WO2022061637A1 - Procédé de détermination de la teneur en glycosaminoglycane fucosylé de faible poids moléculaire - Google Patents

Procédé de détermination de la teneur en glycosaminoglycane fucosylé de faible poids moléculaire Download PDF

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WO2022061637A1
WO2022061637A1 PCT/CN2020/117364 CN2020117364W WO2022061637A1 WO 2022061637 A1 WO2022061637 A1 WO 2022061637A1 CN 2020117364 W CN2020117364 W CN 2020117364W WO 2022061637 A1 WO2022061637 A1 WO 2022061637A1
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titrant
titration
volume
solution
optionally
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PCT/CN2020/117364
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English (en)
Chinese (zh)
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李振国
周剑波
务勇圣
张勇纯
王怡
王思瑶
韦怀福
郭亚芳
赵孝玉
徐春红
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牡丹江友搏药业有限责任公司
九芝堂股份有限公司
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Priority to PCT/CN2020/117364 priority Critical patent/WO2022061637A1/fr
Publication of WO2022061637A1 publication Critical patent/WO2022061637A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
    • G01N27/04—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance
    • G01N27/06—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance of a liquid
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
    • G01N31/16—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using titration
    • G01N31/18—Burettes specially adapted for titration

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  • the invention belongs to the technical field of medicine, and relates to a method for measuring the content of low-molecular-weight fucosified glycosaminoglycans, in particular to a method for measuring the content of low-molecular-weight fucosified glycosaminoglycans by volumetric method.
  • Fucosylated Glycosaminoglycan is a glycosaminoglycan with a special chemical structure and pharmacological activity that has only been found in echinoderms so far. It has a main chain similar to chondroitin sulfate and contains fucose. Fucosyl (Fucosyl, Fuc) side chain substituted glycosaminoglycan analogs (Yoshida et.al, Tetrahedron Lett, 1992, 33:4959-62; et.al, J Biol Chem, 1996, 271:23973-84).
  • LFG Low Molecular Weight Fucosylated Glycosaminoglycan, low molecular weight fucosylated glycosaminoglycan, hereinafter referred to as "LFG" is a glycosaminoglycan derivative obtained from sea cucumber body wall extraction and depolymerization. Lactose and glucuronic acid are polymerized. LFG is a polysaccharide drug. Due to the complex structure and polydispersity of the polysaccharide itself, quality control is difficult. Determination of sugar content is a necessary step in the research of polysaccharide drugs, and it is also an important part of quality control and product standard formulation.
  • polysaccharide drugs Due to the diverse sources of polysaccharide drugs and different structural types, there are many methods for the determination of polysaccharides. Common sugar groups in polysaccharide drugs include neutral hexose, pentose and deoxyhexose, negatively charged uronic acid, sialic acid, positively charged amino sugar, etc. Hydrolysis to monosaccharides establishes specificity for the assay.
  • LFG is a macromolecular polysaccharide substance, it is composed of a series of oligosaccharide chains of different molecular weights, which is relatively complicated and is not suitable for the above-mentioned conventional monosaccharide content determination method.
  • gel size exclusion chromatography has many influencing factors, such as poor column durability and relatively high peak spread.
  • the baseline is not easy to balance, and the reproducibility is poor.
  • a new content determination method needs to be established. There is no report on the determination of fucosylated glycosaminoglycans by volumetric method in the existing reports.
  • the purpose of the present invention is to provide a method for measuring the content of low molecular weight fucosylated glycosaminoglycan, which is used to monitor the quality of a low molecular weight fucosylated glycosaminoglycan and ensure the product of low molecular weight fucosylated glycosaminoglycan.
  • the stability and controllability of quality promote the application of low molecular weight fucosylated glycosaminoglycans in medicine, food and other fields.
