WO1993004672A1 - Procede de renforcement du systeme immunitaire d'un hote a l'aide de polypeptides encapsules dans des liposomes - Google Patents

Procede de renforcement du systeme immunitaire d'un hote a l'aide de polypeptides encapsules dans des liposomes Download PDF

Info

Publication number
WO1993004672A1
WO1993004672A1 PCT/US1992/007562 US9207562W WO9304672A1 WO 1993004672 A1 WO1993004672 A1 WO 1993004672A1 US 9207562 W US9207562 W US 9207562W WO 9304672 A1 WO9304672 A1 WO 9304672A1
Authority
WO
WIPO (PCT)
Prior art keywords
growth factor
liposome
somatotropin
animal
encapsulated
Prior art date
Application number
PCT/US1992/007562
Other languages
English (en)
Inventor
Carl K. Edwards, Iii
Aleksander Blum
Original Assignee
Pitman-Moore, 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 Pitman-Moore, Inc. filed Critical Pitman-Moore, Inc.
Publication of WO1993004672A1 publication Critical patent/WO1993004672A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/27—Growth hormone [GH] (Somatotropin)
    • 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
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • 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
    • 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
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55555—Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers

Definitions

  • This invention relates to a method for enhancing the immune system of an animal by encapsulating a polypeptide into liposomes and administering the encapsulated polypeptide to an animal in sufficient quantity to enhance the animal's immune system against a pathogen. More specifically, the invention relates to a method of encapsulating a polypeptide into multilamellar, unilamellar or any other suitable liposome and parenterally administering the liposome- encapsulated polypeptide to an animal in sufficient quantity to enhance the animal's immune system against pathogenic gram negative bacteria.
  • Some gram-negative bacteria such as the Salmonella species, are facultative intracellular bacteria that are usually pathogenic to man and other mammals. In humans, the most common diseases caused by the Salmonella species are typhoid fever and gastroenteritis. Salmonella bacteria are capable of surviving within the phagocytic cells of the immune system. However, phagocytic cells such as macrophages (M ) long have been known to be particularly essential for host defense against Salmonella species. M ⁇ must be exposed to T cell-derived lymphokines such as interferon-c (IFN-c) and granulocyte-macrophage colony stimulating factor (GM-CSF) to fully exert their antibacterial activity.
  • IFN-c interferon-c
  • GM-CSF granulocyte-macrophage colony stimulating factor
  • growth hormone i.e., somatotropin
  • prolactin also have been discovered to enhance a host's immune system against various pathogens by increasing the number of macrophages and by priming the macrophages to release reactive oxygen intermediates (see U.S. Patent No. 4,837,202).
  • the immune enhancing effect of somatotropin has been tested in laboratory rats infected with the bacterium Salmonella typhimurlum.
  • the ability of hypophysectomized rats, i.e., rats that have had their pituitary gland (source of growth hormone) experimentally excised, to survive Salmonella typhimurlum infections has been compared to that of rats having their pituitary intact.
  • hypophysectomized rats all die within a few days, while the pituitary-intact rats survive for a significantly (as assessed by chi square analyses) longer period of time.
  • the dose of Salmonella typhimurlum is reduced and the hypophysectomized rats are treated with somatotropin, IFN-c or tetracycline, the survival rate is enhanced.
  • peritoneal M ⁇ from hypophysectomized rats that were infected in vitro with Salmonella typhimurium only killed half the number of extracellular bacteria as compared to pituitary-intact rats.
