WO2020107820A1 - 一种关节腔注射制剂及其应用 - Google Patents

一种关节腔注射制剂及其应用 Download PDF

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WO2020107820A1
WO2020107820A1 PCT/CN2019/087849 CN2019087849W WO2020107820A1 WO 2020107820 A1 WO2020107820 A1 WO 2020107820A1 CN 2019087849 W CN2019087849 W CN 2019087849W WO 2020107820 A1 WO2020107820 A1 WO 2020107820A1
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xanthan gum
preparation
million
joint cavity
deacetylated
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PCT/CN2019/087849
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English (en)
French (fr)
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凌沛学
刘飞
邵华荣
陈磊
张治云
张晓元
陈启鑫
韩冠英
程艳玲
张建强
张岱州
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山东省药学科学院
山东福瑞达医药集团有限公司
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Priority to US17/297,467 priority Critical patent/US11376274B2/en
Publication of WO2020107820A1 publication Critical patent/WO2020107820A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/716—Glucans
    • A61K31/723—Xanthans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7016—Disaccharides, e.g. lactose, lactulose
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/716—Glucans
    • A61K31/722—Chitin, chitosan
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
    • A61K31/728—Hyaluronic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/737—Sulfated polysaccharides, e.g. chondroitin sulfate, dermatan sulfate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
    • C08B37/0033—Xanthan, i.e. D-glucose, D-mannose and D-glucuronic acid units, saubstituted with acetate and pyruvate, with a main chain of (beta-1,4)-D-glucose units; Derivatives thereof

Definitions

  • the invention relates to the technical field of orthopaedic medicine, in particular to a joint cavity injection preparation and its application.
  • Osteoarthritis is a common degenerative joint disease characterized by the destruction of articular cartilage and bone hyperplasia, and the incidence is higher in the elderly. With the aging population in my country and the increase in the number of obese people, the incidence of OA continues to rise. In OA lesions, the viscoelasticity of joint synovial fluid is significantly reduced, and cartilage is degraded and destroyed, resulting in joint dysfunction.
  • Viscoelastic supplementation is one of the main methods of clinical treatment of OA.
  • HA hyaluronic acid
  • the lubrication and buffering function of joint synovial fluid can be restored, and cartilage tissue repair can be promoted.
  • HA is easily degraded by hyaluronidase and free radicals in the body, and stays in the joint cavity for a short time, therefore, the development of new viscoelastic supplements with similar viscoelasticity and better stability for OA treatment can reduce drug administration It is of great significance to avoid infections caused by repeated injections and improve patient compliance.
  • Xanthan gum is a biopolymer polysaccharide, similar to HA, with better viscoelasticity and better stability.
  • XG is a water-soluble extracellular polysaccharide produced by fermenting Xanthomonas camperstris with black sugar as the main raw material.
  • the relative molecular mass (Mr) of XG is 2 ⁇ 10 6-2 ⁇ 10 7.
  • the basic structure of the XG molecule is composed of pentose repeating units, the main chain is D-glucose connected by ⁇ -1,4 bonds, and the side chain is a trisaccharide formed by alternately connecting D-mannose and D-glucuronic acid.
  • the mannose connected to the main chain is partially acetylated at the 6-position C, and the mannose at the end of some side chains is connected to a pyruvate group at the 6-position C;
  • the secondary structure is that the trisaccharide side chain is wound around the main chain in reverse, through hydrogen The bond maintains a double helix or multi-helix structure;
  • the tertiary structure is a secondary helix structure composed of a network structure through non-covalent bonds.
  • the structure and conformation of xanthan gum determine the functional characteristics of its solution: the complex aggregation structure and intermolecular force of xanthan gum determine its high viscosity at low shear and low concentration, which is more than other polysaccharide solutions.
  • the removal of acetyl groups and the removal of pyruvate groups of xanthan gum will significantly change the properties of xanthan gum.
  • the intermolecular force of xanthan gum after the pyruvate group is removed is significantly reduced, and the pyruvate group may form hydrogen bonds with each other in the xanthan gum molecule and interact with the acetyl of the adjacent side chain.
  • the base generates hydrogen bonds to stabilize the molecular structure of xanthan gum.
  • the acetyl group is usually considered to provide the intramolecular interaction force, so the xanthan gum molecule becomes more compliant after the acetyl group is removed.
