WO2017045377A1 - Dll4细胞因子在制备治疗暴发性肝功能衰竭药剂中的应用 - Google Patents

Dll4细胞因子在制备治疗暴发性肝功能衰竭药剂中的应用 Download PDF

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WO2017045377A1
WO2017045377A1 PCT/CN2016/075574 CN2016075574W WO2017045377A1 WO 2017045377 A1 WO2017045377 A1 WO 2017045377A1 CN 2016075574 W CN2016075574 W CN 2016075574W WO 2017045377 A1 WO2017045377 A1 WO 2017045377A1
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dll4
treatment
cytokines
control group
fulminant hepatic
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PCT/CN2016/075574
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English (en)
French (fr)
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李君�
李兰娟
陈新
李江
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浙江大学
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Priority to US15/367,239 priority Critical patent/US10328122B2/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons

Definitions

  • the invention belongs to the fields of clinical medicine, molecular medicine and regenerative medicine, in particular, is a new technology of DLL4 cytokine series drugs for treating liver failure.
  • Fulminant liver failure is a kind of disease caused by a large area of liver necrosis caused by various reasons.
  • the condition is dangerous, the patient is more than the short-term death, and the conventional medical comprehensive treatment has a mortality rate of more than 80%.
  • the conventional medical comprehensive treatment has a mortality rate of more than 80%.
  • there is currently no specific treatment In addition to orthotopic liver transplantation, there is currently no specific treatment. However, due to a serious shortage of donor liver, a large number of patients died while waiting for liver transplantation. If one or more cytokines that inhibit liver necrosis, rapidly induce host liver tissue regeneration, and long-term function, and avoid potential side effects, can be found for the treatment of fulminant hepatic failure caused by various causes. Extremely important.
  • the DLL4 cytokine is a type I transmembrane protein located on the cell membrane that specifically activates the Notch1 receptor and the Notch4 receptor as ligands.
  • DLL4 cytokines are currently reported in the field of neoplastic diseases, angiogenesis-related diseases, and cell proliferation and apoptosis research.
  • amino acid sequence of the DLL4 cytokine is reasonably modified in the art, and all belong to the scope of protection of the technical scheme of the present invention.
  • the present invention is directed to the defects of the prior art, and provides a new use of the DLL4 cytokine, the application of the DLL4 cytokine in the preparation of a medicament for treating fulminant hepatic failure.
  • the present invention is achieved by the following technical solutions:
  • the invention discloses the application of a DLL4 cytokine in the preparation of a medicament for treating fulminant hepatic failure.
  • the agents of the invention include DLL4 cytokines in pharmaceutically acceptable excipients, antioxidants, and carriers.
  • the medicament of the present invention is in the form of an injection or a lyophilized powder injection or suspension.
  • the DLL4 cytokine-containing agent of the present invention has a transport and storage temperature of 2 ° C to 8 ° C and a humidity of 45% to 75%.
  • the DLL4 cytokine-containing agent of the present invention is applied at a temperature of 15 ° C to 30 ° C and a humidity of 45% to 75%.
  • DLL4 cytokines in the preparation of agents for the treatment of liver failure.
  • DLL4 cytokines in the preparation of medicaments for the treatment of subacute liver failure.
  • DLL4 cytokines in the preparation of medicaments for the treatment of chronic liver failure.
  • DLL4 cytokines in the preparation of a medicament for the treatment of chronic acute liver failure.
  • the invention studies from biochemical indicators, immunohistochemistry, gene expression level, proteomics and the like, and finds that DLL4 cytokines can be used for the preparation of an effective treatment for fulminant hepatic failure, and the DLL4 cytokines can be combined with different solvents and Stabilizers produce drugs in different dosage forms.
  • the modes of administration that can be selected during use are intravenous and intramuscular.
  • DLL4 cytokine can overcome this problem.
  • DLL4 cytokines play an important role in the repair of the liver through activation of the DLL4-Notch pathway and can interact with other cytokines of the Notch pathway to promote the regeneration of the bile duct system.