  • somatotropin In addition to Salmonella species, somatotropin also has been shown to induce alveolar M ⁇ against respiratory pathogens such as Pasteurella multoclda , a gram negative bacterium. Porcine alveolar M ⁇ treated in vitro for 18 hours with either native porcine somatotropin or recombinant porcine IFN-c killed opsonized extracellular Pasteurella multoclda after a 4 hour period. Addition of 10 8 viable Pasteurella multocida to untreated porcine alveolar M ⁇ resulted in 14 ⁇ 5xl0 6 colony forming units (CFU).
  • CFU colony forming units
  • porcine alveolar M ⁇ Treatment of the porcine alveolar M ⁇ with native porcine somatotropin or recombinant porcine IFN-c reduced the CFU by 95% and 94% respectively (C.K. Edwards, III, et al., "Growth Hormone Enhances Bacterial Killing By Alveolar Macrophages", 1990 ASBMB/AA1, NIH AG06246, USDA 89- 37265-4536). These results imply a potential clinical application of somatotropin in enhancing a host's immune system against gram-negative bacterial infections.
  • Somatotropin is a polypeptide which upon oral ingestion is readily digested by the acids and enzymes of an animal's stomach. Also, large amounts of somatotropin can be degraded upon parenteral administration as well before the somatotropin can act on the immune system. In order for the somatotropin to be clinically effective, it desirably is incorporated into an appropriate vehicle.
  • liposomes i.e., multilamellar or unilamellar concentric lipid bilayer vesicles with layer(s) of aqueous media separating the lipid bilayer(s)
  • liposomes have been used as carriers for therapeutic agents, anticancer drugs, antifungal agents and immunomodulators.
  • Gabriel Lopez- Berestein and Royce J. Fidler "Liposomes in the Therapy of Infectious Disease and Cancer, " UCLA Symposia on Molecular and Cellular Biology, New Series, Volume 89, (1989). It also has been implied that liposomes can be useful in improving the immune response to vaccines. Biotechnology News, Vol. 10, No. 13, p.3 (1989).
  • Antimonials, amphotericin B, and pentamide encapsulated in liposomes were found to be more effective than the free drugs for the treatment of leishmaniasis in hamsters (Amitabha Mukhopadhyay, et al. "Receptor-Mediated Drug Delivery to Macrophages in Chemotherapy of Leishmaniasis,” Science, Vol. 244, 1989).
  • Liposome encapsulated muramyl dipeptide and M ⁇ - activating factor have been shown to greatly enhance the immune system in tumor-bearing mice by activating M ⁇ against the tumorous cells (Charles Pidgeon, et al., "Macrophage Activation: Synergism Between Hybridoma MAF And Poly (1) I Poly (C) Delivered By Liposomes," The Journal of Immunology, Vol. 131, No. 1, 1983; foremost J. Fidler and Alan J. Schroit, "Synergism Between Lymphokines And Muramyl Dipeptide Encapsulated In Liposomes: In situ Activation of Macrophages And Therapy of Spontaneous Cancer Metastases, " The Journal of Immunology. Vol.
  • U.S. Patent No. 4,394,448 discloses liposomes composed of phospholipid bilayers into which DNA material is encapsulated. The liposome-encapsulated DNA then is used to insert the DNA into a host cell by contacting the membrane of the target cell with the liposome. The liposome is taken up by the cell through fusion of the liposome with the cellular membrane or by endocytosis. The liposome protects the DNA from degradation during the insertion process.
  • U.S. Patent Nos. 4,721,612 and 4,891,208 disclose liposomes composed of bilayers comprising the salt form of an organic acid derivative of a sterol such as the tris-salt form of a sterol hemisuccinate.
  • the sterol hemisuccinate liposome can entrap bioactive agents of limited solubility, such as growth hormone.
  • Bovine growth hormone encapsulated in the sterol hemisuccinate liposome can be administered intramuscularly to cows to initiate growth or increase milk production.
  • U.S. Patent No. 4,708,861 discloses liposomes containing a bioactive agent wherein the liposomes are dispersed within a gel-matrix.
  • the gel-matrix provides for prolonged release of the liposome-entrapped bioactive agent.
