Proposal for research at the Sunderland Pharmacy School by nyut545e2


									Anti-Cancer drugs -
     Telomeres and Cell Senescence
 •   Chromosomes within animal and                Cartoon representation of
     human cells have a group of guanine          Guanine rich
     groups (telomeres) at their 3’termini.       Hoogsteen base paired

 •   These form a 4-membered structure
     or G-tetrads. Upon cell division a
     number of these guanine groups are
     lost and typically not replaced.         Unzipping of chromosonal DNA
     Eventually the number of Guanine
     groups is reduced to the point that
     other cell-defence mechanisms
     destroy the cell (apoptosis) to reduce
     the risk of mutation of the gene
     sequence which may lead to disease,        Imperfect DNA replication
     including certain cancers.                 results in loss of telomere
                                                G-tetrads shortening telomeres
                                                in daughter DNA strand
                   G-Quadruplex Ligands
•   The natural shortening of telomeres in eukaryotic cells has been implicated in
    senescence and subsequent apoptosis.
•   The competing action of telomerase is restricted to certain tumor and early
    stem cells.
•   The genetic coding for the telomerase enzyme and the potential inhibition of
    telomerase expression are considered important areas of research, not merely
    from the perspective of understanding the genetic dependency but, via a new
    fundamental maxim, will provide insight into prospective treatments.

       N    N      H   N       N        N

       N               N
                   O H                  N
           N                            H
               H                O
     HN                                  NH
               O                    N
       N               O                N
                   N H

       N       N       N   H    N       N
     Telomerase and telomere extension
•   Tumor cells are the exception to this rule,
    in that an enzyme called Telomerase is
    produced and goes about replacing the G-
    quadruplex structure on the ends of the         Imperfect DNA replication mechanisms
                                                    results in loss of telomere G-tetrads shortening
    chromosomes making them theoretically           telomeres in daughter DNA strand

•   A way of stopping the replenishment of the
                                                  Telomerase protein comprising RNA and primer
    guanine groups would be to formulate a
    chemical that would adhere to the G-
    quadruplex in such a way as to prevent this
    telomeric elongation.

•   This would cause cancer cells to lose their
    telomeres as a normal healthy cell does.
    The fact that cancer cells replicate faster
    than normal healthy cells (whose
    telomeres are longer anyway) would serve
    to selectively diminish the length of
    telomeres on chromosones in cancerous
    cells and result in them dying (via
    apoptosis) faster than healthy cells.
                     Current thinking
•   Antisense oligonucleotides
•   Telomerase antagonists
•   Oligopeptide vaccines
•   G-Quadruplex Ligands
     – Braco19 (Antisoma)
     – Xenograft tumour models clinical trials
                                                                                                •   One of the strategies would be to design
                                                                                                    a molecule capable of mimicking the G-
                                                                                                    quadruplex structure.

                                          N            N
                                                                   H                            •   High throughput screening using for
                                                                   N        N
                                                                                            N       example DOSY NMR would allow the
                                                                                        N           formation of both static and dynamic
G-quadruplex model
                                      N       O                        O    N
                                                                                                    combinatorial libraries of small organic
                                              O            N                    N                   species ultimately capable of stabilising
                                  N                            N            N
                                                                                                    the biological equivalent.
                                                                                                •   Significant research could be carried out
                                                                                                    in the mode of drug action and the
 Linker Group
                                                                                                    actions of the potential inhibitor within
                                                           O                    N
                                                                                                    living systems.
                             N                                         HN           O
                                                                                                •   The investigation of potential G-
                                                                                                    quadruplex liganding agents would also
                                                                                                    involve consideration of physicochemical
  Calix foundation                                                                                  aspects of both the ligand library and
                                                                                                    the biological systems affected, inculding
                                                                                                    thermodynamics and chemical kinetics.
                   Research Calix[4]arenes in
                     host-guest chemistry
                                                          O                           O
                                    O                                   O         O
                                                              NH                 HN
                                        NH                    HN                      NH

                                                                    O            HN    O
           I              I             HN   O            O        NH                 HN   O
            I             I

                              Et3N, 15% Pd(OAc)2, DPPP,
                OBu BuO
                              DMF, 100oC, 275hrs
                                                                            OR   RO
                OBu BuO
                                                                        OR       RO

On the scope and limitations of the Heck reaction of upper rim tetraiodocalix[4]arenes
J. Chem. Soc. Perkin Trans. 1, 2001, 24, 3393-3398, Kuhnert N and Le Gresley A
Capusle formation and binding to Pesticide
       8.1 Å

                                                                S       S
                                                                            S       S

                                            13.1 Å            fungicide tetramethyl-

 Further corroborated by the ESI-MS showing a signal at m/z 2898

 Synthesis and capsule formation of upper rim substituted tetra acrylamido
 calix[4]arenes Organic and Biomolecular Chemistry, 2005, 11, 2175-
 2182 N. Kuhnert and A. Le Gresley
    Dynamic combinatorial libraries using
                                                                                                OMe                                           OMe

        O            O
                          O                                                   N            N                                N            N
            O                                                                                      N                                             N
                                                                                  N                                             N

            O                 O
                                           selection of amines
    O                    O                                                        O                            OR               O
                                                                          O                    O       O                O                    O       O
O       O                     O   O
                                                                      O       O                        O   O        O       O                        O   O

                                      HN       NH

                OR                    H         H
                     RO                                                                                                             OR
                                                                 OH                   OR   RO                                            RO
                OR   RO
                                           S                                          OR   RO                                       OR   RO
                                                    Biotin   O

