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DCC_Cambridge_120109final by xiaoyounan

VIEWS: 4 PAGES: 17

									Profiling of Matrix Metalloprotease
   Inhibitors without Enzymes




               13th January 2009
        DDC Network Meeting in Cambridge

                 Vera Janowski
             PhD advisor: Kay Severin      Marie Curie Research Training
                                                  Network - DCC
                 From Bonn via Münster
                     to Lausanne                                  Marie Curie Research Training
                                                                         Network - DCC

Bonn 1982-2002




                                                                         Thesis at the UCSB
                                                                                (2007)

                                                 Lausanne 2007-
Münster 2003-2006




                                     2002-2003                                 Erasmus in
                                     IU Bremen                                  Zaragoza
Diploma in Chemistry 2007                                                        (2005)
                                                                                          2
Diploma in Business Chemistry 2008
Laboratory of Supra-
molecular Chemistry                Marie Curie Research Training
                                          Network - DCC




        •   Superior research facilities and equipment
            (NMR, UV-vis micro plate reader, fluorescence
            plate reader...)
        •   Weekly group meetings and excellent
            mentoring
        •   Many interesting conferences and seminars
                                                           3
             Training and Teaching                           Marie Curie Research Training
                                                                    Network - DCC

•   Excellent PhD and Master courses, e.g.:
    Current Topics in Mass Spectrometry,
    Inorganic Chemistry
    (Fundamentals and Properties),
    Biological Chemistry
    (Introduction to the molecular and chemical basis of disease and therapy)
•   Inorganic Chemistry Summer School in Villars-sur-Ollon:
    The Challenge of Future Energy Sources
•   Swiss Chemical Society Snow Symposium in Fiesch
•   World Knowledge Dialogue in Crans-Montana:
     - Collective network knowledge & human individual intelligence
     - Collaborative Behavior, Altruism & Conflict
•   Chirality Conference in Geneva, including a course on NMR Spectroscopy
    for Determining Molecular Stereochemistry and Enantiomeric Purity
•   Interesting Training courses within the Marie Curie Network

•   Supervision of a Master project student
•   Teaching of 1st year Biology and 3rd year Chemistry students                     4
                     Network Meetings                           Marie Curie Research Training
                                                                       Network - DCC




    presentation of results




•     regular meetings enable to understand the projects of the other PhDs in the
      network well  fruitful discussions, new ideas
•     different backgrounds offer a new perspective on problems
•     presentation of results are a good preparation of conferences and together
      with the progress log offer a way to reflect one`s work                    5
                                  Matrix
                             Metalloproteases                                 Marie Curie Research Training

                                 (MMPs)                                              Network - DCC




                                                                • 25 different isoforms
                                                                • Zinc-dependent endopeptidases
                                                                • Zinc bound by three histidines
                                                                • Associated with tissue repair,
                                                                  arthritis, metastasis
                                                                • Only one Inhibitor clinically
                                                                  approved




MMP 3
Gilian Murphy, Vera Knäuper, Susan Atkinson, George
Butler, William English, Mike Hutton, Jan Stracke, Ian Clark,
Arthritis Res, 2002, 4, S39-S49.                                                                      6
                       MMP Inhibitor
                        Synthesis                                                      Marie Curie Research Training
                                                                                              Network - DCC

                                                                                                  NR'R'
                                                                                                  '


     HO                                                                        HO
                        O


               +                +      HNR'R''          (Betti/Mannich reaction)
                   H        H
      O    N                                                                       O          N
           H                                                                                  H
                                R', R'' = H, Me, benzyl...




• Easy to synthesize and good to recrystallize
• But some not soluble in H2O at pH 7.4 (DMSO content needs to be
  as low as possible for the enzyme assays)
• Synthesis of structurally more diverse inhibitors failed (primary
  amines + halogen atom substituted ones)
• Slow oxidation forms very colored products (only small changes in
  the NMR spectrum after 1 day)  should be able to be
  circumvented by using methyl substituted dihydroxypyridine
                                                                                                               7
                                    MMP Inhibitors                                                                                     Marie Curie Research Training
                                                                                                                                              Network - DCC
                                O                                      H
                                                                       N
                                         N
                     OH   HO
                                                                                        OH                                                        N

                                                 OH                                                                      O

                                                                                                                                                            OH
        N            OH                                                                                   N
                                                                           N            OH
                                             N   OH
                          O                                                                                                              N
                                                                                                                   OH
                                                      N                                                                                               N     OH
                                    OH
            N                                                                  N                                                             OH

N                                                                                                 N                OH

                           N        OH                        HO               OH
                           H                                                                                                       N         OH
            OH
                                                                                                                        OH

                                O

    N       OH        N                                                        N
                                    O                                                                 N                 OH              N


                           OH
                                                                                    N                                                        OH

                                                                   N                                          N

                 N         OH
                                         N                                                                                         N         OH
                                                                                             HO               OH
                                N
                                                                                                                             NH2
                                                          N


                                    OH
                                                                                                                                   OH
                                                                               OH

                                                                                                                                                                 8
                           N        OH
                                                                                                                    N              OH
                                                                   N           OH
                  Concept            Marie Curie Research Training
                                            Network - DCC




              +                     Correlation
                                     “factor”
“Biological         “Chemical
 system”             system”

                                  Prediction of IC50
                                 values for different
                   Correlation
              +                        Matrix-
                    “factor”
                                  metalloproteases
“Chemical
 system”                              (MMPs)


                                                             9
           Indicator Displacement
                Assays (IDA)                                       Marie Curie Research Training
                                                                          Network - DCC




                                                                        Indicator
                               Analyte
              Indicator                               Analyte



             Receptor                                Receptor

                          Review: E. V. Anslyn et al., Acc. Chem. Res., 2001, 34, 963.


• Non-covalent binding
• Free and bound indicator differ in optical properties
   absorption (colorimetric sensor) or emission (fluorescent sensor)




                                                                                         10
                 Multicomponent
             Indicator Displacement                     Marie Curie Research Training

                 Assays (MIDA)                                 Network - DCC




                                            Indicator
                                           Indicator        Indicator
                                                           Indicator


                         Indicator
         Receptor                          Receptor        Receptor



                    +                       Indicator      Indicator
                                                           Indicator
                                           Indicator
                         Indicator


         Receptor                           Receptor       Receptor




• Multiple indicators/receptors with different absorption/emission
   Dynamic combinatorial library
   big dynamic range


                                                                              11
      Multivariate Analysis                Marie Curie Research Training
                                                  Network - DCC




                                       Analyte 1
               Linear
Analyte 1
            Discriminant
              Analysis
                                                Analyte 3
Analyte 2                  Analyte 2




Analyte 3




                                                                 12
        IC50 Determination       Marie Curie Research Training
                                        Network - DCC



N



         OH




    N    OH




                    25



                    20



                    15



                    10



                     5



                     0
                         1   2   3        4        5

                                                       13
Marie Curie Research Training
       Network - DCC
Marie Curie Research Training
       Network - DCC
             Further Work              Marie Curie Research Training
                                              Network - DCC


• Complete the synthesis of new inhibitors using a
  methyl substituted dihydroxypyridine as starting
  material for the Mannich reaction
• Determine IC50 values of the new inhibitors
• Optimization of the chemical system (MIDAs)
• Do the computational analyses




                                                             16
       Acknowledgements   Marie Curie Research Training
                                 Network - DCC




Prof. Kay Severin

Severin Group

Marie Curie Network


                                                17

								
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