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CN-121971500-A - A high content extract of saikosaponin B2, its preparation method and application, and a pharmaceutical composition containing the extract

CN121971500ACN 121971500 ACN121971500 ACN 121971500ACN-121971500-A

Abstract

The invention provides a high-content extract of saikosaponin B2, a preparation method and application thereof, and a combined drug containing the extract, belonging to the technical field of natural drug preparation. According to the invention, the saikosaponin A and saikosaponin D in the bupleurum raw material are efficiently extracted by an alkaline ethanol solution, the saikosaponin A and saikosaponin D are further efficiently split by macroporous adsorption resin, the saikosaponin D is quantitatively converted into saikosaponin B2 by a chemical method, and the high-content effective part of saikosaponin B2 is obtained. The high content effective part of the saikosaponin B2 contains only a small amount of saikosaponin A except the main component saikosaponin B2, thereby greatly reducing toxicity while ensuring the bioactivity. The high-content effective part of the saikosaponin B2 can be used for preparing medicines for treating and preventing diseases such as gout, rheumatoid arthritis, acute enteritis, skin photodamage, skin inflammation and the like, and can be used independently or combined with other medicines.

Inventors

  • LI FU
  • WANG FEI
  • Xi Ruiying
  • JI MINGHUI

Assignees

  • 中国科学院成都生物研究所

Dates

Publication Date
20260505
Application Date
20260408
Priority Date
20250410

Claims (10)

  1. 1. The high-content extract of saikosaponin B2 is characterized in that the preparation method comprises the following steps: (1) Crushing a bupleurum raw material, and adding a solvent for extraction to obtain bupleurum extracting solution, wherein the solvent is 10% -90% ethanol water solution with pH value of 8-14; (2) Concentrating the bupleurum extracting solution, regulating the pH value to 2-7 by using acid, sequentially eluting macroporous adsorption resin columns by using an ethanol aqueous solution 1, an ethanol aqueous solution 2 and an ethanol aqueous solution 3 through macroporous adsorption resin, collecting eluent of the ethanol aqueous solution 3, and concentrating to obtain concentrated solution, wherein the ethanol concentration in the ethanol aqueous solution 1, the ethanol aqueous solution 2 and the ethanol aqueous solution 3 is 10% -90%, and the ethanol concentration in the ethanol aqueous solution 1, the ethanol aqueous solution 2 and the ethanol concentration in the ethanol aqueous solution 3 are sequentially increased; (3) Adding acid into the concentrated solution to adjust the pH to 1-6, and then reacting to obtain the extract with high content of saikosaponin B2, wherein the content of saikosaponin B2 in the extract with high content of saikosaponin B2 is more than 60%.
  2. 2. The extract of claim 1, wherein in the step (1), the bupleurum raw material is bupleurum crude drug, bupleurum preparation or bupleurum decoction piece, the bupleurum is dry root of umbrella-shaped plant bupleurum Bupleurum chinense DC or bupleurum angustifolium Bupleurum scorzonerifolium Willd, the solvent is 60% -85% ethanol aqueous solution with pH value of 10-13, the mass ratio of the bupleurum raw material to the solvent is 1:2-20, the extraction temperature is 20-100 ℃, the extraction times are 1-5 times, and the extraction time is 1-24 hours each time; In the step (2), the ethanol content is 20% -48% after the bupleurum extract is concentrated, the pH is 4-6.5 after the bupleurum extract is regulated by acid, the types of macroporous adsorption resin are AB-8, D-101, MQ-10, NKA series, HPD series, ADS series, DM series, T series, LX series or AXAB series, and the ethanol concentration in the ethanol aqueous solution 1, the ethanol aqueous solution 2 and the ethanol aqueous solution 3 is 20% -48%, 48% -65% and 65% -90% in sequence; In the step (3), after acid addition and adjustment, the pH is 4-6, the reaction temperature is 30-110 ℃ and the reaction time is 0.5-10 hours.
  3. 3. The extract according to claim 2, wherein in the step (1), the bupleurum processed product is vinegar-processed bupleurum, the solvent is a 70% -80% ethanol aqueous solution with a ph=11-12, the mass ratio of bupleurum raw material to solvent is 1:6-8, the extraction temperature is 60-80 ℃, the extraction times are 1-3 times, and the extraction time is 3-4 hours each time; in the step (2), the ethanol content is 35% -45% after the bupleurum extract is concentrated, the pH is 5-6 after the bupleurum extract is regulated by acid, the type of macroporous adsorption resin is AB-8, D-101, NKA-9, MQ-10 or HPD-100, and the ethanol concentration in the ethanol aqueous solution 1, the ethanol aqueous solution 2 and the ethanol aqueous solution 3 is 35% -45%, 50% -55% and 75% -80% in sequence; In the step (3), after acid addition and adjustment, the pH is 5, the reaction temperature is 80-100 ℃ and the reaction time is 3-4 hours.
  4. 4. A process for preparing an extract having a high content of saikosaponin B2 according to any one of claims 1-3, characterized in that it comprises the following steps: (1) Crushing a bupleurum raw material, and adding a solvent for extraction to obtain bupleurum extracting solution, wherein the solvent is 10% -90% ethanol water solution with pH value of 8-14; (2) Concentrating the bupleurum extracting solution, regulating the pH value to 2-7 by using acid, sequentially eluting macroporous adsorption resin columns by using an ethanol aqueous solution 1, an ethanol aqueous solution 2 and an ethanol aqueous solution 3 through macroporous adsorption resin, collecting eluent of the ethanol aqueous solution 3, and concentrating to obtain concentrated solution, wherein the ethanol concentration in the ethanol aqueous solution 1, the ethanol aqueous solution 2 and the ethanol aqueous solution 3 is 10% -90%, and the ethanol concentration in the ethanol aqueous solution 1, the ethanol aqueous solution 2 and the ethanol concentration in the ethanol aqueous solution 3 are sequentially increased; (3) And adding acid into the concentrated solution to adjust the pH to 1-6, and then reacting to obtain the extract with high content of saikosaponin B2.
  5. 5. Use of the high content extract of saikosaponin B2 according to any of claims 1-3 for the preparation of a medicament for the prevention and/or treatment of gout, gouty arthritis or rheumatoid arthritis.
  6. 6. The use according to claim 5, wherein the gout is acute gout.
  7. 7. The use according to claim 5, wherein the gouty arthritis is acute gouty arthritis.
  8. 8. Use of the high content extract of saikosaponin B2 according to any of claims 1-3 for the preparation of a medicament for the prevention and/or treatment of inflammatory bowel disease, peritonitis, diabetes, acute enteritis, skin photodamage, skin inflammation.
  9. 9. The use according to claim 8, wherein the inflammatory bowel disease is ulcerative colitis.
  10. 10. A combination comprising an extract of the high content of saikosaponin B2 according to any of claims 1-3 and a further drug, which is colchicine tablet or indomethacin, for simultaneous or separate administration of the same or different specification unit preparations.

