CN104107431A - Improved tissue maintenance - Google Patents

Abstract

The invention provides an application of a composition which can be used for reducing damage to a tissue and organ during a reperfusion period, the composition comprises a potassium channel opener/agonist and/or an adenosine receptor agonist together with a local anaesthetic; wherein the damage is generated by reduction of activity induced by cardiac arrest, and the composition is suitable for being as a part for drug administration during a recovery stage in a surgical operation process.

Description

The tissue improving maintains

The application is to be on May 29th, 2006 applying date, and application number is 200680055410.X (corresponding international application no is PCT/AU2006/000717), and denomination of invention is the divisional application of the Chinese invention patent application of " tissue of improvement maintains ".

Technical field

The present invention relates to have no progeny protection and preserve tissue in homergy logistics body, as the heart for during stopping fighting.The invention still further relates to the method for the recovery for improving the hands postoperative patient after surgical operation.Especially, the present invention for minimize post-operative complication after operation on heart, particularly directly applies to heart instead of whole body especially after asystole.

Background technology

Annual global people up to a million select to open operation on heart, 1 to 3% patient will be dead in recovery room, 10% patient stays hospital because suffering from left ventricular dysfunction, and the patient up to 30% will be dead in 3 years by the high-risk patient of suffering from atrial arrhythmia and 24%.And the patient that creatine kinase (CK-MB) level after recent perspective study shows to perform the operation in blood raises a little has the risk of (3 to 5 years) M & M in significantly higher early stage (First Year) and late period.The iatrogenic ischemia reperfusion injury with during postoperative mortality rate and sickness rate and operation on heart of peri-operation period is relevant with unsuitable myocardial preservation.In addition, in department of pediatrics operation on heart, exceed that 50% baby has the myocardial damage of peri-operation period and with low cardiac output.In the past in 20 years, the type of an important part and cardioplegic solution in iatrogenic injury, form and send relevant.

In 2000, approximately 64% the cardiac operation of opening of carrying out was coronary artery bypass bypass surgery, the 24%th, and cardiac valve replacement or prosthesis and approximately 12% relate to the reparation of congenital heart defect.Approximately 1.2% is neonate/department of pediatrics.Major part is opened cardiac operation (exceeding 80%) need to use blood or lenticular asystole solution in cardiopulmonary bypass and selectivity asystole.At these intra-operatives, heart can stop to fight 3 hours, but sometimes reaches 4 to 6 hours.Amount of damage heart being caused in 3-4 hour is that the probability of heart restore funcitons reduces gradually, and higher with the probability compared with not having after stopping fighting 4 hours to recover.

At during surgery, conventionally make asystole or make it static with asystole fluid composition.Cardioplegic solution medicine is conventionally partly diluted and for example, mixes with carrier (4 parts of blood: the ratio of 1 part of crystal), or uses separately as crystal.The operation of small scale so-called " stop in a small amount rich liquid " (miniplegia) or " trace stops rich liquid " carry out under (microplegia), wherein a small amount of asystole solution/medicine and a large amount of blood (for example, 66 parts of blood: the ratio of 1 part of crystal) mix.Stop in a small amount rich liquid (Miniplegia) and be directly delivered to the tissue (for example heart) needing, and do not need to carry out sending of a large amount of whole bodies.Its objective is and make asystole and create " static depletion of blood region " for carrying out surgical operation, perioperative tissue injury (comprising in the time removing cardioplegic solution and cardiac resuccitation the potential substantive damage occurring in refilling process) is minimized.In nineteen fifty-five, doctor Melrose uses patient's self blood, as carrier, potassium citrate is sent into aorta so that asystole.In 1976, doctor Hearse described the administration of crystalloid cardioplegic solution.After several years, Buckberg and colleague propose to use patient's self blood as main carrier, because blood has oxygen carrying capability, swelling preferably and buffering property and endogenous antioxidant.Whole blood cardioplegic solution also with a large amount of blood and still less the potassium of titer carry out modification, be therefore called " stop in a small amount rich liquid ".It is to be created out by Menasche and colleague in the early 1990s that term " stops rich liquid " in a small amount.Stopping in a small amount rich liquid (or trace stops rich liquid) provides with the oxygen enrichment blood and the additive that stop rich microtitration liquid, so that asystole reduce ischemia reperfusion injury.

Ischemia injury depends on the persistent period of ischemic events, all still local in essence to a great extent.Ischemia is defined as between oxygen-supplying amount (coronary artery blood flow and oxygen carrying capacity) and oxygen demand (depending on contractility or the inotropic state of wall stress, heart rate and heart) and does not mate, and the seriousness of ischemia is to determine a key factor of secondary injury.Ischemic seriousness can compensate and even offset by increasing collateral flow amount.The basic premise of " stop in a small amount rich liquid " is to make ischemia and the damage that therefore causes minimizes.

According to definition, the heart aerobic metabolism maintaining during asystole needs oxygen-supplying amount to mate with oxygen demand.Therefore,, when oxygen demand in the induction of suitable cardioplegic solution and during maintaining asystole sharply reduces while exceeding 90%, for heart maintains aerobic metabolism, must meet a large amount of factors or pattern.These patterns can be summarized as follows: (i) oxygen must be satisfied the demand with enough existence, have now compellent evidence to show that hematocrit should be and at least equal 24%; (ii) oxygen must be sent to satisfy the demand with enough flow velocitys; (iii) oxygen should be sent to approach continuous mode as much as possible, and do not limit the surgical visual field, because it is (no matter the interval of described " safety " ischemia is in pilot model, in fact can not predict the metabolism of particular patient Myocardial and be converted to from aerobic mode degree and the reversibility of the tissue injury that (cut-off time mark) occurs before time point before anaerobic pattern and break time mark) by consuming in time; (iv) oxygen must be delivered to whole cardiac muscle as far as possible equably.When occur compactness narrow and and when coronary artery total blockage, there is now compellent a large amount of evidence to show: compared with independent direct motion administration, the method for (retrograde)/direct motion (antegrade) associating of driving in the wrong direction or more preferably drive in the wrong direction has been guaranteed being uniformly distributed of cardioplegic solution more effectively.

Although early stage perfusion again or recovery blood flow are the most effective myocardium modes of rescuing acute ischemia, the improper unexpected inflow of oxygen can cause necrosis, arrhythmia and death.The degree of " reperfusion injury " is associated with the cascade of inflammatory reaction (cascade), and described inflammatory reaction comprises the generation of cytokine, leukocyte, reactive oxygen species and free radical.Pouring into ischemic myocardial is that organizing of the final death of rescue is necessary again.But even the perfusion again after short-term ischemia is relevant to pathology variation, described pathology change list reveals the process itself causing between ischemic stage to be accelerated or pours into the rear new pathophysiology causing again to change.The degree of reperfusion injury and scope can be subject to the impact of inflammatory reaction in cardiac muscle.Ischemia-reperfusion promotes to discharge the proinflammatory medium of oxygen-derived free radicals, cytokine and other activation neutrophil cell and coronary blood endothelial tube.Described inflammatory process can cause endothelial function disturbance, blood capillary to be broken and blood flow is damaged, myocardial infarction and apoptosis.Pharmacology's anti-inflammatory therapeutics of targeting particular step has demonstrated and has reduced infarct size and myocardial damage.

From the beginning, low temperature has become a kind of fundamental of myocardial preservation.Its target is always reduced to the metabolism of ischemic interim minimum possible level, to maintain cardiac energy storage (adenosine three-phosphate and glycogen) and avoid tissue acidosis during ischemic episode.But many researcheres find to utilize 10 DEG C or 25 DEG C of not significant differences of the levels that cardiac muscle is recovered after crystalloid cardioplegic solution.Up to date; the main target of myocardial preservation is to protect myocyte's contractility to prevent pump failure; it comprise by reduce metabolism to low-level to preserve cellular energy, describedly low-levelly can continue to support important cytoactive such as ionic pump is to maintain interior environment.Not only there is very large interest to the heart during cardioplegic solution and general temperature in current technology, and cardioplegic solution is existed to very large interest to the impact of endothelium and blood capillary compartment.Therefore, the protection of endothelium may be protected no less important with myocyte.

Current technology still causes a large amount of patients to suffer postoperative atrial fibrillation.The Intensive Care Therapy that postoperative patient needs several days conventionally, and need to spend some times to return to clear-headed and mobility.These reflect that the reversible damage of some of them is to be caused by current operation, thereby need the technology of improving.

