CN1306037C - Novel use of cardiocyte M3 receptor and IkM3 in screening medicament for treating arrhythmia - Google Patents
Novel use of cardiocyte M3 receptor and IkM3 in screening medicament for treating arrhythmia Download PDFInfo
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- CN1306037C CN1306037C CNB2004100439458A CN200410043945A CN1306037C CN 1306037 C CN1306037 C CN 1306037C CN B2004100439458 A CNB2004100439458 A CN B2004100439458A CN 200410043945 A CN200410043945 A CN 200410043945A CN 1306037 C CN1306037 C CN 1306037C
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Abstract
The present invention provides a new use of a myocardial cell M3 receptor and IKM3 for screening arrhythmia treating drugs, which relates to a new use of myocardial cells. At present, people can not find a definite effective standard for arrhythmia treating drugs, and the present invention solves the problem. A method of screening drugs using the myocardial cell M3 receptor and the IKM3 as targets has the advantages of high accuracy, strong specificity and easy operation. The screened drugs thereby have high specificity and obviously reduce the adverse reactions of the existing antiarrhythmic drugs to enhance curative effect.
Description
Technical field:
The present invention relates to a kind of myocardial cell's new purposes, specifically with myocardial cell's muscarinic acetylcholine receptor M3 hypotype and IK
M3Electric current is the purposes of the screening antiarrhythmic drug of target.
Background technology:
Irregular pulse serious harm human health is to cause one of human main causes of death, and epidemiological study shows that 50% heart failure patient is died from irregular pulse.Although some antiarrhythmic drugs improve conditions of patients, but still all exist obviously not enough: 1. antiarrhythmic drug exists total effective rate not high, only is 30%～60%; 2. invalid to some obstinate irregular pulse; 3. prolonged application is not good to patient's survival rate improvement effect; 4. be easy to occur problems such as proarrhythmia side effect.And, before not having practical application, people are difficult to judge which kind of medicine has the ARR effect of treatment, when screening treatment antiarrhythmic medicament, people also can not find definite, an effective standard, and the development of high-efficiency low-toxicity Antiarrhythmic Agent is extremely urgent.
The core of modern new drug development is to be designated as the basis with drug target, and filtering out on this basis can excitement or the biological function of antagonism target mediation, has the high-efficiency low-toxicity medicine or the lead compound of therapeutic action.Drug targets is the biomacromolecule material that plays a crucial role in the disease development process, as acceptor, ionic channel, proteolytic enzyme etc.Heart muscarinic acetylcholine receptor (hereinafter to be referred as m receptor) has vital role in regulating cardiac electrical activity and myocardial contraction.Have been found that at present and restrain and fall 5 kinds of hypotypes of m receptor, respectively called after M
1-M
5But it is believed that for a long time only there is M in heart
2Hypotype.The inventor finds exciting heart M at first under study for action
3Acceptor and IK
M3Can mediate a kind of new potassium current, first called after IK
M3(Shi H, Wang H, Lu Y, Yang B, Wang Z.Choline modulates cardiacmembrane repolarization by activating an M3 muscarinic receptor and its coupled K+channel.J Membr Biol.1999; 169 (1): 55-64), and find to have M in the human heart
3Receptor mrna and protein expression confirm to have M the heart from function assessment and molecular biology angle
3Acceptor (Baofeng Yang Zhiguo Wang Cardiac M3 receptor:to be or not to be, that is thequestion Recent Res.Devel.Life Sci.2003; 1:73-95).Further discover M
3Acceptor has physiological functions such as hyperpolarization membrane potential, current potential time-histories under reach, reducing heart rate, and (the precious peak of Liu Yan, king's Yue, Ma Manling, Zhang Yan, Li Houwei, Chen Qingwen, poplar M3 receptor stimulant is to the Acta Pharmaceutica Sinica that influences of rat and rabbit cardiac hemodynamic, 2001; 36 (2): 84-87), what is more important is found heart M
3Acceptor and irregular pulse and apoptosis of cardiac muscle have confidential relation (the precious peak M of the magnificent Li Hu human relations of Liu Yan chaste tree rubine grandson poplar
3The research Acta Pharmaceutica Sinica 2004,39 (5) of acceptor and rat ischemia apoptosis of cardiac muscle relation; 338-341), but up to now, still do not know heart M
3Acceptor and IK
M3Functionalized use.
