RESEARCH THE CLINIC OF THE COLLEGE OF CHINESE MEDICINE HERBAL MEDICINE FOR CANCER PATIENTS Kirsten Dhar – The Clinic of the College of Chinese Medicine, 26-28 Finchley Rd, St John’s Wood, London, NW8 6ES Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development ABSTRACTS – TRIALS – RESEARCH ARTICLES Introduction to plant synergy Abstracts from the journal ‘Molecular Cancer Therapeutics’ (American Association for Cancer Research) Summaries of Abstracts and Trials Herbs for Cancer Introduction to plant synergy In Traditional Chinese Medicine (TCM), herbal prescriptions are given to patients as complex formulations containing multiple herbs. Notably, this approach amounts to the administration of several chemical entities at once. The underlying theory is, that interaction among the chemicals present within a single plant, as an entity, and also the different herbs in a formula exert synergistic pharmaco-dynamic actions not present when administered in form of an isolated active ingredient. Synergy of many different chemical components within the plant is responsible for these effects and involves a number of different processes. Whilst the exact mechanisms behind plant synergy and the allelopathic properties of the plant are still not fully understood, a great deal of research has been done in this field. Today’s understanding of basic molecular cell biochemistry and the consequent possibility of elucidating the actual mechanism of action of chemical components at the cellular level, or even gene level, have resulted in a rational explanation of the action of medicinal plants. In therapeutic use, medicine administered in form of the entire plant often shows higher plasma availability of the active ingredient than when given in an isolated and concentrated form. Since active Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development components within the plant are rather low in concentration, it has become evident that there is an involvement of substances which are inactive medicinally but enhance the activity of and protect the integrity of the so-called active principles of the plant. In the case of Artemisia annua, a plant which is used to treat malaria, the isolated active principle artemisinine (2), administered orally at 3000mg over 4 days produced the same result (anti-parasitic, anti-pyretic, recrudescence rate) as a four day course of the tea leaves containing a total dose of 50mg artemisinine. Plant synergy also offers protection of an active ingredient from degradation by enzymes and it facilitates transport across barriers such as cell and organelle membranes. Passage of active ingredients through membranes, which occurs in the gut from the intestine wall into the blood stream through to the penetration of an infected cell or into a parasite, is another important aspect of synergistic properties. There is evidence of protective action and active transport across, for example, intestinal walls and inhibition or reversal of the excretory process which naturally takes place in this location. One of the key componants to such improved transport of a substance through the cell membrane are flavonoids. Plant tissue contains phospholipids, which may be partially or entirely lost in the extraction process when isolating the active ingredient. These phospholipids, however, are crucial in transport as they interact between polar heads of the phospholipids molecules of the membrane and the phenolic hydroxyl groups of flavonoids in the process of phospholipic bilayer diffusion. Moreover, plant synergy can neutralize the adverse effects and toxicities of specific individual chemicals and provide other signals to the host’s cells that result in higher efficacy of the crude drug when compared with isolated components. It can also overcome and, sometimes reverse, multi-drug resistance mechanisms in patients who are on medication over prolonged periods of time and in chemotherapy. MDR protein inhibitors are present in the plant and are biologically active when the whole extract is used rather than an isolated principle. There is no doubt that many other molecular mechanisms are involved in the behaviour often observed in chemical activity when the ingredient is retained in the original crude extract and not all of them are, as yet, fully understood. Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development Abstracts from Molecular Cancer Therapeutics Reversal of cisplatin resistance with a BH3 mimetic, (–)gossypol (Chinese Cotton Seed Plant), in head and neck cancer cells: role of wild-type p53 and Bcl-xL Mol Cancer Ther. 2005;4:1096-1104 Joshua A. Bauer1, Douglas K. Trask2, Bhavna Kumar2, Gerrit Los6, Jason Castro2, Julia ShinJung Lee4,5, Jianyong Chen3, Shaomeng Wang3,5, Carol R. Bradford2,5 and Thomas E. Carey1,2,5 Departments of 1 Pharmacology, 2 Otolaryngology-Head and Neck Surgery, and 3 Internal Medicine and Medicinal Chemistry; 4 Biostatistics Unit; and 5 University of Michigan Comprehensive Cancer Center, University of Michigan, Ann Arbor, Michigan; and 6 Pfizer Global Research and Development, La Jolla, California Requests for reprints: Thomas E. Carey, Department of Otolaryngology, University of Michigan, 6020 KHRI, 1301 East Ann Street, Ann Arbor, MI 48109-0506. Phone: 734-764-4371; Fax: 734-764-0014. Background: Multi-drug resistance (MDR) is an important biological behaviour of tumour cells in chemotherapy and it is also one of the major causes of clinical chemotherapy failure. There is increasing interest into the exact mechanism of tumour cells' MDR and its reversion by Chinese herbs. Organ preservation protocols in head and neck squamous cell carcinoma (HNSCC) are limited by tumours that fail to respond. We observed that larynx preservation and response to chemotherapy is significantly associated with p53 overexpression, and that most HNSCC cell lines with mutant p53 are more sensitive to cisplatin than those with wild-type p53. To investigate cisplatin resistance, we studied two HNSCC cell lines, UM-SCC-5 and UM-SCC-10B, and two resistant sublines developed by cultivation in gradually increasing concentrations of cisplatin. The cisplatin-selected cell lines, UM-SCC-5PT and UM-SCC-10BPT, are 8 and 1.5 times more resistant to cisplatin than the respective parental cell lines, respectively. The parental lines over-express p53 and contain p53 mutations but the cisplatin-resistant cell lines do not, indicating that cells containing mutant p53 were eliminated during selection. Bcl-xL expression increased in the cisplatinresistant lines relative to the parental lines, whereas Bcl-2 expression was high in the parental lines and decreased in the cisplatin-resistant lines. Thus, cisplatin selected for wild-type p53 and high Bcl-xL expression in these cells. We tested a smallmolecule BH3 mimetic, (–)-gossypol, which binds to the BH3 domain of Bcl-2 and Bcl-xL, for activity against the parental and cisplatin-resistant cell lines. At physiologically attainable levels, (–)gossypol induces apoptosis in 70% to 80% of the cisplatin-resistant cells but only in 25% to 40% of the parental cells. Thus, cisplatinresistant cells seem to depend on wild-type p53 and Bcl-xL for survival and BH3 mimetic agents, such as (–)-gossypol, may be useful adjuncts to overcome cisplatin resistance in HNSCC. Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development Triptolide Inhibits the Growth and Metastasis of Solid Tumours Vol. 2, 65-72, January 2003, Molecular Cancer Therapeutics Shanmin Yang, Jinguo Chen, Zhen Guo, Xue-Ming Xu, Luping Wang, Xu-Fang Pei, Jing Yang, Charles B. Underhill and Lurong Zhang2 Department of Oncology, Georgetown University Medical Center, Washington, DC 20007 [S. Y., J. C., X-M. X., L. W., X-F. P., J. Y., C. B. U., L. Z.], and Key Laboratory of China Education Ministry on Cell Biology and Tumour Cell Engineering, Xiamen University, Fujian, People’s Republic of China 361003 [S. Y., Z. G., L. Z.] Triptolide (TPL), a diterpenoid triepoxide purified from the Chinese herb Tripterygium wilfordii (Lei Gong Teng) was tested for its antitumour properties in several model systems. In vitro, TPL inhibited the proliferation and colony formation of tumour cells at extremely low concentrations (2–10 ng/ml) and was more potent than Taxol. Likewise, in vivo, treatment of mice with TPL for 2–3 weeks inhibited the growth of xenografts formed by four different tumour cell lines (B16 melanoma, MDA-435 breast cancer, TSU bladder cancer, and MGC80-3 gastric carcinoma), indicating that TPL has a broad spectrum of activity against tumours that contain both wildtype and mutant forms of p53. In addition, TPL inhibited experimental metastasis of B16F10 cells to the lungs and spleens of mice. The anti-tumour effect of TPL was comparable or superior with that of conventional anti-tumour drugs, such as Adriamycin, mitomycin, and cisplatin. Importantly, tumour cells that were resistant to Taxol attributable to the over-expression of the multidrug resistant gene 1 were still sensitive to the effects of TPL. Studies on cultured tumour cells revealed that TPL induced apoptosis and reduced the expression of several molecules that regulate the cell cycle. Taken together, these results suggest that TPL has several attractive features as a new anti-tumour agent. PG490-88, a derivative of triptolide (an active ingredient of some Chinese medicinal herbs), causes tumor regression and sensitizes tumors to chemotherapy Mol Cancer Ther. 2003;2:855-862 John M. Fidler1, Ke Li3, Cathie Chung2, Ke Wei2, Jessica A. Ross2, Mingxing Gao2 and Glenn D. 2 Rosen2 Pharmagenesis, Inc., Palo Alto, CA; Department of Medicine, Stanford 3 University Medical School, Stanford, CA; and Applied Biosystems, Foster City, CA Treatment of solid tumours with combinations of chemotherapeutic agents has not led to significant increases in long-term survival. Recent studies support a role for inhibitors of checkpoint arrest as a means to enhance the cytotoxicity of chemotherapy. We have shown previously that triptolide (PG490), an oxygenated diterpene derived from a Chinese medicinal plant, induces apoptosis in Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development cultured tumour cells and sensitizes tumor cells to topoisomerase inhibitors by blocking p53-mediated induction of p21. Here we extend our studies to a tumour xenograft model and evaluate the efficacy and safety of PG490-88 (14-succinyl triptolide sodium salt), a water-soluble prodrug of PG490. We also look at the combination of PG490 or PG490-88 with CPT-11, a topoisomerase I inhibitor, in cultured cells and in the tumour xenograft model. We show that PG490-88 is a safe and potent anti-tumour agent when used alone, causing tumour regression of lung and colon tumour xenografts. We also show that PG490-88 acts in synergy with CPT-11 to cause tumour regression. A phase I trial of PG490-88 for solid tumours began recently and safety and optimal dosing data should accrue within the next 12 months. Our findings that PG490-88 causes tumour regression and that it acts in synergy with DNA-damaging chemotherapeutic agents suggest a role as an antineoplastic agent and chemosensitizer for the treatment of patients with solid tumours. Caspase-dependent and caspase-independent apoptosis induced by evodiamine in human leukemic U937 cells Mol Cancer Ther. 2006;5:2398-2407 Tae-Jin Lee1, Eun Jung Kim1, Shin Kim1, Eun Mi Jung1, Jong-Wook Park1, Seung Hun Jeong2, Sang Eun Park2, Young Hyun Yoo2 and Taeg Kyu Kwon1 1 Department of Immunology and Chronic Disease Research Center and Institute for Medical Science, School of Medicine, Keimyung University, Taegu, South Korea and 2 Department of Anatomy and Cell Biology, Dong-A University College of Medicine (BK21 Program), and Medical Science Research Center, Busan, South Korea Evodiamine is one of the major bioactive compounds that have been isolated and purified from the fruit of Evodiae fructus (Wu Zhu Yu). Evodiamine exhibits anti-tumour activities against the human tumor cells, including multi-drug-resistant tumour cells. However, the molecular mechanism involved in cell death induced by evodiamine treatment remains poorly understood. In the present study, we showed that evodiamine activated the caspase-dependent apoptotic pathway. This apoptosis was only partially inhibited by a pancaspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone, which suggested that evodiamine-induced apoptosis in leukemic U937 cells is partially caspase independent. We observed the nuclear translocation of apoptosis-inducing factor in evodiamineinduced apoptosis of U937 cells, which may be responsible for the caspase-independent apoptotic execution. We next showed that evodiamine induced the substantial amount of apoptosis both in Bcl2- and Akt-overexpressing U937 cells but not in human peripheral blood mononuclear cells. Although benzyloxycarbonyl-Val-Ala-Aspfluoromethyl ketone inhibited caspase activity in Bcl-2overexpressing U937 cells, it completely prevented neither the induction of