  • the XG solution has specific rheological properties, especially the characteristics and viscoelasticity of non-Newtonian fluids: when the joint is at a low impact frequency, it is a viscous solution that acts on the synovial tissue, various tissue planes, ligaments and collagen structures in the joint. Lubrication function reduces friction between tissues. When the joint is at a high impact frequency, the viscous feature is converted to an elastic feature to cushion the impact of stress on the joint.
  • XG can form a macromolecular network in joint synovial fluid, regulate the diffusion of water and macromolecular substances, act as a diffusion barrier in the joint, and act as a barrier to bacteria, toxins, immune complexes, etc., protecting cartilage and synovium Protected from enzymes, chemicals and toxins, it stabilizes cell membranes and inactivates the sensitivity of cell membrane receptors.
  • xanthan gum preparations can perform long-term treatment of OA.
  • Patent application 2017100752628 discloses a pharmaceutical preparation containing low molecular weight xanthan gum for joint cavity injection and a preparation method thereof, and mentions that a pharmaceutical preparation containing a low molecular weight xanthan gum with a relative molecular weight of 100,000 to 1.99 million can be used to prevent rheumatism and Rheumatoid arthritis, osteoarthritis, the protection of articular cartilage in osteoarthritis and the repair of damaged articular cartilage, mainly solve the conventional molecular weight of xanthan gum (relative molecular weight of 2-20 million) due to stability is not easy to be enzymes in the body And the problem of free radical degradation and difficult to be eliminated by the body's metabolism.
  • the low molecular weight xanthan gum drug preparation is used in the treatment of osteoarthritis. Due to the stable structure, it enters the visceral tissue through the blood circulation after joint cavity injection. Dose injection is easy to cause the disadvantages of increased spleen weight and monocyte number, poor biocompatibility, which seriously affects its use effect and hinders further use; while using deacetylated xanthan gum of equivalent molecular weight, even the molecular weight and concentration are more The high deacetylated xanthan gum can solve the above problems while ensuring excellent therapeutic effect.
  • the invention provides a joint cavity injection preparation and its application. It uses deacetylated xanthan gum for the treatment of osteoarthritis. It has excellent therapeutic effect and eliminates the adverse reactions caused by the long-term repeated injection of high-dose xanthan gum. It has excellent biocompatibility, a higher safe dosage range, and safer use, which solves the problems in the existing technology.
  • the present invention provides a joint cavity injection preparation whose active ingredient is deacetylated xanthan gum.
  • the mass percentage of the deacetylated xanthan gum to the volume of the joint cavity injection preparation is 0.01%-10% (w/v), preferably 0.5%-8% (w/v), more preferably 1%-5% (w /v).
  • the molecular weight of the deacetylated xanthan gum is 100,000 to 20 million, preferably 500,000 to 10 million, and more preferably 800 to 3 million.
  • the preparation also includes one or more of sodium hyaluronate, chondroitin sulfate, chitosan or trehalose.
  • the preparation also includes disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and water for injection.
  • the formulation includes the following components by mass volume fraction: deacetylated xanthan gum 1%-5%, disodium hydrogen phosphate 0.4%-0.7%, sodium dihydrogen phosphate 0.1%-0.16%, sodium chloride 0.25%- 0.41%, the balance of water for injection.
  • the pH of the preparation is 5.5-9, and the osmotic pressure is 200-400 mOsmol/L.
  • the present invention also provides the use of deacetylated xanthan gum in the preparation of a medicament for treating osteoarthritis.
  • the biological safety of the medicine for treating osteoarthritis prepared by deacetylated xanthan gum is higher than that of the medicine for treating osteoarthritis prepared by xanthan gum.
  • the biological safety of drugs prepared with deacetylated xanthan gum is superior to xanthan gum.
  • the biosafety of the former is significantly better than that of the latter.
  • the deacetylated xanthan gum and xanthan gum when the dosage is the same (5%), the biological safety of deacetylated xanthan gum is significantly higher than that of xanthan gum.
  • the dosage of the deacetylated xanthan gum in the medicine is 0.01%-10% (w/v), preferably 0.5%-8% (w/v), more preferably 1%-5% (w/v);
  • the molecular weight of the deacetylated xanthan gum is 100,000 to 20 million, preferably 500,000 to 10 million, and more preferably 800 to 3 million.
