| Summary: |
Ubiquitin is a protein consisting
of 76 amino acids and is highly conserved throughout the tissues of
most eukaryotic organisms. The addition of ubiquitin to protein substrates
tags them for important biological processes such as degradation via
proteasome or lysosomes, cellular activity, immune response, DNA repair,
and so forth. In order to recycle ubiquitin molecules or edit polyubiquitin
chains, deubiquitinating enzymes (DUBs) hydrolyze ubiquitin bound
proteins and inhibit the proteasome degradation pathway. The cysteine
proteases ubiquitin-specific proteases (USP25 and USP28) are paralogues
of each other and have three distinct domains: an N-terminal, a pair
of ubiquitin-interacting motifs (UIM), and a ubiquitin-specific protease
(USP). |
| USP25 and USP28 have several important
cellular functions; the major role involves the regulation of the
stability of cellular proteins. USP25 serves as a tumor-promoting
factor and regulates cellular responses related to inflammation, autoimmune
disease, infectious diseases, and so forth. However, USP28 serves
to stabilize many oncoproteins and as a tumor-promoting factor. |
| Many cancer types, including breast cancer, acute
myeloid leukemia (AML), colorectal cancer, and ovarian cancer, involve
amplification, deletions, and mutations of USP28. In addition, USP28
controls the stability of MYC, a transcription factor of genes involved
in cell growth, proliferation, and apoptosis. There are studies that
showed the knockdown of USP28 gene has led to a decrease of MYC protein
and the inhibition of growth in human cell lines in vitro. Knockout
of USP28 in mice has shown that its activity is not required for normal
development and reproduction. |
| Consequently,
inhibition of USP25 and USP28 with small molecule inhibitors has potential
in the treatment of cancers, inflammatory diseases, infectious diseases,
autoimmune diseases, and so forth. The compounds of formula I in this
patent application are described to possess inhibitory activity for
USP25 and USP28 and may provide potential treatment for many of the
above disorders, particularly in cancers, autoimmune diseases, and
infectious diseases. |
| Important Compound
Classes: |
 |
| Key Structures: |
 |
| Biological
Assay: |
Ubiquitin-Rhodamine 110 assay for USP28
Activity: The final volume of the assay buffer was 20 μL containing
20 mM Tris-HCl, 2 mM CaCl2, 2 mM BME (2-mercaptoethanol),
0.01% Prionex, and 0.01% Triton X-100. Enzyme USP28, construct USP28
(USP28-5(1–1077)-TEV-6*His), substrate (Ub-Rh110; Ubiquitin-Rhodamine
110), and fluorescence were read on Envision (excitation at 485 nm
and emission at 535 nm) or on the PheraSTAR. |
| Ubiquitin-Rhodamine 110 assay for USP25 Activity: The final volume
of the assay buffer was 9 μL containing 20 mM Tris-HCl, 3 mM
BME (2-mercaptoethanol), 0.03% BGG, and 0.01% Triton X-100. Enzyme
USP25, construct USP25-His6, substrate (Ub-Rh110; Ubiquitin-Rhodamine
110), and fluorescence were read on the Envision or on the PheraSTAR. |
| Biological Data: |
In the
Table below; ++++ indicates IC50 < 0.2 μM, +++
indicates IC50 between 0.2 and 2 μM, and ++ indicates
IC50 between 2 and 10 μM.
|
| Claims: |
48 Total claims |
| 26 Composition of matter claims |
| 22 Method of use claims |
| Recent Review Articles: |
1. Kitazawa S.; Yagi-Utsumi M.; Kato K.; Kitahara R.. Molecules 2017, 22, 1414. |
| 2. Ciana A.; Achilli C.; Minetti G.. Cell. Physiol. Biochem. 2017, 42, 1139. |
| 3. Morreale F. E.; Testa A.; Chaugule V. K.; Bortoluzzi A.; Ciulli A.; Walden H.. J. Med. Chem. 2017, 60, 8183. |
| 4. Chen H.; Wu G.; Gao S.; Guo R.; Zhao Z.; Yuan H.; Liu S.; Wu J.; Lu X.; Yuan X.; Yu Z.; Zu X.; Xie N.; Yang N.; Hu Z.; Sun Q.; Zhang W.. J. Med. Chem. 2017, 60, 6828. |
| 5. Hrdinka M.; Gyrd-Hansen M.. Molecular Cell 2017, 68, 265. |
