| ln Vitro |
FKL-137 (0.001-10 μM; 24-72 hours) can effectively inhibit the proliferation of HEL and K562 erythroleukemia cells, and its safety against normal LX-2 hepatocytes is relatively high compared with HepG2 liver cancer cells [1]. FKL-137 (0.1-0.4 μM; 48 hours) can induce apoptotic nuclear changes in HEL and K562 erythroleukemia cells [1]. FKL-137 (0.1-0.4 μM; 48 hours) can induce apoptosis in HEL and K562 erythroleukemia cells in a dose-dependent manner. Significant apoptotic cell populations were detected in K562 cells at all tested concentrations, and significant apoptotic cell populations were also detected in HEL cells at concentrations of 0.2-0.4 μM [1]. FKL-137 (0.1–0.4 μM; 48 h) modulates the expression of apoptosis-related proteins in HEL and K562 erythroleukemia cells by upregulating Bax and Cleaved-PARP and downregulating Bcl-2 and PARP [1]. FKL-137 (0.1–0.4 μM; 48 h) downregulates the expression of glucose metabolism-related proteins HK2, PKM2, and LDH in HEL and K562 erythroleukemia cells [1]. FKL-137 (0.1–0.4 μM; 48 h) dose-dependently downregulates the protein levels of PI3K, p-PI3K, AKT, and p-AKT in HEL and K562 erythroleukemia cells [1]. FKL-137 (0.1-0.4 μM; 6-24 h) inhibited glucose uptake and lactate secretion in HEL and K562 erythroleukemia cells in a dose- and time-dependent manner, with significant effects observed as early as 6 h [1]. FKL-137 (0.001-10 μM; 2 h) at concentrations of 0.01-1 μM improved the thermostability of GLUT1 in K562 erythroleukemia cells without altering the total GLUT1 protein level [1]. FKL-137 (0-0.6 μM; 6 h) inhibited GLUT1-mediated glucose uptake in K562 erythroleukemia cells with an IC50 value of 0.15 μM, and its inhibitory effect was weakened in high glucose medium, confirming its competitive targeting of GLUT1 [1]. FKL-137 (0.1–0.4 μM) dose-dependently reduced GLUT1 expression in cultured HEL erythroleukemia cells[1]. Compared with the negative control group, FKL-137 (0.001–10 μM; 48–72 h) showed significantly stronger dose- and time-dependent antiproliferative effects on GLUT1 knockdown K562 erythroleukemia cells at 72 h[1]. FKL-137 (0.1–0.4 μM; 6–24 h) further inhibited glucose uptake and lactate secretion in GLUT1 knockdown K562 erythroleukemia cells, with significant effects observed within 12 h[1]. FKL-137 (0.1–0.4 μM; 48 h) regulated the compensatory upregulation of glucose metabolism-related proteins HK2 and LDH in GLUT1 knockdown K562 erythroleukemia cells[1].
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| ln Vivo |
FKL-137 (1-10 mg/kg; intraperitoneal injection; once every 48 hours; for a total of 7 times) can dose-dependently increase hematocrit, reduce splenomegaly and organ ratio, improve histopathological damage, reduce splenic glucose and lactate levels, and downregulate metabolic proteins in Fr-MuLV-induced erythroleukemia mice, with the 10 mg/kg dose producing the most significant effect (hematocrit, spleen weight and glucose content p<0.001 compared with the model group)[1].
