Fig 1.
Structure, biochemical activity and melt curve of JBI-097.
(1A) Chemical Structure of JBI-097. (1B) JBI-097 potently inhibited LSD1 in TR-FRET Assay. The results shown are representative from two independent experiments. (1C and 1D) Derivative melt curves of 1C. LSD1 and 1D. HDAC8 in the presence of JBI-097 or DMSO. Experiments were done in triplicates.
Table 1.
HDAC Isoform Selectivity of JBI-097.
Table 2.
Anti-proliferative activity of JBI-097 in various cancer cell lines.
Fig 2.
Anti-proliferative activity of JBI-097 in human erythroleukemia HEL 92.1.7 cells.
Cell proliferation was evaluated using a Alamar Blue assay. Human cell lines (HEL 92.1.7; density, 5 and 10 × 103 for 3 days and 6 days; respectively) were incubated with or without the indicated concentrations of JBI-097, Ricolinostat and Iadademstat for 72 and 144 h. The results shown are representative from two independent experiments.
Fig 3.
JBI-097 effect on the modulation of differentiation markers in HEL 92.1.7 cells.
(3A) Effect of JBI-097 on CD11b modulation in HEL 92.1.7 Cells. Cells were treated with 8 concentrations of JBI-097, 10 and 1 μM of JBI-135 (Ricolinostat) and JBI-236 (Iadademstat) for 24h and the mRNA level of CD11b was estimated by RT-qPCR. Error bars indicate mean ± S.E, (3B) Effect of JBI-097 on CD86 modulation in HEL 92.1.7 Cells. Cells were treated with 8 point CRC of JBI-097, 10 and 1 μM of JBI-135 (Ricolinostat) and JBI-236 (Iadademstat) for 24h and the mRNA level of CD86 was estimated by RT-q PCR. Error bars indicate mean ± S.E, (3C) Effect of JBI-097 on Gfi1b modulation in HEL 92.1.7 Cells. Cells were treated with 8 point CRC of JBI-097, 10 and 1 μM of JBI-135 (Ricolinostat) and JBI-236 (Iadademstat) for 24h and the mRNA level of Gfi1b was estimated by RT-q PCR. Error bars indicate mean ± S.E.
Fig 4.
JBI-097 inhibition increased tubulin acetylation in HEL92.1.7 cells.
HEL 92.1.7 cells were cultured with vehicle control, JBI-097 (3fold—8point dose response from 10 μM), Ricolinostat (10, 1 μM) and Iadademstat (10, 1 μM) for 3 h. Whole-cell lysates were subjected to immunoblotting with the indicated antibodies. Vinculin was used as loading control.
Fig 5.
Efficacious of JBI-097 in multiple Xenograft models.
5A: SCID mice were injected s.c. with 5 × 106 HEL 92.1.7 cells and were treated with vehicle, JBI-097 (25 and 50 mg/kg, po, QD), for 14 d.; n = 6 mice per group. 5B: HEL92.1.7 xenograft: Body weight monitored from Day1 to Day14; n = 6 mice per group. 5C: SCID mice were injected s.c. with 5 × 106 MM1.S cells and were treated with vehicle, JBI-097 (12.5 mg/kg, po), Bortezomib (0.5 mg/kg, IP, BIW), Pomalidomide (2.5mg/kg, po, 5x/week), or Bortezomib + JBI-097 or Pomalidomide + JBI-097 for 13 d.; n = 7 mice per group. 5D: MM1.S xenograft: Body weight monitored from Day1 to Day13; n = 7 mice per group. 5E: BALB/c mice were injected s.c. with 1 × 106 CT-26 cells and were treated with vehicle, JBI-097 (20 mg/kg, po), Anti PD-L1 (100μg/animal, IP, Q4D), or Anti PD-L1 + JBI-097 for 14 d.; n = 7 mice per group. 5F: CT-26 syngeneic model: Body weight data from Day1 to Day14; n = 7 mice per group. All data are represented as mean ± SE. In all the studies tumor volume was measured and calculated versus time (days).