  • DLL4 cytokines promote liver vascular endothelial cell regeneration, promote new angiogenesis, and improve blood circulation in diseased liver. Regulates the patient's immune function and reduces immune damage.
  • DLL4 cytokines can rapidly change the body's immune response in individuals with liver failure, rapidly inhibit inflammation, regulate fibroblast proliferation, inhibit the synthesis of fibrin, significantly reduce the degree of fibrosis in liver repair and avoid cirrhosis. .
  • DLL4 in the preparation of agents for the treatment of fulminant hepatic failure, it can significantly improve the biochemical indicators of patients, reduce bilirubin levels, reduce transaminase, improve blood coagulation, increase albumin levels, and promote hepatocytes and biliary cells. Health, prevent fatal complications such as massive gastrointestinal bleeding, severe hepatic encephalopathy and hepatorenal syndrome, significantly prolong the survival time of patients and improve survival.
  • DLL4 can significantly improve survival, DLL4 can significantly improve the prognosis and improve survival in patients with severe liver disease or end-stage liver disease with similar disease basis, disease progression and prognosis and fulminant hepatic failure. rate.
  • Figure 1 is a schematic diagram showing the survival time curve of the experimental group and the control group
  • Figure 2 is a schematic diagram showing the survival time curve of the experimental group and the control group
  • Figure 3 is a schematic diagram showing the survival time curves of rabbits in the experimental group and the control group;
  • Figure 4 is a graph showing the survival time curves of the experimental group and the control group.
  • Figure 5 is a schematic diagram showing the survival time curve of the experimental group and the control group of the subacute liver failure
  • Figure 6 is a schematic diagram showing the survival time curves of rabbits in the experimental group and the control group of chronic liver failure;
  • Fig. 7 is a graph showing the survival time curve of the experimental group and the control group of the chronic acute liver failure group.
  • the invention discloses the application of a DLL4 cytokine in the preparation of a medicament for treating fulminant hepatic failure, wherein the medicament forms are various, and the technical scheme of the invention is further illustrated by specific embodiments below:
  • Tween-80 was added to a 50% full volume of 0.9% NaCl sterilized aqueous solution.
  • DLL4 cytokines and proppant 1:5 ratio is completely dissolved in sterile water.
  • the pH of the 2 step solution is between 5.0 and 7.0.
  • the 3 step solution is placed in a lyophilizer and lyophilized. It was frozen at -40 ° C for 6 hours, vacuum freeze-dried at -40-0 ° C for 24 hours, and dried at 20 ° C for 2 hours to prepare a lyophilized powder.
  • the invention proves through a plurality of different animal model tests that the use of an agent containing DLL4 cytokines can significantly reduce liver lesions, improve biochemical indicators, prolong survival time, and improve survival rate in the treatment of fulminant hepatic failure. Therefore, the use of an agent containing a DLL4 cytokine for the clinical treatment of fulminant hepatic failure has broad development prospects and becomes a new method for treating severe liver diseases.
  • Example 1 Injection of DLL4 cytokine to treat large animal (young pig) model of fulminant hepatic failure
  • Animal models 30 male Chinese experimental piglets (8-10 kg) were randomly divided into two groups of 15 animals each. Each young pig was injected with D-gal 1.5g/kg into the jugular vein, resulting in a model of fulminant hepatic failure.
  • Test group DLL4 injection was intravenously administered at a fixed time point after D-gal injection at a dose of 10 ml/kg twice a day.
  • Control group An equal amount of physiological saline containing no DLL4 was injected.
  • Figure 1 is a schematic diagram showing the survival time curves of the experimental group and the control group, showing the survival rate of the experimental group and the control group.
  • the results showed that the 3-day survival rate of the pigs treated with the DLL4 cytokine was 93%, the 7-day survival rate was 85%, the 14-day survival rate was 85%, and the long-term stable survival was achieved.
