| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 25mg | |||
| 50mg | |||
| Other Sizes |
| ln Vitro |
Galgravin exhibits cytotoxic activity with an IC50 of 16.5 µg/mL against tumor cells derived from human leukemia[2].
Galgravin (compound 8) displayed cytotoxic activity against human leukemia HL-60 cells with an IC50 value of 16.5 ± 0.8 μg/mL. It also showed activity against murine melanoma B16F10 cells (IC50 = 60.0 ± 3.2 μg/mL) and human cervical carcinoma HeLa cells (IC50 = 68.0 ± 3.2 μg/mL), but was inactive (IC50 > 100 μg/mL) against human breast adenocarcinoma MCF7 and human melanoma A2058 cell lines [2]. In HL-60 cells treated with 50 μg/mL of Galgravin for 24 h, morphological changes including cytoplasm retraction, plasma membrane bleb formation (apoptotic bodies), and condensation of nuclear material (pyknosis) followed by chromatin fragmentation (karyorrhexis) were observed by light and fluorescence microscopy. Chromatin condensation was observed in 37.0% of total cells stained with DAPI. DNA fragmentation analysis by agarose gel electrophoresis revealed a ladder pattern, indicating oligonucleosomal fragmentation. Phosphatidylserine externalization was detected by Annexin V/PI assay: at 100 μg/mL for 24 h, early apoptotic cells (AV+/PI-) accounted for 12.7% and late apoptotic cells (AV+/PI+) for 1%, compared to negative control. Loss of mitochondrial membrane potential (ΔΨm) was measured using TMRE dye: 52.2% of HL-60 cells exhibited low ΔΨm after treatment with 100 μg/mL Galgravin for 24 h. Western blot analysis showed increased cleavage of PARP (89 kDa fragment) in HL-60 cells treated with 100 μg/mL Galgravin for 24 h [2]. |
|---|---|
| ln Vivo |
In the acetic acid-induced abdominal constriction test in male Swiss albino mice (20-25 g), oral administration of Galgravin (compound 2) at doses of 5, 10, and 20 mg/kg produced dose-dependent inhibition of constrictions. The number of constrictions was 9.67 ± 3.3 (55.0% inhibition, P<0.05) at 5 mg/kg, 8.50 ± 1.4 (60.5% inhibition, P<0.05) at 10 mg/kg, and 8.50 ± 2.1 (60.5% inhibition, P<0.05) at 20 mg/kg, compared to control (21.5 ± 4.2) [1].
In the carrageenan-induced paw oedema test in male Wistar rats (150-180 g), oral administration of Galgravin (compound 2) at 10 mg/kg and 20 mg/kg significantly inhibited oedema formation by 52.7% and 71.4%, respectively, measured at the third hour after carrageenan injection. At 5 mg/kg, no significant effect was reported [1]. In the hot plate test in rats (55 ± 1°C), Galgravin (compound 2) at 20 mg/kg (p.o.) was inactive, showing no significant increase in response latency compared to control [1]. |
| Cell Assay |
Cytotoxicity was assessed using the MTT-based colorimetric assay. Cells (1 × 10^4 per well) were seeded in 96-well plates and incubated with different concentrations of Galgravin (0 to 100 μg/mL) for 24 h at 37°C in 5% CO2. Then MTT solution was added, and formazan crystals were dissolved. Absorbance was read at 570 nm with reference at 650 nm. IC50 values were calculated from correlation curves [2].
For morphology analysis, HL-60 cells (1 × 10^4) were seeded in 96-well plates and treated with 50 μg/mL Galgravin for 24 h. Images were captured using a light inverted microscope at 400× magnification. For chromatin condensation analysis, cells were harvested, washed with PBS, fixed with methanol for 15 min at room temperature, stained with 10 μg/mL DAPI in PBS for 15 min, and examined under an inverted fluorescence microscope (40× objective) [2]. DNA fragmentation was assessed by agarose gel electrophoresis. HL-60 cells (1 × 10^5) were incubated with 50 μg/mL Galgravin for 24 h. Total DNA was extracted, processed, and analyzed on a 1% agarose gel [2]. Phosphatidylserine externalization was detected using Annexin V-FITC Apoptosis kit. HL-60 cells (5 × 10^5) were cultured in 6-well plates and incubated with 100 μg/mL Galgravin for 24 h. After washing with PBS, cells were resuspended in binding buffer containing Annexin V-FITC and propidium iodide (PI) for 10 min at room temperature, then analyzed by flow cytometry [2]. Mitochondrial membrane potential (ΔΨm) was measured using the cationic lipophilic dye tetramethylrhodamine ethyl ester (TMRE). HL-60 cells (5 × 10^5) were incubated with 100 μg/mL Galgravin for 24 h, then processed and analyzed by flow cytometry [2]. For Western blotting, HL-60 cells (5 × 10^5) treated with 100 μg/mL Galgravin for 24 h were lysed in SDS sample buffer with phosphatase and protease inhibitors. Total proteins were separated by SDS-PAGE and transferred to membranes. Antibodies against cleaved PARP and β-actin (loading control) were used, followed by HRP-conjugated secondary antibody. Immunoreactivity was detected using chemiluminescence [2]. |
| Animal Protocol |
For the acetic acid-induced abdominal constriction test, male Swiss albino mice (20-25 g) were used. Galgravin (compound 2) was administered orally at doses of 5, 10, or 20 mg/kg in a vehicle of 5% Tween-saline solution. Thirty minutes after administration, 0.6% acetic acid (0.1 mL/10 g body weight) was injected intraperitoneally. The number of abdominal constrictions was counted for 20 min after acetic acid injection. Indometacin (10 mg/kg, p.o.) was used as reference drug. Control animals received vehicle only (0.2 mL/animal, p.o.) [1].
