WO1999036056A1 - Preparation de lipomatrice - Google Patents

Preparation de lipomatrice Download PDF

Info

Publication number
WO1999036056A1
WO1999036056A1 PCT/US1999/000924 US9900924W WO9936056A1 WO 1999036056 A1 WO1999036056 A1 WO 1999036056A1 US 9900924 W US9900924 W US 9900924W WO 9936056 A1 WO9936056 A1 WO 9936056A1
Authority
WO
WIPO (PCT)
Prior art keywords
lipomatrix
lipid
liposomes
antigen
vaccine
Prior art date
Application number
PCT/US1999/000924
Other languages
English (en)
Inventor
Lawrence Boni
Michael M. Batenjany
Mary E. Neville
Richard J. Robb
Mircea C. Popescu
Original Assignee
Biomira Usa 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 Biomira Usa Inc. filed Critical Biomira Usa Inc.
Priority to JP2000539831A priority Critical patent/JP2002509102A/ja
Priority to AU22316/99A priority patent/AU756570B2/en
Priority to CA002318556A priority patent/CA2318556A1/fr
Priority to EP99902298A priority patent/EP1045688A1/fr
Publication of WO1999036056A1 publication Critical patent/WO1999036056A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • 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
    • A61K9/1277—Processes for preparing; Proliposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001169—Tumor associated carbohydrates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • Liposomes are increasingly important as vehicles for the delivery of pharmaceutical agents. In addition to such applications, their use as immunologic adjuvants is especially important.
  • the lipid mixture in an organic solvent is dried to a thin film using a rotovap.
  • An aqueous phase usually containing a solute to be encapsulated, is then added with vortexing to form liposomes.
  • an aqueous phase with solute to be entrapped, is added to the organic phase which is subsequently removed by either vacuum or sparging with an inert gas, thus forming liposomes.
  • the known methods have significant technical, economic and environmental drawbacks. Specifically, the thin film method can not be readily scaled up. Moreover, in the mixed organic/aqueous phase systems, the removal of the organic phase is cumbersome and often incomplete. Thus, the final product will contain residual organic solvent, which is potentially toxic and carcinogenic. Indeed, both methods typically employ such toxic substances, e.g. , chloroform, acetonitrile and acetone.
  • a Lipomatrix composition is provided, which forms liposomes only upon rehydration. This composition is particularly suited to the manufacture of novel tumor vaccines that induce superior immune responses.
  • Lipomatrix preparation which (a) can be scaled up, (b) will not contain unacceptable residual solvents and (c) is quick and simple from a manufacturing standpoint. Further to these and other objects, methods are provided for economically and safely producing a Lipomatrix, which can be used to make liposomes that are suitable for a wide variety of biomedical uses, and especially for vaccine applications.
  • a method of preparing a Lipomatrix where a water miscible organic phase, containing a phospholipid and at least one other lipid is mixed with an aqueous phase in a ratio of from about 100: 1 to about 5: 1 (v/v), then drying the mixture.
  • Another embodiment of the invention provides a Lipomatrix containing between about 20 Mol% and about 60 Mol% cholesterol.
  • a method of preparing liposomes wherein the inventive Lipomatrix is rehydrated.
  • a vaccine is provided which comprises liposomes prepared from a Lipomatrix according to the methods of the invention.
  • Figure 1 is a schematic illustrating the general process for preparing a Lipomatrix according to the invention.
  • Figure 2 shows the fluorescence emission spectra of carboxyfluorescein entrapped in a Lipomatrix compared to what would be expected for 100% entrapment.
  • Figures 3A and 3B are electron micrographs.
  • Figure 3 A shows the absence of liposomal structures in a typical Lipomatrix before lyophilization and
  • Figure 3B shows the formation of liposomes after hydration of the Lipomatrix.
  • Figures 4A and 4B show interferon gamma (IFN- ⁇ ) production of mouse lymph node cells ( Figure 4A) and spleen cells (Figure 4B) in response to antigen challenge.
