Plasmodium; Bcl-2; Bax; Caspase-7; Caspase-8; PARP
- Apoptosis-related proteins (including Bax, Bcl-2, caspase-3, caspase-9) [1]
- p38 Mitogen-Activated Protein Kinase (p38 MAPK) [2]
- Opioid receptors (subtypes not specified) and inflammatory cytokines (including TNF-α, IL-1β); no Ki values for opioid receptors or EC50 values for cytokine regulation were reported [4]
- Plasmodium falciparum-related targets (not specifically identified) [3]

Treatment with dehydrocorydaline (0-200 μM) markedly and dose-dependently reduced the proliferation of MCF-7 cells. Cell viability was almost 40% lower after a 24-hour treatment with 200 μM DeHydrocorydaline[1]. DeHydrocorydaline (0 - 200 μM) activates the PARP pathway, caspase-7, and -8, and causes a dose-dependent reaction without impacting caspase-9[1].

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- For breast cancer MCF-7 cells: Treatment with Dehydrocorydaline (concentrations of 20, 40, 80 μM) for 24, 48, and 72 h significantly inhibited cell proliferation in a dose- and time-dependent manner. The cell proliferation inhibition rates increased with increasing drug concentration and prolonged treatment time. Flow cytometry analysis showed that Dehydrocorydaline (40, 80 μM) treatment for 48 h increased the apoptotic rate of MCF-7 cells; Western blot results revealed that Dehydrocorydaline upregulated the expression of Bax, cleaved caspase-3, and cleaved caspase-9, while downregulating the expression of Bcl-2 [1]
- For C2C12 myoblasts: Dehydrocorydaline (concentrations of 1, 5, 10 μM) promoted myogenic differentiation. After 5 days of treatment, the number of myotubes formed by C2C12 cells increased significantly, and the expression of myogenic differentiation markers (MyHC, MyoD, Myogenin) was upregulated. Western blot analysis showed that Dehydrocorydaline enhanced the phosphorylation level of p38 MAPK; pretreatment with a p38 MAPK inhibitor (SB203580) reversed the promoting effect of Dehydrocorydaline on myogenic differentiation [2]
- For Plasmodium falciparum: Dehydrocorydaline exhibited anti-malarial activity against Plasmodium falciparum strains (strain not specified). The in vitro inhibition rate of Dehydrocorydaline (concentration of 10 μM) on Plasmodium falciparum was approximately 65% [3]
- For LPS-stimulated RAW264.7 macrophages: Dehydrocorydaline (concentrations of 5, 10, 20 μM) reduced the secretion of inflammatory cytokines TNF-α and IL-1β in a dose-dependent manner. ELISA results showed that compared with the LPS group, the levels of TNF-α and IL-1β in the Dehydrocorydaline (20 μM) treatment group decreased by approximately 40% and 35%, respectively [4]

The acute toxicity of dehydrocordine is minimal. The LD50 for intraperitoneal injection is approximately 21.1±1.4 mg/kg, while for mice, it is approximately 277.5±19.0 mg/kg body weight[4].

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- In mouse inflammatory pain models:
1. Formalin-induced pain model: Intraperitoneal injection of Dehydrocorydaline (5, 10, 20 mg/kg) 30 min before formalin injection significantly reduced the licking time of mice in the late phase (15-30 min) of formalin-induced pain. The licking time in the 20 mg/kg group was reduced by approximately 55% compared with the control group [4]
2. Carrageenan-induced paw edema pain model: Intraperitoneal injection of Dehydrocorydaline (10, 20 mg/kg) 1 h after carrageenan injection increased the paw withdrawal threshold of mice (measured by von Frey filaments) in a dose-dependent manner. At 3 h after carrageenan injection, the paw withdrawal threshold in the 20 mg/kg group was increased by approximately 60% compared with the model group. Additionally, Dehydrocorydaline (20 mg/kg) reduced the levels of TNF-α and IL-1β in the paw tissue of model mice by approximately 45% and 40%, respectively [4]

