| Size | Price | |
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| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
DDX5 (p68) – FL77-24 binds to and induces degradation of the DDX5 oncoprotein, which is a master regulator controlling multiple downstream targets including survivin, Mcl-1, XIAP, cIAP2, c-Myc, and mutant Kras. The compound also induces ROS production, MMP depolarization, and activation of caspases leading to apoptosis [1].
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| ln Vitro |
In Vitro: FL77-24 exhibited potent antiproliferative activity against five human cancer cell lines: HCT116 (colorectal cancer) with IC50 < 6.4 nM, HepG2 (liver cancer) with IC50 = 118.0 ± 2.8 nM, MCF-7 (breast cancer) with IC50 < 6.4 nM, A549 (lung cancer) with IC50 = 28.5 ± 6.2 nM, and HeLa (cervical cancer) with IC50 < 6.4 nM [1].
In HCT116 cells, FL77-24 induced apoptosis in a concentration-dependent manner: at 2.5 nM, early apoptosis 20.76% and late apoptosis 5.58%; at 5 nM, early apoptosis 26.86% and late apoptosis 9.79%; at 10 nM, early apoptosis 30.20% and late apoptosis 10.87%. Cell cycle analysis showed that FL77-24 treatment mainly caused cell cycle arrest in S and G2/M phases [1]. FL77-24 induced ROS production in HCT116 cells in a time- and concentration-dependent manner. ROS levels temporarily increased within 4-8 h after treatment, decreased after 12 h, and then gradually increased again. ROS production was dependent on drug concentration [1]. FL77-24 reduced mitochondrial membrane potential (MMP) in HCT116 cells: at 2.5 nM, MMP decreased to 88.86%; at 5 nM, to 82.28%; at 10 nM, to 76.37%; at 20 nM, to 71.64% (compared to 97.6% in untreated cells). The red/green fluorescence ratio decreased from 44.98 (0 nM) to 2.62 (20 nM) [1]. Western blot analysis showed that FL77-24 treatment upregulated Bax, Cyt-C, and p21 expression while downregulating Bcl-2 expression in a dose-dependent manner. The compound also downregulated survivin, XIAP, and Mcl-1 protein levels. Caspase activity assays demonstrated that FL77-24 activated caspase-3, caspase-8, and caspase-9 in a concentration-dependent manner [1]. In TOP1 enzyme inhibition assays, FL77-24 inhibited TOP1 activity only at very high concentrations (100, 50, and 25 μM) that are not physiologically relevant, indicating that TOP1 is not the primary therapeutic target [1]. RNA-Seq analysis revealed that FL77-24 modulates mRNA expression of DDX5 and topoisomerase family proteins (TOP1, TOP2A, TOP2B, TOP3A, TOP3B), activates innate immunity genes (CASP1, CXCR2, IFIH1, IL12A, IL-12B, IL-12RB2, IL-1A, IRF1, 3, 5, 7), and reduces glucose, nucleotide, and lipid metabolism gene expression [1]. |
| ln Vivo |
In Vivo: In human colorectal cancer PDX27454 tumor models, FL77-24 was administered orally at doses of 4, 8, 12, 16, and 20 mg/kg once weekly for 4 weeks (weekly × 4). At 20 mg/kg oral, the compound effectively regressed tumors. When administered intraperitoneally at doses of 4, 8, 12, 15, and 20 mg/kg once weekly for 4 weeks, FL77-24 appeared to be most effective via ip administration compared to oral administration. The compound showed good antitumor activity across a wide dose range with acceptable toxicity. The maximum tolerated dose (MTD) for oral administration appears to be at least 20 mg/kg, and for ip administration at least 15 mg/kg (did not appear to reach MTD) [1].
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| Enzyme Assay |
Enzyme Assay: For TOP1 enzyme inhibition assay, pBR322 DNA was presented as supercoiled (SC) without TOP1 treatment and relaxed (RLX) with TOP1 treatment. FL77-24 was tested at concentrations of 100, 50, 25, and 2.5 μM. The compound inhibited TOP1 activity only at the highest concentrations (100, 50, and 25 μM), with no inhibition observed at 2.5 μM. This indicates that FL77-24 does not use TOP1 as its primary therapeutic target [1].
For caspase activity assays, HCT116 cells were treated with FL77-24 for 48 h. Cell lysates (100 μg protein) were mixed with reaction buffer and chromogenic substrates (Ac-DEVD-pNA for caspase-3, Ac-IETD-pNA for caspase-8, Ac-LEHD-pNA for caspase-9), incubated at 37°C for 1 h, and absorbance was measured at 405 nm. Caspase-3, caspase-8, and caspase-9 were activated in a concentration-dependent manner [1]. |
| Cell Assay |
Cell Assay: For cell viability (MTT assay), cells (HCT116, HepG2, MCF-7, A549, HeLa) were seeded in 96-well plates, treated with various concentrations of FL77-24 for 72 h, followed by MTT addition and absorbance measurement at 570 nm. IC50 values were calculated [1].
