CN103565783B - The application in voltage gated k+ channel blocker agonist prepared by hexachlorophene - Google Patents
The application in voltage gated k+ channel blocker agonist prepared by hexachlorophene Download PDFInfo
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Abstract
The invention provides the application of a kind of hexachlorophene in preparation KCNQ agonist; In addition, present invention also offers a kind of hexachlorophene in preparation as the application in the medicine of KCNQ agonist; Wherein, in above-mentioned application, described KCNQ agonist can be KCNQ1 agonist, KCNQ1/KCNE1 agonist, KCNQ2 agonist or KCNQ2/KCNQ3 agonist.Present invention show hexachlorophene and there is enhancing KCNQ1/KCNE1 electric current, promote Single Cardiac Cell repolarization, reach potential duration under reach, treat ARR effect; Hypotype potentiation shows, hexachlorophene strengthens KCNQ and M electric current, by strengthening KCNQ2 electric current and KCNQ2/KCNQ3 different aggressiveness Current Regulation level of membrane potential, weaken the effect that action potential provides frequency, achieve the application of hexachlorophene in the medicine being epilepsy target spot is treated with KCNQ in preparation, reach antiepileptic efficacy.
Description
Technical field
The invention belongs to drug world, particularly, the present invention relates to the application of hexachlorophene in preparation KCNQ agonist, wherein, in above-mentioned application, described KCNQ agonist can be KCNQ1 agonist, KCNQ1/KCNE1 agonist, KCNQ2 agonist or KCNQ2/KCNQ3 agonist.In addition, present invention also offers a kind of hexachlorophene in preparation as the application in the medicine of KCNQ agonist, wherein, in above-mentioned application, described KCNQ agonist can be KCNQ1 agonist, KCNQ1/KCNE1 agonist, KCNQ2 agonist or KCNQ2/KCNQ3 agonist.In addition, the invention still further relates to hexachlorophene and prepare the application in antiarrhythmic drug and antiepileptic.
Background technology
Hexachlorophene (Hexachlorophene, HCP), also referred to as Nabac, is widely used as the antiinflammatory germ killing drugs of clinical local.Chemistry full name: 2,2 '-di-2-ethylhexylphosphine oxide (3,4,6-trichlorophenol, 2,4,6,-T), chemical constitution is:
Hexachlorophene, as a kind of prescription drugs, is a kind of strong effective antibacterial skin antiseptic (Jungermann, E.JAmOilChemSoc1968,45 (5), 345-50) being widely used as treatment acne.Previous research shows, hexachlorophene plays antitumaous effect (Park, S. etc., MolPharmacol2006,70 (3), 960-6.) by suppressing Wnt/beta-catenin signaling pathway.But up to the present, about the research report on hexachlorophene effect ion channel is less.
Ion channel be positioned at a class on cytoplasma membrane can the controlled albumen of quick penetrating ion, being distributed widely in each histoorgan of human body, playing critical function in vital movement, is an important target of drug effect and research and development.Potassium-channel be distribute the most extensively, ion channel that quantity is maximum, and voltage gated k+ channel blocker is a wherein important class, has 12 member (K
v1.X-K
v12.X).KCNQ passage is the 7th member (Kv7) of voltage-gated potassium channels, comprises five hypotype KCNQ1-KCNQ5.Compared with other potassium channel, KCNQ passage activation threshold is lower, under action potential threshold value, (-60mV) can open, and it activates slowly, the also non-inactivation when LLD, these features make KCNQ passage in promotion cell repolarization, regulate in nervous cell membrane potential level and play an important role.As same family member, except the homology had in sequence, KCNQ hypotype passage is all made up of four subunits, and each subunit comprises 6 transmembrane segments (S1-S6), wherein S1-S4 forms the voltage receptive field of passage, and S5-S6 forms the structural concordance such as the duct district of passage.But as different subtype, KCNQ passage hypotype is at current characteristic or in distribution, physiological function and formed on complex with other hypotypes and all have different characteristic, they have determined that the multiformity of KCNQ drug target.Therefore KCNQ member had both had similarity and had also had specificity.
