Preclinical and Clinical Studies of Botanical Agents for Breast Cancer Society for Integrative Oncology Monthly Webinar Series May 16, 2013 Debu Tripathy, M.D. Professor of Medicine University of Southern California Norris Comprehensive Cancer Center Polling Question The following best represents my approach or beliefs about herbal therapy for cancer patients 1. It should generally be discouraged 2. Could have harms, but OK if shared with oncologist 3. Decisions best made by a licensed acupuncturist "Systems" of Medicine in Breast Cancer • Include Chinese, Tibetan, Ayurvedic, Amazonian medicine • Can include of acupuncture, specific diagnosis and herbal formulae, other lifestyle/diet changes • Centuries-long tradition of use and experience • Based on written texts and documentation • Some conformity and reproducibility in practice • Licensed profession in many states (Chinese Medicine, Acupuncture/Herbal Medicine) • Widely used alone or in combination with standard therapy “Many natural substances contain bioactive properties – perhaps due to evolutionary symbiosis” PHA 3-28-.03 Challenges for Botanical Studies • Most utilized forms of herbal therapy are highly individualized and less amenable to controlled clinical trials • Inconsistency of botanical/herbal product • Unknown/unwanted biological effects – Organ toxicity (eg. GI upset, rare renal/hepatic injury) – Drug interactions • Lack of adequate preclinical models (eg. interactions with stromal and immune cells) Drug Herb Interactions Are Common http://www.nlm.nih.gov/medlineplus/druginformation.html Indanavir plasma levels In addition to CYP metabolism enzymes, herbs can affect: 1. Membrane transporters Absorption, excretion, cell efflux 2. Surface and nuclear receptor binding Examples from St. John’s wart (hyperforin) 3. Protein binding / displacement Taken from: Meijerman I et al. Oncologist 2006 and Wang Z et al. Clin Pharmacol Ther 2002 4-Herb Chinese Medicine PHY906: Gastrointestinal Toxicity and Anti-tumor Efficacy Lam W, et al. Sci Transl Med 2010 Clinical Scenarios in Breast Cancer Tested Using Traditional Chinese Medicine • Alleviation of Side Effects of Chemotherapy • Anti-cancer Effects (tumor response) – Single botanical agent – Individualized formulae • Menopausal Symptoms • Breast Cancer Prevention Strategy for Botanical Drug Discovery Herbs or regimens used historically for specific indications Phase I/II In vitro phenotypic safety and studies of herbal efficacy extracts (aqueous) studies Comparative HPLC and other Phase III studies compound isolation strategy Preparation of Aqueous Extracts from Chinese Herbs • Selection of herbs – Source: Shen Nong Herbs, Berkeley,CA. Provided by Isaac Cohen, L.Ac., American Acupuncture Center, Berkeley, CA. – Herbs reputed to have anticancer properties in TCM – Herbs considered to have breast tissue selectivity in TCM • Preparation of herbal extracts – 7.5 g dry, ground herb + 125 ml water – Bring to boil, then simmer 45 min – Decant and centrifuge – Sterile filter – Store at 4oC Inhibitory Activity of Herbal Extracts on Breast Cancer Cells (-) 0-25% (+) 26-50% (++) 51-75% (+++) 76-100% Family Genus species Chinese name SK-BR-3 MCF7 