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The preparation of exosomes - ONKOSOM, its production method, a method of preventing malignancies and solid tumors immunotherapy method for using it

机译:外泌体的制备-ONKOSOM,其生产方法,预防恶性肿瘤的方法和实体瘤免疫疗法的使用方法

摘要

1. Preparation of exosomes isolated culture K562 / i-S9 cells containing an enzyme immunoassay according to the protein in an amount of 0.9-1.5 g / 10kletok, CEA antigens in an amount of 0,045-0,075 mcg / 5T4 and 10kletok -0,03- 0.05 ug / 10kletok and ligands FasL- less than 0.005 g / 10kletok and TRAIL - less than 0.004 g / 10kletok.2. A method for producing a preparation according to claim exosomes. 1, which consists in the fact that the cell line K562 / i-S9 cultured at 36,5-37,5 ° C for 36-72 hours and maintained at 43-44 ° C, regenerated with 36 , 5-37,5 ° C for 3.5-4.5 hours, frozen and homogenized to a powder, ultraviolet ray is irradiated homogenate and precipitated cell fragments by centrifugation, the resulting supernatant was added ethyl alcohol to give its concentration of 84 ° and centrifuged at 60000g for 20-25 minutes, the resulting precipitate was dissolved in distilled water Is subjected to ultrafiltration and liofilizatsii.3. A method for preventing malignancy in a human vaccine comprising a combination of a preparation of exosomes according to Claim. 1 and P-type oligodeoxynucleotide constituting GGGGCGCCGTGATGGCGAGCGCGCC sequence of nucleotides that is administered s.c. 1 time per year in an effective amount, preferably in doses of 0.01-0.04 mg / kg kazhdyy.4. A method of immunotherapy of solid tumors, comprising a patient vaccination combination exosome preparation according to Claim. 1 and oligodeoxynucleotide P-type, which is a sequence GGGGCGCCGTGATGGCGAGCGCGCC nucleotides which is administered subcutaneously for at least 2 months, at least 1 time per week in an effective amount, preferably in doses of 0.01-0.04 mg / kg kazhdyy.5. . A method according to claim 4, characterized in that during at least part of the period of vaccination orally administered drug - reverse transcriptase inhibitor in effective doses.
机译:1.外泌体分离培养K562 / i-S9细胞的制备,该细胞包含根据蛋白质含量为0.9-1.5 g / 10kletok的酶免疫测定,含量为0.045-0.075 mcg / 5T4和10kletok -0.03的CEA抗原-0.05 ug / 10kletok和配体FasL-小于0.005 g / 10kletok和TRAIL-小于0.004 g / 10kletok.2。 13.根据权利要求1的外来体的制备方法。 1,其事实在于,细胞系K562 / i-S9在36,5-37,5°C下培养36-72小时并保持在43-44°C,并在36,5-37,5下再生于3.5°C-4.5小时,冷冻并匀化为粉末,通过离心辐照紫外线以匀浆并沉淀细胞碎片,向所得上清液中加入乙醇使其浓度为84°C,并在60000g下离心20-25分钟然后,将得到的沉淀物溶解在蒸馏水中,进行超滤和硅藻土处理。3。预防人疫苗中恶性肿瘤的方法,其包括根据权利要求1的外泌体制剂的组合。 1和P型寡聚脱氧核苷酸构成GGCGCGCCGTGATGGCGAGCGCGCC的核苷酸序列s.c.每年1次有效量,最好以0.01-0.04 mg / kg kazhdyy.4的剂量服用。实体瘤的免疫治疗方法,包括根据权利要求1的患者疫苗接种组合外来体制剂。 1和寡聚脱氧核苷酸P型,其是序列GGGGCGCCGTGATGGCGAGCGCGCC核苷酸,其以有效量,优选以0.01-0.04mg / kg kazhdyy.5的剂量皮下施用至少2个月,每周至少1次。 。 5.根据权利要求4所述的方法,其特征在于,在疫苗接种的至少一部分期间内,口服给药的药物-有效剂量的逆转录酶抑制剂。

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