WO2008037171A1 - A process for preparing a hyaluronic acid gel in the two crosslinking stages - Google Patents

A process for preparing a hyaluronic acid gel in the two crosslinking stages Download PDF

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Publication number
WO2008037171A1
WO2008037171A1 PCT/CN2007/002466 CN2007002466W WO2008037171A1 WO 2008037171 A1 WO2008037171 A1 WO 2008037171A1 CN 2007002466 W CN2007002466 W CN 2007002466W WO 2008037171 A1 WO2008037171 A1 WO 2008037171A1
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hyaluronic acid
gel
concentration
acid gel
preparing
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PCT/CN2007/002466
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French (fr)
Chinese (zh)
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Min Liu
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Beijing Pro-Medical Biotech Research Institute
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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
    • A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
    • A61K31/728—Hyaluronic acid
    • 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
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
    • C08J3/244—Stepwise homogeneous crosslinking of one polymer with one crosslinking system, e.g. partial curing
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L5/00—Compositions of polysaccharides or of their derivatives not provided for in groups C08L1/00 or C08L3/00
    • C08L5/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
    • C08J2305/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof

Definitions

  • the present invention relates to a hyaluronic acid gel and a process for the preparation thereof, and more particularly to a hyaluronic acid gel having a long retention time and its passage A method of preparing the hyaluronic acid gel by secondary crosslinking.
  • Hyaluronic acid is a linear polymer polysaccharide in which ⁇ -DN acetylglucosamine and ⁇ -D-glucuronic acid are bonded to each other, and is widely distributed in mammalian connective tissues, cockscombs, and streptococcus.
  • hyaluronic acid gel prepared from hyaluronic acid as a filler is transplanted or injected into the body to show good body compatibility, and it has the functions of anti-wrinkle, breast enhancement and filling. , and has no side effects on the human body, and is widely used in medical and beauty industries.
  • This patent is to overcome the shortcomings of the above-mentioned hyaluronic acid gel storage time, and the secondary cross-linking process effectively increases the steric hindrance, so that the prepared hyaluronic acid gel is attacked by hyaluronidase. Difficult to enter, greatly improve the retention time of hyaluronic acid gel.
  • the hyaluronic acid gel of the present invention has a retention time of 2 to 50 times that of the prior art hyaluronic acid gel.
  • One object of the present invention is to provide a hyaluronic acid gel which has a long storage time and a good liner effect and a preparation method thereof, and the storage time of the prepared hyaluronic acid gel is obtained by secondary crosslinking. 2 to 50 times that of a cross-linked hyaluronic acid gel.
  • Another object of the present invention is to provide a medical material of hyaluronic acid gel.
  • a method for preparing a hyaluronic acid gel comprising the steps of:
  • the sodium hyaluronate is dissolved in a sodium hydroxide solution having a concentration of 0.3% to 0.9%, the concentration of hyaluronic acid is 6%-10%, preferably 6%-8%, and the weight of sodium hyaluronate is 0.5-2.8 wt. % glycidyl ether type crosslinking agent, preferably 1-2.5 wt.%, more preferably 1-2 wt.%, reaction temperature 40-60 ° C, preferably 45-55 ° C, reaction time 1-10 hours, placed cool down;
  • the reactant of the step (1) is added to the same weight and the same concentration of the sodium hydroxide solution in the primary crosslinking reaction;
  • the reactant of the step (2) is added to the same or different glycidyl ether type crosslinking agent as (1), and the amount thereof is 3-7 wt.%, preferably 3.75-6.25 t%, more preferably the weight of the sodium hyaluronate. 3-5 wt.% b Stir well to make the solution uniform. Heating is started, the reaction time is 1-5 hours, the reaction temperature is 40-60 ° C, preferably 50-60 ° C, more preferably 50-53 ° C, gel formation;
  • the gel of the step (3) is immersed in a NaCl solution having a concentration of 0.8-1%, and neutralized with a HC1 solution to a pH of 6.5-7.5.
  • the soaking in the step (4) is to soak the gel to a content of hyaluronic acid of 1 to 3 wt%, preferably 1 to 2% w, based on the total weight of the gel.
  • the preparation method further comprises the step of pulverizing when the gel is soaked to a content of hyaluronic acid of 1 to 3 wt.% of the total weight of the gel, more preferably when the gel is soaked to a content of hyaluronic acid as a total gel.
  • a pulverization step is carried out, wherein the glycidyl ether type crosslinking agent is ethylene glycol diglycidyl ether and/or 1,4-butanediol glycidyl ether. .
  • the mass concentration of the NaCl solution described in the step (4) is 0.8-1%; and the mass concentration of the HC1 is 0.3-0.9%.
  • a hyaluronic acid gel prepared by the above preparation method.
  • a medical material comprising a wrinkle pad material, which is formed by immersing and immersing the hyaluronic acid gel after the above-mentioned hyaluronic acid gel.
