the ripe mature color such as deep purple or white are known as also partially bleached orange color due to poor dyeing. Paprika and pimientos are generally of intense burgundy red, while vegetable Capsicums are known with both green and deep red colors at fully maturity. The fruit has two to four loci marked by longitudinal ridges with a central placenta extending from the base under the calyx to the apex end, except in vegetable Capsicums where the placenta is confined to the base (Fig. 2). Seeds are numerous, attached to the central placenta, are yellow (black in one species), smooth, 2.1 to 5 mm, discoid with thickened edge, and prominent pointed micropyle.

Fig. 2. Cross-sections of C a p s i c u m a n d c h i l l i fruits. P e : p e r i c a r p ; D : d i s s i p i g m e n t s ; S: s e e d s ; P I : p l a c e n t a (after C o v i n d a r a j a n , V . S . , 1 9 8 5 : C a p s i c u m - P r o d u c t i o n , t e c h n o l o g y , c h e m i s t r y , a n d q u a l i t y - Part I: History, botany, c u l t i v a t i o n , a n d p r i m a r y p r o c e s s i n g . C R C , C r i t i c a l Rev. F o o d S c i . Nutr. 2 2 : 1 0 9 - 1 7 6 ) ( w i t h p e r m i s s i o n )

5.2. Microscopic characteristics The microscopic characteristics of Capsicums are seen on the cross-section. A cross-section of the fruit shows the pericarp components, the cuticular layer of cells, the epidermis of the single layer of regular rectangular cells, the hypodermis of tangentially oval cells with thick walls in one or more layers, the mesocarp of parenchyma cells, having oblong, tangentially compressed and greatly varying sizes and frequently containing droplets of the red oil, occasionally crystals of calcium oxalate. In the inner regions are the vascular bundles and giant cells causing blisters on the inner surface of the pericarp, a characteristic of all Capsicum fruits, and the endocarp of rectangular cells with thick pitted walls. The diagnostic characteristics of ground Capsicum include all tissue components described above and fragments of calyx and pedicel, all mixed up. Jackson and Snowdon (1974) described the following microscopic diagnostic characteristics of Capsicum annuum L.: 40

1. numerous pale yellow fragments of the epicarp as single layer of polygonal cells with beaded walls, some showing strongly striated cuticle; 2. the abundant parenchyma of the mesocarp, thin-walled, containing orange to red oil globules and occasionally microspheroidal crystals of calcium oxalate; 3. the sclerenchymatous endocarp as groups of polygonal cells with thickened sinuous walls and having distinct pits; 4. fragments of the epidermis of the testa composed of a layer of very large lignified and markedly varying cells, unevenly thickened on the inner walls appearing as balloon-like swellings; 5. endosperm, parenchymatous cells with drops of fixed oil and aleuronic grains; 6. fragments of calyx, the inner epidermis cells with numerous trichomes with stalk and multicellular head; 7. fragments of pedicel tissues, elongated or small polygonal cells with stomata, glandular trichomes, cellulosic parenchyma, fibers in isolated groups and parenchymatous pith with large central cavity; 8. seed structures, where present, are sinuous with thin outer and thickened, pitted inner walls.

5.3. General chemical composition Carbohydrate, protein, fat and fibre are the major components which rapidly increase from the green stage to the ripe stage. Fructose, glucose, galactose and sucrose were identified by gas chromatography and enzymatical methods. Fructose is the major sugar together with glucose amounting to about 70% of total sugars. Free sugar content in seeds was reported to be higher than in the pericarp or placenta. The most fat in Capsicums is in the seed. Values for total fat vary from 9 to 16% depending on the seed content of the variety or processed sample. The high seed fat content is largely found in cultivated species. The fibre content varies greatly between the sources and reflects the amount of pedicel included in the sample. Three functionally important characteristics in the use of chillies as spice: - red color and capsaicinoids, - the pungency stimulants, - characteristic sweet aroma of paprikas and vegetable Capsicums or the pungent aroma of chillies. Other important components of Capsicums are: citric acid is a major acid component, the others being succinic, fumaric, malic and quinic acids (Table 6). Of nutritional importance is the vitamin C content of paprika, the vegetable Capsicums and chillies. Vitamin C was firstly isolated from paprika by Szent-Gyorgyi for which he was awarded the Nobel Prize in 1937. Pound for pound paprika is higher in vitamin C content than citrus fruit. Contents of vitamin C are up to 340 mg/100 g in some varieties of paprika. The content of vitamin C in dried Capsicums is only of the order of 30 to 60 mg/100 g, which indicates a large percent of loss of vitamin C in processing these Capsicums.

T a b l e 6. C h e m i c a l c o m p o s i t i o n of C a p s i c u m s * Compounds

g/100 g fresh f r u i t ( m e a n s ± S E M )


91 ±0.6


0.85 +0.1






0.81 ±0.2




0.73±0.1 mg/100 g fresh fruit (means ± S E M )

Citric acid


Fumaric acid

1.1 ±0.4

Malic acid


Oxalic acid


Quininic acid


Vitamin C


Chlorophyll a


Chrorophyll b






* After Lopez-Hernandez, J . , Oruna-Concha, M. J . , SimaLLozano,)., Vazquez-Bianco, M. E., Gonzales-Castro, M. J . (1996b): Chemical composition of Padron peppers (Capsicum annuum L.) grown in Galicia (N.W. Spain). Food Chemistry 57: 557-559 (with permission) Abbreviation: N.D. not detected

The vitamins A and B complexes also have been shown in Capsicums. Values reported of vitamin A are as high as 3315 international units in paprika and 3530 to 6165 IU in chilli varieties. The vitamin E content in fresh and dried paprika depends on the ripening stage and genetic factors. The presence of 3 to 10 mg/100 g in fresh ripe Capsicums could be an important source of vitamin E in the human diet. The loss in drying is about 5%. Oleoresin paprika had earlier been shown to have as much as 10 mg/g of a-tocopherol. A total of 14 amino acids in green Capsicum and 16 in paprika, from which asparagine and proline are dominant, were identified earlier. Recent analysis of sweet and hot varieties showed no differences in amino acids, but the composition of the seed was different from that of the pericarp. Chillies and paprika are evaluated for uniformity of shape, size, color and pungency combination of the trade types. The sweet or mildly pungent paprika is valued for its bright color and to a minor extent fot its aroma, while the chillies for their pungency and color. The paprika group contains less than 0.1% capsaicinoids, the pungency stimulants are 0 to 1.4%. In addition to water which makes up more than 90% of Capsicum fruit, fibre, pectin, glucose, starch and fructose represent the main components (Table 6).

