Novel Drug Design Modified Megestrol by Group II

Introduction Hepatocellular carcinoma  Occur almost in patients witn - Chronic hapatitis virus C and/or B infection - Cirrhosis  Represent the final step of the natural course for virus induced liver disease  More than 500,000 people are diagnosed each year throughout the world and over a million death per year  More common in developing country in Africa and East asia

 More frequent in men than in women  No specific drug for the treatment  Risk factors: - HVB - HVC - aflatoxin - alcohol - sex hormones Geographic distribution of hepatocellular carcinoma.Incidence rates (%) in total population A, female; B, male.

Estrogen Receptor (ER)  Receptor for estrogen located intracellular in many organs  Contain a specific site to which only estrogens (or closely related molecules) can bind  Act as a transcription factor, regulate the reading of DNA and production of protein  Two different ER are usually call  and  receptor

Estrogen function as signaling molecule

Estrogen Receptor in Liver  ER  has been well characterized in human liver Normal Liver  wild-type ERs Hepatocellular carcinoma  wild-type ERs  Variant form of ER (vER) exon 5 deletion (ER  5)

Variant from of Estrogen Receptor and Hepatocellular carcinoma  vER largely predominates in HCC  vER appears most frequent in patients infected with Hepatitis B virus  Growth rate of HCC in patient with vER higher than patient with wtER  vER  elevate proliferation rate  tumor aggressiveness  lack of hormonal control on tumor growth  (ER  5) ---- > lack the hormone binding domain but being intact in the DNA-binding domain

Chemotherapy “The use of chemical substances to treat the disease” Types Alkylating agents Plant Alkaloids Antitumor Antibiotics Antimetabolites Topoisomerase inhibitors Miscellaneous Antineoplastics Hormonal therapy

Alkylating agents Add alkyl groups to many electronegative groups e.g Nitrogen mustard (cytotoxic chemotherapy) Hormonal therapy e.g.  Tamoxifen  Megestrol acetate Competitive binding to the receptor and block the action of hormone and thereby interfere with, or even prevent, the proliferation of cancer cell

Megestrol acetate  A synthetic female hormone belonging to the progesterone group  Survival of HCC patients therapy with megestrol acetate is increase  Slowdown tumor growth  Drug able to block both wtER and vER  Anti estrogen action  Usually for women whose cancers do not respond to the other hormone treatments

Bifunctional molecule  Produce DNA adducts  Specific bind the estrogen receptor - Inhibit DNA repair - Induction of growth inhibition, apoptosis and antitumor activity - Consist of => War head => Linker => ligand binding domain Modified Megestrol

Presentation Outline - Production Ms. Jittima Khorungkul - Mechanism Testing of the Drug Mr. Pasavi Ratchapongsirikul - Preclinical Study Ms. Sirikan Nawapan - Clinical Trial Ms. Carolina Rusdy Akib - Marketing Mr. Mahinda Chandrasiri Edirisooriya

Production of Modified Megestrol by Jittima Khorungkul

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

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

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

Adduct shielded from Repair DNA repair enzyme Adduct persists

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

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

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

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

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

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

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

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

Synthesis Procedure

Final Product: Modified megestrol

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

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