EP0525154A1 - Interaction entre des liposomes bioadhesifs et des sites cibles - Google Patents

Interaction entre des liposomes bioadhesifs et des sites cibles

Info

Publication number
EP0525154A1
EP0525154A1 EP19920904871 EP92904871A EP0525154A1 EP 0525154 A1 EP0525154 A1 EP 0525154A1 EP 19920904871 EP19920904871 EP 19920904871 EP 92904871 A EP92904871 A EP 92904871A EP 0525154 A1 EP0525154 A1 EP 0525154A1
Authority
EP
European Patent Office
Prior art keywords
liposome
component
administration
recognizing substance
liposomes
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP19920904871
Other languages
German (de)
English (en)
Inventor
Rimona Margalit
Theodore Roseman
Ray W. Wood
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Baxter International Inc
Original Assignee
Baxter International Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Baxter International Inc filed Critical Baxter International Inc
Publication of EP0525154A1 publication Critical patent/EP0525154A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers

Definitions

  • the present invention relates to a novel drug delivery system, particularly to microscopic drug delivery systems (MDDS) utilizing drug-encapsulating "bioadhesive" liposomes for topical and local drug administration.
  • MDDS microscopic drug delivery systems
  • topical and local administration of a drug can be in Its free form, dissolved or dispersed in a suitable diluent, or in a vehicle such as a cream, gel or ointment.
  • Examples of therapeutic or designated targets for topical or local drug administration include burns; wounds; bone injuries; ocular, skin, intranasal and buccal infections; ocular chronic situations such as glaucoma; and topically and locally accessed tumors.
  • MDDS Microscopic drug delivery systems
  • particulate systems such as cells, microspheres, viral envelopes and liposomes
  • nonparticulate systems which are macromolecules such as proteins or synthetic polymers.
  • drug-loaded MDDS can perform as sustained or controlled release drug depots.
  • MDDS reduces drug degradation or inactivation.
  • MDDS improves drug efficacy and allows reduction in the frequency of dosing. Since the pharmacokinetics of free drug release from depots of MDDS are different than from directly-administered drug, MDDS provides an additional measure to reduce toxicity and undesirable side effects.
  • Liposomes offer a range of advantages relative to other MDDS systems. Liposomes are lipid vesicles composed of membrane-like lipid layers surrounding aqueous compartments. Composed of naturally-occurring materials which are
  • liposomes are used to encapsulate biologically active materials for a variety of purposes. Having a variety of layers, sizes, surface charges and compositions, numerous procedures for liposomal preparation and for drug encapsulation within them have been developed, some of which have been scaled up to industrial levels.
  • Liposomes can accommodate lipid-soluble drugs, aqueous soluble drugs and drugs with both hydrophilic and hydrophobic residues.
  • Liposomes can be designed to act as sustained release drug depots and, in certain applications, aid drug access across cell membranes. Their ability to protect encapsulated drugs and other characteristics make liposomes a popular choice in developing MDDS, with respect to the previous practices of free drug administration.
  • liposomes as MDDS have limited targeting abilities, limited retention and stability in circulation, potential toxicity upon chronic administration and inability to
  • antibodies can be patient specific and therefore, add cost to the drug therapy.
  • cell-associated entitles can participate in the binding between cells and recognizing substances. These are generally divided into three major types: receptors and non-receptor components of the cellular system and
  • Receptors can be present in several species or states, differing in populations per cell and in binding affinity. Binding to such receptor entitles is usually referred to as "specific binding”. Non-receptor cell membrane components also differ in populations and in affinity. Binding to such non-receptor entitles is usually referred to as
  • administration of drug-encapsulating liposomes should have specificity for and the ability to adhere to the designated target area and should facilitate drug access to intracellular sites.
  • liposomes and other MDDS systems do not meet these performance requirements of topical and local drug administration.
  • recognizing substances are molecules which can be utilized as an adhesive or glue, attaching a drug-encapsulating liposome onto a therapeutic target site.
  • bioadhesive recognizing substances can perform either through receptor mechanisms or through associations with components within the extracellular matrix. Regardless of the specific mechanism of adhesion, these substances are referred to as “bioadhesive recognizing substances” based on their common end result.
  • bioadhesive recognizing substances become an integral part of the liposome, yet remain accessible to the interaction counterpart at the target site. They endow the liposome and encapsulated drug with the ability to adhere to the target site.
  • bioadhesive liposomes have been developed which are target adherent, sustained release drug depots. The identification of recognizing substances and the methodologies of modifying liposomes has been disclosed in concurrently filed applications. These bioadhesive liposomes offer several advantages over previous practices of topically or locally administered free drug and other MDDS, whether with regular liposomes or other MDDS systems.
  • FIG. 1 shows the binding of bioadhesive liposomes
  • Bound liposomes denoted as B, are in units of ng EGF per 10 6 cells.
  • Free ligand concentration denoted as L, are in units of ng EGF per 10 6 cells for bioadhesive liposome (first row of L values) and in units of umoles lipid per 10 6 cells for the regular liposomes (second row of L values).
  • FIG. 2 shows a time course of the binding of bioadhesive liposomes (collagen-modified) of the MLV type to A431 cells in culture (in monolayers). Collagen is tritium-labeled. The fraction of liposomes relative to the amount present in the initial reaction mixture at zero-time which is cell-associated is determined over time.