  • the control group died in 72, and the average survival time was 46 hours. It can be seen that compared with the control group, the treatment of fulminant hepatic failure with the agent containing DLL4 cytokines can significantly prolong the survival time and improve the survival rate of the young pigs. It is indicated that the invention has significant advantages.
  • Example 2 Freeze-dried powder injection containing DLL4 cytokines for small animal (rat) model of fulminant hepatic failure
  • Animal model 100 male rats (200-250 g) were randomly divided into two groups of 50 each. Each rat was intraperitoneally injected with D-gal 1.5 g/kg to prepare a model of fulminant hepatic failure. The DLL4 lyophilized powder is mixed with water for injection into a suspension.
  • Test group 4 ml of DLL4 lyophilized powder suspension was intraperitoneally injected at a fixed time point after D-gal injection, twice a day.
  • Control group An equal amount of physiological saline containing no DLL4 was injected.
  • Fig. 2 is a graph showing the survival time curve of the experimental group and the control group, and Fig. 2 shows the survival rate of the experimental group and the control group: the results showed that the 3-day survival rate of the DLL4 treatment group was 96%, and the 7-day survival rate was obtained. 94%, 14-day survival rate was 94%, while the control group had a survival rate of 24% at 14 days.
  • the figure shows that compared with the control group, the treatment of fulminant hepatic failure with the agent containing DLL4 cytokines can significantly prolong the survival time of rats and improve the survival rate. It is indicated that the invention has significant advantages.
  • Example 3 Rabbit model containing DLL4 cytokine suspension for treatment of fulminant hepatic failure
  • Animal models 40 adult male rabbits (2000-2500 g) were randomly divided into two groups of 20 animals each. Each rabbit was intramuscularly injected with D-gal 1.5 g/kg to prepare a model of fulminant hepatic failure.
  • Test group 20 ml of DLL4 suspension was intramuscularly injected at a fixed time point after D-gal injection, twice a day.
  • Control group An equal amount of physiological saline containing no DLL4 was injected.
  • Figure 3 is a graph showing the survival time curves of rabbits in the experimental group and the control group, and Figure 3 shows the survival rate of the rabbits in the experimental group and the control group: the results showed that the 3-day survival rate of the rabbits in the DLL4 treatment group was 95%, and the survival rate was 7 days. 90%, 14-day survival rate was 90%, while the control group had a survival rate of 10% at 14 days.
  • the figure shows that compared with the control group, the treatment of fulminant hepatic failure with the agent containing DLL4 cytokines can significantly prolong the survival time of rabbits and improve the survival rate. It is indicated that the invention has significant advantages.
  • Example 4 A mouse model of fulminant hepatic failure treated with a suspension of DLL4 cytokines
  • Animal model 100 adult male mice (20-25 g) were randomly divided into two groups of 50 each. Each mouse was intramuscularly injected with CCl 4 to make a model of fulminant hepatic failure.
  • Test group 1 ml of DLL4 suspension was intraperitoneally injected at a fixed time point after CCl 4 injection, twice a day.
  • Control group An equal amount of physiological saline containing no DLL4 was injected.
  • Figure 4 is a graph showing the survival time curves of the experimental group and the control group, and Figure 4 shows the survival rate of the experimental group and the control group: the results showed that the 3-day survival rate of the mice in the DLL4 treatment group was 98%, and the 7-day survival rate was obtained. 94%, 14-day survival rate was 92%, while the control group had a survival rate of 30% at 14 days.
  • the figure shows that compared with the control group, the treatment of fulminant hepatic failure with the agent containing DLL4 cytokines can significantly prolong the survival time of mice and improve the survival rate. It is indicated that the invention has significant advantages.
  • Example 5 Freeze-dried powder injection containing DLL4 cytokines for treatment of subacute liver failure in small animal (rat) model
  • Animal model 100 male rats (200-250 g) were randomly divided into two groups of 50 each. Each rat was intraperitoneally injected with D-gal 1.5g/kg to prepare a subacute liver failure model. The DLL4 lyophilized powder is mixed with water for injection into a suspension.