Summary of the invention

The reduction that the present invention relates to take in sail, i.e. the damage of the tissue, organ and the cell that cause of cell quiescent period, no matter its be caused by medical intervention or caused by other modes.Typically, it has the akinete of having a mind to for surgical operation object purposes, such as induce asystole for operation on heart.The present invention who describes is in this manual mainly the important business needs that maintain for improving tissue about the heart as organ.Medical science gets involved and comprises asystole, i.e. the asystole of medical science induction.But, the invention is not restricted to heart tissue, be suitable for equally and can be used for other organ, comprise neuronal tissue and cell, nephridial tissue, lung tissue, muscular tissue etc.Organ during the present invention also can be used for protecting the activity of non-medical induction to reduce under for example wound, shock, heart attack, apoplexy and similar incidents.

Term " organ " is in this article with its implication use the most widely, and any part that it refers to the health of exercising specific function, comprises tissue and cell or its part, for example cell line or organelle specimen.Other example comprises that causing circulatory is such as heart, respiratory apparatus are such as lung, urinary organs are such as kidney or bladder, digestive organs are such as stomach, liver, pancreas or spleen, genitals are such as scrotum, testis, ovary or uterus, nervous organ are such as brain, sexual cell are such as sperm or ovum and somatic cell are such as Skin Cell, heart cell are myocyte, neurocyte, brain cell or nephrocyte.

Term " tissue " is in this article with its implication use the most widely, and any part that it refers to the health of exercising specific function, comprises organ and cell or its part, for example cell line or organelle specimen.Other example comprises that conduit vascular is such as tremulous pulse or vein or causing circulatory are such as heart, respiratory apparatus are such as lung, urinary organs are such as kidney or bladder, digestive organs are such as stomach, liver, pancreas or spleen, genitals are such as scrotum, testis, ovary or uterus, nervous organ are such as brain, sexual cell are such as sperm or ovum and somatic cell are such as Skin Cell, heart cell (being myocyte), neurocyte, brain cell or nephrocyte.

Be to be understood that and " comprise (comprises) " or its phraseological modification is equivalent to term and " comprises (includes) " as term used in the present specification and claims, be interpreted as not get rid of to have other element or feature.

In one embodiment, the method that the invention provides tissue during a kind of activity decreased (for example, in order to promote surgical procedures) that reduces medical science induction, organ or cell injury, therefore it has reduced risk or the sickness rate of postoperative complication (such as the postoperative atrial fibrillation of Cardiac surgical procedures).Described method comprises the administration that comprises the compositions of potassium channel openers/agonist and/or adenosine receptor agonist (for example adenosine) and local anesthetic (for example lignocaine) in the time of perfused organ again.Preferably, use potassium channel openers/agonist or adenosine receptor agonist.

In a preferred form, directly described compositions is administered to tissue, organ or cell.Compositions described in (instead of as injecting agent (bolus)) administration in another embodiment, substantially continuously.In this article, " substantially continuously " allow less administration to interrupt.

Therefore, in another embodiment, the invention provides the compositions for reducing tissue and/or organ injury, described compositions comprises potassium channel openers/agonist and/or adenosine receptor agonist and local anesthetic.Described compositions may further include other component of listing as follows.In certain embodiments, described potassium channel openers/agonist and adenosine receptor agonist are replaced as calcium channel agonist by other ratio of component.

The present invention also provides the compositions that comprises potassium channel openers/agonist and/or adenosine receptor agonist and local anesthetic for the preparation of reducing tissue and/or organ injury, especially for the purposes reducing in the medicine of atrial fibrillation after the postoperative operation of Cardiac surgical procedures.

In one embodiment, described organ is heart.In this embodiment, compositions described in the time administration of cross icarceration can approximately removed.Once also can be in the i.e. compositions described in administration during administration asystole fluid composition of the operation that occurs asystole.

Preferably, described compositions comprises magnesium cation.Magnesium sulfate is its suitable source.

Preferably, described compositions administration between 0 DEG C to 37 DEG C.In certain embodiments, the temperature between 4 DEG C to 15 DEG C is suitable.In other embodiment, before administration, described compositions is heated to 20 DEG C to 37 DEG C.

The present invention also provides a kind of method, wherein compositions and/or compositions described in administration during wherein as perfused organ during resting state described in administration in the time of perfused organ again.This is commonly referred to surgical procedures and " maintains " phase.

In another embodiment, at administration cardioplegic solution as compositions described in administration before " pretreatment " step.Described pretreatment can be subject to compositions described in directly administration and be administered to the organ as surgical operation target or whole body experimenter's impact.Observe described pretreatment and there is protective effect.

Therefore, in a preferred step of the present invention, in the pre-induction that stops winning, maintenance period with as the compositions of convalescent a part of administration type described above of surgical procedures.

In one embodiment, the invention provides the method for fibrillation after a kind of operation reducing after operation on heart, described method comprises that administration comprises the compositions of potassium channel openers/agonist and/or adenosine receptor agonist and local anesthetic.Preferably, compositions described in administration between the convalescent period of described operation.

The present invention also provides compositions (particularly in the preferred embodiment as described below) for the purposes of method as mentioned above.This purposes of described compositions provides many treatment application, include but not limited to cardiovascular diagnosis (comprising the non-invasive diagnostic of coronary arteriography, myocardial scintigraphy, the conduction of two atrioventricular node), be used for the treatment of heart attack, resuscitation therapy, open operation on heart, angioplasty and other treatment get involved before, during or the organ-tissue that uses afterwards or short-term and the long term storage of cell (comprising grafting vessel).

In one embodiment, described compositions comprises adenosine and lignocaine.Especially, described compositions can comprise that weight ratio is adenosine and the lignocaine of about 1:0.5 to 4, particularly 1:2.

Be not subject to the constraint of any theory or model of action; think that protection relates to from regulating the multilayer system of membrane excitability to a large amount of intracellular signaling pathway; described intracellular signaling pathway causes: (i) reduce ion imbalance; the particularly sodium ion of load and calcium ion in cell; (ii) conductivity of improving atrium and ventricle is to coordinate metabolism needs; it can relate to adjusting gap connecting communication; (iii) vasodilation coronarius, and (ii) weaken to damage struvite replying.

Pretreatment and ischemia and again between flush phase described in infusion compositions the continuous protection tissue injury with ischemia resisting is provided, comprise and prevent fatal arrhythmia.When such as during angioplasty, stentplacement also being obtained in blood vessel to the protection that prevents local damage and inflammation.

Local anesthetic is for generation of reversible anesthetic medicine at body region.Many local anesthetics are made up of the aromatic rings that is connected to substituted-amino by carbochain containing carbonyl moiety.Conventionally, be defined as two class local anesthetics according to it containing carbonyl linking.Esters reagent comprises cocaine, tetracaine, procaine and chloroprocaine, and amide-type comprises prilocaine, mepivacaine, bupivacaine, mexiletine and lignocaine.Under high concentration, many medicines for other object have local anesthesia character.These comprise opium kind analgesics, beta-adrenoceptor antagonists, anticonvulsant (lamotrigine and lifarizine) and hydryllin.Can be selected from these type or derivatives thereofs according to the local anesthetic component of compositions of the present invention, or be selected from the medicine that can be used for other object.Preferably, described component also has local anesthesia character.

A kind of suitable local anesthetic is lignocaine.In this manual, lignocaine and lignocaine use interchangeably.Lignocaine is preferred, because it may be starting anaesthetic effect in the following way: blocking-up sodium express passway, suppress metabolic function, reduce free intracellular Ca2+, prevent from discharging in cell enzyme, may protect endotheliocyte and the damage of protection myofilament.Under lower treatment concentration, lignocaine has little effect to atrial tissue conventionally, and therefore it is invalid for treatment atrial fibrillation, atrial flutter and supraventricular tachycardia.Lignocaine is also free radical scavenger, anti-arrhythmic, and it has the character of antiinflammatory and the character of anti-Gao Ning.Also it must be understood that, under non-narcotic treatment concentration, local anesthetic blocking voltage compliance sodium express passway as may not can complete in lignocaine, enters but lower channel activity and reduce sodium.As anti-arrhythmic, it is believed that lignocaine targeting normally continues across the little sodium current of 2 phases of action potential, has therefore shortened action potential and refractory stage.

Lignocaine is a kind of local anesthetic, it is believed that its blocking-up sodium express passway, and has arrhythmia character by reducing the interior size to sodium current.In this manual, term " lignocaine " and " lignocaine " use interchangeably.It is believed that and follow the shortening of action potential directly to reduce calcium via Ca ²⁺selector channel and Na ⁺/ Ca ²⁺exchange enters cell.Latest report also show again between flush phase lignocaine with remove free radical in heart such as hydroxyl is relevant with singlet oxygen.The example of other suitable sodium channel blockers comprises that venom is such as Fugu ocellatus toxin and medicine primaquine, QX, HNS-32 (CAS registers #186086-10-2), NS-7, κ-opioid receptor agonist U50 488, Robenidine, pilsicainide, phenytoin, tocainide, mexiletine, RS100642, riluzole, carbamazepine, flecainide, Propafenone, amiodarone, sotalol, imipramine and moracizine or any its derivant.Other suitable sodium channel blockers comprises: vinpocetine (vinpocetine); B-carboline (ambocarb, AMB) with beta-carboline derivatives, nootropics activity.

