CN1878464A

CN1878464A - Therapeutic delivery of carbon monoxide

Info

Publication number: CN1878464A
Application number: CN 200480028949
Authority: CN
Inventors: R·A·莫特利尼; R·A·阿尔伯托
Original assignee: Universitaet Zuerich; Hemocorm Ltd
Current assignee: Universitaet Zuerich; Hemocorm Ltd
Priority date: 2003-08-04
Filing date: 2004-08-04
Publication date: 2006-12-13
Also published as: GB0318254D0; GB0409376D0; ZA200601854B

Abstract

Boranocarbonates are described for administration to a human or other mammal for delivery of carbon monoxide. The boranocarbonate is a compound or ion adapted to make CO available for physiological effect, and may be administered with a guanylate cyclase stimulant or stabilizer. The physiological effect may be stimulation of neurotransmission, vasodilation or smooth muscle relaxation.

Description

The therapeutic of carbon monoxide is carried

Invention field

The present invention relates to be used for pharmaceutical composition and the compound with other mammal therapeutic ground conveying carbon monoxide to the people.Another purposes of said composition and compound is to be used for the organ perfusion.

Background of invention

Mammalian cell is by composing type (HO-2) and induction type (HO-1) Heme oxygenase ^1,2The ferroheme endogenous degraded that causes of family, can often produce carbon monoxide (CO) gas.CO is described to the neural courier that supposes at first ³, be considered to the multipotency signaling molecule now, have necessary regulating and controlling effect in its various physiology in betiding cardiovascular, nerve and immune system and the pathophysiological processes.In fact, the CO that is produced in vascular wall by Heme oxygenase has vasodilation character, and shows the inhibition vessel retraction and suppress acute and CH by the stimulation of soluble guanylate cyclase ^4-10Endogenous CO presents in liver circulation regulates dextrorotation tone (sinusoidal tone) ¹¹, control vascular smooth muscle cell propagation ¹²And the rejection that suppresses heart transplant ¹³The biological action of the CO of heme oxygenase enzyme source observed pharmacological effect when this gas externally being applied in the external and body system is confirmed.It is reported that in 10 to 500p.p.m. concentration range CO gas can mediate effective anti-inflammatory effect ¹⁴, prevent endothelial cell apoptosis ¹⁵, suppress people's tracheae smooth muscle cell proliferation ¹⁶With promote at contain oxygen high with the protection ischemic injury of lungs ^17,18In view of of the automatic adjusting of heme oxygenase enzymatic pathway at the control cell ¹⁹With the pleiotropy that manifests owing to CO ²⁰The key effect of middle performance it is contemplated that this diatomic gas can be used as the treatment tool that is used for the treatment of vascular function obstacle and immune related pathologies.

In the present invention, the method that three kinds of different being used to detect the treatment potentiality of CO has been proposed: 1) direct administration CO gas ²⁰2) use is subjected to the catabolic pro-drug of liver enzyme (being carrene) to produce CO ²¹With 3) by means of special CO carrier transport and release CO ²²Some researchers concentrate on up-to-date strategy process with their work, it is reported that promptly certain transition metal carbonyl has the ability of release CO under appropriate condition and has the function that discharges molecule (CO-RMs) in biosystem as CO.Specifically, demonstrate CO-RMs induction of vascular diastole in the arterial tissue that separates, and prevent that by special mechanism coronary artery shrinks and acute hypertension in the body, described mechanism can be simulated by the activation of HO-1/CO approach ²³What is interesting is that the chemistry of transition metal is multi-functional to be allowed by with the bio-ligand of metal center coordination they being modified effectively, so that make molecular toxicity littler, water-soluble stronger, and the release of adjusting CO.It is reported three carbonyl chlorine (the glycine root closes) rutheniums (II) (this paper is called CORM-3) recently; new synthetic water-soluble form metal carbonyl; it discharges CO in the biological model in external, stripped and body, protection cardiac muscle cell and tissue are avoided ischemia-reperfusion injury and heart allosome rejection ^24,25The part of this research is open by International Patent Application WO 02/092075 (list of references 25).

With regard to CORM-3, chloride and glycine ligand are unsettled, and they are present in the replacement of the high affinity ligands (being glutathione) in cell or the blood plasma environment, show as can accelerate CO and dissociate from metal center ²⁷In the time of in being added to the solution that contains myoglobin (Mb), CO has accelerated from the release of CORM-3, because every mole compound discharges 1 mole of CO in 1-2 minute ²⁴Therefore, CORM-3 belongs to the compound category that very rapidly discharges CO (" fast releaser "), and it can be used for wherein CO ideally as several clinical practices of signal medium matter (being neurotransmission, acute hypertension, angina pectoris, ischemia-reperfusion); Yet, identify the compound (" slow releaser ") that discharges CO with the slow motion mechanics, will realize the design of medicine, this medicine more multipotency ground is used for some to need chronic disease of the continuing of CO, permanent effect (is arthritis, inflammation, cancer, organ preservation; CH; The prevention of septic shock, sacculus postangioplasty restenosis, post operative ileus) treatment.

An interesting example in the research of transition-metal-carbonyl thing can be by describing for radiating the specially designed carbonyl of imaging technique, and wherein said transition-metal-carbonyl thing is used for not relating to the medical applications of CO therapeutical uses.Nearest technetium (I) complex of describing [ ^99mTc (OH ₂) ₃-(CO) ₃] ⁺Cause numerous interest as being used for the radiopharmaceutic precursor of technetium-99m ²⁸Many biomolecule, for example, peptide, scFV and CNS receptors ligand carry out mark by this method with technetium, confirmed [ ^99mTc (OH ₂) ₃-(CO) ₃] ⁺Be used for the potentiality that radiopharmaceutical is used ²⁹At CO and BH4 as reductant ^-Exist down, this compound can be in a step from water-based [ ^99mTcO ₃] ^-Be prepared ³⁰Yet, depend on the gaseous state carbon monoxide [ ^99mTc (OH ₂) ₃-(CO) ₃] ⁺Open preparation, and be not suitable for the purposes of commercialization radiopharmaceutical " kit ".Nearest research reported the first viable commercial in the physiology medium [ ^99mTc (OH ₂) ₃-(CO) ₃] ⁺Preparation, it uses the carbonylation agent based on boron, borine potash (K ₂[H ₃BCO ₂]), it is simultaneously as CO source and reductant ³¹

The physiological effect that discloses or pointed out borine carbonate (boranocarbonate) in the prior art.EP-A-34238 and EP-A-181721 have described the antitumor and hyperlipidemia disease activity of amine-carboxyl borine (carboxborane).US-A-4312989 discloses use amine-borine and has come the inflammation-inhibiting process.US-A-5254706 discloses the phosphorous acid-borane compound that is used for antitumor, anti-inflammatory and hyperlipidemia disease activity.

