| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 100mg | |||
| Other Sizes |
| Targets |
Ilexsaponin B1 inhibits tissue factor (tissue thromboplastin, coagulation Factor III). [1]
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|---|---|
| ln Vitro |
In plastic test tubes, Ilexsaponin B1 (final concentrations ≤0.125 mM for whole blood, ≤0.25 mM for PRP and PPP) did not affect the recalcified clotting times of whole blood, platelet-rich plasma (PRP), or platelet-poor plasma (PPP) in the absence of tissue factor. [1]
In the presence of tissue factor (rat lung microsomal fraction), Ilexsaponin B1 prolonged the reduced prothrombin times of whole blood, PRP, and PPP in a dose-dependent manner (concentrations tested up to 0.25 mM). [1] In glass test tubes, the effects of Ilexsaponin B1 on recalcified clotting and prothrombin times were similar to those in plastic tubes, but the anti-tissue factor activity appeared more potent in glass tubes. [1] Ilexsaponin B1 did not affect the recalcified clotting times of whole blood, PRP, or PPP in either plastic or glass tubes without added tissue factor. [1] |
| ln Vivo |
Oral administration of Ilexsaponin B1 (0.3 to 30 mg/kg, twice at a 12 hr interval) prolonged bleeding time in rats. At doses below 0.5 mg/kg, a dose-dependent prolongation was observed; at doses between 0.5 and 30 mg/kg, bleeding time was prolonged approximately two-fold without clear dose-dependency. Compared to aspirin, Ilexsaponin B1 was about ten-fold more potent at the minimum dose producing two-fold prolongation of bleeding time. [1]
Ilexsaponin B1 (0.3 to 10 mg/kg, oral) prolonged the whole blood recalcified clotting time in a dose-dependent manner. At 30 mg/kg, both Ilexsaponin B1 and aspirin tended to shorten the time. [1] Oral administration of Ilexsaponin B1 did not affect the plasma recalcified clotting time. [1] |
| Enzyme Assay |
Tissue factor source: rat lung microsomal fraction. One gram of rat lung was ground for 1 min, mixed with 9 mL saline, then disintegrated in a Teflon homogenizer for 1 min. The mixture was centrifuged at 2000 rpm for 20 min, the sediment discarded, and the supernatant centrifuged at 105,000 × g for 1 hr. The microsomal pellet was suspended in 10 mL saline to make the tissue factor preparation. The undiluted preparation clotted normal rat plasma in 18 sec; a 100-fold dilution gave about 50% acceleration of normal plasma clotting time. [1]
To measure prothrombin time (tissue factor activity), 0.1 mL tissue factor solution and 0.1 mL of 25 mM CaCl₂ were mixed and warmed to 37°C. Then 0.1 mL of PPP or PRP was added, and the time required for clot formation was determined. [1] To test inhibition, Ilexsaponin B1 (3 mM stock solution prepared by dissolving 2.3 mg in 0.03 mL DMSO, then diluted with 50 mM Tris-HCl buffer pH 7.5 to 0.97 mL) was mixed with tissue factor (or saline) and diluted with saline to 0.1 mL total volume. Then 0.1 mL of 25 mM CaCl₂ was added, warmed to 37°C, and 0.1 mL of PPP or PRP was added to determine recalcified clotting time (without tissue factor) or prothrombin time (with tissue factor). [1] |
| Cell Assay |
Preparation of platelet-rich plasma (PRP) and platelet-poor plasma (PPP): Whole blood was collected from rat abdominal aorta into a plastic syringe containing 3.8% sodium citrate (1:10 dilution). PRP was obtained by centrifugation at 200 × g for 30 min at 10°C. PPP was obtained by further centrifugation of the remaining blood at 2000 × g for 30 min at 10°C. [1]
Recalcified clotting time of PRP and PPP: 0.1 mL PRP or PPP and 0.1 mL 0.15 M saline were mixed and warmed to 37°C. Then 0.1 mL of 25 mM CaCl₂ was added, and the time to clot formation was determined. This assay was used to test Ilexsaponin B1 (without tissue factor) in plastic and glass tubes. [1] Prothrombin time (with tissue factor) was measured as described in Enzyme Assay, using PPP or PRP as the plasma source. [1] |
| Animal Protocol |