  • the mice were immunized with liposomal MUC-1 prepared by hydrating the Lipomatrix formulations described in Example 3.
  • Figure 5 shows interferon gamma (IFN- ⁇ ) production of mouse lymph node cells and spleen cells in response to antigen challenge.
  • the mice were immunized with a liposomal MUC-1 vaccine prepared by hydrating Lipomatrix formulations containing varying amounts of cholesterol as described in Example 4.
  • Figure 6 compares the differential scanning calorimetry (DSC) heating scans of liposomal MUC-1 preparations made by hydrating a Lipomatrix formulated at 50 Mol% cholesterol and that of DPPC liposomes at the same bulk lipid concentration (20 mg/mL).
  • DSC differential scanning calorimetry
  • Figure 7 shows the Raman vibrational spectroscopic profile of a Lipomatrix formulation prepared as described in Example 1.
  • solid and dotted lines represent two different sites in the lyophilized film.
  • Panel B shows a Raman profile for the hydrated formulation.
  • Figure 8 shows the interferon gamma (IFN- ⁇ ) production by lymph node cells and spleen cells in response to antigen challenge after immunization with a liposomal MUC-1 vaccine prepared from a Lipomatrix. Time points represent the hours that the hydrated Lipomatrix formulation stood at room temperature before subcutaneous injection into mice. DETAILED DESCRIPTION OF THE INVENTION
  • the invention provides a Lipomatrix, which is essentially comprised of lipid lattices (or stacked bilayers), which forms liposomes upon hydration.
  • the invention also provides a simplified method for producing highly effective liposome preparations, by hydration of a Lipomatrix. This method of producing liposomes from a Lipomatrix overcomes many of the above-described obstacles to the efficient and safe manufacture of liposomes. In a specific example, the usefulness of the Lipomatrix is demonstrated in the manufacture of a mucin-based cancer vaccine.
  • the inventive Lipomatrix is essentially a dried (e.g. , lyophilized) composition of lipid that is capable of forming liposomes upon reconstitution.
  • the composition is characterized as a matrix of stacked bilayers, and is essentially liposome-free. It is comprised in its dried form of mostly lipid, with at most trace amounts of solvent and less than about 5 percent water by weight. Trace amounts of solvent are generally less than about 0.1 % and typically less than about 0.05% by weight.
  • Some exemplary compositions contain (by weight) about 93-94% lipid and some contain less than about 3-4% water.
  • the Lipomatrix Prior to drying, the Lipomatrix exists as an essentially liposome-free suspension of lipid. In this state, the Lipomatrix is comprised mostly of solvent and water, with lipid levels usually being less than about 10 percent, by weight. In many cases, however, the lipid will be present at even lower levels, such as less than about five percent by weight. Some exemplary compositions have lipid at levels less than about 1.5% by weight. An embodiment below utilizes lipid levels of about 0.9%. Solvent levels will usually be kept above about 80% and in some instances may be about 95 % . Some exemplary compositions have between about 85% and about 90%, while others will be above about 90%. On the other hand, water is usually present at somewhat lower levels, typically less than about 20%. Many compositions have between about 10% and about 15% water by weight.
  • the Lipomatrix compositions In suspension (prior to drying), some of the Lipomatrix compositions have solvent:water ratios of from about 5: 1 to about 20: 1 (vol ./vol.); two exemplary compositions have ratios of 7:1 and 9:1.
  • the lipid:solvent mass ratio should be above about 1:20, and it usually will be less than about 1:50. The higher end of this range (e.g. , from about 1: 35 to 1: 50) is preferred for its ability to produce superior vaccines.
  • the basic method involves first creating an organic phase, by dissolving appropriate lipids in a water miscible organic solvent, which is mixed with a small volume of an aqueous phase to induce molecular ordering, i.e.
  • Lipomatrix formation of the Lipomatrix.
  • An optional sterilizing filtration step is included either before or after this mixing.