                 The synthesis of static and dynamic combinatorial libraries using deep
                 cavity tetra-formyl calix[4]arenes N. Kuhnert and A. Le-Gresley,
                 Tetrahedron Lett. 2005,46, 2059-2062.
         Heck Methodology
                                                R'O O            R'O     O
                                               O OR'                   R'O   O
I                I
I                I
                         Et3N/ 10%
                         Pd(OAc)2/ DPPP

    OR   RO                                             OR       RO
                         DMF 100oC 24-150hrs
    OR   RO                 O                           OR       RO


          R = n-Bu

          R' =
                     2                    3                  4

                            O                                     O

                     5                    6                  7
                       Reversible Reactions
               O                        O                                         R' N
                                                                                   R' N                                   N R'
                   O                        O                                                                               N R'

                    O O             O O                                                     O O                   O O
                                                            MS 4A                          O O                      O O
                   O O                O O

                         OR RO                                                                         OR RO
                         OR RO                                                                         OR RO

                           5                                                                               8-22

               R' =                                              OH


                       a            b       c           d                     e                    f                  g
                                                                                  O            HN                 N
                       H3C (CH2)7                                                 O                                        N
                                                                                               O       N              N
                       h                        i                             j                k                      l
                                                                C O

                                                    O                 O
                                                            m             n                o

Kuhnert, N and Le-Gresley, A. Synthesis of upper rim calix[4]arene carcerands.
Tetrahedron Letters, 98, 1274-1276. 2008
                       Combinatorial Library

                                     O   23
                                HN       NH
                            O                 O                                                                                   N
                                                                                     CHO                                    CHO
                       Barbituric Acid                        OHC                                OHC
                                                                        O O        O O                        O O         O O
                                                                                           and            O    O            O O
                                                                    O                O O

                5cgg        O
                                         24                               OR RO                                 OR RO
                       HN           NH
                                                                          OR RO                                 OR RO
                       H             H
                                                  OH                          5c                                    5ae

                     Molecular                                                        Intensity
        Imine                                          Mass Peak (m/z)                                               Guest
                     Formula                                                          (relative %)

        5c           C91H87O15N                        1434                           35                             None
        5ae          C101H96O15N3                      1590                           15                             -
        5a           C91H87O16N                        1450                           10                             -
        5e           C94H90O15N2                       1487                           5                              -
        5e           C84H90O15N2                       1487                           39                             23
        5cgg         C99H105O13N3                      1540                           25                             24

Le Gresley, A The design and synthesis of deep cavity calix[4]arenes in the development
of static and dynamic macrocyclic libraries. European Journal of Organic Chemistry, in
press. 2009
   Anthracene Diacrylamides

                                               NR2        O       NR2    O

                      O         Ligand
           +               DMF Pd(OAc)2 Et3N

  X                                               X

X=Cl            a. R= Me
X=Br            b. R= H                                           NR2    O
X=CF3SO3        c. (CH2CH2)2O
                                               1R=H                2R=H
                                               3 R = Me            4 R = Me

                                               5R=                 6R=

                                                          O                   O
                                HETCOR & Activity
                      C2         C1        C4        C6

           H2N        O

            C3        C5
                 C4        C6

           H2N        O


Manuscript in preparation
             G-Quadruplex Formation
                                     Increase in G-quadruplex formation with
•TTGGGGT forms parallel strand
G – quadruplex

•Potassium ion stable

•Presence of anthracene
acrylamide increases G-
quadruplex component

•DOSY studies underway

Zhou, Q.; Lin, L.; Xiang, J.; Sun, H.; Tang, Y.; Fast screening and structural
elucidation of G-quadruplex ligands from a mixture via G-quadruplex recognition and
NMR methods. Biochimie, 2008, 1-5.
•   Prof Nikolai Kuhnert, Jacobs Bremen
•   Dr Jean-Marie Peron, Kingston
•   Judith Peters, Surrey
•   RSC
•   Invitrogen
• It has been approximately 15 years since telomerase
  was described as an almost universal marker for
  human cancer.
• Shortened telomeres undergo replicative senescence
• TRAP Assay inappropriate for G-Quadruplex ligand
  activity measurement Paper – De Clan et al. PNAS,
• Grant applications currently pending,
   – Cancer Research UK
   – Royal Society
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•   Cristofari, G.; Lingner, J.; EMBO Journal. 2006, 25, 565-574.
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    2009, 7, 76-84.
•   b) Kuhnert, N.; Le-Gresley, A.; Organic and Biomolecular Chemistry, 2005, 3, 11, 2175-2182.
•   Lehn, J-M.;Chemistry. A European Journal, 1999, 5, 2455-2463.
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•   a) Bugaut, A.; Jantos, K.; Wietor, J.L.; Rodriguez, R.; Sanders, J.K.M; Balashubramamnian, S.; Angew. Chem. Int. Ed. 2008, 47, 2677-2680.
•   b) Kuhnert, N.; Le-Gresley, A.; Tetrahedron Letters. 2008, 49, 7, 1274-1276.
•   Wunz, T.; Dorr, R.; Alberts, D.; Tunget, C.; Einspahr, J.; Milton, S.; Remers, W.; Journal of Medicinal Chemistry, 1987, 30, 8, 1313-1321.
•   Shi, B and Greaney, M.; Chemical Communications, 2005, 886-888.
•   Naasani, I.; Oh-hashi, F.; Oh-hara, T.; Feng, W. Y.; Johnston, J.; Chan, K.; and Tsuruo, T.; Cancer research. 2003, 69, 824-830.
•   (a) Whitney, A.; Ladame, S.; Balasubramanian, S.; Angew. Chem. Intl. Ed. 2004, 43, 1143-1146.
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