CN109765348A - A kind of accurately external medicine heart peace comments method - Google Patents
A kind of accurately external medicine heart peace comments method Download PDFInfo
- Publication number
- CN109765348A CN109765348A CN201811478355.6A CN201811478355A CN109765348A CN 109765348 A CN109765348 A CN 109765348A CN 201811478355 A CN201811478355 A CN 201811478355A CN 109765348 A CN109765348 A CN 109765348A
- Authority
- CN
- China
- Prior art keywords
- frequency
- stimulation
- action potential
- perfusion
- drug
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention discloses a kind of accurately external medicine heart peace and comments method, main includes action potential duration and frequency correlation coefficient K value in relation to (slope that K value is Action Potential Duration and frequency matched curve), the invention experiments have shown that, k-factor is related to drug to action potential frequency dependence as a result, this result will have a direct impact on the cardiac safety risk evaluation result of drug.Therefore, the risk that can more precisely, more effectively assess drug arrhythmia cordis is experimentally standardized using this COEFFICIENT K, and the appearance of arrhythmia cordis false positive or false negative before clinical drug is prevented, higher efficiency, high-throughput drug screening may be implemented in k-factor standardization.
Description
Technical field
The present invention relates to drug safety assessment technology field, specifically a kind of accurately external medicine heart pacifies the side of commenting
Method.
Background technique
Cardiac safety assessment be each kind new medicine enter before clinical test one for need to be considered when it is important with animal or dynamic
Experimental arrangement based on object tissue.In the past few years, some drugs for having put into clinical application are because of its potential Amplatzer duct occluder
Property or can cause serious arrhythmia cordis and be forced approval is recalled or postponed from market to enter market, such as astemizole, Te Feina
Fixed, Cisapride qualifying pa sand magnitude.Human drugs registration technology require international coordination meeting (ICH) clear stipulaties: new drug into
Before entering clinical application, it is necessary to carry out the cardiac safety assessment of animal and human body.Currently, mainly being detected by hERG and QT interphase
Predict arrhythmia cordis risk, but its related mechanism is studies have shown that hERG retardance and proarrhythmia risk and clinic QTc extension
Correlation it is not quite identical, i.e., using hERG detection prediction QT interval prolongation reliability it is not high.However QT interval prolongation
Also not necessarily cause fatal arrhythmia, such as Alfuzosin, phenobarbital and ranolazine that can extend QT interphase, but not
It will lead to torsades de pointes (TdP).QTc is obtained by ECG, many studies have shown that ECG can not be complete
Site preparation reflect the complete repolarization information of heart, therefore using QT interphase as measurement heart entirety repolarization Testing index can
It is not high by property.However, research discovery QT extends heterogeneous related, the action potential duration to action potential duration
Heterogeneous is again to lead to the important foundation of arrhythmia cordis, and body surface ECG can not detect this heterogeneity, therefore, preclinical
The related experiment for increasing drug delay ventricular bipolar is very urgent.The related experiment of drug delay ventricular bipolar mainly passes through movement
Potential Examination detects, and up to now, the selection for restoring for 90% time for ventricular repolarisation without ready patterns to follow, in this way will lead to
For assessment medicine heart risk, there is a certain error for we.Since whether evaluation drug is one quite multiple safely to heart
Miscellaneous process, many times, since there are many not rigorous probability to increase false positive or false negative of experimental design, it can be with
It is abandoned drug with fine potentiality itself therefore, this will can all cause measure to any pharmacy corporation
Loss, therefore formulate a kind of accurately external medicine heart peace and comment method extremely urgent.
Summary of the invention
To solve the above-mentioned problems, the invention discloses a kind of accurately external medicine heart peaces to comment method.