  • the resulting Lipomatrix is lyophilized or freeze-dried.
  • the dried mixture can be hydrated in an appropriate medium, thereby spontaneously forming liposomes capable of entrapping an aqueous solute.
  • liposomes are not appreciably formed in the present method at any time prior to hydration. Indeed, as demonstrated below in the Examples, no liposomal structures are detected prior to hydration.
  • the instant methods do not involve pre-forming liposomes that are dried and merely rehydrated by the addition of water.
  • the Lipomatrix preparation is essentially liposome-free until it is hydrated, as set out below.
  • the instant methods can be used to prepare a Lipomatrix, which can be hydrated to make highly effective liposome compositions that have a high cholesterol content. It is believed that the added cholesterol broadens the transition temperature and eliminates domains of either phospholipid, cholesterol or other lipophilic components and their combinations. See Ladbroke et al., Biochim. Biophys. Acta 150:333-40 (1968). These previous studies employed cholesterol in liposomes to decrease leakage of solute from the liposome, for modifying liposome size or for mimicking plasma membrane compositions. The art did not recognize, however, that certain liposomes containing high cholesterol content have superior adjuvant properties. As demonstrated below in the Examples, when such liposomes are prepared from the inventive Lipomatrix, they have surprisingly better immuno- stimulatory properties.
  • the present methods for preparing Lipomatrix involve, first, preparing an organic phase by dissolving at least one lipid in a water-miscible organic solvent.
  • Suitable lipids include phospholipids, in particular lecithins, phosphatidylglycerols, phosphatidylethanolamines, phosphatidylserines and other natural and synthetic compounds known in the art. See, for example, WO 91/04019 (1991) at pages 8 and 9.
  • Preferred phospholipids specifically include dimyristoyl phosphatidylcholine (DMPC), dipalmitoyl phosphatidylcholine (DPPC) and dimyristoyl phosphatidylglycerol (DMPG).
  • Other suitable lipids include sterols, and especially cholesterol.
  • glycolipids and lipid adjuvants such as monophosphoryl Lipid A (MPL) or Lipid A.
  • the final concentration of phospholipids is between 15 mg/mL and about 36 mg/mL, depending of course on the solubility of the particular lipid(s) used in the solvent chosen.
  • Mild heating for example between 50 °C to about 60 °C, may also be employed in the Lipomatrix formation. The degree of heating optionally used will depend in large part on the solubility and stability of the various organic phase components.
  • the inventive Lipomatrix is particularly useful when manufactured with high cholesterol content.
  • the inventive methods can include in the phospholipid matrix about 20 Mol% or more of cholesterol.
  • Some preferred Lipomatrix formulations contain from about 30 Mol% to about 60 Mol% cholesterol.
  • the organic solvent should be water miscible and is usually an aliphatic alcohol.
  • Preferred aliphatic alcohols include, but are not limited to, ethanol and tert-butanol.
  • Other organic solvents, and especially other alcohols, may be employed. They should, however, be amenable to drying by, for example, lyophilization, and they should be non-toxic. Thus, solvents such as those typically employed in the thin film method are generally unacceptable.
  • an aqueous phase is provided.
  • the aqueous phase may contain one or more buffers, salts, and bulking agents.
  • Preferred bulking agents include sugars, and preferred sugars include mannitol.
  • Appropriate buffers, salts and bulking agents preferably are physiologically compatible and are widely known to those in the art. Of course, the concentration of these buffers, salts and bulking agents chosen will depend primarily on physiological compatibility, will be understood by those in the art.
  • the organic phase and the aqueous phase are either mixed and optionally sterile filtered, or optionally sterile filtered separately and then mixed. When mixing, the ratio of organic phase to aqueous phase is preferably from about 7: 1 to about 9:1 volume/ volume.
