Componential Analyzer, Drug Efficacy Analyzer, and Analysis Method

Patent metadata

Field	Value
Publication number	US-2016209403-A1
Application number	US-201414914278-A
Country	US
Kind code	A1
Filing date	Nov 26, 2014
Priority date	Nov 29, 2013
Publication date	Jul 21, 2016
Grant date	—

Abstract

Application of the present invention enables quantification of fractions of candidate pharmaceutical compounds (a parent compound and its metabolites), one excreted to the basolateral (Basal/Basolateral)-side via transporters and by diffusion, one excreted to the lumen (Apical)-side, and one remained in the cells. This enables determination of the total amount of the administered candidate pharmaceutical compounds and the distribution ratio of the fractions. The kinetics of the administered candidate pharmaceutical compounds can be evaluated, thereby enabling in vitro screening of an enormous number of candidate pharmaceutical compounds for drug candidates exhibiting the efficacy. The object of the present invention is to provide an apparatus and method for understanding a total picture of pharmacokinetics in vitro by quantifying a fraction of basolateral (Basal/Basolateral) efflux, a fraction of lumen (Apical)-side excretion, and a fraction remaining in a cell of a drug which has been administered to the cell to determine the distribution ratio of each fraction.

First claim

1 . A componential analyzer, comprising:
a holding unit for holding a plurality of containers holding a predetermined cell; a temperature controlling unit for controlling temperatures in the plurality of containers; and an analyzing unit for measuring a component in the plurality of containers and analyzing the measured component, wherein the plurality of containers comprises at least a first container and a second container; the first container and the second container each contain a first buffer solution; the temperature controlling unit controls temperature in the first container and temperature in the second container so that the temperatures are different from each other; and the analyzing unit measures: an amount of the component excreted from a cell in the first container to the first buffer solution in the first container; and an amount of the component excreted from a cell in the second container to the first buffer solution in the second container; and the analyzing unit analyzes an amount of the component excreted via transporters in the cells.

2 . The componential analyzer according to claim 1 , wherein
the predetermined cell is a hepatocyte; the plurality of containers comprises at least the first container, the second container, and a third container; the third container contains a second buffer solution supplemented with a predetermined substance; the temperature controlling unit controls a first temperature in the first container and in the third container and a second temperature in the second container so that the second temperature is lower than the first temperature; the analyzing unit measures: an amount of the component excreted from the hepatocyte in the first container to the first buffer solution in the first container, an amount of the component excreted from the hepatocyte in the second container to the first buffer solution in the second container, and an amount of the component excreted from the hepatocyte in the third container to the second buffer solution in the third container; and the analyzing unit analyzes: an amount of the component excreted via transporters in the hepatocyte, an amount of the component excreted via bile canaliculi of the hepatocyte, and an amount of the component excreted via a route other than the transporters and the bile canaliculi of the hepatocyte.

3 . The componential analyzer according to claim 2 , further comprising a liquid-feeding unit for supplying or collecting a liquid in the plurality of containers,
wherein the liquid-feeding unit: supplies the plurality of containers with a component solution comprising the component, collects the component solution and then supplies the plurality of containers with a third buffer solution, and collects the third buffer solution and then supplies the first container and the second container with the first buffer solution and the third container with the second buffer solution; and the analyzing unit measures an amount of the component excreted into the third buffer solution from the hepatocyte contained in either of the plurality of containers.

4 . The componential analyzer according to claim 3 , wherein the third buffer solution and the first buffer solution are the same buffer solution.

5 . The componential analyzer according to claim 3 , wherein the temperature controlling unit controls temperature in at least one of the plurality of containers to be lower than the first temperature when the at least one of the plurality of containers contains the third buffer solution.

6 . The componential analyzer according to claim 1 , wherein the analyzing unit measures:
an amount of the component remaining in the hepatocyte in the third container and an amount of the component remaining in a hepatocyte in at least one container of the first container and the second container, and the analyzing unit analyzes an amount of the component remaining other than in the bile canaliculi of the hepatocyte.

7 . A drug component analyzer, comprising:
a holding unit for holding a plurality of containers each containing a hepatocyte having absorbed a drug; a liquid-feeding unit for supplying a liquid in the plurality of containers, a temperature controlling unit for controlling temperature in the plurality of containers; and an analyzing unit for measuring an amount of the drug in the plurality of containers and analyzing the measured drug, wherein the plurality of containers comprises a first container, a second container, and a third container; the liquid-feeding unit supplies the first container and the second container with a first buffer solution and the third container with a second buffer solution promoting excretion of the drug from bile canaliculus of the hepatocyte; the temperature controlling unit controls temperature in the second container to be lower than temperature in the first container and in the third container; the analyzing unit analyzes each of: an amount of the drug excreted into the first buffer solution from the hepatocyte in the first container, an amount of the drug excreted into the first buffer solution from the hepatocyte in the second container, an amount of the drug excreted into the second buffer solution from the hepatocyte in the third container, an amount of the drug excreted via transporters in the hepatocyte, an amount of the drug excreted via bile canaliculi of the hepatocyte, and an amount of the component excreted via a route other than the transporters and the bile canaliculi of the hepatocyte.

8 . A drug component analyzer, comprising:
a holding unit for holding a plurality of containers containing a hepatocyte having absorbed a drug; a liquid-feeding unit for supplying and discharging a liquid in the plurality of containers, a temperature controlling unit for controlling temperature in the plurality of containers; and an analyzing unit for measuring an amount of the drug in the plurality of containers and analyzing the measured drug, wherein the liquid-feeding unit: supplies the plurality of containers with the drug, discharges the drug from the plurality of containers and then supplies a pretreatment buffer solution, collects the pretreatment buffer solution from the plurality of containers and then supplies the first container and the second container of the plurality of culture containers with a first buffer solution, and supplies the third container with a second buffer solution promoting excretion of the drug from a bile canaliculus of the hepatocyte; the temperature controlling unit controls a first temperature in the first container and in the third container and a second temperature in the second container so that the second temperature is lower than the first temperature; the analyzing unit analyzes each of: an amount of the drug excreted into the pretreatment buffer solution from the hepatocytes in the plurality of containers, an amount of the drug excreted into the first buffer solution from the hepatocyte in the first container, an amount of the drug excreted into the first buffer solution from the hepatocyte in the second container, an amount of the drug excreted into the second buffer solution from the hepatocyte in the third container, an amount of the drug excreted from the hepatocyte during pretreatment of the hepatocyte, an amount of the drug excreted via transporters in the hepatocyte, an amount of the drug excreted via bile canaliculi of the hepatocyte, and an amount of the drug excreted via a route other than the transporters and the bile canaliculi of the hepatocyte.

9 . The drug component analyzer according to claim 8 , wherein the pretreatment buffer solution and the first buffer solution are th

Assignees

• Hitachi High Tech Corp

Inventors

• Takahashi Ryosuke
• Kanda Katsuhiro
• Shimizu Yusuke
• Ichikawa Hisashi

Classifications

• G01N33/15

  Medicinal preparations {; Physical properties thereof, e.g. dissolubility} · CPC title

• C12M41/46

  of cellular or enzymatic activity or functionality, e.g. cell viability · CPC title

• C12M25/00

  Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings · CPC title

• B01L3/5085

  for multiple samples, e.g. microtitration plates · CPC title

• G01N33/5067 Primary

  Liver cells · CPC title