  • FIG. 3 shows the binding of bioadhesive liposomes
  • Collagen-modified and regular liposomes of the MLV type to A431 cells in culture (in monolayers). Collagen is
  • Bound liposomes denoted as B, are 1n units of 3 -H DPM per 10 5 cells (left scale) and in units of 14 -C DPM per 10 5 cells (right scale).
  • Free ligand concentration denoted as L, are in units of 3 -H or 14 -C DPM per 10 5 cells.
  • Bloadheslve liposome with collagen labeled is depicted with open double triangles;
  • bioadhesive liposome with the liposome labeled is depicted with crosses; and, regular liposome is depicted with asterisks.
  • bioadhesive liposomes have bound to cell cultures having receptors or extracellular matrix which accommodate the recognizing substance bonded to the liposome.
  • Liposomes in particular, multilamellar vesicles (MLV), microemulsified liposomes (MEL) or large unilamellar vesicles (LUVET), each containing phosphatidylethanolamine (PE), have been prepared by established procedures.
  • MLV multilamellar vesicles
  • MEL microemulsified liposomes
  • LVET large unilamellar vesicles
  • PE phosphatidylethanolamine
  • Recognizing substances include epidermal growth factor (EGF), hyaluronic acid (HA), gelatin and collagen.
  • EGF epidermal growth factor
  • HA hyaluronic acid
  • gelatin gelatin
  • collagen a biological origin and are biodegradable and biocompatible.
  • these recognizing substances have functional residues which can be utilized in covalent anchoring to the regular liposomal surfaces.
  • n is the number of different cell-associated binding entities that a cellular system has for a specific recognizing substance
  • [L] is the concentration of free ligand, which can be recognizing substance, free liposomes or bioadhesive liposomes
  • B is the total quantity of bound recognizing substance per given number of cells, at a given [L]
  • Bmax i and Kd i are the total number of sites of a given entity and the corresponding equilibrium dissociation constant.
  • B and B max are normalized for the same number of cells.
  • equation 1 can take the form: where the last term, K ns [L], is the contribution of the non-specific binding to B and K ns is the ratio of Bmax to Kd corresponding to the non-specific binding.
  • A431 cells have been shown to have three classes of EGF receptors, differing 1n their affinities and populations.
  • the first of these classes is the ultra-high affinity sites with an equilibrium dissociation constant of 0.07 nM and a population of 150-4000 sites per cell.
  • the next class is the high affinity sites with an equilibrium dissociation constant of 0.7 nM and a population of 1.5 ⁇ 105 sites per cell.
  • the final class is the low affinity sites with an equilibrium
  • A431 cell cultures were grown in monolayers, in flasks, applying usual procedures for this cell line. Two to three days prior to an experiment, the cells were seeded into multiwell culture plates and the experiments were done when the systems were confluent.
  • the EGF-recogniz1ng substance was labeled with a generally known radioactive marker.
  • Preparation of EGF-modified LUVET was completed as disclosed in the concurrently filed applications.
  • EGF-modified liposomes, free liposomes or free EGF, media was removed from the A431 cells and the cells were washed with a binding buffer.
  • the reaction mixture and cells were incubated for 1-2 hours, at room temperature. Upon dilution and withdrawal of the reaction mixture at the end of incubation, 2-3 successive washings with a binding buffer of the wells were completed. Lysis of cells or detachment of cells from the wells was then followed by withdrawal and collection of the well content, denoted as the cell fraction. Assays of the cell fraction were completed by label counting of the fraction as compared with the counting of the immediate products created through the preparation process.
  • EGF-modified liposome is considerably larger than and different from free EGF, which is expected to affect the binding parameters.
  • the magnitudes of the dissociation constants for EGF-modified liposome systems are expected to be similar to or higher than those of free EGF.
  • the number of receptors per cell that are available for the EGF-modified liposomes is expected to be equal to or lower than the number of available for free EGF. Based on these considerations, the binding data of the present example fit with the receptor classes of ultra-high and high affinities.
  • EGF-modified liposomes bind to this cellular system with high affinity and with a sufficient number of sites for these modified liposomes to perform as the desired bioadhesive liposomes.
  • Binding collagen-modified liposomes to A431 cells was carried out essentially according to the procedures detailed above.
  • the A431 cell line is not known to contain receptors for collagen.
  • the interaction of either free collagen or liposomally bound collagen with the A431 cell line is expected to result from association of collagen with components within the extracellular matrix. Referring to Figure 2, incubation periods up to 4 hours were completed with 3 hours being the optimal period for binding and collagen-liposome concentrations.
  • the binding entitles are of the extracellular matrix type of
  • the dissociation constant for collagen-modified liposomes is expected to be similar to or higher than those of free collagen.
  • the number of available sites in the extracellular matrix available for collagen-modified liposomes is expected to be similar to or lower than free collagen.
  • the example given in Table 2 fits with these considerations.
  • the data for free collagen demonstrate that binding of this bioadhesive recognizing substance to this cellular system does occur and is a measurable phenomena, which can be processed to yield quantitative and meaningful parameters.
  • the data in Table 2 show quite clearly that the binding of
  • collagen-modified liposomes to this cellular system is of sufficiently high affinity and with a large enough number of sites, for the collagen-modified liposomes to perform as the desired bioadhesIve liposomes.

