WO2006059846A1 - Formulation de proteine mutee sec1, et procede de formulation de celle-ci - Google Patents

Formulation de proteine mutee sec1, et procede de formulation de celle-ci Download PDF

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Publication number
WO2006059846A1
WO2006059846A1 PCT/KR2005/003871 KR2005003871W WO2006059846A1 WO 2006059846 A1 WO2006059846 A1 WO 2006059846A1 KR 2005003871 W KR2005003871 W KR 2005003871W WO 2006059846 A1 WO2006059846 A1 WO 2006059846A1
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formulation
protein
secl
oil
formulation according
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PCT/KR2005/003871
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English (en)
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Myung-Soo Kang
Byoung Sun Chang
Jin Hee Lee
Ki-Young Yoon
Yun-Sik Kim
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Lg Life Sciences, Ltd.
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Publication of WO2006059846A1 publication Critical patent/WO2006059846A1/fr

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    • 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
    • A61K38/164—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • 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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A61K47/38—Cellulose; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • 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
    • A61K9/1617—Organic compounds, e.g. phospholipids, fats
    • 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
    • A61K9/1617—Organic compounds, e.g. phospholipids, fats
    • A61K9/1623—Sugars or sugar alcohols, e.g. lactose; Derivatives thereof; Homeopathic globules
    • 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
    • A61K9/1635—Organic macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
    • 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
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders

Definitions

  • the present invention relates to a formulation of a Staphylococcal enterotoxin Cl
  • the present invention relates to a formulation of an SECl mutant protein comprising an effective amount of an SECl mutant protein, a protein- stabilizing excipient, a carbohydrate-based auxiliary excipient, a lipophilic material, and a biocompatible oil and/or a fatty acid ester-based compound, by which the SECl mutant protein can be easily administered via injection and efficacy and stability thereof are maximized, and a method for formulating the same.
  • Staphylococcal enterotoxin C 1 (SEC 1 ) mutant protein a toxin of Staphylococcus aureus, is a protein in which cysteine, an amino acid at a position 95 of a mutant toxin Cl of Staphylococcus aureus, was substituted with serine, and is known to have a probability of effective application thereof as a vaccine inducing promotion of nonspecific cellular immunity as well as antibody production of specific humoral immunity (Terence N. Turner et al (1992), Infection and Immunity 62(2), pp 694-697; Carolyn J. Hovde et al (1994), Molecular Microbiology 13(5), pp 897-909; and Marcy L. Hoffann et al (1994), Infection and Immunity 62(8), pp 3396-3407).
  • a method for preparing an SECl mutant protein is disclosed in Korean Patent No. 382239, Australian Patent No. 2001-11759 and the like.
  • the SECl mutant protein exhibiting such prevention, symptom alleviation and therapeutic effects of mastitis can be mass- produced using Escherichia coli as a host.
  • the SECl mutant protein also suffers from problems associated with maintenance of protein stability such as protein denaturation upon long-term storage (more than 2 weeks) and aggregation of protein in dispersion media (for example, oil).
  • the SECl mutant protein like ordinary proteins, is also labile to heat, pH, salts and organic solvents (Weiqi Lu et al. PDA L. Pharm. Sci. Tech. 49, 13-19 (1995)).
  • Korean Patent No. 359252 assigned to the present applicant, discloses a method for preparing microparticles of SECl mutant protein using 3% car- boxymethylcellulose and 2% lecithin via spray drying.
  • preparation of the SECl mutant protein by means of spray drying exhibits a low yield of about 10 to 30% and thus is not suitable for commercialization via industrial-scale production.
  • the SECl mutant protein is exposed to a high internal temperature of 50 to 70°C which may cause denaturation of the protein.
  • an SECl mutant protein formulation comprising an effective amount of an SECl mutant protein as an active ingredient, and prepared by mixing the SECl mutant protein with a protein-stabilizing excipient containing particular ingredients, a carbohydrate-based auxiliary excipient and a lipophilic material to prepare solid microparticles and dispersing the resulting microparticles in a biocompatible oil and/or a fatty acid ester- based compound (a dispersion medium), can prevent denaturation occurring upon long-term storage of the SECl mutant protein in a solution state, aggregation in the dispersion media and instability of the protein due to a variety of external factors, and is capable of achieving prevention, symptom alleviation and maximized therapeutic effects of mastitis in the body as well as commercialization thereof via industrial-scale production.
  • the present invention has been completed based on these findings.
  • the present invention has been made in view of the above problems, and it is an object of the present invention to provide a formulation which is capable of easily administering a water soluble SECl mutant protein via injection while maintaining stability thereof and is capable of maintaining activity of the protein for a prolonged period of time in vivo when it is administered.
  • an SECl mutant protein formulation comprising solid microparticles containing 0.001 to 50% by weight of a Staphylococcal enterotoxin Cl (SECl) mutant protein, one of toxins of Staphylococcus aureus, as an active ingredient, 0.1 to 90% by weight of a protein-stabilizing excipient, 0.1 to 90% by weight of a carbohydrate-based auxiliary excipient and 0.1 to 10% by weight of a lipophilic material, dispersed in a biocompatible oil and/or a fatty acid ester-based compound.
  • SECl Staphylococcal enterotoxin Cl
  • the formulation in accordance with the present invention is particularly suitable for injection and exhibits long-lasting efficacy and excellent stability of the drug.
  • the SECl mutant protein as described hereinbefore, is an active ingredient exhibiting excellent effects on prevention, symptom alleviation and treatment of mastitis of dairy cows, via an improved immune function of lactating or non-lactating dairy cows, and can be prepared by various methods known in the art.
  • the content of the active ingredient as defined above, is in the range of 0.001 to 50% by weight, based on the weight of solid microparticles. Where the content of the active ingredient is too low, it is difficult to exert pharmacological effects thereof. In contrast, where the content of the active ingredient is too high, it may cause occurrence of aggregation and denaturation thereof in water-insoluble solvents. More preferably, the content of the active ingredient is in the range of 0.01 to 20% by weight.
  • the formulation in accordance with the present invention contains various specific ingredients.
  • the protein-stabilizing excipient is an ingredient which enables formation of the active ingredient SECl mutant protein into particles while maintaining stability thereof.
  • the present inventors have selected feasible excipient candidates from a various kinds of excipients known to have protein stabilizing effects and have carried out confirmation experiments on whether these excipient candidates have effects on formation of solid microparticles and protein stabilization. Taking into consideration problems exhibited by spray drying micro- granulation, a microgranulation process was carried out via lyophilization.
  • TABLE 1 shows whether solid microparticles are formed or not when lyophilizing a mixture of the SECl mutant protein and excipients, and experimental results on percentage change in protein purity when the formulation containing such solid microparticles dispersed in oil was stored under room temperature conditions (25°C, 60% RH) and under severe conditions (40°C, 75% RH) for 4 weeks, respectively.