Summary of the invention:
The present invention be exactly at people when the screening treatment antiarrhythmic medicament owing to lack effective drug targets to making the medicine that can't filter out high-efficiency low-toxicity, the object of the present invention is to provide a kind of myocardial cell M that can practical application
3Acceptor and IK
M3As the purposes of screening treatment antiarrhythmic medicament, its feature is myocardial cell M
3Acceptor and IK
M3Purposes with screening treatment antiarrhythmic medicament: add medicine to be screened in the extracellular fluid, write down strength of current under the full cell pattern, strength of current then the ARR medicine of treatment that descends before than medication under 50mv voltage for having curative effect.According to myocardial cell M
3Acceptor and IK
M3For target carries out the method for screening of medicaments, has the accuracy height, high specificity, easy handling advantage.Utilize myocardial cell M
3Acceptor and IK
M3The medicine of screening not only has the height specificity, and will obviously reduce the untoward reaction of present antiarrhythmic drug, improves curative effect of medication.Inventor's experimental study shows, acts on M
3Acceptor and IK
M3Target agent descends 30～50% than sodium channel blockers, IKr blocker proarrhythmia side effect incidence, and curative effect improves 30～40%, and the myocardial cell's of improvement electricity reconstruct effect is arranged.For example, the medicine that obtains according to the present invention has current potential time-histories under reach and hyperpolarization membrane potential feature, and these are obviously different with present known antiarrhythmic drug.According to inventor research, the medicine with such feature will effectively overcome antiarrhythmic drug unsatisfactory curative effect, proarrhythmia side effect, to problems such as patient's survival rate improvement effect are not good.
Embodiment:
Embodiment one: the described myocardial cell M of present embodiment
3Acceptor and IK
M3Purposes with screening treatment antiarrhythmic medicament.In extracellular fluid, add medicine to be screened, write down strength of current under the full cell pattern, the strength of current then ARR medicine of treatment for having curative effect that descends before than medication under 50mv voltage, aforementioned screening method can be used to screen inhibited medicine.
Embodiment two: present embodiment is for utilizing myocardial cell M
3Acceptor and IK
M3The method of screening the treatment antiarrhythmic medicament may further comprise the steps: a. separates single myocardial cell; B. set up full cell patch pincers detecting pattern; C. add M in the extracellular fluid
3Receptor stimulant; D. add other ionic channel retarding agent in the extracellular fluid; E. add the medicine of different solubility in the extracellular fluid, write down strength of current under the full cell pattern, strength of current then the ARR medicine of treatment that descends before than medication under 50mv voltage for having curative effect.
Detailed process is as follows:
A. separate single myocardial cell: separate single myocardial cell and obtain by the following method:
Laboratory animal is opened chest after with the anesthesia of 2% vetanarcol, take out heart rapidly, the aorta retrograde catheterization is connected on the Langendorff perfusion device, with the about 8ml.min of normal tyrode's solution
-1Perfusion is 5 minutes continuously, and the residual blood of flush away heart is used about 10 minutes of no calcium Zinciodati Comp solution perfusion then.After heart stopped to beat, it was unicellular to use the no calcium liquid perfusion isolating cardiac contain 0.05%II Collagen Type VI enzyme, when the heart deliquescing, begin the muscular tissue of coring when color and luster shoals, got 1 time every 2 minutes.More than operation is all at 37 ℃ of constant temperature and fill 95%O
2+ 5%CO
2Carry out under the condition, the cardiac muscular tissue that different time is taken off is put in the test tube that KB liquid is housed, and blows and beats gently with suction pipe, makes it to be dispersed into individual cells.Place 4 ℃ of refrigerators stablize 1 hour stand-by.Select shaft-like, anti-calcium, the clear agranular cell of band to experimentize.
Described laboratory animal refers to: cavy, rat, dog.
Described cardiac muscular tissue refers to: atrium or ventricular organization.
Described normal tyrode, preparation as follows: 136 NaCl, 5.4KCl, 1 MgCl2,0.33NaH
2PO
4, 5HEPES, 10 glucose and 1 CaCl
2Adjust pH to 7.4 with NaOH.
Described KB liquid, preparation (mM) as follows: 20 KCl, 10 KH
2PO
4, 25 glucose, 70 Potassium glutamates, 10 β hydroxybutyric acids, 20 taurines, 10 EGTA, 0.1% albumin and 40 N.F,USP MANNITOL are adjusted to 7.4 with KOH with pH.
