USRE40786E1 - Vaccines against intracellular pathogens using antigens encapsulated within biodegradable-biocompatible microspheres - Google Patents

Vaccines against intracellular pathogens using antigens encapsulated within biodegradable-biocompatible microspheres Download PDF

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USRE40786E1
USRE40786E1 US09/586,747 US58674700A USRE40786E US RE40786 E1 USRE40786 E1 US RE40786E1 US 58674700 A US58674700 A US 58674700A US RE40786 E USRE40786 E US RE40786E
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vaccine
microspheres
composition described
immunostimulating
hiv
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Paul R. Burnett
John E. van Hamont
Robert H. Reid
Jean A. Setterstrom
Thomas C. Van Cott
Deborah L. Birx
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United States Department of the Army
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Priority claimed from US08/242,960 external-priority patent/US5693343A/en
Priority claimed from US08/598,874 external-priority patent/US5762965A/en
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    • 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
    • A61K9/48—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)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
    • C07K14/245—Escherichia (G)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

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  • This invention relates to parenteral and mucosal vaccines against diseases cause by intracellular pathogens using antigens encapsulated within biodegradable-biocompatible microspheres(matrix).
  • HIV-1 infection there is insufficient information at this time regrading the virus and its interactions with the human immune system to permit the rational design of a preventive vaccine.
  • candidate HIV vaccines tested to date fail to elicit antibodies capable of neutralizing wild-type HIV-1 or binding to native HIV-1 proteins, fail to induce a substantial population of effector cells capable of destroying HIV-1-infected cells, and fail to induce significant responses at mucosal surfaces.
  • a possible approach to overcoming these problems is to identify a native protein, accessible to the immune system on the surface of both free virus and infected cells, and present it to the immune system (systemic and mucosal) encapsulated in microspheres to protect and augment its immunogenicity.
  • FIG. 1 indicates controlcytotoxic T lymphocyte induction in mice immunized with rgp 160.
  • This invention relates to a novel pharmaceutical composition, a microcapsule/sphere formulation, which comprises an antigen encapsulated within a biodegradable polymeric matrix, such as poly(DL-lactide co glycolide) (PLG), wherein the molecular weight of the PLG is about 4 , 000 to 100 , 000 daltons and wherein the relative ratio between the lactide and glycolide component of the PLG is within the range of 52:48 to 0:100, and its use, as a vaccine, in the effective induction of antiviral immune responses comprising both virus-specific cytotoxic T lymphocytes and antibodies reactive against native viral antigens.
  • a biodegradable polymeric matrix such as poly(DL-lactide co glycolide) (PLG)
  • PLG poly(DL-lactide co glycolide)
  • Non-CD4-binding, baculo-expressed, recombinant gp 160 IIIB was obtained from MicroGeneSys (Meriden, Conn.).
  • CD4-binding, oligomeric gp 160 CDC451 was obtained from Advanced BioScience Laboratories (Kensington, Md.).
  • Microencapsulation of immunogens PLG microspheres ranging from 1 nanometer to 20 um in diameter and containing a 0.5 to 1.0% antigen core load were prepared by a solvent extractive method. 0 . 5 to 5 . 0 % by weight antigen core load could also be used.
  • the solvent extraction method involves dissolving the viral antigen and sucrose (1:4 ratio w:w) in 1 ml of deionized water. This solution is flash frozen and lyophilized. The resulting antigen-loaded sucrose particles are resuspended in acetonitrile and mixed into PLG copolymer dissolved in acetonitrile.
  • This antigen-polymer mixture is then emulsifyed into heavy mineral oil, transferred into heptane and mixed for 30 min to extract the oil and acetonitrile from the nascent spheres.
  • the spheres are harvested by centrifugation, washed three times in heptane and dried overnight under vacuum. Microsphere size was determined by both light and scanning electron microscopy.
  • the antigen core load was determined by quantitative amino acid analysis of the microspheres following complete hydrolysis in 6N hydrochloric acid.
  • PLG microspheres loaded with native (oligomeric) gp 160 were suspended in phosphate-buffered saline, pH 7.4 (PBS), incubated at 37 C. for 3 h, and then at 4 C overnight. The microspheres were then sedimented by centrifugation (2 min at 200 ⁇ g), the supernatants harvested, and the released gp 160 assayed for binding to CD4 and recognition by HIV-positive patient serum by surface plasmon resonance (described below). A sample of the stock protein used for microencapsulation was assayed for comparison.
  • HIV-seronegative, 8-10 week old NZW rabbits were immunized intramuscularly with rgp 160- or o-gp 160-loaded PLG microspheres suspended in PBS or with alum-adjuvanted rgp 160 in PBS.
  • mice were immunized subcutaneously with rgp 160-loaded PLG microspheres suspend in PBS or with alum-adjuvanted rgp 160 in PBS.
  • the mice in all groups received 10 ug of immunogen on days 0 and 21.
  • FIG. 1 shows the data from a cytotoxic T lymphocyte (CTL) assay performed on the speen cells of mice which had had been previously immunized with either HIV-1 envelope protein encapsulated in PLG microspheres (dark squares) or the same protein administered in a conventional way with alum adjuvant (dark diamonds).
  • CTL cytotoxic T lymphocyte
  • FIG. 2 shows the results of an assay designed to measure the relative binding of antibodies to native vs denatured viral protein.