apoptosis or the nuclear translocation of apoptosisKirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development inducing factor, which suggests that evodiamine is, at least in part, able to bypass the resistance of leukemia cells via caspaseindependent apoptotic pathways. Thus, therapeutic strategy using evodiamine may warrant further evaluation. Boswellic acid acetate induces apoptosis through caspasemediated pathways in myeloid leukaemia cells Mol Cancer Ther. 2005;4:381-388 1 Lijuan Xia1, Duo Chen1, Rui Han2, Qicheng Fang2, Samuel Waxman1 and Yongkui Jing1 Division of Hematology/Oncology, Department of Medicine, Mount Sinai School of Medicine, New York, New York and 2 Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing, China The Chinese herb Ru Xiang (Boswellia carterii) is investigated for its antineoplastic properties in this article. The mechanism of the cytotoxic effect of boswellic acid acetate, a 1:1 mixture of boswellic acid acetate and ß-boswellic acid acetate, isolated from Boswellia carterri Birdw on myeloid leukaemia cells was investigated in six human myeloid leukaemia cell lines (NB4, SKNO-1, K562, U937, ML-1, and HL-60 cells). Morphologic and DNA fragmentation assays indicated that the cytotoxic effect of boswellic acid acetate was mediated by induction of apoptosis. More than 50% of the cells underwent apoptosis after treatment with 20 µg/mL boswellic acid for 24 hours. This apoptotic process was p53 independent. The levels of apoptosis-related proteins Bcl-2, Bax, and Bcl-XL were not modulated by boswellic acid acetate. Boswellic acid acetate induced Bid cleavage and decreased mitochondrial membrane potential without production of hydrogen peroxide. A general caspase inhibitor (Z-VAD-FMK) and a specific caspase-8 inhibitor II (Z-IETDFMK) blocked boswellic acid acetate–induced apoptosis. The mRNAs of death receptors 4 and 5 (DR4 and DR5) were induced in leukaemia cells undergoing apoptosis after boswellic acid acetate treatment. These data taken together suggest that boswellic acid acetate induces myeloid leukaemia cell apoptosis through activation of caspase-8 by induced expression of DR4 and DR5, and that the activated caspase-8 either directly activates caspase-3 by cleavage or indirectly by cleaving Bid, which in turn decreases mitochondria membrane potential. Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development Effect of curcumin on normal and tumour cells: Role of glutathione and bcl-2 Mol Cancer Ther. 2004;3:1101-1108 Christine Syng-ai, A. Leela Kumari and Ashok Khar - Center for Cellular and Molecular Biology, Hyderabad, India Curcumin, a well-known dietary pigment derived from Curcuma longa, inhibited growth of several types of malignant cells both in vivo and in vitro. However, its mechanism of action still remains unclear. In this study, we have focused primarily on the cytotoxic effects of curcumin on three human tumour cell lines and rat primary hepatocytes. Curcumin induced apoptosis in MCF-7, MDAMB, and HepG2 cells in a dose-dependent and time-dependent manner. Apoptosis was mediated through the generation of reactive oxygen species. Attempts were made to establish the role played by endogenous glutathione on the apoptotic activity of curcumin. Depletion of glutathione by buthionine sulfoximine resulted in the increased generation of reactive oxygen species, thereby further sensitizing the cells to curcumin. Interestingly, curcumin had no effect on normal rat hepatocytes, which showed no superoxide generation and therefore no cell death. These observations suggest that curcumin, a molecule with varied actions, could be developed into an effective chemopreventive and chemotherapeutic agent. Berberine (Huang Lian, Rhizoma Coptidis) , a natural product, induces G1-phase cell cycle arrest and caspase-3dependent apoptosis in human prostate carcinoma cells Sudheer K. Mantena1, Som D. Sharma1 and Santosh K. Katiyar1,2,3,4 Departments of 1 Dermatology, 2 Environmental Health Sciences, 3 Clinical Nutrition Research Center, and 4 Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama Berberine, a naturally occurring isoquinoline alkaloid, has been shown to possess anti-inflammatory and anti-tumour properties in some in vitro systems. Here, we report that in vitro treatment of androgen-insensitive (DU145 and PC-3) and androgen-sensitive (LNCaP) prostate cancer cells with berberine inhibited cell proliferation and induced cell death in a dose-dependent (10–100 µmol/L) and time-dependent (24–72 hours) manner. Treatment of non-neoplastic human prostate epithelial cells (PWR-1E) with berberine under identical conditions did not significantly affect their viability. The berberine-induced inhibition of proliferation of DU145, PC-3, and LNCaP cells was associated with G1-phase arrest, which in DU145 cells was associated with inhibition of expression of cyclins D1, D2, and E and cyclin-dependent kinase (Cdk) 2, Cdk4, and Cdk6 proteins, increased expression of the Cdk inhibitory proteins (Cip1/p21 and Kip1/p27), and enhanced binding of Cdk inhibitors to Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development Cdk. Berberine also significantly (P < 0.05–0.001) enhanced apoptosis of DU145 and LNCaP cells with induction of a higher ratio of Bax/Bcl-2 proteins, disruption of mitochondrial membrane potential, and activation of caspase-9, caspase-3, and poly(ADPribose) polymerase. Pre-treatment with the pan-caspase inhibitor zVAD-fmk partially, but significantly, blocked the berberine-induced apoptosis, as also confirmed by the comet assay analysis of DNA fragmentation, suggesting that berberine-induced apoptosis of human prostate cancer cells is mediated primarily through the caspase-dependent pathway. The effectiveness of berberine in checking the growth of androgen-insensitive, as well as androgensensitive, prostate cancer cells without affecting the growth of normal prostate epithelial cells indicates that it may be a promising candidate for prostate cancer therapy. SUMMARIES OF ABSTRACTS AND TRIALS Ginseng enhances anti-cancer effect of 5-fluorouracil on human colorectal cancer cells Wang CZ, Luo X, Zhang B, Song WX, Ni M, Mehendale S; Tang Center for Herbal Medicine Research, The University of Chicago; Cancer Chemotherapy and Pharmacology Journal; 2006 Sep 29; PMID: 17009031 [PubMed - as supplied by publisher] PURPOSE: Panax notoginseng is a commonly used Chinese herb. Although a few studies have found that notoginseng shows antitumor effects, the effect of this herb on colorectal cancer cells has not been investigated. 