  • the liposome bilayer can be composed of phospholipids and related chemical structures as well as steroids, such as cholesterol.
  • the gel-matrix is composed of carbohydrates such as cellulosics. Bioactive agents such as somatotropin and other peptides and the like can be encapsulated in the liposome gel-matrix.
  • the disclosure does not teach or suggest that the liposome incorporated polypeptides can enhance an animal's immune system.
  • a polypeptide is encapsulated into liposomes, and the liposome-encapsulated polypeptide is administered to a host in sufficient quantities to enhance the host's immune system against a pathogen.
  • the liposome- encapsulated polypeptide enhances the host's immune system to a greater degree than if the polypeptide is administered to the host in non-liposome-encapsulated form.
  • the liposome-encapsulated polypeptide is especially effective in enhancing the immune system against gram-negative bacteria, particularly against Salmonella and Pasteurella species.
  • Figure 1 illustrates the survival rate of hypophysectomized (Hypox) rats infected with S.
  • FIG. 2 illustrates the survival rate of hypophysectomized (Hypox) rats (% survival vs. days after injection with S. typhimurium) infected with S. typhimurium from an experiment indicating that rats pretreated with increasing amounts of recombinant porcine somatotropin have enhanced host-protection against S. typhimurium .in vivo.
  • Figure 3 illustrates that hypophysectomized rats treated with liposomes encapsulated with recombinant porcine somatotropin have enhanced host-protection against S. typhimurium when compared to somatotropin- free liposomes or recombinant porcine somatotropin given in saline.
  • the present invention relates to a method of enhancing a host's immune system against a pathogen by administering a sufficient amount of liposome- encapsulated polypeptide to enhance phagocytic cell activity against the pathogen.
  • the liposome-encapsulated polypeptide is internalized by the phagocytic cells of the immune system, such as the polymorphonuclear leukocytes and/or the M ⁇ , when the liposome contacts the phagocytic cell's membrane and the lipid components of the liposome and the cell membrane fuse or by the process of endocytosis.
  • the polypeptide then assists the phagocytic cell to produce reactive oxygen intermediates which are toxic to many types of intracellular microbial pathogens such as Salmonella and Pasteurella species.
  • the liposomes employed to practice this invention are completely closed bilayer membranes containing an aqueous phase.
  • the liposomes may be any variety of mult la eliar vesicles (elliptical-like structures characterized by concentric membrane bilayers each separated by an aqueous layer) or unilamellar vesicles (possessing a single membrane bilayer) .
  • the size of the liposomes can vary widely. Typically, the size of the liposomes prepared according to the method of the present invention will be less than 1 micron in size. Any chemical compound capable of forming a completely closed bilayer, i.e., liposome, in aqueous solutions can be used to practice this invention.
  • amphipathic lipids can be used as constituents of the liposome bilayer.
  • Suitable hydrophilic groups include, but are not limited to, phosphatidic , sulfatidic, carboxylic and amino groups.
  • Suitable hydrophobic groups include, but are not limited to, saturated and unsaturated aliphatic hydrocarbon groups and aliphatic hydrocarbon groups substituted by at least one aromatic and/or cycloaliphatic group.
  • the preferred amphipathic compounds are phospholipids and closely related chemical structures.
  • Examples of these include, but are not limited to, phosphatidylcholine, phosphatidylethanolamine, lysolecithin, lyso- phosphatidylethanolamine, sphingomyelin, cardiolipin, phosphatidic acid, the cerebrosides, natural lecithins (e.g., egg lecithin or soybean lecithin) and synthetic lecithins such as saturated synthetic lecithins (e.g., dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine or distearoyl- phosphatidylcholine) and unsaturated synthetic lecithins (e.g., dialoylphosphatidylcholine or dilinoloyl-phosphatidylcholine) .
