Methods of identifying modulators of kinases responsive to stress

• US 6,416,964 B2
• Filed: 01/27/1999
• Issued: 07/09/2002
• Est. Priority Date: 01/29/1998
• Status: Expired due to Fees

• Current Assignee: Merck & Co., Inc.
• Original Assignee: Merck & Co., Inc.
• Inventors: Reszka, Alfred A., Rodan, Gideon A.
• Primary Examiner(s): Lankford, Jr., Leon B.
• Application Number:
  US09/238,536
• Publication Number:
  US 20010014329A1
• Time in Patent Office: 1,259 Days
• Field of Search: 435/4, 435/194, 435/15, 435/325, 435/366
• US Class Current: 435/15
• CPC Class Codes: A61K 31/663 Compounds having two or more phosphorus acid groups or esters thereof, e.g. clodronic acid, pamidronic acid
• CPC Class Codes: C12Q 1/485 involving kinase
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5044 involving specific cell types

Abstract

The present invention relates to methods of identifying compounds useful as modulators of certain stress responsive kinases. More particularly, the compounds so identified are useful for treating or preventing diseases or conditions that are mediated by, for example, abnormal bone resorption or angiogenesis. These compounds are useful for treating or preventing osteoporosis, and inhibiting vascular restenosis, diabetic retinopathy, macular degeneration, angiogenesis, atherosclerosis, inflammation, and tumor growth.

Claims

1. A method for identifying compounds which are useful for treating abnormal bone resorption or angiogenesis comprising:

(a). contacting a putative activity-modifying compound with a cell culture;

(b). determining the kinase activity of said cell culture and comparing said kinase activity with a cell culture not contacted with said putative activity-modifying compound; and

(c). identifying a compound which activates a kinase response to stress thereby identifying a compound useful for treating abnormal bone resorption or angiogenesis.

2. A method according to claim 1 wherein said kinase responsive to stress is selected from the group consisting of Mst 1, Mst 2, 34 kDa kinase which regulates osteoclast function and is an amino terminal clevage product of Mst, 50 kDa kinase which regulates osteoclast function and is activated by echistatin or an organic bisphosphonate and 130 kDa kinase which regulates osteoclast function and is activated by echistatin or an organic biphosphonate, and combinations thereof.

3. A method according to claim 2 wherein said cell culture is selected from the group consisting of an osteoclastogenesis cell culture, purified osteoclasts, partially-purified osteoclasts, unpurified osteoclasts, purified pre-osteoclasts, partially-purified pre-osteoclasts, unpurified pre-osteoclasts, purified osteoclast-like cells, partially-purified osteoclast-like cells, unpurified osteoclasts-like cells, and mixtures thereof.

4. A method according to claim 3 wherein said kinase activity is determined using an in-gel kinase assay.