  • Microencapsulation of immunogens PLG microspheres ranging from 1 to 15 um in diameter and containing a 0.5 to 1.0% antigen core load were prepared by a solvent evaporation method.
  • the solvent evaporation method involves emulsifying the viral antigen dissolved in deionized water into poly(DL-lactide-co-glycolide) polymer dissolved in methylene chloride. This emulsion is mixed into 0.9% polyvinyl alcohol and stirred. After 10 min of stirring, 0.35 l of water is added and gentle mixing is continued for 1.5 h. The resulting spheres are harvested by centrifugation, washed three times in distilled water, and dried overnight under vacuum. Microsphere size was determined by both light and scanning electron microscopy. The antigen core load was determined by quantitative amino acid analysis of the microspheres following complete hydrolysis in 6N hydrochloric acid.

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Abstract

This invention relates to parenteral and mucosal vaccines against diseases caused by intercellular pathogens using antigens encapsulated within a biodegradable-biocompatible microspheres(matrix).

Description

II. CROSS REFERENCE
This application is a continuation-in-part of U.S. patent application Ser. No. 08/242,960, filed May 16, 1994, pending; which in turn is a continuation-in-part of U.S. patent application Ser. No. 07/867,301 filed Apr. 10, 1992, now U.S. Pat. No. 5,417,986 which in turn is a continuation-in-part of U.S. patent application Ser. No. 07/805,721 filed Nov. 21, 1991; now abandoned, which in turn is a continuation-in-part of U.S. patent application Ser. No. 07/690,485 filed Apr. 24, 1991, now abandoned; which in turn is a continuation-in-part of U.S. patent application Ser. No. 07/521,945 filed May 11, 1990, now abandoned. Additionally, this application is a continuation-in-part of U.S. patent application Ser. No. 08/446,149 filed May 22, 1995, pending; which in turn is a continuation of U.S. patent application Ser. No. 06,590,308 filed Mar. 16, 1984, now abandoned.
I. GOVERNMENT INTEREST
The invention descried herein may be manufactured, licensed and used by or for governmental purposes without the payment of any royalties to us thereon.
III. FIELD OF THE INVENTION
This invention relates to parenteral and mucosal vaccines against diseases cause by intracellular pathogens using antigens encapsulated within biodegradable-biocompatible microspheres(matrix).
IV. BACKGROUND OF THE INVENTION
Most infections by viruses and other intracellular pathogens are countered in the human host by a combination of humoral (antibody) and cellular (lymphocyte and phagocyte) immune effectors. Although the precise identity of immune effectors capable of protecting the host against some chronic intracellular pathogens (e.g. HIV-1) remains unknown, attempts to develop preventive and therapeutic vaccines still focus on the induction of appropriate humoral and cellular immune responses. Furthermore, since most human viral pathogens (including HIV-1) are transmitted across mucosal surfaces, it is important that vaccines induce such responses locally (at the mucosal surface) as well as systemically and that they are durable for long-term protection.
The issues of durability and mucosal immunogenicity have been previously addressed by encapsulating vaccine antigens in appropriately-sized biodegradable, biocompatible microspheres made of lactide/glycolide copolymer (the same materials used in resorbable sutures). It has been shown that such microspheres can be made to release their load in a controlled manner over a prolonged period of time and can facilitate the interaction of their contents with the local immune system when administrated mucosally.
In the case of HIV-1 infection, there is insufficient information at this time regrading the virus and its interactions with the human immune system to permit the rational design of a preventive vaccine. However, it has been noted that many candidate HIV vaccines tested to date fail to elicit antibodies capable of neutralizing wild-type HIV-1 or binding to native HIV-1 proteins, fail to induce a substantial population of effector cells capable of destroying HIV-1-infected cells, and fail to induce significant responses at mucosal surfaces. A possible approach to overcoming these problems (applicable to both HIV-1 and other chronic intracellular pathogens) is to identify a native protein, accessible to the immune system on the surface of both free virus and infected cells, and present it to the immune system (systemic and mucosal) encapsulated in microspheres to protect and augment its immunogenicity.
V. DESCRIPTION OF DRAWINGS
FIG. 1 indicates controlcytotoxic T lymphocyte induction in mice immunized with rgp 160; and
FIG. 2 indicates “Native”/denatured rgp 120 (IIIB) Binding Ratios.
 V. DESCRIPTION OF THE INVENTION
This invention relates to a novel pharmaceutical composition, a microcapsule/sphere formulation, which comprises an antigen encapsulated within a biodegradable polymeric matrix, such as poly(DL-lactide co glycolide) (PLG), wherein the molecular weight of the PLG is about 4,000 to 100,000 daltons and wherein the relative ratio between the lactide and glycolide component of the PLG is within the range of 52:48 to 0:100, and its use, as a vaccine, in the effective induction of antiviral immune responses comprising both virus-specific cytotoxic T lymphocytes and antibodies reactive against native viral antigens. In the practice of this invention, applicants found that when a complex (oligomeric) native envelope protein of HIV-1 was encapsulated in PLG microspheres, it retained its native antigenicity and function upon its release in vitro. Furthermore, when used as a vaccine in animals, this product elicited HIV-specific cytotoxic T lymphocytes and antibodies reactive with native (oligomeric) HIV-1 envelope protein.
