Methods and means for treatment of osteoarthritis
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 type II collagen degradation in cartilage or extracellular matrix (ECM), said method comprising:
a) contacting a compound with a polypeptide comprising the amino acid sequence SEQ ID NO: 27;
b) determining the binding affinity of the compound to the polypeptide;
c) contacting a population of mammalian cells expressing the polypeptide with compound that exhibits a binding affinity in (b) of at least 10 micromolar; and
d) identifying a compound that reduces the expression of a cartilage degradation protein or protease in the mammalian cells or inhibits the induction of expression of a cartilage degradation protein or protease in the mammalian cells in response to IL-1,
wherein said cartilage degradation protein or protease is MMP13.
2. A method for identifying a compound that inhibits type II collagen degradation in cartilage or extracellular matrix (ECM), said method comprising:
a) contacting a compound with a polypeptide comprising the amino acid sequence SEQ ID NO: 27;
b) determining the ability of the compound to inhibit the expression or activity of the polypeptide;
c) contacting a population of mammalian cells expressing the polypeptide with compound that significantly inhibits the expression or activity of the polypeptide; and
d) identifying a compound that reduces expression of a cartilage degradation protein or protease in the mammalian cells or inhibits the induction of expression of a cartilage degradation protein or protease in the mammalian cells in response to IL-1,
wherein said cartilage degradation protein or protease is MMP13.
3. The method according to any one of claim 1 or 2 , which additionally comprises the step of comparing the compound to be tested to a control.
4. The method according to claim 3 , wherein the control is where the polypeptide has not been contacted with the compound.
5. The method according to claim 1 or 2 , which additionally comprises the step of comparing the compound to a control, wherein the control is a population of mammalian cells that does not express said polypeptide.
6. The method according to any one of claim 1 , 2 , or 3 - 5 , wherein the compound is selected from the group consisting of compounds of a commercially available screening library and compounds having binding affinity for a polypeptide comprising the amino acid sequence SEQ ID NO: 27.
7. The method according to claims 1 - 2 , wherein the compound is a peptide in a phage display library or an antibody fragment library.