  • the dosage form of the medicine is selected from injection preparations, gels, ointments, aerosols or sprays.
  • the invention also provides a method for preparing a joint cavity injection preparation, comprising the step of using deacetylated xanthan gum as the active ingredient of the injection preparation.
  • the deacetylated xanthan gum in the present invention may be commercially available deacetylated xanthan gum, or may be prepared by the following preparation method:
  • the preparation step of the deacetylated xanthan gum also includes the step of drying the precipitate of step (3).
  • the pH of the joint cavity preparation prepared by the above method is 5.5-9, and the osmotic pressure is 200-400 mOsmol/L.
  • the present invention uses deacetylated xanthan gum with different conformation and physicochemical properties from xanthan gum instead of xanthan gum, and is used for the production of drugs and preparations for osteoarthritis. Better biocompatibility and safety. Compared with xanthan gum, deacetylated xanthan gum is used in the production of drugs and preparations for osteoarthritis when the molecular weight or molecular weight and concentration are higher. It can also be a good solution for the slow elimination of xanthan gum in the body The resulting problem of poor biocompatibility provides a new way for the development of new drugs such as osteoarthritis drugs and joint cavity injection preparations.
  • the Mw of the deacetylated xanthan gum was detected by multi-angle laser to be 1 million.
  • the hydroxylamine hydrochloride colorimetric method showed that the structure did not contain acetyl groups.
  • the Mw of the deacetylated xanthan gum was detected by multi-angle laser to be 2 million.
  • the hydroxylamine hydrochloride colorimetric method showed that the structure did not contain acetyl groups.
  • the Mw of the deacetylated xanthan gum was detected by multi-angle laser to be 4 million.
  • the hydroxylamine hydrochloride colorimetric method showed that the structure did not contain acetyl groups.
  • the Mw of the deacetylated xanthan gum was detected by multi-angle laser to be 5 million.
  • the hydroxylamine hydrochloride colorimetric method showed that the structure did not contain acetyl groups.
  • the Mw of the deacetylated xanthan gum detected by multi-angle laser was 2.6 million.
  • the hydroxylamine hydrochloride colorimetric method showed that the structure did not contain acetyl groups.
  • deacetylated xanthan gum prepared by the above method is used for the preparation of joint cavity injections:
  • a total of 64 New Zealand white rabbits were randomly divided into 8 groups, 8 in each group, half male and female: normal control group, negative control group (physiological saline), and 1 group of deacetylated xanthan gum (Example 6, 1%) , 1 million), deacetylated xanthan gum 2 groups (Example 7, 3%, 1 million), deacetylated xanthan gum 3 groups (Example 8,5%, 1 million), deacetylated xanthan gum 4 groups (Example 9, 1%, 5 million), ordinary xanthan gum group (1%, 1 million), sodium hyaluronate control group.
  • the knee osteoarthritis model was established by cutting the left knee ligament through anterior cruciate ligament excision. After successful modeling, the corresponding drugs or saline were injected into the joint cavity respectively, and the dosage was 0.3 mL/joint.
  • the sodium hyaluronate control group was administered once a week for a total of 5 times.
  • the deacetylated xanthan gum group and the common xanthan gum group were administered once every 5 weeks for a total of 2 times. Animals in each group They were sacrificed 10 weeks after the first administration. Pathological observation of articular cartilage was performed and scored. The score results are shown in Table 1.
  • a special bovine bone steel knife was used to strip the cartilage of the articular cartilage to make a cartilage block with a uniform thickness (1.89mm).
  • the reciprocating friction tester was used to detect the tribological index under the lubrication of different injections, including the dry friction group, saline group, and glass acid.
  • deacetylated xanthan gum 1 group (Example 6, 1%, 1 million), deacetylated xanthan gum 2 groups (Example 7, 3%, 1 million), deacetylated xanthan gum 3 groups (implemented Example 8, 5%, 1 million), deacetylated xanthan gum 4 groups (Example 9, 1%, 5 million), ordinary xanthan gum group (1%, 1 million), sodium hyaluronate control group.
  • Test machine setting parameters test force is 3N, reciprocating frequency is 1Hz. The test results are shown in Table 2 below.
