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    • 专利摘要:
    1502348 Bacterial extracts from corynebacteria AGENCE NATIONALE DE VALORISATION DE LA RECHERCHE 5 May 1975 [6 May 1974] 18752/75 Heading A5B Acetylated water soluble extracts of molecular weight in the range 1000 + 200 to 8000 Π200 and containing diaminopimelic acid (DAP) are obtained from corynebacteria cells, e.g. corynebacterium parvum, by acetylating delipidated bacterial residues of the cells containing DAP, e.g. with a mixture of acetic anhydride and pyridene, and isolating the water soluble extracts e.g. by ethanol/water extraction. The acetylated water soluble extracts may be further purified and separated by filtering on a polyacrylamide gel or by chromatography on an adsorbent. The purified acetylated extracts obtained are essentially mixtures of acetylated water soluble fragments of the cell wall comprising disaccharide tetrapeptide, tetrasaccharide heptapeptide and its dimer, trimer and tetramer. The extracts in combination with a diluent can be used as a pharmaceutical composition for promoting resistance to viral or bacterial infections.
    公开号:SU710503A3
    申请号:SU752131286
    申请日:1975-05-05
    公开日:1980-01-15
    发明作者:Жолль Пьер；Миглиор-Самур Даниель
    申请人:Ажанс Насьональ Де Валоризасьон Де Ля Решерш (Анвар) (Фирма)；
    IPC主号:
    专利说明:

    The invention relates to the pharmaceutical industry, to the production of drugs used in medicine.
    A known method of producing acetylated water-soluble extracts having an immunostimulating effect by treating low-fat bacterial residues with a mixture of pyridine and acetic anhydride, followed by extraction of the target product with ethanol, water, isolation and purification by physicochemical methods of gi.
    However, the known cno & oR ‘does not allow to obtain an extract with high immunostimulating properties.
    The aim of the invention is to enhance the immunostimulating properties of th resistance to infections of viral origin.
    This goal is achieved by the fact that as a starting material using pseudodiphtheria coli SocietietKJcteHurn rag ^ itt The method is as follows.
    Cells of pseudodiphtheria bacilli are first subjected to degreasing, then acetylated by treatment with acetic anhydride in the presence of pyridine.
    Then water-soluble extracts are isolated by extraction with alcohol, followed by extraction with water.
    Water-soluble extracts are isolated by treatment with alcohol (ethanol), the reaction medium obtained after acetylation, the insoluble part is separated by centrifugation, the alcohol solution is concentrated.
    Then, the residue obtained is treated with distilled water, the water-insoluble fractions are separated by centrifugation, and the upper aqueous layer is lyophilized, which contains acetylated water-soluble extracts that are simultaneously soluble in an alcohol-pyridine mixture and in water.
    As an alcohol – pyridine mixture, a mixture of ethanol and pyridine (approximately in a ratio of 10: 1) by volume is used. All centrifugation steps are carried out at low temperature, preferably at 0-6 ° C.
    Water-soluble extracts can be purified and isolated by appropriate physicochemical methods "·
    Λ * ’= ,.
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    ZD / ΐ. 710503, namely, the chromatoGRF ^ IJSCH ^^ J ;.
    bents such as A ’/’ or by filtration, for example, through a polyacrylamide gel using B-iopet Ryu ,. BiocreE P6 or B-ioget P4.
    The crude '(crude) water-soluble extract obtained after Filtering, then chromatography, eluted with solvents such as diluted or acetic acid. the pyridine-acetic acid mixture is formed by a combination of acetylated substances, including acetylated disaccharide-tetrapeptide, acetylated tetrasaccharide-heptapeptide, as well as its dimers, trimers and tetramers, in some cases associated with non-aminated acetylated reducing sugars. In the process of filtration and chromatography, water-soluble substances are released in order of increasing mole. ocular weights.
    The activity of water-soluble extracts is associated with the presence of diaminopimelic acid,
    Acetylated water-soluble extracts obtained by the proposed method have enhanced immunostimulating properties and resistance to infections of viral origin.
    So in guinea pig, water-soluble extracts cause an increase in the percentage of antibodies at doses higher or equal to 0.1 mg subcutaneously. Water-soluble extracts, together with additives, such as antibiotics, can be used as anti-infusion agents. In these compositions, the percentage of acetylated water-soluble extracts is usually higher than 0.1%.
    The compositions may be administered orally, rectally, parenterally or in the form of aerosols.
