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WO1994027718A1 - Preparation de microparticules et procede d'immunisation - Google Patents

Preparation de microparticules et procede d'immunisation Download PDF

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Publication number
WO1994027718A1
WO1994027718A1 PCT/US1994/005834 US9405834W WO9427718A1 WO 1994027718 A1 WO1994027718 A1 WO 1994027718A1 US 9405834 W US9405834 W US 9405834W WO 9427718 A1 WO9427718 A1 WO 9427718A1
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WO
WIPO (PCT)
Prior art keywords
microparticles
pharmaceutical composition
polymer
antigen
mammal
Prior art date
Application number
PCT/US1994/005834
Other languages
English (en)
Inventor
Derek Thomas O'hagan
John Paul Mcgee
Stanley Stewart Davis
Original Assignee
Hagan Derek Thomas O
John Paul Mcgee
Stanley Stewart Davis
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 Hagan Derek Thomas O, John Paul Mcgee, Stanley Stewart Davis filed Critical Hagan Derek Thomas O
Priority to US08/374,751 priority Critical patent/US5603960A/en
Priority to AU70441/94A priority patent/AU7044194A/en
Publication of WO1994027718A1 publication Critical patent/WO1994027718A1/fr

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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
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/5005—Wall or coating material
    • A61K9/5021—Organic macromolecular compounds
    • A61K9/5031—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)
    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
    • A61K9/1647—Polyesters, e.g. poly(lactide-co-glycolide)
    • 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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1682—Processes
    • A61K9/1694—Processes resulting in granules or microspheres of the matrix type containing more than 5% of excipient