KCNQ1 is mainly distributed in cardiac system, can not form different aggressiveness passage with other hypotype of KCNQ, only forms the I in myocardial action potential with complementary subunit KCNE1
kselectric current, is responsible for myocardial action potential rapid repolarization three phase, affects myocardial action potential time-histories.KCNQ2-KCNQ5 is mainly distributed in nervous system, and KCNQ2/KCNQ3 is the main composition of composition M electric current, has report to show that KCNQ5 also participates in the formation of M electric current, M Current Regulation membrane potential of neurons level.KCNQ4 is mainly arranged in internal ear, participates in the formation (JentschT.J.NatRevNeurosci2000,1,21-30.) of audition.
KCNQ passage is in adjustment cardiomotility, and the different aggressiveness passage KCNQ1/KCNE1 be made up of KCNQ1 subunit complementary with it KCNE1 completes.The action potential of a complete ventricular muscle cell comprises depolarization, 1 phase/rapid repolarization initial stage, 2 phases/plateau, latter stage 3 phases/rapid repolarization, 4 phases/quiescent stage, and repolarization 2 phase and 3 phases determine action potential length, have the build-in attribute of refractory stage at myocardial cell under, the prolongation of action potential duration, APD is by cardiac arrhythmia that to cause with surface electrocardiogram QT interval prolongation be feature.Be responsible for the export-oriented I of 2 repolarization latter stages phase and 3 phase repolarizations mainly KCNQ1/KCNE1 passage composition
kselectric current, I after KCNQ1 sudden change
kscurrent decay causes a type long QT syndrome, is the main pathogenic factor of long QT syndrome.
KCNQ passage, in adjustment nervous cell membrane potential level, is mainly completed by the M electric current be distributed in nervous system.The process of depolarization of neurocyte have activated M channel current, in born of the same parents, potassium concentration is higher than under the ion basic condition outside born of the same parents, positively charged efflux of K+ ions, transmembrane potential is thus by just becoming negative, cell excitability reduces, thus avoid neuronic seriality, epileptiform discharge that excessive synchronous discharge is brought out.Therefore, primarily of the M electric current of KCNQ passage composition in maintenance neural cell excitability balance, in the nerve excitability diseases such as the epilepsy that causes of avoiding being lacked of proper care by irritability, play important physiological significance.
This research and utilization whole cell protein electrophoresis, the technological means such as binding molecule biology, find that hexachlorophene can strengthen the different aggressiveness channel current of KCNQ1/KCNE1, potential duration under reach, reach antiarrhythmic effect, hypotype effect shows that hexachlorophene strengthens KCNQ passage, shows it and can be used as antiepileptic composition or the drug development as parent nucleus.
Summary of the invention
The invention provides the application of a kind of hexachlorophene in preparation KCNQ agonist;
Wherein, in above-mentioned application, described KCNQ agonist can be KCNQ1 agonist, KCNQ1/KCNE1 agonist, KCNQ2 agonist or KCNQ2/KCNQ3 agonist.
In addition, present invention also offers a kind of hexachlorophene in preparation as the application in the medicine of KCNQ agonist;
Wherein, in above-mentioned application, described KCNQ agonist can be KCNQ1 agonist or KCNQ1/KCNE1 agonist; Wherein, described medicine is used for the treatment of and/or prevents arrhythmia or long QT syndrome;
Further, in above-mentioned application, described KCNQ agonist can be KCNQ2 or KCNQ2/KCNQ3 agonist; Wherein, described medicine is used for the treatment of and/or prevents epilepsy.
