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).</p

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.</p

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.</p

Anti-proliferative activity of JBI-097 in various cancer cell lines.

Anti-proliferative activity of JBI-097 in various cancer cell lines.</p

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.</p

S1 Checklist -

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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.</p

Raw images: Original full western blots corresponding to Fig 4.

HEL 92.1.7 cells were cultured and treated with vehicle control, JBI-097 (3fold—8point dose response from 10 μM), JBI-135 (10, 1 μM) and JBI-236 (10, 1 μM) for 3 h. Whole-cell lysates were subjected to immunoblotting with the acetyl-α-tubulin (top panels). Vinculin was used as loading control (bottom panels). Arrows indicate the bands representing Acetyl-α-tubulin and Vinculin. (PDF)</p

HDAC Isoform Selectivity of JBI-097.

HDAC Isoform Selectivity of JBI-097.</p

A New Series of Orally Bioavailable Chemokine Receptor 9 (CCR9) Antagonists; Possible Agents for the Treatment of Inflammatory Bowel Disease

Chemokine receptor 9 (CCR9), a cell surface chemokine receptor which belongs to the G protein-coupled receptor, 7-trans-membrane superfamily, is expressed on lymphocytes in the circulation and is the key chemokine receptor that enables these cells to target the intestine. It has been proposed that CCR9 antagonism represents a means to prevent the aberrant immune response of inflammatory bowel disease in a localized and disease specific manner and one which is accessible to small molecule approaches. One possible reason why clinical studies with vercirnon, a prototype CCR9 antagonist, were not successful may be due to a relatively poor pharmacokinetic (PK) profile for the molecule. We wish to describe work aimed at producing new, orally active CCR9 antagonists based on the 1,3-dioxoisoindoline skeleton. This study led to a number of compounds that were potent in the nanomolar range and which, on optimization, resulted in several possible preclinical development candidates with excellent PK properties