  • Eighty SD rats were randomly divided into 8 groups, 10 in each group, half male and female: negative control group (physiological saline), deacetylated xanthan gum group 1 (Example 6, 1%, 1 million), deacetylated Xanthan gum 2 groups (Example 7, 3%, 1 million), deacetylated xanthan gum 3 groups (Example 8, 5%, 1 million), deacetylated xanthan gum 4 groups (Example 9, 1% , 5 million), deacetylated xanthan gum 5 groups (Example 10, 5%, 5 million), ordinary xanthan gum 1 group (5%, 1 million), ordinary xanthan gum 2 groups (5%, 5 million) ), and the corresponding drugs or physiological saline were injected into the joint cavity respectively, the dosage was 0.06 mL/joint, and the drug was administered once every 2 weeks for 6 consecutive months. Two weeks after the last administration, the anesthetized animal was bled for routine blood examination. After

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Abstract

一种关节腔注射制剂及其应用。该关节腔注射制剂采用脱乙酰黄原胶，与现有黄原胶关节腔注射制剂相比，具有更好的生物相容性，安全剂量范围更高。

Description

一种关节腔注射制剂及其应用 技术领域
本发明涉及骨科医药技术领域，尤其是涉及一种关节腔注射制剂及其应用。
背景技术
骨关节炎(osteoarthritis，OA)是一种以关节软骨破坏及骨质增生为特征的常见退行性关节疾病，老年人发病率较高。随着我国人口老龄化及肥胖人口数量的增加，OA发病率持续上升。OA病变时，关节滑液的黏弹性显著降低，软骨发生降解破坏，从而导致关节出现功能障碍。
黏弹性补充疗法(viscosupplementation)是OA临床治疗主要方法之一，通过向关节腔内注射外源性透明质酸(hyaluronic acid，HA)，可恢复关节滑液的润滑缓冲功能，促进软骨组织修复，缓解OA症状，改善关节的生理功能。由于HA易被体内的透明质酸酶及自由基降解，在关节腔内存留时间较短，因此，开发黏弹性相似、稳定性更好的新型黏弹性补充剂用于OA治疗，可减少给药次数，避免反复注射引起的感染，提高患者顺应性，具有重要意义。
黄原胶(xanthan gum,XG)是一种生物高分子多糖，与HA相似，具有较好的黏弹性，稳定性更好。XG是甘蓝黑腐病黄单胞菌(Xanthomonascamperstris)以糖类为主要原料发酵产生的一种水溶性胞外多糖。XG的相对分子质量(relative molecular mass，Mr)为2×10 6-2×10 7。XG分子的基本结构是由戊糖重复单元构成，主链是β-1,4键连接的D-葡 萄糖，侧链为D-甘露糖与D-葡糖醛酸交替连接而成的三糖，其中连接主链的甘露糖6位C部分乙酰化，部分侧链末端的甘露糖4，6位C连接1个丙酮酸基团；二级结构是三糖侧链反向缠绕主链，通过氢键维系成的双螺旋或多螺旋结构；三级结构是二级螺旋结构通过非共价键组成网状结构。