  • natural lecithins e.g., egg lecithin or soybean lecithin
  • synthetic lecithins such as saturated synthetic lecithins (
  • a steroid component can be incorporated into the lipid bilayer to increase the amount of water insoluble or sparingly- water soluble bioactive agents such as somatotropin which can be encapsulated by the liposomes
  • encapsulation is defined as th ' e entrapment of the bioactive agent within the aqueous compartment and/or within the membrane bilayer.
  • This enables the administration iji vivo of water-insoluble compounds, and it allows for the administration in vivo of high concentrations of water insoluble compounds or sparingly water-soluble compounds because it increases the dose:volume ratio of the compound.
  • the cholesterol also helps form a more closely packed bilayer system during preparation and inhibits the premature break ⁇ down of the liposomes by the intersticial and serum proteins of the host animal.
  • the steroids employed to practice this invention include, but are not limited to, cholesterol, coprostanol, cholestanol, cholestane, coprostane or epicholesterol and the like.
  • the liposomes used to practice the present invention can be prepared by any number of methods which are currently practiced in the art. Some examples of the methods that can be employed to prepare the liposomes include, but are not limited to, the methods of Bangham et al. (1965, J. Mol. Biol. 13:238- 252), Popahadjopoulos and Miller (1967, Biochem. Biophvs. Acta. 135:624-638), and Szoka and Popahadjopoulos in 1980, Ann. Rev. Biophvs. Bioen . , 9:467-508, and U.S. Patent Nos. 4,708,861, 4,235,871 and 4,891,208.
  • the preferred method involves preparing a methanol solution of phospholipids and mixing it with a solution of a steroid, such as cholesterol, in an organic solvent.
  • a steroid such as cholesterol
  • organic solvents are suitable, but halogenated hydrocarbons, diethyl ether or mixtures of halogenated hydrocarbons and diethyl ether are preferred.
  • Other organic solvents which can be employed as the solvent in the phospholipid or steroid mixture include, but are not limited to, ethanol, 2- propanol, isopropyl alcohol or combinations thereof.
  • the mole ratio of phospholipid to steroid is about 2:1 respectively, but it can range from as low as about 1:1 to as high as about 3:1.
  • the mixture is stirred to form a uniform solution and then the solvent is removed by evaporation.
  • Evaporation can be accomplished by any evaporative technique, e.g., by passing a stream of inert gas over the mixture, by heating, by vacuum or any combination thereof.
  • the dried lipid layer then is reconstituted with a neutral to slightly basic buffer solution (pH of about 7.0 to about 7.8) such as Tris buffer or HEPES buffer and the like.
  • the liposomes are formed by sonicating the solution in any appropriate sonicator. Sonicating the solution for about 5 minutes to about 15 minutes forms multila ellar vesicles (MLVs) while sonicating the solution for about 2 hours to about 4 hours forms unilamellar vesicles (UVs).
  • MLVs multila ellar vesicles
  • UVs unilamellar vesicles
  • aqueous solution containing from about 30 mg/ml to about 40 mg/ml of a polypeptide, such as somatotropin, from about 250 mg/ml to about 400 mg/ml of the chloride salt of an amino acid and from about 65 mg/ml to about 85 mg/ml of a carbohydrate is mixed with the reconstituted liposomes. It is not necessary to employ a carbohydrate in preparing the liposomes of the present invention. However, it is preferable to incorporate a carbohydrate into the liposomes since a carbohydrate can enhance the long-term stability of the liposomes.
  • Suitable carbohydrates include, but are not limited to, trehalose, sucrose, glucose, lactose, dextran and the like.
  • a preferred carbohydrate is trehalose.
  • the mixture then is either sonicated or microfluidized.