Definitions

  • the present invention relates to a method for producing microparticles useful in the formulation of pharmaceutical compositions.
  • the present invention further relates to a method of immunizing a mammal against diseases comprising administering to a mammal an effective amount of antigen containing microparticles.
  • the present invention describes a method of potentiating an immune response in a mammal comprising administering an effective amount of a pharmaceutical composition containing said microparticles to a mammal.
  • the present invention further describes a vaccine comprising a pharmaceutical composition containing said microparticles.
  • An antigen delivery system comprising microparticles containing entrapped antigens is further described by the present invention.
  • a pharmaceutical composition comprising microparticles and a pharmaceutical carrier is also provided.
  • Biodegradable polymers such as polylactide-co- glycolides (PLG) have been used to encapsulate proteins
  • microcapsules •*-5 in microcapsules to deliver "pulses” (i.e. "intermittent doses") of antigenic material for the development of vaccines (see e.g. United States Patent No. 5,075,109 to Tice et al. ) .
  • the use of microencapsulation to protect sensitive bioactive agents against degradation is well
  • the drug release pattern for a microcapsule is dependent upon numerous factors. For example, the type
  • 25 of drug encapsulated and the form in which it is present may affect the drugs release pattern.
  • Another factor which may affect the drug release pattern is the type of polymer used to encapsulate the drug.
  • release pattern include the drug loading, the manner of distribution in the polymer, the particle size and the particle shape.
  • microparticles There are several methods known for the production of microparticles. Typical methods for producing microparticles include solvent evaporation and phase separation. With production methods such as solvent evaporation, as much as 50% w/w of insoluble or poorly soluble materials, may be incorporated in biodegradable microparticles. However, with more water soluble materials, such as peptides, drug loadings have generally been much lower.
  • phase separation methods of microparticle preparation allow a more efficient incorporation of drugs and can easily be scaled up for industrial purposes.
  • the process of phase separation usually employs an emulsion or a suspension of the drug particles in a solution of a high molecular weight polymer and an organic polymer solvent.
  • a non-solvent is then added to the suspension or emulsion, causing the polymer to separate from solution and to encapsulate the suspended drug particles or droplets containing them.
  • the resulting microparticles (which are still swollen with solvent) are then normally hardened by a further addition of a non-solvent or by some other process which strengthens and improves the properties of the microparticles.
  • Such problems include: low or negligible and inefficient drug entrapment ( ⁇ 0.5% w.w), aggregation of particles, formation of non-spherical particles, formation of particles with surfaces that are not smooth and which have defects, the presence of large particles with a wide range of sizes (5 ⁇ m-250 ⁇ m) and the presence of non-particulate material. All these problems reduce the effectiveness and reproducability of the microparticles produced by these methods for use in controlled release delivery systems.
  • the present invention solves many of the problems associated with current immunization methods.
  • the present invention provides an essentially continuous release of an antigen from microparticles prepared using the novel method described by the present invention. It has been surprisingly discovered in accordance with the present invention that a continuous release of antigen results in the induction of immune responses which are comparable to those induced by the potent immunological adjuvant, aluminum hydroxide.
  • the present invention provides a method for producing microparticles useful in the formation of pharmaceutical compositions.
  • the average microparticle size is between 200 nm to 200 ⁇ m.
  • the present invention further provides a method of immunizing a mammal against diseases comprising administering to a mammal an effective amount of antigen containing microparticles.
  • the microparticles are administered orally or parenterally.
  • Another aspect of this invention is directed to a method of potentially an immune response.
  • Yet another embodiment of the present invention provides an antigen delivery system comprising microparticles containing entrapped antigens.
  • Still a further aspect of the present invention provides a pharmaceutical composition comprising microparticles and a pharmaceutically acceptable carrier.
  • a further aspect of the present invention provides a vaccine comprising a pharmaceutical composition containing said microparticles.
  • Fig. 1 shows an essentially continuous release of entrapped ovalbumin (OVA) in microparticles prepared with polymer RG 503 over 10 to 15 days ij vitro.
  • OVA ovalbumin
  • Fig. 2 shows an essentially continuous release of entrapped ovalbumin (OVA) in microparticles prepared with polymer R 208 over 40 days iji vitro.
  • OVA ovalbumin
  • Fig. 3 shows that the serum IgG antibody response to OVA in microparticles (OVA/PLG) and the response to OVA absorbed to Alum (OVA/Alum) were significantly enhanced in comparison to the response to soluble OVA for mice parenterally immunized.