  • the soaking is immersed in a NaCi solution, and the neutralization is performed by HCL.
  • the solution was neutralized to a pH of 6.5-7.5.
  • the mass concentration of the NaCl solution is 0.8-1%%; and the mass concentration of the HC1 is 0.3-0.9%.
  • the content of hyaluronic acid after soaking the hyaluronic acid gel is 1-3 wt% of the total weight of the gel. / 0 .
  • the hyaluronic acid gel of the present invention is completed by secondary crosslinking, and the amount of the crosslinking agent can be remarkably reduced, and the residue of the crosslinking agent can be reduced, and the residue of the crosslinking agent can be minimized to the injection site.
  • the influence of the human body effectively preventing the damage of the cross-linking agent to the human body.
  • the preservation time of the hyaluronic acid gel is effectively improved, which improves the retention at the injectable site, increases the retention time, and improves the gel quality.
  • the hyaluronic acid gel of the present invention can be used as a cushioning material for wrinkles for use in the medical or cosmetic industry.
  • BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below with reference to examples:
  • Example 1 8 g of sodium hyaluronate was dissolved in 100 g of a sodium hydroxide solution having a concentration of 0.8%, and the concentration of hyaluronic acid was about 8%, and 1.4- 0.08 g of butanediol glycidyl ether was heated at 45 ° C for 3 hours. Place and add the above-mentioned sodium hydroxide solution of the same weight and the same concentration. Then, 0.5 g of 1.4-butylene glycol glycidyl ether was further added, stirred well, and heated at a temperature of 50 ° C for 2.5 hours to prepare a hyaluronic acid gel.
  • the hyaluronic acid gel prepared above was immersed in a 0.9% NaCl solution and neutralized to a pH of 6.5 with a 0.3% HCl solution, and the gel was soaked to a hyaluronic acid content of 1%.
  • the pulverization into the corresponding granules is an ideal crease liner material.
  • the hyaluronic acid gel was subjected to hyaluronidase degradation experiments and compared with primary crosslinked gel particles using the same hyaluronic acid and a crosslinking agent, and as a result, the secondary crosslinked gel particles were degraded. 20.92 times for one cross-linking.
  • Example 2 8 g of sodium hyaluronate was dissolved in 120 g of a sodium hydroxide solution having a concentration of 0.6%, and the concentration of hyaluronic acid was about 6.7%.
  • 0.2 g of ethylene glycol glycidyl ether was added thereto, and the mixture was heated at 50 ° C for 2 hours. Place and add the above-mentioned sodium hydroxide solution of the same weight and the same concentration. Then, 0.3 g of an ethylene glycol glycidyl ether crosslinking agent was further added, and the mixture was thoroughly stirred, and heated at 53 ° C for 1.5 hours to prepare a hyaluronic acid gel.
  • the hyaluronic acid gel prepared above was immersed in a 0.8% NaCl solution, and neutralized to a pH of 7, with a concentration of 0.6% of the HC1 solution, soaked to a hyaluronic acid content of 2%.
  • the pulverization into the corresponding granules is an ideal crease liner material.
  • the hyaluronic acid gel was subjected to hyaluronidase degradation experiments and compared with primary crosslinked gel particles using the same hyaluronic acid and a crosslinking agent, and as a result, the secondary crosslinked gel particles were degraded. 46.65 times as much as one cross-linking.
  • Example 3 8 g of sodium hyaluronate was dissolved in 90 g of a sodium hydroxide solution having a concentration of 0.3%, and the concentration of hyaluronic acid was about 8.9%, and 0.15 g of 1.4-butylene glycol glycidyl ether was added. Heat at 55 ° C for 2 hours and place. Then add the same weight, the same concentration of sodium hydroxide solution, stir well, then add 0.43g ethylene glycol glycidyl ether crosslinker, stir well, react at temperature 55 ° C for 1.5 hours, to produce hyaluronic acid gel.
  • the hyaluronic acid gel prepared above was immersed in a NaCl solution having a concentration of 1%, and neutralized to a pH of 7.5 with a 0.9% HC1 solution, and the gel was soaked to a hyaluronic acid content of 3%.
  • the powder is broken into the corresponding particles, which is the ideal wrinkle pad material.
  • the hyaluronic acid gel was subjected to hyaluronidase degradation experiments and compared with primary crosslinked gel particles using the same hyaluronic acid and a crosslinking agent, and as a result, the secondary crosslinked gel particles were degraded. 10.88 times for one cross-linking.
  • this example only uses ethylene glycol glycidyl ether crosslinker and 1.4-butanediol shrinkage.
  • the specific process of the technical solution of the present invention is described by taking glyceryl ether as an example.
  • any glycidyl ether type crosslinking agent can realize the technical solution according to the art.
  • the preparation method of the hyaluronic acid is universal for any glycidyl ether type crosslinking agent, and the use of any glycidyl ether as a crosslinking agent can obviously obtain the contents of the present invention. Strong support.