W093/05795 has discussed the organoboron compound purposes of anti-osteoporosis effectively, has also pointed out anti-inflammatory, hyperlipidemia disease and antineoplastic activity.Disclosed compound mainly is other compound of amino-boranes, but has also tested Na ₂BH ₃COO.People's such as Hall " Metal BasedDrugs ", Vol.2, No.1,1995, the anti-inflammatory activity of acyclic amine-carboxyl borine in the rodent has been described.

Because possible the effect of boron self or be similar to natural a-amino acid because of amino-boranes, these files shown the interest in the boride,

Summary of the invention

As illustration, the inventor has been found that the borine carbonate compound can be used for CO is transported to the physiology target so that physiological effect is provided by following detailed experiments data.

Therefore the invention provides and deliver medicine to people or other mammals and be applicable to CO is provided borine carbonate compound or ion and at least a pharmaceutically acceptable carrier with the performance physiological effect to discharge the pharmaceutical composition of carbon monoxide, to comprise.

Borine carbonate inaccurately is described as the compound group of the carboxylate adduct of borine and borane derivative.Borine carbonate contain usually connect the boron atom-group of COO-or COOR form (wherein R is H or other group), so they are called as borine carboxylate or carboxyl borine, but seem preferred term borine carbonate.In the list of references 31 compound K has been described ₂(H ₃BCOO) with relevant K (H ₃BCOOH), compound K wherein ₂(H ₃BCOO) be used to produce the Tc carbonyl.

Therefore, borine carbonate generally has the molecular structure that comprises following part:

Said structure preferably has three hydrogen atom (BH that connect boron ₃-CO-), be beneficial to CO and discharge because it is believed that this.

Also can be preferably wherein carboxylate groups connected the structure of boron, i.e. COO ^-,-COOH-,-COOX, wherein X can be pharmaceutically acceptable any suitable esterified group.

Borine carbonate compound in the pharmaceutical composition preferably has Formula B H _x(COQ) _yZ _zAnion, wherein :-

X is 1,2 or 3

Y is 1,2 or 3

Z is 0,1 or 2

x+y+z＝4，

Each Q is O ^-, represent the carboxylate anion form, or OH, OR, NH ₂, NHR, NR ₂, SR or halogen, wherein should or each R be alkyl (alkyl of preferred 1 to 4 carbon atom), each Z is halogen, NH ₂, NHR ', NR ' ₂, SR ' or OR ', wherein should or each R ' be alkyl (alkyl of preferred 1 to 4 carbon atom).

Because this general formula is similar to borine (borano) anion BH ₄ ^-, so this structure anion normally.When one (COQ) with (COO ^-) when existing, it can be a dianion.If this structure is an anion, then need cation.Can use on any physiology suitable cation, particularly metal cation, alkali metal cation for example is as K ⁺Or Na ⁺, or alkaline earth metal cation, for example Ca ⁺⁺Or Mg ⁺⁺Perhaps can use non-metal cations, for example NR ₄ ⁺, wherein each R is H or alkyl (preferred 1 to 4 carbon atom), or PR ₄ ⁺, wherein R is alkyl (alkyl of preferred 1 to 4 carbon atom).Cation is selected to reach the desired solvability of compound.

Y preferably 1.X preferably 3.

In composition, borine carbonate is preferably soluble, and exists in solution with The suitable solvent, for example aqueous solvent.Other possible solvent is a compatible solvent on ethanol, DMSO, DMF and other physiology.

The borine carbonate that uses among the present invention there are differences providing on the ability of CO.CO discharges can depend on pH and temperature.Low pH causes more or discharges faster.Therefore the compound scope that is used to select to be used for the suitable rate of release of special applications is available.Slow release is put and can be expended the long-time of a few hours or a couple of days.Can provide and contain the solution that dissolves CO, it is discharged by borine carbonate.Perhaps, can by change condition (as pH or temperature) or with by another kind of material (as another kind of solvent, or water-based physiological fluid, for example blood or lymph) contact, or and even discharge the release that CO is caused in the site at physiology.

Usually, pharmaceutical composition of the present invention discharges CO, makes with the dissolving form it to be provided to the therapeutic target.Yet CO can be directly released into the acceptor molecule of non-solvent in many cases.

Obviously, can carry CO by one or more above-mentioned mode of action therapeutic ground according to pharmaceutical composition of the present invention.

The borine carbonate compound also can comprise the target part that is beneficial at appropriate site release CO.Target part generally can discharge CO so that impel in the site of needs with receptors bind on specific target cell surface.This target part can be can with the part of the adjusting part of the receptors bind that exists on the target cell surface, perhaps can derive from another molecule, for example directly at the antibody of special receptor, this molecule is connected on the complex by suitable connector.

Pharmaceutical composition of the present invention usually contains pharmaceutically acceptable excipient, carrier, buffer, stabilizing agent or well known to a person skilled in the art other material.These materials should be nontoxic, and effectiveness that should the excessive interference active component.The accurate characteristic of carrier or other material depends on method of administration, in for example oral, intravenous, subcutaneous, nose, intramuscular, the peritonaeum, transdermal, through mucous membrane or the suppository approach.

Being used for pharmaceutical composition for oral administration can be tablet, capsule, pulvis or liquid form.Tablet can comprise solid carrier, for example gelatin or adjuvant or release polymer.Composition of liquid medicine comprises liquid-carrier usually, for example water, oil, animal oil or vegetable oil, mineral oil or artificial oil.Can comprise physiological saline, dextrose or other saccharide solution, or glycol for example ethylene glycol, propane diols or polyethylene glycol.Can also comprise the needed solvent of specific compound that contains in other solvent, particularly dissolved composition of pharmaceutically acceptable amount.

For intravenous injection, injection of skin or hypodermic injection, perhaps at ill site injection, active component generally is that solution is pyrogen-free and has suitable pH, isotonicity and stability with the outer acceptable solution form of enteron aisle.The technical staff that this area is relevant can prepare suitable solution well, for example use wait the carrier that oozes for example sodium chloride injection, ringer's inj, add the ringer's inj of lactic acid.Can comprise preservative, stabilizing agent, buffer, antioxidant and/or other additive as required.It also is known being used for the delivery system that Needleless injection uses simultaneously, therefore can prepare the composition that together uses with this system.

At the pharmaceutical composition that is used for discharging by any approach (including but not limited to mouth, nose, mucous membrane, vein, skin, subcutaneous and rectum), can be with active substance microcapsuleization in the polymerization ball, to such an extent as to be exposed to body fluid, and the CO that in time delays subsequently discharges.