Compositions according to the present invention comprises potassium channel openers.Potassium channel openers is to act on potassium channel to open the reagent of this potassium channel by door control mechanism.This causes potassium to flow out through film along its electrochemical gradient, described electrochemical gradient normally from cell to extracellular.Therefore, potassium channel is mediator, hormone or the pharmaceutically-active target spot that regulates cell function.Be to be understood that potassium channel openers comprises the potassium channel activator that also stimulates activity of potassium channels to produce identical result.Also be to be understood that the compound that has polytype opening or regulate different potassium channels; For example, some passage is voltage dependent, and some rectifier potassium channel is to ATP consumption, adenosine and opiates sensitivity, other activated by fatty acid, other passage regulates (, the passage of the variation of sodium and calcium in response cell) by ion ratio as sodium and calcium.Recently, have been found that diplopore potassium channel, it is believed that the background passage that it works to participate in regulating resting membrane electric potential.

Potassium channel openers can be selected from: nicorandil, diazoxide, minoxidil, pinacidil, aprikalim, cromokulim and derivant U-89232, P-1075 (a kind of selectivity plasma membrane KATP channel opener), emakalim, YM-934, (+)-7,8-dihydro-6, also [2,3-f] benzo-2 of 6-dimethyl-7-hydroxyl-8-(2-oxygen-piperidino)-6H-pyrans, 1,3-oxadiazole (NIP-121), RO316930, RWJ29009, SDZPCO400, rimakalim, symakalim, YM099, 2-(7,8-dihydro-6,6-dimethyl-6H[1,4] oxazines also [2,3-f] [2,1,3] Ben Bing oxadiazole-8-yls) pyridine N-oxides, 9-(3-cyano-phenyl)-3,4,6,7,9,10-six hydrogen-1,8-(2H, 5H)-acridine diketone (ZM244085), [(9R)-9-(the fluoro-3-125 iodophenyl of 4-)-2,3,5,9-tetrahydrochysene-4H-pyrans is-one-1, [3,4-b] thieno [2,3-e] pyridine-8 (7H) also, 1-dioxide] ([125I] A-312110), (-)-N-(2-ethoxyl phenenyl)-N'-(1,2,3-trimethyl propyl group)-2-nitroethylene-1,1-diamidogen (Bay X9228), N-(4-benzoylphenyl)-3,3, the fluoro-2-hydroxy-2-methyl of 3-tri-propionic acid amide. (ZD6169), ZD6169 (KATP opener) and ZD0947 (KATP opener), WAY-133537 and new dihydropyridine potassium channel openers A-278637.In addition, potassium channel openers can be selected from BK-activator (also referred to as BK-opener or BK (Ca)-class potassium channel openers or large conductance calcium-activated potassium channel openers) such as benzimidazolone derivatives NS004 (5-Trifluoromethyl-1-(chloro-2-hydroxyphenyl of 5-)-1, 3-dihydro-2H-2-ketone benzimidaozole), NS1619 (1, 3-dihydro-1-[2-hydroxyl-5-(trifluoromethyl) phenyl]-5-(trifluoromethyl)-2H-2-ketone benzimidaozole), NS1608 (N-(3-(trifluoromethyl) phenyl)-N'-(2-hydroxyl-5-chlorphenyl) urea), BMS-204352, retigabine (being also gaba agonist).Also comprise intermediate (for example benzoxazole, chlorzoxazone and zoxazolamine) and small-conductance calcium-activated potassium channel openers.

In addition, potassium channel openers can be used as indirect calcium antagonist, that is, they shorten the heart action potential time by accelerating 3 phase repolarizations, and thereby shortens plateau and reduce in calcium and flow to into cell and work.It is believed that reducing stream in calcium relates to L-type calcium channel, but also may relate to other calcium channel.

The direct calcium antagonist of certain embodiments of the present invention utilization, its Main Function is to reduce in calcium to flow to into cell.These are selected from least five kinds of main Types calcium ion channel blockor in greater detail as follows.Be to be understood that these calcium antagonists are by flowing to into cell and have potassium channel openers, particularly some effect of the potassium channel openers of ATP-sensitivity in inhibition calcium.

As potassium channel openers or agonist, adenosine is particularly preferred.Adenosine can be opened potassium channel, hyperpolarization cell, suppresses metabolic function, may protect endotheliocyte, increases the pretreatment of tissue and prevent ischemia or damage.The effect complexity of adenosine, has the character of many wide spectrums as medicine.Shown that adenosine increases coronary flow, hyperpolarization cell membrane and shields during ischemia and reperfusion.Adenosine also plays " in early days " or " delay " pretreatment " triggering agent " or reagent effect, prevents heart ischemia damage.Can think that the part of Cardioprotective character for adenosine is one or more adenosine receptor hypotypes of activation (A1, A2a, A2b and A3).The Signal Transduction Pathways that the protective effect of most of adenosine is attributed to A1 and A3 receptor activation and associating thereof causes pretreatment, protection and safeguards cell integrity.Also known adenosine participates in by activating A1 receptor the effect (adrenolytic) that slows down sinuatrial node pacing frequency (negative chronotropic's property (chronotropy)), delay chamber (A-V) knot conduction of impulse (negativity becomes conduction (dromotropy)), reduces atrial systole (negativity contractility) and inhibition catecholamine.The negative chronotropic of the adenosine that described A1-stimulates, become conduction reduce effect, the activation inward rectification potassium current (I of adenylate cyclase activity with inotropic effect and medicine _k-Ado) suppress, suppress phospholipid turnover, activate ATP-sensitivity K passage, suppress catecholamine to L-type Ca ²⁺the effect of electric current is relevant with nitricoxide synthase in activation AV ganglion cell.A3 receptor also demonstrates the effect with direct Cardioprotective, and A2 receptor has effective diastole and the antiinflammatory action of response damage.Adenosine is also a kind of calcium antagonist, vasodilator, anti-arrhythmic, antiadrenergic drug, free radical scavenger indirectly, stops agent (arresting agent), anti-inflammatory agent (weakening the activation of neutrophil cell), analgesic, metabolism agent and possible nitric oxide donors.

Be to be understood that antiadrenergic drug is such as beta blocker for example esmolol, atenolol, metoprolol and Propranolol can be used for replacing potassium channel openers or flow to into cell to reduce in calcium with its combination.Preferably, described beta blocker is esmolol.Similarly, α (1) thereby-adrenoceptor-antagonist is such as prazosin can be used for replacing described potassium channel openers or flows to into cell in calcium and reduce calcium load to reduce with its combination.

In one aspect of the invention, provide a kind of in ischemia or pretreatment between flush phase again, protection and/or reduce the compositions of tissue injury, described compositions comprises sends effective dose:

Antiadrenergic drug; With

Local anesthetic.

According to this this one side of the present invention, a kind of compositions is also provided, it comprises antiadrenergic drug and the local anesthetic of effective dose.

Preferably, described antiadrenergic drug is beta blocker.Preferably, described beta blocker is esmolol.

Also known adenosine suppresses Sodium/Calcium ion exchange indirectly, and it is by the load of sodium and calcium in minimizing cell.The inhibitor that is to be understood that described Sodium/Calcium ion exchange will cause reducing the effect of flowing and expand adenosine in calcium.Na ⁺/ Ca ²⁺exchanger inhibitor can comprise benzamyl, KB-R7943 (2-[4-(4-nitro benzyloxy) phenyl] ethyl] isothiourea mesylate) or SEA0400 (2-[4-[(2,5-difluorophenyl) methoxyl group] phenoxy group]-5-phenetidine).

Because one of character of adenosine is to reduce stream and sodium influx in calcium to enter in heart and coronary blood solencyte, should be further understood that before treatment, during treatment and the compound that causes stream and sodium influx in calcium to reduce (or reducing the calcium oscillation in cell) after treatment can be used for replacing in minimizing calcium, flow to into the adenosine of cell or with its combination.Such compound can be selected from the calcium ion channel blockor that three classes are different: Isosorbide-5-Nitrae-dihydropyridines (for example nitrendipine), phenylalkylamine class (for example verapamil) and benzothiazepine (for example diltiazem, nifedipine).

Calcium ion channel blockor is also referred to as calcium antagonist or calcium blockers.They are conventionally clinically for reducing heart rate and contractility and loose blood vessel.They can be used for the discomfort for the treatment of hypertension, angina pectoris or being caused by ischemia and some arrhythmia, and they have many effects of beta blocker (referring to above-mentioned discussion).