The technical solution of the present invention is as follows: a kind of accurately external medicine heart peace comments method, including the following steps:
(1) action potential Frequency Design: stimulate Purkinje fiber in difference with three groups of different stimulated frequencies include 60 pulses
The action potential started under frequency of stimulation, three groups of frequency of stimulation periods are respectively 2000ms, 1000ms and 500ms；
(2) action potential records: rabbit Purkinje fiber being placed in perfusion bath, by the speed perfusion Tai Shi of 5 mL/min
Liquid, using neonatal canine method, Purkinje fiber first with 1 basis s stimulation perimeter perfusion Tai Shi solution 30 minutes with
On, so that it is stablized and restore normal excitability；The glass microelectrode insertion sample of the KCl solution of 3 M will be inoculated with after 30 minutes,
Action potential is drawn, 1 Hz of frequency of stimulation, 10 kHz of sample frequency, the overall process of every an example observation experiment project is same
It is completed on cell, Purkinje fiber perfusion solvent control 15 minutes after stabilization, action potential is steady under the frequency of stimulation of 1 Hz
It after fixed, then give the frequency stimulation of 2000 ms, 1000 ms and 500 ms and records 60 stable action potentials respectively, then change
It closes the concentration of object from low to high to be successively carried out continuously Purkinje fiber perfusion 15 minutes, repeats above-mentioned frequency of stimulation note respectively
Action potential is recorded, it is identical when stimulation programs are with solvent control.
Usefulness of the present invention: having determined the optimum range to Action Potential Duration selection, establishes dynamic outside standardization body
Object heart peace comments platform that can more precisely, more effectively assess drug to the security risk of heart, and drug is enabled to obtain high pass
Amount, efficient screening, to preferably prevent the appearance of the false negative or false positive of clinical drug arrhythmia cordis.
Detailed description of the invention
Fig. 1 is action potential duration and k-factor relational graph in the present invention；
Fig. 2 is the selection figure of action potential duration optimum range in the present invention；
Fig. 3 be in the present invention k-factor to dofetillide action effect influence diagram；
Fig. 4 be in the present invention k-factor to the influence diagram of dofetillide frequency dependence；
Fig. 5 is the related coefficient K of drug in the present invention (APD90) and the relational graph of its concentration；
Fig. 6 is the related coefficient K of drug in the present invention (APD60) and the relational graph of its concentration.
Specific embodiment
In order to deepen the understanding of the present invention, it describes the specific embodiments of the present invention in detail with reference to the accompanying drawing, the reality
It applies example for explaining only the invention, does not restrict the protection scope of the present invention.
A kind of accurately external medicine heart peace comments method, including the following steps:
(1) action potential Frequency Design: stimulate Purkinje fiber in difference with three groups of different stimulated frequencies include 60 pulses
The action potential started under frequency of stimulation, three groups of frequency of stimulation periods are respectively 2000ms, 1000ms and 500ms；
(2) action potential records: rabbit Purkinje fiber being placed in perfusion bath, by the speed perfusion Tai Shi of 5 mL/min
Liquid, using neonatal canine method, Purkinje fiber first with 1 basis s stimulation perimeter perfusion Tai Shi solution 30 minutes with
On, so that it is stablized and restore normal excitability；Present invention discover that the Pu Ken of only 3 180 ms of Action Potential Duration APD90
Wild Fibers Action Potential can be only used for subsequent experiment (ensure the accuracy tested).The KCl of 3 M will be inoculated with after 30 minutes
The glass microelectrode of solution is inserted into sample, draws action potential, 1 Hz of frequency of stimulation, 10 kHz of sample frequency, and every an example is seen
The overall process for examining experimental project is completed on same cell, Purkinje fiber perfusion solvent control 15 minutes after stabilization, 1
After action potential is stablized under the frequency of stimulation of Hz, then gives the frequency stimulation of 2000 ms, 1000 ms and 500 ms and record respectively
60 stable action potentials, then the concentration of compound from low to high is successively carried out continuously perfusion 15 to Purkinje fiber and divides
Clock repeats above-mentioned frequency of stimulation operation of recording current potential respectively, identical when stimulation programs are with solvent control.
Action potential record before clinical drug in heart methods of risk assessment has mainly been carried out an experiment by the present invention
Standardized to explore and establish standardized assay method according to result, experimental result is as follows:
(1) as shown in Figure 1, X-axis indicates frequency of stimulation (2000ms, 1000ms and 600ms), Y-axis indicates the ventricular repolarisation time,
The difference for analyzing APD90>180ms and APD90<180ms coefficient of correspondence K, as a result, it has been found that when APD90>180ms, K value is
28.1, then show good frequency dependence；When APD90 < 180ms, K value is 3.6, then frequency dependence is not present；I.e.