Abstract

A process for preparing a hyaluronic acid gel is disclosed,which includes: (1) crosslinking sodium hyaluronate with diglycidyl ether in a NaOH solution, (2) adding the product of (1) to the NaOH solution with the same mass and concentration as the NaOH solution of (1), (3) further crosslinking by adding diglycidyl ether to the product of (2),and thereby forming a gel, (4) soaking the gel of (3) in a NaCl solution and adjusting the solution to pH 6.5-7.5. A hyaluronic acid gel is prepared by the process.A medical materail is made of the hyaluronic acid gel.

Description

两歩交联制备诱明质酸凝胶的¾^ 技术领域 本发明涉及一种透明质酸凝胶及其制备方法， 更具体来说涉及一种保留 时间长的透明质酸凝胶及其通过二次交联制备该透明质酸凝胶的方法。 背景技术 透明质酸是 β-D-N乙酰氨基葡萄糖和 β-D-葡萄糖醛酸相互结合的直链状 高分子多糖， 广泛分布于哺乳动物的结締组织、 鸡冠和链球菌的夹膜等地方， 由于不具有种属及脏器特异性， 由透明质酸制得的透明质酸凝胶作为填充物 移植或注入肌体都显示良好的肌体相容性， 起到抗皱、 丰乳、 充垫等作用， 且对人体无副作用， 广泛应用于医疗、 美容行业。  FIELD OF THE INVENTION The present invention relates to a hyaluronic acid gel and a process for the preparation thereof, and more particularly to a hyaluronic acid gel having a long retention time and its passage A method of preparing the hyaluronic acid gel by secondary crosslinking. BACKGROUND ART Hyaluronic acid is a linear polymer polysaccharide in which β-DN acetylglucosamine and β-D-glucuronic acid are bonded to each other, and is widely distributed in mammalian connective tissues, cockscombs, and streptococcus. Due to the lack of species and organ specificity, hyaluronic acid gel prepared from hyaluronic acid as a filler is transplanted or injected into the body to show good body compatibility, and it has the functions of anti-wrinkle, breast enhancement and filling. , and has no side effects on the human body, and is widely used in medical and beauty industries.
现有技术中与本发明相类似的只有另一专利 CN1590444A,该专利是用透 明质酸 （HA) 与交联剂 （BDDE) 在 15-35°C条件下， 用适当的比例， 反应 2-24小时， 一次交联形成透明质酸凝胶。 其弊端在于， 在专利公开的温度下， 很难形成稳定的凝胶， 所形成的凝胶容易受到透明质酸酶的攻击， 保存时间 短。 而现在的美容领域正日新月异的发展， 因此对既安全又能保存很长时间 的打针除皱原料的需求很迫切。 本专利就是为了克服上述透明质酸凝胶保存 时间短的弊端， 釆用二次交联的工艺有效地增加空间位阻， 使制备的透明质 酸凝胶在受到透明质酸酶攻击时， 很难进入， 极大地提高透明质酸凝胶的保 留时间。 经中国农业大学实验证明， 本发明的透明质酸凝胶的保留时间是现 有技术中透明质酸凝胶的 2— 50倍。 发明内容 本发明的目的之一是提供一种保存时间长、 衬垫效果好的透明质酸凝胶 及其制备方法， 通过二次交联， 使得所制备的透明质酸凝胶的保存时间为一 次交联的透明质酸凝胶的 2— 50倍。 本发明的另一目的是提供一种透明质酸凝胶的医用材料。 In the prior art, similar to the present invention, there is only another patent CN1590444A, which uses hyaluronic acid (HA) and a crosslinking agent (BDDE) at a reaction ratio of 15-35 ° C in a suitable ratio 2- 24 hours, one cross-linking to form a hyaluronic acid gel. The disadvantage is that at the temperature disclosed in the patent, it is difficult to form a stable gel, and the gel formed is easily attacked by hyaluronidase, and the storage time is short. Nowadays, the beauty field is developing with each passing day. Therefore, the demand for injection and wrinkle-removing materials that are safe and can be preserved for a long time is urgent. This patent is to overcome the shortcomings of the above-mentioned hyaluronic acid gel storage time, and the secondary cross-linking process effectively increases the steric hindrance, so that the prepared hyaluronic acid gel is attacked by hyaluronidase. Difficult to enter, greatly improve the retention time of hyaluronic acid gel. According to experiments conducted by the Agricultural University of China, the hyaluronic acid gel of the present invention has a retention time of 2 to 50 times that of the prior art hyaluronic acid gel. SUMMARY OF THE INVENTION One object of the present invention is to provide a hyaluronic acid gel which has a long storage time and a good liner effect and a preparation method thereof, and the storage time of the prepared hyaluronic acid gel is obtained by secondary crosslinking. 2 to 50 times that of a cross-linked hyaluronic acid gel. Another object of the present invention is to provide a medical material of hyaluronic acid gel.
