Cell lines

Bacterial lipopolysaccharide (LPS)-primed macrophages prepared from adult male C57BL/6 mice(Harlan)LPS-primed THP-1 cells(to induce pro-IL-1β expression before nigericin treatment)

Preparation method

The solubility of this compound in DMSO is >21mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

1μM

Applications

Pretreatment with NSC 687852 inhibited ATP-induced IL-1β release from LPS-primed peritoneal macrophages and nigericin-induced release from LPS-primed THP-1 cells and reduced the levels of cell death induced by nigericin treatment in THP-1 cells. In macrophages, NSC 687852 also caused an increase in polyubiquitinated proteasomal substrates. In LPS-primed THP-1 cells, NSC 687852 significantly reduced the numbers of ASC specks formed after nigericin treatment. Similarly, ATP-induced speck formation in murine peritoneal macrophages was also inhibited by NSC 687852.

Animal models

combined immunodeficiency (SCID) mice with FaDu squamous carcinoma xenografts;mice with HCT-116 colon carcinoma xenografts overexpressing BCL2

Dosage form

daily subcutaneous injection ;5 mg per kg of body weight

Application

When administered NSC 687852 daily to severe combined immunodeficiency (SCID) mice with FaDu squamous carcinoma xenografts, there was a significant antitumor activity. When analyzed tumor death by measuring xenograft-derived CK18 in circulation, there was a significant increase in the plasma concentrations of total CK18 as well as increased concentrations of caspase-cleaved CK18 (CK18-Asp396) , showing that NSC 687852 had activity against tumor cells in vivo. When also examined disease-free survival in mice with HCT-116 colon carcinoma xenografts overexpressing BCL2, NSC 687852 treatment significantly delayed tumor onset compared to vehicle-treated controls, with two out of six of the mice treated with NSC 687852 being completely disease free at the end of the study.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

IC50: N/A

NSC 687852 is a 19S regulatory particle inhibitor.

The 19S particles bind polyubiquitin-linked polypeptides and present them to the 20S degradative units. USP14 and UCHL5 are cysteine enzymes that become activated after being associated with the proteasome.

In vitro: NSC 687852 blocked deubiquitylating activity of USP14 and UCHL5 selectively without inhibiting proteasome activity. NSC 687852 decreased viability in multiple myeloma (MM) cell lines and patient MM cells, inhibited MM cell proliferation even in the presence of bone marrow stroma cells, and overcomed bortezomib resistance. Anti-MM activity of NSC 687852 was associated with growth arrest through downregulating CDC2, CDC25C, and cyclin B1, as well as induction of caspase-dependent apoptosis and activation of unfolded protein response [1].

In vivo: In vivo studies using distinct human MM xenograft models showed that NSC 687852 was well tolerated, inhibited tumor growth, and prolonged mouse survival. Combination of NSC 687852 with suberoylanilide hydroxamic acid, lenalidomide, or dexamethasone was found to induce synergistic anti-MM activity [1].

Clinical trial: N/A

Reference:
[1] Ze Tian,Padraig D'Arcy,Xin Wang,Arghya Ray,Yu-Tzu Tai,Yiguo Hu,Ruben D Carrasco,Paul Richardson,Stig Linder,Dharminder Chauhan,Kenneth C Anderson.  A novel small molecule inhibitor of deubiquitylating enzyme USP14 and UCHL5 induces apoptosis in multiple myeloma and overcomes bortezomib resistance. Blood. 2014 Jan 30; 123(5): 706–716.