Preferably carry out administration with prevention effective dose or treatment effective dose (depending on the circumstances, although prevention also can be regarded as treatment), this fully shows for individuality is favourable.The actual amount of administration, the speed of administration and time-histories depend on the character and the order of severity of being treated.Treatment prescription is for example determined dosage etc., within general doctor and other pharmacists' scope of offical duty, and generally will consider other factors known to the illness for the treatment of, single patient's site, medication and the doctor of situation, release.Above-mentioned technology and method example can be at Remington ' sPharmaceutical Sciences, 16th edition, and Osol, A. (ed) finds in 1980.

When preparing, must consider the toxicity of active component and/or solvent according to pharmaceutical composition of the present invention.Should consider the balance between medical effect and the toxicity.The dosage of general definite composition and formulation are so that the medical effect that provides surpasses any danger that causes owing to component toxicity.

In addition, provide a kind of CO has been imported mammiferous method, comprised the step of administration according to pharmaceutical composition of the present invention.Someone thinks that CO works by stimulating or activating guanylate cyclase at least in part.Someone thinks that CO has the especially effect of neurotransmitter and vasodilator.Therefore, provide the method that is used to stimulate the guanylate cyclase activity and discharges CO to mammal.The method that is used to excite nerve transmission or vasodilation and discharges CO to mammal also is provided.Yet the applicant does not wish to limit by theory, and does not get rid of the possibility that CO works by other mechanism.

Heme oxygenase 1 (HO-1) approach has been considered to represent the derivable system of defense of endogenous of maincenter, this system at stimulation comprise that UVA radiation, carcinogen, ischemia-reperfusion injury, endotoxic shock and some other produce the situation (32 that characterizes by oxygen radical, 19,2).The protective effect of HO-1 is owing to having produced effective anti-oxidants biliverdin and bilirubin and vasoactive gas CO.The expression of HO-1 connects (33) with the cardiac xenograft survival rate, also with the arteriosclerosis (34) of suppress transplanting and the myocardial dysfunction (35) after improving ischemic connect.In the paracmasis of rat acute inflammation, also directly relate to HO-1 (36).Other pathologic condition, for example the hemorrhagic shock of brain and liver and sepsis (37-39) is characterized in that having induced the HO-1 gene, and as if this gene play a part important in suppressing the vascular function obstacle that is caused by these pathophysiological states.As the consequence that HO-1 induces, the increase that CO produces influences the contractility of blood vessel significantly and reduces acute hypertension in the whole organism (10,9).Be exposed to animal in the surrounding air that contains low concentration CO or transfection HO-1 gene, caused preventing the injury of lungs that hyperoxia causes in the body, this is a kind of by weakening the mechanism (17,40) that neutrophil cell inflammation and lung apoptosis (cell death) mediate.Exogenous CO gas also has the ability (14) of the molecule I L-10 expression that suppresses former struvite cell factor in external and the body and regulate anti-inflammatory.Therefore, can be used for the treatment of all these situations, be used to regulate inflammatory conditions and extenuate other Pathological Physiology situation that comprises cancer according to administration CO medicine of the present invention.

Therefore, the invention provides a kind of method that imports CO to mammal, comprise the step of carrying out administration with pharmaceutical composition of the present invention, be used for the treatment of for example relevant acute and CH with lung, radiation damage, endotoxic shock, inflammation, the disease relevant with inflammation be asthma and rheumatoid arthritis for example, disease of intestine, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, organ damage behind the ischemic, miocardial infarction, angina pectoris, hemorrhagic shock, sepsis, penile erectile function obstacle and ARDS, and in sacculus angioplasty (ISR of treatment sacculus postangioplasty) and artery transplantation process.For example, the compound that it helps discharging CO in the sacculus angioplasty before the sacculus angioplasty and/or part afterwards discharge.Perhaps, support (stent) has the coating that comprises the compound that discharges CO.

Preparation be used for to the physiology target particularly mammal discharge CO so that the medicine of physiological action to be provided, the present invention also provides a kind of borine carbonate compound or ion as described herein, wherein physiological action for example is used to excite nerve and transmits or vasodilation, perhaps be used for the treatment of any hypertension, for example relevant acute and CH with lung, radiation damage, endotoxic shock, inflammation, the disease relevant with inflammation be asthma and rheumatoid arthritis for example, disease of intestine, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, sickle-cell anemia or sickle cell disease, organ damage behind the ischemic, miocardial infarction, angina pectoris, hemorrhagic shock, sepsis, penile erectile function obstacle and ARDS are in the process of for example sacculus angioplasty and artery transplantation.This this medicine be fit to by in oral, intravenous, subcutaneous, nose, inhalator, the muscle, in the peritonaeum, transdermal, through mucous membrane or the suppository approach carry out administration.

On the other hand, the invention provides the mammiferous method of treatment, comprise: by the transmission of exciting nerve of the CO as the physiology potent agent, vasodilation or smooth muscle relaxation, or treat any hypertension, radiation damage, endotoxic shock, inflammation, the disease relevant with inflammation, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, organ damage behind the ischemic, miocardial infarction, angina pectoris, hemorrhagic shock, sepsis, penile erectile function obstacle and ARDS, reangiostenosis, cirrhosis, cardiomegaly, heart failure and ulcerative colitis, or help the borine carbonate compound or the ion of CO performance physiological action by administration, in the sacculus angioplasty, treatment in the survival of artery transplantation or transplant organ.These are treatments relevant with the CO effect.

Preferably, methods of treatment is by the transmission of exciting nerve of the CO as the physiology potent agent, vasodilation or smooth muscle relaxation, or treat any acute or chronic systemic hypertension, radiation damage, endotoxic shock, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, organ damage behind the ischemic, miocardial infarction, angina pectoris, hemorrhagic shock, sepsis, the penile erectile function obstacle, reangiostenosis, cirrhosis, cardiomegaly, heart failure and ulcerative colitis, or in the sacculus angioplasty, treatment in the survival of artery transplantation or transplant organ.

More preferably, methods of treatment is by the CO as the physiology potent agent excite nerve transmission, vasodilation or smooth muscle relaxation, or treat damage that any acute or chronic systemic hypertension, hyperoxia cause, promote organ damage, angina pectoris, hemorrhagic shock, penile erectile function obstacle, cirrhosis, cardiomegaly, heart failure and ulcerative colitis behind cancer that apoptotic effect causes, graft-rejection, post operative ileus, the ischemic or the treatment in sacculus angioplasty or artery transplantation by CO.

Particularly, method can be the damage that causes of any hyperoxia of treatment, promote organ damage, angina pectoris, hemorrhagic shock, penile erectile function obstacle, cirrhosis, cardiomegaly, heart failure and ulcerative colitis behind cancer that apoptotic effect causes, graft-rejection, post operative ileus, the ischemic or the treatment in sacculus angioplasty or artery transplantation by CO.