5-Fluorouracil (5-FU) is a chemotherapeutic agent for the treatment of colorectal cancer that interferes with the growth of cancer cells. CONCLUSIONS: This study demonstrates that the herb can enhance the anti-proliferation effect of 5-FU on HCT-116 human colorectal cancer cells and may decrease the dosage of 5-FU needed for colorectal cancer treatment. Wogonin sensitizes resistant malignant cells to TNF-alphaand TRAIL-induced apoptosis Fas SC, Baumann S, Zhu JY, Giaisi M, Treiber MK, Mahlknecht U, Krammer PH, Li-Weber M; Tumor Immunology Program D030, German Cancer Research Center (DKFZ), Published: Blood, 2006 Dec 1; 1;108(12):3700-6; PMID: 16931628 [PubMed - indexed for MEDLINE] We show here that wogonin, derived from the popular Chinese herb Huang-Qin, attenuates NF-kappaB activity by shifting TNFalphaKirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development induced free radical .O(2)(-) to a more reduced nonradical product, H(2)O(2), and thereby sensitizes TNFalpha-resistant leukemia cells to TNFalpha-induced apoptosis. Importantly, wogonin does not affect the viability of normal peripheral blood T cells. Wogonin also sensitizes TRAIL-induced apoptosis. Our data suggest a potential use of wogonin as a TNFalpha or TRAIL adjuvant for cancer treatment. Our data also demonstrate how a herbal compound enhances killing of tumor cells with reduced side effects compared with other treatments. Potent inhibition of Lewis lung cancer growth by heyneanol A from the roots of Vitis amurensis through apoptoic and antiangiogenic activities Lee EO, Lee HJ, Hwang HS, Ahn KS - Graduate School of East-West Medical Science, Kyunghee University, Kiheungeup, Yongin 449-701, Republic of Korea. Published: Carcinogenesis, 2006 Oct;27 (10):2059-69; PMID: 16675471 [PubMed - indexed for MEDLINE] Vitis amurensis Rupr. (Shan Pu Tao) has long been used for the treatment of cancer to this end, we isolated from its root heyneanol A (HA), which is a tetramer of resveratrol (RES), and established the in vivo anti-tumour activity of HA using the Lewis lung carcinoma (LLC) model. Remarkably, HA was fairly stable in cell culture medium and did not undergo intracellular conversion to RES. Therefore, HA is an active anti-cancer compound that induces caspase-mediated cancer cell apoptosis and inhibits angiogenesis rivaling the potency of RES and merits further evaluation for cancer chemoprevention. Effect of lyceum barbarum polysaccharide on human hepatoma QGY7703 cells: inhibition of proliferation and induction of apoptosis Zhang M, Chen H, Huang J, Li Z, Zhu C, Zhang S - Department of Food Science and Bioengineering, Tianjin University of Science and Technology, Tianjin; PMID: 15826878 [PubMed - indexed for MEDLINE] Lycium barbarum polysaccharide (LBP), extracted from Lycium barbarum (Fructus Lycii – Gou Qi Zi and Cortex Lycii – Di Gu Pi) is found to have anticancer activity. In this study, the effect of LBP on the proliferation rate, cell cycle distribution and apoptosis in the human hepatoma QGY7703 cell line were investigated. CONCLUSION: The induction of cell cycle arrest and the increase of intracellular calcium in apoptotic system may participate in the antiproliferative activity of LBP in QGY7703 cells. Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development Differential control of growth, cell cycle progression and gene expression in human estrogen receptor positive MCF-7 breast cancer cell by extracts derived form polysacchaaropeptide I’m-Yunity (a mushroom extract) and Dan Shen (Radix Salviae Miltiorrhizae) Hsieh TC, Wu JM - Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla; Published: International Journal of Oncology, 2006 Nov;29(5):1215-22; PMID: 17016654 [PubMed - indexed for MEDLINE] In this study, we investigated the effects of 70% ethanolic extracts prepared from medicinal mushroom extract denoted I'm-Yunity and Dan Shen, alone and in combination, using MCF-7 cells as an in vitro model of estrogen receptor positive (ER+), low invasive Breast Cancer. CONCLUSION: Treatment by I'm-Yunity did not affect cell cycle progression in MCF-7 cells; however, it promoted active induction of apoptosis. In addition, treatment with Danshen alone resulted in a pronounced reduction in the expression of Rb, cyclin D1, and p53, and also led to a diminution of p65 and p50 forms of NF-kappaB. The pronounced suppressive effects of Danshen on expression of the aforementioned genes were largely attenuated in cells treated with I'm-Yunity-Plus suggesting that ingredients in Danshen must have interacted with those in I'm-Yunity as to culminate in neutralization of the gene suppressive effects of Danshen. Additional support for such interactions was obtained by targeted cDNA array analysis using human tumor metastasis and BCa/ER signaling gene arrays. Taken together, our results are consistent with the interpretation that interaction exists between Danshen and I'm-Yunity and that I'm-Yunity-Plus may have very good efficacy in the treatment of Breast Cancer, particularly for patients with ER+ status. Evidence for Oldenlandia diffusa-evoked cancer cell apoptosis through superoxide burst and caspase activation Yadav SK, Lee SC - Yong Loo Lin School of Medicine, National University of Singapore; Zhong Xi Yi Jie He Xue Bao, 2006 Sep; 4(5):485-9; PMID: 16965742 [PubMed - in process] BACKGROUND & OBJECTIVE: Various studies using the herb Oldenlandia diffusa (Bai Hua She She Cao) alone or in combination with other therapy plans have evidenced the effectiveness of the herb in the management of cancers of different tissue origin. RESULTS: We found that it dose-dependently inhibited the cancer cell growth in MTT assay. Flow cytometry analysis revealed that it elicited significant production of sub-G(1) population of the cells, indicating the extract-evoked cell apoptotic death. The LD(50) of the ethanol extract was estimated to be approximately 320 microg/ml. Moreover, treatment of the cancer cells with the ethanol component markedly increased the production of superoxide within few hours. Significant elevation in the protease activities of Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development caspases-2 and -3 were detected at as early as 3 and 6 hours respectively. CONCLUSION: Our results show that the ethanol extract of the herb effectively evokes cancer cell apoptosis, possibly through burst-mediated caspase activation. Chinese Herb/Chemotherapy Interactions in Breast Cancer University of