The calcium ion channel blockor of known five kinds of main Types with number of chemical structure: 1. benzothiazepine; For example diltiazem, 2. dihydropyridines: for example nifedipine, nicardipine, nimodipine and many other, 3. phenylalkylamine class: for example verapamil, 4. ammonia diaryl base propylamine ether: for example bepridil, the 5. naphthane of benzimidazole-replacement; For example mibefradil.

Conventional calcium ion channel blockor is combined in a large amount of L-type calcium channels (" slow channel ") that exist in cardiac muscle and smooth muscle, and what these medicine it contributes to be interpreted as has selectively acting to cardiovascular system.Dissimilar L-type calcium ion channel blockor is in conjunction with the different parts of α 1-subunit, and α 1-subunit is main passage-formation subunit (also having α 2, β, γ, δ subunit).The L-type passage that has different subclass, it contributes to tissue selectivity.Recently, also studied and there is not homospecific new calcium ion channel blockor, for example bepridil, it is that one also has Na except having L-type calcium channel blocking-up activity ⁺and K ⁺the medicine of carrier frequency channel break activity.Another example is mibefradil, and it has, and the blocking-up of T-type calcium channel is active and the blocking-up of L-type calcium channel is active.

Three kinds of common calcium ion channel blockors are diltiazem (Cardizem), verapamil (Calan) and nifedipine (Procardia).Nifedipine and relevant dihydropyridines do not have significant direct effect to atrioventricular conduction system or sinuatrial node under standard dose, therefore, conduction or self-disciplining are not had to direct effect.And other calcium ion channel blockor has negative chronotropic effect/dromotropic action (pacemaker activity/conduction velocity).For example, verapamil (with the diltiazem of degree still less) reduces the speed that in AV conducting system and SA knot, recover slow channel, therefore plays the active and conduction of slowing down of direct inhibitions SA knot pacemaker.These two kinds of medicines are frequency-and voltage dependents, make them more effective in cell, can rapid depolarization.Due to the probability of AV blocking-up or severe inhibition ventricular function, verapamil can not combine with beta blocker.In addition, mibefradil has negative chronotropic effect and dromotropic action.Calcium channel blocker (particularly verapamil) is also effective especially in treatment unstable angina pectoris, if related in angiospastic situation in its mechanism.

Omega conotoxins MVIIA (SNX-111) is a kind of N-type calcium ion channel blockor, it is reported that it as analgesic is 100-1000 times of morphine effect, but can be not addicted.Study this conotoxin and treating the pain of intractable (intractible).SNX-482, the toxin of the venom of a kind of Aranea of next reversal meat, blocking-up R-type calcium channel.Described compound separation is from the venom of African tarantula rust red bar cloth (Hysterocrates gigas), and it is the first R-type calcium ion channel blockor of describing.Can think that described R-type calcium channel works in the natural communication network of health, and contribute to regulate brain function.Other calcium ion channel blockor from regnum animale comprises the Kurtoxin from South Africa Scorpio, from the SNX-482 of African tarantula, climb very much the Taicatoxin of (Taipan) Serpentis from Australia, from the Agatoxin of funnel-web spider (Funnel Web Spider), from the Atracotoxin of Blue Mountains funnel-web spider (Funnel Web Spider), from the conotoxin of the Limax of Hai Sheng, from the HWTX-I of Chinese catching bird spider, from the Grammotoxin SIA of South America rose-red hair Aranea (Rose Tarantula).The derivant that also comprises these toxin with calcium antagonistic effect that this is listed.

Directly the potassium channel openers (for example nicorandil, aprikalem) of ATP-sensitivity or indirectly the potassium channel openers (for example adenosine, opiates) of ATP-sensitivity be also calcium antagonist indirectly, reduce this interior flowing to into tissue.For the potassium channel openers of ATP-sensitivity that also plays calcium antagonist effect, a kind of mechanism of thinking is shortened the heart action potential time and thereby shortened plateau by accelerating 3 phase repolarizations.During plateau, the net inflow amount of calcium may with the discharge balance of the potassium by potassium channel.Increasing by 3 phase repolarizations can be by blocking or suppressing L-type calcium channel and suppress to flow to cell in calcium and prevent calcium (and sodium) overload in histiocyte.

Can applicable therapeutic alliance comprise cardioplegic solution, there is no the processing of large grumeleuse (clot-busters) ischemic syndrome, cardiac operation (extracorporeal circulation and non-extracorporeal circulation (on and off-pump)), control arrhythmia, coronary artery is got involved (balloon and support), pretreatment organ, tissue or cell form ischemic stress, long-term organ or cell are preserved, peri-operation period or postoperative pain control, anti-inflammatory treatment after peri-operation period or operation, peri-operation period or hands Anticoagulation strategy (strategies), resuscitation therapy, cardiovascular diagnosis and other relevant treatment get involved.A kind of potential purposes for anticoagulant strategy can be used for the treatment of venous thrombosis with to surgical operation such as the relevant similar conditions of blood vessel, buttocks, heart and common surgical procedures and complication.

Calcium ion channel blockor can be selected from nifedipine, nicardipine, nimodipine, nisoldipine, lercanidipine, safe happy Horizon (telodipine), angizem, your stream tall and erect (altiazem), bepridil, amlodipine, felodipine, isradipine and cavero and other racemic variant.In addition, be to be understood that but the interior stream of calcium can be subject to the inhibition of other calcium blockers, described other calcium blockers can be used for replacing adenosine or with its combination, it comprises the in a large number venom from Hai Sheng or terrestrial animal, such as the omega-conotoxin GVIA (from snail conus geographus) of selective exclusion N-type calcium channel or respectively selective exclusion R-and P/Q-type calcium channel belong to ω-agatoxin IHA and the IVA of funnel-web spider (funnel web spider Agelelnopsis aperta) from canopy spider.Thereby also exist reduce calcium and sodium influx and contribute to the voltage-gated calcium of mixed type of Cardioprotective and sodium channel blockers such as NS-7.

Be to be understood that calcium ion channel blockor can be used for replacing local anesthetic or with its combination.

Therefore, in another aspect of the present invention, provide a kind of for pretreatment, protection and/or reduce ischemia or the compositions to tissue injury between flush phase again, it comprises sends effective dose:

Calcium ion channel blockor; With

Potassium channel openers or adenosine receptor agonist.

According to this this one side of the present invention, a kind of compositions is also provided, described compositions comprises calcium ion channel blockor and the local anesthetic of effective dose.Preferably, described calcium ion channel blockor is nifedipine.

In another embodiment, further comprise a kind of other potassium channel openers according to compositions of the present invention.Preferably, described other potassium channel openers is diazoxide.It is believed that diazoxide can protect ion and volume-adjustment, oxidative phosphorylation and mitochondrial membrane integrity (seemingly concentration compliance).Diazoxide also stress provide cardioprotection by the mitochondrial oxidation reducing when reoxygenation.Also exist some evidences to show that the protective effect of potassium channel openers is relevant with the generation of reactive oxygen species in adjusting mitochondrion.

Compositions according to the present invention comprises adenosine receptor agonist.Be to be understood that described adenosine receptor agonist comprises the while directly and indirectly causes receptor activation or simulation to have the compound of the effect of the receptor of identical net effect to receptor acting.

Suitable adenosine receptor agonist can be selected from: N ⁶-UK 80882 (CPA), N-ethyl formyl ammonia adenosine (NECA), p-(2-carboxyethyl) phenethyl-amino-5'-N-ethyl formyl ammonia adenosine of 2-[(CGS-21680), 2-chlorine adenosine, N ⁶-[2-(3,5-demethoxylation phenyl)-2-(2-methoxyphenyl] ethyl adenosine, the chloro-N of 2- ⁶-UK 80882 (CCPA), N-(4-aminobenzyl)-9-[5-(methyl carbonyl)-β-D-RIBOSE base (robofuranosyl)]-adenine (AB-MECA), ([1S-[1a, 2b, 3b, 4a (S*)]]-4-[7-[2-(the chloro-2-thienyl of 3-)-1-methyl-propyl group] amino]-3H-imidazoles [4,5-b] pyridine radicals-3-yl] cyclopentane formamide (AMP579), N ⁶-(R)-propyloxy phenyl base adenosine (R-PLA), aminophenyl ethyl adenosine 9APNEA) and cyclohexyladenosine (CHA).Other comprise that complete adenosine a1 receptor agonists is such as N-[3-(R)-tetrahydrofuran base]-adenine nucleoside (CVT-510) or partial agonist be such as CVT-2759 and allosteric reinforcing agent (allosteric enhancer) are such as PD81723.Other agonist comprises N ⁶the allosteric reinforcing agent of-cyclopenta-2-(3-phenyl amino carbonyl triazenes-1-yl) adenosine (TCPA) and A1 adenosine receptor; described TCPA is a kind of strong selectivity agonist for mankind A1 receptor with high-affinity, and described allosteric reinforcing agent comprises 2-amino-3-naphthoyl thiophene.CCPA is a kind of particularly preferred adenosine receptor agonist.CCPA is a kind of A1 adenosine receptor agonist.

