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Nanocell: Nanocell: Mechanism of actionMechanism of action

Soraya AroonvilairatSoraya Aroonvilairat

Inter-U program, AITInter-U program, AIT

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Scope: Mechanism of NanocellScope: Mechanism of Nanocell

• MechanismMechanismss of of chemotherapeutic agentschemotherapeutic agents• CConventional drugs for liver canceronventional drugs for liver cancer & & drug resistance drug resistance

problemsproblems• Mechanisms of Mechanisms of FTY720, doxorubicinFTY720, doxorubicin & 5FU& 5FU

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httphttp://://wwwwww..ovcovc..uoguelphuoguelph..caca//BioMedBioMed//CoursesCourses//PublicPublic//PharmacologyPharmacology//pharmsitepharmsite//98-40998-409//CancerCancer//Anticancer_drugs1Anticancer_drugs1..htmlhtml

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Types of chemotherapeutic drugs (1)Types of chemotherapeutic drugs (1)

• AntimetabolitesAntimetabolites– Structurally related to naturally occurring compounds Structurally related to naturally occurring compounds

(vitamins, amino acids, nucleotides)(vitamins, amino acids, nucleotides)

– Result: interfere DNA & RNA synthesis, cell proliferationResult: interfere DNA & RNA synthesis, cell proliferation

• Covalent DNA-binding drugs (Alkylating agents)Covalent DNA-binding drugs (Alkylating agents)– Drugs (electrophilic) covalently bind to alkyl groups of Drugs (electrophilic) covalently bind to alkyl groups of

bases of DNA & protein (nucleophilic)bases of DNA & protein (nucleophilic)

– Result: single strand break, base mispairing, cross linkage Result: single strand break, base mispairing, cross linkage

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Types of chemotherapeutic drugs (2)Types of chemotherapeutic drugs (2)

• Drugs affecting endocrine function Drugs affecting endocrine function – Steroid hormones (estrogen) or their antagonistsSteroid hormones (estrogen) or their antagonists

• Inhibitors of chromatin functionInhibitors of chromatin function– Drugs which disrupt the chromosomal dynamic necessary Drugs which disrupt the chromosomal dynamic necessary

to carry out DNA replication and mitosisto carry out DNA replication and mitosis

– 2 subgroups: topoisomerase inhibitors & microtubule 2 subgroups: topoisomerase inhibitors & microtubule inhibitorsinhibitors

• Noncovalent DNA-binding drugsNoncovalent DNA-binding drugs – Intercalating drugs forming tight drug-DNA interactionIntercalating drugs forming tight drug-DNA interaction

– Free radical damage cause single strand breakFree radical damage cause single strand break

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CisplatinCisplatin

• IUPAC name : IUPAC name : dichloroplatinumdichloroplatinum;azanide;azanide

• Chemical formula : ClChemical formula : Cl22HH44NN22PtPt

• Platinum based Platinum based chemotherapeutic drug & chemotherapeutic drug & Alkylating agentAlkylating agent

• Application: solid tumors (testis, Application: solid tumors (testis, ovary, head & neck, bladder)ovary, head & neck, bladder)

Pt

Cl

H3N Cl

H3N

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Cisplatin-DNA interactionCisplatin-DNA interaction

httphttp://://wwwwww..iupaciupac..orgorg//publicationspublications//pacpac//19871987//pdfpdf//5902x01815902x0181..pdfpdf

NN77-guanine-guanine

Inter-strand Inter-strand cross-linkscross-links

Intra-strand Intra-strand adductadduct

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Mitomycin CMitomycin C

• Chemical formula : CChemical formula : C1515HH1818NN44OO55

• Antitumor antibiotic, alkylating Antitumor antibiotic, alkylating agentagent

• Application: Application: malignant malignant neoplasm (oral cavity, pharynx, neoplasm (oral cavity, pharynx, breast, and urinary bladder)breast, and urinary bladder)

• Mechanism: binding to DNA, Mechanism: binding to DNA, cross-linking, inhibit DNA cross-linking, inhibit DNA synthesissynthesis

N

O

NH2

O

O

NH

O

NH2

O

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What are the problems of What are the problems of chemotherapeutics usage chemotherapeutics usage

• Side effects: nausea & vomiting, hair loss, bone Side effects: nausea & vomiting, hair loss, bone marrow suppression, mouth sores, skin changes, marrow suppression, mouth sores, skin changes, diarrheadiarrhea

• Drug Drug resistance: resistance: – Decrease drug accumulation ( drug influx, drug efflux)Decrease drug accumulation ( drug influx, drug efflux)

– Altered drug metabolismAltered drug metabolism

– Altered drug targetsAltered drug targets

– Increased repair of drug-induced damageIncreased repair of drug-induced damage

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Annu Rev Med 53:615-27, 2002

Mechanisms of drug resistanceMechanisms of drug resistanceABC transporters ABC transporters

(P-gp)(P-gp)

Altered drug Altered drug accumulation within accumulation within

cellscells•Binding of drugsBinding of drugs

•Activate ATP-Activate ATP-binding domainbinding domain

•ATP hydrolysis ATP hydrolysis causes drug release causes drug release into extracellular into extracellular spacespace

•Mutation of receptors Mutation of receptors or transporters involving or transporters involving drug uptakedrug uptake

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200 nm

100 nm

Nanocell design for liver cancerNanocell design for liver cancer

NanocoreNanocore

Loaded with ChemotherapeuticsLoaded with Chemotherapeutics

Slow release kineticsSlow release kinetics

Lipid vesicle with integrated Lipid vesicle with integrated antiangiogenicsantiangiogenicsFast release kineticsFast release kinetics

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Anti-angiogenic: FTY720Anti-angiogenic: FTY720

• IUPAC name: IUPAC name: 2-amino-2-[2-(4-octylphenyl)ethyl] propane-2-amino-2-[2-(4-octylphenyl)ethyl] propane-1,3 diol1,3 diol

• Synthetic analogue of natural compound derived from Synthetic analogue of natural compound derived from fungus fungus Isaria sinclairii.Isaria sinclairii.

