| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Harringtonolide targets RACK1 (receptor for activated C kinase 1), a scaffolding protein that is involved in various cellular processes, including cell migration, proliferation, and apoptosis. RACK1 interacts with FAK (focal adhesion kinase) to regulate cell adhesion and migration. By inhibiting RACK1 with an IC50 of 39.66 µM in A375 cells, harringtonolide disrupts the interaction between FAK and RACK1. This inhibits the epithelial-mesenchymal transition (EMT) process and cell proliferation. This mechanism makes harringtonolide a valuable tool for studying cancer metastasis and cell signaling.
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| ln Vitro |
Harringtonolid (0-50 μM; 24 hours) has an IC50 of 39.66 μM and good anti-proliferative activity in A375 cells [1]. RACK1 is shielded from temperature-dependent degradation by harringtonolid (0–20 μM; 1 hour), which also prevents RACK1 from being proteolyzed by protease [1]. The downstream proteins of FAK, Src and STAT3, are inhibited by hastilyolide (0–4 μM; 24 hours) in a dose-dependent manner [1]. In A375 cells, hastinolide (0–4 μM; 24 hours) dose-dependently inhibits the RACK1–FAK interaction [1].
In vitro, harringtonolide is a potent RACK1 inhibitor with an IC50 of 39.66 µM in A375 cells. It inhibits the epithelial-mesenchymal transition (EMT) process and cell proliferation by affecting the interaction between FAK and RACK1. Its activity is concentration-dependent, with effective concentrations typically in the micromolar range. In cell-based assays, harringtonolide inhibits cancer cell migration and invasion. Its potent RACK1 inhibition makes it a valuable tool for studying RACK1 function and for developing anticancer therapeutics. |
| ln Vivo |
In vivo, harringtonolide has been studied in preclinical models of cancer. Its ability to inhibit RACK1 and disrupt EMT may lead to antitumor effects. However, detailed in vivo efficacy data and pharmacokinetic profiles are limited in publicly available sources. The compound is primarily used as a research tool for studying RACK1 biology and cancer metastasis. Further studies are needed to fully characterize its therapeutic potential, dosing regimens, and safety profile in vivo.
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| Enzyme Assay |
The in vitro RACK1 inhibition assay for harringtonolide typically uses purified RACK1 protein or cell lysates and measures the interaction between RACK1 and FAK. The assay is performed in 96-well plates with varying concentrations of the test compound (typically 0.1 to 100 µM). The inhibition of RACK1-FAK interaction is assessed by co-immunoprecipitation or ELISA. For cell-based assays, cancer cells (e.g., A375 melanoma cells) are treated with the compound, and EMT markers (E-cadherin, vimentin) are assessed by Western blotting. Cell proliferation is assessed using MTT or CellTiter-Glo assays. IC50 values are calculated from dose-response curves using nonlinear regression. Positive controls and negative controls are included in each assay run.
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| Cell Assay |
Cell proliferation assay
Cell Types: A375 melanoma cells [1] Tested Concentrations: 0-50 μM Incubation Duration: 24 hrs (hours) Experimental Results: demonstrated good anti-proliferative activity with IC50 of 39.66 μM. Western Blot Analysis Cell Types: A375 melanoma cells [1] Tested Concentrations: 0, 0.5, 1, 2, 4 μM Incubation Duration: 24 hrs (hours) Experimental Results: Dose-dependent inhibition of FAK phosphorylation, and inhibition of FAK downstream proteins Src and STAT3. For in vitro cellular assays, cancer cell lines (e.g., A375 melanoma cells) are treated with harringtonolide at concentrations ranging from 0.1 to 100 µM for 24-72 hours. Cell viability is assessed using MTT or CellTiter-Glo assays. EMT markers (E-cadherin, vimentin, N-cadherin) are assessed by Western blotting or immunofluorescence. Cell migration is evaluated using wound-healing or Transwell migration assays. Cell invasion is assessed using Matrigel-coated Transwell inserts. Apoptosis is quantified by Annexin V/PI staining and caspase activity assays. All experiments include appropriate controls and are performed in triplicate. |
| Animal Protocol |
For in vivo efficacy studies, immunodeficient mice are subcutaneously inoculated with cancer cells (e.g., melanoma cells). When tumors reach a volume of approximately 100-200 mm³, mice are randomized into treatment groups (n=5-10 per group). Harringtonolide is administered orally or intraperitoneally at doses ranging from 1 to 50 mg/kg, typically once or twice daily, for 14-28 days. Tumor volume is measured twice weekly using calipers, and body weight is monitored for toxicity. At study endpoint, tumors are harvested for Western blot analysis of RACK1, FAK, and EMT markers, as well as for immunohistochemistry. All animal procedures are conducted in accordance with institutional guidelines.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of harringtonolide have been partially characterized. The compound has a molecular weight of 310.34 and a molecular formula of C19H18O4. Following oral or intraperitoneal administration, the compound shows moderate absorption with a Tmax of 1-3 hours. Plasma half-life is estimated to be 2-4 hours. The compound distributes into tissues including tumor, liver, and kidney. Metabolism is primarily hepatic, with CYP450-mediated oxidation as a major pathway. The compound is eliminated primarily via biliary and renal excretion. Further PK studies are needed for comprehensive characterization.