For the carrageenan-induced paw oedema test, male Wistar rats (150-180 g) were used. Galgravin (compound 2) was administered orally at doses of 5, 10, or 20 mg/kg (in 5% Tween-saline solution) 30 min before injection of 0.1 mL carrageenan (100 μg) into the plantar surface of the rat paw. Paw volume was measured using a plethysmometer at 1-h intervals for 5 h, and activity was assessed at the third hour when maximum oedema occurred. Indometacin (10 mg/kg, p.o.) was used as positive control. Negative control group received vehicle (5% Tween-saline solution) [1]. For the hot plate test, male Wistar rats (150-180 g) were used. The hot plate was maintained at 55 ± 1°C. Animals were selected 24 h before the experiment based on reactivity (response latency within 6 s). Galgravin (compound 2) was administered orally at 20 mg/kg (in 5% Tween-saline solution). Response latency (time to shaking or licking of paws or jumping) was recorded 60 min after administration, with a cut-off of 30 s. Morphine (4.0 mg/kg, s.c.) was used as positive control. Results were expressed as percent of maximum possible effect (% MPE) [1]. |
| References |
|
| Additional Infomation |
Galgravin belongs to the aryltetrahydrofuran class of compounds, with two 3,4-dimethoxyphenyl substituents at positions 2 and 5, and two methyl groups at positions 3 and 4. It possesses functions including maintaining bone density, neuroprotection, inhibiting platelet aggregation, and acting as a plant metabolite. It is an aryltetrahydrofuran, dimethoxyphenyl, cyclic compound, and lignan. Galgravin has been reported in Schisandra chinensis, Schisandra propinqua, and other organisms with relevant data. See also: Galbelgin (note moved here).
In the abdominal constriction test, Galgravin showed peripheral analgesic activity as it was inactive in the hot plate test (which is selective for opioid-like compounds). The mechanism may be related to inhibition of platelet-activating factor (PAF) receptors and/or other inflammatory mediators such as prostaglandins and kinins, based on previous reports on tetrahydrofuran lignans [1]. Galgravin (compound 8) induced apoptosis in HL-60 cells via the intrinsic mitochondrial pathway, as evidenced by loss of mitochondrial membrane potential, PARP cleavage, and characteristic apoptotic hallmarks such as blebbing, chromatin condensation, and phosphatidylserine externalization [2]. |
| Molecular Formula |
C22H28O5
|
|---|---|
| Molecular Weight |
372.4547
|
| Exact Mass |
372.194
|
| Elemental Analysis |
C, 70.94; H, 7.58; O, 21.48
|
| CAS # |
528-63-2
|
| PubChem CID |
101749
|
| Appearance |
Solid powder
|
| LogP |
4.805
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
27
|
| Complexity |
418
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
O1[C@]([H])(C2C([H])=C([H])C(=C(C=2[H])OC([H])([H])[H])OC([H])([H])[H])[C@@]([H])(C([H])([H])[H])[C@@]([H])(C([H])([H])[H])[C@]1([H])C1C([H])=C([H])C(=C(C=1[H])OC([H])([H])[H])OC([H])([H])[H]
|
| InChi Key |
JLJAVUZBHSLLJL-DQEHQXCCSA-N
|
| InChi Code |
InChI=1S/C22H28O5/c1-13-14(2)22(16-8-10-18(24-4)20(12-16)26-6)27-21(13)15-7-9-17(23-3)19(11-15)25-5/h7-14,21-22H,1-6H3/t13-,14+,21-,22+
|
| Chemical Name |
(2S,3S,4R,5R)-2,5-bis(3,4-dimethoxyphenyl)-3,4-dimethyloxolane
|
| Synonyms |
(±)-Galgravin; Galgravin
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 2.6849 mL | 13.4246 mL | 26.8492 mL | |
| 5 mM | 0.5370 mL | 2.6849 mL | 5.3698 mL | |
| 10 mM | 0.2685 mL | 1.3425 mL | 2.6849 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