  • the mixture is microfluidized.
  • Microfluidization is a process which separates fluids into two phases and then allows the phases to rejoin to form lipophilic particles. Microfluidization is performed at from about 10,000 psi to about 20,000 psi for about 5 to about 15 cycles at room temperature (about 18°C to about 23°C).
  • the solution of liposome-encapsulated polypeptide then is added to a dilute aqueous mixture of a suspending agent (about a 1:3 ratio respectively) to disperse the liposome-encapsulated polypeptide to form a uniform suspension.
  • the dilute aqueous mixture of the suspending agent ranges from about 1% w/v to about 5% w/v of suspending agent to water.
  • the suspending agents employed to practice this invention include, but are not limited to, carbohydrates such as cellulosics, methylcellulose, starch and modified starch, agarose, gum arabic, ghatti, karay, tragacanth, guar, locust bean gum, tamarind, carrageenan, alginic acid, sodium alginate, xanthan, chickle, collagen, polyacrylamide, polysiloxanes, polyanhydrides, polyacrylates and amino acid polymers such as gelled albumin and other organic or inorganic polymers which can be mixed with liposomes in vitro.
  • the solution containing the liposome-encapsulated polypeptide is mixed with the suspending agent for about 2-5 minutes.
  • the resulting suspension can be divided up into desired individual volumes, lyophilized and then stored in a freezer for later use.
  • the amount of polypeptide encapsulated ranges from about 25% to about 45% of the mass of the lipids comprising the liposomes.
  • Encapsulated mass is the mass of the substance encapsulated per unit mass of the lipid. The term is commonly used to express the effectiveness of encapsulation and is expressed as a percentage.
  • Any polypeptide which stimulates an animal's immune system and can be incorporated into liposomes can be employed to practice this invention. Examples of preferred polypeptides include, but are not limited to, human somatotropin, bovine somatotropin, porcine somatotropin, ovine somatotropin, and the like.
  • somatotropins also can be employed, as well as any physiologically active fragment or analog thereof.
  • the preferred somatotropin used to practice this invention is a recombinant porcine somatotropin, such as delta-7 recombinant porcine somatotropin, described in European Patent Application Publication No. 104,920 (Biogen) , incorporated herein by reference.
  • growth factors include, but are not limited to, growth factors and any physiologically active fragments and analogs thereof.
  • Growth factors which can be employed to practice this invention include, but are not limited to, prolactin (PRL) , insulin-like growth factor I (IGF-I) and insulin like growth factor II (IGF-II), nerve growth factor (NGF), platelet- derived growth factor (PDGF), transforming growth factor- (TGF- ⁇ ), transforming growth factor- ⁇ (TGF- ⁇ ), fibroblast growth factor (FGF), epidermal growth factor (EGF), vaccinia growth factor (VGF) and the like.
  • Recombinant forms of the foregoing growth factors also can be employed as well as any physiologically active fragment thereof.
  • Preferred growth factors include recombinant human insulin-like growth factor I (rHuIGF-I) and recombinant human insulin-like growth factor II (rHuIGF-II).
  • the lyophilized liposome-encapsulated polypeptide can be reconstituted with sterile water for injection, bacteriostatic water for injection or any suitable aqueous preparation that can be safely administered parenterally to an animal.
  • routes of administration include, but are not limited to, inoculation or injection, (e.g., intraperitoneal, intramuscular, subcutaneous, intra-articular, intra- mammary, etc.).
  • a sufficient amount of the liposome- encapsulated polypeptide administered to an animal enhances the animal's immune system against many types of gram-negative bacteria such as various Salmonella species and Pasteurella species.
