Agents, methods and kits for treating and detecting diabetic nephropathy and proliferative diseases and methods for inhibiting expression of type iv collagen

Abstract

A method of detecting proliferative diseases causing sclerosis, comprising measuring the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1, phosphorylated Smad1, activin receptor-like kinase 1, activin receptor-like kinase 3 and bone morphogenetic proteins in a biological sample. A kit therefor. A prophylactic and/or therapeutic agent for proliferative diseases causing sclerosis, comprising as an active ingredient a substance having an inhibitory effect on the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1. A method of identifying substances effective in preventing and/or treating proliferative diseases causing sclerosis, comprising judging whether or not a test substance inhibits the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1. A kit therefor.

1 . A method of detecting and treating diabetic nephropathy, which method comprises:
 contacting a biological sample from a subject with a pair of primers that amplify Smad1 and/or a substance having Smad1-activating effect;   detecting the presence or absence of expression of Smad1 and/or a substance having Smad1-activating effect in the biological sample from the subject;   identifying the subject as having diabetic nephropathy if the presence of the expression of Smad1 and/or the substance having Smad1-activating effect is detected;   administering to the subject a drug comprising a substance having an inhibitory effect on the expression of Smad1,   wherein the substance having Smad1-activating effect is selected from the group consisting of activin receptor-like kinase 1 (ALK1), activin receptor-like kinase 3 (ALK3) and bone morphogenetic protein BMP2.   
     
     
         2 . A method of evaluating the degree of progress and/or the efficacy of treatment of diabetic nephropathy, which method comprises:
 measuring the expression level of Smad1 and/or a substance having Smad1-activating effect in a first biological sample from a subject at a first time point by contacting the first biological sample with a pair of primers that amplify Smad1 and/or a substance having Smad1-activating effect;   administering to the subject a drug comprising a substance having an inhibitory effect on the expression of Smad1;   measuring the expression level of Smad1 and/or the substance having Smad1-activating effect in a second biological sample from the subject at a second time point by contacting the second biological sample with a pair of primers that amplify Smad1 and/or a substance having Smad1-activating effect;   comparing the expression level of Smad1 and/or a substance having Smad1-activating effect from the first and second biological sample,   wherein an increase in the expression level of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates progression of diabetic nephropathy, and   wherein a decrease in the expression of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates that the diabetic nephropathy has not progressed,   wherein the substance having Smad1-activating effect is selected from the group consisting of activin receptor-like kinase 1 (ALK1), activin receptor-like kinase 3 (ALK3) and bone morphogenetic protein BMP2, and   wherein an increase in the expression level of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates that the treatment of diabetic nephropathy is not effective, wherein a decrease in the expression level of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates that the treatment of diabetic nephropathy is effective.   
     
     
         3 . A kit for detecting diabetic nephropathy, comprising a reagent(s) for measuring the expression of Smad1 and/or a substance having Smad1-activating effect. 
     
     
         4 . A kit for evaluating the degree of progress and/or the efficacy of treatment of diabetic nephropathy, comprising a reagent(s) for measuring the expression of Smad1 and/or a substance having Smad1-activating effect. 
     
     
         5 . A prophylactic and/or therapeutic agent for proliferative diseases causing sclerosis, comprising as an active ingredient a substance having an inhibitory effect on the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1. 
     
     
         6 . A drug inhibiting the increase of extracellular matrix, comprising as an active ingredient a substance having an inhibitory effect on the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1. 
     
     
         7 . A drug inhibiting the expression of α1 type IV collagen, comprising as an active ingredient a substance having an inhibitory effect on the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1. 
     
     
         8 . A method for producing a prophylactic and/or therapeutic effect on proliferative diseases causing sclerosis, comprising administering to a subject a drug including a substance having an inhibitory effect on the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1. 
     
     
         9 . A method of inhibiting an increase of extracellular matrix, comprising providing to a sample a substance having an inhibitory effect on the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1. 
     
     
         10 . A method of inhibiting an expression of α1 type IV collagen, comprising administering to a subject a drug including a substance having an inhibitory effect on the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1. 
     
     
         11 . A method of inhibiting an expression of α1 type IV collagen, comprising providing to a sample a substance having an inhibitory effect on the expression of at least one substance selected from the group consisting of STAT3, phosphorylated STAT3, Smad1 and phosphorylated Smad1. 
     
     
         12 . A method of  claim 8 , wherein the substance to be administered includes antisense oligonucleotides to Smad1, Smad1 Antagonistic Effector, an anti-PDGF β receptor antibody, or antisense oligonucleotides to STAT3. 
     
     
         13 . A method of  claim 8 , wherein the substance to be administered has a sequence comprising SEQ ID NO: 13. 
     
     
         14 . The method according to  claim 2 , wherein the first time point is before treatment and the second time point is after treatment. 
     
     
         15 . The method according to  claim 1 , wherein the biological sample is selected from the group consisting of renal tissue sections, blood, sera, and urine. 
     
     
         16 . The method of  claim 2 , wherein the biological sample is selected from the group consisting of renal tissue sections, blood, sera and urine. 
     
