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Method of dynamics analysis for compound in cell
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A compound and analysis method technology, applied in the field of in vitro analysis and evaluation, can solve problems such as huge development costs
Pending Publication Date: 2021-09-14
HITACHI HIGH-TECH CORP
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[0002] In the new drug development process, clinical trials ("human clinical trials") to verify the efficacy of drug administration in humans are required in the Pharmaceutical Affairs Act, and human clinical trials and animal experiments require huge development costs
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Embodiment 1
[0176] In this embodiment, according to figure 2 Steps 0 to 6 of the present invention will be described to apply the method for dynamic analysis of compounds of the present invention. Additionally, use image 3 Steps 4 to 6 will be described in detail.
[0177] It should be noted that the following examples are some examples for illustrating the present invention, and the specific conditions such as buffer solution, temperature, and time shown in this example can also use other conditions, as long as they have the same effect as a technical idea That's it.
[0178] In this example, the case where three test plots 1 to 3 are used will be described. In each test area, there are holding areas for holding cells, and these holding areas may use different containers for each test area, and each test area may be divided into one container having a plurality of holding areas.
[0179]
[0180] In this step, first, preparation and culture of cells for analyzing compound dynamic...
Embodiment 2
[0245] In Example 2, the case where a method different from that of Example 1 is used for determination and scoring of the distribution ratio in each component based on the measurement results will be described.
[0246] In this embodiment, by using image 3 The quantitative value of S# (corresponding to step 4) described, on the basis of the distribution ratios among the components shown in Figure 1 of Example 1, it is also possible to indicate whether the administered compound is easily retained in the cells or is difficult to retain. Information in cells can be evaluated more precisely. That is, the sum of steps 4, 5, and 6 ( image 3 , S#+B#+C#=(1)'+(2)'+(1)+(2)+(3)+(4)) as 100% of the administered compound amount. Think of this situation as "Graph 2". Therefore, 9 kinds of values (S1, S2, S3, B1, B2, B3, C1, C2, C3) obtained from the test plots of steps 4, 5, and 6 can be used as evaluation indexes.
[0247] Obtained from 9 values obtained by measuring the amount ...
Embodiment 3
[0256] In this example, the analysis was performed based on the amount of the intracellular compound transferred to the EA step (100%) (Figure 3). The amounts of compounds in test plots 1 to 3 should be the same in principle, but since experimental errors may occur, the analysis is performed based on the average value. The compound amount of each component shown in Figure 1 of Example 1 was applied to the following formula.
[0258] With this graph 3, more accurate analysis than graph 1 can be achieved.
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Abstract
The present invention provides a method and technique for using cells to analyze the dynamics of a compound such as a drug. More specifically, the present invention pertains to a method of dynamic analysis of a compound using cells, wherein the method comprises a CE step for adding a first buffer solution to a vessel that includes a subject compound and accommodates cells, adjusting the temperature inside the vessel to a first temperature and maintaining the temperature, and adjusting the temperature inside the vessel to a second temperature lower than the first temperature, the CE step involving measuring amounts S and S' of the compound in the first buffer solution at an interval t1, and calculating the rate of discharge of the compound from basal / basolateral surfaces of the cells on the basis of the amounts S and S' and the interval t1.
Description
technical field [0001] The present invention relates to invitro analysis and evaluation of the overall profile of compound dynamics including uptake, metabolism, and excretion of compounds. Background technique [0002] In the new drug development process, clinical trials (“human clinical trials”) to verify the efficacy of drug administration in humans are necessary in the pharmaceutical affairs law, and human clinical trials and animal experiments require huge development costs. Before transferring to such animal experiments and human clinical trials, it is preferable to verify the efficacy and toxicity of the candidate drug, and especially to evaluate it using an in vitro test system using human-derived cells. [0003] In order for a drug to be effective in the body, it is necessary that the drug temporarily taken up by the liver is converted into a metabolite, or it is directly excreted to the blood vessel side as the original compound (parent compound) without being meta...
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