By " smooth muscle relaxation " meaning is except by the treatment condition the vasodilation, for example chronic anal, musculus sphincter ani internus disease and anorectal disease.

Can use treatment more specifically that the present invention carries out and be the atherosclerotic group, the ischemic injury of lungs that suppress behind the artery transplantation, cardiac damage is induced in the prevention perfusion, also can realize the apoptotic effect of promotion (as in the cancer treatment) of CO.

The present invention also provides borine carbonate compound or the application of ion in treatment described here, and the ECP of the mammalian organs that can survive for example is for example in the storage and/or the transportation of the organ that is used for transfer operation.For this purpose, borine carbonate is the form of dissolving, is preferably dissolved in the aqueous solution.The organ that can survive can be any tissue that contains living cells, for example heart, kidney, liver, skin or muscle flap etc.

For example, can handle the organ of separation, as isolated organ or separate original position organ from blood supply.Organ can be, for example, and causing circulatory, respiratory apparatus, urinary organ, digestive organs, reproductive organs, nervous organ, muscle or flap or contain the man-made organ of living cells.Specifically, organ can be heart, lung, kidney or liver.Yet the bodily tissue that is subject to processing is not limited to and can is anyone or mammalian tissues, no matter be exsomatize or health in original position.We believe that also composition of the present invention can be used for the organ that exsomatizes or separate is discharged CO, so that reduce the ischemia injury of organ-tissue.

In the present invention, the borine carbonate described in the literary composition can be united use with guanylate cyclase stimulant or stabilizing agent, to carry CO to the physiology target, consequently provides the physiological action of enhancing.

In single composition, this pharmaceutical preparation can comprise borine carbonate and guanylate cyclase stimulant/stabilizing agent, or two components are prepared separately, to be used for simultaneously or in a sequence administration

Therefore the invention provides CO is imported mammiferous method as therapeutic agent, comprising: a) administration can provide the borine carbonate of the CO that is applicable to physiological action; And b) administration guanylate cyclase stimulant or stabilizing agent.

In aspect this, this method specifically is applied to treat organ damage behind acute or chronic systemic hypertension, pulmonary hypertension, graft-rejection, post operative ileus, arteriosclerosis, the ischemic, miocardial infarction, penile erectile function obstacle, reangiostenosis, cirrhosis, cardiomegaly, heart failure, chronic anal, musculus sphincter ani internus disease, anorectal disease and ulcerative colitis, perhaps is used for the treatment of sacculus angioplasty or artery transplantation.

Preferably, can first administration stabilizing agent/stimulant, administration borine carbonate subsequently, but this order can be put upside down.

Guanylate cyclase stabilizing agent/stimulant compound can be the compound that any stimulation guanylate cyclase produced or stablized guanylate cyclase, the activity form of guanylate cyclase specifically.Can use single composition maybe can use combination of compounds to be used for simultaneously or in a sequence administration, promptly various aspects comprise/use at least a guanylate cyclase stimulant/stabilizing agent.

Example comprise (3-(5 '-methylol-2 '-furyl)-1-benzyl-indazole) (YC-1), 4-pyrilamine-5-cyclopropyl-2-[1-[(2-fluorophenyl) methyl]-1H-pyrazolo [3,4-b] pyrimidin-3-yl] (BAY 41-2272), BAY 50-6038 (ortho position-PAL), BAY 51-9491 (a position PAL) and BAY 50-8364 (contraposition PAL).Accompanying drawing 9 shows the structure of ortho position, a position, contraposition PAL.Have been found that these compounds can be in conjunction with the avtive spot on the guanylate cyclase, and any compound that is similar in conjunction with this site can be used as guanylate cyclase stabilizing agent/stimulant.Also can use NO donor and 1-benzyl-3-(3 ¹-carbethoxyl group) phenyl-indazole, 1-benzyl-3-(3 ¹-methylol) phenyl-indazole, 1-benzyl-3-(5 ¹The lignocaine methyl)-furyl-indazole, 1-benzyl-3-(5 ¹-methoxy) furyl-indazole, 1-benzyl-3-(5 ¹-methylol) furyl-6-methyl-indazole, 1-benzyl-3-(5 ¹-methylol)-furyl-indazole-benzyl-3-(5 ¹-methylol)-furans-indazole, 1-benzyl-3-(5 ¹-methylol)-furyl-6-fluoro-indazole, 1-benzyl-3-(5 ¹-methylol)-furyl-6-methoxyl group-indazole, and 1-benzyl-3-(5 ¹-methylol)-and furyl-5,6-methylene dioxy indazole or its pharmaceutically acceptable salt.

For the patent application and the Patent right reason that relate to formerly, the applicant wishes to get rid of the purposes of following two kinds of compounds from the given protection in any aspect that claim of the present invention is claimed :-

I.K ₂(H ₃BCOO)

Wherein R, R '=H, alkyl, perfluoroalkyl.

Therefore randomly and temporarily implement this eliminating.

The medical treatment that the application relates to from start to finish comprises the people and treatment beast, and therefore the pharmaceutical composition that relates to comprises the composition that is used for people or beastly treatment.

Describe now and illustrate experimental data of the present invention.

In accompanying drawing subsequently, Fig. 1 to 8 is the experimental result pictures that show the following example 1 to 8.Fig. 9 is above-mentioned chemical formula.Figure 10 and Figure 11 are the figure as a result that shows the following example 9 and 10

Embodiment 1 to 8

Reagent

As previously mentioned, synthesize three carbonyl chlorine by Clark and partner and closed (the glycine root closes) ruthenium (II) ([Ru (CO) ₃Cl (glycinate)] or CORM-3) ²⁴As previously mentioned, synthesized CORM-A1 (Na by Alberto and partner ₂[H ₃BCO ₂], be designated as " CORM-A1 " here) ³¹Sodium borohydride (NaBH ₄) and all other reagent from SigmaChemicals (Poole, Dorset).

The preparation of passivity CORM-A1 and as the purposes of negative control

Preamble has been described the chemical property of borine carbonate in the aqueous solution ³¹This compound is metastable in the distilled water of alkaline pH.When pH when the more physiological condition (pH=7.4), compound begins to discharge CO, and the CO rate of release is very fast when acid pH.Based on this fact, we have generated the CORM-A1 (iCORM-A1) of passivity form by with compound and acid reaction.Specifically, the aqueous solution (100mM final concentration) that the concentrated hydrochloric acid (10M) of a small amount of five equilibrium (10 μ l) is added 1ml CORM-A1.Reaction causes the quick generation of gas (may be CO); Use stream of nitrogen gas that solution is carried out bubbling then, so that remove the residual CO gas of final dissolving.In having implemented CO is discharged the experiment that (being that Mb analyzes) and biological curative effect (being vasodilation) quantize, with this solution of five equilibrium negative control as CORM-A1.Because boron is the component of CORM-A1, and, also can utilize sodium borohydride (NaBH because during the release of the CORM-A1 from aqueous solution CO, can form boron hydride ₄) as the negative control in some experiments.