  • TABLE 1 also shows the results obtained when the water-soluble SECl mutant protein alone was dispersed in oil.
  • examples of the preferred protein-stabilizing excipients that can be used in the formulation of the present invention include, but are not limited to, sodium chloride, polyethyleneglycol (for example, PEG 8000), dis- accharides (for example, lactose, maltose and sucrose), glucose, tetramethylglucose, Pluronic (a triblock copolymer) and any combination thereof.
  • polyethyleneglycol is more preferable and a mixture of polyethyleneglycol and sodium chloride is particularly preferable.
  • the content of protein-stabilizing excipient is in the range of
  • the content of the excipient is in the range of 30 to 60% by weight.
  • the carbohydrate-based auxiliary excipient serves to maximize im- munopotency of the active ingredient SECl mutant protein while assisting action of the protein-stabilizing excipient.
  • the carbohydrate-based auxiliary excipients utilizable in the present invention include, but are not limited to, sodium carboxymethylcellulose, carboxymethylcellulose, hydroxypropylcellulose, chitosan, alginate, xylose, galactose, fructose, saccharose, dextran, chondroitin sulfate and any combination thereof.
  • particularly preferred is carboxymethyl cellulose.
  • the content of the carbohydrate-based auxiliary excipient is in the range of 0.1 to 90% by weight, based on the weight of solid microparticles. Where the content of the auxiliary excipient is too low, it is difficult to exert effects due to addition thereof. In contrast, where the content of the auxiliary excipient is too high, this may lead to failure of formation of solid particles during a lyophilization process. More preferably, the content of the auxiliary excipient is in the range of 0.5 to 50% by weight.
  • lipophilic materials may be added to the formulation in accordance with the present invention.
  • the lipophilic materials serve to improve dispersibility of microparticles containing the active ingredient SECl mutant protein, thereby improving injectability of the formulation.
  • examples of the lipophilic materials that can be used in the present invention include, but are not limited to, phosphatidylserine, phosphatidylethanolamine, lecithin, phosphatidylcholine-based materials (for example, stearoyl phosphatidylcholine and arachidonyl phosphatidylcholine), myristic acid, palmitic acid, stearic acid, sorbitan monooleate, polysorbate, glyceryl stearate, sorbitan palmitate, sorbitan stearate and any combination thereof.
  • Particularly preferred are phosphatidylcholine-based materials.
  • the content of the lipophilic material is in the range of 0.1 to 10% by weight, based on the weight of solid microparticles. Where the content of the lipophilic material is too low, it is difficult to sufficiently exert addition effects thereof. In contrast, where the content of the lipophilic material is too high, this may lead to failure of formation of solid particles after completion of lyophilization.
  • the content of the lipophilic material is preferably in the range of 0.1 to 5% by weight.
  • the biocompatible oils that can be used in the present invention preferably include, but are not limited to, edible oil, mineral oil, squalene, squalane, mono-, di- and triglyceride, and any combination thereof.
  • edible oils include soybean oil, corn oil, olive oil, safflower oil, cottonseed oil, peanut oil, sesame oil and sunflower oil. Particularly preferred is soybean oil.
  • the fatty acid ester-based compound preferably include, but is not limited to, monoglyceride, diglyceride, triglyceride, isopropylpalmitate, isopropylmyristate, benzoic acid, ethyl linoleate and any combination thereof. Particularly preferred is isopropylmyristate.
  • biocompatible oils and fatty acid ester-based compounds may be used, alone or in combination. Combined use thereof as the dispersion medium is more preferable in terms of improved injectability and maximized dispersion effects. This fact can also be confirmed from the results of Experimental Example 3 which will be illustrated hereinafter.
  • biocompatible oils and fatty acid ester-based compounds combined use of soybean oil and isopropylmyristate provides better injectability of the formulation.
  • the content of the biocompatible oil may be, for example, in the range of 1 to 99% by weight, based on the total weight of the dispersion medium.
  • the content thereof is in particular preferably in the range of 20 to 40% by weight.
  • An amount of microparticles added relative to the dispersion medium may be determined taking into consideration an optimal single-injection dose, injectability of the dispersion and the like and is preferably in the range of 1 to 99% by volume on the basis of the total volume. If necessary, it is possible to use the formulation in which the above dispersion was re-dispersed in physiological saline.
  • a method for preparing an SECl mutant protein formulation comprising: [33] (a) mixing an SECl mutant protein, a protein-stabilizing excipient, a carbohydrate- based auxiliary excipient and a lipophilic material;
  • step (b) solid microparticles are fabricated to have a particle diameter of about 5 to 200 D. Where the particle diameter is too small, aggregation of microparticles occurs, thus making it difficult to achieve sufficient dispersion and leading to deterioration of sustained-release properties of the active ingredient. Conversely, where the particle diameter is too large, precipitation of microparticles occurs in the dispersion medium, thus undesirably making it difficult to maintain the dispersed state.
  • FIG. 1 is a graph showing results of determination on ⁇ -IFN levels in blood collected after injection of formulations of Examples and Comparative Examples into mice, respectively, using a mouse cytokine ELISA kit;
  • FIG. 2 is a graph showing results of determination on changes in the number of somatic cells in milk collected prior to administration, and 2, 4, 6 and 10 weeks post administration, a total of five times, following injection of a formulation of Example 1 into lactating dairy cows having more than 510 somatic cells/ml of milk;
