Use of long-acting human recombinant soluble tumor necrosis factor a receptor in the preparation of drugs for preventing and treating severe liver injury on chronic liver disease

Abstract

The proposed invention involves the use of new drugs with the recombinant soluble tumor necrosis α receptor (HusTNFR) and belongs to the gene engineering technology and gene function application field. This invention uses type I or type II long-acting HusTNFR (LHusTNFR) to perform an intervention for severe liver injury in rats with chronic liver disease using 5 types of animal models. The results showed that LHusTNFR, which has a half-life of 12-140 hours, shows excellent efficacy for preventing the development of severe liver injury on chronic liver disease and for treating early-stage severe liver injury on chronic liver disease. It also significantly reduced the mortality of the model animals. Its efficacy for the prevention and treatment of early-stage severe liver injury on chronic liver disease was significantly better than that of non-LHusTNFR.

1 - 13 . (canceled) 
     
     
         14 . The use of long-acting soluble tumor necrosis factor α receptor (LHusTNFR) in the preparation of drugs to prevent and/or treat severe liver injury on chronic liver disease or hepatic necrosis on the basis of chronic liver diseases.
 The mentioned chronic liver disease with severe liver injury (also called severe liver injury on chronic liver disease) refers to diseases that occur in the presence of chronic liver disease and are caused by viruses, immune injury, toxins (or drugs), alcohol, or a high-fat diet; severe liver injury also refers to a class of liver diseases that result from the rapid deterioration of liver functions caused by a recently suffered major injury in the liver that induces acute and massive hepatocyte necrosis that can develop into multiple organ failure with a short-term mortality rate greater than 20%. 
 The mentioned hepatocyte necrosis refers to acute and large area hepatocyte necrosis (massive/submassive hepatic necrosis), which is one of the characteristic manifestations of liver pathology. 
 
     
     
         15 . The use of  claim 14 , wherein the mentioned treatment is the treatment of early-stage chronic liver diseases with severe liver injury and early-stage hepatocyte necrosis related diseases on the basis of chronic liver diseases. 
     
     
         16 . The use of  claim 15 , wherein the said early-stage liver diseases with severe liver injury refer to the presence of systemic inflammatory response syndrome (SIRS) in chronic liver diseases with severe liver injury. 
     
     
         17 . The use of  claim 16 , wherein the mentioned SIRS refers to the increase of peripheral blood pro-inflammatory cytokines TNFα and IL-6 or the increase of both pro-inflammatory cytokines TNFα and IL-6 and the anti-inflammatory cytokine IL-10. 
     
     
         18 . The use as stated in  claim 14 , wherein the mentioned long-acting HusTNFR has a half-life of 12-140 hours. 
     
     
         19 . The use as mentioned in  claim 14 , in which the said long-acting HusTNFR is selected from the following:
 a. a fusion protein between human type I tumor necrosis factor α receptor and the human IgG1:Fc fragment,   b. a fusion protein between human type II tumor necrosis factor α receptor and the human IgG1:Fc fragment,   c. a conjugated product between the amino terminus of human type I tumor necrosis factor α receptor and PEG,   d. a conjugated product between the carboxyl terminus of human type I tumor necrosis factor α receptor and PEG,   e. a conjugated protein between the amino terminus of human type II tumor necrosis factor α receptor and PEG,   f. a conjugated product between the carboxyl terminus of human type II tumor necrosis factor α receptor and PEG,   g. a human type I tumor necrosis factor α receptor protein product embedded in a PEG-liposome mixture, or   h. a human type II tumor necrosis factor α receptor protein product embedded in a PEG-liposome mixture.   
     
     
         20 . The use as mentioned in  claim 14 , wherein the said long-acting HusTNFR can significantly reduce the death rate of animals with chronic liver diseases with severe liver injury;
 Prevent the massive/submassive necrosis of liver tissues;   Decrease caspase 3 activity levels by 70-80%;   Decrease the TUNEL-positive cell count/high power field in liver pathology by more than 80%;   Decrease the liver tissue injury score by 40-70%;   Essentially eliminate the pathological characteristics of massive/submassive hepatic necrosis;   Decrease nuclear factor κB (NF-κB) levels in the liver by 30-50%;   Decrease the peak values of TNFα, IL-6, and IL-10 in the peripheral blood by 40-95%;   Decrease the peak values of TNFα and IL-6 in the liver by 40-80%; and/or   Increase the IL-22 and IL-22 receptors in the liver by 2- to 5-fold.   
     