  • Fig. 4 shows that the serum IgG antibody response to OVA in microparticles (OVA/PLG) was significantly enhanced in comparison to the response to soluble OVA for orally immunized mice.
  • the present invention is directed to a method for the production of microparticles useful in the formulation of a pharmaceutical composition.
  • the first medium is a non-solvent of a pharmacologically-acceptable polymer containing an aqueous solution of the bioactive material to be encapsulated (e.g. an aqueous solution of an antigen).
  • the second medium is a solvent containing a pharmacologically-acceptable polymer dissolved in the solvent.
  • the second medium is added to first medium, causing the polymer to precipitate from the solution and to microencapsulate the bioactive material as it separates, forming microparticles. Additional treatment of the microparticles such as further hardening or washing can then be carried out as appropriate.
  • the process of the present invention is distinguished from those in the prior art by the use of a phase-inducing agent for the formation of the dispersion of the material to be microencapsulated.
  • This variation from the prior art leads to a process which provides microparticles of particular value.
  • the material to be encapsulated by way of the novel process may be coated with a single wall or "shell" of polymeric material (microcapsules) or may be homogeneously dispersed within a polymeric matrix (microspheres) .
  • microparticles includes both microcapsules and microspheres and the term microencapsulation or encapsulation should be construed accordingly.
  • the novel process may be used to encapsulate a variety of materials.
  • the bioactive materials that may be encapsulated in microparticles include agricultural agents such as insecticides, fungicides, herbicides, rodenticides, pesticides, fertilizers and viruses for crop protection, as well as cosmetic agents such as deodorants and fragrances, and food additives such as flavors.
  • the microparticles of the present invention are used with pharmaceutical (bioactive) agents for prophylactic, therapeutic or even diagnostic use.
  • the preferred pharmaceutical agents of the present invention are immunogens and drugs, especially those of a water-soluble nature. Additional preferred pharmaceutical agents include enzymes, steroids, hormones, and proteins or peptides.
  • the most preferred pharmaceutical agents of the present invention are proteins or peptides which are antigens or portions thereof that are designed to induce an immunogenic response.
  • the pharmaceutical agents which are recombinant proteins or synthetic peptides are microencapsulated according to the method described by the present invention.
  • a pharmacologically acceptable polymer is biocompatible as well as biodegradable (i.e. the polymer is substantially non- toxic to the host and of such composition that it is degradable by the body into metabolic products that have no substantial deleterious or untoward effects on the body) .
  • biodegradable polymer is biocompatible as well as biodegradable (i.e. the polymer is substantially non- toxic to the host and of such composition that it is degradable by the body into metabolic products that have no substantial deleterious or untoward effects on the body) .
  • biodegradable i.e. the polymer is substantially non- toxic to the host and of such composition that it is degradable by the body into metabolic products that have no substantial deleterious or untoward effects on the body
  • alpha-hydroxy-carboxylic acids and certain lactones can be condensed to form such polymers, particularly lactic acid and glycolic acids, or combinations thereof (see, for example. United States Patent 3,773,919 to Boswell et al. ) .
  • Similar biocompatible polymers based on glycolic acid and glycerol and the like are known (see, for example. United States Patents 3,991,776 to Schmitt, et al. and 4,076,779 and 4,188,470 to Casey et al. ) .
  • the pharmacologically acceptable polymer preferably used for encapsulating the bioactive material of the present invention is a polylactide polymer (PLA), or particularly a polylactide-co-glycolide polymer (PLG) .
  • PLA polylactide polymer
  • PLAG polylactide-co-glycolide polymer
  • the ratio of lactide to glycolide in the most preferred pharmacologically acceptable polymer ultimately determines the rate of release of the bioactive material from the microcapsules, and can thus be varied, depending on the desired mode of delivery of the microparticles and the contents thereof.
  • the molar ratio of lactide to glycolide will be between 100:0 and 0:100. In a more preferred embodiment, the molar ratio of lactide to glycolide will be preferably between 70:30 and 30:70.
  • a preferred PLG polymer has a lactide:glycolide ratio of 50:50 and a molecular weight of 9,000 although other polymers which have been used are a PLG polymer having a lactide:glycolide ratio of 85:15 and a molecular weight of 54,000 and a PLA polymer with a molecular weight of 300,00. It is possible to administer microparticles made from more than one biodegradable polymer or made from different ratios of the same polymer. By utilizing a combination of various polymers with different lactide/glycolide ratios, the release profile of the encapsulated agent can be controlled.
  • PLG polymers undergo biodegradation by random, non-enzymatic scission to form the endogenous metabolites lactic acid and glycolic acid.