B. set up full cell patch pincers detecting pattern: the full cell patch pincers of described foundation detecting pattern, undertaken by following scheme: used microelectrode draws with two-step approach and forms, and resistance was between 3～5 Ω after the microelectrode of making charged electrode solution.Ready microelectrode is connected on the patch clamp instrument probe the unicellular bath that places of separator well.With normal tyrode's solution with 3ml.min
-1Constant speed perfusion flushing cell surface after the formation high resistant sealing-in (10G Ω), is broken cytolemma with the pulsed suction, forms full cell pattern.Set protocol, stimulate with the voltage clamp pattern.Experimentation is by computer software control, and D/A is finished the generation of stimulus signal, the collection and the data analysis of feedback signal.
The described electrode solution that charges, by following scheme preparation (mM): GTP 0.1, Aspartic acid potassium 110, KCl 20, MgCl
21, Mg-ATP5, HEPES 10, and EGTA 10, and phosphocreatine 5 is adjusted pH to 7.3 with KOH.
The setting of described protocol, by following scheme through row: keep current potential-50mV; The stimulation voltage scope 2 seconds time length, returns to-30mV 1 second time length then from-40mV to+60mV; Stimulation voltage step 10mV, 5 seconds pitch times.
C. add M in the extracellular fluid
3Receptor stimulant: the described M that in extracellular fluid, adds
3Receptor stimulant is meant: 10mM choline (choline) or 10 μ M pilocarpines (pilocarpine).
D. add other ionic channel retarding agent in the extracellular fluid: described other ionic channel retarding agent that adds in extracellular fluid is meant: the how luxuriant and rich with fragrance Rider of 1mM, 20mM293B, 10mM U26452,200mM 4 aminopyridine and 200mM CdCL
2
E. add the medicine of different solubility in the extracellular fluid, write down strength of current under the full cell pattern, strength of current then the ARR medicine of treatment that descends before than medication under 50mv voltage for having curative effect.
It is provided by the invention that to be used to screen the antiarrhythmic drug action target be myocardial cell M
3Be the membranin that in irregular pulse, plays a crucial role, have mechanism of action characteristics such as hyperpolarization membrane potential, current potential time-histories under reach and anti-myocardial apoptosis.
Embodiment three: the screening process of present embodiment is: open chest after cavy is anaesthetized with 2% vetanarcol, take out heart rapidly, the aorta retrograde catheterization is connected on the Langendorff perfusion device, with the about 8ml.min of normal tyrode's solution
-1Perfusion is 5 minutes continuously, and the residual blood of flush away heart is used about 10 minutes of no calcium Zinciodati Comp solution perfusion then.After heart stopped to beat, it was unicellular to use the no calcium liquid perfusion isolating cardiac contain 0.05%II Collagen Type VI enzyme, when the heart deliquescing, begin the muscular tissue of coring when color and luster shoals, got 1 time every 2 minutes.More than operation is all at 37 ℃ of constant temperature and fill 95%O
2+ 5%CO
2Carry out under the condition, the heart muscle tissue that different time is taken off is put in the test tube that KB liquid is housed, and blows and beats gently with suction pipe, makes it to be dispersed into individual cells.Place 4 ℃ of refrigerators stablize 1 hour stand-by.Select shaft-like, anti-calcium, the clear agranular cell of band to experimentize.Normal tyrode is prepared as follows: 136 NaCl, 5.4KCl, 1 MgCl2,0.33 NaH
2PO
4, 5 HEPES, 10 glucose and 1 CaCl
2Adjust pH to 7.4 with NaOH.KB liquid is prepared (mM) as follows: 20 KCl, 10 KH
2PO
4, 25 glucose, 70 Potassium glutamates, 10 β hydroxybutyric acids, 20 taurines, 10 EGTA, 0.1% albumin and 40 N.F,USP MANNITOL are adjusted to 7.4 with KOH with pH.
Set up full cell patch pincers detecting pattern, undertaken by following scheme: used microelectrode draws with two-step approach and forms, and resistance was between 3～5 Ω after the microelectrode of making charged electrode solution.Electrode solution is prepared (mM): GTP 0.1 by following scheme, Aspartic acid potassium 110, and KCl 20, MgCl
21, Mg-ATP5, HEPES10, EGTA 10, and phosphocreatine 5 is adjusted pH to 7.3 with KOH.Ready microelectrode is connected on the patch clamp instrument probe the unicellular bath that places of separator well.With normal tyrode's solution with 3ml.min
-1Constant speed perfusion flushing cell surface after the formation high resistant sealing-in (10G Ω), is broken cytolemma with the pulsed suction, forms full cell pattern.Set protocol, stimulate with the voltage clamp pattern.Experimentation is by computer software control, and D/A is finished the generation of stimulus signal, the collection and the data analysis of feedback signal.The setting of described protocol, by following scheme through row: keep current potential-50mV; The stimulation voltage scope 2 seconds time length, returns to-30mV 1 second time length then from-40mV to+60mV; Stimulation voltage step 10mV, 5 seconds pitch times.