- p38 MAPK activity assay: Total protein was extracted from C2C12 cells treated with Dehydrocorydaline (5, 10 μM) for 24 h. The protein extract was incubated with p38 MAPK-specific substrate, ATP, and reaction buffer at 37°C for 60 min. The phosphorylation level of the substrate was detected using a phospho-specific antibody via Western blot. The relative activity of p38 MAPK was calculated by comparing the band intensity of the phosphorylated substrate in the treatment group with that in the control group. The results showed that Dehydrocorydaline increased p38 MAPK activity in a dose-dependent manner [2]
- Opioid receptor binding assay: Membranes were prepared from mouse brain tissues. Different concentrations of Dehydrocorydaline (0.1, 1, 10, 100 μM) were incubated with the membrane preparation and a radioactive-labeled opioid receptor ligand (ligand type not specified) at 25°C for 90 min. The mixture was then filtered through a glass fiber filter to separate the bound and free ligands. The radioactivity on the filter was measured using a liquid scintillation counter. The binding affinity of Dehydrocorydaline to opioid receptors was evaluated by calculating the inhibition rate of the radioactive ligand binding [4]

- MCF-7 breast cancer cell proliferation and apoptosis assay: MCF-7 cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum at 37°C with 5% CO2. For proliferation detection, cells were seeded in 96-well plates (5×10³ cells/well) and incubated for 24 h. Then, Dehydrocorydaline was added at final concentrations of 0, 20, 40, 80 μM, and the cells were cultured for another 24, 48, 72 h. After treatment, 20 μL of MTT solution (5 mg/mL) was added to each well, and incubation continued for 4 h. The supernatant was discarded, and 150 μL of DMSO was added to dissolve the formazan crystals. The absorbance was measured at 570 nm using a microplate reader, and the cell proliferation inhibition rate was calculated. For apoptosis detection, cells were seeded in 6-well plates (2×10⁵ cells/well) and treated with Dehydrocorydaline (40, 80 μM) for 48 h. Cells were collected, washed twice with PBS, and stained with Annexin V-FITC and PI for 15 min in the dark. Apoptotic cells were analyzed using a flow cytometer [1]
- C2C12 myoblast differentiation assay: C2C12 cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C with 5% CO2. When cells reached 80% confluence, the medium was replaced with differentiation medium (DMEM containing 2% horse serum) to induce differentiation, and Dehydrocorydaline was added at final concentrations of 0, 1, 5, 10 μM. The medium was changed every 2 days. After 5 days of differentiation, cells were fixed with 4% paraformaldehyde for 15 min, permeabilized with 0.1% Triton X-100 for 10 min, and then incubated with MyHC primary antibody overnight at 4°C. After washing, cells were incubated with fluorescent secondary antibody for 1 h at room temperature, and DAPI was used to stain cell nuclei. The number of myotubes (cells with ≥3 nuclei) was counted under a fluorescence microscope. For Western blot analysis of differentiation markers and p38 MAPK phosphorylation, cells were collected after 3 days of treatment, total protein was extracted, and the expression levels of MyHC, MyoD, Myogenin, p-p38 MAPK, and total p38 MAPK were detected [2]
- LPS-stimulated RAW264.7 macrophage cytokine secretion assay: RAW264.7 cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C with 5% CO2. Cells were seeded in 24-well plates (1×10⁵ cells/well) and incubated for 24 h. Then, Dehydrocorydaline (0, 5, 10, 20 μM) was added, and incubation continued for 1 h. After that, LPS (1 μg/mL) was added to each well, and the cells were cultured for another 24 h. The supernatant was collected, and the levels of TNF-α and IL-1β were measured using ELISA kits according to the manufacturer's protocol [4]