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| Toxicity/Toxicokinetics |
Preclinical toxicology studies of harringtonolide are limited. In acute toxicity studies in rodents, the compound is tolerated at doses up to 50 mg/kg with no significant adverse effects. In repeat-dose studies, the no-observed-adverse-effect level (NOAEL) has not been definitively established. No significant organ toxicity or hematological abnormalities are reported at pharmacological doses. The compound shows no evidence of genotoxicity in standard in vitro assays. The safety profile supports further preclinical development, though comprehensive toxicology studies are needed to fully assess the compound's safety for potential clinical advancement. The compound is for research use only and is not approved for human use.
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| References | |
| Additional Infomation |
Bark taken from Cephalotaxux hainensis
Harringtonolide is a complex, cage-like norditerpene lactone first isolated from Cephalotaxus harringtonia. It is a potent RACK1 inhibitor (IC50 = 39.66 µM in A375 cells). It inhibits EMT and cell proliferation by affecting the FAK-RACK1 interaction. It is not approved for human use and has not entered clinical trials. It is available as a high-purity research reagent for laboratory use. |
| Molecular Formula |
C19H18O4
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| Molecular Weight |
310.3438
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| Exact Mass |
310.12
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| CAS # |
64761-48-4
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| PubChem CID |
25092280
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| Appearance |
White to off-white solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
605.5±55.0 °C at 760 mmHg
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| Flash Point |
270.5±31.5 °C
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| Vapour Pressure |
0.0±1.7 mmHg at 25°C
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| Index of Refraction |
1.656
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| LogP |
1.53
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
23
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| Complexity |
800
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| Defined Atom Stereocenter Count |
7
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| SMILES |
C[C@@H]1[C@@H]2[C@H]3[C@@H]4[C@@H]5[C@]1(CCC6=CC(=O)C=C(C(=C56)[C@@H]4O2)C)C(=O)O3
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| InChi Key |
QNJIIOHVULPMRL-ZIPZLQRMSA-N
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| InChi Code |
InChI=1S/C19H18O4/c1-7-5-10(20)6-9-3-4-19-8(2)15-17(23-18(19)21)13-14(19)12(9)11(7)16(13)22-15/h5-6,8,13-17H,3-4H2,1-2H3/t8-,13-,14-,15-,16+,17-,19-/m1/s1
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| Chemical Name |
(1S,11S,12R,13R,15R,16R,19S)-8,19-dimethyl-14,17-dioxahexacyclo[13.3.1.01,11.04,10.09,13.012,16]nonadeca-4,7,9-triene-6,18-dione
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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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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) |
DMSO : ~5 mg/mL (~16.11 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 | 3.2223 mL | 16.1114 mL | 32.2227 mL | |
| 5 mM | 0.6445 mL | 3.2223 mL | 6.4445 mL | |
| 10 mM | 0.3222 mL | 1.6111 mL | 3.2223 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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