Present invention show hexachlorophene and there is enhancing KCNQ1/KCNE1 electric current, promote Single Cardiac Cell repolarization, reach potential duration under reach, treat ARR effect; Hypotype potentiation shows, hexachlorophene strengthens KCNQ and M electric current, by strengthening KCNQ2 electric current and KCNQ2/KCNQ3 different aggressiveness Current Regulation level of membrane potential, weaken the effect that action potential provides frequency, achieve the application of hexachlorophene in the medicine being epilepsy target spot is treated with KCNQ in preparation, reach antiepileptic efficacy.
Accompanying drawing explanation
Fig. 1 is the impact of hexachlorophene on KCNQ1 and KCNQ1/KCNE1 electric current, wherein:
Figure 1A is map of current before and after KCNQ1 passage administration 10 μMs of hexachlorophene;
Figure 1B is map of current before and after KCNQ1/KCNE1 different aggressiveness passage administration 10 μMs of hexachlorophene;
Fig. 1 C is conductance voltage curve before and after KCNQ1 passage administration 10 μMs;
Fig. 1 D is conductance voltage curve before and after KCNQ1/KCNE1 different aggressiveness passage administration 10 μMs of hexachlorophene;
Fig. 1 E is that the lower 10 μMs of hexachlorophene of-10mV test voltage strengthen KCNQ1 and KCNQ1/KCNE1 channel current cartogram;
Fig. 1 F is the dose effect curve of the hexachlorophene of the different aggressiveness passage of KCNQ1/KCNE1.
Fig. 2 is the impact of hexachlorophene on Single Cardiac Cell time-histories, wherein:
Fig. 2 A shows 1 μM of hexachlorophene and 3 μMs of hexachlorophene to the impact of the adult guinea pig left compartment muscle action potentials of cells time-histories of acute isolation;
Fig. 2 B is that 3 μMs of hexachlorophene are on Single Cardiac Cell time-histories, APD90(action potential multipole 90% time-histories) and APD50(action potential half repolarization duration) affect cartogram.
Fig. 3 is the impact of hexachlorophene on a type long QT syndrome, wherein:
Fig. 3 A is map of current before and after KCNQ1 sudden change R243C passage administration 10 μMs of hexachlorophene;
Fig. 3 B is map of current before and after KCNQ1 sudden change R539W passage administration 10 μMs of hexachlorophene;
Fig. 3 C is map of current before and after KCNQ1 sudden change R243C/KCNE1 different aggressiveness passage administration 10 μMs of hexachlorophene;
Fig. 3 D is map of current before and after KCNQ1 sudden change R539W/KCNE1 different aggressiveness passage administration 10 μMs of hexachlorophene;
Fig. 3 E is conductance voltage curve before and after KCNQ1 sudden change R243C passage administration 10 μMs of hexachlorophene;
Fig. 3 F is conductance voltage curve before and after KCNQ1 sudden change R539W passage administration 10 μMs of hexachlorophene;
Fig. 3 G is conductance voltage curve before and after KCNQ1 sudden change R243C/KCNE1 different aggressiveness passage administration 10 μMs of hexachlorophene;
Fig. 3 H is conductance voltage curve before and after KCNQ1 sudden change R539W/KCNE1 different aggressiveness passage administration 10 μMs of hexachlorophene.
Fig. 4 is the impact of hexachlorophene on KCNQ passage hypotype electric current, wherein:
Fig. 4 A is map of current before and after KCNQ2 passage administration 10 μMs of hexachlorophene;
Fig. 4 B is map of current before and after KCNQ2/KCNQ3 different aggressiveness passage administration 10 μMs of hexachlorophene;
Fig. 4 C is map of current before and after KCNQ4 passage administration 10 μMs of hexachlorophene;
Fig. 4 D is conductance voltage curve before and after KCNQ2 passage administration 10 μMs of hexachlorophene;
Fig. 4 E is conductance voltage curve before and after KCNQ2/KCNQ3 different aggressiveness passage administration 10 μMs of hexachlorophene.
Detailed description of the invention
Specific embodiments of the invention are described in detail below in conjunction with technical scheme and accompanying drawing.
