2022 Fiscal Year Annual Research Report
STAT3を分解するPROTAC/SNIPERの分子設計、合成と活性評価
| Project/Area Number |
22F22107
|
|---|
| Allocation Type | Single-year Grants |
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
内藤 幹彦 東京大学, 大学院薬学系研究科(薬学部), 特任教授 (00198011)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SHIH PO-CHANG 東京大学, 薬学研究科(研究院), 外国人特別研究員
|
|---|
| Project Period (FY) |
2022-04-22 – 2024-03-31
|
| Keywords | PROTAC / STAT3 / SNIPER / protein degradation |
|---|
| Outline of Annual Research Achievements |
Proteolysis targeting chimera (PROTAC) is a technology that uses chimeric chemicals to bind to a protein of interest (POI) and an E3 ligase so as to induce POI degradation through the ubiquitin-proteasome system. In this project, we intended to develop novel PROTAC degraders targeting STAT3.
First, we employed sulfonamide derivatives that are believed to bind to the SH2 domain of STAT3 as small molecule-warheads and synthesized PROTACs by conjugating to thalidomide (a cereblon E3 ligase binder) via alkyl linkers. The molecular weights of the resulting PROTACs are approximately 900. Then, the STAT3 degradation activity of these PROTACs were tested along with their parental STAT3 binders in MCF7 and SU-DHL-1 cells. The western blot analysis indicated that PS314 (PROTAC, using PS313 as a warhead) showed a significant activity to reduce STAT3 protein level in both cell lines, while PS312 (PROTAC, using PS311 as a warhead) did not reduce the STAT3 protein level.
We also developed oligonucleotide-warheaded PROTAC molecules against STAT3. A decoy oligonucleotide to which STAT3 binds was conjugated to three different E3 ligands to be LCL-STAT3 decoy, POM-STAT3 decoy and VH-STAT3 decoy. Tested in MCF7 cells, POM-STAT3 decoy was found to be more able to degrade STAT3 than VH-STAT3 decoy, and LCL-STAT3 decoy showed no degradation activity.
|
| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
We designed various PROTACs/SNIPERs that are categorized into two groups, one with small molecule binders and the other with decoy oligonucleotide to which STAT3 binds. The chimeric molecules were smoothly synthesized and some of the PROTACs showed an activity to reduce STAT3 protein level in cancer cells.
|
| Strategy for Future Research Activity |
(1) The degradation activity of PROTACs against STAT3 protein will be convincingly demonstrated by repeated experiments.
(2) The degradation activity of PROTACs will be tested in combination with various inhibitors (E1, E3, proteasome and STAT3), to validate degradation mechanisms.
(3) The active PROTACs will be evaluated for their activity to inhibit growth of cancer cells that are dependent on STAT3, such as lung cancer cells NCI-H2087 and lymphoid cells MOLM16.
|
