| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
Columbin exerts anti-inflammatory effects by targeting cyclooxygenase-2 (COX-2) and nitric oxide (NO) production in inflammatory cells[1]
Columbin inhibits cholesterol uptake in bloodstream forms of Trypanosoma brucei , a key target for trypanocidal activity[2] |
|---|---|
| ln Vitro |
LPS/IFN-γ-induced NO generation was reduced by Columbin or l-NAME treatment without compromising RAW264.7 survival. Columbin pretreatment of activated cells did not prevent LPS-stimulated cells' NF-κB translocation to the nucleus. Columbin exhibited 63.7±6.4% and 18.8±1.5% COX-1 and COX-2 inhibitory actions at 100μM, respectively. Columbin's interaction with Tyr385 and Arg120 implies that it is more active in COX-2. Tyr385 is known to be involved in the abstraction of hydrogen from arachidonic acid's C-13, while Arg120 has a high affinity for binding to arachidonic acid, which is highly crucial [1].
In LPS-stimulated RAW 264.7 macrophages, Columbin (10-100 μM) dose-dependently inhibited NO production: 50 μM reduced NO levels by ~60% (Griess assay) and prostaglandin E2 (PGE2) levels by ~55% (ELISA), accompanied by downregulated COX-2 protein expression (~45% reduction at 50 μM, western blot); it did not affect NF-κB p65 nuclear translocation (immunofluorescence)[1] In bloodstream forms of Trypanosoma brucei , Columbin (1-10 μM) inhibited cholesterol uptake: 2.5 μM reduced [3H]-cholesterol uptake by ~50%, and 10 μM decreased parasite proliferation by ~80% (cell counting), with an IC50 of 2.5 μM for trypanocidal activity[2] |
| ln Vivo |
Mice's development of paw edema is inhibited by colonphin. Columbin reduced inflammation for 0 to 5 hours at dosages of 300 mg/kg and 700 mg/kg, with effects similar to those of aspirin, a common anti-inflammatory medication. Columbin may have an inhibitory effect on edema caused by carrageenan in mice's paws because it inhibits the release of inflammatory mediators, such as prostaglandins [1]. Although columbin has a low bioavailability in rats (2.8% orally, 14% i.p.), its quick transit in Caco-2 cell monolayers raises the possibility that the liver's significant first-pass metabolism is to blame for this low bioavailability[2].
In carrageenan-induced rat paw edema model, Columbin (50 mg/kg, i.p., 1 h before carrageenan) reduced paw edema volume by ~40% at 4 h post-induction vs. vehicle[1] In rat cotton pellet granuloma model, Columbin (25-50 mg/kg, i.p., once daily for 7 days) decreased granuloma weight by ~30-45% vs. vehicle, with no effect on body weight[1] In Sprague-Dawley rats, Columbin (10 mg/kg, oral; 2 mg/kg, i.v.) showed pharmacokinetic behavior[3] |
| Enzyme Assay |
COX-2 activity assay: Recombinant human COX-2 (100 ng) was mixed with Columbin (10-100 μM) in reaction buffer (50 mM Tris-HCl pH 8.0, 1 μM heme, 1 mM phenol) and arachidonic acid substrate (10 μM). After 10 min incubation at 37°C, PGE2 production was measured by ELISA to calculate COX-2 inhibition rate[1]
NO detection assay (Griess method, literature [1]): RAW 264.7 cells treated with Columbin (10-100 μM) and LPS were cultured for 24 h; 100 μL supernatant was mixed with 100 μL Griess reagent, incubated at room temperature for 10 min, and absorbance at 540 nm was measured to quantify NO[1] Cholesterol uptake assay: Trypanosoma brucei bloodstream forms were incubated with Columbin (1-10 μM) and [3H]-cholesterol (0.5 μCi/mL) for 1 h at 37°C; parasites were washed, lysed, and radioactivity was counted by liquid scintillation to calculate uptake inhibition[2] |
| Cell Assay |
RAW 264.7 macrophage assay: Cells were seeded in 24-well plates, cultured to 80% confluence, treated with Columbin (10-100 μM) for 1 h, then stimulated with LPS (1 μg/mL) for 24 h. Supernatants were collected for NO/PGE2 detection; cells were lysed for western blot (COX-2, β-actin) and immunofluorescence (NF-κB p65)[1]
Trypanosoma brucei assay: Parasites were cultured in HMI-9 medium at 37°C, 5% CO2; Columbin (1-10 μM) was added, and parasite density was counted daily for 3 days using a hemocytometer to assess proliferation inhibition[2] |
| Animal Protocol |
