WO2017117996A1 - 一种外科止血生物制品及其使用方法 - Google Patents

一种外科止血生物制品及其使用方法 Download PDF

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WO2017117996A1
WO2017117996A1 PCT/CN2016/092996 CN2016092996W WO2017117996A1 WO 2017117996 A1 WO2017117996 A1 WO 2017117996A1 CN 2016092996 W CN2016092996 W CN 2016092996W WO 2017117996 A1 WO2017117996 A1 WO 2017117996A1
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collagen
lyophilized powder
biological product
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张俊辉
朱晋辉
万华印
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广州市众为生物技术有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0028—Polypeptides; Proteins; Degradation products thereof
    • A61L26/0042—Fibrin; Fibrinogen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0028—Polypeptides; Proteins; Degradation products thereof
    • A61L26/0033—Collagen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0028—Polypeptides; Proteins; Degradation products thereof
    • A61L26/0047—Specific proteins or polypeptides not covered by groups A61L26/0033 - A61L26/0042
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0061—Use of materials characterised by their function or physical properties
    • A61L26/0066—Medicaments; Biocides
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L89/00—Compositions of proteins; Compositions of derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412—Tissue-regenerating or healing or proliferative agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising agents

Definitions

  • the invention relates to the technical fields of biopharmaceuticals, biological products, biological materials and medical instruments, in particular to an efficient surgical hemostatic biological product, which can be used for hemostasis, adhesion and sealing in surgery.
  • Rapid and complete hemostasis during surgery is one of the core operations of the basic operation of surgery.
  • various kinds of first-aid such as frequent traffic accidents and accidents, it is also generally faced with problems such as rapid and effective hemostasis, alleviating patient suffering and saving people's lives.
  • the technical problem to be solved by the present invention is to provide a surgical hemostatic biological product and a method for using the same, which accelerates wound healing.
  • a surgical hemostatic biological product comprising a first lyophilized powder and a second lyophilized powder, the components of the first lyophilized powder comprising: fibrinogen, XIII Coagulation factor and microfibrous collagen; components of the second lyophilized powder include: thrombin and microfibrous collagen.
  • Fibrinogen is hydrolyzed to fibrin monomer under the action of thrombin, and then cross-linked to form a solidizable fibrin solid, thereby adhering to the surface of the bleeding wound and functioning to stop bleeding.
  • Microfibrous collagen is an absorbable local hemostatic material made of bovine dermis. It adheres to the bleeding wound and accumulates platelets to form clots to quickly stop bleeding. Microfibrous collagen is also an animal-derived protein that can be completely absorbed by the body.
  • the fibrinogen, XIII coagulation factor and thrombin are derived from a mammal.
  • a method for using a surgical hemostatic biological product comprising the following steps:
  • the content of the microfiber collagen in the first solution is 0.1 to 20 mg/ml
  • the content of the microfiber collagen in the second solution is 0.1 to 20 mg/ml
  • the content of the microfiber collagen in the first solution is 0.2 to 10 mg/ml
  • the content of the microfiber collagen in the second solution is 0.2 to 10 mg/ml
  • the content of the microfiber collagen in the first solution is 0.5 to 5 mg/ml
  • the content of the microfiber collagen in the second solution is 0.5 to 5 mg/ml
  • fibrin and microfibrillar collagen II can be exerted.
  • the coagulation effect makes the hemostasis faster and the wound healing effect is better after use.
  • the human fibrinogen intermediate was dissolved in an appropriate amount of sodium chloride (3 g/L, sodium citrate 1.5 g/L, sodium hydrogencarbonate 0.8 g/L, L-arginine hydrochloride 3.3 g/L).
  • the protein concentration was 4%, and then the microfibrous collagen was added to 3 g/L, the pH was adjusted to 7.2, and the mixture was centrifuged and lyophilized to obtain the first lyophilized powder.
  • the human thrombin intermediate was formulated into 550 IU/ml, glycine 5 g/L, human albumin 2 g/L, microfiber collagen 12 g/L, centrifuged, and lyophilized to obtain a second lyophilized powder.
  • the adhesion strength was measured to be greater than 460 gf/cm 2 according to the spray area of 10 cm 2 /ml.
  • the wounds were 0.2cm deep, 1.0cm long and 0.5cm wide. Ten of them were treated with fibrin glue without microfibrous collagen. Ten of them were hemostasis with microfiber collagen. . The proportion of rats that stopped bleeding in 1 minute was 45% for the former and 90% for the latter. This product containing microfibrous collagen has a better hemostatic effect.
  • the porcine fibrinogen intermediate was dissolved in the protein with an appropriate amount of sodium chloride (3 g/L, sodium citrate 2 g/L, polysorbate 800.2 g/L, L-arginine hydrochloride 3.3 g/L). The concentration was 3%, and then the microfibrous collagen was added to 4.5 g/L, the pH was adjusted to 7.5, and the mixture was centrifuged, and then lyophilized, which was the first lyophilized powder.
  • the porcine thrombin intermediate was formulated into 550 IU/ml, glycine 5g/L, human albumin 2g/L, microfibrous collagen 12.5g/L, centrifuged and lyophilized, which was the second lyophilized powder. .
  • the porcine fibrinogen intermediate was dissolved in the protein with an appropriate amount of sodium chloride (2 g/L, sodium citrate 2 g/L, polysorbate 800.2 g/L, L-arginine hydrochloride 3.3 g/L). The concentration was 3.5%, and then the thiol-microfiber collagen was added to 2.5 g/L, the pH was adjusted to 7.5, and the mixture was centrifuged and lyophilized to obtain the first lyophilized powder.
  • the pig-derived thrombin intermediate was formulated into 550 IU/ml, glycine 15 g/L, human albumin 2 g/L, and microfiber collagen 1 g/L. After centrifugation and filtration, the lyophilized product is divided into two lyophilized powders.