     
         17 . The method according to  claim 1 , wherein detection of expression is determined at the nucleic acid level or the protein level. 
     
     
         18 . The method according to  claim 2 , wherein the level of expression is measured at the nucleic acid level or the protein level. 
     
     
         19 . The method according to  claim 1 , wherein the presence or absence of Smad1 expression is detected. 
     
     
         20 . The method according to  claim 2 , wherein the expression of Smad1 is measured. 
     
     
         21 . A method of evaluating the degree of progress and/or the efficacy of treatment of diabetic nephropathy and treating a subject, which method comprises:
 measuring the expression level of Smad1 and/or a substance having Smad1-activating effect in a first biological sample from a subject at a first time point;   administering to the subject a drug comprising a substance having an inhibitory effect on the expression of Smad1;   measuring the expression level of Smad1 and/or the substance having Smad1-activating effect in a second biological sample from the subject at a second time point;   comparing the expression level of Smad1 and/or a substance having Smad1-activating effect from the first and second biological sample,   wherein an increase in the expression level of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates progression of diabetic nephropathy, and   wherein a decrease in the expression of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates that the diabetic nephropathy has not progressed,   wherein the substance having Smad1-activating effect is selected from the group consisting of activin receptor-like kinase 1 (ALK1), activin receptor-like kinase 3 (ALK3) and bone morphogenetic protein BMP2, and   wherein an increase in the expression level of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates that the treatment of diabetic nephropathy is not effective, wherein a decrease in the expression level of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates that the treatment of diabetic nephropathy is effective.   
     
     
         22 . A method of detecting and treating diabetic nephropathy, which method comprises:
 detecting the presence of expression of Smad1 and/or a substance having Smad1-activating effect in a biological sample from a subject,   identifying the subject as having diabetic nephropathy if the presence of the expression of Smad1 and/or the substance having Smad1-activating effect is detected,   administering to the subject a drug comprising a substance having an inhibitory effect on the expression of Smad1,   wherein the substance having Smad1-activating effect is selected from the group consisting of activin receptor-like kinase 1 (ALK1), activin receptor-like kinase 3 (ALK3) and bone morphogenetic protein BMP2.   
     
     
         23 . The method according to  claim 21 , wherein the drug comprises an antisense oligonucleotide to Smad1. 
     
     
         24 . The method according to  claim 22 , wherein the drug comprises an antisense oligonucleotide to Smad1. 
     
     
         25 . A method of detecting diabetic nephropathy, which method comprises:
 contacting a biological sample from a subject with a pair of primers that amplify Smad1 and/or a substance having Smad1-activating effect;   detecting the presence or absence of expression of Smad1 and/or a substance having Smad1-activating effect in the biological sample from the subject,   identifying the subject as having diabetic nephropathy if the presence of the expression of Smad1 and/or the substance having Smad1-activating effect is detected,   wherein the substance having Smad1-activating effect is selected from the group consisting of activin receptor-like kinase 1 (ALK1), activin receptor-like kinase 3 (ALK3) and bone morphogenetic protein BMP2,   wherein the pair of primers are selected from the group consisting of SEQ ID NO: 5 and SEQ ID NO: 6; SEQ ID NO: 7 and SEQ ID NO: 8; SEQ ID NO: 9 and SEQ ID NO: 10; SEQ ID NO: 11 and SEQ ID NO: 12; and SEQ ID NO: 23 and SEQ ID NO: 24.   
     
     
         26 . A method of evaluating the degree of progress and/or the efficacy of treatment of diabetic nephropathy, which method comprises:
 measuring the expression level of Smad1 and/or a substance having Smad1-activating effect in a first biological sample from a subject at a first time point by contacting the first biological sample with a pair of primers that amplify Smad1 and/or a substance having Smad1-activating effect;   measuring the expression level of Smad1 and/or the substance having Smad1-activating effect in a second biological sample from the subject at a second time point by contacting the second biological sample with a pair of primers that amplify Smad1 and/or a substance having Smad1-activating effect;   comparing the expression level of Smad1 and/or a substance having Smad1-activating effect from the first and second biological sample;   wherein an increase in the expression level of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates progression of diabetic nephropathy, and   wherein a decrease in the expression of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates that the diabetic nephropathy has not progressed,   wherein the substance having Smad1-activating effect is selected from the group consisting of activin receptor-like kinase 1 (ALK1), activin receptor-like kinase 3 (ALK3) and bone morphogenetic protein BMP2,   wherein an increase in the expression level of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates that the treatment of diabetic nephropathy is not effective, wherein a decrease in the expression level of Smad1 and/or a substance having Smad1-activating effect in the second biological sample in comparison to the first biological sample indicates that the treatment of diabetic nephropathy is effective, and   wherein the pair of primers are selected from the group consisting of SEQ ID NO: 5 and SEQ ID NO: 6; SEQ ID NO: 7 and SEQ ID NO: 8; SEQ ID NO: 9 and SEQ ID NO: 10; SEQ ID NO: 11 and SEQ ID NO: 12; and SEQ ID NO: 23 and SEQ ID NO: 24.