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| Cell Assay |
Cell viability assay [1]
Cell Types: Human erythroleukemia HEL cells, human erythroleukemia K562 cells, human normal hepatic stellate cells LX-2 cells, human hepatocellular carcinoma HepG2 cells Tested Concentrations: 0.001 μM; 0.01 μM; 0.1 μM; 1 μM; 10 μM Incubation Duration: 24 hours, 48 hours, 72 hours Experimental Results: It showed significant, time- and dose-dependent inhibitory effects on the proliferation of HEL and K562 cells. In HEL cells, the IC50 values were 0.6 μmol/L (24 hours), 0.25 μmol/L (48 hours), and 0.1 μmol/L (72 hours), respectively. In K562 cells, the IC50 values were 0.8 μmol/L (24 h), 0.4 μmol/L (48 h), and 0.2 μmol/L (72 h), respectively. The hepatotoxicity safety index values were 1.65 (24 h), 1.66 (48 h), and 2.29 (72 h), respectively, indicating that it has relative safety. Apoptosis Analysis [1] Cell Types: Human erythroleukemia HEL cells, Human erythroleukemia K562 cells Tested Concentrations: 0.1 μM; 0.2 μM; 0.4 μM Incubation Duration: 48 hours Experimental Results: Characteristic apoptotic morphological changes, including nuclear condensation, nuclear fragmentation, and chromatin marginalization, were induced in both HEL and K562 cells. The effect was enhanced with increasing compound concentration. The number of apoptotic cells increased in a dose-dependent manner in both cell lines. In HEL cells, concentrations of 0.2 μM and 0.4 μM significantly increased the early and late apoptosis rates. At all tested concentrations, the early and late apoptosis rates were significantly increased, with the most significant effect observed at 0.4 μM in K562 cells. Western Blot Analysis [1] Cell Types: Human erythroleukemia HEL cells, Human erythroleukemia K562 cells Tested Concentrations: 0.1 μM; 0.2 μM; 0.4 μM Incubation Duration: 48 hours Experimental Results: Compared with the DMSO control group, the expression of pro-apoptotic proteins Bax and Cleaved-PARP in HEL and K562 cells was upregulated in a dose-dependent manner. Compared with the DMSO control group, the expression of anti-apoptotic proteins Bcl-2 and full-length PARP protein in HEL and K562 cells was downregulated. Compared with the DMSO control group, the expression of glucose metabolism-related proteins HK2, PKM2 and LDH in HEL and K562 cells was downregulated in a dose-dependent manner. The protein levels of PI3K, p-PI3K, AKT, and p-AKT in HEL and K562 cells were downregulated in a dose-dependent manner.
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| Animal Protocol |
Animal/Disease Models:BALB/c (8-10 week old males and females; Fr-MuLV-induced erythroleukemia model established at 6 weeks of age)[1]
Doses: 1 mg/kg; 5 mg/kg; 10 mg/kg Route of Administration: Intraperitoneal injection; every 48 hours; 7 doses in total Experimental Results: Hematocrit increased to approximately 42% at 1 mg/kg, to approximately 43% at 5 mg/kg, and to approximately 55% at 10 mg/kg. Spleen weight decreased to approximately 0.9 g at 1 mg/kg, to approximately 0.4 g at 5 mg/kg, and to approximately 0.2 g at 10 mg/kg. All doses significantly reduced the organ proportions of the liver and spleen; no significant changes were observed in the organ proportions of the heart, lungs, or kidneys. Compared with the model group, pathological mitotic figures and tumor cell infiltration foci were significantly reduced in spleen tissue in all dose groups. Compared with the model group, leukocyte infiltration was significantly reduced in liver tissue in all dose groups; no significant histopathological changes were observed in heart, lung, or kidney tissues. In the 1 mg/kg dose group, spleen glucose levels decreased to approximately 0.22 mmol/g and lactate levels decreased to approximately 0.28 mmol/g; in the 5 mg/kg dose group, spleen glucose levels decreased to approximately 0.12 mmol/g and lactate levels decreased to approximately 0.3 mmol/g; and in the 10 mg/kg dose group, spleen glucose levels decreased to approximately 0.08 mmol/g and lactate levels decreased to approximately 0.3 mmol/g. Compared with the model group, the expression of HK2 and LDH proteins was downregulated in spleen tissue in all dose groups. |
| References |
| Appearance |
Typically exists as solids at room temperature
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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.) |
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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