Assignment 6

Drug

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Drug+ targeting carrier

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Questions
1.

Drug development process of a drug carrier currently on the market. Preclinical studies for a drug targeting carrier- protein Formulation studies for a drug targeting carrier-protein for oral use

2.

3.

4.

Biomarkers to show the effectiveness of a drug targeting carrier with the anti-fibrotic 4/21/12 compound in animals and man

Drug Doxil: liposomal form of doxorubicin development

bind DNA and inhibit nucleic acid synthesis
v

The active ingredient : doxorubicin HCl

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Targ Liposomes show preferential homing into tissues with a et sinusoidal capillary system

Liposomes offer the opportunity of targeting large amounts of drug to liver, spleen and lung after i.v. injection

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Lead
Liposomes
microscopic vesicles composed of a phospholipid bilayer . drugs is in a nanocage-functionalized with channel proteins. the drug molecules diffuse through the channel, driven by concentration difference.

The STEALTH liposomes of DOXIL

Formulation Surface-bound methoxypolyethylene glycol (MPEG): protect liposomes from detection by the mononuclear phagocyte system (MPS) increase blood circulation time. penetrate the altered vasculature of tumors
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Pharmacokin Single and repeated intravenous doses in rats and dogs etics encapsulated in Intravenous administration of doxorubicin
Distribution/toxicity: heart, bone marrow and kidneys Experimental set up: localize doxorubicin in the tissues of nude mice bearing human prostatic carcinoma xenografts

increases AUC, T1/2 reduced clearance increased therapeutic index for Doxil.

PEG liposomes

v The long circulation time of Doxil results in higher drug levels and longer persistence of drug in solid tumors.
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Effica cy

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The altered pharmacokinetics and tissue disposition Improved the efficacy of the drug

Acute and repeat dose studies in several species cardiotoxicity, nephrotoxicity, myelotoxicity and dermal toxicity. Liposomes carcinogenesis and mutagenesis toxicity: In vitro (mouse lymphoma and chromosomal aberration assays) In vivo (mammalian micronucleus assay) Impairment of fertility: In vivo (mice, rats, dogs) Ovarian and testicular atrophy Hypospermia Degeneration of seminiferouse tubules Reproductive toxicology: embryotoxicity

Toxicit y

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SAFET Y!
Formulation Long term toxicology studies PK Side-effects: cardiac toxicity (11%), infusion reaction (7.1%), myelosuppression (40.2%), HFS (50.6%) Toxicity Potentiation New regime of administration

Max tolerat ed dose 4/21/12

Liver cirrhosis : irreversible

inflammatory disease liver break down loss of function

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Preclinical studies for drug targeting carrier- protein

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Carrier Suc-HSA Aco-HAS Form-HSA Man10-HSA

SEC +++ +++ ++ +

KC ++ +++ +++ + +

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Drug Targeting Organ-Specific Strategies. Edited by G. Molema, D. K. F. Meijer

Mal-BSA ND Nap20-HAS +++ Dexa10HSA +++

4.

Biodistribution

(radiolabelling, measured in homogenates of the In organs) vivo!

In vitro

(animals and human tissues).

4.

Intrahepatic distribution
Immunohistochemistry of perfused liver lobes or liver slices

5.

Uptake mechanism
in vitro: cell culture, tissue, slices

6.

Release study Efficacy

In vivo (BDL, BDO and tetrachloride-induced models ) : 4/21/12

7.

9.

PK (PK-PD) modeling (efficacy/toxicity)

In vitro PK modeling: model of Hunt, model of Stella and Himmelstein, model of Boddy

TA =Csstarget(DC) / Csstarget(D) DTI =AUCtarget(DC)*AUCtox(DC) / AUCtarget(D)*AUCtox(D) TI = mTD / mED

TA > 1 - drug has been targeted successfully

DrugPD study :Organ-Specific Strategies. Targeting single and repeat dose 4/21/12 Edited by G. Molema, D. K. F. Meijer

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Formulation Studies

The goal of an ideal drug delivery system is to deliver a drug to a specific site, in specific time and release pattern Pass toxicity studies

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Formulation

Administration, absorption, target finding, clearance

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Protein as a carrier

Administration

orally tablet or capsule high proteolytic activity in the Gastrointestinal tract Solution: Enteric coating release in the ilieum

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Protein as a carrier

Absorption

Over epithelial cell in ilieum Poor penetration intestinal membrane due to:

Molecular size and hydrophilic characteristic

Rule of Five

Log P between 1-5; mass <500Da; H-donor <5; Hacceptors (N/O atoms) <10

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Protein as a carrier

Solutions:

Increase membrane permeability Modify protein for receptor recognition by endogenous cellular transport systems (e.g. glucose
transporter, bile-azid transporter, vitB12 rec., Fc rec.)

Increase lipophilicity with functional groups

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Distribution and target finding

Portal vein to liver Target recognizing domain Drug release from protein by cleaving

Clearance

Carrier degradation for kidney clearance

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Protein Carrier Properties

High bioavailability Constant efficacy Biodegradability Lack of toxicity Soluble properties Robustness Quality Cheap

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Enteric Coating Formulation

What is enteric coating? Coating Material and Process

methyl

acrylate-methacrylic acid copolymers acetate succinate propyl methyl cellulose phthalate

cellulose hydroxy fast etc. 4/21/12

release in the duodenum

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Absorption in the intestine

Disintegration from the dosage form absorption
Optimal absorbtion:
v

absorption in the duodenum, where the protein is pH stable

v

Comply with Rule of Five Log P between 1-5; mass <500Da; H-donor <5; Hacceptors (N/O atoms) <10

If protein size is large

active absorption

(must be negatively charged!)

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v

Drug Release

Specific linkage between drug and carrier Protein must be degradable

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Biomarkers

Biological processes, pathogenic processes, pharmacologic responses Liver cirrhosis --> fibrogenesis. Anti-fibrotic drug carrier In vitro studies, animal models, in vivo human studies

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In vitro studies

Culture Activated HSC and Hepatic Stellate Cell Lines Culture of Other Cell Types Implicated in Hepatic Fibrogenesis Precision-cut Tissue Culture Slices
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Biomarkers in animals

Rodent models

mouse preferred!

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Class 1 and class 2 fibrotic biomarkers

Class 1

Serum, urine, biopsy Extracellular matrix turnover Changes fibrogenic cell types (Hepatic Stellate cells and fibroblasts) Relate to activity fibrogenic process, not stage of fibrotic transition. Expensive Whats going on

Class 2

Biochemical scores and biomarker panels

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Degree of fibrosis Not fibrogenic cell changes, no

Current state

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Problem
The non-invasive classical biomarkers are not sufficient enough. New non-invasive single biomarkers are promising

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Novel biomarkers

Tropomyocin MFAP-4

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Thank you for attention!

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