The following examples illustrate the invention:
EXAMPLE 1
Materials and Methods
Immunogens, Non-CD4-binding, baculo-expressed, recombinant gp 160IIIB (rgp 160) was obtained from MicroGeneSys (Meriden, Conn.). CD4-binding, oligomeric gp 160 CDC451 (o-gp 160) was obtained from Advanced BioScience Laboratories (Kensington, Md.).
Microencapsulation of immunogens: PLG microspheres ranging from 1 nanometer to 20 um in diameter and containing a 0.5 to 1.0% antigen core load were prepared by a solvent extractive method. 0.5 to 5.0% by weight antigen core load could also be used. The solvent extraction method involves dissolving the viral antigen and sucrose (1:4 ratio w:w) in 1 ml of deionized water. This solution is flash frozen and lyophilized. The resulting antigen-loaded sucrose particles are resuspended in acetonitrile and mixed into PLG copolymer dissolved in acetonitrile. This antigen-polymer mixture is then emulsifyed into heavy mineral oil, transferred into heptane and mixed for 30 min to extract the oil and acetonitrile from the nascent spheres. The spheres are harvested by centrifugation, washed three times in heptane and dried overnight under vacuum. Microsphere size was determined by both light and scanning electron microscopy. The antigen core load was determined by quantitative amino acid analysis of the microspheres following complete hydrolysis in 6N hydrochloric acid.
Analysis of immunogen spontaneously released from microspheres in vitro by binding to soluble CD4 and recognition by HIV-positive patient serum. PLG microspheres loaded with native (oligomeric) gp 160 were suspended in phosphate-buffered saline, pH 7.4 (PBS), incubated at 37 C. for 3 h, and then at 4 C overnight. The microspheres were then sedimented by centrifugation (2 min at 200×g), the supernatants harvested, and the released gp 160 assayed for binding to CD4 and recognition by HIV-positive patient serum by surface plasmon resonance (described below). A sample of the stock protein used for microencapsulation was assayed for comparison.
Immunization of animals. HIV-seronegative, 8-10 week old NZW rabbits were immunized intramuscularly with rgp 160- or o-gp 160-loaded PLG microspheres suspended in PBS or with alum-adjuvanted rgp 160 in PBS. Groups receiving rgp 160-loaded microspheres (n=2) were primed with 50 ug of immunogen on day 0 and boosted with 25 ug on day 42. Groups receiving o-gp 160-loaded microspheres (n=3) were primed with 70 ug of immunogen on day 0 and boosted with 35 ug on day 56. Groups receiving alum-adjuvanted rgp 160 (n=2) got 85 ug of immunogen on days 0, 7, and 28.
BALB/c mice were immunized subcutaneously with rgp 160-loaded PLG microspheres suspend in PBS or with alum-adjuvanted rgp 160 in PBS. The mice in all groups (n=4) received 10 ug of immunogen on days 0 and 21.
Determination of the ratio of antibody binding to “native”/denatured rgp 120IIIB measured by surface plasmon resonance (SPR). Real-time binding interactions between ligand (gp 120 covalently linked to a biosensor matrix) and ligate (Abs in solution) were measured using surface plasmon resonance (BIAcore, Pharmacia Biosensor, Piscataway, N.J.). “Native”rgp 120(IIIB) (Genentech, South San Francisco, Calif.) or reduced, carboxymethylated (denatured) rgp 120(IIIB) (Genentech) was covalently linked to the biosensor dextran matrix. Sera and mAbs were diluted in HBS running buffer (10 mM HEPES, 150 mM NaCl, 3.4 mM EDTA, 0.05% (BIAcore) surfactant P20, pH 7.4) and injected through the dextran matrices at a flow rate of 5 ul/min. The binding value of each serum or mAb was measured in resonance units (RU), and the “native”/denatured gp 120 ratios were determined by dividing the corresponding RU values and correcting for small differences in matrix concentration. Control included an HIV-positive patient serum and mAb 1c1.
Assessment of HIV-specific cell-mediated immunity in immunized mice by secondary CTL assay. The spleens of BALB/c mice immunized on days 0 and 21 were harvested and single cell suspensions prepared aseptically in complete RPMI medium on day 35. The cells were then pooled within experimental groups (n=4), underlay with ficoll, centrifuged 30 min at 450×g (RT), washed, and resuspended in complete RPMI medium. Following a 1 h stimulation with peptide p18 (1 uM) at 37° C., the cell suspensions were diluted with complete RPMI supplemented with 2ME (1:1000) and transferred to flasks for a 6 day incubation at 37° C. After 2 days, recombinant IL-2 (10 u/ml) was added to all flasks. On day 6, P815 target cells were pulsed with peptide p18 (1 uM) or with nothing (control) in PBS supplemented with 0.1% BSA. 3×10A6 target cells were labelled with 300 uCi of 51Cr, washed, and plated out with the effector cells at effector:traget (E:T) ratios of 45:1, 15:1, 5:1, and 1.7:1. After a 6 h incubation at 37° C., the supernatants were harvested and counted, and % specific lysis was calculated.
Results
Comparison of the native (oligomeric) gp 160 prior to microencapsulation and following spontaneous release from PLG microspheres showed the two to be essentially indistinguishable in terms of their binding to CD4 and recognition by HIV-positive patient serum. (Table 1). This retention of conformation-dependent binding shows that structure of the antigen is not appreciably altered by the microencapsulation process.
