| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Natural product; Akt/mTOR
|
|---|---|
| ln Vitro |
In N2a and SH-SY5Y cells, corynoxine (6.25-25 μM; 6–12 h) dose-dependently enhances the expression of LC3-II, an autophagy-specific marker[1]. Using autophagy induction, corynoxine (25 μM; 48 h) stimulates the degradation of wild type (WT) and mutant (A53T) α-syn in inducible PC12 cells[1].
Corynoxine (10, 20, 40 μM) treatment for 24 hours significantly reduced the level of A53T α-synuclein in a stable PC12 cell model in a dose-dependent manner, as detected by Western blot analysis. [1] Corynoxine (20 μM) treatment for 24 hours increased the protein levels of the autophagy marker LC3-II and decreased the level of the autophagy substrate p62 in PC12 cells expressing A53T α-synuclein, indicating the induction of autophagy. [1] The autophagy-enhancing effect of Corynoxine was confirmed by the observation of increased numbers of GFP-LC3 puncta (autophagosomes) in HeLa cells transfected with a GFP-LC3 plasmid after treatment with 20 μM Corynoxine for 24 hours. [1] |
| ln Vivo |
Mice that are given corynoxine (100–100 mg/kg via oral gavage) show extended thiopental-induced hypnosis[2]. In drosophila, corynoxine (10–100 μM) promotes autophagy over 12 hours[1].
Cory (corynoxine) Induces Autophagy in Drosophila: To confirm the autophagy-inducing effect of Cory in vivo, we firstly crossed Cg-GAL4 fly lines to UAS-GFP-Atg8a fly lines at 25 °C, and got the 2nd instar larvae of Cg-GAL4 > UAS-GFP-Atg8a. We fed these larvae with food containing different concentrations of Cory (10-100 μM) for 12 h at 25 °C. In our previous study, Cory B was confirmed to induce autophagy in Drosophila; therefore we chose it as a positive control and using 0.4 % DMSO as a negative control. We collected the treated larvae and prepared the fat bodies for photographing through a confocal microscope. Atg8 (Autophagy-related protein), the homolog of LC3, is an ubiquitin-like protein required for the formation of autophagosomal membranes. Compared with the control, both Cory (10-100 μM) and Cory B (100 μM) induced the puncta formation of Atg8 (Fig. 3), suggesting that Cory could also induce autophagy in the fat bodies of Drosophila larvae [1]. |
| Cell Assay |
Western Blot Analysis[1]
Cell Types: N2a and SH-SY5Y cells Tested Concentrations: 6.25, 12.5, 25 μM Incubation Duration: 6, 12 hrs (hours) Experimental Results: Induced autophagy in neuronal cell lines. Cell culture and treatment: PC12 cells stably expressing human A53T mutant α-synuclein were used. Cells were cultured in growth medium. For treatment, Corynoxine was dissolved in DMSO and then diluted in the cell culture medium to the final concentrations (10, 20, 40 μM). An equal volume of DMSO was used as the vehicle control. Cells were treated for 24 hours. [1] Western blot analysis: After treatment, cells were harvested and lysed. The protein concentration of the lysates was determined. Equal amounts of protein were separated by SDS-PAGE and then transferred to a membrane. The membrane was blocked and then incubated with primary antibodies against α-synuclein, LC3, p62, phospho-Akt (Ser473), Akt, phospho-mTOR (Ser2448), mTOR, and β-actin (as a loading control) overnight at 4°C. After washing, the membrane was incubated with a secondary antibody. Protein bands were visualized using an enhanced chemiluminescence detection system. [1] GFP-LC3 puncta formation assay: HeLa cells were transfected with a GFP-LC3 plasmid. After 24 hours, the cells were treated with 20 μM Corynoxine or vehicle for another 24 hours. The cells were then fixed. The formation of GFP-LC3 puncta (autophagosomes) was observed and imaged using a fluorescence microscope. Cells with more than 10 GFP-LC3 puncta were considered to have undergone autophagy. [1] |
| Animal Protocol |
Hypnosis test: The alkaloid fraction from Uncaria macrophylla (containing Corynoxine, Corynoxine B, Rhynchophylline, and Isorhynchophylline) was administered intraperitoneally (i.p.) to mice. The test was conducted by injecting thiopental sodium (40 mg/kg, i.p.) 30 minutes after the administration of the alkaloid fraction. The loss of the righting reflex was used as the criterion for hypnosis. The duration of hypnosis (sleeping time) was recorded. The alkaloid fraction was dissolved in a saline solution containing a drop of Tween 80. [2]
|
| Toxicity/Toxicokinetics |
No specific toxicity data (e.g., LD50, organ toxicity) for purified Corynoxine is reported in the provided literature. [2] only states that the alkaloid fraction from Uncaria macrophylla (which includes Corynoxine) did not cause any observable toxic symptoms or mortality in mice at the tested doses (5, 10, 20 mg/kg, i.p.). [2]
|
| References | |
| Additional Infomation |
Corynoxine is a natural oxindole alkaloid isolated from the hooks of Uncaria macrophylla. [1]
The study demonstrates that Corynoxine promotes the clearance of alpha-synuclein, a protein implicated in Parkinson's disease, by enhancing autophagy. [1] The mechanism of action involves the inhibition of the Akt/mTOR pathway. Corynoxine (20 μM) treatment decreased the phosphorylation levels of Akt (at Ser473) and mTOR (at Ser2448) in PC12 cells expressing A53T α-synuclein, as shown by Western blot, without affecting the total protein levels of Akt and mTOR. [1] The hypnotic effects reported in [2] are for a mixture of four alkaloids (Corynoxine, Corynoxine B, Rhynchophylline, Isorhynchophylline), not for Corynoxine alone. The mixture (20 mg/kg, i.p.) significantly prolonged thiopental-induced sleeping time in mice. The effect of the mixture was stronger than that of any single alkaloid administered at an equivalent dose. [2] |
| Molecular Formula |
C22H29CLN2O4
|
|---|---|
| Molecular Weight |
420.93
|
| Exact Mass |
420.181585
|
| Related CAS # |
Corynoxine;6877-32-3
|
| PubChem CID |
21148450
|
| Appearance |
Typically exists as solid at room temperature
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
29
|
| Complexity |
663
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
CC[C@@H]1CN2CC[C@@]3([C@@H]2C[C@@H]1/C(=C\OC)/C(=O)OC)C4=CC=CC=C4NC3=O.Cl
|
| InChi Key |
YZEHFWIJWUTTOF-UXWWZSLJSA-N
|
| InChi Code |
InChI=1S/C22H28N2O4.ClH/c1-4-14-12-24-10-9-22(17-7-5-6-8-18(17)23-21(22)26)19(24)11-15(14)16(13-27-2)20(25)28-3;/h5-8,13-15,19H,4,9-12H2,1-3H3,(H,23,26);1H/b16-13+;/t14-,15+,19+,22+;/m1./s1
|
| Chemical Name |
methyl (E)-2-[(3S,6'S,7'S,8'aS)-6'-ethyl-2-oxospiro[1H-indole-3,1'-3,5,6,7,8,8a-hexahydro-2H-indolizine]-7'-yl]-3-methoxyprop-2-enoate;hydrochloride
|
| Synonyms |
Corynoxine (hydrochloride); orb1689909;
|
| 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 | 2.3757 mL | 11.8785 mL | 23.7569 mL | |
| 5 mM | 0.4751 mL | 2.3757 mL | 4.7514 mL | |
| 10 mM | 0.2376 mL | 1.1878 mL | 2.3757 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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