Therefore, in yet another aspect, the invention provides a kind ofly for pretreatment, protection and/or reduce ischemia or the compositions of tissue injury between flush phase again, it comprises effective dose:

Potassium channel openers or adenosine receptor agonist;

Local anesthetic; With

CCPA。

Opiates is also known or be called opiates agonist, be the medicine that a class suppresses opium (Gr opion, Semen Papaveris juice) or morphine-sample character, it is conventionally clinically as extremely strong analgesic of moderate, especially, for controlling peri-operation period and postoperative pain.That other pharmacotoxicological effect of opiates comprises is sleepy, respiration inhibition, emotion change and mental clouding, and can not make loss of consciousness.It is believed that opiates is also the part of " triggering agent " in hibernation process, hibernation is a kind of dormancy form that is characterized as homergy rate and the decline of regular nuclear temperature.Under this hibernation-like state, tissue will be preserved to resist other to reduce the damage causing by oxygen or metabolism energy supply preferably, and prevents ischemic reperfusion injury.There are three kinds of opioid peptides: enkephalin, endorphins and dynorphin.

Opiates plays agonist effect, interacts with stereospecificity and saturable binding site in heart, brain and other tissue.Three kinds of main opiate receptors are identified and have cloned, i.e. μ, κ, δ receptor.Therefore, all these three kinds of receptors all belong to G-G-protein linked receptor family (its type comprises adenosine and bradykinin receptor).Opiate receptor has further hypotype, and for example, described δ receptor has two hypotypes, δ-1 and δ-2.

The Cardiovascular of opiates is positioned maincenter (, the cardiovascular of hypothalamus and brain stem and respiratory center) and the periphery (, myocardial cell and directly or indirectly vasoactive system) in complete health.For example, shown that opiates participates in vasodilation.Opiates can comprise the effect of direct opioid receptors or the indirectly effect of the non-opioid receptors of dose dependent to some effects of heart and cardiovascular system, such as the ion channel blocking of observing such as arylacetamide with the opiates with antiarrhythmic effect.Also known heart can synthesize or generate the opioid peptides of three types, i.e. enkephalin, endorphins and dynorphin.But, only in ventricular muscle cell, identify δ and kappa opioid receptor.

Be not subject to the constraint of any model of action, opiates is considered to provide cardioprotection by limiting ischemic damage and reducing ARR sickness rate, and it is for offsetting high-caliber damage agent or the compound of Spontaneous release during ischemia.This is to mediate via the activation of the potassium channel of the ATP sensitivity in sarolemma and mitochondrial membrane, and participates in open potassium channel.Further, also think that the cardioprotection of opiates is via the activation mediation of the potassium channel of the ATP sensitivity in sarolemma and mitochondrial membrane.Therefore, it is believed that opiates can be used for replacing potassium channel openers or adenosine receptor agonist or with its combination because they also participate in indirectly opening potassium channel.

Be to be understood that the compound that described opiates comprises directly and indirectly opiate receptor worked.The effect of opiates also comprises the effect of the non-opioid receptors of indirect dose dependent, such as the ion channel blocking of observing with the opiates with antiarrhythmic effect.

Therefore, in another aspect of the present invention, provide a kind of for pretreatment, protection and/and/or reduce ischemia and/or the compositions to organ, tissue or cell injury between flush phase again, described compositions comprises sends effective dose:

Opiates; With

Local anesthetic.

According to this aspect of the invention, also provide a kind of compositions, described compositions comprises anti-opiates and the local anesthetic of effective dose.Preferably, described opiates is selected from enkephalin, endorphins and dynorphin.Preferably, described opiates is the enkephalin of targeting δ, κ and/or μ receptor.More preferably, described opiates is selected from δ-1-opioid receptor agonist and δ-2-opioid receptor agonist.D-Pen2,5 enkephalins (DPDPE) are a kind of particularly preferred δ-1-opioid receptor agonists.

In another embodiment of the invention, provide a kind of according to compositions of the present invention, pharmaceutical composition of the present invention further comprises the antioxidant of effective dose.Antioxidant is common enzyme or other organic substance that can suppress the destruction of tissue oxygen.Can be selected from one or more in following according to the antioxidant component of compositions of the present invention: allopurinol, carnosine, histidine, coenzyme Q10, n-acetyl-cysteine, superoxide dismutase (SOD), glutathion reductase (GR), glutathion peroxidase (GP) regulator and adjusting agent, catalase and other metalloenzyme, NADPH and AND (P) H oxidase inhibitor, glutathion, U-74006F, vitamin E, Trolox (vitamin E of soluble form), other tocopherol (γ and α, β, δ), tocotrienol, ascorbic acid, vitamin C, beta-carotene (vitamin A of plant form), selenium, gamma linoleic acid (GLA), alpha-lipoic acid, uric acid (urate), curcumin, bilirubin, proanthocyanidin, EGCG salt, phylloxanthin, lycopene, bioflavonoids, polyphenol, trolox (R), dimethyl sulfourea, smooth bohr (R) (tempol (R)), carotenoid, ubiquinone, melatonin, flavonoid, polyphenol, amino indole, probucol and nitecapone, 21-aminosteroid or Lazaroids, compounds containing thiol groups (Thiazolidine, Ebselen, and N-acetylcystein dithiolethiones).Other antioxidant comprises the ACE inhibitor (captopril, enalapril, lisinopril) of the arterial hypertension and the heart failure that are used for the treatment of the patient who suffers from myocardial infarction.ACE inhibitor has useful effect by Scavenger of ROS bunch to the cardiac muscle of reoxygenation.Also operable other antioxidant comprises β-thin base propiono glycine, O-phenanthrolene, dithiocarbamate, selegilize and deferoxamine (Desferal); deferoxamine, iron chelating agent; it is for tentative cerebral infarction models, and wherein it brings into play the anti-oxidation protection effect of certain level.Spin trapping reagents ratio is if 5'-5-dimethyl-1-ketopyrrolidine (pyrrolione)-N-oxide (DMPO) and (a-4-pyridine radicals-1-oxide)-N-tert-butyl nitrone (POBN) are also as antioxidant.Other antioxidant comprises: nitrone radical scavenger α-phenyl-uncle-normal-butyl nitrone (PBN) and derivant PBN (comprising two sulfur derivatives); N-2-mercapto-propionyl glycine (MPG), a kind of specific scavengers of OH free radical; Lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA); Alpha lipoic acid; Chondroitin sulfate; Cys; Oxipurinol and zinc.

Preferably, described antioxidant is allopurinol (1H-pyrazolo [3,4-α] pyrimidine-4-alcohol).Allopurinol is a kind of competitive inhibitor of the reactive oxygen species that generates xanthine oxidase.The antioxidant properties of allopurinol can help protect cardiac muscle and endothelial function by reducing oxidative stress, mitochondrial injury, apoptosis and cell death.In addition, the system inflammation that protease inhibitor has weakened in the patient with extracorporeal circulation and other patient who just carries out operation on heart is replied, and other patient that inflammatory response has been enhanced is as AIDS or the patient who treats chronic tendon injury.Some broad-spectrum protease inhibitor is such as aprotinin also can reduce surgical operation such as the needs of losing blood and transfusing blood in bypass operation of coronary artery.

In another embodiment of the invention, the invention provides a kind of compositions, described compositions further comprises the nhe inhibitor of effective dose.Described nhe inhibitor has reduced sodium and calcium enters cell.Described nhe inhibitor can be selected from one or more in following: amiloride, cariporide, eniporide, triamterene and EMD 84021, EMD 94309, EMD 96785 and HOE 642 and T-162559 (Na ⁺/ H ⁺the inhibitor of the isoform 1 of ion exchange).Preferably, described nhe inhibitor is amiloride.Amiloride suppresses sodium proton ion exchange (Na ⁺/ H ⁺ion exchange, is also abbreviated as NHE-1 conventionally) and reduce calcium enter in cell.

During ischemia, excessive cell proton (or hydrion) is via Na ⁺/ H ⁺ion exchange exchange sodium.

Therefore, it is a kind of for pretreatment, protection and/or reduce ischemia or the compositions to tissue injury between flush phase again that another aspect of the present invention provides, and described compositions comprises sends effective dose:

Na ⁺/ H ⁺exchanger inhibitor; With

Local anesthetic.

Preferably, described Na ⁺/ H ⁺exchanger inhibitor is amiloride.

