Cell lines

human Ovarian Cancer cell lines (HeyA8 and HeyA8MDR), Cervical cancer cell line (OV2008, C13)

Preparation Method

Cells were treated with PFK158, carboplatin (CBPt), Paclitaxel (PTX) alone and in combination, and Phosphatidyl‐serine externalization was analyzed by double staining the cells with FITC‐Annexin V and PI.

Reaction Conditions

10 μM PFK-158 for 24h.

Applications

PFK-158 treatment sensitizes chemoresistant cells (C13) and induces cell death. The combined treatment of PFK158 and CBPt resulted in significant increase in apoptosis in C13 (45%) compared to OV2008 cells (24.6%). Similar analysis using a combination of PFK158 and PTX showed a marked increase in apoptosis in the HeyA8MDR (70%) cells compared to HeyA8 (48%), respectively.

Animal models

Endometrial cancer (EC) mouse xenograft models

Preparation Method

HEC-1B and ARK-2 cells were subcutaneously injected. Following the detection of palpable tumors, the mice were treated with vehicle, PFK158 alone 2×/week, carboplatin (CBPt) alone 1×/week, or both for 14 days.

Dosage form

35mg/kg PFK-158, intraperitoneal(i.p.) injection

Applications

A significant reduction of tumor growth, tumor volume and tumor weight was observed at day 28 in both PFK158 alone and combination groups. H&E staining results demonstrated that PFK158- and combination treatment significantly increased tumor necrosis compared to control

PFK-158 is a potent and selective PFKFB3 inhibitor, which shows extensive anti-tumor activity by reducing the uptake of glucose in cancer cells, the production of ATP, the release of lactic acid and inducing apoptosis and autophagy^[1].

PFK-158 suppressed cell viability in a dose- and time-dependent manner in EC cells. Co-treatment with PFK158 (5 μM) and CBPt led to a significant increase in the percentage of apoptotic cells in HEC-1B and ARK-2. Furthermore, Western blot analysis revealed that the active form of PARP was significantly increased upon co-treatment, compared to single treatment alone, further demonstrating that the combination treatment enhances cell apoptosis^[2]

The efficacy of PFK158 alone and in combination with carboplatin (CBPt) was evaluated on primary tumor growth and metastasis in HeyA8MDR‐earing nude mice i.p. A marked reduction of tumor growth was observed in the combination treatment.PFK-158 with CBPt significantly reduced ascites and reduced LDs in tumor tissue as seen by immunofluorescence and transmission electron microscopy compared to untreated mice^[3]

References:
[1]. Gustafsson NMS, Farnegardh K, et al. Targeting PFKFB3 radiosensitizes cancer cells and suppresses homologous recombination. Nat Commun. 2018 Sep 24;9(1):3872.
[2]. Xiao Y, Jin L, et al. Inhibition of PFKFB3 induces cell death and synergistically enhances chemosensitivity in endometrial cancer. Oncogene. 2021 Feb;40(8):1409-1424.
[3]. Mondal S, Roy D, et al. Therapeutic targeting of PFKFB3 with a novel glycolytic inhibitor PFK158 promotes lipophagy and chemosensitivity in gynecologic cancers. Int J Cancer. 2019 Jan 1;144(1):178-189.