California, San Francisco, Michael Campbell Ph D, Award: 2003 (Cycle IX) $200,000 #9WB-0042 Innovative Treatment Modalities: Hormone and Chemotherapy Targets – Improving Today’s Arsenal. Traditional Chinese Medicine (TCM) is a treatment modality that has been used for centuries to treat cancer, prolong life, and increase the quality of life for cancer patients. The lack of evidence-based clinical trials coupled with the proliferation of anecdotal and case report studies of TCM make it an important treatment modality to study. TCM use in the San Francisco Bay Area is widespread and frequently used by our patient population, thus it can be studied using standard research models at our institution. There is a high degree of conformity among TCM practitioners in terms of diagnoses and treatments due to the extensive historically-based documentation of TCM treatment modalities. We have recently obtained laboratory data on the effects of over 70 Chinese herbal extracts on the growth of breast cancer cells in culture. These herbs have been traditionally prescribed by TCM practitioners for metastatic breast cancer and other cancer patients. Many of these herbal extracts demonstrated significant inhibitory effects in our initial screening. Since many cancer patients who take these herbs also receive conventional chemotherapy, the objective of this project is to determine what kinds of interactions these herbs have with standard chemotherapeutic agents. Herbs will be prepared as boiled teas, as they are traditionally prepared. Breast cancer cells growing in tissue culture will be treated with individual herbs, chemotherapeutic drugs, or combinations of an herb with a chemotherapeutic drug. After three days of treatment, the cells will be analyzed for growth/death. Growth curves will be generated from these data and comparisons of the curves from individual herbs, drugs, and their combinations will determine whether the herbs act to enhance the effects of the drugs, inhibit their effects, or have no effect. We will then examine the effects of herb/drug combinations that show greater activity than either alone in a mouse model of breast cancer. Rather than randomly screening thousands of plants for anti-breast cancer activity, we have focused on plants which have been used for hundreds/thousands of years in Chinese medicine. In addition, we are administering these agents as “teas” which is how they are traditionally administered to patients. Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development Laboratory Testing of Chinese Herbs Used for Breast Cancer University of California, San Francisco – Dabasish Tripathy MD, Award: 1999 (Cycle V) $85,754 Innovative Award: Creative Drugs. Traditional Chinese Medicine (TCM) is a treatment modality that has been used for centuries to cure cancer, prolong life, increase the quality of life for cancer patients, and more recently to ameliorate the side effects of Western therapies. The lack of evidence-based clinical trials coupled with the proliferation of anecdotal and case report studies of TCM make it an important treatment modality for further study. TCM use in the Bay Area is widespread and frequently used by our patient population, thus it can be studied using standard research models at our institution. There is a high degree of conformity among TCM practitioners in terms of both diagnoses and treatments due to the extensive historically-based documentation of TCM treatment modalities. We propose to obtain laboratory data on herbs commonly used for breast cancer in order to prioritize herbal formulae for future Phase I/II clinical trials. We plan to perform laboratory analysis of a number of botanical agents that have been traditionally prescribed by TCM practitioners for metastatic breast cancer (MBC) patients. We have pilot data on a few herbal compounds that show anti-tumour activity, and we will use a similar expanded methodology to screen approximately 50 more botanical agents. We will test for the ability of these compounds to cause breast cancer cells to divide using an MTT assay, to commit suicide (apoptosis) using APO-BRDU and to grow using clonogenic assays. The compounds will be tested for their effects using both pulsed and continuous exposure. For herbal extracts with activity, the responsible components will be identified using salt exchange column and high performance liquid chromatography fractions. Promising herbs or combinations could then be entered into Phase I/II clinical trials using tumor response endpoints. The Use of Ginger and Turmeric in Cancer Treatments Vimala, S., et al. Anti-tumour promoter activity in Malaysian ginger rhizobia used in traditional medicine. British Journal of Cancer, Vol. 80, No. 1/2, April 1999, pp. 110-16 Ginger, turmeric and other members of the Zingiberaceae family of rhizomes have a long history of use in Malaysian traditional medicine. Ginger, for example, is widely used in the treatment of stomach problems, nausea, vomiting, epilepsy, sore throat, cough, bruises, wounds, childbirth, sore eyes, liver complaints, rheumatism, asthma, and many other disorders. Researchers at the Forest Research Institute of Malaysia now report that several members of the Zingiberaceae family effectively block the promotion of cancerous tumors. They tested 11 different species and found that seven of them had strong anti-tumor properties. Their test involved a short term assay of the inhibitory effect of extracts of the rhizomes (roots) on human cancer cells. They found that turmeric (Curcuma domestica) extracts (turmeric root extracted with petroleum ether, chloroform or ethanol) completely inhibited further growth of the cancer cells. Ginger (Zingiber officinale) extracts, especially the Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development chloroform extract, also inhibited further growth, but the concentration of extract was more critical than for the turmeric extracts. The researchers conclude that turmeric, ginger and other Zingiberaceae rhizomes may be useful in preventing the promotion of cancer and that populations with high risks of cancer should be encouraged to include them in their diet. Further work is now underway to isolate the active components in the plants. 