In another embodiment of the invention, the invention provides a kind of compositions, described compositions further comprises the magnesium source of effective dose, its content can increase the amount of magnesium in the cell of described tissue; And/or calcium source, its content can increase the amount of calcium in the cell of described tissue.

Protection during ischemia and the reoxygenation of the raising of magnesium and the minimizing of calcium and organ is relevant.This effect can be thought to be reduced and caused by calcium load.Preferably, the concentration that exists of magnesium is between 0.5mM to 20mM, more preferably from about 2.5mM.Preferably, the concentration that exists of calcium is between 0.1mM to 2.5mM, more preferably from about 0.3mM.In yet another aspect, also provide a kind of according to compositions of the present invention, it enters one and comprises the magnesium that effective dose improves.

Also can comprise non-penetrating agent (impermeant) or for minimizing or reduce the absorption of water by the cell of tissue according to compositions of the present invention.For by tissue cell minimize or reduce water absorb compound typically be non-penetrating agent (impermeants) or receptor antagonist or agonist.Be to control the transfer of water for minimize or reduce the compound object that water absorbs by the cell of tissue, that is, and the transfer of water between extracellular environment and intracellular environment.Therefore, these compounds participate in controlling or regulating osmosis.A kind of result be for by tissue cell minimize or reduce water absorb compound reduced the cell expansion relevant to edema (Cell swelling), such as the edema that can occur during ischemia injury.

Non-penetrating agent according to the present invention can be selected from one or more in following: sucrose, pentastarch, hetastarch, Raffinose, mannitol, gluconate, lactobionate and colloid.Colloid comprises albumin, hetastarch, Polyethylene Glycol (PEG), Dextran 40 and Dextran 60.Those of penetrant that can select to be included in for other compound of permeability object the main Types that regnum animale finds, comprise polyalcohols and saccharide, other aminoacid and amino acid derivativges and methylated ammonium and sulfonium compound.

Cell expansion also can cause by inflammatory response, and it may recover at organ, very important during preservation and surgical graft.Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 is a kind of important proinflammatory neuropeptide, and known its can cause cellular edema, and therefore the antagonist of Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 can reduce cell expansion.In fact, shown that antagonist (specificity neurokinine-1) receptor (NK-1) of Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 reduces the hepatic injury of inflammation, that is, edema forms, neutrophil cell infiltrates, hepatocellular Apoptosis and necrosis.Two kinds of such NK-1 antagonisies comprise CP96,345 or [(2S, 3S)-cis-2-(benzhydryl)-N-((2-methoxyphenyl)-methyl)-1-azabicyclic (2.2.2)-Xin-3-amine (CP-96,345)] and L-733,060 or [(2S, 3S)-3-([3,5-bis-(trifluoromethyl) phenyl] methoxyl group)-2-Phenylpiperidine].R116301 or [(2R-is trans)-4-[1-[3; 5-bis-(trifluoromethyl) benzoyl]-2-(benzyl)-4-piperidyl]-N-(2; 6-3,5-dimethylphenyl)-1-acetamide (S)-hydroxyl succinic acid ester] be another kind of activity specific neurokinine-1 (NK (1)) receptor antagonist; it has Ya Na meter mole (subnanomolar) affinity (K (i): 0.45nM) for mankind NK (1) receptor, has for NK (2) and NK (3) receptor the selectivity that exceedes 200 times.The antagonist that also can reduce the neurokinin receptor 2 (NK-2) of cell expansion comprises SR48968, and NK-3 comprises SR142801 and SB-222200.Also show in the brain sheet separating, the transmembrane potential that uses ciclosporin A to block mitochondrial permeability changes and minimizing mitochondrial inner membrane current potential has reduced the cell expansion of ischemia induction.In addition, glutamic acid-receptor antagonist (AP5/CNQX) and reactive oxygen species scavenger (ascorbic acid, Trolox (R), dimethyl sulfourea, Tan Boer (tempol (R))) also demonstrate and reduced cell expansion.Therefore, also can be selected from any in these compounds for the compound minimizing or reduce the Cell uptake water of tissue.

Also be to be understood that following energy substrate also can serve as non-penetrating agent.Suitable energy matter can be selected from one or more in following: glucose and other saccharide, pyruvate, lactate, glutamate, Glu, glutamine, aspartic acid, arginine, Ectoin (ectoine), taurine, N-acetyl group-beta-lysine, alanine, proline and other aminoacid and amino acid derivativges, trehalose, floridoside, glycerol and other polyhydric alcohol, sorbitol, inositol, pinitol, insulin, alpha-ketoglutarate, malate, succinate, triglyceride and derivant, fatty acid and carnitine and derivant.

Preferably, described for by tissue cell minimize or reduce water absorb compound be sucrose.Sucrose reduces water as non-penetrating agent to be shifted.Non-penetrating agent reagents ratio is as too large in sucrose, lactobionate and Raffinose and can not enter cell, therefore be retained in in-house intercellular substance, produce penetration to prevent cell expansion, described cell expansion damaged tissue by different way, it is by the lay up period generation at tissue especially.

Preferably, be approximately 5 to 500mM for the concentration minimizing or reduce the compound of the Cell uptake water of described tissue.Typically, this is the effective dose of the Cell uptake water for reducing described tissue.More preferably, be approximately 20 to 100mM for minimize or reduce the concentration of compound that water absorbs by the cell of tissue.Even more preferably, be about 70mM for minimize or reduce the concentration of compound that water absorbs by the cell of tissue.

In another preferred embodiment of the present invention, the invention provides a kind of compositions, described compositions comprises effective dose:

Potassium channel openers and/or adenosine receptor agonist; With

Local anesthetic,

One or more that further comprise effective dose are selected from following component:

Diazoxide;

Opioid;

Antioxidant;

Antiadrenergic drug;

Nhe inhibitor;

Calcium ion channel blockor;

Magnesium source; With

Calcium source.

Compositions of the present invention especially for comprising the happy dirty intra-operative pretreatment of heart transplantation, stop, protecting and/or preserve heart.Other application be included in cardiovascular get involved before, during or reduce afterwards heart and injury, described cardiovascular is got involved can comprise heart attack, " heart beating " surgical operation, angioplasty or angiography.For example, described compositions administration can be suffered from or developed into the experimenter of heart attack, and make the medicine of blood clot-break (busting) such as the time of streptokinase is used in administration.In the time of described clot dissolution, the described compositions cardioprotection of existence prevents further damage, such as reperfusion injury.

The organ that described compositions lacks (starved of) normal blood flow, nutriment and/or oxygen different time sections in extreme is such as those parts of heart are effective especially as protective agent.For example, described compositions can be used for ischemia that treatment is pre-existing in or that induced by medical science intervention.

In a preferred embodiment, compositions according to the present invention is compositions cardioplegic solution and/or Cardioprotective.

According to another aspect of the present invention, provide the purposes of compositions according to the present invention in the medicine for the preparation of pretreatment, protection and/or preservation organ.

In a preferred embodiment of this one side of the present invention, preferably before use and/or between the operating period described compositions is exposed in oxygen source.Described oxygen source can be the oxygen mixture that wherein oxygen is main component.Described oxygen can with for example CO ₂mix.Preferably, described oxygen mixture is 95% O ₂with 5% CO ₂.It is believed that with the oxygenate of oxygen mixture and maintain mitochondrial oxidation, this contributes to preserve the endothelium of myocyte and tissue.

Be to be understood that the amount of active component in described compositions will depend on experimenter's kind, the type of organ that stop, protecting and/or preserve and the application of proposal.Need the human experimenter of asystole at happy dirty intra-operative, the concentration of adenosine is preferably about 0.001 to about 2mM, more preferably from about 0.01 to about 10mM, most preferably from about 0.05 about 5mM, the concentration of lignocaine is preferably about 0.001 to about 2mM, more preferably from about 0.01 to about 10mM, and most preferably from about 0.05 to about 5mM.

For pretreatment, stop, keeping, make recovery or perfused organ, the concentration that is suitable for the adenosine that the form of the described compositions of embathing organ has is that 0.001mmols/ rises to 0.10mmols/ liter for maintaining with convalescent period, is 0.10 to 10mmols/ liter for stopping heart or other organ.As mentioned above, the concentration of lignocaine is conventionally in similar level, but Invariant and covariant can change.Ratio is the adenosine of 1:3: lignocaine is suitable for minimum maintaining and convalescent period.Can send described compositions by one or more in a large amount of approach, described approach comprises in intravenous, tremulous pulse, intraperitoneal, coronary artery approach in (direct motion or drive in the wrong direction), epidural and brain.

They can be with independent crystal or the multiple diluent administration to form to 64:1 (64 parts of blood: 1 part of crystal diluent) or higher blood ratio with 1:1 (1 part of blood: 1 part of crystal diluent), 1:4 (1 part of blood: 4 parts of crystal diluents) such as blood with carrier.