  • This ratio could be as high as about 100: 1 and as low as about 5:1 organic: aqueous. Ranges such as from about 20:1 to about 6: 1 are also acceptable.
  • the lipid: solvent mass ratio should be between 1:20 and about 1:50, with the higher end of that range being preferred (e.g. , from about 1:35 to 1:50).
  • the mixing can be done at ambient temperatures, but may be done at temperatures as high as the highest melting temperature of the lipids employed. At this stage, although molecular ordering occurs and open bilayers are formed (i.e. , Lipomatrix), liposomes are not detectable.
  • the Lipomatrix can be dried by lyophilization or other suitable means.
  • the Lipomatrix may be dried in bulk. Prior to drying, it can be divided into aliquots of a suitable size. Typically, the Lipomatrix solution is aliquoted into vials with continuous mixing, followed by lyophilization. The resulting Lipomatrix formulation is stable and suitable for storage.
  • liposomes are accomplished when the dried Lipomatrix film or cake is hydrated with a suitable aqueous solvent, such as water, saline, or an appropriate buffer, optionally containing a solute for encapsulation.
  • a suitable aqueous solvent such as water, saline, or an appropriate buffer, optionally containing a solute for encapsulation.
  • the temperature of the hydration solution may be ambient to above the transition temperature of the highest melting lipid. Only upon hydration, are liposomes formed which are capable of entrapping an aqueous solute. This is a significant simplification over the art, which relies on pre-forming liposomes prior to drying. Moreover, because the prior art methods relied on such pre-formation, stabilizers were needed to maintain the integrity of the liposomes. Such stabilizers are unnecessary in the instant methods because liposomes are not formed prior to hydration.
  • Lipomatrix prepared according to the invention can be used in a wide variety of applications after hydrating to form liposomes, especially biomedical applications. For example, they can be used to deliver a wide variety of pharmacologically active agents.
  • lipophilic agents for example hydrophobic peptides
  • hydrophilic pharmacologically active agents may be entrapped within the resultant liposomes upon hydration. Charged molecules might be electrostatically associated with the phospholipids of the liposomes. Examples of suitable lipophilic and hydrophilic pharmacological agents can be found inffy et al. , U.S. Patent No.
  • pharmacologically active agents include, for example, adjuvants, cytokines, antibodies and any other known pharmaceuticals.
  • cytokines such as interleukin-2 (IL-2), granulocyte- macrophage colony stimulating factor (GM-CSF) and interferon-gamma (IFN- ⁇ ), which may be used alone or in conjunction with other agents. Combinations of any of these agents are also envisioned.
  • IL-2 interleukin-2
  • GM-CSF granulocyte- macrophage colony stimulating factor
  • IFN- ⁇ interferon-gamma
  • the inventive methods are especially useful in the manufacture of vaccines.
  • Tumor antigens may be derived, for example, from lung cancer, colon cancer, melanoma, neuroblastoma, breast cancer, ovarian cancer and the like.
  • a preferred tumor antigen is MUC-1 and related antigenic peptides.
  • MUC-1 mucin is a high molecular weight glycoprotein with a protein core consisting of tandem repeats of a 20 amino acid sequence and highly-branched carbohydrate side chains.
  • Many human adenocarcinomas, such as breast, colon, lung, ovarian and pancreatic cancers abundantly over-express and secrete underglycosylated MUC-1 protein.
  • MUC-1 mucin expression
  • MUC-1 is, therefore, a clinically significant marker for these cancers.
  • Particularly useful antigenic MUC-1 peptide derivatives are based on the 20 amino acid repeat sequence.
  • other clinically relevant antigens include allergens, viral antigens, bacterial antigens and antigens derived from parasites.
  • Antigens are usually macromolecules such as peptides, lipids, carbohydrates and combinations thereof which may simply be mixed together or covalently linked, as in glycopeptides, glycolipids.
  • Typical vaccine compositions comprise liposomes hydrated from the inventive Lipomatrix formulations containing an antigen, such as a tumor antigen. Additionally, vaccine compositions may contain one or more immunomodulators.
