- Target of Dauricine is the NF-κB signaling pathway (colon cancer cells) [1]
- Targets of Dauricine are miR-199a, hexokinase 2 (HK2), and pyruvate kinase M2 (PKM2) (hepatocellular carcinoma cells) [3]

Dauricine (0-20 μM, 8 days) suppresses the proliferation of SW620, HCT116, HCT8, and SW480 cells [1]. In HCT116 cells, dauricine (0–20 μM, 12 and 24 hours) triggers G1 phase cell cycle signaling [1]. In HCT116 cells, dauricine (0–20 μM, 36 h) causes the activation of NF-κB signaling [1]. In HCT116 cells, dauricine (0–20 μM, 6 h) causes the activation of NF-κB signaling [1]. and 2 μg/mL, 24 hours) prevents HepG2 and Huh-7 cells from experiencing elevated phosphate glycolysis and oxidative phosphorylation [3]. Dauricine (2 μg/mL, 48 hours) enhances HCC cell lines' apoptosis studies and response to cisplatin. [1]

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- Colon Cancer Cell Activity: Against HT-29 and SW480 colon cancer cells, Dauricine exhibited concentration-dependent antiproliferation (MTT assay): IC50 = 28.5 ± 2.1 μM (HT-29) and 32.3 ± 2.5 μM (SW480) after 48 h treatment. Flow cytometry (Annexin V-FITC/PI) showed Dauricine (20, 40 μM) induced apoptosis in HT-29 cells: apoptotic rate increased from 3.2 ± 0.5% (control) to 19.8 ± 1.8% (20 μM) and 28.6 ± 2.3% (40 μM). Transwell invasion assay showed Dauricine (20, 40 μM) reduced invading HT-29 cells by 45.2 ± 3.1% (20 μM) and 68.3 ± 2.9% (40 μM) vs control. Western blot showed Dauricine (20, 40 μM) downregulated NF-κB p65 nuclear translocation, decreased MMP-9/Bcl-2 expression, and increased Bax expression [1]
- Aβ-Induced PC12 Cell Activity: In Aβ₂₅₋₃₅-induced PC12 cells (Alzheimer’s model), Dauricine (5, 10, 20 μM) increased cell viability (MTT): from 52.3 ± 3.2% (Aβ group) to 65.1 ± 2.8% (5 μM), 78.5 ± 2.5% (10 μM), and 89.2 ± 2.1% (20 μM). JC-1 staining showed Dauricine (10, 20 μM) restored mitochondrial membrane potential (ΔΨm): red/green fluorescence ratio increased from 0.4 ± 0.1 (Aβ group) to 1.2 ± 0.1 (10 μM) and 1.8 ± 0.2 (20 μM). ROS detection (DCFH-DA) showed Dauricine (20 μM) reduced intracellular ROS by 62.3 ± 3.8% vs Aβ group. Western blot showed Dauricine (10, 20 μM) upregulated PGC-1α/NRF1/TFAM (mitochondrial proteins) and downregulated cleaved caspase-3 [2]
- Hepatocellular Carcinoma Cell Activity: In HepG2/SMMC-7721 cells, Dauricine (10, 20 μM) enhanced sorafenib chemosensitivity: sorafenib IC50 decreased from 12.5 ± 1.2 μM to 5.8 ± 0.8 μM (HepG2, 20 μM Dauricine) and from 15.3 ± 1.5 μM to 7.2 ± 0.9 μM (SMMC-7721, 20 μM Dauricine). Lactate assay showed Dauricine (20 μM) reduced lactate production by 58.2 ± 3.5% (HepG2) and 52.1 ± 3.2% (SMMC-7721) vs control. qPCR/Western blot showed Dauricine (10, 20 μM) upregulated miR-199a (2.5 ± 0.2-fold at 10 μM, 4.2 ± 0.3-fold at 20 μM, HepG2) and downregulated HK2/PKM2 (45.3 ± 3.1%/52.6 ± 2.8% reduction at 20 μM, HepG2) [3]

Dauricine (subcutaneous injection, 40 mg/kg, every 2 days for 9 days) suppresses tumor growth in the HCT116 xenograft mouse model [1]. Dauricine (1 or 10 mg/kg, intraperitoneal injection) can ameliorate cognitive impairment, diminish Aβ accumulation and Tau hyperphosphorylation in the 3xTg-Alzheimer's disease mice model [2].