Embodiment 1 hexachlorophene strengthens the different aggressiveness channel current of KCNQ1 and KCNQ1/KCNE1
1. cell culture and transfection
Chinese hamster ovary cell (ChineseHamsterOvary, CHO) (Chinese Academy of Sciences's cell bank) culture fluid formula: 50/50DMEM/F-12(Gibco), add 10% hyclone (Fetalbovineserum, FBS) (Gibco, Australia), 2mML-glutamine (Invitrogen).
(KCNQ1 plasmid is by Zentrumf ü rMolekulareNeurobiologie for expressing K CNQ1 passage, professor ThomasJ.Jenstsch of Hamburg is so kind as to give, sequence is through gene sequencing and utilize Megalign7(DNAStar) sequence analysis comparison software prove with ncbi database (NationalCenterofBiotechnologyInformation, NCBI, US National Biotechnology Information center) in numbering NM_000218.2(KCNQ1) sequence is consistent): first 24 hours of transfection, use trypsin Sigma, China) digestion after be laid in the ware of diameter 35mm.Transfection uses Lipofectamine2000
tMreagent (Invitrogen), operates according to its operating procedure provided (protocol).After transfection 24 hours, be again laid on after cell dissociation on slide that poly-L-Lysine (poly-L-lysine) (Sigma) soak.Cotransfection GFP(green fluorescent protein), transfectional cell can be confirmed under fluorescence microscope.
Expressing K CNQ1/KCNE1(KCNE1 plasmid is so kind as to give by professor M.C.Sanguinetti of University of Utah (UniversityofUtah), GaoZ.G. etc., JBiolChem2008,283 (33), 22649-58.) time, KCNQ1/KCNE1 presses plasmid mass ratio 50/50 transfection, and step and method is identical with expressing K CNQ1 passage.
Electrophysiological recording on 2.CHO cell
At room temperature, Axopatch-700B amplifier (MolecularDevices, Sunnyvale, CA) is adopted to carry out full cell type voltage-clamp recording.Borosilicate glass capillary tube (WorldPrecisionInstrunents, Sarasota, FL) is drawn into electrode, and the resistance in electrode in tytosis after liquid is 3-5M Ω.
Cell Inner formula of liquid: 145mMKCl, 1mMMgCl
2, 5mMEGTA, 10mMHEPES, 5mMMgATP(KOH are adjusted to pH=7.3).During record, by BPS perfusion system (ALAScientificInstruments, Westburg, NY) continous perfusion extracellular fluid.
Extracellular fluid formula: 140mMNaCl, 5mMKCl, 2mMCaCl
2, 1.5mMMgCl
2, 10mMHEPES, 10mM glucose (NaOH is adjusted to pH=7.4).In pClamp9.2 software (MolecularDevices, Sunnyvale, CA), digital signal is converted to DigiData1322A after signal of telecommunication 2kHz filtering.Series resistance compensation 60-80%, adopts multivoltage scheme in electrophysiologic study recording process.
3. hexachlorophene strengthens KCNQ1 and KCNQ1/KCNE1 channel current
Weigh 11.6mg hexachlorophene (Sigma company, analytical pure) to be dissolved in 285 μ lDMSO (Sigma company, D5879, >=99.5%), obtain 100mM hexachlorophene mother solution, in-20 DEG C of cold preservations.Get 10 μ l in experimentation in 20ml extracellular fluid, and supplement 10 μ lDMSO with controls administration solution D MSO percent by volume to make a gesture of measuring for DMSO volume basis in all test concentrations of 1%(be 1%), mix, reach 10 μMs of hexachlorophene test concentrations.
In electrophysiological recording, KCNQ1 channel current record scheme is: Clamping voltages-80mV, a series of 1500ms stimulation voltage (from-80mV to+50mV, interval 10mV) induced current, induces tail current by going through 1500ms-120mV.KCNQ1 passage to be expressed electric current I under-10mV test voltage
0after stable, utilize BPS perfusion system to 10 μMs of hexachlorophene, until electric current I is stablized.