FIG. 1 shows the data from a cytotoxic T lymphocyte (CTL) assay performed on the speen cells of mice which had had been previously immunized with either HIV-1 envelope protein encapsulated in PLG microspheres (dark squares) or the same protein administered in a conventional way with alum adjuvant (dark diamonds). These data indicate that microencapsulation of HIV-1 envelope protein in PLG microspheres results in a vaccine that induces significantly greater anti-HIV CTL activity than does alum-adjuvanted vaccine. The open symbol groups represent controls run to assure that the activity being measured is virus-specific.
FIG. 2 shows the results of an assay designed to measure the relative binding of antibodies to native vs denatured viral protein. These data show that rabbits immunized with a non-native HIV-1 protein encapsulated in PLG (#5 and 6) develop antibodies which show greater binding to denatured (vs native) protein (indicated by a ratio<1). On the other hand, rabbits immunized with a native HIV-1 protein encapsulated in PLG microspheres (#10-12) develop antibodies which show greater binding to native viral protein (indicated by ratio>1). This retention of each proteins antigenicity constitutes an additional piece of evidence that the structure of antigens loaded in PLG microspheres are preserved.
EXAMPLE 2
Materials and Methods
This experiment was similar to that described in Example 1 except for the method of microencapsulation employed.
Microencapsulation of immunogens: PLG microspheres ranging from 1 to 15 um in diameter and containing a 0.5 to 1.0% antigen core load were prepared by a solvent evaporation method. The solvent evaporation method involves emulsifying the viral antigen dissolved in deionized water into poly(DL-lactide-co-glycolide) polymer dissolved in methylene chloride. This emulsion is mixed into 0.9% polyvinyl alcohol and stirred. After 10 min of stirring, 0.35 l of water is added and gentle mixing is continued for 1.5 h. The resulting spheres are harvested by centrifugation, washed three times in distilled water, and dried overnight under vacuum. Microsphere size was determined by both light and scanning electron microscopy. The antigen core load was determined by quantitative amino acid analysis of the microspheres following complete hydrolysis in 6N hydrochloric acid.
Results
Analysis of spontaneously released antigen showed it to retain its CD4 binding capacity. Its native antigenicity (recognition by the serum of an HIV-positive patient) was only slightly less than that of the antigen prior to encapsulation and following spontaneous release from microspheres produced by a solvent extraction method (Table 1).
The results of immunizing animals with either non-native (denatured) or native oligomeric gp 160 in PLG microspheres produced by a solvent evaporation method were essentially indistinguishable from those obtained using microspheres produced by a solvent extraction method (example 1). Microencapsulated antigen induced significantly greater CTL activity than antigen administered in a conventional alum-adjuvanted formulation. Furthermore, preservation of the structure of PLG-microencapsulated antigens is supported by the findings of preferential binding of antibodies elicited by microspheres loaded with denatured antigen to denatured gp 120 (FIGS. 2, 3, and 4) (FIG. 2, nos. 3 and 4) and the preferred binding of antibodies elicited by microspheres loaded with native (oligomeric) antigen to native gp 120 (FIGS. 2, 7-8) (FIG. 2, nos. 7 and 8).
TABLE 1
BIA (released o-gp160)
Capture o-gp160-451 (stock vs microsphere-released)
on tvc 391 fc3/fc4 sCD4 (4 mg/m)
1 ul/min flow rate foe o-gp160 ini.: 5 ul/min for all others
ligate RU HIV+/sCD4 (RU ratio)
gp120-MN 1:10 3286
HIV+ 1:100 54
NHS 1:100 3
HIV+ pool 1:100 47
o-gp160 (tvc281) 1772
HIV+ 3259 1.84
tvc281 1848
NHS −36
tvc281 1762
HIV+ pool 2597 1.47
tvc281-PLG-EV 3342
HIV+ 4594 1.37
tvc281 3222
NHS 7
tvc281 3210
HIV+ pool 3336 1.04
tvc281-PLG-EX 1855
HIV+ 3760 2.04
tvc281 1839
NHS 2
tvc281 1850
HIV+ pool 2745 1.48
gp120-MN 1:10 2914
HIV+ 1:100 14
NHS 1:100 −2
HIV+ pool 1:100 14
tvc281 1099
HIV+ 1083 0.99
tvc281 1022
HIV+ pool 1395 1.36
tvc281-PLG-EV 1595
HIV+ 1322 0.83
tvc281 1535
HIV+ pool 1781 1.16
In view of the above it will be seen that the objects of the invention are achieved. As various changes could be made in the above materials and methods without departing from the scope of the invention, it is intended that all matter contained in the above description shall be interpreted as illustrative and not limiting.

Claims (14)

1. An immunostimulating composition comprising encapsulating encapsulated microspheres comprised of (a) a biodegradable-biocompatible poly(DL-lactide-co-glycolideas poly(DL-lactide-co-glycolide) as the bulk matrix produced by a solvent evaporation process wherein the molecular weight of the copolymer is between 4,000 to 100,000 daltons and (b) an immunogenic substance consisting of a conformationally native subunit of chronic intracellular pathogen which, in the course of natural infection with that pathogen, is exposed to the host immune system on the surface of free pathogen and/or pathogen-infected cells.