In extracellular fluid, add M
3Receptor stimulant 10mM choline (choline).In extracellular fluid, add other how luxuriant and rich with fragrance Rider of ionic channel retarding agent: 1mM, 20mM293B, 10mM U26452,200mM 4 aminopyridine and 200mM CdCL
2
Add the medicine of different solubility in the extracellular fluid, write down strength of current under the full cell pattern, strength of current then the ARR medicine of treatment that descends before than medication under 50mv voltage for having curative effect.
Embodiment four: the screening process of present embodiment is: open chest after cavy is anaesthetized with 2% vetanarcol, take out heart rapidly, the aorta retrograde catheterization is connected on the Langendorff perfusion device, with the about 8ml.min of normal tyrode's solution
-1Perfusion is 5 minutes continuously, and the residual blood of flush away heart is used about 10 minutes of no calcium Zinciodati Comp solution perfusion then.After heart stopped to beat, it was unicellular to use the no calcium liquid perfusion isolating cardiac contain 0.05%II Collagen Type VI enzyme, when the heart deliquescing, begin the muscular tissue of coring when color and luster shoals, got 1 time every 2 minutes.More than operation is all at 37 ℃ of constant temperature and fill 95%O
2+ 5%CO
2Carry out under the condition, the heart muscle tissue that different time is taken off is put in the test tube that KB liquid is housed, and blows and beats gently with suction pipe, makes it to be dispersed into individual cells.Place 4 ℃ of refrigerators stablize 1 hour stand-by.Select shaft-like, anti-calcium, the clear agranular cell of band to experimentize.Normal tyrode is prepared as follows: 136 NaCl, 5.4KCl, 1 MgCl2,0.33 NaH
2PO
4, 5 HEPES, 10 glucose and 1 CaCl
2Adjust pH to 7.4 with NaOH.KB liquid is prepared (mM) as follows: 20 KCl, 10 KH
2PO
4, 25 glucose, 70 Potassium glutamates, 10 β hydroxybutyric acids, 20 taurines, 10 EGTA, 0.1% albumin and 40 N.F,USP MANNITOL are adjusted to 7.4 with KOH with pH.
Set up full cell patch pincers detecting pattern, undertaken by following scheme: used microelectrode draws with two-step approach and forms, and resistance was between 3～5 Ω after the microelectrode of making charged electrode solution.Electrode solution is prepared (mM): GTP 0.1 by following scheme, Aspartic acid potassium 110, and KCl 20, MgCl
21, Mg-ATP5, HEPES10, EGTA 10, and phosphocreatine 5 is adjusted pH to 7.3 with KOH.Ready microelectrode is connected on the patch clamp instrument probe the unicellular bath that places of separator well.With normal tyrode's solution with 3ml.min
-1Constant speed perfusion flushing cell surface after the formation high resistant sealing-in (10G Ω), is broken cytolemma with the pulsed suction, forms full cell pattern.Set protocol, stimulate with the voltage clamp pattern.Experimentation is by computer software control, and D/A is finished the generation of stimulus signal, the collection and the data analysis of feedback signal.The setting of described protocol, by following scheme through row: keep current potential-50mV; The stimulation voltage scope 2 seconds time length, returns to-30mV 1 second time length then from-40mV to+60mV; Stimulation voltage step 10mV, 5 seconds pitch times.
In extracellular fluid, add M
3Receptor stimulant 10 μ M pilocarpines (pilocarpine).In extracellular fluid, add other how luxuriant and rich with fragrance Rider of ionic channel retarding agent: 1mM, 20mM293B, 10mM U26452,200mM 4 aminopyridine and 200mM CdCL
2
Add the medicine of different solubility in the extracellular fluid, write down strength of current under the full cell pattern, strength of current then the ARR medicine of treatment that descends before than medication under 50mv voltage for having curative effect.