  • the amount of liposome-encapsulated polypeptide that can be administered to the host typically ranges from about 1 mg/kg to about 100 mg/kg of body weight. Preferably, from about 5 mg/kg of body weight to about 50 mg/kg of body weight is administered.
  • the liposome encapsulated somatotropin is especially effective in enhancing an animal's immune system against Salmonella typhimurium, Salmonella typhosa, Salmonella paratyphosa, and Pasteurella multocida .
  • a sufficient quantity of a solution of egg phosphatidylcholine (EPC) in methanol was mixed with a sufficient quantity of a solution of cholesterol in chloroform to form a uniform mixture of EPC/cholesterol having a molar ratio of 2:1 respectively.
  • the solution then was dried under nitrogen with vacuum desiccation for about twelve hours.
  • the dried lipid layer was reconstituted with a sufficient amount of 30mM Tris buffer to form a concentration of solute to solvent of 75 mg/ml.
  • the solution was sonicated for 2 hours in a Branson 5200 sonicator forming liposomes with unilamellar vesicles (UVs).
  • a solution containing 35 mg/ml zinc-complexed recombinant porcine somatotropin (ZnrpST) (Pitman-Moore Inc., lot pST D 148 010/011-
  • One-hundred thirty-two female hypophysectomized rats were employed to determine the effectiveness of somatotropin in enhancing host protection against Salmonella typhimurium.
  • the intact rats were obtained from Holtzman Company, Madison, Wisconsin, and they were hypophysectomized at Johnson Laboratories, Bridgeview, Illinois, before they were delivered to Pitman-Moore, Inc., Terre Haute, Indiana, for study.
  • the rats were divided into five groups, each group comprising at least twelve rats. Each group was treated with a different preparation.
  • the first group of thirty-six rats was treated with a placebo composed of Parlow's Buffer at a dose of 200 ⁇ l/rat/day (negative control); the second group of thirty-six rats was treated with recombinant rat interferon gamma (rRalFN-c) at a dose of 500 U/rat/day with a specific activity of 4xl0 6 U/mg of protein (positive control) (obtained from AMGEN, La Jolla, CA) ; the third group of twenty-four rats was treated with pituitary-derived porcine somatotropin (npST) at a dose of 48 ⁇ g/rat/day (obtained from A.F.
  • rRalFN-c recombinant rat interferon gamma
  • positive control obtained from AMGEN, La Jolla, CA
  • npST pituitary-derived porcine somatotropin
  • each rat was infected intraperitoneally (i.p.) with an infectious dose of Salmonella typhimurium, 7.5xl0 7 viable colony forming units (CFU), (strain 84-4728, supplied by B.O. Blackburn, U.S.D.A. Infectious Disease Center, Ames IA) .
  • CFU colony forming units
  • Treatment regimens were continued for an additional six days after i.p. administration of the infectious bacteria.
  • Figure 1 is a plot of the data from Table I, and more clearly illustrates the increased survival rates of rats treated with the rRalFN-c and somatotropin versus the placebo. TABLE I Percent Survival
  • Example 2 Laboratories, Bridgeview, Illinois) according to the protocol outlined in Example 2. Fifteen rats were treated with placebo at a dose of 200 ⁇ l/rat/day; fifteen rats were treated with rRalFN-c at a dose of
  • the liposome-encapsulated rpST was prepared according to the method disclosed in Example 1. The free liposomes were prepared by the same method as the liposomes in Example 1 except rpST was not employed in the preparation.
  • Each rat was infected intraperitoneally (i.p.) with an infectious dose of Salmonella typhimurium (6.7xl0 7 CFU). After infection with the bacteria, each rat was treated subcutaneously (s.c.) with the respective preparation for six consecutive days. Survivability of the hypophysectomized rats was followed for 14 days. Table III discloses the survivability of the hypophysectomized rats treated with the five preparations. All rats which were treated with placebo died by the seventh day (0/12, 0%) whereas rpST (8/12, 67%), rpST/liposomes (10/12, 83%) and rRalFN-c (8/12, 67%) all significantly increased (P ⁇ 0.05) host survival rates.