  • Test group 4 ml of DLL4 lyophilized powder suspension was intraperitoneally injected at a fixed time point after D-gal injection, twice a day.
  • Control group An equal amount of physiological saline containing no DLL4 was injected.
  • Fig. 5 is a graph showing the survival time curve of the experimental group and the control group
  • Fig. 2 shows the survival rate of the experimental group and the control group: the results showed that the 3-day survival rate of the DLL4 treatment group was 98%, and the 7-day survival rate was obtained. 90%, 14-day survival rate was 90%, while the control group had a survival rate of 20% at 14 days.
  • the figure shows that compared to the control group, Treatment of subacute liver failure with agents containing DLL4 cytokines can significantly prolong survival and improve survival. It is indicated that the invention has significant advantages.
  • Example 6 Rabbit model containing chronic DLL failure treated with a suspension of DLL4 cytokines
  • Animal models 40 adult male rabbits (2000-2500 g) were randomly divided into two groups of 20 animals each. Each rabbit was intraperitoneally injected with a small dose of CCl 4 twice a week for 8 weeks to prepare a chronic liver failure model.
  • mice with chronic liver failure who were successfully modeled were injected intramuscularly with 5 ml of DLL4 cytokine suspension twice a day.
  • Control group An equal amount of physiological saline containing no DLL4 was injected.
  • Figure 6 is a graph showing the survival time curves of rabbits in the experimental group and the control group
  • Figure 3 shows the survival rate of the rabbits in the experimental group and the control group: the results showed that the 30-day survival rate of the rabbits in the DLL4 treatment group was 95%, and the survival rate was 90 days. 90%, while the control group had a survival rate of 40% at 30 days and a survival rate of 20% at 90 days.
  • the figure shows that compared with the control group, the treatment of chronic liver failure with the agent containing DLL4 cytokines can significantly prolong the survival time of rabbits and improve the survival rate. It is indicated that the invention has significant advantages.
  • Example 7 Suspension of DLL4 cytokines in a mouse model of chronic acute liver failure
  • Animal model 100 adult male mice (20-25 g) were randomly divided into two groups of 50 each. Each mouse was intraperitoneally injected with a small dose of CCl 4 to prepare a model of liver cirrhosis, and then cirrhotic mice were intraperitoneally injected with D-gal to prepare a model of chronic acute liver failure.
  • Test group 1 ml of DLL4 suspension was intraperitoneally injected at a fixed time point after D-gal injection, twice a day.
  • Control group An equal amount of physiological saline containing no DLL4 was injected.
  • Figure 7 is a graph showing the survival time curves of the experimental group and the control group
  • Figure 4 shows the survival rate of the experimental group and the control group: the results showed that the 3-day survival rate of the mice in the DLL4 treatment group was 98%, and the survival rate was 7 days. 94%, 14-day survival rate was 88%, while the control group had a survival rate of 25% at 14 days.
  • the figure shows that compared with the control group, the treatment of chronic acute liver failure with the agent containing DLL4 cytokines can significantly prolong the survival time of mice and improve the survival rate. It is indicated that the invention has significant advantages.