黄原胶的结构和构象决定了其溶液的功能特性：黄原胶复杂的聚集态结构及分子间作用力决定了其溶液在低剪切、低浓度下具有高粘度，相对其它多糖溶液具有更高的模量，以及具有假塑性行为；黄原胶硬直分子链和分子链上具备的氢键、阴离子，侧链缠结起来对主链的保护，使其溶液具备良好的耐热和耐盐性能，并且对酸碱以及酶解也具有良好的稳定性。一般而言，黄原胶中丙酮酸取代基的含量在30-40％之间，乙酰化的基团在60-70％之间；两者在链上的分布并无规律，但是对于黄原胶的构象及物化性质却有着很大的影响。黄原胶脱去乙酰基和脱去丙酮酸基团都会使黄原胶的性质发生显著变化。据流变学研究，脱去丙酮酸基团后的黄原胶分子间作用力显著减小，丙酮酸基团在黄原胶分子中可能相互之间形成氢键，并与邻近侧链的乙酰基产生氢键，以此来稳定黄原胶的分子结构。而乙酰基团通常被认为是提供了分子内的相互作用力，因此脱去乙酰基后黄原胶分子变得更加柔顺。
本实验室前期研究发现，关节腔注射XG对实验性OA具有长效治疗作用，可减少次数，具有开发成为OA治疗新药的潜力。XG溶液具有特异的流变学性质，尤其是非牛顿流体的特性和黏弹性：当关节处于低撞击频率时，呈粘性溶液，对关节内的滑膜组织、各组织平面、韧带和胶原结构等行使润滑功能，减少组织间的摩擦，当关节处于高撞击频率 时，由粘性特征转换为弹性特征，缓冲应力对关节的撞击。而XG在关节滑液中，可形成大分子网络结果，调节水和大分子物质的扩散，在关节内充当一扩散屏障，对细菌、毒素、免疫复合物等起屏障作用，保护软骨和滑膜免受酶、化学物质以及毒素等的破坏，稳定细胞膜、钝化细胞膜感受器的敏感性。现有技术已有黄原胶制剂能对OA进行长效治疗的报道。专利申请2017100752628公开了关节腔注射用的含低分子量黄原胶的药物制剂及其制备方法，提及含有相对分子质量为10万-199万的低分子量黄原胶药物制剂可用于预防风湿、类风湿性关节炎、骨关节炎，保护骨关节炎中关节软骨以及修复受损的关节软骨的作用，主要解决了常规分子量的黄原胶(相对分子量为200-2000万)由于稳定不易被体内酶和自由基降解、难以由机体代谢排出的问题。然而，低分子量黄原胶药物制剂在用于骨关节炎治疗中，由于结构稳定，关节腔注射后通过血液循环进入内脏组织，在组织中存留时间相对较长，清除较慢，且长期反复高剂量注射易引起脾脏重量增加、单核细胞数增加的弊端，生物相容性差，严重影响了其使用效果，阻碍了进一步使用；而采用相当分子量的脱乙酰基黄原胶，甚至分子量及浓度更高的脱乙酰基黄原胶，在保证优异治疗效果的同时，能很好的解决上述问题。
发明内容
本发明提供了一种关节腔注射制剂及其应用，采用脱乙酰基黄原胶用于骨关节炎的治疗，治疗功效优异，消除了现有长期反复注射高剂量黄原胶引起的不良反应，具备优异的生物相容性，安全剂量范围更高，使用更安全，解决了现有技术中存在的问题。
一方面，本发明提供了一种关节腔注射制剂，所述制剂的活性成分为脱乙酰基黄原胶。
所述脱乙酰基黄原胶质量占关节腔注射制剂体积的百分数为0.01％-10％(w/v)，优选0.5％-8％(w/v)，更优选1％-5％(w/v)。
所述脱乙酰基黄原胶的分子量为10万-2000万，优选50-1000万，更优选80-300万。
所述制剂还包括透明质酸钠、硫酸软骨素、壳聚糖或海藻糖的一种或几种。
所述制剂还包括磷酸氢二钠、磷酸二氢钠、氯化钠和注射用水。
所述制剂包括如下质量体积分数的各成分：脱乙酰基黄原胶1％-5％、磷酸氢二钠0.4％-0.7％、磷酸二氢钠0.1％-0.16％、氯化钠0.25％-0.41％、注射用水余量。
所述制剂的pH为5.5-9，渗透压为200-400mOsmol/L。
另一方面，本发明还提供了脱乙酰基黄原胶在制备治疗骨关节炎的药物中的用途。
脱乙酰基黄原胶制备的治疗骨关节炎的药物的生物安全性高于黄原胶制备得到的治疗骨关节炎的药物的生物安全性。总体来说，无论脱乙酰基黄原胶和普通黄原胶的分子量和添加剂量如何，采用脱乙酰基黄原胶制备得到的药物的生物安全性要优于黄原胶。尤其是，当采用相当剂量和相当分子量的脱乙酰基黄原胶和普通黄原胶制备治疗骨关节炎的药物时，前者的生物安全性要显著优于后者，例如，采用平均分子量为100万的脱乙酰基黄原胶和黄原胶，当添加剂量相同(5％)时，脱乙酰基黄原 胶的生物安全性要显著高于黄原胶。
所述脱乙酰基黄原胶在药物中用量为0.01％-10％(w/v)，优选0.5％-8％(w/v)，更优选1％-5％(w/v)；所述脱乙酰基黄原胶的分子量为10万-2000万，优选50-1000万，更优选80-300万。