The detection that CO discharges

According to aforesaid method ²³, estimate CO and from CORM-A1, discharge by measuring spectrophotometric method that deoxidation myoglobin (Mb) is converted into carbon monooxide myoglobin (MbCO).By the absorbance (extinction coefficient=15.4M that measures 540nm in time at 37 ℃ ^-1Cm ^-1) growing amount of MbCO is quantized.Prepare myoglobin solution (the about 50 μ mol/L of final concentration) by protein being dissolved in 0.04M phosphate buffer (pH=7.4) temporarily.Add sodium dithionite (0.1%) so that the myoglobin of oxidation is converted into the myoglobin of reduction before each reading.According to different chemical and physical condition, the Mb in the time of also can using pH=5.5 or room temperature (room temperature) implements some experiments, so that detect the dynamics of CO from the release of CORM-A1.

The arterial ring that preparation separates: research vasodilation

From male Lewis rat, isolate the crosscut loop section of thorax artery, and by aforesaid method ¹⁰Under 37 ℃ its tension force low suspension at 2g is being equipped with in the organ bath of oxygen containing Krebs-Henseleit buffer solution.In with the pre-shrunk arterial ring of phyenlephrinium (3 μ M), estimate the stretching reaction of CORM-A1 (40,80 and 160 μ M).By inactive compound (iCORM-A1) or sodium borohydride (NaBH with equivalent ₄) add in the organ bath contrast ring is handled similarly.Also can be by relatively CORM-A1 and CORM-A3 are implemented experiment to the effect of vasodilation ground in time.

Embodiment 1. is by CO gas, and myoglobin (Mb) is to the conversion of carbon monooxide myoglobin

Be in the flesh red eggs hundred (Mb) of going back ortho states and be presented at the characteristic spectrum (referring to Fig. 1, dotted line) that the 555nm place has maximum absorption peak.When giving Mb solution (50 μ M) 1min, observe the quick conversion of carbon monooxide myoglobin (MbCO) with CO gas (1%) bubbling.As shown in Figure 1, MbCO show respectively 540 and the 576nm place have the characteristic spectrum (solid line) of two maximum absorption peaks.In the past after deliberation this method, with monitoring with detect the burst size of CO from CO-RMs ²³, and can be used for detecting how to influence the dynamics (referring to embodiment 4) that CO discharges under for example different pH of various conditions and the temperature.

Embodiment 2. is by CORM-A1, and myoglobin (Mb) is to the conversion of carbon monooxide myoglobin

CORM-A1 (60 μ M) is added in the Mb contain reduction (pH=7.4, the temperature=37 ℃) solution, cause producing gradually in time MbCO.As shown in Figure 2, behind incubation 210min, the standard spectrum of the Mb of reduction (filled squares) changes characteristic spectrum (empty del) into.The trace that contains asterisk is represented to carry out MbCO spectrum when saturated as Mb by the CO gas (positive control) described in materials and methods (Materials andMethods).

The dynamics that CO discharges from CORM-A1 under embodiment 3. room temperatures

After being added Mb solution, CORM-A1 produces the amount of MbCO, and can be by measuring extinction coefficient (ε=15.4M of known MbCO ^-1Cm ^-1) the absorbance at 540nm place quantize.The CORM-A1 adding with three kinds of variable concentrations at room temperature contains in the solution of Mb, and calculates the MbCO that produces in time.Use the nonlinear regression analysis of a phase exponential dependence (GraphPad Prism) to cause three curve (r ²＞0.99) best fit.As shown in Figure 3, the MbCO amount that produces from CORM-A1 increases according to the dynamics of determining in the mode that depends on concentration.For each figure, three kinds of concentration of the Ymax that is calculated (16.7 ± 1.2,33.1 ± 1.4 and 48.2 ± 2.5) and used CORM-A1 (be respectively 15.6,31.1 with 46.7 μ M) are very consistent.This shows and causes reaction that CO forms from CORM-A1 in time and trend is finished in aqueous solution, and shows that every mole compound discharges one mole of CO.At room temperature, CORM-A1 is 112 ± 3min from the mean half-life of matched curve.

Embodiment 4. temperature and pH are to the influence of CO from the CORM-A1 rate of release

Under different pH and temperature, detect the rate of release of CO from CORM-A1.Under three kinds of different conditions, CORM-A1 (60 μ M) is added in the Mb solution: 1) room temperature (room temperature) and pH=7.4; 2) 37 ℃ and pH=7.4; With 3) 37 ℃ and pH=5.5.As described in example 3 above, calculate the concentration of MbCO, and nonlinear regression analysis is used to obtain the best fit of three curves at different time points.As shown in Figure 4, CO is accelerated by the raising temperature with by reducing pH significantly from the rate of release of CORM-A1.Specifically, the half life period that can calculate CORM-A1 is 104min (triangle) at room temperature/pH=7.4, is 18.5min (rhombus) at 37 ℃/pH=7.4, and is 1.2min (square) at 37 ℃/pH=5.5.

Embodiment 5. compares the ability of CORM-A1 and passivity form (iCORM-A1) release CO thereof

Described in material and method (Materials and Methods) chapters and sections, when CORM-A1 was added acid solution, CO ran off apace.This process allows to produce inactive compound (iCORM-A1), and described compound can be used as the bioactive desirable negative control that is used to test these molecules.In order to confirm that iCORM-A1 can lose whole abilities that it discharges CO effectively, when the pH=7.4/ room temperature, compound (60 μ M) adding is contained in the solution of Mb (50 μ M), and calculate the MbCO that generates in time.As shown in Figure 5, iCORM-A1 (circle) can not produce any detectable MbCO, shows that compound is by complete passivation.Shown that CORM-A1 (square) to the influence that MbCO forms, is used for comparison.

Embodiment 6. compares the vasodilative ability of initiation of CORM-A1 and CORM-3

Shown CORM-3 ([Ru (CO) ₃Cl (glycinate)]) in the blood vessel that separates, promote diastole fast and significantly, and verified this effect is mediated by CO ²⁷From nearest research, also can learn the release of CO from CORM-3 to Mb or in biosystem take place very fast (about 5min) ^24,27, this meets the pharmacological effect of observing fast in the blood vessel that separates.With regard to CORM-A1, the release of CO (18.4min) is lower than the show value among the embodiment 5 under physiological pH.Therefore, the pharmacotoxicological effect that can expect CORM-A1 will reflect its biochemical property.In fact, as shown in Figure 6, compare with CORM-3 (80 μ M), CORM-A1 (80 μ M) produces very low influence to diastole.Specifically, join with the CORM-3 (solid line) in the pre-shrunk arterial ring of phyenlephrinium (Phe), in 4-5 minute, can promote 75% diastole, yet CORM-A1 (dotted line) causes progressive vasodilation, and vasodilation in 33 minutes is the highest (96%) after compound is added organ bath.