     
         21 . The use as stated in  claim 15 , wherein the said LHusTNFR can significantly reduce the mortality rate of animals with early-stage chronic liver disease with severe liver injury;
 Prevent the massive/submassive necrosis of liver tissues;   Decrease caspase 3 activity levels by 70-80%;   Decrease the TUNEL-positive cell count/high power field in liver pathology by more than 80%;   Decrease the liver tissue injury score by 40-70%;   Essentially eliminate the pathological characteristics of massive/submassive hepatic necrosis;   Decrease NF-κB levels in the liver by 30-50%;   Decrease the peak values of TNFα, IL-6, and IL-10 in the peripheral blood by 40-95%;   Decrease the peak values of TNFα and IL-6 in the liver by 40-80%; and/or   Increase the IL-22 and IL-22 receptors in the liver by 2- to 5-fold.   
     
     
         22 . The use as stated in  claim 14 , wherein the said severe liver injury on chronic liver disease includes the following: severe alcoholic hepatitis, hepatitis virus associated hepatitis, acute-on-chronic liver failure caused by liver fibrosis or cirrhosis, severe liver injury caused by non-alcoholic fatty liver disease, severe liver injury caused by autoimmune liver disease, and drug-induced severe liver injury caused by chronic liver disease. 
     
     
         23 . A pharmaceutical composition, wherein the pharmaceutical composition comprises the following:
 (i) an effective amount of selected long-acting HusTNFR:   a. a fusion protein between human type I tumor necrosis factor α receptor and the human IgG1:Fc fragment,   b. a fusion protein between human type II tumor necrosis factor α receptor and the human IgG1:Fc fragment,   c. a conjugated product between the amino terminus of human type I tumor necrosis factor α receptor and PEG,   d. a conjugated product between the carboxyl terminus of human type I tumor necrosis factor α receptor and PEG,   e. a conjugated protein between the amino terminus of human type II tumor necrosis factor α receptor and PEG,   f. a conjugated product between the carboxyl terminus of human type II tumor necrosis factor α receptor and PEG,   h. a human type I tumor necrosis factor α receptor protein product embedded in a PEG-liposome mixture, or   i. a human type II tumor necrosis factor α receptor protein product embedded in a PEG-liposome mixture,   and   (ii) pharmacologically acceptable carriers.   The pharmacological composition also contains an effective amount of one or more drugs among the following:   (iii) α-thymosin, human serum albumin, polyene phosphatidylcholine, a variety of coenzyme vitamins, and Xuebijing (traditional Chinese medicine).   
     
     
         24 . A method for the prevention of severe liver injury on chronic liver disease and hepatocyte necrosis caused by chronic liver diseases wherein patients who require treatment are given LHusTNFR. 
     
     
         25 . A method for the treatment of early-stage severe liver injury on chronic liver disease and early-stage hepatocyte necrosis caused by chronic liver diseases wherein patients who require treatment are given LHusTNFR. 
     
     
         26 . The use of the methods as stated in  claim 24  in which the said LHusTNFR is selected from the following group:
 a. a fusion protein between human type I tumor necrosis factor α receptor and the human IgG1:Fc fragment, 
 b. a fusion protein between human type II tumor necrosis factor α receptor and the human IgG1:Fc fragment, 
 c. a conjugated product between the amino terminus of human type I tumor necrosis factor α receptor and PEG, 
 d. a conjugated product between the carboxyl terminus of human type I tumor necrosis factor α receptor and PEG, 
 e. a conjugated protein between the amino terminus of human type II tumor necrosis factor α receptor and PEG, 
 f. a conjugated product between the carboxyl terminus of human type II tumor necrosis factor α receptor and PEG, 
 h. a human type I tumor necrosis factor α receptor protein product embedded in a PEG-liposome mixture, or 
 i. a human type II tumor necrosis factor α receptor protein product embedded in a PEG-liposome mixture. 
 
     
     
         27 . The use of the methods as stated in  claim 25  in which the said LHusTNFR is selected from the following group:
 a. a fusion protein between human type I tumor necrosis factor α receptor and the human IgG1:Fc fragment, 
 b. a fusion protein between human type II tumor necrosis factor α receptor and the human IgG1:Fc fragment, 
 c. a conjugated product between the amino terminus of human type I tumor necrosis factor α receptor and PEG, 
 d. a conjugated product between the carboxyl terminus of human type I tumor necrosis factor α receptor and PEG, 
 e. a conjugated protein between the amino terminus of human type II tumor necrosis factor α receptor and PEG, 
 f. a conjugated product between the carboxyl terminus of human type II tumor necrosis factor α receptor and PEG, 
 h. a human type I tumor necrosis factor α receptor protein product embedded in a PEG-liposome mixture, or 
 i. a human type II tumor necrosis factor α receptor protein product embedded in a PEG-liposome mixture.