In Vivo Activation of the p53 Pathway by Small-Molecule Antagonists of MDM2

American Association for the Advancement of Science (AAAS) - Tập 303 Số 5659 - Trang 844-848 - 2004
Lyubomir T. Vassilev1,2,3, Binh Thanh Vu1,2,3, Bradford Graves1,2,3, Daisy Carvajal1,2,3, Frank Podlaski1,2,3, Zoran Filipovic1,2,3, Norman Kong1,2,3, U. Kammlott1,2,3, Christine Lukacs1,2,3, Christian Klein1,2,3, Nader Fotouhi1,2,3, Emily A. Liu1,2,3
1Department of Chemistry, Roche Research Center, Hoffmann–La Roche, Inc., Nutley, NJ 07110, USA.
2Department of Discovery Oncology, Roche Research Center, Hoffmann–La Roche, Inc., Nutley, NJ 07110, USA.
3Pharma Research, Roche Diagnostics GmbH, 82372 Penzberg, Germany.

Tóm tắt

MDM2 binds the p53 tumor suppressor protein with high affinity and negatively modulates its transcriptional activity and stability. Overexpression of MDM2, found in many human tumors, effectively impairs p53 function. Inhibition of MDM2-p53 interaction can stabilize p53 and may offer a novel strategy for cancer therapy. Here, we identify potent and selective small-molecule antagonists of MDM2 and confirm their mode of action through the crystal structures of complexes. These compounds bind MDM2 in the p53-binding pocket and activate the p53 pathway in cancer cells, leading to cell cycle arrest, apoptosis, and growth inhibition of human tumor xenografts in nude mice.

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Tài liệu tham khảo

10.1038/35042675

10.1016/S0092-8674(00)81871-1

10.1126/science.1905840

10.1038/358080a0

10.1101/gad.7.7a.1126

10.1038/362857a0

10.1002/bies.950151008

10.1038/sj.onc.1203012

10.1016/S1044-579X(02)00099-8

10.1093/nar/26.15.3453

D. Freedman, A. J. Levine, Cell. Mol. Life Sci.5, 96 (1999).

10.1038/sj.bjc.6690312

10.1038/nrc991

10.1016/S0960-9822(06)00374-5

10.1073/pnas.95.1.195

10.1038/sj.onc.1202528

10.1006/jmbi.2000.3738

10.1002/1097-0215(20001201)88:5<804::AID-IJC19>3.0.CO;2-Z

R. Zhang, H. Wang, Methods Mol. Med.85, 205 (2003).

10.1126/science.274.5289.948

The N-terminal domain of human MDM2 (residues 25 to 108) was used for crystallization studies. Leu 33 → Glu 33 mutation was introduced to increase surface charge and thus improve the solubility and crystallization properties of the protein.

10.1126/science.282.5393.1497

10.1016/0092-8674(93)90500-P

Cells with mutant p53 accumulate high levels of transcriptionally inactive protein as a result of their inability to up-regulate MDM2 transcription.

L. T. Vassilev unpublished data.

Materials and methods are available as supporting material on Science Online.

To quantitate the selectivity of MDM2 inhibitors we used a parameter defined as a ratio between the average IC 50 for two mutant p53 lines (SW480 and MDA-MB-435) and that for three wild-type p53 lines (HCT116 RKO and SJSA-1). The compounds reported here showed selectivity in the 11 to 18 range.

10.1038/sj.onc.1203013

A deletion construct of human p53 (p53C312) coveringthe N-terminal binding site and the central DNA binding domain (residues 1 to 312) but lacking the C-terminal tetramerization domain was used for Biacore binding studies.

L. T. Vassilevet al., Anticancer Drug Des.13, 359 (2001).

We thank C. Tovar G. Kaplan K. Dillon A. Specian A. Schutt C.-M. Liu B. Felix Q. Xiang R. Margolis W. Qing K. Frank H. Butscher and F. Hesse for providing reagents and experimental help; and D. Emerson D. Fry S.-S. So J. Tilley J. Roberts D. Presky H.-J. Mueller G. Ju D. Walker L. Babiss and D. Heimbrook for discussions. Coordinates are available from the Protein Data Bank with accession code 1RV1.

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American Association for the Advancement of Science (AAAS)

Tập 303 Số 5659

844-848

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