The Strategy for the Development of Botanical Products

1. The Strategy for the Development of Botanical Products Chieh-Fu Chen Chinese Traditional Medicine Research and Development Fund Scientific Pharmaceutical Elite Co. National Yang-Ming University Taipei, Taiwan

2. USA, 1906, Federal Food and Drug Act, Interstate transport of adulterated or misbranded foods and drugs. • Drug Regulation in USA

3. B‧Requirement and characteristics of modern medicine ‧Safety first (安全第一) ‧Evidence based efficacy (有確據之療效) ‧Quality consistency (品質ㄧ致) ‧Insights into mechanism of action (瞭解作 用機理) ‧Drugs interaction potential (與其他藥物之 相互作用)

4. C‧Results of Modern New Drug Development ( NDD ) a . Increase in working load b . Prolonged the time require c . Increase in cost d . Decline in the number of new drug approved e . Uneven share of global market f . Drug withdrawals g . Merge of drug companies

5. a . Increase in working load Data points generated by large drug company 1990, 200,000 1995, 5 to 6 million 2000, 50 million

6. b . Prolonged the time require Years from the initiation of the pre-clinical study to final FDA approval of a new drug 1963-1969： 8.1 1970-1979 : 11.6 1980-1989 : 14.1 1990-1999 : 14.2

7. c . Increase in cost Average capitalized costs for new drug R & D 1960s 54 million 1970s 138 million 1980s 318 million 1990s 802 million • s 900-1,700 million Inflation-adjusted rated ：11.8% 2003, New indication & formulation：120-150 million

8. d . Decline in the number of new drug approved • New chemical entities approved by FDA. In 1980’s were 60 per year

9. NCI approved by FDA

10. The approved of antibiotics 1983 ~ 1988 16 1988 ~ 1992 14 1992 ~ 1997 10 1997 ~ 2002 7

11. e . Uneven share of global market Global drug market in 2005 (600 billions)

13. Global Botanical Drugs Market： Billion 2003 Majory located in Europe > Asia > North America > Japan

14. USA, France, China Japan, Italy, India Germany, UK, Spain 9 major markets account for72%of total in 2002

15. Top 20 of herbal supplement sales in USA in 2001 • Echinacea 紫錐花 • Garic 大蒜 • Ginko biloba 銀杏 • Saw Palmetto 鋸櫚 • Ginseng 人參 • Grape seed 葡萄子 • Green tea 綠茶 • St John’s wort 金絲桃 • Bilbery 山桑子 • Aloe vera 蘆薈

16. 11. Granberry 大果蔓越桔 Vaccinium macrocarpon 12. Milk Thistle 奶薊 13. Ginger 薑 14. Olive leaf 橄欖葉 15. Mahuang 麻黃 16. Dong guai 當歸 17. Black cohosh 北美昇麻 18. Astragal 黃蓍 19. Siberian ginseng 20. Psyllium 洋車前 Plantago psyllium

17. f . Drug withdrawals US and European drug withdrawals for safety reasons (1982-1992) ＊ Drug name Safety issue Approximate months on the market United States and Europe Benoxaprofen Fatal cholestatic jaundice 4 Nomifensine Haemolytic anemia 7 Suprofen Flank pain syndrome 16 Ticrynafen Hepatotoxicity 8 Zomepirac Anaphyactoid reactions 29 Europe Cyanidanol Haemolytic anaemia ~6‡ Indoprophen Hepatotoxicity ~6‡ Isoxicam Epidemal necrolysis ~2‡ Sulocidil Hepatotoxicity ?‡ Zimelidine Hypersensitivity reactions with ?‡ Neurological involvement ＊US data from REF.30. ‡Never approved for sale in the United states.

18. Modified from REF.30. *Alosetron is now restricted to women with diarrhoea-predominant irritable bowel syndrome who have not responded to conventional therapies and who have been educated about the relativerisks and benefits 31. + Withdrawn from the market, but in some countries on restricted use.

19. § Discontinued the sale and distribution August 2003.

20. g . Merge of drug companies

21. Pfizer/ Warner-Lambert R & D in 1999 : 4 billionPharmacia & john/Monsanto R & D in 1999 : 2.8 billion

22. Merge of above two in 2003 to create the world’s largest pharmaceutical companyR & D expenditure is planned to be 7.9 billion in 2004(Total revenues : 54 billion Net income : 12.8 billion)

23. C . Old age Society & Drug Ｔhe percentage of population which over 65 in Taiwan

24. In China 143 million ( 11% ) of population are over 60 in 2006In 2020, it will be 17%In 2050, it will reach 31% (over 400 million)Thus, treatment of multiple factorial diseases such as hypertension, neurodegeneration, cancer etc are most impotant in now and future

25. Most chronic diseases and aging-related syndromes are caused by multifactors. Take neurodegeneration for example, the lipid peroxidation, DNA and protein damage and finally neuronal death are caused by selective vulnerability, genetics, environmental triggers, excitotoxicity, energy metabolism and aging, and oxidative stress.