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Abstract

本发明提供了DLL4细胞因子在制备治疗暴发性肝功能衰竭药剂中的应用，除在暴发性肝功能衰竭治疗中，DLL4可显著提高生存率，在疾病基础、病情进展及预后与暴发性肝功能衰竭相似的严重肝病或终末期肝病中，DLL4也可明显改善预后，提高生存率。

Description

DLL4细胞因子在制备治疗暴发性肝功能衰竭药剂中的应用 技术领域
本发明属于临床医学、分子医学与再生医学领域，具体地说，是一种DLL4细胞因子系列药物治疗肝功能衰竭的新技术。
背景技术
暴发性肝功能衰竭是一类各种原因引起的肝脏大面积坏死所致的疾病，病情凶险，患者多于短期内死亡，常规内科综合治疗病死率高达80％以上。除原位肝移植外，目前尚缺乏特异性治疗方法。但由于严重的供肝短缺，大量患者在等待肝移植的过程中死去。如能找到一种或多种既可抑制肝脏坏死，又能迅速诱导宿主肝脏组织再生并长期发挥功能，并且能避免潜在副作用的细胞因子，对各种原因所致的暴发性肝功能衰竭治疗具有极其重要的意义。DLL4细胞因子是一种位于细胞膜上的I型跨膜蛋白，作为配体特异性活化Notch1受体和Notch4受体。
DLL4细胞因子目前于肿瘤性疾病、血管生成相关性疾病和细胞增殖及凋亡研究领域中报道较多。同时，DLL4细胞因子的氨基酸序列在本领域中的合理变形，均属于本发明技术方案要保护的范围。
发明内容
本发明正是针对现有技术存在的缺陷，提供了DLL4细胞因子的新用途—DLL4细胞因子在制备治疗暴发性肝功能衰竭药剂中的应用，本发明是通过以下技术方案来实现的：
本发明公开了一种DLL4细胞因子在制备治疗暴发性肝功能衰竭的药剂中的应用。
作为进一步地改进,本发明所述的药剂包括DLL4细胞因子在药学上可接受的赋形剂、抗氧化剂和载体。
作为进一步地改进，本发明所述的药剂的形式是注射液或冻干粉针剂或混悬液。
作为进一步地改进，本发明所述的含DLL4细胞因子的药剂的运输和贮存温度是2℃-8℃，湿度是45％-75％。
作为进一步地改进，本发明所述的含DLL4细胞因子的药剂的应用温度是15℃-30℃，湿度是45％-75％。
DLL4细胞因子在制备治疗肝衰竭药剂的应用。
DLL4细胞因子在制备治疗亚急性肝衰竭药剂中的应用。
DLL4细胞因子在制备治疗慢性肝衰竭药剂中的应用。
DLL4细胞因子在制备治疗慢加急性肝衰竭药剂中的应用。
与现有技术相比，本发明的有益效果如下：
本发明从生化指标、免疫组织化学，基因表达水平，蛋白组学等多个方面进行研究，发现DLL4细胞因子能够制备有效治疗暴发性肝功能衰竭药物中应用，可将DLL4细胞因子配合不同溶剂和稳定剂生产不同剂型剂量的药物。使用过程中可选择的给药方式有静脉注射和肌肉注射。
暴发性肝功能衰竭病程中，虽然有肝细胞再生，但胆管系统破坏后，新生的肝细胞难以存活并稳定的发挥作用。DLL4细胞因子可克服这一难题。DLL4细胞因子可以通过DLL4-Notch通路的活化作用在肝脏的修复过程中发挥重要作用，并可与Notch通路的其他细胞因子共同作用，促进胆管系统的再生。DLL4细胞因子可促进肝脏血管内皮细胞再生，促进新的血管生成，改善病变肝脏的血液循环。调节患者免疫功能，减轻免疫损伤。减轻肝细胞的变性坏死，抑制肝细胞凋亡，抑制肝脏大面积坏死。DLL4细胞因子可迅速改变肝功能衰竭个体的机体免疫反应，快速抑制炎症，调节成纤维细胞增生，抑制纤维蛋白的大量合成，可明显减轻肝脏修复过重中的纤维化程度，避免肝硬化的发生。DLL4在制备治疗暴发性肝功能衰竭的药剂的应用中，可显著改善患者的生化指标，降低胆红素水平，降低转氨酶，改善凝血功能，提高白蛋白水平，促进肝细胞和胆管细胞再 生，防止致死性并发症如上消化道大出血，严重肝性脑病和肝肾综合征的发生，显著延长患者存活时间，提高存活率。