For apoptosis and cell cycle analysis, HCT116 cells were treated with FL77-24 at 0, 2.5, 5, and 10 nM for 48 h. Cells were stained with Annexin V-FITC/PI for apoptosis or with PI for cell cycle analysis, and analyzed by flow cytometry [1]. For ROS measurement, HCT116 cells were treated with FL77-24, incubated with DCFH-DA (10 μM) for 20 min at 37°C, and fluorescence was measured at Ex/Em 488/525 nm [1]. For MMP measurement (JC-1 assay), HCT116 cells were treated with FL77-24 (2.5, 5, 10, 20 nM) for 48 h, incubated with JC-1 (4 μg/mL) for 30 min in the dark, and analyzed by flow cytometry [1]. For Western blot, HCT116 cells were treated with FL77-24 for 48 h, lysed in RIPA buffer, proteins separated by SDS-PAGE, transferred to PVDF membranes, probed with primary antibodies (Bax, Bcl-2, Cyt-C, p21, survivin, XIAP, Mcl-1, β-actin), and visualized by ECL [1]. For RNA-Seq, HCT116 cells were treated with FL77-24 (10, 50, 500 nM for 6 h; 50 nM for 6 and 24 h; 10 nM for 48 h). Total RNA was isolated using Qiazol reagent and miRNeasy Mini kit, and libraries were prepared using mRNA HyperPrep kit for NovaSeq6000 sequencing [1]. |
| ADME/Pharmacokinetics |
No direct ADME or pharmacokinetic data for FL77-24 (absorption, distribution, metabolism, excretion, half-life, oral bioavailability) were reported in the provided literature. However, based on the parent compound FL118, FL118 is rapidly cleared from circulation and effectively accumulates in tumors with a long elimination half-life [1].
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| Toxicity/Toxicokinetics |
In human PDX tumor models, FL77-24 administered orally at 4-20 mg/kg once weekly for 4 weeks showed acceptable toxicity with no clear clinical toxicity such as abnormal body weight loss out of the normal range. The MTD for oral administration appears to be at least 20 mg/kg (did not appear to reach MTD). For ip administration at 15 mg/kg, the compound also showed acceptable toxicity. At 20 mg/kg ip, no clear toxicity was observed, indicating the MTD may be higher than 15 mg/kg. The greatest advantage of FL77-24 is that it may have a high MTD, making it a candidate for pediatric cancer patients who require less toxic drugs [1].
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| References | |
| Additional Infomation |
FL77-24 is a 7-position substituted FL118 derivative with a 3,5-dimethoxyphenyl group. It was identified through screening of 24 FL118 Position 7-derived compounds and showed better antitumor activity than the parental platform drug FL118. The compound's mechanism of action is similar to FL118, targeting DDX5/p68 oncoprotein through binding and inducing its degradation. FL77-24 exhibits high antitumor activity across multiple cancer cell lines, with IC50 values < 6.4 nM against HCT116, MCF-7, and HeLa cells. RNA-Seq analysis confirmed that FL77-24 modulates similar pathways as FL118, including apoptosis, DNA repair, innate immunity activation, and metabolic control. FL77-24 has potential to be developed into an anticancer drug for pediatric cancer patients due to its low toxicity with good antitumor activity [1].
Human colorectal cancer PDX27454 tumors were maintained on SCID mice. For experiments, tumor fragments (30-40 mg) were subcutaneously transplanted into female SCID mice. When tumors reached 100-250 mm³ (day 0), mice were randomly divided into groups (2-5 mice per group). FL77-24 was formulated in organic solvent-free format with HPβCD as drug carrier in 2% HPMC and 1% PG (propylene glycol) in saline (a thick aqueous suspension) for oral (po) or intraperitoneal (ip) administration. Doses used: oral – 4, 8, 12, 16, 20 mg/kg; ip – 4, 8, 12, 15, 20 mg/kg. Treatment schedule was once weekly for 4 weeks (weekly × 4). Tumor volume was calculated as v = 0.5 × (L × W²). Body weight was monitored to assess toxicity [1]. |
| Molecular Formula |
C29H24N2O8
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|---|---|
| Molecular Weight |
528.51
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| CAS # |
2413582-39-3
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| Appearance |
Typically exists as solid at room temperature
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| Synonyms |
7-(4-Ethylphenyl)-FL118
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
Typically soluble in DMSO (e.g. 10 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8921 mL | 9.4606 mL | 18.9211 mL | |
| 5 mM | 0.3784 mL | 1.8921 mL | 3.7842 mL | |
| 10 mM | 0.1892 mL | 0.9461 mL | 1.8921 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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