Abstract

一种外科止血生物制品及其使用方法，外科止血生物制品包括第一冻干粉和第二冻干粉，第一冻干粉的组分包括纤维蛋白原、XIII凝血因子和微纤维胶原；第二冻干粉的组分包括凝血酶和微纤维胶原。使用时，将两种冻干粉溶解在溶解液中，然后喷洒在创面，加速伤口的愈合。

Description

一种外科止血生物制品及其使用方法 技术领域
本发明是涉及生物制药、生物制品、生物材料及医疗器械等技术领域，具体是一种高效外科止血生物制品，该产品可用于外科手术中的止血、粘合和封闭。
背景技术
手术中迅速彻底的止血，减少出血量，保持术野清晰，是外科手术基本操作的核心之一。在各种急救如频繁发生的交通及意外事故中，也普遍面临快速而有效止血，减轻病人痛苦，挽救人民生命等问题。
局部止血材料广泛用于外科止血，其止血效果在动物实验和临床中都得到了充分肯定。使用局部止血药不仅可以减少出血量,简化手术操作，缩短手术时间，有时还可促进伤口的愈合。
目前,常用的局部止血药有纤维蛋白胶、明胶海绵、壳聚糖、氧化纤维素、微纤维胶原、沸石粉、凝血酶及海藻酸纤维等。各种不同类型的局部止血药，由于它们的结构和组成成分不同，其理化性质也不一样,所以,它们的作用机理和使用方法不尽相同，止血作用效果也有很大差异。
纤维蛋白(原)作为医用粘合剂的历史可以追溯至上世纪初，1909年Bergel报道纤维蛋白粉末具有止血功能，Grey随即用它来进行肝脏和大脑的止血。1940年，Young尝试利用凝血反应来粘合切断的周围神经，Tidrick和Cronkile等则用它来固定移植的皮肤。但纤维蛋白作为粘合剂是直到1972年才由Matras确立的。Matras首次用冷沉淀技术提取了高浓度的人纤维蛋白原，加上高浓度牛凝血酶和第XIII凝血因子制成粘合剂，以之进行周围神经吻合获得成功。纤维蛋白原在凝血酶作用下水解成纤维蛋白单体，再交联形成可凝固的纤维蛋白固体，从而粘附在出血创伤表面，起到止血的功效。
发明内容
本发明所要解决的技术问题是提供一种外科止血生物制品及其使用方法，加速 伤口愈合。
为解决上述技术问题，本发明的技术方案是：一种外科止血生物制品，包括第一冻干粉和第二冻干粉，所述第一冻干粉的组分包括：纤维蛋白原、XIII凝血因子和微纤维胶原；所述第二冻干粉的组分包括：凝血酶和微纤维胶原。
纤维蛋白(原)作为医用粘合剂的历史可以追溯至上世纪初，1909年Bergel报道纤维蛋白粉末具有止血功能，Grey随即用它来进行肝脏和大脑的止血。1940年，Young尝试利用凝血反应来粘合切断的周围神经，Tidrick和Cronkile等则用它来固定移植的皮肤。但纤维蛋白作为粘合剂是直到1972年才由Matras确立的。Matras首次用冷沉淀技术提取了高浓度的人纤维蛋白原，加上高浓度牛凝血酶和第XIII凝血因子制成粘合剂，以之进行周围神经吻合获得成功。纤维蛋白原在凝血酶作用下水解成纤维蛋白单体，再交联形成可凝固的纤维蛋白固体，从而粘附在出血创伤表面，起到止血的功效。微纤维胶原是由牛真皮制成的可吸收性局部止血材料，在出血创面黏附、聚集血小板形成凝血块迅速止血。微纤维胶原也是来源于动物的蛋白质，其可以被机体完全吸收。
作为改进，所述纤维蛋白原、XIII凝血因子和凝血酶来源于哺乳动物。
为解决上述技术问题，本发明的技术方案的使用方法：一种外科止血生物制品的使用方法，包括以下步骤：
(1)将第一冻干粉溶解在第一溶解液中，将第二冻干粉溶解在第二溶解液中；
(2)将第一溶解液和第二溶解液混合并喷洒到创面；
(3)混合液用于止血、粘合和封闭。
作为改进，所述第一溶解液中，微纤维胶原的含量为0.1～20mg/ml，所述第二溶解液中，微纤维胶原的含量为0.1～20mg/ml。
作为改进，所述第一溶解液中，微纤维胶原的含量为0.2～10mg/ml，所述第二溶解液中，微纤维胶原的含量为0.2～10mg/ml。
作为改进，所述第一溶解液中，微纤维胶原的含量为0.5～5mg/ml，所述第二溶解液中，微纤维胶原的含量为0.5～5mg/ml。
本发明与现有技术相比所带来的有益效果是：
在蛋白胶类止血生物制品中添加微纤维胶原后，可发挥纤维蛋白和微纤维胶原二 者的凝血作用，使止血速度更快，使用后的伤口愈合效果更好。
具体实施方式
实施例1
将人源纤维蛋白原中间体用适量溶解液(氯化钠3g/L，枸橼酸钠1.5g/L，碳酸氢钠0.8g/L，L-盐酸精氨酸3.3g/L)溶解至蛋白浓度为4％，再加入微纤维胶原至3g/L，调pH为7.2，离心过滤后分装冻干，即为第一冻干粉。将人源凝血酶中间体配制成550IU/ml，加入甘氨酸5g/L，人血白蛋白2g/L，微纤维胶原12g/L，离心过滤后分装冻干，即为第二冻干粉。
用溶解液把两种冻干粉溶解后，使用配药器喷洒，按喷洒面积10cm2/ml规格，检测其粘合强度大于460gf/cm2。对20只大鼠肝表面造深0.2cm、长1.0cm、宽0.5cm的创面，其中10只采用普通未添加微纤维胶原的纤维蛋白胶止血，10只采用添加了微纤维胶原的本产品止血。在1分钟内止血的大鼠比例，前者为45％，后者为90％。含有微纤维胶原的本产品具有更好的止血效果。
实施例2
将猪源纤维蛋白原中间体用适量溶解液(氯化钠3g/L，枸橼酸钠2g/L，聚山梨酯800.2g/L，L-盐酸精氨酸3.3g/L)溶解至蛋白浓度为3％，再加入微纤维胶原至4.5g/L，调pH为7.5，离心过滤后分装冻干，即为第一冻干粉。将猪源凝血酶中间体配制成550IU/ml，加入甘氨酸5g/L，人血白蛋白2g/L，微纤维胶原12.5g/L，离心过滤后分装冻干，即为第二冻干粉。
用溶解液把两种冻干粉溶解后，使用配药器喷洒，按喷洒面积10cm2/ml规格，检测其粘合强度大于250gf/cm2。
实施例3
将猪源纤维蛋白原中间体用适量溶解液(氯化钠2g/L，枸橼酸钠2g/L，聚山梨酯800.2g/L，L-盐酸精氨酸3.3g/L)溶解至蛋白浓度为3.5％，再加入巯基-微纤维胶原至2.5g/L，调pH为7.5，离心过滤后分装冻干，即为第一冻干粉。将猪源凝血酶中间体配制成550IU/ml，加入甘氨酸15g/L，人血白蛋白2g/L，微纤维胶原1g/L， 离心过滤后分装冻干，即为第二冻干粉。
用溶解液把两种冻干粉溶解后，使用配药器喷洒，按喷洒面积10cm2/ml规格，检测其粘合强度大于250gf/cm2，弹性高度7.1cm。