  • FIG. 3 is a graph showing results of determination on the number of somatic cells in milk collected after injection of a formulation of Example 1 and Lavac StaphTM ( Staphylococcus Aureus Bacterin)(Boehringer Ingelheim) into lactating dairy cows having more than 5x10 somatic cells/ml of milk, respectively.
  • Lavac StaphTM Staphylococcus Aureus Bacterin
  • SECl mutant protein formulations were prepared according to the following formula given in TABLE 2 below.
  • an SECl mutant protein, sodium chloride, carboxymethylcellulose and phosphatidylcholine were mixed together, the resulting mixture was lyophilized to prepare solid microparticles having an average particle diameter of about 50 to 80 D, and the solid microparticles were dispersed in soybean oil, thereby preparing a desired formulation.
  • mice were carried out using mice as follows. Specifically, each formulation was added to soybean oil such that a concentration of the SECl mutant protein was diluted to 40 D. The diluted formulations were intraperitoneally injected into 4-week old, male Balb/c mice and blood was collected 0, 2, 4, 8, 16 and 24 days post-administration. Thereafter, ⁇ -IFN levels in blood thus collected were determined using a mouse cytokine ELISA kit. The results thus obtained are shown in FIG. 1.
  • TABLE 5 presents protein contents determined when formulations, prepared by lyophilizing a mixture of an SECl mutant protein and excipients to obtain solid mi- croparticles and dispersing the solid microparticles in oil, were stored under room temperature conditions (25°C, 60% RH) and under severe conditions (40°C, 75% RH) for 24 weeks, respectively.
  • room temperature conditions 25°C, 60% RH
  • severe conditions 40°C, 75% RH
  • Example 1 As can be seen from TABLE 5, the formulation of Example 1 exhibited stable results without changes in protein contents for 24 weeks under room temperature conditions and under severe conditions, while the formulations of Example 7 through 10 exhibited a tendency of decreases in protein contents.
  • these results represent that the content of sodium chloride constituting solid microparticles of SECl mutant protein affects stability of the protein. Therefore, it can be seen that the particularly preferred content of sodium chloride is less than 60% by weight when sodium chloride is used as the protein-stabilizing excipient. Nonetheless, the above experimental results have confirmed that formulations of the present invention including the formulations of Example 7 through 10 generally ensure excellent stability of the SECl mutant protein even when they are stored under severe conditions (40°C, 75% RH) for a prolonged period of time (24 weeks).
  • mice 14 days after the first, second and third administration, respectively, blood was collected from mice (10 animals/ administration) followed by isolation of sera, and the titer of antibody specific for SECl mutant protein was analyzed using peroxidase-conjugated goat anti-mouse IgG (ICN. #55550). The results thus obtained are given in TABLE 6 below.
  • Example 1 As can be seen from TABLE 6, the formulation of Example 1 exhibited excellent antibody-producing ability with respect to contents of the SECl mutant protein in mice in vivo.
  • Example 1 As a somatic experiment of subject animals in order to verify immunopotentiating effects in dairy cows, a formulation of Example 1 was administered to 295 lactating dairy cows having more than 5x10 somatic cells/ml of milk via intramuscular injection and milk was collected 0, 2, 4, 6 and 10 weeks after administration of the formulation, a total of five times. Changes in the number of somatic cells in the collected milk were measured. The results thus obtained are shown in FIG. 2.
  • Example 1 As can be seen from FIG. 2, the formulation of Example 1 has continuously exhibited reduction effects of somatic cells in milk, starting from 4 weeks of administration up to 10 weeks.
  • This example is a somatic experiment of subject animals for comparison and verification of immunopotentiating effects of a formulation of Example 1 in dairy cows.
  • Lavac StaphTM a Staphylococcus aureus vaccine against mastitis in dairy cows (available from Boehringer Ingelheim)
  • Experiment was carried out using 295 lactating dairy cows having more than 5x10 5 somatic cells/ml of milk.
  • the formulation of Example 1 was intramuscularly injected into 278 dairy cows and the comparative formulation was intramuscularly injected into 17 dairy cows. Thereafter, milk was collected and the number of somatic cells in the milk was measured. The results thus obtained are shown in FIG. 3.
  • Example 1 As can be seen from FIG. 3, the formulation of Example 1 has exhibited better results in a reduction rate of somatic cells in milk, as compared to Lavac Staph of Comparative Example.
  • Microparticles of an SECl mutant protein were prepared by means of a lyophilization method having the most ideal drying temperature (eutectic point) conditions under which stability of the SECl mutant protein is maintained with formation of microparticles, and a spray drying method disclosed in Korean Patent No. 359252, respectively. Experimental conditions and the results thus obtained are given in TABLE 7.
  • yield (%) of microparticles by the spray drying method was about 11%
  • yield (%) of microparticles by the lyophilization method in accordance with the present invention was about 99%, thus representing a significant difference therebetween. That is, in producing the SECl mutant protein, it can be seen that preparation of SECl mutant protein microparticles via lyophilization is only suitable for mass production, thus making it possible to enter commercialization.
  • a formulation containing an SECl mutant protein in accordance with the present invention is capable of achieving effective in vivo delivery of a water-soluble mutant protein while maintaining activity thereof by inclusion of a protein-stabilizing excipient, a carbohydrate-based auxiliary excipient, a lipophilic material and a dispersion medium.
  • the formulation in accordance with the present invention exhibits excellent effects on prevention and treatment of mastitis of dairy cows via an enhanced immunopotentiating effects due to superior antibody-producing ability when administered to dairy cows.
  • the formulation of the present invention can also be used as an injectable preparation due to excellent injectability.