6. Chemical taxonomy of the functional parts of the Capsicums 6.1 Capsicum: Botanical aspects Capsicum is a versatile plant used as vegetable, a pungent food additive, a colorant and a pharmaceutical. The genus Capsicum, which is commonly known as chili, "red chili", "tabasco", "paprika", "cayenne", etc., is a member of the family Solanaceae, and closely related to eggplant, potato, petunia, tomato and tobacco. Various authors ascribe 25 species to the genus, with new species to be discovered and named as the exploration of the South American tropics expands. After much work by taxonomists concerning the classification of the presently domesticated species, they have been considered to belong to one of five species, namely Capsicum annuum, Capsicum frutescens, Capsicum baccatum, Capsicum chinense and Capsicum pubescens (Bosland, 1994). The Capsicum plants grow as perennial shrubs in suitable climatic conditions. Capsicum fruits are considered to be vegetables, but botanically speaking, they are berries. Capsicum types are usually classified by fruit characteristics, i.e., pungency, color, fruit shape, as well as by their use. Capsicum species are commonly divided into two groups, pungent and non-pungent, also called hot and sweet. The history of chile peppers is one of the enthusiastic acceptances wherever they were taken. Chile is historically associated with the voyage of Columbus (Heiser, 1976). Columbus is given credit for introducing chile to Europe, and subsequently to Africa and to Asia. On his first voyage, he encountered a plant whose fruit mimicked the pungency of the black pepper (Piper nigrum L.). Columbus called it red pepper because the pods were red. The plant was not the black pepper, but a heretofore unknown plant that was later classified as Capsicum. Bolivia and Peru are the most probable ancestral home of the chile pepper, but in the world at large Mexico gets the credit for this plant, having cultivated the pepper types that entered the world's cuisines after Columbus's discovery. Capsicum species are used fresh or dried, whole or ground, and alone or in combination with other flavoring agents. Most of the Capsicum cultivars commercially cultivated in Europe and Amerika belong to the species of Capsicum annuum. However, there are a few principal types which belong to other Capsicum species. The most common types are "Bell", "Cayenne", "Cherry", "Jalapeno", "Mirasol", "Paprika", "Pasilla", "Pimento", "Serrano", "Squash" and "Wax" (all Capsicum annuum) as well as "Habanero" (Capsicum chinense), "Long Green" (Capsicum baccatum), "Manzano" (Capsicum pubescens) and "Tabasco" (Capsicum frutescens). 43

In many European languages, the name of this spice is somehow derived from the name of pepper, owing to the many confusions of pepper with other spices. A rather common designation of paprika is "sweet pepper": Spanish "pimiento dulce", French "piment doux", and Arabic "fulful halu". It should be noted that in most of these languages, the word for "pepper" may also mean "chile", so it would perhaps be more accurate to translate these names by "sweet chile" or "sweet chile pepper". Yet other tongues have names for paprika that mean "red pepper" e.g., Turkish "kirmizi biber", Bulgarian "cherven piper" and Russian "perets krasni". These may lead to confusion as in a plethora of other languages, similar names are reserved for chiles. Other English names are bell pepper and pod pepper because of the shape. Most confusingly, the English plural peppers always seem to imply some sort of paprika (also vegetable bell peppers or hot chile peppers), and never true black pepper! For reasons of clarity, the term "pepper" for Capsicum species will be avoided later in this chapter. Instead, the word "paprika" throughout for mild or medium-hot varieties and "chile" for hot varieties will be used. The word "paprika" was borrowed from Hungarian (paprika), it entered a great number of languages, in many cases probably via German. In the end, also "paprika" is derived from a name of black pepper, in this case Serbian papar. In most languages, "paprika" denotes the dried spice only, though in some (e.g., German) it is commonly used for the vegetable bell pepper. The form "paprika" is valid in countless European languages, while examples with slightly deviating spelling include Italian "paprica", Polish "papryka" and Bulgarian "piperka".

6.2. Capsicum: Chemical constituents Capsicum fruits contain coloring pigments, pungent principles, resin, protein, cellulose, pentosans, mineral elements and a small amount of volatile oil, while seeds contain fixed (non-volatile) oil. Besides these organic constituents Capsicum fruits also contain inorganic constituents, mostly potassium and sodium, calcium, phosphorus, iron, copper and manganese (Thresh, 1846; Brawer, Schoen, 1962; Brash et al., 1988; Pruthi, 2003). The pungent principles capsaicin and its structurally closely related homologes (so-called capsaicinoids) and analogues, are contained only in small amounts, as low as 0.001 to 0.005% in "mild" and 0.1% in "hot" cultivars. Apart from capsaicin, the taste of paprika is mostly due to the fixed oil which is comprised mainly of triglycerides of which linoleic, linolic, stearinic and other unsaturated fatty acids predominate. The fixed oil content of the Capsicum seeds also play an important role in the visual sensing of the paprika powder since it can dissolve and homogeneously distribute the colored substances during grinding of the dried fruits. The characteristic aroma and flavor of the fresh fruit are imparted by the volatile oil containing a range of alkylmethoxypyrazines (e.g., 2-methoxy-3-isobutyl-pyrazine, the "earthy" flavor) and a structurally diverse group (alcohols, aldehydes, ketones, carboxylic acids, and esters of carboxylic acids) of oxygenated hydrocarbons. Furthermore, the fresh ripe paprika contains sizable amounts (0.1 %) of vitamin C (ascorbic acid). It was the Hungarian biochemist Albert Szent-Gyorgyi who discov44

ered that the Hungarian paprika is a rich source of vitamin C. Later (1937) he won the Nobel Prize "for his discoveries in connection with the biological combustion processes, with special reference to vitamin C and the catalysis of fumaric acid" (Encyclopedia Britannica). During the post-ripening period and processing a substantial amount of vitamin C undergoes degradation. Paprika powder contains only a little amount of it. Paprikas derive their color in the ripe state mainly from carotenoid pigments, which range from bright red (capsanthine, capsorubine) to yellow (cucurbitene); the total carotenoid content in dried paprika is 0.1 to 0.5%. A small number of cultivars do not produce significant amounts of carotenoids; when chlorophyll levels decrease in the last stages of ripening, these chiles develop a pale hue often referred to as "white". Due to small amounts of chlorophyll and/or yellow carotenoids, the "white" is, however, more precisely described as a pale greenish-yellow. Some varieties of paprika contain pigments of anthocyanin type and develop dark purple, aubergine-colored or almost black pods; in the last stage of ripening, however, the anthocyanins get decomposed, and the unusual darkness thus gives way to normal orange or red colors. The same anthocyanins cause the dark spots which are sometimes seen on unripe fruits or particularly the stems of paprika plants and which almost all paprika varieties can develop. In other Capsicum species, anthocyanin production is a rare phenomenon.