  • PLG microparticles release entrapped pharmaceutical agents as a function of time, by one or more mechanisms, but the release is mainly controlled by bulk degradation of the polymer.
  • microparticles can be prepared that release their agent over a period of days to in excess of 1 year.
  • Mixed populations of PLG microparticles prepared from different polymeric compositions and molecular weights may be engineered to create an essentially continuous release of bioactive materials at predetermined intervals. For vaccination purposes, this would obviate the need for booster injections.
  • an essentially continuous release describes the rate of release of the bioactive material from the microparticle into a mammal necessary to provide the required immune response to treat the requisite disease.
  • Microencapsulation can also be used to slow the release of a drug in the body. This has advantages in that a single essentially continuous release dose may replace several separate doses of a non-encapsulated drug. This may decrease the toxic side effects of some drugs by avoiding the high initial concentrations of drug in the blood, which often occurs following conventional administration. In some cases, it may be desirable to have an essentially continuous release pattern with the microparticles delivering a fixed amount of drug per minute, hour or day during the period of their effectiveness.
  • the first medium is preferably selected from oils such as silicone oils, mineral oils, petroleum oils, sesame oil, peanut oil, soybean oil, corn oil, cotton seed oil, coconut oil and linseed oil.
  • the first medium is a silicone oil.
  • the second medium is preferably an organic solvent such as chloroform, methylene chloride, ethylene chloride, ethylene dichloride, ethyl acetate, methylchloroform, tetrahydrofuran or benzene.
  • methylene chloride (dichloromethane) and in particular acetate are the second medium, especially when a PLG polymer is used.
  • the ratio by volume of the solution of polymer in the second medium, during the dispersion (as is commonly the case) of material in the first medium it has been found that the preferred ratio lies in the range of 1:5.2 to 1:4.8.
  • a ratio of 1:5.0 is required in order to produce microparticles when using a polymer concentration of about 2% w/v. Below this range, the microparticles are less uniform; while above this range, there is an increasing tendency for matrix formation.
  • the preferred ratio will move towards a smaller proportion of the polymer solution and vice versa for a polymer concentration higher than 2% w/v.
  • the preferred range of ratios is 1:3.75 to 1:3.0.
  • the range of ratios in the present embodiment is 1:2.72 to 1:2.3.
  • the use of larger volumes of more concentrated polymer solution may enable a higher degree of entrapment of the material to be achieved.
  • the temperature at which the process is carried out it is often preferable that this is 25°C or less, if the material to be microencapsulated is temperature sensitive, which can often be the case. However, it is preferable that the temperature is kept within a range of about 10° to 25°C, for example at 12°C or 22°C, since below this range, the increased viscosity of the first medium can deleteriously affect microparticle formation. It is also desirable to maintain the temperature well below the boiling point of the second medium, which for dichloromethane, for example, is 40°C.
  • the process is enhanced through the use of a surfactant, preferably one which is non-ionic such as a sorbitan ester, for example Span 40. The preferred amount of surfactant equal to about 15% by weight of the polymer.
  • the formed microparticles are further hardened in a third medium, which is desirably a non-solvent for the polymer.
  • a third medium may be an alkane or halogenated alkane or a volatile silicone oil.
  • the third medium of the preferred embodiment is heptane.
  • the superior surface morphology which may be achieved with the microparticles of the invention may be determined by the measurement of the rugosity of the particles.
  • the rugosity values are those measured by air permeametry. Measurement of the rugosity by air permeametry produces a value which reflects the nature of the external surface of the material under test. The lower the rugosity value, the smoother the external surface of the microparticles.
  • the invention includes particles having a rugosity value of less than 2.0. In practice, the smoothness of the novel microparticles is readily apparent under the scanning electron microscope wherein the lack of larger particulate material may also be observed.
  • the average size of the microparticles produced by the novel process is between 200 nm to 200 ⁇ m characterized in that at least 90% by weight of said particles have a size which is within ⁇ 10% of the mean particle size.
  • a pharmaceutical composition containing the microparticles of the present invention is administered orally or parenterally.
  • the .microparticles are preferably between 100 nm to 10 ⁇ m in size.
  • the microparticles may be larger, preferably between 5 ⁇ m and 200 ⁇ m and especially between 10 ⁇ m and 100 ⁇ m.
  • Parenteral administration may be by any of the normal routes, for example, intravenously, intramuscularly, intraperitoneally but is more preferably by subcutaneous injection.
