Goldenseal (Hydrastis canadensis) 43525850
Poor permeability and absorption affect the activity of four alkaloids from Coptis CSV JSON

Coptidis rhizoma (Coptis) and its alkaloids exert various pharmacological functions in cells and tissues; however, the oral absorption of these alkaloids requires further elucidation. The present study aimed to examine the mechanism underlying the poor absorption of alkaloids, including berberine (BER), coptisine (COP), palmatine (PAL) and jatrorrhizine (JAT). An ultra-performance liquid chromatography (UPLC) method was validated for the determination of BER, COP, PAL and JAT in the above experimental medium. In addition, the apparent oil‐water partition coefficient (Po/w); apparent permeability coefficient (Papp), determined using a parallel artificial membrane permeability assay (PAMPA)plate; membrane retention coefficient (R %); and effect of P-glycoprotein (P-gp) inhibitor on the Papp of the four alkaloids were investigated. The intestinal absorption rate constant (Ka) and absorption percentage (A %) of the four alkaloids were also determined. The results of the present study demonstrated that the Po/w of the four alkaloids in 0.1 mol·l-1 HCl medium was significantly higher (P<0.01), compared with those of the alkaloids in phosphate buffer (pH 7.4). The Papp of BER was 1.0-1.2x10-6 cm·s-1, determined using a PAMPA plate, and the Papp of BER, COP, PAL and JAT decreased sequentially. The concentrations of the four alkaloids on the apical-to-basolateral (AP-BL) surface and the basolateral-to-apical (BL-AP) surface increased in a linear manner, with increasing concentrations between 10 and 100 μmol. In addition, the transportation ofBER on the BL‐AP surface was significantly faster (P<0.01), compared with that on the AP-BL surface and, following the addition of verpamil (a P-gp inhibitor), the Papp (AP-BL) of the four alkaloids increased, whereas the Papp (BL-AP) wassignificantly decreased (P<0.01). The rat intestinal perfusion experiment demonstrated that the four alkaloids were poorly absorbed; however, the Ka of BER was significantly higher, compared with the three other alkaloids. Furthermore, the A % and Ka provided evidence that the absorption of BER was increased in the jejunum, compared with in the ileum. In conclusion, the four alkaloids from Coptis appeared to be poorly absorbed, determined using a shake flask, pre‐coated PAMPA plates, a Caco-2 cell monolayer model and intestinal perfusion; however, absorption was higher in the jejunum than in the ileum. Among the four alkaloids, the permeability of BER was markedly higher than the others, and P‐gp efflux had a significant effect on the absorption of those alkaloids.

PMID: 26352530

26352530

1 . P-gp Permeability of Berberine Induced by Verapamil (id=NPDI-oYawZw)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used — not specified

berberine 19012197

verapamil

  • P-gp (ABCB1)

Cell system Caco-2 cells

Results

AP-BL/BL-AP

Berberine 7.30±0.02 

BER with verapamil 2.00±0.20 (p<0.01)
Table V

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

1 x 10^6 cells/mL

12 well

Transwell

21

Assay Conditions

30 min

37 deg C

7.4

30 μmol x l^(-1)

100 μmol x l^(-1)

Sigma-Aldrich (St. Louis, MO, USA)

1 . Efflux of Berberine through P-gp (id=NPDI-2ugVjg)

In Vitro Transporter Kinetics Experiment

No Transport Activity was detected.  Cutoff used —

Papp >1.5x10-6 cm·s-1 is considered to be highly penetrative; therefore, the Papp of the four alkaloids assessed in the present

berberine 19012197

  • P-gp (ABCB1)

Cell system Caco-2 cells

Results

Above results for 10 μmol

Papp A-B
20 μmol: 3.53
30 μmol: 2.85
60 μmol: 2.98
100 μmol: 2.68

Papp B-A
20 μmol: 31.30
30 μmol: 20.81
60 μmol: 25.30
100 μmol: 25.90

Ratio PappB-A / PappA-B
20 μmol: 8.87
30 μmol: 7.30
60 μmol: 8.49
100 μmol: 9.66

Table IV

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Cell Culture Conditions

5x10^4/cm^2

12-well plate

Transwell

21

Assay Conditions

60 min

37 degrees C

7.4

10, 20, 30, 60, 100 μmol*mL^(-1)