1. Production of Modified Megestrol by Jittima Khorungkul

2. Modified Megestrol Production Goal: Synthesis bifunctional molecule that can use in Liver cancer treatment Bifucntional molecule:  Produce DNA adduct  Specific binding the estrogen receptor with high affinity

3. Megestrol Estrogen receptor DNA adduct Binding to vER Bifunctional molecule structure Ligand Domain Linker War Head

4. Expression of essential gene Undamaged cell Nuclear protein (e.g.ER) promoter How modified megestrol work….. Estrogen and ER complex

5. Adduct shielded from Repair DNA repair enzyme Adduct persists

6. Adduct shielded from Repair In non-cancer cell (less express of vER) DNA repair enzyme adduct

7. Adduct shielded from Repair Cancer cell (over express of vER)

8. Adduct “Hijacks” Transcription Factor X Cancer cell (over express of vER)

9. Modified megestrol N,N -bis-chloroethylaniline (War Head) Alkyl-amino-carbamate (Linker) Megestrol acetate (Ligand Domain) Consisted of : (CH 2 ) 6 -N-(CH 2 ) 2 -0 N-(CH 2 ) 3 - -N H O H (CH 2 ) 2 C l Ligand DomainLinker War Head

10. Modified megestrol Megestrol acetate (Ligand Domain) -Binding to the linker at 7 alpha position  Large alkyl groups can be attached with retention of high affinity for ER Megestrol acetate 7 alpha position

11. Modified megestrol N,N -bis-chloroethylaniline (War Head) - Ability to alkylate DNA - From covalent DNA adduct at the N7 position of guanines

12. Modified megestrol Alkyl-amino-carbamate (Linker) Consist of - amino - carbarmate group  provide a relatively rigid connection  resistant to hydrolytic enzyme

13. Synthesis Procedure (1) (2) (3) (4)(5)

14. Synthesis Procedure

15. Final Product: Modified megestrol

16. Properties of Modified Megestrol Chemical Formula: C 42 H 65 Cl 2 N 3 O 4 Exact Mass: 745.44 Molecular weight: 746.89 Element Analysis: C, 67.54; H, 8.77; Cl, 9.49; N, 5.63; O, 8.57

17. Summary Modified Megestrol -Modified Megestrol is the bifunctional molecule that consist of ‘Warhead’, ‘Linker’ and ‘Ligand binding domain’ -It has the abilities to produce DNA adduct and capable of binding the vER - vER-DNA adduct complexes will shielded from DNA repair enzyme War head Linker Megestrol acetate