6.2.1. Volatiles The characteristic flavor and aroma of the fresh fruits is due to their volatile oil content. The fruits of Capsicum species have a relatively low volatile oil content, ranging from about 0.1 % to 2.6% in paprika. The total volatiles are generally isolated by steam distillation. In the case of heat-sensitive compounds present, vacuum distillation-continuous solvent extraction can be used. The pure volatile oil and concentrated extracts were analyzed by GC-MS methods. Most compounds of odor significance have been tentatively identified by their mass spectra, and the identification was confirmed by checking the retention time and mass spectra of authentic reference compounds. When Buttery et al. (1969a,b) identified 3-isobutyl-2-methoxypyrazine (1) (Fig. 3) as a characteristic aroma compound, the alkyl-methoxypyrazines aroused great interest among flavor chemists. The alkyl-methoxypyrazines have been shown to be widely distributed in vegetables and with a greenish sweet smell that possibly plays a significant role in the aroma of salad vegetables (Murray, Whitfield, 1975). In the volatiles of the green bell fruits (C. annuum var. grossum), different Capsicum fruits with strong odor, the major character-determining component 3-isobutyl-2-methoxypyrazine (1) was easily identified by GC-MS technique and sensory analysis. Other odor impact compounds identified of the bell capsicum volatiles were hexanal (2), which has a mild nutty odor, cis-3-hexen-l-ol (3) with a green leaf odor, the C(9) unsaturated ketones (non-l-en-4-one, non-trans-2-en-4-one, non-2trans-5-trans-dien-4-one), nona- and decadienals (non-2-trans-6-cis-dienal, non-2trans-4-trans-dienal), and a few terpenic compounds (e.g., limonene, trans-betaocimene, linalool (4) (Buttery, et al., (1969a) (Fig. 3). 45


./CH X






C H ,— C H — O 1 1 — CI I C I





CH — C H 3



/ H




/ —





C H — C H — OH 2




-CH — C H = C H - C - C H 2






°,x 3

CH —CH 3




CH —CH —O / Af-[(4-Hydroxy-3-methoxy-phenyl) methyl]-8-methyl. (£)-8-Methyl- Af-vanillyl-6-nonenamide, (Ref. USP, Ref. 6.2.3).} In the formulation we used USP quality of capsaicin ("capsaicin natural", CAS Number: 404-86-4), since PH EUR does not involve capsaicin paragraph (USP 27NF 22, THE UNITED STATES PHARMACOPEIAL Convention, INC. 12601 Twinbrook Parkway, Rockwille , 2003). API supplier was audited and they released a copy of PMF for capsaicin. The pharmaceutical characteristics of the substance: Properties: - Characteristics: - pale brownish-yellow, amorphous powder - intensive and irritating odor even in very low air-volume concentration - Morphology: - particle size is not homogeneous, it is varying in a wide range between 0.05-2 mm (like a usual vacuum-dried extract) - Polymorphism: - no polymorphism of the substance is known - Solubility: - good in methanol, ethanol and acetone, practically insoluble in water Chemical properties: testing and specification is upon USP 83/2001 EC Guide (Directive of the European Parliament and of the Council on the Community code relating to medicinal products for human use) (Handbook of Pharmaceutical Excipients 5th Edition, edited by Raymond Rowe, Paul Sheskey and Paul Weller, USA, October 2005).

160 Excipients' quality of P H EUR requirements Excipients' quality was meeting PH EUR requirements, involving the BSE/TSE aspect, when relevant, as well. (Directive of the European Parliament and of the Council on the Community code relating to medicinal products for human use (2001) (83/2001 EC Guide). Excipients used in formulation - as above -were expected to be chemically - indifferent, - to serve the solid dosage form requirement (Handbook of Pharmaceutical Excipients 5th Edition (2005): Eds: Rowe, R., Sheskey, P. and Weller, P., USA, October 2005) - not to involve lactose in order of protection of lactose-sensitive patients As from view of formulation, the excipients must have proper physical-, physicalchemical characteristics for powder rheology in order to assure good homogenity, fluidity and lubrication for compression. Coating powder mixture quality Coating powder mixture quality is by IHS, formulated for serving the above-mentioned purposes. Its red color was aiming at some association with the Hungarian paprika. Additionally, for suspension phase or solvent of capsaicin purified water was used exclusively in order to decrease the risk of organic residual.

11.2.3. Packaging Details of the packaging are shown in Table 33. T a b l e 3 3 . S u m m a r y of a s p e c t s of p a c k a g i n g yes


1. Primary blister



C o l d blister

vial 2. Secondary carton box


p a c k a g e insert booklet

3. D e s i g n design i n v o l v i n g b l i n d - c o d i n g

* abbreviations: PVC, Polyvinyl Chloride PVdC, Polyvinylidene Chloride ALU, Aluminium


A good packaging retains the products' quality, is easy to handle by patients and externally serves some aesthetic aspects. Blister packaging has been selected, because it was an individual packaging by dosage form, and the indicated packaging materials were resistant against humidity, thus supporting the stability of the tablet.

11.3. Formulation 11.3.1. Formulation steps In the first step a lab-scale batch (appr. 3 kg) was planned to be produced, in order to confirm the planned composition, then a pilot batch (100 000 tablets) was targeted. As we have worked out the process in detail, FP Specification has been set up after the complex testing of the tablets.