所述药物的剂型选自注射制剂、凝胶剂、软膏剂、气雾剂或喷雾剂。
本发明还提供了一种制备关节腔注射制剂的方法，包含以脱乙酰基黄原胶作为所述注射制剂的活性成分的步骤。本发明中的脱乙酰基黄原胶可以采用市售的脱乙酰基黄原胶，也可以采用以下的制备方法制备得到：
(1)将质量分数1％-10％、相对分子量为100-2000万的黄原胶水溶液调节pH至10-13，在温度50-100℃条件下搅拌0.2-2h；
(2)调节pH至1.5-7，置于0.05-0.15MPa下反应10-30min；
(3)冷却后调节pH至中性，使用乙醇沉淀，取沉淀物，即得。
上述脱乙酰基黄原胶的制备步骤还包括对步骤(3)的沉淀物进行干燥的步骤。
上述方法制得的关节腔注射制剂的pH为5.5-9，渗透压为200-400mOsmol/L。
本发明的有益效果：
本发明通过采用构象及物化性质与黄原胶不同的脱乙酰基黄原胶替代黄原胶，用于骨关节炎用药物及制剂的生产，所得治疗骨关节炎的药物及关节腔注射制剂具备更好的生物相容性和安全性。脱乙酰基黄原胶相比黄原胶，相当分子量或分子量及浓度更高时，用于骨关节炎用药物 及制剂的生产，也能很好的解决因黄原胶结构稳定在体内清除慢造成的生物相容性差的问题，为骨关节炎用药物及关节腔注射制剂等新药的开发提供了新途径。
具体实施方式
为为能清楚说明本方案的技术特点，下面通过具体实施方式，对本发明进行详细阐述。
实施例1
脱乙酰基黄原胶的制备方法：
取10g分子量为300万的黄原胶充分溶解于1000mL水中，调节pH 11，在温度50℃下搅拌0.5h后，调节pH 2.0，置于0.1MPa下反应20min，冷却后调节pH至7.0，使用乙醇沉淀，沉淀产物采用40℃减压干燥24h，即得。
经多角度激光检测该脱乙酰基黄原胶的Mw为100万，采用盐酸羟胺比色法检测显示其结构中不含乙酰基。
实施例2
脱乙酰基黄原胶的制备方法：
取20g分子量为300万的黄原胶充分溶解于1000mL水中，调节pH 11，在温度60℃下搅拌2h后，调节pH 3.0，置于0.1MPa下反应30min，冷却后调节pH至7.0，使用乙醇沉淀，沉淀产物采用40℃减压干燥24h，即得。
经多角度激光检测该脱乙酰基黄原胶的Mw为200万，采用盐酸羟胺比色法检测显示其结构中不含乙酰基。
实施例3
脱乙酰基黄原胶的制备方法：
取30g分子量为500万的黄原胶充分溶解于1000mL水中，调节pH 12， 在温度50℃下搅拌1h后，调节pH 5.0，置于0.1MPa下反应20min，冷却后调节pH至7.0，使用乙醇沉淀，沉淀产物采用40℃减压干燥24h，即得。
经多角度激光检测该脱乙酰基黄原胶的Mw为400万，采用盐酸羟胺比色法检测显示其结构中不含乙酰基。
实施例4
脱乙酰基黄原胶的制备方法：
取50g分子量为500万的黄原胶充分溶解于1000mL水中，调节pH 11，在温度55℃下搅拌0.5h后，调节pH 7.0，置于0.1MPa下反应20min，冷却后调节pH至7.0，使用乙醇沉淀，沉淀产物采用40℃减压干燥24h，即得。
经多角度激光检测该脱乙酰基黄原胶的Mw为500万，采用盐酸羟胺比色法检测显示其结构中不含乙酰基。
实施例5
脱乙酰基黄原胶的制备方法：
取50g分子量为800万的黄原胶充分溶解于1000mL水中，调节pH 11，在温度55℃下搅拌0.5h后，调节pH 2.0，置于0.1MPa下反应30min，冷却后调节pH至7.0，使用乙醇沉淀，沉淀产物采用40℃减压干燥24h，即得。
经多角度激光检测该脱乙酰基黄原胶的Mw为260万，采用盐酸羟胺比色法检测显示其结构中不含乙酰基。
将上述方法制得的脱乙酰基黄原胶用于关节腔注射剂的制备：
实施例6
1％脱乙酰基黄原胶关节腔注射制剂
Figure PCTCN2019087849-appb-000001
Figure PCTCN2019087849-appb-000002
实施例7
3％脱乙酰基黄原胶关节腔注射制剂
Figure PCTCN2019087849-appb-000003
实施例8
5％脱乙酰基黄原胶关节腔注射制剂
Figure PCTCN2019087849-appb-000004
实施例9