2. The immunostimulating composition described in claim 1 wherein the immunogenic substance is an antigen and the antigen is pre-encapsulated into a conformationally stabilizing hydrophilic matrix consisting of an appropriate mono, di- or tri-saccharide or other carbohydrate susbstance substance by lyophilization prior to its final encapsulation into the PLG microsphere by a solvent extraction process employing acetonitrile as the polymer solvent, mineral oil as the emulsion's external phase, and heptane as the extractant.
3. The immunostimulating compositions composition described in claims claim 1 or 2 wherein the immunogenic substance is a native (oligomeric)HIV-1 envelope antigen that is conformationally stabilized by the polymer matrix and serves to elicit in animals the production of HIV specific cytotoxic T lumphocytes lymphocytes and antibodies preferentially reactive against native HIV-1 envelope antigen.
4. The immunostimulating compositions composition described in claim 3 wherein the amount of said immunogenic substance within the microcapsule comprises between 0.5% to 5.0% of the weight of said composition.
5. The immunostimulating compositions describe composition described in claim 4 wherein the relative ratio between the amount of the lactide:glycolide components of said matrix is within the range of 52:48 to 0:100.
6. The immunostimulating compositions composition described in claim 5 wherein the molecular weight of said copolymer is between 4,000 to 50,000 daltons.
7. A vaccine consisting of a blend of the immunostimulating compositions described in claims 5 or 6 composition of claim 5.
8. The immunostimulating compositions composition described in claim 5, employed as a parentally parenterally administered vaccine wherein the diameter size range of said vaccine microspheres lies between 1 nanometer and 20 microns.
9. The immunostimulating compositions composition described in claim 5, employed as a mucosal vaccine wherein the size of more than 50% (by volume) of said vaccine microspheres is between 5 microns to 10 microns in diameter.
10. A composition in accordance with claim 1 wherein the microspheres further contain a pharmaceutically-acceptable adjuvant.
11. A vaccine consisting of a blend of the immunostimulating compositions described in claims 5 or composition of claim 6.
12. The immunostimulating compositions composition described in claim 6 employed as a parentally parenterally administered vaccine wherein the diameter size range of said vaccine microspheres lies between 1 nanometer and 20 microns.
13. The immunostimulating compositions composition described in claim 7 employed as a parentally parenterally administered vaccine wherein the diameter size range of said vaccine microspheres lies between nanogram nanometer and 20 microns.
14. The immunostimulating compositions composition described in claim 6 employed as a mucosal vaccine wherein the size of more than 50% (by volume) of said vaccine microspheres is between 5 microns to 10 microns in diameter.
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US59030884A 1984-03-16 1984-03-16
US52194590A 1990-05-11 1990-05-11
US69048591A 1991-04-24 1991-04-24
US80572191A 1991-11-21 1991-11-21
US07/867,301 US5417986A (en) 1984-03-16 1992-04-10 Vaccines against diseases caused by enteropathogenic organisms using antigens encapsulated within biodegradable-biocompatible microspheres
US08/242,960 US5693343A (en) 1984-03-16 1994-05-16 Microparticle carriers of maximal uptake capacity by both M cells and non-M cells
US44614995A 1995-05-22 1995-05-22
US08/598,874 US5762965A (en) 1984-03-16 1996-02-09 Vaccines against intracellular pathogens using antigens encapsulated within biodegradble-biocompatible microspheres
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Citations (45)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3540444A (en) 1968-01-15 1970-11-17 Scherer Corp R P Plastic ampoule for use with hypodermic injector
US3773919A (en) 1969-10-23 1973-11-20 Du Pont Polylactide-drug mixtures
US3788315A (en) 1971-04-20 1974-01-29 S Laurens Disposable cutaneous transjector
US4166800A (en) 1977-08-25 1979-09-04 Sandoz, Inc. Processes for preparation of microspheres
US4384975A (en) 1980-06-13 1983-05-24 Sandoz, Inc. Process for preparation of microspheres
US4389330A (en) * 1980-10-06 1983-06-21 Stolle Research And Development Corporation Microencapsulation process
US4530840A (en) 1982-07-29 1985-07-23 The Stolle Research And Development Corporation Injectable, long-acting microparticle formulation for the delivery of anti-inflammatory agents
US4542025A (en) 1982-07-29 1985-09-17 The Stolle Research And Development Corporation Injectable, long-acting microparticle formulation for the delivery of anti-inflammatory agents
US4585482A (en) 1984-05-25 1986-04-29 Southern Research Institute Long-acting biocidal compositions and method therefor
US4622244A (en) 1979-09-04 1986-11-11 The Washington University Process for preparation of microcapsules
US4637905A (en) 1982-03-04 1987-01-20 Batelle Development Corporation Process of preparing microcapsules of lactides or lactide copolymers with glycolides and/or ε-caprolactones
US4675189A (en) 1980-11-18 1987-06-23 Syntex (U.S.A.) Inc. Microencapsulation of water soluble active polypeptides
US4798786A (en) 1982-05-06 1989-01-17 Stolle Research And Development Corporation Living cells encapsulated in crosslinked protein
US4835139A (en) 1983-09-23 1989-05-30 Debiopharm S.A. Process for increasing the antagonistic effect of peptidic compounds on hormone-dependent diseases