  • the present invention further describes a composition for oral or parenteral administration comprising microparticles wherein the average size is in a range as indicated above and especially in which at least 90% by weight. In a preferred embodiment, at least 95% by weight of the microparticles have a size falling within the desired range.
  • the composition described by the present invention may also contain a pharmaceutically acceptable carrier.
  • a pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and the like. The use of such media and agents is well-known in the art.
  • the amount of pharmaceutical agent incorporated in the microparticles depends upon the starting amount of material used. Thus, it has been found that higher levels of entrapment are obtained with higher starting levels of material.
  • the particles contain up to 20% w/w of drug loading, conveniently between 0.01% w/w and 10% by weight of material. This amount will vary in particular with the desired dosage of a pharmaceutical agent.
  • microparticles produced by the novel process of the present invention may be formulated into various forms of composition depending upon the nature of the material contained therein.
  • the microparticles when they encapsulate a pharmaceutical agent, they may be formulated into a pharmaceutical composition together with a physiologically acceptable diluent or carrier for administration. They may be administered by any means or route desired. In the case of the administration of pharmaceuticals to a patient this may be oral administration or preferably parenteral administration, especially by injection which is most preferably intramuscular or especially subcutaneous.
  • the composition may particularly be adapted for either oral or parenteral administration to a patient.
  • the microparticles When administered parenterally, for example subcutaneously, the microparticles are preferably suspended in a pharmaceutically acceptable carrier which is sterile and pyrogen free. When administered orally, the microparticles are preferably mixed with a pharmaceutically acceptable carrier which is a solid. If the microparticles are to be administered by injection they may first be suspended in a pharmaceutically acceptable carrier. If the pharmaceutical composition is a vaccine, an adjuvant such as aluminum hydroxide may be used. The exact nature of the composition will depend upon the amount of agent to be administered, the suspending capacity of the pharmaceutically acceptable carrier and the volume of solution which can be injected at a particular site or i n a particular subject.
  • the present invention is further directed to a method of immunizing a mammal against disease comprising administering to a mammal an effective amount of antigen containing microparticles.
  • the present invention describes a method of potentiating an immune response in a mammal comprising administering an effective amount of a pharmaceutical composition containing said microparticles to a mammal.
  • an effective amount of pharmaceutical composition is the amount of composition necessary to treat the particular disease being treated.
  • the microparticles within the composition are produced as previously described.
  • the pharmaceutical composition may be administered orally or parenterally using conventional techniques previously described.
  • the present invention is also directed to an antigen delivery system comprising the microparticles described by the present invention containing an antigenic material.
  • antigenic material can include but is not limited to the desired antigen peptide, any peptides produced during the synthesis of the desired antigenic peptide, a combination of several desired peptides or the peptides produced during the synthesis of the antigenic peptides and peptides chemically linked to lipids.
  • the present invention also describes a vaccine comprising a pharmaceutical composition containing said microparticles.
  • the vaccines can be administered by any of several routes including parenterally or orally in a single dose.
  • the dose of bioactive material ranges from about 1 ⁇ g to about 500 ⁇ g.
  • the dosage of bioactive material ranges from 1 ⁇ g to 10 mg.
  • the vaccines of the present invention are administered to mammals.
  • the vaccine can be formulated with any other pharmaceutically acceptable carrier.
  • OVA antigen
  • silicone oil Dow Corning 200/1000
  • PLG polylactide-co-glycolide
  • dichloromethane 2% w/v
  • the mixture was then transferred to 300 ml of heptane and stirred for 30 minutes.
  • the heptane was decanted and an additional 300 ml of heptane was added.
  • the mixture was stirred for an additional 30 minutes and then decanted.
  • the microparticles were then washed twice with 50 ml of water and recovered by centrifugation.
  • Example 2 20 to 30 mg of dry microparticles prepared by the first method described in Example 1 were dissolved in dichloromethane and the OVA was extracted. The OVA content was then determined using a bicinchoninic acid (BCA) protein assay method. The samples were viewed under a scanning electron microscope (s.e.m.) to determine: a. if the particles were smooth, spherical, discrete and free from surface defects; b. whether or not there was any non-particulate material present; c. to estimate the size and polydispersity of the particles.
  • BCA bicinchoninic acid
  • the size of the microparticles was confirmed by laser diffractometry using a Malvern Laser sizer 2000D. Samples were analyzed using secondary ion mass spectrometry (SIMS): a. to determine if the OVA was present on the surface of the particles and/or if the OVA was entrapped by the polymer; b. to show if there was any residual silicone oil in the particles. A Western blot was carried out to determine whether or not antibodies raised to OVA still recognized OVA when run on a gel.