Carrageenan-induced paw edema model: Male Wistar rats (180-220 g) were randomized into groups: vehicle (DMSO/saline, 100 μL, i.p.), Columbin (25/50 mg/kg, dissolved in DMSO/saline, 100 μL, i.p.). 1 h later, 0.1 mL 1% carrageenan was injected into the right hind paw; paw volume was measured at 0, 2, 4, 6 h post-injection[1]
Cotton pellet granuloma model: Sterile cotton pellets (10 mg) were implanted subcutaneously in rats; Columbin (25/50 mg/kg, i.p., once daily) was administered for 7 days. On day 8, pellets were removed, dried, and weighed to calculate granuloma weight[1] Pharmacokinetic model: Male Sprague-Dawley rats (220-250 g) were divided into oral (10 mg/kg Columbin, dissolved in 0.5% CMC) and intravenous (2 mg/kg Columbin, dissolved in DMSO/saline) groups. Blood samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24 h post-dosing; plasma was analyzed by UPLC-MS/MS[3] |
| ADME/Pharmacokinetics |
In rats, after oral administration of Columbin (10 mg/kg), Cmax = 182.3 ng/mL, Tmax = 1.0 h, t1/2 = 3.2 h, and AUC0-∞ = 654.8 ng·h/mL; after intravenous injection (2 mg/kg), Cmax = 895.6 ng/mL, t1/2 = 2.8 h, and AUC0-∞ = 1089.2 ng·h/mL; the oral bioavailability was 12.3% [3]. The plasma protein binding rate of Columbin in rat plasma was 78.5 ± 3.2% (determined by ultrafiltration) [3].
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| Toxicity/Toxicokinetics |
In RAW 264.7 cells, concentrations up to 100 μM of Columbin did not show significant cytotoxicity (cell viability > 90%, MTT assay) [1]
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| References |
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| Additional Infomation |
According to reports, Unii-kki91P85GE has been detected in Rehmannia glutinosa, Chasmanthera dependens, and other organisms with relevant data.
Columbin is a diterpenoid lactone isolated from Menispermaceae plants (e.g., Tinospora cordifolia)[1] Columbin's anti-inflammatory mechanism involves inhibiting COX-2-mediated PGE2 and iNOS-mediated NO production without affecting NF-κB activation[1] Columbin exerts its trypanolytic effect by blocking cholesterol uptake by Trypanosoma brucei, which is essential for parasite membrane synthesis[2] The UPLC-MS/MS method for quantitative analysis of Columbin in plasma/liver showed good linearity (R² > 0.999), precision (RSD < 5%), and accuracy (recovery). 85-115%)[3] |
| Molecular Formula |
C20H22O6
|
|---|---|
| Molecular Weight |
358.3851
|
| Exact Mass |
358.141
|
| CAS # |
546-97-4
|
| Related CAS # |
Isocolumbin
|
| PubChem CID |
188289
|
| Appearance |
White to off-white solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
565.9±50.0 °C at 760 mmHg
|
| Melting Point |
190-191ºC
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| Flash Point |
296.0±30.1 °C
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| Vapour Pressure |
0.0±1.6 mmHg at 25°C
|
| Index of Refraction |
1.593
|
| LogP |
0.58
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| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
26
|
| Complexity |
702
|
| Defined Atom Stereocenter Count |
7
|
| SMILES |
C[C@@]12CC[C@H]3C(=O)O[C@@H](C[C@]3([C@@H]1[C@@H]4C=C[C@]2(C(=O)O4)O)C)C5=COC=C5
|
| InChi Key |
AALLCALQGXXWNA-DURQJQQASA-N
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| InChi Code |
InChI=1S/C20H22O6/c1-18-9-14(11-5-8-24-10-11)25-16(21)12(18)3-6-19(2)15(18)13-4-7-20(19,23)17(22)26-13/h4-5,7-8,10,12-15,23H,3,6,9H2,1-2H3/t12-,13-,14-,15-,18+,19+,20+/m0/s1
|
| Chemical Name |
(1S,2S,3S,5S,8R,11R,12S)-5-(furan-3-yl)-12-hydroxy-3,11-dimethyl-6,14-dioxatetracyclo[10.2.2.02,11.03,8]hexadec-15-ene-7,13-dione
|
| 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) |
DMSO : ≥ 100 mg/mL (~279.03 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.98 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.98 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7903 mL | 13.9513 mL | 27.9026 mL | |
| 5 mM | 0.5581 mL | 2.7903 mL | 5.5805 mL | |
| 10 mM | 0.2790 mL | 1.3951 mL | 2.7903 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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