MB-231 BT-474 MCNeuA Anacardiaceae Rhus chinensis wu bei zi --+-+ Araceae Acorus gramineus shi chang pu ---+- Arisaema consanguineum tian nan xing ----- Araliaceae Acanthopanax gracilistylus wu jia pi +- +-++ Gynostematis jiao gu lan ----- Panax ginseng ren shen +- +++- Panax notoginseng san qi ----- Berberidaceae Epimedium grandiflorum yin yang huo +++ + +++ + ++ Boraginaceae Lithospermum erythrorhizon zi cao ---++- Burseraceae Boswellia carterii ru xiang ++ + + + ++ Commiphora myrrha mo yao ++ +++ +++ ++ +++ Campanulaceae Lobelia chinensis ban bian lian ---+- Caprifoliaceae Lonicer japonica jin yin hua ++ - ++ - + Caryophyllaceae Stellaria dichotoma yin chai hu ++-+-- Vaccaria segetalis wang bu liu xing +++ +++ +++ ++ +++ Chloranthaceae Sarcandra glabra jiu jie cha ++ +++ +++ + ++ Compositae Artemisia argyi ai ye ++ +++ +++ ++ +++ Carthamus tinctorius hong hua + - ++ - ++ Rhaponticum uniflorum lou lu +++++- ++ Taraxacum mongolici pu gong ying ++ ++ +++ + ++ Corydiidae Eupolyphaga sinensis (zool.) zhe chong --+-- Cucurbitaceae Luffa cylindrica si gua lou +- - +- Trichosanthes kirilowii (root) tian hua fen ----+ Trichosanthes kirilowii (fruit) gua lou ++ ++ ++ ++ ++ Campbell et al Anticancer Res 2002 Inhibitory Activity of Herbal Extracts on Breast Cancer Cells (-) 0-25% (+) 26-50% (++) 51-75% (+++) 76-100% Family Genus species Chinese name SK-BR-3 MCF7 MB-231 BT-474 MCNeuA Dioscoreaceae Dioscorea bulbifera huang yao zi ++++++ Gramineae Imperata cylindrico mao gen ---+- Semen coicis yi-yi-ren ----- Labiatae Prunella vulgaris xia ku cao +++++ Salvia chinensis shi jian chuan ++ +++ +++ - ++ Scutellaria barbata ban zhi lian ++ + ++ + +++ Laminariaceae Laminaria japonica kun bu --+-- Lardizabalaceae Caulis mutong mu tong ++ + + - + Leguminosae Astragalus membrabaceus huang qi ----- Gleditsia sinensis zao jiao ci +++ +++ +++ +++ ++ Glycyrrhiza uralensis gan cao ++ - - + - Phaseolus calcaratus chi xiao dou ----- Psoralea corylifolia bu gu zhi ----- Pueraria lobata ge gen ----- Saphora japonica huai hua --+-+ Sophora flavescens ku shen ++- - - Sophora tonkineenis shan dou gen ----- Lilaceae Asparagus cochinchinensis tian men dong ----- Anemarrhena asphodeloides zhi mu +++ +++ +++ ++ +++ Fritillaria cirrhosa chuan bei mu ----- Fritillaria thunbergii zhe bei mu ----- Loranthaceae Viscum coloratum sang ji sheng +++ + ++ - +++ Malvaceae Hibiscus mutabilis fu rong ye ---+- Menispermaceae Stephania tetrandra han fang ji ----- Campbell et al Anticancer Res 2002 Inhibitory Activity of Herbal Extracts on Breast Cancer Cells (-) 0-25% (+) 26-50% (++) 51-75% (+++) 76-100% Family Genus species Chinese name SK-BR-3 MCF7 MB-231 BT-474 MCNeuA Oleaceae Ligustrum lucidum nu zhen zi +++ +++ +++ + +++ Orchidaceae Cremastra variabilis shan ci gu ----- Ostreidae Ostrea gigas (zool.) mu li ----- Palmae Daemonorops draco xue jie -+--- Polygonaceae Rheum palmatum da huang ++ +++ ++ - +++ Ranunculaceae Radix clematidis wei ling xian +- - +- Semiaquilegia adoxoides tian kui zi ----- Ranunculus ternatus mao zhua cao ---++ Rosaceae Agrimonia pilosa xian he cao ++++-+ Duchesnea indica she-mei +++ +++ +++ + +++ Eriobotrya japonica pi pa ye +++ - ++ ++ +++ Rubiaceae Heydyotis diffusa