  • An immunomodulator is any substance that alters the immune response, and preferably stimulates the antigenic immune response.
  • Typical immunomodulators include adjuvants, such as monophosphoryl Lipid A and Lipid A.
  • Other immunomodulators include lymphokines and cytokines, and specifically interleukins, in particular IL-2.
  • the vaccines are typically formulated using a pharmaceutically acceptable excipient.
  • excipients are well known in the art, but typically will be a physiologically tolerable aqueous solution. Physiologically tolerable solutions are those which are essentially non-toxic. Preferred excipients will either be inert or enhancing with respect to antigenic activity.
  • an anti-tumor vaccine is prepared which comprises liposomes containing a synthetic MUC-1 peptide, a tumor antigen, and using monophosphoryl Lipid A (MPL) or Lipid A as an immunomodulatory adjuvant. See Koganty et al. , DDT 1 : 190-98 (1996); Alving et al.
  • the MUC-1 peptide is a synthetic peptide version with antigenic properties similar to the parent MUC-1 glycoprotein.
  • a vaccine formulated with this peptide antigen is currently under clinical investigation. See Koganty et al. DDT 1: 190-198.
  • Various formulations were tested in mice for the induction of an antigen-specific T cell response.
  • BP1-148 lipopeptide has the following structure: NH 2 -[STAPPAHGVTSAPDTRPAPGSTAPP(K-lipid conjugated)G]-COOH.
  • the following amounts of the warmed stock solutions were added to a clean stoppered 5 mL glass vial: 49.1 ⁇ L of DPPC, 103.5 ⁇ L cholesterol, 60 ⁇ L of BP1- 148, 30 ⁇ L Lipid A and 657.4 ⁇ L of absolute ethanol.
  • the mixture was vortexed briefly (3 seconds x 7 times) and returned to the 55 °C water bath.
  • One hundred microliters of deionized water (55 °C) was added into the vial and the was mixture vortexed briefly as above.
  • the mixture was returned to the 55 °C water bath for 15-20 minutes, vortexing (as above) twice during that period.
  • the vials were cooled to room temperature, placed in a Dura-Stop MP shelf lyophilizer (FTS Systems, Stone Ridge, NY).
  • This example shows the effectiveness of various liposome preparations made from a Lipomatrix by the instant method in generating an immune response.
  • the Lipomatrix was prepared, as outlined, by adding nine parts of an ethanol solution containing lipid and lipopeptide to one part water.
  • the resultant liposomes contained, per 0.1 mL dose, 10 ⁇ g
  • MUC-1 peptide See, e.g. , Koganty et al. , DDT 1: 190-98 (1996)), 20 ⁇ g MPL and:
  • MB-IX-2 2 mg DPPC; MB-IX-3. 1.86 mg DMPC, 0.14 mg DMPG;
  • MB-IX-5 1.63 mg DPPC, 0.37 mg cholesterol; or
  • MB-IX-6 1.6 mg DPPC, 0.125 mg DMPG, 0.375 mg cholesterol.
  • mice were immunized by subcutaneous injection in the inguinal area and sacrificed nine days later. Lymph nodes and spleens were removed. Lymph node T cells were purified by passing through a nylon wool column. For the lymph node assay, Antigen
  • APCs Presenting Cells
  • lymph node cells with APCs
  • spleen cells were incubated for four days with appropriate peptide antigens, followed by a 24 hour incubation with Alamar Blue, after which the OD ratio at 610 to 570 nm was measured.
  • Alamar Blue Prior to the addition of Alamar Blue, supernatants of the cell proliferation assay were harvested and the gamma-interferon was measured in an Immuno-Fluorescence Assay (IF A). See Ahmen et al. , J. Immunol. Methods 170:211-224.
  • DPPC/cholesterol formulations induce a strong immune response with respect to lymphocyte IFN-gamma production.
  • the response was dependent on the Mol % cholesterol in the formulation, with no biological response for preparations containing 10 or 20 Mol% cholesterol.
  • This example shows the uniformity of hydrated preparations made using the Lipomatrix process prepared with an ethanol: water volume ratio of 9: 1. Preparations were analyzed using differential scanning calorimetry (DSC) and Raman vibrational spectroscopy. The final product contained 13.1 mg/mL DPPC, 6.9 mg/mL cholesterol (i.e. , 50 Mol%), 200 ⁇ g/mL Lipid A and 400 ⁇ g/mL BP1-148.
