1. Predicting Receptor targets of Traditional Chinese Medicines and Small Molecule TCM Database J. Jia and Y.Z. Chen Department of Pharmacy National University of Singapore Tel: 65-6616-6877; Email: phacyz@nus.edu.sg ; Web: http://bidd.nus.edu.sg phacyz@nus.edu.sghttp://bidd.nus.edu.sgphacyz@nus.edu.sghttp://bidd.nus.edu.sgContent TCM mechanism, targets, roles of bioinformatics and modelingTCM mechanism, targets, roles of bioinformatics and modeling TCM and target databasesTCM and target databases Target prediction by INVDOCKTarget prediction by INVDOCK Bioinformatics study of mechanism of interior warming herbsBioinformatics study of mechanism of interior warming herbs

2. Therapeutics from Traditional Medicines  Potentially novel therapeutic approaches  Existing markets in certain countries and potential market in others  Need for standardization, validation, & further development  Markers for standardization  Molecular mechanism  Safety and efficacy tests  New recipes, combination drug mimics Pharmacology & Therapeutics 2000, 86:191-198

3. Why receptor targets? Therapeutics from Traditional Medicines  Multiple herb therapies  Collective synergistic actions, maintenance and balance.  Mutual accentuation mutual enhancement mutual counteraction mutual suppression mutual antagonism mutual incompatibility  Multiple targets:  therapeutic effects  symptom treatment  toxicity modulation  Immune regulation  drug delivery  energy metabolism  harmonization Pharmacology & Therapeutics 2000, 86:191-198

4. Computer Analysis of Molecular Mechanism of Traditional Medicine Y.Z. Chen and D.G. Zhi Proteins 2001;43: 217 Traditional Medicine Database  Herbal Ingredient & Content Database  Therapeutic Target Database   Drug Adverse Reaction Target Database   Drug Absorption Distribution Metabolism Excretion Database   Computer Match-Making Software  Herbal Ingredient Therapeutic Target Toxicity Target ADME protein  Collective therapeutic and maintenance effects  Toxicity / side effects and modulation  Drug delivery and clearance Matched Targets Mutual Enhancement ? Mutual Counteraction ? Maintenance or Balance ? Delivery or Clearance ?

5. Medicinal Herb Databases at BIDD Comparison with existing TCM databases: Formula: TCM-ID: 1000 TCHFL: 270 Herb: TCM-ID: 1200 TCSHL: 520 TCMD: 1500 Compound: TCM-ID: 9000 CNPD: 3000 TCMD: 6800 TCM Formula Herb Compound Protein Function Structure TCMD TCHF Library CNPD TCSH Library TCM-ID: Traditional Chinese Medicine - Information Database   Only database providing integrated and comprehensive info about: TCM formula, constituent herbs, herbal ingredients, effect on proteins Molecular structure Function at the formula, herb and compound levels

6. INVDOCK Strategy for Match Making Science 1992;257: 1078 Proteins 1999; 36:1

7. INVDOCK Test on INVDOCK Test on Targets of Chinese Medicinal Herbal Ingredients (Am. J. Chin. Med. 2002, 30, 139) Chinese Natural Product Number of Identified Putative and Known Therapeutic Targets Number Confirmed or Implicated Therapeutic Targets by experiment Number of Identified Putative and Known Toxicity/Side effect Targets Number Confirmed or Implicated Toxicity/Side Effect Targets by experiment Acronycine314- Allicin5211 Baicalin1446- Catechin17125- Camptothecine9632 Dicoumarin7131 Emodin6351 Genistin227121

8. INVDOCK Test on TCM Target Prediction INVDOCK Test on TCM Target Prediction Therapeutic targets of Camptothecine (Am. J. Chin. Med. 2002, 30, 139) PDBProteinExperimental FindingTarget Status Theraputic EffectRef 1adsAldose Reductase Diabetes treatment 2gssGlutathione S- Transferase p1-1 Increases intracellular glutathione ImplicatedEnhance radical scavenging activities that may useful in cancer treatment Matsumoto 7iceDNA Polymerase Beta Anti-cancer 1a25Protein Kinase CInhibitorConfirmedInduction of apoptosis in tumor. Martelli Nieve-Neira 1cdkCAMP-Dependent Protein Kinase Anti-cancer 3bctBeta-CateninInduces apoptosis in leukemic cells. 1dviCalpainInhibition of calpain activities.Anti-cancerEymin 1yfoReceptor Protein Tyrosine Phosphatase Causes elevation of PTPase in the cytosol and the nucleus which play a critical role in the induction of the differentiation of IW32 erythroleukemia cells. Anti-cancerWang M 1a35Topoisomerase IInhibitorWang MC