Abstract

Procédé de renforcement de la protection contre un agent pathogène chez un hôte par l'encapsulement d'un polypeptide dans des liposomes et par l'administration à un animal d'un quantité suffisante de polypeptides de ce type pour que ledit polypeptide renforce la capacité des cellules phagocytaires du système immunitaire de l'animal à tuer un microorganisme pathogène.
PCT/US1992/007562 1991-09-11 1992-09-04 Procede de renforcement du systeme immunitaire d'un hote a l'aide de polypeptides encapsules dans des liposomes WO1993004672A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US75760191A 1991-09-11 1991-09-11
US757,601 1991-09-11

Publications (1)

Publication Number Publication Date
WO1993004672A1 true WO1993004672A1 (fr) 1993-03-18

Family

ID=25048473

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1992/007562 WO1993004672A1 (fr) 1991-09-11 1992-09-04 Procede de renforcement du systeme immunitaire d'un hote a l'aide de polypeptides encapsules dans des liposomes

Country Status (2)

Country Link
AU (1) AU2583892A (fr)
WO (1) WO1993004672A1 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2070774A1 (es) * 1993-09-16 1995-06-01 Hipra Lab Sa Preparaciones liposomicas de farmacos o antigenos vacunales para medicina veterinaria.
EP0664129A2 (fr) * 1994-01-25 1995-07-26 PHARMACIA S.p.A. Epicholesterol pour le traitement des dyslipidémies
WO2000044758A1 (fr) * 1999-02-01 2000-08-03 Eisai Co., Ltd. Compose d'adjuvant immunologique
US6521776B2 (en) 2000-07-31 2003-02-18 Eisai Company, Limited Immunological adjuvant compounds compositions and methods of use thereof
US7560584B2 (en) 1999-02-01 2009-07-14 Eisai R&D Management Co., Ltd. Immunomodulatory compounds and methods of use thereof
US7833993B2 (en) 1999-02-01 2010-11-16 Eisai R&D Management Co., Ltd. Immunomodulatory compounds and methods of use thereof
US7915238B2 (en) 1999-02-01 2011-03-29 Eisai R & D Management Co., Ltd. Immunomodulatory compounds and methods of use thereof
US7976852B2 (en) 2005-04-26 2011-07-12 Eisai R&D Management Co., Ltd. Compositions and methods for cancer immunotherapy
CN101213199B (zh) * 2005-06-30 2013-12-18 卫材R&D管理有限公司 用于制备免疫佐剂的化合物
CN104602672A (zh) * 2012-06-14 2015-05-06 伯尔尼大学 用于治疗细菌感染的定制脂质体

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1985003640A1 (fr) * 1984-02-15 1985-08-29 The Liposome Company, Inc. Compositions de gels et de liposomes
EP0308197A2 (fr) * 1987-09-14 1989-03-22 Pitman-Moore, Inc. Méthode pour stimuler le système immunitaire
WO1989005151A1 (fr) * 1987-12-04 1989-06-15 The Liposome Company, Inc. Liposomes a degre de stabilite eleve et procede de preparation et d'utilisation de tels liposomes
WO1990003795A1 (fr) * 1988-10-05 1990-04-19 Vestar, Inc. Procede de fabrication de liposomes a stabilite amelioree pendant le sechage
EP0393707A2 (fr) * 1989-04-21 1990-10-24 Otsuka Pharmaceutical Co., Ltd. Composés bio-actifs associés à des liposomes et leur utilisation dans des préparations pharmaceutiques
WO1991001719A1 (fr) * 1989-08-01 1991-02-21 The University Of Michigan Administration topique de peptides/proteines enfermes dans des liposomes deshidrates/rehydrates

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1985003640A1 (fr) * 1984-02-15 1985-08-29 The Liposome Company, Inc. Compositions de gels et de liposomes
EP0308197A2 (fr) * 1987-09-14 1989-03-22 Pitman-Moore, Inc. Méthode pour stimuler le système immunitaire
WO1989005151A1 (fr) * 1987-12-04 1989-06-15 The Liposome Company, Inc. Liposomes a degre de stabilite eleve et procede de preparation et d'utilisation de tels liposomes
WO1990003795A1 (fr) * 1988-10-05 1990-04-19 Vestar, Inc. Procede de fabrication de liposomes a stabilite amelioree pendant le sechage
EP0393707A2 (fr) * 1989-04-21 1990-10-24 Otsuka Pharmaceutical Co., Ltd. Composés bio-actifs associés à des liposomes et leur utilisation dans des préparations pharmaceutiques
WO1991001719A1 (fr) * 1989-08-01 1991-02-21 The University Of Michigan Administration topique de peptides/proteines enfermes dans des liposomes deshidrates/rehydrates