26. Thus, it is impossible to treat then by single drug. On the other hand, the toxic effect of a drug is a function of dosage. The greater the amount of a chemical that must be used to produce the therapeutic effect, the greater is the chance of the toxic effect.

27. D‧Pharmaceuticals market in China Pharmaceutical sales in China are US$25.5 bn. in 2005 A 8.5% increase over the previous year Among them Traditional Chinese Medicines are estimated as US$5.8 bn.

28. Thus, the size of western-drug market is US$19.2 bn, equal to around US$15 per capita, comparable with India. However, this is one of the largest markets in the world, and largest in Asia behind Japan. In US dollars terms, China could overtake Germany in the next four years, Japan by 2015, and the US by 2039

29. E‧The question arose：can the poor people using good drug ? F‧Strategies to conquer such problem are Supply of essential drugs Using of generic drug Well using the herbal drug G‧What are the problem of herbal drugs

30. Thus , and herb contains not only one active constituent, and most bioactive constituents have several bioactivities. Furthermore, the content of bioactive constituents in herbs as affected by various factors, soil, quantity and quality of water, temperature, sunshine, harvest time, and processing, increases this complexity.

31. It is difficult right now to determine how many chemicals are needed and what their ratio should be to treat the multiple factorial chronic or aging related diseases.

32. H‧Chemical & biological fingerprints are dual-seal of botanical products quality Quality control is not only the guarantee for continuous development of TCM or botanical product, but also the premier for its globalization

33. Chemical fingerprints Besides taxonomy, morphology, histology, and analytical chemistry, the chemical fingerprint spectrum of TCM is considered as an important supplement to quantifying the marker component as assess by HPLC, IR, NMR, LC/MS/MS.

35. 漢防己質譜層析圖 漢防己質譜UV層析圖

36. 漢防己活性成分質譜圖

37. Biological fingerprints (1)‧Using enzymes receptor, cells, tissues, isolated organ assays to establish in vitro biological fingerprints (2)‧ Observing the drug-cytochromes interaction (CYP induction or inhibition) (3)‧ To perform in vivo studies to elucidate or confirm the in vitro findings

38. I‧In the past few years several of our efforts related to the herbal drugs are reported. They are :

39. 1、Chemical fingerprint : • Determination of tetrandrine, fangchinoline, cyclanoline and oblongine in Radix Stephaniae tetrandrae by high-performance liquid chromatography, J. Chin. Med., 13(1)39-48. (2002) • Determination of dehydroevodiamine, evodiamine, rutaecarpine and synephrine in Evodia genus plants from Taiwan and mainland China, J.Chin. Med., 13(3):151-158. (2002) • Quantitatively evaluation of bioactive components of Evodia rutaecarpa (Tetradium ruticarpum) in different harvesting times, Chin. Pharm. J., 55:305-312. (2003)

40. 2、Drug drug interaction : • The neuroprotective effects of phytoestrogens on amyloid β protein-induced toxicity are mediated by abrogating the activation of caspase cascade in rat cortical neurons, J. Biol. Chem., 276(7):5287-5295. (2001) • Herb-drug interaction of Evodia rutaecarpa extract on the pharmacokinetics of theophylline in rats, J. Ethnopharmacol ,102:440-445. (2005)

41. 3、Drug and CYP interaction : • Induction of cytochrome P450-dependent monooxygenase in mouse liver and kidney by rutaecarpine, an alkaloid of the herbal drug Evodia rutaecarpa. Life Sci., 70(2): 207-217. (2001) • Effects of CVT-E002, a proprietary extract from the north american ginseng (Panax quinquefolium) on hepatic drug-metabolizing enzymes in C57Bl/6J mice, J. Chin. Med., 13(2):89-96. (2002) • Effects of Wu-chu-yu- tang and its component herbs on drug-metabolizing enzymes, Jpn. J. Pharmacol., 89:267-273. (2002) • The alkaloid rutaecarpine is a selective inhibitor of cytochrome P450 1A in mouse and human liver microsomes, Drug Metab. Dispos., 30:349-353. (2002) • Modulation of drug-metabolizing enzymes by extracts of a herbal medicine Evodia rutaecarpa in C57BL/6J mice, Life Sci., 71:1267-1277. (2002) • Induction of CYP1A by a diterpene quinone tanshinone IIA isolated from a medicinal herb Salvia miltiorrhiza in C57BL/6J but not in DBA/2J mice, Life Sci., 74(7):885-896. (2004) • Identification of the microsomal oxidation metabolites of rutaecarpine, a main active alkaloid of medicinal herb Evodia rutaecarpa, J. Chromatogr A, 1076:103-109. (2005) • Characterization of mouse cytochrome P450-catalyzed oxidative metabolism of rutaecarpine, an alkaloid of the herbal medicine Evodia rutaecarpa, J. Food Drug Anal., (in press). (2006) • Oxidative metabolism of the alkaloid rutaecarpine by human cytochrome P450s, Drug Metab Dispos, 34:821-827. (2006)