11.3.2. FP Specification The details of the FP are listed in Table 34. T a b l e 3 4 . F i n i s h e d p r o d u c t ( F P ) of t h e s p e c i f i c a t i o n s a r e s u m m a r i z e d in this T a b l e . * Aspect




R e d c o l o r e d , b i c o n v e x f i l m tablet w i t h s m o o t h surface

Dosage form

- Dimensions: - height - diameter - shape - average weight - individual weight - disintegration time - friability -CU


Practical average ± 1 0 % (mm) 10 ± 3 %


biconvex T h e o r e t i c a l a v e r a g e ± 1 0 % (g) P r a c t i c a l a v e r a g e ± 1 0 % (g) 30 min. NMT 1 % 85-115%

- Identity: -API


- binder


- lubricant - glidant

positive positive

- assay: - capsaicin I m p u r i t y test


- RS

Total R S N M T

- Microbiology

III.A. - P H

* see Table 30, for other additional explanation




11.3.3. Samples for clinical trial After manufacturing and testing the samples taken from the pilot batch and evaluation of the BMR, Master Batch Documents and Master Quality Documents have been created for approval of QA. All the other SOPs, safety rules involving any other aspect of producing a clinical sample have been created formerly and authorized to achieve quality assurance requirements as well. Then, a new pilot batch was planned to be produced for two purposes: - t o take samples for clinical trial ( - to take samples from the same batch for stability testing of the formulation ( Preparation of samples for clinical trial Samples for clinical trial were planned to be taken after the release of the batch. The number of the samples was calculated on the basis of the Clinical Protocol, their packaging and coding were compliant with GMP regulation (Eudralex Volume 4 (2007): Good Manufacturing Practice, European Comission). Exposing, stability and packaging of the planned final product Samples for exposing and stability testing were packed in the same packaging as the planned final packaging. The product's stability is a main quality requirement. It means that the product packed in its final form retains chemical, pharmaceutical stability and pharmacological activity within a certain range which was set up in the finished product specification. (If there is any reasonable reason, some requirements of the release and expiry specification may be different within a certain range.) A complex stability project has been created for exposition and testing by ICH Guides in order to determine expiry specification and expiry date of the product as well (International Conference on Harmonization (ICH) (2003) (Ql, Q8), Brussels).

11.4. Plant batch manufacturing and process validation Scaling up for plant batch is almost the final step of a development project. It was planned and achieved by the collected information and data of the former steps. On the other hand, planning of the process involves the site manufacturing machinery and technical supplying system too. Taking into account all of the above, batch size could be determined easily. After plant batch manufacturing and quality control of the taken samples a final report evaluates the conclusions of scaling up. Process validation and cleaning validation must be achieved upon written plans, then evaluated in a written report for 163

justification of the process and its parameters. Validation batches are also involved in stability project. Cleaning validation is the last important plant activity belonging to the validation procedures, while certifying the proper method of cleaning after operation. Approved validation reports usually confirm not only the processes, but the MBDs and MQDs belonging to the as well.

1 1 . 5 . Summary of the Chemical-Pharmaceutical development The chemical-pharmaceutical part of the products' dossier is one of the main 3 detailed sections. This is for certification and documentation of all activities of the product development from pre-formulation to plant batch production on the basic principle of QUALITY. For certification of the above, we assume the next points to be relevant: - written plans are required (involving formulation and processes) for the different steps of development, - standard quality and test methods of the used starting materials are required, - standard methods are required for quality control of products at different levels of development, - development activity must follow QA rules, especially in documentation: - basic documents must have stability control, - plans must be performed, and evaluated in correct reports, - consecutive steps must follow an approved report and conclusion of the former step, - records must be controlled and approved, - the plant batch size production and the process belonging to it must be validated, - a required number of batches must be subject to stability testing, - in the final evaluation, i.e. in the Chemical and pharmaceutical expert report, expiry date, storing condition and other notes must be clarified and declared unambiguously. The planned appearance of capsaicinoids containing pills is seen in Fig. 31.

Fig. 31. T h e v i s u a l c h a r a c t e r i z a t i o n of c a p s a i c i n o i d s c o n t a i n i n g pills (see t h e p i c t u r e in c o l o r s o n p. 2 5 5 )


12. Clinical pharmacological studies with capsaicinoids alone and with combination of capsaicinoids with nonsteroidal anti-inflammatory drugs 12.1. Main aims of clinical pharmacology and its relation to the evidence-based medicine The evidence-based medicine (EBM) was established as the basis of the activity of medical treatment, in the everyday medical practice. The general aims of this EBM are to give exact and clinically well proved evidences for making correct diagnoses and for the treatment of patients suffering from different diseases. In the last 3-4 decades (since 1970) only the "problem orientated medicine (POM)" has been emphasized in the graduate and postgraduate medical education. This internationally well accepted trend has been applied in the medical education and in the diagnostics of diseases in patients. The final aims of our medical activity are to give scientifically well proven medical treatments (including the surgical treatment, treatment with different drugs, etc.) for the patients. The drugs have a key role in the medical treatment. It was earlier suggested that the medical treatment can be based on the: 1. classical "old" medical experiments (e.g. Chinese medicine) and 2. classical (theoretically well planned) pharmacological research. The pharmacological studies were (and are) based on the results obtained from: 1. various animal experiments; 2. results obtained on the human isolated cell cultures [especially from that time, when the Leagu(ag)e for Animal Defense (LAD) emphasized the defense of animals to be involved into the research activity]; and 3. human pharmacological studies. The animal observations played essential roles in the discoveries of different physiological and pharmacological facts and events. The researchers used mice, rats, guinea pigs, dogs, chicken, etc. in their pharmacological research. Probably mice and rats are the most accepted animals for physiological and pharmacological studies. The different human cell cultures were also involved in medical research to clear up the exact details of different physiological and pharmacological mechanisms of different compounds. There is, however, a significant problem with the exclusive use of the isolated human cell lines for the "human medical research", because these cell lines have no innervation, no correct hormonal regulation, etc. Of course, the results of these types of observations are basically necessary to establish the human pharmacotherapy. Without denying the importance of these types of research on the animal observations and on isolated human cell line, the most important physiologi165