本发明的目的是这样实现的：  The object of the invention is achieved in this way:
一种透明质酸凝胶的制备方法， 包括以下步骤：  A method for preparing a hyaluronic acid gel, comprising the steps of:
( 1 ) 一次交联  (1) One cross-linking
将透明质酸钠溶于浓度 0.3%— 0.9%的氢氧化钠溶液中， 透明质酸的浓 度在 6%— 10%, 优选 6%-8%，加入透明质酸钠重量的 0.5— 2.8wt. %的缩水甘 油醚类交联剂，优选 l-2.5wt.%， 更优选 l-2wt.%,反应温度 40-60°C， 优选 45-55 °C , 反应时间 1-10小时， 放置冷却；  The sodium hyaluronate is dissolved in a sodium hydroxide solution having a concentration of 0.3% to 0.9%, the concentration of hyaluronic acid is 6%-10%, preferably 6%-8%, and the weight of sodium hyaluronate is 0.5-2.8 wt. % glycidyl ether type crosslinking agent, preferably 1-2.5 wt.%, more preferably 1-2 wt.%, reaction temperature 40-60 ° C, preferably 45-55 ° C, reaction time 1-10 hours, placed cool down;
(2)连接反应  (2) Connection reaction
将步骤 （1 ) 的反应物， 加入一次交联反应中相同重量、 相同浓度的氢 氧化纳溶液搅匀；  The reactant of the step (1) is added to the same weight and the same concentration of the sodium hydroxide solution in the primary crosslinking reaction;
(3 ) 二次交联  (3) secondary cross-linking
将步骤 （2) 的反应物加入与 （1 ) 相同或不同的缩水甘油醚类交联剂， 其加入量为透明质酸钠重量的 3-7wt.%, 优选 3.75-6.25 t%，更优选 3-5wt.% b 充分搅拌， 使溶液均匀。开始加热， 反应时间 1-5小时， 反应温度为 40-60°C， 优选 50-60°C， 更优选 50-53 °C， 凝胶形成；  The reactant of the step (2) is added to the same or different glycidyl ether type crosslinking agent as (1), and the amount thereof is 3-7 wt.%, preferably 3.75-6.25 t%, more preferably the weight of the sodium hyaluronate. 3-5 wt.% b Stir well to make the solution uniform. Heating is started, the reaction time is 1-5 hours, the reaction temperature is 40-60 ° C, preferably 50-60 ° C, more preferably 50-53 ° C, gel formation;
(4)将步骤（3 )的凝胶放入浓度为 0.8-1%的 NaCl溶液中浸泡， 并将其 用 HC1溶液中和至 PH值为 6.5-7.5。  (4) The gel of the step (3) is immersed in a NaCl solution having a concentration of 0.8-1%, and neutralized with a HC1 solution to a pH of 6.5-7.5.
该制备方法中， 所述步骤 （4) 中的浸泡是将凝胶浸泡至透明质酸的含量 为凝胶总重量的 1一 3wt%，优选 l-2%w 。 并且该制备方法还包括当凝胶浸泡 至透明质酸的含量为凝胶总重量的 1一 3wt. %时，进行粉碎的步骤,更优选当凝 胶浸泡至透明质酸的含量为凝胶总重量的 i一 3wt. %时， 进行粉碎的步骤， 该制备方法中， 所述的缩水甘油醚类交联剂为乙二醇二缩水甘油醚和 /或 1， 4一丁二醇缩水甘油醚。  In the preparation method, the soaking in the step (4) is to soak the gel to a content of hyaluronic acid of 1 to 3 wt%, preferably 1 to 2% w, based on the total weight of the gel. And the preparation method further comprises the step of pulverizing when the gel is soaked to a content of hyaluronic acid of 1 to 3 wt.% of the total weight of the gel, more preferably when the gel is soaked to a content of hyaluronic acid as a total gel. When the weight is i3 wt.%, a pulverization step is carried out, wherein the glycidyl ether type crosslinking agent is ethylene glycol diglycidyl ether and/or 1,4-butanediol glycidyl ether. .
该制备方法中， 步骤 （4) 中所述的 NaCl溶液的质量浓度为 0.8-1%; 所 述 HC1的质量浓度为 0.3-0.9%。  In the preparation method, the mass concentration of the NaCl solution described in the step (4) is 0.8-1%; and the mass concentration of the HC1 is 0.3-0.9%.
一种透明质酸凝胶， 该透明质酸凝胶是由上述制备方法制备的。  A hyaluronic acid gel prepared by the above preparation method.
一种医用材料， 包括除皱衬垫原料， 该除皱衬垫原料是上述透明质酸凝 胶浸泡、 中和后， 经粉碎后形成的。  A medical material, comprising a wrinkle pad material, which is formed by immersing and immersing the hyaluronic acid gel after the above-mentioned hyaluronic acid gel.
