Jessica Scott May 17, 1999

Phospholipids
Polar Head Groups

Three carbon glycerol

What is a liposome?
Spherical vesicles with a phospholipid bilayer
Hydrophilic

Hydrophobic

Cell Membrane

Uses of Liposomes
Chelation therapy for treatment of heavy metal poisoning Enzyme replacement Diagnostic imaging of tumors Cosmetics Study of membranes

Drug Delivery

Why Use Liposomes in Drug Delivery?

Drug Targeting
Inactive: Unmodified liposomes gather in specific tissue
reticuloendothelial system Active: alter liposome surface with ligand (antibodies, enzymes, protein A, sugars)

Physical: temperature or pH sensitive liposomes Directly to site

Why Use Liposomes in Drug Delivery?

Pharmokinetics - efficacy and toxicity
Changes the absorbance and biodistribution Deliver drug in desired form Multidrug resistance

Protection
Decrease harmful side effects
Change where drug accumulates in the body

Protects drug

Why Use Liposomes in Drug Delivery?

Release
Affect the time in which the drug is released

Prolong time -increase duration of action and decrease administration Dependent on drug and liposome properties
Liposome composition, pH and osmotic gradient, and environment

Modes of Liposome/Cell Interaction

Adsorption Endocytosis

Fusion

Lipid transfer

Classes of Liposomes
Conventional Long circulating

Immuno

Cationic

Liposomes Help Improve

Therapeutic index Rapid metabolism Unfavorable pharmokinetics Low solubility Lack of stability Irritation

Custom design

Lipid content Size Surface charge Method of preparation

Current liposomal drug preparations

Type of Agents Anticancer Drugs Anti bacterial Examples Duanorubicin, Doxorubicin*, Epirubicin Methotrexate, Cisplatin*, Cytarabin Triclosan, Clindamycin hydrochloride, Ampicillin, peperacillin, rifamicin AZT cDNA - CFTR* Hexosaminidase A Glucocerebrosidase, Peroxidase In-111*, Tc-99m Amphotericin B* Malaria merozoite, Malaria sporozoite Hepatitis B antigen, Rabies virus glycoprotein

Antiviral DNA material Enzymes Radionuclide Fungicides Vaccines

*Currently in Clinical Trials or Approved for Clinical Use

CFTR
Gene Therapy
Deliver cDNA of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) to epithelial tissue of respiratory system

Cationic liposome
Fuse to cell membrane and incorporate cDNA into cell

Clinical trials - no significant change in symptoms

Now trying adeno associated virus

Doxil
Chemotherapy drug doxorubin
Anemia, damage to veins and tissue at injection, decrease platelet and WBC count, toxic to

Treats Kaposis sarcoma lesions or cancer tumors Modifications of liposome stealth

keeps doxorubin in blood for 50 hours instead of 20 minutes concentrates at KS lesions and tumors

*Just approved by FDA*

Amphotericin B
Systemic fungal infections in immune compromised patients AmB - kills ergosterol-containing fungal cells, also kills cholesterol-containing human cells Side effects: nephrotoxicity, chills, and fevers Fungizone - AmB with deoxycholate

Liposomal Formulation of AmB

Phospholipid:AmB ratio

AmB

Cholesterol - only few %moles

Lipid

Exact Mechanism of liposomes not understood

Diffusion Lipid transfer

Decrease in toxicity No decrease in effectiveness of drug against fungi

Problems with Liposomal Preparations of Drugs

$$$$
Fungizone $40.58 Amphotec $2334 Doxil $1200 per treatment, twice the cost of normal protocol of chemotherapy and drugs

Lack long term stability (short shelf life)

Physical and chemical instability

Freeze dry and pH adjustment

Low Pay Load - poor encapsulation

Polar drugs and drugs without opposite charge Modifications

Problems continued
Possibility of new side effects
Doxil hand and foot syndrome

Efficacy
CFTR

Future
Studies with insulin show that liposomes may be an effective way to package proteins and peptides for use Clinical Trials for several liposomal formulations More studies on the manipulation of liposomes

Journals
Allen, Theresa M. "Liposomal Drug Formulations: Rationale for Development and What We Can Expect for the Future." Drugs 56: 747-756, 1998. Allen, Theresa M. "Long-circulating (sterically stabilized) liposomes for targeted drug delivery." TiPs 15: 214-219, 1994. Allen, Theresa M. "Opportunities in Drug Delivery." Drugs 54 Suppl. 4: 8-14, 1997 Janknegt, Robert. "Liposomal and Lipid Formulations of Amphotericin B." Clinical Pharmacokinetics. 23(4): 279-291, 1992. Kim, Anna et al. "Pharmacodynamics of insulin in polyethylene glycol-coated liposomes." International Journal of Pharmaceutics. 180: 75-81, 1999. Quilitz, Rod. "Oncology Pharmacotherapy: The Use of Lipid Formulations of Amphotericin B in Cancer Patients." Cancer Control.5:439-449, 1998. Ranade, Vasant V. "Drug Delivery Systems: Site-Specific Drug Delivery Using Liposomes as Carriers." Pharmacology. 29: 685-694, 1989. Storm, Gert and Daan J.A. Crommelin. "Liposomes:quo vadi?" PSTT 1: 19-31, 1998. Taylor, KMG and JM Newton. "Liposomes as a vecicle for drug delivery." British Journal of Hospital Medicine. 51: 55-59, 1994

Websites
James, John S. "Doxil Approved for KS." www.immunet. org.imminet/atn.nsf/page/a-235-03. Wasan, Ellen. "Targeted Gene Transfer." Member.tripod.com/~rrishna/lipos1.html "Introduction to Controlled Drug Delivery Systems." www5.bae.ncsu.edu/bae/reearch/blak k/otherprojects/drugDeliver_97/ http://www. Mssm.edu/medicine/thrombosis/phosphol.html "Doxorubicin." http://tirgan.com/adria.htm "Clinical Pharmacology Online." http://www.cponline.gsm.com/scripts/fullmono/showmono. "Drugstore.com" http://www.drugstore.com/pharmacy/prices/Amphotec. "Sequus' Doxil Becomes First Liposome Product Approved In U.S." www.slip.net/~mcdavis/ database/doxor_1 "Liposomes." www.collabo.com/liposom0.htm
Paustin, Timothy. Cellular Membranes.www.bact.wisc.edu/microtextbook/bacterialstructure/Membranegen.html www.cbc.umn.edu/~mwd/cell_www/chapter2/membrane.html#PHOSPHOLIPIDS

Books
Jones, Macolm N. and Chapman, David. Micelles, Monolayers and Biomembranes. Wiley-Liss. New York (1995). Garrett, R. and Grisham C. Biochemistry, 2nd ed. Saunders Colleges Publishing. New York (1999). 264.