APD90 value is bigger, and COEFFICIENT K is then bigger, otherwise smaller.
(2) as shown in Fig. 2, APD90 under X-axis 1Hz, Y-axis indicates the related coefficient (K) of frequency, by all control
Analysis of summarizing is tested, as a result, it has been found that K shows ascendant trend when APD90 > 180ms.
(3) as shown in figure 3, APD90 under X-axis 1Hz, Y-axis indicate the percentage variation that 1 μM of dofetillide acts on it
Value, as a result, it has been found that dofetillide also enhances the effect of action potential therewith with the increase of ventricular repolarisation time.
(4) as shown in figure 4, X-axis indicates frequency of stimulation (2000ms, 1000ms and 600ms), Y-axis indicates ventricular repolarisation
Time by the APD90 of selection at 1 hz, respectively>180ms and<180ms, then is given although corresponding dofetillide acts on
The comparison of frequency correlation coefficient K is carried out, as a result, it has been found that obviously to obtain K value than APD90<180ms big for the K value of APD90>180ms.
Claims (1)
1. a kind of accurately external medicine heart peace comments method, characterized in that it comprises the following steps:
(1) action potential Frequency Design: stimulate Purkinje fiber in difference with three groups of different stimulated frequencies include 60 pulses
The action potential started under frequency of stimulation, three groups of frequency of stimulation periods are respectively 2000ms, 1000ms and 500ms；
(2) action potential records: rabbit Purkinje fiber being placed in perfusion bath, by the speed perfusion Tai Shi of 5 mL/min
Liquid, using neonatal canine method, Purkinje fiber first with 1 basis s stimulation perimeter perfusion Tai Shi solution 30 minutes with
On, so that it is stablized and restore normal excitability；The glass microelectrode insertion sample of the KCl solution of 3 M will be inoculated with after 30 minutes,
Action potential is drawn, 1 Hz of frequency of stimulation, 10 kHz of sample frequency, the overall process of every an example observation experiment project is same
It is completed on cell, Purkinje fiber perfusion solvent control 15 minutes after stabilization, action potential is steady under the frequency of stimulation of 1 Hz
It after fixed, then give the frequency stimulation of 2000 ms, 1000 ms and 500 ms and records 60 stable action potentials respectively, then change
It closes the concentration of object from low to high to be successively carried out continuously Purkinje fiber perfusion 15 minutes, repeats above-mentioned frequency of stimulation note respectively
Action potential is recorded, it is identical when stimulation programs are with solvent control.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811478355.6A CN109765348A (en) | 2018-12-05 | 2018-12-05 | A kind of accurately external medicine heart peace comments method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811478355.6A CN109765348A (en) | 2018-12-05 | 2018-12-05 | A kind of accurately external medicine heart peace comments method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109765348A true CN109765348A (en) | 2019-05-17 |
Family
ID=66450238
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811478355.6A Pending CN109765348A (en) | 2018-12-05 | 2018-12-05 | A kind of accurately external medicine heart peace comments method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109765348A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109856349A (en) * | 2018-12-05 | 2019-06-07 | 南通科瑞斯生物医药科技有限公司 | A kind of optimization method of high throughput hERG assessment cardiac safety risk |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU1790768C (en) * | 1989-09-13 | 1993-01-23 | Л.В.Бобровников | Method of nerve cell membrane permeability determination |
| CN102883653A (en) * | 2010-03-22 | 2013-01-16 | 莱斯特大学 | Method and apparatus for evaluating cardiac function |
| CN104237308A (en) * | 2013-06-09 | 2014-12-24 | 国家纳米科学中心 | In-vitro medicine screening method |