1％脱乙酰基黄原胶关节腔注射制剂
Figure PCTCN2019087849-appb-000005
Figure PCTCN2019087849-appb-000006
实施例10
5％脱乙酰基黄原胶关节腔注射制剂
Figure PCTCN2019087849-appb-000007
实验研究：
一、对上述脱乙酰基黄原胶关节腔注射制剂进行骨关节炎治疗效果研究的动物试验：
取新西兰大耳白兔64只，随机分为8组，每组8只，雌雄各半：正常对照组，阴性对照组(生理盐水)、脱乙酰黄原胶1组(实施例6，1％，100万)、脱乙酰黄原胶2组(实施例7，3％，100万)、脱乙酰黄原胶3组(实施例8,5％，100万)、脱乙酰黄原胶4组(实施例9，1％，500万)、普通黄原胶组(1％，100万)、玻璃酸钠对照组。
通过前交叉韧带切断术切断左膝韧带建立膝骨关节炎模型，造模成功后，分别关节腔注射给予相应的药物或生理盐水，给药量为0.3mL/关节。其中玻璃酸钠对照组，每周给药1次，共给药5次，脱乙酰黄原胶组及普通黄原胶组，每5周给药1次，共给药2次，各组动物分别于第1次给药10周后处死，进行关节软骨病理学观察，进行评分，评分结果见表1。结果表明，每5周给药1次，脱乙酰基黄原胶与普通黄原胶对于骨关节炎均具有明显的治疗效果，与玻璃酸钠组每周给药1次比较，未见显著性差异。关节软骨病理评分如下表1。
表1 关节软骨病理评分
Figure PCTCN2019087849-appb-000008
二、对上述脱乙酰基黄原胶关节腔注射制剂进行牛关节软骨润滑效果的影响的研究：
采用特殊牛骨钢刀剥离牛关节软骨，制成厚度均匀(1.89mm)的软骨块，于往复摩擦试验机分别检测在不同注射剂润滑下摩擦学指标，包括干摩擦组、生理盐水组、玻璃酸钠组、脱乙酰黄原胶1组(实施例6，1％，100万)、脱乙酰黄原胶2组(实施例7，3％，100万)、脱乙酰黄原胶3组(实施例8，5％，100万)、脱乙酰黄原胶4组(实施例9,1％，500万)、普通黄原胶组(1％，100万)、玻璃酸钠对照组。试验机设置参数：试验力为3N、往复频率为1Hz。测试结果如下表2。
表2 对牛关节软骨润滑效果的影响
Figure PCTCN2019087849-appb-000009
由结果可知，在往复摩擦试验机上，牛膝关节软骨在脱乙酰基黄原胶及普通黄原胶润滑下的润滑效果均优于玻璃酸钠。
三、对上述脱乙酰基黄原胶关节腔注射制剂进行生物相容性研究：
取SD大鼠80只随机分为8组，每组10只，雌雄各半：阴性对照组(生理盐水)、脱乙酰黄原胶1组(实施例6，1％，100万)、脱乙酰黄原胶2组(实施例7，3％，100万)、脱乙酰黄原胶3组(实施例8，5％，100万)、脱乙酰黄原胶4组(实施例9,1％，500万)、脱乙酰黄原胶5组(实施例10，5％，500万)、普通黄原胶1组(5％，100万)、普通黄原胶2组(5％，500万)，分别关节腔注射给予相应的药物或生理盐水，给药量为0.06mL/关节，每2周给药1次，连续给药6个月。末次给药后2周，麻醉动物取血进行血常规检查，处死后，取动物脾脏称重，计算脏器系数(％)。
结果显示，注射普通黄原胶组可引起脾脏重量及单核细胞比率的显著增加，而反复注射高剂量脱乙酰基黄原胶未见脏器组织的不良反应及血常规检查的异常。
以上试验表明，长期反复注射高剂量黄原胶可引起脾脏重量增加和单核细胞增加的不良反应，而长期反复注射高剂量脱乙酰基黄原胶未见不良反应，这表明脱乙酰基黄原胶用于OA治疗的安全剂量范围明显高于普通黄原胶，具有显著的生物相容性优势。
表3 脱乙酰基黄原胶对单核细胞的影响
Figure PCTCN2019087849-appb-000010
Figure PCTCN2019087849-appb-000011
表4 脱乙酰基黄原胶对脾脏重量的影响
Figure PCTCN2019087849-appb-000012
上述具体实施方式不能作为对本发明保护范围的限制，对于本技术领域的技术人员来说，对本发明实施方式所做出的任何替代改进或变换均落在本发明的保护范围内。
本发明未详述之处，均为本技术领域技术人员的公知技术。

Claims (10)

  1. 一种关节腔注射制剂，其特征在于，所述制剂的活性成分为脱乙酰基黄原胶。