KCNQ1/KCNE1 channel voltage pincers record scheme is: Clamping voltages-110mV, and a series of 3000ms stimulation voltage (from-80mV to+80mV, interval 10mV) induced current induces tail current by going through 2500ms-120mV.KCNQ1 passage to be expressed electric current I under-10mV test voltage
0after stable, utilize BPS perfusion system to 10 μMs of hexachlorophene, until electric current I is stablized.
4. experimental result statistical analysis:
Electric physiological data analysis and utilization Clampfit10.2(MolecularDevices, Sunnyvale, CA), recycling GraphPadPrim5(GraphPadSoftware, SanDiego, CA), Boltzmann's equation (Boltzmannequation) matching of voltage independent activating curve, dose effect curve Xi Er equation (Hillequation) matching.Obtain V
1/2for half activation voltage, Δ V
1/2for half activation voltage moves to left value, negative sign (-) represents that current activation curve moves to left, and positive sign (+) represents and moves to right, I/I
0for activating multiple.
As shown in Figure 1,10 μMs of hexachlorophene are directly acted on the different aggressiveness passage of KCNQ1 and KCNQ1/KCNE1 of expressing on the gonad cell of Chinese hamster, under-10mV test voltage, before administration, channel current strengthens 1.48 ± 0.13 (Fig. 1 .A and E) and 4.70 ± 0.70 times (Fig. 1 .B and E) respectively.Conductance voltage curve (G-Vcurve) is the common counter of reaction channel degree of opening, after administration 10 μMs of hexachlorophene, and the Δ V of the different aggressiveness of KCNQ1 and KCNQ1/KCNE1
1/2move to left-29.92 ± 1.93mV and-66.05 ± 6.15mV respectively, display hexachlorophene can promote the opening (Fig. 1 .C and D) of passage, Ion channel kinetics statistics shows, hexachlorophene obviously delays the closedown (p < 0.0001) of KCNQ1 passage, shows that hexachlorophene plays by delaying pathway closure and promotion channel opener etc. the effect strengthening current intensity.Dose-dependent relationship is the important indicator whether compound of reaction has pharmacotoxicological effect, and as shown in Fig. 1 .F, KCNQ1/KCNE1 different aggressiveness passage has obvious dose-dependence to hexachlorophene, its EC50=4.95 ± 0.09 μM.Above data show, hexachlorophene is the agonist of the different aggressiveness passage of a kind of novel KCNQ1 and KCNQ1/KCNE1.
Embodiment 2 hexachlorophene shortens Single Cardiac Cell time-histories
In normal myocardial cell, KCNQ1/KCNE1 different aggressiveness passage forms export-oriented I
kselectric current participates in 2 phases of Single Cardiac Cell/latter stage plateau and 3 phases/rapid repolarization, has an impact to Single Cardiac Cell time-histories.But, not yet unified understanding is had to KCNQ1 and the KCNE1 formation ratio at body due to current, think that can the hexachlorophene investigated having potentiation at Chinese hamster ovary cell by the KCNQ1/KCNE1 different aggressiveness passage of 50/50 mass ratio heterogenous expression produce potentiation at the different aggressiveness passage of the KCNQ1/KCNE1 of body, can by strengthening KCNQ1/KCNE1 different aggressiveness passage and then affecting whole Single Cardiac Cell time-histories to investigate hexachlorophene, 1 μM and 3 μMs of hexachlorophene are acted on the guinea pig left ventricular myocyte of acute isolation by the present inventor respectively, by current clamp hands section, the present inventor have studied the impact of hexachlorophene on Single Cardiac Cell time-histories.