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2070774A1 (es) * 1993-09-16 1995-06-01 Hipra Lab Sa Preparaciones liposomicas de farmacos o antigenos vacunales para medicina veterinaria.
EP0664129A2 (fr) * 1994-01-25 1995-07-26 PHARMACIA S.p.A. Epicholesterol pour le traitement des dyslipidémies
EP0664129A3 (fr) * 1994-01-25 1997-08-20 Pharmacia Spa Epicholesterol pour le traitement des dyslipidémies.
US7833993B2 (en) 1999-02-01 2010-11-16 Eisai R&D Management Co., Ltd. Immunomodulatory compounds and methods of use thereof
US6290973B1 (en) 1999-02-01 2001-09-18 Eisai Co., Ltd. Immunological adjuvant compounds, compositions, and methods of use thereof
EP1439184A1 (fr) * 1999-02-01 2004-07-21 Eisai Co., Ltd. Composés adjuvants immunologiques
US7560584B2 (en) 1999-02-01 2009-07-14 Eisai R&D Management Co., Ltd. Immunomodulatory compounds and methods of use thereof
WO2000044758A1 (fr) * 1999-02-01 2000-08-03 Eisai Co., Ltd. Compose d'adjuvant immunologique
US7915238B2 (en) 1999-02-01 2011-03-29 Eisai R & D Management Co., Ltd. Immunomodulatory compounds and methods of use thereof
US6521776B2 (en) 2000-07-31 2003-02-18 Eisai Company, Limited Immunological adjuvant compounds compositions and methods of use thereof
US7976852B2 (en) 2005-04-26 2011-07-12 Eisai R&D Management Co., Ltd. Compositions and methods for cancer immunotherapy
US8603482B2 (en) 2005-04-26 2013-12-10 Eisai R&D Management Co., Ltd. Compositions and methods for cancer immunotherapy
CN101213199B (zh) * 2005-06-30 2013-12-18 卫材R&D管理有限公司 用于制备免疫佐剂的化合物
CN104602672A (zh) * 2012-06-14 2015-05-06 伯尔尼大学 用于治疗细菌感染的定制脂质体
CN104602672B (zh) * 2012-06-14 2018-12-25 伯尔尼大学 用于治疗细菌感染的定制脂质体
EP2861214B1 (fr) * 2012-06-14 2020-08-05 Universität Bern Liposomes personnalisés pour le traitement des infections bactériennes
US10744089B2 (en) 2012-06-14 2020-08-18 Universitaet Bern Tailored liposomes for the treatment of bacterial infections

Also Published As

Publication number Publication date
AU2583892A (en) 1993-04-05

Similar Documents

Publication Publication Date Title
US6455088B1 (en) Peptide/lipid complex formation by co-lyophilization
US4863740A (en) Interleukin therapy
US4241046A (en) Method of encapsulating biologically active materials in lipid vesicles
CA2515892C (fr) Vehicule d'administration de medicament lipophile et methodes d'utilisation correspondantes
US4235871A (en) Method of encapsulating biologically active materials in lipid vesicles
EP0937456B1 (fr) Dispersion liposomale d'érythropoietine
WO1993004672A1 (fr) Procede de renforcement du systeme immunitaire d'un hote a l'aide de polypeptides encapsules dans des liposomes
WO1991004019A1 (fr) Peptides et proteines therapeutiques
Bergers et al. Interleukin-2-containing liposomes: interaction of interleukin-2 with liposomal bilayers and preliminary studies on application in cancer vaccines
EP1064950A1 (fr) Nouveaux remedes contre l'hepatite
JPH06345663A (ja) バンコマイシン含有リポソーム製剤
JPH08505403A (ja) 安定なタンパク質：リン脂質組成物及び方法
JP2817883B2 (ja) 高度に完全なリポソームおよびその製剤法と用途
JP2660341B2 (ja) リポソーム製剤およびその製造法
MXPA00003132A (en) Peptide/lipid complex formation by co-lyophilization

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AT AU BB BG BR CA CH CS DE DK ES FI GB HU JP KP KR LK LU MG MN MW NL NO PL RO RU SD SE

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL SE BF BJ CF CG CI CM GA GN ML MR SN TD TG

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

NENP Non-entry into the national phase

Ref country code: CA

122 Ep: pct application non-entry in european phase