  2. 根据权利要求1所述的一种关节腔注射制剂，其特征在于，所述脱乙酰基黄原胶质量占关节腔注射制剂体积的百分数为0.01％-10％(w/v)，优选0.5％-8％(w/v)，更优选1％-5％(w/v)。
  3. 根据权利要求1所述的一种关节腔注射制剂，其特征在于，所述脱乙酰基黄原胶的分子量为10万-2000万，优选50-1000万，更优选80-300万。
  4. 根据权利要求1所述的一种关节腔注射制剂，其特征在于，所述制剂还包括透明质酸钠、硫酸软骨素、壳聚糖或海藻糖的一种或几种。
  5. 根据权利要求1所述的一种关节腔注射制剂，其特征在于，所述制剂的pH为5.5-9，渗透压为200-400mOsmol/L。
  6. 脱乙酰基黄原胶在制备治疗骨关节炎的药物中的用途。
  7. 根据权利要求6所述的脱乙酰基黄原胶在制备治疗骨关节炎的药物中的用途，其特征在于，所述药物的生物安全性高于黄原胶制备得到的治疗骨关节炎的药物的生物安全性。
  8. 根据权利要求6所述的脱乙酰基黄原胶在制备治疗骨关节炎的药物中的用途，其特征在于，所述脱乙酰基黄原胶在药物中用量为0.01％-10％(w/v)，优选0.5％-8％(w/v)，更优选1％-5％(w/v)；所述脱乙酰基黄原胶的分子量为10万-2000万，优选50-1000万，更优选80-300万。
  9. 根据权利要求6所述的脱乙酰基黄原胶在制备治疗骨关节炎的药物中的用途，其特征在于，所述药物的剂型选自注射制剂、凝胶剂、软膏剂、气雾剂或喷雾剂。
  10. 一种制备关节腔注射制剂的方法，其特征在于，包含以脱乙酰基黄原胶作为所述注射制剂的活性成分的步骤。
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Publication number Priority date Publication date Assignee Title
CN101940587A (zh) * 2009-07-03 2011-01-12 山东省生物药物研究院 一种含有黄原胶的关节腔内注射用药物制剂
CN102946855A (zh) * 2010-06-23 2013-02-27 爱尔康研究有限公司 含有妥布霉素和地塞米松的局部用眼科混悬剂
CN105251048A (zh) * 2015-10-16 2016-01-20 山东福瑞达医药集团公司 一种含黄原胶的可注射型软骨组织保护及修复材料
CN106692179A (zh) * 2017-02-13 2017-05-24 山东省药学科学院 关节腔注射用含低分子量黄原胶的药物制剂及其制备方法
CN109260219A (zh) * 2018-11-30 2019-01-25 山东省药学科学院 一种关节腔注射制剂及其应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8623839B2 (en) * 2011-06-30 2014-01-07 Depuy Mitek, Llc Compositions and methods for stabilized polysaccharide formulations

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101940587A (zh) * 2009-07-03 2011-01-12 山东省生物药物研究院 一种含有黄原胶的关节腔内注射用药物制剂
CN102946855A (zh) * 2010-06-23 2013-02-27 爱尔康研究有限公司 含有妥布霉素和地塞米松的局部用眼科混悬剂
CN105251048A (zh) * 2015-10-16 2016-01-20 山东福瑞达医药集团公司 一种含黄原胶的可注射型软骨组织保护及修复材料
CN106692179A (zh) * 2017-02-13 2017-05-24 山东省药学科学院 关节腔注射用含低分子量黄原胶的药物制剂及其制备方法
CN109260219A (zh) * 2018-11-30 2019-01-25 山东省药学科学院 一种关节腔注射制剂及其应用

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