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Use of extractive of BCG polysaccharide and nucleic acid for preparing medicine for treating viral skin diseases, and its injection and preparation method

A technology of BCG polysaccharide nucleic acid and extract, which can be used in skin diseases, medical formulas, drug combinations, etc., and can solve problems affecting daily life, redness, swelling, and pain at the injection site

Active Publication Date: 2015-03-04
HUNAN SIQI BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the human body, BCG bacterial proteins and miscellaneous bacteria are heterologous antigens or toxic substances, which can cause strong immune rejection and toxicity (because it needs to be used continuously for 3 months), which is manifested by the clinical use of this preparation Severe ulceration often occurs at the injection site of the patient, and about 5-10% of the patients have a fever at the same time, and a few patients have severe allergic reactions; (3) only the lower limit of the nucleic acid content is specified in the existing BCG polysaccharide nucleic acid injection , but in the long-term clinical application, it was found that when the nucleic acid content in each injection was greater than 110 μg, about 90% of the patients showed redness and swelling at the injection site, pain, and even severe swelling that affected daily life

Method used

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  • Use of extractive of BCG polysaccharide and nucleic acid for preparing medicine for treating viral skin diseases, and its injection and preparation method
  • Use of extractive of BCG polysaccharide and nucleic acid for preparing medicine for treating viral skin diseases, and its injection and preparation method
  • Use of extractive of BCG polysaccharide and nucleic acid for preparing medicine for treating viral skin diseases, and its injection and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment one BCG cultivation and harvesting

[0033] 1. Bacterial culture: Dissolve the strains preserved in liquid cryogenic temperature (BCG strain D2PB302 for the preparation of BCG in China, China National Institute for the Control of Pharmaceutical and Biological Products) at room temperature, inoculate in potato Sutong culture medium, and culture continuously at 37°C for 14-20 days ; or after continuous cultivation at 37°C for 15 days, transfer to improved liquid Sutong medium, and continuous cultivation at 37°C for 14-20 days.

[0034] Wherein, the preparation method of potato Sutong culture medium can be:

[0035] (1) Take and wash fresh potatoes (1), pierce them into cylinders with a piercer, and cut them into 4 cm long slopes with a knife; rinse the potato slopes with running drinking water for 1 hour, and then rinse the potato slopes with purified water; Wash the slanted block with Sutong medium, take 20ml of Sutong medium, and add it to a 100ml sterilize...

Embodiment 2

[0045] Embodiment two Preparation of BCG polysaccharide nucleic acid mixture

[0046] 1. Cell crushing and thermal phenol treatment: Add the collected cells to purified water at a ratio of 10:1, crush the cells with a tissue mash homogenizer (12000rpm / min), 3 min x 3 times, and pound the cells Then add 0.5-2.0 times the volume of hot phenol (30-100° C.) of the crushed bacteria suspension, and keep warm for 30 minutes to 1 hour while stirring at a low speed.

[0047] 2. Extraction of BCG polysaccharide nucleic acid mixture: Precipitate the hot phenolic mixture naturally for 1 to 10 days, absorb the supernatant, centrifuge at high speed in a tube centrifuge, and filter the supernatant through a 0.45um sterile filter to obtain BCG polysaccharide nucleic acid mixture.

Embodiment 3

[0048] Embodiment Three Preparation of Refined BCG Polysaccharide Nucleic Acid Extract

[0049] The K-PRIME 40II gel chromatography filtration system and GH-25 gel medium manufactured by MILLIPORE, USA were used to separate BCG polysaccharide nucleic acid, phenol and BCG protein. Concrete preparation process is as follows:

[0050] 1. Equilibrium: Use 0.9% normal saline to flow through the column at 1000mL / min in a forward direction until the conductance and pH of the column before and after the column are consistent and the balance process is completed.

[0051] 2. Sample loading: The BCG polysaccharide nucleic acid mixture is loaded through the sample loading system, and the sample loading speed is 600mL / min.

[0052] 3. Elution: After loading the sample, elute with 0.9% normal saline at a speed of 800mL / min, detect the effluent with a UV spectrometer (wavelength 260nm or 280nm), collect the target peak, and pass the collected target peak through 0.45 Filtered with a μm ...

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Abstract

The invention claims use of extractive of BCG polysaccharide and nucleic acid for preparing medicine for treating viral skin diseases. The invention also claims an injection of polysaccharide and nucleic acid comprising 0.32mg / ml-0.38mg / ml polysaccharide, 50µg / ml-100µg / ml nucleic acid, 0µg / ml-2.5µg / ml protein of BCG, and zero phenol and foreign bacteria. And the invention also claims a method for preparing the injection of BCG polysaccharide and nucleic acid with the extractive of BCG polysaccharide and nucleic acid.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals. Specifically, the present invention relates to the application of a BCG polysaccharide nucleic acid extract in the preparation of medicines for the treatment of viral skin diseases, and the BCG polysaccharide nucleic acid injection prepared from the BCG polysaccharide nucleic acid extract and its preparation method . Background technique [0002] Bacilli Calmette Guerin (BCG), a weakened strain of Mycobacterium bovis, is currently the most effective vaccine against tuberculosis. Since 1948, 3 billion people worldwide have been vaccinated, and its safety has been proven. In addition to being used for the prevention of tuberculosis, BCG can also be used for the treatment of various diseases, such as allergic diseases and tumors. However, due to its complex composition, the incidence of side effects such as rash and itching is relatively high. Therefore, purifying BCG and extracting its active ...

Claims

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Application Information

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IPC IPC(8): A61K31/715A61K9/08A61K47/42A61K35/74A61P17/00
CPCA61K31/715A61K9/0019A61K35/74A61P17/00
Inventor 宁云山关继峰
Owner HUNAN SIQI BIOPHARM
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