Concrete grammar adopts the Langerdorff perfusion system condition (AkaoM. etc. of previously report, CircRes2001,88,1267-75.) under, acute isolation adult guinea pig myocardium of left ventricle cell, concrete steps are: cut left ventricle rapidly along guinea pig heart center line, with rinsing 5-6 minute containing the collagenase of 6mg/ml and the protease of 0.1mg/ml without calcium tyrode's solution (Tyrode ' ssoultion); Use KB liquid (Kraft-Bruhesolution) to rinse 5 minutes again, shred ventricular organization, 4 DEG C of cold preservations are stand-by.KB formula of liquid: 50mML-glutamic acid, 80mMKOH, 40mMKCl, 3mMMgSO
4, 25mMKH
2pO
4, 10mMHEPES, 1mMEGTA, 20mM taurine, 10mM glucose (KOH is adjusted to pH=7.4).Myocardial action potential is recorded as and utilizes Axonpatch-200B amplifier (MolecularDevices, Sunnyvale, CA) carry out full cell type current clamp record, ventricular muscles current clamp record scheme is: give 10ms depolarization impulse stimulation with 0.16HZ frequency, operation of recording potential amplitude.
Weigh hexachlorophene (Sigma company, analytical pure) to be dissolved in extracellular fluid to 1 μM and 3 μMs of test concentrations.Cell Inner formula of liquid: 120mM potassium aspartate, 20mMKCl, 1mMMgSO
4, 4mMNa2ATP, 10mMHEPES(KOH are adjusted to pH=7.2); Extracellular fluid formula: 135mMNaCl, 5.4mMKCl, 1.8mMCaCl
2, 1mMMgCl
2, 5mMHEPES, 10mM glucose (NaOH is adjusted to pH=7.4).
Experimental result shows, 1 μM of hexachlorophene potential duration under reach, and shortening effect is faint, does not have statistical significance; But 3 μMs of hexachlorophene significance potential duration under reach (Fig. 2 .A), illustrate as in experiment in vitro, the KCNQ1/KCNE1 of heterogenous expression is similar, hexachlorophene can dose-dependant ground potential duration under reach as agonist, but on action potential morphology without impact.Statistical result shows, after administration 3 μMs of hexachlorophene, guinea pig cardiomyocytes action potential duration, APD APD90(action potential multipole 90% time-histories) and APD50(action potential half repolarization duration) shorten respectively 50.7 ± 9.8% and 53.1 ± 9.1%(Fig. 2 .B).Above experimental data shows, hexachlorophene, as a kind of KCNQ1/KCNE1 agonist, can shorten at body-centered myocyte's action potential duration, APD by significance under low dosage condition, can be used for preparing in the medicine as KCNQ1/KCNE1 agonist.
A type long QT syndrome mutant channel function saved by embodiment 3 hexachlorophene
The formation of a complete Single Cardiac Cell, different ion channels is participated, and comprises KCNQ1, HERG, the SCN5A (AmJPhysiolHeartCircPhysiol2000 such as Bianchi, L., 279 (6), H3003-11. and the HumMutat2009 such as Hedley, P.L., 30 (11), 1486-511.), KCNE1, KCNE2 etc., the exception of these ion channel function all will cause long QT syndrome.But, suddenly change in the long QT syndrome that causes at these, be suddenlyd change caused by the miopragia that causes by KCNQ1 more than the acquired long QT syndrome of 50% congenital long QT syndrome and more than 90%, belong to a type long QT syndrome.
Vivo and vitro experiment has shown that hexachlorophene is as a kind of KCNQ1 and KCNQ1/KCNE1 agonist, can strengthen channel current, and move to left conductance voltage curve, the pharmacotoxicological effects such as potential duration under reach.Whether there is for detecting hexachlorophene the KCNQ1 mutation current of enhancing one type long QT syndrome, as rescue long QT syndrome agonist, being applied in antiarrhythmic drug.The present inventor's random selecting four causes the KCNQ1 of a type long QT syndrome to suddenly change, by QuickChange II point mutation software (Stratagene, LaJolla, CA) primer is built, increased by PfuUltraII thermal starting archaeal dna polymerase (Strategene), utilize gene sequencing and Megalign7(DNAStar) carry out sequence alignment checking sudden change successfully construct after, sudden change KCNQ1 passage and the sudden change KCNQ1/KCNE1 passage of external heterogenous expression in Chinese hamster ovary cell is recorded in by voltage clamp technology, channel protein is expressed, electrophysiological recording and data analysis concrete operation method step identical with embodiment one.