Claims (6)

  1. 一种外科止血生物制品，其特征在于：包括第一冻干粉和第二冻干粉，所述第一冻干粉的组分包括：纤维蛋白原、XIII凝血因子和微纤维胶原；
    所述第二冻干粉的组分包括：凝血酶和微纤维胶原。
  2. 根据权利要求1所述的一种外科止血生物制品，其特征在于：所述纤维蛋白原、XIII凝血因子和凝血酶来源于哺乳动物。
  3. 一种如权利要求1所述的一种外科止血生物制品的使用方法，其特征在于，包括以下步骤：
    (1)将第一冻干粉溶解在第一溶解液中，将第二冻干粉溶解在第二溶解液中；
    (2)将第一溶解液和第二溶解液混合并喷洒到创面；
    (3)混合液用于止血、粘合和封闭。
  4. 根据权利要求3所述的一种外科止血生物制品的使用方法，其特征在于：所述第一溶解液中，微纤维胶原的含量为0.1～20mg/ml，所述第二溶解液中，微纤维胶原的含量为0.1～20mg/ml。
  5. 根据权利要求3所述的一种外科止血生物制品的使用方法，其特征在于：所述第一溶解液中，微纤维胶原的含量为0.2～10mg/ml，所述第二溶解液中，微纤维胶原的含量为0.2～10mg/ml。
  6. 根据权利要求3所述的一种外科止血生物制品的使用方法，其特征在于：所述第一溶解液中，微纤维胶原的含量为0.5～5mg/ml，所述第二溶解液中，微纤维胶原的含量为0.5～5mg/ml。
PCT/CN2016/092996 2016-01-08 2016-08-03 一种外科止血生物制品及其使用方法 WO2017117996A1 (zh)

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