cal and pharmacological regulatory mechanisms remain to be unclear in the human body. The results of the research on cell cultures cannot be adapted directly in the production of antibodies against the suggested mediators, in patients with inflammatory bowel diseases. The human pharmacotherapy has undergone significant changes in the last decades. Earlier the efficacy of drug therapy was based only on the observation made by the physicians. These observations can be done retrospectively and prospectively (e.g. the efficacy of antituberculotic treatment was easily acceptable in Hungary, because earlier we had no good antituberculotic drugs, such as Streptomycin; isonicotinic hydrazine, INH; rifampicin, Tubocin®, etc). The Hungarian people died before the discoveries of these drugs, however, the Hungarian patients were healed (practically in their whole number) after the introduction of these drugs in the everyday medical treatment. Consequently the "Hungarian Disease" ("Morbus Hungaricus") disappeared in the last decades. This process was a clear medical proof for the efficacy of medical treatment in patients. Penicillin was accidentally discovered by Fleming (1922). Its efficacy could be proved clearly in patients with different bacterial infections (the fever disappeared and the patients got better in a short time after the introduction of penicillin). A. Fleming received Nobel prize in 1945. When the different drugs are used in the treatment the patients with different diseases, the efficacies of different drugs can be evaluated by parametric (measureable) and nonparametric (subjective) parameters, which change from time to time. Clinical pharmacology was established in the years of 1960 over the World (at the same time in Hungary). Professor Tibor Javor (Second Department of Medicine, Medical University of Debrecen, Hungary) and his research group (the writer of this chapter personally participated in the establishment of clinical pharmacology of anticholinergic drugs) were pioneers in the establishment of human clinical pharmacology in Hungary. The results of the human clinical pharmacology offered the most principal research argument for the "classical human pharmacology", e.g. medical treatments. The main aims of the human clinical pharmacology are: 1. To give objective data on drug absorption from the gastrointestinal tract (in case of orally applicable drugs), metabolism in the human body (dominantly by the liver) and excretion of drugs (or their metabolites) by the urine or by the stool; 2. To observe the main pharmacodynamic actions of drugs; 3. To measure the drug (or metabolites) in the serum, urine, stool, after the application of the drug given in different doses; 4. To identify the oral and parenteral dose rate of the drug; 5. To identify the correlation between the pharmacodynamic actions and pharmacokinetic parameters in humans; 6. To identify the similarities (and differences) of these parameters obtained in healthy human persons (volunteers) and patients; 7. To examine these parameters before and after chronic treatment with drug(s); 8. To find a correlation between the results of the clinical pharmacological studies and those of classical human pharmacological treatment (which finally represent the everyday medical treatments). 166

Clinical pharmacology represents a classical field in the medical sciences. It involves a classical multidisciplinary knowledge, which can be given only by experts working in the different fields of medical sciences (chemists, mathematicians, physicians, drug technologists, drug industrial experts, clinical pharmacologists, etc.). Of course, the special laboratories and medical wards (clinical pharmacological units) are basically necessary for carrying out these studies. There is no doubt about that the human clinical pharmacology has developed extremely in the last decades in the World (including Hungary). The classical clinical pharmacology plays an essential role in the innovative drug research. Furthermore, the innovative drug research and clinical pharmacology can be evaluated by the internationally accepted parameters (it was true already before Hungary joined the European Union, 2004).

12.2. Our special scientific problems in the human clinical pharmacology of capsaicinoids alone and together with the application of aspirin, diclofenac and Naproxen We mentioned earlier that "capsaicin" (which name is only used in the classical physiological and pharmacological research) does not represent only one chemical structure. The capsaicin of plant origin consists of capsaicin, dihydrocapsaicin, norcapsaicin and nordihydrocapsaicin as main components. Capsaicin is transformed by the metabolism in human and animals into dihydrocapsaicin. Physiological and pharmacological studies indicated clearly that there is no difference between the chemically very similar capsaicinoids in animal experiments (Buck, Burks, 1986). We had the following main scientific problems: 1. There was as important question as to how many components can be put into the planned drug products (capsaicinoids + aspirin; capsaicinoids + diclofenac and capsaicinoids + Naproxen). The measured parameters (chemical measurements of the chemical compound in human biological samples) in the human pharmacological studies inform us on the efficacy of the drugs or drug combinations from the point of clinical pharmacology. Capsaicin and dihydrocapsaicin give the most important part of the chemical components of "classical capsaicin preparate". According to special consultations with the experts of the Hungarian Institute of Pharmacy, capsaicin and dihydrocapsaicin can be measured as main components in the human clinical pharmacological studies. Furthermore, dihydrocapsaicin is the main metabolite of capsaicin. Consequently the measurements of dihydrocapsaicin can be used as one of the main chemical components for the pharmacokinetic approach to capsaicinoids in the different human clinical pharmacological studies; 2. When we tried to establish new drug combinations (capsaicinoids + aspirin; capsaicinoids + diclofenac and capsaicinoids + Naproxen) we had to measure two chemical compounds from the two main components of capsaicinoids (capsaicin and dihydrocapsaicin) during the clinical pharmacological studies (together with aspirin, diclofenac and Naproxen ) from the human sera; 167

3. The acceptance of this standpoint resulted very important consequences for our clinical pharmacological studies: - the pharmacokinetic measurements of capsaicinoids during the pharmacokinetic studies in healthy human subjects and in patients with different disorders (absorption rate, metabolisation, excretion of capsaicinoids) can be characterized by the results of measurements of capsaicin and dihydrocapsacin, - i f we do not have to measure all capsaicinoid components (having the same physiological mechanisms) the carrying out of these studies becomes considerably cheaper; 4. We have to perform "classical pharmacodynamic" and "pharmacokinetic" studies with capsaicinoids alone in healthy human subjects, because there have been no similar observations published in the world literature up to now (either in healthy human subjects or in patients with different disorders); 5. Since we will use different combinations of capsaicinoids plus NSAIDs (aspirin, diclofenac, Naproxen), consequently we have to carry out the human Phase I study with capsaicinoids alone and together with aspirin, diclofenac and Naproxen.

12.3. Principal schedules for the human Phase l-ll studies with capsaicinoids alone and together with aspirin, diclofenac and Naproxen 12.3.1. Preparation of protocols for the human clinical pharmacological studies (including Phase I to IV) Medical points of the preparation of the study protocols Very carefully carried out preparation of the medical protocols is the basis for prospective, randomized and multicentric studies (including one or more countries of the World). These protocols describe the medical points of clinical pharmacological studies [clear identification of the aim(s) of the study(studies) in the healthy human subjects or patients with different diseases involved in the study, their exact inclusion and exclusion criteria, schedule of the well-planned examinations, clinical controlling examinations and their time, drop-out, determination of end points of the studies]. The protocols prepared for the clinical study (studies) have to be accepted by nationally or internationally well known experts only from the medical points of views. The place and the names of person(s) responsible for performing these clinical pharmacological studies should be given. Details of the whole documentation of the clinical pharmacological studies (informed consent, health insurance, case report sheet, circumstances of the data, preparation for the chemical pharmacological studies, storage, transfer of biological samples, the registration and archivation of the obtained results and finally to write a 168

final summary of the whole clinical pharmacological study) are clearly indicated and regulated. The final medical control is given by the National Institute of Pharmacy, and this Department gives the final permission (from the medical points of views) for carrying out the studies.

12.3.2. Control of the protocols by the National or Regional Clinical Pharmacological and Ethical Committees After the acceptance of different protocols by the National Institute of Pharmacy (including the necessary preliminary decisions by the national and internationally well known experts), the protocols will be controlled by the National Clinical Pharmacological and Ethical Committee of Hungary (in case of Phase I to III, and in some extent in case of Phase IV studies) from the ethical points of views, looking for the health and juristic defense of participants involved in the different studies. These Committees include medical experts from the different fields of medical sciences, lawyers, ethical experts, and nurses. If the National Clinical Pharmacological and Ethical Committee of Hungary will accept the correct circumstances (including the medical and ethical aspects of the study planned to be carried out) of the planned clinical pharmacological study (studies), then the examinations can be commenced.

