Methods of validating candidate compounds for use in treating copd and other diseases

Abstract

The present invention relates to methods of diagnosing, monitoring, and treating elastin fiber injuries. In additional preferred embodiments, the present invention relates to methods of validating candidate compounds for use in treating chronic obstructive pulmonary disease (COPD), chronic bronchitis, emphysema, refractory asthma, and other related diseases. Examples of such methods include determining if the candidate compound decreases the degradation of elastic fiber in a patient administered the candidate compound by measuring, using mass spectrometry employing an internal standard, a marker of elastic fiber degradation in a sample of a body fluid or a tissue of the patient. The invention provides that a decrease in the presence of the marker compared to a control validates that the candidate compound is effective to treat, prevent, or ameliorate the disease.

What is claimed is: 
     
         1 . A method of validating whether a candidate compound is effective to treat, prevent, or ameliorate the effects of a disease characterized by elastic fiber injury comprising determining if the candidate compound decreases the degradation of an elastic fiber in a patient administered the candidate compound by measuring, using mass spectrometry employing an internal standard, a marker of elastic fiber degradation in a sample of a body fluid or a tissue of the patient, wherein a decrease in the presence of the marker compared to a control validates that the candidate compound is effective to treat, prevent, or ameliorate the disease. 
     
     
         2 . The method according to  claim 1 , wherein the elastic fiber injury is elastin degradation. 
     
     
         3 . The method according to  claim 1 , wherein the disease is selected from the group consisting of chronic obstructive pulmonary disease (COPD), COPD with alpha-1 antitrypsin deficiency (AATD), chronic bronchitis, emphysema, and refractory asthma. 
     
     
         4 . The method according to  claim 1 , wherein the disease is COPD. 
     
     
         5 . The method according to  claim 1 , wherein the marker of elastic fiber degradation is selected from the group consisting of desmosine, isodesmosine, and combinations thereof. 
     
     
         6 . The method according to  claim 1 , wherein the marker is both desmosine and isodesmosine. 
     
     
         7 . The method according to  claim 1 , wherein the body fluid is selected from the group consisting of urine, plasma, and sputum. 
     
     
         8 . The method according to  claim 5 , wherein both desmosine and isodesmosine are measured in plasma. 
     
     
         9 . The method according to  claim 5 , wherein total free desmosine and isodesmosine are measured in urine. 
     
     
         10 . The method according to  claim 1 , wherein the candidate compound is selected from the group consisting of hyaluronic acid, polysaccharide, carbohydrate, small molecules, and RNAi. 
     
     
         11 . The method according to  claim 1 , wherein tandem mass spectrometry is used. 
     
     
         12 . The method according to  claim 1 , wherein the internal standard used in the mass spectrometry is acylated pyridinoline. 
     
     
         13 . The method according to  claim 1 , wherein tandem mass spectrometry is used and the internal standard is acylated pyridinoline. 
     
     
         14 . A method of validating whether a candidate compound is effective to treat, prevent, or ameliorate the effects of chronic obstructive pulmonary disease (COPD) comprising determining if the candidate compound decreases the degradation of elastin in a patient administered the candidate compound by measuring, using mass spectrometry employing an internal standard, the amount of desmosine and isodesmosine in a sample of a body fluid or tissue of the patient, wherein a decrease in the presence of desmosine or isodesmosine compared to a control validates that the candidate compound is effective to treat, prevent, or ameliorate the disease. 
     
     
         15 . The method according to  claim 14 , wherein the body fluid is selected from the group consisting of urine, plasma, and sputum. 
     
     
         16 . The method according to  claim 15 , wherein both desmosine and isodesmosine are measured in plasma. 
     
     
         17 . The method according to  claim 15 , wherein total free desmosine and isodesmosine are measured in urine. 
     
     
         18 . The method according to  claim 14 , wherein tandem mass spectrometry is used. 
     
     
         19 . The method according to  claim 14 , wherein the internal standard used in the mass spectrometry is acylated pyridinoline. 
     