  • the present invention provides a method for determining the content of low molecular weight fucosylated glycosaminoglycan, wherein the fucosylated glycosaminoglycan has the following structure:
  • R 1 , R 2 , R 3 , R 4 , R 5 are optionally independently -H or -SO 3 H;
  • R 6 is optionally -H, substituted or unsubstituted C1-C6 hydrocarbyl or C7-C12 aryl;
  • R 7 is optionally -H, -SO 3 H, C2-C5 acyl
  • R 8 is optionally a group represented by formula (II), formula (III) or formula (IV):
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 are as defined above;
  • R 9 and R 10 are optionally -H, substituted or unsubstituted C1-C6 hydrocarbyl or C7 - C12 aryl;
  • R 11 is optionally -NHR 12 , -OR 13 , wherein R 12 and R 13 are optionally -H, substituted or unsubstituted C 1 -C 6 hydrocarbyl, or C 7 -C 12 aryl; and n is optionally is a natural number from 0 or 1 to 8;
  • the measurement method is to decationize the low-molecular-weight fucosylated glycosaminoglycan reference substance, add it to titration, record the change of the conductance value during the titration, and measure the volume of the titrant consumed by the reference substance; On the ordinate, draw the titration curve with the volume of the titrant as the abscissa; draw the first, second, and third best straight lines for the three linear parts of the sudden drop, slight climb, and sharp rise of the titration curve respectively; The intersection of the second straight line and the third straight line is a vertical line to the abscissa, and the intersection of the vertical line and the abscissa is the volume of the titrant consumed by sulfate and carboxylate.
  • the titration solution is an alkaline solution, preferably a hydroxide solution.
  • a sodium solution more preferably a sodium hydroxide solution with a concentration of 0.02-0.1 mol/L.
  • V is the volume of the titrant that sulfate and carboxylate consume together in the test sample
  • R is the ratio of the volume of the titrant and the quality of the reference substance consumed by sulfate radicals and carboxylate radicals in the fucosylated glycosaminoglycan reference substance;
  • m is the weight of the fucosylated glycosaminoglycan to be tested, calculated as a dry product.
  • the method for rapidly measuring a low molecular weight fucosified glycosaminoglycan provided by the invention has the advantages of simple operation, good accuracy and economical efficiency. It is very suitable for the promotion and application of product batch inspection and quality control in industrial scale production.
  • Embodiment 1 LFG content determination method verification
  • Table 1 Main equipment information table
  • Table 2 Main reagent information table
  • the content of the reference substance is expressed in terms of HPLC purity (differential detector, area normalized), calculated in terms of dry product, and the titration coefficient is obtained after calibration.
  • the low molecular weight fucosylated glycosaminoglycan test substance used in the examples was extracted from sea cucumber according to the method disclosed in Chinese Patent No. 201410007855.
  • Reference substance solution Weigh an appropriate amount of LFG reference substance, accurately weigh, dissolve in water and dilute to make a solution containing about 5mg per 1ml, shake well, prepare 2 copies in parallel, and set aside.
  • Test solution Weigh an appropriate amount of LFG test product, accurately weigh it, dissolve in water and dilute to make a solution containing about 5mg per 1ml, shake well, prepare 6 copies in parallel, and set aside.
  • the coefficient described in Table 4 is the volume V1 of the sodium hydroxide titration solution consumed by the LFG reference substance, and the amount of sodium hydroxide titration solution consumed per unit volume is calculated, which is equivalent to a certain quality of LFG (calculated as dry product).
  • the average value of the content determination results of this method is 104.16%, and the repeatability RSD value is 2.90%.
  • the method is used as a method for the determination of multi-component samples of fucosylated glycosaminoglycans, and the accuracy of the determination results is good.
  • An example of a titration curve is shown in Figure 1.
  • Embodiment 2 Determination of test product
  • the reagents, consumables and instruments required by the method are all conventional laboratory equipment, and the measurement cost is lower than that of the prior art HPLC measurement method, and the operation is simpler.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)