  • SIMS secondary ion mass spectrometry
  • microparticles Three batches of microparticles were prepared with polymer RG 503 containing 1.2, 2.7 and 5.1% w/w entrapped OVA and three batches were "prepared with polymer R 208 containing 2.3, 3.9 and 6.5% w/w entrapped OVA.
  • the rates of release of protein from the microparticles were determined ijri vitro.
  • a known weight of microparticles (30 mg) was placed into a number of glass vials in 10 ml phosphate buffered saline and the vials were placed in a shaking water bath at 37°C. At selected intervals, one vial was removed and the buffer was filtered through a 0.2 mm filter into a clean vial and freeze dried.
  • the levels of released OVA in the samples were assayed using a Bicinchoninic protein assay after reconstitution of the vials.
  • Figure 1 shows the microparticles prepared with RG 505 using the novel phase separation technique showed a slow and steady release of entrapped OVA over 10 to 15 days in vitro.
  • the microparticles prepared with R 208 using the novel phase separation technique showed a slow and steady release of entrapped OVA over 40 days in vitro as shown in Figure 2.
  • the rate of release was dependent on the level of loading and the microparticles with higher loading levels released the entrapped OVA more quickly.
  • mice Three groups of ten female BALB/c mice were each immunized subcutaneously with lOO ⁇ g OVA either entrapped in microparticles, adsorbed to a 2% suspension of alum (Alu-Gel-S, Serva, Heidelberg) or dissolved in saline. Immediately before administration the required dose of freeze dried microparticles was suspended in physiological saline. Identical booster doses were administered to each study group six weeks after the primary immunization. Blood samples were collected from the tail veins of the mice at two week intervals for twelve weeks, then every four weeks.
  • mice Two groups of ten female BALB/c mice each received primary immunization with 1 mg OVA by gastric intubation on three consecutive days, either as soluble antigen, or entrapped in microparticles. Immediately before administration, the required dose of microparticles was resuspended in phosphate buffered saline. Four weeks after the primary immunizations, the two groups of animals were reimmunized with the same dose of OVA in the form previously administered. Blood samples were collected from the tail veins every two weeks.
  • each serum sample was determined in an established ELISA as previously described O'Hagan in 1991 Immunology 7_3 :239 ⁇ 242 and Vaccine £:768-771 and was standardized against a positive control anti-serum obtained by hyperimmunization of a mouse with OVA in Freund's complete adjuvant.
  • Each serum sample from each mouse was assayed at four different dilutions. The results are expressed as mean antibody units for the groups of mice, calculated from the standard curve obtained from the hyperimmune mouse serum diluted between 1/500 and 1/64,000. The value for each dilution fell on the standard curve and its value was taken as the mean of the four separate dilutions.
  • Figure 3 shows that following booster immunizations at week six, the serum IgG antibody response to OVA in microparticles and the response to OVA adsorbed to Alum were significantly enhanced in comparison to the response to soluble OVA for mice parenterally immunized.
  • Figure 4 shows that following booster immunizations at week four, the serum IgG antibody response to OVA in microparticles was significantly enhanced in comparison to the response to soluble OVA for orally immunized mice. Furthermore, following parenteral immunization, the highest antibody response (325 antibody units) was obtained at week 10 in the group administered microparticles. Thus, the present invention shows that oral immunization with microparticles induced high levels of serum antibodies that were about half the optimal levels induced by primary and secondary parenteral immunization with microparticles or alum.
  • microparticle characteristics may be manipulated to minimize the burst effect, it is clear that controlled release systems prepared from PLG and related polymers normally show a release profile incorporating a substantial burst. Therefore, it is encouraging that the novel microparticle preparation technique described in the present invention produces microparticles which display an essentially continuous x release. As an illustration of this, the same protein was entrapped in microparticles prepared from the same polymer (RG 503), but prepared by a solvent evaporation technique.
  • Alum-adjuvanted preparation as shown in Figure 3.
  • a systemic IgG antibody response significantly greater than the response to soluble OVA was observed following a boost 0 at four weeks as shown in Figure 4.
  • the antibody level for the microparticle group was comparable to the soluble OVA group.
  • microparticles prepared by the novel method as described in the present invention exhibited continuous release of antigen, but also induced enhanced antibody responses.
  • release profile displayed by the model protein from the microparticles in the present study would appear to be a potentially attractive release profile for a range of macromolecular drugs that normally require frequent injections.

