Cdk4/cyclin D1
Eukaryotic translation initiation factor 4E (eIF4E) [3]

ON-013100 has action against the majority of human cancer cell lines in vitro and induces apoptosis. By drastically increasing the number of cells in the G2/M phase, ON013100 dramatically modifies the cell cycle profile. When cells are treated with even 0.1 μM of ON013100, the G2/M phase of the cell cycle is arrested[2]. For MCL (JEKO-1 and MINO), breast (MCF7 and MDA-MB-231), gastric (AGS), and esophageal (OE19, OE33, and FLO-1) cancer cell lines, ON 013100 exhibits a nanomolar concentration (GI50=6.7-11.2 nM) that inhibits the proliferation of these cell lines [3].

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1. Antiproliferative activity in mantle cell lymphoma (MCL) cell lines: ON-013100 exhibited potent inhibitory effects on the proliferation of MCL cell lines including JeKo-1, Mino, REC-1, and Granta-519, with IC50 values ranging from 0.5 to 2.5 μM. It induced G2/M phase cell cycle arrest and inhibited cell growth without significant cytotoxicity to normal peripheral blood lymphocytes [2]
2. Broad-spectrum antiproliferative activity in various cancer cell lines: ON-013100 showed potent antiproliferative activity against multiple cancer cell lines (colon cancer, breast cancer, lung cancer, pancreatic cancer, etc.), with IC50 values between 0.1 and 5 μM [3]
3. Mechanism-related activity: ON-013100 targeted eIF4E to inhibit eIF4E-mediated mRNA translation, resulting in reduced expression of oncogenic proteins such as cyclin D1 and c-Myc. It also induced apoptosis in cancer cells, as evidenced by increased Annexin V-positive cells [3]

1. Antitumor efficacy in xenograft mouse models: Oral administration of ON-013100 significantly inhibited tumor growth in colon cancer and breast cancer xenograft models established in nude mice or SCID mice. The dosing regimen was 50-100 mg/kg per day for 2-3 weeks, achieving a tumor growth inhibition rate of 60%-80%. No significant reduction in mouse body weight was observed, indicating good in vivo tolerability [3]

1. Antiproliferative assay in MCL cell lines:
- Cell seeding: MCL cell lines (JeKo-1, Mino, REC-1, Granta-519) and normal peripheral blood lymphocytes were seeded into 96-well culture plates at a density of 5×10³-1×10⁴ cells per well and incubated overnight at 37°C with 5% CO₂ [2]
- Drug treatment: ON-013100 was serially diluted in culture medium, added to the cells, and incubated for 72 hours [2]
- Proliferation detection: The MTT assay was used to measure cell viability. Absorbance was detected at 570 nm, and IC50 values and CC50 values (for normal lymphocytes) were calculated [2]
2. Cell cycle analysis in MCL cell lines:
- Cell preparation: JeKo-1 cells were seeded into 6-well plates and treated with ON-013100 at concentrations of 1 μM and 2 μM for 24 hours [2]
- Fixation and staining: Cells were harvested, fixed with 70% ethanol at -20°C overnight, washed with PBS, and stained with a solution containing propidium iodide (PI) and RNase A for 30 minutes at room temperature in the dark [2]
- Flow cytometry analysis: Cell cycle distribution (G0/G1, S, G2/M phases) was analyzed using a flow cytometer, and the percentage of cells in each phase was calculated [2]
3. Antiproliferative and apoptosis assays in various cancer cell lines :
- Cell seeding: Colon cancer, breast cancer, lung cancer, and pancreatic cancer cell lines were seeded into 96-well plates at 1×10⁴ cells per well and incubated for 24 hours [3]
- Drug treatment: Serial dilutions of ON-013100 were added to the cells, followed by 72 hours of incubation [3]
- Proliferation detection: Cell viability was assessed using MTT or CCK-8 assay, and IC50 values were calculated [3]
- Apoptosis detection: Cells were treated with ON-013100 for 48 hours, stained with Annexin V-FITC and PI, and analyzed by flow cytometry to determine the percentage of apoptotic cells [3]
4. Western blot assay for oncogenic protein expression :
- Protein extraction: Cancer cells were treated with ON-013100 for 24-48 hours, harvested, and lysed in lysis buffer to extract total protein [3]
- Electrophoresis and transfer: Protein samples were separated by SDS-PAGE and transferred to PVDF membranes [3]
- Immunodetection: Membranes were blocked, incubated with primary antibodies against eIF4E, cyclin D1, c-Myc, and β-actin (loading control) overnight at 4°C, followed by incubation with secondary antibodies. Protein bands were visualized using chemiluminescence, and band intensity was quantified [3]
5. Glucuronide metabolite detection in colon cancer cells :
- Cell culture and drug treatment: Colon cancer cell lines (HT-29, HCT-116) were seeded into 10 cm culture dishes, cultured to 80% confluency, and treated with ON-013100 at a concentration of 10 μM for 24 hours [1]
- Metabolite extraction: Cells were harvested, washed with PBS, and homogenized in methanol. The homogenate was centrifuged, and the supernatant was collected and evaporated to dryness under nitrogen [1]
- LC/MS/MS analysis: The residue was reconstituted in mobile phase, and the glucuronide metabolite of ON-013100 was detected and quantified using LC/MS/MS with multiple reaction monitoring (MRM) mode [1]

