中文 English
In Vitro Study for Metabolizing Enzymes Inhibition

In Vitro Study for Metabolizing Enzymes Inhibition

全国热线
021-50800743

Metabolic enzyme inhibition tests were conducted to evaluate the inhibitory effects of the tested products on major subenzymes such as CYP450, UGT, SULT or CES, and to understand their direct inhibition or time-dependent inhibition (TDI) effects, so as to evaluate their possible metabolic interactions.

Incubation system

Cell component: liver cells

Subcellular components: microsomes, S9 or homogenates of tissues such as liver, intestine, lung or kidney

Gene recombination enzyme system

Species

Man

Tested enzymes

CYP450 enzyme: CYP1A2, 2B6, 2C8, 2C9, 2C19, 2D6 and 3A, etc

UGT enzymes: UGT1A1, 1A3, 1A4, 1A6, 1A9, 2B7 and 2B15, etc

SULT enzymes: SULT1A1, 1A2, 1A3, 1B1, 1E1 and 2A1, etc

CES enzymes: CES1 and CES2

Other: Other enzymes or subtypes such as AADAC, FMO are available upon request

Test concentrations

5 to 7 samples were designed according to known efficacy and toxicity data

Probe substrate concentration

1 or 1-5 (direct inhibition constant determination), based on the probe substrate Km value design

Incubation system Protein concentration

Subcellular component: 0.1-1mg /mL (different concentrations available on request)

Cell composition: 0.1-2×106 cells/mL

Recombinant enzyme system: 250-500 pmol/mL or 0.1-1 mg/mL

Incubation time

Direct inhibition or TDI IC50 shift: One pre-incubation time point, one incubation time point

TDI KI and Kinact measurements: 2 to 5 time points for pre-incubation, one time point for formal incubation

Available on request

Coenzyme factor

NADPH or UDPGA (Other Coenzyme factors are available upon request)

Analytical method

LC-MS/MS

Data presentation

Relative production of probe substrate metabolites

Relative activity (% of NC)

Median inhibitory concentration (IC50)

Inhibitory activity (%)

IC50 migration

Inhibition constant (Ki)

Maximum deactivation rate Constant (Kinact)

Concentration of free inhibitors (KI) leading to 50% maximum inactivation


您的信息对我们十分重要,请填写完整,以便更好的为您服务,谢谢!
更多详情

  • 上一条
  • 下一条