Four sport KCNQ1R190Q, T587M, R243C, R539W, wherein because electric current in the first two sudden change electrophysiological recording process is too small, do not give administration; Hexachlorophene all has agonism to R243C and R539W.As shown in Figure 3, under-10mV test voltage, before administration 10 μMs of hexachlorophene, KCNQ1 sudden change R243C and R539W channel current strengthens 1.54 ± 0.26 and 1.84 ± 0.21 times (Fig. 3 .A and B) respectively; Move to left conductance voltage curve, Δ V
1/2move to left-22.41 ± 1.03mV and-30.83 ± 0.55mV(Fig. 3 .E and F respectively).It is 1.44 ± 0.33 and 2.91 ± 0.89 times (Fig. 3 .C and D) respectively that 10 μMs of hexachlorophene strengthen the different aggressiveness channel current of KCNQ sudden change R243C/KCNE1 and R539W/KCNE1; The conductance voltage curve of the different aggressiveness passage of the R539W/KCNE1 that moves to left, Δ V
1/2=-29.35 ± 2.74 (Fig. 3 .H).The conductance voltage curve of hexachlorophene to the different aggressiveness passage of R243C/KCNE1 can be divided into two, and move to left under low-voltage curve, and move to right under high voltage curve, and this explains why 10 μMs of hexachlorophene can the different aggressiveness channel current of exciting sudden change R243C/KCNE1, and Δ V
1/2=+3.9 ± 4.0(Fig. 3 .G).Above experimental data shows, hexachlorophene has the pharmacological effects saved and cause the KCNQ1 of a type long QT syndrome to suddenly change, and shows that hexachlorophene can be used for preparing the antiarrhythmic drug as KCNQ1/KCNE1 agonist further.
Embodiment 4 hexachlorophene affects KCNQ passage hypotype
Finding that under the background that hexachlorophene has agonism to KCNQ1, the present inventor continues to have investigated the impact of hexachlorophene to KCNQ2, KCNQ2/KCNQ3 and KCNQ4 passage.Wherein the M channel current of KCNQ2/KCNQ3 composition is the main component regulating level of membrane potential, and channel function weakens and will neural cell excitability be caused to lack of proper care, and final initiation takes place frequently as the epileptic condition of feature with action potential.KCNQ4 is mainly arranged in internal ear, participates in the formation of audition, and miopragia after sudden change, causes the non-syndrome sensorineural hearing loss (NonsyndromicSensorineuralDeafnesstype2, DFNA2) of 2 type.
KCNQ2 plasmid is by the New York State University (StateUniversityofNewYork, StonyBrook) professor D.Makinnon is so kind as to give, KCNQ3 plasmid is so kind as to give by professor M.C.Sanguinetti of University of Utah (UniversityofUtah), KCNQ4 is by health science center, Texas (UniversityofTexasHealthScienceCenter, SanAntonio) professor M.Shapiro is so kind as to give, above sequence is through gene sequencing and utilize Megalign7(DNAStar) sequence analysis comparison software proves, respectively with ncbi database in announce be numbered NM_133322.1(KCNQ2), NM_004519.3(KCNQ3), NM_004700.3(KCNQ4) sequence is consistent, the expression of passage, record and data analysing method step are with embodiment one.