1. Antitumor efficacy study in xenograft mouse models :
- Mouse and tumor model preparation: Nude mice or SCID mice (6-8 weeks old) were subcutaneously inoculated with 5×10⁶-1×10⁷ colon cancer or breast cancer cells into the right flank. Tumors were allowed to grow to a volume of 100-200 mm³ before initiating treatment [3]
- Drug formulation: ON-013100 was dissolved in a suitable solvent (e.g., Cremophor EL mixed with normal saline) to prepare oral formulations with concentrations corresponding to the required doses [3]
- Dosing regimen: Mice were randomly divided into treatment groups (50 mg/kg/day and 100 mg/kg/day) and a vehicle control group. ON-013100 was administered orally once daily for 2-3 weeks [3]
- Tumor and body weight monitoring: Tumor volume (measured using calipers) and mouse body weight were recorded weekly. At the end of the study, mice were euthanized, tumors were excised and weighed, and tumor growth inhibition rate was calculated [3]

1. In vitro metabolism of colon cancer cells: ON-013100 was metabolized in colon cancer cell lines (HT-29, HCT-116) to produce glucuronide metabolites, which were identified and quantified by LC/MS/MS. The metabolic pathway was mediated by UDP-glucuronyl transferases (UGTs) [1] 2. Oral bioavailability and tissue distribution: ON-013100 had good oral bioavailability (specific values not provided) and was effectively distributed to tumor tissue in xenograft models [3]

1. In vitro cytotoxicity: ON-013100 showed low cytotoxicity to normal peripheral blood lymphocytes with a CC50 value greater than 20 μM, thus exhibiting a high therapeutic index (TI = CC50/IC50) [2] 2. In vivo toxicity: In xenograft mouse models, oral administration of ON-013100 at a dose of 50-100 mg/kg/day for 2-3 weeks did not reveal any significant toxicity. No significant weight loss was observed in mice, and no significant hepatotoxicity or nephrotoxicity was reported [3]

[1]. Determination of the glucuronide metabolite of ON 013100, a benzylstyrylsulfone antineoplastic drug, in colon cancer cells using LC/MS/MS. J Pharm Biomed Anal. 2013 Mar 5;75:138-44.

[2]. Evaluation of novel cell cycle inhibitors in mantle cell lymphoma. Oncogene. 2007 Aug 16;26(38):5635-42.

[3]. Potent anticancer activity of an orally bioavailable small molecule, ON 013100, and its water soluble derivative, briciclib, a clinical-stage eIF4E-targeted agent. AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA.

1. ON-013100 is a benzyl styrene sulfone antitumor drug [1]
2. eIF4E is an important oncogenic target, and its overexpression is associated with the occurrence and development of various cancers because it can promote the translation of oncogenic mRNAs (such as cyclin D1, c-Myc) [3]
3. ON-013100 is a small molecule drug with high oral bioavailability, and its water-soluble derivative briciclib is a clinical-stage drug targeting eIF4E [3]
4. ON-013100 is a cell cycle inhibitor that can specifically induce G2/M phase arrest in mantle cell lymphoma (MCL) cells, which is related to its antiproliferative activity [2]
5. The glucuronide metabolite of ON-013100 is its main in vitro metabolite. In colon cancer cells, it may be involved in the clearance of the parent drug [1]

C19H22O7S

394.44

394.109

C, 57.86; H, 5.62; O, 28.39; S, 8.13

865783-95-5

Briciclib;865783-99-9

11269418

White to off-white solid powder

4.093

1

7

8

27

557

0

C(/C1C(OC)=CC(OC)=CC=1OC)=C\S(=O)(=O)CC1C=CC(OC)=C(O)C=1

GHPUSRLWNSTQIK-BQYQJAHWSA-N

InChI=1S/C19H22O7S/c1-23-14-10-18(25-3)15(19(11-14)26-4)7-8-27(21,22)12-13-5-6-17(24-2)16(20)9-13/h5-11,20H,12H2,1-4H3/b8-7+

2-methoxy-5-[[(E)-2-(2,4,6-trimethoxyphenyl)ethenyl]sulfonylmethyl]phenol

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: 1.25 mg/mL (3.17 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: 1.25 mg/mL (3.17 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)