该医用材料中，所述的浸泡是在 NaCi溶液中浸泡，所述的中和是用 HCL 溶液中和至 PH为 6.5-7.5。 In the medical material, the soaking is immersed in a NaCi solution, and the neutralization is performed by HCL. The solution was neutralized to a pH of 6.5-7.5.
其中，所述的 NaCl溶液的质量浓度为 0.8-1%%; 所述 HC1的质量浓度为 0.3-0.9%。  Wherein, the mass concentration of the NaCl solution is 0.8-1%%; and the mass concentration of the HC1 is 0.3-0.9%.
该医用材料中， 所述的透明质酸凝胶浸泡后透明质酸的含量为凝胶总重 量的 1一 3wt。/0。 本发明的透明质酸凝胶， 是通过二次交联完成的， 可以使交联剂的用量显 著的降低， 并且减少了交联剂的残留， 最大程度地减少交联剂的残留对注射 部位及人体的影响， 有效防止的交联剂对人体的损害。 同时， 有效地提高了 透明质酸凝胶的保存时间， 使其在可注射部位提高了保留性， 增长保留时间， 提高凝胶品质。 因此， 本发明的透明质酸凝胶可以作为除皱的衬垫材料， 用 于医疗或美容行业。 具体实施方式 以下结合实施例对本发明作详细的说明： 实施例 1 取透明质酸钠 8g, 溶于浓度为 0.8% 的氢氧化钠溶液 100g中， 透明质酸 的浓度约 8%， 加入 1.4-丁二醇缩水甘油醚 0.08g， 于 45°C下加热 3小时。 放 置， 再加入上述同重量、 同浓度的氢氧化纳溶液。然后再加入 1.4-丁二醇缩水 甘油醚 0.5g， 充分搅拌， 在温度 50°C下加热， 加热时间 2.5小时， 制得透明 质酸凝胶。 In the medical material, the content of hyaluronic acid after soaking the hyaluronic acid gel is 1-3 wt% of the total weight of the gel. / 0 . The hyaluronic acid gel of the present invention is completed by secondary crosslinking, and the amount of the crosslinking agent can be remarkably reduced, and the residue of the crosslinking agent can be reduced, and the residue of the crosslinking agent can be minimized to the injection site. And the influence of the human body, effectively preventing the damage of the cross-linking agent to the human body. At the same time, the preservation time of the hyaluronic acid gel is effectively improved, which improves the retention at the injectable site, increases the retention time, and improves the gel quality. Therefore, the hyaluronic acid gel of the present invention can be used as a cushioning material for wrinkles for use in the medical or cosmetic industry. BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below with reference to examples: Example 1 8 g of sodium hyaluronate was dissolved in 100 g of a sodium hydroxide solution having a concentration of 0.8%, and the concentration of hyaluronic acid was about 8%, and 1.4- 0.08 g of butanediol glycidyl ether was heated at 45 ° C for 3 hours. Place and add the above-mentioned sodium hydroxide solution of the same weight and the same concentration. Then, 0.5 g of 1.4-butylene glycol glycidyl ether was further added, stirred well, and heated at a temperature of 50 ° C for 2.5 hours to prepare a hyaluronic acid gel.
将上述制备的透明质酸凝胶在浓度为 0.9%的 NaCl溶液中浸泡，并用浓度 为 0.3% 的 HC1溶液中和至 PH值为 6.5，凝胶浸泡至透明质酸含量为 1%。粉 碎为相应颗粒， 即为理想的除皱衬垫原料。 将该透明质酸凝胶进行透明质酸 酶的降解实验， 并与使用相同透明质酸及交联剂的一次交联的凝胶颗粒进行 比较， 结果， 二次交联的凝胶颗粒降解时间为一次交联的 20.92倍。 实施例 2 取透明质酸钠 8g， 溶于浓度为 0.6% 的氢氧化钠溶液 120g中， 透明质酸 的浓度约 6.7%, 加入 0.2g乙二醇縮水甘油醚， 于 50°C下加热 2小时。 放置， 再加入上述同重量、 同浓度的氢氧化钠溶液。 然后再加入 0.3g乙二醇縮水甘 油醚交联剂， 充分搅拌， 在 53°C下加热， 加热时间为 1.5小时， 制得透明质 酸凝胶。 The hyaluronic acid gel prepared above was immersed in a 0.9% NaCl solution and neutralized to a pH of 6.5 with a 0.3% HCl solution, and the gel was soaked to a hyaluronic acid content of 1%. The pulverization into the corresponding granules is an ideal crease liner material. The hyaluronic acid gel was subjected to hyaluronidase degradation experiments and compared with primary crosslinked gel particles using the same hyaluronic acid and a crosslinking agent, and as a result, the secondary crosslinked gel particles were degraded. 20.92 times for one cross-linking. Example 2 8 g of sodium hyaluronate was dissolved in 120 g of a sodium hydroxide solution having a concentration of 0.6%, and the concentration of hyaluronic acid was about 6.7%. 0.2 g of ethylene glycol glycidyl ether was added thereto, and the mixture was heated at 50 ° C for 2 hours. Place and add the above-mentioned sodium hydroxide solution of the same weight and the same concentration. Then, 0.3 g of an ethylene glycol glycidyl ether crosslinking agent was further added, and the mixture was thoroughly stirred, and heated at 53 ° C for 1.5 hours to prepare a hyaluronic acid gel.