Described compositions is sent with multiple flow velocity conventionally.In withholding period, suitable flow velocity is 100 to 1000ml/min, and preferably 200 to 300ml/min, more preferably from about 350ml/min.Maintaining and convalescent period, sending for driving in the wrong direction, suitable flow velocity is 100 to 1000ml/min, and preferably 200-300ml/min, sends for direct motion, and suitable flow velocity is 10 to 200ml/min, and preferably 50 to 100ml/min.Those skilled in the art can regulate as required concentration and flow velocity with the activating agent (adenosine, lignocaine etc.) of sending optimal dose to organ.

In another preferred embodiment, by described compositions and physiology's carrier or crystal such as Plasma lyte ^tMor Normosol ^tMmix.In a preferred embodiment, in the time that described operation comprises extracorporeal bypass, additive (cassette) box is joined in the reperfusion solutions that comprises the adenosine (counting 12mg taking 3mg/ml) of 4ml, the magnesium sulfate of 10ml (as about 5g), and in the time recovering, add the lignocaine of 25mg.

A kind of preferred pharmaceutically acceptable carrier be have pH be approximately 6 to approximately 9, preferably approximately 7, more preferably from about 7.4 and/or low concentration for example about 10mM, the buffer of 2 to about 8mM, most preferably from about 4 to about 6mM potassium more preferably from about at the most.Suitable buffer comprises Cray Bai-Heng Zelaite solution (Krebs-Henseleit), tyrode's solution, Fremes solution, Hart Man (Hartmanns) solution and woods Ge Shi lactic acid solution (Ringers-Lactate), and described Cray Bai-Heng Zelaite (Krebs-Henseleit) comprises KCl, the NaHCO of 25mM of NaCl, the 5.9mM of glucose, the 117mM of 10mM conventionally ₃, 1.2mM NaH ₂pO ₄, 1.12mM CaCl ₂(free Ca ²⁺=1.07mM) and the MgCl of 0.512mM ₂(free Mg ²⁺=0.5mM), No. 2 (St.Thomas No.2) solution of St. Thomas, described tyrode's solution comprises KCl, the CaCl of 1mM of NaCl, the 5.4mM of glucose, the 126mM of 10mM conventionally ₂, 1mM MgCl ₂, 0.33mM NaH ₂pO ₄with the HEPES of 10mM (N-[2-ethoxy] piperazine-N'-[2-ethane sulfonic acid], described Hart Man (Hartmanns) solution comprises KCl, the CaCl of 2mM of 129 NaCl, 5mM conventionally ₂lactate with 29mM.An advantage that uses low potassium is that it can make compositions of the present invention less to experimenter's damage, particularly to department of pediatrics experimenter such as neonate/baby.High potassium is relevant with the accumulation of calcium, the accumulation of calcium may with convalescent period between irregular heart beat, heart and injury and cell expansion relevant.During asystole, neonate/baby is more vulnerable to the damage of high potassium than adult.After the surgical operation for damaged, neonate/baby's heart may still not recover normal after many days, sometimes needed to strengthen treatment or life support.Use have low concentration such as for example at the most the carrier of the magnesium of about 2.5mM be also favourable, but be to be understood that if expected, can use for example magnesium of about 20mM at the most of higher concentration, substantially can not affect the activity of described compositions.

In a further preferred embodiment, the invention provides a kind of medicine or veterinary compositions, described compositions comprises adenosine, lignocaine and pharmaceutically acceptable carrier, described pharmaceutically acceptable carrier comprises the potassium that is less than about 10mM.

Described compositions also can advantageously exist with the form of test kit, wherein preserves respectively active component for separately, continuously or administration simultaneously.

Be to be understood that compositions of the present invention also can comprise known medicine and/or be used in combination with it, this depends on proposed application.For example, the medicine that the adenosine in anti-Hemostatic Oral Liquid decomposes is substantially such as nucleoside transport inhibitor, and for example dipyridamole can be used as the additive in compositions of the present invention.The half-life of adenosine in blood is approximately 10 seconds, therefore, substantially prevents that the existence of the medicine of its decomposition from maximizing the effect that makes the present composition.Dipyridamole can advantageously add wherein to the concentration of about 10mM with about 0.1nM, and it has important benefit to Cardioprotective.The effect that dipyridamole can be increased vasodilative adenosine transport and supplemented adenosine by inhibition.When described in cyclical administration when compositions, this point is by particular importance.

The present invention also provides a kind of medicine or veterinary compositions, and it comprises active component and pharmacy or veterinary's acceptable carrier, diluent, auxiliary agent and/or excipient.

In theory, said composition and 100% a small amount of stop using together with rich liquid (miniplegia) (or trace stops rich liquid (microplegia)).Preferably, method for filling used should be followed the strict scheme that hot and cold cardioplegic solution is sent, and need not observe traditional myocardial preservation instruction (, you can use you often to think the volume of use) about time and volume constraint.

Astoundingly, the other benefit of the method has been improved the generation of urinating after corrective surgery, and without using diuretic.Conventionally, diuretic is added in the patient's who supports by heart-lung machine blood, but the needs of patients of finding to adopt the inventive method still less or do not need diuretic.Similarly, observe the method for the present invention of using, patient's lucidity is recovered quickly after operation.The natural law that needs severe disease monitoring still less-this can be by the result confirmation of following embodiment.

Detailed description of the invention

Just to setting forth the present invention by embodiment, carry out following EXPERIMENTAL DESIGN.This EXPERIMENTAL DESIGN utilizes a small amount of as above to stop rich liquid, the blood that wherein uses patient self oxygenate by micro-direct titration to heart." set (setting) " mentioned is the measuring of amount of substance that will directly be delivered to organ at pump on such as syringe pump, and described organ is heart in this embodiment.

Be prepared as follows two boxes.

(1) stop box (Arrest Cassette):

The undiluted potassium of 1.40ml, has 80mEq-and is equivalent to 2mEq/ml;

2. high setting: 25mEq's/ liter;

3. low setting: 10mEq's/ liter;

Potassium in above-mentioned project 1 is main asystole reagent.High potassium is well-known and the most the most frequently used cardioplegic solution, although it has known shortcoming and harmful side effect.A kind of alternative cardioplegic solution is disclosed in WO 00/56145 (GP Dobson), and it comprises potassium channel openers/agonist and/or adenosine receptor agonist (for example adenosine) and local anesthetic (for example lignocaine) in mM amount.Content in this description is all incorporated herein by reference.Although there is no example at this, the high potassium cardioplegia liquid of above-mentioned project 1 can replace with such cardioplegic solution.

(2) additive cartridge:

The adenosine of 1.4ml, contains 12mg-and is equivalent to 3mg/ml;

Magnesium sulfate=5g (or MgSO of a bottle of 2.10ml ₄, equal 5g);

Any crystal primer of 3.30ml-, can use (for example L/R, Plasma lyte in pump ^tM, Normosol ^tM);

4. the cumulative volume of additive cartridge: 44ml;

5. additive is set: 10ml/ liter;

This box is suitable for supporting the machine of 50ml box.

Lignocaine is joined in box as described below to send the result of improvement.Lignocaine is with 0.1 to 10 times of adenosine concentration, and preferably the concentration of 0.5 to 2 times adds.

Following data are from wherein lignocaine not being added in this box until remove the convalescent test before of cross icarceration at once.But, in another embodiment of the invention, lignocaine is added in this box for its first purposes, so that maintaining or the combination of administration resting stage adenosine and lignocaine in operation.Find that this has further improved the prospect of heart recovery and/or has reduced postoperative complication.

As follows for the method for compositions described in administration in this embodiment, overall goals be set up aerobicly stop, Ischemic stops winning.

1. in the time of heparinization, fill described ice bank to top with ice.Bank does not need to recharge, unless the time of x-icarceration exceedes 3 hours.Send temperature and will be approximately 12 DEG C.During approaching last 1/3rd x-icarceration, some metabolism of oxygen enrichment blood will be there will be.

2. temperature setting is set to for thermal induction: (37 DEG C) of temperature.

3. set for the height that stops winning: the potassemia that 25mEq/ rises stops box induction and stops fast.

4. for the setting of additive: 10ml/ liter before cross icarceration.

In the time of application cross icarceration:

1. for direct motion, increase fast flow to 500ml, be then retracted into immediately 320 to 350ml/min, to guarantee aortic valve closure.

2. give the warm direct motion of 700ml.Once obtain staticly, give 300ml many, be then switched to low K ⁺set (, 10ml/ liter).

3. give driving in the wrong direction of 700ml heat.