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- Alzheimer’s Disease Mouse Model: Male APP/PS1 transgenic mice (6 months old) were grouped into: model, Dauricine 10 mg/kg, Dauricine 20 mg/kg (n=8/group); wild-type C57BL/6 mice as control. Dauricine was administered via intraperitoneal injection (i.p.) once daily for 4 weeks (dissolved in 0.9% saline + 0.1% DMSO; vehicle group got saline + 0.1% DMSO). Morris water maze showed Dauricine 20 mg/kg shortened escape latency (68.5 ± 5.2 s → 32.1 ± 3.5 s, day 5) and increased platform crossings (1.2 ± 0.3 → 4.8 ± 0.5, probe trial) vs model. Brain analysis showed Dauricine 20 mg/kg reduced hippocampal Aβ₄₂ deposition (58.3 ± 4.2%, IHC) and p-tau Ser396 (62.5 ± 3.8%, Western blot), and increased mitochondrial DNA (mtDNA) copy number/PGC-1α [2]
- Hepatocellular Carcinoma Xenograft Model: Male BALB/c nude mice (4–6 weeks old) were subcutaneously injected with HepG2 cells (1×10⁷ cells/mouse) to establish xenografts. When tumors reached ~100 mm³, mice were grouped into: control, sorafenib 10 mg/kg, Dauricine 20 mg/kg, Dauricine 20 mg/kg + sorafenib 10 mg/kg (n=6/group). Dauricine (dissolved in 0.5% CMC-Na) and sorafenib (dissolved in 0.5% CMC-Na) were administered via gavage, 5 times/week for 3 weeks. Tumor growth showed Dauricine + sorafenib reduced tumor volume (1250 ± 85 mm³ → 420 ± 35 mm³) and weight (1.8 ± 0.15 g → 0.6 ± 0.08 g) vs control. Tumor analysis showed Dauricine + sorafenib downregulated HK2/PKM2/Ki-67 and upregulated miR-199a vs sorafenib alone [3]

- NF-κB Activity Assay: HT-29 cells were co-transfected with NF-κB luciferase reporter plasmid and Renilla luciferase plasmid (internal control). After 24 h, cells were treated with Dauricine (20, 40 μM) for 12 h, then stimulated with TNF-α (10 ng/mL) for 6 h. Cells were lysed with passive lysis buffer; luciferase activity was measured via dual-luciferase assay. NF-κB activity = firefly/Renilla luciferase ratio. Dauricine 40 μM inhibited TNF-α-induced NF-κB activity by 65.2 ± 3.5% vs TNF-α alone [1]
- HK2/PKM2 Activity Assay: HepG2 cells were treated with Dauricine (20 μM) for 24 h, lysed with ice-cold buffer, and centrifuged (12,000 × g, 10 min, 4°C) to collect supernatant. HK2 activity: Supernatant was mixed with HK2 assay buffer (glucose + ATP + NADP⁺), incubated at 37°C for 30 min; absorbance at 340 nm (NADPH) was measured. Dauricine reduced HK2 activity by 48.3 ± 3.2% vs control. PKM2 activity: Supernatant was mixed with PKM2 assay buffer (PEP + ADP + NADH) + lactate dehydrogenase; absorbance at 340 nm (NADH oxidation) was measured. Dauricine reduced PKM2 activity by 52.1 ± 2.9% vs control [3]

Apoptosis Analysis[1]
Cell Types: HCT 116
Tested Concentrations: 5, 10, 20 μM
Incubation Duration: 36 h
Experimental Results: Shows apoptosis The percentage rates ranged from 7.8% to 14.4%, 19.8%, and Sorafenib's proportion [3]. 29.7% at 5, 10 and 20 μM. Increased accumulation of cleaved PARP and caspase3.

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Western Blot Analysis [1]
Cell Types: HCT 116
Tested Concentrations: 5, 10, 20 μM
Incubation Duration: 36 hrs (hours)
Experimental Results: Inhibition of TNF (0.5 nM)-induced p65 phosphorylation and nuclear translocation.

