| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 25mg |
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| 100mg | |||
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| Other Sizes |
| ln Vitro |
1. Improvement of endothelial insulin resistance in HUVEC cells (Reference [1]): Kakkalide (0.1-10 μM) reversed insulin resistance induced by high glucose (25 mM) or TNF-α (10 ng/mL) in human umbilical vein endothelial cells (HUVECs). It restored insulin-stimulated phosphorylation of Akt (p-AKT) at Ser473 (by 60% at 10 μM) and endothelial nitric oxide synthase (eNOS) at Ser1177 (by 55% at 10 μM), as measured by Western blot. It also increased nitric oxide (NO) production (detected by DAF-FM DA fluorescence assay) by 40% at 10 μM [1]
2. Suppression of reactive oxygen species (ROS) generation (Reference [1]): Kakkalide (1-10 μM) dose-dependently reduced intracellular ROS levels in HUVECs treated with high glucose or TNF-α. Using DCFH-DA fluorescence staining, ROS levels were decreased by 35% at 5 μM and 50% at 10 μM [1] 3. Inhibition of inflammatory signaling pathways (Reference [1]): Kakkalide (10 μM) suppressed TNF-α-induced nuclear factor-κB (NF-κB) activation in HUVECs. This was demonstrated by reduced nuclear translocation of NF-κB p65 subunit (immunofluorescence) and decreased expression of pro-inflammatory genes (IL-6, MCP-1) via quantitative PCR (mRNA levels reduced by 40%-60%) [1] |
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| ln Vivo |
1. Amelioration of insulin resistance in high-fat diet (HFD)-induced obese mice (Reference [1]): C57BL/6 mice fed a HFD for 12 weeks were treated with Kakkalide (50 mg/kg/day) via oral gavage for 4 weeks. Treatment improved glucose tolerance (glucose tolerance test: AUC reduced by 25%) and insulin sensitivity (insulin tolerance test: AUC reduced by 30%). Plasma insulin levels were decreased by 20%, and hepatic glucose output was suppressed by 30% (determined by pyruvate tolerance test) [1]
2. Reduction of endothelial dysfunction in HFD mice (Reference [1]): Kakkalide treatment restored endothelial-dependent vasodilation in aortic rings isolated from HFD mice. Acetylcholine-induced relaxation was increased from 35% (vehicle) to 60% (Kakkalide). This was associated with increased aortic eNOS phosphorylation (p-eNOS/eNOS ratio increased by 45%) and decreased superoxide production (DHE staining) [1] 3. Anti-inflammatory effects in vivo (Reference [1]): Kakkalide (50 mg/kg/day) reduced plasma levels of pro-inflammatory cytokines (IL-6, TNF-α) by 30%-40% in HFD mice. Liver and adipose tissue sections showed decreased macrophage infiltration (F4/80 immunohistochemistry) and reduced expression of NF-κB target genes [1] |
| Enzyme Assay |
1. ROS measurement using DCFH-DA staining (Reference [1]): HUVECs were loaded with 10 μM DCFH-DA for 30 minutes at 37°C. After treatment with Kakkalide (1-10 μM) and TNF-α (10 ng/mL), cells were washed, and fluorescence intensity (excitation 485 nm, emission 530 nm) was measured using a microplate reader. ROS levels were normalized to cell viability (MTT assay) [1]
2. Nitric oxide (NO) detection using DAF-FM DA (Reference [1]): HUVECs were incubated with 5 μM DAF-FM DA for 30 minutes. After treatment, fluorescence intensity (excitation 495 nm, emission 515 nm) was measured. NO production was calculated relative to sodium nitroprusside (positive control) [1] |
| Cell Assay |
1. Insulin signaling pathway analysis by Western blot (Reference [1]): HUVECs were serum-starved overnight, pretreated with Kakkalide (0.1-10 μM) for 1 hour, and stimulated with insulin (100 nM) for 10 minutes. Cell lysates were prepared, and proteins (p-AKT, AKT, p-eNOS, eNOS, β-actin) were detected by Western blot using specific antibodies. Band intensities were quantified by densitometry [1]
2. NF-κB nuclear translocation assay (Reference [1]): HUVECs were treated with Kakkalide (10 μM) for 1 hour before TNF-α (10 ng/mL) stimulation. Cells were fixed, permeabilized, and stained with anti-NF-κB p65 antibody. Nuclear fluorescence was visualized by confocal microscopy and quantified using ImageJ software [1] 3. Quantitative PCR for inflammatory genes (Reference [1]): Total RNA was extracted from HUVECs using TRIzol reagent. cDNA was synthesized and amplified using SYBR Green PCR master mix. Primers for IL-6, MCP-1, and GAPDH (internal control) were used. Relative gene expression was calculated by the 2^-ΔΔCT method [1] |