3. HERBS FOR CANCER While it may not quite be a case of: “Eye of newt and toe of frog, Wool of bat and tongue of dog, Of the ravin’d salt-sea shark, Root of hemlock digg’d i’ the dark, Double, double, toil and trouble: Fire burn and cauldron bubble.” (William Shakespeare ‘Macbeth’ 1606)… …bioactive compounds from plants have been harvested and deployed for a variety of purposes, not least in health and medicine, since before the dawn of agriculture more than 10 000 years ago. The fact that many of the most potent pharmaceuticals and poisons known to man are of natural, plant origin is often over-looked by the more ardent proponents of all things ‘organic’! By the nineteenth century serious attempts were being made to provide a scientific rationale for observations of allelopathic phenomena. One of the best known identities of this period was Augustin Pyrame de Candolle. As early as 1805 he wrote on the topic of root excretions and applied these studies in the context of crop rotation. The technology of the day, however, did not permit him to identify the bioactive compounds which he perceived to be present. As Willis (2002) writes: “To many, de Candolle is the real father of allelopathy….he developed a perspicacious theory of plant interaction, relevant to agriculture and natural ecosystems, based on chemical substances assumedly released from plants”. Perhaps de Candolle should be recognized as the grand-father of allelopathy? In any event, it was appropriate that the acknowledged father of allelopathy, Hans Molisch, should feature in the first article of the inaugural edition of Allelopathy Journal, published in 1994. It is precisely in the allelophatic properties of plants – through their ever-evolving ‘chemical arms race’ - that we have such immense molecular complexity which gives rise to their therapeutical value. Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development Clinical Herbal Studies and Research – A Brief Summary Bai Hua She She Cao, Herba Oldenlandia Oral indigestion of this herb was found to have a dose-dependent effect to enhance macrophage function in vitro and inhibit tumour growth in vivo. Ref.: Wong BY, Lau BH, Jia TY, Wan CP. Oldenlandia diffusa and Scutellaria barbata augment macrophage oxidative burst and inhibit tumour growth. ‘Cancer Biother Radiopharm’ 1996 Feb; 11 (1):5-6 Dong Ling Cao, Herba Rabdosiae Esophageal and cardiac cancer: In one study, 95 patients with middle-tolate stage esophageal and cardiac cancer were treated with preparations of this herb. Preparation I was made by mixing the herb with syrup (1:1 ratio); the treatment protocol was to give 20 to 30 ml three times daily for 2 to 3 months per course of treatment. Preparation II was made by incorporating 4.5 to 5 grams of the herb into pills three times daily for 2 to 3 months per course of treatement. Preparation III was made by mixing 75 to 100 mg of the active ingredient, rubescensine A, in 500 ml of D5W; the treatment protocol was to infuse the preparation IV every other day until a total of 3000 to 3500 mg of compound had been given. The study concluded the rate of effectiveness was 26.7% for Preparation I, 27.6% for Preparation II and 31.8% for Preparation III. Liver cancer: According to one study, 31 patients with primary liver cancer were treated successfully with preparations of this herb. Improved appetite and reduced pain were reported in 80% of the subjects. The survival rate after treatment was 29.6% after 6 months, 12% after one year and 10% after 2 years. Ref.: Xian Dai Zhong Yao Li Xue (Contemporary Pharmacology of Chinese Herbs), 1997; 1468-1469; and He Nan Yi Xue Yuan Xue Bao (Journal fo Henen University of Medicine) 1976; (5):22. Ban Mao, Mylabris Liver cancer: In one study, 300 patients were treated with cantharidin, the active ingredient in Ban Mao, and chemotherapy or radiotherapy. The protocol for cantharidin was 0.25 to 0.5 mg three times daily. The study reported and all-over rate of effectiveness of 65% based on symptomatic evaluation and the tumour size in the liver. In another study, 2 to 6 pills of the herb (each pill contained 0.25 mg of cantharidin, Bai Ji (Rhizoma Bletillae), aluminium hydroxide, and others) were given to 800 patients with liver cancer daily with 45 to 60% showing positive results. There were reductions of symptoms, decrease of tumour size and 12.7% of patients remained alive after one year. Breast tumour: In one clinical study, 42 patients with lobular hypertrophy of the mammary gland were treated with on overall rate of 83% effectiveness using oral, intravenous or intramuscular injection of Tian Men Dong. In another study, 72 patients with mammary cancer showed marked improvement using the same treatment protocols. For oral administration, the protocol was to weigh out 63 grams of the herb, remove the outer layer of the root, add a small quantity of grain-base Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development liquor, steam the preparation for 30 to 60 minutes and ingest the herbs in three equally-divided doses daily. Malignant lymphoma: According to one report, 41 patients were treated with an overall 87.9% effectiveness using an integrated approach of Chinese and Western medicine. Out of 41 patients, 23 received herbal treatment only, and 18 received the combination of herbs and chemotherapy treatments. The herbal treatment consisted of intravenous injection of Tian Men Dong and intramuscular injection of Bai Hua She She Cao (Herba Oldenlandia) twice daily for 3 to 6 months. The injectables contained dosages equivalent in decoction to 10 to 40 grams (with a maximum of 120 grams) for Tian Men Dong and 8 grams for Bai Hua She She Cao. Those who could not tolerate injections were converted to oral ingestion of the herbs three times daily. Recently, these two herbs have been use together in studies to treat fibrocystic breast disorders and cancer of the breast and lymphatic system. Ref.: Yao Xue Tong Bao (Report of Herbology), 1980; 3:2 And: Jiang Su Yi Yao (Jiangsu Journal of Medicine and Herbology); Xing Yi Xue (New Medicine) 1975; 4:193 Dong Chong Xia Cao, Cordyceps Malignant tumours in 30 patients were treated with 1.5 grams of the herb in concoction three times daily over 2 months. The study reported good improvement based on symptomatic assessment in 93% of the patients. Ref.: Shang Hai Zong Yi Yao Za Zhi (Shanghai Journal of Chinese Medicine and Herbology) 1986; 10:25 Da Zao, Fructus Jujubae Several anti-neoplastic components of this herb have demonstrated and inhibitory effect on cancer cells. Continuous administration of mashnic acid, one of the acitive components, for 14 days was more effective than 5-fluorouracil in inhibiting the growth of the cancer cells. Ref.: Ibid. Huang Qi, Radix Astragali In an in