Embodiment 7.CORM-A1 is to the influence of vasodilative dependence concentration

Concentration (40,80 and 160 μ M) by the CORM-A1 that increases progressively is handled pre-shrunk arterial ring, and calculates vasodilative percentage in different time points.As shown in Figure 7, CORMA-1 causes significant diastole in time in the mode that relies on concentration.For example, from curve map as can be seen, following be 10 minutes after, the diastole percentage that is caused by the CORM-A1 of variable concentrations compared with the control: 40 μ M CORM-A1 are 21.0 ± 2.3%, 80 μ MCORM-A1 are 40.2 ± 3.4%, and 160 μ M CORM-A1 are 74.9 ± 1.8%.Tables of data is shown as the mean value ± S.E.M. of 6 independent experiments of each group.

The vasodilation character of embodiment 8.CO mediation CORM-A1

Pre-shrunk arterial ring is passed through 80 μ M CORM-A1, iCORM-A1 (inactive compound) or NaBH ₄Handle, this can be used as extra negative control (specifically referring to materials and methods).As shown in Figure 8, have only CORM-A1 to promote progressive and strong vasodilation, however iCORM-A1 and NaBH ₄All be fully invalid.These results clearly illustrate that CO is the immediate cause that causes the pharmacological effect observed from the release of CORM-A1.Tables of data is shown as the mean value ± S.E.M. of 6 independent experiments of each group.

Embodiment 9 and 10.

Before the experiment, compound is dissolved in prepares borine sodium carbonate (CORM-A1,100mM) liquid storage in the distilled water.From Sigma-Aldrich (Poole, Dorset) buy 3-(5 '-methylol-2 '-furyl)-1-benzyl-indazole (YC-1), and in dimethyl sulfoxide (DMSO) (DMSO), be prepared.Total data is expressed as mean value ± s.e.m..The difference between the evaluation analysis group is carried out in two tail t checks (Student ' s two-tailed t-test) by Student, utilization variance analysis (ANOVA) when relatively more than two kinds processing.O'clock think that the result has statistical significance in P＜0.05.

The arterial ring that preparation separates: research vasodilation

From male Lewis rat, separate the crosscut loop section of thorax artery, under 37 ℃ its tension force low suspension at 2g is being equipped with in the organ bath of oxygen containing Krebs-Henseleit buffer solution by preceding method (10).In with the pre-shrunk arterial ring of phyenlephrinium (1 μ mol/L), estimate the stretching reaction of CORM-A1 (20 μ M) when existing or lack YC-1 (1 μ M final concentration).30min before using the phyenlephrinium contraction adds YC-1 in the arterial ring that separates.

Zoopery: CORM-A1 and YC-1 are to the influence of blood pressure

Fentanyl citrate by intramuscular injection 1ml/kg is anaesthetized Lewis rat (280-350g).Surgery is implanted specially designed femoral artery and ductus venosus then, and uses several fluctuation tracer continuous monitoring mean arterial pressures (MAP) according to preceding method [23].Intravenous injection (i.v.) 50 μ mol kg ^-1Afterwards, estimate the influence of CORM-A1 in time to mean arterial pressure (MAP).By the 5min before adding the CORM-A1 pill to animals administer YC-1 (1.2 μ mol kg ^-1, i.v.), similarly test.Also can use YC-1 to implement control experiment separately.

Embodiment 9.CORM-A1 and YC-1 are to the influence of arteries diastole

With CORM-A1 pre-shrunk arterial ring is handled, and calculated the vasodilation percentage at different time points.As shown in figure 10, the CORMA-A1 of 20 μ M causes 13 ± 4.9% diastole behind the 20min; Be after with YC-1 (1 μ M) preliminary treatment blood vessel, to detect more obvious and more effective stretching reaction (61 ± 6.2%) enjoyably.Be noted that and in the contrast blood vessel pretreated and that shrink with phyenlephrinium, having only a spot of stretching reaction (being 2.8 ± 1.1% behind the 20min) with YC-1 in time separately.When 1 μ M and 10 μ M CORM-A1, also be very significant (being respectively 35 ± 9.8% and 51 ± 3.3%) with the stretching reaction of the pretreated blood vessel of YC-1.Tables of data is shown as the mean value ± s.e.m. of 6 independent experiments of each group. ^*P＜0.05 pair has only CORM-A1 or has only YC-1.

Embodiment 10.CORM-A1 and CORM-A3 are to the influence of mean arterial pressure

Femoral artery and ductus venosus surgery are implanted in the dopey Lewis rat, and monitored blood pressure continuously according to our preamble described [23].Figure 11 represents that CORM-A1 and YC-1 are to the influence of mean arterial pressure (MAP) in the body.With compound as pill, with the 50 μ mol/kg of CORM-A1 and the 1.2 μ mol kg of YC-1 ^-1Final concentration carry out intravenous injection.When two kinds of compounds carry out administering drug combinations, 10min administration YC-1 before the CORM-A1 injection.As shown in, CORM-A1 produces progressive and lasting reduction in time to MAP; For example, 60min after the CORM-A1 injection, MAP is reduced to 6.3 ± 1.5mmHg from initial baseline value.Injection YC-1 also can exert an influence to blood pressure separately; Yet the reduction of MAP is of short duration, reach 5.5 maximum ± 1.0mmHg after the 10min, and 50min gets back to foundation level after injection.What is interesting is that associating CORM-A1 and YC-1 can produce synergistic effect, cause quick and strong hypopiesia.In fact, MAP is reduced to 16.1 ± 5.6mmHg significantly after the 10min, and keeps this level for remaining experiment.Tables of data is shown as the mean value ± s.e.m. of 5 independent experiments of each group. ^*P＜0.05 pair baseline (10min); ^tP＜0.05 pair has only CORM-A1 or has only YC-1.