US4863735A (en) 1985-02-19 1989-09-05 Massachusetts Institute Of Technology Biodegradable polymeric drug delivery system with adjuvant activity
US4897268A (en) 1987-08-03 1990-01-30 Southern Research Institute Drug delivery system and method of making the same
US4938763A (en) 1988-10-03 1990-07-03 Dunn Richard L Biodegradable in-situ forming implants and methods of producing the same
US4941880A (en) 1987-06-19 1990-07-17 Bioject, Inc. Pre-filled ampule and non-invasive hypodermic injection device assembly
US5000886A (en) 1987-05-26 1991-03-19 American Cyanamid Company Silicone-hardened pharmaceutical microcapsules and process of making the same
US5019096A (en) 1988-02-11 1991-05-28 Trustees Of Columbia University In The City Of New York Infection-resistant compositions, medical devices and surfaces and methods for preparing and using same
US5059187A (en) 1988-11-30 1991-10-22 Dey Laboratories, Inc. Method for the cleansing of wounds using an aerosol container having liquid wound cleansing solution
US5064413A (en) 1989-11-09 1991-11-12 Bioject, Inc. Needleless hypodermic injection device
US5075109A (en) 1986-10-24 1991-12-24 Southern Research Institute Method of potentiating an immune response
US5102872A (en) 1985-09-20 1992-04-07 Cetus Corporation Controlled-release formulations of interleukin-2
US5129825A (en) 1987-12-21 1992-07-14 Discko John Jr Dental syringe and capsule for use therewith
US5133701A (en) 1989-04-06 1992-07-28 Sang In Han Disposable pressure wound irrigation device
WO1992022654A1 (en) * 1991-06-11 1992-12-23 Microgenesys, Inc. Vaccine and treatment method of human immunodeficiency virus infection
US5236355A (en) 1988-12-22 1993-08-17 American Cyanamid Company Apparatus for the treatment of periodontal disease
US5290494A (en) 1990-03-05 1994-03-01 Board Of Regents, The University Of Texas System Process of making a resorbable implantation device
EP0052510B2 (en) 1980-11-18 1994-10-19 Syntex (U.S.A.) Inc. Microencapsulation of water soluble polypeptides
US5360610A (en) 1990-05-16 1994-11-01 Southern Research Institute Method for stimulating nerve fiber growth
US5384133A (en) 1986-08-11 1995-01-24 Innovata Biomed Limited Pharmaceutical formulations comprising microcapsules
US5407609A (en) 1989-05-04 1995-04-18 Southern Research Institute Microencapsulation process and products therefrom
WO1995011010A1 (en) * 1993-10-22 1995-04-27 Genentech, Inc. Methods and compositions for microencapsulation of antigens for use as vaccines
US5417986A (en) 1984-03-16 1995-05-23 The United States Of America As Represented By The Secretary Of The Army Vaccines against diseases caused by enteropathogenic organisms using antigens encapsulated within biodegradable-biocompatible microspheres
US5429822A (en) 1992-03-13 1995-07-04 Cambridge Scientific, Inc. Biodegradable bursting release system
US5500228A (en) 1987-05-26 1996-03-19 American Cyanamid Company Phase separation-microencapsulated pharmaceuticals compositions useful for alleviating dental disease
US5538739A (en) 1989-07-07 1996-07-23 Sandoz Ltd. Sustained release formulations of water soluble peptides
US5639480A (en) 1989-07-07 1997-06-17 Sandoz Ltd. Sustained release formulations of water soluble peptides
US5643605A (en) 1993-10-25 1997-07-01 Genentech, Inc. Methods and compositions for microencapsulation of adjuvants
US5648096A (en) 1992-10-26 1997-07-15 Schwarz Pharma Ag Process for the production of microcapsules
US5650173A (en) 1993-11-19 1997-07-22 Alkermes Controlled Therapeutics Inc. Ii Preparation of biodegradable microparticles containing a biologically active agent
US5693343A (en) 1984-03-16 1997-12-02 The United States Of America As Represented By The Secretary Of The Army Microparticle carriers of maximal uptake capacity by both M cells and non-M cells
US5762965A (en) 1984-03-16 1998-06-09 The United States Of America As Represented By The Secretary Of The Army Vaccines against intracellular pathogens using antigens encapsulated within biodegradble-biocompatible microspheres
US5811128A (en) 1986-10-24 1998-09-22 Southern Research Institute Method for oral or rectal delivery of microencapsulated vaccines and compositions therefor

Patent Citations (51)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3540444A (en) 1968-01-15 1970-11-17 Scherer Corp R P Plastic ampoule for use with hypodermic injector
US3773919A (en) 1969-10-23 1973-11-20 Du Pont Polylactide-drug mixtures
US3788315A (en) 1971-04-20 1974-01-29 S Laurens Disposable cutaneous transjector
US4166800A (en) 1977-08-25 1979-09-04 Sandoz, Inc. Processes for preparation of microspheres
US4622244A (en) 1979-09-04 1986-11-11 The Washington University Process for preparation of microcapsules
US4384975A (en) 1980-06-13 1983-05-24 Sandoz, Inc. Process for preparation of microspheres
US4389330A (en) * 1980-10-06 1983-06-21 Stolle Research And Development Corporation Microencapsulation process
EP0052510B2 (en) 1980-11-18 1994-10-19 Syntex (U.S.A.) Inc. Microencapsulation of water soluble polypeptides
US4675189A (en) 1980-11-18 1987-06-23 Syntex (U.S.A.) Inc. Microencapsulation of water soluble active polypeptides
US4637905A (en) 1982-03-04 1987-01-20 Batelle Development Corporation Process of preparing microcapsules of lactides or lactide copolymers with glycolides and/or ε-caprolactones
US4798786A (en) 1982-05-06 1989-01-17 Stolle Research And Development Corporation Living cells encapsulated in crosslinked protein
US4542025A (en) 1982-07-29 1985-09-17 The Stolle Research And Development Corporation Injectable, long-acting microparticle formulation for the delivery of anti-inflammatory agents