将上述制备的透明质酸凝胶在浓度为 0.8%的 NaCl溶液中浸泡， 并用浓 度为 0.6% 的 HC1溶液中和至 PH值为 7， 凝胶浸泡至透明质酸含量为 2%。 粉碎为相应颗粒， 即为理想的除皱衬垫原料。 将该透明质酸凝胶进行透明质 酸酶的降解实验， 并与使用相同透明质酸及交联剂的一次交联的凝胶颗粒进 行比较， 结果， 二次交联的凝胶颗粒降解时间为一次交联的 46.65倍。 实施例 3 取透明质酸钠 8g， 溶于浓度为 0.3% 的氢氧化钠溶液 90g中， 透明质酸 的浓度约 8.9%，加入 0.15g的 1.4-丁二醇縮水甘油醚。于 55°C下加热 2小时， 放置。 然后再加入同重量、 同浓度的氢氧化钠溶液， 充分搅拌， 然后再加入 0.43g乙二醇縮水甘油醚交联剂， 充分搅拌， 在温度 55°C下反应 1.5小时， 制 得透明质酸凝胶。  The hyaluronic acid gel prepared above was immersed in a 0.8% NaCl solution, and neutralized to a pH of 7, with a concentration of 0.6% of the HC1 solution, soaked to a hyaluronic acid content of 2%. The pulverization into the corresponding granules is an ideal crease liner material. The hyaluronic acid gel was subjected to hyaluronidase degradation experiments and compared with primary crosslinked gel particles using the same hyaluronic acid and a crosslinking agent, and as a result, the secondary crosslinked gel particles were degraded. 46.65 times as much as one cross-linking. Example 3 8 g of sodium hyaluronate was dissolved in 90 g of a sodium hydroxide solution having a concentration of 0.3%, and the concentration of hyaluronic acid was about 8.9%, and 0.15 g of 1.4-butylene glycol glycidyl ether was added. Heat at 55 ° C for 2 hours and place. Then add the same weight, the same concentration of sodium hydroxide solution, stir well, then add 0.43g ethylene glycol glycidyl ether crosslinker, stir well, react at temperature 55 ° C for 1.5 hours, to produce hyaluronic acid gel.
将上述制备的透明质酸凝胶在浓度为 1% 的 NaCl溶液中浸泡，并用浓度 为 0.9% 的 HC1溶液中和至 PH值为 7.5，凝胶浸泡至透明质酸含量为 3%。粉 碎为相应颗粒， 即为理想的除皱衬垫原料。 将该透明质酸凝胶进行透明质酸 酶的降解实验， 并与使用相同透明质酸及交联剂的一次交联的凝胶颗粒进行 比较， 结果， 二次交联的凝胶颗粒降解时间为一次交联的 10.88倍。  The hyaluronic acid gel prepared above was immersed in a NaCl solution having a concentration of 1%, and neutralized to a pH of 7.5 with a 0.9% HC1 solution, and the gel was soaked to a hyaluronic acid content of 3%. The powder is broken into the corresponding particles, which is the ideal wrinkle pad material. The hyaluronic acid gel was subjected to hyaluronidase degradation experiments and compared with primary crosslinked gel particles using the same hyaluronic acid and a crosslinking agent, and as a result, the secondary crosslinked gel particles were degraded. 10.88 times for one cross-linking.
实验中采用不同的一次， 二次交联剂的配比量， 得到不同性能的凝胶， 浸 泡， 中和后， 使透明质酸含量大约为 1-3%下进行透明质酸酶的降解实验。 实 验结果，可以得到新的透明质酸凝胶，其降解时间为一次胶联凝胶的 2-50倍。 并且降解时间相同时， 二次交联的交联剂加入量竟为一次交联的交联剂加入 量的 60%。  In the experiment, different ratios of secondary cross-linking agents were used to obtain gels with different properties. After soaking and neutralizing, hyaluronic acid degradation experiments were carried out with hyaluronic acid content of about 1-3%. . As a result of the experiment, a new hyaluronic acid gel having a degradation time of 2 to 50 times that of the primary gelled gel can be obtained. When the degradation time is the same, the cross-linking agent is added in an amount of 60% of the cross-linking cross-linking agent.
