Methods and Means for Treatment of Osteoarthritis

Abstract

The invention relates to the field of medicinal research, cartilage physiology and diseases involving the degeneration of cartilage tissue. More specifically, the invention relates to methods and means for identifying compounds that inhibit catabolic processes in chondrocytes and that decrease the degradation of cartilage and/or ECM. The invention also relates to the compounds that are useful in the treatment of osteoarthritis. The invention also relates to targets, the modulation of which results in a decrease in the degradation of ECM and/or cartilage and decrease inflammation. In addition, the invention relates to compositions and methods for the use thereof in treating conditions that are characterized by the degradation of ECM and/or cartilage and inflammation.

1 . A method for identifying a compound that inhibits ECM and/or cartilage degradation, comprising:
 (a) contacting a compound with a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 39, 40, 27, 42, 22-26, 28-38, and 41 and fragments thereof; and   (b) measuring a compound-polypeptide property related to ECM and/or cartilage degradation.   
     
     
         2 . The method according to  claim 1 , wherein said polypeptide is in an in vitro cell-free preparation. 
     
     
         3 . The method according to  claim 1 , wherein said polypeptide is present in a mammalian cell. 
     
     
         4 . The method of  claim 2 , wherein said property is a binding affinity of said compound to said polypeptide. 
     
     
         5 . The method of  claim 1 , wherein the method is used to identify compounds that inhibit the catabolic process of chondrocytes. 
     
     
         6 . The method of  claim 4 , which additionally comprises the steps of:
 c) contacting a population of mammalian cells expressing said polypeptide with the compound that exhibits a binding affinity of at least 10 micromolar; and   d) identifying a compound that inhibits ECM and/or cartilage degradation.   
     
     
         7 . The method of  claim 1 , wherein said property is the expression and/or activity of MMP1, MMP3, MMP8, MMP13, MMP14, and/or ADAMTS4. 
     
     
         8 . The method of  claim 7 , wherein said property is the expression and/or activity of MMP13. 
     
     
         9 . The method according to  claim 1 , wherein said property is the activity of said polypeptide. 
     
     
         10 . The method according to  claim 1 , wherein said property is the expression of said polypeptide. 
     
     
         11 . The method according to  claim 9 , which additionally comprises the steps of:
 c) contacting a population of mammalian cells expressing said polypeptide with the compound that significantly inhibits the expression or activity of the polypeptide; and   d) identifying the compound that inhibits ECM and/or cartilage degradation.   
     
     
         12 . The method according to  claim 1 , which additionally comprises the step of comparing the compound to be tested to a control. 
     
     
         13 . The method according to  claim 12 , wherein said control is where the polypeptide has not been contacted with said compound. 
     
     
         14 . The method according to  claim 6 , which additionally comprises the step of comparing the compound to a control, wherein said control is a population of mammalian cells that does not express said polypeptide. 
     
     
         15 . The method according to  claim 1 , wherein said compound is selected from the group consisting of compounds of a commercially available screening library and compounds having binding affinity for a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 39, 40, 27, 42, 22-26, 28-38, and 41. 
     
     
         16 . The method according to  claim 1 , wherein said compound is a peptide in a phage display library or an antibody fragment library. 
     
     
         17 . An agent effective in inhibiting ECM and/or cartilage degradation, selected from the group consisting of an antisense polynucleotide, a ribozyme, and a small interfering RNA (siRNA), wherein said agent comprises a nucleic acid sequence complementary to, or engineered from, a naturally-occurring polynucleotide sequence of about 17 to about 30 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 18, 19, 6, 21, 1-5, 7-17, and 20. 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . (canceled) 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . A pharmaceutical composition comprising a therapeutically effective amount of an agent according to  claim 17  in admixture with a pharmaceutically acceptable carrier. 
     
     
         28 . (canceled) 
     
     
         29 . (canceled) 
     
     
         30 . (canceled) 
     
     
         31 . A method for treatment and/or prevention of a pathological condition involving ECM and/or cartilage degradation in a subject comprising administering to said subject a therapeutically effective amount of agent selected from the group consisting of an agent having binding affinity with a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 39, 40, 27, 42, 22-26, 28-38, and 41 and fragments thereof, and an antisense polynucleotide, a ribozyme, and a small interfering RNA (siRNA) comprising a nucleic acid sequence complementary to, or engineered from, a naturally-occurring polynucleotide sequence of about 17 to about 30 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 18, 19, 6, 21, 1-5, 7-17, and 20. 
     
     
         32 . A method for diagnosing a pathological condition involving ECM and/or cartilage degradation, comprising determining a first amount or activity of polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:39, 40, 27, 42, 22-26, 28-38, and 41 present in a biological sample obtained from said subject, and comparing said first amount or activity with the ranges of amounts or activities of the polypeptide determined in a population of healthy subjects, wherein an increase of the amount or activity of polypeptide in said biological sample compared to the range of amounts or activities determined for healthy subjects is indicative of the presence of the pathological condition. 
     
     
         33 . The method of  claim 31  wherein the pathological condition is osteoarthritis. 
     
     
         34 . The method of  claim 32  wherein the pathological condition is osteoarthritis.