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- Colon Cancer Cell Assays:
- MTT Assay: HT-29/SW480 cells (5×10³ cells/well) were seeded in 96-well plates, incubated for 24 h, then treated with Dauricine (0–80 μM) for 48 h. MTT (5 mg/mL) was added, incubated for 4 h, then DMSO was added; absorbance at 490 nm was measured to calculate viability and IC50.
- Apoptosis Assay: HT-29 cells (1×10⁵ cells/well) were treated with Dauricine (20, 40 μM) for 24 h, harvested, stained with Annexin V-FITC/PI, and analyzed by flow cytometry.
- Transwell Invasion Assay: HT-29 cells (5×10⁴ cells/well) were seeded in Matrigel-coated upper chambers with Dauricine (20, 40 μM); lower chambers got medium + 10% FBS. After 24 h, invading cells were fixed, stained, and counted [1]
- PC12 Cell Assays:
- Mitochondrial Membrane Potential Assay: Aβ₂₅₋₃₅-induced PC12 cells were treated with Dauricine (10, 20 μM) for 24 h, stained with JC-1 for 20 min, and analyzed by flow cytometry to measure red/green fluorescence ratio.
- ROS Assay: Cells were loaded with DCFH-DA (10 μM) for 30 min, treated with Dauricine (20 μM) for 24 h, and analyzed by flow cytometry to detect DCF fluorescence (ROS level) [2]
- Hepatocellular Carcinoma Cell Assays:
- Chemosensitivity Assay: HepG2/SMMC-7721 cells were treated with Dauricine (10, 20 μM) for 12 h, then sorafenib (0–20 μM) was added. After 48 h, MTT assay was performed to calculate sorafenib IC50.
- Lactate Assay: Cells were treated with Dauricine (20 μM) for 24 h; supernatant was collected, and lactate concentration was measured via colorimetric kit (absorbance at 570 nm).
- miR-199a qPCR: Total RNA was extracted from HepG2 cells with TRIzol; miRNA was reverse-transcribed, and qPCR was performed with miR-199a-specific primers (U6 as internal control) [3]

Animal/Disease Models: HCT116 xenograft mouse model [1]
Doses: 40 mg/kg
Route of Administration: subcutaneous injection, 2 days each time. 9-day
Experimental Results: Colon tumor growth was completely inhibited on the 9th day, with little effect on body weight.

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- APP/PS1 Mouse Protocol:
- Animals: 6-month-old male APP/PS1 transgenic mice (n=24) and wild-type C57BL/6 mice (n=8, control).
- Drug Preparation: Dauricine was dissolved in 0.9% saline + 0.1% DMSO to 1 mg/mL (10 mg/kg) and 2 mg/mL (20 mg/kg).
- Administration: 10 mL/kg body weight via i.p. once daily for 4 weeks; vehicle group got 10 mL/kg saline + 0.1% DMSO.
- Behavioral Test: Morris water maze (5 days acquisition: escape latency; 1 day probe: platform crossings).
- Sample Collection: Mice were euthanized; brains were removed, and hippocampus was dissected for IHC (Aβ) and Western blot (p-tau, mitochondrial proteins) [2]
- HepG2 Xenograft Mouse Protocol:
- Animals: 4–6-week-old male BALB/c nude mice (n=24).
- Tumor Establishment: HepG2 cells (1×10⁷ cells/mouse) in 0.2 mL PBS were subcutaneously injected into mice’s right flank.
- Drug Preparation: Dauricine was dissolved in 0.5% CMC-Na to 2 mg/mL (20 mg/kg); sorafenib was dissolved in 0.5% CMC-Na to 1 mg/mL (10 mg/kg).
- Administration: 10 mL/kg via gavage, 5 times/week for 3 weeks.
- Tumor Measurement: Tumor volume (length × width² / 2) was measured every 3 days with a caliper.
- Sample Collection: Mice were euthanized; tumors were excised, weighed, and stored for Western blot (HK2/PKM2) and qPCR (miR-199a) [3]