• Originally developed as novel immunosuppressant in organ Originally developed as novel immunosuppressant in organ transplantation transplantation

• Sphingosine analogueSphingosine analogue

OH

NH2

OH

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FTY720: sphingosine analogueFTY720: sphingosine analogue

httphttp://://wwwwww..postechpostech..acac..krkr//chemchem//skcskc--lablab//researchresearch//lipidomicslipidomics..htmhtm

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Angiogenic processAngiogenic process

www.amplab.de/3D-Images/angiogenesis-scheme.gifwww.amplab.de/3D-Images/angiogenesis-scheme.gif

•Angiogenesis induced by VEGF (tumor)

•Degradation of basement membrane

•Invasion & migration of endothelial cells to tumor

•Elongation of new vessel by proliferation

•Vessel maturation by synthesis of ECM

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• FTY720 phosphate (FTY-P) acts aFTY720 phosphate (FTY-P) acts a high-affinity high-affinity agonist at the G protein-coupled sphingosine 1-agonist at the G protein-coupled sphingosine 1-phosphate receptor phosphate receptor

• DecreasedDecreased Rac expression of tumor cells (related to Rac expression of tumor cells (related to endothelial cells migration, chemotaxis) endothelial cells migration, chemotaxis)

• Inhibited VEGF expression induced vascular Inhibited VEGF expression induced vascular permeability (anti-angiogenesis)permeability (anti-angiogenesis)

FTY720: mechanism of actionFTY720: mechanism of action

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FTY720: mechanism of actionFTY720: mechanism of action

httphttp://://wwwwww..postechpostech..acac..krkr//chemchem//skcskc--lablab//researchresearch//lipidomicslipidomics..htmhtm

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Doxorubicin (Adriamycin)Doxorubicin (Adriamycin)

• Chemical formula : Chemical formula : CC2727HH2929NONO1111

• Cytotoxic anthracycline Cytotoxic anthracycline antibioticantibiotic

• Noncovalent DNA binding Noncovalent DNA binding drugdrug

• Application: treatment of solid Application: treatment of solid tumors (breast, ovarian, tumors (breast, ovarian, lymphoma, HCC, soft tissue lymphoma, HCC, soft tissue sarcoma)sarcoma)

O

O

OH

OH O

OH

O

OH

O

O

OH

NH2

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Doxorubicin: mechanism of action Doxorubicin: mechanism of action

• DNA intercalation: bDNA intercalation: bindindinging to DNA to DNA, , inhibitinhibit the the progression of topoisomerase II which unwinds DNA progression of topoisomerase II which unwinds DNA for transcription,for transcription, consequent in cell cycle disruption consequent in cell cycle disruption & cell death& cell death

• Free radical damageFree radical damage involving reactive oxygen involving reactive oxygen species (ROS) ss breakagespecies (ROS) ss breakage

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Doxorubicin-DNA complexDoxorubicin-DNA complex

httphttp://://chemistrychemistry..clemsonclemson..eduedu//ChemDocsChemDocs//

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5-Fluorouracil (5-FU)5-Fluorouracil (5-FU)

• IUPAC name : IUPAC name : 5-fluoro-1H-pyri5-fluoro-1H-pyrimidine-2,4-dione midine-2,4-dione

• Chemical formula : CChemical formula : C44HH33FNFN22OO22

• Anti-metabolite drugAnti-metabolite drug• Pyrimidine base analoguePyrimidine base analogue• Application: treatment of solid Application: treatment of solid

tumors (breast & colon tumors (breast & colon carcinoma)carcinoma)

NH

NH

O

F

O

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5-FU: pyrimidine analogue5-FU: pyrimidine analogue

• 5-FU resembles the pyrimidine bases uracil and 5-FU resembles the pyrimidine bases uracil and thymine (components of RNA & DNA)thymine (components of RNA & DNA)

NH

NH

O

F

O

NH

NH

O

CH3

O

NH

NH

O

O

Uracil Thymine 5-FluorouracilUracil Thymine 5-Fluorouracil

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5-FU: mechanism of action5-FU: mechanism of action5-FU5-FU

FdUMPFdUMP FUTPFUTP

Inhibit thymidylate Inhibit thymidylate synthasesynthase

ApoptosisApoptosis

Mistakenly Mistakenly incorporate to RNAincorporate to RNA

alter RNA processing alter RNA processing and protein synthesisand protein synthesis

Depletion of dTMP Depletion of dTMP (required for DNA systhesis)(required for DNA systhesis)

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SummarySummary

• Nanocell is designed to solve the problems of side Nanocell is designed to solve the problems of side effects & drug resistanceeffects & drug resistance

• Design of nanocell: anti-angiogenics (outer), Design of nanocell: anti-angiogenics (outer), chemotherapeutics (inner)chemotherapeutics (inner)

• Anti-angiogenesis of FTY720: via Rac-VEGF Anti-angiogenesis of FTY720: via Rac-VEGF mediated pathwaymediated pathway

• Chemotherapy of Doxorubicin: DNA intercalation, Chemotherapy of Doxorubicin: DNA intercalation, 5FU:apoptosis, alter RNA processing & protein 5FU:apoptosis, alter RNA processing & protein synthesis synthesis