除在暴发性肝功能衰竭治疗中，DLL4可显著提高生存率，在疾病基础、病情进展及预后与暴发性肝功能衰竭相似的严重肝病或终末期肝病中，DLL4也可明显改善预后，提高生存率。
附图说明
图1为实验组和对照组幼猪生存时间曲线示意图；
图2为实验组和对照组大鼠生存时间曲线示意图；
图3为实验组和对照组家兔生存时间曲线示意图；
图4为实验组和对照组小鼠生存时间曲线示意图。
图5为亚急性肝衰竭实验组和对照组大鼠生存时间曲线示意图；
图6为慢性肝衰竭实验组和对照组家兔生存时间曲线示意图；
图7为慢加急性肝衰竭实验组和对照组小鼠生存时间曲线示意图。
具体实施方式
本发明公开了一种DLL4细胞因子在制备治疗暴发性肝功能衰竭的药剂中的应用，其中的药剂形式各样，下面通过具体实施例子来对本发明的技术方案作进一步地说明：
一、含DLL4细胞因子的注射液
1、配置0.9％的NaCl灭菌水溶液1L。(1L规格)
2、0.009mg Tween-80加入50％全量体积的0.9％的NaCl灭菌水溶液中。
3、80ml的琥珀酸明胶加入2步骤所配置的溶液中。
4、处方剂量的DLL4细胞因子加入到3步骤所配置的溶液中。
5、用0.1mol/L的HCl调节pH值在5.0-7.0之间。
6、0.9％的NaCl灭菌水溶液定容至全量。
7、将制备的DLL4注射液分装保存(运输和贮存温度是2℃-8℃，湿度是45％-75％，优选运输和贮存温度是4℃；使用时15℃-30℃，湿度是45％-75％)。
二、含DLL4细胞因子的冻干粉针剂
1、制备足量的灭菌水。
2、DLL4细胞因子与支撑剂(甘露醇、山梨醇、右旋糖苷等)1:5比例完全溶解于灭菌水。
3、0.1mol/L的HCl调节2步骤溶液pH值在5.0-7.0之间。
4、3步骤溶液放入冻干机，冷冻干燥。-40℃冷冻6小时，-40-0℃真空冷冻干燥24小时，20℃干燥2小时制成冻干粉。
5、将制备的DLL4冻干粉分装保存(运输和贮存温度是2℃-8℃，湿度是45％-75％，优选运输和贮存温度是4℃；使用时15℃-30℃，湿度是45％-75％)。
三、含DLL4细胞因子混悬液(1000ml)
1、100克DLL4细胞因子粉碎成5-10微米大小的颗粒。
2、30g聚乙烯吡咯烷酮K-30(PvP)溶解于1000ml灭菌水中，制备溶液。
3、100g粉碎的DLL4细胞因子微粒加入步骤2所制备的溶液中。
4、使用稀盐酸溶液和稀氢氧化钠溶液调节pH值6.0-7.0之间。
5、在液体快速混合器中以3500转/分混合1小时。
6、将制备好的混悬液分装保存(运输和贮存温度是2℃-8℃，湿度是45％-75％，优选运输和贮存温度是4℃；使用时15℃-30℃，湿度是45％-75％)。
下面根据附图，通过具体实施例及对比例，对本发明的技术方案作进一步地说明：
本发明通过多种不同动物模型试验证明，使用含DLL4细胞因子的药剂在治疗暴发性肝衰竭可明显减轻肝脏病变程度，改善生化指标，延长存活时间，提高存活率。因此，将含DLL4细胞因子的药剂用于暴发性肝功能衰竭的临床治疗的药剂具有广阔的开发前景，成为治疗严重肝脏疾病的新方法。
实施例1：含DLL4细胞因子的注射液治疗暴发性肝衰竭的大动物(幼猪)模型
动物模型：30只雄性中国实验用幼猪(8-10公斤)随机分成两组，每组15只。每只幼猪颈静脉注射D-gal 1.5g/kg，造成暴发性肝功能衰竭模型。
试验组：分别在D-gal注射后的固定时间点多次静脉注射DLL4注射液，剂量：10ml/kg，一天两次。