12.3.3. Pharmacokinetic and pharmacodynamic effects of capsaicinoids only Human Phase I clinical pharmacological study The results of different animal experiments clearly demonstrated that the pharmacodynamic effects of capsaicinoids depend on the applied doses. Four different doses of capsaicinoids are able to produce four different pharmacological effects (Szolcsanyi, 1996; Mozsik et al., 2000): 1. small doses of capsaicinoids stimulate the capsaicin-sensitive afferent nerves (these actions are reversible ones); 2. small doses of capsaicinoids, but in a little higher than those mentioned above, produce the inhibition of the same capsaicin sensitive afferent nerve fibres (which is also a reversible process); 3. higher doses of capsaicinoids (than those mentioned under point 2) produce an injury of the capsaicin sensitive afferent nerves (which is probably a reversible one); 4. the extremely high doses of capsaicinoids produce an irreversible injury of these nerves. These pharmacological studies were carried out in animal experiments. In healthy human subjects we applied the very small doses of capsaicinoids (Mozsik et al., 169

2005a), which stimulate the capsaicin-sensitive afferent nerves. The capsaicinoidsinduced side effects (diarrhoea, development of malignant diseases) were observed and published in the world literature, in connection with the administration of extremely high doses of capsaicinoids. We have to emphasize that the capsaicinoids were applied in different capsaicin extracts in these studies (and not in a chemically pure form). The pharmacokinetic and pharmacodynamic Phase I observations were planned to be carried out only with small doses of capsaicinoids (up to 1.0 to 1.2 mg/person/day). The primary aims of these studies are to identify the tolerability, safety and pharmacodynamic aspects of capsaicinoids in healthy human subjects and in patients with different diseases. The capsaicinoids are orally given in doses 400 to 800 pg/person/day. Human clinical pharmacological Phase I study with capsaicinoids plus nonsteroidal antiinflammatory drugs in healthy human subjects Human clinical pharmacological phase I study with capsaicinoids plus aspirin

The dose of aspirin is 100 mg/day/person, in case of aspirin resistance 300 mg/day/person, respectively, estimated by many international studies, and these doses have been accepted recently by the European and American Societies of Cardiology (2006) in patients with myocardial infarction. Consequently the doses of aspirin are internationally well established. We planned to carry out human Phase I studies with three arms: 1. E D dose of capsaicinoids (400 pg in oral dose); 2. aspirin (100 mg and 300 mg), given orally; 3. aspirin (in doses mentioned above) plus capsaicinoids (ED ), given orally. The observations will be carried out in healthy human subjects (volunteers). Every subject will undergo the different observations, however, the actually applied dose will be selected according to randomization. 50

50 Human clinical pharmacological Phase I study with capsaicinoids plus diclofenac

We planned also a human clinical pharmacological phase I study with capsaicinoids plus diclofenac with three arms: 1. capsaicinoids (in dose of E D ) , (400 pg oral dose); 2. diclofenac (25, 50 and 75 mg), given orally; 3. diclofenac (25, 50 and 75 mg) plus capsaicinoids (in dose of ED ), given orally. The clinical pharmacological observations will be done in healthy persons (volunteers). 500


170 Human clinical pharmacological Phase I study with capsaicinoids plus Naproxen

Healthy human subjects (volunteers) are included in these studies. The persons receive all the treatments mentioned below: 1. capsaicinoids in dose of E D (400 pg oral dose); 2. Naproxen (250, 375 and 500 mg), given orally alone; 3. capsaicinoids (in dose of ED ) plus Naproxen (in the above-mentioned doses) given together orally. 50

50 Human clinical pharmacological Phase II studies in patients Human clinical pharmacological Phase II study in patients with thromboembolic diseases (myocardial infarction, stroke, thromboembolic events)

The patients selected for the study were treated with aspirin (in doses of 100 or 300 mg given orally) without and with co-administration of capsaicinoids (orally given ED ). The general laboratory parameters, ECC, platelet aggregation were registered dayly, together with the subjective complaints of patients. Of course, the pharmacodynamic examinations were also carried out in the patients (treated with capsaicinoids alone, aspirin alone and capsaicinoids plus aspirin). 50 Human clinical pharmacological Phase II studies in patients with different degenerative locomotor diseases

Similar types of the human Phase II examinations are carried out in these patients as those mentioned in Section These laboratory examinations (plus others related to the degenerative locomotor system) and semi-quantitative parameters (complaints of patients) were registered according to the protocols. The patients were treated with diclofenac (25, 50 and 75 mg given orally), capsaicinoids (given orally in doses of E D ) and diclofenac (in the above-mentioned doses) plus capsaicinoids (in dose of ED ) given orally. Another group of patients received Naproxen (in doses of 250, 375 and 500 mg orally) instead of diclofenac. The further details of this study were the same as in the case of diclofenac. The pharmacodynamic examinations were carried out during the clinical observation. 50



Note We received absolutely new information from the chronic toxicological studies in Beagle dogs (2008). These animals were treated with different doses (0.1, 0.3 and 0.9 mg/kg bw/day orally given) of capsaicin(s) for one month. No toxicological side effects were observed in these dogs during the whole treatment periods. We noticed surprisingly that capsaicin (capsaicin and dihydrocapsaicin) could not be detected in the sera of Beagle dogs, either by High Pressure Chromatography (HPLC)* or by Liquid Chromatography-Mass Spectrometry (LC-MS)** at any time after the oral application of capsaicin (in doses of 0.1; 0.3; 0.9 mg/kg bw/day). The limit of detection by HPLC is 20 nanogram/ml serum and its value is 26 fg for capsaicin and 20 fg for dihydrocapsaicin by LC-MS. These results suggest that we will not be able to produce a classical pharmacokinetic study for capsaicin and dihydrocapsaicin in healthy human subjects and in patients with different diseases because the dose range of capsaicin is 0.4-1.2 (400-1200 microgram)/ adult persons/ day.