Therefore the invention provides as the water soluble compound that can have the CO carrier of chemical property optionally with based on what CO discharged new methods of treatment is provided.This method is better than sucking CO significantly, because it can avoid CO gas that the related problem of the systemic effects of oxygen transportation and release is taken place.In addition, it is very feasible having the design that CO discharges " soon " or " slowly " dynamic (dynamical) stable compound, this compound energy selectivity target organs wherein, and only act on the limited district of health.An application of water soluble compound purposes is the topical application Co that needs in condition.For example, in order to protect vascular tissue and to prevent reangiostenosis, the compound that can will supply with CO before sacculus angioplasty process is applied to blood vessel during the sacculus angioplasty.Perhaps, available special borine carbonate compound wraps quilt to intravascular stent, and this compound has to damaged blood vessels and slowly discharges CO and suppress the ability of smooth muscle cell proliferation.The compound of the dynamics temperature influence that CO discharges also can be used as the adjuvant of impregnating fluid by the use of exsomatizing, and this impregnating fluid is preserved organ before being used in usually and transplanting.Wide coverage the protective effect of HO-1 at the organ rejection, and the idea of treatment of organs rather than the acceptor clinical testing that will be very beneficial for transplanting.

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Claims

1. borine carbonate compound or ion are in the purposes of preparation in the medicine, this medicine with CO as the transmission that is used to excite nerve of physiology potent agent, vasodilation or smooth muscle relaxation, perhaps be used for the treatment of any hypertension, radiation damage, endotoxic shock, inflammation, the disease relevant with inflammation, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, organ damage behind the ischemic, miocardial infarction, angina pectoris, hemorrhagic shock, sepsis, the telotism dysfunction, ARDS, reangiostenosis, cirrhosis, cardiomegaly, heart failure and ulcerative colitis perhaps are used for the sacculus angioplasty, treatment in artery transplantation or the transplant organ survival.

2. purposes according to claim 1, wherein said medicine with CO as the transmission that is used to excite nerve of physiology potent agent, vasodilation or smooth muscle relaxation, perhaps be used for the treatment of any acute or chronic systemic hypertension, radiation damage, endotoxic shock, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, organ damage behind the ischemic, angina pectoris, hemorrhagic shock, sepsis, the telotism dysfunction, reangiostenosis, cirrhosis, cardiomegaly, heart failure and ulcerative colitis perhaps are used for the sacculus angioplasty, treatment in artery transplantation or the transplant organ survival.

3. purposes according to claim 1, wherein said medicine with CO as the transmission that is used to excite nerve of physiology potent agent, vasodilation or smooth muscle relaxation, perhaps be used for the treatment of any acute or chronic systemic hypertension, the damage that hyperoxia causes, promote the cancer that apoptotic effect causes by CO, graft-rejection, post operative ileus, organ damage behind the ischemic, angina pectoris, hemorrhagic shock, the penile erectile function obstacle, cirrhosis, cardiomegaly, heart failure and ulcerative colitis perhaps are used for the treatment of sacculus angioplasty or artery transplantation.

4. according to any described purposes in the claim 1,2 and 3, wherein said medicine be fit to by in oral, intravenous, subcutaneous, nose, inhalator, the muscle, in the peritonaeum, transdermal, carry out administration through approach mucous membrane or suppository.

5. according to any described purposes in the claim 1 to 4, wherein the molecular structure of borine carbonate compound or ion comprises following part

6. purposes according to claim 5, wherein borine carbonate compound or ion comprise BH ₃-CO-part.

7. according to claim 5 or 6 described purposes, wherein borine carbonate is Formula B H _x(COQ) _yZ _zCompound or anion,

Wherein :-

X is 1,2 or 3

Y is 1,2 or 3

Z is 0,1 or 2

x+y+z＝4，

8. purposes according to claim 7, wherein z is 0.

9. according to Claim 8 or 9 described purposes, wherein y is 1.

10. purposes according to claim 7, wherein x is 3.

11. according to any described purposes in the claim 7 to 10, wherein borine carbonate is the anion with Q of at least one O-or OR form, and described composition comprises at least one metal cation.

12. purposes according to claim 11 wherein is somebody's turn to do or each metal cation is alkali metal cation or alkaline earth metal cation.

13. purposes according to claim 12, wherein borine carbonate is Na ₂(H ₃BCO ₂).

14. according to any described purposes in the claim 1 to 13, wherein said medicine also comprises guanylate cyclase stimulant or stabilizing agent.

15. purposes according to claim 14, wherein guanylate cyclase stimulant or stabilizing agent be not with the molecule or the ion of borine carbonate compound or ions binding.

16. claim 14 or 15 described purposes, wherein guanylate cyclase stimulant or stabilizing agent are YC-1.

17. according to any described purposes in the claim 14 to 16, a kind of in the administration of administration and order when wherein said medicine is suitable for borine carbonate compound or ion and guanylate cyclase stimulant or stabilizing agent.

18. according to any described purposes in the claim 1 to 17, wherein borine carbonate compound or ion are different from:

I. K ₂(H ₃BCOO)

Wherein R, R '=H, alkyl, perfluoroalkyl.

19. mammiferous method of treatment, comprise by administration borine carbonate compound or the ion that provides CO to bring into play physiological action is provided, with CO as the transmission of exciting nerve of physiology potent agent, vasodilation or smooth muscle relaxation, perhaps treat any hypertension, radiation damage, endotoxic shock, inflammation, the disease relevant with inflammation, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, organ damage behind the ischemic, miocardial infarction, angina pectoris, hemorrhagic shock, sepsis, the telotism dysfunction, ARDS, reangiostenosis, cirrhosis, cardiomegaly, heart failure and ulcerative colitis are perhaps in the sacculus angioplasty, treatment in artery transplantation or the transplant organ survival.

20. method according to claim 19, comprise with CO as the transmission of exciting nerve of physiology potent agent, vasodilation or smooth muscle relaxation, perhaps treat any acute or chronic systemic hypertension, radiation damage, endotoxic shock, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, organ damage behind the ischemic, angina pectoris, hemorrhagic shock, sepsis, the telotism dysfunction, reangiostenosis, cirrhosis, cardiomegaly, heart failure and ulcerative colitis are perhaps in the sacculus angioplasty, treatment in artery transplantation or the transplant organ survival.

21. method according to claim 19, comprise with CO as physiology potent agent excite nerve transmission, vasodilation or smooth muscle relaxation, perhaps treat damage that any acute or chronic systemic hypertension, hyperoxia cause, promote organ damage, angina pectoris, hemorrhagic shock, penile erectile function obstacle, cirrhosis, cardiomegaly, heart failure and ulcerative colitis behind cancer that apoptotic effect causes, graft-rejection, post operative ileus, the ischemic, the perhaps treatment in sacculus angioplasty or the artery transplantation by CO.

22. according to any described method in the claim 19,20 or 21, comprising by in oral, intravenous, subcutaneous, nose, inhalator, the muscle, in the peritonaeum, transdermal, carry out administration through approach mucous membrane or suppository.

23. according to any described method in the claim 19 to 22, wherein the molecular structure of borine carbonate compound or ion comprises following part:

24. method according to claim 23, wherein borine carbonate compound or ion comprise BH ₃-CO-part.