值得注意的是，本实施例仅以乙二醇縮水甘油醚交联剂和 1.4-丁二醇缩水 甘油醚为例对本发明技术方案的具体工艺进行了说明， 但是， 根据本领域的 技术可知， 本领域的技术人员显然易见地会认识到任何缩水甘油醚类交联剂 均能实现该技术方案并能达到本发明的技术目的， 该透明质酸的制备方法对 任何缩水甘油醚类交联剂来说是普适的， 采用任何缩水甘油醚作交联剂的方 案显然能得到本发明公幵内容的有力支持。 It is worth noting that this example only uses ethylene glycol glycidyl ether crosslinker and 1.4-butanediol shrinkage. The specific process of the technical solution of the present invention is described by taking glyceryl ether as an example. However, it will be apparent to those skilled in the art that any glycidyl ether type crosslinking agent can realize the technical solution according to the art. To achieve the technical object of the present invention, the preparation method of the hyaluronic acid is universal for any glycidyl ether type crosslinking agent, and the use of any glycidyl ether as a crosslinking agent can obviously obtain the contents of the present invention. Strong support.
尽管上文对本发明的实施例进行了详细的描述和说明， 但应该指明的是， 我们可以对上述实施例进行各种改变和修改， 但这些都不脱离本发明的精神 和所附的权利要求所记载的范围。  While the embodiments of the present invention have been described and illustrated in the foregoing embodiments of the present invention The range described.

Claims

权 利 要 求 书 Claim
1、 一种透明质酸凝胶的制备方法， 包括以下步骤： A method for preparing a hyaluronic acid gel, comprising the steps of:
( 1 ) 一次交联  (1) One cross-linking
将透明质酸钠溶于浓度 0.3 %— 0.9%的氢氧化钠溶液中， 透明质酸的浓 度为 6%— 10%， 加入透明质酸钠重量的 0.5— 3wt.%的縮水甘油醚类交联剂， 反应温度 40-60°C, 反应时间 1-10小时,放置冷却；  The sodium hyaluronate is dissolved in a sodium hydroxide solution having a concentration of 0.3% to 0.9%, the concentration of hyaluronic acid is 6%-10%, and the weight of the sodium hyaluronate is 0.5-3 wt.% of the glycidyl ether. The crosslinking agent, the reaction temperature is 40-60 ° C, the reaction time is 1-10 hours, and it is left to cool;
(2) 连接反应  (2) Connection reaction
将步骤 （1 ) 的反应物， 加入一次交联反应中相同重量、 相同浓度的氢 氧化纳溶液搅匀；  The reactant of the step (1) is added to the same weight and the same concentration of the sodium hydroxide solution in the primary crosslinking reaction;
(3 ) 二次交联  (3) secondary cross-linking
将步骤 （2) 的反应物加入与 （1 ) 相同或不同的缩水甘油醚类交联剂， 其加入量为透明质酸钠重量的 3-7wt.%。 充分搅拌， 使溶液均匀。 开始加热， 反应时间 1-5小时， 温度 40-60°C， 凝胶形成；  The reactant of the step (2) is added to the same or different glycidyl ether type crosslinking agent as (1) in an amount of from 3 to 7 wt.% based on the weight of the sodium hyaluronate. Stir well to make the solution even. Start heating, reaction time 1-5 hours, temperature 40-60 ° C, gel formation;
(4)将步骤（3 ) 的凝胶放入浓度为 0.8-1%的 NaCl溶液中浸泡， 并将其 用 HC1溶液中和至 PH值为 6.5-7.5。  (4) The gel of the step (3) is immersed in a NaCl solution having a concentration of 0.8-1%, and neutralized with a HC1 solution to a pH of 6.5-7.5.
2、 如权利要求 1所述的透明质酸凝胶的制备方法， 其特征在于， 所述步 骤（4) 中的浸泡是将凝胶浸泡至透明质酸的含量为凝胶总重量的 1一 3wt.%。  The method for preparing a hyaluronic acid gel according to claim 1, wherein the immersing in the step (4) is: soaking the gel to a content of hyaluronic acid of 1% of the total weight of the gel. 3wt.%.
3、如权利要求 2所述的透明质酸凝胶的制备方法，其特征在于，步骤（4) 的浸泡后还包括粉碎的步骤。  The method for preparing a hyaluronic acid gel according to claim 2, characterized in that the step of pulverizing after the immersing in the step (4).
4、 如权利要求 1-3任一项所述的透明质酸凝胶的制备方法， 其特征在于， 所述的缩水甘油醚类交联剂为乙二醇二缩水甘油醚和 /或 1， 4一丁二醇缩水甘 油醚。  The method for producing a hyaluronic acid gel according to any one of claims 1 to 3, wherein the glycidyl ether type crosslinking agent is ethylene glycol diglycidyl ether and/or 1, 4-butylene glycol glycidyl ether.
5、如权利要求 4所述的透明质酸凝胶的制备方法，其特征在于，步骤（4) 中所述的 NaCl溶液的浓度为 0.8%-1%; 所述 HC1的浓度为 0.3%-0.9%。 The method for preparing a hyaluronic acid gel according to claim 4, wherein the concentration of the NaCl solution in the step (4) is 0.8% to 1% ; and the concentration of the HC1 is 0.3%. 0.9%.