     
         20 . The method according to  claim 14 , wherein tandem mass spectrometry is used and the internal standard is acylated pyridinoline. 
     
     
         21 . A method of validating whether a candidate compound is effective to treat, prevent, or ameliorate the effects of chronic obstructive pulmonary disease (COPD) comprising determining if the candidate compound decreases the degradation of elastin in a patient administered the candidate compound by measuring, using mass spectrometry employing an internal standard, the amount of desmosine and isodesmosine in a sample from the patient selected from the group consisting of plasma, urine, and sputum, wherein a decrease in the presence of desmosine and isodesmosine compared to a control validates that the candidate compound is effective to treat, prevent, or ameliorate the disease. 
     
     
         22 . The method according to  claim 21 , wherein tandem mass spectrometry is used. 
     
     
         23 . The method according to  claim 21 , wherein the internal standard used in the mass spectrometry is acylated pyridinoline. 
     
     
         24 . The method according to  claim 21 , wherein tandem mass spectrometry is used and the internal standard is acylated pyridinoline. 
     
     
         25 . A method for identifying candidate compounds that are effective to treat, prevent, or ameliorate the effects of a disease characterized by elastic fiber injury comprising:
 (a) administering a candidate compound to a cell culture model of the disease;   (b) measuring, by mass spectrometry using an internal standard, the amount of a marker of elastic fiber injury in the cell culture administered the candidate compound; and   (c) determining whether the amount of the marker produced by the cell culture administered the candidate compound is different compared to a control cell culture absent the candidate compound, wherein a decrease in the amount of the marker produced by the cell culture administered the candidate compound compared to the control cell culture identifies the candidate compound as effective to treat, prevent, or ameliorate the effects of the disease.   
     
     
         26 . The method according to  claim 25 , wherein the elastic fiber injury is elastin degradation. 
     
     
         27 . The method according to  claim 25 , wherein the disease is selected from the group consisting of chronic obstructive pulmonary disease (COPD), COPD with AATD, chronic bronchitis, emphysema, and refractory asthma. 
     
     
         28 . The method according to  claim 25 , wherein the disease is COPD. 
     
     
         29 . The method according to  claim 25 , wherein the marker is selected from the group consisting of desmosine, isodesmosine, and combinations thereof. 
     
     
         30 . The method according to  claim 25 , wherein both desmosine and isodesmosine are measured. 
     
     
         31 . The method according to  claim 25 , wherein the candidate compound is selected from the group consisting of hyaluronic acid, polysaccharide, carbohydrate, small molecules, and RNAi. 
     
     
         32 . The method according to  claim 25 , wherein tandem mass spectrometry is used. 
     
     
         33 . The method according to  claim 25 , wherein the internal standard used in the mass spectrometry is acylated pyridinoline. 
     
     
         34 . The method according to  claim 25 , wherein tandem mass spectrometry is used and the internal standard is acylated pyridinoline. 
     
     
         35 . A method for identifying candidate compounds that are effective to treat, prevent, or ameliorate the effects of a disease characterized by elastin degradation comprising:
 (a) administering a candidate compound to a cell culture model of the disease;   (b) measuring, by mass spectrometry using an internal standard, the amount of desmosine and isodesmosine in the cell culture administered the candidate compound; and   (c) determining whether the amount of desmosine and isodesmosine produced by the cell culture administered the candidate compound is different compared to a control cell culture absent the candidate compound, wherein a decrease in the amount of the desmosine and isodesmosine produced by the cell culture administered the candidate compound compared to the control cell culture identifies the candidate compound as effective to treat, prevent, or ameliorate the effects of the disease.   
     
     
         36 . The method according to  claim 35 , wherein tandem mass spectrometry is used. 
     
     
         37 . The method according to  claim 35 , wherein the internal standard used in the mass spectrometry is acylated pyridinoline. 
     
     
         38 . The method according to  claim 35 , wherein tandem mass spectrometry is used and the internal standard is acylated pyridinoline.