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    Process for the preparation of a new-type active substance selectively inhibiting food intake
    6.
    发明授权
    Process for the preparation of a new-type active substance selectively inhibiting food intake 失效
    制备选择性抑制食物摄入的新型活性物质的方法

    公开(公告)号：US4588685A

    公开(公告)日：1986-05-13

    申请号：US632439

    申请日：1984-07-19

    申请人： Jozsef Knoll ,  Janos Nagy ,  Huba Kalasz ,  Berta Knoll

    发明人： Jozsef Knoll ,  Janos Nagy ,  Huba Kalasz ,  Berta Knoll

    IPC分类号： A61K35/14 ,  A61K35/16 ,  A61K38/00 ,  A61K38/16 ,  A61P3/04 ,  C07K1/16 ,  C07K1/34 ,  C07K14/435 ,  C07K14/47 ,  C12P21/06

    CPC分类号： C07K14/4702 ,  A61K38/00 ,  Y10S530/83

    摘要： The invention involves isolating a food intake suppressant by subjecting blood serum to ultrafiltration transmitting up to a molecular weight of 30,000 daltons, partially evaporating the filtrate, removing the insoluble part from the concentrate obtained, adding trichloroacetic acid up to a concentration of 5 to 25 weight/volume percent to the liquid phase at a temperature of 0.degree. to 10.degree. C., removing the proteins precipitated, subjecting the obtained solution to chromatography on a gel with a void volume below a molecular weight of 4000 daltons, eluting with a solution of pH 6.0 to 7.0, concentrating the biologically active fractions, chromatographing again on a gel with a void volume below a molecular weight of 4000 daltons, fractionating by elution with water, lyophilizing the active fractions, dissolving the lyophilized fractions in a buffer of pH 8.1 to 8.2, adding trypsin and chymotrypsin to the solution in a substantially identical amount of 0.01 to 0.2 by weight as calculated for the total weight of the lyophilized product, subjecting the mixture to digestion at a temperature of 36.degree. C. to 39.degree. C., preferably with occasionally shaking from 20 to 30 hours, adding after the first 5 hours trypsin and chymotrypsin in a substantially half amount as calculated for the original enzyme quantity and continuing the digestion, then adding trichloroacetic acid to the reaction mixture up to a concentration of 4 to 6 weight/volume percent at a temperature between 0.degree. and 10.degree. C., keeping the mixture at a temperature between -15.degree. C. and 5.degree. C. for 1 to 24 hours, removing the insoluble part, subjecting the reaction mixture to chromatography on a gel with a void volume below a molecular weight of 4000 daltons, fractionating by elution with water and lyophilizing the biologically active fractions.

    摘要翻译： 本发明涉及通过使血清进行超滤，分子量为30,000道尔顿，部分蒸发滤液，从所得浓缩物中除去不溶部分，加入高达5至25重量％的三氯乙酸，分离食物摄取抑制剂 /体积％至液相，在0℃至10℃的温度下，除去沉淀的蛋白质，将得到的溶液在孔隙体积低于4000道尔顿的凝胶上进行色谱分离，用 浓缩生物活性级分，再次在空白体积低于4000道尔顿的凝胶上进行色谱分离，用水洗脱分级，冻干活性级分，将冻干级分溶解在pH8.1缓冲液中至 8.2，将胰蛋白酶和胰凝乳蛋白酶以基本相同的量加入到溶液中，计算为0.01至0.2重量份 d，冻干产物的总重量，在36℃至39℃的温度下进行消化，优选在20至30小时内偶尔振荡，在第一个5小时后加入胰蛋白酶和胰凝乳蛋白酶 基本上为原始酶量计算的一半量并继续消化，然后在0℃和10℃之间的温度下，向反应混合物中加入高达4至6重量/体积百分数的浓度的三氯乙酸，保持混合物 在-15℃至5℃的温度下反应1至24小时，除去不溶部分，将反应混合物在空隙体积低于4000道尔顿的凝胶上进行色谱分离，通过用 水并冻干生物活性级分。

    Purified appetite-regulating substances of biological orgin, their antibodies, immunocomplexes of the appetite--regulating substances formed with the antibodies and processes for preparing same
    7.
    发明授权
    Purified appetite-regulating substances of biological orgin, their antibodies, immunocomplexes of the appetite--regulating substances formed with the antibodies and processes for preparing same 失效
    纯化的生物orgin的食欲调节物质，其抗体，与抗体形成的食欲调节物质的免疫复合物及其制备方法

    公开(公告)号：US4859766A

    公开(公告)日：1989-08-22

    申请号：US134403

    申请日：1987-12-17

    申请人： Jozsef Knoll ,  Sandor Harmath ,  Janos Nagy ,  Berta Knoll

    发明人： Jozsef Knoll ,  Sandor Harmath ,  Janos Nagy ,  Berta Knoll

    IPC分类号： A61K39/00 ,  A61K35/16 ,  A61K39/395 ,  A61P1/00 ,  C07K1/22 ,  C07K14/00 ,  C07K14/435 ,  C07K16/00

    CPC分类号： A61K35/16 ,  Y10S530/83

    摘要： The selective appetite-regulating substances satietin and satietin-D had up to the present been prepared from human or animal blood serum or plasma by ultrafiltration, gel chromatography, treatment with trichloroacetic acid, affinity chromatography and optionally treatment with a proteolytic enzyme.According to the invention, satietin and satietin-D are obtained in a purified form by using immunoabsorption. Being glycoproteins, both satietin and satietin-D form antibodies (antisatietin and antisatietin-D, respectively) in the living organism and can be isolated from the blood and bound to a gel column. For the preparation of satietin and satietin-D, respectively, the human or animal blood plasma or serum is subjected to ultrafiltration and the ultrafiltrate, containing in addition to satietin and satietin-D other constituents with a molecular weight below 50000 daltons, is contacted with the gel column. The antibody bound to the gel column forms with satietin or satietin-D, respectively, an immunocomplex remaining on the column while the other constituents of the ultrafiltrate can be eluted by a buffer. Then, the immunocomplex is cleaved, whereby satietin or satietin-D, respectively, can be eluted in a purified form from the column. The column containing the antibody may repeatedly be used.

    摘要翻译： 目前通过超滤，凝胶色谱，三氯乙酸处理，亲和层析和任选用蛋白水解酶处理，人体或动物血清或血浆中选择性食欲调节物质satietin和satietin-D已经制备。 根据本发明，通过使用免疫吸附法以纯化的形式获得了satietin和satietin-D。 作为糖蛋白，在生物体中，satietin和satietin-D分别形成抗体（抗血小板素和抗血小板素-D），并且可以从血液中分离并结合到凝胶柱。 为了分别制备satietin和satietin-D，将人或动物血浆或血清进行超滤，并且除了含有分子量低于50000道尔顿的satietin和satietin-D其它成分之外，还含有超滤液与 凝胶柱。 与凝胶柱结合的抗体分别与satietin或satietin-D形成，残留在柱上的免疫复合物，而超滤液的其它成分可被缓冲液洗脱。 然后，免疫复合物被切割，从而可以从柱中以纯化形式洗脱satietin或satietin-D。 可以重复使用含有抗体的柱。