| CN104951870A (en) * | 2015-06-01 | 2015-09-30 | 南通科瑞斯生物医药科技有限公司 | Pharmaceutical pre-clinical cardiac risk assessment method |
| CN107271798A (en) * | 2016-04-08 | 2017-10-20 | 首都医科大学 | A kind of monophasic action potential record system |
-
2018
- 2018-12-05 CN CN201811478355.6A patent/CN109765348A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU1790768C (en) * | 1989-09-13 | 1993-01-23 | Л.В.Бобровников | Method of nerve cell membrane permeability determination |
| CN102883653A (en) * | 2010-03-22 | 2013-01-16 | 莱斯特大学 | Method and apparatus for evaluating cardiac function |
| CN104237308A (en) * | 2013-06-09 | 2014-12-24 | 国家纳米科学中心 | In-vitro medicine screening method |
| CN104951870A (en) * | 2015-06-01 | 2015-09-30 | 南通科瑞斯生物医药科技有限公司 | Pharmaceutical pre-clinical cardiac risk assessment method |
| CN107271798A (en) * | 2016-04-08 | 2017-10-20 | 首都医科大学 | A kind of monophasic action potential record system |
Non-Patent Citations (5)
| Title |
|---|
| 张永健等: "双苯氟嗪对人心房肌纤维电生理的影响(英文) ", 《中国药理学与毒理学杂志》 * |
| 张鲲等: "昂丹司琼致QT间期延长的研究进展", 《中国新药杂志》 * |
| 徐惠等: "SOTALOL对新生犬心肌浦肯野纤维电生理特性的影响 ", 《中国医科大学学报》 * |
| 郭舜等: "HERG钾离子通道与药物心脏毒性的安全性评价 ", 《医学综述》 * |
| 陈龙等: "ICH对药物临床前心脏安全性评价的技术要求 ", 《中国新药杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109856349A (en) * | 2018-12-05 | 2019-06-07 | 南通科瑞斯生物医药科技有限公司 | A kind of optimization method of high throughput hERG assessment cardiac safety risk |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | A cardiomyocyte-based biosensor for antiarrhythmic drug evaluation by simultaneously monitoring cell growth and beating | |
| Farias et al. | Definition of reference ranges for the platelet distribution width (PDW): a local need | |
| Robinson et al. | Rapid dopamine release in freely moving rats | |
| WO2007066684A1 (en) | Method of measuring cell motility and system therefor | |
| Booth et al. | Brain white matter tract integrity and cognitive abilities in community-dwelling older people: the Lothian Birth Cohort, 1936. | |
| CN114002421B (en) | Application of exosome metabolite as bipolar affective disorder marker | |
| JP5556144B2 (en) | Blood cell analyzer, blood cell analysis method, and blood cell analysis system | |
| US9448228B2 (en) | Cell analyzing apparatus and cell analyzing method | |
| CN103405225A (en) | Method, apparatus and device for obtaining pain feeling evaluation indexes | |
| EP3825682B1 (en) | Herpesvirus measuring method and herpes virus measuring device | |
| CN109765348A (en) | A kind of accurately external medicine heart peace comments method | |
| KR102257995B1 (en) | In vitro diagnostic apparatus and method for in vitro diagnostic test thereof | |
| Leal et al. | Determination of schizophrenia using electronic nose via support vector machine | |
| CN108445200A (en) | A kind of influence based on micro-fluidic chip detection pentoxifylline to Erythrocytes from Coronary Heart Disease deformability and biochemical index | |
| KR20190074422A (en) | Blood analyzer and method for blood analysis using the same | |
| US20200209218A1 (en) | Alarming method for platelet aggregation sample, blood cell analyzer and storage medium | |
| JP2006521550A (en) | Method and system for analyzing compounds | |
| DE60028374T2 (en) | METHOD AND APPARATUS FOR PRESENTING DATA OF THROMBOSIS AND HAEMOSTASE ASSAYS | |
| CN111951964A (en) | Method and system for rapidly detecting novel coronavirus pneumonia | |
| Torlakovic | How to validate predictive immunohistochemistry testing in pathology? | |
| Liu | Electrophysiological studies of voltage‐gated sodium channels using QPatch HT, an automated patch‐clamp system | |
| US20060064028A1 (en) | Method for drawing up data which can be used to assess cognitive or sensomotor capabilities or capacities of people subjected to a test | |
| CN106990232B (en) | Blood Kit analysis system and analysis method | |
| Ramlugun et al. | A comprehensive framework for evaluation of high pacing frequency and arrhythmic optical mapping signals | |
| FI3658884T3 (en) | Method for immobilising biological samples for analytical and diagnostic purposes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190517 |