1、 Wei DUANSchool of MedicineDeakin UniversityMelbourne, AustraliaMolecular Medicine for Heart FailureIsozyme-specific Modulation of PKC Heart Failure in AustraliaThe prevalence of heart failure in Australia is 6.3% .The heart failure prevalence is 10% in people aged over 65 years (this population wil

2、l double over the next 50 years in Australia). Stages of heart failure from Goldman: Cecil Medicine, 23rd ed Effusion from congestive heart failure from Mason: Murray & Nadels Textbook of Respiratory Medicine, 4th ed2Molecular Cell Biology underlying heart failure:PKC Activation and Heart FailureCir

3、culation Research. 2007;101:195 3J. Clin. Invest., 115:527,2005Activation of PKC and Heart FailurePKC is activated by Gq/G11 -PLC -IP3-SR-released Ca+ and DAGSarcoplasmic Reticulum)4wo5Nature Medicine, 10:239,2004Protein Phosphatase-1Phospholamban P ca+ uptake, increased Ca+ release & contractile fo

4、rce)Sarcoplasmic reticular calcium uptake proteinPKCa Ca+ cyclingReduced1-contraction strength2ralaxation3Cardiac reservePKCa is activated by Ca+, stress agonists, pressure load, myocardial infarction, angiotensin, Transition in failing hearts fromadequate compensation To advanced failureProtein Pho

5、sphatase Inhibitor-1Calcium handling proteins:PLB, SERCA-2a6Domain Structure of PKC7Study of the function of C-terminus of PKC-alpha10The last 10 amino acid residues are essential for the catalytic competence of PKC-alphaIntrinsic activity, protamine sulfate as substrate.DAG- activation Histone H1 a

6、s substrateS.S. Yeong, et al., J. Biol. Chem. 281:30768, 200611The critical role of the very C-terminus of PKC-a in the activation of MAPK S.S. Yeong, et al., J. Biol. Chem. 281:30768, 200612Aptamers (from Latin aptus, means “fitting”)Synthetic RNA, DNA or peptide that binds to protein targets with

7、high specificity and affinity-Fold into well-defined 3-D structure-Bind by complementary shape interactions-Not toxic or immunogenic-Effectively inhibit the function of target protein-No prior knowledge of the 3-D structure of the target is requiredthrombinCurrent Opinion in Chemical Biology, 9:336-

8、342, 200516Nucleic Acids Base PairingtRNA17SELEX: Systemic Evolution of Ligands by Exponential enrichment.18Develop Aptamers Targeting the very C-terminus of PKC C-terminus of PKC is exposed (accessible) from IP experiments19Source of Chemical LibrariesCommercial sources (free of royalty)-NCI (/docs

9、/misc/available_samples/dtp_indsamples.html)Structural Diversity Set, version 1 (1,991 compounds) Structural Diversity Set, version 2 (1,986 compounds) Mechanistic Diversity Set (879 compounds) Open Collection 1 (90,000 compounds) Open Collection 2 (10,000 compounds)-ChemDiv ()-ChemBridge Corporatio

10、n (/chembridge/)Partnership with pharmasPartnership with academia MOLECULAR LIBRARIES SCREENING CENTERS NETWORK (Established in 2004) 9 Centers Sources of Current or Off-patient medications-Prestwick Chemical Co. (1120 off-patient drugs)-Microsources: The Spectrum Collection (2000)-Sequoia (a large

11、collection)-NIH Brain Bioactive Compound collection21Aptamer-guided Heart-Specific Delivery22Molecular Medicine for Heart DiseasesEngineering nanoparticlesDrug loadingTargetingControlled Release (heat, magnetic field, pH), engineering the nanoparticle system such that the intracellular drug-release kinetics is greater than the drug-efflux kinet