Abstract

L'invention divulgue un procédé volumétrique de détermination de la teneur en glycosaminoglycane fucosylé de faible poids moléculaire. Un échantillon de référence et un échantillon de test sont soumis séparément à un traitement d'échange cationique, et un agent de titrage alcalin est ajouté à des fins de titrage. Le volume de l'agent de titrage consommé collectivement par le sulfate et le carboxylate est enregistré. La teneur en glycosaminoglycane fucosylé de l'échantillon de test de glycosaminoglycane fucosylé de faible poids moléculaire est obtenue au moyen d'une comparaison et d'un calcul. L'opération est simple, économique et efficace, ce qui rend le procédé approprié pour une production de processus à grande échelle.
PCT/CN2020/117364 2020-09-24 2020-09-24 Procédé de détermination de la teneur en glycosaminoglycane fucosylé de faible poids moléculaire WO2022061637A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115286725A (zh) * 2022-08-31 2022-11-04 山东万邦赛诺康生化制药股份有限公司 一种高纯度低分子量肝素的制备方法

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010030608A1 (fr) * 2008-09-09 2010-03-18 The Government Of The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Procédés de détection et de quantification de glycosaminoglycanes sur-sulfatés
CN102558389A (zh) * 2011-12-22 2012-07-11 中国科学院昆明植物研究所 低分子量岩藻糖化糖胺聚糖的羧基还原衍生物及其制备方法与用途
CN103788222A (zh) * 2014-01-08 2014-05-14 中国科学院昆明植物研究所 Fuc3S4S取代的低聚糖胺聚糖及其制备方法
CN106349407A (zh) * 2016-08-29 2017-01-25 中国海洋大学 一种低分子量岩藻糖化硫酸软骨素及其制备方法和在制备抗特鲁索综合征的药物中的应用
CN108285498A (zh) * 2017-01-10 2018-07-17 九芝堂股份有限公司 一种抑制内源性凝血因子x酶复合物的寡糖化合物及其制备方法与用途

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010030608A1 (fr) * 2008-09-09 2010-03-18 The Government Of The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Procédés de détection et de quantification de glycosaminoglycanes sur-sulfatés
CN102558389A (zh) * 2011-12-22 2012-07-11 中国科学院昆明植物研究所 低分子量岩藻糖化糖胺聚糖的羧基还原衍生物及其制备方法与用途
CN103788222A (zh) * 2014-01-08 2014-05-14 中国科学院昆明植物研究所 Fuc3S4S取代的低聚糖胺聚糖及其制备方法
CN106349407A (zh) * 2016-08-29 2017-01-25 中国海洋大学 一种低分子量岩藻糖化硫酸软骨素及其制备方法和在制备抗特鲁索综合征的药物中的应用
CN108285498A (zh) * 2017-01-10 2018-07-17 九芝堂股份有限公司 一种抑制内源性凝血因子x酶复合物的寡糖化合物及其制备方法与用途

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
"Potentiometric Methods for Determining Chondroitin Sulfate Content", 31 October 1994, ZHEJIANG UNIVERSITY PRESS, CN, ISBN: 7-308-01272-7, article ZHANG, WEIJIE: "Determination of the ratio of sulfuric acid group/mosaic acid group in glycosaminoglycans", pages: 567 - 570, XP009536043 *
LUO, LAN: "Comparsion of Physicochemical Characteristics and Anticoagulant Activities of Sulfated Polysaccharides from Four Species of Sea Cucumbers", CHINESE MASTER’S THESES FULL-TEXT DATABASE, MEDICINE & HEALTH SCIENCES, 1 May 2013 (2013-05-01), XP055914564, [retrieved on 20220421] *
ZHANG, GUANSHUN: "电位法测定硫酸软骨素的含量", STRAIT PHARMACEUTICAL JOURNAL, 30 October 2004 (2004-10-30), XP055914575, [retrieved on 20220421] *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115286725A (zh) * 2022-08-31 2022-11-04 山东万邦赛诺康生化制药股份有限公司 一种高纯度低分子量肝素的制备方法

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