Under-10mV test voltage, before administration 10 μMs of hexachlorophene, KCNQ2, KCNQ2/KCNQ3 and KCNQ4 channel current strengthens 1.65 ± 0.22,1.25 ± 0.13 and 6.87 ± 2.27 times (Fig. 4 .A, B and C) respectively.In the impact on conductance voltage, after administration 10 μMs of hexachlorophene, except KCNQ4 passage cannot be added up, the Δ V of KCNQ2 and KCNQ2/KCNQ3 passage
1/2move to left-40.05 ± 1.91mV and-27.67 ± 2.13mV(Fig. 4 .D and E respectively).Above experimental data shows, hexachlorophene except can be used as KCNQ1 and KCNQ1/KCNE1 agonist and as except the application in the anti-depressant medications of target spot, also there is the effect strengthening KCNQ2 and KCNQ2/KCNQ3 passage, show that hexachlorophene can have epilepsy disease effects; Strengthen the effect of KCNQ4 channel current, show that hexachlorophene can be used for treating and/or preventing relevant disease as in the non-syndrome sensorineural hearing loss of 2 type or model.
As indicated in the present invention, hexachlorophene can strengthen the different aggressiveness channel current of KCNQ1 and KCNQ1/KCNE1, shows the agonist that it is the different aggressiveness passage of a kind of novel KCNQ1 and KCNQ1/KCNE1; Meanwhile, hexachlorophene, as a kind of KCNQ1/KCNE1 agonist, can shorten at body-centered myocyte's action potential duration, APD by significance under low dosage condition, can be used for preparing in the medicine as KCNQ1/KCNE1 agonist; In addition, found by the KCNQ1/KCNE1 sudden change suddenlyd change to the type long QT interval expressed, hexachlorophene can save the current attenuation suddenling change and cause, show it and there is the pharmacological effects saved and cause the KCNQ1 of a type long QT syndrome to suddenly change, can be used for preparing the antiarrhythmic drug as KCNQ1/KCNE1 agonist; In addition, use in passage hypotype, show hexachlorophene except can be used as KCNQ1 and KCNQ1/KCNE1 agonist and as except the application in the anti-depressant medications of target spot, also there is the effect strengthening KCNQ2 and KCNQ2/KCNQ3 passage, show it and can have epilepsy disease effects; Hexachlorophene also has the effect of enhancing KCNQ4 channel current simultaneously, shows it and can be used for treating and/or preventing relevant disease as in the non-syndrome sensorineural hearing loss of 2 type or model.
Claims (8)
1. the application of hexachlorophene in preparation KCNQ agonist.
2. application according to claim 1, wherein, described KCNQ agonist is KCNQ1 agonist, KCNQ1/KCNE1 agonist, KCNQ2 agonist or KCNQ2/KCNQ3 agonist.
3. a hexachlorophene is being prepared as the application in the medicine of KCNQ agonist.
4. application according to claim 3, wherein, described KCNQ agonist is KCNQ1 agonist or KCNQ1/KCNE1 agonist.
5. application according to claim 4, wherein, described medicine is used for the treatment of and/or prevents long QT syndrome.
6. application according to claim 4, wherein, described medicine is used for the treatment of and/or prevents arrhythmia.
7. application according to claim 3, wherein, described KCNQ agonist is KCNQ2 or KCNQ2/KCNQ3 agonist.
8. application according to claim 7, wherein, described medicine is used for the treatment of and/or prevents epilepsy.
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| CN102018687A (en) * | 2010-11-24 | 2011-04-20 | 广东省农业科学院兽医研究所 | Application of hexachlorophene in resistance to Eimeria tenella |
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| Hexachlorophene Is a Potent KCNQ1/KCNE1 Potassium Channel Activator Which Rescues LQTs Mutants;Yueming Zheng等;《PLOS ONE》;20121212;第7卷(第12期);第1-9页 * |
| 六氯酚粉与新生儿葡萄球菌感染;Allen KD 等;《中国消毒学杂志》;19951231;第12卷(第1期);第61页 * |
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