vivo study, administration of the herb was associated with the reversal of cyclophosphamide-induced immune suppression. Ref.: Journal of Clinical and Laboratory Immunity, 1988 Mar. 25(3):125 Tian Nan Xing, Rhizoma Arisaematis Cervical cancer: In one study, 105 patients were treated with a 78% rate of effectiveness using both oral and topical applications of the herb. The treatment protocol was to start with an oral decoction starting with 15g/day and gradually increasing to 45g/day. In addition, local topical application was given using various dosage forms such as powder, suppository and injections. Ref.: Zhong Yao Xue (Chinese Herbology) 1998; 1972; 17:8 E Zhu, Rhizoma Curcumae Malignant cancer: Patients with various malignant cancers (stomach, lung, liver, esophageal) were treated with an IV infusion of herbal solution Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development daily. The herbal injection was prepared by mixing 60 to 100 ml of 150% E Zhu solution with 500 ml of D5W. Out of 19 patients in the study, there was complete recovery in 1 case, marked effectiveness in 4 cases, satisfactory improvement in 3 cases and no response in 11 cases. Ref.: Journal of Shandong University School of Medicine; 1980; 1:30 San Leng, Rhizoma Sparganii Hepatic carcinoma: Patients were treated both with herbal injections and powder with a 43.3% rate of effectiveness. Out of 30 patients, marked improvement in 3, good improvement in 10 and no response in 17 was recorded. Malignant tumour: In one study, 31 patients at terminal stage of cancers were treated with a herbal decoction. Marked improvement was seen in 5 patients, good improvement in 19 patients and no response in 7 patients. Ref.: Zhong Liu Yu Fang Yan Jiu (Tumour Prevention Research), 1973, 1:31; Fu Jian Zhong Yi Xue Yuan Xue Bao (Journal of Zhejian University of Medicine) 1983; 3:31 Fu Ling, Poria A herbal preparation was evaluated for its antineoplastic effects in 70 patients in the oncology department of Fouzhou hospital in China. Some patients were given the herbal decoction only, some used it together with chemotherapy, radiotherapy and/or surgery. In all patients, it was observed that the herb strengthens the immune system, improves liver and kidney function, increases appetite and body weight and minimized the adverse side effects associated with chemotherapy. Fen Fang Ji, Radix Stephaniae Tetandrae Lung cancer: Intravenous injections of a herbal preparation (180 to 300 mg of the herb mixed with saline or D5W) were given to 97 patients with promising results. The preparation showed a marked inhibitory effect on DNA and RNA of the cancer cells. Ref.: Journal of Chinese Medicine; 1980; 3:597 Mandarin, Cantonese, Pharmaceutical Name Therapeutic Action/Pharmacological Effects San Leng King Sam Ling, Rhizoma Sparganii Dispels blood stasis and masses, activates chi, carcinoma and malignant tumours, antineoplastic, caution with anticoagulants and anti-platelet drugs* E Zhu Fung Ol Shud, Rhizoma Curcumae Dispels blood stasis and masses, activates chi, antineoplastic, malignant growth, caution with anticoagulants and anti-platelet drugs Shui Zhi Soyi Jah, Hirudo, Leech Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development Dispels blood stasis, anticoagulant, caution: hirudin has same chemical composition as lepirudin (Refludan) * Quan Xie Chuen Kit, Scorpio Eliminates toxins, toxic heat and nodules, opens channels, breaks up tumours, antineoplastic, caution: overdose may occur with intake of 30-60 gr. ** Wu Gong Scolopendra, Centipede Eliminates toxins, toxic heat and nodules, opens channels, breaks up tumours, antineoplastic, caution: Overdose may cause sudden drop in blood pressure & temperature, slow heart beat, dyspnea. At small doses it stimulates cardiac tissue, at large doses it paralyzes cardiac muscle and inhibits respiration. ** Ba Dou Fructus Crotonis, Croton Fruit (Seed) Eliminates cold accumulation, phlegm & water, antineoplastic, best applied topically as past, can be taken internally, but very toxic, max. dosage 0.3 gr., can cause severe toxic shock syndrome! Xuan Shen King Yeun Sum – Radix Scrophulariae Clears heat, toxins nodules & palpable masses, reduces gland swellings, antibiotic Tian Men Dong Radix Asparagi Clears heat, antibiotic, antineoplastic, breast tumours & malignant lymphoma, 20-25 gr. 3 times a day, steamed for 30-60 min. in grain-based liquor, seems to be particularly effective in combination with Bai Hua She She Cao (Herba Oldenlandia) Jiao Gu Lan Rhizoma seu Herba Gynostemmatis Clears heat, toxins, inflammation, antineoplastic, inhibits most types of cancer cell, use for all cancers, may cause drowsiness, sedation and mild stomach discomfort Yan Dan Zi Fructus Bruceae Clears heat & toxins, treats various cancers, use topically to dissolve hardenings, slightly toxic, internally use up to 30 seeds, remove shell, give in pill form due to extreme bitterness, as powder give 2 gr. Ban Mao Mylabris, Blister Beetle Eliminates toxins, dispels nodules & palpable masses, antineoplastic, best applied topically, very toxic, max. dosage 0.03-0.06 gr., very strong stimulation and Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development irritating effect on skin, don’t apply over prolonged periods, caution: do not overdose ** Lu Feng Fang Shan Ci Gu Nidus Vespae (hornet nest) Clears toxins & relieves pain for mastitis and ulcerations with pus, tumour, fibrocystic breast disorder, necrosing nodules, antineoplastic, internally dry-fried & suspended in grain-based liquor, externally as wash or ointment Pseudobulbus Cremastrae Clears heat & toxins, drains abscess & nodules, antineoplastic, tumours and breast carcinoma, slightly toxic, usual dosage 3-6 gr., toxic dose is 15-45 gr. Bai Hua She She Cao Herba Oldenlandia Clears heat, toxins, abscesses, various types of cancer, dose-dependent effect to enhance macrophage function in inhibiting tumour growth, suggested cancer dosage is 60 gr. Zi Cao Gen Bah Gee Jao, Radix Lithospermi Clears heat & toxins, invigorates blood circulation, antineoplastic properties by interfering with DNA sythesis of cancer cells at G2 phase Ban Zhi Lian Boon Chi Lin - Herba Scutellariae Barbatae Clears heat & toxins, tumours and generally antineoplastic properties, enhances macrophage function and inhibits tumour growth, reduces abscesses and swellings * Certain Phytochemicals can cause potentially dangerous herb-drug interactions. Due to their synergistic properties some herbs will either inhibit or greatly increase the bioavailability of an active ingredient in drugs such as in anticoagulants and antiplatelet medicines. Examples for anticoagulants are heparin, warfarin (Coumadin) and enoxaparin (Lovenox); examples for antiplatelet drugs include aspirin, dipyridamole (Persantine) and clopidogrel (Plavix). * * Quan Xie (Chuen Kit, Scorpio), as some other substances, is toxic. In case of overdose, ingest 20 gr. Of Yuan Ming Fen (Natrii Sulfas Praeparata) to eliminate toxins through defecation and prepare detoxing formula with herbs such as Jin Yin Hua (Gam Ngam Fwa, Flos Lornica), Gan Cao (Gum Jao, Rx. Glycyrrhizae), Da Huang (Tai Wong, Rx. et Rhz. Rhei), etc. Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development Table 4: Herbs Commonly Prescribed with Chemotherapy, and Chinese Anticancer Agents Back to the Table of Contents Botanical name and family Traditional indications Huang Qi Radix Astragalus membranaceus (Fisch.) Bge. (Leguminosae) Supplements qi, increases yang, consolidates the surface, increases resistance to disease, controls sweating, delivers fluids, disperses swelling, discharges pus. Increased CD4/CD8 ratio and phagocytic activity in patients with gastric cancer undergoing chemotherapy. Stimulation of lymphocyte IL-2, IL-3, IL-6, TNFa and IFN-g. Diuretic effect and antinephrotoxic effect. Antiinflammatory effect. Hepatoprotective effect. Saponins: astragalosides I-VIII, acetylastragaloside I. Flavones: kaempferol, quercitin, isorhamnetin, rhamnocitin, formononetin, calycosin. Polysaccharides: astragalans I,II,III. Glucans: AG-1 and AG- 2. Bai Zhu Rhizoma Atractylodis macrocephalae Koidz. (Compositae) Replenishes qi and reinforces the spleen. Harmonises the spleen and stomach, relieves fatigue. Induces diuresis and eliminates damp. Arrests excessive perspiration and spontaneous sweating. Increased phagocytosis, lymphocyte transformation, rosette formation, and serum IgG post chemotherapy. Increases body weight and endurance. Potentiates reticuloendothelial system. Diuretic effect, antiulcerative effect, hypoglycaemic effect. Anticoagulant Essential oil:s atractylon. Lactones: atractylenolides II,III. Vitamin A. Sesquiterpene and furfural. Pinyin name Modern research Chemical Constituents Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development effect. Hepatoprotective effect. Lowers blood pressure and dialates blood vessels. Ling Zhi Ganoderma lucidum (Leyss. Ex Fr.) Karst. (Basidiomycetes) Nourishes, tonifies, supplements qi and blood. Removes toxins, astringes essence and disperses accumulation. Relieves fatigue and subdues deficiency insomnia. Antitussive effect. Expectorant effect. Hypotensive effect. Hepatoprotective effect. Antibacterial effect. Sensitises radiation effect. Protects from radiation damage. Immune stimulating effect. Ergosterol, coumarin, mannitol, polysaccharides, organic acids, resins. Dang Shen Radix Codonopsis pilosula (Franch.) Nannf. (Campanulaceae) Tonifies qi, increases body resistance, promotes digestion & absorption of nutrients. Increases secretion of body fluids. Promotes digestion and metabolism. Stimulates the CNS: decreases monoxidase-B (MAO-B) activity in the brain. Hematopeiteic. Hypotensive effect. Significantly decreases erythrocyte electrophoretic time and fibrinogen. Enhances cardiac function and increases tolerance to cold without increasing body weight and it elevates activity of superoxide dismutase (SOD). Increases Phytosterols and triterpenes: spinasterol and Dglucopyranoside, 7-stigmasterol, 5,22stigmasterol, taraxerol, taraxeryl acetate and friedelin. Phenols: syringaldehyde, vanillic acid, syringin, tangshenoside I. Essential oils: methyl palmitate, octadecane, nonadecane, heptadecane, carboxylic acid. Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development phagocytosis. Promotes leukocyte production. Increases haemoglobin levels, antagonises insulin induced hypoglycaemia, but was ineffective with phagocytosis and the transformation of lymphocytes. It also inhibited type II allergic reactions and stimulated the adrenal cortex Inhibits transplanted sarcoma 180 in mice. Fu Ling Sclerotium Poria cocos (Schw.) Wolf (Polyporaceae) Induces diuresis and excretes dampness. Invigorates the spleen function. Tranquillises the mind. Increased monocyte GMCSF production Enhanced recovery of myelosuppression in mice after radiation. Increased spontaneous rosette formation, lymphocyte transformation, and serum IgG. Diuretic effect. Sedative effect. Antitumour promotion effect. Increases cardiac contractility. Polysaccharide: pachyman. Triterpene: pachymic acid, tumulosic acid, eburicoic acid, pinicolic acid. Herba Taraxacum Removes toxic heat. Antimicrobial effect. Immune Taraxasterol, taraxacerin, taraxicin, Antineoplastic agents Pu Gong Ying Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development mongolicum Hand.- Mazz. (Compositae) Removes swelling and nodulation. Relieves dysuria. stimulating effect: choline, inulin and increases pectins. peripheral lymphoblast transformation rate. Choloretic effect and hepatoprotective effect. Jin Yin Hua Flos Lonicera japonica Thunb. (Caprifoliaceae) Removes toxic heat. Dispels wind heat. Antimicrobial effect. Antiinflammatory effect, antilipaemic effect. Decreases pregnancy rate after mating. Antispasmodic effect. Diuretic effect. Shan Ci Gu Bulbus Cremastra variabilis (Blume) Nakai (Orichidaceae) Reduces heat. Removes toxins, disperses accumulation, dissipates swelling. Antineoplastic Tulipine, colchicines. effect. Cardiotonic effect. Antiviral effect. Huang Yao Zi Rhizoma Dioscorea bulbifera L. (Dioscoreaceae) Resolves phlegm. Controls cough. Disperses goitre and controls bleeding. Antibacterial effect. Antifungal effect. Increases uterine contraction. Terpenoids: diosbulbin A,B,C,D. 2,4,6,7tetrahydroxy-9,10dihydrophenanthrene, 2,4,5,6- tetrahydroxyphenanthrene. Tannin. Bai Hua She She Cao Herba Oldenladia diffusa (Willd.) Roxb. (Rubiaceae) Removes toxic damp heat, clears abscesses, infections with fever. Increases phagocytosis, lowers fever, arrests growth of spermatogonia and empties convoluted seminiferoous tubules. Iridoid glycosides: oldenlandosides Aand B. hentriacontane, stigmasterol, ursolic acid, oleanolic acid, b sitosterol, sitosterolDglucoside, p-coumaric acid and flavonoid glycosides. Chlorogenic acid. Inositol and flavonone. Essential oils: 2,6,6trimethyl-2-vinyl-5hydroxytetrahydropyran and linalool. Kirsten Dhar, The Clinic of the College of Chinese Medicine, Research and Development