42. 4、Mechanism : • Vasorelaxing action of rutaecarpine: effects of rutaecarpine on calcium channel activities in vascular endothelial and smooth muscle cells, J. Pharmacol. Exp. Ther., 289(3):1237-1244.(1999) • Andrographolide acts through inhibition of ERK1/2 and Akt phosphorylation to suppress chemotactic migration. Eur. J. Pharmacol., 498: 45-52. (2004) • Calcium-antagonizing activity of S-petasin, a hypotensive sesquiterpene from Petasites formosanus, on inotropic and chronotropic responses in isolated rat atria and cardiac myocytes, Naunyn. Schmiedebergs. Arch. Pharmacol., 369(3):322-329. (2004) • Prevention of macrophage adhesion molecule-1 (Mac-1)- dependent neutrophil firm adhesion by taxifolin through impairment of protein kinase- dependent NADPH oxidase activation and antagonism of G protein-mediated calcium influx, Biochem. Pharmacol., 67:2251-2262. (2004) • Tournefolic acid B methyl ester attenuates glutamate-induced toxicity by blockade of ROS accumulation and abrogating the activation of caspases and JNK in rat cortical neurons, J. Neurochem., 92:692-700. (2005) • Lipopolysaccharide-stimulated responses in rat aortic endothelial cells by a systems biology approach. Proteomics, (In press).(2006) • Taxifolin ameliorates cerebral ischemia-reperfusion injury in rats through its anti-oxidative effect and modulation of NF-kappa B activation. J. Biomed. Sci. 13(1):127-141. (2006)

43. 5、Partial purified herbal preparations : • Anti-Inflammatory Effects of the Partially Purified Extract of Radix Stephaniae tetrandrae: Comparative Studies of Its Active PrinciplesTetrandrine and Fangchinoline on Human Polymorphonuclear Leukocyte Functions, Mol. Pharmacol., 60(5):1083-1090. (2001) • Cardiac effects of the extract and active components of Radix stephaniae tetrandraeI. Electrically-induced intracellular calcium transient and protein release during the calcium paradox, Life Sci., 68(25):2853-2861. (2001) • Cardiac effects of the extract and active components of Radix stephaniae tetrandrae II. Myocardial infarct, arrhythmias, xoronary arterial flow and heart rate in the isolated perfused rat heart, Life Sci., 68(25):2863-2872. (2001) • Special processed Panax ginseng (SPPG) relaxes isolated rabbit corpus cavernosum through histamine antagonized property and with a beneficial effect in raising intracavernous pressure, J. Chin. Med., 13(4):197-208. (2002) • Evodia rutaecarpa protects against circulation failure and organ dysfunction in endotoxaemic rats through modulating nitric oxide release, J. Pharma. Pharmacol., 54:1399-1405. (2002) • Antihypertensive and anti-arrhythmic effects of an extract of Radix Stephaniae tetrandrae in the rat, J.Pharm. Pharmacol., 67:2251-2262. (2004)

44. 6、Pharmackinetics : • Measurement and pharmacokinetic of unbound 20(S)-camptothecin in rat blood and brain by microdialysis coupled to microbore liquid chromatography with fluorescence detection, J. Chromatogr. A, 870(1-2):221-226. (2000) • Determination of chlorogenic acid in rat blood by microdialysis coupled with microbore liquid chromatography and its application to pharmacokinetic studies, J. Chromatogr. A, 870(1-2):443-448. (2000) • The effects of the cyclosporin A, a P-glycoprotein inhibitor, on the pharmacolinetics of baicalein in the rat：a microdialysis study, Br. J. Pharmacol., 137:1314-1320. (2002) • Identification of the microsomal oxidation metabolites of rutaecarpine, a main active alkaloid of medicinal herb Evodia rutaecarpa, J. Chromatogr A, 1076:103-109. (2005) • Elimination of rutaecarpine and its metabolites in rat feces and urine measured by liquid chromatography, Biomed Chromatogr. (in press). (2006)

45. J‧Conclusion： • The develop of so-called new chemical entity drug is very expensive, and more than 50% of the people are too poor to use such new drugs. 2. The cost for the development of botanical products are more reasonable. 3. The most potential botanical products markets are China, India, and other developing or under-developing areas.

46. 4. Devolop of well chemical and biological controlled partial purified herbal drug or its compounding preparation to new drug is one of the best way in the utilize the herbal medicine, both in cost and time saving