Description

A high content extract of saikosaponin B2, its preparation method and application, and a pharmaceutical composition containing the extract Technical Field The invention belongs to the technical field of natural medicine preparation, and relates to a high-content extract of saikosaponin B2, a preparation method and application thereof, and a combined medicine containing the extract. Background Uric acid forms crystals at joints, kidneys and other parts to cause gout, and gout patients often have a plurality of complications, including kidney diseases, hypertension, diabetes mellitus, cardiovascular diseases and the like, and gout is a second most metabolic disease in China next to diabetes mellitus. At present, the first-line medicine in the acute onset stage of gout is colchicine or a non-steroidal anti-inflammatory medicine, but the defects of small safety range, easy medicine resistance, large toxic and side effects and the like generally exist, and the second-line medicine is glucocorticoid accompanied with joint and bone injury, so no better treatment means is recognized for gout at present. Radix bupleuri is the dry root of Umbelliferae plant Bupleurum chinense Bupleurum (known as "North Bupleurum chinense") or Bupleurum scorzonerifolium Bupleurum scorzonerifoliumWilld (known as "Nangunyi"). Radix bupleuri has effects of anti-inflammatory, protecting liver, relieving fever, and relieving pain, and can be used for treating headache, menoxenia, dysmenorrhea, common cold, gastric ulcer, duodenal ulcer, bile reflux gastritis, etc. caused by stagnation of liver qi. The bupleurum contains chemical components such as saponin, flavone, lignan, phytosterol and the like, wherein the main active components are saponin compounds. The total content of saikosaponin A and saikosaponin D in bupleurum according to the 2020 edition of Chinese pharmacopoeia is not less than 0.30%, however, saikosaponin A and saikosaponin D are main active ingredients of bupleurum and are main ingredients of bupleurum which generate toxicity (see Li Xiaoyu and the like, the acute toxicity experimental study of saikosaponin a on mice, chinese medicine alert, 2014,11 (8): 449-452; li Xiaoyu and the like, the acute toxicity experimental study of saikosaponin D on mice, chinese medicine alert, 2014,11 (12): 705-708). Compared with the saikosaponin A and the saikosaponin D, the toxicity of the saikosaponin B2 is obviously weakened, and the saikosaponin B2 has the effects of resisting infection, allergy, virus, shock and the like. However, the chemical synthesis method for preparing the saikosaponin B2 has the problems that the chemical synthesis process of the saikosaponin B2 is complex, multi-step reaction and fine condition control are required, the synthesis cost is high, various chemical reagents and solvents are required to be used in the chemical synthesis process, the reagents are generally expensive and can cause environmental pollution, and the chemical synthesis yield of the saikosaponin B2 is low, so that the saikosaponin B2 is difficult to produce in a large scale. The method for extracting the saikosaponin B2 from the Chinese medicinal bupleurum has lower cost and environmental protection, can retain the active ingredients of natural products, and is more suitable for industrial production. Therefore, the preparation of the bupleurum extract with high content of the saikosaponin B2 has important significance. However, the content of the saikosaponin B2 in the bupleurum crude drug is extremely low or even completely absent, and only a small amount of saikosaponin B exists in the vinegar-processed bupleurum, and is mainly formed by the conversion of saikosaponin D in the processing process. The method is characterized in that a total extract obtained by extracting bupleurum saponin B2 is subjected to macroporous resin purification to obtain a crude product of the bupleurum saponin B2, and the crude product of the bupleurum saponin B2 is prepared by silica gel column chromatography or recrystallization or a combination of the two methods, but the method has the following problems that (1) water and an alcohol solvent or a mixed solvent formed by a certain proportion is adopted as an extraction solvent, so that the extraction rate of the bupleurum saponin B2 in the extract is low, the yield of the bupleurum saponin B2 is between 0.24% and 0.44%, the total extract obtained by extracting the bupleurum saponin B2 is converted into the bupleurum saponin B2 in the extraction process, the subsequent macroporous resin method is used for realizing the effective separation of the bupleurum saponin B2 by the silica gel column chromatography or recrystallization or the combination of the two methods, and the method does not meet the requirements of high-quality purification of the bupleurum saponin B2 and the high-quality purification of the 3 when the crude product of the bupleurum saponin B2 is prepared by adopting water and the alcohol solven