Abstract

Des substances de reconnaissance, le facteur de croissance épidermique, la gélatine, le collagène et l'acide hyaluronique, ont été liés de manière covalente à des surfaces liposomiques et utilisés pour fixer les liposomes sur un site cellulaire cible. L'utilisation de ces liposomes "bioadhésifs" présente plusieurs avantages dans le domaine de médicaments libres à administration locale ou topique. Ces avantages comprennent la protection réciproque à la fois de l'environnement biologique et du médicament, une amélioration de la biodisponibilité et de la rétention du médicament au niveau du site cible; et une adhérence améliorée au site cible désigné.
EP19920904871 1991-02-14 1991-10-30 Interaction entre des liposomes bioadhesifs et des sites cibles Withdrawn EP0525154A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US65501391A 1991-02-14 1991-02-14
US655013 1991-02-14

Publications (1)

Publication Number Publication Date
EP0525154A1 true EP0525154A1 (fr) 1993-02-03

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
EP19920904871 Withdrawn EP0525154A1 (fr) 1991-02-14 1991-10-30 Interaction entre des liposomes bioadhesifs et des sites cibles

Country Status (5)

Country Link
EP (1) EP0525154A1 (fr)
JP (1) JPH05506040A (fr)
AU (1) AU656173B2 (fr)
CA (1) CA2079450A1 (fr)
WO (1) WO1992014446A1 (fr)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9208339D0 (en) * 1992-04-15 1992-06-03 Unilever Plc Treatment composition
WO1994018955A1 (fr) * 1993-02-22 1994-09-01 Alza Corporation Compositions destinees a l'apport oral d'agents actifs
US6096863A (en) * 1996-08-23 2000-08-01 Regents Of The University Of Minnesota Self-assembling amphiphiles for construction of peptide secondary structures
EP0963196B1 (fr) 1996-09-27 2005-01-19 Jagotec Ag Systeme hyaluronique d'administration de medicaments
US20030180348A1 (en) * 2002-03-22 2003-09-25 Levinson R. Saul Transcellular drug delivery system
JP5695308B2 (ja) * 2009-10-02 2015-04-01 株式会社フェース コラーゲン修飾リポソームからなる化粧料基剤およびそれを含有する皮膚化粧料
WO2011083845A1 (fr) 2010-01-08 2011-07-14 富士フイルム株式会社 Agent de ciblage pour site tumoral
JP5756602B2 (ja) * 2010-04-16 2015-07-29 株式会社フェース ゼラチンおよび／またはエラスチン構成ポリペプチドで修飾されたリポソームからなる化粧料基剤およびそれを含有する皮膚化粧料

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5000960A (en) * 1987-03-13 1991-03-19 Micro-Pak, Inc. Protein coupling to lipid vesicles
GB8713747D0 (en) * 1987-06-12 1987-07-15 Unilever Plc Skin treatment composition
US5064655A (en) * 1989-02-24 1991-11-12 Liposome Technology, Inc. Liposome gel composition and method
AU652778B2 (en) * 1990-10-15 1994-09-08 Quest International B.V. Treatment composition
EP0525167B1 (fr) * 1991-02-14 1995-09-13 Baxter International Inc. Liberation soutenue de medicaments par application topique de liposomes bioadhesifs

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9214446A1 *

Also Published As

Publication number Publication date
AU1259292A (en) 1992-09-15
CA2079450A1 (fr) 1992-08-15
AU656173B2 (en) 1995-01-27
WO1992014446A1 (fr) 1992-09-03
JPH05506040A (ja) 1993-09-02

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