US4530840A (en) 1982-07-29 1985-07-23 The Stolle Research And Development Corporation Injectable, long-acting microparticle formulation for the delivery of anti-inflammatory agents
US4835139A (en) 1983-09-23 1989-05-30 Debiopharm S.A. Process for increasing the antagonistic effect of peptidic compounds on hormone-dependent diseases
US5417986A (en) 1984-03-16 1995-05-23 The United States Of America As Represented By The Secretary Of The Army Vaccines against diseases caused by enteropathogenic organisms using antigens encapsulated within biodegradable-biocompatible microspheres
US5693343A (en) 1984-03-16 1997-12-02 The United States Of America As Represented By The Secretary Of The Army Microparticle carriers of maximal uptake capacity by both M cells and non-M cells
US5762965A (en) 1984-03-16 1998-06-09 The United States Of America As Represented By The Secretary Of The Army Vaccines against intracellular pathogens using antigens encapsulated within biodegradble-biocompatible microspheres
US4585482A (en) 1984-05-25 1986-04-29 Southern Research Institute Long-acting biocidal compositions and method therefor
US4863735A (en) 1985-02-19 1989-09-05 Massachusetts Institute Of Technology Biodegradable polymeric drug delivery system with adjuvant activity
US5102872A (en) 1985-09-20 1992-04-07 Cetus Corporation Controlled-release formulations of interleukin-2
US5384133A (en) 1986-08-11 1995-01-24 Innovata Biomed Limited Pharmaceutical formulations comprising microcapsules
US5820883A (en) 1986-10-24 1998-10-13 Southern Research Institute Method for delivering bioactive agents into and through the mucosally-associated lymphoid tissues and controlling their release
US5814344A (en) 1986-10-24 1998-09-29 Southern Research Institute Method for delivering bioactive agents into and through the mucosally associated lymphoid tissues and controlling their release
US5811128A (en) 1986-10-24 1998-09-22 Southern Research Institute Method for oral or rectal delivery of microencapsulated vaccines and compositions therefor
US5075109A (en) 1986-10-24 1991-12-24 Southern Research Institute Method of potentiating an immune response
US5853763A (en) 1986-10-24 1998-12-29 Southern Research Institute Method for delivering bioactive agents into and through the mucosally-associated lymphoid tissue and controlling their release
US5000886A (en) 1987-05-26 1991-03-19 American Cyanamid Company Silicone-hardened pharmaceutical microcapsules and process of making the same
US5500228A (en) 1987-05-26 1996-03-19 American Cyanamid Company Phase separation-microencapsulated pharmaceuticals compositions useful for alleviating dental disease
US4941880A (en) 1987-06-19 1990-07-17 Bioject, Inc. Pre-filled ampule and non-invasive hypodermic injection device assembly
US4897268A (en) 1987-08-03 1990-01-30 Southern Research Institute Drug delivery system and method of making the same
US5129825A (en) 1987-12-21 1992-07-14 Discko John Jr Dental syringe and capsule for use therewith
US5019096A (en) 1988-02-11 1991-05-28 Trustees Of Columbia University In The City Of New York Infection-resistant compositions, medical devices and surfaces and methods for preparing and using same
US4938763B1 (en) 1988-10-03 1995-07-04 Atrix Lab Inc Biodegradable in-situ forming implants and method of producing the same
US4938763A (en) 1988-10-03 1990-07-03 Dunn Richard L Biodegradable in-situ forming implants and methods of producing the same
US5278202A (en) 1988-10-03 1994-01-11 Atrix Laboratories, Inc. Biodegradable in-situ forming implants and methods of producing the same
US5059187A (en) 1988-11-30 1991-10-22 Dey Laboratories, Inc. Method for the cleansing of wounds using an aerosol container having liquid wound cleansing solution
US5236355A (en) 1988-12-22 1993-08-17 American Cyanamid Company Apparatus for the treatment of periodontal disease
US5133701A (en) 1989-04-06 1992-07-28 Sang In Han Disposable pressure wound irrigation device
US5407609A (en) 1989-05-04 1995-04-18 Southern Research Institute Microencapsulation process and products therefrom
US5538739A (en) 1989-07-07 1996-07-23 Sandoz Ltd. Sustained release formulations of water soluble peptides
US5639480A (en) 1989-07-07 1997-06-17 Sandoz Ltd. Sustained release formulations of water soluble peptides
US5688530A (en) 1989-07-07 1997-11-18 Novartis Ag Sustained release formulations of water soluble peptides
US5064413A (en) 1989-11-09 1991-11-12 Bioject, Inc. Needleless hypodermic injection device
US5290494A (en) 1990-03-05 1994-03-01 Board Of Regents, The University Of Texas System Process of making a resorbable implantation device
US5360610A (en) 1990-05-16 1994-11-01 Southern Research Institute Method for stimulating nerve fiber growth
WO1992022654A1 (en) * 1991-06-11 1992-12-23 Microgenesys, Inc. Vaccine and treatment method of human immunodeficiency virus infection
US5429822A (en) 1992-03-13 1995-07-04 Cambridge Scientific, Inc. Biodegradable bursting release system
US5648096A (en) 1992-10-26 1997-07-15 Schwarz Pharma Ag Process for the production of microcapsules
WO1995011010A1 (en) * 1993-10-22 1995-04-27 Genentech, Inc. Methods and compositions for microencapsulation of antigens for use as vaccines
US5643605A (en) 1993-10-25 1997-07-01 Genentech, Inc. Methods and compositions for microencapsulation of adjuvants
US5650173A (en) 1993-11-19 1997-07-22 Alkermes Controlled Therapeutics Inc. Ii Preparation of biodegradable microparticles containing a biologically active agent

Non-Patent Citations (21)

* Cited by examiner, † Cited by third party
Title
Biotechnology News, Aug. 22, 1997, vol. 17, No. 20, Topical DNA vaccine elicits immune response.