  • DSC runs were performed on a Hart (now CSC) Scientific Model 7707 series differential scanning microcalorimeter (Provo, UT) at 60 °C/hr. Fresh aliquots were used for each time point from a single sample vial hydrated at 55 °C. Each run included a cell containing normal saline solution for baseline determination. After baseline subtraction and correction for the thermal instrument response, calorimetric data were analyzed to yield excess heat content ( ⁇ Watts) as a function of temperature, using software supplied by Hart Scientific. The calorimetric data were imported into Grams/32, v.5.0 (Galactic Industries Corporation, Salem, NH) for baseline and offset correction, smoothing and plotting.
  • Raman spectra were again collected using a x50 objective, but the laser was defocused 80 % to prevent local heating of the bilayer structures. In both cases, power at the laser head was set to 300 mW and reduced to 25 % at the microscope.
  • the Raman signal was dispersed by the spectrometer (1800gr/mm grating) onto a CCD detector. Typically 10 spectra were coadded using a time constant of 30 - 60 seconds per collection. Spectral resolution was at ⁇ 1 cm "1 .
  • the DSC profile for a liposomal preparation made by the Lipomatrix process as outlined in Example 1 revealed a very flat endotherm that did not change in time.
  • the DSC heating profile of liposomes comprised of DPPC alone exhibits two prominent transitions at 38 °C (pretransition) and 41 °C (main transition).
  • the absence of these transitions in the Lipomatrix formulations at 50 Mol% cholesterol indicates that the components are devoid of DPPC-rich domains and is indicative of a liposomal preparation in which the components are well mixed.
  • Raman vibrational spectroscopy was used.
  • Figure 7 A shows that at two different sites (solid and dotted lines) in the lyophilized film there was no significant difference in the relative concentrations of cholesterol to DPPC. Moreover, the same relative ratios of cholesterol to DPPC was also observed in the hydrated product (Figure 7B).
  • a 120 mL batch was prepared by the Lipomatrix process of Liposomal MUC-1 vaccine at 15 mg bulk lipid (at 50 Mol% cholesterol), 300 ⁇ g BP1-148, 150 ⁇ g Lipid A per vial as outlined in example 1 and filtered at room temperature one hour after production (MB- XLIV-B) and eight hours after production (MB-XLIV-A). As shown in the tables below, no detectable losses were observed by HPLC. HPLC Results of 120 mL Scale-up Formulation (MB-XLIV-A and MB-XLIV-B)
  • Example 2 illustrates the use of the present Lipomatrix formulations in preparing a tumor antigen-specific cancer vaccine.
  • a Liposomal MUC-1 vaccine was prepared, as in Example 1, which contained 15 mg bulk lipid (at 50 Mol% cholesterol), 300 ⁇ g BP1-148, 150 ⁇ g Lipid A per vial. The following parameters were varied: a. the alcohol, ethanol or tert-butanol; b. the solvent to water ratio; and c. the lipid to solvent mass ratio.
  • This example exhibits the stability of the lyophilized Lipomatrix.
  • a liposomal MUC-1 vaccine was prepared, which had 15 mg bulk lipid (at 50 Mol% cholesterol), 300 ⁇ g BP1-148, 150 ⁇ g Lipid A per vial by the method of Example 1, and analyzed at time zero and after 3 and 6 months to determine the stability by HPLC analysis.
  • the Designations A and B are as defined in Example 6. As shown below, no significant changes were observed from the initial time point:
  • Lipomatrix formulations are stable at room temperature.
  • the Lipomatrix process was used to make several vials of Liposomal MUC-1 vaccine with 15 mg bulk lipid (at 50 Mol% cholesterol), 150 ⁇ g BP1-148, 150 ⁇ g Lipid A per vial, using an ethanol solventiwater ratio of 9: 1 by volume.
  • mice were subcutaneously injected with these preparations after room temperature storage of the hydrated preparations for 0, 2, 4 and 24 hours. Two sets of four mice were injected at each time point. The averages shown in Figure 8 demonstrate a stable product following hydration at all time points.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Immunology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Oncology (AREA)
  • Molecular Biology (AREA)
  • Dispersion Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicinal Preparation (AREA)