6、如权利要求 1-3任一项所述的透明质酸凝胶的制备方法，其特征在于, 步骤 （4) 中所述 NaCl 溶液的浓度为 0.8 %-1%; 所述 HC1 的质量浓度为 0.3%-0.9%0 The method for preparing a hyaluronic acid gel according to any one of claims 1 to 3, wherein the concentration of the NaCl solution in the step (4) is from 0.8% to 1% ; the quality of the HC1 Concentration is 0.3%-0.9% 0
7、 一种透明质酸凝胶， 该透明质酸凝胶是由上述权利要求 1-6所述的任 一项制备方法制备的。 A hyaluronic acid gel prepared by the production method according to any one of claims 1 to 6 above.
8、 一种医用材料， 包括除皱衬垫原料， 该除皱衬垫原料是上述权利要求 1-6任一项所述的透明质酸凝胶制备方法所制备的透明质酸凝胶，浸泡、 中和、 并经粉碎后形成的。 A medical material, comprising a wrinkle pad material, the wrinkle pad material is a hyaluronic acid gel prepared by the method for preparing a hyaluronic acid gel according to any one of claims 1 to 6, soaking , neutralized, and formed after crushing.
9、 如权利要求 8所述的医用材料， 其特征在于， 所述的浸泡是在 NaCl 溶液中浸泡，其浓度为 0.8%-1%; ; 所述的中和是用 HC1 溶液中和至 PH为 6.5-7.5, 其浓度为 0.3%-0.9%。 9. The medical material according to claim 8, wherein the immersion is immersed in a NaCl solution at a concentration of 0.8% to 1% ; and the neutralization is neutralized to a pH with an HC1 solution. It is 6.5-7.5 and its concentration is 0.3%-0.9%.
10、如权利要求 8-9所述的医用材料， 其特征在于， 所述的透明质酸凝胶 浸泡后， 透明质酸的含量为凝胶总重量的 1一 3wt.%。  The medical material according to any one of claims 8-9, wherein the hyaluronic acid gel has a hyaluronic acid content of 1-3 wt.% of the total weight of the gel.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110964215A (en) * 2019-12-26 2020-04-07 华熙生物科技股份有限公司 Preparation method of L-polylactic acid and cross-linked hyaluronic acid composite gel for injection and obtained product
EP2649101B1 (en) 2010-12-06 2021-06-02 Teoxane Process of preparing a cross linked gel

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101821294B (en) * 2007-09-28 2012-10-31 株式会社资生堂 Swellable crosslinked hyaluronic acid powder and method for producing the same
CN101724164B (en) * 2008-10-31 2011-12-14 科妍生物科技股份有限公司 Method for preparing cross-linked hyaluronic acid
CN101829365B (en) * 2010-04-13 2013-04-17 杭州市第一人民医院 Sclera external pressurized biologic composite materials and preparation method
CN102757570A (en) * 2011-04-25 2012-10-31 颜先琴 Preparation method of sodium hyaluronate gel
CN103146003A (en) * 2013-03-06 2013-06-12 上海其胜生物制剂有限公司 Preparation method of low-temperature secondary cross-linked sodium hyaluronate gel
CA2916330A1 (en) * 2013-06-28 2014-12-31 Galderma S.A. A process for preparing a cross-linked hyaluronic acid product
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CN113087935B (en) * 2021-05-19 2022-05-27 青岛琛蓝海洋生物工程有限公司 Composite sodium hyaluronate gel for resisting hyaluronidase hydrolysis and preparation method thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4716224A (en) * 1984-05-04 1987-12-29 Seikagaku Kogyo Co. Ltd. Crosslinked hyaluronic acid and its use
CN1342171A (en) * 1999-02-03 2002-03-27 维特罗莱夫英国有限公司 Process for production of multiple cross-linked hyaluronic acid derivatives
US20040127698A1 (en) * 2002-12-31 2004-07-01 Industrial Technology Research Institute Method for producing double-crosslinked hyaluronate material

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4716224A (en) * 1984-05-04 1987-12-29 Seikagaku Kogyo Co. Ltd. Crosslinked hyaluronic acid and its use
CN1342171A (en) * 1999-02-03 2002-03-27 维特罗莱夫英国有限公司 Process for production of multiple cross-linked hyaluronic acid derivatives
US20040127698A1 (en) * 2002-12-31 2004-07-01 Industrial Technology Research Institute Method for producing double-crosslinked hyaluronate material

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2649101B1 (en) 2010-12-06 2021-06-02 Teoxane Process of preparing a cross linked gel
CN110964215A (en) * 2019-12-26 2020-04-07 华熙生物科技股份有限公司 Preparation method of L-polylactic acid and cross-linked hyaluronic acid composite gel for injection and obtained product
CN110964215B (en) * 2019-12-26 2022-03-29 华熙生物科技股份有限公司 Preparation method of L-polylactic acid and cross-linked hyaluronic acid composite gel for injection and obtained product

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