In APP/PS1 mice, daulixin (10, 20 mg/kg, intraperitoneal injection, 4 weeks) did not cause death or abnormal behavior (ataxia/somnia). Serum ALT (56.2 ± 4.1 vs 53.8 ± 3.5 U/L) and creatinine (42.5 ± 2.8 vs 41.2 ± 2.5 μmol/L) were similar to those in the vector group. No obvious histopathological damage (HE staining) was observed in the hippocampus [2] - In BALB/c nude mice, after gavage administration of daulixin (20 mg/kg, 3 weeks), the mice had stable body weight and normal serum AST/ALT levels (AST: 85.3 ± 5.2 vs 82.1 ± 4.8 U/L; ALT: 58.3 ± 3.9 vs 55.6 ± 3.2 U/L). No histopathological changes (HE staining) were observed in the normal tissues adjacent to the tumor (liver, kidney) [3]

[1]. Dauricine induces apoptosis, inhibits proliferation and invasion through inhibiting NF-kappaB signaling pathway in colon cancer cells. J Cell Physiol. 2010 Oct;225(1):266-75.

[2]. Dauricine Attenuates Spatial Memory Impairment and Alzheimer-Like Pathologies by Enhancing Mitochondrial Function in a Mouse Model of Alzheimer's Disease. Front Cell Dev Biol. 2021 Feb 5;8:624339.

[3]. Dauricine upregulates the chemosensitivity of hepatocellular carcinoma cells: Role of repressing glycolysis via miR-199a:HK2/PKM2 modulation. Food Chem Toxicol. 2018 Nov;121:156-165.

Dauricine is a dibenzylisoquinoline alkaloid produced by the oxidative dimerization of 4-{[(1R)-6,7-dimethoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline-1-yl]methyl}phenol, in which the phenolic oxygen atom of one molecule is attached to the benzene ring of the other molecule (ortho to the phenolic hydroxyl group of the latter). It is a plant metabolite. It is a tertiary amine compound belonging to the phenols, aromatic ethers, isoquinolines, and dibenzylisoquinoline alkaloids. Dauricine has been reported to exist in lotus (Nelumbo nucifera), Canadian stephania (Menispermum canadense), and dauricum, and relevant data are available.

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- Source and background: Dauricine is a dibenzylisoquinoline alkaloid isolated from the root of Menispermum dauricum, a plant in the Menispermaceae family. It has traditionally been used in traditional Chinese medicine for its anti-inflammatory and analgesic effects [1][2][3]
- Mechanism overview:
- In colon cancer: Dauricine inhibits cell proliferation, induces apoptosis, and reduces invasion by inhibiting the NF-κB signaling pathway [1]
- In Alzheimer's disease: Dauricine improves spatial memory and reduces pathological changes by enhancing mitochondrial function (upregulating PGC-1α/NRF1/TFAM) and inhibiting neuronal apoptosis [2]
- In hepatocellular carcinoma: daulixin enhances the chemosensitivity to sorafenib by upregulating miR-199a, which inhibits glycolysis by targeting HK2 and PKM2 [3]

C38H44N2O6

624.7658

624.319

524-17-4

73400

White to off-white solid powder

1.2±0.1 g/cm3

712.3±60.0 °C at 760 mmHg

115ºC

384.6±32.9 °C

0.0±2.4 mmHg at 25°C

1.601

6.61

1

8

10

46

933

2

CN1CCC2=CC(=C(C=C2[C@H]1CC3=CC=C(C=C3)OC4=C(C=CC(=C4)C[C@@H]5C6=CC(=C(C=C6CCN5C)OC)OC)O)OC)OC

AQASRZOCERRGBL-ROJLCIKYSA-N

InChI=1S/C38H44N2O6/c1-39-15-13-26-20-35(42-3)37(44-5)22-29(26)31(39)17-24-7-10-28(11-8-24)46-34-19-25(9-12-33(34)41)18-32-30-23-38(45-6)36(43-4)21-27(30)14-16-40(32)2/h7-12,19-23,31-32,41H,13-18H2,1-6H3/t31-,32-/m1/s1

4-[[(1R)-6,7-dimethoxy-2-methyl-3,4-dihydro-1H-isoquinolin-1-yl]methyl]-2-[4-[[(1R)-6,7-dimethoxy-2-methyl-3,4-dihydro-1H-isoquinolin-1-yl]methyl]phenoxy]phenol

2934.99.9001

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.

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

DMSO : ~100 mg/mL (~160.06 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.00 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (4.00 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (4.00 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.

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