Male Sprague-Dawley rats (180-200 g for bleeding/clotting time; 200-230 g for blood collection) were used. Animals had free access to food and water. [1]
Oral administration: Ilexsaponin B1 suspended in 1% CMC (carboxymethylcellulose) at doses of 0.3, 0.5, 1, 3, 10, and 30 mg/kg. The compound was administered orally twice with a 12 hr interval (5 mL of 1% CMC suspension per kg body weight each time). Control rats received 1% CMC only. [1] Bleeding time measurement: Two hours after the second administration, rats were anesthetized with intraperitoneal sodium pentobarbital (40 mg/kg). The tail was transected 5 mm from the tip, and the distal 5 cm of the tail was immersed vertically in 0.9% saline at 37.5°C. The time from transection to the moment bleeding stopped was recorded as bleeding time. [1] Blood collection: Four hours after bleeding time measurement, rats were anesthetized with ether, and blood was collected by heart puncture into a plastic tube containing 3.13% sodium citrate (0.1 volume per 0.9 volume blood). [1] Whole blood recalcified clotting time: At room temperature, 1 mL of blood was mixed with 0.2 mL of 1.7% CaCl₂ in a glass tube, and the clotting time was measured. [1] |
| References | |
| Additional Infomation |
It has been reported that holicoside D has been found in Ilex pubescens, and relevant data are available for reference.
Ilexsaponin B1 is reported as the first tissue factor inhibitor from the plant kingdom. [1] Three known protein inhibitors specific for the tissue factor/Factor VIIa complex are mentioned for comparison: lipoprotein-associated coagulation inhibitor (LACI), apolipoprotein A-II, and placental anticoagulant protein (PAP). [1] The study was supported by a grant from Research Center for New Drug Development, The Korean Science and Engineering Foundation. [1] |
| Molecular Formula |
C41H66O13
|
|---|---|
| Molecular Weight |
766.9550
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| Exact Mass |
766.45
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| CAS # |
109008-27-7
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| PubChem CID |
21626431
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| Appearance |
White to off-white solid powder
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| LogP |
2.491
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| Hydrogen Bond Donor Count |
8
|
| Hydrogen Bond Acceptor Count |
13
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| Rotatable Bond Count |
6
|
| Heavy Atom Count |
54
|
| Complexity |
1470
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| Defined Atom Stereocenter Count |
19
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| SMILES |
C[C@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)O[C@H]6[C@@H]([C@H]([C@@H](CO6)O)O)O[C@H]7[C@@H]([C@H]([C@@H]([C@H](O7)CO)O)O)O)C)C)[C@@H]2[C@]1(C)O)C)C(=O)O
|
| InChi Key |
WBDDWIKJZNNKBQ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C41H66O13/c1-20-10-15-41(35(48)49)17-16-38(5)21(32(41)40(20,7)50)8-9-25-37(4)13-12-26(36(2,3)24(37)11-14-39(25,38)6)53-34-31(27(44)22(43)19-51-34)54-33-30(47)29(46)28(45)23(18-42)52-33/h8,20,22-34,42-47,50H,9-19H2,1-7H3,(H,48,49)
|
| Chemical Name |
10-[4,5-dihydroxy-3-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-1-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-2,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-4a-carboxylic acid
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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 : ~100 mg/mL (~130.39 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.26 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 (3.26 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (3.26 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3038 mL | 6.5192 mL | 13.0385 mL | |
| 5 mM | 0.2608 mL | 1.3038 mL | 2.6077 mL | |
| 10 mM | 0.1304 mL | 0.6519 mL | 1.3038 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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