对照组：注射等量不含DLL4的生理盐水。
对照组和实验组均未接受其他药物治。
图1为实验组和对照组幼猪生存时间曲线示意图，表示了实验组和对照组幼猪的生存率。结果显示，含DLL4细胞因子的药剂治疗组幼猪3天存活率93％，7天存活率85％，14天存活率85％并达到长期稳定存活。而对照组于72内全部死亡，平均生存时间46小时，由此可见，相比对照组，利用含DLL4细胞因子的药剂治疗暴发性肝衰竭可显著延长幼猪的生存时间，提高生存率。说明本发明具有明显优势。
实施例2：含DLL4细胞因子的冻干粉针剂治疗暴发性肝衰竭的小动物(大鼠)模型
动物模型：100只雄性大鼠(200-250克)随机分成两组，每组50只。每只大鼠腹腔注射D-gal 1.5g/kg，制成暴发性肝功能衰竭模型。将DLL4冻干粉与注射用水配置成混悬液。
试验组：分别在D-gal注射后的固定时间点予以腹腔注射DLL4冻干粉混悬液4ml，一天两次。
对照组：注射等量不含DLL4的生理盐水。
对照组和实验组均未接受其他药物治疗。
图2为实验组和对照组大鼠生存时间曲线示意图，图2表示了实验组和对照组大鼠的生存率：结果显示，DLL4治疗组的大鼠3天存活率96％，7天存活率94％，14天存活率94％，而对照组于14天存活率24％。由图表明，相比对照组，利用含DLL4细胞因子的药剂治疗暴发性肝衰竭可显著延长大鼠的生存时间，提高生存率。说明本发明具有明显优势。
实施例3：含DLL4细胞因子的混悬液治疗暴发性肝功能衰竭的家兔模型
动物模型：40只成年雄性家兔(2000-2500克)随机分成两组，每组20只。每只家兔肌肉注射D-gal 1.5g/kg，制成暴发性肝功能衰竭模型。
试验组：分别在D-gal注射后的固定时间点予以肌肉注射DLL4混悬液20ml，一天两次。
对照组：注射等量不含DLL4的生理盐水。
对照组和实验组均未接受其他药物治疗。
图3为实验组和对照组家兔生存时间曲线示意图，图3表示了实验组和对照组家兔的生存率：结果显示，DLL4治疗组的家兔3天存活率95％，7天存活率90％，14天存活率90％，而对照组于14天存活率10％。由图表明，相比对照组，利用含DLL4细胞因子的药剂治疗暴发性肝衰竭可显著延长家兔的生存时间，提高生存率。说明本发明具有明显优势。
实施例4：含DLL4细胞因子的混悬液治疗暴发性肝功能衰竭的小鼠模型
动物模型：100只成年雄性小鼠(20-25克)随机分成两组，每组50只。每只小鼠肌肉注射CCl4，制成暴发性肝功能衰竭模型。
试验组：分别在CCl4注射后的固定时间点予以腹腔注射DLL4混悬液1ml，一天两次。
对照组：注射等量不含DLL4的生理盐水。
对照组和实验组均未接受其他药物治疗。
图4为实验组和对照组小鼠生存时间曲线示意图，图4表示了实验组和对照组小鼠的生存率：结果显示，DLL4治疗组的小鼠3天存活率98％，7天存活率94％，14天存活率92％，而对照组于14天存活率30％。由图表明，相比对照组，利用含DLL4细胞因子的药剂治疗暴发性肝衰竭可显著延长小鼠的生存时间，提高生存率。说明本发明具有明显优势。
实施例5：含DLL4细胞因子的冻干粉针剂治疗亚急性肝衰竭的小动物(大鼠)模型
动物模型：100只雄性大鼠(200-250克)随机分成两组，每组50只。每只大鼠腹腔注射D-gal 1.5g/kg，制成亚急性肝功能衰竭模型。将DLL4冻干粉与注射用水配置成混悬液。
试验组：分别在D-gal注射后的固定时间点予以腹腔注射DLL4冻干粉混悬液4ml，一天两次。
对照组：注射等量不含DLL4的生理盐水。
对照组和实验组均未接受其他药物治疗。