| Animal Protocol |
1. High-fat diet-induced obesity model (Reference [1]): C57BL/6 mice (8 weeks old) were fed a HFD (60% kcal from fat) for 12 weeks to induce obesity. Kakkalide (50 mg/kg) was dissolved in 0.5% carboxymethyl cellulose (CMC) and administered orally once daily for 4 weeks. Control mice received vehicle (0.5% CMC). Body weight, food intake, and blood glucose were monitored weekly [1]
2. Glucose and insulin tolerance tests (Reference [1]): After 12 hours of fasting, mice were administered glucose (2 g/kg) or insulin (0.75 U/kg) intraperitoneally. Blood glucose levels were measured at 0, 15, 30, 60, 90, and 120 minutes using a glucometer. Area under the curve (AUC) was calculated using the trapezoidal rule [1] 3. Aortic ring assay for endothelial function (Reference [1]): Aortas were isolated from mice, cleaned of connective tissue, and cut into 2-3 mm rings. Rings were mounted in organ baths with Krebs-Henseleit buffer (37°C, 95% O2/5% CO2). Isometric tension was recorded. After pre-contraction with phenylephrine (1 μM), cumulative concentrations of acetylcholine (10^-9 to 10^-5 M) were added to assess endothelium-dependent relaxation [1] |
| References | |
| Additional Infomation |
It has been reported that cacalide exists in violet (Viola hondoensis) and kudzu (Pueraria montana var. lobata), and there are related data reports.
1. Source and chemical structure: cacalide is a flavonoid glycoside isolated from the seeds of Abrus precatorius (Indian licorice) [1] 2. Mechanism of action: cacalide improves endothelial insulin resistance through the following pathways: (1) inhibiting the generation of reactive oxygen species (ROS), possibly by inhibiting NADPH oxidase; (2) inhibiting NF-κB-mediated inflammation; (3) restoring insulin signaling by enhancing Akt/eNOS phosphorylation [1] 3. Therapeutic potential: cacalide is expected to become an effective drug for treating endothelial dysfunction and insulin resistance associated with obesity and type 2 diabetes [1] |
| Molecular Formula |
C28H32O15
|
|---|---|
| Molecular Weight |
608.5447
|
| Exact Mass |
608.174
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| CAS # |
58274-56-9
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| PubChem CID |
5490351
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| Appearance |
White to off-white solid powder
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| Density |
1.6±0.1 g/cm3
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| Boiling Point |
902.0±65.0 °C at 760 mmHg
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| Flash Point |
296.9±27.8 °C
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| Vapour Pressure |
0.0±0.3 mmHg at 25°C
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| Index of Refraction |
1.693
|
| LogP |
1.62
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| Hydrogen Bond Donor Count |
7
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| Hydrogen Bond Acceptor Count |
15
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| Rotatable Bond Count |
8
|
| Heavy Atom Count |
43
|
| Complexity |
972
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| Defined Atom Stereocenter Count |
9
|
| SMILES |
COC1=CC=C(C=C1)C2=COC3=CC(=C(C(=C3C2=O)O)OC)O[C@H]4[C@@H]([C@H]([C@@H]([C@H](O4)CO[C@H]5[C@@H]([C@H]([C@@H](CO5)O)O)O)O)O)O
|
| InChi Key |
QTVAYNGFFDZGDR-CIJVEFAYSA-N
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| InChi Code |
InChI=1S/C28H32O15/c1-37-12-5-3-11(4-6-12)13-8-39-15-7-16(26(38-2)22(33)18(15)19(13)30)42-28-25(36)23(34)21(32)17(43-28)10-41-27-24(35)20(31)14(29)9-40-27/h3-8,14,17,20-21,23-25,27-29,31-36H,9-10H2,1-2H3/t14-,17-,20+,21-,23+,24-,25-,27+,28-/m1/s1
|
| Chemical Name |
5-hydroxy-6-methoxy-3-(4-methoxyphenyl)-7-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-[[(2S,3R,4S,5R)-3,4,5-trihydroxyoxan-2-yl]oxymethyl]oxan-2-yl]oxychromen-4-one
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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: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 : ~50 mg/mL (~82.16 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 | 1.6433 mL | 8.2164 mL | 16.4328 mL | |
| 5 mM | 0.3287 mL | 1.6433 mL | 3.2866 mL | |
| 10 mM | 0.1643 mL | 0.8216 mL | 1.6433 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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