    For an adult, doses are 10-50 mg per day.
    Example 1. 20 g of bacterial defatted residues obtained from Cotynebacterium parvuni cells are stirred for 66 hours at 28 ° C in 500 cm 3 of acetic anhydride pyridine mixture (2-3 by volume). Then add 5000 mix 5 part separate (4000 rpm). The research institutes of tilled water are dried to water and part is separated by centrifugation (4000 rpm). The top layer is lyophilized. Thus, 3.280 g of crude acetylated water-soluble extract is obtained.
    TsNIIIPI Order 8785/53 cm 3 of ethanol, then another hour. Insoluble by centrifugation. Top layer, under reduced pressure. Residue is recovered. 100 cm 3 insoluble in
    Example 2. The acetylated water-soluble crude extract obtained in example 1, is purified by filtering on a column with Vkz ^ eEPYu, collecting fractions of 1.5 cm 3 No. 1-500, and the elution is carried out using 0.1 N. acetic acid and then optically controlled on a spectrophotometer at 220 and 280 nm. Fractions No. 293-340 are combined and lyophilized. Thus, 0.195 g of a water-soluble extract is obtained, which is purified by chromatography on ΌΕ AB-A balanced with a 0.01 M pyridine solution with a pH of 7.7. Eluting with a 0.01 M pyridine solution, 0.110 g of a substance enriched with diaminopimelic acid is obtained, then, eluting with a solvent gradient of 0.01 to 0.2 M pyridine solution with a pH of 7.7 to 5.5, 0.080 g of acetylated water-soluble is obtained. extract.
    The water-soluble extract is a yellowish powder, includes the following amino acids: alanine, glutamic acid, diaminopimelic acid, aspartic acid and glycine; aminosugar: N — acetylglucosamine, N-acetylmuramic acid; reducing non-laminated sugars: mannose, glucose and trace of galactose. Molecular weight (counting on the base with 3 residues of alanine in the molecule) 3750 + 250.
    The percentage of amino acids 35 lot 22 + 3%; amino sugars 26 + 2%.
    The proposed method allows to obtain acetylated water-soluble extracts that have enhanced immunostimulating properties and resistance to infections of viral origin and can be used in human therapy or in veterinary medicine.
    权利要求:
    Claims (1)
    [1]
    Claim
    A method for producing acetylated water-soluble extracts having an immunostimulating effect by treating low-fat bacterial residues with a mixture of pyridine and acetic anhydride. With the subsequent extraction of the target product with ethanol, water, isolation and purification by physicochemical methods, it is necessary that, in order to enhance the immunostimulating properties and resistance to infections of viral origin, CorMnebacteNuni par ^ pseudodiphtheria bacilli are used as starting material um
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    同族专利:
    公开号 | 公开日
    DE2519937A1|1975-11-27|
    BE828636A|1975-10-30|
    IL47207A|1978-03-10|
    CH600885A5|1978-06-30|
    HU171392B|1978-01-28|
    JPS50155686A|1975-12-16|
    FR2269962A1|1975-12-05|
    CA1043776A|1978-12-05|
    IL47207D0|1975-06-25|
    DK195075A|1975-11-07|
    US4014992A|1977-03-29|
    GB1502348A|1978-03-01|
    NL7505247A|1975-11-10|
    SE7505033L|1975-11-07|
    FR2269962B1|1978-02-03|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    FR2321896B1|1975-08-29|1978-08-11|Anvar|
    DE3011461C2|1980-03-25|1986-10-30|Dr. Madaus & Co, 5000 Köln|Use of propionibacteria|
    JPS57163835U|1981-04-07|1982-10-15|
    JPS6028999A|1983-06-30|1985-02-14|Maruho Kk|Protein having cell proliferation accelerating action, its composition and its preparation|
    US4663306A|1983-09-23|1987-05-05|Ribi Immunochem Research, Inc.|Pyridine-soluble extract-refined detoxified endotoxin composition and use|
    US5976580A|1995-06-07|1999-11-02|Novus International, Inc.|Nutrient formulation and process for enhancing the health, livability, cumulative weight gain or feed efficiency in poultry and other animals|
    JPH11171783A|1997-08-15|1999-06-29|Vetoquinol Sa|Oral administration of bacteria at concentration that causes cell-mediated immunity and body weight increase in specific animal|
    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    FR7415571A|FR2269962B1|1974-05-06|1974-05-06|
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