Embodiment five: the screening process of present embodiment is: open chest after rat is anaesthetized with 2% vetanarcol, take out heart rapidly, the aorta retrograde catheterization is connected on the Langendorff perfusion device, with the about 8ml.min of normal tyrode's solution
-1Perfusion is 5 minutes continuously, and the residual blood of flush away heart is used about 10 minutes of no calcium Zinciodati Comp solution perfusion then.After heart stopped to beat, it was unicellular to use the no calcium liquid perfusion isolating cardiac contain 0.05%II Collagen Type VI enzyme, when the heart deliquescing, begin the muscular tissue of coring when color and luster shoals, got 1 time every 2 minutes.More than operation is all at 37 ℃ of constant temperature and fill 95%O
2+ 5%CO
2Carry out under the condition, the heart muscle tissue that different time is taken off is put in the test tube that KB liquid is housed, and blows and beats gently with suction pipe, makes it to be dispersed into individual cells.Place 4 ℃ of refrigerators stablize 1 hour stand-by.Select shaft-like, anti-calcium, the clear agranular cell of band to experimentize.Normal tyrode is prepared as follows: 136 NaCl, 5.4KCl, 1 MgCl2,0.33 NaH
2PO
4, 5 HEPES, 10 glucose and 1 CaCl
2Adjust pH to 7.4 with NaOH.KB liquid is prepared (mM) as follows: 20 KCl, 10 KH
2PO
4, 25 glucose, 70 Potassium glutamates, 10 β hydroxybutyric acids, 20 taurines, 10 EGTA, 0.1% albumin and 40 N.F,USP MANNITOL are adjusted to 7.4 with KOH with pH.
Set up full cell patch pincers detecting pattern, undertaken by following scheme: used microelectrode draws with two-step approach and forms, and resistance was between 3～5 Ω after the microelectrode of making charged electrode solution.Electrode solution is prepared (mM): GTP 0.1 by following scheme, Aspartic acid potassium 110, and KCl 20, MgCl
21, Mg-ATP5, HEPES10, EGTA 10, and phosphocreatine 5 is adjusted pH to 7.3 with KOH.Ready microelectrode is connected on the patch clamp instrument probe the unicellular bath that places of separator well.With normal tyrode's solution with 3ml.min
-1Constant speed perfusion flushing cell surface after the formation high resistant sealing-in (10G Ω), is broken cytolemma with the pulsed suction, forms full cell pattern.Set protocol, stimulate with the voltage clamp pattern.Experimentation is by computer software control, and D/A is finished the generation of stimulus signal, the collection and the data analysis of feedback signal.The setting of described protocol, by following scheme through row: keep current potential-50mV; The stimulation voltage scope 2 seconds time length, returns to-30mV 1 second time length then from-40mV to+60mV; Stimulation voltage step 10mV, 5 seconds pitch times.
In extracellular fluid, add M
3Receptor stimulant 10mM choline (choline).In extracellular fluid, add other how luxuriant and rich with fragrance Rider of ionic channel retarding agent: 1mM, 20mM293B, 10mM U26452,200mM 4 aminopyridine and 200mM CdCL
2
Add the medicine of different solubility in the extracellular fluid, write down strength of current under the full cell pattern, strength of current then the ARR medicine of treatment that descends before than medication under 50mv voltage for having curative effect.
Embodiment six: the screening process of present embodiment is: open chest after rat is anaesthetized with 2% vetanarcol, take out heart rapidly, the aorta retrograde catheterization is connected on the Langendorff perfusion device, with the about 8ml.min of normal tyrode's solution
-1Perfusion is 5 minutes continuously, and the residual blood of flush away heart is used about 10 minutes of no calcium Zinciodati Comp solution perfusion then.After heart stopped to beat, it was unicellular to use the no calcium liquid perfusion isolating cardiac contain 0.05%II Collagen Type VI enzyme, when the heart deliquescing, begin the muscular tissue of coring when color and luster shoals, got 1 time every 2 minutes.More than operation is all at 37 ℃ of constant temperature and fill 95%O
2+ 5%CO
2Carry out under the condition, the heart muscle tissue that different time is taken off is put in the test tube that KB liquid is housed, and blows and beats gently with suction pipe, makes it to be dispersed into individual cells.Place 4 ℃ of refrigerators stablize 1 hour stand-by.Select shaft-like, anti-calcium, the clear agranular cell of band to experimentize.Normal tyrode is prepared as follows: 136 NaCl, 5.4KCl, 1 MgCl2,0.33 NaH
2PO
4, 5 HEPES, 10 glucose and 1 CaCl
2Adjust pH to 7.4 with NaOH.KB liquid is prepared (mM) as follows: 20 KCl, 10 KH
2PO
4, 25 glucose, 70 Potassium glutamates, 10 β hydroxybutyric acids, 20 taurines, 10 EGTA, 0.1% albumin and 40 N.F,USP MANNITOL are adjusted to 7.4 with KOH with pH.