* Mozsik, Gy., Past, T , Perjesi, P., Szolcsanyi, J.: Determination of capsaicin and dihydrocapsaicin content of dog's plasma by HPLC-FLD method. In: Mozsik, Gy., Past, T., Perjesi, P., Szolcsanyi, J.: Original Reports on Toxicology of Capsaicin VII. 8-Day Oral Toxicity Study of Test Item Capsaicin Natural USP 27 in Beagle Dogs (Final Report). LAB International Research Centre Hungary Ltd. Veszprem by the date of final report 13 June 2008. Study Code: 07/496-100K pp. 1-35 in text and 190 pages in Appendices. (Appendix 2.11) pp. 1-37 (2008) ** Boros, B., Dornyei, A., Felinger, A.: Determination of capsaicin and dihydrocapsaicin in dog plasma by Liquid Chromatography-Mass Spectrometry (Analytical method report) PTE TTK Analitikai Kemiai Tanszek, Pecs, Hungary (2008) 172


The study was supported by the grant of the National Office for Research and Technology, "Pazmany Peter program" (RET-II 08/2005). The authors express their sincere thanks to Mrs. Judit Szabo for her excellent help in the preparation of this monograph. They are also very grateful to Akademiai Kiado (Budapest, Hungary), especially to Mrs. Judit Kerpel-Fronius for her precious corrections during the copy editing and proofreading process.



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1. European Commission (Health and Consumer Protection Directorate General Directorate-C Scientific Opinion): C2 Management of Scientific Committees II Scientific Cooperation and Networks ( 1.1 Opinion of the Scientific Committee on food on capsaicin Terms of reference The Committee is asked to advise the Commission on substances used as flavoring substances or present in flavorings or present in other food ingredients with flavoring properties for which existing toxicological data indicate that restrictions of use or presence might be necessary to ensure safety for human health. In particular, the Committee is asked to advise the Commission on the implications for human health of capsaicin in the diet. Introduction Previous evaluations The Committee of Experts on Flavouring Substances of the Council of Europe evaluated the capsaicinoids in Capsicum preparations used as flavourings. A TDI of 0-0.2 mg/kg bw, expressed as total capsaicinoids, was numerically derived from the results of a population based case-control study conducted in Mexico City, where chilli pepper consumers were at high risk for gastric cancer compared with non-consumers. The daily intake of the chilli pepper consumers was estimated to be 4 mg capsaicinoids/kg bw and a safety factor of 20 was applied. In addition, general limits of 5 ppm for foods and beverages, 10 ppm for hot foods and beverages, 20 ppm for hot ketchup and 50 ppm for tabasco, harissa, hot pimento oils and similar preparations expressed as total capsaicinoids were suggested (Council of Europe, 2001). Current regulatory status Capsaicin is listed in the register of chemically defined flavouring substances laid down in Commission Decision 1999/217/EC (EC, 1999), as last amended by Commission Decision 2002/113/EC (EC, 2002). 207

Chemical characterisation Name: Capsaicin (N-(4-hydroxy-3-methoxybenzyl)-8-methyl-trans-6-nonenamide) Capsaicin in Capsicum preparations is always accompanied by other capsaicinoids: mainly dihydrocapsaicin, but also small amounts of nordihydro-, homo-, homodihydro-, nor-, and nornorcapsaicin. The capsaicinoids present in the Capsicum fruit are predominantly capsaicin and dihydrocapsaicin, making up 80 to 90%. The ratio of capsaicin to dihydrocapsaicin is generally around 1:1 and 2:1 (Govindarajan, Sathyanarayana, 1991). Synonyms: 8-Methylnon-6-enoyl-4-hydroxy-3-methoxybenzylamide; trans-8 methyl-N-vanillyl-6-nonenamide; Isodecenoic acid vanillylamide (Fig. 32) FL No: 16.014 CAS No: 404-86-4 FEMANo: 3404 CoENo: 2299 EINECS: 206-969-8 Structure:

Fig. 32.

8-Methylnon-6-enoyl-4-hydroxy-3-methoxybenzylamide; trans-8-methyl-N-vanillyl-6-nonenamide; Isodecenoic acid v a n i l l y l a m i d e

Exposure assessment Capsaicinoids are mainly ingested as naturally occurring pungency-producing components of capsicum spices (chilli, cayenne pepper, red pepper). They typically range from 0.1 mg/g in chilli pepper to 2.5 mg/g in red pepper and 60 mg/g in oleoresin red pepper (Parrish, 1996). Pepper varieties from Capsicum frutescens, annuum and chinense were found to contain 0.22-20 mg total capsaicinoids/g of dry weight (Thomas et al., 1998). Cayenne pepper samples had mean capsaicin and dihydrocapsaicin contents of 1.32 and 0.83 mg/g dry weight, respectively (Lopez-Hernandez, 1996). The consumption of capsicum spices was reported to be 2.5 g/person/day in India, 5 g/person/day in Thailand (Monsereenusorn, 1983; Monsereenusorn et al., 1982) and 20 g/person (one chilli pepper) per day in Mexico (Lopez-Carrillo, 1994). Assuming a content of capsaicinoids in these spices of about 1%, the daily intake of capsaicinoids in these countries has been estimated to be 25-200 mg/person/day or in the case of a person with 50 kg body weight 0.5-4 mg/kg bw/day (Council of Europe, 2001). The maximum daily intake of capsaicin in the U.S. and Europe from mild chillies and paprika was roughly estimated to be 0.025 mg/kg bw (Govindarajan, Sathyanarayana, 1991), equivalent to 1.5 mg/person/day. According to a recent estima208