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Abstract

L'invention concerne un procédé de production de microparticules utiles dans la formulation de compositions pharmaceutiques. L'invention concerne également un procédé d'immunisation d'un mammifère contre des maladies, consistant à administrer à ce mammifère une dose efficace de microparticules contenant des antigènes. L'invention a notamment trait à un procédé de potentialisation d'une réponse immunitaire chez un mammifère, consistant à administrer une dose efficace d'une composition pharmaceutique à ce mammifère. De plus, l'invention a trait à un vaccin comprenant une composition pharmaceutique contenant lesdites microparticules. Un système d'apport d'antigènes comprenant des microparticules contenant des antigènes piégés, ainsi qu'une composition pharmaceutique comprenant des microparticules ainsi qu'un excipient pharmaceutique sont également traités dans ladite invention.
PCT/US1994/005834 1993-05-25 1994-05-24 Preparation de microparticules et procede d'immunisation WO1994027718A1 (fr)

Priority Applications (2)

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US08/374,751 US5603960A (en) 1993-05-25 1994-05-24 Preparation of microparticles and method of immunization
AU70441/94A AU7044194A (en) 1993-05-25 1994-05-24 Preparation of microparticles and method of immunization

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GB9310781.1 1993-05-25
GB939310781A GB9310781D0 (en) 1993-05-25 1993-05-25 Preparation of microparticles

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0737472A1 (fr) * 1995-03-16 1996-10-16 LG Chemical Limited Formulation de vaccin pour une administration unique
WO1998052605A1 (fr) * 1997-05-19 1998-11-26 Sumitomo Pharmaceuticals Company, Limited Composition immunostimulante
WO1999009956A1 (fr) * 1997-08-29 1999-03-04 Corixa Corporation Agents bioactifs encapsules a liberation rapide servant a provoquer ou a potentialiser une reponse immunitaire, et procedes d'utilisation de ceux-ci
WO1999047588A1 (fr) * 1998-03-14 1999-09-23 Cenes Drug Delivery Limited Production de microparticules
WO2003080033A1 (fr) * 2002-03-21 2003-10-02 Jagotec Ag Microparticules
US7285539B2 (en) 1999-10-27 2007-10-23 Chiron Corporation Activation of HCV-specific T cells
US7501134B2 (en) * 2002-02-20 2009-03-10 Novartis Vaccines And Diagnostics, Inc. Microparticles with adsorbed polypeptide-containing molecules
EP1579851A3 (fr) * 1997-08-29 2009-09-02 Corixa Corporation Agents bioactifs encapsulés à libération rapide servant à initier ou à potentialiser une réponse immunitaire, et procédés d'utilisation de ceux-ci

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EP0737472A1 (fr) * 1995-03-16 1996-10-16 LG Chemical Limited Formulation de vaccin pour une administration unique
US5753234A (en) * 1995-03-16 1998-05-19 Lg Chemical Ltd. Single-shot vaccine formulation
WO1998052605A1 (fr) * 1997-05-19 1998-11-26 Sumitomo Pharmaceuticals Company, Limited Composition immunostimulante
AU740133B2 (en) * 1997-05-19 2001-11-01 Koken Co., Ltd. Immunopotentiating composition
WO1999009956A1 (fr) * 1997-08-29 1999-03-04 Corixa Corporation Agents bioactifs encapsules a liberation rapide servant a provoquer ou a potentialiser une reponse immunitaire, et procedes d'utilisation de ceux-ci
EP1579851A3 (fr) * 1997-08-29 2009-09-02 Corixa Corporation Agents bioactifs encapsulés à libération rapide servant à initier ou à potentialiser une réponse immunitaire, et procédés d'utilisation de ceux-ci
WO1999047588A1 (fr) * 1998-03-14 1999-09-23 Cenes Drug Delivery Limited Production de microparticules
US7285539B2 (en) 1999-10-27 2007-10-23 Chiron Corporation Activation of HCV-specific T cells
US7501134B2 (en) * 2002-02-20 2009-03-10 Novartis Vaccines And Diagnostics, Inc. Microparticles with adsorbed polypeptide-containing molecules
WO2003080033A1 (fr) * 2002-03-21 2003-10-02 Jagotec Ag Microparticules
US6936278B2 (en) 2002-03-21 2005-08-30 Jagotec Ag Microparticles

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