bai hua she she cao -++-+ Rubia cordifolia qian cao gen +++ +++ +++ ++ +++ Uncaria rhychophylla gou teng ++ ++ +++ - +++ Salicaceae Salix babylonica liu zhi +++- - Sargassum Sargassum pallidum hai zao ----+ Solanaceae Solanum lyratum shu yang quan --+-+ Solanum melongena qie gen --+-- Solanum nigrum long kui ---+- Sparganiaceae Sparganium stoloniferum san leng --+-- Tremellaceae Tremella fuciformis bai mu er ----- Zingiberaceae Curcuma longa yu jin ----- Curcuma zedoaria e zhu ----- Campbell et al Anticancer Res 2002 HPLC Chromatogram Showing Flavones: Active Compounds from Scutellaria Barbata Isoscutellarein Luteolin Scutellarein Apigenin BZL101 Batch to Batch Consistency Necessary for FDA Regulatory Requirements Drug Substance Lot A-Red and Drug Substance Lot B-Blue TIC of - Q1: from Sample 60 (60-4) of JT_07Oct10_MF_BZL_powder_fi ngerprint_10mg-d1.wiff (Turbo Spray)Max. 2.5e9 cps. 0.17 2.4e9 0.97 2.2e9 2.0e9 27. 81 28. 90 1.8e9 27.30 1.6e9 1.4e9 7. 97 26.21 25.46 1.2e9 24.60 6.76 1.0e9 8.0e8 3.10 3.77 6.0e8 4.0e8 12. 18 2.0e8 0.0 246810121416182022242628 Ti me, min BZL101: Active Compounds Compound Structures Bionovo Name MW formula CAS# Structure Number BNAC5501 Acteoside 624 C H O 61276-17-3 OH 29 36 15 HO O HO O O HO O OH OH HO O O OH OH OH HO OH BNAC5502 Scutellarin 462 C21H18O12 27740-01-8 HO O O O OH O HO OH O BNAC5503 Scutellarein 286 C H O 529-53-3 HO O 15 10 6 OH HO OH O BNER1119 Apigenin 270 C15H10O5 520-36-5 5,7,4’- HO O OH trihydroxyflavone OH O OH BNER1114 Luteolin 286 C15H10O6 491-70-3 HO O 5,7,3’,4’- OH tetrahydroxyflavone OH O Compound Structures, Cont. BNAC5504 Isoscutellarein 286 C15H10O6 5,7,8,4’- tetrahydroxyflavone HO O BNAC5505 Carthamidin 288 C15H12O6 479-54-9 OH – R and S enantiomers HO OH O OH BNAC5506 Isocarthamidin 288 C H O 2569-76-8 HO O 15 12 6 OH – R and S enantiomers OH O BNAC5507 Isoscutellarein-7-O- 462 C21H18O12 glucuronide BNAC5508 Carthamidin-7-O- 464 C21H20O12 glucuronide – R and S diasteriomers BNAC5509 Isocarthamidin-7-O- 464 C21H20O12 glucuronide – R and S diasteriomers BNAC5510 Apigenin-5-O- 432 C21H20O10 glucopyranoside BNAC5511 Apigenin-7-O- 446 C21H18O11 glucuronide BZL-101- Induction of ROS UT UT BZL BZL 100 101 102 103 104 100 101 102 103 104 FL1-H FL1-H MDA MB 231 MCF10A BZL-101 induces significant increase in the levels of ROS in breast cancer cells but not in non-transformed cells Does BZL-101-induced ROS cause DNA damage? BZL101: Selectivity for Cancer Cells • BZL101 inhibits cancer cell lines derived from various tissues • BZL101 does not inhibit normal breast epithelium (HuMEC) • BZL101 does not inhibit normal fibroblasts (IMR90) lung pancreas prostate breast A549 LLC Panc-1 Panc02 PC-3 LNCaP MCF-7 MCNeuA HuMEC + ++ + ++ + + ++ ++ - IC50 1424 492 1054 594 1035 1516 818 619 DNA fragmentation induced by BZL101 40 - < 50% inhibition 35 30 + 51-75% inhibition 25 UT 20 1d ++ >75% inhibition 15 10 2d * IC50 values (ng/ml, concentration at 5 which 50% growth inhibition is seen) 0 % cells with fragmented DNA fragmented with % cells MDA SKBr BT474 IMR90 MB468 Fong S et al. Cancer Biol Ther 2008 6oi r (moa i (1en a 29p l 5%dia 2 %)enr f1 )gt o srti BZL101 Phase 1A - Response Results rT teha r ael