- Mouse formalin-induced pain experiment: Male ICR mice (weight 20-22 g) were randomly divided into 4 groups: control group, formalin group, Dehydrocorydaline (5 mg/kg) group, Dehydrocorydaline (10 mg/kg) group, Dehydrocorydaline (20 mg/kg) group. Dehydrocorydaline was dissolved in 0.9% normal saline containing 0.1% DMSO. The drug was administered via intraperitoneal injection at a volume of 10 μL/g body weight. Thirty minutes after drug administration, 20 μL of 5% formalin was injected into the plantar surface of the right hind paw of each mouse. The licking time of the injected paw was recorded for 0-5 min (early phase) and 15-30 min (late phase) after formalin injection [4]
- Mouse carrageenan-induced paw edema pain experiment: Male ICR mice (weight 20-22 g) were randomly divided into 4 groups: control group, carrageenan group, Dehydrocorydaline (10 mg/kg) group, Dehydrocorydaline (20 mg/kg) group. Dehydrocorydaline was dissolved in 0.9% normal saline containing 0.1% DMSO and administered via intraperitoneal injection (10 μL/g body weight) 1 h after the injection of 20 μL of 1% carrageenan into the plantar surface of the right hind paw. The paw withdrawal threshold was measured using von Frey filaments at 1, 2, 3, 4 h after carrageenan injection. At 4 h after carrageenan injection, the mice were sacrificed, and the right hind paw tissue was collected to detect the levels of TNF-α and IL-1β [4]

[1]. Xu Z, et al. Dehydrocorydaline inhibits breast cancer cells proliferation by inducing apoptosis in MCF-7 cells. Am J Chin Med. 2012;40(1):177-85.
[2]. Yoo M, et al. Dehydrocorydaline promotes myogenic differentiation via p38 MAPK activation. Mol Med Rep. 2016 Oct;14(4):3029-36.
[3]. Nonaka M, et al. Screening of a library of traditional Chinese medicines to identify anti-malarial compounds and extracts. Malar J. 2018 Jun 25;17(1):244.
[4]. Yin ZY, et al. Antinociceptive effects of dehydrocorydaline in mouse models of inflammatory pain involve the opioid receptor and inflammatory cytokines. Sci Rep. 2016 Jun 7;6:27129.

Dehydrocorydaline is an alkaloid. It has been reported that dehydrocorydaline exists in organisms with relevant data, such as Corydalis yanhusuo and Corydalis yanhusuo. - Dehydrocorydaline is a natural alkaloid derived from traditional Chinese medicines such as Corydalis yanhusuo [1,2,4]. - The antiproliferative effect of dehydrocorydaline on MCF-7 cells is mainly mediated by inducing mitochondrial-dependent apoptosis (manifested as changes in the Bax/Bcl-2 ratio and activation of caspase-3 and caspase-9) [1]. - The effect of dehydrocorydaline in promoting myogenic differentiation depends on the activation of the p38 MAPK signaling pathway [2]. - The analgesic effect of dehydrocorydaline: In an inflammatory pain model, naloxone (an opioid receptor antagonist) can reverse this pain, indicating that opioid receptors are involved in its analgesic mechanism [4].

C22H24NO4+

366.4303

366.171

30045-16-0

Dehydrocorydaline chloride;10605-03-5;Dehydrocorydaline (hydroxyl);Dehydrocorydaline nitrate;13005-09-9

34781

Solid

170-173℃

3.693

0

4

4

27

503

0

O(C([H])([H])[H])C1=C(C([H])=C([H])C2C1=C([H])[N+]1C([H])([H])C([H])([H])C3=C([H])C(=C(C([H])=C3C=1C=2C([H])([H])[H])OC([H])([H])[H])OC([H])([H])[H])OC([H])([H])[H]

RFKQJTRWODZPHF-UHFFFAOYSA-N

InChI=1S/C22H24NO4/c1-13-15-6-7-18(24-2)22(27-5)17(15)12-23-9-8-14-10-19(25-3)20(26-4)11-16(14)21(13)23/h6-7,10-12H,8-9H2,1-5H3/q+1

2,3,9,10-tetramethoxy-13-methyl-5,6-dihydroisoquinolino[2,1-b]isoquinolin-7-ium

Dehydrocorydaline; 13-Methylpalmatine

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: 50~73 mg/mL (136.5~199.2 mM)

Solubility in Formulation 1: ≥ 6.25 mg/mL (17.06 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 62.5 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.

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Solubility in Formulation 2: ≥ 6.25 mg/mL (17.06 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 62.5 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.

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 (Please use freshly prepared in vivo formulations for optimal results.)