25. according to claim 23 or 24 described methods, wherein borine carbonate is Formula B H _x(COQ) _yZ _zCompound or anion, wherein :-

X is 1,2 or 3

Y is 1,2 or 3

Z is 0,1 or 2

x+y+z＝4，

26. method according to claim 25, wherein z is 0.

27. according to claim 25 or 26 described methods, wherein y is 1.

28. method according to claim 25, wherein x is 3.

29. according to any described method in the claim 25 to 28, wherein borine carbonate is the anion with Q of at least one O-or OR form, and described composition comprises at least one metal cation.

30. method according to claim 29 wherein is somebody's turn to do or each metal cation is alkali metal cation or alkaline earth metal cation.

31. method according to claim 29, wherein borine carbonate is Na ₂(H ₃BCO ₂).

32. according to any described method in the claim 19 to 31, wherein said medicine also comprises guanylate cyclase stimulant or stabilizing agent.

33. method according to claim 31, wherein guanylate cyclase stimulant or stabilizing agent be not with the molecule or the ion of borine carbonate compound or ions binding.

34. claim 32 or 33 described methods, wherein guanylate cyclase stimulant or stabilizing agent are YC-1.

35., comprise with borine carbonate compound or ion and guanylate cyclase stimulant or simultaneously or in a sequence administration of stabilizing agent according to any described method in the claim 32 to 34.

36. according to any described purposes in the claim 19 to 35, wherein borine carbonate compound or ion are different from:

I. K ₂(H ₃BCOO)

Wherein R, R '=H, alkyl, perfluoroalkyl.

37. the method for the mammalian organs that an ex vivo treatment can be survived or the mammalian organs of separation comprises described organ is applicable to and provides the borine carbonate compound that CO brings into play physiological action or the pharmaceutical composition of ion to contact with containing.

38. according to the described method of claim 37, wherein borine carbonate compound or ion are defined in as claim 5 to 13 any one.

39. according to claim 38 or 39 described methods, wherein said composition also comprises guanylate cyclase stimulant or stabilizing agent.

40. according to the described method of claim 39, wherein guanylate cyclase stimulant or stabilizing agent be not with the molecule or the ion of borine carbonate compound or ions binding.

41. according to claim 39 or 40 described methods, wherein guanylate cyclase stimulant or stabilizing agent are YC-1.

42. medical treatment or veterinary transplantation body, it carries with releasable form at transplantation site borine carbonate compound or the ion that provides CO to bring into play physiological action is provided.

43. according to the described transplant of claim 38, wherein borine carbonate compound or ion are defined in as claim 5 to 13 any one.

44. according to claim 42 or 43 described transplant, wherein said medicine also comprises guanylate cyclase stimulant or stabilizing agent.

45. according to the described transplant of claim 44, wherein guanylate cyclase stimulant or stabilizing agent be not with the molecule or the ion of borine carbonate compound or ions binding.

46. according to claim 44 or 45 described transplant, wherein guanylate cyclase stimulant or stabilizing agent are YC-1.

47. one kind imports mammiferous method with CO as therapeutic agent, comprising: a) administration can provide the borine carbonate compound of the CO that is suitable for bringing into play physiological action; And b) administration guanylate cyclase stimulant or stabilizing agent.

48. method according to claim 47, this method with CO as the transmission that is used to excite nerve of physiology potent agent, vasodilation or smooth muscle relaxation, perhaps be used for the treatment of any hypertension, radiation damage, endotoxic shock, inflammation, the disease relevant with inflammation, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, organ damage behind the ischemic, miocardial infarction, angina pectoris, hemorrhagic shock, sepsis, the telotism dysfunction, ARDS, reangiostenosis, cirrhosis, cardiomegaly, heart failure and ulcerative colitis perhaps are used for the sacculus angioplasty, treatment in artery transplantation or the transplant organ survival.

49. method according to claim 47, this method with CO as the transmission that is used to excite nerve of physiology potent agent, vasodilation or smooth muscle relaxation, perhaps be used for the treatment of any acute or chronic systemic hypertension, radiation damage, endotoxic shock, the damage that hyperoxia causes, Apoptosis, cancer, graft-rejection, post operative ileus, arteriosclerosis, organ damage behind the ischemic, angina pectoris, hemorrhagic shock, sepsis, the telotism dysfunction, reangiostenosis, cirrhosis, cardiomegaly, heart failure and ulcerative colitis perhaps are used for the sacculus angioplasty, treatment in artery transplantation or the transplant organ survival.

50. according to the described method of claim 47, this method with CO as the transmission that is used to excite nerve of physiology potent agent, vasodilation or smooth muscle relaxation, perhaps be used for the treatment of any acute or chronic systemic hypertension, the damage that hyperoxia causes, promote the cancer that apoptotic effect causes by CO, graft-rejection, post operative ileus, organ damage behind the ischemic, angina pectoris, hemorrhagic shock, the penile erectile function obstacle, cirrhosis, cardiomegaly, heart failure and ulcerative colitis perhaps are used for the treatment of sacculus angioplasty or artery transplantation.

51. method according to claim 47, this method is used for the treatment of organ damage, miocardial infarction, penile erectile function obstacle, reangiostenosis, cirrhosis, cardiomegaly, heart failure, chronic anal, musculus sphincter ani internus disease, anorectal disease and the ulcerative colitis behind any acute or chronic systemic hypertension, pulmonary hypertension, graft-rejection, post operative ileus, arteriosclerosis, the ischemic, perhaps is used for the treatment of sacculus angioplasty or artery transplantation.

52. according to any described method in the claim 47 to 51, wherein borine carbonate compound or ion are defined in as claim 5 to 13 any one.

53. according to any described method in the claim 47 to 52, wherein guanylate cyclase stimulant or stabilizing agent be not with the molecule or the ion of borine carbonate compound or ions binding.

54. according to any described method in the claim 47 to 52, wherein guanylate cyclase stimulant or stabilizing agent are YC-1.

55. a pharmaceutical composition comprises:

A) can provide borine carbonate compound or the ion of the CO that is suitable for bringing into play physiological action; With

B) guanylate cyclase stimulant or stabilizing agent.

56. according to the described composition of claim 55, wherein borine carbonate compound or ion are defined in as claim 5 to 13 any one.

57. according to claim 55 or 56 described compositions, wherein guanylate cyclase stimulant or stabilizing agent be not with the molecule or the ion of borine carbonate compound or ions binding.

58. according to any described composition in the claim 55 to 57, wherein guanylate cyclase stimulant or stabilizing agent are YC-1.

59. according to any described composition in the claim 55 to 58, a kind of in the administration of administration and order when it is suitable for borine carbonate compound or ion and guanylate cyclase stimulant or stabilizing agent.