Abstract

La présente invention concerne une lipomatrice composée de réseaux lipidiques à doubles couches empilées qui forment des liposomes lorsqu'elles sont hydratées, et un procédé simplifié permettant de fabriquer des préparations de liposomes à haute efficacité. L'invention se rapporte en outre à des compositions de vaccin dotées de propriétés immunologiques supérieures, dans lesquelles sont utilisés des liposomes biomédicaux obtenus à partir d'une lipomatrice selon des procédés sûrs et efficaces. L'utilisation des procédés de l'invention permet de produire des vaccins extrêmement actifs contre les antigènes tumoraux.
PCT/US1999/000924 1998-01-16 1999-01-15 Preparation de lipomatrice WO1999036056A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
JP2000539831A JP2002509102A (ja) 1998-01-16 1999-01-15 リポマトリックス製剤
AU22316/99A AU756570B2 (en) 1998-01-16 1999-01-15 Lipomatrix preparation
CA002318556A CA2318556A1 (fr) 1998-01-16 1999-01-15 Preparation de lipomatrice
EP99902298A EP1045688A1 (fr) 1998-01-16 1999-01-15 Preparation de lipomatrice

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US7168898P 1998-01-16 1998-01-16
US60/071,688 1998-01-16

Publications (1)

Publication Number Publication Date
WO1999036056A1 true WO1999036056A1 (fr) 1999-07-22

Family

ID=22102934

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1999/000924 WO1999036056A1 (fr) 1998-01-16 1999-01-15 Preparation de lipomatrice

Country Status (5)

Country Link
EP (1) EP1045688A1 (fr)
JP (1) JP2002509102A (fr)
AU (1) AU756570B2 (fr)
CA (1) CA2318556A1 (fr)
WO (1) WO1999036056A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002043699A2 (fr) * 2000-12-01 2002-06-06 Biomira, Inc. Preparation de liposomes de grande taille par perfusion dans de l'ethylene glycole
WO2006076891A2 (fr) * 2005-01-18 2006-07-27 Abnoba Heilmittel Gmbh Procede et dispositif d'encapsulage de substances dans des liposomes ayant une structure membrane librement ajustable
US8951450B2 (en) 2009-09-02 2015-02-10 Biomedcore, Inc. Apparatus and method for production of liposomes
US9173929B2 (en) 2004-04-01 2015-11-03 Oncothyreon Inc. Mucinous glycoprotein (MUC-1) vaccine

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006002703A1 (fr) * 2004-07-01 2006-01-12 Basf Aktiengesellschaft Procede de production d'acide acrylique par oxydation partielle en phase gazeuse a catalyse heterogene de propylene
EP2367532A4 (fr) * 2008-12-17 2012-12-12 Oncothyreon Inc Procédé de fabrication de petits liposomes

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2157172A (en) * 1984-04-09 1985-10-23 Sandoz Ltd Interleukin therapy
WO1992011842A1 (fr) * 1991-01-14 1992-07-23 The Board Of Regents, The University Of Texas System Poudre preliposomique de polyene aux liposomes et son procede de preparation
WO1995004523A1 (fr) * 1993-08-06 1995-02-16 Opperbas Holding B.V. Methode de preparation de vesicules chargees de structures biologiques, de biopolymeres et/ou d'oligomeres
WO1996039121A1 (fr) * 1995-06-06 1996-12-12 Board Of Regents, The University Of Texas System Suspensions liposomales submicroniques obtenues a partir de lyophilisats preliposomaux
WO1997029769A1 (fr) * 1996-02-16 1997-08-21 Biomira Usa Inc. Vaccin contre des malignites affectant les lymphocytes b

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2157172A (en) * 1984-04-09 1985-10-23 Sandoz Ltd Interleukin therapy
WO1992011842A1 (fr) * 1991-01-14 1992-07-23 The Board Of Regents, The University Of Texas System Poudre preliposomique de polyene aux liposomes et son procede de preparation
WO1995004523A1 (fr) * 1993-08-06 1995-02-16 Opperbas Holding B.V. Methode de preparation de vesicules chargees de structures biologiques, de biopolymeres et/ou d'oligomeres
WO1996039121A1 (fr) * 1995-06-06 1996-12-12 Board Of Regents, The University Of Texas System Suspensions liposomales submicroniques obtenues a partir de lyophilisats preliposomaux
WO1997029769A1 (fr) * 1996-02-16 1997-08-21 Biomira Usa Inc. Vaccin contre des malignites affectant les lymphocytes b