4. switch water temperature to cold.Give cold driving in the wrong direction the long as far as possible time.Reduce by rule of thumb and stop the rich long period of setting, flow and continue.

5. reduce additive and be set to 2ml/ liter, this appears in most of heart preparations.

6. if you are carrying out CABG, first carry out at far-end: after first set grafting, via many conduits, graft is hooked to pump with hook in system.Then, increase flow velocity to the pressure that obtains 150Torr very slowly, record flow, it is Useful Information for surgeon.This will realize some things:

■ controls mechanicalness device to utilize the goldstandard of pressure versus flow ratio to measure the unobstructed of graft;

■ surgeon has the method that checks anastomotic position hemostasis; With

The ability of driving in the wrong direction and sending when if ■ direct motion is delivered to target site and wants.

If 7. this operation relates to the valve and the coronary artery that work on, first carry out coronary artery.The method provides its nutriment needing to ill heart, and valve works on simultaneously.

8. according to common SOP monitoring K ⁺, and regulate potassium concn to reach the level of expectation.

In the time approaching last 10 minutes of x-icarceration, specimen is carried out to warm injection.These comprise:

1. water is set: (37 DEG C) of temperature.

2. stop rich setting: 0-is to rinsing out K ⁺and other metabolite.

3. the lignocaine of 25mg is injected to additive bag (described in this embodiment, do not add before) to complete the compositions of the preventative anti-arrhythmic of targeted delivery-now, in common described additive bag, retain about 18-35ml, this point depends on the persistent period of described operation, and the lignocaine concentration that it provides is about 1mg/ml.

4. additive is set: 15 to 18-before removing cross icarceration, target is except empty described auxiliary bag.

Injection for warm: conventionally remove delead in x-icarceration and start 5 to 10 minutes.

1. start the heating of driving in the wrong direction.Potassium is zero.Additive is set as 15.Guarantee that retrograde pressure maintains top level (35 to 40Torr).

2. in the time that electrical activity starts, then continue to drive in the wrong direction one minute.

3. switch to direct motion 2 to 3 minutes (in the time surgical blurred vision can not being made).This will promote degasification to transplant, and right side heart is poured, and conventionally will obtain stable heart rate.

4. switch the persistent period that is back to the x-icarceration of driving in the wrong direction.

If 5. additive has been set, continue to remove through x-icarceration with pure homoiothermy.

Stop winning liquid technology with trace, the capacity that you give is larger, heart stop winning better as aerobic in its former state.In many examples, if suitably administration, oxygen supply/aerobic ratio is reversible.Administration exceedes 1 and is raised at the most 6 liters and reduces relevant with the maximum of fibrillation after operation.

The observed value early that shows the damage of low temperature to some cell function with the clinical effectiveness that Warm blood cardioplegia obtains may be larger than the relation of anticipation before.These comprise and having reduced:

1. membrane stability;

2. utilize the ability of glucose and fatty acid;

3. the mitochondrion of adenosine triphosphate hydrochlorate generates, and causes suppressing the function of cell membrane;

4. the activity of adenosine triphosphate enzyme system, causes having damaged cell volume regulation;

5. sarcoplasmic reticulum reduces in conjunction with the ability of calcium;

6. mitochondrion state is breathed the activity of (Mitochondrial state respiration) and citrate synthetase;

7. the control of internal pH;

8. sarcoplasmic reticulum is for the activity of calcium absorption;

When approaching cross icarceration and finishing, the induction of associating temperature provides better result with cold maintaining with warm injection.Temperature induction, has particularly added adenosine (a kind of very strong vasodilator, also has other function) to open all sides and has propped up, and the additive for stopping rich necessary conduit and arrival myocyte and endothelium is provided.There will be with cold induction narrow, and can not entirety cardioplegic solution is distributed to myocyte and endothelium.

At the normal intra-operative of cross icarceration, due to ice-out, coldly the metabolism that provides is provided absorbs and reduce, slowly increasing appears in temperature.Mean temperature will change to approximately 12 to 14 DEG C.Last warm injection is the most important aspect of myocardial preservation.With heart-shaped paired photograph that relax, abiotic, that need contraction and electricity to support, by making heart experience homoiothermy (32 to 37 DEG C) the long as far as possible time, can making a difference in the functional rehabilitation that recovers most of hearts.Also evidence suggests and make the heart that is subject to wound cold, that relax, Lungs from Non-Heart-Beating stand high fluidity homoiothermy, such as standing in the time removing cross icarceration, can make heart guarantee to prevent reperfusion injury.

Between in the past 30 years, surgeon and perfusionist have improved their operating technology, make them can " customize " the concrete needs that how to make their method meet every patient.The region that only substantially retains " cooky forming machine (cookie cutter) " method is by myocardial preservation; Substantially " same size is applicable to all ".Be not subject to the restriction of specific theory and model of action, it is believed that the method for this preferred embodiment is to not having the patient of excessive hemodilution more responsive, thereby produced the result of improving.

In a test, 2688 patients that use cardioplegic solution to carry out operation on heart are assessed, in 6 different hospitals, use technology variation in this delicate environment with evaluation that different surgeon is different with it.With the high potassium cardioplegia liquid of standard process all patients with induction asystole.In patient, 1279 are in the group that stands typical standard crystal cardioplegic solution experimental design (" standard ").(1409 trace that stand the ALM additive cartridge that uses identical high potassium cardioplegia liquid and temperature as above stop rich liquid experimental design, a kind ofly use minimum cardioplegic solution directly to give heart), the ALM additive cartridge of described temperature is containing with good grounds compositions of the present invention.The present invention does not have specificity or is not limited the cardioplegic solution of this form, but it forms the application of method of the present invention, discusses for evaluating and set forth effect of the present invention at this.

Use as mentioned above described additive cartridge, so that during restoration it comprises adenosine, lignocaine and magnesium (therefore, being labeled as " ALM ").Method of the present invention is called " ALM ", and it is only abridged easily for one.According to experimental design as above, administration ALM in the time removing cross icarceration.

Table 1 has been listed 2688 patients' feature, and table 2 has been listed the event of the different postoperative complication of measuring.

In table 2, the clinical effectiveness of the patient of qualification in table 1 is made to table.Third column represents each result, and in patient, ALM ratio is as the percent (, the second field is as the percent on a hurdle) of standard cardioplegic solution Proportion of patients.The result on all left hurdles is all negative findings, and therefore minimizing of they expected.

Can find out at above-mentioned EXPERIMENTAL DESIGN infectious-related complication and substantially reduce, particularly postoperative atrial fibrillation and the needs to contractility in operation.Especially, being reduced to of these negative findingses: the contractility in operation has reduced by 86%; Pace-making in operation has reduced by 64%; Blood transfusion in operation has reduced by 44%; The postoperative hospital stays reduced by 21% and postoperative atrial fibrillation reduced by 91%.

Be to be understood that in this manual the present invention open and that limit extend in two or more words or accompanying drawing, mention or the therefrom all interchangeable combination of apparent independent characteristic.All these different combinations have all formed multiple interchangeable aspect of the present invention.

Claims (14)

1. compositions is in the application for the preparation of reducing again the medicament to tissue or organ injury between flush phase, and described compositions comprises:

Potassium channel openers/agonist and/or adenosine receptor agonist, and

Local anesthetic;

Wherein, the activity decreased that described damage is induced by asystole causes, and described compositions is to be suitable as the mode administration of a convalescent part of surgical procedures.

2. application according to claim 1, wherein said adenosine receptor agonist is adenosine.

3. application according to claim 1 and 2, wherein said local anesthetic is lignocaine.

4. application according to claim 1, wherein said compositions further comprises magnesium cation.

5. application according to claim 1, wherein, described compositions further comprises the pharmaceutically acceptable carrier with the potassium that is less than 10mM.

6. application according to claim 1, wherein, before administration, is heated at least 32 DEG C by described compositions.

7. application according to claim 1, is characterized in that, compositions described in administration in the time of perfused organ during resting state.

8. application according to claim 1, is characterized in that, described organ is heart.

9. compositions is in an application for the medicament for the preparation of fibrillation after reducing after operation on heart the operation between flush phase again, and described compositions comprises:

Potassium channel openers/agonist and/or adenosine receptor agonist, and

Local anesthetic;

Wherein said compositions is to be suitable as the mode administration of a convalescent part of surgical procedures.

10. application according to claim 9, wherein said adenosine receptor agonist is adenosine.

11. according to the application described in claim 9 or 10, and wherein said local anesthetic is lignocaine.

12. application according to claim 9, is characterized in that, described compositions further comprises magnesium ion.

13. application according to claim 9, is characterized in that, described compositions further comprises the pharmaceutically acceptable carrier with the potassium that is less than 10mM.

14. application according to claim 9, wherein, before administration, are heated at least 32 DEG C by described compositions.