9. INVDOCK Test on TCM Target Prediction INVDOCK Test on TCM Target Prediction Toxicity and side effect targets of Camptothecine (Am. J. Chin. Med. 2002, 30, 139) PDBProteinExperimental Finding Target Status Toxicity/Side Effect Ref. 2cljAcetylcholinesteraseReversible inhibition ConfirmedCauses Cholinergic ToxicityDodds 1aqbRetinol-Binding Protein Interfering with retinol transport that may cause complication in cancer and cardiovascular disease 5rlaArginaseIncrease blood urea level Implicate d Hyperammonemia that may cause nerve toxicity Yu ZJ

10. Chinese Medicinal Herbs All Other Herbs All ingredients of the other herbs Interior Warming Herbs All ingredients of interior warming herbs Exclusive ingredients COMPARE Bioinformatics Study of Mechanism of TCM Herbs Interior Warming Herbs as an Example Pharmacological effects possibly linked to interior warming Other Pharmacological and regulatory effects Interior warming effects primarily come from these ingredients

11. Pharmacological Effects Possibly Linked to Interior Warming

12. Vasodilation and its Relation to Interior Warming Common mode of action (except Hua Jiao) –Inhibit Ca 2+ influx Other Modes of actions –Increase NO –Increase cGMP –Activate TRPV1 –Alpha-adrenoceptor blockade Why vasodiation produces warming effect or sensation? –Blushing A result of vasodilation Warm feeling –Cold extremeties due to loss of beta-adrenoceptor mediated vasodilation –Increased lumen size of blood vessels More blood brought to vasodilated area  Warm sensation 1 Rang HP, Dale MM, Ritter JM, P.K. M. Pharmacology. 5th ed. Edinburgh ; New York: Churchill Livingstone, 2003

13. Pharmacological Effects Possibly Linked to Interior Warming

14. TRPV1 Agonism and its Relation to Interior Warming Transient receptor potential ion channel of vanilloid type 1 –Highly expressed in a subset of primary sensory neurons of the trigeminal, vagal and dorsal root ganglia –Non-neuronal cells 1 1 Geppetti P, Materazzi S, Nicoletti P. Eur J Pharmacol 2006;533(1-3):207-14. Activation leads to many effects –Burning sensation –Vasodilation via release of Calcitonin gene related peptide (CGRP) –Inhibit gastric leisions –Thermogenesis Piperine, 6-gingerol, 10-gingerol, 6-shogaol Associated with vasoconstriction

15. TRPV1 Agonism and its Relation to Interior Warming Dual actions leading to the same warming effect or sensation? 1.TRPV1 causes both vasodilation and vasoconstriction under different conditions 1 –Directly induce vascular smooth muscle contraction, linked to thermogenesis that is abolished by vasodilators –But release neuropeptides e.g. CGRP that causes vasodilation 1 Eldershaw T, Colquhoun E, Bennett K et al. Life Sci 1994;55(5):389-97. VasodilationThermogenesis Bi BaBisabolenePiperine (Abolished by Bisabolene?) Gan Jiang 10-gingerol (Net-effect?) Effect Herb 6-shogaol6-gingerol 6-shogaol6-gingerol 2.TRPV1 agonists reported to result in hypothermia due to response to thermogenesis and activation of warm sensors 1, –False sense of warmth –Not truly ‘therapeutic’

16. Acknowledgement Current Group Members: Computer-Aided Drug Design: CY Ung, XH Ma, XH Liu, Pankaj Kumar, F Zhu, X Liu, J Jia Protein Function, Interaction, Network: HL Zhang, CY Ung, XH Ma, F Zhu, WK Teo, Z Shi Databases and Servers: J Jia Medicinal Herb: CY Ung, Pankaj Kumar, Cao Jinyi(undergraduate students) Microarray and biomarkers: J Jia, ZQ Tang Former Members: PhD: ZW Cao (Prof SCBIT, Tongji U), ZL Ji (Assoc Prof Xiamen U), X Chen (Assoc Prof Zhejiang U), CW Yap (Assist Prof NUS), LY Han (Postdoc NIH), CJ Zheng (Postdoc NIH), HH Lin (Postdoc Harvard ), J Cui (Postdoc U Georgia), H Li (Postdoc Einstein College Med) Research Fellow/Assistant: ZR Li (Assoc Prof SiChuan U), Y Xue (Prof SiChuan U), W Liu (Assoc Prof DUT), D Mi (Assoc Prof DUT), CZ Cai (Prof ChongQing U), DG Zhi (Postdoc, Berkeley), MSc: Y.J. Guo (Postdoc NIH), L.Z. Sun (RA, U Tenn.), J. F. Wang (MSU), L.X. Yao (Columbia), S Ong (Washington U), H Zhou (local company), B Xie (local company) BSc: W.K. Yeo (IMCB, Novartis)