Acetylcholinesterase (AChE) (IC₅₀ = 1.07 µM)[1]
Butyrylcholinesterase (BChE) (IC₅₀ = 6.03 µM)[1]
β-site amyloid precursor protein cleaving enzyme 1 (BACE1) (IC₅₀ = 8.55 µM, Ki = 10.0, non-competitive inhibition)[1]
Peroxynitrite (ONOO⁻) (IC₅₀ = 16.83 µM for scavenging activity)[1]

In 3T3-L1 adipocytes, epiberberine (0, 12.5, 25 or 50 μM) suppresses cellular triglyceride accumulation in a dose-dependent manner, with an IC50 of 52.8 μM [2]. During the initial stages of 3T3-L1 adipocyte development, epiberberine (12.5-50 μM) suppresses the Raf/MEK1/ERK1/2 and AMPKα/Akt pathways. The concentration-dependent inhibition of glucose absorption in HepG2 cells is caused by epiberberine (0.2, 1, 5 μg/mL) [3].

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Epiberberine exhibited potent inhibitory activity against AChE with an IC₅₀ value of 1.07 µM, and against BChE with an IC₅₀ of 6.03 µM.[1]
Epiberberine showed good, dose-dependent inhibitory activity against BACE1 with an IC₅₀ of 8.55 µM and a Ki value of 10.0, indicating non-competitive inhibition.[1]
Epiberberine demonstrated ONOO⁻ scavenging activity with an IC₅₀ of 16.83 µM.[1]
In a total reactive oxygen species (ROS) inhibitory assay using rat kidney post-mitochondrial fractions, Epiberberine did not show significant inhibitory activity within the tested concentrations (IC₅₀ > 100 µM).[1]

In KK-Ay mice, oral administration of epiberberine (225 mg/kg) for 40 days results in decreased body weight, food consumption, water intake, and urine output [3].

The BACE1 inhibitory activity was assessed using a fluorescence resonance energy transfer (FRET) assay kit. Briefly, a reaction mixture containing assay buffer, BACE1 enzyme, a fluorogenic substrate peptide, and the test sample dissolved in DMSO was incubated for 60 minutes at 25°C in the dark. The increase in fluorescence resulting from substrate cleavage was measured using a microplate spectrofluorometer. The percentage inhibition was calculated by comparing the fluorescence change in sample wells to that in control wells. The concentration causing 50% inhibition (IC₅₀) was determined from a log-dose inhibition curve. For kinetic analysis to determine the inhibition mechanism (Ki and mode), the assay was performed at three different concentrations of Epiberberine (2, 4, 10 µM) against varying substrate concentrations (150, 250, 375 nM). The Ki value was determined from the Dixon plot.[1]
The inhibitory activities against AChE and BChE were measured using a spectrophotometric method. The reaction mixture contained sodium phosphate buffer, the test sample dissolved in DMSO, and the enzyme (AChE or BChE). After incubation, the reaction was initiated by adding the substrate (acetylthiocholine iodide for AChE or butyrylthiocholine chloride for BChE) and 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB). The hydrolysis of the substrate was monitored by following the formation of the yellow 5-thio-2-nitrobenzoate anion at 412 nm over 15 minutes. Percent inhibition was calculated, and the IC₅₀ values were determined from log-dose inhibition curves.[1]
ONOO⁻ scavenging activity was measured by monitoring the oxidation of dihydrorhodamine 123 (DHR 123). Test samples dissolved in DMSO were mixed with buffer and DHR 123. Authentic ONOO⁻ was added, and after 5 minutes, the fluorescence intensity of oxidized DHR 123 was measured using a microplate fluorescence reader. The percentage inhibition of DHR 123 oxidation was calculated.[1]
Total ROS inhibitory activity was assessed using the ROS-sensitive fluorescence indicator 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). Test samples dissolved in DMSO were added to a rat kidney post-mitochondrial fraction in buffer, followed by loading with DCFH-DA. The fluorescence of the oxidation product (DCF) was measured over 30 minutes using a microplate fluorescence spectrophotometer.[1]

[1]. Anti-Alzheimer and antioxidant activities of Coptidis Rhizoma alkaloids. Biol Pharm Bull. 2009 Aug;32(8):1433-8.

[2]. Anti-adipogenic effect of epiberberine is mediated by regulation of the Raf/MEK1/2/ERK1/2 and AMPKα/Akt pathways. Arch Pharm Res. 2015 Dec;38(12):2153-62.

[3]. Antihyperglycemia and Antihyperlipidemia Effect of Protoberberine Alkaloids From Rhizoma Coptidis in HepG2 Cell and Diabetic KK-Ay Mice. Drug Dev Res. 2016 Jun;77(4):163-70.

Epidermalonine is a protoberberine alkaloid isolated from Coptis chinensis. [1]
The methylenedioxy group on its D ring is considered a key factor in its BACE1 inhibitory activity. [1]
It has a variety of activities related to anti-Alzheimer's disease, including cholinesterase inhibition, BACE1 inhibition and antioxidant (ONOO⁻ scavenging) activities. [1]

C20H18CLNO4

371.814224720001

371.092

889665-86-5

Epiberberine;6873-09-2

71467079

Yellow to orange solid powder

0

5

2

26

488

0

[Cl-].O1COC2=CC=C3C=C4C5C=C(C(=CC=5CC[N+]4=CC3=C12)OC)OC

DGRBIBRPLDAHJH-UHFFFAOYSA-M

InChI=1S/C20H18NO4.ClH/c1-22-18-8-13-5-6-21-10-15-12(3-4-17-20(15)25-11-24-17)7-16(21)14(13)9-19(18)23-2;/h3-4,7-10H,5-6,11H2,1-2H3;1H/q+1;/p-1

8,9-Dimethoxy-11,12-dihydro-[1,3]dioxolo[4,5-h]isoquinolino[2,1-b]isoquinolin-13-ium chloride

2934.99.9001

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, avoid exposure to moisture.

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

DMSO : ~25 mg/mL (~67.24 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.72 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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 (Please use freshly prepared in vivo formulations for optimal results.)