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002043699A2 (fr) * 2000-12-01 2002-06-06 Biomira, Inc. Preparation de liposomes de grande taille par perfusion dans de l'ethylene glycole
WO2002043699A3 (fr) * 2000-12-01 2002-10-17 Lawrence Boni Preparation de liposomes de grande taille par perfusion dans de l'ethylene glycole
US9173929B2 (en) 2004-04-01 2015-11-03 Oncothyreon Inc. Mucinous glycoprotein (MUC-1) vaccine
WO2006076891A2 (fr) * 2005-01-18 2006-07-27 Abnoba Heilmittel Gmbh Procede et dispositif d'encapsulage de substances dans des liposomes ayant une structure membrane librement ajustable
WO2006076891A3 (fr) * 2005-01-18 2006-11-09 Abnoba Heilmittel Gmbh Procede et dispositif d'encapsulage de substances dans des liposomes ayant une structure membrane librement ajustable
US8951450B2 (en) 2009-09-02 2015-02-10 Biomedcore, Inc. Apparatus and method for production of liposomes

Also Published As

Publication number Publication date
EP1045688A1 (fr) 2000-10-25
AU756570B2 (en) 2003-01-16
JP2002509102A (ja) 2002-03-26
AU2231699A (en) 1999-08-02
CA2318556A1 (fr) 1999-07-22

Similar Documents

Publication Publication Date Title
US20020051813A1 (en) Lipomatrix preparation
AU631377B2 (en) Affinity associated vaccine
RU2216315C2 (ru) Способ получения липосом
FI86371C (fi) Foerfarande foer framstaellning av farmaceutiska liposomkompositioner.
US5466468A (en) Parenterally administrable liposome formulation comprising synthetic lipids
JPS6339810A (ja) リポソ−ムの製造法
JPH03169808A (ja) 新しい脂質微粒子の製法
JPH09501169A (ja) 小胞に生体高分子物質を高度に充填する方法
EA011881B1 (ru) Лиофилизация виросом
US9603799B2 (en) Liposomal vaccine adjuvants and methods of making and using same
EP0831780B1 (fr) Formulation liposomale amelioree
CN1044093C (zh) 低毒性药物类脂制剂
US5270053A (en) Parenterally administerable liposome formulation comprising synthetic lipid
EP0356340B1 (fr) Vaccin à affinité associée
AU756570B2 (en) Lipomatrix preparation
Kersten et al. Immunogenicity of liposomes and iscoms containing the major outer membrane protein of Neisseria gonorrhoeae: influence of protein content and liposomal bilayer composition
AU740619B2 (en) Aqueous pharmaceutical composition comprising an active ingredient which is highly insoluble in water
Chattaraj et al. Physicochemical characterization of influenza viral vaccine loaded surfactant vesicles
CA2039477C (fr) Formule a base de liposomes devant etre administree par voie parenterale, et comprenant des lipides synthetiques
EP0713387B1 (fr) Methode de preparation de vesicules chargees de structures biologiques, de biopolymeres et/ou d'oligomeres
CA2168656C (fr) Methode de preparation de vesicules chargees de structures biologiques, de biopolymeres et (ou) d'oligomeres
US6207170B1 (en) Patient-specific white blood cell malignancy vaccine from membrane-proteoliposomes
WO1990001947A1 (fr) Vaccin a affinite associee
Haensler Liposomal adjuvants: preparation and formulation with antigens
Gregoriadis et al. The immunoadjuvant action of liposomes: role of structural characteristics

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GD GE HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 1999902298

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2318556

Country of ref document: CA

Ref country code: CA

Ref document number: 2318556

Kind code of ref document: A

Format of ref document f/p: F

NENP Non-entry into the national phase

Ref country code: KR

ENP Entry into the national phase

Ref country code: JP

Ref document number: 2000 539831

Kind code of ref document: A

Format of ref document f/p: F

WWE Wipo information: entry into national phase

Ref document number: 22316/99

Country of ref document: AU

WWP Wipo information: published in national office

Ref document number: 1999902298

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWG Wipo information: grant in national office

Ref document number: 22316/99

Country of ref document: AU

WWW Wipo information: withdrawn in national office

Ref document number: 1999902298

Country of ref document: EP