Set up full cell patch pincers detecting pattern, undertaken by following scheme: used microelectrode draws with two-step approach and forms, and resistance was between 3～5 Ω after the microelectrode of making charged electrode solution.Electrode solution is prepared (mM): GTP 0.1 by following scheme, Aspartic acid potassium 110, and KCl 20, MgCl
21, Mg-ATP5, HEPES10, EGTA 10, and phosphocreatine 5 is adjusted pH to 7.3 with KOH.Ready microelectrode is connected on the patch clamp instrument probe the unicellular bath that places of separator well.With normal tyrode's solution with 3ml.min
-1Constant speed perfusion flushing cell surface after the formation high resistant sealing-in (10G Ω), is broken cytolemma with the pulsed suction, forms full cell pattern.Set protocol, stimulate with the voltage clamp pattern.Experimentation is by computer software control, and D/A is finished the generation of stimulus signal, the collection and the data analysis of feedback signal.The setting of described protocol, by following scheme through row: keep current potential-50mV; The stimulation voltage scope 2 seconds time length, returns to-30mV 1 second time length then from-40mV to+60mV; Stimulation voltage step 10mV, 5 seconds pitch times.
In extracellular fluid, add M
3Receptor stimulant 10 μ M pilocarpines (pilocarpine).In extracellular fluid, add other how luxuriant and rich with fragrance Rider of ionic channel retarding agent: 1mM, 20mM293B, 10mM U26452,200mM 4 aminopyridine and 200mM CdCL
2
Add the medicine of different solubility in the extracellular fluid, write down strength of current under the full cell pattern, strength of current then the ARR medicine of treatment that descends before than medication under 50mv voltage for having curative effect.
Claims (3)
1. myocardial cell M
3Acceptor and I
KM3Purposes as screening treatment antiarrhythmic medicament is characterized in that myocardial cell M
3Acceptor and IK
M3Purposes with screening treatment antiarrhythmic medicament: add medicine to be screened in the extracellular fluid, write down strength of current under the full cell pattern, strength of current then the ARR medicine of treatment that descends before than medication under 50mv voltage for having curative effect.
2. the described myocardial cell M of claim 1
3Acceptor and I
KM3Purposes as screening treatment antiarrhythmic medicament is characterized in that described screening method is used to screen inhibited medicine.
3. myocardial cell M according to claim 1 and 2
3Acceptor and I
KM3Purposes as screening treatment antiarrhythmic medicament is characterized in that utilizing myocardial cell M
3Acceptor and I
KM3The method of screening the treatment antiarrhythmic medicament may further comprise the steps: a. separates single myocardial cell; B. set up full cell patch pincers detecting pattern; C. add M in the extracellular fluid
3Receptor stimulant; D. add other ionic channel retarding agent in the extracellular fluid; E. add the medicine of different solubility in the extracellular fluid, write down strength of current under the full cell pattern, strength of current then the ARR medicine of treatment that descends before than medication under 50mv voltage for having curative effect.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001029176A2 (en) * | 1999-10-15 | 2001-04-26 | Genaissance Pharmaceuticals, Inc. | Drug target isogenes: polymorphisms in the cholinergic receptor, muscarinic 3 gene |
| WO2002032924A2 (en) * | 2000-10-19 | 2002-04-25 | Genaissance Pharmaceuticals, Inc. | Haplotypes of the chrm5 gene |
| CN1515680A (en) * | 2003-01-06 | 2004-07-28 | 同济大学 | Application of ion channel inhibitor for curing arhythmia |
-
2004
- 2004-10-19 CN CNB2004100439458A patent/CN1306037C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001029176A2 (en) * | 1999-10-15 | 2001-04-26 | Genaissance Pharmaceuticals, Inc. | Drug target isogenes: polymorphisms in the cholinergic receptor, muscarinic 3 gene |
| WO2002032924A2 (en) * | 2000-10-19 | 2002-04-25 | Genaissance Pharmaceuticals, Inc. | Haplotypes of the chrm5 gene |
| CN1515680A (en) * | 2003-01-06 | 2004-07-28 | 同济大学 | Application of ion channel inhibitor for curing arhythmia |
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