tion, the mean and maximum intake of capsaicin from industrially prepared food products containing the recommended general limit of 5 pm would be 0.77 and 2.64 mg/day, respectively (CREDOC/OCA, 1998). Hazard identification/characterisation Capsaicin and other members of the group of capsaicinoids produce a large number of physiological and pharmacological effects such as effects on the gastrointestinal tract, the cardiovascular and respiratory system as well as the sensory and thermoregulation system. These effects result principally from the specific action of capsaicinoids on primary afferent neurons of the C-fiber type. This provides the rationale for their use to treat some peripheral painful states, such as rheumatoid arthritis (Surh, Lee, 1995) In addition, capsaicinoids are powerful irritants, causing burn and pain at low concentrations on the skin and mucous membranes. Given orally, they induce an increase of salivation and gastric secretion, a rapid change of sensation, warm to intolerable burning, and gastrointestinal disorders depending on the dose (Govindarajan, Sathyanarayana, 1991). Absorption, distribution, metabolism and excretion Capsaicinoids, when administered to rats intragastrically are readily absorbed and metabolized to a great extent in the liver before reaching the general circulation and extrahepatic organs (Donnerer et al., 1990). In vitro and in vivo studies have shown that capsaicinoids are metabolized by different pathways: (1) hydrolysis of the acidamide-bond and oxidative deamination of the formed vanillylamine, (2) hydroxylation of the vanillyl ring, possibly via epoxidation, (3) one electron oxidation of the ring hydroxyl forming phenoxy radicals and capsaicinoid dimers, (4) oxidation at the terminal carbon of the side chain (Surh, Lee, 1995). Within 48 hrs after oral administration of dihydrocapsaicin to male rats, 8.7% of the dose were excreted unchanged in urine and 10% in faeces. Metabolites found in urine were vanillylamine (4.7%), vanillin (4.6%) vanillyl alcohol (37.6%) and vanillic acid (19.2%) in free form or as glucuronides (Kawada, Iwai, 1985). Based on results of Miller et al. (1983), who demonstrated the covalent binding of dihydrocapsaicin to hepatic microsomal proteins, the formation of electrophilic intermediates (arene epoxides, phenoxy radicals or quinone type derivatives formed after O-demethylation) and subsequent covalent binding to cellular macromolecules is discussed to play a role in the etiology of capsaicin-induced toxicity including mutagenicity and carcinogenicity (Surh, Lee, 1995). Acute toxicity The acute toxicity of capsaicin shows a large variation depending on the route of administration. In male mice, the LD 50 varies from 0.56 mg/kg bw (i.v.) to 60-75 mg/kg bw (in ethanol) and 190 (122-294) mg/kg bw (in dimethyl sulfoxide), following intragastric intubation. The possible cause of death was considered to be due to respiratory paralysis (Glinsukon et al. 1980). Intraduodenal and intragastric administration of 10% Capsicum as well as 0.014% capsaicin in 0.85% saline to male rats produced morphological damages in the duodenal mucosa (Nopanitaya, Nye, 1974). 209

Subacute/subchronic toxicity A 4-week feeding study with groups of 5 male B6C3F1 mice with 0, 0.5, 1.0, 2.5, 5.0, 7.5, and 10% ground red chilli (Capsicum annuum) in the diet showed slight glycogen depletion and anisocytosis of hepatocytes in the 10% group. Other organs did not reveal any lesions. General health, body weight and food intake were not adversely affected (Jang, Kim 1988; Jang et al., 1992). Groups of 10-14 rats were fed by stomach tube with 50 mg/kg bw/day capsaicin or 0.5 g/kg bw/day Capsicum extract for 10-60 days. There were significant reductions of growth, plasma urea, glucose, phospholipids, triglycerides, total cholesterol, free fatty acids, glutamic pyruvic transaminase, and alkaline phosphatase in both groups with a tendency for Capsicum treated animals to show more adverse effects. No gross pathological changes and no differences in organ weights from control values were observed at autopsy, only a slight hyperemia in the livers and reddening with increasing mucous materials in the gastric mucosa. The organs, however, were not examined histopathologically (Monsereenusorn, 1983). BALB/c mice received an alcoholic chilli extract in drinking water 5 days a week till 16 months of age (27 males, 25 pg capsaicin/week, equivalent to about 0.125 mg/kg bw/d) or on the tongue 2 days a week for 14 months (22 males without and 19 males with 1% atropin solution prior to application, 50 pg capsaicin/week, equivalent to about 0.25 mg/kg bw/d). Compared with 40 untreated mice, the treated animals showed increased mortality and histopathological changes in liver, kidneys, stomach and tongue. The lesions in the liver observed in all treated mice were in the form of focal necrosis with inflammatory cells around, fatty changes and fibrosis (Agrawal, Bhide, 1987). 36 male Syrian hamsters received 20 pi alcoholic chilli extract with 50 pg capsaicin (equivalent to about 0.5 mg/kg bw/d) 5 days a week by cheek pouch application for 14 months. 30 untreated animals and 17 hamsters treated with 20 pi alcohol were used as controls. The animals treated with chilli extract had increased mortality and histopathological lesions in liver, kidneys, stomach and cheek pouch. The main lesions were liver cirrhosis, observed in 49 % of examined livers from exposed hamsters compared to 8 and 17 % in the control groups and glomeruli degeneration in 50% of examined kidneys of exposed animals compared to 8 and 0 % in the control groups, respectively (Agrawal, Bhide, 1988). Capsaicin, administered intraperitoneally to adult male mice at doses of 0.4, 0.8 or 1.6 mg/kg bw/day on 5 consecutive days, did not induce significant alterations in epididymal weights, caudal sperm counts, testicular weights or testicular histology. In the sperm morphology assay, sperms at 1, 3, 5 and 7 weeks did not reveal any treatment-related increase in the incidence of sperm-head abnormalities (Muralidhara, Narasimhamurthy, 1988). In a 13-week study performed to determine the maximum tolerated dose, groups of 10 male and 10 female B6C3F1 mice received a mixture of 64.5% capsaicin and 32.6% dihydrocapsaicin at those levels of 0, 0.0625, 0.125, 0.25, 0.5, and 1% in the diet. Significant reduction of food intake and body weight gain in all dose groups, especially in treated females, and significantly increased liver/body weight ratios of both sexes and renal toxicity (focal tubular dilatation) in the 1 % treated males were observed (Akagi et al., 1998). 210

Capsaicin (purum) was administered at concentrations of 0.0625, 0.125, 0.25, 0.5 and 1% in the diet of groups of 4 male and 4 female Swiss albino mice for 35 days. When the animals died at an age of 62-126 weeks, one adenocarcinoma of the duodenum had developed at each dose level, except for the highest dose, while no such tumours occurred in a historical control group of 100 males and 100 females. There was no concurrent control group and the observed tumour incidence was not doserelated (Toth, Gannett, 1992; Toth et al., 1984). Chronic toxicity/carcinogenicity 15 out of 26 rats fed for seven months with 10% chillies in a semisynthetic diet containing ardein, a purified protein of the ground nut, developed neoplastic changes in the liver (hepatomas, multiple cystic cholangiomas, solid adenomas or adenocarcinomas of the bile duct). Although no tumour developed in rats fed the basic diet without chillies, the authors stress, that it cannot be said whether chillies have a specific carcinogenic effect or whether a deficiency in the diet aggravated by a non-specific irritant caused the tumours (Hoch-Ligeti, 1951). Capsaicin (capsaicin 65%, dihydro- 31%, nordihydro- 0.9%, homo- 1%, homodihydro- 0.6%, nor- 0.5%, nornor- 0.3%), administered in a semisynthetic diet at 0.03125% to 50 male and 50 female Swiss albino mice for their life span from 6 weeks of age, induced benign polypoid adenomas of the caecum in 22% of females (p
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