图5为实验组和对照组大鼠生存时间曲线示意图，图2表示了实验组和对照组大鼠的生存率：结果显示，DLL4治疗组的大鼠3天存活率98％，7天存活率90％，14天存活率90％，而对照组于14天存活率20％。由图表明，相比对照组， 利用含DLL4细胞因子的药剂治疗亚急性肝衰竭可显著延长大鼠的生存时间，提高生存率。说明本发明具有明显优势。
实施例6：含DLL4细胞因子的混悬液治疗慢性肝功能衰竭的家兔模型
动物模型：40只成年雄性家兔(2000-2500克)随机分成两组，每组20只。每只家兔每周腹腔注射小剂量CCl4两次，共8周，制成慢性肝功能衰竭模型。
试验组：给予造模成功的慢性肝功能衰竭家兔DLL4细胞因子混悬液肌肉注射5ml，一天两次。
对照组：注射等量不含DLL4的生理盐水。
对照组和实验组均未接受其他药物治疗。
图6为实验组和对照组家兔生存时间曲线示意图，图3表示了实验组和对照组家兔的生存率：结果显示，DLL4治疗组的家兔30天存活率95％，90天存活率90％，而对照组于30天存活率40％，90天存活率20％。由图表明，相比对照组，利用含DLL4细胞因子的药剂治疗慢性肝衰竭可显著延长家兔的生存时间，提高生存率。说明本发明具有明显优势。
实施例7：含DLL4细胞因子的混悬液治疗慢加急性肝功能衰竭的小鼠模型
动物模型：100只成年雄性小鼠(20-25克)随机分成两组，每组50只。每只小鼠腹腔注射小剂量CCl4，制成肝硬化模型，然后给予肝硬化小鼠腹腔注射D-gal，制成慢加急性肝衰竭模型。
试验组：分别在D-gal注射后的固定时间点予以腹腔注射DLL4混悬液1ml，一天两次。
对照组：注射等量不含DLL4的生理盐水。
对照组和实验组均未接受其他药物治疗。
图7为实验组和对照组小鼠生存时间曲线示意图，图4表示了实验组和对照组小鼠的生存率：结果显示，DLL4治疗组的小鼠3天存活率98％，7天存活率94％，14天存活率88％，而对照组于14天存活率25％。由图表明，相比对照组，利用含DLL4细胞因子的药剂治疗慢加急性肝衰竭可显著延长小鼠的生存时间，提高生存率。说明本发明具有明显优势。
以上例举的仅是本发明的优选实施方式，本发明并不限于以上实施例，本领 域技术人员在不脱离本发明的精神和构思的前提下直接导出或联想到的其他改进和变化，均应认为包含在本发明的保护范围内。

Claims (8)

  1. DLL4细胞因子在制备治疗暴发性肝功能衰竭的药剂中的应用。
  2. 根据权利要求1所述的DLL4细胞因子在制备治疗暴发性肝功能衰竭的药剂中的应用，其特征在于，所述的药剂包括DLL4细胞因子在药学上可接受的赋形剂、抗氧化剂和载体。
  3. 根据权利要求2所述的DLL4细胞因子在制备治疗暴发性肝功能衰竭的药剂中的应用，其特征在于，所述的药剂的形式是注射液或冻干粉针剂或混悬液。
  4. 根据权利要求1或2或3所述的DLL4细胞因子在制备治疗暴发性肝功能衰竭的药剂中的应用，其特征在于，所述的含DLL4细胞因子的药剂的运输和贮存温度是2℃-8℃，湿度是45％-75％。
  5. DLL4细胞因子在制备治疗肝衰竭药剂的应用。
  6. DLL4细胞因子在制备治疗亚急性肝衰竭药剂中的应用。
  7. DLL4细胞因子在制备治疗慢性肝衰竭药剂中的应用。
  8. DLL4细胞因子在制备治疗慢加急性肝衰竭药剂中的应用。
PCT/CN2016/075574 2015-09-15 2016-03-04 Dll4细胞因子在制备治疗暴发性肝功能衰竭药剂中的应用 WO2017045377A1 (zh)

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