Brown, A hypothetical model of the foreign antigen blinding site of Class II histocompatibility molecules, Nature, vol. 332, Apr. 28, 1988, pp. 845-850.
Brown, Wonder Drugs' Losing Healing Aura, The Washing Post, Jun. 26, 1995, A section.
Cassels, et al., Analysis of Escherichia coli Colonization Factor Antigen I Linear B-Cell Epitopes, as Determined by Primate Responses, following Protein Sequence Verification, Infection and Immunity, Jun. 1992, pp. 2174-2181, vol. 60, No. 6.
Evans, et al. Purification and Characterization of the CFR/1 Antigen of Enterotoxigenic Escherichia coli, Infection and Immunity, Aug. 1979, pp. 738-748, vol. 25.
Gilding, Biodegradable polymers for use in surgery-polyglycolic/poly (ac c acid) homo-and copolymers: 1, Polymer, vol. 20, Dec. 1979, pp. 1459-1464.
Hall, et al., Purification and Analysis of Colonization Factor Antigen 1, Coli Surface Antigen 1, and Coli Surface ANtigen 3 Fimbriae from Enterotoxigenic Escherichis Coli, Journal of Bacteriology, Nov. 1989, pp. 6372-6374, vol. 171, No. 11.
Jeyanthi, et al., Novel, Burst Free Programmable Biodegradable Microspheres For Controlled Release of Polypeptides, Proceedings Int. Symp. control Release Bioact. Mater. (1996) pp. 351-362/.
Jordi, et al., Analysis of the first two genes of the CSI fimbrial operon in human enterotoxigenic Escherichia coli of serotype 0139: H28, FEMS Microbiology Letters 80, (1991) pp. 265-270.
Karjalainen, et al., Molecular Cloning and Nucleotide Sequence of the Colonization Factor Antigen I Gene of Escherichia coli, Infection and Immunity, Apr. 1989, pp. 1126-1130, vol. 57.
Maister, First Oral AIDS Vaccine Trials Near, BioWorld Today, Tuesday, Apr. 19, 1994, p. 4.
McConnel, et al., Antigenic homology within human enterotoxigenic Esherichia coli fimbrial colonization factor antigens: CFA/I, coli-surface-associated antigens (CS)1, CS2, CS4 and CS17, FEMS Microbiology Letters 61 (1989) 105-108.
Perez-Casal, et al., Gene Encoding the Major Subunit of CS1 Pili of Human Enterotoxigenic Escherichia Coli, Infection and Immunity, Nov., 1990, pp. 3594-3600, vol. 58, No. 11.
Rognan, et al., Molecular Modeling of an Antigenic Complex Between a Viral Peptide and a Class I Major Histocompatibility Glycoprotein, Proteins Structure, Function and Genetics 13 70-85 (1992).
Romagnoli, et al. Peptide-MHC Interaction: A Rational Approach to Vaccine Design, Inter, RE. Immunol. 6, 1990, 00 61-73.
Setterstrom, Controlled Release of Antibiotics From biodegradable Microcapsules For Wound Infection Control, Chemical Abstracts, 1983, pp. 215-226.
Tan, et al., Mapping the Antigenic Epitopes of Human Dihydrofolate Reductase by Systematic Synthesis of Peptides on solid Supports, The Journal of Biological Chemistry, vol. 265, No. 14, Issue of May 15, pp. 8022-8026 (1990).
Van der Zee, Efficient mapping and characterization of a T cell epitope by the simultaneous synthesis of multiple peptides, Eur. J. Immunol. 1989, 19: 43-47.
Wang, et al., Influence of formulation methods on the in vitro controlled release of protein from poly (ester) microspheres Journal of Controlled Release, 17 (1991) 23-32.
Yan, Characterization and morphological analysis of protein-loaded poly(lactide-co-glycolide) microparticles prepared by watewr-in-oil-in-water emulsion technique, Journal of Controlled Release, 32 (1994) 231-241.
Yeh, A novel emulsification-solvent extraction technique for production of protein loaded biodegradable microparticles for vaccine and drug delivery, Journal of Controlled Release 33 (1995) 437-445.

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