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Abstract

L'invention concerne une formulation d'une protéine mutante SECI, une toxine de Staphylococcus aureus, présentant d'excellents effets sur la prévention, l'atténuation du symptôme et le traitement de la mammite, via une fonction immune améliorée de lactation ou de non-lactation de vaches laitières, comprenant des microparticules solides contenant la protéine mutante SECI comme ingrédient actif, un excipient de stabilisation de protéine, un excipient auxiliaire à base de carbohydrate et un matériau lipophile, dispersés dans un composé à base d'huile biocompatible et/ou d'un ester d'acide gras biocompatible. La formulation selon l'invention fournit une production in vivo efficace d'une protéine mutante hydrosoluble, laquelle est un ingrédient actif, tout en conservant l'activité de celui-ci. En outre, la formulation selon l'invention présente d'excellents effets thérapeutiques sur la prévention et le traitement de la mammite, via des effets d'immunopotentialisation accrus, en raison d'une aptitude élevée à produire des anticorps lorsque ladite formulation est administrée aux vaches laitières. En outre, la formulation selon l'invention peut être utilisée comme préparation injectable, du fait de son excellente injectabilité.
PCT/KR2005/003871 2004-12-01 2005-11-16 Formulation de proteine mutee sec1, et procede de formulation de celle-ci WO2006059846A1 (fr)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8889150B2 (en) 2010-03-17 2014-11-18 SOCPRA—Sciences et Génie, s.e.c. Bacterial vaccine components from Staphylococcus aureus and uses thereof
EP3802573A4 (fr) * 2018-06-08 2022-04-27 Republic of Korea (Animal and Plant Quarantine Agency) Composition de vaccin comprenant une protéine recombinée d'entérotoxine atténuée et de cytotoxine de staphylococcus aureus
US11324815B2 (en) 2016-10-21 2022-05-10 Socpra—Sciences et Genie, S.E.C. Vaccine constructs and uses thereof against Staphylococcus infections

Citations (3)

* Cited by examiner, † Cited by third party
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US8889150B2 (en) 2010-03-17 2014-11-18 SOCPRA—Sciences et Génie, s.e.c. Bacterial vaccine components from Staphylococcus aureus and uses thereof
US9566322B2 (en) 2010-03-17 2017-02-14 SOCPRA—Sciences et Génie, s.e.c. Bacterial vaccine components and uses thereof
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AR054710A1 (es) 2007-07-11
MY140503A (en) 2009-12-31

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