Catalysis of substitution reactions on heterocycles bound to a solid phase Seminar in honor of Prof. Marv Caruthers, Boulder, CO, USA, 2001

1. Catalysis of Substitution Reactions on Heterocycles Bound to a Solid Phase [email_address]

2. Catalysis of Substitution Reactions on Heterocycles Bound to a Solid Phase

3. Levoglucosan based Libraries epoxide opening epoxide opening difficult alkylation or acylation avoid glycosidic scission O O X O O O N H 2 R 1 R 2 R 3 Pd mediated coupling Pd mediated coupling

6. Schematic Representation of Resins crosslink shrunken resin (poorly solvated) compound bound to the polymer Reactions can only be performed, if reagents can diffuse through the gelphase of the beads. The polymer has to be solvated with the appropriate solvent to form such a phase. swollen resin (solvated)

13. Purines may bind to Proteins in different ways They address nucleoside binding pockets

14. Nucleoside Cofactors Lead us to Drug Targets 1 . Many cofactors contain nucleoside motifs 2. Nucleosides or nucleoside cofactors are involved in all important cellular processes. DNA synthesis RNA synthesis protein synthesis carbohydrate synthesis and oligomerization lipid synthesis and processing synthesis of homo and heterocyclic aromatics signaling via phosphoylation and sulfatation signaling as second messenger or hormone alkylation and dealkylation of DNA and other substrates. 3. Nucleoside binding sites are not optimized toward very tight binding!

15. i, ii: Attachment to the polymer iii: 6-Substitution iv: 2-Substitution v: Bromination vi: Stille coupling vii: Cleavage Reaction Scheme

16. Immobilization of 2,6-dichloropurine

17. C6-Substitution

18. C6-Substitution Reaction temperature has to be 53°C to avoid substitution on C2 for very nucleophilic amines (N-CH 2 CH 2 NH 2 , C-CH 2 CH 2 NH 2 piperazines, piperidines). No C2-substitution even at 70°C: anilines, benzylamines, morpholin, primary amines with higher order of substitution on their Ca

19. Sustitution on C2 * rest is starting mat.

21. Two Types of Bromine Complexes Were Investigated Synthesis: Reactivity:

22. Modification of the Polymeric Support by the Brominating agent Bromine-complex Br-content of resin, treated with the Bromine Complex (EA)

23. 5% conversion no reaction: complete conversion: side reactions: not tolerated groups: activated aromatics: bromination amines: many oxidation products Reactivity of the Bromine Complex with Purines: Solvent: NMP R 1 : H, alkyl, R 1 ’: alkyl R 2 : H, alkyl, R 2 ’: H, alkyl tolerated groups: CONR 2 , CONRH CONH 2 , ether functions, aromatics more deactivated than Bn

25. The Reactivity of the Bromine Complex in wet Benzene: Side chain oxidations are not mediaded by the solid phase. They work are observed also in solution phase .

26. Proposed Mechanism for the Modification at C2 i: Br 2 -complex; ii: H 2 O

27. Dehalogenation during Suzuki couling conditions ArB(OH) 2 Pd-cat Attempted Synthesis to Yield 2,6,8-Trisubstituted Purines

28. Substitution on C8 dppp:

29. Selection of Buildingblocks: 1) Size and shape constraints: selection of privileged structures based on modeling previous screening results, docking excercises 2) Availability of building blocks: emphasis on proprietary building blocks, commercial building blocks 3) Chemtox considerations: exclusion of: -NO 2 ,-NO,-N 2 + , I, -I=O, -N 3 , heavy metals, alkylating agents, acylating agents, hydrazines, SH, aniline-functions in product 4) Tests of building blocks (several hundred reactions) 5) Generation of a virtual library with the building blocks which work: Agreement to rule of 5 checked, exclusion of dramatic outliers 6) Final selection for Synthesis

30. The Synthesis is performed on PS-Beads PS-beads 60 mg per Kan Rf -transponders with unique ID Read only ! Reading interval: 0.1s RF-frequency: 125 KHz Reading distance : 1 cm reusable many times ! 1 per Kan Now all Kans can be distinguished 1 cm

34. SynthMan Plan synthesis Perform synthesis Purify & Analyze compounds Archive data & stock HPLC Weighing full Dilution Import BB MS Import Compounds Family data Reagent data Method data Salt data Barcodes Working Sheet Preparereagents Component Database Family Database HPLC (Millennium) Structural Analysis Software for Controlling of the Production, Purification and Analyzing Process Weighing empty LC-MS (MicroMass) prep. LC Automatic Sampler (Hamilton) 2. 1. 9 16. 8. 7. 15. 6. 5. 3. 4. 14. 13. 12. 10. Archive Databases Archive Databases Automatic weighing (MicroWeight) Automated Sorting (Kan Sort) Sort to flasks and racks 11.

35. Immobilization of 2,6-Dichloropurine

36. Addition of Resin slurry to IRORI MiniKans

37. Sort & Combine transponder Sort Kans redistribute

38. Washing of Kans TM in Bulk By R. Gallarini, S. Vinzenz,W. Brill

39. Sorting of Kans Into Racks prior to Cleavage: Last step of Sort-and-Combine Method

40. TFA Transponder TFA-Cleavage

42. MicroMass LC-MS HPLC-MS correlation of fractions with expected mass fraction location LC-fractions crude reaction mixtures location of crude reaction mixtures, expected mass

43. High-throughput purification Crude sample Fraction collection by LC/MS Result of purification Confirmation of identity HPLC/UV purity : 20% HPLC/UV purity : >95%

44. Evaporation

45. Automated weighing system Throughput: 1 rack / hr

46. Recycling of used Kans By S. Vinzenz,W.Brill

47. Batch-Registration of Compounds by Synthman

48. Achnowledgement Modeling: E. Jacoby Synthman: R. Fäh, H.-P. Moessner Synthesis: S. Müller, J. Schaub, C. Riva-Toniolo*, D. Tirefort ** Purification & Registration: F. Gombert, G. Lerch, H. Wettstein Hardware: S. Vinzenz, R. Gallarini General: A. DeMesmaeker, J. Zimmermann, S. Wendeborn